JPH0323536B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention is based on the general formula [In the formula, A is a hydrogen atom or a hydroxyl group; X ¹ is an oxygen atom and X ² is

[Formula] (Z represents a lower alkoxy group) or X ¹ is

In the group represented by [Formula] (Z represents a hydroxymethyl group), X ² represents an oxygen atom or X ¹ represents a

[Formula] (Z is a hydrogen ^atom )

[Formula] (Z represents an amino group which may be protected with an alkoxycarbonyl group or an aralkyloxycarbonyl group);
Alternatively, X ¹ and X ² together represent an oxygen atom, and Y represents a lower alkyl group which may have a hydroxyl group, an amino group which may be protected with a hydroxyl group or an alkoxycarbonyl group. ]However, A is a hydroxyl group,
X ¹

In the formula, when X ² is -CH- (where Z represents a hydroxyl group or an amino group), Y is not a methyl group. ] Pseudo amino sugar [pseudo
−aminosugar; The Journal of Organic Chemistry (J.Org.Chem.) Volume 31,
1516-1521 (1966)
The present invention relates to N-substituted derivatives of the term "sugar", their preparation and uses. The pseudo amino sugar moiety in the above N-substituted derivative of the pseudo amino sugar [], that is, the general formula The pseudo-amino sugar represented by the formula [where A has the same meaning as above] is validamine [a compound in which A in the general formula [] is a hydrogen atom; The Journal of Antibiotics (J.
Antibiotics), Vol. 33, pp. 1575-1576 (1980)]
and valiolamine [compounds in which A in the general formula [] is a hydroxyl group;
55907]. It has been reported that pseudo-amino sugars and their N-substituted derivatives have α-glucosidase inhibitory activity.
-mine, the compound represented by formula (a)) is α-
glucosidase inhibitor Possessing harmful activity [The Journal of
Antibiotics (J.Antibiotics) Volume 33,
pp. 1575-1576 (1980), patent application 1975-136500]. In addition, compounds produced by actinomycetes and having α-glucosidase inhibitory activity that contain valienamine as a constituent component include acarbose (acarbose, BAYg5421, naturwissenschafen), Volume 64,
pp. 535-537 (1977), Special Publication No. 54-39474], Trestatin [(trestatin), 23rd Symposium, The
Chemistry of Natural Products;Symposium
papers), pp. 632-639 (October 1980), Japanese Patent Application Publication No. 1973
-163511], adiposin [(adiposin), TAI-
A, B Others, The Japanese Journal
Of Antibiotics (Jap.J.
Antibiotics, Vol. 36, p. 119 (1981); Starch Sci., Vol. 26, pp. 134-144 (1979), Vol. 27, pp. 107-113 (1980) ), JP-A No. 54-106402, 54-106403, 55-64509, 56-
123986, 56-125398], Amylostatin [(amylostatin), Abstracts of the 4th Carbohydrate Symposium, pp. 58-59 (August 1981), JP-A-1972-
123891, 55-71494, 55-157595], oligostatin [(oligostatin), SF-1130X, JP-A-1973-
26398, 56-43294, The Journal of Antibiotics, Volume 34,
Pages 1424-1433 (1981); Oligostatin C uses hydroxy-validamine instead of valienamine, The Journal of Antibiotics,
Vol. 24, pp. 59-63 (1971))], amino sugar compounds (Japanese Patent Application Laid-open No.
92909) are known. and E. Truscheit regarding α-glucosidase inhibitors of microbial origin, including the above-mentioned compounds.
A review by Angewandte Hemy et al.
Chemie, Vol. 93, pp. 738-755 (1981)]. It has also been reported that the compound represented by formula [b] can be obtained by methanolysis of acarbose and oligostatin C [182nd ACS National Meeting
Abstracts paper) MEDI69, August 1981, New York; The Journal of Antibiotics, Volume 34, 1429~
1433 pages (1981); and Japanese Patent Publication No. 57-24397]. However, the α-glucosidase inhibitory activity of these compounds is still not satisfactory. It is represented by the above general formula []. N-substituted derivatives of validamine and variolamine are new compounds, and it is not known that these compounds exhibit α-glucosidase inhibitory activity. The present inventors have discovered that valienamine, validamine,
As a result of research on various pseudo-amino sugars including variolamine and their various N-substituted derivatives, we found that N-substituted derivatives of pseudo-amino sugars represented by the general formula [] have α-glucosidase inhibitory activity. In particular, the N-substituted derivative of the pseudo-amino sugar represented by the general formula [] has stronger α-glucosidase inhibitory activity than the above-mentioned known compounds having α-glucosidase inhibitory activity. The present invention was completed based on the finding that substituted derivatives have strong α-glucosidase inhibitory activity. That is, the present invention relates to 1 a compound represented by the general formula [], 2 a compound represented by the general formula [] and a compound represented by the general formula [In the formula, X ¹ ′ is an oxygen atom and X ² ′ is

[Formula] (Z′ represents a lower alkoxy group ⁾ or

In the group represented by [Formula] (Z' represents a hydroxymethyl group), X ² ' represents an oxygen atom or X ¹ ' represents a hydroxymethyl group.

[Formula] (Z′ is a ^hydrogen atom)

[Formula] (Z' represents an amino group which may be protected with an alkoxycarbonyl group or an aralkyloxycarbonyl group), or X ¹ ',
Together, X ² ' represents an oxygen atom, and Y' represents an amino group protected by a lower alkyl group, a hydroxyl group, or an alkoxycarbonyl group which may have a hydroxyl group. However, A is a hydroxyl group and X ¹ ' is -
CH ₂ − and X ² ′ is

When [Formula] (where Z' represents a protected amino group), Y is not a methyl group. 3. A method for producing a compound represented by the general formula [], which comprises reacting the compound with a cyclic ketone represented by the general formula [], followed by a reduction reaction, and optionally a deprotection reaction. The present invention relates to an α-glucosidase inhibitor containing a compound represented by: Note that when valienamine, validamine, and variolamine are used as common names to name compounds in the specification of this patent, the position numbers of each carbon atom are assigned as follows.

【formula】

【formula】

[Formula] In the above general formulas [] and [], Y and
Examples of the lower alkyl group represented by Y′ which may have a hydroxyl group include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tart-butyl, and the like having 1 to 1 carbon atoms.
The alkyl group in No. 4 is preferably methyl, and the ethyl group is a lower alkyl group having a hydroxyl group, such as hydroxymethyl, 1-hydroxyethyl,
2-hydroxyethyl (D- and L-glycero configuration), 1,2-dihydroxyethyl (D- and L-glycero configuration), etc. are used. Furthermore, the hydroxyl groups of the amino group, hydroxyl group, hydroxymethyl group, and lower alkyl group having a hydroxyl group represented by Y and Z in the general formula [] may be protected in whole or in part, and the hydroxyl group in the general formula [] Preferably protected. The compound represented by the general formula []

Examples of the substituent represented by [Formula] include: Y is a lower alkyl group which may have a hydroxyl group, X ¹ is an oxygen atom,
X ²

[Formula] (Z represents a lower alkoxy group such as methoxy, ethoxy, propoxy, butoxy, etc.), Y is a hydroxyl group, and X ¹ is

[Formula] (Z represents a hydroxymethyl group), X ² is an oxygen atom, Y is a lower alkyl group which may have a hydroxyl group, X ¹ is a lower alkyl group which may have a hydroxyl group,

[Formula] (Z represents a hydrogen atom), X ² is

[Formula] (Z represents an optionally protected amino group), Y is an optionally protected amino group, and X ¹ is

[Formula] (Z represents a hydrogen atom), X ² is

[Formula] (Z is an optionally protected amino group), Y is a lower alkyl group which may have a hydroxyl group, X ¹ and X ² together represent an oxygen atom, etc. It will be done. Among these, preferred specific examples are as follows:

【formula】

【formula】

【formula】

【formula】

【formula】

【formula】

[expression] or

[Formula]. (However, the amino group may be protected). For the cyclic ketone represented by the general formula [], for example, Y' is a lower alkyl group which may have a hydroxyl group, X ¹ ' is an oxygen atom, and X ² ' is

[Formula] (Z′ is, for example, methoxy, ethoxy, propoxy,
(representing a lower alkoxy group such as butoxy), Y′ is a lower alkyl group which may have a hydroxyl group, X ¹ ′ is a lower alkyl group which may have a hydroxyl group,

[Formula] (Z′ represents a hydrogen atom), X ² ′ is

[Formula] (Z′ represents a protected amino group), Y′ is a protected amino group, and X ¹ ′ is

[Formula] (Z′ represents a hydrogen atom), X ² ′ is

[Formula] (Z′ represents a protected amino group), Y′ is a lower alkyl group which may have a hydroxyl group, X ¹ ′ and X ² ′ together represent an oxygen atom things etc. are used. Among these, preferred specific examples are as follows:

【formula】

【formula】

【formula】

[expression] or

【formula】 (However, -NH-R is a protected amino group
, -OR represents a protected hydroxyl group).
Ru. The compounds represented by the above general formulas [] and []
Hydroxyl groups of compounds (alcoholic and anomeric water)
As a protecting group for hydroxyl groups in sugar chemistry,
Protecting groups used as protective groups, e.g.
acetyl, trifluoroacetyl, methoxy
Acetyl, phenoxyacetyl, propyl, iso
Propyl, pivaloyl, benzoyl, p-nitro
Benzoyl, p-phenylbenzoyl, ethoxy
carbonyl, isobutyloxycarbonyl, ben
Zyloxycarbonyl, p-nitrophenoxyca
rubonyl, 3-benzoylpropionyl, benzo
Acyl-type protecting groups such as ylformyl, methyl,
Chyl, propyl, isopropyl, butyl, isobutyl
Chil, sec-butyl, tert-butyl, ethoxye
tyl, allyl, trityl, trimethylsilyl, di
Methylethylsilyl, benzyl, p-methoxybe
Ether-type protecting groups such as glycolide-type protecting groups such as
(including protective groups), methylene, ethylidene, isopropylene
Pylidene, methoxymethylene, ethoxymethylene
methoxyethylidene, dimethoxymethylene,
Cyclopropylidene, cyclopentylidene, cyclopropylidene
lohexylidene, benzylidene, tetrahydropylidene
Ranyl, methoxytetrahydropyranyl, etc.
Setal or ketal protecting groups are used.
Ru. The compounds represented by the above general formulas [] and []
As a protecting group for the amino group of the compound, amino sugar,
Amino in Minocyclitol and Peptide Chemistry
A protecting group used as a protecting group for a group, e.g.
Lumyl, acetyl, propionyl, butyryl, etc.
alkanoyl group, benzoyl, p-chloroben
Aroyl such as zoyl and p-nitrobenzoyl
group, methoxycarbonyl, ethoxycarbonyl,
1-propoxycarbonyl, tert-butoxycal
Alkoxycarbonyl groups such as Bonyl, benzyl
Aralkyloxycarbo such as oxycarbonyl
In addition to acyl-type protecting groups such as nyl groups, 2,4-di
A nitrophenyl group or the like is used. N- of the pseudo-amino sugar represented by the general formula []
Specific examples of substituted derivatives include (1) Methyl 4-[(1S,2S)-(2,4/3,5)
-2,3,4-trihydroxy-5-hydroxy
dimethylcyclohexyl]amino-4,6-di
Deoxy-α-D-glucopyranoside (2) Methyl 4-[(1S,2S)-(2,4/3,5)
-2,3,4-trihydroxy-5-hydroxy
dimethylcyclohexyl]amino-4,6-di
Deoxy-α-D-galactopyranoside (3) Methyl 4-[(1S,2S)-(2,4,5
(OH)/3,5(CH₂OH))-2,3,4,5
-tetrahydroxy-5-hydroxymethylsilyl
Chlohexyl]amino-4,6-dideoxy-
α-D-glucopyranoside (4) Methyl 4-[(1S,2S)-(2,4,5
(OH)/3,5(CH₂OH))-2,3,4,5
Tetrahydroxy-5-hydroxymethylsic
lohexyl]amino-4,6-dideoxy-α
-D-galactopyranoside (5) Methyl 4-[(1S,2S)-(2,4/3,5)
-2,3,4-trihydroxy-5-hydroxy
dimethylcyclohexyl]amino-4-deoxy
C-α-D-glucopyranoside (6) Methyl 4-[(1S,2S)-(2,4/3,5)
-2,3,4-trihydroxy-5-hydroxy
dimethylcyclohexyl]amino-4-deoxy
C-α-D-galactopyranoside (7) Methyl 4-[(1S,2S)-(2,4,5
(OH)/3,5(CH₂OH))-2,3,4,5
-tetrahydroxy-5-hydroxymethylsilyl
Chlohexyl]amino-4-deoxy-α-D
-Glucopyranoside (8) Methyl 4-[(1S,2S)-(2,4,5
(OH)/3,5(CH₂OH))-2,3,4,5
-tetrahydroxy-5-hydroxymethylsilyl
Chlohexyl]amino-4-deoxy-α-D
-galactopyranoside (9) [(1S, 2S) - (2, 4/3, 5) - 2, 3,
4-trihydroxy-5-hydroxymethylsilyl
Chlorhexyl] [(2S)-(2,6/3,4)-4
-benzyloxycarbonylamino-2,3-
dihydroxy-6-hydroxymethylcyclohe
xyl amine (10) [(1S, 2S) - (2, 4/3, 5) - 2, 3,
4-trihydroxy-5-hydroxymethylsilyl
Chlorhexyl] [(1R,2S)-(2,6/3,4)
-4-amino-2,3-dihydroxy-6-hydroxy
Droxymethylcyclohexylamine (11) [(1S, 2S) - (2, 4/3, 5) - 2, 3,
4-trihydroxy-5-hydroxymethylsilyl
Chlorhexyl] [(1S, 2S) - (2, 6/3, 4)
-4-amino-2,3-dihydroxy-6-hydroxy
Droxymethylcyclohexylamine (12) [(1S, 2S) - (2, 4, 5 (OH) / 3, 5
(CH₂OH))-2,3,4,5-tetrahydro
xy-5-hydroxymethylcyclohexyl]
[(1R,2S)-(2,6/3,4)-4-benzyl
Oxycarbonylamino-2,3-dihydroxy
C-6-hydroxymethylcyclohexyl]a
min (13) [(1S, 2S) - (2,4,5(OH)/3,5
(CH₂OH))-2,3,4,5-tetrahydro
xy-5-hydroxymethylcyclohexyl]
[(1S,2S)-(2,6/3,4)-4-benzyl
Oxycarbonylamino-2,3-dihydroxy
C-6-hydroxymethylcyclohexyl]a
min (14) [(1S, 2S) - (2,4,5(OH)/3,5
(CH₂OH))-2,3,4,5-tetrahydro
xy-5-hydroxymethylcyclohexyl]
[(1R,2S)-(2,6/3,4)-4-amino-
2,3-dihydroxy-6-hydroxymethyl
cyclohexyl amine (15) [(1S, 2S) - (2,4,5(OH)/3,5
(CH₂OH))-2,3,4,5-tetrahydro
xy-5-hydroxymethylcyclohexyl]
[(1S,2S)-(2,6/3,4)-4-amino-
2,3-dihydroxy-6-hydroxymethyl
cyclohexyl amine (16) N-(2,3,5-trihydroxy-6-
Hydroxymethyltetrahydropyran-4-y
) Variolamine (17) N-(2,3-O-isopropylidene-2,
3-dihydroxy-5-(1,2-dihydroxy
ethyl)tetrahydrofuran-4-yl]b
Liolamine (18) N-(2,4-diethoxycarbonylamine
(5,6-dihydroxycyclohexyl)b
Liolamine (19) N-(2,4-diamino-5,6-dihyde
Roxycyclohexyl)variolamine (20) N-(2,4-diethoxycarbonylamine
(5,6-dihydroxycyclohexyl)b
Ridamine (21) N-(2,4-diamino-5,6-dihyde
roxycyclohexyl)validamine (22) Methyl 4-[(1S,6S)-(4,6/5)-
4,5,6-trihydroxy-3-hydroxy
Methyl-2-cyclohexen-1-yl]amide
No-4,6-dideoxy-α-D-galactopi
Ranocide (23) N-(2,3,5-trihydroxy-6-
Hydroxymethyltetrahydropyran-4-y
) Valienamine (24) N-(2,3,O-isopropylidene-2,
3-dihydroxy-5-(1,2-dihydroxy
ethyl)tetrahydrofuran-4-yl]b
Lienamine (25) Methyl 4-[(1S,6S)-(4,6/5)-
4,5,6-trihydroxy-3-hydroxy
Methyl-2-cyclohexen-1-yl]amide
No-4-dideoxy-α-D-glucopyranosyl
de (26) Methyl 4-[(1S,6S)-(4,6/5)-
4,5,6-trihydroxy-3-hydroxy
Methyl-2-cyclohexen-1-yl]amide
No-4-deoxy-α-D-galactopyranosi
de Examples include. The α-glucosidase inhibitor of the present invention can be used in humans and
inhibits carbohydrate metabolism in humans and non-human animals
Therefore, for example, it has a blood sugar rise suppressing effect,
Hyperglycemic symptoms and various diseases caused by hyperglycemia,
For example, obesity, adiposity, hyperlipidemia (arterial
sclerosis), diabetes, prediabetes, and oral microbial
Prevention of diseases caused by sugar metabolism, such as tooth decay
It is a compound useful for Also, pseudo amino sugar N-
Foods manufactured by adding substituted derivatives []
As a food for patients with metabolic disorders, and as a food for preventing metabolic disorders.
It is also suitable for healthy people. Also, low fat
Feed additives for livestock to obtain quality edible meat.
It is also useful. Therefore, the α-glue of the present invention
Cosidase inhibitors are used in pharmaceuticals and food additives,
It is useful as a food additive. α of the present invention
-Glucosidase inhibitors can be administered orally or parenterally
, preferably orally. The above N-substituted derivatives of pseudo-amino sugars []
Stable crystals or powder with almost no toxicity (labeled
Tsuto LD₅₀500mg/Kg or more), free base or hydrated
It can be used as a drug and can be used as a drug by normal methods.
Any non-toxic acid addition salt with a chemically acceptable acid
It can also be used as As such acid
For example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid,
Inorganic acids such as nitric acid, acetic acid, malic acid, citric acid,
Ascorbic acid, mandelic acid, methanesulfonic acid
Organic acids such as are used. Vario like this
amine and validamine derivatives alone or
Use by mixing with a non-toxic carrier. For example, coffee,
Soft drinks, fruit juice, beer, milk, diam, a
liquid or solid foods such as jelly, seasonings,
Alternatively, it can be used with various staple foods and side dishes.
It can be used directly or in the form of food additives.
or can be taken before or after meals.
can. Furthermore, you can obtain low-fat, high-quality edible meat.
It can also be used as a feed additive for livestock.
Ru. The α-glucosidase inhibitor of the present invention includes, for example,
For example, water, ethanol, ethylene glycol, poly
Liquid carriers such as ethylene glycol, starch,
Non-toxic solid carriers such as lulose and polyamide powder
Dilute with a carrier to prepare ampoules, granules, tablets,
Although it is commonly used for pills, capsules, syrups, etc.
can be prepared and used for the various purposes mentioned above.
Ru. Sweeteners, preservatives, dispersants, and colorants are also common.
can be used. Specifically, for example, the compound [] about 10 to 400
By taking tablets containing mg after every meal,
Suppresses the increase in blood glucose concentration caused by eating
can do. Also, for example, carbohydrates in food
About 0.0001 to 1% of the content of compounds []
It may be added to various foods. When mixed with feed, check the carbohydrate content of the feed.
0.0001 to 1% is desirable. N-substitution induction of pseudo-amino sugars included in the present invention
Both compounds [] are new compounds that have not been described in the literature.
For example, it can be synthesized by the following method.
I can do it. That is, in a suitable solvent, validamate
Pseudo amino sugars such as amine, variolamine, etc.
, alcoholic hydroxyl group and hydro
The oxymethyl group may be protected and the anomeric
(i.e. glycosidic) hydroxyl group is
protected aldoketoses; or protected
Optionally protected hydroxyl group, protected
hydroxymethyl group, which may be
It has two or more of the following amino groups:
obtained by reacting cycloalkanones with
The Schiff base is subjected to a reduction reaction and protected if necessary.
It can be synthesized by subjecting it to a group elimination reaction.
can. Amino group of pseudo amino sugar [] and the above
Compound [], i.e. aldoketoses or scissors
Condensation reaction of clohexanone with carbonyl group,
The subsequent reduction reaction and protective group elimination reaction are
It may be carried out continuously in the same reaction vessel, or
It can be done in two stages, each separately.
It may be done in three stages. Between the pseudo amino sugar [] and the above compound []
Reactions in condensation reactions and subsequent reduction reactions
Examples of solvents include water, methanol, and propane.
Alcohols such as alcohol and butanol, dimethyl
Sulfoxide, dimethylformamide, N-methy
Ruacetamide, methylcellosolve, dimethylcetamide
Rosolve, diethylene glycol dimethyl ether
glyme such as alcohol, dioxane, tetrahydrofu
Polar solvents such as ran, acetonitrile, or
These mixed solvents or those polar solvents and
With non-polar solvents such as loloform and dichloromethane
Mixtures can be used. The reaction temperature in the Schiff base formation reaction is
Although not limited to, usually at room temperature or around 100℃
This is done by heating up to. Reaction time depends on reaction temperature
and the type of compound used.
Although there are differences, the reaction usually takes a few minutes to 24 hours.
By doing so, you can achieve your goals. For the reduction reaction of the Schiff base formed, each
Species of metal hydride complex reducing agents, e.g. borohydrides
Sodium borohydride, potassium borohydride, borohydride
Lithium uride, sodium trimethoxyborohydride
Alkali metal borohydrides such as um, cyanohydrogen
Cyanoborohydride such as sodium borohydride
Hydrogen such as potash metal, lithium aluminum hydride, etc.
aluminum alkali metal, dimethylamine boron
Dialkylamine borane such as oran are advantageously used.
It will be done. In addition, cyanoalkali metal hydride, e.g.
When using sodium cyanoborohydride
is in the presence of acidic conditions, e.g. hydrochloric acid acetic acid etc.
Preferably, the reaction is carried out. The reaction temperature is not particularly limited, but is usually room temperature,
In some cases, especially at the beginning of the reaction, ice cooling may be used.
and in some cases heated up to around 100℃
Schiff base and reducing agent
There are differences depending on the type. Reaction time and reaction temperature
Depending on the type of Schiff base and reducing agent being reduced,
It varies depending on the situation, but it usually takes a few minutes to 24 hours.
You can achieve your goal by reacting
Wear. In the reduction reaction of the Schiff base formed
Therefore, the pseudo amino sugar [ ] used as a raw material was
Unsaturated like liolamine and validamine
Means of catalytic reduction if there are no double bonds
You can also use That is, the Schiff base is
hydrogen gas in the presence of a catalyst for catalytic reduction in a suitable solvent.
This is done by shaking or stirring in a flowing stream.
Examples of catalysts for catalytic reduction include platinum black, diacid
Platinum chloride, palladium black, palladium carbon, la
Commonly used solvents include Nönickel, etc.
For example, water, methanol, ethanol
Alcohols such as dioxane, tetrahydrofu
Ran, dimethylformamide or a mixture thereof.
A mixed solvent or the like is used. The reaction is usually carried out at room temperature and normal pressure.
However, it may also be carried out under pressure.
You can warm it up. Removal of hydroxyl-protecting group and amino-protecting group
The reaction can be carried out using methods known per se.
can. For example, O-cyclohexylidene group, O
-isopropylidene group, O-benzylidene group, O
- Trityl group etc. are hydrochloric acid, acetic acid, sulfonic acid type
By hydrolyzing with acids such as on-exchange resins,
, O-acetyl group, N-ethoxycarbonyl
group, N-benzyloxycarbonyl group, N-phthalate
Lyle groups, etc. can be used in ammonia water, barium hydroxide, hydrates, etc.
Hydrolysis with an alkali such as water hydrazine
also N-benzyloxycarbonyl
The group can also be eliminated by hydrogenolysis with catalytic reduction.
can be done. In addition, it is used as a raw material in the above reaction.
Water of pseudo amino sugar represented by the general formula [ ]
The acid group is a hydroxyl group protecting group (for example, the general formula [] or
The hydroxyl group in the compound represented by and []
Protected with the above-mentioned protecting group)
Good too. Expressed by the general formula [] obtained in this way,
The compounds and their synthetic intermediates are self-contained.
by means known in the art, such as filtration, centrifugation, concentration, reduction.
Pressure concentration, drying, freeze drying, adsorption, desorption, various solvents
A method that utilizes the difference in solubility in
solvent extraction, dissolution, precipitation, crystallization, recrystallization, etc.)
Chromatography (e.g. ion exchange resin,
Activated carbon, high porous polymer, safety adhesive
S, Cephadex ion exchanger, Cellulose,
Ion exchange cellulose, silica gel, alumina
chromatography), etc.
Can be separated and purified. Variol is one of the raw material compounds used in this application.
Ruamine (in the general formula [], A is represented by a hydroxyl group)
compounds) are described in, for example, Japanese Patent Application No. 56-55907.
Cultivating microorganisms belonging to the genus Streptomyces
By the method of patent application No. 56-64370 and
There is valienamine described in the patent application No. 56-144309.
An organic chemical synthesis method using validamine as a raw material
It can be manufactured by. In addition, the compound represented by the general formula []
For example, by the methods described in Figures 1, 2 and 3.
can be manufactured. In addition, valienamine is used instead of the raw material compound [ ].
The bar corresponding to the compound [] synthesized using
N-substituted derivatives of lienamine also have strong α-glucose
It has anti-dase activity. (In the above formula, Ac is acetyl, Ph is phenyl,
Cbz is benzyloxycarbonyl, DMSO is dimethyl
Indicates tylsulfoxide. ) Test examples, reference examples, and working examples are listed below.
The content of the invention will be described in detail, but the scope of the invention will not be limited to these details.
It is not limited. Test example Method for measuring glucosidase inhibitory activity When using maltose and sucrose as substrates,
Maltase prepared from the small intestine mucosa of pigs
Bisatsucalase [Borgström (B.
Borgstrom) and Dahlquist (A.
Acta Chemica Scandi by Dahlqvist
Navica (Acta Chem.Scand.) Volume 12, 1997-2006
[prepared according to the method described in P., 1958]
Inhibitory activity was determined in 0.02M phosphate buffer solution (PH6.8).
Inhibition to be tested in 0.25 ml of appropriately diluted enzyme solution
0.5ml of the same buffer solution of the substance and 0.05M of the substrate
0.25ml of tose or 0.05M sucrose in the same buffer solution
was added and the mixture was allowed to react at 37°C for 10 minutes.
Lucose B-test reagent (glucose for dextrose measurement)
Add 3 ml of oxidase reagent (manufactured by Wako Pure Chemical Industries).
Then, further heat at 37℃ for 20 minutes, and adjust the wavelength of the reaction solution to 505nm.
It was calculated by measuring the absorbance at . N-substitution induction of pseudo-amino sugars described in Examples
50% inhibition of body maltase (pig, intestinal mucosa)
Concentration (hereinafter referred to as IC)₅₀(abbreviated as maltase)
and 50% inhibition of satucalase (pig, intestinal mucosa).
Harmful concentration (hereinafter referred to as IC)₅₀(abbreviated as Satukalase)]
is 3 to 5 different types for each compound.
Inhibition rate measured using the above assay at the same concentration.
(%). Purification of each compound described in Reference Examples and Examples
Column chromatography elution fraction during the process
Minutes are usually thin layer chromatography (TLC)
Check the ingredients and find a picture that contains the necessary ingredients.
A portion was collected and used for the next step. Described in Examples
The TLC Rf value of each compound is not particularly specified.
As long as the thin plate is pre-coated (Pre-
coated) TLC plate silica gel 60F₂₅₄(Mel
n-
Propyl alcohol, acetic acid, water (4:1:1)
It was measured using (as a control sample in the above method)
Measured Rf value of pseudo amino sugar: Valienamine Rf
= 0.42, Validamine Rf = 0.35, Variolamine
Rf=0.30) The symbols used in reference examples and working examples are as follows.
It has great significance. s, singlet; d, doublet; dd, da
double doublet; t, triplet; q, medical record
m, multiplet; J, coupling constant;
DMSO, dimethyl sulfoxide; DMF, dimethylene
Ruformamide Reference example 1 N-benzyloxycarbonylvalienamine Valienamine (100g) and sodium bicarbonate
Dissolve um (100g) in water (1) and heat at 5-10℃
benzyloxycarbonyl chloride under cooling to
After dropping a solution of (170ml) in toluene (500ml),
Stir at room temperature for 1 hour and then at room temperature for 3 hours. anti
After cooling the reaction solution to below 10℃ and leaving it for 2 hours, precipitation occurs.
of N-benzyloxycarbonylvalienamine
Filter the crystals, wash with water and toluene, and dry.
Ru. Combine the filtrate and washing liquid, separate the aqueous layer, and add toluene.
After washing with water, adjust the pH to 5 to 5.5 with 2N hydrochloric acid, and
Concentrate to 400ml. Leave the concentrate in the refrigerator overnight
and the precipitated N-benzyloxycarbonyl fluoride
Filter the enamine crystals, wash with cold water, and dry.
Ru. Yield: 146g. [α]^{twenty four} _D+121.7゜(c=1,H₂O) Elemental analysis: C₁₅H₁₉NO.₆ Calculated value (%): C, 58.24; H, 6.19; N, 4.53 Experimental value (%): C, 58.31; H, 6.17; N, 4.49 NMR (CMSO−d₆) δ: 5.02 (2H, s), 5.46
(1H, d, J = 5Hz), 6.67 (1H, d, J
= 8.5Hz), 7.33 (5H, s) Reference example 2 N-tert-butoxycarbonylvalienamine Dissolve valienamine (10g) in water (50ml),
After adding dioxane (50ml), tert-butyl
4,6-dimethylpyrimidin-2-ylthiocal
Add Bonato (16.8g) and stir at room temperature for 18 hours.
Ru. The reaction solution was concentrated under reduced pressure, and the residue was dissolved in water (200ml).
After washing with ethyl acetate, reduce the pressure to about 50ml.
Concentrate and leave in the refrigerator overnight. precipitated crystals
Filter off and wash with cold water. Combine the filtrate and washing liquid
MCI Gel CHP20P (manufactured by Mitsubishi Chemical Industries) (400ml)
After applying column chromatography and washing the column with water,
Elute with % methanol water. Concentrate the eluted fraction under reduced pressure
Amberlight CG-50 (H⁺type, roam a
Column chromatography (manufactured by Haas Inc., USA) (550ml)
eluted with water. Concentrate the eluted fraction under reduced pressure
When the concentrate is lyophilized, N-tert-butoxy
Carbonylvalienamine white powder (12.5g)
can get. [α]^{twenty three} _D+128.2゜(c=1,H₂O) Elemental analysis: C₁₂H_{twenty one}NO.₆ Calculated value (%): C, 52.35; H, 7.69; N, 5.09 Experimental value (%): C, 52.09; H, 7.93; N, 5.02 NMR(D₂O) δ: 1.58 (9H, s) Reference example 3 9-bromo-6,7,8-trihydroxy-1
-Hydroxymethyl-3-oxo-2-oxa-
4-Azabicyclo [3.3.1] Nonane N-benzyloxycarbonylvalienamine
(146g) in methanol (1.2),
Bromine (75 g) was added to methanol (450 g) while cooling to 5°C.
ml) solution dropwise. Stir the reaction solution at the same temperature for 1 hour.
After stirring, concentrate under reduced pressure. Ethanol-acetic acid to the residue
Add ethyl (1:10) (2.2) and refrigerate overnight.
The precipitated crystals were collected by filtration and diluted with ethyl acetate or
After washing with oil and petroleum ether and drying, 9-bro
Mo-6,7,8-trihydroxy-1-hydroxy
dimethyl-3-oxo-2-oxa-4-azabi
Cyclo[3.3.1]nonane (136 g) is obtained. child
This is recrystallized from water. [α]^{twenty four} _D+42.1゜(c=1,H₂O) Elemental analysis: C₈H₁₂NO.₆Br・H₂O Calculated value (%): C, 30.39; H, 4.46; N,
4.43; Br, 25.28 Experimental value (%): C, 30.31; H, 4.52; N,
4.40; Br, 25.38 IR ν^KBr _naxcm^-1:1700 (C=0) NMR(D₂O) δ: 3.66 (1H, t, J = 9.3Hz),
3.93 (1H, t, J = 3.2Hz), 3.95 (1H, d,
J = 13Hz), 4.14 (1H, d, J = 13Hz),
4.18 (1H, d, J = 9.3Hz), 4.34 (1H, dd,
J=3Hz, 9.3Hz), ~4.75 (1H) Reference example 4 9-bromo-6,7,8-trihydroxy-1
-Hydroxymethyl-3-oxo-2-oxa-
4-Azabicyclo [3.3.1] Nonane N-tert-butoxycarbonylvalienamine
Dissolve (2.8g) in methanol (30ml) and add to ice water.
Dissolve bromine (1.8 g) in methanol (200 ml) under cooling.
Add the liquid dropwise and stir for 1 hour. Concentrate the reaction solution under reduced pressure
and remove the residue with MCI gel CHP20P (manufactured by Mitsubishi Chemical Industries, Ltd.,
250ml) column chromatography and elute with water.
The eluted fraction is concentrated under reduced pressure and the concentrated solution is lyophilized.
A white powder (2.5 g) is obtained. This white powder
(40 ml of acetic acid), heated at 80 to 90°C for 30 minutes, and then
When left at room temperature overnight, 9-bromo-6,7,8-
rihydroxy-1-hydroxymethyl-3-oxy
So-2-oxa-4-azabicyclo [3.3.1]
Naan crystals (2.1 g are obtained. Elemental analysis: C₈H₁₂NO.₆Br Calculated value (%): C, 32.23; H, 4.06; N,
4.70; Br, 26.81 Experimental value (%): C, 32.11; H, 4.19; N,
4.87; Br, 26.67 Reference example 5 6,7,8-trihydroxy-1-hydroxy
Methyl-3-oxo-2-oxa-4-azabisi
Black [3.3.1] Nonane 9-bromo-6,7,8-trihydroxy-1
-Hydroxymethyl-3-oxo-2-oxa-
4-Azabicyclo [3.3.1] Nonane (100g) in water
(1.3) and hydrogen while keeping at 25-30℃.
Add sodium boronate (55g) little by little.
After stirring the reaction solution at the same temperature for 2 hours, acetic acid was added to adjust the pH.
Adjust to 6-7 and concentrate under reduced pressure to about 800 ml. concentrated
Subject the liquid to activated carbon (4.8) column chromatography,
After washing the column with water (5), dissolve it in 50% methanol water.
put out The eluted fractions were concentrated under reduced pressure, and the residue was
Add water (350 ml) and heat under reflux for 10 to 20 minutes.
Leave it in the refrigerator overnight, filter out the precipitated crystals,
After washing with cold methanol and drying, 6,7,8-
trihydroxy-1-hydroxymethyl-3-o
Xo-2-oxa-4-azabicyclo [3.3.1]
Nonane (64 g) is obtained. [α]^{twenty four} _D+35.0゜(c=1,H₂O) Elemental analysis: C₈H₁₃NO.₆ Calculated value (%): C, 43.83; H, 5.98; N, 6.39 Experimental value (%): C, 43.80; H, 5.96; N, 6.52 IR ν^KBr _naxcm^-1:1670 (C=0) NMR(D₂O) δ: 2.07 (1H, dd, J=2Hz, 15
Hz), 2.34 (1H, dd, J=5Hz, 15Hz),
3.45~4.1 (6H) mp 254-255℃ (decomposition) Reference example 6 9-iodo-1-hydroxymethyl-6,7,
8-trihydroxy-3-oxo-2-oxa-
4-Azabicyclo [3.3.1] Nonane a N-benzyloxycarbonylvalienamine
50ml of 3.1g methanol solution and 2.6g iodine
20ml of methanol solution was cooled to 5-10℃.
Drop simultaneously into 20 ml of tanol. Pour the reaction solution into the chamber.
After stirring at room temperature for 25 hours, it was concentrated under reduced pressure and the residue was dissolved in acetic acid.
After adding to the ethyl and water mixture and distributing it, the water
Separate the layers. After washing the aqueous layer with ethyl acetate, reduce
Concentrate under pressure and transfer the residue to MCI gel CHP20P (250ml,
(manufactured by Mitsubishi Chemical Industries) column chromatography,
Elutes with After concentrating the eluate fraction under reduced pressure, freeze-dry it.
Then, 9-iodo-1-hydroxymethyl-
6,7,8-trihydroxy-3-oxo-2
-oxa-4-azabicyclo[3.3.1]nonane
1.1g is obtained. b N-benzyloxycal cooled to 0-5°C
Bonylvalienamine 3.1g methanol solution 50
1.7 g of iodine monochloride in acetonitrile solution
Add 20ml dropwise and keep at 0-5℃ for 5 hours, then at room temperature.
Stir for 15 hours. Concentrate the reaction solution under reduced pressure and remove the residue.
was added to a mixture of ethyl acetate and water and partitioned.
After that, separate the aqueous layer. Wash the aqueous layer with ethyl acetate
After that, adjust the pH to 5 with saturated sodium bicarbonate solution.
and concentrate under reduced pressure. MCI gel CHP20P residue
(250ml, manufactured by Mitsubishi Chemical Industries) column chromatography
Attach and elute with water. Concentrate the eluted fraction under reduced pressure,
Dissolve the residue in a small amount of water by heating and store in the refrigerator overnight.
9-iodo-1-hydroxymethylene
Ru-6,7,8-trihydroxy-3-oxo
-2-oxa-4-azabicyclo [3.3.1]
3.1 g of naan crystals are obtained. [α]^{twenty four} _D+37.5° (c=1, H₂O) Elemental analysis: C₈H₁₂NO.₆I・H₂O Calculated value (%): C, 26.46; H, 3.89; N, 3.86 Experimental value (%): C, 26.57; H, 3.97; N, 3.96 IR ν^KBr _naxcm^-1;1680 (C=0) NMR(D₂O) δ: 3.70 (1H, t, J = 9.5Hz),
3.92 (1H, t, J = 3.5Hz), 4.02 and 4.28
(1H, d, J = 15Hz each), 4.33 (1H, d,
J = 9.5Hz), 4.53 (1H, dd, J = 3.5PH, 9
Hz), 4.79 (1H, d, J = 3.5Hz) Reference example 7 1L(1S)-(1(OH),2,4,5/1,3)-5
-amino-1-hydroxymethyl-1,2,3,
4-Cyclohexanetetrol (variolamide)
hmm) 6,7,8-trihydroxy-1-hydroxy
Methyl-3-oxo-2-oxa-4-azabisi
Chlo [3.3.1] Dissolve nonane (50g) in water (1.5)
Dissolve and add barium hydroxide (200g) to 70-80%
Stir for 3 hours while heating to °C. Cool the reaction mixture to 20℃
After cooling, pass carbon dioxide gas through for 30 minutes to remove the resulting precipitate.
Filter off and wash with water. Combine the filtrate and washing liquid and
-Light CG-50 (NH⁺ _Fourtype, loam and ha
Attached to a column chromatograph manufactured by Co., Ltd. (USA), 1.6)
vinegar. After washing the column with water (1), add 0.2N ammonia
Elutes with water. Concentrate the eluted fraction under reduced pressure to approximately 200 ml.
Pour the concentrate into a 1×2 Dowex (OH^-type, dau ke
Column chromatography manufactured by Michal (USA), 1.6)
Attach and elute with water. After concentrating the eluate fraction under reduced pressure, freeze it.
When dried, a white powder of variolamine (42.7
g) is obtained. [α]^{twenty five} _D+19.6゜(c=1,H₂O) Elemental analysis: C₇H₁₅NO._Five・H₂O Calculated value (%): C, 39.80; H, 8.11; N, 6.63 Experimental value (%): C, 39.87; H, 8.13; N, 6.68 NMR(D₂O) δ: 1.80 (1H, dd, J=3.8Hz,
15.5Hz), 2.07 (1H, dd, J=3Hz, 15.5
Hz), 3.4-3.6 (1H), 3.55 (1H, d, J =
10Hz), 3.63 (2H), 3.72 (1H, dd, J=4.2
Hz, 10Hz), 3.99 (1H, t, J = 10Hz) Reference example 8 Methyl 4-O-benzoyl-2,3-O-cy
Chlohexylidene-6-bromo-6-deoxy-
α-D-glucopyranoside Methyl 4-O-benzoyl-6-bromo-6
-deoxy-α-D-glucopyranoside (25g)
Dissolve in dimethylformamide (50ml) and add 1,
1-dimethoxycyclohexane (50ml) and p-t
Add luenesulfonic acid (1.0g) and aspirate
At 55℃ for 5 hours under vacuum of 40mmHg
Stir for a while. The reaction solution was heated at the same temperature under reduced pressure (18 to 20 mm
Hg) for 30 min. The residue was dissolved in ethyl acetate.
Add to water mixture and dispense. Separate the ethyl acetate layer.
taken, washed with saturated sodium bicarbonate solution and sulfurized.
After drying with sodium chloride, concentrate under reduced pressure. residue
Subjected to silica gel (600ml) column chromatography,
After washing the column with toluene, toluene-ethyl acetate
Elute at 9:1. Concentrate the eluted fraction under reduced pressure
After that, dry it under reduced pressure in a desiccator overnight.
Chil 4-O-benzoyl-2,3-O-cyclo
hexylidene-6-bromo-6-deoxy-α-
D-glucopyranoside syrup (26g)
is obtained. Elemental analysis: C₂₀H_{twenty five}O₆Br Calculated value (%): C, 54.43; H, 5.71; Br, 18.11 Experimental value (%): C, 54.99; H, 5.96; Br, 18.56 Reference example 9 Methyl 2,3-O-cyclohexylidene-6
-deoxy-α-D-glucopyranoside Methyl 4-O-benzoyl-2,3-O-cy
Chlohexylidene-6-bromo-6-deoxy-
α-D-glucopyranoside (5.0g)
Dissolve in Dorofuran (50ml) and cool with ice water.
Add lithium aluminum chloride (1.0g) to 30
After stirring for a minute, the mixture is further stirred for 2 hours while heating under reflux.
After cooling the reaction solution again with ice water and adding water, the pressure was reduced.
Concentrate. Add the residue to a mixture of ethyl acetate and water.
Then, while stirring, adjust the pH to below 2 with 2N hydrochloric acid.
Separate the ethyl acetate layer and add saturated sodium hydrogen carbonate.
After washing with solution and drying with sodium sulfate, concentrate under reduced pressure.
Shrink. Transfer the residue to a column of silica gel (250ml)
Chromatographed toluene-ethyl acetate (3:
Elute with 1). After concentrating the eluted fraction under reduced pressure, leave it overnight.
When dried under reduced pressure in a siccator, methyl 2,
3-O-cyclohexylidene-6-deoxy-α
-D-glucopyranoside syrupy substance (2.8
g) is obtained. [α]^{twenty four} _D+88.6° (c=1, CH₃OH) Elemental analysis: C₁₃H_{twenty two}O_Five Calculated value (%): C, 60.44; H, 8.59 Experimental value (%): C, 60.25; H, 8.52 NMR (CDCl)₃) δ: 1.30 (3Hd, J=6Hz),
1.25-1.9 (10H, m), 2.98 (1H, d, J=
3Hz), 3.43 (3H, s), 3.3~3.65 (3H,
m), 3.93 (1H, t, J = 9.5Hz), 4.96
(1H, d, J=3Hz) Reference example 10 Methyl 2,3-O-cyclohexylidene-6
-deoxy-α-D-xylo-hexopyranoside
-4-ulose (methyl 2,3-O-cyclohe
Xylidene-6-deoxy-α-D-xylo-4
-hexopyranose uroside) Trifluoroacetic anhydride (11.1 ml) dichloromethane
(20 ml) solution in dimethyl sulfoxide (7.5
ml) in dichloromethane (20 ml) at -65℃ or below.
while cooling, and stirred at the same temperature for 10 minutes. child
Add methyl 2,3 to the reaction solution while cooling to -70℃ or below.
-O-cyclohexylidene-6-deoxy-α-
D-glucopyranoside (6.8g) in dichloromethane
(30 ml) solution was added dropwise and stirred at the same temperature for 1 hour.
Ru. This reaction solution was cooled to below -65°C with triethyl alcohol.
After adding Ruamine (22.2ml), remove the cooling bath.
Stir until the reaction temperature reaches 20°C. Place the reaction mixture on ice.
Add dichloromethane to the mixture of water and dichloromethane.
Separate the tan layer and add 2N hydrochloric acid and saturated sodium bicarbonate.
Washed with thorium solution and dried with sodium sulfate
Afterwards, the solvent is distilled off. Remove the residue from silica gel (400
ml) column chromatography and toluene-acetic acid ethyl
Elute with chill (17:3). Concentrate the eluted fraction under reduced pressure
and dry the residue under reduced pressure in a desiccator overnight.
Then methyl 2,3-O-cyclohexylidene
-6-deoxy-α-D-xylo-hexopyrano
Crystals of side-4-ulose (5.3 g) are obtained. [α]^{twenty four} _D+143.3゜(c=1, CH₃OH) Elemental analysis: C₁₃H₂₀NO._Five Calculated value (%): C, 60.92; H, 7.87 Experimental value (%): C, 60.82; H, 8.09 NMR (CDCl)₃) δ: 1.31 (3H, d, J = 7Hz),
1.2~2.0 (10H, m), 3.57 (3H, s), 3.78
(1H, dd, J = 3Hz, 11Hz), 4.16 (1H,
q, J = 7Hz), 4.79 (1H, d, J = 11
Hz), 5.19 (1H, d, J = 3Hz) IR νnujiyor maxcm^-11760 (C=0) Reference example 11 Methyl 6-O-acetyl-2,3-O-cyc
lohexylidene-α-D-glucopyranoside Methyl 4-O-benzoyl-6-bromo-6
-deoxy-α-D-glucopyranoside (27g)
Dissolve in dimethylformamide (50ml) and add 1,
1-dimethoxycyclohexane (40ml) and p-t
Add luenesulfonic acid (1.4g) and aspirate
At 55°C under a vacuum of 40-45 mmHg with vacuum suction.
Stir for 3 hours. Add the reaction solution to ethyl acetate (600ml)
and water (300ml), and add the ethyl acetate layer.
Separate, wash with saturated sodium bicarbonate solution,
After drying with sodium sulfate, concentrate under reduced pressure. residue
Tetrahydrofuran (350ml), methanol
(200ml) and 1N sodium hydroxide (100ml)
Dissolve in the mixed solution and stir at room temperature for 1 hour, then reduce the reaction solution.
Concentrate under pressure. Add water (250ml) to the concentrate and
The resulting oil is extracted with ethyl acetate. Ethyl acetate
Wash the extract with water, dry with sodium sulfate, and reduce pressure.
When concentrated, methyl 2,3-O-cyclohexyly
Den-6-bromo-6-deoxy-α-D-glue
Copyranoside syrup (20g) was obtained.
Ru. The resulting syrupy substance (20g) was dissolved in dimethyl
Sodium acetate dissolved in formamide (250ml)
(14 g) and stirred at 80-85°C for 40 hours.
The reaction solution was concentrated under reduced pressure, and the residue was diluted with ethyl acetate (600
ml) and water (300 ml), plus ethyl acetate.
Separate the layer and add 1N hydrochloric acid and saturated sodium bicarbonate.
After washing with solution and drying with sodium sulfate, concentrate under reduced pressure.
Shrink. Transfer the residue to a column of silica gel (500ml)
Chromatograph the column with toluene-ethyl acetate.
(5:1). After concentrating the eluted fraction under reduced pressure,
When dried under vacuum overnight in a desiccator, the methyl
6-O-acetyl-2,3-O-cyclohexy
Syrup of Lyden-α-D-glucopyranoside
Substance (10.5 g) is obtained. [α]^{twenty four} _D+97.2゜(c=1, CH₃OH) Elemental analysis: C₁₅H_{twenty four}O₇ Calculated value (%): C, 56.95; H, 7.65 Experimental value (%): C, 57.34; H, 7.87 NMR (CDCl)₃) δ: 1.2 to 1.9 (10H, m), 2.11
(3H, s), 3.14 (1H, d, J=3.3Hz),
3.46 (3H, s), 3.4~4.65 (6H, m), 5.06
(1H, d, J=3Hz) Reference example 12 Methyl 6-O-acetyl-2,3-O-cyc
Lohexylidene-α-D-xylo-hexopyrano
Sid-4-ulose (methyl 6-O-acetyl
-2,3-O-cyclohexylidene-α-D-k
scyllo-4-hexopyranose uroside) Dichloromethane in trifluoroacetic anhydride (13.9 ml)
(50 ml) solution in dimethyl sulfoxide (9.3
ml) in dichloromethane (50 ml) at -65℃ or below.
while cooling, and stirred at the same temperature for 10 minutes. child
Add methyl 6-O to the reaction solution while cooling to -70℃ or below.
-acetyl-2,3-O-cyclohexylidene-
α-D-glucopyranoside (10.4g) in dichloro
Add methane (70ml) solution dropwise and stir at the same temperature for 1 hour.
After stirring, add triethylamine while cooling to below -65℃.
(27.6ml) in dichloromethane (40ml) dropwise.
do. Remove the cooling bath until the reaction temperature reaches 20℃.
After stirring, add the reaction solution to ice water. dichloromethane
The layers were separated, the aqueous layer was extracted with dichloromethane, and the aqueous layer was extracted with dichloromethane.
Collect the chloromethane extracts, add 2N hydrochloric acid and sat.
Wash with sodium bicarbonate solution and sodium sulfate
After drying with a vacuum cleaner, concentrate under reduced pressure. Silica gel residue
(400ml) column chromatography, toluene-vinegar
Elute with ethyl acid (5:1). Evacuate the elution fraction
Concentrate and leave the residue under vacuum in a desiccator overnight.
When dried, methyl 6-acetyl-2,3-O-
Cyclohexylidene-α-D-xylo-hexopi
Ranosid-4-ulose (6g) is syrupy
obtained as a quality. [α]^{twenty four} _D+120.4゜(c=1, CH₃OH) Elemental analysis: C₁₅H_{twenty two}O₇ Calculated value (%): C, 57.31; H, 7.06 Experimental value (%): C, 57.60; H, 7.32 NMR (CDCl)₃) δ: 1.2 to 1.85 (10H, m), 2.04
(3H, s), 3.56 (3H, s), 3.82 (1H, dd,
J=3Hz, 11Hz), 4.15~4.7 (3H, m),
4.78 (1H, d, J = 11Hz), 5.27 (1H, d,
J=3Hz) IR νnujiyor maxcm^-1:1765, 1750 Reference example 13 4,7-O-benzylidene-N-benzyloxy
Cycarbonylvalidamine N-benzyloxycarbonylvalidamine (Special
(20g)
was suspended in benzaldehyde (50 ml) and added with sodium chloride.
Add lead (10 g) and stir at room temperature for 5 hours. reaction
Add the solution to a mixture of ethyl acetate and water and add ethyl acetate.
Separate the layers and wash with saturated sodium bicarbonate solution.
After drying with sodium sulfate, concentrate under reduced pressure. Residue
Add petroleum ether (800ml) to the distillate and let it stand at room temperature overnight.
put. Collect the resulting oil and apply it to silica gel.
(550ml) column chromatography, toluene-a
Elute with setone (2:1). Concentrate the eluted fraction under reduced pressure.
When petroleum ether is added to the residue, 4,7-
O-Benzylidene-N-benzyloxycarbonylate
Rubalidamine (21.1 g) is obtained. [α]^{twenty four} _D+54.1゜(c=1, CH₃OH) Elemental analysis: C_{twenty two}H_{twenty five}NO.₆ Calculated value (%): C, 66.15; H, 6.31; N, 3.51 Experimental value (%): C, 66.09; H, 6.16; N, 3.28 NMR (DMSO−d₆) δ: 0.8 to 2.2 (3H, m),
3.1~4.2 (6H, m), 4.70 (1H, d, J=5
Hz), 4.78 (1H, d, J = 4.5Hz), 5.02
(2H, s), 5.47 (1H, s), 6.98 (1H, d,
J=7.5Hz), 7.2~7.6 (10H, m) Reference example 14 2,3-di-O-acetyl-4,7-O-ben
Zylidene-N-benzyloxycarbonylvalida
min 4,7-O-benzylidene-N-benzyloxy
Cycarbonylvalidamine (15g) in pyridine
(150ml) and acetic anhydride (75ml),
Stir overnight at room temperature. Concentrate the reaction solution under reduced pressure to remove the remaining
Add the substance to a mixture of ethyl acetate and water to make ethyl acetate.
Separate the layers. The ethyl acetate layer was treated with 2N hydrochloric acid and saturated charcoal.
Wash with sodium oxyhydrogen solution and sodium sulfate
After drying, concentrate under reduced pressure and add petroleum ether to the residue.
When added, 2,3-di-O-acetyl-4,7-O
-benzylidene-N-benzyloxycarbonyl
Validamine (18.5 g) is obtained. [α]^{twenty four} _D+18.5゜(c=1, CH₃OH) Elemental analysis: C₂₆H₂₉NO.₈ Calculated value (%): C, 64.58; H, 6.05; N, 2.90 Experimental value (%): C, 64.56; H, 6.13; N, 2.81 NMR (CDCl)₃) δ: 1.94 (3H, s), 2.02 (3H,
s), 3.44 (1H, dd, J=3Hz, 10Hz),
3.58 (1H, dd, J=4.5Hz, 10Hz), 4.12
(1H, dd, J=4.5Hz, 11Hz), 4.35 (1H),
4.95 (1H, dd, J=4Hz, 10Hz), 5.10
(2H, s), 5.37 (1H, t, J=10Hz),
5.47 (1H, s), 7.37 (10H, s) Reference example 15 2,3-di-O-acetyl-N-benzyloxy
Cycarbonylvalidamine 2,3-di-O-acetyl-4,7-O-ben
Zylidene-N-benzyloxycarbonylvalida
Dissolve min (18g) in methanol (440ml),
Add 0.5N hydrochloric acid (130ml) and heat under reflux for 30 minutes.
Ru. After cooling the reaction solution to room temperature, add saturated sodium bicarbonate to
Adjust the pH to 5.0 with a diuretic solution and concentrate under reduced pressure. residual
After adding the substance to a mixture of ethyl acetate and water and distributing it, add vinegar
Separate the ethyl acid layer, wash with water, and dilute with sodium sulfate.
After drying, concentrate under reduced pressure. Ethyl ether to the residue
-Adding petroleum ether (1:5) (approximately 1)
2,3-di-O-acetyl-N-benzyloxy
Carbonylvalidamine (11.0g) is obtained [α]^{twenty four} _D+54.8゜(c=1, CH₃OH) Elemental analysis: C₁₉H_{twenty five}NO.₈ Calculated value (%): C, 57.71; H, 6.37; N, 3.54 Experimental value (%): C, 57.67; H, 6.49; N, 3.67 NMR (CDCl)₃) δ: 5.09 (2H, s), 7.36 (5H,
s) Reference example 16 2,3-di-O-acetyl-N-benzyloxy
cycarbonyl-7-O-tritylvalidamine 2,3-di-O-acetyl-N-benzyloxy
Cycarbonylvalidamine (11.0g) in pyridine
(180ml) and add trityl chloride (15g).
and stir overnight at room temperature. The reaction solution was mixed with ethyl acetate and water.
In addition, separate the ethyl acetate layer and add 5%
acid and 5% sodium bicarbonate solution, and sulfuric acid.
After drying with sodium chloride, concentrate under reduced pressure. residue
Subjected to silica gel (450ml) column chromatography,
Elute with toluene-ethyl acetate (3:1). melt
The extracted fraction was concentrated under reduced pressure, and the residue was diluted with ethyl ether.
Add petroleum ether (1:2) and store in the refrigerator overnight.
When left standing, 2,3-di-O-acetyl-N-ben
Zyloxycarbonyl-7-O-tritylvalida
Crystals of min (11.4 g) are obtained. [α]^{twenty five} _D+57.6゜(c=1,DMF) Elemental analysis: C₃₈H₃₉NO.₈ Calculated value (%): C, 71.57; H, 6.16; N, 2.20 Experimental value (%): C, 71.74; H, 6.22; N, 2.12 NMR (CDCl)₃) δ: 1.87 (3H, s), 2.03 (3H,
s), 2.98 (1H, d, J=5Hz), 3.0~3.8
(3H, m), 4.15-4.45 (1H, m), 4.82
(1H, dd, J = 4Hz, 10Hz), 5.07 (2H,
s), 5.17 (1H, t, J = 10Hz), 7.15~
7.55 (20H, m) Reference example 17 (2R)-(2,6/3,4)-2,3-diacetate
xy-4-benzyloxycarbonylamino-6
-trityloxymethylcyclohexanone 2,3-di-O-acetyl-N-benzyloxy
cycarbonyl-7-O-tritylvalidamine
(6.4g) with dimethyl sulfoxide (40ml) and anhydrous
Dissolve in a mixture of acetic acid (20 ml) and stir at room temperature for 15 hours.
Stir. Add the reaction solution to a mixture of ice water and ethyl acetate.
, separate the ethyl acetate layer, and dilute the aqueous layer with ethyl acetate.
Extract. Collect the ethyl acetate extract and add 5% phosphoric acid.
and 5% sodium bicarbonate solution, and sulfuric acid.
After drying with sodium, the solvent is distilled off under reduced pressure. residual
Ethyl ether-petroleum ether (1:20)
(315ml) is added (2R)-(2,6/3,4)-
2,3-diacetoxy-4-benzyloxycar
Bonylamino-6-trityloxymethylcyclo
Hexanone (5.7 g) is obtained. [α]^{twenty five} _D+56.4゜(c=1,DMF) Elemental analysis: C₃₈H₃₇NO.₈ Calculated value (%): C, 71.39; H, 5.87; N, 2.20 Experimental value (%): C, 71.71; H, 5.66; N, 2.08 NMR (CDCl)₃) δ: 1.4-2.8 (3H, m), 1.94
(3H, s), 2.08 (3H, s), 3.1~3.6 (2H,
m), 4.4-4.7 (1H, m), 5.05 (1H, dd,
J=4Hz, 12Hz), 5.12 (2H, s), 5.43
(1H, d, J = 12Hz), 5.77 (1H, d, J
=8Hz), 7.15~7.55 (20H, m) Reference example 18 N,N'-diethoxycarbonyl-4,5-O
-isopropylidene-6-oxo-2-deoxy
Streptamine and N,N'-diethoxylic
Bonyl-5,6-O-isopropylidene-4-o
Mixture of xo-2-deoxystreptamine N,N'-diethoxycarbonyl-4,5-O
-isopropylidene-2-deoxystreptami
and N,N'-diethoxycarbonyl-5,6-
O-isopropylidene-2-deoxystrepta
Mixture of mint (11.0g) in chloroform (100ml)
Dissolve in water, add water (90 ml), and add water while stirring.
Potassium urate (15.0g) and potassium carbonate (15.0g)
g), then add ruthenium dioxide (400mg) and leave in the chamber.
Stir at room temperature for 15 hours. 2-propanol in the reaction solution
(10 ml) and stir at room temperature for 30 minutes. reaction mixture
The compound is filtered off and washed with chloroform. Filtrate
Combine the washing solution and the washing solution, separate the chloroform layer, and add water.
Extract the layers with dichloromethane. Combine the organic solvent layers.
After drying with magnesium sulfate, remove the solvent under reduced pressure.
and add ethyl ether (30 ml) and petroleum ether to the residue.
Add gelatin (100ml) and leave it in the refrigerator overnight.
Then, crystals (8.0 g) of the oxo derivative are obtained. Melting point: 184℃ (decomposition) Elemental analysis: C₁₅H_{twenty four}N₂O₇ Calculated value (%): C, 52.32; H, 7.03; N, 8.14 Experimental value (%): C, 52.08; H, 7.13; N, 7.82 NMR (CDCl)₃) δ: 1.23 (6H, t, J=7Hz),
1.47 (6H, s), 2.75-3.15 (1H, m), 3.64
(1H, t, J = 10Hz), 4.01 (2H, q, J
= 7Hz), 4.03 (2H, q, J = 7Hz), 4.37
(1H, dd, J = 1Hz, 10Hz), 5.05 (1H,
d, J = 7.5Hz), 5.56 (1H, d, J = 7.5
Hz) Reference example 19 (1R)−(1(CH₂OH), 2, 4, 5/1 (OH),
3)-5-amino-1-hydroxymethyl-1,
2,3,4-cyclohexanetetrol (vario
(5-epiisomer of amine) (1R)-(1,3,4/2)-tri-O-acetyl
-4-benzyloxycarbonylamino-6-
Exomethylene-1,2,3-cyclohexane
Riolu (Patent application 144309, 1984, page 35, reference example 5)
Dissolve (3.8g) in ethyl ether (50ml)
Add pyridine (3 ml) and cool in an ice water bath.
of osmium tetroxide (2.4 g) in ethyl ether
Add solution (20ml) dropwise. The reaction solution was kept at room temperature for 4 hours.
After stirring, leave at room temperature overnight. The reaction solution was diluted with ice water.
After cooling for an hour, the resulting precipitate was collected by filtration and cooled with ethyl ether.
After washing with towel, dry. Obtained precipitate (2.8g)
Dissolve in ethanol (50 ml), water (100 ml) and then
Add sodium sulfite (14g) at room temperature for 20 minutes.
Stir for an hour. The resulting black precipitate was filtered off and evaporated.
Wash with alcohol, combine the filtrate and washings, and reduce the solvent.
Distill under pressure. The residue was dissolved in a mixture of ethyl acetate and water.
Add and partition, separate the ethyl acetate layer, and remove the aqueous layer.
Extract three times with ethyl acetate. Combine the ethyl acetate layers.
After washing with water and drying with sodium sulfate, concentrate under reduced pressure.
Then, (1R)−(1(CH₂OH), 2, 4, 5/1
(OH),3)-2,3,4-tri-O-acetyl
-5-benzyloxycarbonylamino-1-hyron
Droxymethyl-1,2,3,4-cyclohexa
Ntetrol was obtained as an oil (2.0 g).
Ru. This oily substance was mixed with methanol-water-acetic acid (1:
1:0.2) (70ml) and palladium black (500ml).
mg) and stirred at room temperature in a hydrogen stream for 2.5 hours.
Remove the catalyst by filtration, wash with 50% methanol water, and collect the filtrate.
Combine the washing liquid and concentrate to dryness under reduced pressure. residue
Dissolve in 1N sodium hydroxide solution (100ml) and bring to room temperature.
Stir at warm temperature for 3 hours. The reaction solution was adjusted to pH5 with 2N hydrochloric acid.
Adjust, Amberlight CG-50 (NH⁺ _Fourtype, low
Mu & Haas, USA) (250ml)
After washing the column with water, add 0.2N ammonia.
Elute with water, concentrate the eluted fraction under reduced pressure, and freeze-dry.
When dried, (1R)-(1(CH₂OH), 2, 4, 5/1
(OH),3)-5-amino-1-hydroxymethylene
Ru-1,2,3,4-cyclohexanetetrol
of white powder (400 mg) is obtained. [α]^{twenty three} _D+17.5゜(c=1,H₂O) Elemental analysis: C₇H₁₅NO._Five・H₂O Calculated value (%): C, 39.80; H, 8.11; N, 6.63 Experimental value (%): C, 40.11; H, 8.04; N, 6.92 NMR(D₂O) δ: 1.88 (1H, dd, J=4.5Hz,
14.5Hz), 2.14 (1H, dd, J=7Hz, 14.5
Hz), 3.5-4.1 (5H, m), 4.17 (1H, t,
J=6.5Hz) TLC: Rf=0.42 Reference example 20 3-O-benzyloxycarbonyl-4,7-
O-isopropylidenevalienamine 1,2-ka
Rubamate a N-benzyloxycarbonylvalienamine
(20g) in 1N sodium hydroxide solution (200ml)
and stir at room temperature for 2 hours. reaction solution
Adjust the pH to 5.5 with 2N hydrochloric acid and wash with ethyl ether.
After cleaning, concentrate under reduced pressure. Activate the residue (400ml)
After washing with water, 50% methanol
Elute with cool water. After concentrating the eluate under reduced pressure, freeze it.
When dried, valienamine 1,2-carbame
(12 g) is obtained. b Valienamine 1,2-carbamate (10
Dissolve g) in dimethylformamide (150ml)
and 2,2-dimethoxypropane (20 ml) and p
-Add toluenesulfonic acid (250mg) and heat at 100℃
Stir for 2 hours. After cooling the reaction solution to room temperature,
X1×2 (OH^-Mold, Dow Chemical Company
Add about 50 ml of the product (manufactured in the United States) and stir for 30 minutes.
Filter off Dowex 1x2 and wash with ethanol.
Purify. The filtrate and washing liquid were combined and concentrated under reduced pressure.
Add toluene to the residue and repeat concentration under reduced pressure.
vinegar. Dissolve the residue in water (approximately 50 ml) and add to the MCI gel.
Le CHP-20P (manufactured by Mitsubishi Chemical Industries, 400ml) color
After washing the column with water, water-methano
Elute with a gradient of
After concentration and lyophilization, 4,7-O-isopropylene
Pylidenevalienamine 1,2-carbamate
(11.3g) is obtained. c 4,7-O-isopropylidenevalienamine
1,2-carbamate (4.8g) in dimethyl
Dissolve in formamide (50ml) and cool below -40℃
Cool, benzyloxycarbonyl chloride
(5.1ml), then triethylamine (3.1ml)
Add and stir at -20 to -10°C for 2 hours. reaction solution
of 2N hydrochloric acid (500ml) and ethyl acetate (500ml)
Add to the mixture under ice cooling and separate the ethyl acetate layer.
After washing with water and drying with sodium sulfate, concentrate under reduced pressure.
do. Transfer the residue to a column of silica gel (250ml)
Chromatographed toluene-ethyl acetate (1:
Elute with 1). Concentrate the eluted fraction under reduced pressure to remove the remaining
Ethyl ether-petroleum ether (1:10)
(approximately 300ml) and leave it in the refrigerator overnight.
3-O-benzyloxycarbonyl-4,7-
O-isopropylidenevalienamine 1,2-
Carbamate crystals (4.3 g) are obtained. [α]^{twenty four} _D+126.8゜(c=1, CH₃OH) Elemental analysis: C₁₉H_{twenty one}NO.₇ Calculated value (%): C, 60.79; H, 5.64; N, 3.37. Experimental value (%): C, 60.68; H, 5.48; N, 3.51. Reference example 21 (1R)−(1−(CH₂OH), 2, 4, 5/1
(OH),3)-5-amino-1-hydroxymethylene
Ru-1,2,3,4-cyclohexanetetrol
(5-epiisomer of variolamine) 3-O-benzyloxycarbonyl-4,7-
O-isopropylidenevalienamine 1,2-ka
Rubamate (3.7g) in acetonitrile (60ml)
Add bromine to the solution, while stirring, while cooling with ice water.
(1.8g) in acetonitrile (20ml) dropwise.
and stir for an additional 5.5 hours. Filter the resulting precipitate,
Wash with a small amount of acetonitrile and ethyl ether.
Purify. The obtained powder (1.2g) was mixed with 50% ethanol.
Hydrogenated fluoride was dissolved in water (20 ml) and cooled with ice water.
Add sodium urethane (500 mg) and leave at room temperature for 4 hours.
Stir. Concentrate the reaction solution under reduced pressure and add activated carbon (180ml).
After column chromatography and washing the column with water, 50%
Elute with methanol water. Concentrate the eluate under reduced pressure,
Add barium hydroxide to the resulting aqueous solution (approximately 20 ml)
(2.0g) and heated under reflux for 2 hours. reaction solution
After cooling to room temperature, carbon dioxide gas is passed through the resulting precipitate.
Filter off the lees. Filter the filtrate with Amberlite CG-50
(NH⁺ _FourModel, manufactured by Rohm and Haas, USA)
(250ml) column chromatography and wash the column with water.
Then, elute with 0.2N ammonia water. elution fraction
After concentration under reduced pressure, lyophilization produces variolamine.
A 5-epiisomer white powder (120 ml) is obtained. Reference example 22 6,7,8-triacetoxy-1-azidomethy
Ru-3-oxo-2-oxa-4-azabicyclo
[3.3.1] Nonane a 6,7,8-triacetoxy-1-bromome
Chil-3-oxo-2-oxa-4-azabisi
Black [3.3.1] Nonane (patent application 144309, 37
page, described in Example 1) (4.1 g) was dissolved in dimethylform.
Sodium azide dissolved in Muamide (15ml)
(1.0g) and stirred for 7 hours at a bath temperature of 130-140℃.
do. The reaction solution was concentrated under reduced pressure and further mixed with toluene.
Dimethylformamide is distilled off under reduced pressure while boiling.
Add 20% methanol water (100ml) to the residue,
Leave in the refrigerator overnight. Filter the formed crystals
After washing with ice-cold 20% methanol water, dry.
Then, 6,7,8-triacetoxy-1-azo
Domethyl-3-oxo-2-oxa-4-aza
Bicyclo[3.3.1]nonane (2.7 g) is obtained. [α]^{twenty three} _D+79.1゜(c=1,DMF) IR ν^KBr _naxcm^-1:2130 Elemental analysis: C₁₄H₁₈N_FourO₈ Calculated value (%): C, 45.40; H, 4.90; N,
15.13. Experimental value (%): C, 45.58; H, 4.88; N,
15.31. b 6,7,8-triacetoxy-1-iodomethane
Chil-3-oxo-2-oxa-4-azabisi
Black [3.3.1] Nonane (Patent application 144309, 1987, 39)
page, described in Example 2) (4.5 g) in dimethylform.
Sodium azide dissolved in Muamide (15ml)
(1.0g) and stirred for 3 hours at a bath temperature of 130-140℃.
do. Treat the reaction solution in the same manner as in Reference Example a) above.
and 6,7,8-triacetoxy-1-azide
Methyl-3-oxo-2-oxa-4-azabi
Cyclo[3.3.1]nonane crystals (3.2g) were obtained.
It will be done. Reference example 23 7-azido-7-deoxyvariolamine 6,7,8-triacetoxy-1-azidomethy
Ru-3-oxo-2-oxa-4-azabicyclo
[3.3.1] Suspend nonane (1.0 g) in water (100 ml),
Add barium hydroxide (4.0g) and heat at 70℃ for 5 hours
Stir. After cooling the reaction solution to room temperature, carbon dioxide gas was added.
The precipitate formed is filtered off and washed with water. filtrate
Combine the washing liquid and Amberlite CG-50 (NH⁺ _Four
Mold, Rohm and Haas, USA) (250
ml) column chromatography, and after washing the column with water,
Elute with 0.1N ammonia water. Evacuate the elution fraction
Concentrate and transfer the residue to Dowex 1×2 (OH^-type,
Dow Chemical Co., USA) (250ml) Column
chromatograph and elute with water. Concentrate the eluted fraction under reduced pressure.
After shrinkage and lyophilization, 7-azido-7-deoxy
Variolamine (370ml) is obtained. [α]^{twenty three} _D+43.1゜(c=1,H₂O) IR ν^KBr _naxcm^-1:2115 Elemental analysis: C₇H₁₄N_FourO_Four Calculated value (%): C, 38.53; H, 6.47; N,
25.68. Experimental value (%): C, 38.71; H, 6.37; N,
25.45. NMR(D₂O) δ: 1.78 (1H, dd, J=3.8Hz,
15Hz), 2.15 (1H, dd, J=3Hz, 15Hz),
3.44 (1H, d, J = 13.5Hz), 3.4~3.6 (1H,
m), 3.52 (1Hd, J=9.5Hz), 3.65 (1H,
d, J = 13.5Hz), 3.73 (1H, dd, J = 4.5
Hz, 9.5Hz), 3.99 (1H, t, J = 9.5Hz). TLC: Rf=0.54 Reference example 24 7-amino-7-deoxyvariolamine 7-azido-7-deoxyvariolamine
(250ml) in 2% acetic acid water (50ml),
Add Umu Kuro (200 mg) and incubate for 3 hours in a hydrogen stream at room temperature.
Stir for a while. The catalyst was removed by filtration, and the reaction solution was concentrated to dryness under reduced pressure.
After solidification, the aqueous solution of the residue was mixed with Amberlite CG-50.
(NH⁺ _Fourmold, manufactured by Rohm, & Haas, USA)
(180ml) column chromatography. Column with water
After washing with 0.2N ammonia water, wash with 0.5N ammonia water.
Elutes with After concentrating the eluate fraction under reduced pressure, freeze-dry it.
and 7-amino-7-deoxyvariolamine
(180g) is obtained. [α]^{twenty three} _D+20.1゜(c=1,H₂O) Elemental analysis: C₇H₁₆N₂O_Four・H₂O Calculated value (%): C, 39.99; H, 8.63; N,
13.33. Experimental value (%): C, 39.85; H, 8.25; N,
12.76. NMR(D₂O) δ: 1.73 (1H, dd, J=3.5Hz,
15Hz), 2.07 (1H, dd, J=3Hz, 15Hz),
2.77 (1H, d, J = 13.5Hz), 2.96 (1H,
d, J=13.5Hz), 3.4~3.85 (3H, m),
3.99 (1H, t, J = 9.5Hz). TLC: Rf=0.14 Reference example 25 6,7,8-triacetoxy-1-methyl-3
-oxo-2-oxa-4-azabicyclo
[3.3.1] Nonane 6,7,8-triacetoxy-1-bromomethy
Ru-3-oxo-2-oxa-4-azabicyclo
[3.3.1] Nonane (Patent Application 1987-144309, p. 37, carried out
(described in Example 1) (800mg) dissolved in toluene (50ml)
and tri-n-butyltin hydride (1 ml)
and α,α′-azo-bis-isobutyronitrile
(100 mg) and heated to reflux while stirring for 1 hour.
The reaction solution was concentrated under reduced pressure, and petroleum ether was added to the residue.
Leave it in the refrigerator overnight. Filter the formed precipitate
After drying, silica gel column chromatography (180
ml) and elute with ethyl acetate. elution fraction
Concentrate under reduced pressure and add ethyl ether-petroleum ether to the residue.
Add Teru (1:1) and leave it in the refrigerator overnight.
and 6,7,8-triacetoxy-1-methyl-3
-oxo-2-oxa-4-azabicyclo
[3.3.1] Nonane crystals (510 mg) are obtained. [α]^{twenty four} _D+61.3゜(c=1,H₂O) Elemental analysis: C₁₄H₁₉NO.₈ Calculated value (%): C, 51.06; H, 5.82; N, 4.25. Experimental value (%): C, 51.03; H, 5.76; N, 4.19. NMR (DMSO−d₆) δ: 1.17 (3H, s), 1.93
(3H, s), 1.98 (3H, s), 2.03 (3H,
s), 3.4-3.6 (1H, m), 4.8-5.2 (3H,
m), 7.77 (1H, d, J = 5.5Hz). Reference example 26 7-deoxyvariolamine 6,7,8-triacetoxy-1-methyl-3
-oxo-2-oxa-4-azabicyclo
[3.3.1] Suspend nonane (350 mg) in water (50 ml),
Add barium hydroxide (2.0g) and heat at 90-100℃
Stir for an hour. After cooling the reaction solution to room temperature, carbon dioxide
The resulting precipitate is filtered off and washed with water.
Combine the filtrate and washing liquid and use Amberlite CG-50.
(NH⁺ _Fourmold, manufactured by Rohm, & Haas, USA)
(150ml) column chromatography and wash the column with water.
After that, elute with 0.2N ammonia water and reduce the eluted fraction.
Concentrate under pressure. Dowex 1×2 (OH^-
Mold, Dow Chemical Co., USA) (250ml)
Apply to muchromatography and elute with water. Reduce elution fraction
After pressure concentration and lyophilization, 7-deoxyvariol
Ruamine (140 mg) is obtained. [α]^{twenty three} _D+12.3゜(c=1,H₂O) Elemental analysis: C₇H₁₅NO._Four・H₂O Calculated value (%): C, 43.07; H, 8.78; N, 7.18. Experimental value (%): C, 43.31; H, 8.59; N, 7.11. NMR(D₂O) δ: 1.43 (3H, s), 1.90 (1H,
dd, J=3.8Hz, 15.5Hz), 2.17(1H, dd,
J=2.6Hz, 15.5Hz), 3.45(1H, d, J=
9Hz), 3.45-3.7 (1H, m), 3.85 (1H,
dd, 4Hz, 9.2Hz), 4.00 (1H, t, J=9
Hz). I C₅₀(Satsukarase): 2.4×10^-FiveM TLC: Rf=0.34 Reference example 27 6,7,8-trihydroxy-1-hydroxy
Methyl-3-oxo-2-oxa-4-azatri
Cyclo [3.3.1.0^4,9] Nonane 9-bromo-6,7,8- obtained in Reference Example 3
trihydroxy-1-hydroxymethyl-3-o
Xo-2-oxa-4-azabicyclo [3.3.1]
Dissolve nonane (5.0g) in methanol (100ml)
Add sodium hydrogen carbonate (2.5g) and leave for 2 hours.
Heat to reflux while stirring for a while. Insoluble matter is filtered off, and meth
Wash with alcohol, collect the filtrate and washings, and concentrate under reduced pressure.
Ru. Transfer the residue to a column of MCI gel CHP20P (500ml)
Chromatograph and elute with water. Elution fraction (400
After concentrating ~600ml) under reduced pressure and freeze-drying, 6,
7,8-trihydroxy-1-hydroxymethyl
-3-oxo-2-oxa-4-azatricyclo
[3.3.1.0^4,9] Nonane (2.8 g) is obtained. [α]²⁶ _D+74.7゜(c=1,H₂O) IR ν^KBr _naxcm^-1:1780, 1740. Elemental analysis: C₁₈H₁₁NO.₆ Calculated value (%): C, 44.24; H, 5.11; N, 6.45. Experimental value (%): C, 44.20; H, 5.02; N, 6.43. Reference example 28 1-hydroxymethyl-5,6-epiminosic
Lohexane-1,2,3,4-tetrol (1,
6-epiminovariolamine) 6,7,8-trihydroxy-1-hydroxy
Methyl-3-oxo-2-oxa-4-azabisi
Kuro [3.3.1.0^4,9] Nonane (2.0g) water (200ml)
Add barium hydroxide (8.0g) to 60
Stir at ~70°C for 5 hours. Cool the reaction solution to room temperature
After that, barium carbonate is generated through carbon dioxide gas.
The precipitate is filtered and washed with water. Combine the filtrate and washing liquid.
Amberlight CG-50 (NH⁺ _FourType, Rohm
Column and Haas Co., USA) (250ml)
After washing the column with water, add 0.1N ammonia
Elute with water. Concentrate the eluted fraction under reduced pressure and remove the residue.
Dowex 1×2 (OH^-mold, dow chemical
Co., Ltd., USA) (500ml) column chromatography.
Elutes with water. Concentrate the eluted fraction under reduced pressure to obtain a residue.
If you add methanol and leave it in the refrigerator overnight, 1
-hydroxymethyl-5,6-epiminocyclohe
Xane-1,2,3,4-tetrol (1,6-
Epiminovariolamine) crystals (385mg)
can get. [α]^{twenty three} _D−4.8° (c=1, H₂O) Elemental analysis: C₇H₁₃NO._Five Calculated value (%): C, 43.97; H, 6.85; N, 7.33. Experimental value (%): C, 43.71; H, 6.90; N, 7.23. TLC: Rf=0.44 Reference example 29 N-benzyloxycarbonyl-7-O-tri
tilvalidamine N-benzyloxycarbonylvalidamine (Special
Application No. 56-144309, page 30, described in Reference Example 1) (3.1
Dissolve g) in pyridine (20ml) and add trityl chloride.
(3.5 g) and stirred at room temperature overnight. reaction solution
Concentrate under reduced pressure and extract the residue with ethyl acetate.
The solution was diluted with 5% phosphoric acid and 5% sodium bicarbonate solution.
After washing and drying with sodium sulfate, concentrate under reduced pressure.
Ru. Column chromatography of the residue on silica gel (400 ml)
chloroform-methanol (9:
Elute with 1). Concentrate the eluted fraction under reduced pressure and remove the residue.
When ethyl ether is added to N-benzyloxy
Crystals of carbonyl-7-O-tritylvalidamine
(3.3g) is obtained. IR ν^KBr _naxcm^-1:3400,1700 Reference example 30 N-benzyloxycarbonyl-2,3,4-
tri-O-acetyl-7-O-tritylvalidami
hmm N-benzyloxycarbonyl-7-O-tri
Tilvalidamine (3.0g) to pyridine (30ml)
Dissolve, add acetic anhydride (15 ml) and stir overnight at room temperature.
After concentrating the reaction solution under reduced pressure, the residue was diluted with ethyl acetate.
Dissolved in 5% phosphoric acid and 5% sodium bicarbonate.
After washing with silica solution and drying with magnesium sulfate,
Concentrate under pressure. Pour the residue into silica gel (400ml).
Lamb chromatography and toluene-ethyl acetate
(5:1). Concentrate the eluted fraction under reduced pressure,
Ethyl ether-petroleum ether (1:
10), N-benzyloxycarbonyl-
2,3,4-tri-O-acetyl-7-O-tri
Tiluvalidamine (3.5 g) is obtained. Elemental analysis: C₄₀H₄₁NO.₉ Calculated value (%): C, 70.67; H, 6.08; N, 2.06 Experimental value (%): C, 70.45; H, 6.05; N, 2.34 IR ν^KBr _naxcm^-1:3340, 1750, 1730 Reference example 31 N-benzyloxycarbonyl-2,3,4-
Tri-O-acetylvalidamine N-benzyloxycarbonyl-2,3,4-
tri-O-acetyl-7-O-tritylvalidami
(3.0 g) in 80% acetic acid (60 ml) (60-80 mL).
℃) to dissolve and stir at 80℃ for 1 hour. reaction solution
was concentrated to dryness under reduced pressure, and the residue was extracted with ethyl acetate.
Then, add the ethyl acetate extract to 5% phosphoric acid and 5% carbonated water.
Sodium solution, and washed with water, sodium sulfate
After drying with aluminum, it was concentrated to dryness under reduced pressure, and the residue was
The precipitate formed by adding ether is filtered and dried.
The obtained powder (2.3 g) was dissolved in chloroform,
Subjected to silica gel (300ml) column chromatography,
Elute with chloroform-methanol (9:1)
Ru. The eluted fraction was concentrated to dryness under reduced pressure to yield N-benzyl.
Oxycarbonyl 2,3,4-tri-O-acetyl
Rubalidamine (1.8 g) is obtained. IR ν^KBr _naxcm^-1:3440, 3360, 1735 Reference example 32 N-benzyloxycarbonyl-2,3,4-
Tri-O-acetyl-7-deoxy-7-phthal
imidovalidamine N-benzyloxycarbonyl-2,3,4-
Tri-O-acetylvalidamine (2.2g), phthalate
imide (1.1g), and triphenylphosphine
(1.6g) dissolved in tetrahydrofuran (50ml)
and diethyl azodicarboxylate (1.1 g) under ice cooling.
and stirred at the same temperature for 2 hours. Reduce the pressure of the reaction solution
Concentrate and extract the residue with ethyl acetate. extract liquid
was washed with 5% sodium bicarbonate solution and 5% phosphoric acid.
After washing with sodium sulfate, concentrate to dryness under reduced pressure.
Ru. Dissolve the residue in toluene and silica gel
(400 ml) column chromatography, toluene-a
Elute with setone (9:1). Concentrate the eluted fraction under reduced pressure.
When ethyl ether is added to the residue, N-base is obtained.
ndyloxycarbonyl-2,3,4-tri-O
-acetyl-7-deoxy-7-phthalimidova
Lydamine crystals (0.7 g) are obtained. IR ν^KBr _naxcm^-1:3380, 1785, 1735, 1710 Reference example 33 2,3,4-tri-O-acetyl-7-deoxy
Sea-7-phthalimidovalidamine N-benzyloxycarbonyl-2,3,4-
O-triacetoxy-7-deoxy-7-phthal
Imidovalidamine (640mg) in 70% methanol water
(50ml), acetic acid (10ml), and ethyl acetate (10ml).
Add palladium black (215mg) to the mixture and add water.
Stir for 4 hours at room temperature in a stream of bare air. Filter off catalyst
After washing with ethanol, combine the filtrate and washing liquid,
Concentrate under reduced pressure. Add ethyl ether to the residue
and 2,3,4-tri-O-acetyl-7-deoxy
C-7-phthalimidovalidamine (335mg) is obtained.
It will be done. IR ν^KBr _naxcm^-1:1770, 1740, 1715, 1240 Reference example 34 7-Amino-7-deoxyvalidamine 2,3,4-tri-O-acetyl-7-deoxy
C-7-phthalimidovalidamine (250mg) 2
% hydrazine hydrate dissolved in methanol solution (25ml)
and stir overnight at room temperature. Concentrate the reaction solution under reduced pressure,
The residue is Amberlite CG-50 (NH⁺ _Fourtype, row
(manufactured by Mu, & Haas, USA) (200ml)
After applying to muchromatography and washing with water (800ml), 0.4%
Elute with aqueous ammonia. After concentrating the eluted fraction under reduced pressure
When freeze-dried, 7-amino-7-deoxyvari
Damin (64mg) is obtained. [α]^{twenty three} _D+43.3゜(c=1,H₂O) Elemental analysis: C₇H₁₆N₂O₃・2H₂O Calculated value (%): C, 39.61; H, 9.50; N, 13.20 Experimental value (%): C, 39.33; H, 9.75; N, 13.14 TLC: Rf=0.14 Reference example 35 N-ethoxycarbonyl-7-O-tritylba
Lienamine N-ethoxycarbonylvalienamine (5.0g)
was dissolved in pyridine (35 ml) and added to trichloride under ice cooling.
Add chill (11.3 g) and stir at room temperature overnight. anti
The reaction solution was concentrated under reduced pressure, and the residue was dissolved in chloroform (300
ml), and the chloroform solution was dissolved in 2N hydrochloric acid, 5
% sodium bicarbonate solution, and washed with water;
After drying with magnesium sulfate, remove the solvent under reduced pressure.
Ru. Dissolve the residue in ethyl ether and dissolve in petroleum ether.
When ter is added, N-ethoxycarbonyl-7-O
- Tritylvalienamine crystals (4.8g) were obtained.
It will be done. The mother liquor was concentrated to dryness under reduced pressure, and the residue was filtered with chlorofluorophore.
Column solution of silica gel (180 ml)
chloroform-methanol (19:
Elute with the mixture of 1). Concentrate the eluted fraction to dryness.
and the residue was condensed with ethyl ether-petroleum ether.
When crystallized, further N-ethoxycarbonyl-7
-O-tritylvalienamine crystals (3.0g)
can get. IR ν^KBr _naxcm^-1:3400,1705 Reference example 36 N-ethoxycarbonyl-2,3,4-tri-
O-acetyl-7-O-tritylvalienamine N-ethoxycarbonyl-7-O-tritylba
Dissolve lienamine (7.5g) in pyridine (75ml)
Then, add acetic anhydride (37.5 ml) under ice-cooling, and stir at room temperature.
Stir overnight. The reaction solution was concentrated to dryness under reduced pressure, and the residue
was partitioned between ethyl acetate and water and the ethyl acetate layer
5% phosphoric acid, 5% sodium bicarbonate
Wash with solution and water and dry with sodium sulfate
After that, it is concentrated to dryness under reduced pressure. Residue in ethyl ether
When dissolved in and adding petroleum ether, N-ethoxy
carbonyl-2,3,4-tri-O-acetyl-
7-O-tritylvalienamine crystals (9.0g)
is obtained. IR ν^KBr _naxcm^-1:3330, 1750, 1740, 1700 Reference example 37 N-ethoxycarbonyl-2,3,4-tri-
O-acetylvalienamine N-ethoxycarbonyl-2,3,4-tri-
O-acetyl-7-O-tritylvalienamine
(8.5g) in ethyl acetate (80ml) and 80% vinegar.
Add acid water (160 ml) and stir at 60°C for 8 hours.
The reaction solution was concentrated under reduced pressure, and the residue was extracted with ethyl acetate.
Then, add the ethyl acetate extract to 5% phosphoric acid and 5% carbonated water.
Sodium solution, and washed with water, sodium sulfate
After drying with aluminum, concentrate to dryness under reduced pressure. To remove the residue
Column of silica gel (250ml) dissolved in toluene
Chromatographed with toluene-acetone (3:1)
Elutes with The eluted fraction was concentrated to dryness under reduced pressure, and the residue
Dissolve in ethyl ether and add petroleum ether
and N-ethoxycarbonyl-2,3,4-tri
-O-acetylvalienamine crystals (2.8g)
can get. IR ν^KBr _naxcm^-1:3480, 3300, 1740, 1710 Reference example 38 N-ethoxycarbonyl-2,3,4-tri-
O-acetyl-7-deoxy-7-phthalimide
Valienamine N-ethoxycarbonyl-2,3,4-tri-
O-acetylvalienamine (2.5g), phthalimide
(1.95g) and triphenylphosphine
(2.65g) dissolved in tetrahydrofuran (45ml)
and diethyl azodicarboxylate (1.65
ml) at the same temperature for 1 hour and then at room temperature for 30 minutes.
Stir. The reaction solution was concentrated under reduced pressure, and the residue was dissolved in acetic acid ethyl chloride.
Partition between chill and water and separate ethyl acetate layer.
5% phosphoric acid water, 5% sodium bicarbonate solution
After washing with water and drying with sodium acetate, reduce
The solvent is removed under pressure. Dissolve the residue in toluene
Column chromatography (270ml) using silica gel (270ml)
ml) and eluted with toluene-acetone (6:1).
do. The eluted fractions were concentrated under reduced pressure, and the residue was dissolved in ethyl ethyl ethyl ethyl ether.
Add 130ml of nitrogen and leave it in a cool place.
-ethoxycarbonyl-2,3,4-tri-O-
Acetyl-7-deoxy-7-phthalimidovari
Enamine crystals (2.2 g) are obtained. IR ν^KBr _naxcm^-1:3350, 1780 (sh), 1730, 1725 Reference example 39 7-amino-7-deoxyvalienamine N-ethoxycarbonyl-2,3,4-tri-
O-acetyl-7-deoxy-7-phthalimide
Valienamine (2.0g) in 2% hydrazine hydrate
Dissolve in ethanol solution (400ml) and stir overnight at room temperature.
Stir. Add water (400ml) under ice-cooling and adjust the pH with acetic acid.
6, methanol was distilled off under reduced pressure, and insoluble matter was removed.
Filter off. The filtrate was treated with Amberlite CG-50 (NH⁺ _Four
Mold, manufactured by Rohm, & Haas, USA) (800
ml) and washed with water (1.6).
After cleaning, elute with 0.4% ammonia water. Elution fraction
After concentrating under reduced pressure and lyophilizing, 7-amino-7
-deoxyvalienamine (0.3 g) is obtained. [α]^{twenty three} _D+70.1゜(c=1,H₂O) Elemental analysis: C₇H₁₄N₂O₃・2H₂O Calculated value (%): C, 39.99; H, 8.63; N, 13.33 Experimental value (%): C, 39.68; H, 8.79; N, 13.14 TLC: Rf=0.22 Reference example 40 Methyl 4 [(1S,6S)-(4,6/5)-4,
5,6-trihydroxy-3-hydroxymethyl
-2-cyclohexen-1-yl]amino-4,
6-dideoxy-α-D-glucopyranoside and
Biso 6-deoxygalactopyranoside isomer Valienamine (2.0g) and methyl 2,3-O
-cycloexylidene-6-deoxy-α-D-
Xylo-4-hexopyranoside-4-ulose
(Methyl 2,3-O-cyclohexylidene-6
-deoxy-α-D-xylo-4-hexopyrano
-Suuroside) (5.5g) in dimethylformamide
(50 ml), 2N hydrochloric acid (1.5 ml) and
Add sodium borohydride (2.6g),
Stir at ~70 °C for 18 h. Concentrate the reaction solution under reduced pressure,
Dissolve the residue in water (150 ml) and
50W×8(H⁺(Mold, Dow Chemical Company, USA)
(150 ml) and stirred at room temperature for 2 hours. This mixture
Pour the compound into another Dowex 50W x 8 (H⁺type, 30
ml) onto the packed column and wash the column with water.
After (1), elute with 0.5N ammonia water.
The eluted fraction (0.7-1.6) was concentrated under reduced pressure and
Light CG-50 (NH⁺ _FourType, Rome, and Her
Column chromatography (250 ml) manufactured by
and elute with water. Reduce the elution fraction (150-270ml)
Concentrate under pressure and remove the residue by dowering 1×2 (OH^-type,
Dow Chemical Co., USA) (1) Column
When chromatographed and eluted with water, it is separated into two components.
Ru. Reduce pressure on the fractions eluted earlier (1.10-1.71)
After concentration and lyophilization, 6-deoxygalactopi
The ranosidic isomer (370 mg) was obtained and then eluted
The fraction (2.24-4.56) obtained was concentrated under reduced pressure.
Add ethanol (150 ml) to the syrupy substance.
If you leave it in the refrigerator overnight, 6-deoxyglu
Crystals (1.0 g) of the copyranoside isomer are obtained.
When the mother liquor was concentrated to dryness under reduced pressure, an additional 230 mg was recovered.
Ru. The isomer eluted first (6-deoxygalacto
pyranoside isomer): [α]^{twenty two} _D+133.6゜(c=1,H₂O) Elemental analysis: C₁₄H_{twenty five}NO.₈・1/2H₂O Calculated value (%): C, 48.83; H, 7.61; N, 4.07 Experimental value (%): C, 48.58; H, 7.66; N, 3.86 NMR(D₂O) δ: 1.64 (3H, d, J = 6.5Hz),
3.37 (1H, dd, J=1.5Hz, 4Hz), 3.6~
4.7 (9H, m), ~5.1 (1H), 6.32 (1H, J=
1.5Hz, 5Hz) I C₅₀(Satsukarase): 6.6×10^-FiveM TLC: Rf=0.45 The later eluted isomer (6-deoxyglucopi
Ranosidic isomer): [α]^{twenty two} _D+131.5゜(c=1,H₂O) Elemental analysis: C₁₄H_{twenty five}NO.₈・1/2H₂O Calculated value (%): C, 48.83; H, 7.61; N, 4.07 Experimental value (%): C, 48.59; H, 7.56; N, 3.69 NMR(D₂O) δ: 1.68 (3H, d, J = 6.5Hz),
2.88 (1H, m), 3.7-4.75 (9H, m), 3.74
(3H, s), ~5.05 (1H), 6.25 (1H, dd,
J=1.5Hz, 5Hz) I C₅₀(maltase): 3.2×10^-6M I C₅₀(Satsukaraze): 7.0×10^-7M TLC: Rf=0.54 Reference example 41 N-(2,3,5-trihydroxy-6-hydro
roxymethyltetrahydropyran-4-yl)ba
Lienamine and N-[2,3-O-isopropylene
Lyden-2,3-dihydroxy-5-(1,2-
dihydroxyethyl)tetrahydrofuran-4-
Il] Valienamine Valienamine (1.0g) and 1,2:5,6-di
-O-isopropylidene-α-D-ribo-hexo
Furanose-3-ulose (2.0g) in dimethyl
Dissolved in formamide (25 ml) and 2N hydrochloric acid (0.75
ml) and sodium cyanoborohydride (1.3g)
and stir at room temperature for 40 hours. Concentrate the reaction solution under reduced pressure.
Condensed, and dimethylforma was added azeotropically with toluene.
Distill the mido. Dissolve the residue in water (100ml) and
Dowex 50W x 8 (H⁺Mold, Dow Chemical Company
(120 ml) and stirred at room temperature for 1 hour.
Ru. Add this mixture to another Dowex 50W x 8
(H⁺30 ml) on top of the column filled with
After washing the ram with water, elute with 0.5N ammonia water.
Concentrate the eluate fraction under reduced pressure and transfer to Amberlite CG-50.
(NH⁺ _Fourmold, manufactured by Rohm, & Haas, USA)
(250ml) column chromatography and elute with water.
It is separated into two components. Fraction eluted first
(210-285ml) was concentrated under reduced pressure, and
2(OH^-Mold, Dow Chemical Company, USA) (250
ml) column chromatography and elute with water. melt
After concentrating the extracted fraction under reduced pressure and lyophilizing it, N-[2,
3-O-isopropylidene-2,3-dihydroxy
C-5-(1,2-dihydroxyethyl)tetra
Hydrofuran-4-yl Valienamine white powder
(500mg) is obtained. Concentrate the fraction (290-360ml) that will be eluted later under reduced pressure.
Then, once again, Amberlight CG-50 (NH⁺ _Fourtype,
250ml) column chromatography and elute with water.
After concentrating the eluted fraction under reduced pressure and lyophilizing it, N-[2,
3,5-trihydroxy-6-hydroxymethyl
(tetrahydropyran-4-yl) valienamine
A white powder (400 mg) is obtained. eluted first
The above isopropylidene derivative obtained from the fraction
(500 mg) in water (50 ml) and dowex
50W×8(H⁺mold, 30ml) and heat at 80-90℃ for 1 hour.
Stir for a while. Add this mixture to another
50W×8(H⁺mold, 5 ml) on top of the column filled with
After washing the column with water, dissolve it with 0.5N ammonia water.
put out Concentrate the eluted fraction under reduced pressure and add Amberlite.
CG-50(NH⁺ _Fourtype, 250ml) column chromatography.
and elute with water. Concentrate the eluate fraction under reduced pressure and freeze it.
When dried, N-(2,3,5-trihydroxy-
6-hydroxymethyltetrahydropyran-4-
white powder (350 mg) of valienamine was obtained.
It will be done. N-[2,3-O-isopropylidene-2,3
-dihydroxy-5-(1,2-dihydroxye
(chill)tetrahydrofuran-4-yl]variena
Min: [α]^{twenty three} _D+172.4゜(c=1,H₂O) Elemental analysis: C₁₆H₂₇NO.₉・H₂O Calculated value (%): C, 48.48; H, 7.63; N, 3.53 Experimental value (%): C, 48.42; H, 7.47; N, 3.47 NMR(D₂O) δ: 1.55 (3H, s), 1.72 (3H,
s), 3.55 (1H, q, J=5Hz), 3.65~4.5
(10H, m), 5.12 (1H, t, J=4.5Hz),
6.09 (2H, d, J = 4.5Hz) TLC: Rf=0.58 N-(2,3,5-trihydroxy-6-hydro
roxymethyltetrahydropyran-4-yl)ba
Lienamine: [α]^{twenty three} _D+129.9゜(c=1,H₂O) Elemental analysis: C₁₃H_{twenty three}NO.₉・H₂O Calculated value (%): C, 43.94; H, 7.09; N, 3.94 Experimental value (%): C, 44.00; H, 7.31; N, 3.97 I C₅₀(maltase): 3.1×10^-FiveM I C₅₀(Satsukarase): 4.5×10^-7M TLC: Rf=0.42 Reference example 42 Methyl 4-[(1S,6S)-(4,6/5)-4,
5,6-trihydroxy-3-hydroxymethyl
-2-cyclohexen-1-yl]amino-4-
Deoxy-α-D-glucopyranoside and its
Galactopyranoside isomers Valienamine (1.5g) and methyl 6-O-a
Cetyl-2,3-O-cyclohexylidene-α-
D-xylo-hexopyranoside-4-ulose
(3.8g) dissolved in dimethylformamide (35ml)
and 2N hydrochloric acid (1.0ml) and sodium cyanoborohydride.
Add aluminum (2.0g) and stir at 60-70℃ for 15 hours.
do. The reaction solution was concentrated under reduced pressure, and the residue was diluted with 50% methanol.
Dissolved in cold water (100ml), Dowex 50W×
8(H⁺Mold, Dow Chemical Company, USA) (100ml)
Add and stir at room temperature for 1.5 hours. this mixture
In advance, use another Dowex 50W x 8 (H⁺type,
50ml) on top of the packed column, and rinse the column with water.
After washing (1), elute with 0.5N ammonia water.
The eluted fraction (0.61-1.25) was concentrated under reduced pressure and the residue
Dissolved in 4% aqueous ammonia (200ml) and stirred at room temperature.
Stir overnight. The reaction solution was concentrated to dryness under reduced pressure, and the residue
Dissolve in water (approx. 10ml) and add Amberlite CG-
50(NH⁺ _FourMold, manufactured by Rohm, & Haas, USA
country) (450ml) and eluted with water.
do. Concentrate the eluate fraction (210-325 ml) under reduced pressure and remove the residue.
Dowex 1×2 (OH^-Type, Dow Chemi
Attached to a Cal Co., USA) (850ml) column chromatograph.
When eluted with water, it is separated into two components. melt first
The resulting fraction (1.18-1.67) was concentrated under reduced pressure and then frozen.
Galactopyranoside isomer (203 mg) when dried
is obtained and then eluted fraction (2.1-3.0)
After concentrating under reduced pressure, lyophilization produces glucopyranoside.
The isomer (430 mg) is obtained. The isomer eluted first (galactopyranoside isomer
gender): [α]^{twenty two} _D+192.4゜(c=1,H₂O) Elemental analysis: C₁₄H_{twenty five}NO.₉ Calculated value (%): C, 47.86; H, 7.17; N, 3.99 Experimental value (%): C, 47.57; H, 7.15; N, 3.53 NMR(D₂O) δ: 3.42 (1H, broad d, J=
4Hz), 3.62 (3H, s), 3.6~4.5 (11H,
m), 5.04 (1H, d, J = 3.6Hz), 6.23
(1H) I C₅₀(maltase): 8.0×10^-FiveM I C₅₀(Satsukarase): 4.0×10^-6M TLC: Rf=0.40 The later eluted isomer (glucopyranoside isomer
body): [α]^{twenty two} _D+174.7゜(c=1,H₂O) Elemental analysis: C₁₄H_{twenty five}NO.₉・1/2H₂O Calculated value (%): C, 46.66; H, 7.27; N, 3.89 Experimental value (%): C, 46.26; H, 7.23; N, 3.34 NMR(D₂O) δ: 2.77-3.03 (1H, m), 3.62
(3H, s), 3.6-4.5 (11H, m), 5.02 (1H,
d, J = 3Hz), 6.11 (1H) I C₅₀(maltase): 7.2×10^-6M I C₅₀(Satsukarase): 3.2×10^-7M TLC: Rf=0.49 Reference example 43 N-(2,4-diethoxycarbonylamino-
5,6-dihydroxycyclohexyl)-7-a
Dido-7-deoxyvariolamine 7-azido-7-deoxyvariolamine
(2.0g) and N,N'-diethoxylate obtained in Reference Example 18
Cycarbonyl-4,5-O-isopropylidene-
6-oxo-2-deoxystreptamine and
N,N'-diethoxycarbonyl-5,6-O-
Isopropylidene-4-oxo-2-deoxys
A mixture of treptamine (4.0 g) was added to dimethylform.
Dissolve in Muamide (50ml) and add 2N hydrochloric acid (1.5ml).
Add sodium cyanoborohydride (2.6g),
Stir at 60-70°C for 15 hours. Concentrate the reaction solution under reduced pressure
Then, add toluene and repeat the azeotropic concentration under reduced pressure.
return. Dissolve the residue in water (200ml) and
Kusu 50W×8(H⁺Mold, manufactured by Dow Chemical Company, USA
(200 ml) and stir at 60°C for 1 hour. child
Pour the mixture into another Dowex 50W x 8 (H⁺Type, 50
ml) onto the packed column and wash the column with water.
Then, elute with 0.5N ammonia water. elution fraction
Concentrate under reduced pressure and transfer the residue to Amberlite CG-50
(NH⁺ _Fourmold, manufactured by Rohm, & Haas, USA)
(400ml) column chromatography and elute with water.
Ru. After concentrating the eluted fraction under reduced pressure and lyophilizing it, N-
(2,4-diethoxycarbonylamino-5,6
-dihydroxycyclohexyl)-7-azido-
7-deoxyvariolamine (1.6g) was obtained.
It will be done. [α]^{twenty three} _D+27.7゜(c=1,H₂O) Elemental analysis: C₁₉H₃₄N₆O_Ten・H₂O Calculated value (%): C, 43.51; H, 6.92; N, 16.02 Experimental value (%): C, 43.82; H, 6.61; N, 15.70 Reference example 44 1-N-(2,4-diethoxycarbonylamine
No-5,6-dihydroxycyclohexyl)-7
-Amino-7-deoxyvariolamine N(2,4-diethoxycarbonylamino-5,
6-dihydroxycyclohexyl)-7-azide
-7-deoxyvariolamine (1.4g) in water
(50 ml), acetic acid (1 ml) and palladium
Add Muguro (250 mg) and leave at room temperature for 4 hours in a hydrogen stream.
Stir with After removing the catalyst by filtration and washing with water, the filtrate and
Combine the washings and concentrate to dryness under reduced pressure. Residue overnight
After drying under reduced pressure in a desiccator, add to water (10ml).
Dissolve Amberlite CG-50 (NH⁺ _Fourtype, low
(manufactured by Mu, & Haas, USA) (250ml)
After washing the column with water, add 0.2N ampere.
Elute with Monia water. After concentrating the eluate fraction under reduced pressure, freeze it.
When dried, 1-N-(2,4-diethoxylic
Bonylamino-5,6-dihydroxycyclohexyl
sil)-7-amino-7-deoxyvariola
A white powder of min (850 mg) is obtained. [α]^{twenty three} _D+13.2゜(c=1,H₂O) Elemental analysis: C₁₉H₃₆N_FourO_Ten・H₂O Calculated value (%): C, 45.77; H, 7.68; N, 11.24 Experimental value (%): C, 45.88; H, 7.85; N, 10.95 Reference example 45 1.N-(2,4-diamino-5,6-dihydro
(xycyclohexyl)-7-amino-7-deoxy
Chivariolamine 1-N-(2,4-diethoxycarbonylamine
No-5,6-dihydroxycyclohexyl)-7
-amino-7-deoxyvariolamine (700
mg) in water (25 ml) and barium hydroxide.
(3.0g) and stirred at 70-80°C for 1 hour. anti
After cooling the reaction solution to room temperature, carbon dioxide gas is passed through it to produce a raw material.
The resulting precipitate is filtered off and washed with water. Combine the filtrate and washing liquid.
Amberlight CG-50 (NH⁺ _Fourtype, loam,
Column and Haas Co., USA) (100ml)
After washing the column with water, add 0.2N ammonia
Elute with aqueous ammonia and then with 0.5N ammonia water. melt
The extracted fraction was concentrated under reduced pressure and filtered with Dowex 1×2 (OH^-
Mold, Dow Chemical Co., USA) (130ml)
After washing the column with water, add 0.5N hydrochloric acid.
Elutes with Adjust the elution fraction to pH 6 and add amber
Light CG-50 (NH⁺ _Fourtype, 100ml) column chroma
After washing the column with water, wash it with 1N ammonia water.
Elute. Concentrate the eluted fraction under reduced pressure and freeze-dry it.
and 1-N-(2,4-diamino-5,6-dihyde
(roxycyclohexyl)-7-amino-7-deo
White powder (150 mg) of xyvariolamine was obtained.
It will be done. [α]^{twenty two} _D+17.7゜(c=1,H₂O) Elemental analysis: C₁₃H₂₈N_FourO₆・2H₂O Calculated value (%): C, 41.92; H, 8.66; N, 15.05 Experimental value (%): C, 41.42; H, 8.52; N, 14.84 Reference example 46 Methyl 2,3-di-O-acetyl-4-
[(1S,2S)-(2,4,5(OH)/3,5
(CH₂OH))-2,3,4-triacetoxy-5
-acetoxymethyl-5-hydroxycyclohex
sil]amino-4,6-dideoxy-α-D-g
Lucopyranoside Methyl 4-[(1S,2S)-(2,4,5
(OH)/3,5(CH₂OH))-2,3,4,5-
Tetrahydroxy-5-hydroxymethylcyclo
hexyl]amino-4,6-dideoxy-α-D
- Glucopyranoside (200mg) in pyridine (5ml)
Add acetic anhydride (3 ml) and leave at room temperature overnight.
Stir. The reaction solution was concentrated under reduced pressure, and the residue was left in the refrigerator overnight.
After drying under reduced pressure in a siccator, ethyl ether
-Add petroleum ether (1:1, approx. 30ml) overnight
When left at room temperature, hexa-O-acetyl derivatives
Crystals (265 mg) are obtained. [α]^{twenty three} _D+62.9゜(c=1, CH₃OH) Elemental analysis: C₂₆H₃₉NO.₁₅ Calculated value (%): C, 51.57; H, 6.49; N, 2.31 Experimental value (%): C, 51.68; H, 6.36; N, 2.09 NMR (CDCl)₃) δ: 1.36 (3H, d, J = 6Hz),
1.70 (1H, dd, J=3.5Hz, 15Hz), 1.97
(6H, s), 2.02 (3H, s), 2.04 (6H,
s), 2.08 (3H, s), 2.10 (1H, dd, J=
3Hz, 15Hz), 2.5~2.85 (1H, m, D₂O
When added, it changes to δ: 2.66 (t, J = 10).
), 3.37 (3H, s), 3.5~3.85 (2H, m),
3.72 (1H, d, J = 11.5Hz), 4.03 (1H,
d, J=11.5Hz), 4.7-4.9 (2H), 5.03
(1H, d, J = 10Hz), 5.04 (1H, dd, J
= 4.5Hz, 10.5Hz), 5.32 (1H, t, J = 10.5
Hz), 5.58 (1H, t, J = 10Hz) Reference example 47 Methyl 2,3,6-tri-O-acetyl-4
−[(1S,2S)−(2,4,5(OH)/3,5
(CH₂OH))-2,3,4-triacetoxy-5
-acetoxymethyl-5-hydroxycyclohex
sil]amino-4-deoxy-α-D-glucopi
Ranocide Methyl 4-[(1S,2S)-(2,4,5
(OH)/3,5(CH₂OH))-2,3,4,5-
Tetrahydroxy-5-hydroxymethylcyclo
hexyl]amino-4-deoxy-α-D-glue
Copyranoside (100 mg) was dissolved in pyridine (4 ml).
Dissolve, add acetic anhydride (2 ml) and leave at room temperature overnight.
do. Concentrate the reaction solution under reduced pressure and desiccate the residue overnight.
After drying under reduced pressure in a
Add oil ether (1:2, about 20ml) and refrigerate overnight.
If left in the refrigerator, hepta-O-acetyl derivatives
Crystals (150 mg) are obtained. [α]^{twenty three} _D+66.0゜(c=1, CH₃OH) Elemental analysis: C₂₈H₄₁NO.₁₇ Calculated value (%): C, 50.68; H, 6.23; N, 2.11 Experimental value (%): C, 50.64; H, 6.30; N, 2.21 NMR (CDCl)₃) δ: 1.73 (1H, dd, J=4Hz,
15.5Hz), 1.9-2.3 (1H), 1.97 (3H, s),
2.01 (3H, s), 2.04 (9H, s), 2.10 (3H,
s), 2.13 (3H, s), 2.75~3.2 (1H, m,
D₂When O is added, δ: 2.97 (t, J = 10Hz)
) 3.39 (3H, s), 3.5 to 3.85
(2H, m), 3.67 (1H, d, J=11.7Hz),
3.99 (1H, d, J = 11.7Hz), 4.15~4.6
(2H, m), 4.65-5.15 (4H, m), 5.34
(1H), 5.55 (1H, t, J = 10Hz) Example 1 Methyl 4-V[1S,2S)-(2,4/3,5)
-2,3,4-trihydroxy-5-hydroxy
Methylcyclohexyl]amino-4,6-dideo
xy-α-D-glucopyranoside and its 6-
Deoxygalactopyranoside isomer Validamine (2.0g) and methyl 2,3-O-
Cyclohexylidene-6-deoxy-α-D-ki
scyllo-hexopyranoside-4-ulose (methyl
2,3-O-cyclohexylidene-6-deoxy
C-α-D-xylo-4-hexopyranoseuro
(5.5g) in dimethylformamide (50ml)
Dissolved in 2N hydrochloric acid (1.5 ml) and cyanohydrogenated
Add sodium boron (2.6g) and heat at 60-70℃.
Stir for 20 hours. The reaction solution was concentrated under reduced pressure and further
Dimethylformamide is reduced azeotropically by adding ene.
Distill under pressure. Dissolve the residue in water (150ml) and
Wetx 50W x 8 (H⁺Mold, Dow Chemical Company
(150 ml) and stirred at room temperature for 2 hours.
Ru. Add this mixture to a separate dowel rack beforehand.
50W×8(H⁺30ml) on top of the column filled with
After washing the column with water (1),
Elute with aqueous ammonia. Elution fraction (0.7-1.1)
Concentrate under reduced pressure and pour the concentrated liquid (approximately 15 ml) into Amber Lie.
CG-50 (NH⁺ _FourMold, Rohm, and Haas
Co., Ltd., USA) (250 ml) and water.
Elutes with Concentrate the eluate fraction (170-250ml) under reduced pressure
Then pour the concentrated liquid (approx. 10ml) into a dowex 1x2
(OH^-Mold, Dow Chemical Company, USA) (1)
When subjected to column chromatography and eluted with water, two components
separated into The fraction eluted earlier (0.68-1.24
) and the next eluted fraction (1.25-2.74)
Concentrate each under reduced pressure. of the fraction eluted first
Pour the concentrate (approx. 20ml) into the Dowex 1x2
(OH^-Type, 850 ml) column chromatography, water
Elute with
After that, lyophilization produces 6-deoxygalactopyrano.
The side isomer (160 mg) is obtained. later eluted
Similarly, add the concentrated solution (about 20 ml) of the fraction used again.
Wex 1×2 (OH^-type, 1) column black
The eluted fraction (1.33-2.75
) is concentrated under reduced pressure and lyophilized to produce 6-deoxy
Glucopyranoside isomer (415 mg) is obtained. The isomer eluted first (6-deoxygalacto
pyranoside isomer): [α]^{twenty two} _D+162.0゜(c=1,H₂O) Elemental analysis: C₁₄H₂₇NO.₈・H₂O Calculated value (%): C, 47.31; H, 8.23; N, 3.94. Experimental value (%): C, 47.30; H, 8.35; N, 3.94. NMR(D₂O) δ: 1.2-2.4 (3H, m), 1.53
(3H, d, J = 6.5Hz), 3.08 (1H), 3.2~
4.5 (9H, m), 3.61 (3H, s), ~5.0 (1H). I C₅₀(Satsukarase): 5.6×10^-6M. TLC: Rf=0.36 The later eluted isomer (6-deoxyglucopi
Ranosidic isomer): [α]^{twenty two} _D+145.9゜(c=1,H₂O) Elemental analysis: C₁₄H₂₇NO.₈・H₂O Calculated value (%): C, 47.31; H, 8.23; N, 3.94. Experimental value (%): C, 47.37; H, 8.06; N, 4.04. NMR(D₂O) δ: 1.3-2.3 (3H, m), 1.52
(3H, d, J = 6.5Hz), 2.61 (1H, t, J
= 10Hz), 3.3 to 4.15 (9H, m), 3.62 (3H,
s), ~4.95 (1H). I C₅₀(maltase): 1.8×10^-6M I C₅₀(Satsukarase): 7.4×10^-7M TLC: Rf=0.48 Example 2 Methyl 4-[(1S,2S)-(2,4,5
(OH)/3,5(CH₂OH))-2,3,4,5-
Tetrahydroxy-5-hydroxymethylcyclo
hexyl]amino-4,6-dideoxy-α-D
-Glucopyranoside and its 6-deoxygala
Cutopyranoside isomer Variolamine (2.0g) and methyl 2,3
-O-cyclohexylidene-6-deoxy-α-
D-xylo-hexopyranoside-4-ulose
(5.3g) dissolved in dimethylformamide (50ml)
2N hydrochloric acid (1.5ml) and cyanoborohydride
Add sodium (2.6g) and stir at 65℃ for 15 hours.
do. Concentrate the reaction solution under reduced pressure and add toluene.
Dimethylformamide is distilled off under reduced pressure azeotropically.
Ru. Dissolve the residue in water (150ml) and
50W×8(H⁺(Mold, Dow Chemical Company, USA)
(150 ml) and stirred at room temperature for 2 hours. This mixture
Mix the compound in advance with another Dowex 50W x 8
(H⁺30 ml) on top of the column filled with
Wash the ram with water (1) and add 0.5N ammonia water.
Elutes with Concentrate the eluted fraction (0.65-1.33) under reduced pressure.
The concentrated liquid (approximately 20ml) is poured into Amberlite CG-
50(NH⁺ _FourMold, manufactured by Rohm, & Haas, USA
country) (250ml) and eluted with water.
do. Concentrate the eluate fraction (150-230ml) under reduced pressure and remove the residue.
Dowex 1×2 (OH^-Type, Dow Chemi
Attached to a column chromatograph (manufactured by Cal, USA) (1.5)
When eluted with water, it is separated into two components. melt first
The resulting fraction is concentrated under reduced pressure and lyophilized to give 6-
Deoxygalactopyranoside isomer (0.25g)
The obtained and then eluted fractions were concentrated under reduced pressure and then frozen.
When dried, 6-deoxyglucopyranoside isomer
body (1.47 g) is obtained. The isomer eluted first (6-deoxygalacto
pyranoside isomer): [α]^{twenty two} _D+130.7゜(c=1,H₂O) Elemental analysis: C₁₄H₂₇NO.₉・1/2H₂O Calculated value (%): C, 46.40; H, 7.79; N, 3.87. Experimental value (%): C, 46.34; H, 8.09; N, 3.51. NMR(D₂O) δ: 1.53 (3H, d, J = 6.5Hz),
1.70 (1H, dd, J=3Hz, 15.5Hz), 2.33
(1H, dd, J = 3Hz, 15.5Hz), 3.17 (1H,
dd, J=1.5Hz, 4Hz), 3.45~4.55 (9H,
m), 3.61 (3H, s), 5.0 (1H, d, J=4
Hz). I C₅₀(maltase): 6.5×10^-7M I C₅₀(Satsukarase): 2.5×10^-7M TLC: Rf=0.32 The later eluted isomer (6-deoxyglucopi
Ranosidic isomer): [α]^{twenty two} _D+105.6゜(c=1,H₂O) Elemental analysis: C₁₄H₂₇NO.₉・1/2H₂O Calculated value (%): C, 46.40; H, 7.79; N, 3.87. Experimental value (%): C, 46.64; H, 8.52; N, 3.46. NMR(D₂O) δ: 1.48 (3H, d, J = 6Hz),
1.71 (1H, dd, J=3Hz, 15.5Hz), 2.21
(1H, dd, J = 3Hz, 15.5Hz), 2.60 (1H,
t, J=9Hz), 3.37~4.35 (9H, m),
3.55 (3H, s), ~4.8 (1H). I C₅₀(maltase): 4.9×10^-9M I C₅₀(Satsukarase): 1.0×10^-8M TLC: Rf=0.44 Example 3 Methyl 4-[(1S,2S)-(2,4/3,5)-
2,3,4-trihydroxy-5-hydroxymeth
Tylcyclohexyl]amino-4-deoxy-α
-D-glucopyranoside and its galactopyranoside
nosidic isomer Validamine (2.0g) and methyl 6-O-acetate
Chil-2,3-O-cyclohexylidene-α-D
-xylo-hexopyranoside-4-ulose (meth)
Chil 6-O-acetyl-2,3-O-cyclohe
Xylidene-α-D-xylo-4-hexopyrano
-Suuroside) (4.7g) in dimethylformamide
(35ml), 2N hydrochloric acid (1.5ml) and cyano water
Add sodium borohydride (2.6g) and bring to 60-70
Stir for 18 hours at °C. Concentrate the reaction solution under reduced pressure to remove the remaining
Dissolve the substance in 50% methanol water (150ml) and
X50W×8(H⁺Mold, manufactured by Dow Chemical Company,
(USA) (150 ml) and stir at room temperature for 1.5 hours.
Add this mixture to another dowex beforehand.
50W×8(H⁺mold, 30 ml) on top of the column filled with
After washing the column with water (700ml), add 0.5N ampere.
Elute with Monia water. Elution fraction (0.7-1.5)
Concentrate to dryness under reduced pressure, and dissolve the residue in 4% aqueous ammonia.
(200ml) and leave at room temperature for 18 hours. reaction
The liquid was concentrated to dryness under reduced pressure, and the residue was dissolved in water (approximately 10 ml).
Understand, Amberlight CG-50 (NH⁺ _Fourtype, loam,
& Haas Co., USA) (450ml) Column
chromatograph and elute with water. Elution fraction (210~
305ml) under reduced pressure, and download the concentrated liquid (approximately 10ml).
X1×2 (OH^-Mold, manufactured by Dow Chemical Company,
(USA) (850ml) and dissolved in water.
fraction eluted at 0.57-0.95, 0.96-
Fractions eluted at 1.33, and 1.34 to 2.09
Separate into eluted fractions. Eluted between 1.34 and 2.09
After concentrating the fraction under reduced pressure and lyophilizing it, glucopi
A white powder (190 ml) of the ranosidic isomer is obtained.
The fraction eluted between 0.96 and 1.33 was concentrated under reduced pressure.
Pour the condensed liquid (approx. 10ml) into the dowex 1x2 again.
(OH^-270 ml) column chromatography, water
Elutes with Elute at 210-360ml with this chromatography
elutes between 0.57 and 0.95 in the first chromatogram.
The collected fractions were combined and concentrated under reduced pressure to obtain a concentrated solution (approx.
10ml) of Amberlite CG-50 (NH⁺ _FourType, 250
ml) column chromatography and elute with water. melt
The extracted fraction (150-180ml) was concentrated under reduced pressure and then lyophilized.
and white powder of galactopyranoside isomer (60
mg) is obtained. Glucopyranoside isomers: [α]^{twenty three} _D+142.0゜(c=1,H₂O) Elemental analysis: C₁₄H₂₇NO.₉・H₂O Calculated value (%): C, 45.28; H, 7.87; N, 3.77. Experimental value (%): C, 44.99; H, 8.45; N, 3.87. NMR(D₂O) δ: 1.3-2.3 (3H, m), 2.87
(1H, t, J=10Hz), 3.35~4.25 (11H,
m), 3.66 (3H, s), 5.03 (1H, d, J=
3.5Hz). I C₅₀(maltase): 4.4×10^-6M I C₅₀(Satsukaraze): 5.8×10^-7M TLC: Rf=0.43 Galactopyranoside isomers: [α]^{twenty three} _D+144.1゜(c=1,H₂O) Elemental analysis: C₁₄H₂₇NO.₉ Calculated value (%): C, 47.58; H, 7.70; N, 3.96. Experimental value (%): C, 47.53; H, 8.21; N, 4.14. I C₅₀(Satsukarase): 1.0×10^-6M TLC: Rf=0.36 Example 4 Methyl 4-[(1S,2S)-(2,4,5
(OH)/3,5(CH₂OH))-2,3,4,5-
Tetrahydroxy-5-hydroxymethylcyclo
hexyl]amino-4-deoxy-α-D-glue
Copyranoside and its galactopyranoside isomerism
body Variolamine (2.0g) and methyl 6-O
-acetyl-2,3-O-cyclohexylidene-
α-D-xylo-hexopyranoside-4-uro
(5.5g) in dimethylformamide (35ml)
Dissolve in 2N hydrochloric acid (1.5 ml) and cyanohydride.
Add sodium urethane (2.6 g) and heat at 60-70℃ for 18
Stir for an hour. Concentrate the reaction solution under reduced pressure and reduce the residue to 50%
Dissolved in % methanol water (150ml) and
50W×8(H⁺(Mold, Dow Chemical Company, USA)
(150 ml) and stir at room temperature for 1.5 hours. this
Pour the mixture in advance into another Dowex 50W
8(H⁺(type, 30 ml) on top of the column filled with
After washing the column with water (700ml), add 0.5N ammonia.
Elutes with water. Concentrate the eluted fraction (0.7-1.5) under reduced pressure.
Condensate to dryness and dissolve the residue in 4% aqueous ammonia (200ml)
Dissolve in and leave at room temperature for 15 hours. Reduce the pressure of the reaction solution
After concentration, dissolve the residue in water (approximately 10 ml) and
-Light CG-50 (NH⁺ _Fourtype, roam, and ha
(450ml) column chromatography
and elute with water. Reduce the elution fraction (250-340ml)
Concentrate under pressure and pour the concentrated liquid (approx. 10ml) into Dowex 1x.
2(OH^-Mold, Dow Chemical Company, USA) (850
ml) column chromatography, eluted with water, 0.57
Fraction eluted at ~1.14, eluted at 1.15-1.17
fractions and fractions eluted between 1.72 and 2.85
Divide into Vacuum the fraction eluted between 1.72 and 2.85
After concentration and lyophilization, glucopyranoside isomers
of white powder (375 mg) is obtained. 1.15~1.71
The fraction eluted with was concentrated under reduced pressure, and the concentrated solution (about 10
ml) and again in Dowex 1×2 (OH^-type,
270ml) column chromatography and elute with water.
In this chromatography, the fractions eluted at 250 to 550 ml and the
0.57 to 1.14 on the first Dowex 1×2 chromatograph
Combine the fractions eluted with and concentrate under reduced pressure.
Pour the condensed liquid (approx. 10ml) into Amberlite CG-50 (NH⁺ _Four
Type, 250 ml) column chromatography and elute with water.
After concentrating the eluate fraction (160-200ml) under reduced pressure, freeze it.
When dried, it forms a white powder of galactopyranoside isomer.
(160mg) is obtained. Glucopyranoside isomers: [α]^{twenty three} _D+102.1゜(c=1,H₂O) Elemental analysis: C₁₄H₂₇NO._Ten・1/2H₂O Calculated value (%): C, 44.44; H, 7.46; N, 3.70. Experimental value (%): C, 44.85; H, 7.85; N, 3.82. NMR(D₂O) δ: 1.78 (1H, dd, J=3.3Hz,
15Hz), 2.25 (1H, dd, J=3Hz, 15Hz),
2.92 (1H, t, J=10Hz), 3.55~4.2
(11H, m), 3.62 (3H, s), 5.03 (1H,
d, J = 3.8Hz). I C₅₀(maltase): 7.2×10^-8M I C₅₀(Satsukaraze): 8.0×10^-8M TLC: Rf=0.35 Galactopyranoside isomers: [α]^{twenty three} _D+105.4゜(c=1,H₂O) Elemental analysis: C₁₄H₂₇NO._Ten・H₂O Calculated value (%): C, 43.41; H, 7.55; N, 3.62. Experimental value (%): C, 43.36; H, 8.08; N, 4.01. I C₅₀(maltase): 3.2×10^-7M I C₅₀(Satsukarase): 6.2×10^-7M TLC: Rf=0.33 Example 5 [(1S, 2S) - (2, 4/3, 5) - 2, 3, 4
-trihydroxy-5-hydroxymethylcyclo
hexyl] [(2S)-(2,6/3,4)-4-ben
Zyloxycarbonylamino-2,3-dihydro
xy-6-hydroxymethylcyclohexyl]a
min Validamine (1.5g) and (2R)-(2,6/3,
4)-1,3-diacetoxy-4-benzyloxy
Cycarbonylamino-6-trityloxymethyl
Add cyclohexanone (5.4 g) to dimethylforma.
(35 ml), 2N hydrochloric acid (1.15 ml) and syringe.
Add sodium anoborohydride (2.0g),
Stir at 60-70°C for 15 hours. Concentrate the reaction solution under reduced pressure
Add the residue to a mixture of water and ethyl acetate, add vinegar
Separate the ethyl acetate layer and extract the aqueous layer with ethyl acetate.
Ru. The ethyl acetate extract was collected, washed with water, and diluted with sodium sulfate.
After drying with aluminum, concentrate under reduced pressure. Ethyl in the residue
Add ether and dilute the resulting syrup with 10% ammonia.
Water-ethanol (1:2) mixture (300ml)
and stir overnight at room temperature. Concentrate the reaction solution under reduced pressure.
The residue was dissolved in 80% acetic acid water (200 ml),
Stir at 80°C for 2 hours. Concentrate the reaction solution under reduced pressure and remove the residue.
Add the distillate to a mixture of water and ethyl acetate and separate the aqueous layer.
Release, Dowex 50W x 8 (H⁺Type, Dow Chemi
Attached to a column chromatograph (manufactured by Cal, USA) (150ml)
vinegar. After washing the column with water, elute with 0.5N ammonia water.
After concentrating the eluate fraction under reduced pressure, add the concentrated solution (20 ml).
Amberlight CG-50 (NH⁺ _Fourtype, roam, ann
De Haas Co., USA) (250ml) Column Chroma
eluted with water. Concentrate the eluted fraction under reduced pressure
After that, when freeze-dried [(1S, 2S) - (2, 4/3,
5)-2,3,4-trihydroxy-5-hydro
oxymethylcyclohexyl] [(2S)-(2,6/
3,4)-4-benzyloxycarbonylamino
-2,3-dihydroxy-6-hydroxymethyl
cyclohexyl]amine (980 ml) is obtained. [α]^23D+37.4° (c=0.5, H₂O) Elemental analysis: C_{twenty two}H₃₄N₂O₉・1/2H₂O Calculated value (%): C, 55.10; H, 7.36; N, 5.84. Experimental value (%): C, 55.15; H, 7.35; N, 5.71. I C₅₀(Satsukaraze): 7.0×10^-7M TLC: Rf=0.65 Example 6 [(1S, 2S) - (2, 4/3, 5) - 2, 3, 4
-trihydroxy-5-hydroxymethylcyclo
hexyl] [(1S, 2S)-(2,6/3,4)-4-
Amino-2,3-dihydroxy-6-hydroxy
Methyl]amine and [(1S,2S)-(2,4/
3,5)-2,3,4-trihydroxy-5-hyde
Droxymethylcyclohexyl] [(1R, 2S)-
(2,6/3,4)-4-amino-2,3-dihyde
roxy-6-hydroxymethyl]amine [(1S, 2S) - (2, 4/3, 5) - 2, 3, 4
-trihydroxy-5-hydroxymethylcyclo
hexyl] [(2S)-(2,6/3,4)-4-ben
Zyloxycarbonylamino-2,3-dihydro
xy-6-hydroxymethylcyclohexyl]a
Min (880 mg) was dissolved in water (150 ml), and acetic acid (1
ml) and palladium black (300 mg) and hydrogen stream
Stir at medium room temperature for 4 hours. After reaction, remove catalyst by filtration
After washing with water, combine the filtrate and washing liquid and concentrate to dryness under reduced pressure.
Ru. Dissolve the residue in water (approx. 20ml) and add amber
Ito CG-50 (NH⁺ _FourType, Rohm, and Haas
Co., Ltd., USA) (250ml) column chromatography.
After washing the column with water and eluting with 0.1N ammonia water,
It is separated into two components and eluted. Fraction eluted earlier (eluted between 1.45 and 1.90)
After concentrating the fraction under reduced pressure and freeze-drying, it becomes a white powder.
(80 mg) was obtained and then the eluted fraction (2.0 mg) was obtained.
The fraction eluted at ~2.9) was concentrated under reduced pressure and then frozen.
When dried, a white powder (370 mg) is obtained. Isomers eluted first: [α]^{twenty two} _D+66.4゜(c=0.5,H₂O) Elemental analysis: C₁₄H₂₈N₂O₇・H₂O Calculated value (%): C, 47.44; H, 8.53; N, 7.91. Experimental value (%): C, 47.38; H, 8.11; N, 7.68. I C₅₀(maltase): 2.9×10^-6M I C₅₀(Satsukalase): 1.6×10^-6M Later eluted isomers: [α]^{twenty two} _D+24.5° (c=1.0, H₂O) Elemental analysis: C₁₄H₂₈N₂O₇・H₂O Calculated value (%): C, 47.44; H, 8.53; N, 7.91. Experimental value (%): C, 47.52; H, 8.38; N, 7.61. I C₅₀(Satsukarase): 7.8×10^-FiveM Example 7 [(1S,2S)-(2,4,5(OH)/3,5
(CH₂OH))-2,3,4,5-tetrahydroxy
C-5-hydroxymethylcyclohexyl]
[(1S,2S)-(2,6/3,4)-4-benzyl
oxycarbonylamino-2,3-dihydroxy-
6-hydroxymethylcyclohexyl amine
and [(1S,2S)-(2,4,5(OH)/3,5
(CH₂OH)-2,3,4,5-tetrahydroxy
-5-hydroxymethylcyclohexyl] [(1R,
2S)-(2,6/3,4)-4-benzyloxyca
Rubonylamino-2,3-dihydroxy-6-hyperoxygenate
Droxymethylcyclohexylamine Variolamine (2.0g) and (2R)-(2,
6/3,4)-1,3-diacetoxy-4-ben
Zyloxycarbonylamino-6-trityloxy
Dimethylcyclohexanone (7.3g) in dimethyl
Dissolved in formamide (50 ml) and 2N hydrochloric acid (1.5
ml) and sodium cyanoborohydride (2.6g)
and stir at 60-70°C for 17 hours. Reduce reaction solution
Concentrate under pressure and add the residue to a mixture of water and ethyl acetate.
Separate the ethyl acetate layer and add the aqueous layer to vinegar.
Extract with ethyl acid. Combine the ethyl acetate extracts.
After washing with water and drying with sodium sulfate, concentrate under reduced pressure.
do. Residue 10% ammonia water-ethanol
(1:2) mixture (300ml) and leave it at room temperature.
Stir at night. The reaction solution was concentrated to dryness under reduced pressure, and the residue was
Dissolve in 80% acetic acid water (200ml) and stir at 80℃ for 2 hours.
Stir. The reaction solution was concentrated under reduced pressure, and the residue was mixed with water and acetic acid.
Add to the mixture with ethyl, distribute, and separate the aqueous layer.
Dowex 50W x 8 (H⁺Type, Dow Chemica
Column chromatography (manufactured by Le Co., Ltd., USA) (150 ml)
After washing with water, elute with 0.5N ammonia water. melt
Concentrate the extracted fraction under reduced pressure, and transfer the concentrated liquid (approximately 15 ml) to the amba
-Light CG-50 (NH⁺ _Fourtype, roam, and ha
(250ml) column chromatography
When eluted with water, it is separated into two components. That's it
After concentrating the eluate fraction under reduced pressure and lyophilizing it, the
White powder from the fraction eluted in (90-290ml)
(980 mg) in the fraction eluted later (300-780 ml)
A white powder (305 mg) is obtained. Isomers eluted first: [α]^{twenty three} _D+3.4゜(c=0.5,H₂O) Elemental analysis: C_{twenty two}H₃₄N₂O_Ten・H₂O Calculated value (%): C, 52.37; H, 7.19; N, 5.55. Experimental value (%): C, 52.34; H, 6.98; N, 5.73. TLC: Rf=0.62 Isomers eluted later: [α]^{twenty three} _D+8.2゜(c=0.5,H₂O) Elemental analysis: C_{twenty two}H₃₄N₂O_Ten・H₂O Calculated value (%): C, 52.37; H, 7.19; N, 5.55. Experimental value (%): C, 52.49; H, 7.00; N, 5.52. TLC: Rf=0.64 Example 8 [(1S,2S)-(2,4,5(OH)/3,5
(CH₂OH))-2,3,4,5-tetrahydroxy
C-5-hydroxymethylcyclohexyl]
[(1S,2S)-(2,6/3,4)-4-amino-
2,3-dihydroxy-6-hydroxymethylsilane
Chlorhexyl]amine and [(1S,2S)-(2,
4,5(OH)/3,5(CH₂OH)-2,3,4,
5-tetrahydroxy-5-hydroxymethylsil
Chlorhexyl] [(1R,2S)-(2,6/3,4)-
4-amino-2,3-dihydroxy-6-hydro
oxymethylcyclohexylamine 4-benzyloxycarbo obtained in Example 7
Among Nylamino derivatives, Amberlite CG-50
Components eluted first in column chromatography (980
mg) in water (150 ml), acetic acid (0.4 ml) and
Add palladium black (350 mg) and add
Stir at room temperature for an hour. After the reaction, remove the catalyst by filtration and add water.
After washing, the filtrate and washing liquid are combined and concentrated to dryness under reduced pressure. Residue
Dissolve the distillate in water (about 10ml) and add Amberlite.
CG-50(NH⁺ _FourMold, manufactured by Rohm, & Haas.
Column chromatography (USA) (250ml)
After washing with water, elute with 0.1N aqueous ammonia. Elution
The fractions were concentrated under reduced pressure and filtered with Dowex 1×2 (OH^-
Mold, Dow Chemical Co., USA) (250ml)
Apply to mucochromatography and elute with water. Reduce elution fraction
After pressure concentration and lyophilization, the corresponding 4-amino derivative
A white conductor powder (340 mg) is obtained. [α]^{twenty two} _D-9.4゜(c=1,H₂O) Elemental analysis: C₁₄H₂₈N₂O₈・3/2H₂O Calculated value (%): C, 44.32; H, 8.24; N, 7.38. Experimental value (%): C, 44.25; H, 8.45; N, 7.18. I C₅₀(maltase): 3.3×10^-6M I C₅₀(Invertase): 2.5×10^-7M Another 4-benzyl obtained in Example 7
Oxycarbonylamino derivative (Amberlite)
Products eluted later in column chromatography on CG-50
) (110 mg) in water (50 ml), acetic acid (0.1
ml) and palladium black (35 mg) and hydrogen gas
Stir at room temperature for 4 hours under running water. After reaction, remove catalyst by filtration
After washing with water, combine the filtrate and washing liquid and concentrate to dryness under reduced pressure.
Ru. Dissolve the residue in water (approx. 10ml) and add amber
Ito CG-50 (NH⁺ _FourType, Rohm, and Haas
Co., Ltd., USA) (130ml) column chromatography.
After washing the column with water, elute with 0.1N ammonia water.
Corresponding results can be obtained by concentrating the eluted fraction under reduced pressure and then freeze-drying it.
A white powder (54 mg) of 4-amino derivative was obtained.
Ru. [α]^{twenty two} _D+22.4゜(c=0.5,H₂O) Elemental analysis: C₁₄H₂₈N₂O₈・2H₂O Calculated value (%): C, 43.29; H, 8.30; N, 7.21. Experimental value (%): C, 43.48; H, 8.35; N, 6.81. I C₅₀(maltase): 2.8×10^-6M I C₅₀(Satsukarase): 2.3×10^-7M Example 9 N-(2,3,5-trihydroxy-6-hydro
roxymethyltetrahydropyran-4-yl)ba
Lyolamine and N-[2,3-O-isopropylene
Lopylidene-2,3-dihydroxy-5-(1,
2-dihydroxyethyl)tetrahydrofuran-
4-yl]variolamine Variolamine (4.0g) and 1,2:5,6
-di-O-isopropylidene-α-D-ribo-he
Add xofuranose-3-ulose (8.0g)
Dissolved in chillformamide (100ml) and 2N hydrochloric acid
(3 ml) and sodium cyanoborohydride (5.2
g) and stir at 60-70°C for 15 hours. reaction solution
was concentrated under reduced pressure, and dimethyl was further azeotroped with toluene.
Formamide is distilled off under reduced pressure. Dilute the residue with water (1
) and dowex 50W x 8 (H⁺type, da
Add U. Chemical Co., USA) (400 ml) and bring to room temperature.
Stir for 1 hour. Add this mixture in advance to
Another dowex 50W x 8 (H⁺Fill mold, 100ml).
Add on top of the packed column, and after washing the column with water,
Elute with 0.5N ammonia water. Evacuate the elution fraction
Concentrate and convert the residue to Amberlite CG-50 (NH⁺ _Four
Model, Rohm, & Haas, USA) (1.1
) and eluted with water, 2
separated into components. Concentrate the fraction eluted first (440-720ml) under reduced pressure.
and remove the residue by Dowex 1×2 (OH^-type, da
U Chemical Co., USA) (550ml) column chromatography
eluted with water. Concentrate the eluted fraction under reduced pressure
After that, freeze-drying produces N-[2,3-O-isopropylene
Pylidene-2,3-dihydroxy-5-(1,2
-dihydroxyethyl)tetrahydrofuran-4
-yl]variolamine white powder (2.8g)
is obtained. The fraction eluted later (780-1050ml) was concentrated under reduced pressure.
Shrink and remove the residue again with Amberlite CG-50
(NH⁺ _Four500ml) column chromatography, water
Elutes with After concentrating the eluate fraction under reduced pressure, freeze-dry it.
and N-[2,3,5-trihydroxy-6-hypolymer
(Droxymethyltetrahydropyran-4-yl)
A white powder (1.05g) of variolamine was obtained.
Ru. The above isoprop obtained from the fraction eluted earlier
Lopylidene derivative (450mg) dissolved in water (50ml)
Dowex 50W x 8 (H⁺mold, 30ml)
and stir at 80-90°C for 1 hour. Add this mixture to
In advance, another Dowex 50W x 8 (H⁺Type, 5
ml) onto the packed column and wash the column with water.
Then, elute with 0.5N ammonia water. elution fraction
Concentrate under reduced pressure and transfer the residue to Amberlite CG-50
(NH⁺ _Four250 ml) column chromatography, water
It elutes with After concentrating the eluate fraction under reduced pressure, freeze-dry it.
and N-(2,3,5-trihydroxy-6-hydroxy
(Droxymethyltetrahydropyran-4-yl)
A white powder (350 mg) of variolamine was obtained.
Ru. N-[2,3-O-isopropylidene-2,3
-dihydroxy-5-(1,2-dihydroxye
(chill)tetrahydrofuran-4-yl]variau
Luamine: [α]^{twenty three} _D+72.4゜(c=1,H₂O) Elemental analysis: C₁₆H₂₉NO._Ten・H₂O Calculated value (%): C, 46.48; H, 7.56; N, 3.39. Experimental value (%): C, 46.71; H, 7.22; N, 3.47. NMR(D₂O) δ: 1.51 (3H, s), 1.67 (1H,
dd, J = 3.5Hz, 15Hz), 2.27 (1H, dd, J
=3Hz, 15Hz), 3.35~4.2 (11H, m),
5.06 (1H, t, J=4Hz), 6.04 (1H, d,
J=4Hz). I C₅₀(Satsukarase): 3.7×10^-FiveM TLC: Rf=0.44 N-(2,3,5-trihydroxy-6-hydro
roxymethyltetrahydropyran-4-yl)ba
Liolamine: [α]^{twenty three} _D+31.3゜(c=1,H₂O) Elemental analysis: C₁₃H_{twenty five}NO._Ten・1/2H₂O Calculated value (%): C, 42.85; H, 7.19; N, 3.84. Experimental value (%): C, 42.56; H, 7.11; N, 3.84. I C₅₀(maltase): 1.1×10^-7M I C₅₀(Satsukarase): 9.0×10^-8M TLC: Rf=0.31 Example 10 N-(2,3,5-trihydroxy-6-hydro
roxymethyltetrahydropyran-4-yl)ba
Lydamine and N-[2,3-O-isopropylene
Den-2,3-dihydroxy-5-(1,2-di
hydroxyethyl)tetrahydrofuran-4-y
[Le] Validamine Validamine 2.0g and 1,2:5,6-di-O-
Isopropylidene-α-D-ribo-hexofurano
-3-ulose (4.0g) in dimethylform
Dissolved in amide (50 ml) and mixed with 2N hydrochloric acid (1.5 ml).
Add sodium anoborohydride (2.6g),
Stir at room temperature for 60 hours. Concentrate the reaction solution under reduced pressure and
Reduce dimethylformamide azeotropically with toluene to
Distill under pressure. Dissolve the residue in water (200ml) and
Wetx 50W x 8 (H⁺Mold, Dow Chemical Company
(200 ml) and stirred at room temperature for 1 hour.
Ru. Add this mixture to a separate dowel rack beforehand.
50W×8(H⁺type, 100ml) of a column packed with
After washing the column with water, add 0.5N ammonia water.
Elutes with Concentrate the eluate fraction under reduced pressure and remove the residue.
Amberlight CG-50 (NH⁺ _Fourtype, roam, ann
De Haas Co., USA) (400ml) Column Chroma
When it is eluted with water, it is separated into two components. Concentrate the first eluted fraction (210-285ml) under reduced pressure.
and remove the residue by Dowex 1×2 (OH^-type, da
U Chemical Co., USA) (450ml) Column Chromatin
eluted with water. Concentrate the eluted fraction under reduced pressure
After that, freeze-drying produces N-[2,3-O-isopropylene
Pylidene-2,3-dihydroxy-5-(1,2
-dihydroxyethyl)tetrahydrofuran-4
White powder (850 mg) of Validamine was obtained.
It will be done. Concentrate the fraction (290-400ml) that will be eluted later under reduced pressure.
and remove the residue with Amberlite CG-50 again.
(NH⁺ _Fourtype, 400 ml) column chromatography, water
Elutes with After concentrating the eluate fraction under reduced pressure, freeze-dry it.
and N-(2,3,5-trihydroxy-6-hydroxy
(Droxymethyltetrahydropyran-4-yl)
A white powder (500 mg) of Validamine is obtained. N-[2,3-O-isopropylidene-2,3
-dihydroxy-5-(1,2-dihydroxye
(chill)tetrahydrofuran-4-yl]validami
hmm: [α]^{twenty three} _D+120.8゜(c=1,H₂O) Elemental analysis: C₁₆H₂₉NO.₉・1/2H₂O Calculated value (%): C, 49.47; H, 7.79; N, 3.61. Experimental value (%): C, 49.52; H, 7.72; N, 3.71. NMR(D₂O) δ: 1.2-2.35 (3H, m), 1.53
(3H, s), 1.70 (3H, s), 3.25~4.3
(11H, m), 5.06 (1H, t, J=4Hz),
6.05 (1H, d, J=4Hz). TLC: Rf=0.55 N-(2,3,5-trihydroxy-6-hydro
Roxymethyltetrahydropyran-4-yl]
Ridamine: [α]²⁸ _D+82.1゜(c=1,H₂O) Elemental analysis: C₁₃H_{twenty five}NO.₉・1/2H₂O Calculated value (%): C, 44.82; H, 7.52; N, 4.02. Experimental value (%): C, 44.92; H, 7.89; N, 4.13. I C₅₀(maltase): 1.9×10^-6M I C₅₀(Satsukarase): 1.7×10^-7M TLC: Rf=0.36 Example 11 N-(2,4-diethoxycarbonylamino-
5,6-dihydroxycyclohexyl) vario
Ruamine Obtained with variolamine (2.0g) and Reference Example 18
N,N'-diethoxycarbonyl-4,5-
O-isopropylidene-6-oxo-2-deoxy
Cystreptamine and N,N'-diethoxyca
rubonyl-5,6-O-isopropylidene-4-
Mixture of oxo-2-deoxystreptamine
(4.0g) dissolved in dimethylformamide (50ml)
and 2N hydrochloric acid (1.2ml) and sodium cyanoborohydride.
Add aluminum (2.6g) and stir at 60-70℃ for 15 hours.
do. Concentrate the reaction solution under reduced pressure, add toluene and share.
Dimethylformamide is distilled off under reduced pressure while boiling. Residue
Dissolve the distillate in water (200 ml) and use Dowex 50W.
×8(H⁺Mold, Dow Chemical Company, USA) (200
ml) and stir at 60°C for 1 hour. this mixture
In advance, use another Dowex 50W x 8 (H⁺
50 ml) on top of the column filled with
After washing with water, elute with 0.5N ammonia water. Elution fraction
Concentrate the fraction under reduced pressure, and remove the residue from Amberlite CG-
50(NH⁺ _FourMold, manufactured by Rohm, & Haas, USA
country) (400ml) and eluted with water.
do. After concentrating the eluted fraction under reduced pressure and lyophilizing it, N
-(2,4-diethoxycarbonylamino-5,
6-dihydroxycyclohexyl)variola
A white powder of min (1.75 g) is obtained. [α]^{twenty two} _D+14.7゜(c=1,H₂O) Elemental analysis: C₁₉H₃₅N₃O₁₁・H₂O Calculated value (%): C, 45.68; H, 7.47; N, 8.41. Experimental value (%): C, 45.59; H, 7.41; N, 8.29. I C₅₀(maltase): 9.0×10^-6M I C₅₀(Satsukarase): 4.9×10^-6M Example 12 N-(2,4-diamino-5,6-dihydroxy
cyclohexyl)variolamine N-(2,4-diethoxycarbonylamino-
5,6-dihydroxycyclohexyl) vario
Dissolve Ruamine (1.5g) in water (50ml) and
Add barium chloride (6g) and stir at 70-80℃ for 4 hours.
Stir. After cooling the reaction solution to room temperature, carbon dioxide gas
The precipitate formed is filtered off and washed with water. With filtrate
Combine the washing liquid and Amberlite CG-50 (NH⁺ _Fourtype,
Manufactured by Rohm, & Haas, USA) (250ml)
After applying column chromatography and washing the column with water, 0.5N
Elute with aqueous ammonia. Concentrate the eluted fraction under reduced pressure
and the residue was treated with Amberlite CG-50 (NH⁺ _Fourtype,
100ml) column chromatography and wash the column with water.
Then, when eluted with 0.1N ammonia water, it separates into 3 components.
be separated. After concentrating the first eluted fraction under reduced pressure,
When freeze-dried, it becomes a white powder (43 mg) (abbreviated as isomer A).
) is obtained. Similarly for the next eluted fraction.
After concentration under reduced pressure and lyophilization, it becomes a white powder (100mg)
(abbreviated as isomer B) is obtained. last eluted
The fractions were concentrated under reduced pressure, and the residue was filtered 1x by Dowex.
2(OH^-Mold, Dow Chemical Company, USA) (180
ml) column chromatography and elute with water.
After concentrating the fraction under reduced pressure and lyophilizing it, a white powder (40%
mg) (abbreviated as isomer C) is obtained. more columns
After eluting with 0.5N hydrochloric acid and adjusting the eluted fraction to pH 6,
Amberlight CG-50 (NH⁺ _Fourmold, 100ml)
After washing the column with water, add 0.3N ampere.
Elute with Monia water. After concentrating the eluate fraction under reduced pressure, freeze it.
When dried, it becomes a white powder (160 mg) (abbreviated as isomer D).
) is obtained. Isomer D: [α]^{twenty two} _D+26.6゜(c=1,H₂O) Elemental analysis: C₁₃H₂₇N₃O₇・3/2H₂O Calculated value (%): C, 42.85; H, 8.30; N,
11.53. Experimental value (%): C, 42.83; H, 8.48; N,
11.24. I C₅₀(maltase): 1.0×10^-6M I C₅₀(Satsukarase): 8.8×10^-8M Example 13 N-(2,4-diethoxycarbonylamino-
5,6-dihydroxycyclohexyl)validami
hmm Validamine (2.0g) and N obtained in Reference Example 18,
N'-diethoxycarbonyl-4,5-O-iso
Propylidene-6-oxo-2-deoxystre
Putamine and N,N'-diethoxycarbonyl
-5,6-O-isopropylidene-4-oxo-
2-deoxystreptamine mixture (4.0g)
Dissolved in dimethylformamide (50ml) and diluted with 2N
Hydrochloric acid (1.5ml) and sodium cyanoborohydride
(2.6 g) and stirred at 60-70°C for 22 hours. anti
The reaction solution was concentrated under reduced pressure, and toluene was added to azeotropically.
Dimethylformamide is distilled off under reduced pressure. residue
Dissolve in water (200 ml) and use Dowex 50W x 8
(H⁺mold, manufactured by Dow Chemical Company, USA) (200ml)
Add and stir at 60°C for 1 hour. Add this mixture to
In advance, another Dowex 50W x 8 (H⁺Type, 50
ml) on top of the packed column and wash the column with water.
Then, elute with 0.5N ammonia water. elution fraction
Concentrate under reduced pressure and transfer the residue to Amberlite CG-50
(NH⁺ _Fourmold, manufactured by Rohm, & Haas, USA)
(400ml) column chromatography and elute with water.
Ru. After concentrating the eluted fraction under reduced pressure and lyophilizing it, N-
(2,4-diethoxycarbonylamino-5,6
-dihydroxycyclohexyl)validamine white
A colored powder (1.1 g) is obtained. [α]^{twenty three} _D+39.9゜(c=1,H₂O) Elemental analysis: C₁₉H₃₅N₃O_Ten・H₂O Calculated value (%): C, 47.19; H, 7.71; N, 8.69. Experimental value (%): C, 46.64; H, 7.38; N, 8.60. Example 14 N-(2,4-diamino-5,6-dihydroxy
cyclohexyl)validamine N-(2,4-diethoxycarbonylamino-
5,6-dihydroxycyclohexyl)validami
(900 mg) in water (35 ml) and
(4.0g) and stir at 70-80℃ for 7.5 hours.
Ru. Cool the reaction solution to room temperature and pass carbon dioxide gas through it.
Then, the precipitate formed is filtered off and washed with water. Filtrate and washing liquid
and Amberlight CG-50 (NH⁺ _Fourtype, lo
(manufactured by Ham, & Haas, USA) (150ml)
After applying ram chromatography and washing the column with water, add 0.1N
Elute with aqueous ammonia. Concentrate the eluted fraction under reduced pressure,
Dowex 1×2 (OH^-type, dau ke
Column chromatography manufactured by Michal (USA) (120ml)
After washing the column with water, elute with 0.5N hydrochloric acid.
Adjust the elution fraction to pH 6 and add Amberlite CG-
50(NH⁺ _Fourtype, 100ml) column chromatography,
After washing the column with water, elute with 0.3N aqueous ammonia.
After concentrating the eluted fraction under reduced pressure and lyophilizing it, N-
(2,4-diamino-5,6-dihydroxysic
Lohexyl) Validamine white powder (100mg)
can get. [α]^{twenty two} _D+52.4゜(c=0.5, H₂O) Elemental analysis: C₁₃H₂₇N₃O₆・2H₂O Calculated value (%): C, 43.68; H, 8.74; N,
11.76. Experimental value (%): C, 43.58; H, 8.81; N,
11.90. I C₅₀(Satsukarase): 8.8×10^-7M

Claims (1)

[Claims] 1. General formula [In the formula, A is a hydrogen atom or a hydroxyl group; X ¹ is an oxygen atom, and ^{X 2 is} a group represented by hydroxymethyl group) and X ² is an oxygen atom, or X ¹ is a group represented by the formula (Z is a hydrogen atom) and X ² is a (indicates an amino group that may be protected with an aralkyloxycarbonyl group), or
Alternatively, X ¹ and X ² together represent an oxygen atom; Y represents a lower alkyl group which may have a hydroxyl group, an amino group which may be protected with a hydroxyl group or an alkoxycarbonyl group. However, A is a hydroxyl group,
When X ¹ is -CH ₂ - and X ² is [Formula] (where Z represents a hydroxyl group or an amino group), Y is not a methyl group. ] A compound represented by. 2 General formula Compounds represented by [wherein A represents a hydrogen atom or a hydroxyl group] and the general formula [In the formula, ^{X 1} ′ is an oxygen atom and X ² ′ is a group represented by [Formula] (Z′ is a lower alkoxy group), or ) in which X ² ′ represents an oxygen atom, or X ¹ ′ represents a group represented by [Formula] (Z′ is a hydrogen atom) in which X ² ′ represents or an amino group which may be protected with a group or an aralkyloxycarbonyl group, or X ¹ ′ and X ² ′ together represent an oxygen atom; Y′ has a hydroxyl group; Indicates an amino group that is optionally protected with a lower alkyl group, a hydroxyl group, or an alkoxycarbonyl group. However, when A is a hydroxyl group, X ¹ ' is -CH ₂ -, and X ² ' is [Formula] (where Z' represents a protected amino group),
Y is not a methyl group. A general formula characterized by reacting a cyclic ketone represented by [the hydroxyl group in this compound may be protected], followed by a reduction reaction, and optionally a deprotection reaction. [In the formula, A has the same meaning as above; X ¹ is an oxygen atom and X ² is a group represented by the formula (Z represents a lower alkoxy group ⁾ , or hydroxymethyl group) in which X ² represents an oxygen atom, or X ¹ represents a group represented by [Formula] (Z represents a hydrogen atom and X ² represents a (indicates an amino group which may be protected with an oxycarbonyl group), or X ¹ , X ²
together represent an oxygen atom; Y represents a lower alkyl group which may have a hydroxyl group, an amino group which may be protected with a hydroxyl group or an alkoxycarbonyl group. However, A is a hydroxyl group, X ¹ is -CH ₂ -,
When X ² is [Formula] (where Z represents a hydroxyl group or an amino group), Y is not a methyl group. ] A method for producing a compound represented by. 3 General formula [In the formula, A is a hydrogen atom or a hydroxyl group; X ¹ is an oxygen atom, and ^{X 2 is} a group represented by hydroxymethyl group) and X ² is an oxygen atom, or X ¹ is a group represented by the formula (Z is a hydrogen atom) and X ² is a (indicates an amino group that may be protected with an aralkyloxycarbonyl group), or
Alternatively, X ¹ and X ² together represent an oxygen atom; Y represents a lower alkyl group which may have a hydroxyl group, an amino group which may be protected with a hydroxyl group or an alkoxycarbonyl group. However, A is a hydroxyl group,
When X ¹ is -CH ₂ - and X ² is [Formula] (where Z represents a hydroxyl group or an amino group), Y is not a methyl group. α- containing the compound represented by ]
Glucosidase inhibitor.