AU2018203022B2

AU2018203022B2 - 3-aryl-3-hydroxy-2-amino-propionic acid amides, 3-heteroaryl-3-hydroxy-2-amino-propionic acid amides and related compounds having analgesic and/or immunostimulant activity

Info

Publication number: AU2018203022B2
Application number: AU2018203022A
Authority: AU
Inventors: Eric Beausoleil; John E. Donello; Bertrand Leblond; Thierry Taverne
Original assignee: Allergan Inc
Current assignee: Allergan Inc
Priority date: 2005-01-26
Filing date: 2018-05-01
Publication date: 2019-07-18
Anticipated expiration: 2026-01-25
Also published as: AU2018203022A1; AU2015275339B2; AU2015275339A1

Abstract

3-aryl-3-hydroxy-2-amino-propionic acid amides, 3-heteroayryl-3-hydroxy-2-amino propionic acid amides and related compounds having analgesic and/or immunostimulant activity Abstract Compounds of Formulas (1) and (2), where the variables have the meaning disclosed in the specification, have analgesic and in some cases immunostimulant activity. OR 4 I R N Rs WV . (1) I12)

Description

3-ARYL-3-HYDROXY-2-AMINO-PROPIONIC ACID AMIDES, 3HETEROARYL-3-HYDROXY-2-AMINO-PROPIONIC ACID AMIDES AND RELATED COMPOUNDS HAVING ANALGESIC AND/OR IMMUNO STIMULANT ACTIVITY

BACKGROUND OF THE INVENTION

Claim Of Priority: The present application claims the priority of United States provisional application serial number 60/647,271 filed on January 26, 2005.

Field of the Invention

The present invention relates to derivatives of 3-aryl-3-hydroxy-2-aminopropionic acid amides, 3-heteroaryl-3-hydroxy-2-amino-propionic acid amides ' z and to related compounds having analgesic and in some cases immuno stimulant activity. . .

The present invention also relates to pharmaceutical compositions containing these compounds as active ingredient for alleviating or eliminating pain in mammals and/or stimulating the immune system in mammals and to methods of using said pharmaceutical compositions as analgesics and or immuno stimulants.

Background Art

Several compounds falling within one or more of the general definitions as “derivatives of 3-aryl-3-hydiOxy-2-amino-piOpionic acid amides, of 3heteroaryl-3-hydroxy-2-amino-piOpionic acid amides, of I-aryl-l-hydroxy-2,3diamino-propyl amines, 1-heteroaryl-l-hydroxy-2,3-diamino-propyl amines” are known in the patent and scientific literature.

For example, United States Patent Application Publications US 2003/0153768; US 2003/0050299 disclose several examples of the aboveWO 2006/081273

PCT/US2006/002557

2018203022 01 May 2018 mentioned known compounds. The TV-acyl compounds of these references are said to be useful as 7V-acylsphingosine glucosyltransferase inhibitors, the amide and the reduced compounds are described as intermediates in their preparations. Illustrative specific examples of compounds of these references are shown below:

X = H,H X = O

X = H,H X = O

The publication Shin et al. Tetrahedron Asymmetry, 2000, 11, 3293-3301 discloses the following compounds:

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2018203022 01 May 2018

(1 R,2R)-2-((S)-1 -phenylethylamino)-3morpholino-1-phenylpropan-1-ol (1R,2R)-2-amino-3-morpholino-1phenylpropan-1-ol

L-iAreo-PDMP and some other known compounds used in the methods of this invention are commercially available, in pure enantiomeric and racemic forms, as applicable, from Matreya, LLC Pleasant Gap, Pennsylvania.

United States Patent Nos. 5,945,442; 5,952,370; 6,030,995 and 6,051,598, which are all related to each other as being based on same or related disclosures, describe compounds which are structurally similar to the known compounds

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2018203022 25 Jun 2019 shown above. The compounds of these U.S. patent references are said to be inhibitors of the enzyme glucosylceramide (GlcCer) synthethase.

A publication in Journal of Labelled Compounds & Radiopharmaceuticals (1996),

38(3), 285-97 discloses the compound of the formula

Published PCT application WO 01/3 8228 discloses

OH O

in connection with a chromatographic method.

Kastron et al. in Latvijas PSR Zinatnu Akademijas Vestis, Kimijas Serija (1965) (4), 474-7 disclose the following compound.

Significantly, according to the best knowledge of the present inventors, none of the compounds of the prior art which are structurally similar to the novel compounds of the present invention are known in the prior art as analgesics or immunostimulants.

SUMMARY OF THE INVENTION

In a first aspect the present invention provides a compound represented by the following Formula 1:

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4A

Formula 1 or a pharmaceutically acceptable salt thereof, wherein:

Ri and R₂ are C1-3 alkyl, or Ri and R₂ jointly form a pyrrolidine, 3-fluoropyrrolidine, 3,3difluoropyrrolidine, 3-hydroxypyrrolidine, morpholine, thiomorpholine, piperazine, C1-8 alkylpiperazine, azetidine, tetrahydrothiazole, indoline or 2H-pyrrole group;

each R₃ is independently selected from H, Ci-₂o alkyl, C3.6 cycloalkyl, aryl-C 1.4 alkyl, aryl(hydroxy)Ci-4 alkyl, COR7 and CO2R7, the aryl groups optionally being substituted with 13 groups independently selected from halogen, C1-8 alkyl, C1-8 alkoxy and C1-3 thioxy;

R7 is H, Ci-₂o alkyl, aryl or aryl-Ci-4 alkyl, the Ci-₂o alkyl group optionally being substituted with NH₂, and the aryl groups optionally being substituted with 1 to 3 groups independently selected from halogen, C1-6 alkyl, C1-6 alkoxy and C1-3 thioxy;

R4 is H or CORs;

Rs is Ci-6 alkyl;

the wavy lines represent bonds connected to carbons having an R or S configuration; the dashed lines represent optional bonds, with the proviso that the ring containing the dashed lines is aromatic;

m, n and q are integers independently selected from 0, 1, 2 and 3, with the proviso that the sum of m, n and q is 2 or 3;

s is zero, except when X is N, in which case s is 0 or 1;

W, X and Y independently represent CH, CR5, CR6 or a heteroatom selected from N, O and S, with the proviso that when the sum of m, n and q is 2, one or two of W, X and Y is a heteroatom, and when the sum of m, n and q is 3, none or one of W, X and Y is a heteroatom; and

R5 and R6 are independently selected from H, halogen and C1-6 alkyl, or R5 and R6, together with the atoms to which they are attached, jointly form a carbocyclic or

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4B heterocyclic ring having 5 or 6 atoms in the ring, 1 to 3 atoms in the heterocyclic ring being independently selected from N, O and S;

with the proviso that Formula 1 does not cover compounds where R4 is H, Ri and R2 jointly with the nitrogen atom form a pyrrolidine or morpholine ring, the sum of m, n and q is 3, and none of W, X and Y represents a heteroatom.

In a second aspect the present invention provides a compound represented by the following Formula 2:

or a pharmaceutically acceptable salt thereof, wherein:

Ri and R2 form a saturated or unsaturated 5- or 6-membered ring together with the nitrogen atom, the ring optionally including one or two additional heteroatoms independently selected from N, O and S, the ring optionally being substituted with one or two COOH, CH2OH, OH, B(OH)2, halogen, cyano or C1-6 alkyl groups/atoms, or one or two carbon atoms of the ring being bonded to an oxygen atom to form a keto group, and the ring optionally being condensed with an aromatic or non-aromatic 5- or 6-membered ring which optionally includes one or two heteroatoms selected from N, O and S; each R3 is H, or one R3 is H and the other R3 is benzyl, monohalogen-, dihalogen-, methyl- or methoxysubstituted benzyl, cyclohexyl, C1-7 alkyl, COR7 or COOR7;

the wavy lines represent bonds connected to carbons having an R or S configuration; the dashed lines represent an optional bond, with the proviso that the ring containing the dashed lines is aromatic; m, n and q are integers independently selected from 0, 1, 2 and 3, with the proviso that the sum of m, n and q is 2 or 3; W, X and Y each independently represent CH, CR5, CR6 or a heteroatom selected from N, O and S, with the proviso that when the sum of m, n and q is 2, one or two of W, X and Y is a heteroatom, and when the sum of m, n and q is 3, none or one of W, X and Y is a heteroatom; s is 0, except when X is N, in which case s is 0 or 1;

R13 and Rio are one of the following combinations of (1) to (3): (1) R13 is NOR11 and Rio does not exist; (2) R13 is OR11 and Rio is C1-6 alkyl; and (3) when the dashed line between

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4C carbons 2 and 3 of the propionic acid moiety represents a bond then R13 is C1-6 alkyl; and

Rio does not exist;

R11 is H, C1-6 alkyl or COR12;

R12 is C1-6 alkyl;

Rs and R6 are independently selected from H, halogen, C1-6 alkyl, halogen-substituted C1-6 alkyl, C1-6 alkoxy, C1-3 thioxy and phenyl, or R5 and R6, together with the atoms to which they are attached, jointly form a carbocyclic or heterocyclic ring having 5 or 6 atoms in the ring, 1 to 3 of the atoms in the heterocyclic ring being heteroatoms independently selected from N, O and S, and the carbocyclic or heterocyclic ring being optionally substituted with to 6 R9 groups; and each R9 is independently selected from halogen, C1-6 alkyl, C1-6 alkoxy and C1-3 thioxy.

In a third aspect the present invention provides a compound represented by the following Formula 2:

or a pharmaceutically acceptable salt thereof, wherein:

Ri, R2, R3, Rs, Re, m, n, q, s, W, X, Y, the wavy lines and the dashed lines are as defined in Claim 1;

R13 is NOR11 and Rio does not exist, or R13 is OR11 and Rio is C1-6 alkyl;

R11 is H or COR12; and

R12 is Ci-6 alkyl.

In a fourth aspect the present invention provides a pharmaceutical composition comprising a compound as claimed in the first, second or third aspects, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients.

In a fifth aspect the present invention provides use of a compound as claimed in the first, second or third aspects, a pharmaceutically acceptable salt thereof, or a composition according to the fourth aspect, in the manufacture of a medicament for the treatment of pain in a mammal.

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4D

In a sixth aspect the present invention provides use of a compound as claimed in the first, second or third aspects, a pharmaceutically acceptable salt thereof, or a composition according to the fourth aspect, in the manufacture of a medicament for stimulating the immune system of a mammal.

In a seventh aspect the present invention provides use of a compound of the formula:

or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treatment of chronic pain associated with peripheral neuropathy in a mammal, wherein: Ri and R2 are each independently H or C1-6 alkyl, or Ri and R2 together with the nitrogen atom to which they are attached form a saturated or unsaturated 4, 5, 6, 7 membered ring, the ring optionally including one or two additionally heteroatoms independently selected from N, O and the ring optionally being substituted with one or two CO2H, CH2OH, OH, B(OH)2, halogen, cyano or C1-6 alkyl groups;

each R3 is independently H, COR7 or CO2R7;

R7 is H, C1-20 alkyl, C3-6 cycloalkyl, aryl, heteroaryl, aryl-Ci-4 alkyl, aryl-(hydroxy)Ci-4 alkyl, heteroaryl-Ci-4 alkyl or heteroaryl-(hydroxy)Ci-4 alkyl, the aryl and heteroaryl groups being optionally substituted with 1 to 3 groups independently selected from halogen, C1-6 alkyl, C1-6 alkoxy and C1-3 thioxy;

R4 is H, C1-6 alkyl or CORs;

Rs is Ci-6 alkyl;

R5 and R6 are independently selected from H, halogen, C1-6 alkyl, C1-6 alkoxy and C1-3 thioxy, or Rs and R6 together with the atoms to which they are attached jointly form a carbocyclic or heterocyclic ring having 5 or 6 atoms in the ring, the heterocyclic ring having 1 to 3 heteroatoms independently selected from N, O and S, and the carbocyclic and heterocyclic rings being optionally substituted with 1 to 6 R9 groups; each R9 is independently selected from halogen, C1-6 alkyl and C1-6 alkoxy; and the wavy lines represent bonds of the alpha or beta configuration.

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4E

In an eighth aspect the present invention provides a method for treating pain in a mammal in need thereof, the method comprising administering to the mammal a compound as claimed in the first, second or third aspects, or a pharmaceutically acceptable salt thereof, or a composition as claimed in the fourth aspect.

In a ninth aspect the present invention provides a method for stimulating the immune system of a mammal in need thereof, the method comprising administering to the mammal a compound as claimed in the first, second or third aspects, or a pharmaceutically acceptable salt thereof, or a composition as claimed in the fourth aspect.

In a tenth aspect the present invention provides a method for treating chronic pain associated with peripheral neuropathy in a mammal in need thereof, the method comprising administering to the mammal a compound as defined in the seventh aspect, or a pharmaceutically acceptable salt thereof.

The present invention is directed to compounds of Formula 1

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where Rj is H or alkyl of 1 to 6 carbons,

R₂ is H, alkyl of 1 to 6 carbons or the Ri and R₂ groups together with the nitrogen form a saturated or unsaturated 4, 5, 6 or 7 membered ring that optionally includes one or two heteroatoms independently selected from N, O and S, said 4, 5, 6 or 7 membered ring optionally being substituted with one or two COOH, CH₂OH, OH, B(OH)₂, cyano or halogen groups or with one or two alkyl groups having 1 to 6 carbons, or one or two carbons of said rings being attached to an oxygen to form keto groups and said 4, 5, 6 or 7 membered ring optionally being condensed with an aromatic or noil-aromatic 5 or 6 membered ring that optionally includes 1 or heteroatoms selected fromN, O and S;

R₃ is independently selected from H, alkyl of 1 to 20 carbons, cycloalkyl of 3 to 6 carbons, aryl or heteroaryl, aryl-alkyl, aryl-(hydroxy)alkyl, heteroaryl-alkyl or hetero-(hydroxy)alkyl where the alkyl moiety has 1 to 4. carbons, said aryl or heteroaryl groups being optionally substituted with 1 to 3 groups independently selected from the group consisting of halogen, alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to 6 carbons, or R₃ is CO-R7, SO₂R7 or CO-O-R7 where R₇ is H, alkyl of 1 to 1 to 20 carbons, alkyl of 1 to 20 carbons substituted with and NH₂ group or with an NH-COalkyl group where the alkyl group has one to 6 carbons, aryl or heteroaryl, aryl-alkyl or heteroaryl-alkyl where the alkyl moiety has 1 to 4 carbons, said aryl or heteroaryl groups being optionally substituted with 1 to 3 groups independently selected from the group consisting

WO 2006/081273

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2018203022 01 May 2018 of halogen, alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to carbons;

R4 is H, alkyl of 1 to 6 carbons or CO-R₈ where R₈ is alkyl of 1 to 6 carbons;

the wavy lines represent bonds connected to carbons having R or 5 configuration;

the dashed lines represent a bond or absence of a bond with the proviso that the ring containing the dashed lines is aromatic;

m, 11 and q are integers independently selected from 0, 1, 2 or 3 with the proviso that the sum of 111,11 and q is 2 or 3;

s is zero (0) or when X is N then s is zero (0) or 1;

W, X and Y independently represent a CH, CR5, CR6 or a heteroatom selected independently of N, O and S, and

R₅ and R₆ are independently selected from H, halogen, alkyl of 1 to 6 carbons, halogen substituted alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to 6 carbons, phenyl, or

R₅ and R₆ together with the atoms to which they are attached jointly form a carbocyclic or a heterocyclic ring, the carbocyclic ring having 5 or 6 atoms in the ring, the heterocyclic ring having 5 or 6 atoms in the ring and 1 to 3 heteroatoms independently selected fromN, O and S;

said carbocyclic or heterocyclic ring jointly formed by R₅ and R₆ being . optionally substituted with 1 to 6 R₉ groups where R₉ is independently selected from halogen, alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to 6 carbons or a pharmaceutically acceptable salt of said compound with the proviso that Formula 1 does not cover compounds where R4 is H, Rj and R₂ jointly with the nitrogen form a pyrrolidino or morpholino ring, the sum of m, 11 and q is 3, and none of W, X and Y represent a heteroatom with the

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2018203022 25 Jun 2019

nh₂

OH 0

NH₂

ΝΗ_ξ

DL-eryfftro

The present invention is also directed to the compounds of Formula 2

where Ri is H or alkyl of 1 to 6 carbons,

R2 is H, alkyl of 1 to 6 carbons or the Ri and R2 groups together with the nitrogen form a saturated or unsaturated 4, 5, 6 or 7 membered ring that optionally includes one or two heteroatoms independently selected from N, O and S, said 4, 5, 6 or 7 membered ring optionally being substituted with one or two COOH, CH2OH, OH, B(OH)2, cyano or halogen groups or with one or two alkyl groups having 1 to 6 carbons, or one or two carbons of said rings being attached to an oxygen to form keto groups and said 4, 5, 6 or 7 membered ring optionally being condensed with an aromatic or non-aromatic 5 or 6 membered ring that optionally includes 1 or heteroatoms selected from N, O and S;

R3 is independently selected from H, alkyl of 1 to 20 carbons, cycloalkyl of 3 to 6 carbons, aryl or heteroaryl, aryl-alkyl, aryl-(hydroxy)alkyl, heteroaryl-alkyl or hetero(hydroxy)alkyl where the alkyl moiety has 1 to 4 carbons, said aryl or heteroaryl groups being optionally substituted with 1 to 3 groups independently selected from the group consisting of halogen, alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to 6 carbons, or R3 is CO-R7, SO2R7 or CO-O-R?where R7 is H, alkyl of 1 to 1 to 20 carbons, alkyl of 1 to 20 carbons substituted with an NH2, NHCOR7 or NHCOOR7 group, aryl or heteroaryl, aryl-alkyl or heteroaryl-alkyl where the alkyl moiety has 1 to 4 carbons, said aryl or heteroaryl groups being optionally substituted with 1 to 3 groups independently selected from the group consisting of halogen, alkyl of 1 to 6 carbons, alkoxy of 1 to 6

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2018203022 25 Jun 2019 carbons and thioxy of 1 to 6 carbons;

the wavy lines represent bonds connected to carbons having R or S configuration;

R13 and Rio are independently H, alkyl of 1 to 6 carbons or ORh, or R13 and Rio jointly represent NOR11 with the proviso that when the dashed lines between carbons 2 and 3 of the propionic acid moiety represents a bond then Rio does not exist and R13 is not OR11 with the further proviso that when R13 is OR11 then Rio is not hydrogen;

R11 is H, alkyl of 1 to 6 carbons or CO-R12 where R12 is alkyl of 1 to 6 carbons;

m, n and q are integers independently selected from 0, 1, 2 or 3 with the proviso that the sum of m, n and q is 2 or 3;

s is zero (0) or when X is N then s is zero (0) or 1;

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said carbocyclic or heterocyclic ring jointly formed by R₅ and R₆ being optionally substituted with 1 to 6 R₉ groups where R₉ is independently selected from halogen, alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to 6 carbons or a pharmaceutically acceptable salt of said compound.

The present invention is also directed to pharmaceutical compositions containing the above-noted novel compound to be used as analgesics and/or immunostimulants in mammals, and to methods of using said pharmaceutical compositions as analgesics or immunostimulants.

DETAILED DESCRIPTION OF THE INVENTION

A general description of the compounds of the invention is provided in the Summary Section of the present application for patent. Most compounds of the invention contain one or more asymmetric centers, such that the compounds may exist in enantiomeric as well as in diastereomeric forms. In fact, most of the compounds of the present invention have two asymmetric carbons adjacent to one another and therefore can exist in erythro or threo form, with each of these two forms having dextrorotatory (D) or levorotary (L) enantiomers. Although the threo form is generally preferred in accordance with the present invention for analgesic activity, unless it is specifically noted otherwise, the scope of the present invention includes all enantiomers, diastereomers and diastereomeric or racemic mixtures. In light of the foregoing, it should be

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2018203022 01 May 2018 clearly understood that the designation “DL” or “(+/-)” or “(±)” in this application includes the pure dextrorotatory enantiomer, the pure levorotatory enantiomer and all racemic mixtures, including mixtures where the two enantiomers are present in equal or in unequal proportions. Moreover, for simplicity sake in many of the structural formulas, such as in the example below, only one of the enantiomers is actually shown but when the designation “DL” or “(+/-)” or or “(±)” appears it also includes the enantiomeric form (mirror image) of the structure actually shown in the formula.

For Example:

OL-threo (only one enantiomer shown)

Thus, in the example above, only one enantiomer is shown, but because the designation “DL” (or or “(+/-)” or “(±)”) appears below the formula, its optical isomer oh o

HCI

DL-threo (the other enantiomer shown) and all racemic mixtures of the two optical isomers are also included.

In the case of some compounds of the present invention one enantiomer of the threo, and in some cases of the erythro, enantiomers is significantly more active as an analgesic than the other enantiomer of the same pair. For this, reason the isolated enantiomer which is significantly more active than the other is

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2018203022 01 May 2018 considered a novel and inventive composition even if the racemic mixture or the other opposite enantiomer of the same compound have already been described in the prior art.

Some of the novel compounds of the present invention contain three or more asymmetric centers. An example is the following compound oh o

2.HCI (2S,3R) & (2R,3S)

Compound 214 named Compound 214 in the description. The formula shown in the description for Compound 214 indicates two compounds of the threo isomer, but the two compounds indicated are not mirror images of each other, they are diastereomers. Another isomer pair is shown and described as Compound 215.

oh 0

N . i > F

NH₂ ^/(^r>

2.HCI (2S,3R) & (2R,3S)

Compound 215

Keeping the foregoing examples in mind the reader one of ordinary skill in the art should readily understand the scope of each described example, although in a broad sense all isomers, enantiomers and racemic mixtures are within the scope of the invention.

The term “alkyl” in the general description and definition of the compounds includes straight chain as well as branch-chained alkyl groups.

Generally speaking the compounds of the invention may form salts with pharmaceutically acceptable acids or bases, and such pharmaceutically

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2018203022 01 May 2018 acceptable salts of the compounds of Formula 1 and of Formula 2 are also within the scope of the invention.

Referring now to the novel compounds of Formula 1, in a class of preferred compounds of the invention none of the W, X and Y groups is a heteroatom. Within this class, compounds are preferred where the sum of m, n and q is 3 and the aromatic group is unsubstituted or substituted with one or more halogen, alkyl of 1 to 6 carbons, or halogen substituted alkyl of 1 to 6 carbons. Compounds within this class are also preferred where the R₅ and R₆groups form a carbocyclic ring, or a heterocyclic ring.

In another class of preferred compounds in accordance with Formula 1 one of the variables W, X and Y represents a heteroatom, preferably nitrogen and the sum of m, n and q is 3.

In still another class of preferred compounds in accordance with Formula 1 one or two of the variables W, X and Y represent a heteroatom, selected from N, O or S and the sum of m, u and q is 2.

Referring still to the compounds of Formula 1, compounds are preferred where R₄ is H or an acyl group, more preferably H.

With reference to the variables R₃, compounds in accordance with Formula 1 are preferred where both R₃ groups are Hand where one R₃ group is H and the other is benzyl, monohalogeno, dihalogeno, methyl or methoxy substituted benzyl, cyclohexyl, an alkyl of 1 to 7 carbons, COR₇, COOR7 where . R₇ is alkyl of 1 to 15 carbons, benzyloxy, phenyl, methoxyphenyl, monohalogen or dihalogeno substituted phenyl, a 2-hydroxy-l -phenylethyl group or an alkyl group of 1 to 20 carbons itself substituted with an NH₂, NHCOR₇, or NHCOOR₇ group.

Referring now to the variables Ri and R₂ in the compounds of Formula 1, compounds are preferred in accordance with the invention where Ri and R?

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2018203022 01 May 2018 jointly form a pyrrolidine, a 3-fluoro or a 3,3-difluoro or an 3-hydroxy substituted pyrrolidine, a morpholine, a thiomorpholine, a piperazine, an alkyl substituted piperazine where the alkyl group has 1 to 6 carbons, an azetidine, a tetrahydrothiazole, an indoline, or a 277-pyrrol ring or Ri and IL are two alkyl groups of 1 to 3 carbons.

Referring now to the novel compounds of Formula 2, with respect to the variables W, X, Y, m, n, q, R_b R₂, R₅, R₆, R₃ compounds are gerenerally preferred in which these variables have the same preferences as in compounds of Formula 1.

With respect to R₉ and Ri₀, compounds are generally preferred where R₉and Rio are both hydrogen, where one of these two variables is hydroxy and the other is alkyl of 1 to 6 carbons, where the R₉ and R₁₀ groups jointly form an NORn group, and where R₉ is hydrogen, the dashed line between carbons 2 and 3 represent a double bond and Rio does not exist. With respect to Rn compounds of Formula 2 are preferred where Rn is H, or CORi₂ where R_i2 is alkyl of 1 to 3 carbons.

Presently still more preferred are Compounds of Formula 2 where Ri and R₂ jointly with the nitrogen form a five-membered ring, where both R₃ groups are hydrogen and where one of the R₃ groups is hydrogen and the other is formyl.

The presently most preferred novel compounds of the invention are disclosed with their structural formulas in the ensuing Tables and or description, . showing activity of exemplary compounds relevant to their ability to act as analgesics and/or immunostimulants.

BIOLOGICAL ACTIVITY, MODES OF ADMINISTRATION

The novel compounds of the invention have analgesic and/or immunostimulant activity in mammals. Some of the compounds described in

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2018203022 01 May 2018 the introductory section which per se are known in the art, have been discovered by the present inventors to also have analgesic effect in mammals. To the best of the knowledge of the present inventors the analgesic or immunostimulant biological activity of the known compounds was not known before the present discovery.

An art-accepted model or assay for measuring an analgesic effect of a compound in chronic pain (in particular peripheral neuropathy) is the model known as Kim and Chung 1992, Pain 150, pp 355-363 (Chung model). This model involves the surgical ligation of the L5 (and optionally the L6) spinal nerves on one side in experimental animals,. Rats recovering from the surgery gain weight and display a level of general activity similar to that of normal rats. However, these rats develop abnormalities of the foot, wherein the hindpaw is moderately everted and the toes are held together. More importantly, the hindpaw on the side affected by the surgery appears to become sensitive to lowthreshold mechanical stimuli and will perceive pain instead of the faint sensation of touch. This sensitivity to normally noil-painful touch, called “tactile allodynia”, develops within the first week after surgery and lasts for at least two months. The allodynia response includes lifting the affected hindpaw to escape from the stimulus, licking the paw and holding it in the air for many seconds. None of these responses is normally seen in the control group.

To produce the tactile allodynia, rats are anesthetized before surgery. The surgical site is shaved and prepared either with betadine or Novacaine. Incision is made from the thoracic vertebra Xlll down toward the sacrum. Muscle tissue is separated from the spinal vertebra (left side) at the L4 - S2 levels. The L6 vertebra is located and the transverse process is carefully removed with a small rongeur to expose the L4 - L6 spinal nerves. The L5 and L6 spinal nerves are isolated and tightly ligated with 6-0 silk thread. The same

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2018203022 01 May 2018 procedure is done on the right side as a control, except no ligation of the spinal nerves is performed.

After a complete hemostasis is confirmed, the wounds are sutured. A small amount of antibiotic ointment is applied to the incised area, and the rat is transferred to the recovery plastic cage under a regulated heat-temperature lamp.

On the day of the experiment, at least seven days after the surgery, typically six rats per test group are administered the test drugs by intraperitoneal (i.p.) injection or oral gavage (p.o.). For i.p. administration, the compounds are formulated in H₂O and given in a volume of 1 ml/kg body weight by injecting into the intraperitoneal cavity. For p.o. administration, the compounds are formulated in H₂O and given in a volume of 1 ml/kg body weight using an 18gauge, 3 inch gavage needle that is slowly inserted through the esophagus into the stomach.

Tactile allodynia is’assessed via von Frey hairs, which are a series of fine hairs with incremental differences in stiffness. Rats are placed in a plastic cage with a wire mesh bottom and allowed to acclimate for approximately 30 minutes. To establish the pre-drug baseline, the von Frey hairs are applied perpendicularly through the mesh to the mid-plantar region of the rats’ hindpaw with sufficient force to cause slight buckling and held for 6-8 seconds. The applied force has been calculated to range from 0.41 to 15.1 grams. If the paw is sharply withdrawn, it is considered a positive response. A normal animal will not respond to stimuli in this range, but a surgically ligated paw will be withdrawn in response to a 1-2 gram hair. The 50% paw withdrawal threshold is determined using the method of Dixon, W.J., Ann. Rev. Pharmacol. Toxicol. 20:441-462 (1980) hereby incorporated by reference. Tactile allodynia is measured prior to and 15, 30, and 60 minutes after drug administration. The post-drag threshold is compared to the pre-drug threshold and the percent

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2018203022 01 May 2018 . reversal of tactile sensitivity is calculated based on a normal threshold of 15.1 grams.

Table 1 below indicates the degree of pain reversal obtained in the Chung model with exemplary compounds of the invention. The intraperitonial (i.p.) and/or intravenous (iv) administration of the compounds was in doses ranging from 1 pg/kg to 300 pg/kg or 3 mg/kg PO and the peak percentage of reversal of allodynia was measured at 15, 30 or 60 minutes after administration, as is indicated in the table. Data are expressed as the highest % allodynia reversal (out of 3 time points: 15 min, 30 min, or 60 min. post-ding) with a minimum of a 20% allodynia reversal in the rat Chung model. Comparisons between groups (drug treated vs. saline treated) were made using a two-tailed, 2-sample, unpaired t-test. Compounds that are not shown which were not statistically analgesic following an IP dose of 300 ug/kg, but may still be analgesic. Compounds that do not exhibit significant analgesia at 100 mg/kg are not considered to be analgesic.

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TABLE 1

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22	OH 0 NH₂ '—V 2.HCI DL-f/ireo	100% ' 60 min	3000μ g/kg PO
22	OH O NH₂ U-V 2.HCI DL-threo	100% 60 min	100 gg/kg IP
20	OH 0 Ah₂ '—' 2.HCI DL-threo	92% 60 min	30 μg/kg IP
35	OH 0 foH₂/ 2.HCI DL-threo	92% 60 min	300 μg/kg IP
23	OH O A \jj nh₂ W HCI DL-threo	100% 60 min	30 μg/kg PO
24	OH O O HCI DL-threo	60% 60 min	300 μg/kg IP

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58	OH 0 hci DL-threo —'J	75% 60 min	300 fig/kg IP
59	OH 0 A HN '—/ HCI DL-threo \	92% 60 min	300 qg/kg IP
27	OH 0 Y nh₂ W HCI DL-t/ireo	42% 30 min	300 fig/kg ip
29	i/X OH 0 TfAx YJI Ah₂ HCI DL-f/ireo	47% 60 min	300 fig/kg IP
61	OH 0 ΐΥϊ^Τ^Ν-'Χ nY HFl '—/ 2 HCI ^X DL-threo	64% 60 min	300 fig/kg IP
34	OH 0 NH₂ U-# 2.HCI DL-f/?reo	62% 60 min	300 gg/kg IP

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Compound #	Chemical Formula	Peak % Pain reversal: time post dose	Dose pg/kg, Mode of administ.
30	OH 0 Υ/γΑθ Yv ^NH2 2.HCI DL-threo	53% 60 min	300 pg/kg IP
64	OH O Y hr Ύ 2.HCI DL-threo \	100% 30 min	300 pg/kg IP
55	OH O nY NH L—/ HCI DL-threo	58% 60 min	300 pg/kg IP
56	OH O NY NH ¹—/ HCI DL-threo \Y₀ 1	67% 60 min	300 pg/kg IP
67	OH O nY NH L—/ 2.HCI DL-threo Y_Q/	78% 60 min	300 pg/kg IP

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Compound #	Chemical Formula	Peak % Pain reversal: time post dose	Dose Pg/kg, Mode of administ.
68	OH 0 ίγΑο NH L-J 2.HCI OL-threo Η Ϊ ^k>ci	94% 60 min	300 pg/kg PO
69	OH O [fVYi'A N^9 NH ¹—/ 2.HCI DL-t/ireo	63% 30 min	300 pg/kg IP .
41	OH O r> ⁷ B kL..J· nh₂ * 2.HCI QL-tlireo '	70% 60 min	300 pg/kg IP
49	O OH O H 7 Π ΗίΤΥγΧΛ ' k^¹-! ^H₂ HCI ” ’ QL-threo	85% 60 min	100 pg/kg IP
43	Cl OH O ftH₂ W . 2.HCI DL.-threo	96% 60 min	300 pg/kg IP

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Compound #	Chemical Formula	Peak % Pain reversal: time post dose	Dose Pg/kg, Mode of administ.
26	OH 0 g·—' NH₂ '—< HCI DL-threo	92% 60 min	300 pg/kg / IP
57	OH O NH CV hci oApyd DL-threo	51% 30 min	300 pg/kg IP
28	OH O nh₂ cy HCI (+/-)	92% 60 min	300 pg/kg IP

Compound 216	OH O HN. '—' 11 O HCI (+)-f/?reo	100% 60 min	300 pg/kg IP
Compound 234	OH O HA ²HCI Q	59% 60 min	300 pg/kg IP
Compound 230	OH 0 γΑΛό V nh₂ HCI (±)-f/?reo	52% 30 min	300 pg/kg IP

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Compound 236	0 onto 2HCI	32% 60 min	300 pg/kg IP
Compound 218	OH o ^N-O N^JJ HN_v O/ ^HCI °v- (±)-i/ireo /\	32% 60 min	300 pg/kg IP
Compound 239	„ ^{0H 0} N^¹ NH₂ CV 2.HCI (±)-erythro	75% 30 min	300 pg/kg IP
Compound 238	OH O ^HNo^z '—\ (±)-threo ^_Nh₂	61% 60 min	300 pg/kg IP
Compound 205	OH O HCI (+)-threo	62% 30 min	300 pg/kg IP
Compound 206	OH O C0to HCI (-)-threo	67% 30 min	300 pg/kg IP
Compound 240 ,	O nh₂ Ly 2 HCI	70% 60 min	30 pg/kg IP
Compound 232	OH O N^zVy^xN^Zx\| FlH₂ 2.HCI (±)-threo	80% 60 min	300 pg/kg IP

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Compound 220

! , . OH . O. . ,. ή / NH_2! A/> • ·· 2.HCI ' 'i ' (-)-erythro ,

32% 30 min

300 pg/kg IP

Compound 210	OH O ΓγΧο XA nh₂ / HCI (±)-f/ireo	78% 60 min	300 pg/kg IP
Compound 221	OH O nh₂ A/ 2.HCI • (+)-^erythro	87% 60 min	300 pg/kg IP
Compound 227	OH O N^l Ah L—/ 2.HCI. (±)-threo 1	95% 30 min	300 pg/kg IP
Compound 226	OH O n-A NH A/ 2.HCI (+)-threo \|/\^	95% 60 min	300 pg/kg IP
Compound 207	OH O nh₂ A/ 2.HCI (-)-threo	96% 60 min	300 pg/kg IP

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Compound #	Chemical Formula	Peak % Pain reversal: time post dose	Dose pg/kg, Mode of administ.
Compound 213	OH 0 nh₂ M ^r 2.HCI (+}-threo	85% 30 min	300 pg/kg IP
Compound 214	OH O nh₂ 2.HCI (2S.3R) & (2R.3S)	86% 60 min	30 pg/kg IP
Compound 228	OH O ryM'o HNL NJ ^2ΗΟΙ·ά,	36% 60 min	300 pg/kg IP
Compound 229	OH O hiL '—/ ^2HCI ά,	53% 60 min	300 pg/kg IP
Compound 224	NOHO NH₂ \-~J 2.HCI I (±)	51% 60 min	300 pg/kg IP

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Compound #	Chemical Formula	Peak % Pain reversal: time post dose	Dose pg/kg, Mode of administ.
Compound 215	OH 0 N , 11F NH₂ 2.HCI (2S.3R) & (2R,3S)	73% 60 min	300 pg/kg IP
Compound 219	OH O N^V NH₂ '-S 2.HCI (+)-erythro	82% ' 60 min	30 pg/kg IP
Compound 203	QH o N<^JJ NH₂ V/ 2 HCI (-)-threo	87% 60 min	300 pg/kg IP
Compound 204	OH o NH₂ W 2 HCI (+)-f/ireo	50% 60 min	300 pg/kg IP
Compound 40	OH 0 I'V NH₂ '— 2.HCI DL-Wireo	47% 60 min	300 pg/kg IP
Compound 247	OH O nV Rh₂ C-/^fRJ 2.HCI + OH O nV nh₂ C/w 2.HCI	42% 60 min	300 pg/kg IP

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Compound #	Chemical Formula	Peak % Pain reversal: time post dose	Dose pg/kg, Mode of administ.
Compound 254	y OH O V-N NH₂ '—f 2.HCI . DL-threo	62% 60 min	300 pg/kg IP.
Compound 248	OH O nh₂ u~y<^s) ' 2.HCI (-)-threo	59% 60 min	30 pg/kg IP
Compdund 255	OH o . /ih₂ 2.HCI • + OH O kz νη₂ cy<^s> 2.HCI	60% 60 min	300 pg/kg IP

Compound

256

62% min

An art accepted method for measuring immuno stimulation comprises systemic administration of compounds to assay for the ability to stimulate the immune system, possibly due to nonspecific upregulation of the

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2018203022 01 May 2018 hemolymphoreticular system. This upregulation could result in increased numbers of lymphocytes of both T- and B-cell lineage. Although applicant does not wish to be bound by the biological theory of the immunostimulation, actual immunostimulatory efficacy of the compounds can be demonstrated in vivo by assaying splenic size in response to adminstration of the test compound to laboratory test species rats. Animals dosed at 200 mg/kg of Compound 22 of this invention exhibited a twentyfive percent increase in spleen size which demonstrates immunostimulatory potential of the compound. Generally speaking any compound that exhibits splenic enlargement following dosing of 200 mg/kg or less may be considered an immunostimulant.

Modes of Administration:

The compounds of the invention may be administered at pharmaceutically effective dosages. Such dosages are normally the minimum dose necessary to achieve the desired therapeutic effect; in the treatment of chromic pain, this amount would be roughly that necessary to reduce the discomfort caused by the pain to tolerable levels. For human adults such doses generally will be in the range 0.1-5000 mg/day; more preferably in the range 1 to 3000 mg/day, still more preferably in the range of 10 mg to 1000 mg/day. However, the actual amount of the compound to be administered in any given case will be determined by a physician taking into account the relevant circumstances, such as the severity of the pain, the age and weight of the patient, the patient’s general physical condition, the cause of the pain, and the route of administration.

The compounds are useful in the treatment of pain in a mammal; particularly a human being. Preferably, the patient will be given the compound orally in any acceptable form, such as a tablet, liquid, capsule, powder and the like. However, other routes may be desirable or necessary, particularly if the

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2018203022 01 May 2018 patient suffers from nausea. Such other routes may include, without exception, transdermal, intraperitonial, parenteral, subcutaneous, intranasal, intrathecal, intramuscular, intravenous and intrarectal modes of delivery. Another aspect of the invention is drawn to therapeutic compositions comprising the novel compounds of the invention and pharmaceutically acceptable salts of these compounds and a pharmaceutically acceptable excipient. Such an excipient may be a carrier or a diluent; this is usually mixed with the active compound, or permitted to dilute or enclose the active compound. If a diluent, the earner may be solid, semi-solid, or liquid material that acts as an excipient or vehicle for the active compound. The formulations may also include wetting agents, emulsifying agents, preserving agents, sweetening agents, and/or flavoring agents. If used as in an ophthalmic or infusion format, the formulation will usually contain one or more salt to influence the osmotic pressure of the formulation.

In another aspect, the invention is directed to methods for the treatment of pain, particularly chronic pain, through the administration of one or more of the novel or otherwise known compounds of the invention, or of pharmaceutically acceptable salts thereof to a mammal in need thereof. As indicated above, the compound will usually be formulated in a form consistent with the desired mode of delivery. '

Compounds of the invention which are immunostimulants are administered subject to the same basic principles as the compounds having analgesic activity, in doses which are best determined on a case-by-case and/or species-by-species and, in case of humans, at times on a patient-by-patient . basis. Generally speaking the effective dose will be in the range of 10 pg/kg to 200 mg/kg.

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In this regard it is noted that the compounds of the threo configuration are more likely to have the analgesic activity, compounds of the erythro configuration are more likely to have immunostimulant activity, and among the erythro compounds those with an S configuration at carbon 2 of the propionic acid moiety are likely to have stronger immiino stimulant activity.

SYNTHETIC METHODS FOR OBTAINING THE COMPOUNDS OF THE INVENTION, EXPERIMENTAL

The compound of the invention can be synthesized by utilizing the synthetic methods described in the experimental below, or such modifications of the below described experimental methods which will become readily apparent to those skilled in the art in light of the present disclosure.

GENERAL

4ΐ NMR spectra were recorded at ambient temperature with an Avance 300 (Broker) spectrometer. The compounds were analyzed by reverse phase high performance liquid chromatography (HPLC) using a Waters Autopurification System equipped with a Waters 2525 Pump, a Waters 2696 photodiode array detector, and a XTerra column (Part, No. 186000482, 5 μιη, C18, 4.5 x 50 mm).

The HPLC method used was a gradient of 5 % solvent B to 100 % in 7 min. Solvent A was H₂O with 0.05 % TFA and solvent B was CH₃CN with 0.05 % TFA (Method A).

Melting points were measured with a Biichi B-545 melting point apparatus and were uncorrected. To isolate reaction products the solvent were removed by evaporation using a vacuum rotatory evaporator, the water bath temperature not exceeding 40 °C.

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Absolute configuration of compounds of the invention, where applicable, can generally speaking be determined in accordance with methods known in the state of the art, such as X-ray christallography. Compounds 203 and 204 are mentioned as examples for which the absolute configurations were determined by X-ray christallography analysis of the corresponding (l<S)-camphanylamide D-(10) camphorsulfonic acid salt. As a result Compound 204 was assigned (2S, 3R). Its enantiomer, Compound 203 was assigned by default the (27?, 35) absolute configuration.

GENERAL SYNTHETIC ROUTES

The compound of the invention can be synthesized by utilizing the ' synthetic methods described in a general sense immediately below and in more detail in the experimental section of the present application, of by such modifications of the below described experimental methods which will become readily apparent to those skilled in the art in light of the present disclosure.

A general synthetic route to the novel compounds of the invention which are amides of substituted (+/-)-/Areo-3-hydroxy-2-aminopropionic acid of the Generalized Structure 1 is described below.

General Structure 1

In General Structure 1, for the sake of simplicity of description R* substantially corresponds to the 5, 6, or 7 membered ring structure on the left side of Formula 1 (as the formula is depicted in the Summary and in the

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C=N--CH₂ © \	4-	R** / UN	i® III (DO J	-ch₂	R** /
\		γίιν \ R**		X—	—N
r\			4/	\ R**

metliylisocyanoacetate		amine	2-Isocyano-l-(subst.-ammo)ethanone

R* —CHO strong base aldehyde (KOH) qh ⁰

nh₂ r** (+/-)-threo

General Structure 1 (major isomer) acid (HCI)

tra/is-oxazoline (major isomer

General Reaction Scheme 1

Thus, in accordance with General Scheme 1, methyl isocyanoacetate (or ethyl isocyanoacetate available commercially) is reacted with an “amine” which includes the R** groups to provide the 2-isocyanoacetic acid amide derivative shown in the general scheme. Typical examples for the amines used in the reaction are pyrrolidine, piperidine, azetidine, morpholine, 2,5-dihydro-lHpyrrole, dialkylamines such as diethylamine, 3-fluoro-, 3,3-difluoro or 3hydroxy substituted pyrrolidines. Specific examples of these “amines” abound in the experimental description. The 2-isocyanoacetic acid amide derivative is then reacted in the presence of base (such as KOH) with an “aldehyde” which includes the R* group to provide a trans “oxazoline” with high

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2018203022 01 May 2018 diastereoselectivity (trans’.cis ratios generally > 97:3) as shown in the general reaction scheme 1. The trans oxazoline is then treated with a strong acid, such as HCI, to open the ring and to provide the //zzw-3-substituted-3-hydroxy-2amino-propionic acid amides (with threo'.erythro ratios generally > 97:3) of the , invention as shown in General Scheme 1.

Compounds of Formula 1 where the amino group of formula NH(R**)₂is a weaker nucleophile, such as indoline, thiomorpholine and the like, can be made as illustrated in Reaction Scheme 2 for the synthesis of (ft)-threo-2 amino-3-hydroxy-1 -(indolin-1 -yl)-3-(pyridin-4-yl)propaii-1 -one dihydrochloride Compound 243 and (±)-//zreo-2-amino-3-hydroxy-l(thiazolidin-3-yl)-3-(pyridin-4-yl)propan-1 -one dihydrochloride Compound 242.

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(±) erythro 2 % yield

(±) threo

Compound 243 (±) threo 30 % yield

(±) threo

Compound 242

a) KOH, MeOH; b) Indoline, EDCI, TEA, HOBT, CH₂CI₂. c) HCI (1M) in MeOH cl) i. Silica Gel Chromatography.ii HCI (0.1 M) in /-PrOH e) BOC₂O, NaOH, Dioxane, f) Thiazolidine, EDCI, TEA, HOBT, CH₂CI₂; g) Silica Gel Chromatography g) HCI (1M) in MeOH.

Reaction Scheme 2

In Reaction Scheme 2 EDCI stands for l-(3-dimethylaminopropyl)ethylcarbodiimide hydrochloride; HOBT stands for 1-hydroxybenzotriazole; BOC₂O stands for di-i-butyl-dicarbonate and TEA stands for triethylamine.

As it will be readily understood by those skilled in the art, for a more general synthetic route, such as the one shown in Reaction Scheme 2, the 4WO 2006/081273

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2018203022 01 May 2018 pyridyl group can be substituted with an R* group (as defined in connection with Scheme 1) and the indoline can be susbstituted with other weak nucleophilic amines of the formula NH(R**)₂ (R** defined as in connection with Reaction Scheme 1) to provide other compounds of Formula 1 analogous to compounds 242 and 243.

Isomerically pure and/or enantiomerically pure compounds and further derivatives of the 3-substituted-34iydroxy-2-amiiio-propionic acid amides are obtained by separation techniques and reactions which, per se, are well known to the synthetic chemist. The experimental section of the present invention abounds in examples of such separation techniques and reactions. Some of the typical separation techniques and reactions are generally described below.

Separation of threo and erythro isomers, when both are formed in the reactions leading to the compounds of the invention, can typically be separated by chromatographic methods. The more abundantly formed threo isomers can also be converted into the erythro isomers by oxidizing to the ketone level the hydroxyl group in the 3 position of the propanoic acid moiety and subsequently reducing the resulting ketone to the hydroxyl level. (See, for example, the preparation of Compound 219).

Separation of enantiomeric mixtures can be performed on Chiralpack columns which are well known in the art. (See, for example, the preparation of Compound 204).

The amino function in the 2-position of the propanoic acid moiety is, generally speaking, more reactive towards acylation and carbamoylation than the hydroxyl group in the 3 position. Therefore, acylated derivatives of the 2amino function can be prepared by using acyl chlorides such as acetyl chloride and hexanoyl chloride. (See, Method G and the preparation of Compound 51). Carbamate derivatives of the 2-amino function can be obtained by using

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2018203022 01 May 2018 chloroformates, such as benzylchlorofonnate. (See, for example, the preparation of Compound 58). The tertiary butyl carbamoyl function can also serve as a removable protecting group of the 2-amino function, (see for example the preparation of Compounds 219 and 224). When the 2-amino function of the compounds of the invention is already acylated or bears a carbamoyl group, then the 3-hydroxy group of the propanoic acid moiety can be subjected to acylation by reagents such as acetic anhydride. (See for example the preparation of Compound 217).

Alkylation of the 2-amino function is readily performed by condensing the compound bearing the 2-NH₂ group with an aldehyde to obtain a Schiff base intermediate which can be reduced, without isolation, to provide the TV-alkyl, arylalkyl or heteroaryl-alkyl compound. The procedure described for preparing Compound 234 can be generalized to make compounds of the invention where the 2-amino function bears an aryl( hydroxy)alkyl or heteroaryl(hydroxy)alkyl group.

Several compounds of the invention of Formula 2 can be obtained by derivatization of compounds of Formula 1, or by such modification of the synthetic routes leading to compounds of Formula 1 which will become readily apparent to those skilled in the art in light of the present disclosure. For example, compounds of Formula 2 where R₉ is OH or ORn and Rjo is alkyl can be made by using a “ketone” bearing the Ri₀ group, instead of the “aldehyde” in General Reaction Scheme 1.

Compounds of Formula 2 where the R₉ and Rio groups jointly form an oxime (NOH) group can be obtained by oxidizing the 3-hydroxyl group of the propanoic acid moiety to the ketone stage and reacting the resulting ketone with hydroxylamine.

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Another general synthetic route for making several compounds of Formula 2 is illustrated in Synthetic Scheme 3 adapted for synthesizing (R)-2amino-3-(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)propan-1 -one dihydrochloride, Compound 23 6. of the present invention. .

Compound 236

As it will be readily understood by those skilled in the art, for a more general synthetic route, such as the one shown in Reaction Shyntetic Scheme 3, the 4-pyridyl group can be substituted with an R* group (as defined in connection with Scheme 1) and the pyrrolidine can be substituted with amines of the formula NH(R**)₂ (R** defined as in connection with Reaction Scheme 1) to provide other compounds of Formula 2 analogous to compound 236 or to its enantiomer (5)-2-amino-3-(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)propan-1 -one dihydrochloride Compound 240.

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DETAILED DESCRIPTION OF THE SYNTHESIS OF PREFERRED COMPOUNDS (EXPERIMENTAL)

Preparation of compound 12

2-Isocyano-l-(pyrrolidin-l-yl)ethanone BLE 04098,

To stirred and cooled (0 °C) methyl isocyanoacetate (96 % technical grade, 5.0 g, 47.8 mmol) was slowly added in 0.75 h pyrrolidine (6.5 mL, 78 mmol). The mixture was stirred for 1.5 h with continued cooling and then concentrated. The resulting oil was co-evaporated twice from CH₂Cl₂:hexane to remove residual pyrrolidine. 2-Isocyano-l-(pyrrolidin-l-yl)ethanone BLE 04098 was obtained as a yellow solid (6.85 g, 98 % yield) and used in the next step without purification.

BLE 04098

MW: 138.17; Yield: 98 %; yellow solid; Mp (°C) = 73.9.

Al-NMR (CDC1₃, δ): 1.81-2.08 (m, 4H, 2xCH₂), 3.35-3.45 (m, 2H, -NCH₂), 3.50-3.60 (m, 2H, -NCH^, 4.23 (s, 2H, CH₂CO).

frfl/w-(4,5-DihydiO-5-(2,3-dihydiObenzo[Z?1 Γ1,41dioxin-6-yl)oxazol-4yl)(pyrrolidin-l-yl)methanone BLE 04100.

To a stirred and cooled (0°C) solution of potassium hydroxide (0.43 mg, 7.60 mmol) in MeOH (6.5 mL). were added successively l,4-benzodioxan-6carboxaldehyde (1.31 g, 7.96 mmol) and2-isocyano-l-(pyrrolidin-lyl)ethanone BLE 04098 (1.0 g, 6.57 mmol). The solution was stirred 3 h at 0°C and then concentrated. The residue was partitioned between EtOAc (100 mL)

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2018203022 01 May 2018 and water. The organic layer was combined with 2 additional EtOAc extracts (2 x 100 mL), washed with brine, dried over MgSO4, filtered and evaporated. Concentration afford to a crude product which was purified by column chromatography on silica (EtOAc) to yield, after evaporation and drying, to /razw-4,5-dihydro-5-(2,3-dihydiObenzo[0] [ 1,4]dioxin-6-yl)oxazol-4yl)(pyrrolidin-l-yl)methanone BLE 04100 as a colourless oil (1.76 g, 89 % yield).

(+/-)

BLE 04100

MW: 440.49; Yield: 89 %; colorless oil.

Ή-NMR (CDCIs, δ): 1.75-2.10 (m, 4H, 2xCH₂), 3.40-3.59 (m, 3H, 1.5xCH₂N),

3.85-4.00 (m, 1H, 0.5xCH₂N), 4.26 (s, 4H, CH₂O), 4.59 (dd, 1H, J= 7.5 Hz, J=

2.2 Hz, CH-N), 6.00 (d, 1H, J= 7.5 Hz, CH-O), 6.75-6.90 (m, 3H, ArH), 7.00 (d, 1H, J =22 Hz, CH=N).

/z777?5-(4,5-Dihydro-5-(4-methoxYphenyl)oxazol-4-yl)(pynOlidin-lyDmethanone SLA 07074.

To a stirred and cooled (0°C) solution of potassium hydroxide (0.37 g,

6.57 mmol) in methanol (30 mL) was added a mixture of 4-methoxybenzaldehyde (0.88 mL, 7.23 mmol) and 2-isocyano-l-(pyiTolidin-lyl)ethanone BLE 04098 (1.0 g, 6.57 mmol). The solution was stirred 4 h with continued cooling and then concentrated. The residue was partitioned between ethyl acetate and water. The organic layer was combined with additional ethyl acetate extracts, washed with aqueous sodium chloride and dried over MgSO4. Concentration afforded a crude product as a glassy solid. Flash chromatography

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2018203022 01 May 2018 over silica (ethyl acetate) yielded to b7Wi,s-(4,5-dihydro-5-(4methoxyphenyl)oxazol-4-yl)(pyrrolidin-l-yl)methanone SLA 07074 as a pale yellow solid (1.2 g, 90.5 %).

(+/-)

SLA 07074

MW: 274.32; Yield: 90.5 %; pale yellow solid; Mp (°C): 91.2. R/0.30 (EtOAc). . .

Ή-NMR (CDC1₃, δ): 1.75-2.08 (m, 4H, 2xCH₂), 3.40-3.58 (m, 3H, CH₂N), 3.52 (s, 3H, CH₃O), 3.88-3.98 (m, 1H, CH₂N), 4.59 (dd, 1H, J= 7.6 Hz, J= 22 Hz, CH-N), 6.06 (d, 1H, J= 7.6 Hz, CH-O), 6.90 (d, 2H, J= 8.7 Hz, ArH), 7.01 (d, 1H, J =22 Hz, CH=N), 7.25 (d, 2H, J= 8.7 Hz, ArH).

MS-ESI m/z (% rel. Int.): 275.1 ([MH]⁺, 10), 247.1 (100).

HPLC: Method A, detection UV 280 nm, SLA 07074 RT = 52 min, peak area 92 %.

DL-/Areo-2-Amino-3-hvdiOxv-3-(4-methoxyphenyl)-l-(pyrrolidin-l-yl)propan-

1-one hydrochloride SLA 07078.

To a stirred solution of trans-(fl,5-dihydro-5-(4-metlioxyphenyl)oxazol-4yl)(pyrrolidin-l-yl)methanone SLA 07074 (1.61 g, 5.93 mmol) in methanol (13 mL) was added hydrochloric acid (ImL). After heating at 50 °C for 3h the mixture reaction was concentrated and the resulting yellow oil was coevaporated twice with ethyl acetate before solidifying. Trituration (ethyl acetate) and drying afforded DL-//zreo-2-amino-3-hydroxy-3-(4WO 2006/081273

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oh o

(+/-)

SLA 07078

MW: 300.78; Yield: 93 %; white Solid; Mp (°C): 177.0.

‘H-NMR (CD₃OD, δ): 1.32-1.50 (m, 1H, 0.5xCH₂), 1.50-1.88 (m, 3H, 1.5xCH₂), 2.15-2.28 (m, 1H, CH₂N), 3.15-3.42 (m, 3H, 1.5xCH₂N), 3.79 (s, 3H, CH₃O), 4.06 (d, 1H, J= 9.2 Hz, CH-N), 4.78 (d, 1H, J= 9.2 Hz, CHO), 6.94 (d, 2H, J= 8.5 Hz, ArH), 7.34 (d, 2H, J= 8.5 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 24.8, 26.6, 47.2,47.6, 55.9, 59.6, 73.9, 115.0 (2xC), 128.9 (2xC), 132.5, 161.7, 166.4.

DL-//ireo-2-amino-3-(2,3-dihydiObenzor&irL41dioxin-6-yl)-3-hydroxy-l(pyrrolidin-l-yl)propan-l-one hydrochloride Compound 12.

To a stirred solution of ^a/w-4,5-dihydro-5-(2,3dihydrobenzo[b][l,4]dioxin-6-yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone BLE 04100 (1.74 g, 5.77 mmol) in methanol (15 mL) was added hydrochloric acid (1 mL). After heating at 50 °C for 3 h the mixture reaction was concentrated and the resulting yellow oil was co-evaporated twice with ethyl acetate before solidifying. Trituration (ethyl acetate) and drying afforded DL-//zreo-2-amino-3(2,3 -dihydrobenzo [A] [ 1,4] dioxin-6-yl)-3-hydroxy-1 -(pyrrolidin-1 -yl)propan-1 one hydrochloride Compound 12 as a white solid (1.85 g, 95 %).

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Compound 12

MW: 328.79; Yield: 95.0 %; White Solid; Mp (°C): 176.2.

^-NMR (CD₃OD, δ): 1.42-1.58 (m, 1H, 0.5xCH₂), 1.58-1.70 (m, 1H, 0.5xCH₂), 1.70-1.88 (m, 2H, CH₂), 3.20-3.45 (m, 4H, 2xN-CH₂), 4.06 (d, 1H, J = 9.1 Hz, CH-N), 4.25 (s, 2H, OCH₂), 4.75 (d, 1H, J= 9.2 Hz, CH-O), 4.89 (s, 2H, OCH₂), 6.82-6.95 (m, 3H, ArH).

¹³C-NMR (CD₃OD, δ): 24.9, 26.7, 47.3, 47.6, 59.5, 65.7, 73.6, 116.4, 118.3,

120.3,133.7,145.1, 145.6,166.4.

Preparation of Compound 18.

Method B:

To a stirred and cooled (0 °C) solution of potassium hydroxide (380 mg,

5.80 mmol) in MeOH (5 mL) were added successively aldehyde (5.80 mmol) and 2-isocyano-l-(pyrrolidin-l-yl)ethanone BLE 04098 (0.8 g, 5.8 mmol). The solution was stirred 3 h at 0 °C and then concentrated. The residue was partitioned between CH₂C1₂ (100 mL) and water. The organic layer was washed with brine, dried over MgSC>4, filtered and evaporated. Concentration afford to a crude product which was purified by column chromatography on silica (cyclohexane:EtOAc = 70:30 to 0:100) to yield, after evaporation and drying, to an intermediate oxazoline. To a stirred solution of oxazoline in methanol (15 mL) was added hydrochloric acid (1 mL, 12 mmol). After heating at 60 °C for 2 h, the mixture reaction was then concentrated and the resulting yellow oil was

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DL-//zreo-2-Amino-3 -(biphenyl-4-yl)-3-hydiOxy-1 -(p yrrolidin-1 - vDpropan-1 one hydrochloride Compound 18,

The compound was prepared according to method B with 4phenylbenzaldehyde (1.05 g, 5.78 mmol). DL-/7zreo-2-Amino-3-(biphenyl-4yl)-3-hydroxy-l-(pyiTolidin-l-yl)propan-l-one hydrochloride Compound 18 was obtained as a pale brown solid (0.55 g, 28 % yield).

OH 0

Compound 18

MW: 346.85; Yield: 28 %; Pale Brown Solid; Mp (°C): 197.3. ^-NMR (CD3OD, δ): 1.25-1.42 (m, 1H, 0.5xCH₂), 1.50-1.60 (m, 1H, 0.5xCH₂), 1.60-1.80 (m, 1H, 0.5xCH₂), 2.20-2.30 (m, 2H, N-CH₂), 3.15-3.30 (m, 2H, N-CH₂), 3.30-3.45 (1H, m, N-CH₂), 4.13 (d, 1H, J= 9.2 Hz, CH-N),

4.85-4.95 (m, 1H, CH-O), 7.32 -7.38 (m, 1H, ArH), 7.46 (dd, 2H, J = 7.1 Hz, J = 7.8Hz, ArH), 7.52 (d, 2H, J= 8.3 Hz, ArH), 7.58-7.70 (m, 4H, ArH). ¹³C-NMR (CD3OD, δ): 24.8, 26.5, 47.2,47.6, 59.5, 78.7, 127.9, 128.1, 128.2,

128.8, 130.0, 139.7, 141.6, 143.3, 166.3.

MS-ESI m/z (% rel. Int.): 311.2 ([MH]⁺, 60).

HPLC: Method A, detection UV 254 nm, Compound 18 RT = 4.50 min, peak area 99.9 %.

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Preparation of 2-isocvano derivatives: SLA 07116B, SLA 07116C, SLA 07118, SLA 07130A, SLA 07178, and SLA 07184A,

2-Isocyano-l-(piperidin-l-yl)ethanone SLA 07116B.

Prepared in accordance with Method B with methyl isocyanoacetate (2.46 g, 24.63 mmol) and piperidine (3.22 mL, 37.85 mmol). The reaction mixture was stirred 1 h at RT and then concentrated. The residue was dissolved in dichloromethane (50 mL) and the organic layer was washed with 10 % aqueous citric acid (2 x 25 mL), dried over MgSC>4, filtered and evaporated. 2-Isocyano- l-(piperidin-l-yl)ethanone SLA 07116B was obtained as an orange solid (3.13 g, 83 % yield).

SLA 07116B

MW: 152.19; Yield: 83 %; Orange Solid; Mp (°C): 81.6.

^NMR (CDC1₃, δ): 1.56-1.74 (m, 6H, CH₂C), 3.33 (t, 2H, 5.7 Hz, CH₂N),

3.58 (t, 2H, J= 5.7 Hz, CH₂N), 4.29 (s, 2H, CH₂CO).

/e/7-Butyl 4-(2-isocyanoacetyl)piperazine-l-carboxylate SLA 07116C.

Prepared in accordance with Method B with methyl isocyanoacetate (2.51 g, 25.29 mmol) and piperazine-1-carboxylic acid tert-butyl ester (6.28 g, 33.85 mmol. The reaction mixture was stirred 1 h at RT and then concentrated. The residue was dissolved in dichloromethane (50 mL) and the organic layer was washed with 10 % aqueous citric acid (2 x 25 mL), dried over MgSO4, filtered

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07116C was obtained as a colorless oil (0.41 g, 6.5 % yield).

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NBoc

SLA07116C

MW: 253.14; Yield: 6.5 %; Colorless oil.

^-NMR (CDC1₃, δ): 1.47 (s, 9H, tBu), 3.38 (t, 2H, J= 5.3 Hz, CH₂N), 3.45

3.53 (m, 4H, CH₂N), 3.62 (t, 2H, J= 5.5 Hz, CH₂N), 4.32 (s, 2H, CH₂CO).

2-Isocyano-1 -morpholinoethanone SLA 07118.

Prepared in accordance with Method B with methyl isocyanoacetate (2.51 g, 25.30 mmol) and morpholine (3.30 mL, 38.05 mmol). The reaction mixture was stirred 24 h at RT and then concentrated. The residue was dissolved in dichloromethane (50 mL) and the organic layer was washed with 10 % aqueous, citric acid (2 x 25 mL), dried over MgSO.4, filtered and evaporated. 2-Isocyano-

1-morpholinoethanone SLA 07118 was obtained as a brown oil (2.28 g, 58 % yield).

SLA 07118

MW: 154.17; Yield: 58 %; Brown Oil.

Ry·: 0.20 (EtOAc:cyclohexane = 50:50).

^-NMR (CDCI3, δ): 3.42 (t, 2H, J= 4.9 Hz, CH₂N), 3.65 (t, 2H, /=5.1 Hz,

CH₂N), 3.73 (t, 4H, J= 5.0 Hz, CH₂O), 4.31 (s, 2H, CH₂CO).

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2-Isocyano-l-thiomorpholinoethanone SLA 07130A,

Prepared in accordance with Method B with methyl isocyanoacetate (2.50 g, 25.28 mmol) and thiomorpholine (4.25 mL, 37.85 mmol). The reaction mixture was stirred 22 h at RT and then concentrated. The residue was dissolved in dichloromethane (50 mL) and the organic layer was washed with 10 % aqueous citric acid (2 x 25 mL), dried over MgSO₄, filtered and evaporated. 2-Isocyano-l-thiomorpholinoethanone SLA 07130A was obtained as a yellow solid (3.05 g, 71 % yield).

SLA07130A

MW: 170.23; Yield: 71 %; Yellow Solid; Mp (°C): 144.4.

Ry: 0.35 (EtOAc:cyclohexane = 50:50).

Ή-NMR (CDC1₃, δ): 2.68 (m, 4H, 2xCH₂S), 3.67 (m, 2H, N-CH₂), 3.90 (m, 2H, N-CH₂), 4.31 (s, 2H, COCH₂).

2-Isocyano-l-(27T-pyrrol-l(517)-yl)ethanohe SLA 07178.

Prepared in accordance with Method B with methyl isocyanoacetate (1.00 g, 10.10 mmol) and dihydro-lTApyrrole (1.01 mL, 15.15 mmol). The reaction mixture was stirred 5 h at 50 °C and concentrated. The residue was dissolved in dichloromethane (50 mL) and the organic layer was washed with 10 % aqueous citric acid (2 x 25 mL), dried over MgSO₄, filtered and evaporated. 2-Isocyano- l-(2H-pyrrol-l(577)-yl)ethanone SLA 07178 was obtained (1.0 g, 73 % yield) as a yellow solid.

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SLA 07178

MW: 136.15; Yield: 73 %; Yellow Solid.

Ry: 0.3 5 (EtOAc: cyclohexane = 50:50).

Y-NMR (CDC1₃, δ): 4.23 (s, 4H, 2xCI-I₂N), 4.31 (s, 2H, CH₂N), 5.80-5.86 (m, 1H, CH=C), 5.90-5.95 (m, 1H, CI-I=C).

M/V-Diethyl-2-isocyanoacetamide SLA 07184A,

Prepared in accordance with Method B with methyl isocyanoacetate (2.50 g, 25.29 mmol) and diethylamine (1.96 mL, 37.94 mmol). The reaction mixture was stirred 5 h at 50 °C and concentrated. The residue was dissolved in dichloromethane (50 mL) and the organic layer was washed with 10 % aqueous citric acid (2 x 25 mL), dried over MgSO₄, filtered and evaporated. N,NDiethyl-2-isocyanoacetamide SLA 07184A was obtained (1.213 g, 34 % yield) as a brown oil.

SLA07184A

MW: 140.18; Yield: 34 %; Brown Oil.

Ry: 0.35 (EtOAc:cyclohexane = 50:50).

^!H-NMR (CDC1₃, δ): 1.15-1.26 (m, 6H, CH₃), 3.21-3.30 (m, 2H, CH₂N), 3.38. 3.45 (m, 2H, CH₂N), 4.26 (s, 2H, CH₂CO).

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Preparation of oxazolines: BLE 04110B, SLA 07122A, SLA 07124A, SLA 07124B, SLA 07132, BLE 04110A, Compound 19, BLE 04124A, BLE 04124B, BLE 04124C, BLE 04124D, BLE 04130B, BLE 04130C, BLE 04130D, BLE 04136B, BLE 04136C, BAL 01016, BLE 04136D, BAL 01014, SLA 07194A, SLA 07174, BAL 01028A, BLA 01028B, SLA 07158 and SLA 07180.

/rans-(4,5-Dihydro-5-(pyridin-3-vl)oxazol-4-yl)(pvrrolidin-l-yl)methanone BLE04110B.

General Method D for oxazolines formation:

To a stirred and cooled (0 °C) solution of potassium hydroxide (0.55 g,

9.80 mmol) in methanol (10 mL) were added a mixture of 3-pyridine carboxaldehyde (1.03 mL, 10.84 mmol) and 2-isocyano-l-(pyrrolidin-lyl)ethanone BLE 04098 (1.50 g, 10.86 mmol). The solution was stirred 3 h at 0 °C and then concentrated. The residue was partitioned between ethyl acetate (100 mL) and water. The organic layer was combined with two additional ethyl acetate extracts (2x100 mL), washed with aqueous sodium chloride and dried over MgSCL, filtered and evaporated. Concentration afforded a crude product which was purified by column chromatography on silica (CH₂Cl₂:MeOH = 98:2) to yield to trans-(4,5-dihydro-5-(pyridin-3-yl)oxazol-4-yl)(pyrrolidin-lyl)methanone BLE 04110B (0.95 g, 39 % yield) as a pale yellow pale solid.

(+/-)

BLE04110B

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MW: 245.28; Yield: 39 %; Yellow Pale Solid; Mp (°C): 107.0.

’H-NMR (CDCI3, δ): 1.78-2.10 (m, 4H, 2xCH₂), 3.40-3.61 (in, 3H, CH₂N),

3.90-4.04 (ni, 1H, CH₂N), 4.59 (dd, 1H, 7.7 Hz, J= 2.2 Hz, CH-N), 6.21 (d,

1H, J= 7.7 Hz, CH-O), 7.04 (d, 1H, J= 2.2 Hz, O-CH=N), 7.33 (m, 1H, ArH), 7.64 (m, 1H, ArH), 8.59 (d, 2H, J= 2.8 Hz, ArH).

¹³C-NMR (CDCI3, δ): 24.2, 26.0, 46.4,46.6, 75.7, 79.3, 123.7, 133.5,135.3, 147.6,149.9, 155.2, 166.2. '

Zm//5-(4,5-Dihydro-5-(pyridin-4-yl)oxazol-4-yl)(piperidin-1 -yl)methanone SLA 07122A.

SLA 07122A was prepared in accordance with method method D using 2isocyano-l-(piperidin-l-yl)ethanone (0.4 g, 26.3 mmol), potassium hydroxide (0.15 g, 26.7 mmol) in methanol (5 mL) and pyridine-4-carbaldehyde (0.37 mL,

40.9 mmol). The solution was stirred 20 h at 0 °C. ira/w-(4,5-Dihydro-5(pyridin-4-yl)oxazol-4-yl)(piperidin-l-yl)methanone SLA 07122A was obtained as a yellow solid (0.353 g, 52 % yield).

SLA07122A

MW: 259.30; Yield: 52 %; Yellow Solid; Mp (°C): 111.7.

R_z: 0.80 (MeOH:CH₂Cl₂ = 10:90).

^-NMR (CDCI3, δ): 1.55-1.78 (m, 6H, 3xCH₂), 3.45-3.60 (m, 2H, CH₂N),

3.70-3.85 (m, 2H, CH₂N), 4.60 (dd, 1H, J= 7.8 Hz, J= 2.3 Hz, CH-N), 6.27 (d,

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1H, 7= 7.8 Hz, CH-O), 7.01 (d, 1H, J= 2.3 Hz, CH=N), 7.23 (dd, 2H, J= 4.5

Hz, J= 1.6 Hz, ArH), 8.61 (dd, 2H, J= 4.5 Hz, 7= 1.5 Hz, ArH).

/rmzs-(4,5-Dihvdro-5-(pvridin-4-yl)oxazol-4-yl)(morpholino)methanone SLA 07124A.

SLA 07118 was prepared in accordance with method D using 2-isocyano-

1-morpholinoethanone (0.40 g, 25.95 mmol), potassium hydroxide (0.146 g, 26.0 mmol) in methanol (5 mL) and pyridine-4-carbaldehyde (0.36 mL, 40.4 mmol). The solution was stirred 22 h at 0 °C. trans-(A,5-Dihydro-5-(pyridin-4yl)oxazol-4-yl)(morpholino)methanone SLA 07124A was obtained as a yellow solid (0.168 g, 25 % yield).

O-A ⁰ </° (+/-)

SLA07124A

MW: 261.28; Yield: 25 %; Yellow Solid; Mp (°C): 90.5.

Ry·: 0.30 (EtOAc:cyclohexane = 20:80).

^-NMR (CDC1₃, δ): 3.46-4.02 (m, 8H, 2xCH₂O, 2xCH₂N), 4.56 (dd, 1H, J=

7.8 Hz, J= 2.3 Hz, CH-N), 6.27 (d, 1H, J= 7.9 Hz, CH-O), 7.02 (d, 1H, J= 2.3 Hz, CH=N), 7.24 (dd, 2H, J= 4.6 Hz, J= 1.4 Hz, ArH), 8.63 (dd, 2H, J= 4.5 Hz, 7= 1.6 Hz, ArH).

/m/^-(4,5-Dihydro-5-(pyridin-4-yl)oxazol-4-yl)(4-te77-butyloxycarbonylpiperazin-l-yl)methanone SLA 07124B.

SLA 07124B was prepared in accordance with method D using iezY-butyl

4-(2-isocyanoacetyl)piperazine-l-carboxylate SLA 07116C (0.41 g, 16.20

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:oc (+/-)

SLA07124B

MW: 360.41; Yield: 58 %; Pale Yellow Solid; Mp (°C): 157.2°C.

Ή-NMR (CDC1₃, δ): 1.47 (s, 9H, tBu), 3.25-4.02 (m, 8H, CH₂N), 4.58 (dd, 1H, J- 7.8 Hz, J= 2.3 Hz, CH-N), 6.27 (d, 1H, J= 7.8 Hz, CH-O), 7.01 (d, 1H, J-

2.3 Hz, CH=N), 7.24 (dd, 2H, J= 4.6 Hz, J= 1.4 Hz, ArH), 8.62 (dd, 2H, J=

4.5 Hz, J = 1.6 Hz, ArH).

frans-i4A-Dihydro-5-(pyridin-4-yl)oxazol-4-yl)(thiomorpholino)methanone SLA 07132. .

SLA 07132 was prepared in accordance with method D using 2-Isocyano-

1-thiomorpholinoethanone SLA 07130A (0.752 g, 4.41 mmol), potassium hydroxide (0.250 g, 4.45 mmol) in methanol (10 mL) and pyridine-4carbaldehyde (0.436 mL, 4.85 mmol). The solution was stirred 24 h at 0 °C. ira/w-(4,5-Dihydro-5-(pyridin-4-yl)oxazol-4-yl)(thiomorpholino)methanone SLA 07132 was obtained as a yellow foam (1.01 g, 83 %).

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SLA 07132

MW: 277.35; Yield: 83 %; Yellow Foam.

R_z: 0.80 (MeOH:CH₂Cl₂ = 10:90).

^-NMR (CDC1₃, δ): 2.53-2.92 (m, 4H, 2xCH₂), 3.58-3.70 (m, 1H, CH₂N),

3.78-3.88 (m, 1H, CH₂N), 4.15-4.30 (m, 2H, CH₂N), 4.56 (dd, J= 7.8 Hz, J=

2.3 Hz, 2H, CH-N), 6.27 (d, 1H, J= 7.8 Hz, CH-O), 7.02 (d, 1H, J= 2.3 Hz, N=CH-O), 7.22 (d, 2H, J= 6.1 Hz, ArH), 8.61 (dd, 2H, J= 6.1 Hz, ArH). ¹³C-NMR (CDCI3, δ): 27.3, 28.0,45.4,48.6, 74.9, 79.6,120.0 (2xC), 148.5,

150.3 (2xC), 154.8, 166.2.

fraws-(4A-Dihydro-5-(pyridin-2-yl)oxazol-4-yl)(pvrrolidin-l-yl)methanone BLE04110A.

BLE 04110A was prepared in accordance with method D using 2pyridine carboxaldehyde (1.02 mL, 10.84 mmol). Tzww-(4,5-dihydro-5(pyridin-2-yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone BLE 04110A was obtained as a yellow pale oil (0.45 g, 19 % yield).

(+/-)

BLE04110A

MW: 245.28; Yield: 19 %; Yellow Pale Oil.

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2018203022 01 May 2018 ^!H-NMR (CDC1₃, δ): 1.73-2.08 (m, 4H, 2xCH₂), 3.35-3.70 (m, 3H, CH₂N),

3.85-4.00 (in, 1H, CH2N), 5.05 (dd, 1H, J= 6.9 Hz, J= 2.2 Hz, CH-N), 6.18 (d, 1H, J= 6.9 Hz, CH-O), 7.02 (d, 1H, J= 2.1 Hz, O-CH=N), 7.25 (m, 1H, ArH),

7.43 (d, 1H, J= 7.8 Hz, ArH), 7.69 (dt, 1H, J= 7.8 Hz, /= 1.8 Hz ArH), 8.62 (m, 1H, ArH).

¹³C-NMR (CDCI3, δ): 24.2, 25.9,46.3,46.5, 73.4, 81.3, 121.5, 123.2, 136.8,

149.8,154.8, 158.0, 166.9.

/raKs-(4,5-Dihydro-5-(pyridin-4-yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone Compound 19.

SLA 07092 was prepared in accordance with method D using pyridine-4carbaldehyde (1.88 mL, 19.76 mmol), KOH (1.01 g, 18.00 mmol) in methanol (18 mL) and 2-isocyano-l-(pyrrolidin-l-yl)ethanone BLE 04098 (2.73 g, 19.76 mmol). The residue was partitioned between ethyl acetate (200 mL) and water (150 mL). The organic layer was combined with additional ethyl acetate extracts (2 x 150 mL), washed with aqueous sodium chloride (2 x 150 mL) and dried over MgSO4, filtered and evaporated. Trans-(A,5-dihydro-5-(pyridin-4yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone Compound 19 was obtained as a white solid (4.32 g, 98 % yield).

(+/-)

Compound 19

MW: 245.28; Yield: 98 %; White Solid; Mp (°C) = 69.2.

R_z: 0.65 (MeOH:CH₂Cl₂ = 10:90).

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Ή-NMR (CDC1₃, δ): 1.78-2.06 (m, 4H, 2xCH₂), 3.44-3.60 (m, 3H, CH₂N),

3.90-4.01 (m, 1H, CH₂N), 4.52 (dd, 1H, J= 7.9 Hz, J= 2.2 Hz, CH-N), 6.19 (d, J= 7.9 Hz, 1H, CH-O), 7.03 (d, 1H, J = 2.2 Hz, N=CH-O), 7.24 (dd, 2H,

4.5 Hz, J= 1.5 Hz, ArH), 8.61 (dd, 2H, 4.5 Hz, J= 1.5 Hz, ArH).

/m«5^,-(4,5-Dihydro-5“(thiophen-3-yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone BLE 04124A.

BLE 04124A was prepared in accordance with method D using thiophen-

3-carboxaldehyde (0.475 mL, 5.42 mmol), KOH (0.276 mg, 4.92 mmol) in methanol (5 mL) and 2-isocyano-l-(pyrrolidin-l-yl)ethanone BLE 04098 (0.75 g, 5.43 mmol). After work-up the residue obtained was recristallized from ethyl acetate to obtain after filtration trans-(4,5-dihydro-5-(thiophen-3-yl)oxazol-4yl)(pyrrolidin-l-yl)methanone BLE 04110A as a yellow pale solid (0.498 g, 40 % yield).

BLE 04124A

MW: 250.32; Yield: 40.5 %; Yellow Pale Solid; Mp (°C): 105.9.

Ή-NMR (CDC1₃, δ): 1.78-2.10 (m, 4H, CH₂), 3.42-3.61 (m, 3H, CH₂N), 3.904.02 (m, 1H, CH₂N), 4.63 (dd, 1H, J= 7 A Hz, J= 2.2 Hz, CH-N), 6.20 (d, 1H, J= 7.4 Hz, CH-O), 6.98 (d, 1H, J= 2.2 Hz, O-CH=N), 7.03 (dd, 1H, J= 5.0 Hz, J= 1.3 Hz, CH=C), 7.30 (dt, 1H, J= 3.0 Hz, J= 1.3 Hz, CH=C), 7.36 (dd, 1H, J= 5.0 Hz, J= 3.0 Hz, CH=C).

¹³C-NMR (CDC1₃, δ): 24.2, 26.0,46.4,46.6, 74.6, 77.9, 122.7, 125.1, 127.3,

140.4, 155.3, 166.7.

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MS-ESI m/z (% rel. Int.): 251.0 ([MH]⁺, 17), 223 (40), 179.9 (60), 151.9 (63),

123.9 (100).

HPLC: Method A, detection UV 254 nm, BLE 04124A, RT = 4.4 min, peak area 98.0 %.

/rans^,-(4,5-Dihydro-5-(thiophen-2-yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone BLE 04124B.

BLE 04124B was prepared in accordance with method D using thiophen-

2-carboxaldehyde (0.507 mL, 5.42 mmol), KOH (0.276 mg, 4.92 mmol) in methanol (5 mL) and 2-isocyano-l-(pyrrolidin-l-yl)ethanone BLE 04098 (0.75 g, 5.43 mmol). After work-up the residue obtained was purified by column chromatography (EtOAc) to led after evaporation to trans-(4,5-dihydro-5-( thiophen-2-yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone BLE 04124B as a yellow pale solid (0.713 g, 58 % yield).

(+/-)

BLE 04124B

MW: 250.32; Yield: 58 %; Yellow Pale Solid; Mp (°C): 71.3.

Ή-NMR (CDC1₃, δ): 1.78-2.10 (m, 4H, CH₂), 3.42-3.62 (m, 3H, CH₂N), 3.904.03 (m, 1H, CH₂N), 4.76 (dd, 1H, 7.3 Hz, J= 2.2 Hz, CH-N), 6.37 (d, 1H,

7.3 Hz, CH-O), 6.96 (d, 1H, J= 2.2 Hz, O-CH=N), 7.00 (dd, 1H, J= 5.0 Hz, J = 3.5 Hz, CH=C), 7.11 (d, 1H, J= 3.1 Hz, CH=C), 7.33 (dd, 1H, J= 5.0 Hz, J = 0.7 Hz, CH=C).

¹³C-NMR (CDC1₃, δ): 24.2, 26.0, 46.4,46.6, 75.5, 77.6,126.3 (2xC), 127.1, 142.0, 154.9, 166.3.

MS-ESI m/z (% rel. Int.): 251.0 ([MH]⁺, 15), 223 (100).

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HPLC: Method A, detection UV 254 nm, BLE 04124B, RT = 3.8 min, peak area > 90 %.

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Zmn 5^,-(4,5-Dihydro-5-(thiazol-2-yl)oxazol-4-yl)(pyrrolidin-1 -yDmethanone BLE 04124C.

BLE 04124C was prepared in accordance with method D using 2thiazolecarboxaldehyde (0.476 mL, 5.42 mmol), KOH (0.276 mg, 4.92 mmol) in methanol (5 mL) and 2-isocyano-l-(pyrrolidin-l-yl)ethanone BLE 04098 (0.75 g, 5.43 mmol). After work-up the residue obtained was purified by column chromatography (EtOAc) to led after evaporation to trans-(4,5-dihydro-

5-(thiazol-2-yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone BLE 04124C as a colourless oil (0.564 g, 45.5 % yield).

(+/-)

BLE 04124C

MW: 251.3; Yield: 45.5 %; colourless Oil.

Ή-NMR (CDC1₃, δ): 1.80-2.10 (m, 4H, CH₂), 3.47-3.70 (m, 3H, CH₂N), 3.914.02 (m, 1H, CH₂N), 5.18 (dd, 1H, J= 6.4 Hz, J= 2.2 Hz, CH-N), 6.40 (d, 1H, J= 6.4 Hz, CH-O), 6.97 (d, 1H, J= 2.2 Hz, O-CH=N), 7.38 (d, 1H, J= 3.3 Hz, CH=C), 7.81 (d, 1H, 3.3 Hz, CH=C).

¹³C-NMR(CDC1₃, δ): 24.2, 26.0,46.4,46.5, 73.7, 78.2, 120.1, 143.3, 154.3,

166.1, 168.2.

MS-ESI m/z (% rel. Int.): 252.0 ([MH]⁺, 18), 225 (30), 198.9 (37), 153.9 (48), 143.0 (100).

HPLC: Method A, detection UV 254 nm, BLE 04124C, RT = 3.5 min, peak area > 90 %.

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2018203022 01 May 2018 /raws-(5-(Benzor61thiophen-3-vl)-4.,5-dihydrooxazol-4-yiypyiTolidm-lvDmethanone BLE 04124D.

BLE 04124D was prepared in accordance with method D using thianaphtene-3-carboxaldehyde (0.88 g, 5.42 mmol), KOH (0.276 mg, 4.92 mmol) in methanol (5 mL) and 2-isocyano-l-(pyrrolidin-l-yl)ethanone BLE 04098 (0.75 g, 5.43 mmol). After work-up the residue obtained was purified by column chromatography (EtOAc) to led after evaporation to ira/w-(5(benzo[0]thiophen-3-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-l-yl)methanone BLE 04124D as a white solid (1.12 g, 75.5 % yield).

BLE04124D

MW: 300.38; Yield: 75.5 %; White Solid; Mp (°C); 92.2.

^-NMR (CDC1₃, δ): 1.75-2.08 (m, 4H, CH₂), 3.36-3.49 (m, 1H, CH₂N), 3.503.62 (m, 1H, CH₂N), 3.89-4.00 (m, 1H, CH₂N), 4.75 (dd, 1H, J= 7.6 Hz, J=

Hz, CH-N), 6.54 (d, 1H, J= 7.6 Hz, CH-O), 7.08 (d, 1H, J - 2.2 Hz, OCH=N), 7.35 (m, 2H, ArH), 7.45 (s, 1H, C-CH-S), 7.67-7.75 (m, 1H, ArH), 7.84-7.92 (m, 1H, ArH).

¹³C-NMR (CDCI3, δ): 24.2, 26.0,46.5,46.6, 73.3, 77.7,121.8, 123.1, 124.1,

124.6, 124.8, 134.0, 136.4, 141.0, 155.4, 166.6.

MS-ESI m/z (% rel. Int.): 301.0 ([MH]⁺, 30), 273.0 (100).

HPLC: Method A, detection UV 254 nm, BLE 04124D, RT = 4.2 min, peak area 92.0 %.

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2018203022 01 May 2018 im/i.sⁱ-(5-(Furan-3-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidm-1 -vl)methanone BLE 04130B.

BLE 04130B was prepared in accordance with method D using 3furaldehyde (0.453 g, 5.42 mmol), KOH (0.276 mg, 4.92 mmol) in methanol (5 mL) and 2-isocyano-l-(pyrrolidin-l-yl)ethanone BLE 04098 (0.75 g, 5.43 mmol). After work-up the residue was washed with a minimum of ethyl acetate to led, after filtration and drying, to iran5-(5-(furan-3-yl)-4,5-dihydrooxazol-4yl)(pyrrolidin-l-yl)methanone BLE 04130B as a white solid (0.837 g, 72.5 % yield).

(+/-)

BLE 04130B

MW: 234.25; Yield: 72.5 %; White Solid; Mp (°C): 136.7.

^-NMR (CDC1₃, δ): 1.80-2.10 (m, 4H, CH₂), 3.47-3.58 (m, 3H, CH2N), 3.914.02 (m, 1H, CH2N), 4.61 (dd, 1H, J= 7.3 Hz, J= 2.1 Hz, CH-N), 6.10 (d, 1H, J= 7.3 Hz, CH-O), 6.36 (dd, 1H, J= 1.6 Hz, J= 0.6 Hz, CH=C), 6.95 (d, 1H, J = 2.1 Hz, O-CH=N), 7.44 (t, 1H, J= 1.6 Hz, OCH=C); 7.50 (d, 1H, J= 0.6 Hz, OCH=C).

¹³C-NMR (CDCI3, δ): 24.2, 26.0, 46.4,46.6, 70.0, 74.0,108.1, 124.0, 140.4,

144.2, 155.3, 166.6.

/m»s^,-(4,5-Dihydro-5-(naphthalen-3-yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone BLE 04130C.

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BLE 04130C was prepared in accordance with method D using 2naphtaldehyde (0.847 g, 5.42 mmol), KOH (0.276 mg, 4.92 mmol) in methanol (5 mL) and 2-isocyano-l-(pyrrolidin-l-yl)ethanone BLE 04098 (0.75 g, 5.43 mmol). After work-up the residue was washed with a minimum of ethyl acetate to led, after filtration and drying, to trans-(4,5-dihydro-5-(naphthalen-3yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone BLE 04130C as a white solid (0.791 g, 54.5 % yield).

MW: 294.35; Yield: 54.5 %; White Solid; Mp (°C): 117.9.

^!H-NMR (CDC1₃, δ): 1.78-2.07 (m, 4H, CH₂), 3.37-3.49 (m, 1H, CH₂N), 3.493.61 (m, 2H, CH₂N), 3.88-3.99 (m, 1H, CH₂N), 4.67 (dd, 1H, J= 7.7 Hz, J=

2.2 Hz, CH-N), 6.31 (d, 1H, J= 7.7 Hz, CH-O), 7.10 (d, 1H, J= 2.2 Hz, OCH=N), 7.38 (dd, 1H, J= 8.5 Hz, J= 1.7 Hz, ArH); 7.45-7.54 (m, 2H, ArH),

7.79-7.90 (m, 4H, ArH).

¹³C-NMR(CDC1₃, δ): 23.8, 25.7,46.1,46.2,75.3, 81.4,122.7, 124.9, 126.1,

126.2, 127.4, 127.7, 128.7, 132.8, 132.9, 136.5, 155.2,166.4.

MS-ESI m/z (% rel. Int.): 295.1 ([MH]⁺, 40), 267.1 (100).

HPLC: Method A, detection UV 254 nm, BLE 04130C, RT = 4.2 min, peak area 92.0 %.

/ran5^,-(4,5-Dihydro-5-(naphthalen-4-yl)oxazol-4-yl)(pyrrolidin-l-yI)methanone BLE 04130D.

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BLE 04130D was prepared in accordance with method D using 1naphtaldehyde (0.736 mL, 5.42 mmol), KOH (0.276 mg, 4.92 mmol) in methanol (5 mL) and 2-isocyano-l-(pyrrolidin-l-yl)ethanone BLE 04098 (0.75 g, 5.43 mmol). After work-up the residue was purified by column chromatography on silica (EtOAc:cyclohexane = 80:20 to 90:10) to led, after evaporation, to \rans-(f,5-dihydro-5-(naphthalen-4-yl)oxazol-4-yl)(pyrrolidin- l-yl)methanone BLE 04130D as a colorless gum (0.850 g, 58.5 % yield).

BLE 04130D

MW: 294.35; Yield: 58.5 %; Colorless gum.

Ή-NMR (CDC1₃, δ): 1.75-2.02 (m, 4H, CH₂), 3.25-3.37 (m, 1H, CH₂N), 3.52-

3.67 (m, 2H, CH₂N), 3.82 -3.93 (m, 1H, CH₂N), 4.62 (dd, 1H, J= 7.0 Hz, J= 2.0 Hz, CH-N), 6.89 (d, 1H, J= 7.0 Hz, CH-O), 7.16 (d, 1H, J= 2.0 Hz, OCH=N), 7.44-7.58 (m, 4H, ArH), 7.80-7.90 (m, 3H, ArH).

¹³C-NMR(CDC1₃, δ): 24.2, 25.9,46.5 (2xC), 75.3, 79.2, 122.5, 123.0,125.4, 126.0, 126.8, 128.7, 129.0, 129.9, 133.9, 135.5, 155.5,166.9.

MS-ESI m/z (% rel. Int.): 295.1 ([MH]⁺, 50), 267.1 (100).

HPLC: Method A, detection UV 254 nm, BLE 04BOD, RT = 4.2 min, peak area 95.0 %.

irans-(4,5-Dihydro-5-(quinolin-2-yl)oxazol-4-yl)(pvrrolidin-l-yr)methanone BLE 04136B.

BLE 04136B was prepared in accordance with method D using 2quinoline carbaldehyde (0.852 g, 5.42 mmol), KOH (0.276 mg, 4.92 mmol) in

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2018203022 01 May 2018 methanol (5 mL) and 2-isocyano-l-(pyrrolidin-l-yl)ethanone BLE 04098 (0.75 g, 5.43 mmol). After work-up the residue was purified by column chromatography on silica (EtOAc) to led, after evaporation, to irazi5-(4,5dihydro-5-(quinolin-2-yl)oxazol-4-yl)(pyrrolidin-1 -yl)methanone BLE 0413 6B as a yellow pale solid (0.966 g, 60.3 % yield).

(+/-)

BLE 04136B

MW: 295.34; Yield: 60.3 %; Yellow Pale Solid; Mp (°C): 93.8.

’H-NMR (CDC1₃, δ): 1.85-2.10 (m, 4H, CH₂), 3.50-3.66 (m, 2H, CH₂N), 3.67-

3.80 (m, 1H, CH₂N), 3.92-4.03 (m, 1H, CH₂N), 5.32 (dd, 1H, J= 7.8 Hz, J=

2.1 Hz, CH-N), 6.31 (d, 1H, J= 7.8 Hz, CH-O), 7.06 (d, 1H, J= 2.1 Hz, OCH=N), 7.51-7.60 (m, 2H, ArH); 7.72 (t, 1H, J= 8.4 Hz, ArH), 7.83 (t, 1H, J=

8.1 Hz, ArH), 8.07 (d, 1H, J= 8.4 Hz, ArH), 8.20 (d, 1H, J= 8.4 Hz, ArH). ¹³C-NMR (CDC1₃, δ): 24.3, 26.2,46.5,46.7, 73.0, 82.1,119.2, 126.9, 127.7,

127.8, 129.5, 129.9, 137.2, 147.7, 155.0, 158.2, 167.3.

/m/?3'-(4,5-Dihydro-5-(isoquinolin-4-yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone BLE 04136C.

BLE 04136C was prepared in accordance with method D using 4quinoline carbaldehyde (0.852 g, 5.42 mmol), KOH (0.276 mg, 4.92 mmol) in methanol (5 mL) and 2-isocyano-l-(pyrrolidin-l-yl)ethanone BLE 04098 (0.75 g, 5.43 mmol). After work-up the residue was washed with a minimum of EtOAc to led, after filtration and drying, to irans-(4,5-dihydro-5-(isoquinolin-4WO 2006/081273

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(+/-)

BLE 04136C

MW: 295.34; Yield: %; White Solid; Mp (°C): 152.0.

Ή-NMR (CDC1₃, δ): 1.85-2.08 (m, 4H, CH₂), 3.28-3.40 (m, 1H, CH₂N), 3.54-

3.69 (m, 2H, CH₂N), 3.84-3.95 (m, 1H, CH₂N), 4.57 (dd, 1H, J= 6.8 Hz, J=

2.1 Hz, CH-N), 6.93 (d, 1H, J= 6.8 Hz, CH-O), 7.15 (d, 1H, J= 2.1 Hz, 0CH=N), 7.41 (d, 1H, J= 4.5 Hz, ArH); 7.59 (m, 1H, ArH), 7.76 (m, 1H, ArH), 7.91 (d, 1H, 8.3 Hz, ArH), 8.16 (d, 1H, J= 8.3 Hz, ArH), 8.92 (d, 1H, J=

4.5 Hz, ArH).

¹³C-NMR (CDC1₃, δ): 24.2, 25.9,46.7 (2xC), 75.6, 77.8, 116.4, 123.1, 124.8,

127.5, 129.6, 130.5, 145.5, 148.4, 150.3, 155.0, 166.0.

/^725-(4,5-Dihydro-5-(quinolin-3-yl)oxazol-4-yl)(pvrrolidin-l-yl)methanone BAL 01016.

To a stirred and cooled (0 °C) solution of KOH (0.31 g, 5.43 mmol) in 5 mL MeOH were added successively quinoline-3-carboxaldehyde (0.85 g, 5.43 mmol) and 2-isocyano-l-pyrrolidin-l-yl-ethanone BLE 04134 (0.75 g, 5.43 mmol). The mixture was stirred at 0 °C until precipitation and concentrated. The mixture was partitioned between EtOAc (50 mL) and H₂O (25 mL). The aqueous layer was extracted twice with EtOAc (25 ml). The EtOAc fractions were combined, washed twice with brine (2x25 mL), dried over MgSO₄ and filtered. After evaporation and drying iraws-(4,5-dihydro-5-(quinolin-3WO 2006/081273

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BAL 01016

MW: 295.34; Yield: 60 %; White Solid; Mp (°C): 144.4.

R/’: 0.15 (EtOAc).

^!H NMR (CDCI3, δ): 1.75-2.10 (m, 4H, 2xCH₂), 3.40-3.62 (m, 3 H, CH₂N),

3.90-4.05 (m, 1 H, CH₂N), 4.70 (dd, 1H, J= 7.8 Hz, J= 2.2 Hz, CH-N), 6.40 (d, 1H, 7= 7.8 Hz, CH-O), 7.10 (d, 1H, 7=2.2 Hz, OCH=N), 7.58 (dt, 1H,7= 1.1 Hz, J= 8.0 Hz, ArH)), 7.73 (dt, 1H, 7= 1.4 Hz, 7= 6.9 Hz, ArH), 7.83 (dd, 1H, 7= 1.2 Hz, 7= 8.2 Hz, ArH), 8.12 (m, 2H, ArH), 8.87 (d, 1H, 7= 2.2 Hz, ArH). ¹³C-NMR (CDCI3, δ): 24.2, 26.0,46.6,46.6, 75.8, 79.7, 127.3, 127.5, 127.9, 129.4,130.0, 132.3, 133.2, 148.1, 148.4, 155.3, 166.2.

MS-ESI m/z (% rel. Int.): 296.1 ([MH]⁺, 5), 314.1 (100).

fmz/3^,-(5-(Furan-2-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1 -yDmethanone BLE 04136D.

BLE 04136C was prepared in accordance with method D using 2furaldehyde (0.449 mL, 5.42 mmol), KOH (0.276 mg, 4.92 mmol) in methanol (5 mL) and 2-isocyano-l-(pyrrolidin-l-yl)ethanone BLE 04098 (0.75 g, 5.43 mmol). After work-up the residue was purified by column chromatography on silica (cyclohexane:EtOAc = 100:0 to 0:100) to led, after evaporation, to trans(5-(furan-2-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1 -yl)methanone BLE 04136D as a yellow pale oil (0.742 g, 58.5 % yield).

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BLE 04136D

MW: 234.25; Yield: 58.5 %; Yellow Pale Oil.

Ή-NMR (CDC1₃, δ): 1.80-2.10 (m, 4H, CH₂), 3.47-3.60 (m, 3H, CH₂N), 3.944.06 (m, 1H, CH₂N), 4.94 (dd, 1H, J= 7.4 Hz, J= 2.2 Hz, CH-N), 6.14 (d, 1H, J= 7.4 Hz, CH-O), 6.37 (dd, 1H, J= 3.3 Hz,/= 1.8 Hz, CH=C), 6.48 (d, 1H, J = 3.3 Hz, CH=C), 6.93 (d, 1H, J= 2.2 Hz, O-CH=N), 7.44 (d, 1H, J= 1.8 Hz, OCH=C).

¹³C-NMR (CDC1₃, δ): 24.2, 26.0, 46.4, 46.5, 71.3, 74.5, 110.2, 110.5, 143.6,

150.4, 155.0, 166.3.

?raw3ⁱ-(4,5-Dihydro-5-(2-methoxypyridin-3-yl)oxazol-4-yl)(pyrrolidin-lyDmethanone BAL 01014.

BAL 01014 was prepared in accordance with method D using 2-methoxy-

3-pyridinecarboxaldehyde (0.64 ml, 5.43 mmol), KOH (0.305 mg, 5.43 mmol) in methanol (5 mL) and 2-isocyano-l-(pyrrolidin-l-yl)ethanone BLE 04098 (0.75 g, 5.43 mmol). After work-up trans-(4,5-dihydro-5-(2-methoxypyridin-3yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone BAL 01014 was obtained (0.74 mg, 50 % yield) as a white solid.

BAL 01014

MW: 275.30; Yield: 50 %; White Solid; Mp (°C): 110.1.

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R/: 0.25 (EtOAc).

Ή NMR (CDC1₃, δ): 1.82-2.10 (m, 4H, 2xCH₂), 3.40-3.62 (m, 3 H, CH₂N),

3.80-3.90 (m, 3 H, CH₂N), 3.93 (s, 3H, OMe), 4.61 (dd, 1H, J= 7 Hz, J= 2 Hz, CH-N), 6.14 (d, 1H, J= 7 Hz, CH-O), 6.90 (dd, 1H, 7.3 Hz, J= 5 Hz, ArH),

7.02 (d, 1H, 2 Hz, OCH-N), 7.60 (dd, 1H, J= 7.3 Hz, J = 1.7 Hz, ArH)),

8.13 (dd, 1H,, J= 5 Hz, J= 1.8 Hz, ArH).

¹³C-NMR (CDC1₃, δ): 24.3, 26.1, 46.3,46.6, 53.5, 73.5, 78.1, 116.8, 122.2,

135.2, 146.5, 155.3, 160.5 and 167.4.

MS-ESI m/z (% rel. Int.): 276.1 ([MH]⁺, 42).

HPLC: Method A, detection UV 254 nm, BAL 01014 RT = 3.63 min, peak area

97.2 %.

fran5^,-W,W-Diethyl-4,5-dihydro-5-(pyridin-4-yl)oxazole-4-carboxamide SLA 07194A.

SLA 07194A was prepared in accordance with method D using pyridine-

4-carbaldehyde (1.14 mL, 9.52 mmol), KOH (0.54 g, 9.60 mmol) in methanol (5 mL) and7V,7V-diethyl-2-isocyanoacetamide SLA 07184A (1.21 g, 8.65 mmol). After work-up and column chromatography on florisil (ethyl acetate) Zraws-MA’-diethyl-4,5-dihydro-5-(pyridin-4-yl)oxazole-4-carboxamide SLA 07194A was obtained as a brown oil (0.25 g, 12 % yield).

(+/-)

SLA 07194 A

MW: 247.29; Yield: 12 %; Brown Oil.

Ry: 0.15 (AcOEt= 100).

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2018203022 01 May 2018 ’H-NMR (CDC1₃, δ): 1.16-1.34 (m, 6H, CH₃), 3.30-3.80 (m, 4H, CH₂N), 4.60 (dd, 1H, J= 7.7 Hz, J= 2.2 Hz, CH-N), 6.22 (d, 1H, 7.7 Hz, CH-O), 7.06 (d, J= 2.2 Hz, CH=N), 7.23 (d, 2H, J= 5.8 Hz, ArH), 8.61 (d, 2H, 6.0 Hz,

ArH).

Preparation of 2-chloropyridine-4-carbaldehyde SLA 07156.

Methyl 2-chloropyridine-4-carboxylate SLA 07150,

2-Chloro-isonicotinic acid (5.10 g, 32.38 mmol) was dissolved in methanol (150 mL). Thionyl chloride (12 mL) was added. This suspension was stirred 5 h at 70 °C and concentrated in vacuo. The residue was dissolved in dichloromethane (250 mL) washed with a solution of 10 % aqueous K₂CO₃ (2 x 150 mL) dried with MgSC>4, filtered and evaporated. Methyl 2-chloropyridine-

4-carboxylate SLA 07150 was obtained as a yellow solid (5.06 g, 91 %).

SLA 07150

MW: 171.58; Yield: 91 %; Yellow Solid; Mp (°C): 33.0.

R_z: 0.80 (MeOH:CH₂Cl₂= 10:90).

Ή-NMR (CDC1₃, δ): 3.98 (s, 3H, CH₃), 7.78 (dd, 1H, J= 5.1 Hz, J= 1.3 Hz, ArH), 7.89 (d, 1H, J= 0.6 Hz ArH), 8.55 (dd, 1H, J= 5.1 Hz, J= 0.6 Hz, ArH).

(2-Chloropyridin-4-yl)methanol SLA 07152.

Methyl 2-chloropyridine-4-carboxylate (2.50 g, 14.60 mmol) was dissolved in anhydrous THF (50 mL) and this solution was cooled to -78 °C

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2018203022 01 May 2018 under N₂ atmosphere. Diisobutylaluminium hydride 1.0 M in hexanes (63.3 mL, 63.30 mmol) was added dropwise stabilizing the temperature between -50 °C and -70 °C. The reaction mixture was stirred 1.5 h at -78 °C and allowed to stand at room temperature for 3 h. A solution of aqueous 10 % NH₄C1 was slowly added and the mixture was extracted with ethyl acetate (3 x 300 mL). The combined organic layers were washed with water (3 x 20 mL), brine (2 x 20 mL), dried over MgSO₄, filtered and evaporated. (2-Chloropyridin-4yl)methanol SLA 07152 was obtained as a yellow oil (1.97 g, 94 % yield).

HO.

N ci

SLA 07152

MW: 143.71; Yield: 94 %; Yellow Oil.

Ry·: 0.35 (EtOAc:cyclohexane = 30:70).

Ή-NMR (CDCI3, δ): 2.95 (s broad, 1H, OH), 4.75 (s, 2H, CH₂O), 7.21 (dd, 1H, J= 5.1 Hz, J= 1.2 Hz, ArH), 7.37 (d, 1H, J= 1.2 Hz ArH), 8.29 (d, 1H, J= 5.1 Hz, ArH).

MS-ESI m/z (rel. int.): 144.0 ([MH]⁺, 100).

HP LC: Method A, detection UV 254 nm, SLA 07152 RT = 3.45 min, peak area

99.9 %.

2-Chloropyridine-4-carbaldehyde SLA 07156.

In a 250 mL tricol equipped with a low temperature thermometer and two equalizing dropping funnels was charged oxalyl dichloride (1.24 g, 9.81 mmol)

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2018203022 01 May 2018 in dichloromethane (15 mL) and this solution was stirred under N₂ at -78 °C. The first equalizing dropping funnel was connected to a nitrogen flow line and was charged with a solution of (2-chloropyridin-4-yl)methanol SLA 07152 (0.94 g, 6.54 mmol) with dichloromethane (15 mL). The other was charged with a solution of dimethyl sulfoxide anhydrous (1.7 mL, 19.63 mmol) in dichloromethane (2 mL) and this solution was added dropwise (25 min) in order to stabilize the temperature between -60 °C and -70 °C. At the end of the addition the reaction solution was warmed to -60 °C over a period of 20 min then the solution of (2-chloropyridin-4-yl)methanol SLA 07152 was added dropwise (50 min) keeping the temperature between -50 °C and -60 °C in the reactor then the mixture reaction was warmed to -45 °C over a period of 30 min. The dropping funnel was washed with dichloromethane (2x5 mL) and charged with a solution of triethylamine (480 μΐ, 6.51 mmol) in dichloromethane (4 mL) which was added (10 min) to the reaction mixture and finally the reaction flask was allowed to warm to 0 °C over 10 min. The reaction solution was transferred to a 500 mL separatory funnel charged with 130 mL of a 5 % aqueous NH₄C1 solution. The two phases were separated the aqueous phase was extracted with dichloromethane (3 x 50 mL) and the combined organic phases were washed with 1 M aqueous phosphate buffer (pH = 7; 4xl00mL), then dried over MgSO₄, filtered and evaporated. 2-Chloropyridine-4-carbaldehyde SLA 07156 was obtained as an orange solid (0.740 g, 76 % yield).

.0

N Cl

SLA 07156

MW: 141.57; Yield: 76 %; Orange Solid.

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Ry·: 0.35 (EtOAc:cyclohexane = 30:70).

^-NMR (CDC1₃, δ): 7.65 (dd, 1H, J= 5.0 Hz, J= 1.3 Hz, ArH), 7.75 (d, 1H, J = 1.3 Hz, ArH) 8.66 (d, 1H, J= 5.0 Hz, ArH), 10.05 (s, 1H, CHO).

/raw5^,-(5-(2-Chloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-lvDmethanone SLA 07174.

SLA 07174 was prepared in accordance with method D using 2chloropyridine-4-carbaldehyde SLA 07156 (0.12 g, 1.05 mmol), KOH (0.06 g, 1.05 mmol) in methanol (10 mL) and 2-isocyano-l-(pyrrolidin-l-yl)ethanone BLE 04098 (0.146 g, 1.05 mmol). The solution was stirred 24 h with continued cooling. After work-up /ra7w-(5-(2-chloropyridin-4-yl)-4,5-dihydrooxazol-4yl)(pyrrolidin-l-yl)methanone SLA 07174 was obtained as a yellow solid (0.19 g, 66 % yield).

(+/-)

SLA 07174

MW: 279.72; Yield: 66 %; Yellow Solid; Mp (°C): 116.3.

Ή-NMR (CDC1₃, δ): 1.86-2.07 (m, 4H, CH₂), 3.45-3.62 (m, 3H, CH₂N), 3.934.01 (m, 1H, CH₂N), 4.50 (dd, J= 8.0 Hz, J= 2.3 Hz, 1H, CH-N), 6.19 ( d, 1H, 7= 8.0 Hz, CH-O), 7.02 (d, 1H, .7=2,3 Hz, CH=N), 7.17 (td, 1H, 7=5.1 Hz 7= 0.9 Hz, 7= 0.4 Hz, ArH), 7.29 (d, 1H, 7= 0.7 Hz, ArH), 8.38 (d, 1H, 7= 5.1 Hz, ArH).

¹³C-NMR (CDC1₃, δ): 22.5,24.4,44.9,45.0, 74.3, 77.3,117.2, 119.0, 148.6,

150.4, 150.6, 153.1, 164.0.

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2018203022 01 May 2018 ?raHs-(5-(3-Bromopyridin-4-yl)-4,5-dihydrooxazol-4-yl)(pyiTolidni-lypmethanone BAL 01028A.

BAL 01028A was prepared in accordance with method D using 3-bromo4-pyridinecarboxaldehyde (1.010 g, 5.43 mmol), KOH (0.305 g, 5.43 mmol) in methanol (5 mL) and 2-isocyano-l-pyrrolidin-l-yl-ethanone BLE 04134 (0.75 g, 5.43 mmol). The mixture was stirred at 0 °C until precipitation and concentrated. The mixture was partitioned between EtOAc (50 ml) and H₂O (25 ml). The aqueous layer was extracted twice with EtOAc (25 mL). The EtOAc fractions were combined, washed twice with brine (2x25 mL), dried over MgSO4 and filtered. After evaporation and fi7Z72s-(5-(3-bromopyridin-4-yl)-4,5dihydrooxazol-4-yl)(pyrrolidin-l-yl)methanone BAL 01028A was obtained (1.20 g, 68 % yield) as a white solid.

(+/-)

BAL 01028A

MW: 324.17; Yield: 68 %; White Solid; Mp (°C): 160.8.

R/’: 0.25 (EtOAc = 100).

Ή NMR (CDC1₃, δ): 1.82-2.08 (m, 4H, 2xCH₂), 3.45-3.65 (m, 3H, CH₂N),

3.80-3.92 (m, 1H, CH₂N), 4.60 (dd, 1H, J= 2.1 Hz, J= 6.1 Hz, CH-N), 6.30 (d, 1H, J= 6.1 Hz, CH-O), 7.10 (d, 1H, J= 2.1 Hz, OCH=N), 7.30(d, 1H, J= 5.0 Hz, ArH)), 8.55 (d, 1H, J= 5.0 Hz, ArH), 8.72 (s, 1H, ArH).

¹³C-NMR (CDC1₃, δ): 24.3, 26.0,46.4,46.6, 74.5, 79.6, 118.6, 121.1, 148.3,

148.8, 152.1, 155.1, 166.2.

MS-ESI m/z (% rel. Int.): 324.1/326.1 ([MH]⁺, 50/50), 239.0 (100).

HPLC: Method A, detection UV 254 nm, BAL 01028A RT = 3.50 min, peak area 96.8 %.

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2018203022 01 May 2018 fmn5^,-(5-(3-Chloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-lvDmethanone BAL 01028B.

BAL 01028B was prepared in accordance with method D using 2isocyano-l-pyrrolidin-l-yl-ethanone BLE 04134 (0.75 g, 5.43 mmol), KOH (0.305 g, 5.43 mmol) in methanol (5 mL) and 3-chloro-isonicotinaldehyde (0.769 g, 5.43 mmol). The solution was stirred 3 h at 0 °C. trans-(5-(3Chloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1 -yl)methanone BAL 01028B (1.20 g, 65% yield) was obtained as a white solid.

(+/-)

BAL 01028B

MW: 279.72; Yield: 65 %; White Solid; Mp (°C): 162.

R/: 0.25 (EtOAc = 100).

’H NMR (CDC1₃, δ): 1.82-2.08 (m, 4H, CH₂), 3.45-3.65 (m, 3H, CH₂N), 3.823.93 (m, 1H, CH₂N), 4.62 (dd, 1H, J= 2.1 Hz, J= 6.1 Hz, CH-N), 6.38 (d, 1H, J= 6.1 Hz, CH-O), 7.08 (d, 1H, J= 2.1 Hz, OCH=N), 7.33 (d, 1H, 5.0 Hz,

ArH), 8.52 (d, 1H, J= 5.0 Hz, ArH), 8.59 (s, 1H, ArH).

¹³C-NMR (CD₃OD, δ): 24.3, 26.0, 46.4,46.6, 74.4, 77.9,120.6, 128.8, 146.6,

148.3, 149.7, 155.0, 166.1.

MS-ESI m/z (% rel. Int.): 280.1/282.1 ([MH]⁺, 39/14).

HPLC: Method A, detection UV 254 nm, BAL 01028B RT = 3.47 min, peak area 97.2 %.

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2018203022 01 May 2018 /mH5^,-(5-(2-Chloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(2.fi^r-pyrrol-l(5Z/)yljmethanone SLA 07158.

SLA 07158 was prepared in accordance with method D using 2chloropyridine-4-carbaldehyde SLA 07156 (0.47 g, 3.31 mmol), KOH (0.184 g,

3.33 mmol) in methanol (10 mL) and 2-Isocyano-l-(277-pyrrol-1(577)yl)ethanone SLA 07178 (0.410 g, 3.01 mmol). The solution was stirred 2 h with continued cooling. After work-up and column chromatography on florisil (EtOAc), ira«5-(5-(2-chloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(277-pyrroll(577)-yl)methanone SLA 07158 was obtained as a yellow solid (0.597 g, 84 %)·

SLA 07158

MW: 277.71; Yield: 84 %; Yellow Solid; Mp (°C): 90.2.

R_z: 0.10 (EtOAc).

Ή-NMR (CDC1₃, δ): 4.26-4.37 (m, 3H, CH₂N), 4.48-4.52 (dd, 1H, J= 2.3 Hz, J = 8.0 Hz, CH-N), 4.75-4.85 (m, 1H, CH₂N), 5.80-5.95 (m, 2H, CH=CH), 6.20 (d, 1H, J= 8 Hz, CH-O), 7.02 (d, J= 2.3 Hz, CH=N), 7.17 (td, 1H, J= 5.1 Hz, J = 0.8 Hz, J- 0.6 Hz, ArH), 7.30 (t, 1H, J= 0.6 Hz, ArH), 8.38 (d, 1H, J= 5.1 Hz, ArH).

¹³C-NMR (CDC1₃, δ): 53.4, 53.9, 75.8, 78.9, 118.9, 120.7, 125.2, 125.4, 150.3,

151.9, 152.3, 154.

fraHs-(4,5-Dihydro-5-(pyridin-4-yl)oxazol-4-yl)(277-pyrrol-l(577)-yl)methanone SLA 07180.

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SLA 07158 was prepared in accordance with method D using pyridine-4carbaldehyde (0.293 mL, 2.40 mmol), KOH (0.13 g, 2.32 mmol) in methanol (10 mL) and 2-isocyano-l-(2ZZ-pyrrol-l(5Z/)-yl)ethanone SLA 07178 (0.301 g,

2.20 mmol). The solution was stirred 2 h with continued cooling. After work-up and column chromatography on florisil (EtOAc), trans-(A,5-dihydro-5-(pyridin4-yl)oxazol-4-yl)(2#-pyrrol-l(5£/)-yl)methanone SLA 07180 was obtained (0.284 g, 53 % yield) as a yellow oil.

(+/-)

SLA 07180

MW: 243.26; Yield: 53 %; Yellow Oil.

R_z: 0.15 (AcOEt).

Ή-NMR (CDC1₃, δ): 4.28-4.33 (m, 3H, CH₂N), 4.52-4.56 (dd, 1H, 7= 7.8 Hz, J = 2.2 Hz, CH-N), 4.73-4.82 (m, 1H, CH₂N), 5.80-5.93(m, 2H, CH=CH), 6.18 (d, 1H, J= 7.8 Hz, CH-O), 7.08 (d, 7= 2.2 Hz, CH=N), 7.27 (d, 2H, 7= 6.0 Hz, ArH), 8.59 (d, 2H, 7= 6.0 Hz, ArH).

¹³C-NMR (CDCI3, δ): 53.6, 53.9, 75.8, 79.7, 120.2, 125.3, 125.6, 148.7, 150.5,

155.3, 166.1.

Preparation of Compound 20, Compound 2 L Compound 22, Compound 23, Compound 24, Compound 25, Compound 26, Compound 27, Compound 28, Compound 29, Compound 30, Compound 31, Compound 32, Compound 34, Compound 35, Compound 36, Compound 37, Compound 38, Compound 39, Compound 40, Compound 41, Compound 42, Compound 43, Compound 44, Compound 45, Compound 46, Compound 48, Compound 49 and Compound 50.

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General method for oxazolines acidic hydrolysis: method E: DL-/Z?re<7-2-Amino-3-hydroxy-3-(pyridin-3-yl)-1 -fpyrrolidin-1 -vDpropan-1 -one dihydrochloride Compound 20.

To a solution of trans-(4,5-dihydro-5-(pyridin-3-yl)oxazol-4yl)(pyrrolidin-l-yl)methanone BLE 04110B (0.932 g, 3.80 mmol) in methanol (10 mL) was added hydrochloric acid 37 % (1.2 mL). After heating (50 °C) the mixture for 2.25 h the reaction mixture was concentrated and the crude product was coevaporated twice with ethyl acetate. After trituration with ethyl acetate, filtration and drying DL-/7ireo-2-amino-3-hydroxy-3-(pyridin-3-yl)-l(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 20 was obtained as a white solid (1.10 g, 94 % yield).

OH o

Compound 20

MW: 308.2; Yield: 94 %; White Solid; Mp (°C): 123.4.

Ή-NMR (CD₃OD, δ): 1.65-2.00 (m, 4H, 2xCH₂), 2.82-3.11 (m, 1H, -CH₂N),

3.30-3.57 (m, 2H, CH₂N), 3.57-3.77 (m, 1H, CH₂N), 4.54 (d, 1H, J= 5.3 Hz, CH-N), 5.38 (d, 1H, J= 5.3 Hz, CH-O), 8.15 (dd, 1H, J= 7.6 Hz, J= 5.0 Hz, ArH), 8.68 (d, 1H, J = 7.6 Hz, ArH), 8.89 (d, 1H, J= 7.6 Hz, ArH), 8.96 (s, 1H, ArH).

¹³C-NMR (CD₃OD, δ): 24.9, 26.9, 47.7,48.2, 58.1, 69.6, 128.7, 141.5, 141.6,

143.1, 146.5, 165.4.

DL-^rgo-2-Amino-3-hydroxy-3-(pyridin-2-yl)-1 -(pyrrolidin-1 -yDpropan-1 -one dihydrochloride Compound 21.

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Compound 21 was prepared following method E with //7^-(4,5-dihydro-

5-(pyridin-2-yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone BLE 04110 B (0.44 g, 1.79 mmol), hydrochloric acid 37 % (1.0 mL) and methanol (10 mL). After 2.5 h at 50 °C and work-up DL-i7/reo-2-amino-3-hydroxy-3-(pyridin-2-yl)-l(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 21 was obtained as a yellow solid (0.44 g, 84 % yield).

OH 0 (+/-) nh₂

HCI

Compound 21

MW: 308.28; Yield: 84%; Yellow Solid.

’H-NMR (CD₃OD, δ): 1.75-2.01 (m, 4H, 2xCH₂), 3.10-3.22 (m, 1H, CH₂N),

3.39-3.60 (m, 2H, CH₂N), 3.63-3.75 (m, 1H, CH₂N), 4.71 (d, 1H, J = 5.0 Hz, CH₂N), 5.55 (d, 1H, J= 5.0 Hz, CH-O), 8.05 (t, 1H, J= 6.4 Hz, ArH), 8.13 (d, 1H, J= 8.0 Hz, ArH), 8.61 (t, 1H, 8.0 Hz, ArH), 8.84 (d, 1H, J = 5.6 Hz,

ArH).

DL-/Arg<?-2-Amino-3-hydroxy-3-(pyridin-4-yl)-1 -(pyrrolidin-1 -yPpropan-1 -one dihydrochloride Compound 22.

Compound 22 was prepared following method E with iraws^,-(4,5-dihydro-

5-(pyridin-4-yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone Compound 19 (0.750 g, 3.07 mmol), hydrochloric acid 37 % (1.0 mL) and methanol (10 mL). After 3.0 h at 50 °C and work-up DL-^reo-2-amino-3-hydroxy-3-(pyridin-4-yl)-l(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 22 was obtained as a white solid (0.935 g, 99 % yield).

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Compound 22

MW: 308.28; Yield: 99 %; White Solid; Mp (°C): 117.0.

^-NMR (CD₃OD, δ): 1.75-2.03 (m, 4H, 2xCH₂), 2.93-3.08 (m, 1H, CHN), 3.32-3.75 (m, 3H, 2xCH₂), 4.54 (d, 1H, J= 5.9 Hz, CH-N), 5.40 (d, 1H, J= 5.9 Hz, CH-O), 8.21 (d, 2H, J= 5.8 Hz, ArH), 8.94 (d, 2H, J= 5.8 Hz, ArH). MS-ESI m/z (% rel. int.): 236.1 ([MH]+, 17), 219 (25), 148 (100).

HPLC: Method A, detection UV 254 nm, Compound 22 RT = 0.8 min, peak area 96.3 %.

DL-/Areo-2-Amino-3 -hydroxy-1 -(pyrrolidin-1 -yl)-3-(thiophen-3-yl)propan-1 one hydrochloride Compound 23.

Compound 23 was prepared following method E with /ra«5-(4,5-dihydro5-(thiophen-3-yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone BLE 04124A (0.486 g, 1.94 mmol), hydrochloric acid 37 % (0.6 niL) and methanol (10 mL). After

3.5 h at 50 °C and work-up DL-/Areo-2-amino-3-hydroxy-l-(pyrrolidin-l-yl)-3(thiophen-3-yl)propan-l-one hydrochloride Compound 23 was obtained as a white solid (0.480 g, 89.5 % yield).

(+/-)

Compound 23

MW: 276.7; Yield: 89.5 %; White Solid; Mp (°C): 227.4.

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Ή-NMR (CD₃OD, δ): 1.47-1.88 (m, 4H, 2xCH₂), 2.31-2.46 (m, 1H, CH₂N), 3.18-3.46 (m, 3H, CH₂N), 4.16 (d, 1H, J= 9.0 Hz, CH-N), 4.97 (d, 1H, J= 9.0 Hz, CH-O), 7.14 (dd, 1H, J= 4.9 Hz, J= 1.1 Hz, ArH), 7.40-7.50 (m, 2H, ArH).

¹³C-NMR (CD₃OD, δ): 24.9, 26.7, 47.3, 47.6, 59.2, 70.6, 124.1, 127.1, 127.7,

142.3, 166.3.

DL-Z/zreo-2-Amino-3 -hydroxy-1 -(pyrrolidin-1 - yl)-3-(thiophen-2-yl)propan-1 one hydrochloride Compound 24.

Compound 24 was prepared following method E with trans-(A, 5-dihydro5-(thiophen-2-yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone BLE 04124B (0.677 g, 2.70 mmol), hydrochloric acid 37 % (0.6 mL) and methanol (10 mL). After

3.5 h at 50 °C and work-up DL-/Areo-2-amino-3-hydroxy-l-(pyrrolidin-l-yl)-3(thiophen-2-yl)propan-l-one hydrochloride Compound 24 was obtained as a white solid (0.630 g, 84.5 % yield).

Compound 24

MW: 276.7; Yield: 84.5 %; White Solid; Mp (°C): 183.2.

Ή-NMR (CD₃OD, δ): 1.49-1.90 (m, 4H, 2xCH₂), 2.36-2.48 (m, 1H, CH₂N),

3.20-3.48 (m, 3H, CH₂N), 4.18 (d, 1H, J= 9.1 Hz, CH-N), 5.14 (d, 1H, J= 9.1 Hz, CH-O), 7.00-7.08 (m, 2H, ArH), 7.45 (dd, 1H, J= 4.9 Hz, J= 1.6 Hz, ArH). ¹³C-NMR (CD3OD, δ): 24.9, 26.8,47.3,47.7, 59.6, 70.5, 126.3, 127.0, 128.2,

144.5, 166.1.

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DL-/Areo-2-amino-3 -hydroxy-1 -(pyrrolidin-1 -yl)-3 -(thiazol-2-yl)propan-1 -one dihydrochloride Compound 25.

Compound 25 was prepared following method E with trans-(4,5-dihydro5-(thiazol-2-yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone BLE 04124C (0.558 g, 2.22 mmol), hydrochloric acid 37 % (0.6 mL) and methanol (10 mL). After 3.5 h at 50 °C and work-up DL-/Areo-2-amino-3-hydroxy-l-(pyrrolidin-l-yl)-3(thiazol-2-yl)propan-l-one dihydrochloride Compound 25 was obtained as a pale yellow solid (0.532 g, 76.5 % yield).

2.HCI (+/-)

Compound 25

MW: 276.7; Yield; 76.5 %; Pale Yellow Solid; Mp (°C): 145.8.

Ή-NMR (CD₃OD, δ): 1.75-2.00 (m, 4H, 2xCH₂), 3.05-3.17 (m, 1H, -CH₂N), 3.36-3.58 (m, 2H, CH₂N), 3.58-3.70 (m, 1H, CH₂N), 4.67 (d, 1H, J= 5.4 Hz, CH-N), 5.49 (d, 1H, J= 5.4 Hz, CH-O), 7.84 (d, 1H, J= 3.4 Hz ArH), 7.99 (d, 1H, 7=3.4 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 24.9, 27.0, 47.7,48.0, 57.5, 69.9,123.6, 142.1,165.3,

173.3.

DL-^reo-2-Amino-3-(3a,7a-dihydrobenzorfe]thiophen-3-yl)-3-hydroxy-l(pyrrolidin-l-yl)propan-l-one hydrochloride Compound 26.

Compound 26 was prepared following method E with fo6w-(5(benzo[b]thiophen-3-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-l-yl)methanone BLE 04124D (1.050 g, 3.49 mmol), hydrochloric acid 37 % (1.2 mL) and

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2018203022 01 May 2018 methanol (10 mL). After 3.5 h at 50 °C and work-up DL-Z/zreo-2-amino-3(3a,7a-dihydrobenzo[&]thiophen-3-yl)-3-hydroxy-1 -(pyrrolidin-1 -yl)propan-1one hydrochloride Compound 26 was obtained as a white solid (0.970 g, 85 % yield).

S J h₂N

2.HCI (+/-)

Compound 26

MW: 326.84; Yield: 85 %; White Solid; Mp (°C): 207.0.

’H-NMR (CD₃OD, δ): 0.92-1.09 (m, 2H, 2xCH₂), 1.42-1.60 (m, 2H, 2xCH₂), 1.83-1.98 (m, 1H, CH₂N), 2.76-2.91 (m, 1H, CH₂N), 3.06-3.25 (m, 2H, CH2N), 4.30 (d, 1H, J= 9.5 Hz, CH-N), 5.29 (d, 1H, 9.5 Hz, CH-O), 7.35-

7.43 (m, 2H, ArH), 7.78-7.89 (m, 2H, ArH), 7.90-7.97 (m, 1H, ArH). ¹³C-NMR (CD₃OD, δ): 24.5, 26.4, 47.3,47.4, 59.0, 69.5, 123.1, 124.0, 125.4,

126.1, 126.8, 136.6, 138.3, 141.9,166.1.

DL-/Areo-2-Amino-3-(ftiran-3-yl)-3-hydroxy-1 -(pyrrolidin-1 -yPpropan-1 -one hydrochloride Compound 27.

Compound 27 was prepared following method E with /ra«5-(5-(furan-3yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-l-yl)methanone BLE 04 BOB (0.800 g,

3.41 mmol), hydrochloric acid 37 % (0.6 mL) and methanol (10 mL). After 3.5 h at 50 °C and work-up DL-/Zereo-2-amino-3-(furan-3-yl)-3-hydroxy-l(pyrrolidin-l-yl)propan-l-one hydrochloride Compound 27 was obtained as a white solid (0.738 g, 83 % yield).

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(+/-)

Compound 27

MW: 260.72; Yield: 83 %; White Solid; Mp (°C): 218.0.

Ή-NMR (CD₃OD, δ): 1.62-1.95 (m, 4H, 2xCH₂), 2.82-2.95 (m, 1H, CH₂N),

3.22-3.38 (m, 1H, CH₂N), 3.39-3.55 (m, 2H, CH₂N), 4.19 (d, 1H, J= 8.4 Hz, CH-N), 4.90 (d, 1H, J= 8.4 Hz, CH-O), 6.49 (m, 1H, ArH), 7.52-7.57 (m, 2H, ArH).

¹³C-NMR (CD₃OD, δ): 24.9, 26.7, 47.4, 48.0, 58.7, 67.2, 109.8, 125.9, 142.0,

145.2, 166.3.

DL-//zreo-2-Amino-3-hydroxy-3-(naphthalen-2-yl)- l-(pyrrolidin- l-yl)propan-1one hydrochloride Compound 28.

Compound 28 was prepared following method E with /ra7zs-(4,5-dihydro5-(naphthalen-3-yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone BLE 04130C (0.745 g, 2.53 mmol), hydrochloric acid 37 % (0.6 mL) and methanol (10 mL). After

3.5 h at 50 °C and work-up DL-/Areo-2-amino-3-hydroxy-3-(naphthalen-2-yl)- l-(pyrrolidin-l-yl)propan-l-one hydrochloride Compound 28 was obtained as a white solid (0.706 g, 87 % yield).

Compound 28

MW: 320.81; Yield: 87 %; White Solid; Mp (°C): 173.8.

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2018203022 01 May 2018 ^-NMR (CD₃OD, δ): 0.93-1.10 (m, 1H, CH₂), 1.20-1.37 (m, 1H, CH₂), 1.44-

1.71 (m, 2H, CH₂), 1.99-2.10 (m, 1H, CH₂N), 3.11-3.26 (m, 2H, CH2N), 3.31-

3.41 (m, 1H, CH2N), 4.23 (d, 1H, J= 9.1 Hz, CH-N), 5.06 (d, 1H, J= 9.1Hz, CH-O), 7.50-7.63 (m, 3H, ArH), 7.87-7.97 (m, 4H, ArH).

¹³C-NMR (CD₃OD, δ): 24.6, 26.3, 47.2,47.5, 59.4, 74.3, 125.1, 126.9, 127.7,

127.8,128.8, 129.0, 129.4, 134.5, 135.0, 138.0, 166.4.

DL-/7/reo-2-Amino-3 -hydroxy-3-(naphthalen-1 -yl)-1 -(pyrrolidin-1 -vDpropan-1 one hydrochloride Compound 29.

Compound 29 was prepared following method E with Zmi5-(4,5-dihydro5-(naphthalen-4-yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone BLE 04130D (0.794 g, 2.69 mmol), hydrochloric acid 37 % (0.6 mL) and methanol (10 mL). After

3.5 h at 50 °C and work-up DL-i/zreo-2-amino-3-hydroxy-3-(naphthalen-l-yl)-

1-(pyrrolidin-l-yl)propan-l-one hydrochloride Compound 29 was obtained as a white solid (0.768 g, 89 % yield).

Compound 29

MW: 320.81; Yield: 89 %; White Solid; Mp (°C): 177.8.

Ή-NMR (CD₃OD, δ): 0.71-0.91 (m, 2H, CH₂), 1.29-1.51 (m, 3H, CH₂), 2.54-

2.67 (m, 1H, CH₂N), 2.88-3.02 (m, 1H, CH₂N), 3.02-3.16 (m, 1H, CH₂N), 4.27 (d, 1H, J= 9.8 Hz, CH-N), 5.67 (d, 1H, J= 9.8 Hz, CH-O), 7.50-7.61 (m, 3H, ArH), 7.90-7.98 (m, 3H, ArH), 8.08-8.14 (m, 1H, ArH).

¹³C-NMR (CD₃OD, δ): 24.4, 26.2, 47.1,47.3, 59.5, 70.3, 124.0, 126.5 (2xC),

127.2,127.4,129.9, 130.4, 132.1, 135.0,137.1, 166.1.

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DL-^reo-2-Amino-3-hydroxy-1 -(pyrrolidin-1 -yl)-3 -(quinolin-2-yl)propan-1 one dihydrochloride Compound 30.

Compound 30 was prepared following method E with Zraws-(4,5-dihydro5-(quinolin-2-yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone BLE 04136B (0.923 g,

3.13 mmol), hydrochloric acid 37 % (0.6 mL) and methanol (15 mL). After 3.5 h at 50 °C and work-up DL-/Areo-2-amino-3-hydroxy-1-(pyrrolidin-l-yl)-3(quinolin-2-yl)propan-l-one dihydrochloride Compound 30 was obtained as a yellow solid (1.098 g, 98 % yield).

Compound 30

MW: 358.26; Yield: 98 %; Yellow Solid; Mp (°C): 131.5.

Ή-NMR (CD₃OD, δ): 1.69-2.07 (m, 4H, CH₂), 3.16-3.34 (m, 3H, CH₂), 3.373.60 (m, 2H, CH₂N), 3.77-3.88 (m, 1H, CH₂-N), 5.85 (d, 1H, 4.9 Hz, CHO), 8.03 (t, 1H, J= 7.6 Hz, ArH), 8.17-8.30 (m, 2H, ArH), 8.40 (d, 1H, J= 8.3 Hz, ArH), 8.56 (d, 1H, J= 8.6 Hz, ArH), 9.25 (d, 1H, J= 8.6 Hz, ArH), not seen under H₂O (d, 1H, CH-NH₂).

¹³C-NMR (CD₃OD, δ): 24.9,27.0, 47.9,48.2, 57.3, 70.3,121.5, 122.5,130.4,

130.5,131.5, 136.5, 140.2, 148.5,157.8,164.8.

DL-f/rrgo-2-Amino-3 -hydroxy-3 -(isoquinolin-4-yl)-1 -(pyrrolidin-1 -yl)propan-1 one dihydrochloride Compound 31.

Compound 31 was prepared following method E with trans-(f,5-dihydro5-(isoquinolin-4-yl)oxazol-4-yl)(pyrrolidin-1 -yl)methanone BLE 04136C

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Compound 31

MW: 358.27; Yield: 99 %; Off White Solid; Mp (°C): 158.5.

Ή-NMR (CD₃OD, δ): 1.04-1.32 (m, 2H, CH₂), 1.51-1.72 (m, 2H, CH₂), 2.05-

2.20 (m, 1H, CH₂N), 2.68-2.80 (m, 1H, CH₂N), 3.20-2.47 (m, 1H, CH2N), 4.57 (d, 1H, J= 8.5 Hz, CH-NH₂), 5.99 (d, 1H, J= 8.5 Hz, CH-OH), 8.09 (t, 1H, J=

8.6 Hz, ArH), 8.27 (t, 1H, J= 8.6 Hz, ArH), 8.38 (d, 1H, J= 8.6 Hz, ArH), 8.45 (d, 1H, J= 8.6 Hz, ArH), 8.55 (d, 1H, 5.6 Hz, ArH), 9.35 (d, 1H, J= 5.6 Hz,

ArH).

¹³C-NMR (CD₃OD, δ): 24.6, 26.5, 47.5,48.0, 58.2, 69.1,122.3, 122.6, 125.9,

127.7, 131.7, 136.7, 138.9, 146.1, 159.4, 165.2.

W-(DL-/Areo-l-hydroxy-3-oxo-3-(pyrrolidin-l-yl)-l-(quinolin-3-yl)propan-2ypformamide hydrochloride Compound 32.

Compound 32 was prepared following method E with /ra//5-(4,5-dihydro5-(quinolin-3-yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone BAL 01016 (0.905 g,

3.41 mmol), hydrochloric acid 37 % (0.6 mL) and methanol (10 mL). After 2 h at RT and work-up 7V-(DL-i/zreo-l-hydroxy-3-oxo-3-(pyrrolidin-l-yl)-l(quinolin-3-yl)propan-2-yl)formamide hydrochloride Compound 32 was obtained as a white solid (240 mg, 20.0 % yield).

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Compound 32

MW: 349.81; Yield: 20.0 %; White Solid; Mp (°C): 203.2.

’H NMR (CD₃OD, δ): 1.78-2.09 (m, 4H, CH₂), 3.35-3.58 (m, 2 H, CH₂N), 3.58-

3.80 (m, 2 H, CH₂N), 5.28 (d, 1H, J= 4 Hz, CH-N), 5.51 (d, 1H, J= 4 Hz, CHO), 8.00 (t, 2H, J= 7.1 Hz, ArH)), 8.18 (t, 1H, J= 6.9 Hz, ArH), 8.26 (d, 1H, J = 8.6 Hz, ArH), 8.36 (d, 1H, J= 8.3 Hz, ArH), 9.18 (s, 1H, CHO), 9.26 (s, 1H, ArH).

¹³C-NMR (CD₃OD, δ): 25.1, 27.0, 47.5,48.3, 55.5, 71.2, 121.4, 129.9, 130.6,

131.5, 136.2, 137.3, 138.5, 145.3, 145.8, 163.4, 168.7.

MS-ESI m/z (% rel. Int.): 314 ([MH]⁺, 50), 158.1 (100).

HPLC: Method A, detection UV 254nm, Compound 32 RT = 3.36 min, peak area 99.9 %.

DL-/7?reo-2-Amino-3-hydroxy-l-(pyrrolidin-l-yl)-3-(quinolin-3-yl)propan-lone dihydrochloride Compound 33.

Compound 33 was prepared following method E with trans-(A, 5-dihydro5-(quinolin-3-yl)oxazol-4-yl)(pyrrolidin-l-yl)methanoneBAL 01016 (0.91 g, 3.41mmol), hydrochloric acid 37% (0.6 mL) and methanol (10 mL). After 3 h at 50 °C and work-up DL-i/zreo-2-amino-3-hydroxy-l-(pyrrolidin-l-yl)-3(quinolin-3-yl)propan-l-one dihydrochloride Compound 33 (678 mg, 55 % yield) was obtained as a white solid.

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Compound 33

MW: 358.26; Yield: 55 %; White Solid; Mp (°C): 190.9.

^!H NMR (CD₃OD, δ): 1.57-1.80 (m, 2H, CH₂), 1.80-1.99 (m, 2H, CH₂), 3.01-

3.20 (m, 1H, CH₂N), 3.35-3.61(m, 2H, CH₂N), 3.61-3.82(m, 1H, CH₂N), 4.70 (d, 1H, J= 5.0 Hz, CH-N), 5.58 (d, 1H, J= 5.0 Hz, CH-O), 7.96-8.11 (m, 1H, ArH), 8.18-8.29 (m, 1H, ArH), 8.29-8.38 (m, 1H, ArH), 8.38-8.49 (m, 1H, ArH), 9.28 (s, 1H, ArH), 9.34 (s, 1H, ArH).

¹³C-NMR (CD₃OD, δ): 24.9,26.9,47.8,48.3, 58.2, 69.8, 121.8, 130.0,130.8,

131.9,135.4, 136.9, 139.3,145.1, 146.2, 165.6.

MS-ESI m/z (% rel. Int.): 286.2 ([MH]⁺,100).

HPLC: Method A, detection UV 254 nm, Compound 33 RT = 3.15 min, peak area 97.0 %.

DL-/Areo-2-Amino-3 -(2-chloropyridin-4-yl)-3 -hydroxy-1-(277-pyrrol-1 (577)yl)propan-l-one dihydrochloride Compound 34.

Compound 34 was prepared following method E with /ran5-(5-(2chloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(277-pyrrol-l(577)-yl)methanone SLA 07158 (0.597 g, 2.02 mmol), hydrochloric acid 37 % (1.0 mL) and methanol (10 mL). After 2 h at room temperature and work-up DL-/Areo-2amino-3-(2-chloropyridin-4-yl)-3-hydroxy-1 -(277-pyrrol-1 (577)-yl)propan-1 -one dihydrochloride Compound 34 (0.656 mg, 91 % yield) was obtained as a pale yellow solid.

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OH 0

V N \

NH₂

HCI

Compound 34

MW: 340.63; Yield: 91 %; Pale Yellow Solid; Mp (°C): 196.2.

^-NMR (CD₃OD, δ): 3.45-3.50 (m, 1H, CH2N), 4.04-4.15 (m, 1H, CH₂N),

4.22-4.36 (m, 3H, CH₂N & CHNH₂), 5.05 (d, 1H, J= 7.1 Hz, -CHO), 5.71 (d, 1H, J= 4.3 Hz, CH=CH), 5.84 (d, 1H, J= 4.3 Hz, CH=CH), 7.47 (d, 1H, 7 = 5.0 Hz, ArH), 7.57 (s, 1H, ArH), 8.39 (d, 1H, 7= 5.0 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 54.3, 54.5, 58.2, 71.7, 122.0, 123.5, 125.7, 126.3, 151.0, 152.8,154.0, 165.8.

DL-/7zreo-2-Amino-3-hydroxy-3-(pvridin-4~yl)-1 -fpiperidin-1 -yl)propan-1 -one dihydrochloride Compound 35.

Compound 35 was prepared following method E with trans-(A,5-dihydro5-(pyridin-4-yl)oxazol-4-yl)(piperidin-l-yl)methanone SLA 07122A (0.33 g,

1.27 mmol), hydrochloric acid 37 % (1.0 mL) and methanol (10 mL). After 3 h at 50 °C and work-up DL-i/zreo-2-amino-3-hydroxy-3-(pyridin-4-yl)-l(piperidin-l-yl)propan-l-one dihydrochloride Compound 35 was obtained as a yellow solid (0.375 g, 91 % yield).

OH O

II I £ '

NH₂ (+/.) 2. HCI

Compound 35

MW: 322.31; Yield: 91 %; Yellow Solid; Mp (°C): 145.

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2018203022 01 May 2018 ^-NMR (CD₃OD, δ): 1.05-1.17 (m, 1H, CH₂) 1.28-1.65 (m, 5H, CH₂), 2.753.00 (m, 1H, CH₂N), 3.10-3.22 (m, 1H, CH₂N), 3.23-3.38 (m, 1H, CH₂N), 3.533.65 (m, 1H, CH₂N), 4.68 (d, 1H, J= 5.8 Hz, CHNH₂), 5.14 (d, 1H, J= 5.8 Hz, CHO), 8.06 (d, 2H, J= 6.0 Hz, ArH), 8.79 (d, 2H, J= 6.5 Hz, ArH).

DL-rAreo-2-Amino-3-hydroxy-3-(pyridin-4-yl)-1 -(morpholin-1 -yDpropan-1 -one dihydrochloride Compound 36.

Compound 36 was prepared following method E with Zra«5-(4,5-dihydro5-(pyridin-4-yl)oxazol-4-yl)(morpholino)methanone SLA 07124A (0.146 g, 0.56 mmol), hydrochloric acid 37 % (1.0 mL) and methanol (10 mL). After 3 h at 50 °C and work-up DL-/Areo-2-amino-3-hydroxy-3-(pyridin-4-yl)-l(morpholin-l-yl)propan-l-one dihydrochloride Compound 36 was obtained as a pale yellow solid (0.143 g, 89 % yield).

OH o

(+/.) 2 HCI

Compound 36

MW: 287.78; Yield: 89 %; Pale Yellow Solid; Mp (°C): 115.9.

’H-NMR (CD₃OD, δ): 3.32-3.82 (m, 8H, 4xCH₂), 5.41 (d, 1H, J= 5.0 Hz, CHO-), 8.28 (d, 2H, J= 5.9 Hz, ArH), 8.97 (d, 2H, J= 5.8 Hz, ArH), CHNH₂not seen.

DL-ritreo-2-Amino-3-hydroxy-3 -(pyridin-4-yl)-1 -(piperazin-1 -yl)propan-1 -one trihydrochloride Compound 37.

Compound 37 was prepared following method E with Zra»5-(4,5-dihydro5-(pyridin-4-yl)oxazol-4-yl)(4-tez7-butyloxycarbonyl-piperazin-l-yl)methanone SLA 07124B (0.31 g, 0.86 mmol), hydrochloric acid 37 % (1.0 mL) and

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2018203022 01 May 2018 methanol (10 mL). After 3 h at 50 °C and work-up bL-7zreo-2-amino-3hydroxy-3-(pyridin-4-yl)-1 -(piperazin-1 -yl)propan~ 1 -one trihydrochloride Compound 37 was obtained as a yellow solid (0.303 g, 71 %).

oh o (+/-)

NH₂ k^-NH

3.HCI

Compound 37

MW: 359.8; Yield: 71 %; Yellow Solid; Mp (°C): 201.4.

Rf. 0.20 (CH₂Cl₂:MeOH = 90:10), free base.

Ή-NMR (CD₃OD, δ): 3.31-3.48 (m, 4H, 2xCH₂), 3.63-3.90 (m, 2H, CH₂N), 4.00-4.35 (m, 2H, CH₂N), 5.15 (d, 1H, J= 4.5 Hz, CHNH₂), 5.58 (d, 1H, J= 4.5 Hz, CHO), 8.38 (d, 2H, J= 6.4 Hz, ArH), 9.04 (d, 2H, J= 6.5 Hz, ArH).

DL-/Areo-2-amino-3-hydroxy-3-(pyridin-4-yl)-l-thiomorpholinopropan-l-one dihydrochloride Compound 38.

Compound 38 was prepared following method E with /raw.s^,-(4,5-dihydro5-(pyridin-4-yl)oxazol-4-yl)(thiomorpholino)methanone SLA 07132 (0.926 g, 3.36 mmol), hydrochloric acid 37 % (1.1 mL) and methanol (10 mL). After 3 h at 50 °C and work-up DL-//weo-2-amino-3-hydroxy-3-(pyridin-4-yl)-lthiomorpholinopropan-l-one dihydrochloride Compound 38 was obtained as a pale yellow solid (1.1 g, 99 % yield).

OH O

Compound 38

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MW: 340.35; Yield: 99 %; Pale Yellow Solid; Mp (°C): 200.6.

Ή-NMR (CD₃OD, δ): 2.42-2.52 (m, 1H, CH₂), 2.53-2.70 (m, 1H, CH₂), 2.70-

2.90 (m, 2H, CH₂), 3.45-3.71 (m, 2H, CH₂N), 3.87-4.00 (m, 1H, CH₂N), 4.18-

4.28 (m, 1H, CH₂N), 5.44 (d, 1H, J= 5.1 Hz, CHO), 8.34 (d, 2H, J = 5.9 Hz, ArH), 9.03 (d, 2H, 5.6 Hz, ArH), -CHNH₂ not seen (under H₂O).

¹³C-NMR (CD₃OD, δ): 27.9, 28.7, 46.6, 50.0, 56.0, 71.3, 126.9 (2xC), 143.2 (2xC), 161.3, 165.7.

DL-/Areo-2-Amino-3-hydroxy-3-(pyridin-4-yl)-1 -(2J/-pyrrol-1 (5J7)-yl)propan-

1-one dihydrochloride Compound 39.

Compound 39 was prepared following method E with trans-(fi,5-dihydro5-(pyridin-4-yl)oxazol-4-yl)(27Z-pyrrol-l(5Z/)-yl)methanone SLA 07180 (0.276 g, 1.14 mmol), hydrochloric acid 37 % (1.0 mL) and methanol (10 mL). After 2.5 h at RT and work-up DL-//zreo-2-amino-3-hydroxy-3-(pyridin-4-yl)-l-(2#pyrrol-l(5/Z)-yl)propan-l-one dihydrochloride Compound 39 was obtained (343 mg, 99 % yield) as a white solid.

oh o nh₂ (+/-) 2. HCI

Compound 39

MW: 306.27; Yield: 99 %; White Solid; Mp (°C): 186.3.

'H-NMR (CD₃OD, δ): 3.91-4.02 (m, 1H, CH-NH₂), 4.09-4.21(m, 1H, CH₂), 4.27-4.41 (m, 1H, CH₂), 4.44-4.59 (m, 1H, CH₂), 4.55 (d, 1H, J = 5.7 Hz, CH₂N), 5.46 (d, 1H, J = 5.7 Hz, CHO), 5.80-5.90 (m, 2H, CH=CH), 8.24 (d, 1H, J= 6.3 Hz, ArH), 8.93 (d, 1H, J= 5.7 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 54.6, 54.7, 57.6, 71.0, 125.9, 126.4, 126.8 (2xC), 143.1 (2xC), 161.6, 165.5.

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MS-ESI m/z (% rel. Int.): 234.1 ([MH]⁺, 5), 137.1 (100).

DL-rAreo-2-Amino-7V,W-diethyl-3-hydroxy-3-(pyridin-4-yl)propanamide dihydrochloride Compound 40.

Compound 40 was prepared following method E with ^’izzz^-JVjTV-diethyl-

4,5-dihydro-5-(pyridin-4-yl)oxazole-4-carboxamide diethylamide SLA 07194A (254 mg, 1.03 mmol), hydrochloric acid 37 % (1.0 mL) and methanol (10 mL). After 2 h at RT and work-up DL-/Areo-2-amino-7V,W-diethyl-3-hydroxy-3(pyridin-4-yl)propanamide dihydrochloride Compound 40 was obtained (212 mg, 67 % yield) as a pale yellow solid.

OH O nh₂ (+/-) 2. HCI

Compound 40

MW: 310.30; Yield: 67 %; Pale Yellow Solid; Mp (°C): 159.6°C.

Ry. 0.10 (CH₂Cl₂:MeOH = 90:10), free base.

‘H-NMRCCDsOD, δ): 1.01-1.12 (m, 6H, 2xCH₃), 3.01-3.31 (m, 3H, CH₂),

3.40-3.52 (m, 1H, CH₂), 4.64 (d, 1H, J= 6.8 Hz, CHN), 5.31 (d, 1H, 7= 6.8 Hz, CHO), 8.22 (d, 1H, 7= 6.4 Hz, ArH), 8.94 (d, 1H, 7= 6.4 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 12.9, 14.4,42.1,43.4, 55.9, 72.1, 126.9 (2xC), 143.3 (2xC), 161.5, 166.1.

MS-ESI m/z (% rel. Int.): 238.1 ([MH]⁺, 5), 137.1 (100).

DL-rAreo-2-Amino-3 -(2-chloropyridin-4-yl)-3-hydroxy-1 -(pyrrolidin-1 yl)propan-l-one dihydrochloride Compound 41.

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Compound 41 was prepared following method E with /77^-(5-(2chloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1 -yl)methanone SLA 07174 (0.179 g, 0.64 mmol), hydrochloric acid 37 % (1.0 mL) and methanol (7 mL). After 2 h at RT and work-up DL-/Z?reo-2-amino-3-(2-chloropyridin-4-yl)-

3-hydroxy-l-(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 41 was obtained (212 mg, 67 (142 mg, 65 % yield) as a pale yellow solid.

OH O

(+/-) 2. HCI

Compound 41

MW: 342.65; Yield: 65 %; Pale Yellow Solid; Mp (°C): 184.3.

Ry. 0.15 (CH₂Cl₂:MeOH = 90:10), free base.

^-NMR (CD₃OD, δ): 1.50-1.90 (m, 4H, 2xCH₂), 2.50-2.61 (m, 1H, CH₂N),

3.25-3.38 (m, 1H, CH₂N), 3.40-3.53 (m, 2H, CH₂N), 4.26 (d, 1H, J= 7.7 Hz, CHN), 4.99 (d, 1H, 7.7 Hz, CHO), 7.45 (d, 1H, J = 4.5 Hz, ArH), 7.53 (s,

1H, ArH), 8.41 (d, 1H, 4.9 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 24.9, 26.7,47.5,48.0, 58.4, 71.9,122.0, 123.5, 151.1, 152.7,154.0, 165.7.

MS-ESI m/z (% rel. Int.): 270.1/272.1 ([MH]⁺, 40/13), 171.0/172.0 (100/32).

DL-/Areo-2-Amino-3-(3-bromopyridin-4-yl)-3-hydroxy-1 -(pyrrolidin-1 vl)propan-l-one dihydrochloride Compound 42.

Compound 42 was prepared following method E with trans-(5-(3bromopyridin-4-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1 -yl)methanone BAL 01028A (1.141 g, 3.52 mmol), hydrochloric acid 37 % (0.6 mL) and methanol (15 mL). After 3 h at 50 °C and work-up DL-/Areo-2-amino-3-(3-bromopyridinWO 2006/081273

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4-yl)-3-hydroxy-l-(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 42 was obtained as a white solid (667 mg, 49 % yield).

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Br OH 0

(+/-) 2. HCI

Compound 42

MW: 387.10; Yield: 49.0 %; White Solid; Mp (°C): 216.3. ’HNMR^DaOD, δ): 1.73-1.99 (m, 4H, 2xCH₂), 3.01-3.05 (m, 1H, CH₂N), 3.44-3.51(m, 2H, CH₂N), 3.60-3.73(m, 1H, CH₂N), 4.60 (d, 1H, J= 5.5 Hz, CH-N), 5.54 (d, 1H, J= 5.5 Hz, CH-O), 8.26 (d, 1H, J= 5.7 Hz, ArH)), 8.86 (d, 1H, J= 5.7 Hz, ArH), 9.10 (s, 1H, ArH).

¹³C-NMR (CD₃OD, δ): 24.8, 27.1, 56.4, 70.1, 122.7, 127.6, 145.1, 148.4, 156.8, 165.0,2xC not seen.

MS-ESI m/z (% rel. Int.): 314.1/316.1 ([MH]⁺, 35/35), 215.0/217 (50/50). HPLC: Method A, detection UV 254 nm, Compound 42 RT = 3.08 min, peak area 92.8 %.

DL-f77reo-2-Amino-3-(3-chloropyridin-4-yl)-3-hydroxy-1 -(pyrrolidin-1 vl)propan-l-one dihydrochloride Compound 43.

Compound 43 was prepared following method E with fiw.s-(5-(3chloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1 -yl)methanone BAL 01028B (0.925 g, 3.31 mmol), hydrochloric acid 37 % (0.6 rnL) and methanol (15 mL). After 2 h at 50 °C and work-up DL-/Areo-2-amino-3-(3-chloropyridin-

4-yl)-3-hydroxy-1-(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 43 was obtained as a white solid (599 mg, 53 % yield).

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Cl OH O (+/-) 2. HCI

Compound 43

MW: 342.65; Yield: 53 %; White Solid; Mp (°C): 214.0.

’H NMR (CD₃OD, δ): 1.75-2.02 (m, 4H, 2xCH₂), 3.13-3.25 (m, 1H, CH₂N), 3.39-3.62(m, 2H, CH₂N), 3.65-3.80 (m, 1H, CH₂N), 4.66 (d, 1H, J= 4.5 Hz, CH-N), 5.66 (d, 1H, J= 4.6 Hz, CH-O), 8.40 (d, 1H, J = 5.8 Hz, ArH), 8.93 (d, 1H, J= 5.8 Hz, ArH), 9.13 (s, 1H, ArH).

¹³C-NMR (CD₃OD, δ): 23.3,25.6, 46.6,46.8, 54.6, 66.5, 126.5, 132.4, 141.7,

142.9,155.6, 163.4.

MS-ESI m/z (% rel. Int.): 270/272 ([MH]⁺, 33/11), 171/173 (100/32). HPLC: Method A, detection UV 254 nm, Compound 43 RT = 2.80 min, peak area 97.2 %.

DL-/7zreo-3 -Hydroxy-1 -oxo-3 -(1 -oxy-pyridin-4-yl)-1 -(pyrrolidin-1 -yl)propan-2ylcarbamate BAL 01060.

To a solution of DL-/Areo-3-hydroxy-l-oxo-3-(pyridin-4-yl)-l(pyrrolidin-l-yl)propan-2-ylcarbamate (300 mg, 0.81 mmol, free base obtained from Compound 58 by K₂CO₃, CH₂C1₂ treatment) in dichloromethane (40 mL) was added MCPBA (350 mg, 2.03 mmol). The resulting mixture was stirred overnight at room temperature. The mixture was concentrated and the crude product was purified by column chromatography (EtOAc:MeOH = 70:30). DLiAreo-3-Hydroxy- l-oxo-3-( 1 -oxy-pyridin-4-yl)-1 -(pyrrolidin-1 -yl)propan-2ylcarbamate BAL 01060 was obtained as a white solid (292 mg, 94 % yield).

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MW: 385.41; Yield: 94 %; White Solid.

Ή NMR CDC1₃, δ): 1.71-2.00 (m, 4H, 2xCH₂), 3.35-3.53 (m, 3H, CH₂N), 3.54-

3.68 (m, 1H, CH₂N), 4.65 (dd, 1H, J= 9.6 Hz, J= 2.0 Hz, CH-N), 4.90-5.12 (m, 3H, CH₂O & OH), 5.20 (d, 1H, J= 1.9 Hz, CH-O), 5.83 (d, 1H, J= 9.6 Hz, NH), 7.20-7.40 (m, 7H, ArH), 8.08 (d, 2H, J= 7.1 Hz, ArH).

DL-/^reo-2-Amino-3-hydroxy-3-( 1 -oxy-pyridin-4-yl)-1 -pyrrolidin-1 -ylpropan-l-one hydrochloride Compound 44, [2-Hydroxy-2-( 1 -oxy-pyridin-4-yl)-1 -(pyrrolidine-1 -carbonyl)-ethyl]carbamic acid benzyl ester BAL 01060 (0.26 g, 0.67 mmol) was dissolved in a 6 N hydrochloric acid solution (10 mL). The solution was stirred for 0.75 h at 100 °C. The residue was concentrated, dissolved in MeOH:EtOAc = 50:50 and heated at reflux. After cooling, the mixture was evaporated, triturated in MeOH and filtered to obtain DL-/Areo-2-amino-3-hydroxy-3-(l-oxy-pyridin-4-yl)-lpyrrolidin-l-yl-propan-l-one hydrochloride Compound 44 (65 mg, 33 % yield) as a white solid.

oh 0

HCI (+/-)

Compound 44

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MW: 287.74; Yield: 33 %; White Solid; Mp (°C): 178.5.

Ή NMR (D₂O, δ): 1.55-1.93 (in, 4H, 2xCH₂), 2.65-3.80 (m, 1H, CH₂N), 3.223.56 (m, 3H, CH₂N), 4.43 (d, 1H, 7.6 Hz, CH-N), 5.19 (d, 1H, J= 7.6 Hz,

CH-O), 7.69 (d, 2H, 6.1 Hz, ArH), 8.39 (d, 2H, 6.9 Hz, ArH).

¹³C-NMR (D₂O, δ): 24.1, 25.8, 47.3, 48.0, 57.4, 70.5, 125.7 (2xC), 139.9 (2xC),

143.7,165.1.

MS-ESI m/z (% rel. Int.): 252.1 ([MH]⁺, 18), 120.0 (100).

HPLC: Method A, detection UV 254nm, Compound 44 RT = 0.8 min, peak area

99.9 %.

DL-7/zreo-2-(Dimethylamino)-3 -hydroxy-3 -(pyridin-4-yl)-1 -(pyrrolidin-1 yl)propan-l-one dihydrochloride Compound 45.

OL-threo-2-Amino-3 -hydroxy-3-(pyridin-4-yl)-1 -(pyrrolidin-1 yl)propan-l-one dihydrochloride Compound 22 (0.50 g, 1.62 mmol) and para ‘ formaldehyde (0.245 g, 8.11 mmol) were stirred in methanol (25 mL) for 10 min. Sodium cyanoborohydride (0.612 g, 9.73 mmol) was added. The solution was stirred 19 h at 50 °C and then concentrated. The residue was partitioned between dichloromethane and water. The aqueous layer was basified with IN sodium hydroxyde (pH = 10). The organic layer was combined with additional dichloromethane extracts, washed with aqueous sodium chloride and dried with MgSO4. The crude product was purified by column chromatography on silica (CH₂Cl₂:MeOH = 95:05). DL-^reo-2-(dimethylamino)-3-hydroxy-3-(pyridin4-yl)-l-(pyrrolidin-l-yl)propan-l-one SLA 07140 was obtained (187 mg, 44 %) as a yellow oil. To a stirred solution of DL-i/zreo-2-(dimethylamino)-3-hydroxy-

3-(pyridin-4-yl)-l-(pyrrolidin-l-yl)propan-l-one SLA 07140 (0.142 g, 0.54 mmol) in ethyl acetate (5 mL) was added dropwise via syringe 4 mL of a solution of HCI in Et₂O (0.3 M). The reaction mixture was stirred at 0 °C for 0.5

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h. The precipitate was filtered, washed with Et₂O and dried. OL-threo-2(Dimethylamino)-3-hydroxy-3 -(pyridin-4-yl)-1 -(pyrrolidin-1 -yppropan-1 -one dihydrochloride Compound 45 was obtained (0.077 g, 43 % yield) as a white solid.

OH o

II £ 7

NxV Vk ^L (+/-) 2 HCI

Compound 45

MW: 336.34; Yield: 43 %; White Solid; Mp (°C): 201.0.

Ή-NMR (CD₃OD, δ: 1.48-1.64 (m, 2H, CH₂), 1.65-1.83 (m, 2H, CH₂), 2.60-

2.72 (m, 1H, CH₂N), 3.15-3.33 (m, 1H, CH₂), 3.30-3.52 (m, 2H, CH₂), 4.60 (d, 1H, J= 8.4 Hz, CHNH₂), 5. 41 (d, 1H, 8.4 Hz, CHO), 8.10 (d, 2H, J= 6.6

Hz, ArH), 8.84 (d, 2H, J= 6.7 Hz, ArH).

DL-/Areo-2-Amino-1 -(pyridin-4-yl)-3-(pyrrolidin-1 -yPpropan-1 -ol Compound 4fr

To a stirred suspension of DL-i/zreo-2-amino-3-hydroxy-3-(pyridin-4-yl)-

1-(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 22 (0.86 g, 2.80 mmol) in tetrahydrofuran (108 mL) under nitrogen atmosphere was slowly added, in two portions, lithium aluminium hydride (0.64 g, 16.82 mmol) at 0 °C. The mixture reaction was stirred at RT for 20 h and quenched by a slow, dropwise addition of 2 N aqueous sodium hydroxyde (8.4 mL, 6 eq). The yellow precipitate was filtered. The organic layer was washed by water (80 mL) and the organic layer was removed and combined with additional ethyl acetate extracts (4 x 200 mL) and dried over MgSO₄, filtered and evaporated. The crude product was purified by column chromatography on silica (CH₂Cl₂:MeOH:NH3 = 94:05:01). After evaporation and drying DL-^reo-2-amino-l-(pyridin-4-yl)WO 2006/081273

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3-(pyrrolidin-l-yl)propan-l-ol Compound 46 was obtained (0. 075 g, 12 % yield) as a pale yellow solid.

Compound 46

MW: 221.30; Yield: 12 %; Pale Yellow Solid.

Ry. 0.35 (CH₂Cl₂:MeOH:NH₃ = 90:08:02).

Ή-NMR (CD₃OD, δ): 1.60-1.80 (m, 4H, 2xCH₂), 2.30-2.80 (m, 6H, 3xCH₂N), 3.14-3.19 (m, 1H, CHNH₂), 4.68 (d, 1H, J= 3.0 Hz, CHO), 7.30 (d, 2H, J= 6.0 Hz, ArH), 8.55 (d, 2H, J= 6.0 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 23.5 (2xC), 54.1, 54.7 (2xC), 60.1, 74.5, 121.4 (2xC), 149.5 (2xC), 152.1.

MS-ESI m/z (rel. int.): 222.1 ([MH]+, 100), 205.0 (80), 189.0 (45), 151.0 (70), 134.0 (42), 121.9 (100), 107.9 (40).

DL-/Areo-2-Amino-3-hydroxy-3-(2-methoxypyridin-3-yl)-l-(pyrrolidin-lyl)propan-l-one dihydrochloride Compound 48.

Trans-(4,5-Dihydro-5-(2-methoxypyridin-3-yl)oxazol-4-yl)(pyrrolidin-lyl)methanone BAL 01014 (0.465 g, 1.69 mmol) was dissolved in methanol (6 mL). The solution of hydrochloric acid (37 %, 0.3 mL) was added via a syringe at RT. The mixture was stirred for 3 h at RT. The residue was concentrated, dissolved in the minimum of MeOH, precipitated with EtOAc and filtered to obtain a white solid DL-/Areo-2-amino-3-hydroxy-3-(2-methoxypyridin-3-yl)-l(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 48 (103 mg, 18.0 % yield).

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Compound 48

MW: 338.23; Yield: 18.0 %; White Solid; Mp (°C): 171.5.

^!HNMR (CD₃OD, δ): 1.85-2.10 (m, 4H, CH₂), 3.30-3.82 (m, 4 H, CH₂N), 4.26 (s, 3H, OCH₃), 4.60 (d, 1H, J= 3.7 Hz, CH-N), 5.45 (d, 1H, J= 3.7 Hz, CH-O), 7.39 (dd, 1H, J= 5.6 Hz, J= 7.3, ArH), 8.32 (dd, 1H, 5.6 Hz, J= 7.3 Hz,

ArH).

¹³C-NMR (CD3OD, δ): 24.9, 27.1, 47.8,47.9, 56.4, 56.7, 66.0, 119.2, 125.6,

141.5,143.9, 160.6, 166.1.

MS-ESI m/z (% rel. Int.): 266.2 ([MH]⁺, 30), 248.2.0 (100). HPLC: Method A, detection UV 254nm, Compound 48 RT = 3.31 min, peak area 97.9 %.

3-(DL-/Zzreo-2-Amino- l-hydroxy-3-oxo-3-pyrrolidin-1 -yl-propyl)- lH-pyridin-

2-one hydrochloride Compound 49.

Erans-(4,5-Dihydro-5-(2-methoxypyridin-3-yl)oxazol-4-yl)(pyrrolidin-lyl)methanone BAL 01014 (0.684 g, 2.487 mmol) was dissolved in methanol (10 mL). A solution of hydrochloric acid (37 %, 0.6 mL) was added via syringe at RT. The mixture was stirred for 22 h at reflux. The residue was concentrated, triturated with EtOAc and filtered to obtain a yellow pale solid 3-(DL-//zreo-2amino-1 -hydroxy-3-oxo-3-pyrrolidin-1 -yl-propyl)- 177-pyridin-2-one hydrochloride Compound 49 (136 mg, 19.0 % yield).

OH 0

HN

HCI

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Compound 49

MW: 287.74; Yield: 19.0 %; Yellow Pale Solid; Mp (°C): 180.

^!H NMR (CD₃OD, δ): 1.82-2.09 (m, 4H, 2xCH₂), 3.35-3.80 (m, 4 H, 2xCH₂N), 4.63 (s, 1H, CH-N), 5.17 (s, 1H, CH-O), 6.56 (t, 1H, ArH)), 7.5 (d, 1H, J= 6.1 Hz, ArH), 7.86 (d, 1H, 6.5 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 24.2,26.0, 46.6,46.6, 75.8, 79.7,127.3, 127.5,127.9,

129.4, 130.0, 132.3, 133.2, 148.1,148.4, 155.3, 166.2.

MS-ESI m/z (% rel. Int.): 252.1 ([MH]⁺, 18), 163.0 (100). HPLC: Method A, detection UV 254nm, Compound 49 RT = 1.13 min, peak area 84.0 %.

Preparation of Compound 51, Compound 52, Compound 53, Compound 54 Compound 55, Compound 56 and Compound 57.

General procedures:

Method F:

To a suspension of DL-i/zreo-2-amino-3-hydroxy-3-(pyridin-4-yl)-l(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 22 (0.150 g, 0.44 mmol) in CH₂C1₂ (4 mL) was added TEA (0.185 mL, 1.32 mmol) and the reaction mixture was stirred for 10 min and cooled in an ice bath with continuous stirring. The acyl chloride (0.484 mmol) was dissolved in CH₂C1₂ (1 mL) and added dropwise to the reaction mixture. The reaction mixture was allowed to reach room temperature, stirred for 16 h and partitioned with H₂O (3 x 4 mL), washed with brine (3x4 mL), NaOH (0.5 M, 3 x 4 mL) and the organic layer was evaporated, adsorbed on silica gel (0.3 g) with EtOAc. The desired product was isolated by column chromatography using a gradient 0 to 8

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2018203022 01 May 2018 % [v/v] MeOH in EtOAc. The solid obtained was dissolved in ethanol (1 mL) and a solution of HCI (0.8 Μ, 1 mL) in EtOH was added. Evaporation of the volatiles led to the corresponding hydrochloride salt.

N-(OL-threo-1 -Hydroxy-3 -oxo-1 -(pyridin-4-yl)-3-(pyrrolidin-1 -yl)propan-2vDhexanamide hydrochloride Compound 51.

The compound was prepared according to method F with hexanoyl chloride (59 mg, 0.484 mmol) and DL-i/zreo-2-amino-3-hydroxy-3-(pyridin-4yl)-l-(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 22. ;V-(DLthreo-1 -Hydroxy-3 -oxo-1 -(pyridin-4-yl)-3 -(pyrrolidin-1 -yl)propan-2yl)hexanamide hydrochloride Compound 51 was obtained as an off white solid. (56 mg, 34 % yield).

OH o

HCI (+/-)

Compound 51

MW: 369.89; Yield: 34 %; Off White Solid; Mp (°C): 182.0.

^-NMR (CD₃OD, δ): 0.84 (t, 3H, J= 6.7, CH₃), 1.10-1.32 (m, 4H, CH₂), 1.351.50 (m, 2H, CH₂), 1.80-2.00 (m, 4H, CH₂), 2.05-2.30 (m, 2H, CH₂), 3.35-3.45 (m, 2H, CH₂), 3.50-3.65 (m, 2H, CH₂), 5.09 (d, 1H, J= 3.7 Hz, N-CH), 5.38 (d, 1H, J= 3.7 Hz, O-CH), 8.14 (d, 2H, J= 6.3 Hz, ArH), 8.80 (d, 2H, 6.3 Hz,

ArH).

MS-ESI m/z (% rel. int.): 334.2 ([MH]⁺, 10).

HPLC: Method A, detection UV 214 nm, Compound 51 RT = 3.90 min, peak area 99.0 %.

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100

N-(OL-threo-1 -Hydroxy-3 -oxo-1 -(pyridin-4-yP-3-(pyrrolidin-1 -yl)propan-2vDheptanamide hydrochloride Compound 52.

The compound was prepared according to method F with heptanoyl chloride (72 mg, 0.484 mmol) and DL-Z/?/'eo-2-amino-3-hydroxy-3-(pyridin-4yl)-l-(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 22. 7V-(DLthreo-1 -Hydroxy-3-oxo-1 -(pyridin-4-yl)-3-(pyrrolidin-1 -yl)propan-2ypheptanamide hydrochloride Compound 52 was obtained as an off white solid. (192 mg, 66 % yield).

oh o

HCI

Compound 52

MW: 383.91; Yield: 66 %; Off White Solid; Mp (°C): 187.1.

Ή-NMR (CD₃OD, δ): 0.88 (t, 3H, /=3.7 Hz, CH₃), 1.15-1.37 (m, 6H, CH₂),

l. 37 (m, 2H, CH₂), 1.85-2.02 (m, 4H, CH₂), 1.18-2.27 (m, 2H, CH₂), 3.37-3.50 ( m, 2H, N-CH₂), 3.55-3.70 (m, 2H, NCH₂), 5.14 (d, 1H, N-CH), 5.42 (d, 1H, OCH), 8.19 (d, 2H, /= 6.3 Hz, ArH), 8.83 (d, 2H, J = 6.3 Hz, ArH).

¹³C-NMR(CD₃OD, δ): 14.4,23.6, 25.0,26.7,27.0, 29.9, 32.6, 36.4, 47.5, ,56.7,

72.6, 126.6, 142.0, 164.5, 169.2, 175.9.

MS-ESI m/z (% rel. Int.): 348.2 ([MH]⁺, 10).

HPLC: Method A, detection UV 254 nm, Compound 52 RT = 4.10 min, peak area 99.0 %.

N-(OL-threo-1 -Hydroxy-3-oxo-1 -(pyridin-4-yl)-3-(pyrrolidin-1 -yPpropan-2yDoctanamide hydrochloride Compound 53.

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101

The compound was prepared according to method F with octanoyl chloride (78 mg, 0.484 mmol) and DL-//zreo-2-amino-3-hydroxy-3-(pyridin-4yl)-l-(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 22. #-(DLthreo-1 -Hydroxy-3-oxo-1 -(pyridin-4-yl)-3 -(pyrrolidin-1 -yl)propan-2yl)octanamide hydrochloride Compound 53 was obtained as an off white solid. (131 mg, 75 % yield).

Compound 53

MW: 397.94; Yield: 75 %; Off White Solid; Mp (°C): 185.9.

’H-NMR (CD₃OD, δ): 0.91 (t, 3H, J = 6.4 Hz, CH₃), 1.12-1.37 (m, 8H, CH₂),

1.40-1.52 (m, 2H, CH₂), 1.81 (m, 4H, CH₂), 2.12-2.25 (m, 2H, CH₂), 3.40-3.52 (m, 2H, N-CH₂), 3.55-3.65 (m, 2H, N-CH₂), 5.14 (d, 1H, 7= 3.7 Hz, N-CH),

5.43 (d, 1H, J= 3.7 Hz, OCH), 8.19 (d, 2H, 7= 6.3 Hz, ArH), 8.84 (d, 2H, 7=

6.3 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 14.4,23.7, 24.9,25.0, 25.5, 26.8,27.0, 30.1, 30.2, 32.8, 34.3, 36.5,47.5, 56.8, 71.6, 72.6, 126.7,126.9, 142.1,143.7, 164.5, 169.2, 176.0.

MS-ESI m/z (% rel. Int.): 362.2 ([MH]⁺, 10).

HPLC: Method A, detection UV 254 nm, Compound 53 RT = 4.37 min, peak area 99.9 %.

N-(OL-threo-1 -Hydroxy-3 -oxo-1 -(pyridin-4-yl)-3-(pyrrolidin-1 -yl)propan-2vDpalmitamide hydrochloride Compound 54.

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102

The compound was prepared according to method F with palmitoyl chloride (133 mg, 0.484 mmol) and DL-/Areo-2-amino-3-hydroxy-3-(pyridin-4yl)-l-(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 22.7V-(DLthreo-1 -Hydroxy-3-oxo-1 -(pyridin-4-yl)-3 -(pyrrolidin-1 -yl)propan-2yl)palmitamide hydrochloride Compound 54 was obtained as an off white solid. (105 mg, 47 % yield).

Compound 54

MW: 510.15; Yield: 47 %; White Solid; Mp (°C): 185.9.

Ή-NMR (CD₃OD, δ): 0.92 (t, 3H, CH₃), 1.18 -1.42 (m, 24H, CH₂), 1.42-1.58 (m, 2H, CH₂), 1.85 (m, 4H, CH₂), 2.15 (m, 2H, CH₂), 3.41-3.50 (m, 2H, CH₂), 3.50-3.68 (m, 2H, CH₂), 5.14 (d, 1H, J= 3.5 Hz, N-CH), 5.42 (d, 1H, J= 3.5 Hz, O-CH), 8.18 (d, 2H, J= 6.0 Hz, ArH), 8.82 (d, 2H, J= 5.7 Hz, ArH). ¹³C-NMR (CD₃OD, δ): 14.4, 23.7, 25.0,26.8,27.0, 30.3, 30.4, 30.5, 30.6, 30.7,

30.8, 33.1, 36.4, 47.5, 56.8, 72.6, 126.6, 142.1, 164.5, 169.2, 175.9.

MS-ESI m/z (% rel. Int.): 474.2 ([MH]⁺, 40).

HPLC: Method A, detection UV 254 nm, Compound 54 RT = 6.36 min, peak area 97.0%.

N-(OL-threo-1 -Hydroxy-3-oxo-1 -(pyridin-4-yl)-3-(pyrrolidin-1 -yl)propan-2yDbenzamide hydrochloride Compound 55.

The compound was prepared according to method F with benzoyl chloride (141 mg, 0.484 mmol) and DL-/7zreo-2-amino-3-hydroxy-3-(pyridin-4yl)-l-(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 22. A-(DLWO 2006/081273

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103 threo-1 -Hydroxy-3-oxo-1 -(pyridin-4-yl)-3 -(pyrrolidin-1 -yl)propan-2yl)benzamide hydrochloride Compound 55 was obtained as an off white solid. (67 mg, 34 % yield).

oh o h-ci

Compound 55

MW: 375.85; Yield: 34 %; Off White Solid; Mp (°C): 212.

Ή-NMR (CD₃OD, δ): 1.69-1.91 (m, 4H, CH₂), 3.25-3.40 (m, 2H, N-CH₂),

3.40-3.58 (m, 2H, N-CH₂), 5.22 (d, 1H, 3.7 Hz, N-CH), 5.43 (d, 1H, J= 3.5

Hz, O-CH), 7.32 (t, 2H, J= 7.8 Hz, ArH), 7.40 (t, 1H, J= 6.9 Hz, ArH), 7.63 (d, 2H, J = 7.1 Hz, ArH), 8.08 (d, 2H, J = 6.6 Hz, ArH), 8.66 (d, 2H, J - 6.1 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 25.0, 27.1, 47.6, 57.6, 72.7, 126.6, 128.4, 129.7,133.3,

134.4, 142.1, 164.5, 169.0, 169.7.

MS-ESI m/z (% rel. Int.): 340.2 ([MH]⁺, 5).

HPLC: Method A, detection UV 254 nm, Compound 55 RT = 3.66 min, peak area 99.0 %.

N-(DL-threo-1 -Hydroxy-3-oxo-1 -(pyridin-4-yl)-3-(pyrrolidin-1 -yl)propan-2yl)-4-methoxy-benzamide hydrochloride Compound 56.

The compound was prepared according to method F with 4-methoxybenzoyl chloride (82 mg, 0.484 mmol) and DL-/Areo-2-amino-3-hydroxy-3-(pyridin-4yl)-l-(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 22.7V-(DLthreo-1 -Hydroxy-3-oxo-1 -(pyridin-4-yl)-3 -(pyrrolidin-1 -yl)propan-2-yl)-4WO 2006/081273

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104 methoxy-benzamide hydrochloride Compound 56 was obtained as an off white solid. (105 mg, 58 % yield).

oh o

O’compound 56

MW: 405.87; Yield: 58 %; Off White Solid; Mp(°C): 205.3 (dec).

’H-NMR (CD₃OD, δ): 1.82-2.08 (m, 4H, CH₂), 3.45-3.55 (m, 2H, CH₂-N), 3.60-3.70 (m, 2H, NCH₂), 3.86 (s, 3H, O-CH₃), 5.35 (d, 1H, J = 3.7 Hz, N-CH), 5.56 (d, 1H, J= 3.6 Hz, O-CH), 6.99 (dd, 2H, J= 6.9 Hz, J= 1.9 Hz), 7.76 (dd, 2H, J= 6.9 Hz, J = 1.9 Hz, ArH), 8.21 (d, 2H, J =6.6 Hz, ArH), 8.79 (d, 2H, J = 6.6 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 25.0,27.1,47.6, 56.0, 57.5, 72.7,114.9,115.2, 126.3,

126.6,130.4, 133.7, 142.1, 164.4, 164.5, 169.1.

MS-ESI m/z (% rel. Int.): 370.2 ([MH]⁺, 10).

HPLC: Method A, detection UV 254 nm, Compound 56 RT = 3.76 min, peak area 99 %.

3,4-Dichloro-A-(DL-/Zzre<?-1 -Hydroxy-3-oxo- l-(pyridin-4-yl)-3-(pyrrolidin-1 yl)propan-2-yl)benzamide Compound 57.

The compound was prepared according to method F with 3,4dichlorobenzoyl chloride (101 mg, 0.484 mmol) and DL-7zreo-2-amino-3hydroxy-3-(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)propan-1 -one dihydrochloride Compound 22. 3,4-Dichloro-W-(DL-/Areo-l-hydroxy-3-oxo-l-(pyridin-4-yl)-3WO 2006/081273

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105 (pyrrolidin-l-yl)propan-2-yl)benzamide Compound 57 was obtained as an off white solid. (92 mg, 55 % yield).

MW: 444.74; Yield: 55 %; Off White Solid; Mp (°C): 319.5 (dec).

^-NMR (CD₃OD, δ): 1.82-2.05 (m, 4H, CH₂), 3.40-3.70 (m, 4H, N-CH₂), 5.33 (d, 1H, J= 3.9 Hz, N-CH), 5.55 (d, 1H, J = 4.0 Hz, O-CH), 7.61-7.75 (m, 2H, ArH), 7.96 (d, 1H, 7= 1.5 Hz, ArH), 8.22 (d, 2H, 7= 6.4 Hz, ArH), 8.81 (d, 2H, 7= 6.0 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 25.0, 27.0, 57.8, 72.6, 126.6, 128.3, 130.7, 131.9, 133.8,

134.7, 137.2, 142.2, 164.3, 167.2, 168.8.

MS-ESI m/z (% rel. Int.): 408.0, ([MH]⁺, 10)

HPLC: Method A, detection UV 254 nm, Compound 57 RT = 4.28 min, peak area 99.9 %.

Preparation of Compound 58, Compound 59, Compound 60, Compound 61, Compound 62, Compound 63, Compound 64, Compound 65, Compound 66, Compound 67, Compound 68, Compound 69.

General procedures:

Method G (in CH₂C1₂):

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To a stirred solution ofDL-rizreo-2-amino-3-hydroxy-3-(pyridin-4-yl)-l(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 22 (0.15 g, 0.49 mmol) in 10 mL of CH₂C1₂ at +4 °C were added triethylamine (200 μΐ, 1.45 mmol) and very slowly acid chloride in 3 mL of CH₂C1₂. The mixture was stirred overnight at RT under nitrogen and then partitioned between CH2CI2 and 1 N aqueous sodium carbonate. The organic layer was evaporated and the obtained residue purified by column chromatography on silica (EtOAc:MeOH = 95:5). The hydrochloride salt was obtained in MeOH at 0 °C with 0.3 M HCI in diethylether to give after evaporation of solvents and drying the acylated compound.

Method H (in MeOH):

To a stirred solution of DL-/Areo-2-amino-3-hydroxy-3-(pyridin-4-yl)-l(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 22 (0.20 g, 0.65 mmol) in 3 mL of MeOH were added triethylamine (180 μΐ, 1.30 mmol) and aldehyde (or ketone). The mixture was stirred overnight at RT under nitrogen and then was added AcOH (200 gL, 3.2 mmol) and NaBH₃CN. After 5 h at 20 °C, MeOH was evaporated and the residue was partitioned between CH₂C1₂ and 1 N aqueous sodium carbonate. The organic layer was evaporated and the obtained residue was purified by column chromatography on silica (EtOAc:MeOH or CH₂Cl₂:MeOH). The hydrochloride salt was obtained in MeOH at 0 °C with 0.3 M HCI in diethylether to give after evaporation of solvents and drying the alkylated compound.

Benzyl DL-/Are<?-3 -hydroxy-1 -oxo-3-(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)propan-

2-ylcarbamate hydrochloride Compound 58.

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-oxo-3-(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)propan-2-ylcarbamate hydrochloride

Compound 58 was obtained as a white solid (90 mg, 46 % yield).

OH^N^

Compound 58

MW: 405.9; Yield: 46.0 %; White Solid; Mp (°C): 185.3.

Ry·: 0.38 (MeOH:EtOAc = 10:90) free base.

Ή-NMR (CD₃OD, δ): 1.87-2.03 (m, 4H, 2xCH2), 3.40-3.48 (m, 2H, CH₂N), 3.56-3.62 (m, 2H, CH₂N), 4.85-5.04 (m, 3H, CH₂O, CHO), 5.39 (d, 1H, J= 2.8 Hz, NH), 7.26-7.36 (m, 5H, ArH), 8.12 (d, 2H, J= 6.0 Hz, ArH), 8.69 (d, 2H, J = 6.0 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 25.0, 27.0, 47.5,48.0, 58.8, 67.9, 72.7, 126.6 (2xC),

129.1, 129.2, 129.5, 138.1, 141.9 (2xC), 158.1, 164.4, 169.2.

MS-ESI m/z (% rel. Int.): 370.1 ([MH]⁺, 15), 219.0 (100).

HPLC: Method A, detection UV 254 nm, Compound 58 RT = 4.10 min, peak area 99.8 %.

7V-(DL-/Areo-3 -hydroxy-1 -oxo-3-(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)propan-2yl)decanamide hydrochloride Compound 59.

The compound was prepared according to method G with decanoyl chloride (111 pL, 0.53 mmol). After work-up W(DL-^reo-3-hydroxy-l-oxo-3(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)propan-2-yl)decanamide hydrochloride Compound 59 was obtained as a white solid (115 mg, 55 % yield).

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OH N

Compound 59

MW: 425.99; Yield: 55 %; White Solid; Mp (°C): 184.8.

R_z: 0.22 (MeOH:EtOAc = 5:95) free base.

‘H-NMR (CD₃OD, δ): 0.90 (t, 3H, J= 7.0 Hz, CH₃), 1.26-1.34 (m, 12H, 6xCH₂), 1.42-1.50 (m, 2H, CH₂), 1.86-1.98 (m, 4H, 2xCH₂), 2.13-2.20 (m, 2H, CH₂CO), 3.41-3.46 (m, 2H, CH₂N), 3.52-3.61 (m, 2H, CH₂N), 5.12 (d, 1H,

3.8 Hz, CH), 5.40 (d, 1H, J= 3.7 Hz, CH), 8.16 (d, 2H, J= 6.5 Hz, ArH), 8.97 (d, 2H, J= 6.7 Hz, ArH).

¹³C-NMR(CD₃OD, δ): 14.4,23.7, 25.0,26.8,27.0, 30.3, 30.4, 30.6, 33.0, 36.5,

47.5, 56.8, 72.6, 126.6 (2xC), 142.1 (2xC), 164.4, 169.2, 175.9.

MS-ESI m/z (% rel. Int.): 390.1 ([MH]⁺, 20), 219.1 (100).

HPLC: Method A, detection UV 254 nm, Compound 59 RT = 4.9 min, peak area 99.5 %.

DL-/Arg<9-2-(Benzylamino)-3-hydroxy-3-(pyridin-4-yl)-1 -(pyrrolidin-1 yDpropan-l-one dihydrochloride Compound 60.

The compound was prepared according to method H with benzaldehyde (78 mg, 0.72 mmol). After column chromatography (EtOAc:MeOH = 95:5) and HCI treatment DL-rf/reo-2-(benzylamino)-3-hydroxy-3-(pyridin-4-yl)-1 (pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 60 was obtained as a white solid (114 mg, 46 % yield).

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Compound 60

MW: 398.33; Yield: 46 %; White Solid; Mp (°C): 131.5.

Ry·: 0.60 (MeOH:EtOAc = 10:90) free base.

Ή-NMR (CD₃OD, δ): 1.35-1.72 (m, 4H, 2xCH₂), 2.10-2.18 (m, 1H, CH₂N), 2.78-2.86 (m, 1H, CH₂N), 3.18-3.24 (m, 2H, CH₂N), 4.22 (d, 1H, J= 8.5 Hz, CH), 4.26-4.36 (m, 2H, CH₂N), 5.18 (d, 1H, 8.5 Hz, CH), 7.43-7.51 (m, 5H,

BzH), 7.86 (d, 2H, J= 6.6 Hz, ArH), 8.69 (d, 2H, J= 6.6 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 24.7, 26.3, 47.3,47.9, 51.4, 63.9, 72.4, 126.2, 130.3, 131.0, 131.4, 131.5, 148.9, 156.1, 163.9.

MS-ESI m/z (% rel. Int.): 326.1 ([MH]⁺, 100), 227.0 (80).

HPLC: Method A, detection UV 254 nm, Compound 60 RT = 4.30 min, peak area 98.2 %.

DL-/Z?reo-3-Hydroxy-2-(methylamino)-3-(pyridin-4-yl)-1 -(pyrrolidin-1 yl)propan-l-one dihydrochloride Compound 61.

The compound was prepared according to method H with paraformaldehyde (21 mg, 0.65 mmol). After colummn chromatography (EtOAc:MeOH = 7:3) and HCI treatment DL-/Areo-3-hydroxy-2(methylamino)-3 -(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)propan-1 -one dihydrochloride Compound 61 was obtained as a pale yellow solid (28 mg, 13 % yield).

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Compound 61

MW: 322.23; Yield: 13 %; Pale Yellow Solid.

Ry: 0.20 (MeOH:EtOAc = 30:70) free base.

^-NMR (CD₃OD, δ): 1.60-1.80 (m, 4H, 2xCH₂), 2.61 (s, 1H, CH₃), 2.68-2.76 (m, 1H, CH₂N), 3.24-3.57 (m, 3H, CH₂N), 4.53 (d, 1H, J= 6.8 Hz, CH), 5.26 (d, 1H, J= 7.0 Hz, CH), 8.11 (d, 2H, J= 5.8 Hz, ArH), 8.85 (d, 2H, J= 5.6 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 24.8, 26.7, 32.8,47.7,48.3, 65.4, 71.7, 126.7 (2xC), 143.4 (2xC), 161.1, 163.9.

MS-ESI m/z (% rel. Int.): 251.1 ([MH]⁺, 10), 151.0 (100).

HPLC: Method A, detection UV 254 nm, Compound 61 RT = 0.70 min, peak area 97.5 %.

DL-fAreo-3-Hydroxy-2-(pentylamino)-3-(pyridin-4-yl)-1 -(pyrrolidin-1 yDpropan-1-one dihydrochloride Compound 62,

The compound was prepared according to method H with valeraldehyde (60 mg, 0.68 mmol). After column chromatography (EtOAc:MeOH = 95:5) and HCI treatment OL-threo-3 -hydroxy-2-(pentylamino)-3-(pyridin-4-yl)-1 (pyrrolidin-1-yl)propan-1-one dihydrochloride Compound 62 was obtained as a white solid (107 mg, 44 % yield).

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MW: 378.34; Yield: 44 %; White Solid; Mp (°C): 86.6.

Ry: 0.30 (MeOH:EtOAc = 20:80) free base.

Ή-NMR (CD₃OD, δ): 0.95 (t, 3H, 6.4 Hz, CH₃), 1.32-1.40 (m, 6H, 3xCH2),

1.70-1.87 (m, 4H, 2xCH₂), 2.70-2.75 (m, 1H, CH₂N), 2.90-3.00 (m, 1H, CH₂N), 3.10-3.39 (m, 3H, CH₂N), 3.46-3.60 (m, 1H, CH₂N), 4.61 (d, 1H, J= 7.5 Hz, CH), 5.39 (d, 1H, 7.5 Hz, CH), 8.22 (d, 2H, 6.2 Hz, ArH), 8.97 (d, 2H, J = 6.2 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 9.3,14.1,23.2, 24.8,26.7, 26.8,29.7,47.6,47.9, 64.5,

72.1, 126.8 (2xC), 143.3 (2xC), 161.1, 164.1.

MS-ESI m/z (% rel. Int.): 306.3 ([MH]⁺, 15), 207.1 (100).

HPLC: Method A, detection UV 254 nm, Compound 62 RT = 3.60 min, peak area 98.5 %.

DL-zAreo-3 -Hydroxy-2-(hexylamino)-3-(pyridin-4-yl)-1 -(pyrrolidin-1 vl)propan-l-one dihydrochloride Compound 63.

The compound was prepared according to method H with hexanal (71 mg, 0.68 mmol). After column chromatography (EtOAc:MeOH = 95:5) and HCI treatment DL-/Areo-3-hydroxy-2-(hexylamino)-3-(pyridin-4-yl)-1 (pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 63 was obtained as a beige solid (112 mg, 45 % yield).

(+/-)

Compound 63

MW: 392.36; Yield: 45 %; Beige Solid; Mp (°C): 108.2.

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Ry·: 0.35 (MeOH:EtOAc = 20:80) free base.

Ή-NMR (CD₃OD, δ): 0.94 (t, 3H, J= 7.2 Hz, CH₃), 1.30-1.42 (m, 6H, 3xCH₂), 1.66-1.90 (m, 6H, 3xCH₂), 2.68-2.74 (m, 1H, CH₂N), 2.90-2.99 (m, 1H, CH₂N), 3.09-3.16 (m, 1H, CH₂N), 3.32-3.39 (m, 1H, CH₂N), 3.47-3.60 (m, 2H, CH₂N), 4.60 (d, 1H, J= 7.7 Hz, CH), 5.38 (d, 1H, J= 7.7 Hz, CH), 8.24 (s, 2H, ArH), 8.97 (s, 2H, ArH).

¹³C-NMR (CD3OD, δ): 14.3, 23.4, 24.8,26.7,27.1, 27.2, 32.4,47.6, 64.4, 72.1,

126.9 (2xC), 143.2 (2xC), 161.3, 164.1.

MS-ESI m/z (% rel. Int.): 320.1 ([MH]⁺, 30), 221.1 (100).

HPLC: Method A, detection UV 254 nm, Compound 63 RT = 3.80 min, peak area 97.8 %.

DL-/Z?reo-3-Hydroxy-2-(heptylamino)-3-(pyridin-4-yP-1 -(pyrrolidin-1 yppropan-1-one dihydrochloride Compound 64.

The compound was prepared according to method H with heptaldehyde (82 mg, 0.68 mmol). After column chromatography with EtOAc:MeOH = 95:5 and HCI treatment DL-/Areo-3-hydroxy-2-(heptylamino)-3-(pyridin-4-yl)-l(pyrrolidin-1-yppropan-1-one dihydrochloride Compound 64 was obtained as a white solid (121 mg, 47 % yield).

Compound 64

MW: 406.39; Yield: 47 %; White Solid; Mp (°C): 242.4.

Ry·: 0.40 (MeOH:EtOAc = 20:80) free base.

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113 ’H-NMR (CD₃OD, δ): 0.92 (t, 3H, J= 7.0 Hz, CH₃), 1.32-1.40 (m, 8H, 4xCH₂), 1.65-1.90 (m, 6H, 3xCH₂), 2.71-2.76 (m, 1H, CH₂N), 2.90-2.99 (m, 1H, CH₂N), 3.09-3.38 (m, 1H, CH₂N), 3.47-3.62 (m, 2H, CH₂N), 4.61 (d, 1H, 7.5 Hz,

CH), 5.39 (d, 1H, 7.5 Hz, CH), 8.24 (d, 2H, J= 6.0 Hz, ArH), 8.97 (d, 2H, J = 5.9 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 14.4,23.6, 24.8,26.7,27.1, 27.5,29.9, 32.7,47.7, 64.4,

72.1, 126.9 (2xC), 143.2 (2xC), 161.4, 164.1.

MS-ESI m/z (% rel. Int.): 334.1 ([MH]⁺, 45), 235.1 (100).

HPLC: Method A, detection UV 254 nm, Compound 64 RT = 4.00 min, peak area 97.5 %.

DL-/Areo-2-(4-Methylbenzylamino)-3-hydroxy-3-(pyridin-4-yl)-l-(pyrrolidin- l-yl)propan-l-one Compound 65.

The compound was prepared according to method H with 4methylbenzaldehyde (86 mg, 0.70 mmol). After column chromatography (EtOAc:MeOH = 95:5) and HCI treatment DL-/Areo-2-(4-methylbenzylamino)-

3-hydroxy-3-(pyridin-4-yl)-l-(pyrrolidin-l-yl)propan-l-one Compound 65 was obtained as a white solid (137 mg, 51 % yield).

MW: 412.35; Yield: 51 %; White Solid; Mp (°C): 87.5.

Ry·: 0.20 (MeOH:EtOAc = 5:95) free base.

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Ή-NMR (CD₃OD, δ): 1.49-2.12 (m, 4H, 2xCH₂), 2.30-2.40 (m, 1H, CH₂N), 2.35. (s, 3H, CH₃), 2.75-2.95 (m, 1H, CH₂N), 3.18-3.25 (m, 2H, CH₂N), 4.12-

4.32 (m, 3H, CH₂N, CH), 5.30 (d, 1H, J= 7.9 Hz, CH), 7.24-7.39 (m, 4H, BzH), 8.11 (d, 2H, J= 6.7 Hz, ArH), 8.89 (d, 2H, J= 6.6 Hz, ArH). ¹³C-NMR (CD₃OD, δ): 21.2, 24.7, 26.3,26.4,47.4,47.9, 51.2, 63.4, 72.4,

126.1, 126.6 (2xC), 128.2, 130.9, 131.4, 131.5, 141.4, 143.5 (2xC), 148.9, 156.1; 160.5, 163.8.

MS-ESI m/z (% rel. Int.): 340.1 ([MH]⁺, 10), 104.9 (100).

HPLC: Method A, detection UV 254 nm, Compound 65 RT = 3.70 min, peak area 97.3 %.

DL-/Zzreo-2-(4-Chlorobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1 -(pyrrolidin-1 vl)propan-l-one Compound 66.

The compound was prepared according to method H with 4chlorobenzaldehyde (98 mg, 0.70 mmol). After column chromatography (EtOAc:MeOH = 95:5) and HCI treatment DL-/Zzreo-2-(4-chlorobenzylamino)-

3-hydroxy-3-(pyridin-4-yl)-l-(pyrrolidin- l-yl)propan-l-one Compound 66 was obtained as a white solid (126 mg, 45 % yield).

OH N

Cl

Compound 66

MW: 432.77; Yield: 45 %; White Solid; Mp (°C): 122.7.

Ry: 0.20 (MeOH: EtOAc = 5:95) free base.

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Ή-NMR (CD3OD, δ): 1.54-1.75 (m, 4H, 2xCH₂), 2.41-2.49 (m, 1H, CH₂N), 3.08-3.15 (m, 1H, CH₂N), 3.19-3.27 (m, 2H, CH₂N), 4.25-4.47 (m, 3H, CH₂N, CH), 5.34 (d, 1H, J= 7.9 Hz, CH), 7.46-7.55 (m, 4H, BzH), 8.15 (d, 2H, J= 6.0 Hz, ArH), 8.92 (d, 2H, J= 5.7 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 24.7, 26.5,47.5,47.8, 50.8, 63.8, 72.3, 126.7 (2xC),

130.3, 133.4, 137.1, 143.5 (2xC), 149.0, 160.6, 163.8.

MS-ESI m/z (% rel. Int.): 360.1/362.1 ([MH]⁺, 20), 124.9 (100).

HPLC: Method A, detection UV 254 nm, Compound 66 RT = 3.70 min, peak area 97.0 %.

DL-/AFeo-2-(4-Methoxybenzylamino)-3-hydroxy-3-(pyridin-4-yl)-l-(pyrrolidin- l-vl)propan-l-one one Compound 67.

The compound was prepared according to method H with 4methoxybenzaldehyde (95 mg, 0.70 mmol). After column chromatography (EtOAc:MeOH = 95:5) and HCI treatment OL-threo-2-(4methoxybenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)propan-1 one one Compound 67 was obtained as a white solid (123 mg, 44 % yield).

MW: 428.35; Yield: 44 %; White Solid; Mp (°C): 193.2.

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Ry·: 0.20 (MeOH:EtOAc = 5:95) free base.

*H-NMR (CD₃OD, δ): 1.51-1.72 (m, 4H, 2xCH₂), 2.38-2.41 (m, 1H, CH₂N), 2.94-3.01 (m, 1H, CH₂N), 3.18-3.28 (m, 2H, CH₂N), 3.81 (s, 3H, CH₃O), 4.18-

4.34 (m, 3H, CH₂N, CH), 5.31 (d, 1H, 7.3 Hz, CH), 6.97 (d, 2H, J= 8.5 Hz,

ArH), 7.42 (d, 2H, 8.5 Hz, ArH), 8.13 (d, 2H, J= 6.5 Hz, ArH), 8.91 (d, 2H,

J= 6.3 Hz, ArH).

¹³C-NMR(CD₃OD, δ): 24.7,26.5, 47.5,47.9, 51.1, 55.9, 63.2, 72.3, 115.5,

122.9, 126.8 (2xC), 133.1, 143.3 (2xC), 160.9, 162.4, 163.8.

MS-ESIm/z (%rel. Int.); 356.1 ([MH]⁺, 10), 120.9 (100).

HPLC: Method A, detection UV 254 nm, Compound 67 RT = 3.50 min, peak area 98.6 %.

OL-threo-2-(2,4-Dichlorobenzylamino)-3-hydroxy-3 -(pyridin-4-yl)-1 (pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 68.

The compound was prepared according to method H with 3,4dichlorobenzaldehyde (122 mg, 0.70 mmol). After column chromatography (EtOAc:MeOH = 95:5) and HCI treatment DL-/Z?reo-2-(3,4dichlorobenzylamino)-3 -hydroxy-3-(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)propan-1 one dihydrochloride Compound 68 was obtained as a white solid (153 mg, 50 % yield).

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MW: 467.22; Yield: 50 %; White Solid; Mp (°C): 190.3.

Ry: 0.20 (MeOH.'EtOAc = 5:95) free base.

’H-NMR (CD₃OD, δ): 1.58-1.76 (m, 4H, 2xCH₂), 2.48-2.55 (m, 1H, CH₂N),

2.85-3.00 (m, 1H, CH₂N), 3.18-3.26 (m, 2H, CH₂N), 4.23-4.41 (m, 2H, CH₂N), 4.54 (d, 1H, J= 7.7 Hz, CH), 5.34 (d, 1H, J= 7.2 Hz, CH), 7.48 (d, 1H, J= 8.3 Hz, ArH), 7.63 (dd, 1H, J= 8.2 Hz, 7=1.4 Hz, ArH), 7.75 (s, 1H, ArH), 8.16 (d, 2H, 7= 5.4 Hz, ArH), 8.92 (d, 2H, 7= 5.3 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 23.2,25.0,46.0,46.4,48.9, 62.5, 70.7, 125.3, 130.2,

130.6, 130.7, 132.3, 133.6,141.9, 159.2, 162.3.

MS-ESI m/z (% rel. Int.): 394.1/396.1 ([MH]⁺, 40), 110.0 (100).

HPLC: Method A, detection UV 254 nm, Compound 68 RT = 3.90 min, peak area 99.0 %.

DL-//zreo-2-(4-Methoxybenzylamino)-3-hydroxy-3-(pyridin-4-yl)-l-(pyrrolidin1-yDpropan-l-one Compound 69.

The compound was prepared according to method H with cyclohexanone (75 pL, 0.70 mmol). After column chromatography (EtOAc:MeOH = 95:5) and HCI treatmentDL-/Areo-2-(4-methoxybenzylamino)-3-hydroxy-3-(pyridin-4yl)-l-(pyrrolidin-l-yl)propan-l-one Compound 69 as a white solid (154 mg, 61 % yield).

Compound 69

MW: 390.35; Yield: 61 %; White Solid; Mp (°C): 144.1.

Ry: 0.25 (MeOH: EtOAc = 5:95) free base.

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Ή-NMR (CD₃OD, δ): 1.16-2.15 (m, 15H, 7xCH₂, CH), 2.65-2.72 (m, 1H, CH₂N), 3.07-3.15 (m, 1H, CH₂N), 3.43-3.65 (m, 2H, CH₂N), 4.61 (d, 1H, J=

7.8 Hz, CH-N), 5.35 (d, 1H, J= 7.8 Hz, CH-O), 8.21 (d, 2H, J = 6.3 Hz, ArH), 8.95 (d, 2H, J =6.1 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 23.2, 24.0, 24.2,24.4, 25.2, 28.2, 29.3, 46.1, 57.4, 60.5,

71.0, 125.4 (2xC), 141.8 (2xC), 159.5, 162.6.

MS-ESI m/z (% rel. Int.): 318.2 ([MH]⁺, 40; 219.1,100). HPLC: Method A, detection UV 254 nm, Compound 69 RT = 3.40 min, peak area 99.7 %.

Preparation of (±)-^reo-2-amino-3-(furan-2-yl)-3-hydroxy-l-(pyrrolidin-1yl)propan-l-one hydrochloride Compound 201.

/m//5-5-(Furan-2-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1 -yDmethanone BLE 04136D.

BLE 04136D was prepared in accordance with method D using 2furaldehyde (0.449 mL, 5.42 mmol), KOH (0.276 mg, 4.92 mmol) in methanol (5 mL) and 2-isocyano-l-(pyrrolidin-l-yl)ethanone BLE 04098 (0.75 g, 5.42 mmol). After work-up the residue was purified by column chromatography (SiO₂, cyclohexane:EtOAc = 100:0 to 0:100) to led, after evaporation, to trans-

5-(furan-2-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin- l-yl)methanone BLE 04138D (0.742 g, 58.5 % yield) as a pale yellow oil.

(+)-trans

BLE 04136D

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MW: 234.25; Yield: 58.5 %; Pale Yellow Oil.

^!H-NMR (CDC1₃,0): 1.80-2.10 (m, 4H, CH₂), 3.47-3.60 (m, 3H, CH₂N), 3.934.03 (m, 1H, CH₂N), 4.94 (dd, 1H, J= 7.4 Hz, 7 = 2.2 Hz, CH-N), 6.14 (d, 1H, 7= 7.4 Hz, CH-O), 6.37 (dd, 1H, 7= 3.3 Hz, 7= 1.8 Hz, CH=C), 6.47 (d, 1H, 7 = 3.3 Hz, CH=C), 6.92 (d, 1H, 7= 2.2 Hz, O-CH=N), 7.44 (t, 1H, 7= 1.6 Hz, OCH=C).

(±)-/^reo-2-Amino-3-(furan-2-yl)-3-hydroxy-1 -(pyrrolidin-1 -yppropan-1 -one hydrochloride Compound 201.

Compound 201 was prepared following method E with /r«zw-(5-(furan-2yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-l-yl)methanone BLE 04136D (0.30 g,

1.28 mmol), hydrochloric acid 37 % (0.3 mL) and methanol (10 mL). After overnight at RT and work-up (±)-/7zre(9-2-amino-3-(furan-2-yl)-3-hydroxy-l(pyrrolidin-l-yl)propan-l-one hydrochloride (0.22 g, 66 % yield) was obtained as a pale brown solid.

oh o

HCI (±)

Compound 201

MW: 260.72; Yield: 66 %; Pale Brown Solid; Mp (°C): 159.8

Ή-NMR (CD₃OD,0): 1.62-1.95 (m, 4H, 2xCH₂), 2.72-2.85 (m, 1H, CH₂N),

3.22-3.35 (m, 1H, CH₂N), 3.38-3.55 (m, 2H, CH₂N), 4.35 (d, 1H, 7= 8.5 Hz, CH-N), 4.91 (d, 1H, 7= 8.5 Hz, CH-O), 6.45 (m, 2H, ArH), 7.52-7.57 (m, 1H, ArH).

¹³C-NMR (CD₃OD,0): 24.9, 26.9, 47.4, 47.6, 57.4, 67.8, 109.9, 111.9, 144.4, 153.0,166.0.

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MS-ESI m/z (% rel. Int.): 225.1 ([MH]⁺, 18), 207.1 (100).

HPLC: Method A, detection UV 254 nm, Compound 201 RT = 2.87 min, peak area 92.0 %.

Preparation of (-)-(2R,3_<S)-2-amino-3-hydroxy-3-(pyridin-4-yl)-l-(pyrrolidin-1yl)propan-l-one dihydrochloride Compound 203 and (+)-(2_tS',3R)-2-amino-3hydroxy-3 -ip yridin-4-yI)-1 -(pyrrolidin-1 -vDpropan-1 -one dihydrochloride Compound 204.

Extraction of the free base of Compound 22:

(±)-iAreo-2-Amino-3-hydroxy-3-(pyridin-4-yl)-1 -(pyrrolidin- l-yl)propan1-one dihydrochloride Compound 22 (350 mg, 1.14 mmol) was dissolved in 20 mL of a K₂CO₃ (10 %) solution and the aqueous mixture was then saturated with NaCl. The aqueous phase was extracted by a mixture CH₂Cl₂:2-PrOH = 9:1 (6x15 mL). The organic phase was dried over MgSO₄ and evaporated to afford 226 mg (85 % yield) of the free base of Compound 22.

Analytical chiral separation:

pL of a 1 mg/mL solution of Compound 22 were injected on Chiralpak AD: flow-rate = 1 mL/min, temperature = 25 °C, mobile phase: hexane:ethanol = 1:1, detection by UV at 220 nm and by polarimeter, Rt (-) = 8.20 min, Rt (+) = 10.61 min, k (-) = 1.72, k (+) = 2.51, a = 1.47 and resolution Rs = 3.08.

Semi-preparative chiral separation:

A solution of 100 mg/mL was prepared and 10 pL of this solution were injected every 4.5 min on Chiralpak AD, flow-rate = 1 mL/min, mobile phase hexane:ethanol = 4:6, detection by UV at 254 nm. 135 successive injections were done. The two main fractions were identified by UV and collected in two

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121 different flasks. The solvent was removed in vacuo at 30 °C. The resulting solid was dissolved in 50 mL of CH₂C1₂ and then filtered on a 0.45 pm millipore membrane. After evaporation of CH₂C1₂, the solid was dissolved in 50 mL of methanol and then filtered. The salts were regenerated according to the procedure reported above.

Regeneration of the salt:

After the chiral separation, about 63 mg of each enantiomer of the free base were dissolved in 100 mL of ethanol and 6.7 mL of HCI (0.2 N, 5 eq) were added. The solvent was evaporated, then 50 mL of ethanol were added and then removed in vacuo and the products were dried over P₂O₅ under vacuum overnight. The enantiomeric purity of the products was checked by analytical injection of the regenerated salts: (-)-(2R,3_<y)-2-Amino-3-hydroxy-3-(pyridin-4-yl)-1 -(pyrrolidin-1 -yPpropan-1 one dihydrochloride Compound 203.

OH o

ΝΛ

NH₂ LV

HCI HCI (-)-threo

Compound 203

MW: 308.20; 83 mg obtained; Yield: 23.5 %; White Solid; Mp (°C): 183.5 Enantiomeric excess = 99.3 % a²⁵ _D = - 22.7 (MeOH, c = 0.51).

(+)-(25,3R)-2-Amino-3-hydroxy-3-(pyridin-4-yP-1 -(pyrrolidin-1 -yDpropan-1 one dihydrochloride Compound 204.

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(+)-threo

Compound 204

MW: 308.20; 82 mg obtained; Yield: 23.5 %; White Solid; Mp (°C): 176.9 Enantiomeric excess = 98.5 % a²⁵ _D = + 23.1 (MeOH, c = 1).

Preparation of (+)-//zreo-2-amino-3-hydroxy-l-(pyrrolidin-l-yl)-3-(thiophen-3yDpropan-l-one hydrochloride Compound 205 and (-)-/Z?reo-2-amino-3hydroxy-1 -(pyrrolidin-1 -yP-3-(thiophen-3 -yppropan-1 -one hydrochloride Compound 206.

Extraction of the free base:

(±)-i7zreo-2-amino-3-hydroxy-1 -(pyrrolidin-1 -yl)-3-(thiophen-3yl)propan-l-one hydrochloride Compound 23 (243 mg, 0.88 mmol) was dissolved in 10 mL of a Na₂CO₃ (10 %) solution and the aqueous mixture was then saturated with NaCl. The aqueous phase was extracted by 5 x 15 mL of a mixture CH₂Cl₂:2-PrOH = 9:1. The organic phase was dried over MgSC>4 and evaporated to afford 190 mg (90 %) of the free base of Compound 23. Analytical chiral separation:

pL of a 1 mg/mL solution of Compound 23 were injected on an analytical Chiralpak AD: flow-rate = 1 mL/min, temperature = 25 °C, mobile phase: ethanol, detection by UV at 220 nm and by polarimeter, Rt (+) - 4.98 min, Rt (-) = 6.23 min, k (+) = 0.55, k (-) = 0.93, a = 1.17 and resolution Rs = 3.34.

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Regeneration of the salt:

After the chiral separation, about 70 mg of each enantiomer of the free base were dissolved in 100 mL of ethanol and 3.6 mL of HCI (0.2 N, 2.5 eq) were added. The solvent was evaporated then 50 mL of ethanol were added and then removed in vacuo. The product was dissolved in 2 mL of methanol and 3 mL of ethyl acetate were added. The solvents were removed to give a white solid and then, the solids were dried over P2O5 under vacuum overnight. Semi-preparative chiral separation:

A 175 mg/mL solution of the free base was prepared and 6 pL of this solution were injected every 3 min on an analytical Chiralpak AD, flow-rate = 1 mL/min, mobile phase ethanol, detection by UV at 254 nm. 150 successive injections were done. The two main fractions were identified by UV and collected in two different flasks. The solvent was removed in vacuo at 30 °C. The resulting solid was dissolved in 50 mL of CH₂C1₂ and then filtered on a 0.45 pm millipore membrane. After evaporation of CH₂C1₂, the solid was dissolved in 50 mL of methanol and then filtered. The salt was regenerated according to the procedure reported above.

The enantiomeric purity of the products was checked by analytical injection of the regenerated salts:

(+)-rizreo-2-Amino-3 -hydroxy-1 -(pyrrolidin-1 -yP-3-(thiophen-3 -vPpropan-1 one hydrochloride Compound 205.

Compound 205

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MW: 276.78; 83 mg obtained; Yield: 34 %; White Solid; Mp (°C): too hygroscopic.

Enantiomeric excess = 99.5 % oc²⁵ _D = + 20.4 (MeOH, c = 0.5).

(-)-f//reo-2-Amino-3-hydroxy-1 -(pyrrolidin-1 -yl)-3 -(thiophen-3-yPpropan-1 one hydrochloride Compound 206.

OH O nh₂

HCI (-)-fhreo

Compound 206

MW: 276.78; 77 mg obtained; Yield: 32 %; White Solid; Mp (°C): too hygroscopic.

Enantiomeric excess = 99.0 % a²⁵ _D = - 20.0 (MeOH, c = 0.52).

Preparation of (-)-/Areo-2-amino-3-hydroxy-3-(pyridin-3-yP-l-(pyrrolidin-lyl)propan-l-one dihydrochloride Compound 207 and (+)-/Z?reo-2-amino-3hydroxy-3-(pyridin-3 -yl)-1 -(pyrrolidin-1 -yPpropan-1 -one dihydrochloride Compound 208.

Semi-preparative separation was performed on Chiralpak AD (250x10 mm): The semi-preparative chiral separation needed three steps :

First step A: A 80 mg/mL solution of (±)-/Z/reo-2-amino-3-hydroxy-3(pyridin-3-yl)-l-(pyrrolidin-l-yl)propan-l-one Compound 20 free base (220 mg) racemate was prepared and 200 pL of this solution were injected every 8

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125 min on Chiralpak AD, flow-rate = 4 mL/min, mobile phase: ethanol, detection by UV at 290 nm. Two main fractions were collected after 13 successive injections:

• 1A containing about 61 mg of (-) enantiomer Compound 207 free base with ee > 97 %.

• 2A containing about 135 mg of mixture +/- enantiomers in a 74/26 ratio.

Second step B: A 30 mg/mL solution of fraction 2A was prepared and 100 pL of this solution were injected every 6 min on Chiralpak AD, flow-rate = 4 mL/min, mobile phase: ethanol, detection by UV at 290 nm. Two main fractions were collected after 45 successive injections:

• IB containing about 27 mg of (-) enantiomer Compound 207 free base with ee > 97 %.

• 2B containing about 105 mg of mixture (+)/(-) in a 93/7 ratio.

Third step C: A 15 mg/mL solution of fraction 2B was prepared and 250 pL of this solution were injected every 6 min on Chiralpak AD, flow-rate = 4 mL/min, mobile phase: ethanol, detection by UV at 254 nm. Two main fractions were collected after 28 successive injections:

• 1C containing about 7 mg of (-) enantiomer Compound 207 free base with ee > 97 %.

• 2C containing about 89 mg of (+) enantiomer with ee > 97 %.

Fractions 1A, IB and 1C of (-) enantiomer Compound 207 free base were mixed together. Fraction 2C of (+) enantiomer Compound 208 free base was taken alone. For the both enantiomers, the solvent was removed in vacuo at 30 °C. The resulting solid was dissolved in 50 mL of CH2Q2 and then filtered on a 0.45 pm millipore membrane. After evaporation of CH₂C1₂, the solid was dissolved in 50 mL of methanol and then filtered. The salt was regenerated

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126 according to the procedure reported below. The intermediate fraction collected contains 25 mg of a mixture of the both enantiomers in 50/50 (+)/(-) ratio and some impurities.

Regeneration of the salt (dihydrochloride):

After the chiral separation, about 90-95 mg of each enantiomers of the free base were dissolved in 100 mL of ethanol and 10 mL of HCI (0.2 N, 5 eq) were added. The solvent was evaporated and 50 mL of ethanol were added and then removed in vacuo. The product was dissolved in 1 mL of methanol and 5 mL of ethyl acetate were added to precipitate the salt. The solvents were removed to give a white solid and then, the solids were dried over P2O5 under vacuum overnight.

The enantiomeric purity of the products was checked by analytical HPLC injection of the regenerated dihydrochloride salts:

(-)-^reo-2-Amino-3-hydroxv-3-(pyridin-3-yl)-1 -(pyrrolidin-1 -vDpropan-1 -one dihydrochloride Compound 207.

Compound 207

MW: 308.20; 124 mg obtained; Yield: 43 %; White Solid; Mp (°C): 120.4 Enantiomeric excess = 97.8 % measured by HPLC at 220 nm (Chiralpak AD) RT = 6.24 min, eluent ethanol, flow 1 mL/min.

a²⁵ _D = -15.9 (MeOH, c = 1).

(+)-/Areo-2-Amino-3 -hydroxy-3-(pvridin-3-yl)-1 -(pyrrolidin-1 -yl)propan-1 -one dihydrochloride Compound 208.

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Compound 208

MW: 308.20; 117 mg obtained; Yield: 40.5 %; White Solid; Mp (°C): 120.1 Enantiomeric excess = 98.0 % measured by HPLC at 220 nm (Chiralpak AD) RT = 7.39 min, eluent ethanol, flow 1 mL/min.

a²⁵o = + 15.8 (MeOH, c= 1).

Preparation of (+)-riireo-2-amino-3-hydroxy-3-(2-iodophenyl)-l-(pyrrolidin-lyl)propan-l-one hydrochloride Compound 209.

Zmw5'-(4,5-Dihydro-5-(2-iodophenyl)oxazol-4-yl)(pyrrolidin-l-yl)methanone VIB 01090A.

VIB 01090A was prepared in accordance with method D using 2isocyano-1-(pyrrolidin-l-yl)ethanone SLA 09100 (327.9 mg, 2.155 mmol), potassium hydroxide (121 mg, 2.155 mmol) in methanol (2.2 mL) and 2-iodobenzaldehyde (500 mg, 2.155 mmol). The solution was stirred for 3 h at 0 °C. After work-up the crude product was purified by column chromatography (florisil, EtOAc:MeOH = 9:1) to obtain after evaporation iran5-(4,5-dihydro-5(2-iodophenyl)oxazol-4-yl)(pyrrolidin-l-yl)methanone VIB 01090A as a yellow oil (364 mg, 44 % yield).

(±)-trans

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VIB 01090A

MW: 384.22; Yield: 44 %; Yellow Oil.

R_f: 0.51 (EtOAc:MeOH = 9:1).

Ή-NMR (CDC1₃, δ): 1.85-2.07 (m, 4H, 2xCH₂), 3.50-3.62 (m, 3H, CH₂), 3.78-

3.90 (m, 1H, CH₂), 4.57 (dd, 1H, J= 5.6 Hz, J= 1.9 Hz, CH-N), 6.19 (d, 1H, J = 5.6 Hz, CH-O), 7.05 (dt, 1H, J= 7.7 Hz, J= 1.6 Hz, ArH), 7.15 (d, 1H, J=

1.9 Hz, HC=N), 7.27 (dd, 1H, J= 7.9 Hz, .7=1.6 Hz, ArH), 7.39 (t, 1H, J= 7.3 Hz, ArH), 7.87 (d, 1H, J= 7.8 Hz, ArH).

¹³C-NMR (CDC1₃, δ): 24.3, 26.0, 46.3, 46.6, 74.8, 84.4, 95.0, 126.5, 128.4,

129.9, 139.8, 142.2, 155.7,167.1.

(±)-/Areo-2-Amino-3 -hydroxy-3-(2-iodophenyl)-1 -(pyrrolidin-1 -yBpropan-1 one hydrochloride Compound 209.

Compound 209 was prepared following method E with trans-(4,5dihydro-5-(2-iodophenyl)oxazol-4-yl)(pyrrolidin-1 -yl)methanone VIB 01090A (0.345 g, 0.89 mmol), HCI 37 % (0.22 mL) and methanol (4 mL). After heating at 50 °C for 3 h and work-up, a trituration with EtOAc followed by filtration and drying afforded to (±)-/Areo-2-amino-3-hydroxy-3-(2-iodophenyl)-l(pyrrolidin-l-yl)propan-l-one hydrochloride Compound 209 as a white solid (287 mg, 74 % yield).

I OH O

(±)

Compound 209

MW: 396.65; Yield: 74 %; White Solid; Mp (°C): 164.0

Ή-NMR (CD₃OD, δ): 1.47-1.90 (m, 4H, 2xCH₂), 1.95-2.10 (m, 1H, CH₂),

3.25-3.55 (m, 3H, CH₂), 4.23 (d, 1H, J= 9.0 Hz, CH'N), 5.20 (d, 1H, J= 9.0

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Hz, CH-O), 7.11 (t, 1H, 7.4 Hz, ArH), 7.50 (t, 1H, J= 7.5 Hz, ArH), 7.78 (d, 2H, J= 7.9 Hz, ArH), 7.88 (d, 2H, J= 7.7 Hz, ArH).

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129 ¹³C-NMR (CD₃OD, δ): 24.8, 26.8, 47.8,48.1, 58.6, 75.8, 98.3, 130.1, 130.8,

131.8,141.1, 143.8, 165.6.

MS-ESI m/z (% rel. Int.): 360.9 ([MH]⁺, 100), 342.9 (40).

HPLC: Method A, detection UV 254 nm, Compound 209 RT = 3.88 min, peak area 97.8 %.

Preparation of (±)-fZ?reo-2-amino-3-hydroxy-3-hydroxy-3-(4-iodophenyl)-1 (pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 210.

frflHs-(4,5-Dihydro-5-(4-iodophenyl)oxazol-4-yl)(pyrrolidin-l-yl)methanone VIB 01090B.

VIB 01090B was prepared in accordance with method D using 2isocyano-1-(pyrrolidin-l-yl)ethanone SLA 09100 (327.9 mg, 2.155 mmol), potassium hydroxide (121 mg, 2.155 mmol) in methanol (2.2 mL) and 4iodobenzaldehyde (500 mg, 2.155 mmol). The solution was stirred for 3 h at 0 °C. After work-up, the crude product was washed in a minimum amount of MeOH and filtered to obtain after drying trans-(A,5-dihydro-5-(4iodophenyl)oxazol-4-yl)(pyrrolidin-l-yl)methanone VIB 01090B as a white solid (0.377 g, 52 % yield).

(±)-frans

VIB 01090B

MW: 384.22; Yield: 52 %; White Solid; Mp (°C): 115.1

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130 ’H-NMR (CDC1₃, δ): 1.75-2.10 (m, 4H, 2xCH₂), 3.35-3.60 (m, 3H, CH₂), 3.884.02 (m, 1H, CH₂), 4.53 (dd, 1H, J= 7.7 Hz, J= 1.9 Hz, CH-N), 6.09 (d, 1H, J = 7.7 Hz, CH-O), 6.92-7.11 (m, 3H, 2xArH & CH=N), 7.69-7.70 (dd, 2H, J=

8.4 Hz, J= 1.7 Hz, ArH).

¹³C-NMR (CDC1₃, δ): 24.1, 26.0, 46.2, 46.4, 75.7, 80.8, 94.0, 127.6 (2xC),

137.9 (2xC), 139.4, 155.2, 166.4.

(±)-/Areo-2-Amino-3-hydroxy-3-hydroxy-3-(4-iodophenyl)-l -(pyrrolidin-1 yDpropan-l-one dihydrochloride Compound 210.

Compound 210 was prepared following method E with trans-(4,5dihydro-5-(4-iodophenyl)oxazol-4-yl)(pyrrolidin-1 -yl)methanone VIB 01090B (0.345 g, 0.89 mmol), hydrochloric acid 37 % (0.24 mL) and methanol (4.4 mL). After heating at 50 °C for 3 h and work-up, a trituration with EtOAc followed by filtration and drying afforded to (±)-/Areo-2-amino-3-hydroxy-3-(4iodophenyl)-1-(pyrrolidin-l-yl)propan-l-one hydrochloride Compound 210 as a white solid (247.5 mg, 64 % yield).

Compound 210

MW: 396.65; Yield: 64 %; White Solid; Mp (°C): 184.4

Ή-NMR (CD₃OD, δ): 1.35-1.9 (m, 4H, 2xCH₂), 2.20-2.33 (m, 1H, CH₂), 3.183.40 (m, 3H, CH₂), 4.11 (d, 1H, J= 8.9 Hz, CH’N), 4.82 (d, 1H, J- 8.9 Hz, CH-O), 7.22 (d, 2H, J= 8.2 Hz, ArH), 7.76 (d, 2H, J= 8.2 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 24.8,26.6, 47.3,47.7, 59.2, 73.6, 95.1, 129.8 (2xC),

138.9 (2xC), 140.6, 166.1.

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MS-ESI m/z (% rel. Int.): 360.9 ([MH]⁺, 100), 342.9 (85).

HPLC: Method A, detection UV 254 nm, Compound 210 RT = 4.08 min, peak area 96.8%.

Preparation of (±)-//?rgo-2-amino-3-hydroxy-3-(3-iodophenyl)-1 -(pyrrolidin-1 yl)propan-l-one hydrochloride Compound 211.

/Fa770^,-(4,5-dihydro-5-(3-iodophenyl)oxazol-4-yl)(pyrrolidin-l-yl)methanone SLA 09104.

To a stirred and cooled (0 °C) solution of KOH (0.144 g, 2.57 mmol) in methanol (10 mL) was added 3-iodobenzaldehyde (0.500 mg, 2.15 mmol) and l-isocyano-3-(pyrrolidin-l-yl)propan-2-one (0.295 g, 2.13 mmol). The solution was stirred 24 h with continued cooling and then concentrated. Water (50 mL) was added and the solution was extracted with EtOAc (3 x 50 mL). The organic layer was washed with brine (50 mL), dried over MgSO4, filtered and evaporated. /ra7Z5-(4,5-Dihydro-5-(3-iodophenyl)oxazol-4-yl)(pyrrolidin-lyl)methanone SLA 09104 was obtained (0.63 g, 80 % yield) as a pale yellow solid.

(±)-trans

SLA 09104

MW: 370.19; Yield: 80 %; Pale Yellow Solid.

Ή-NMR (CDC1₃, δ): 1.80-2.08 (m, 4H, 2xCH₂), 3.42-3.58 (m, 3H, 1.5xCH₂),

3.90-3.96 (m, 1H, 0.5xCH₂), 4.56 (dd, 1H, J= 7.7 Hz, J= 2.2 Hz, CH-N), 6.10

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132 (d, 1H, J= 7.7 Hz, CH-O), 7.01 (d, 1H, J= 2.2 Hz, CH=N), 7.11 (t, 1H, J= 6.8 Hz, ArH), 7.29 (m, 1H, ArH), 7.66 (m, 2H, ArH).

¹³C-NMR (CDCI3, δ): 24.2, 26.0, 46.5,46.6, 75.8, 80.4, 94.7, 125.0, 130.6,

134.6,137.5, 142.0, 155.2, 166.3.

(±)-/Areo-2-Amino-3-hydroxy-3-(3-iodophenyP-1 -(pyrrolidin-1 -yppropan-1 one hydrochloride Compound 211.

trans-(4,5-Dihydro-5-(3-iodophenyl)oxazol-4-yl)(pyrrolidin-lypmethanone SLA 09104 (0.620 g, 1.67 mmol) was dissolved in methanol (5 mL). The solution was stirred at room temperature and a solution of HCI (37 %, 1 mL) was added via syringe and the mixture was stirred at 50 °C for 6 h. The mixture was concentrated and triturated with EtOAc. After filtration and drying (±)-/Areo-2-amino-3-hydroxy-3-(3-iodophenyl)-1-(pyrrolidin-1 -yl)propan-1 -one hydrochloride Compound 211 was obtained (586 mg, 88 % yield) as a white solid.

Compound 211

MW: 396.65; Yield: 88 %; White Solid; Mp (°C): 183.7

Ή-NMR (CD₃OD, δ): 1.35-1.50 (m, 1H, 0.5xCH₂), 1.58-1.82 (m, 3H, 1.5xCH₂), 2.08-2.18 (m, 1H, 0.5xCH₂), 3.21-3.45 (m, 4H, 2xCH₂), 4.09 (d, 1H, J= 9.1 Hz, CH-N), 4.80 (d, 1H, J= 9.2 Hz, CH-O), 7.19 (t, 1H, J= 7.1 Hz, ArH), 7.49 (d, 1H, J= 7.6 Hz, ArH), 7.74 (m, 2H, ArH).

MS-ESI m/z (% rel. Int.): 360.9 ([MH]⁺, 100), 342.9 (40).

HPLC: Method A, detection UV 254 nm, Compound 211 RT = 4.07 min, peak area 93.4 %.

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Preparation of (±)-//zreo-2-amino-3-hydroxy-1 -(4-methylpiperazin-1 -yl)-3(pyridin-4-yDpropan-l-one trihydrochloride Compound 212.

2-Isocyano-l-(4-methyl-piperazin-l-yl)-ethanone VIB 01128.

Prepared in accordance with Method B with methyl isocyanoacetate (1.0 g, 10.09 mmol) and A-methylpiperazine (1.37 mL, 15.14 mmol). The reaction mixture was stirred overnight at RT and then concentrated. The residue was dissolved in dichloromethane (50 mL) and evaporated. The residue was coevaporated three times with a mixture of CH₂C1₂:cyclohexane = 50:50 (3x10 mL). After drying 2-isocyano~l-(4-methyl-piperazin-l-yl)-ethanone VIB 01128 was obtained as yellow solid (1.67 g, 99 % yield).

VIB 01128

MW: 167.21; Yield: 99 %; Yellow Solid; Mp (°C) = 106.0

Ή NMR (CDC1₃, δ): 2.32 (m, 3H, Me) 2.38-2.50 (m, 4H, 2xCH₂), 3.42 (t, 2H, J = 4.7 Hz, CH₂), 3.66 (t, 2H, J= 4.7 Hz, CH₂), 4.30 (s, 2H, CH₂). ¹³C-NMR (CDCI3, δ): 42.4,44.4, 45.5,46.0, 54.3, 54.5,160.8, 161.2.

/mw.y-(4,5-Dihydro-5-(pyridin-4-yl)oxazol-4-yl)(4-methylpiperazin-lyl)methanone VIB 01130.

To a stirred and cooled (0 °C) solution of KOH (0.335 g, 5.98 mmol) in 7 mL MeOH were added successively pyridine-4-carbaldehyde (0.705 g, 6.58 mmol) and 2-isocyano-l-(4-methyl-piperazin-l-yl)-ethanone VIB 01128 (1.00

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134 g, 6.58 mmol). The mixture was stirred at 0 °C until precipitation and concentrated. The mixture was partitioned between EtOAc (20 mL) and H2O (10 mL). The aqueous layer was extracted twice with EtOAc (60 mL). The EtOAc fractions were combined, washed twice with brine (2x10 mL), dried over MgSO₄ and filtered. After evaporation and drying iran5-(4,5-dihydro-5(pyridin-4-yl)oxazol-4-yl)(4-methylpiperazin-l-yl)methanone VIB 01130 was obtained (1.282 g, 78 % yield) as a yellow oil.

° (+)-trans

VIB 01130

MW: 274.32; Yield: 78 %; Yellow Oil.

^!H NMR (CDCI3, δ): 2.32 (m, 3H, Me) 2.35-2.58 (m, 4H, 2xCH₂), 3.50-3.70 (m, 2H, CH₂-N), 3.80-4.00 (m, 2H, CH₂-N), 4.59 (dd, 1H, J= 7.8 Hz, J= 2.2 Hz, CH-N), 6.27 (d, 1H, 7= 7.8 Hz, O-CH), 7.02 (d, 1H, J= 2.2 Hz, OCH=N), 7.23 (d, 2H, 4.7 Hz, ArH), 8.62 (dd, 2H, J= 4.5 Hz, J= 1.6 Hz, ArH).

¹³C-NMR (CDC1₃, δ): 42.6, 45.7, 46.0, 54.6, 55.1, 74.7, 79.6, 120.0 (2xC), 148.6,150.3 (2xC), 154.8,166.0.

MS-ESI m/z (% rel. Int.): 275.2 ([MH]⁺, 40), 190.1 (35), 147.0 (40), 127.0 (100).

HPLC: Method A, detection UV 254 nm, VIB 01130 RT = 0.70 min, peak area

99.9 %.

(±)-/Areo-2-Amino-3-hydroxy-1 -(4-methylpiperazin-1 -yl)-3 -(pyridin-4yl)propan-l-one trihydrochloride Compound 212.

To a solution of Zraw5-(4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)(4methylpiperazin-l-yl)methanone VIB 01130 (1.235 g, 4.50 mmol) in MeOH

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135 (15 mL) was added HCI 37 % (1.6 mL). After heating (50 °C) the mixture for

3.5 h a white solid precipitated and the reaction mixture was concentrated and the crude product was coevaporated twice with ethyl acetate. After trituration with ethyl acetate, filtration and drying, (±)-/Areo-2-amino-3-hydroxy-1-(4methylpiperazin-1 -yl)-3-(pyridin-4-yl)propan-1 -one trihydrochloride Compound 212 (1.62 g, 96 % yield) was obtained as a white solid.

OH o

Compound 212

MW: 373.71; Yield: 96 %; White Solid; Mp (°C): 203.1

Ή-NMR (CD₃OD, δ): 2.95 (s, 3H, Me), 3.10-3.40 (m, 4H, 2xCH₂), 3.48-3.80 (m, 2H, CH₂), 4.19-4.49 (m, 1H, 0.5xCH₂), 4.51-4.79 (m, 1H, 0.5xCH₂), 5.06 (d, 1H, J= 4.3 Hz, CH-NH₂), 5.35-5.63 (m, 1H, CH-O), 8.30 (s broad, 2H, ArH), 8.95 (d, 2H, J= 6.7 Hz, ArH).

MS-ESI m/z (% rel. Int.): 265.1 ([MH]⁺, 5), 248.1 (20), 156.1 (20), 148.0 (100). HPLC: Method A, detection UV 254 nm, Compound 212 RT = 0.68 min, peak area 99.9 %.

Preparation of (±)-7zreo-2-amino-1 -(3,3-difluoropyrrolidin-1 -yl)-3-hydroxy-3 (pyridin-4-yl)propan-l-one dihydrochloride Compound 213.

l-(3,3-Difluoropyrrolidin-l-yl)-2-isocyanoethanone VIB 01158.

To stirred and cooled (0 °C) methyl isocyanoacetate (96 % technical grade, 345 mg, 3.48 mmol) was slowly added 3,3-difluoropyrroIidine hydrochloride (500 mg, 3.48 mmol), triethylamine (487 pL, 3.48 mmol) and

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MeOH (1 mL). The mixture was stirred for 15 h at RT and then concentrated.

The resulting oil was coevaporated twice from EtOAc. 1-(3,3Difluoropyrrolidin-l-yl)-2-isocyanoethanone VIB 01158 was obtained as a yellow oil (305 mg, 60 % yield) and used in the next step without purification.

VIB 01158

MW: 174.17; Yield: 60 %; Yellow Oil.

NMR (CDC1₃, δ): 2.30-2.62 (m, 2H, CH₂) 3.65-3.90 (m, 4H, CH₂), 4.25 (d, 2H, 17.5 Hz, CH₂).

Zrans-(3,3-DifluoropyrroIidin-l-yl)(4,5-dihydro-5-(pyridin-4-yl)oxazol-4vDmethanone VIB 01160.

To a stirred and cooled (0 °C) solution of KOH (0.098 g, 1.75 mmol) in MeOH (2 mL) were added successively pyridine-4-carbaldehyde (0.206 g 1.92 mmol) and 1-(3,3-difluoropyrrolidin-l-yl)-2-isocyanoethanone VIB 01158 (0.305 g, 1.75 mmol). The mixture was stirred at 0 °C for 3 h. After evaporation of MeOH, the mixture was partitioned between EtOAc (50 mL) and H₂O (40 mL). The aqueous layer was extracted with EtOAc (4x50 mL). The fractions were combined, washed twice with brine (2x20 mL), dried over MgSO4 and filtered. After evaporation and drying trans-(3,3-difluoropyrrolidin-l-yl)(4,5dihydro-5-(pyridin-4-yl)oxazol-4-yl)methanone VIB 01160 (0.33 g, 67 % yield) was obtained as a yellow oil.

VIB 01160

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MW: 281.26; Yield: 67 %; Yellow Oil.

Ή NMR (CDC1₃, δ): 2.30-2.60 (m, 2H, CH₂) 3.67-3.99 (m, 3H, 1.5xCH₂), 4.22-

4.59 (m, 2H, CH-N & 0.5xCH₂), 6.17 (d, 1H, J= 7.8 Hz, CH-O), 7.04 (dd, 1H, J= 3.2 Hz, J= 2.5 Hz, HC=N), 7.20-7.38 (m, 2H, ArH), 8.55-8.70 (m, 2H, ArH).

(±)-//zrgo-2-Amino-1 -(3,3-difluoropyrrolidin-1 -yl)-3-hydroxy-3-(pyridin-4yl)propan-l-one dihydrochloride Compound 213.

To a solution of trans-(3,3-difluoropyrrolidin-l-yl)(4,5-dihydro-5(pyridin-4-yl)oxazol-4-yl)methanone VIB 01160 (0.305 g, 1.08 mmol) in methanol (4 mL) was added hydrochloric acid 37 % (395 pL). After heating (50°C) the mixture for 3.5 h the reaction mixture was concentrated and the crude product was coevaporated twice with EtOAc. After trituration with EtOAc, filtration and drying, (±)-/Areo-2-amino-l-(3,3-difluoropyrrolidin-l-yl)-

3-hydroxy-3-(pyridin-4-yl)propan-l-one dihydrochloride Compound 213 (269 mg, 44 % yield) was obtained as a beige solid.

Compound 213

MW: 344.19; Yield: 44 %; Beige Solid; Mp (°C): 182.2

Ή-NMR (CD₃OD, δ): 2.25-2.62 (m, 2H, CH₂), 3.30-4.20 (m, 4H, CH₂), 4.55 (d, 1H, J= 5.5 Hz , 0.35xCH-N), 4.66 (d, 1H, J= 5.5 Hz , 0.65xCH’N), 5.42 (m, 1H, CH-O), 8.23 (d, 2H, 4.8 Hz, ArH), 8.94 (d, 2H, J= 5.9 Hz, ArH).

MS-ESI m/z (% rel. Int.): 272.2 ([MH]⁺, 15), 254.1 (15), 178.1 (20), 148.1 (100), 137.1 (95).

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HPLC: gradient of solvent B:solvent A = 5:95 to 100% solvent B in 7 min. Solvent A was FLO with 0.1 % Et₃N and solvent B was CH₃CN with 0.1 % Et₃N; detection UV 254 nm, Compound 213 RT = 4.70 min, peak area 98.1 %.

Preparation of (2S,3R)~ & (2R,363-2-anrino-l-(6S)-3-fhioropyrrolidin-l-yl)-3hydroxy-3-(pyridin-4-yl)propan-l-one dihydrochlorides Compounds 214.

l-((5)-3--Fluoropyrrolidin-l-yl)-2-isocyanoethanone VIB 01166.

To stirred and cooled (0 °C) methyl isocyanoacetate (96 % technical grade, 1,18 g, 11.9 mmol) was slowly added (0)-(+)-3-fluoropyrrolidine hydrochloride (97%, 1.5 g, 11.9 mmol), triethylamine (1.67 mL, 11.9 mmol) and MeOH (3 mL). The mixture was stirred for 15 h at RT and concentrated. The mixture was stirred for 15 h at RT and concentrated. Water was added (50 mL) and the mixture was extracted with EtOAc (3x50 mL), dried over MgSO₄, filtered and evaporated to obtained crude l-((<S)-3-fluoropyrrolidin-l-yl)-2isocyanoethanone VIB 01166 (1.47 g, 79 % yield) as a brown oil which was used in the next step without purification.

VIB 01166

MW: 156.152; Yield: 79 %; Brown Oil.

Ή NMR (CDC1₃, δ): 1.85-2.50 (m, 2H, CH₂), 3.40-4.35 (m, 6H, 3xCH₂), 5.175.47 (m, 1H, CHF).

MS-ESI m/z (% rel. Int.): 171.1 ([MH⁺+Na], 100), 157.1 ([MH]⁺, 82), 130.1 (95).

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Zmw0'-((5)-3-Fluoropvrrolidin-l-vl)((45,57?)- & 47?,55)-4,5-dihydro-5-(pvridin-4yl)oxazol-4-yl)methanones VIB 01168.

To a stirred and cooled (0 °C) solution of KOH (0.526 g, 9.39 mmol) in MeOH (10 mL) were added successively pyridine-4-carbaldehyde (1.10 g, 10.33 mmol) and l-((5)-3-fluoropyrrolidin-l-yl)-2-isocyanoethanone VIB 01166 (1.47 g, 9.39 mmol). The mixture was stirred at 0 °C to RT for 24 h. After evaporation of MeOH, the mixture was partitioned between EtOAc (40 mL) and H₂O (20 mL). The aqueous layer was extracted with EtOAc (6x40 mL). The EtOAc fractions were combined, washed twice with brine (2x10 mL), dried over MgSO₄, filtered and evaporated. The crude product was purified by column chromatography (florisil, EtOAc:MeOH = 9:1). After evaporation and drying iran5^,-((5)-3-fluoropynolidin-l-yl)((45,57?)- & 47?,55)-4,5-dihydro-5(pyridin-4-yl)oxazol-4-yl)methanones VIB 01168 (590 mg, diastereoisomeric mixture in ratio 1:1, 24 % yield) were obtained as a yellow oil.

trans trans

VIB 01168

MW: 263.27; Yield: 24 %; Yellow Oil.

^JH NMR (CDC1₃, δ): 1.89-2.46 (m, 2H, CH₂) 3.5-4.45 (m, 4H, CH₂), 4.46-4.60 (m, 1H, CH-N), 5.16-5.45 (m, 1H, CH-F), 6.12-6.25 (m, 1H, CH-O), 6.95-7.18 (m, 1H, CH=N), 7.20-7.40 (m, 2H, ArH), 8.50-8.70 (m, 2H, ArH).

(25,37?)- & (27?,35)-2-Amino-l-((5)-3-fluoropyrrolidin-l-yl)-3-hydroxy-3(pyridin-4-yl)propan-l-one dihydrochlorides Compounds 214.

To a solution of ((5)-3-fluoropyrrolidin-l-yl)((45,57?)- & 47?,55)-4,5dihydro-5-(pyridin-4-yl)oxazol-4-yl)methanones VIB 01168 (0.590 g, 2.24

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140 mmol) in methanol (8 mL) was added hydrochloric acid 37 % (686 pL). After heating (50 °C) the mixture for 3.5 h the reaction mixture was concentrated and the crude product was coevaporated twice with EtOAc. After trituration with EtOAc, filtration and drying (2S,3R)- & (2R,3S)-2-amino-1-((5)-3fluoropyrrolidin-1 -yl)-3-hydroxy-3-(pyridin-4-yl)propan-1 -one dihydrochlorides Compounds 214 (474 mg, diastereoisomeric mixture in ratio 1:1, 68 % yield) were obtained as a pale yellow solid.

oh o oh o

Compounds 214

MW: 326.2; Yield: 68 %; Pale Yellow Solid; Mp (°C): 173.1.

Ή-NMR (CD₃OD, δ): 1.82-2.38 (m, 2H, CH₂), 2.80-4.15 (m, 4H, 2xCH₂), 4.35-4.68 (m, 1H, CH N), 5.00-5.50 (m, 2H, CH-O & CH-F), 8.11-8.32 (m, 2H, ArH), 8.82-9.00 (m, 2H, ArH).

MS-ESI m/z (% rel. Int.): 254.2 ([MH]⁺, 15), 237.1 (20), 148.1(100), 137.1 (70).

HPLC: gradient of solvent B: solvent A = 5:95 to 100% solvent B in 7 min. Solvent A was H₂O with 0.1 % Et₃N and solvent B was CH₃CN with 0.1 % Et₃N; detection UV 254 nm, Compounds 214 RT = 4.35 min, peak area 99.0 %.

Preparation of (2S,3R)- & (2R_n3Y)-2-amino-l-((R)-3-fluoropyrrolidin-l-yl)-3hydroxy-3-(pyridin-4-yDpropan-l-one dihydrochlorides Compounds 215.

l-((R)-3-Eluoropyrrolidin-l-yP-2-isocyanoethanone BLE 04170.

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To stirred and cooled (0 °C) methyl isocyanoacetate (96 % technical grade, 0.79 g, 7.96 mmol) was slowly added (R)-(-)-3-fluoropyrrolidine hydrochloride (1.0 g, 7.96 mmol), triethylamine (1.11 mL, 7.96 mmol) and MeOH (2.5 mL). The mixture was stirred for 15 h at RT and concentrated. Water was added (50 mL) and the mixture was extracted with EtOAc (3x50 mL), dried over MgSO4, filtered and evaporated to obtained crude l-((R)-3fluoropyrrolidin-l-yl)-2-isocyanoethanone BLE 04170 as a brown oil (0.96 g, 77 % yield) which was used in the next step without further purification.

BLE 04170

MW: 156.152; Yield: 77 %; Brown Oil.

*H NMR (CDC1₃, δ): 1.85-2.50 (m, 2H, CH₂), 3.40-4.35 (m, 6H, 3xCH₂), 5.175.47 (m, 1H, CHF).

/rans-((R)-3-Fhioropyrrolidin- Ι-νΠΙΗ^Τ?)- & 4R,5S^f)-4,5-dihydro-5-(pyridin-

4-yl)oxazol-4-yl)methanones BLE 04172.

To a stirred and cooled (0 °C) solution of KOH (0.34 g, 6.06 mmol) in MeOH (4.5 mL) were added successively pyridine-4-carbaldehyde (0.71 g, 6.66 mmol) and a solution of BLE 04170 (0.95 g, 6.06 mmol) in MeOH (2.5 mL). The mixture was stirred at 0 °C to RT for 15 h. After evaporation of MeOH, the mixture was partitioned between EtOAc (40 mL) and H₂O (20 mL). The aqueous layer was extracted with EtOAc (3x40 mL). The EtOAc fractions were combined, washed with brine (50 mL), dried over MgSO4, filtered and evaporated. The crude product was purified by column chromatography (florisil, EtOAc:MeOH = 9:1). After evaporation and drying trans-((R)-3fluoropyrrolidin-1 -yl)((4S,5R)- & 4R,55)-4,5-dihydro-5-(pyridin-4-yl)oxazol-4WO 2006/081273

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trans trans

BLE 04172

MW: 263.27; Yield: 41 %; Colorless Oil.

NMR (CDC1₃, δ): 1.S9-2.46 (m, 2H, CH₂) 3.5-4.45 (m, 4H, 2xCH_z), 4.46-

4.60 (m, 1H, CH-N), 5.16-5.45 (m, 1H, CH-F), 6.12-6.25 (m, 1H, CH-O), 6.957.18 (m, 1H, CH=N), 7.20-7.40 (m, 2H, ArH), 8.50-8.70 (m, 2H, ArH).

(25,37?)- & (27?,35)-2-Amino-l-((7?)-3-fluoropyrrolidin-l-yl)-3-hydroxy-3(pyridin-4-yDpropan-l-one dihydrochlorides Compounds 215.

To a solution of 7ra«5^,-((7?)-3-fluoropyrrolidin-l-yl)((45,57?)- & 47?,5S)-

4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)methanones BLE 04172 (0.65 g, 2.47 mmol) in methanol (8 mL) was added hydrochloric acid 37 % (757 pL). After heating (50 °C) the mixture for 3.5 h the reaction mixture was concentrated and the crude product was coevaporated twice with EtOAc. After trituration with EtOAc, filtration and drying (25,37?)- & (27?,36)-2-amino-l-((7?)-3fhioropyrrolidin-1 -yl)-3-hydroxy-3-(pyridin-4-yl)propan- 1-one dihydrochlorides Compounds 215 (544 mg, diastereoisomeric mixture in ratio 1:1, 68 % yield) were obtained as a pale yellow solid.

OH O

Compounds 215

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MW: 326.2; Yield: 68 %; Pale Yellow Solid; Mp (°C): 134.5

Ή-NMR (CD₃OD, δ): 1.82-2.38 (m, 2H, CH₂), 2.80-4.15 (m, 4H, CH₂), 4.35-

4.68 (m, 1H, CH'N), 5.00-5.50 (m, 2H, CH-0 & CHF), 8.11-8.32 (m, 2H, ArH), 8.82-9.00 (m, 2H, ArH).

Preparation of A-((+)-/Areo-1 -hydroxy-3-oxo-1 -(pyridin-4-yl)-3-(pyrrolidin-1 yl)propan-2-yl)formamide hydrochloride Compound 216.

trans-(4,5-Dihydro-5-(pyridin-4-yl)oxazol-4-yl)(pyrrolidin-1 yl)methanone Compound 19 (0.200 g, 0.81 mmol) was dissolved in methanol (5 mL). Dowex 50WX8-200 (0.5 mL, washed beforehand by a 0.5 M solution of HCI then water) was added at RT. The mixture was stirred for 1 h at 50 °C and after cooling was filtered off. MeOH was evaporated and the residue was dried to give 150 mg of a pasty product (150 mg, 61.0 % yield). The free base form was dissolved in a minimum of a mixture of EtOAc:MeOH = 95:5 and a 0.4 M solution of HCI in ether (1.3 mL, 0.52 mmol) was added at + 4 °C. After evaporation of solvents, the product was precipitated with diethyl ether, filtered and dried to obtain 7V-((±)-i/zreo-l-hydroxy-3-oxo-l-(pyridin-4-yl)-3(pyrrolidin-l-yl)propan-2-yl)formamide hydrochloride Compound 216 as a beige solid (100 mg, 41.0 % yield).

OH 0

II T £ ,NH OHC

HCI (±)

Compound 216

MW: 299.75; Yield: 41.0 %; Beige Solid; Mp (°C): 203.1

Ry: 0.35 (CH₂Cl₂:MeOH = 90:10) free base.

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144 ³H-NMR (CD₃OD, δ): 1.87-2.00 (m, 4H, 2xCH₂), 3.41-3.46 (m, 2H, CH₂_N),

3.62-3.66 (m, 2H, CH₂-N), 5.18 (d, 1H, J= 3.6 Hz, CH-N), 5.40 (d, 1H, J= 3.8 Hz, CH-O), 7.99 (s, 1H, CH=O), 8.16 (d, 2H, J= 5.8 Hz, ArH), 8.81 (d, 2H, J=

5.5 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 25.0, 27.0, 47.5,48.3, 55.6, 72.4, 126.6 (2xC), 142.1 (2xC), 163.4, 164.1, 168.5.

MS-ESI m/z (% rel. Int.): 264.1 ([MH]⁺, 10), 148.0 (100).

HPLC: Method A, detection UV 254 nm, Compound 216 RT = 1.30 min, peak area 98.0 %.

Preparation of 7V-((±)-///reo-2-acetoxy-2-pyridin-4-yl-1-(pyrrolidine-1carbonyl)-ethyl)-formamide hydrochloride Compound 217,

7V-((±)-i/zreo-1 -hydroxy-3-oxo-1 -(pyridin-4-yl)-3-(pyrrolidin-1 -yl)propan-

2-yl)formamide hydrochloride Compound 216 (0.220 g, 0.80 mmol) was dissolved in CH₂C1₂ (10 mL) with triethylamine (280 pL, 2 mmol) at 0 °C. Then acetic anhydride (160 pL, 1.6 mmol) was added slowly and the mixture was stirred for 72 h at RT. The solvent was evaporated and the residue was dried under vacuum. After column chromatography (SiO₂, EtOAc:MeOH = 90:10) to give a pasty product (100 mg, 41% yield). The product obtained was dissolved in MeOH (10 mL) and a solution of HCI (0.4 Μ, 1 mL) in Et₂O was added at 0 °C. Evaporation of the volatiles led to acetic acid 7V-((±)-/7/reo-2-acetoxy-2pyridin-4-yl-1 -(pyrrolidine-1 -carbonyl)-ethyl)-formamide hydrochloride Compound 217 was obtained as a white solid (110 mg, 40 % yield).

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Compound 217

MW: 341.79; Yield: 40.0 %; White Solid; Mp (°C): 173.9.

R_z: 0.25 (EtOAc:MeOH = 90:10, free base).

^-NMR (CD₃OD,b): 1.80-2.00 (m, 4H, 2xCH₂), 2.19 (s, 3H, CH₃) 3.29-3.64 (m, 4H, 2xCH₂-N), 5.34 (s, 1H, N-CH), 6.44 (s, 1H, O-CH), 8.01 (s, 1H, CHO), 8.16 (s, 2H, ArH), 8.85 (s, 2H, ArH).

¹³C-NMR (CD₃OD, δ): 20.5,25.0,27.0,47.6,47.7, 53.9, 73.4, 126.7 (2xC),

142.8 (2xC), 144.3, 163.3, 166.9, 170.9.

MS-ESI m/z (% rel. Int.): 306.1 ([MH]⁺, 10), 261.1 (100).

HPLC: Method A, detection UV 254 nm, Compound 217 RT = 0.90 min, peak area 97.0 %.

Preparation of ter/-butyl (±)-/Areo-l-hydroxy-3-oxo-l-(pyridin-4-yl)-3(pyrrolidin-l-yl)propan-2-ylcarbamate hydrochloride Compound 218.

(±)-^reo-2-Amino-3-hydroxy-3-(pyridin-4-yl)-l-(pyrrolidin-l-yl)propan1-one dihydrochloride Compound 22 (2.90 g, 9.3 mmol) was dissolved in CH₂C1₂ (250 mL) with Et₃N (4.3 mL, 30.7 mmol) at 0 °C. Di-/er/-butyl dicarbonate (2.45 g, 11.2 mmol) in CH₂C1₂ (50 mL) was added slowly and the mixture was stirred for 15 h at RT. Brine (50 mL) was added and the product was extracted with CH₂C1₂. After drying over MgSCL and filtration, CH₂C1₂ was evaporated and the residue was dried in vacuum. After column chromatography (SiO₂, EtOAc:MeOH = 90:10), teri-butyl (±)-/Areo-l-hydroxy-3-oxo-l(pyridin-4-yl)-3-(pyrrolidin-l-yl)propan-2-ylcarbamate TTA 08100 was

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146 obtained as a beige solid (2.00 g, 64 % yield). A Sample of TTA 08100 (55 mg) was dissolved in CH2CI2 (1 mL) and Et₂O (30 mL) and a solution of HCI (0.1 M, 2 mL) in Et₂O was added at 0 °C. Evaporation of the volatiles led to terZ-butyl (ft)-threo-1 -hydroxy-3-oxo-1 -(pyridin-4-yl)-3-(pyrrolidin-1 yl)propan-2-ylcarbamate hydrochloride Compound 218 (50 mg, 52% yield) as a white solid.

OH 0

Compound 218

MW: 371.86; Yield: 52.0 %; White Solid; Mp (°C): 141.2.

R_z: 0.30 (EtOAc:MeOH - 90:10, free base).

Ή-NMR (CD3OD, δ): 1.33 (s, 9H, 3xCH₃) 1.91-2.00 (m, 4H, 2xCH₂), 3.44O-CH), 8.16 (s, 2H, ArH), 8.83 (s, 2H, ArH).

¹³C-NMR (CD₃OD, δ): 25.0, 27.1, 28.5 (3xC), 47.5, 48.0, 58.3, 72.8, 81.0,

126.7 (2xC), 142.0 (2xC), 157.3, 164.6, 169.5.

MS-ESI m/z (% rel. Int.): 336.1 ([MH]⁺, 20), 219.1 (100).

HPLC: Method A, detection UV 254 nm, Compound 218 RT = 3.8 min, peak area 98.0 %.

Preparation of (±)-eTT/Aro-2-amino-3 -hydroxy-3-(pyridin-4-yl)-1 -(pyrrolidin-1 yl)propan-l-one dihydrochloride Compound 219.

terZ-Butyl (ft)-threo-1 -hydroxy-3 -oxo-1 -(pyridin-4-yl)-3-(pyrrolidin-1 yl)propan-2-ylcarbamate VIB 01080.

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To a solution of (±)-iAreo-2-amino-3-hydroxy-3-(pyridin-4-yl)-l(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 22 (3.25 g, 10.54 mmol) in CH2CI2 (250 mL) in a 500 mL round bottom flask equipped with a magnetic stirrer and under nitrogen atmosphere was added via syringe at 0 °C triethylamine (4.69 mL, 33.73 mmol). A solution of di-teri-butyldicarbonate (2.76 g, 12.65 mmol) in CH₂C1₂ (75 mL) was added at 0 °C dropwise via a dropping funnel. The reaction mixture was abandoned at 0° C for 2 h then at RT overnight. A solution of brine (130 mL) was added and the solution was extracted with CH₂C1₂ (3 x 75 mL), dried over MgSO₄, filtered and evaporated. The crude product was purified by column chromatography (SiO₂, EtOAc:MeOH = 90:10). After evaporation to dryness of the combined fractions, teri-butyl (±)-threo-1 -hydroxy-3-oxo-1 -(pyridin-4-yl)-3-(pyrrolidin-1 yl)propan-2-ylcarbamate VIB 01080 (2.79 g, 79 % yield) was obtained as a yellow solid.

OH 0

VIB 01080

MW: 335.4; Yield: 79 %; Yellow Solid; Mp (°C): 160.5

R/ 0.31 (EtOAc:MeOH = 90:10).

^JH-NMR (CDCI3, δ: 1.26 (s, 9H, 3xCH₃), 1.71-2.00 (m, 4H, 2xCH₂), 3.25-3.60 (m, 4H, 2xCH₂N), 4.61 (dd, 1H, J= 9.8 Hz, 2.6 Hz, N-CH), 4.96 (s, 1H, OH), 5.08 (d, 1H, J= 2.6 Hz, NH), 5.47 (d, 1H, J= 9.8 Hz, O-CH), 7.35 (d, 2H, J= 5.7 Hz, ArH), 8.58 (d, 2H, 5.4 Hz, ArH).

¹³C-NMR (CDC1₃, δ): 24.0 (2xC), 25.9 (2xC), 28.1 (3xC), 46.1,46.7, 56.1,

60.4, 72.8, 80.2, 121.4 (2xC), 148.7, 149.5 (2xC), 155.6, 169.4.

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MS-ESI m/z (% rel. Int.): 336.1 ([MH]⁺, 45), 280 (18), 219 (100); 148 (38).

tert-Butyl (±)-1,3 -dioxo-3-(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)propan-2ylcarbamate TTA 08120.

To a solution of tert-butyl (±)-//zreo-l-hydroxy-3-oxo-l-(pyridin-4-yl)-3(pyrrolidin-l-yl)propan-2-ylcarbamate VIB 01080 (3.92 g, 11.70 mmol) in CH2CI2 (320 mL) in a 500 mL round bottom flask equipped with a magnetic stirrer and under nitrogen atmosphere was added slowly Dess-Martin periodinane (4.96 g, 11.70 mmol) at RT. The reaction mixture was stirred at RT for 0.5 h and CH2Q2, washed with a mixture of saturated sodium bicarbonate (100 mL), 1 M sodium thiosulfate (50 mL), brine (50 mL) and dried over MgSO₄, filtered and evaporated. Diethyl ether (250 mL) was added and the precipitate was filtered off. After evaporation of the filtrate, the crude product was purified by column chromatography (S1O2, CH₂C12:EtOAc = 4:6). After evaporation to dryness of the combined fractions tert-butyl (±)-l,3-dioxo-3(pyridin-4-yl)-l-(pyrrolidin-l-yl)propan-2-ylcarbamate TTA 08120 (3.0 g, 77 % yield) was obtained as a white solid.

TTA 08120

MW: 333.38; Yield: 77 %; White Solid; Mp (°C): 125.4

R_y: 0.25 (CH₂Cl₂:EtOAc = 4:6).

Ή-NMR (CDCIs, δ): 1.36 (s, 9H, 3xCH_s), 1.86-2.00 (m, 4H, 2xCH₂), 3.47-3.71 (m, 4H, 2xCH₂N), 5.62 (d, 1H, J= 7.4 Hz, N-CH), 6.07 (d, 1H, J= 7.4 Hz, NH), 7.80 (dd, 2H, J= 4.6 Hz, J= 1.3 Hz, ArH), 8.80 (dd, 2H, J= 4.5 Hz, J=

1.5 Hz, ArH).

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149 ¹³C-NMR (CDC1₃, δ): 24.0, 26.0, 28.1 (3xC), 46.8, 47.0, 61.0, 80.9, 121.5 (2xC), 141.4, 150.8 (2xC), 155.1, 163.8,194.3.

MS-ESI m/z (% rel. Int.): 334.1 ([MH]⁺, 10), 173.1 (30), 129.1 (100). tert-Butyl (+)-ery^ro-3 -hydroxy-1 -oxo-3-(pyridin-4-yl)-1 -(pyrrolidin-1 yl)propan-2-ylcarbamate TTA 08124P.

To a solution of tert-butyl (±)-l,3-dioxo-3-(pyridin-4-yl)-l-(pyrrolidin-1yl)propan-2-ylcarbamate TTA 08120 (2.28 g, 6.80 mmol) in MeOH (50 mL) in a 250 mL round bottom flask equipped with a magnetic stirrer and under nitrogen atmosphere was added slowly sodium borohydride (285 mg, 7.50 mmol) at RT. The reaction mixture was stirred at RT for 0.5 h and was cooled at 4 °C. A solution of 2 M NaOH (10 mL) was added and MeOH was evaporated at 30 °C. EtOAc (300 mL) was added and the organic phase was washed with brine (20 mL), dried over MgSO4, filtered and evaporated to give a crude compound (ratio erythro:threo = 80:20 estimated by Ή NMR 300 MHz) TTA 08124 (2.1 g, 92 % yield). The crude product was recrystallized in EtOAc and after filtration and drying tert-butyl (±)-erythro-3-hydroxy-l-oxo-3-(pyddin-4yl)-l-(pyrrolidin-l-yl)propan-2-ylcarbamate TTA 08124P (1.30 g, ratio erythro:threo = 96:4 estimated by Ή NMR 300 MHz analysis, 57 % yield) was obtained as a white solid.

OH O (+)-erythro

TTA08124P

MW: 335.40; Yield: 57 %; White Solid; Mp (°C): 170.3

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R_f: 0.45 (EtOAc : MeOH = 85:15).

Ή-NMR (CDC1₃, δ): 1.43 (s, 9H, 3xCH₃), 1.64-1.83 (m, 4H, 2xCH₂), 2.88-2.96 (m, 1H, CH₂N), 3.23-3.29 (m, 1H, CH₂N), 3.34-3.43 (m, 1H, CH₂N), 3.56-3.63 (m, 1H, CH₂N), 4.66 (dd, 1H, J= 8.9 Hz, J= 3.7 Hz, N-CH), 4.91 (dd, 1H, J= 8.4 Hz, J= 3.4 Hz, O-CH), 5.42 (d, 1H, J= 8.8 Hz, OH), 5.75 (d, 1H, J= 8.8 Hz, NH), 7.30 (d, 2H, J= 5.9 Hz, ArH), 8.57 (d, 2H, J= 5.8 Hz, ArH). Ratio erythro'.threo = 96:4.

¹³C-NMR (CDC1₃, δ): 24.0, 25.7, 28.1 (3xC), 45.7, 46.8, 55.2, 74.5, 80.5, 120.9 (2xC), 149.6, 149.8 (2xC), 155.4, 168.8.

MS-ESI m/z (% rel. Int.): 336.1 ([MH]⁺, 10), 280.1 (20), 110.0 (100).

(±)-ezT/Aro-2-Amino-3-hydroxy-3 -(pyridin-4-yl)-1 -(pyrrolidin-1 -yPpropan-1 one dihydrochloride Compound 219.

To a solution of /ez7-butyl (±)-ery7zro-3-hydroxy-l-oxo-3-(pyridin-4-yl)1-(pyrrolidin-l-yl)propan-2-ylcarbamate TTA 08124P (1.15 g, 3.40 mmol) in MeOH (30 mL) in a 250 mL round bottom flask equipped with a magnetic stirrer was added HCI 37 % (3 mL, 35 mmol) at RT. The reaction mixture was stirred at RT for 0.4 h at 45 °C and MeOH was evaporated at 45 °C to give after drying, a white solid TTA 08136 (ratio erythro'.threo = 96:4 estimated by ³H NMR 300 MHz analysis). Amberlite IRA-400 (CT) (10 g) was washed successively with water (2x10 mL), 0.5 N NaOH (3x20 mL), water (2x10 mL) and MeOH (3x10 mL). A solution of TTA 08136 in MeOH (30 mL) was stirred with washed Amberlite IRA-400 for 5 min at RT. After filtration, the MeOH was evaporated and the free base form was purified by column chromatography (SiO₂, CHCl₃:EtOH 95° = 86:14) to give TTA 08136A (445 mg, 55 % yield) as a beige solid (no threo isomer detected by ^!H NMR 300 MHz and HPLC). TTA 08136A (193 mg) was stirred at RT in ethyl acetate (5 mL) with a solution of

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0.1 N HCI in isopropanol (17 mL). Solvents were evaporated at 33 °C, MeOH (0.5 mL) was added and the salt was precipitated by addition of EtOAc (20 mL) to give nearly quantitatively after filtration and drying (±)-ery/Aro-2-ammo-3hydroxy-3 -(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)propan-1 -one dihydrochloride Compound 219 as a white solid (no threo isomer detected by *H NMR 300 MHz and HPLC).

OH O

NH₂

2.HCI (±)-erythro

Compound 219

MW: 308.20; Yield: 55 %; White Solid; Mp (°C): 154.1.

R_z: 0.18 (CHCl₃:EtOH 95° = 86:14, free base).

Ή-NMR (CD₃OD, δ): 1.94-2.04 (m, 4H, 2xCH₂), 3.45-3.56 (m, 2H, CH₂N), 3.66-3.78 (m, 2H, CH₂N), 4.71 (d, 1H, J= 5.2 Hz N-CH), 5.50 (d, 1H, J= 5.1 Hz, O-CH), 8.12 (d, 2H, J= 5.6 Hz, ArH), 8.92 (d, 2H, J= 5.4 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 25.0,27.0, 47.8, 58.4, 70.0, 127.2 (2xC), 143.1 (2xC),

159.8,164.8.

MS-ESI m/z (% rel. Int.): 236.1 ([MH]⁺, 10), 219.1 (55), 110.0 (100).

Preparation of (-)-(27?,37?)-2-amino-3-hydroxy-3-(pyridin-4-yl)-1 -(pyrrolidin-1 yl)propan-l-one dihydrochloride Compound 220 and (+)-(2_>S'.3_iS)-2-amino-3hydroxy-3-(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)propan-1 -one dihydrochloride Compound 221.

Analytical chiral separation:

gL of a 1 mg/mL solution of Compound 219 were injected on

Chiralpak AD: flow-rate - 1 mL/min, temperature = 25 °C, mobile phase:

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152 hexane:ethanol = 8:2, detection UV at 220 nm and by polarimeter, Rt (-) = 16.26 min, Rt (+) = 19.02 min, k(-) = 4.38, k(+) = 5.30, a = 1.21 and resolution Rs= 1.90.

Semi-preparative separation was performed on Chiralpak AS (250x10 mm):

A 40 mg/mL solution of (±)-erjTAro-2-amino-3-hydroxy-3-(pyridin-4-yl)l-(pyrrolidin-l-yl)propan-l-one Compound 219 racemate was prepared and 100 μΙ, of this solution were injected every 4 min on Chiralpak AS, flow-rate = 5 mL/min, mobile phase hexane:ethanol = 1:1, UV detection at 254 nm. 56 successive injections were done. The two main fractions were identified on UV and collected in two different flasks. The solvent was removed in vacuo at 30 °C. The resulting solid was dissolved in 50 mL of CH2CI2 and then filtered on a 0.45 μηι millipore membrane. After evaporation of CH₂C1₂, the solid was dissolved in 50 mL of methanol and then filtered. The salt was regenerated according to the procedure reported for Compound 203 and Compound 204.

The enantiomeric purity of the compounds was checked by analytical injection on Chiralpak AD of the regenerated salts: (-)-(2R,3R)-2-amino-3-hydroxy-3 -(pyridin-4-yl)-1 -(pyrrolidin-1 -yDpropan-1 one dihydrochloride Compound 220.

OH O ¹ NH₂

2.HCI (-)-eryfftro

Compound 220

MW: 308.20; 133 mg obtained; White Solid; Mp (°C): too hygroscopic.

Ry: 0.18 (CHCl₃:EtOH 95° = 86:14, free base).

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Enantiomeric excess = 99.1 % measured by HPLC at 220 nm (Chiralpak AD) a²⁵ _D = - 6.4 (MeOH, c = 1)

Ή-NMR (CD₃OD, δ): 1.94-2.03 (m, 4H, 2xCH₂), 3.45-3.55 (m, 2H, CH₂-N),

3.63-3.76 (m, 2H, CH₂-N), 4.68 (d, 1H, J= 5.1 Hz, N-CH), 5.48 (d, 1H, 5.5

Hz, O-CH), 8.08 (d, 2H, J= 6.4 Hz, ArH), 8.90 (d, 2H, J= 6.6 Hz, ArH).

^,3C-NMR (CD₃OD, δ): 23.4, 25.4,46.2, 56.7, 68.4, 125.7 (2xC), 141.3 (2xC),

158.5, 163.2.

(+)-(25,35)-2-amino-3-hydroxy-3-(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)propan-1 one dihydrochloride Compound 221.

OH O

NH₂ C2.HCI (+)-erythro

Compound 221

MW: 308.20; 140 mg obtained; White Solid; Mp (°C): too hygroscopic.

R_f: 0.18 (CHCl₃:EtOH 95° = 86:14, free base).

Enantiomeric excess = 99.1 % measured by HPLC at 220 nm (Chiralpak AD) a²⁵ _D = + 6.3(MeOH,c=l).

^-NMR (CD3OD, δ): 1.94-2.02 (m, 4H, 2xCH₂), 3.44-3.52 (m, 2H, CH₂-N),

3.64-3.74 (m, 2H, CH₂-N), 4.68 (d, 1H, 5.2 Hz, N-CH), 5.48 (d, 1H, J= 4.3

Hz, O-CH), 8.09 (d, 2H, J= 6.4 Hz, ArH), 8.90 (d, 2H, J= 6.1 Hz, ArH).

¹³C-NMR (CD3OD, δ): 23.4,25.4, 46.2, 56.8, 68.4, 125.7 (2xC), 141.3 (2xC),

158.5,163.2.

Preparation of (±)-/Areo-3-hydroxy-2-(isopropylamino)-3-(pyridin-4-yl)-l(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 223.

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To a stirred solution of (±)-i/ireo-2-amino-3-hydroxy-3-(pyridin-4-yl)-l(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 22 (0.20 g, 0.65 mmol) in 3 mL of MeOH were added triethylamine (180 pl, 1.30 mmol) and acetone (75 pL, 1.00 mmol). The mixture was stirred overnight at RT under nitrogen and then was added AcOH (200 pL, 3.2 mmol) and NaBH₃CN (85 mg, 1.3 mmol). After 5 h at 20 °C, MeOH was evaporated and the residue was partitioned between CH₂C1₂ and 1 N aqueous sodium carbonate. The organic layer was evaporated and the obtained residue was purified by column chromatography (SiO₂, EtOAc:MeOH = 9:1). HCI treatment in MeOH gave (±)/Areo-3-hydroxy-2-(isopropylamino)-3 -(pyridin-4-yl)-1 -(pyrrolidin-1 yl)propan-l-one dihydrochloride Compound 223 (56 mg, 25 % yield) as a white solid.

Compound 223

MW: 350.28; Yield: 25 %; White Solid; Mp (°C): 189.8.

R_f: 0.20 (EtOAc:MeOH= 9:1, free base).

^-NMR (CD₃OD, δ): 1.32 (d, 3H, J= 6.5 Hz, CH₃), 1.40 (d, 3H, J= 6.5 Hz, CH₃), 1.55-1.84 (m, 4H, 2xCH₂), 2.66-2.74 (m, 1H, CH), 3.27-3.54 (m, 4H, CH₂-N), 4.60 (d, 1H, J= 7.9 Hz, N-CH), 5.38 (d, 1H, J= 7.8 Hz, O-CH), 8.24 (d, 2H, J= 6.3 Hz, ArH), 8.98 (d, 2H, J= 6.3 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 18.7, 20.1, 24.7, 26.7, 47.7, 48.5, 52.2, 62.4, 72.5, 127.0 (2xC), 143.2 (2xC), 161.1, 164.1.

MS-ESI m/z (% rel. Int.): 278.1 ([MH]⁺, 25), 179.1 (100).

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HPLC: Method A, detection UV 254 nm. Compound 223 RT = 1.30 min, peak area 99.0 %.

Preparation of (±)-2-amino-3-(hydroxyimino)-3-(pyridin-4-yP-l-(pyrrolidin-lyl)propan-l-one dihydrochloride Compound 224.

(±)-2-fer/-Butyloxycarbonylamino-3-(hydroxyimino)-3-(pyridin-4-vP-l(pyrrolidin-1 -yPpropan-1 -one TTA 08160,

To a stirred solution of /eri-butyl (±)-l,3-dioxo-3-(pyridin-4-yl)-l(pyrrolidin-l-yl)propan-2-ylcarbamate TTA 08120 (0.20 g, 0.60 mmol) in 10 mL of dioxane were added Et₃N (125 μΐ, 0.90 mmol) and hydroxylamine hydrochloride (65 mg, 0.90 mmol). The mixture was stirred 2 h at 110 °C in a sealed tube then dioxane was evaporated. The obtained residue was purified by column chromatography (SiO₂, CH₂Cl₂:MeOH = 9:1 to 97:3) to give (±)-2-teributyloxycarbonylamino-3-(hydroxyimino)-3-(pyridin-4-yl)-1 -(pyrrolidin-1 yl)propan-l-one TTA 08160 (100 mg, 48 % yield) as an oil.

NOH O

TTA 08160

MW: 348.40; Yield: 48 %; Oil.

R_z: 0.30 (CH₂Cl₂:MeOH = 97:3).

^!H-NMR (CDC1₃, δ): 1.39 (s, 9H, 3xCH₃), 1.81-1.98 (m, 4H, 2xCH₂), 3.37-3.62 (m, 4H, 2xCH₂-N), 5.44 (d, 1H, /=8.1 Hz, N-CH), 5.97 (d, 1H, J= 8.1 Hz, NH), 7.37 (d, 2H, /= 5.6 Hz, ArH), 8.65 (d, 2H, J= 6.0 Hz, ArH).

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156 ¹³C-NMR (CDC1₃, δ): 24.0,26.0, 28.2 (3xC), 46.5,46.7, 55.8, 80.3, 123.1 (2xC), 139.7, 149.4 (2xC), 155.1, 166.4.

MS-ESI m/z (% rel. Int.): 349.2 ([MH]⁺, 85), 293.2 (100).

HPLC: Method A, detection UV 254 nm, TTA 08160 RT = 3.90 min, peak area 97.0 %.

(db)-2-Amino-3 -(hydroxyimino)-3-(pyridin-4-yl)-1 -(pyrrolidin-1 -vDpropan-1 one dihydrochloride Compound 224.

(±)-2-/er/-Butyloxycarbonylamino-3-(hydroxyimino)-3-(pyridin-4-yl)-l(pyrrolidin-l-yl)propan-l-one TTA 08160 (100 mg, 0.29 mmol) was dissolved in MeOH (2 mL) and a solution of 1 M HCI in MeOH (1 mL, 3.00 mmol) was added and the mixture was heated for 10 min at 45 °C. MeOH was evaporated and the residue was dried to give crude (±)-2-amino-3-(hydroxyimino)-3(pyridin-4-yl)-l-(pyrrolidin-l-yl)propan-l-one TTA 08164. Amberlite IRA-400 (CT) (1 mL, 1.4 mmol) was washed successively with water (2x10 mL), NaOH 0.5 N (3x20 mL), water (2x10 mL) and MeOH (3x10 mL). A solution of TTA 08164 in MeOH (30 mL) was stirred with washed Amberlite IRA-400 for 5 min at RT. After filtration, the MeOH was evaporated and the free base form was purified by column chromatography (SiO₂, CH₂Cl₂:MeOH = 9:1). HCI Treatment in MeOH gave (±)-2-amino-3-(hydroxyimino)-3-(pyridin-4-yl)-l(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 224 (29 mg, 31 % yield) as a beige solid.

NOHO (±)

Compound 224

MW: 321.20; Yield: 31 %; Beige Solid; Mp (°C): 225.2

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Ry: 0.30 (EtOAc:MeOH= 9:1, free base).

Ή-NMR (CD₃OD, δ): 1.85-2.04 (m, 4H, 2xCH₂), 3.23-3.73 (m, 4H, CH₂-N), 5.58 (d, 1H, J= 4.4 Hz, CH), 8.23 (d, 2H, J= 5.0 Hz, ArH), 9.03 (d, 2H, J= 5.0 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 24.9,26.9, 47.9, 48.0, 55.6, 128.7 (2xC), 143.8 (2xC),

145.5, 148.8, 163.8.

MS-ESI m/z (% rel. Int.): 249.2 ([MH]⁺, 10), 115.0 (100).

HPLC: Method A, detection UV 254 nm, Compound 224 RT = 0.60 min, peak area 99.0 %.

Preparation of (+)-//zreo-2-(4-iodobenzvlamino)-3-hydroxy-3-(pyridin-4-yl)-l(pyrrolidin-l-yl)propan-l-one hydrochloride Compound 225.

To a stirred solution of (±)-^reo-2-amino-3-hydroxy-3-(pyridin-4-yl)-l(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 22 (500 mg, 1.62 mmol) in 7.5 mL of MeOH at RT under nitrogen was added dropwise triethylamine (495 pL, 3.56 mmol) and 4-iodobenzaldehyde (413 mg, 1.73 mmol). The mixture was stirred for 5 h at RT under nitrogen. Acetic acid (463 pL, 8.1 mmol) and NaBH₃CN (356 mg, 5.67 mmol) were added. The mixture was stirred for another 15 h at RT. The mixture was partitioned between EtOAc (750 mL) and an 10 % aqueous solution of potassium carbonate. The organic layer was washed with brine (2 x 20 mL), dried over MgSO₄ and filtered. After evaporation, the crude product was purified by column chromatography (SiO₂, EtOAc:MeOH = 85:15) to give (±)-/Areo-2-(4-iodobenzylamino)-3-hydroxy-3(pyridin-4-yl)-l-(pyrrolidin-l-yl)propan-l-one VIB 01096 as a yellow solid (380 mg, 52 % yield). To a solution of (±)-i/zreo-2-(4-iodobenzylamino)-3hydroxy-3-(pyridin-4-yl)-l-(pyrrolidin-l-yl)propan-l-one VIB 01096 (380 mg,

8.43 mmol) in methanol (10 mL) was added a solution of 0.5 M aqueous

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158 hydrochloric acid (7 mL). After stirring the mixture at RT for 0.5 h the reaction mixture was concentrated and the crude product was coevaporated twice with EtOAc. After trituration with EtOAc, filtration and drying, (±)-/Areo-2-(4iodobenzylamino)-3-hydroxy-3 -(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)propan-1 -one hydrochloride Compound 225 (373 mg, 44 % yield) was obtained as a pale yellow solid.

OH o (±)

Compound 225

MW: 524.37; Yield 44 %; Pale Yellow Solid; Mp (°C): 194.7

Ή-NMR (CD₃OD, δ): 1.42-1.80 (m, 4H, 2xCH₂), 2.32-2.50 (m, 1H, CH₂), 2.96-3.12 (m, 1H, CH₂), 3.12-3.25 (m, 2H, CH₂), 4.27 (q, 2H, J= 13.3 Hz, CH₂), 4.42 (d, 1H, J= 7.8 Hz, HC-N), 5.31 (d, 2H, J= 7.7 Hz, HC-O), 7.29 (d, 2H, J= 8.1 Hz, ArH), 7.80 (dd, 2H, J= 2.9 Hz, J= 8.1 Hz, ArH), 8.15 (d, 2H, J= 5.5 Hz, ArH), 8.91 (d, 2H, J= 5.5 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 24.7, 26.5,47.5,48.4, 51.1, 63.7, 72.3, 96.9, 126.9 (2xC), 131.0, 133.6 (2xC), 139.5 (2xC), 143.2 (2xC), 161.1, 163.8.

MS-ESI m/z (% rel. Int.): 451.9 ([MH]⁺, 100), 363.8 (45), 342.9 (70), 216.9 (75), 148.0 (30).

HPLC: Method A, detection UV 254 nm, Compound 225 RT= 3.83 min, peak area 98.7 %.

Preparation of (±)-/Argo-2-(2-iodobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-l(pyrrolidin-l-yl)-propan-l-one dihydrochloride Compound 226.

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To a stirred solution of (±)-//zreo-2-amino-3-hydroxy-3-pyridin-4-yl-lpyrrolidin-l-yl-propan-l-one Compound 22 (500 mg, 1.62 mmol) in 10 ml methanol was added successively Et₃N (496 pL, 3.57 mmol) and 2-iodobenzaldehyde (414 mg, 1.78 mmol) in 1 ml methanol. The mixture was stirred 6 h at RT under nitrogen and acetic acid (464 pL, 8.11 mmol) and sodium cyanoborohydride (356 mg, 5.57 mmol) were added. The mixture was stirred overnight at 20°C and evaporated to give a residue which was partitioned between CH2Q2 and 1 N aqueous potassium hydroxyde. The organic layer was washed with brine and dried over MgSC>4, filtered and evaporated. The crude product was purified by column chromatography (SiO₂, EtOAc:MeOH = 9:1). After evaporation and drying a white solid (±)-/Areo-2-(2-iodobenzylamino)-3hydroxy-3-(pyridin-4-yl)-l-(pyrrolidin-l-yl)-propan-l-one was obtained (419.6 mg, 57 % yield). The product was dissolved in methanol (10 mL). The solution was stirred at room temperature and a solution of HCI (1 M, 7.4 mL) was added via syringe at RT for 10 min. The mixture was concentrated and triturated with EtOAc. After filtration and drying, (±)-/Azw-2-(2-iodobenzylamino)-3hydroxy-3-(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)-propan-1 -one dihydrochloride Compound 226 was obtained (195 mg, 25 % yield) as a white solid.

OH 0

Compound 226

MW: 524.22; Yield: 25 %; White Solid; Mp (°C): 148.0

R_z: 0.50 (MeOH:EtOAc = 20:80, free base).

Ή-NMR (CD₃OD, δ): 1.33-1.80 (m, 4H, 2xCH₂), 2.12-2.25 (m, 1H, CH₂),

2.75-2.88 (m, 1H, CH₂), 3.14-3.25 (m, 1H, CH₂), 3.25-3.31 (m, 1H, CH₂), 4.14

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160 (d, 1H, J= 8.4 Hz, NH-CH), 4.37-4.52 (m, 2H, NH-CH₂) 5.10 (d, 1H, 7= 8.3 Hz, CH-O), 7.18 (t, 1H, 7= 7.6 Hz, ArH), 7.49 (d, 1H, 7= 7.5 Hz, ArH), 7.61 (d, 1H, 7= 7.7 Hz, ArH), 7.68 (d, 2H, 7= 5.2 Hz, ArH), 7.97 (d, 7= 7.9 Hz, 1H, ArH), 8.66 (d, 7= 4.9 Hz, 2H, ArH).

¹³C-NMR(CD₃OD, δ): 24.7, 26.5, 47.6,47.8, 55.3, 64.3, 72.7, 101.8, 124.4 (2xC), 130.3, 132.0, 132.5, 135.1, 141.7 (2xC), 148.2, 154.0, 164.2. MS-ESI m/z (% rel. Int.): 451.9 ([MH]⁺, 100), 352.9 (55), 342.9 (30). HPLC: Method A, detection UV 254 nm, Compound 226 RT = 3.72 min, peak area 98.95 %.

Preparation of (±)-/Areo-2-(3-iodobenzvlamino)-3-hydroxy-3-(pyridin-4-yl)-l(pyrrolidin-l-yl)-propan-l-one dihydrochloride Compound 227.

To a stirred solution of (±)-/Areo-2-amino-3-hydroxy-3-pyridin-4-yl-lpyrrolidin-l-yl-propan-l-one Compound 22 (500 mg, 1.62 mmol) in 10 ml methanol was added successively Et₃N (496 pL, 3.55 mmol) and 3-iodobenzaldehyde (414 mg, 1.78 mmol) in 1 ml methanol. The mixture was stirred 6 h at RT under nitrogen and acetic acid (464 pL, 8.11 mmol) and sodium cyanoborohydride (356 mg, 5.57 mmol) were added. The mixture was stirred overnight at 20 °C then evaporated and the residue was partitioned between CH₂C1₂ and 1 N aqueous potassium hydroxyde. The organic layer was washed with brine and dried. The organic layer was evaporated. The crude product was purified by column chromatography (SiO₂, CH₂Cl₂:MeOH = 95:5). After evaporation and drying a white solid (±)-/Areo-2-(3-iodobenzylamino)-3hydroxy-3-(pyridin-4-yl)-l-(pyrrolidin-l-yl)-propan-l-one (437 mg, 60 % yield). This product was dissolved in methanol (10 mL). The solution was stirred at RT and a solution of HCI (1 M, 7.7 mL) was added at room temperature for 10 min. The mixture was concentrated and triturated with

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161 diethyl ether. After filtration and drying, (±)-//?reo-2-(3-iodobenzylamino)-3hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin- l-yl)-propan- 1-one dihydrochloride Compound 227 was obtained (205 mg, 26 % yield) as a white solid.

OH o

Compound 227

MW: 524.22; Yield: 26 %; White Solid; Mp (°C): 142.8

Ry: 0.30 (MeOH:CH₂Cl₂ = 5:95, free base).

Ή-NMR (CD₃OD, δ): 1.25-1.80 (m, 4H, 2xCH₂), 2.03-2.12 (m, 1H, CH₂),

2.80-2.92 (m, 1H, CH₂), 3.11-3.25 (m, 2H, CH₂), 4.06-4.30 (m, 3H, CH-N &NCH₂), 5.04 (d, 1H, J= 8.8 Hz, CH-OH), 7.23 (t, 1H, J= 7.8 Hz, ArH), 7.51 (d, 1H, J= 7.5 Hz, ArH), 7.67 (d, 2H, J= 5.8 Hz, ArH), 7.80 (d, 1H, J= 5.2 Hz, ArH), 7.87 (s, 1H, ArH), 8.66 (d, J= 5.2 Hz, 2H, ArH).

¹³C-NMR (CD₃OD, δ): 24.7,26.5, 47.3, 47.7, 50.5, 64.3, 72.8, 95.3, 124.5 (2xC), 130.9, 132.0, 133.9, 140.1,140.5, 148.3, 153.7, 164.2.

MS-ESI m/z (% rel. Int.): 451.9 ([MH]⁺, 100), 352.9 (40), 342.9 (50).

HPLC: Method A, detection UV 254 nm, Compound 227 RT = 3.80 min, peak area 98.7 %.

Preparation of(+)-(2>S',3R)-2-(3-iodobenzylamino)-3-hydroxy-3-(pyridin-4-yl)1-(pyrrolidin-l-yl)-propan-l-one dihydrochloride Compound 228,

To a stirred solution of (+)-(20',3R)-2-amino-3-hydroxy-3-(pyridin-4-yl)1-(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 204 (204 mg, 0.67 mmol) in MeOH (6 mL) was added Et₃N (202 pL, 1.46 mmol) and, via syringe,

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162 a solution of 3-iodobenzaldehyde (169 mg, 0.73 mmol) in methanol (1 mL). The mixture was stirred 5 h at RT under nitrogen. CH₃COOH (190 pL, 3.30 mmol) and sodium cyanoborohydride (146 mg, 2.32 mmol) were added and the reaction mixture was stirred overnight at RT. MeOH was evaporated and the residue was partitioned between CH2CI2 and a solution of 1 N aqueous potassium carbonate. The organic layer was washed with brine, dried over MgSO4, filtered and evaporated. The crude product was purified by column chromatography (SiO₂, CH₂Cl₂:MeOH = 95:5). After evaporation and drying (2S,3R)-2-(3 -iodobenzylamino)-3-hydroxy-3 -(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)propan-l-one (246 mg) was obtained as a white solid. The product was dissolved in methanol (5 mL) and a solution of HCI in MeOH (1 M, 2.5 mL) was added via syringe and the solution was stirred at RT for 0.6 h. The mixture was concentrated and the resulting solid triturated with Et₂O. After filtration and drying (+)-(20',3R)-2-(3-iodobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-l(pyrrolidin-l-yl)-propan-l-one dihydrochloride Compound 228 (280 mg, 80 % yield) was obtained as a white solid.

oh 0

Compound 228

MW: 524.22; Yield: 80 %; White Solid; Mp (°C): 107.4 a²² _D = +101.9 (MeOH, c = 1.02).

R/ 0.30 (MeOH:CH₂Cl₂ = 5:95, free base).

Ή-NMR (CD₃OD, δ): 1.52-1.82 (m, 4H, 2xCH₂), 2.38-2.45 (m, 1H, CH₂),

3.01-3.09 (m, 1H, CH₂), 3.18-3.32 (m, 2H, CH₂), 4.20-4.29 (dd, 2H, J= 28.1

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Hz, J = 13.4 Hz, NH-CH₂), 4.40 (d, 1H, J= 7.9 Hz, N-CH), 5.30 (d, J= 7.9 Hz, 1H, CH-O), 7.25 (t, 1H, J= 5.4 Hz, ArH), 7.54 (d, 1H, J= 7.1 Hz, ArH), 7.81 (d, 1H, J= 7.2 Hz, ArH), 7.89 (d, 1H, J= 1.3 Hz, ArH), 8.13 (d, J= 5.5 Hz, 2H, ArH), 8.90 (d, J= 5.3 Hz, 2H, ArH).

MS-ESI m/z (% rel. Int.): 452.1 ([MH]⁺, 100), 353.0 (65), 343.1 (80). HPLC: Method A, detection UV 254 nm, Compound 228 RT = 3.87 min, peak area 97.0 %.

Preparation of (27?,35)-2-(3-iodobenzylarnino)-3-hydroxy-3-(pyridin-4-yl)-l(pyrrolidin-l-yl)-propan-l-one dihydrochloride Compound 229.

Similar to Compound 228 with (-)-(27?,35)-2-amino-3-hydroxy-3(pyridin-4-yl)-l-(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 203 (77 mg, 0.25 mmol) in 2.3 ml of MeOH, Et₃N (76 pL, 0.79 mmol), 3iodobenzaldehyde (64 mg, 0.275 mmol), CH₃COOH (78.6 pL, 1.37 mmol) and NaBH₃CN (60.5 mg, 0.96 mmol).

(2R,3iS)-2-(3-iodobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-l-(pyrrolidinl-ylj-propan-l-one dihydrochloride Compound 229 (70 mg, 53.5 % yield) was obtained as a pale yellow solid.

OH O

Compound 229

MW: 524.22; Yield: 53.5 %; Pale Yellow Solid; Mp (°C): 179.7 R/ 0.30 (MeOH:CH₂Cl₂ = 5:95, free base).

^-NMR (CD₃OD, δ): idem to Compound 228.

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MS-ESI m/z (% rel. Int.): 452.1 ([MH]⁺, 100), 353.0 (70), 343.1 (60). HPLC: Method A, detection UV 254 nm, Compound 228 RT = 3.78 min, peak area 99.0 %.

Preparation of (±)-//z/^jeo-2-amino-3-hydroxv-1 -morpholino-3-(thiophen-3yl)propan-l-one hydrochloride Compound 230.

£ran,s-(4,5-Dihydro-5-(thiophen-3-yl)oxazol-4-yl)(morpholino)methanone SLA 09052A.

SLA 09052A was prepared in accordance with method D using thiophene-3-carbaldehyde (0.768 mL, 5.35 mmol), KOH (0.273 mg, 4.86 mmol) in methanol (5 mL) and 2-isocyano-l-morpholinoethanone SLA 07118 (0.75 g, 4.86 mmol). The solution was stirred for 2 h at 0 °C. After work-up the residue was purified by column chromatography (florisil, EtOAc). After evaporation and drying, ira«5-(4,5-dihydro-5-(thiophen-3-yl)oxazol-4yl)(morpholino)methanone SLA 09052A (0.327 g, 25 % yield) was obtained as a yellow oil.

(±)-trans

SLA 09052A

MW: 266.32; Yield: 25 %; Yellow Oil.

^-NMR (CDC1₃, δ): 3.40-4.00 (m, 8H, 4xCH₂), 4.67 (dd, 1H, J= 7.3 Hz, J=

2.2 Hz, CH-N), 6.29 (d, 1H, J= Ί.3 Hz, CH-O), 6.97 (d, 1H, 2.2 Hz,

CH=N), 7.01 (dd, 1H, 5.0 Hz, J= 1.3 Hz, CH=C), 7.28-7.40 (m, 2H,

CH=C).

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165 hydrochloride Compound 230.

To a solution of /ra//s-(4,5-dihydro-5-(thiophen-3-yl)oxazol-4yl)(morpholino)methanone SLA 09052A (0.327 g, 1.12 mmol) in MeOH (5 mL) was added HCI 37 % (2 mL). After heating (50 °C) the mixture for 24 h the reaction mixture was concentrated and the crude product was coevaporated twice with EtOAc. After trituration with EtOAc, filtration and drying (±)-i/zreo-

2-amino-3-hydroxy-1 -morpholino-3 -(thiophen-3 -yl)propan-1 -one hydrochloride Compound 230 (276 mg, 77 % yield) was obtained as a pale yellow solid.

Compound 230

MW: 292.78; Yield: 77 %; Pale Yellow Solid; Mp (°C): 209.1 ’H-NMR (CD₃OD, δ): 2.77-2.92 (m, 2H, CH₂), 3.20-3.60 (m, 6H, 3xCH₂), 4.49 (d, 1H, J= 8.7 Hz, CH'N), 4.95 (d, 1H, J= 8.8 Hz, CHO), 7.18 (d, 1H, J= 4.5 Hz, CH=C), 7.44 (d, 1H, J= 1.7 Hz CH=C), 7.44 (dd, 1H, J= 4.5 Hz, 1.7

Hz CH=C).

Preparation of (±)-/Z/reo-2-amino-3-hydroxy-l-(piperidin-l-yl)-3-(thiophen-3yDpropan-l-one hydrochloride Compound 231.

ira7w-(4,5-Dihydro-5-(thiophen-3-yl)oxazol-4-yl)(piperidin-l-yI)methanone

SLA 09052B.

SLA 09052B was prepared in accordance with method D using thiophene-3-carbaldehyde (0.778 mL, 5.43 mmol), KOH (0.273 mg, 4.94

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166 mmol) in methanol (5 mL) and 2-isocyano-l-(piperidin-l-yl)ethanone SLA 07116B (0.75 g, 4.94 mmol). The solution was stirred for 2 h at 0 °C. After work-up (without any further purification) and drying ira«5-(4,5-dihydro-5(thiophen-3-yl)oxazol-4-yl)(piperidin-l-yl)methanone SLA 09052B was obtained as a yellow oil (1.29 g, 99 % yield).

(+)-trans

SLA 09052B

MW: 264.34; Yield: 99 %; Yellow Oil.

*H-NMR (CDCls, δ): 1.47-1.75 (m, 6H, 3xCH₂), 3.42-3.82 (m, 4H, 2xCH₂),

4.72 (dd, 1H, J= 7.2 Hz, 2.2 Hz, CH-N), 6.28 (d, 1H, J= 7.2 Hz, CH-O), 6.96 (d, 1H, J= 2.2 Hz, CH=N), 7.01 (dd, 1H, J= 5.0 Hz, J= 1.2 Hz, CH=C),

7.30-7.35 (m, 2H, CH=C).

¹³C-NMR (CDCI3, δ): 24.5, 25.5, 26.5, 43.7, 46.8, 73.4, 78.0, 122.6,125.1,

127.2, 140.5, 155.0, 166.3.

(±)-^reo-2-Amino-3-hydroxy-1 -(piperidin-1 - yl)-3 -(thiophen-3 -yDpropan-1 -one hydrochloride Compound 231.

To a solution of irans-(4,5-dihydro-5-(thiophen-3-yl)oxazol-4yl)(piperidin-l-yl)methanone SLA 09052B (1.29 g, 4.88 mmol) in methanol (5 mL) was added hydrochloric acid 37 % (5 mL). After heating (50 °C) the mixture for 24 h the reaction mixture was concentrated and the crude product was coevaporated twice with EtOAc. After trituration with EtOAc, filtration and drying (±)-rizreo-2-amino-3-hydroxy- l-morpholino-3-(thiophen-3-yl)propan-1WO 2006/081273

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167 one hydrochloride Compound 231 was obtained as a pale yellow solid (1.07 g, % yield).

(±)

Compound 231

MW: 290.81; Yield: 75 %; Pale Yellow Solid; Mp (°C): 210.6 Ή-NMR (CD₃OD, δ): 0.82-0.97 (m, 1H, 0.5xCH₂), 1.30-1.70 (m, 5H, 2.5xCH₂), 2.75-2.91 (m, 1H, CH₂), 3.12-3.55 (m, 3H, 1.5xCH₂), 4.51 (d, 1H, J = 8.4 Hz, CH’N), 4.94 (d, 1H, J= 8.5 Hz, CHO), 7.16 (d, 1H, J= 4.9 Hz, CH=C), 7.44 (d, 1H, J= 2.5, Hz CH=C), 7.44 (dd, 1H, J= 4.5 Hz, J= 3.2 Hz CH=C).

Preparation of (±)-riireo-2-amino-3-hydroxy-1 -morpholino-3-(pyridin-3vl)propan-l-one dihydrochloride Compound 232.

frans-(4,5-Dihydro-5-(pyridin-3-yl)oxazol-4-yl)(morpholino)methanone SLA 09050A.

SLA 09050A was prepared in accordance with method D using pyridine-

3-carbaldehyde (0.65 mL, 4.86 mmol), KOH (0.273 mg, 4.86 mmol) in methanol (10 mL) and 2-isocyano-l-morpholinoethanone SLA 07118 (0.75 g, 4.86 mmol). The solution was stirred for 20 h at 0 °C. After work-up (without any further purification), evaporation and drying trans-(A,5-dihydro-5-(pyridin-

3-yl)oxazol-4-yl)(morpholino)methanone SLA 09050A was obtained as a yellow solid (0.92 g, 72.5 % yield).

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N ⁰ </° (±)-trans

SLA 09050A

MW: 261.28; Yield: 72.5 %; Yellow Solid.

^-NMR (CDCI3, δ): 3.42-4.00 (m, 8H, 4xCH₂), 4.63 (dd, 1H, J= 7.7 Hz, J=

2.3 Hz, CH-N), 6.29 (d, 1H, J= 7.7 Hz, CH-O), 7.02 (d, 1H, J= 2.2 Hz, CH=N), 7.33 (m, 1H, ArH), 7.60-7.66 (m, 1H, ArH), 8.57-8.62 (m, 2H, ArH). ¹³C-NMR (CDCI3, δ): 43.0,46.3, 66.7, 66.8, 74.6, 79.2, 123.5, 133.4, 135.1,

147.5, 150.0, 155.0, 166.3.

MS-ESI m/z (% rel. Int.): 262.1 ([MH]⁺, 55), 108.0 (100).

(±)-/Areo-2-Amino-3-hydroxy-1 -morpholino-3 -(pyridin-3 -yDpropan-1 -one dihydrochloride Compound 232.

To a solution of trans-(4,5-dihydro-5-(pyridin-3-yl)oxazol-4yl)(morpholino)methanone SLA 09050A (0.911 g, 3.48 mmol) in methanol (10 mL) was added hydrochloric acid 37 % (5 mL). After heating (50°C) the mixture for 2.25 h the reaction mixture was concentrated and the crude product was coevaporated twice with EtOAc and the crude product was dissolved in a 1 N solution of K₂CO₃ which was extracted with a mixture CH₂Cl₂:iPrOH = 9:1 (6 x 100 mL). The combined organic layer was dried over MgSO₄, filtered and evaporated to led to a crude product which was purified by column chromatography (SiO₂, EtOAC:MeOH = 70:30). After evaporation the product was dissolved in a solution of HCI in MeOH (0.5M, 29 mL) and stirred at RT for 1.5 h. The product was co-evaporated twice with EtOAc. After trituration with EtOAc, filtration and drying, (±)-iAreo-2-amino-3-hydroxy-1-morpholinoWO 2006/081273

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3-(pyridin-3-yl)propan- 1-one dihydrochloride Compound 232 (270 mg, 24 % yield) was obtained as a yellow solid.

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169

Compound 232

MW: 324.2; Yield: 24 %; Yellow Solid.

^-NMR (CD₃OD, δ): 3.20-3.78 (m, 8H, 4xCH₂), 4.87 (d, 1H, J= 5.5 Hz, CH’ N), 5.40 (d, 1H, J= 5.4 Hz, CHO), 8.20 (m, 1H, ArH), 8.77 (d, 1H, 8.3 Hz,

ArH), 8.93 (d, 1H, J= 5.5 Hz, ArH), 9.03 (s, 1H, ArH).

Preparation of (±)-^reo-2-amino-3-hydroxy-1 -piperidin-3-(pyridin-3yl)propan-l-one dihydrochloride Compound 233.

imn5^,-(4,5-Dihydro-5-(pvridin-3-yl)oxazol-4-yl)(piperidin)methanone SLA 09050B.

SLA 09050B was prepared in accordance with method D using pyridine-

3-carbaldehyde (0.512 mL, 5.42 mmol), KOH (0.277 mg, 4.93 mmol) in methanol (10 mL) and 2-isocyano-l-piperidinethanone SLA 07116B (0.75 g, 4.93 mmol). The solution was stirred for 20 h at 0 °C. After work-up (without any further purification), evaporation and drying, irans-(4,5-dihydro-5-(pyridin-

3-yl)oxazol-4-yl)(piperidin)methanone SLA 09050B (0.917 g, 72 % yield) was obtained as a yellow solid.

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SLA 09050B

MW: 259.30; Yield: 72 %; Yellow Solid.

’H-NMR (CDC1₃, δ): 1.59-1.73 (m, 6H, 3xCH₂), 3.44-3.83 (m, 4H, 2xCH₂),

4.68 (dd, 1H, J= 7.6 Hz, J= 2.3 Hz, CH-N), 6.30 (d, 1H, J= 7.6 Hz, CH-O), 7.02 (d, 1H, J= 2.2 Hz, CH=N), 7.32 (m, 1H, ArH), 7.60-7.66 (m, 1H, ArH), 8.57-8.62 (m, 2H, ArH).

¹³C-NMR (CDC1₃, δ): 24.4, 25.5, 26.4,43.8, 46.9, 74.6, 79.5, 123.6,133.5,

135.4, 147.5, 149.9, 154.8, 165.9.

(±)-^reo-2-Amino-3-hydroxv-1 -piperidin-3-(pyridin-3-yl)propan-1 -one dihydrochloride Compound 233.

To a solution of iran5-(4,5-dihydro-5-(pyridin-3-yl)oxazol-4yl)(piperidin)methanone SLA 09050B (0.917 g, 3.54 mmol) in methanol (10 mL) was added hydrochloric acid 37 % (5 mL). After heating at 50 °C the mixture for 2.25 h the reaction mixture was concentrated and the crude product was coevaporated twice with EtOAc. After trituration with EtOAc, filtration and drying, the crude product was dissolved in a 1 N solution of K₂CO₃ and the product was extracted CH₂Cl₂:iPrOH = 9:1 (6 x 100 mL). The crude product was purified by column chromatography (EtOAc:MeOH = 7:3) (±)-threo-2amino-3-hydroxy-l-piperidin-3-(pyridin-3-yl)propan-l-one was obtained pale yellow solid (223.5 mg). The product was dissolved in a solution of HCI in MeOH (0.5 M, 18 mL) and stirred at RT for 1.5 h. After trituration with EtOAc, filtration and drying, (±)-/Areo-2-amino-3-hydroxy-l-piperidin-3-(pyridin-3yl)propan-l-one dihydrochloride Compound 233 (208 mg, 18 % yield) was obtained as a yellow solid.

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OH O

HCI (±)

Compound 233

MW: 322.2; Yield: 18 %; Yellow Solid.

Ή-NMR (CD₃OD, δ): 1.15-1.80 (m, 6H, 4xCH₂), 3.10-3.80 (m, 4H, 2xCH₂), 4.88 (d, 1H, J= 5.6 Hz, CH'N), 5.33 (d, 1H, J= 5.6 Hz, CHO), 8.19 (t, 1H, J=

7.1 Hz, ArH), 8.74 (d, 1H, J= 7.9 Hz, ArH), 8.93 (d, 1H, J= 5.6 Hz, ArH), 9.01 (s, 1H, ArH).

Preparation of (2R,3S)-2-((R)-2-hydroxy-l-phenylethylamino)-3-hydroxv-3(pyridin-4-yl)-l-(pyrrolidin-1-vPpropan-1-one dihydrochloride Compound 234.

(R)-5-Phenylmorpholin-2-one EBE 06134.

To a solution of phenylbromoacetate (18.58 g, 86 mmol) in CH₃CN under nitrogen was added a solution of (R)-phenylglycinol (10.17 g, 74 mmol) and diisopropylethylamine (34 mL, 195 mmol) in CH₃CN. The volatiles were removed under reduced pressure keeping the bath temperature below 25 °C to obtain an oil that was treated with EtOAc (120 mL) and stirred for 15 min. The resulting white precipitate was removed by filtration. The filtrate was concentrated under reduced pressure and the desired product was isolated using column chromatography (SiO₂) with a step gradient of 10 % to 50 % [v/v] EtOAc in cyclohexane to give after evaporation (7?)-5-phenylmorpholin-2-one EBE 06134 (3.17 g, 24 % yield) as a white solid.

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MW: 177.2; Yield: 24 %; White Solid; Mp (°C): 50.3

Ry·: 0.30 (EtOAc:cyclohexane = 50:50).

Ή-NMR (CDCls, δ): 1.99 (s, 1H, NH), 3.89 (q, 2H, J= 17.8 Hz, N-CH₂), 4.18 (dd, 1H, J= 3.7 Hz, J= 10.3 Hz, O-CH), 4.29 (t, 1H, J= 10.5 Hz, N-CH), 4.40 (dd, 1H, J= 3.7 Hz, J= 10.5 Hz, O-CH), 7.30-7.45 (m, 5H, ArH).

¹³C-NMR (CDC1₃, δ): 46.8, 54.8, 72.8, 125.3 (2xC), 127.0, 127.3 (2xC), 135.9, 166.0.

[a]²² _D = + 30.3 ⁰ (c = 1.00, MeOH).

(l£3R,5R,8aR)-Tetrahydro-5-phenyl-l,3-di(pyridin-4-yl)oxazolor4,3cirL4~]oxazin-8(3ZZ)-one EBE 06136.

A solution of (R)-5-phenylmorpholinon-2-one EBE 06134 (3.0 g, 16.9 mmol) and pyridine-4-carboxaldehyde (5.43 g, 50.7 mmol) in toluene (75 mL) was refluxed in a soxhlet extractor filled with molecular sieves 4A (25 g) for 16 hours. All the volatiles were evaporated and the desired product was purified by column chromatography (SiO₂) using a gradient of 80 % to 100 % [v/v] EtOAc in cyclohexane to give after evaporation (l₁S',3E,5R,8aR)-tetrahydro-5-phenyll,3-di(pyridin-4-yl)oxazolo[4,3-c][l,4]oxazin-8(3Z7)-one EBE 06136 (1.7 g, 46 % yield) as a pale yellow solid.

EBE 06136

MW: 373.4; Yield: 46 %; Pale Yellow Solid; Mp (°C): 155.6

R_y: 0.20 (EtOAc).

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Ή-NMR (CDC1_3j δ): 4.10-4.17 (m, 1H, N-CH), 4.25 (dd, 1H, J = 3.3 Hz, NCH), 4.36-4.54 (m, 2H, O-CH), 5.38 (d, 1H, O-CH, J= 8.2 Hz), 5.53 (s, 1H, NCH), 7.20-7.35 (m, 8H, ArH), 7.40-7.50 (m, 1H, ArH), 8.51 (dd, 2H, <7= 1.4 Hz, J= 4.5 Hz), 8.57 (dd, 2H, J = 1.4 Hz, J = 4.5 Hz, ArH).

((27?,45,57?)-3-((7?)-2-Hydroxy-l-phenylethyl)-2,5-di(pyridin-4-yl)oxazolidin-4yl)(pyrrolidin-l-yl)methanone EBE 06138.

To a solution of (lS,3R,5R,8aR)-tetrahydro-5-phenyl-l,3-di(pyridin-4yl)oxazolo[4,3-c][l,4]oxazin-8(377)-one EBE 06136(1.7 g, 4.55 mmol) in CH2CI2 was added pyrrolidine (1.90 mL, 22.8 mmol) and the solution was stirred under nitrogen at 25 °C for 16 h. All the volatiles were evaporated and the resulting product was isolated using column chromatography (SiO₂) with a gradient of 0-20 % [v/v] MeOH in EtOAc to give after evaporation ((27?,46*,57?)3-((7?)-2-hydroxy-l-phenylethyl)-2,5-di(pyridin-4-yl)oxazolidin-4yl)(pyrrolidin-l-yl)methanone EBE 06138 (0.665 g, 33 % yield) as a white solid.

EBE 06138

MW: 444.5; Yield: 33 %; White Solid; Mp (°C): 63.6.

R_z: 0.25 (MeOH:EtOAc = 20:80).

Ή-NMR (CDC1₃, δ): 1.70-1.90 (m, 4H, 2xCH₂), 2.25 (bs, 1H, OH), 2.75-2.85 (m, 1H, CH-N), 2.95-3.05 (m, 1H, N-CH), 3.50-3.60 (m, 2H, N-CH₂), 3.80-4.15 (m, 4H, 2xCH +CH₂-O), 5.10 (d, 1H, J= 4.7 Hz, CH), 6.32 (s, 1H, CH), 7.18WO 2006/081273

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7.32 (m, 7H, ArH), 7.45 (d, 2H, J= 5.9 Hz, ArH), 8.54 (d, 2H, J= 6.0 Hz, ArH), 8.64 (d, 2H, J= 6.0 Hz).

¹³C-NMR (CDC1₃, δ): 23.9, 26.0, 46.3, 46.6, 60.3, 63.8, 65.2, 94.1, 121.1 (2xC), 123.0 (2xC), 128.0, 128.3 (2xC), 128.5 (2xC), 137.8, 147.8. MS-ESI m/z (% rel. Int.): 445.1 ([MH]⁺, 20).

HPLC: Method A, detection at 254 nm, EBE 06138 RT = 3.50 min, peak area 99 %.

[a]²²D = -16.0 ⁰ (c = 1.00, CHC13).

(2R,35^f)-2-((R)-2-Hvdroxv-l-nhenylethylamino)-3-hydroxv-3-(r)yridin-4-vB-l(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 234.

To a solution of ((2R,4S,5R)-3-((R)-2-hydroxy-l-phenylethyl)-2,5di(pyridin-4-yl)oxazolidin-4-yl)(pyrrolidin-l-yl)methanone EBE 06138 (600 mg, 1.34 mmol) in MeOH (6 mL) was added a solution of 1 N HCI (6 mL) and the reaction mixture was stirred at 60 °C for 2 h, evaporated to dryness to give a white solid (745 mg). This crude product was dissolved in CH2CI2 (10 mL) and Na₂CO₃ (sat. sol.). The organic layer was separated was separated and the aqueous layer was washed with CH₂C1₂ (5x2 mL). The combined organic layer were dried over Na₂SO4, filtered over cotton mixed with silica gel (600 mg), evaporated and loaded on a silica gel column of 25 g. The desired product was eluted using a gradient of MeOH 0 % to 20 % in EtOAc to give after evaporation (2R,36)-2-((R)-2-hydroxy-l-phenylethylamino)-3-hydroxy-3(pyridin-4-yl)-l-(pyrrolidin-l-yl)propan-l-one (320 mg, 67 % yield) as a white solid.

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OH O

Compound 234

MW = 355.43; Yield: 67 %; White Solid; Mp (°C) = 76.4

R_f: 0.3 (MeOH:EtOAc = 20:80).

^NMRCCDCls, δ): 1.18-1.30 (m, 2H, CH₂), 1.30-1.48 (m, 2H, CH₂), 1.80-

1.90 (m, 1H, CH₂), 2.23-2.33 (m, 1H, N-CH₂), 2.85-2.95 (m, 2H, N-CH₂), 3.003.12 (m, 2H, N-CH + N-CH), 3.72 -3.85 (m, 3H, O-CH₂ + NH), 4.58 (d, 1H, J=

8.4 Hz, O-CH), 7.18-7.35 (m, 7H, ArH), 8.51 (d, 2H, J= 6.0 Hz, ArH).

[a]²²D = - 68.8 ⁰ (c = 1.00, CHC13).

To a solution of (25,35)-2-((5)-2-hydroxy-l-phenylethylamino)-3hydroxy-3-(pyridin-4-yl)-l-(pyrrolidin-l-yl)propan-l-one in MeOH (100 mg), in MeOH (1 mL) was added a solution HCI (IN, 1.1 mL) at 0 °C and the solution was stirred for 10 min to give after evaporation (27?,35)-2-((7?)-2hydroxy-1 -phenylethylamino)-3 -hydroxy-3 -(pyridin-4-yl)-1 -(pyrrolidin-1 yl)propan-l-one dihydrochloride Compound 234 (120 mg, 99 % yield) as a white solid.

OH o

HCI

Compound 234

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MW: 430.4; Global Yield: 66 %; White Solid; Mp (°C): 38.6. Ή-NMR (CD₃OD, δ): 1.38-1.58 (m, 4H, CH₂), 2.12-2.22 (m, 1H, N-CH₂),

2.80-2.90 (m, 1H, N-CH₂), 3.00-3.10 (m, 2H, N-CH₂), 3.91 (dd, 1H, J= 4.3 Hz, 7= 11.3 Hz, CH₂-O), 4.02-4.12 (m, 1H, CH₂-O), 4.55-4.65 (m, 2H, N-CH), 5.27 (d, 1H, J= 8.9 Hz, 7.35-7.45 (m, 3H, ArH), 7.45-7.58 (m, 2H, ArH). ¹³C-NMR (CD₃OD, δ): 24.5, 26.3, 47.2,48.0, 63.5, 64.4, 67.3, 72.6, 126.7,

130.2,131.5,132.3, 143.3, 160.7, 163.8.

Preparation of 7V-((±)-Z/zreo- l-hydroxy-3-oxo- l-(pyridin-4-yl)-3-(pyrrolidin-1 yl)propan-2-yl)acetamide hydrochloride Compound 235.

To a suspension of (±)-//?reo-2-amino-3-hydroxy-3-(pyridin-4-yl)-l(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 22 (300 mg, 0.89 mmol) in CH₂C1₂ (6 mL) was added Et₃N (370 pL, 2.67 mmol) and the mixture was stirred for 10 min, cooled at 4 °C and a solution of acetic anhydride (65 mL, 0.89 mmol) in CH₂C1₂ was added dropwise for 10 min. The reaction mixture was brought to room temperature, stirred for 16 h and washed with water (3x4 mL), NaOH (0.5 N) (3x4 mL) evaporated to give an oily residue that was purified using column chromatography (SiO₂) with a gradient of 0 % to 20 % MeOH in EtOAc. The product in MeOH at 4 °C was treated with a solution of 1 N HCI (4 mL) and all the volatiles were evaporated. Product was precipitated using a mixture of methanol in EtOAc to obtain after evaporation N~((±ythreo-1 -hydroxy-3 -oxo-1 -(pyridin-4-yl)-3-(pyrrolidin-1 -yl)propan-2yl)acetamide hydrochloride Compound 235 (209 mg, 52 % yield).

oh o (±)

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Compound 235

MW: 313.78; Yield: 52 %; White Solid; Mp (°C): 181.3 Ry·: 0.20 (MeOH:EtOAc = 20:80, free base).

Ή-NMR (CD₃OD, δ): 1.82-2.05 (m, 7H, 2*CH₂ & CH₃), 3.35-3.45 (m, 2H, CH₂), 3.50-3.65 (m, 2H, CH₂), 5.10 (d, 1H, J= 3.8 Hz, N-CH), 5.39 (d, 1H,

3.9 Hz, O-CH), 8.16 (d, 2H, J= 6.2 Hz, ArH), 8.81 (d, 2H, J= 6.7 Hz, ArH). ¹³C-NMR (CD₃OD, δ): 22.1, 25.0,27.0,47.5, 48.2, 57.0, 72.0, 126.6 (2xC), 142.0 (2C), 142.0 (2xC), 164.4,169.0,173.0.

Preparation of (R)-2-amino-3-(pyridin-4-yP-1 -(pyrrolidin-1 -yllpropan-1 -one dihydrochloride Compound 236.

terf-Butyl (R)-1 -oxo-3-(pyridin-4-yP-1 -(pyrrolidin-1 -yl)propan-2-ylcarbamate EBE 06172.

W-Boc-(S)-3-(pyridin-4-yl)alanine (0.500 g, 1.88 mmol) was dissolved in CH₂C1₂ (15 mL) and DIEA (361 μι, 2.07 mmol) was added. The mixture was cooled to 0 °C and isobutyl chloroformate (270 pL, 2.07 mmol) was added. The mixture was stirred for 10 min and pyrrolidine (267 mg, 3.76 mmol) in CH₂C1₂(5 mL) was added. This mixture was stirred for 15 min at 4 °C, 12 h at 25 °C, washed successively with NaH₂PO4, saturated sodium hydrogen carbonate, water and brine. The organic layer was dried over magnesium sulfate and evaporated to dryness. The residue obtained was purify by column chromatography (SiO₂) using a gradient of MeOH 0-10% [v/v] in EtOAc to give /eri-butyl (7?)-1 -oxo-3 -(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)propan-2ylcarbamate EBE 06172 (146 mg, 24 % yield) as a white solid.

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HN '—' )=0

EBE 06172

MW: 319.4; Yield: 24 %; White Solid; Mp (°C): 52.5.

Ry: 0.30 (MeOH:EtOAc = 20:80).

Ή-NMR (CDC1₃, δ): 1.40 (s, 9H, (CH₃)₃), 1.65-1.90 (m, 4H, CH₂), 2.80-2.90 (m, 1H, CH), 2.90-3.00 (m, 2H, CH₂), 3.28-3.38 (m, 1H, N-CH), 3.40-3.50 (m, 2H, N-CH₂), 4.60-4.70 (m, 1H, N-CH), 5.38-5.48 (m, 1H, NH), 7.16 (d, 2H, J=

4.5 Hz, ArH), 8.50 (d, 2H, 7= 4.5 Hz, ArH).

MS-ESI m/z (% rel. Int.): 320.2 ([MH]⁺, 20).

HPLC: Method A, detection at 254 nm, EBE 06172 RT = 3.82 min, peak area 85 %.

(R)-2-Amino-3-(pyridin-4-yP-1 -(pyrrolidin-1 -yPpropan-1 -one dihydrochloride Compound 236.

To a solution of TFA (2 mL) in CH₂C1₂ (8 mL) was added (7?)-l-oxo-3(pyridin-4-yl)-l-(pyrrolidin-l-yl)propan-2-ylcarbamate EBE 06172 (146 mg, 0.654 mmol) and the mixture was stirred for 2 h at 25 °C. The volatiles were evaporated and the product was treated with a suspension on amberlite-400 (OH form, 2 g) in MeOH. The suspension was filtered and washed with MeOH (3x5 mL). The combined methanol fractions were evaporated under reduced pressure and the desired product was isolated using column chromatography (SiO₂) with a mixture of EtOAc:MeOH:NH4OH = 70:30:4 to give an oily residue that was treated with a solution of 0.1 N HCI in iPrOH for 10 min. Evaporation of the volatile afforded (R)-2-amino-3-(pyridin-4-yl)-l-(pyrrolidinWO 2006/081273

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Compound 236

MW: 255.75; Yield: 47 %; Pale Yellow Solid; Mp (°C): 127.5 R_z: 0.30 (EtOAc:MeOH:NH₄OH = 70:30:4, free base).

Ή-NMR (CD₃OD, δ): 1.80-2.05 (m, 4H, CH₂), 3.28-3.80 (m, 6H, CH₂, N-CH₂),

3.70 (t, 1H, J= 6.7 Hz, CH-N), 8.10 (d, 2H, J= 5.9 Hz, ArH), 8.88 (d, 2H, J=

5.6 Hz, ArH).

¹³C-NMR(CD₃OD, δ): 23.4,25.4, 35.6,46.1, 46.5, 51.1, 128.5 (2*C), 146.3 (2*C), 156.1, 165.0.

Preparation of /er/-butyl 5-((+)-//?reo-l-hydroxy-3-oxo-l-(pyridin-4-yl)-3(pyrrolidin-1 -yl)propan-2-ylcarbamoyl)pentylcarbamate Compound 237.

To a solution of WBoc-aminohexanoic acid (342 mg, 1.48 mmol) in THF (10 mL) was added Mmethylmorpholine (163 pL, 1.48 mmol). The solution was stirred for 5 min, cooled at -15 °C and treated dropwise with isobutyl chloroformate (211 pL, 1.48 mmol). This solution was added via a stainless steal cannula to a solution of (±)-i/zreo-2-amino-3-hydroxy-3-(pyridin-4-yl)-l(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 22 (500 mg, 1.48 mmol) and Mmethyl-morpholine (489 mg, 1.47 mmol) in THF (10 mL) at 15°C. The reaction mixture was kept for 0.5 h at -15 °C followed by 2 h at 25 °C with continuous stirring. After evaporation of the solvent, the residue was partitioned between EtOAc and H₂O, washed with NaH₂PO₄, saturated aqueous

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NaHCO₃, dried over sodium sulfate and purified by column chromatography (SiO₂) with a gradient of 0 % to 10 % [v/v] MeOH in EtOAc to give tert-butyl

5-((±)-threo-1 -hydroxy-3-oxo-1 -(pyridin-4-yl)-3-(pyrrolidin-1 -yl)propan-2ylcarbamoyl)pentylcarbamate Compound 237 (455 mg, 69 % yield) as a white solid.

OH o

Compound 237

MW: 448.6; Yield: 69 %; White Solid.

Ry: 0.20 (EtOAc:MeOH = 90: 10).

Ή-NMR (CD₃OD, δ): 1.05-1.15 (m, 2H, CH₂), 1.35-1.55 (m, 13H, 2*CH₂ + C(CH₃)₃), 1.75-1.95 (m, 4H, 2*CH₂), 2.00-2.20 (m, 2H, O=CCH₂), 3.05 (q, 2H, J= 6.7 Hz, N-CH₂), 3.20-3.35 (m, 1H, N-CH), 3.38-3.50 (m, 2H, N-CH₂), 3.653.75 (m, 1H, N-CH), 4.72 (bs, 1H, NH), 4.98 (dd, 1H, J= 8.8 Hz, J= 3.6 Hz), 5.08 (d, 1H, J= 3.3 Hz, OCH), 5.23 (bs, 1H, OH), 6.50 (d, 1H, J= 8.7 Hz, NH),

7.35 (d, 2H, J= 6.0 Hz, ArH), 8.58 (d, 2H, J= 4.6 Hz, J= 1.4 Hz, ArH).

MS-ESI m/z (% rel. Int.): 449.2 ([MH]⁺, 30), 349.2 (100).

HPLC: Method A, detection at 254 nm, Compound 237 RT = 4.03 min, peak area 99.9 %.

Preparation of 6-amino-7V-((±)-/Areo-l-hydroxy-3-oxo-l-(pyridin-4-yl)-3(pyrrolidin-l-yl)propan-2-yl)hexanamide Compound 238.

To a solution of tert-butyl 5-((±)-/Areo-l-hydroxy-3-oxo-l-(pyridin-4-yl)3-(pyrrolidin-l-yl)propan-2-ylcarbamoyl)pentylcarbamate Compound 237 (81

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181 mg, 0.181 mmol) in CH₂C1₂ (8 mL) was added TFA (2 mL) at 0 °C and stirred for 2 h at 0 °C. All the volatiles were evaporated to give a residue that was treated with a suspension of Amberlite-400 (OH') in MeOH. After filtration, the filtrate was evaporated and the product was isolated by column chromatography (SiO₂) with CH₂Cl₂:MeOH:NH₄OH = 10:5:0.4 to afford 6-amino-W-((±)-/Areo1 -hydroxy-3-oxo-1 -(pyridin-4--yl)-3-(pyrrolidin-1 -yl)propan-2-yl)hexanamide Compound 238 (40 mg, 64 % yield) as a white solid.

Compound 238

MW: 448.6; Yield: 64 %; White Solid; Mp (°C): 134.4

R/ 0.30 (CH₂Cl₂:MeOH:NH₄OH = 10:5:0.4).

‘HNMR (CDCI3, δ): 1.12-1.30 (m, 2H, CH₂), 1.30-1.50(m, 2H, CH₂), 1.50-1.65 (m, 2H, CH₂), 1.65-1.95 (m, 4H, CH₂), 2.10-2.30 (m, 2H, CH₂), 2.55-2.70 (t, 2H, J= 6.9 Hz, CH₂), 3.10 -3.20 (m, 2H, CH₂), 3.28-3.50 (m, 2H, CH₂), 3.60-

3.70 (m, 1H, CH), 4.95 (dd, 1H, 5.1 Hz, J = 8.4Hz, O-CH), 5.02 (d, 1H, J=

5.0 Hz, OH), 7.11 Hz (d, J= 8.48 Hz, 1H, ArH), 7.35 (dd, 2H, J= 4.4 Hz, J =

1.5 Hz, ArH), 8.55 (dd, J= 1.5 Hz, J= 4.6 Hz, 2H, ArH).

¹³C NMR (CDCI3, δ): 24.0, 25.1, 25.8, 25.9, 32.5, 35.8, 41.7,46.0, 46.9, 55.6,

72.6, 121.3 (2xC), 149.2, 149.5 (2*C), 168.9, 173.7.

Preparation of (±)-ery//zro-2-amino-3-hydroxy-3-(pyridin-4-yl)-l-(pyrrolidin-lyl)butan-l-one dihydrochloride Compound 239.

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182 (+)-cz5^,-(5-Methvl-5-pyridin-4-yl-4,5-dihydro-oxazol-4-yl)-pyrrolidin-l-ylmethanone EBE 06180.

To a stirred solution of KOH (223 mg, 39.7 mmol) in MeOH was added

4-acetylpyridine (478 mg, 39.7 mmol) and 2-isocyano-l-(pyrrolidin-lyl)ethanone BLE 04098 (500 mg, 3.2 mmol). The reaction mixture was stirred for 3 h at 0 °C and then concentrated. The residue was partitioned between EtOAc and H₂O. The organic layer was washed with brine, dried over MgSO₄, filtered and evaporated to give a yellow oil that was purified by column chromatography (SiO₂) with 20 % [v/v] MeOH in EtOAc to give cis- and trans(±)-(5-methyl-5-pyridin-4-yl-4,5-dihydro-oxazol-4-yl)-pyrrolidin-l-ylmethanone. The mixture was further purified by column chromatography (SiO₂) using a gradient of 2 % to 5 % [v/v] MeOH [v/v] in CH₂C1₂ to obtain the pure cz0^,-(±)-(5-methyl-5-pyridin-4-yl-4,5-dihydro-oxazol-4-yl)-pyrrolidin-l-ylmethanone EBE 06180 (122 mg, 51 % yield) as white solid.

EBE 06180

MW: 259.3; Yield: 51 %; White Solid; Mp (°C): 140.9

R_z: 0.30 (EtOAc:MeOH = 80:20).

Ή-NMR (CDC1₃, δ): 1.45-1.75 (m, 4H, 2xCH₂), 1.81 (s, 3H, CH₃), 2.75-2.90 (m, 1H, N-CH₂), 3.10-3.22 (m, 1H, N-CH₂), 3.30-3.40 (t, 2H, J= 6.7 Hz, NCH₂), 4.83 (d, 1H, J= 1.7 Hz, NCH), 7.22 (d, 1H, J= 1.4 Hz, N=CH), 7.27 (d, 2H, J = 6.0 Hz, Ar), 8.57 (d, 2H, J= 6.1 Hz, ArH).

¹³C-NMR (CDC1₃, δ): 23.6, 26.0,27.9, 46.0, 46.5, 77.8, 87.2, 120.2, 148.9,

149.6 (2xC), 155.6 (2xC), 165.4.

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183 (±)-ery/Aro-2-Amino-3-hydroxy-3-(pyridin-4-yl)-1 -(pyrrolidin-1 -yDbutan-1 -one dihydrochloride Compound 239.

To a solution czs-(±)-(5-methyl-5-pyridin-4-yl-4,5-dihydro-oxazol-4-yl)pyrrolidin-l-yl-methanone EBE 06180 (50 mg, 0.19 mmol) in MeOH (1 mL) was added a solution of 1 N HCI (1 mL) and the reaction mixture was heated at 60 °C for 2 h. All the volatiles were evaporated to give (±)-ery7zro-2-amino-3hydroxy-3-(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)butan-1 -one dihydrochloride Compound 239 (54 mg, 87 % yield) as a white solid.

Nh₂

2.HCI (±)

Compound 239

MW: 322.23; Yield: 87 %; White Solid; Mp (°C): 140.9

Ry: 0.1 (EtOAc:MeOH = 80:20, free base).

^-NMR (CDC1₃, δ): 1.85-2.05 (m, 7H, CH₃ + 2*CH₂), 3.35-3.65 (m, 4H, 2*NCH₂), 4.61 (s, 1H, O-CH), 8.23 (d, 2H, J= 4.5 Hz, ArH), 8.89 (d, 2H, J= 4.3 Hz, ArH).

Preparation of (5)-2-amino-3-(pyridin-4-yP-l-(pyrrolidin-l-vPpropan-l-one dihydrochloride Compound 240.

fe/Y-Butyl (5)- l-oxo-3-(pyridin-4-yP-1 -(pyrrolidin-1 -yl)propan-2-ylcarbamate EBE 06190.

To a solution of A-Boc-(25)-3-(pyridin-4-yl)alanine (500 mg, 1.9 mmol) in THF (12 mL) was added A-methylmorpholine (200 pL, 1.9 mmol) and the

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184 solution was stirred for 5 min, cooled at - 15 °C and treated dropwise with isobutyl chloroformate (249 pL, 1.9 mmol). The mixture was stirred for 10 min and pyrrolidine (1.08 g, 15.2 mmol) was added and allowed to warm to 25 °C with stirring for 3 h. The solvent were removed under reduced pressure and the residue was partitioned between EtOAc and NaH₂PO4 pH = 7.2. The aqueous layer was discarded and the organic layer was washed with aqueous saturated NaHCO3, dried over Na₂SO4, filtered and evaporated. The resulting solid was purified by column chromatography (SiO₂) with a gradient of 0 % to 10 % [v/v] MeOH in EtOAc to give tert-butyl (S)- l-oxo-3-(pyridin-4-yl)-1 -(pyrrolidin-1yl)propan-2-ylcarbamate EBE 06190 (167 mg, 28 % yield) as a white solid.

EBE 06190

MW: 319.4; Yield: 28 %; White Solid; Mp (°C): 130.0

R_f: 0.30 (EtOAciMeOH = 90:10).

^-NMR (CDC1₃, δ): 1.40 (s, 9H, C(CH₃)₃, 1.65-1.90 (m, 4H, CH₂), 2.80-2.95 (m, 1H, CH), 2.95-3.05 (m, 2H, CH₂), 3.30-3.45 (m, 1H, NCH), 3.45-3.55 (m, 2H, N-CH₂), 4.60-4.75 (m, 1H, N-CH), 5.42 (d, 1H, J= 8.8 Hz, NH), 7.16 (dd, 2H, .7= 4.5 Hz, 7= 1.5 Hz, ArH), 8.51 (dd, 2H,J= 1.5 Hz, J = 4.5 Hz, ArH). ¹³C-NMR (CDC1₃, δ): 24.1, 25.8,28.3 (3xC), 39.1,45.8,46.5, 52.6, 79.9, 124.8 (2xC), 145.7,149.8 (2xC), 155.0,169.2.

GS)-2-Amino-3-(pyridin-4-yl)-1-(pyrrolidin- l-yl)propan-1 -one dihydrochloride Compound 240.

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To a solution of ierLbutyl (5)-l-oxo-3-(pyridin-4-yl)-l-(pyrrolidin-lyl)propan-2-ylcarbamate EBE 06190 (100 mg, 0.313 mmol) in CH₂C1₂ (3 mL) at 4 °C was added TFA (479 pL, 6.26 mmol) and MeOH (0.3 mL) and the reaction was stirred for 2 h. All the volatiles were evaporated to give a product that was treated with a suspension of amberlite-400 (OH‘ form, 5 g) in MeOH. The suspension was filtered and washed with MeOH (5x5 mL). The combined methanol fractions were evaporated under reduced pressure and the desired product was isolated by column chromatography (SiO₂) with a gradient of 0 % to 30 % [v/v] MeOH in EtOAc to give (5)-2-amino-3-(pyridin-4-yl)-l(pyrrolidin-l-yl)propan-l-one. The product was dissolved in MeOH, cooled at 5 °C and a solution of HCI (0.1 N) (9 mL) was added dropwise. All the volatiles were evaporated to give (₍S)-2-amino-3-(pyridin-4-yl)-l-(pyrrolidin-l-yl)propan1-one dihydrochloride Compound 240 (91 mg, 99 % yield) as a white solid.

Compound 240

MW: 292.21; Yield: 99 %; White Solid; Mp (°C): 195.9

Ry: 0.10 (EtOAc:MeOH = 90: 10, free base).

Ή-NMR (CD₃OD, δ): 1.80-2.05 (m, 4H, CH₂), 3.30-3.40 (m, 5H, N-CH₂ + NCH), 3.60-3.75 (m, 1H, CH), 4.72 (t, 1H, J= Ί3 Hz, CH), 8.08 (d, 2H, J= 5.4 Hz, ArH), 8.87 (d, 2H, J= 5.3 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 25.0, 26.9, 37.2, 47.6, 48.0, 57.2, 130.0 (2xC), 143.1 (2xC), 157.3, 166.6.

MS-ESI m/z (% rel. Int.): 220.1 ([MH]⁺, 10), 203.1 (50).

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Preparation of(T)-//2reo-2-amino-3-(6-(trifluoromethvl)pyridin-3-vl)-3hydroxy-1-(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 241.

(±)-£rans-(5-(6-(Trifhioromethyl)pyridin-3-yl)-4,5-dihydrooxazol-4yl)(pyrrolidin-l-yl)methanone EBE 06196.

To a solution of KOH (184 mg, 3.28 mmol) in MeOH (10 mL) at 4 °C was added 6-(trifhioromethyl)pyridme-3-carbaldehyde (575 mg, 3.28 mmol) and and 2-isocyano-l-(pyrrolidin-l-yl)ethanone BLE 04098 (500 mg, 3.28 mmol). The mixture was stirred for 2 h at 4 °C and allowed to warm to 25 °C. All the volatiles were evaporated and the resulting product was partitioned between brine and EtOAc. The organic layer was separated, dried over Na₂SO₄, filtered to give after evaporation of the solvent (±)-/ra7t₁s^,-(5-(6(trifluoromethyl)pyridin-3-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-lyl)methanone EBE 06196 (801 mg, 78 % yield) as a pale brown solid.

(±)-trans

EBE 06196

MW: 310.13; Yield: 78 %; Pale brown solid.

Ή-NMR (CDC1₃, δ): 1.75-2.10 (m, 4H, 2xCH₂), 3.40-3.60 (m, 3H, 2*N-CH₂),

3.90-4.00 (m, 1H, N-CH), 4.57 (dd, 1H, J= 8.0 Hz, J = 2.2 Hz, N-CH), 6.31 (d, 1H, J= 8.0 Hz, O-CH), 7.06 (d, 1H, J = 2.2 Hz, N-CH), 7.71 (d, 1H, J= 8.1 Hz, ArH), 7.84 (dd, 1H, J= 1.8 Hz, J= 8.1 Hz, ArH), 8.70 (d, 1H, J- 1.5 Hz, ArH).

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187 (±)-/7zreo-2-Amino-3-(6-(trifluoromethyl)pvridin-3-yl)-3-hydroxy-l-(pyrrolidinl-yllpropan-l-one dihydrochloride Compound 241.

To a solution of (±)-/razz_>s'-(5-(6-(trifluoromethyl)pyridin-3-yl)-4₃5dihydrooxazol-4-yl)(pyrrolidin-l-yl)methanone EBE 06196 (400 mg, 1.28 mmol) in MeOH (2 mL) was added HCI (37 %) (10 mL) and the mixture was heated at 60 °C for 2 h with continuous stirring. After evaporation the resulting white solid was treated with a suspension of amberlite-400 (OH form) in MeOH. The suspension was filtered and washed with MeOH (5x5 mL). The combined methanol fraction were evaporated under reduced pressure and the desired product was isolated by column chromatography (SiO₂) with a gradient of 0 % to 8 % [v/v] MeOH in EtOAc to obtain (±)-/Zzreo-2-amino-3-(6(trifluoromethyl)pyridin-3-yl)-3-hydroxy-1 -(pyrrolidin-1 -yl)propan-1 -one (287 mg). To a solution (±)-//zreo-2-amino-3-(6-(trifluoromethyl)pyridin-3-yl)-3hydroxy-1-(pyrrolidin-l-yl)propan-l-one (209 mg, 0.69 mmol) in MeOH (2 mL) at 4 °C and treated HCI 37 % (10 mL). All the volatiles were evaporated to give (±)-/7zreo-2-amino-3-(6-(trifluoromethyl)pyridin-3-yl)-3-hydroxy-1 (pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 241 as a white solid (269 mg, 74 % yield).

OH 0 o

(±)

Compound 241

MW: 376.2; Yield: 74 %; White Solid; Mp (°C): 190.0.

Ry·: 0.3 (EtOAc:MeOH = 92:8, free base).

Ή-NMR (CD₃OD, δ): 1.40-1.70 (m, 2H, CH₂), 1.70-1.90 (m, 2H, CH₂), 2.352.50 (m, 1H, N-CH), 3.15-3.35 (m, 1H, N-CH₂), 3.35-3.45 (m, 2H, N-CH₂),

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4.32 (d, 1H, J= 8.3 Hz, N-CH), 5.11 (d, 1H, J= 8.2 Hz, O-CH), 7.88 (d, 1H, J = 8.3 Hz, ArH), 8.16 (d, 1H, J= 8.0 Hz, ArH), 8.73 (s, 1H, ArH).

¹³C-NMR (CD₃OD, δ): 24.8, 26.6, 47.4,47.9, 58.7, 71.3, 121.7, 122.9 (d, 1C, J = 273.2 Hz, CF₃), 137.9, 140.3, 149.0,149.7,165.8.

Preparation of (±)-Z7zreo-2-amino-3-hvdroxv-1 -(thiazolidin-3-yl)-3-(pyridin-4yl)propan-l-one dihydrochloride Compound 242.

(±)-//zreo-2-Amino-3-hydroxy-3-(pyridin-4-yl)propanoic acid dihydrochloride EBE 10038B.

To a stirred solution of KOH (2.57 g, 35.4 mmol) in MeOH (35 mL) at 0 °C was added 4-pyridinecarboxaldehyde (3.80 g, 35.4 mmol) and tertbutylisocyano acetate (5 g, 35.4 mmol). The solution was stirred for 3 h at 0 °C and concentrated to obtain intermediate iran5-4,5-dihydro-5-(pyridin-4yl)oxazole-4-carboxylate as a pale yellow solid EBE 10038A.

O^N

EBE 10038A

Ή NMR (CD₃OD, δ): 4.29 (dd, 1H, J= 1.7 Hz, J = 7.6 Hz, N-CH), 5.67 (d, 1H, J= 7.6 Hz, O-CH), 7.29 (d, 1H, J= 1.9 Hz, N=CH), 7.46 (d, 2H, J= 4.7 Hz, ArH), 8.55 (dd, 2H, J= 1.7 Hz, J= 4.4 Hz, ArH).

The solid EBE 1003 8A was dissolved in MeOH (100 mL), stirred 5 min at 0 °C and treated with HCI (12 N) (10.5 mL). The reaction was heated at 60 °C for 2 h, cooled down to 4 °C to form a precipitate that was filtered. The filtrate was evaporated and dried to obtain (±)-i/zreo-2-amino-3-hydroxy-3(pyridin-4-yl)propanoic acid dihydrochloride EBE 10038B with 20 % of (±)erythro isomer (9 g, 99 % crude yield) as a pale beige solid.

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EBE 10038B

MW: 255.10; Yield: 99 %; Pale Beige Solid.

^!H-NMR (CD₃OD, δ): 3.94 (d, 1H, 3.9 Hz, N-CH), 4.61 (t, 1H, J= 3.0 Hz,

O-CH), 8.31 (d, 2H, ArH), 8.95 (m, 2H, ArH). Only the formula and the ’HNMR description of major threo isomer is shown.

MS-ESI m/z (% rel. Int.): 183.1 ([MH]⁺, 5).

(±)-/Arg<?-W-Boc-2-amino-3-(pyridin-4-yl)-3-hydroxy-propionic acid EBE 10040.

A solution of di-terAbutyldicarbonate (9.28 g, 42.5 mmol) in dioxane (40 mL) was added to a pre-mixed ice cold solution of (±)-ZAreo-2-amino-3hydroxy-3-(pyridin-4-yl)propanoic acid dihydrochloride EBE 1OO38B (9.01 g,

35.4 mmol) in a solution of 1 N NaOH (145 mL). The biphasic mixture was stirred at 5 °C for 30 min and allowed to warm to room temperature for 3.5 h, concentrated, cooled in an ice bath, acidified to pH 4-5 and extracted with nbutanol. The combined extracts were dried over Na₂SO4 and filtered to give (±)i/zreo-WBoc-2-amino-3-(pyridin-4-yl)-3-hydroxy-propionic acid EBE 10040 (3.34g, 33 % yield) as a pale yellow solid.

OH 0

EBE 10040

MW: 282.2; Yield: 33 %; Pale Yellow Solid.

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190 ^-NMR (DMSO-d6, δ): 1.24 (s, 9H, C(CH₃)₃), 4.32 (d, 1H, J= 9.4 Hz, N-CH),

5.13 (s, 1H, O-CH), 6.35 (d, 1H, J= 9.4 Hz, NH), 7.37 (d, 2H, J= 3.8 Hz, ArH), 8.48 (d, 2H, J= 3.6 Hz), 2 OH not seen.

¹³C-NMR (DMSO-d6, δ): 27.9 (3xC), 59.0, 71.3, 78.2, 121.4, 122.8, 148.9,

150.3, 151.0, 155.2, 171.6.

MS-ESI m/z (% rel. Int.): 283.2 ([MH]⁺, 10).

HPLC: Method A, detection at 254 nm, EBE 10040 RT = 3.17 min, peak area

95.9 %.

tert-butyl (X)-threo-1 -hydroxy-3 -oxo-1 -(pyridin-4-yl)-3-(thiazolidin-3 yl)propan-2-ylcarbamate EBE 10042.

To a solution of (±)-iZzreo-7V-Boc-2-amino-3-(pyridin-4-yl)-3-hydroxypropionic acid EBE 10040 (500 mg, 1.77 mmol) in CH₂C1₂ (10 mL) at 0 °C was added triethylamine (253 pL, 3.54 mmol), hydroxybenzotriazole (239 mg, 1.77 mmol), l-(3-dimethylaminopropyl)-3-ethylcarbo-diimide hydrochloride (EDCI) (340 mg, 1.77 mmol) and thiazolidine (140 pL, 1.77 mmol). The reaction mixture was stirred for 2 h at 0 °C, allowed to warm to room temperature, stirred for 16 h and diluted in CH₂C1₂ (90 mL). The mixture was washed with brine (3x10 mL), 1 N NaOH (3x10 mL), dried over Na₂SO4, filtered to give a crude oil that was purified using column chromatography (SiO₂) with a gradient of 3 % to 4 % MeOH in CH₂C1₂. After evaporation tert-butyl (±)rt7zreo-lhydroxy-3-oxo-l-(pyridin-4-yl)-3-(thiazolidin-3-yl)propan-2-ylcarbamate EBE 10042 (200 mg, 32 % yield) was obtained as a yellow oil.

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OH O

N. / H(\l '-V

EBE 10042

MW: 353.4; Yield: 32 %; Yellow Oil.

R_z: 0.2 (CH₂Cl₂:MeOH = 97:3).

Ή-NMR (CDC1₃, δ): 1.29 (s, 9H, (CH₃)₃), 2.90-3.05 (m, 2H, S-CH₂), 3.60-3.95 (m, 2H, N-CH₂), 4.40-4.72 (m, 3H, N-CH + S-CH₂-N), 5.10-5.15 (m, 1H, OCH), 5.68 (m, 1H, NH), 7.34 (d, 2H, J= 6.0 Hz, ArH), 8.53 (d, 2H, J= 6.0 Hz, ArH), OH not seen.

¹³C-NMR (CDC1₃, δ), minor rotamer in parenthesis: 28.1 (29.2) [3xC], 31.1, (48.5) 48.7, 49.0 (49.1), (56.4) 56.7, 72.3, 80.5, 121.4 [2xC], 148.7, 149.4 [2xC], 155.5, (169.0) 169.2.

MS-ESI m/z (% rel. Int.): 354.2 ([MH]⁺, 20).

HPLC: Method A, detection at 254 nm, EBE 10042 RT = 3.87 min, peak area

98.1 %.

(±)-/Areo-2-Amino-3-hydroxv-l-(thiazolidin-3-yl)-3-(pyridin-4-yl)propan-l-one dihydrochloride Compound 242.

To a solution of tert-butyl (±)-/Are<2-l-hydroxy-3-oxo-l-(pyridin-4-yl)-3(thiazolidin-3-yl)propan-2-ylcarbamate EBE 10042 (150 mg, 424 mg) in MeOH (10 mL) at 4 °C was added a solution of 1 N HCI in MeOH (12 mL). The reaction mixture was allowed to warm at room temperature and stirred for 1 h. All the volatiles were evaporated to give an oily residue that was dissolved in MeOH and treated with EtOAc to form a precipitate. The volatiles were evaporated to give (±)-/Zzreo-2-amino-3-hydroxy-l-(thiazolidin-3-yl)-3-(pyridinWO 2006/081273

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4-yl)propan- 1-one dihydrochloride Compound 242 (135 mg, 98 % yield) as a pale yellow solid.

OH 0

V nh₂

2.HCI (±)

Compound 242

MW: 326.24; Yield: 98 %; Pale Yellow solid; Mp (°C): 210.6 Ή-NMR (CDC1₃, δ): 2.90-3.15 (m, 2H, S-CH₂), 3.55-3.90 (m, 1H, N-CH), 4.00-4.15 (m, 1H, N-CH), 4.18-4.53 (m, 1H, N-CH), 4.62-4.78 (m, 2H, N-CH₂S), 5.38-5.49 (m, 1H, O-CH), 8.25 (d, 2H, J= 5.7 Hz, 2H, ArH), 8.94 (d, 2H, J = 5.7 Hz, ArH).

¹³C-NMR (CDC1₃, δ), minor rotamer in parenthesis: (29.9) 31.8, 34.8, (51.1) 52.1, (57.8) 58.2, 71.0 (71.3), 126.8 [2xC], 143.2 [2xC], 161.4, (164.8) 165.2. MS-ESI m/z (% rel. Int.): 254.2 ([MH]⁺, 15).

Preparation of (±)-/7zreo-2-amino-3-hydroxy-1 -(indolin-l-yl)-3-(pvridin-4yl)propan-l-one dihydrochloride Compound 243.

To a solution of potassium trans-A,5-dihydro-5-(pyridin-4-yl)oxazole-4carboxylate EBE 10038A (500 mg, 2.60 mmol) in CH₂C1₂ (13 mL) were added HOBT (352 mg, 2.60 mmol), EDCI (499 mg, 2.60 mmol) and indoline (292 mL, 2.60 mmol). The reaction mixture was stirred 2 h at 0 °C, allowed to warm to RT and stirred for 16 h. The reaction mixture was diluted in CH₂C1₂ (100 mL), wash with brine (3x25 mL), NaOH IN (3x25 mL), dried over MgSO₄, filtered to give after evaporation ira/70^,-(4,5-dihydro-5-(pyridin-4-yl)oxazol-4yl)(17/-indol-l-yl)methanone SLA 09182 (533 mg, 70 % yield) as a pale brown oil. To a solution of SLA 09182 (533 mg, 2.6 mmol) in MeOH (12 mL) was

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193 added a solution of HCI 37 % (880 pL). The reaction was stirred for 3 h at 50 °C and concentrated under reduced pressure. The resulting product was dissolved in MeOH and treated with amberlite (OH’ form), filtered to give after evaporation a residue that was purified using silica gel chromatography with a gradient of MeOH 0 % -10 % in CH2CI2 to yield (±)-/Areo-2-amino-3-hydroxyl-(indolin-l-yl)-3-(pyridin-4-yl)propan-l-one. The hydrochloride salt was formed by treatment with a solution of IM HCI in MeOH (3.2 mL) to give after evaporation (±)-//ww-2-amino-3 -hydroxy-1 -(indolin-1 -yl)-3-(pyridin-4yl)propan-l-one dihydrochloride Compound 243 (149 mg, 16 % yield) as a white solid.

OH 0

HCI (±)

Compound 243

MW: 356.25; Yield: 16 %; White Solid; Mp (°C): 202.5

R_z: 0.30 (CH₂C1₂: MeOH = 90:10, free base)

Ή-NMR (CD3OD, δ): 3.65-3.80 (m, 1H, N-CH₂), 4.15-4.28 (m, 1H, CH₂), 3.65-3.80 (m, 1H, CH₂), 4.15-4.30 (m, 1H, N-CH₂), 4.64 (d, 1H, J= 5.4 Hz, NCH), 5.45 (d, 1H, J= 5.3 Hz, O-CH), 6.70 (t, 1H, J= 7.4 Hz, ArH), 7.12 (dd, 2H, J= Ί.Ί Hz, ArH), 8.07 (d, 1H, J= 7.9 Hz, ArH), 8.15 (d, 2H, J= 6.2 Hz, ArH), 8.79 (d, 2H, J= 6.2 Hz, ArH).

^,3C-NMR (CD3OD, δ): 28.9, 30.8, 58.4, 71.1, 118.5, 126.1, 126.4, 126.5 (2xC),

128.4,133.6, 142.9, 143.6 (2xC), 161.0, 164.7.

MS-ESI m/z (% rel. Int.): 284.2 ([MH]⁺, 10).

Preparation of (±)-/Areo-2-amino-3-(3,5-dichloropyridin-4-yl)-3-hydroxy-l(2#-pyrrol-l(577)-yl)propan-l-one dihydrochloride Compound 245.

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194 /F<7»5-(5-(3,5-Dichloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(277-pyrrol-l(5iZ)vDmethanone SLA 09022.

To a stirred and cooled (0 °C) solution of KOH (0.315 g, 5.62 mmol) in methanol (70 mL) was added a mixture of 3,5-dichloropyridine-4-carbaldehyde (0.989 mg, 5.62 mmol) and 2-isocyano-l-(2Z/'-pyrrol-l(5Z/)-yl)ethanone SLA 07178 (0.696 g, 5.11 mmol). The solution was stirred 2 h with continued cooling and then concentrated. The residue was partitioned between EtOAc (50 mL) and water (50 mL). The organic layer was combined with additional EtOAc extracts (3x50 mL), washed with brine (70 mL) and dried with MgSO4, filtered and evaporated to obtain a crude product which was purified by column chromatography (florisil, EtOAc:cyclohexane = 80:20) to obtain after evaporation iraz/s-(5-(3,5-dichloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(2Z/pyrrol-l(5//)-yl)methanone SLA 09022 (1.267 g, 80 % yield) as a pale yellow solid.

(+)-trans

SLA 09022

MW: 312.15; Yield: 80 %; Pale Yellow Solid.

Ry: 0.15 (EtOAc: cyclohexane = 80:20).

Ή-NMR (CDC1₃, δ): 4.30-4.32 (m, 3H, 1.5xCH₂), 4.77-4.83 (m, 1H, 0.5xCH₂), 4.86 (dd, 1H, J= 2.2 Hz, J= 8.8 Hz, CH-N), 5.84-5.88 (m, 2H, CH=CH), 6.86 (d, 1H, J= 8.7 Hz, CH-O), 6.96 (d, 1H, J= 2.2 Hz, O-CH=N), 8.52 (s, 2H, ArH).

¹³C-NMR (CDC1₃, δ): 52.1, 52.6, 59.2, 75.1, 123.9, 124.4, 131.5, 139.5, 147.6, 148.3,151.0, 153.9, 164.3.

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195 (±)-fAreo-2-Amino-3-(3,5-dichloropyridin-4-yl)-3-hydroxy- l-(27/-pyrrol-1 (577)vl)propan-l-one dihydrochloride Compound 245.

To a stirred solution of Zran5-(5-(3,5-dichloropyridin-4-yl)-4,5dihydrooxazol-4-yl)(277-pyrrol-l(577)-yl)methanone SLA 09022 (1.26 g, 4.04 mmol) in methanol (20 mL) was added HCI 37 % (1.5 mL). The reaction mixture was stirred for 3 h at RT, concentrated and the resulting yellow oil was co evaporated twice with EtOAc and triturated with EtOAc to obtain after filtration and drying under vacuum (±)-/Areo-2-amino-3-(3,5-dichloropyridin-4yl)-3-hydroxy-l-(277-pyrrol-l(577)-yl)propan-l-one dihydrochloride (1.13 g, 75 % yield) as a pale yellow solid.

oh o

Cl

2HCI (±)

Compound 245

MW: 375.16; Yield: 75 %; Pale yellow solid; Mp (°C): 176.5

Ry. 0.15 (CH₂C1₂: MeOH = 90:10, free base).

Ή-NMR (CD₃OD, δ): 3.30-3.32 (m, 1H, 0.5xCH₂), 3.90-3.97 (m, 1H, 0.5xCH₂), 4.25-4.37 (m, 2H, CH₂), 4.76 (d, 1H, J= 10.3 Hz, N-CH), 5.60-5.65 (m, 1H, =CH), 5.70 (d, 1H, J= 10.3 Hz, O-CH) 5.78-5.82 (m, 1H, =CH), 8.57 (s, 2H, ArH).

¹³C-NMR (CD₃OD, δ): 56.6, 56.7, 57.3, 72.4, 128.0, 128.9, 136.3(2C), 145.8, 152.5(2C), 168.1.

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Preparation of N-( 1 -oxo-3-(pyridin-4-yl)-1 -(pyrrolidin-1 -vl)prop-2-en-2yl)formamide hydrochloride (cis:trans isomers mixture) Compound 246.

To a stirred solution of triphenylphosphine (200 mg, 0.76 mmol) in 5 mL of CH₃CN were added at 20 °C diethylazodicarbonate (120 pl, 0.76 mmol), Et₃N (55 pl, 0.38 mmol) and7V'-((±)-iAreo-l-hydroxy-3-oxo-l-(pyridin-4-yl)-3(pyrrolidin-l-yl)propan-2-yl)formamide hydrochloride Compound 216 (115 mg, 0.38 mmol). The mixture was stirred 2 h at 70 °C then solvent was evaporated. The obtained residue was purified by column chromatography (SiO₂, EtOAc:MeOH = 9:1) to give 7V-(l-oxo-3-(pyridin-4-yl)-l-(pyrrolidin-l-yl)prop-

2-en-2-yl)formamide TTA 08074A (80 mg, 43 % yield). HCI treatment in EtOAc with HCI 0.4 N in diethyl ether (1 mL, 0.4 mmol) gave after evaporation and drying N-( 1 -oxo-3 -(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)prop-2-en-2yl)formamide hydrochloride (trans'.cis isomers mixture) Compound 246 (60 mg, 28 % yield) as a pale yellow pasty product.

HCI cis:trans mixture

Compound 246

MW: 281.74; Yield: 28 %; Pale Yellow Pasty Product.

R_f: 0.24 (EtOAc:MeOH = 9:1).

Ή-NMR (CD₃OD, δ): 1.80-2.05 (bs, 4H, 2xCH₂), 3.25-3.45 (bs, 2H, CH₂-N),

3.55-3.70 (bs, 2H, CH₂-N), 6.87 (s, 1H, CH), 7.82 (d, 2H, J= 5.0 Hz, ArH),

8.31 (s, 1H, HC=O), 8.65 (d, 2H, 7= 5.0 Hz, ArH).

¹³C-NMR(CD₃OD, δ): 25.1,26.5, 47.1,48.9,109.0,125.6 (2xC), 127.7, 142.2 (2xC), 142.6, 154.9, 161.6.

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MS-ESI m/z (% rel. Int.): 246.1 ([MH]⁺, 5), 175.1 (100).

HPLC: Method A, detection UV 254 nm, Compound 246 RT = 1.90 min, peak area 95.0 %.

Preparation of (2S,3R)~ & (2R,35)-2-amino-l-i(R)-3-hydroxypyrrolidin-l-yl)-3hydroxy-3-(pyridin-4-yl)propan-l-one dihydrochlorides Compound 247.

l-((R)-3-hydroxypyrrolidin-l-yl)-2-isocyanoethanone VIB 01172.

To stirred and cooled (0 °C) methyl isocyanoacetate (96 % technical grade, 1.7 g, 17.21 mmol) was slowly added (7?)-(+)-3-pyrrolidinol (1.5 g, 17.21 mmol) and MeOH (5 mL). The mixture was stirred for 3 h at RT and concentrated. Brine was added (30 mL) and the mixture was extracted with EtOAc (3x50 mL), dried over MgSO₄, filtered and evaporated to obtained crude l-((R)-3-hydroxypyrrolidin-l-yl)-2-isocyanoethanone VIB 01172 (1.3 g, 49 % yield) as a yellow solid.

VIB 01172

MW: 154.17; Yield: 49 %; Yellow Solid; Mp (°C) = 55.9 ^!H-NMR (CDC1₃, δ): 1.95-2.20 (m, 2H, CH₂), 2.60-2.82 (m, 1H, OH), 3.30-

3.68 (m, 4H, 2xCH₂), 4.20-4.36 (m, 2H, CH₂), 4.49-4.65 (m, 1H, CH-O). MS-ESI m/z (% rel. Int.): 155.1 ([MH]⁺, 90).

/r6OTs-((R)-3-hydroxypyrrolidin-l-ylX(45,5R)- & (4R,55)-4,5-dihydro-5(pyridin-4-yl)oxazol-4-yl)methanones VIB 01174.

To a stirred and cooled (0 °C) solution of KOH (0.40 g, 7.13 mmol) in

MeOH (8 mL) were added successively pyridine-4-carbaldehyde (0.84 g, 7.84

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198 mmol) and l-((R)-3-hydroxypyrrolidin-l-yl)-2-isocyanoethanone VIB 01172 (1.10 g, 7.13 mmol). The mixture was stirred at 0 °C to RT for 24 h. After evaporation of MeOH, the mixture was partitioned between EtOAc (50 mL) and H₂O (10 mL). The aqueous layer was further extracted with EtOAc (2x50 mL).

The EtOAc fractions were combined, washed twice with brine (2x10 mL), dried over MgSO4, filtered and evaporated. After evaporation and drying iran.s-((R)-

3-hydroxypyrrolidin-l-yl)((4₍S',5R)- & (4R,50)-4,5-dihydro-5-(pyridin-4yl)oxazol-4-yl)methanones VIB 01174 (490 mg, diastereoisomeric mixture in ratio 1:1, 26 % yield) were obtained as a crude pale yellow solid.

VIB 01174

MW: 261.28; Yield: 26 %; Pale Yellow Solid.

Ή NMR (CDC1₃, δ): 1.88-2.22 (m, 2H, CH₂), 3.50-3.80 (m, 3H, 1.5xCH₂),

3.95-4.20 (m, 1H, 0.5xCH₂), 4.40-4.65 (m, 2H, CH-N & CH-O), 4.74 (s, 1H,

OH), 6.18-6.22 (m, 1H, CH-O), 7.00-7.12 (m, 1H, HC=N), 7.20-7.30 (m, 2H,

ArH), 8.52-8.68 (m, 2H, ArH).

MS-ESI m/z (% rel. Int.): 262.2 ([MH]⁺, 45), 235.2 (75), 148 (100).

(2S,3R)~ & (2R,35)-2-Amino-l-((R)-3-hvdroxypyrrolidin-l-yl)-3-hydroxy-3(pyridin-4-yl)propan-l-one dihydrochlorides Compound 247.

To a solution of iraw5-((R)-3-hydroxypyrrolidin-l-yl)((4₁S',5R)- & (4R,5S)~

4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)methanones VIB 01174 (0.49 g, 1.87 mmol) in methanol (6.5 mL) was added hydrochloric acid 37 % (575 pL). After heating (50 °C) the mixture for 2 h the reaction mixture was concentrated and

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199 the crude product was coevaporated twice with EtOAc. After trituration with

EtOAc, filtration and drying (2S,3R)- & (2R,36)-2-amino-l-((7?)-3hydroxypyrrolidin-1 -yl)-3-hydroxy-3-(pyridin-4-yl)propan-1 -one dihydrochlorides (420 mg, diastereoisomeric mixture in ratio 1:1, 69 % yield) were obtained as a pale pink solid.

Compound 247

MW: 324.2; Yield: 69 %; Pale Pink Solid; Mp (°C): 177.0

Ή-NMR (CD3OD, δ): 1.88-2.22 (m, 2H, CH₂), 2.70-3.80 (m, 4H, 2xCH₂),

4.20-4.65 (m, 2H, CH-O & CH-N), 5.20-5.45 (m, 1H, CH-O), 8.10-8.25 (m, 2H, ArH), 8.80-9.00 (m, 2H, ArH), 2xOH & NH₂ not seen.

MS-ESI m/z (% rel. Int.): 252.2 ([MH]⁺, 37), 235.1 (63), 148.0 (100).

Preparation of i-)-//zreo-2-amino-1 -((6)-3 -fluoropyrrolidin-1 -yl)-3 -hydroxy-3 (pyridin-4-yDpropan-l-one dihydrochloride Compound 248 and (+)-threo-2amino-1 -((6)-3-fluoropyrrolidin-1 -yl)-3-hydroxy-3-(pyridin-4-yl)propan-1 -one dihydrochloride Compound 249.

Extraction of the free base:

(2S,3R)- & (2R,36)-2-amino-l-((6)-3-fluoropyrrolidin-l-yl)-3-hydroxy-3(pyridin-4-yl)propan-l-one dihydrochlorides Compound 214 (300 mg, 0.92 mmol) were dissolved in 10 mL of a Na₂CO3 (10 %) solution and the aqueous mixture was then saturated with NaCl. The aqueous phase was extracted by 5 x

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200 mL of a mixture CH₂Cl₂:2-PrOH (9:1). The organic phase was dried over MgSO₄ and evaporated to afford 163 mg (70 %) of the corresponding free base of Compound 214.

Analytical chiral separation:

pL of a 1 mg/mL solution of Compound 214 were injected on Chiralpak AD: flow-rate = 1 mL/min, temperature = 25 °C, mobile phase hexane:ethanol = 7:3, detection on UV 220 nm and on polarimeter, first eluted diastereoisomer Compound 248 Rtl(-) = 20.94 min, second eluted diastereoisomer Rt2(+) = 24.77 min, kl(-) = 5.93, k2(+) = 7.20, a = 1.21 and resolution Rs = 1.21. The integration of the UV signal gives 42 % for the first diastereoisomer compound 248 and 58 % for the second Compound 249 (the UV response is different for the two diastereoisomers).

Semi-preparative chiral separation:

170 mg of the free base of Compound 214 were dissolved in 6 mL of ethanol, and 30 pL of this solution were injected every 9 min on Chiralpak ADH, flow-rate = 2 mL/min, mobile phase hexane:ethanol = 7:3, detection on UV 220 nm. 195 successive injections were done. The two main fractions were identified on UV and collected in two different flasks. The solvent was removed in vacuo at 30 °C. The resulting solid was dissolved in 50 mL of CH₂C1₂ and then filtered on a 0.45 pm millipore membrane. After evaporation of CH₂C1₂, the solid was dissolved in 50 mL of methanol and then filtered. For the free base of the first diastereoisomer, a new series of injections was needed to remove two UV-visible impurities collected in the same flask: in the same chromatographic conditions, 30 injections of 100 pL of a 25 mg/mL solution were made every 20 min. The salts Compound 248 and Compound 249 were

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201 regenerated according to the same procedure reported for Compound 203 and Compound 204.

The regenerated salts Compound 248 and Compound 249 were injected in analytical conditions, the diastereoisomeric excesses for Compound 248 and Compound 249 were determined to be higher than 96 %.

(-)-/Areo-2-Amino-1 -((S)-3-fhioropyrrolidin-1 -yl)-3-hydroxy-3-(pyridin-4yPpropan-l-one dihydrochloride Compound 248.

OH 0

AZy ^NA nA ^nhz

2.HCI (-)-fhreo

Compound 248

MW: 326.19; 70 mg obtained; Pale Yellow Solid; Mp (°C): too hygroscopic. Diastereoisomeric excess > 96 % measured by HPLC at 220 nm (Chiralpak AD).

a²⁵ _D = - 2.0 (methanol, c = 1).

Ή-NMR (CD₃OD, δ): 1.85-2.38 (m, 2H, CH₂), 2.72-4.05 (m, 4H, 2xCH₂), 4.49-4.62 (m, 1H, CH'N), 5.10-5.48 (m, 2H, CH-O & CH-F), 8.11-8.25 (m, 2H, ArH), 8.82-8.98 (m, 2H, ArH).

(+)-//zreo-2-Amino-1 -((5)-3 -fluoropyrrolidin-1 -yp-3-hydroxy-3 -(pyridin-4yl)propan-l-one dihydrochloride Compound 249.

OH O

II Ί i 7 r nA ^nh2 —'^(S)

2.HCI (+)-threo

Compound 249

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202

MW: 326.19; 85 mg obtained; Pale Yellow Solid; Mp (°C): too hygroscopic. Diastereoisomeric excess > 96 % measured by HPLC at 220 nm (Chiralpak AD).

a²⁵ _D = + 31.7 (methanol, c = 1).

^-NMR (CD₃OD, δ): 1.82-2.38 (m, 2H, CH₂), 2.90-4.00 (m, 4H, 2xCH₂), 4.35-4.60 (m, 1H, CH'N), 5.00-5.48 (m, 2H, CH-0 & CH-F), 8.11-8.25 (m, 2H, ArH), 8.82-9.00 (m, 2H, ArH).

Preparation of (+)-//?reo-2-Amino-7V-ethyl-3-hydroxy-7V-methvl-3-pyridin-4-ylpropanamide dihydrochloride Compound 250.

fraws-WethyI-4,5-dihydro-Wmethyl-5-(pyridin-4-yr)oxazole-4-carboxamide SLA 09190.

To a solution of potassium iran5-4,5-dihydro-5-(pyridin-4-yl)oxazole-4carboxylate EBE 10038A (501 mg, 2.60 mmol) in CH₂C1₂ (12 mL) were added HOBT (352 mg, 2.60 mmol), EDCI (500 mg, 2.60 mmol) and Nmethylethanamine (223 mL, 2.60 mmol). The reaction mixture was stirred 2 h at 0 °C, allowed to warm to room temperature and stirred for 16 h. The reaction mixture was diluted in CH₂C1₂ (100 mL), washed with brine (2x25 mL), 1 N NaOH (2x25 mL), dried over MgSO₄, filtered to give after evaporation trans-Nethyl-4,5-dihydro-A^r-methyl-5-(pyridin-4-yl)oxazole-4-carboxamide SLA 09190 (144 mg, 24 % yield) as a pale brown oil.

(+)-trans

SLA 09190

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203

MW: 233.27; Yield: 24 %; Pale Brown Oil.

Ή NMR (CDC1₃, δ): 1.15-1.30 (m, 3H, CH₃), 3.22 (s, 3H, CH₃-N), 3.40-3.80 (m, 2H, N-CH₂), 4.59 (dd, 1H, J= 2.2 Hz, J= 7.8 Hz, N-CH), 6.24 (d, J= 7.7 Hz, OCH), 7.02 (d, 1H, J= 1.0 Hz, N=CH), 7.23 (d, 2H, J= 4.8 Hz, ArH), 8.61 (d, 2H, 4.6 Hz, ArH).

MS-ESI m/z (% rel. Int.): 234.2 ([MH]⁺, 30).

(±)-^reo-2-Amino-A^r-ethyl-3-hydroxy-7V-methyl-3-pyridin-4-yl-propanamide dihydrochloride Compound 250.

To a solution of 7V-ethyl-4,5-dihydro-/V-methyl-5-(pyridin-4-yl)oxazole-4carboxamide SLA 09190 (144 mg, 0.6 mmol) in MeOH (5 mL) was added a solution of HCI 37 % (240 pL). The reaction was stirred for 3 h at 50 °C and concentrated under reduced pressure. The resulting product was dissolved in MeOH and treated with amberlite (OH form), filtered to give after evaporation a residue that was purified by column chromatography (SiO₂, with a gradient of MeOH 10 % in CH₂C1₂) to yield to 2-amino-7V-ethyI-3-hydroxy-7V-methyl-3pyridin-4-yl-propionamide. The hydrochloride salt was formed by treatment of this free base with a solution of HCI 1 M in MeOH (1 mL) to give after evaporation 2-amino-7V-ethyl-3-hydroxy-7V-methyl-3-pyridin-4-yl-propanamide dihydrochloride Compound 250 as a pale yellow solid (80 mg, 44 % yield).

Compound 250

MW: 296.19; Yield: 44 %; Pale Yellow Solid; Mp (°C): 114.7 R_z: 0.30 (CH₂Cl₂:MeOH = 90:10, ffeebase).

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Ή-NMR (CD₃OD, δ): 0.96-1.10 (m, 3H, CH₃), 2.81 (s, 1.8H major rotamer, 0.6xCH₃), 2.88 (s, 1.2H minor rotamer, 0.4CH₃), 3.18-3.55 (m, 2H, CH₂), 4.66 (d, 0.4H minor rotamer, J= 6.6 Hz, 0.4xN-CH), 4.69 (d, 0.6H major rotamer, J = 6.3 Hz, 0.6xN-CH), 5.24 (d, 0.4H minor rotamer, J= 6.9 Hz, 0.4xO-CH), 5.27 (d, 0.6H major rotamer, J= 6.3 Hz, 0.6xO-CH), 8.08 (t, 2H, J= 6.5 Hz, ArH), 8.86 (d, 2H, J= 5.0 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 12.0, (13.6), (33.4), 35.3, 44.5, (45.6), 56.1, (56.2), 71.6, (72.1), 126.2 (2*C), 144.5, 144.6, 159.6, 166.5. () Minor rotamer in parenthesis.

Preparation of (2R,36>2-(3,4-dichlorobenzylamino)-3-hydroxy-3-(pyridin-4yl)-l-(pyrrolidin-l-vl)propan-l-one dihydrochloride Compound 251.

To a solution of (-)-(2R,3jS)-2-amino-3-hydroxy-3-(pyridin-4-yl)-l(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 203 (175 mg, 0.57 mmol) and Et₃N (175 pL, 1.25 mmol) in MeOH (5 mL) in a 50 mL round bottom flask equipped with a magnetic stirrer and under nitrogen atmosphere was added slowly at RT 3,4-dichlorobenzaldehyde (112 mg, 0.63 mmol). The reaction mixture was stirred at RT for 20 h. Then AcOH (65 pL, 1.15 mmol) and NaBH₃CN (50 mg, 0.74 mmol) were added. The reaction mixture was stirred at RT for another 15 h. MeOH was evaporated and EtOAc (100 mL) was added. The organic phase was washed with a mixture of saturated sodium carbonate (5 mL) and brine (20 mL), then with brine (10 mL) and dried over MgSO₄, filtered and evaporated. The crude product was purified by column chromatography (SiO₂, eluent EtOAc:MeOH = 95:5) to give an oil (-)-(2R,3»S)-

2-amino-3 -hydroxy-3 -(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)propan-1 -one (182 mg, 81 % yield). This free base (182 mg, 0.46 mmol) was dissolved in MeOH (2 mL) at 4 °C and a solution of HCI 0.1 N in isopropanol (10.2 mL, 1.01 mmol)

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205 was added. After evaporation at 30 °C, a mixture of EtOAc:MeOH = 95:5 was added to yield, after evaporation and drying, to (27?,35)-2-(3,4dichlorobenzylamino)-3 -hydroxy-3 -(pyridin-4-yl)-1 -(pyrrolidin-1 -yl)propan-1 one dihydrochloride Compound 251 as a white solid (208 mg, 78 % yield).

OH o

Cl

Compound 251

MW: 467.22; Yield: 78 %; White Solid; Mp (°C): 195.1

Ry: 0.22 (EtOAc:MeOH = 95:5, free base).

Ή-NMR (CD₃OD, δ): 1.55-1.77 (m, 4H, 2xCH₂), 2.46-2.53 (m, 1H, 0.5xNCH₂), 3.20-3.30 (m, 3H, 1.5xN-CH₂), 4.25 (d, 1H, 7= 13.3 Hz, 0.5xN-CH₂), 4.38 (d, 1H, 7= 13.3 Hz, 0.5xN-CH₂), 4.52 (d, 1H, 7= Ί.Ί Hz, N-CH), 5.33 (d, 1H, 7= 7.7 Hz, O-CH), 7.48 (dd, 1H, 7= 8.3 Hz, 7= 1.7 Hz, ArH), 7.62 (dd, 1H, 7= 8.3 Hz, 7= 1.2 Hz, ArH), 7.74 (s, 1H, ArH), 8.15 (d, 2H, 7= 5.6 Hz, ArH), 8.91 (d, 2H, 7= 5.6 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 24.7,26.6,47.6,48.2, 50.4, 64.0, 72.2, 126.8 (2xC),

131.7, 132.2, 132.3, 133.8, 133.9, 135.1, 143.5 (2xC), 160.6, 163.8.

MS-ESI m/z (% rel. Int.): 394.1/396.1/398.1 ([MH]⁺, 60/45/10), 219.2 (100). HPLC: Method A, detection UV 254 nm, Compound 251 RT = 3.83 min, peak area 99.5 %.

Preparation of (25^,,37?)-2-(3,4-dichlorobenzylamino)-3-hydroxy-3-(pyridin-4yl)-l-(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 252.

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206

Same experimental as forCompound 251 preparation starting from (+)(25,3R)-2-amino-3-hydroxy-3-(pyridin-4-yl)-l-(pyrrolidin-l-yl)propan-l-one dihydrochloride Compound 204 (175 mg, 0.57 mmol). After purification by column chromatography (25,3/?)-2-(3,4-dichlorobenzylamino)-3 -hydroxy-3 (pyridin-4-yl)-l-(pyrrolidin-l-yl)propan-l-one was obtained (187 mg, 83 % yield). This free base (187 mg, 0.47 mmol) was dissolved in MeOH (2 mL) at 4 °C and a solution of HCI 0.1 N in isopropanol (10.4 mL, 1.04 mmol) was added. After evaporation at 30 °C, a mixture of EtOAc:MeOH = 95:5 was added to yield, after evaporation and drying, to Compound 252 as a white solid (212 mg, % yield).

oh o ci

Compound 252

MW: 467.22; Yield: 80 %; White Solid; Mp (°C): 187.5

Ry: 0.22 (EtOAc:MeOH = 95:5, free base).

Ή-NMR (CDC1₃, δ): 1.55-1.77 (m, 4H, 2xCH₂), 2.46-2.53 (m, 1H, 0.5xN-CH₂),

3.20-3.30 (m, 3H, 1.5xN-CH₂), 4.25 (dd, 1H, J= 13.3 Hz, 0.5xN-CH₂), 4.40 (d, 1H, J= 13.3 Hz, 0.5xN-CH₂), 4.52 (d, 1H, J= 7.7 Hz, N-CH), 5.33 (d, 1H, J=

7.4 Hz, O-CH), 7.48 (d, 1H, J= 8.3 Hz, ArH), 7.62 (dd, 1H, J= 8.3 Hz, J= 0.8 Hz, ArH), 7.74 (s, 1H, ArH), 8.15 (d, 2H, J= 5.7 Hz, ArH) 8.91 (d, 2H, J= 5.7 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 24.7, 26.6, 47.6,48.2, 50.4, 64.0, 72.3, 126.8 (2xC),

131.7,132.2,132.3, 133.8, 133.9,135.1,143.5 (2xC), 160.6,163.8.

MS-ESI m/z (% rel. Int.): 394.1/396.1/398.1 ([MH]⁺, 60/45/10), 219.2 (100).

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HPLC: Method A, detection UV 254 nm, Compound 252 RT = 3.83 min, peak area 99.5 %.

Preparation of (£L3-(pyridin-4-yl)-l-(pyrrolidin-l-yl)prop-2-en-l-one hydrochloride Compound 253.

To a solution of 3-(4-pyridinyl)acrylic acid (1.01 g, 6.77 mmol) in CHCI3 (20 mL) in a 100 mL round bottom flask equipped with a magnetic stirrer and under nitrogen atmosphere was added 1-hydroxybenzotriazole (1.11 g, 8.21 mmol). The reaction mixture was stirred at RT for 10 min. Then 1-(3dimethylaminopropyl)-3-ethylcarbodiimide (1.56 g, 8.15 mmol) was added. The reaction mixture was stirred at 4 °C for 10 min. Then pyrrolidine (1.11 mL, 18.3 mmol) was added slowly and the reaction mixture was stirred for 15 h at +4 °C to RT. Dichloromethane (200 mL) was added and organic phase was washed with brine (100 mL), a solution of NaOH 0.5 N (100 mL) and brine (50 mL). The organic phase was dried over MgSO4, filtered, and evaporated to obtain (£)-3-(pyridin-4-yl)-l-(pyrrolidin-l-yl)prop-2-en-l-one (1.30 g, 94 % yield). This free base (1.3 g, 6.40 mmol) was dissolved in MeOH (10 mL) at 4 °C and a solution of HCI 0.1 N in isopropanol (79 mL, 7.9 mmol) was added. After evaporation at 30 °C and drying, (£)-3-(pyridin-4-yl)-l-(pyrrolidin- l-yl)prop-2en-l-one hydrochloride Compound 253 was obtained as a beige solid (1.40 g, 87 % yield).

Compound 253

MW: 238.71; Yield: 87 %; Beige Solid; Mp (°C): 229.4

Ry: 0.35 (EtOAc : MeOH = 95:5, free base).

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Ή-NMR (CD₃OD, δ): 1.92-2.10 (m, 4H, 2xCH₂), 3.56 (t, 2H, 6.7 Hz, NCH₂), 3.80 (t, 2H, 6.7 Hz, N-CH₂), 7.59 (d, 1H, J= 15.6 Hz, CH=C), 7.68 (d, 1H, J= 15.6 Hz, CH=C), 8.35 (d, 2H, J-5.7 Hz, ArH) 8.86 (d, 2H, J= 5.6 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 25.2, 27.0,47.6,48.2,126.7 (2xC), 131.9, 136.5, 143.1 (2xC), 154.5, 164.6.

MS-ESI m/z (% rel. Int.): 203.2 ([MH]⁺, 100).

HPLC: Method A, detection UV 254 nm, Compound 253 RT = 3.18 min, peak area 99.5 %.

Preparation of (±)-/Areo-2-amino-3 -hydroxy-3 -(1 -methyl- 177-imidazol-2-yP-1 (pyrrolidin-1-yppropan-1-one dihydrochloride Compound 254. teflns-(4,5-Dihydro-5-( 1 -methyl- 177-imidazol-2-vPoxazol-4-yP(pyrrolidin-1 yDmethanone LPQ 01190B.

To a stirred and cooled (0 °C) solution of potassium hydroxide (0.33 g, 5.0 mmol) in MeOH (6 mL) were added a mixture of l-methyl-2-imidazole carboxaldehyde (0.56 g, 5.0 mmol) and 2-isocyano-l-(pyrrolidin-l-yl)ethanone BLE 04098 (0.70 g, 5.0 mmol). The solution was stirred 3 h at 4 °C and then concentrated. The residue was partitioned between EtOAc (100 mL) and water (20 mL). The organic layer was washed with brine (10 mL) and dried over MgSO₄, filtered and evaporated. Concentration afforded a crude product which was purified by column chromatography (florisil, EtOAc:MeOH = 95:5 to 90:10) to yield, after evaporation and drying, to 7ran₎s'-(4,5-dihydro-5-(l-methyl17Z-imidazol-2-yl)oxazol-4-yl)(pyrrolidin-l-yl)methanone LPO 01190B (0.32 g, 25 % yield) as a brown oil.

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209 (+)-trans

LP0 01190B

MW: 248.28; Yield: 25 %; Brown Oil. Ry·: 0.30 (EtOAc:MeOH = 9:1, free base).

Ή-NMR (CDC1₃, δ): 1.94-2.12 (m, 4H, 2xCH₂), 3.50 (t, 2H, J= 6.5 Hz, NCH₂), 3.69-4.13 (m, 5H, N- CH₂ ,N-CH₃), 5.68 (dd, 1H, J= 7.7 Hz, J= 2.3 Hz, CH-N), 6.19 (d, 1H, J= 7.7 Hz, CH-O), 6.82 (d, 1H, J= 2.2 Hz, CH=N), 6.94 (d, 1H, J= 1.1 Hz, ArH), 7.00 (d, 1H, J= 1.1 Hz, ArH).

¹³C-NMR (CDC1₃, δ): 24.2,26.0, 32.9,46.4,46.7, 71.2, 72.7,123.0,127.9,

143.7,153.7, 166.6.

MS-ESI m/z (% rel. Int.): 267.3 ([MH+18]⁺, 10), 196.2 (100).

HPLC: Method A, detection UV 254 nm, LPO 01190B RT = 3.92 min, peak area 99.5 %.

(±)-Z/zz-go-2-Amino-3 -hydroxy-3-( 1 -methyl- l/Z-imidazol-2-yl)-1 -(pyrrolidin-1 yl)propan-l-one dihydrochloride Compound 254.

A solution of trans-(A,5-dihydro-5-(l-methyl-177^r-imidazol-2-yl)oxazol-4yl)(pyrrolidin-l-yl)methanone LPO 01190B (320 mg, 1.29 mmol) and HCI 37 % (0.4 mL, 13 mmol) in MeOH (6 mL) was stirred at 50 °C for 3 h in a 50 mL round bottom flask. The solvent was evaporated and the product was precipitated by a mixture of MeOH:EtOAc:Et₂O = 3:12:5 (20 mL). Solvents were evaporated at 30 °C to give, after evaporation and drying, (±)-i/zreo-2amino-3-hydroxy-3-( 1 -methyl- lH-imidazol-2-yl)-1 -(pyrrolidin-1 -yl)propan-1 WO 2006/081273

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210 one dihydrochloride Compound 254 as a pale yellow solid (305 mg, 76 % yield).

OH O

2.HCI (+)-threo

Compound 254

MW: 311.21; Yield: 76 %; Pale Yellow Solid; Mp (°C): 183.4

R_z: 0.30 (CH₂Cl₂:MeOH = 95:5, free base).

^!H-NMR (CD₃OD, δ): 1.75-1.99 (m, 4H, 2xCH₂), 2.80-2.88 (m, 1H, 0.5xNCH₂), 3.30-3.70 (m, 3H, 1.5xN-CH₂), 3.95 (s, 3H, N-CH₃), 4.62 (d, 1H, J= 8.3 Hz, N-CH), 5.51 (d, 1H, J= 8.3 Hz, O-CH), 7.66 (s, 2H, ArH).

¹³C-NMR (CD₃OD, δ): 24.9, 26.9, 36.0,47.9 (2xC), 56.2, 65.1, 121.1, 126.5, 145.1, 164.4.

MS-ESI m/z (% rel. Int.): 239.3 ([MH]⁺, 10), 134.1 (100).

HPLC: Method A, detection UV 254 nm, RT = 0.8 min, peak area 99.5 %.

Preparation of (2Y3R)- & (2R,36)-2-amino-l-((6>3-fluoropyrrolidin-l-yl)-3hydroxy-3-(pyridin-3-yl)propan-l-one dihydrochlorides Compound 255.

frans-((SY3-fhioropyrrolidin- l-yl)((4£.,5R)- & (4R,5S)-4,5-dihydro-5-(pyridin-

4-yl)oxazol-4-yl)methanones SLA 11014.

To a stirred and cooled 0 °C solution of KOH (0.216 mg, 4.22 mmol) in methanol (5 mL) was added l-((«S)-3-fluoropyrrolidin-l-yl)-2-isocyanoethanone VIB 01166 (0.600 g, 4.22 mmol) and pyridine-3-carbaldehyde (0.40 mL, 3.84 mmol). The solution was stirred for 20 h at 0 °C. After evaporation under reduced pressure, the residue obtained was partitioned between EtOAc and

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H₂O. The product was extracted with EtOAc (4x50 mL) and washed with brine (25 mL), dried over MgSO₄, filtered and evaporated to yield to a product that was purified using chromatography (florisil, EtOAc:MeOH = 95:5), ^/75-((5)-

3- fluoropyrrolidin- l-yl)((45',57?)- & (47?,55)-4,5-dihydro-5-(pyridin-4-yl)oxazol-

4- yl)methanones SLA 11014 were obtained as a yellow solid (0.464 g, diastereoisomeric mixture in ratio about 1:1,46 % yield).

trans trans

SLA 11014

MW: 263.27; Yield: 46 %; Yellow Solid; Mp (°C) = 171.7

R/ 0.25 (EtOAc:MeOH = 95:5).

Ή-NMR (CDC1₃, δ): 1.85-2.45 (m, 2H, CH₂), 3.50-4.10 (m, 3H, CH₂ & N-CH),

4.25-4.65 (m, 2H, N-CH₂), 5.15-5.25 (m, 0.5H, 0.5xCHF), 5.35-5.45 (m, 0.5H, 0.5xCHF), 6.21 (d, 1H, J= 7.6 Hz, O-CH), 7.04 (d, 1H, J= 2.1 Hz, N=CH),

7.30-7.38 (m, 1H, ArH), 7.60-7.68 (m, 1H, ArH), 8.55-8.65 (m, 2H, ArH). MS-ESI m/z (% rel. Int.): 264.1 ([MH]⁺, 18).

(25,37?)- & (27?,35)-2-Amino-1 -((5)-3-fluoropyrrolidin-1 -yl)-3-hydroxy-3(pyridin-3-yl)propan-l-one dihydrochlorides Compound 255.

To a solution of iraw5-((5)-3-fluoropyrrolidin-l-yl)((45,57?)- & (47?,55)-

4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)methanones SLA 11014 (0.450 g, 1.71 mmol) in methanol (40 mL) was added HCI 37 % (5 mL). After heating at 50 °C for 3 h, the mixture was concentrated and the crude product was coevaporated twice with EtOAc. Trituration with EtOAc and filtration yielded, after drying, to (25,37?)- & (27?,35)-2-amino-l-((5)-3-fluoropyrrolidin-l-yl)-3WO 2006/081273

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212 mg, diastereoisomeric mixture in ratio about 1:1, 97 % yield) as a yellow solid.

threo

Compound 255

MW: 326.19; Yield: 97 %; Yellow Solid; Mp (°C): 168.9

Ή-NMR (CD₃OD, δ): 1.85-2.40 (m, 2H, CH₂), 3.45-4.20 (m, 4H, 2xCH₂),

4.40-4.75 (m, 1H, N-CH), 5.30-5.60 (m, 2H, O-CH & CHF), 8.15-8.25 (m, 1H, ArH), 8.70-8.80 (m, 1H, ArH), 8.90-9.10 (m, 2H, ArH).

MS-ESI m/z (% rel. Int.): 254.1 ([MH]⁺, 81.38), 236.2 (25).

(ffl-S-fluoropyrrolidin- l-yl)((45,57?)- & (47?,56)-4,5-dihydro-5-(thiophen-3yl)oxazol-4-yl)methanones SLA 11016.

To a stirred and cooled 0 °C solution of KOH (0.216 mg, 3.85 mmol) in methanol (8 mL) was added l-((6)-3-fluoropyrrolidin-l-yl)-2-isocyanoethanone VIB 01166 (0.600 g, 4.22 mmol) and thiophene-3-carbaldehyde (0.37 mL, 3.85 mmol). The solution was stirred for 20 h at 0 °C. After evaporation under reduced pressure, the residue obtained was partitioned between EtOAc and H₂O. The product was extracted with EtOAc (4x50 mL) and washed brine (25 mL), dried over MgSO4, filtered and evaporated to yield to a product that was purified using chromatography (florisil, gradient EtOAc:MeOH = 95:5 to 85:15). ((6)-3-fhioropyrrolidin-1^1)((45,57?)- and (47?,56)-4,5-dihydro-5(thiophen-3-yl)oxazol-4-yl)methanones SLA 11016 were obtained as a yellow solid (0.411 g, diastereoisomeric mixture in ratio about 1:1, 32 % yield).

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trans

trans

SLA 11016

MW: 268.31; Yield: 32 %; Yellow Solid; Mp (°C) = 132.9

Ry 0.35 (EtOAc:MeOH = 80:20).

Ή-NMR (CDC1_3j δ): 1.80-2.45 (m, 2H, CH₂), 3.50-4.10 (m, 3H, CH-N & CH₂),

4.20-4.70 (m, 2H, CH₂), 5.15-5.45 (m, 1H, CHF), 6.18-6.25 (m, 1H, O-CH),

6.99 (d, 1H, J= 2.2 Hz, N=CH), 7.00-7.15 (m, 1H, ArH), 7.28-7.35 (m, 1H, ArH), 7.32-7.40 (m, 1H, ArH).

MS-ESI m/z (% rel. Int.): 269.0 ([MH]⁺, 10).

(2£37?)- & (27?,3>S)-2-Amino-l-(6S)-3-fhioropyrrolidin-l-yl)-3-hydroxy-3(thiophen-3-yl)propan-l-one dihydrochlorides Compound 256.

To a solution of ((5)-3-fluoropyrrolidin-l-yl)((45,57?)- & (47?,56)-4,5dihydro-5-(thiophen-3-yl)oxazol-4-yl)methanones SLA 11016 (0.400 g, 1.49 mmol) in methanol (50 mL) was added hydrochloric acid 37 % (4 mL). After heating at 50 °C for 3 h, the mixture was concentrated and the crude product was co-evaporated twice with EtOAc. Trituration with EtOAc and filtration and drying afforded (26',37?)- & (27?,36)-2-amino-l-((6)-3-fluoropyrrolidin-l-yl)-3hydroxy-3-(thiophen-3-yl)propan-l-one dihydrochlorides Compound 256 (451 mg, diastereoisomeric mixture about 1:1,91% yield) as a yellow solid.

Compound 256

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MW: 331.23; Yield: 91 %; Yellow Solid; Mp (°C): 221.6 Ή-NMR (CD3OD, δ): 1.25-2.05 (m, 2H, CH₂), 2.10-2.50 (m, 1H, 0.5xCH₂), 3.20-3.65 (m, 3H, 1.5xCH₂), 3.90-4.10 (m, 1H, CH-N), 4.70-5.10 (m, 2H, O-CH & CHF), 6.92-6.99 (m, 1H, ArH), 7.21-7.32 (m, 2H, ArH). MS-ESI m/z (% rel. Int.): 259.1 ([MH]⁺, 25).

Throughout this specification and the claims which follow, unless the context requires otherwise, the word comprise, and variations such as comprises or comprising, will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

The reference in this specification to any prior publication (or information derived from it), or to any matter which is known, is not, and should not be taken as an acknowledgment or admission or any form of suggestion that that prior publication (or information derived from it) or known matter forms part of the common general knowledge in the field of endeavour to which this specification relates.

Claims

1. A compound represented by the following Formula 1: or a pharmaceutically acceptable salt thereof, wherein:

each R3 is independently selected from H, Ci-₂o alkyl, C3-6 cycloalkyl, aryl-C 1-4 alkyl, aryl(hydroxy)C 1-4 alkyl, COR7 and CO₂R7, the aryl groups optionally being substituted with 13 groups independently selected from halogen, Ci-s alkyl, C1-8 alkoxy and C1-3 thioxy;

R4 is H or CORs;

Rs is Ci-6 alkyl;

s is zero, except when X is N, in which case s is 0 or 1;

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R5 and R6 are independently selected from H, halogen and Ci-6 alkyl, or R5 and R6, together with the atoms to which they are attached, jointly form a carbocyclic or heterocyclic ring having 5 or 6 atoms in the ring, 1 to 3 atoms in the heterocyclic ring being independently selected from N, O and S;

with the proviso that Formula 1 does not cover compounds where R4 is H, Ri and R₂jointly with the nitrogen atom form a pyrrolidine or morpholine ring, the sum of m, n and q is 3, and none of W, X and Y represents a heteroatom.

2. A compound according to Claim 1, wherein the sum of the integers m, n and q is 2 and one or two of W, X and Y is a heteroatom.

3. A compound according to Claim 1, wherein R4 is H.

4. A compound according to Claim 3, wherein both R₃'s are H, or one R3 is H and the other R₃ is COR7 or C1-20 alkyl.

5. A compound according to Claim 1, which is a compound represented by the following formula: or a pharmaceutically acceptable salt thereof, wherein:

Ri, R₂, R4, R5 andR6 are as defined in Claim 1;

each R₃ is independently H, COR7 or CO2R7;

R7 is as defined in Claim 1; and the wavy lines represent bonds of the alpha or beta configuration.

6. A compound according to Claim 1, which is a compound represented by the following formula:

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N' ¹

I D or a pharmaceutically acceptable salt thereof, wherein:

Ri, R₂, R4, Rs andR6 and the wavy lines are as defined in Claim 1; and the R3 S are as defined in Claim 5.

7. A compound according to Claim 1, which is a compound represented by the following formula:

or a pharmaceutically acceptable salt thereof, wherein:

X is O or S;

Ri, R₂, R4, Rs andR6 and the wavy lines are as defined in Claim 1; the R3 S are as defined in Claim 5.

8.

A compound represented by the following Formula 2: or a pharmaceutically acceptable salt thereof, wherein:

Ri and R₂ form a saturated or unsaturated 5- or 6-membered ring together with the nitrogen atom, the ring optionally including one or two additional heteroatoms independently selected from N, O and S, the ring optionally being substituted with one or two COOH,

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CH2OH, OH, B(OH)2, halogen, cyano or C1-6 alkyl groups/atoms, or one or two carbon atoms of the ring being bonded to an oxygen atom to form a keto group, and the ring optionally being condensed with an aromatic or non-aromatic 5- or 6-membered ring which optionally includes one or two heteroatoms selected from N, O and S; each R3 is H, or one R3 is H and the other R3 is benzyl, monohalogen-, dihalogen-, methyl- or methoxysubstituted benzyl, cyclohexyl, C1-7 alkyl, COR7 or COOR7;

R13 and Rio are one of the following combinations of (1) to (3): (1) R13 is NOR11 and Rio does not exist; (2) R13 is OR11 and Rio is C1-6 alkyl; and (3) when the dashed line between carbons 2 and 3 of the propionic acid moiety represents a bond then R13 is C1-6 alkyl; and Rio does not exist;

R11 is H, C1-6 alkyl or COR12;

R12 is C1-6 alkyl;

Rs and R6 are independently selected from H, halogen, C1-6 alkyl, halogen-substituted C1-6 alkyl, C1-6 alkoxy, C1-3 thioxy and phenyl, or R5 and R6, together with the atoms to which they are attached, jointly form a carbocyclic or heterocyclic ring having 5 or 6 atoms in the ring, 1 to 3 of the atoms in the heterocyclic ring being heteroatoms independently selected from N, O and S, and the carbocyclic or heterocyclic ring being optionally substituted with 1 to 6 R9 groups; and each R9 is independently selected from halogen, C1-6 alkyl, C1-6 alkoxy and C1-3 thioxy.

9. A compound according to Claim 8, wherein the sum of the integers m, n and q is 3.

10.

A compound according to Claim 8, wherein one of W, X and Y is N.

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11. A compound according to Claim 8, wherein Ri and R₂ together with the nitrogen to which they are attached form a 5-membered ring.

12. A compound according to Claim 8, wherein R13 is NOR11 and Rio does not exist.

13. A compound according to Claim 8, wherein R13 is NOR11 and Rio is methyl.

14. A compound according to Claim 8, wherein the dashed line between carbons 2 and 3 represents a bond.

15. A compound represented by the following Formula 2: or a pharmaceutically acceptable salt thereof, wherein:

Ri, R₂, R3, Rs, Re, m, n, q, s, W, X, Y, the wavy lines and the dashed lines are as defined in Claim 1;

R13 is NOR11 and Rio does not exist, or R13 is OR11 and Rio is C1-6 alkyl;

R11 is H or COR12; and

R12 is Ci-6 alkyl.

16. A compound according to Claim 8 or Claim 15, wherein the sum of the integers m, n and q is 3.

17. A compound according to Claim 16, wherein one of W, X and Y is N.

18. A pharmaceutical composition comprising a compound as claimed in any one of Claims 1 to 17, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients.

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19. Use of a compound as claimed in any one of Claims 1 to 17, a pharmaceutically acceptable salt thereof, or a composition according to Claim 18, in the manufacture of a medicament for the treatment of pain in a mammal.

20. Use of a compound as claimed in any one of Claims 1 to 17, a pharmaceutically acceptable salt thereof, or a composition according to Claim 18, in the manufacture of a medicament for stimulating the immune system of a mammal.

21. Use of a compound of the formula:

each R3 is independently H, COR7 or CO2R7;

R? is H, Ci -20 alkyl, C3-6 cycloalkyl, aryl, heteroaryl, aryl-Ci-4 alkyl, aryl-(hydroxy)Ci-4 alkyl, heteroaryl-Ci-4 alkyl or heteroaryl-(hydroxy)Ci-4 alkyl, the aryl and heteroaryl groups being optionally substituted with 1 to 3 groups independently selected from halogen, C1-6 alkyl, C1-6 alkoxy and C1-3 thioxy;

R4 is H, Ci-6 alkyl or CORs;

Rs is Ci-6 alkyl;

Rs and Re, are independently selected from H, halogen, C1-6 alkyl, C1-6 alkoxy and C1-3 thioxy, or R5 and R6 together with the atoms to which they are attached jointly form a carbocyclic or heterocyclic ring having 5 or 6 atoms in the ring, the heterocyclic ring having 1 to 3 heteroatoms independently selected from N, O and S, and

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221 the carbocyclic and heterocyclic rings being optionally substituted with 1 to 6 R9 groups; each R9 is independently selected from halogen, C1-6 alkyl and C1-6 alkoxy; and the wavy lines represent bonds of the alpha or beta configuration.

22. Use according to Claim 21, wherein the compound is represented by the following formula:

OR₄ 0 or a pharmaceutically acceptable salt thereof, wherein:

Ri and R2 together with the nitrogen atom to which they are attached form a saturated 5- or 6-membered ring, the ring optionally being substituted with one or two of CO2H, CH2OH, OH, B(OH)2, halogen, cyano or C1-6 alkyl groups;

R3 is as defined in Claim 21; and

R4 is H or C1-6 alkyl.

23. Use according to Claim 22, wherein the compound is represented by the following formula:

OH O

NH₂

2 HCI (‘)'threQ or a pharmaceutically acceptable salt thereof.

24. A method for treating pain in a mammal in need thereof, the method comprising administering to the mammal a compound as claimed in any one of Claims 1 to 17, or a pharmaceutically acceptable salt thereof, or a composition as claimed in Claim 18.

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25. A method for stimulating the immune system of a mammal in need thereof, the method comprising administering to the mammal a compound as claimed in any one of Claims 1 to 17, or a pharmaceutically acceptable salt thereof, or a composition as claimed in Claim 18.

26. A method for treating chronic pain associated with peripheral neuropathy in a mammal in need thereof, the method comprising administering to the mammal a compound as defined in any one of Claims 21 to 23, or a pharmaceutically acceptable salt thereof.