HK1167855A

HK1167855A - Novel substituted indazole and aza-indazole derivatives as gamma secretase modulators

Info

Publication number: HK1167855A
Application number: HK12108465.5A
Authority: HK
Inventors: François Paul BISCHOFF; Henricus Jacobus Maria Gijsen; Serge Maria Aloysius Pieters; Garrett Berlond Minne
Original assignee: Janssen Pharmaceuticals, Inc.
Priority date: 2009-05-07
Filing date: 2010-05-05
Publication date: 2012-12-14

Description

Novel substituted indazole and aza-indazole derivatives as gamma-secretase modulators

Technical Field

The present invention relates to novel substituted indazole and aza-indazole derivatives useful as gamma secretase modulators. The invention also relates to processes for the preparation of such novel compounds, pharmaceutical compositions comprising said compounds as active ingredient and the use of said compounds as medicaments.

Background

Alzheimer's Disease (AD) is a progressive neurodegenerative disease characterized by loss of memory, recognition and behavioral stability. AD afflicts 6-10% of people over 65 years of age and up to 50% of people over 85 years of age. This is the major cause of dementia and the third leading cause of death after cardiovascular disease and cancer only. There is currently no effective treatment for AD. The total net cost of AD in the united states is over one billion dollars per year.

AD does not have a simple etiology, however, it is associated with certain risk factors including (1) age, (2) family history, and (3) head trauma; other factors include environmental toxins and low education. Specific neuropathological lesions in the limbus and cerebral cortex include intracellular neurofibrillary tangles composed of phosphorylated tau protein and extracellular deposits of amyloid beta peptide fibrils aggregates (amyloid plaques). The major components of amyloid plaques are amyloid beta (a- β, Abeta or a β) peptides of varying lengths. One such variant is the A β 1-42-peptide (Abeta-42), which is believed to be the major causative agent of amyloid formation. Another variant is the A.beta.1-40-peptide (Abeta-40). Amyloid beta protein is the proteolytic product of a precursor protein, beta amyloid precursor protein (beta-APP or APP).

The family of AD, the early-onset autosomal dominant forms of inheritance, are associated with missense mutations in the beta-amyloid precursor protein (beta-APP or APP) and in the presenilin proteins 1 and 2. In some patients, late-onset forms of AD are associated with specific alleles of the apolipoprotein e (apoe) gene, and with recent findings of mutations in α 2-macroglobulin, which may be associated with at least 30% of the AD population. Despite this heterogeneity, all forms of AD show similar pathological consequences. Genetic analysis provides the best clues for a logical approach to the treatment of AD. All mutations found to date affect the quantity or quality of production of amyloid peptides called Abeta-peptides (a β), particularly α β 42 (amyloidogenic peptides), and provide strong support for the "amyloid cascade hypothesis" of AD (Tanzi and Bertram, 2005, Cell 120, 545). The possible link between a β peptide production and AD pathology emphasizes the need for a better understanding of the mechanisms of a β production and strongly warrants a therapeutic approach in regulating a β levels.

The release of a β peptides is regulated by at least two proteolytic activities, referred to as β -and γ -secretase cleavage at the N-terminus (Met-Asp bond) and C-terminus (residues 37-42) of a β peptides, respectively. In the secretory pathway, there is evidence that β -secretase cleavage first results in secretion of s-APP β (s β) and retention of the 11kDa membrane-bound carboxy-terminal fragment (CFT). The latter is believed to produce a β peptide upon gamma-secretase cleavage. The amount of the longer isoform, a β 42, is selectively increased in patients with certain mutations in a particular protein (presenilin), and these mutations are associated with early-onset familial alzheimer's disease. Thus, a β 42 is believed by many researchers to be the major cause of the pathogenesis of alzheimer's disease.

It is now clear that gamma-secretase activity cannot be attributed to a single protein, and is in fact associated with the assembly of different proteins.

The γ (γ) -secretase activity is located in a multiplex protein complex containing at least four components: presenilin (PS) heterodimer, apo-1, and pen-2. PS heterodimers are amino-and carboxy-terminal PS fragments generated from precursor proteins via endoproteolysis. The two aspartates of the catalyst site are located at the interface of this heterodimer. Dumb proteins have recently been proposed to act as gamma-secretase-matrix receptors. The functions of the other members of the gamma-secretase are not known, but all are required for activity (Steiner, 2004.Curr. Alzheimer Research 1 (3): 175-.

Accordingly, while the molecular mechanism of the second cleavage step has remained elusive to date, the γ -secretase-complex has become one of the primary objectives in finding compounds for the treatment of alzheimer's disease.

Various Strategies have been proposed for the gamma-secretase enzyme of Alzheimer's disease, including the development of inhibitors of substrate specificity and modulators of gamma-secretase activity directed at the catalytic site (Marjaux et al, 2004. Current Drug development: Therapeutic Strategies (Drug Discovery: Therapeutic Strategies), Vol.1, 1-6). Thus, a number of compounds targeting Secretases (Larner, 2004. Secretases as therapeutic targets in Alzheimer's disease patients) 2000-2004.Expert Opin. Ther. patents 14, 1403-1420 have been described.

In fact, this finding was confirmed by biochemical studies in which the effect of certain nonsteroidal anti-inflammatory drugs (NSAIDs) on gamma-secretase was demonstrated (US 2002/0128319; Eriksen (2003) J. Clin. invest.112, 440). Potential limitations of using NSAIDs for the prevention or treatment of AD are their Cyclooxygenase (COX) inhibitory activity, which can lead to unwanted side effects, and their low CNS penetration (Peretto et al, 2005, j.med.chem.48, 5705-. Recently, the NSAID R-flurbiprofen (flurbiprofen), an enantiomer lacking Cox-inhibiting activity and associated gastric toxicity, failed in a large third phase trial because the drug did not significantly improve the patient's ability to think and perform daily activities compared to patients receiving placebo.

WO-2009/032277 relates to heterocyclic compounds useful as modulators of gamma secretase.

US 2008/0280948 a1 relates to aminophenyl derivatives which are modulators of amyloid β.

WO-2009/005729 relates to heterocyclic compounds and their use as gamma secretase modulators.

There is a strong need for new compounds that can modulate gamma-secretase activity, thus opening new avenues for treating alzheimer's disease. It is an object of the present invention to overcome or ameliorate at least one of the disadvantages of the prior art, or to provide a useful alternative. It is therefore an object of the present invention to provide such novel compounds.

Brief description of the invention

The compounds of the present invention have been found to be useful as gamma secretase modulators. The compounds according to the invention, and pharmaceutically acceptable compositions thereof, may be used in the treatment or prevention of alzheimer's disease.

The present invention relates to novel compounds of formula (I):

and stereoisomeric forms thereof, wherein

R¹Is optionally substituted by one or more radicals each independently selected from halo, C_1-4Alkoxy radical, C_3-7C substituted by substituents of cycloalkyl, tetrahydropyranyl, tetrahydrofuranyl and phenyl_1-6An alkyl group; c_3-7Cycloalkyl, tetrahydropyranyl, tetrahydrofuranyl, 1, 3-benzodioxolyl or phenyl;

wherein each phenyl is independently optionally substituted with one or more substituents each independently selected from halo, cyano, C optionally substituted with one or more halo substituents_1-4Alkyl, andc optionally substituted by one or more halo substituents_1-4Substituent substitution of alkoxy;

R²is hydrogen, cyano or optionally substituted by one or more radicals each independently selected from C_1-4Alkoxy, halo and NR³R⁴C substituted by a substituent of_1-4An alkyl group;

X¹is CH or N;

X²is CR⁵Or N;

R⁵is hydrogen, halo, cyano, C_1-4Alkoxy or optionally substituted by one or more radicals each independently selected from halo, C_1-4Alkoxy and NR³R⁴C substituted by a substituent of_1-4An alkyl group;

X³is CR₆Or N;

R⁶is hydrogen, halo, cyano, C_1-4Alkoxy or optionally substituted by one or more radicals each independently selected from halo, C_1-4Alkoxy and NR³R⁴C substituted by a substituent of_1-4An alkyl group;

wherein each R³Independently of one another is hydrogen, C_1-4Alkyl or C_1-4An acyl group;

wherein each R⁴Independently of one another is hydrogen, C_1-4Alkyl or C_1-4An acyl group;

provided that X is¹，X²And X³No more than two of which are N;

A¹is CR⁷Or N; wherein R is⁷Is hydrogen, halo or C_1-4An alkoxy group;

A²，A³and A⁴Each independently is CH or N; with the proviso that A¹，A²，A³And A⁴No more than two of which are N;

Het¹article for making5-membered aromatic heterocycles having the formula (a-1), (a-2), (a-3) or (a-4)

R⁸Is hydrogen or C_1-4An alkyl group;

R⁹is hydrogen or C_1-4An alkyl group;

R¹⁰is hydrogen or C_1-4An alkyl group;

R¹¹is hydrogen or C_1-4An alkyl group;

R¹²is C_1-4An alkyl group;

G¹is O or S;

G²is CH or N;

and pharmaceutically acceptable addition salts and solvates thereof.

The invention also relates to processes for the preparation of compounds of formula I and pharmaceutical compositions containing them.

It has surprisingly been found that the compounds of the present invention modulate γ -secretase activity in vitro and in vivo and are therefore useful in the treatment or prevention of Alzheimer's Disease (AD), Traumatic Brain Injury (TBI), Mild Cognitive Impairment (MCI), senility, dementia associated with lewy bodies, amyloid cerebrovascular disease, multi-infarct dementia, down's syndrome, dementia associated with parkinson's disease and dementia associated with beta-amyloid, preferably AD and other diseases associated with beta-amyloid pathologies, such as glaucoma.

In view of the above-mentioned pharmacological properties of the compounds of formula (I), they have been shown to be suitable for use as medicaments.

More specifically, the compounds are suitable for the treatment or prophylaxis of Alzheimer's disease, amyloid cerebrovascular disease, multi-infarct dementia, dementia pugilistica or Down's syndrome.

The invention also relates to compounds according to general formula (I), the stereoisomeric forms and the pharmaceutically acceptable acid or base addition salts and the solvates thereof, for use in modulating the activity of gamma-secretase.

The invention will now be further illustrated. In the following paragraphs, the different aspects of the invention are defined in more detail. Aspects so defined may be combined with any other aspect or aspects unless clearly indicated to the contrary. In particular, any feature that is recited as being preferred or advantageous may be combined with any feature or features that are recited as being preferred or advantageous.

Detailed Description

When describing the compounds of the present invention, the terms used are to be construed in accordance with the following definitions, unless the context indicates otherwise.

When the term "substituted" is used in the present invention, unless otherwise indicated or clear from context, it is intended that one or more hydrogens, particularly from 1 to 4 hydrogens, preferably from 1 to 3 hydrogens, more preferably 1 hydrogen, on the atom or group to which the expression "substituted" is applied is replaced with a group selected from the group indicated, provided that the normal valency is not exceeded, and that the substitution results in a chemically stable compound, i.e., a compound that is sufficiently stable to survive isolation from the reaction mixture to a useful purity and formulation into a therapeutic agent.

The terms "halo" or "halogen" as a group or part of a group refer broadly to fluoro, chloro, bromo, iodo, unless otherwise indicated.

The term "C_1-6When alkyl "is a group or part of a group, it refers to formula C_nH_2n+1Wherein n is a number from 1 to 6. C_1-6The alkyl group contains 1 to 6 carbon atoms, preferably 1 to 4 carbon atoms, more preferably 1 to 3 carbon atoms, even more preferably 1 to 2 carbon atoms. The alkyl group may be straight or branched and may be substituted as described hereinAnd (4) substitution. When a subscript is used herein after a carbon atom, the subscript refers to the number of carbon atoms that the group referred to may contain. Thus, for example, C_1-6Alkyl includes all straight or branched chain alkyl groups containing between 1 and 6 carbon atoms and thus includes, for example, methyl, ethyl, n-propyl, isopropyl, 2-methyl-ethyl, butyl and its isomers (e.g., n-butyl, isobutyl and tert-butyl), pentyl and its isomers, hexyl and its isomers and the like.

The term "C_1-4When alkyl "is a group or part of a group, it refers to formula C_nH_2n+1Wherein n is a number from 1 to 4.C_1-4The alkyl group contains 1 to 4 carbon atoms, preferably 1 to 3 carbon atoms, more preferably 1 to 2 carbon atoms. Alkyl groups may be straight or branched chain and may be substituted as described herein. When a subscript is used herein after a carbon atom, the subscript refers to the number of carbon atoms that the group referred to may contain. Thus, for example, C_1-4Alkyl includes all straight or branched chain alkyl groups containing between 1 and 4 carbon atoms and thus includes, for example, methyl, ethyl, n-propyl, isopropyl, 2-methyl-ethyl, butyl and isomers thereof (e.g., n-butyl, isobutyl and tert-butyl), and the like.

The term "C_1-4Acyl "alone or in combination refers to a group containing 1 to 4 carbon atoms, wherein the carbonyl group is attached to hydrogen or a straight or branched chain hydrocarbon group containing 1 to 3 carbon atoms. Suitable C_1-4Non-limiting examples of acyl groups include formyl, acetyl, propionyl, butyryl and isobutyryl.

The term "C_1-4When an alkoxy group is a group OR part of a group, it is intended to have the formula-OR^cWherein R is^cIs C_1-4An alkyl group. Suitable C_1-4Non-limiting examples of alkoxy groups include methyloxy (i.e., methoxy), ethyloxy (i.e., ethoxy), propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, and tert-butoxy.

The term "C_3-7Cycloalkyl "alone or in combination, means containing 3Cyclic saturated hydrocarbon groups of up to 7 carbon atoms. Suitable C_3-7Non-limiting examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

The Chemical nomenclature of the compounds of the present invention is generated according to the nomenclature rules agreed upon by Chemical abstracts service.

In tautomeric forms, it should be clear that other tautomeric forms not described are also included within the scope of the invention.

When any variable occurs more than one time in any constituent, each definition is independent.

It will be appreciated that certain compounds of formula (I) and the pharmaceutically acceptable addition salts and stereoisomeric forms thereof may contain one or more chiral centers and exist as stereoisomeric forms.

The term "stereoisomeric forms" as used above defines all possible isomeric forms which the compounds of formula (I) have. Unless otherwise mentioned or indicated, the chemical designation of compounds represents a mixture of all possible stereochemically isomeric forms. More specifically, stereogenic centers may have either the R-or S-configuration; the substituents on the divalent cyclic (partially) saturated groups may have either the cis-or trans-configuration. Compounds containing a double bond may have E-or Z-stereochemistry at the double bond. Stereoisomeric forms of the compounds of formula (I) are included within the scope of the present invention.

When a particular stereoisomeric form is indicated, this means that said form is substantially free, i.e. associated with less than 50%, preferably less than 20%, more preferably less than 10%, even more preferably less than 5%, further preferably less than 2%, and most preferably less than 1% of other isomers.

For medical use, salts of the compounds of formula (I) are those wherein the counterion is pharmaceutically acceptable. However, salts of non-pharmaceutically acceptable acids and bases may also find use, for example, in the preparation or purification of pharmaceutically acceptable compounds. All salts, whether pharmaceutically acceptable or not, are included within the scope of the present invention.

The pharmaceutically acceptable acid or base addition salts mentioned above or below are meant to be the therapeutically active non-toxic acid and base addition salt forms which the compounds of formula (I) may form. Pharmaceutically acceptable acid addition salts are conveniently obtained by treating the base form with such a suitable acid. Suitable acids include, for example, inorganic acids such as hydrohalic acids, e.g., hydrochloric or hydrobromic acids, sulfuric acid, nitric acid, phosphoric acid, and the like; or organic acids such as acetic, propionic, glycolic, lactic, pyruvic, oxalic (i.e., oxalic), malonic, succinic (i.e., succinic), maleic, fumaric, malic, tartaric, citric, methanesulfonic, ethanesulfonic, benzenesulfonic, p-toluenesulfonic, cyclohexanoic, salicylic, p-aminosalicylic, pamoic and the like acids. Conversely, the salt form may be converted to the free base form by treatment with a suitable base.

The compounds of formula (I) containing an acidic proton may also be converted into their non-toxic metal or amine addition salt forms by treatment with suitable organic and inorganic bases. Suitable base salt forms include, for example, the ammonium salts, alkali metal and alkaline earth metal salts, such as the lithium, sodium, potassium, magnesium, calcium salts and the like, salts with organic bases, such as aliphatic and aromatic primary, secondary and tertiary amines, such as methylamine, ethylamine, propylamine, isopropylamine, the four butylamine isomers, dimethylamine, diethylamine, diethanolamine, dipropylamine, diisopropylamine, di-n-butylamine, pyrrolidine, piperidine, morpholine, trimethylamine, triethylamine, tripropylamine, quinuclidine, pyridine, quinoline and isoquinoline; benzathine, N-methyl-D-glucamine, hydrabamine salt, and salts with amino acids such as arginine, lysine, and the like. Conversely, the salt form may be converted to the free acid form by treatment with an acid.

The term solvate includes hydrates and solvent addition forms which the compounds of formula (I) are able to form, as well as salts thereof. Examples of such forms are e.g. hydrates, alcoholates and the like.

The compounds of formula (I) prepared in the processes described below can be synthesized in the form of racemic mixtures of enantiomers, which can be separated from each other according to resolution methods known in the art. A method of separating the enantiomeric forms of the compounds of formula (I) comprises liquid chromatography using a chiral stationary phase. The pure stereochemically isomeric forms may also be derived from the corresponding pure stereochemically isomeric forms of the appropriate starting materials, provided that the reaction occurs stereospecifically. Preferably, if a particular stereoisomer is desired, the compound will be synthesized by stereospecific methods of preparation. These methods would facilitate the use of enantiomerically pure starting materials.

In the framework of this application, the compounds according to the invention are all isotopic combinations inherently intended to include their chemical elements. In the framework of this application, a chemical element, in particular when mentioned in the compounds according to formula (I), includes all isotopes and isotopic mixtures of this element. For example, when reference is made to hydrogen, this is to be understood as meaning¹H，²H，³H and mixtures thereof.

Thus, compounds according to the present invention inherently include compounds containing one or more isotopes of one or more elements, and mixtures thereof, including radioactive compounds, also known as radiolabeled compounds, in which one or more non-radioactive atoms are replaced by one of its radioactive isotopes. The term "radiolabeled compound" means any compound according to formula (I) or a pharmaceutically acceptable salt thereof, which contains at least one radioactive atom. For example, the compounds may be labeled with a positron or gamma emitting radioisotope. For use in radioligand binding techniques,³h-atom or¹²⁵The I-atom is a selection of atoms that are substituted. For imaging, the most commonly used Positron Emitting (PET) radioisotope is¹¹C，¹⁸F，¹⁵O and¹³n, which are all accelerator production and have half-lives of 20, 100, 2 and 10 minutes, respectively. Because of the short half-life of these radioisotopes, it is only feasible that a mechanism with an accelerator can use them on siteIn their production, and therefore their use is limited. The most widely used of these isotopes is¹⁸F，^99mTc，²⁰¹Tl and¹²³I. the skilled worker is familiar with the handling of these radioisotopes, their production, isolation and incorporation into molecules.

Specifically, the radioactive atoms are selected from the group consisting of hydrogen, carbon, nitrogen, sulfur, oxygen, and halogens. In particular, the radioisotope is selected from³H，¹¹C，¹⁸F，¹²²I，¹²³I，¹²⁵I，¹³¹I，⁷⁵Br，⁷⁶Br，⁷⁷Br and⁸²Br。

as used in the specification and the appended claims, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. For example, "a compound" refers to one compound or more than one compound.

The terms described above and others used in the specification are well known to those skilled in the art.

Preferred features of the compounds of the invention are now set out.

The invention relates in particular to novel compounds of formula (I):

and stereoisomeric forms thereof, wherein

R¹Is optionally substituted by one or more radicals each independently selected from halo, C_1-4Alkoxy radical, C_3-7C substituted by substituents of cycloalkyl, tetrahydropyranyl, tetrahydrofuranyl and phenyl_1-6An alkyl group;

C_3-7cycloalkyl, tetrahydropyranyl, tetrahydrofuranyl, 1, 3-benzodioxolyl or phenyl;

wherein each phenyl is independently optionally substituted with one or more substituents each independently selected from halo, cyano, C optionally substituted with one or more halo substituents_1-4Alkyl, and C optionally substituted with one or more halo substituents_1-4Substituent substitution of alkoxy;

X¹is CH or N;

X²is CR⁵Or N;

X³is CR⁶Or N;

provided that X is¹，X²And X³No more than two of which are N;

A¹is CR⁷Or N; wherein R is⁷Is hydrogen, halo or C_1-4An alkoxy group;

Het¹is a 5-membered aromatic heterocycle having the formula (a-1), (a-2), (a-3) or (a-4)

R⁸Is hydrogen or C_1-4An alkyl group;

R⁹is hydrogen or C_1-4An alkyl group;

R¹⁰is hydrogen or C_1-4An alkyl group;

R¹¹is hydrogen or C_1-4An alkyl group;

R¹²is C_1-4An alkyl group;

G¹is O or S;

G²is CH or N;

and pharmaceutically acceptable addition salts and solvates thereof.

One embodiment of the present invention relates to compounds of formula (I)

And stereoisomeric forms thereof, wherein

C_3-7a cycloalkyl group, a tetrahydropyranyl group,tetrahydrofuranyl, 1, 3-benzodioxolyl or phenyl; wherein each phenyl is independently optionally substituted with one or more substituents each independently selected from halo, cyano, and C_1-4Alkyl, and C optionally substituted with one or more substituents each independently selected from halo_1-4Substituent substitution of alkoxy;

R²is hydrogen, cyano, optionally substituted by one or more radicals each independently selected from C_1-4Alkoxy, halo and NR³R⁴C substituted by a substituent of_1-4An alkyl group;

X¹is CH or N;

X²is CR⁵Or N;

X³is CR⁶Or N;

provided that X is¹，X²And X³No more than two of which are N;

A¹is CR⁷Or N; wherein R is⁷Is hydrogen, halo or C_1-4An alkoxy group;

R⁸Is hydrogen or C_1-4An alkyl group;

R⁹is hydrogen or C_1-4An alkyl group;

R¹⁰is hydrogen or C_1-4An alkyl group;

R¹¹is hydrogen or C_1-4An alkyl group;

R¹²is C_1-4An alkyl group;

G¹is O or S;

G²is CH or N;

and pharmaceutically acceptable addition salts and solvates thereof.

Another embodiment of the present invention relates to those compounds of formula (I) or any subgroup mentioned in any other embodiment, wherein one or more, preferably all, of the following restrictions apply:

(a)R¹is optionally substituted by one or more radicals each independently selected from halo, C_1-4Alkoxy radical, C_3-7Cycloalkyl and phenyl substituted C_1-6An alkyl group;

C_3-7cycloalkyl, tetrahydropyranyl, 1, 3-benzodioxolyl or phenyl;

wherein each phenyl group is independently substituted by one or moreEach independently selected from halo, C_1-4Alkyl and C_1-4Substituent substitution of alkoxy;

(b)R²is hydrogen, cyano or optionally substituted by one or more NH groups₂C substituted by substituents_1-4An alkyl group;

(c)X²is CR⁵Or N; in particular X²Is CR⁵；

(d)R⁵Is hydrogen, halo, cyano or optionally substituted by one or more NH groups₂C substituted by substituents_1-4An alkyl group;

(e)X³is CH or N;

(f)A²is CH or N and A³And A⁴Is CH; in particular A²，A³And A⁴Is CH;

(g)Het¹is a 5-membered aromatic heterocycle of the formula (a-1), (a-2), (a-3) or (a-4), in particular Het¹Is a 5-membered aromatic heterocycle having the formula (a-1), (a-2) or (a-3);

(h)R¹⁰is C_1-4An alkyl group;

(i)R¹¹is hydrogen;

(j)R⁸is hydrogen;

(k)R¹²is C_1-4An alkyl group.

(a)R¹is C optionally substituted with one or more substituents each independently selected from the group consisting of fluoro, methoxy, cyclopropyl and phenyl_1-4An alkyl group;

cyclobutyl, tetrahydropyranyl, 1, 3-benzodioxolyl or phenyl;

wherein each phenyl group is independently substituted by one or more groups each independently selected from methoxy, ethoxy, C_1-4Alkyl and fluoro;

(b)R²is hydrogen, cyano, optionally substituted by an NH group₂A substituent-substituted methyl group;

(c)X²is CR⁵Or N; in particular X²Is CR⁵；

(d)R⁵Is hydrogen, fluorine, cyano, optionally substituted by an NH group₂A substituent-substituted methyl group;

(e)X³is CH or N;

(f)R⁷is hydrogen, fluoro or methoxy;

(g)A²is CH or N and A³And A⁴Is CH; in particular A²，A³And A⁴Is CH;

(h)Het¹is a 5-membered aromatic heterocycle having the formula (a-1), (a-2), (a-3) or (a-4), in particular of the formula (a-1), (a-2) or (a-3);

(i)R¹⁰is methyl;

(j)R¹¹is hydrogen;

(k)R⁸is hydrogen;

(l)R¹²is methyl.

(a)R¹is covered by a C_1-4Phenyl substituted with an alkoxy substituent; or R¹Is C substituted by one or more halo substituents_1-6An alkyl group;

(b)R²is hydrogen;

(c)X¹，X²and X³Is CH;

(d)A¹is CR⁷(ii) a Wherein R is⁷Is C_1-4An alkoxy group;

(e)A²，A³and A⁴Is CH;

(f)Het¹has the formula (a-1) or (a-2);

(g)G¹is O;

(h)G²is CH;

(i)R⁸is C_1-4An alkyl group;

(j)R¹⁰is C_1-4An alkyl group;

(k)R⁹is hydrogen.

(a)R¹is a quilt C_1-4Phenyl substituted with an alkoxy substituent;

(b)R²is hydrogen;

(c)X¹，X²and X³Is CH;

(d)A¹is CR⁷(ii) a Wherein R is⁷Is C_1-4An alkoxy group;

(e)A²，A³and A⁴Is CH;

(f)Het¹having formula (a-2);

(g)G²is CH;

(h)R¹⁰is C_1-4An alkyl group;

(i)R⁹is hydrogen.

(a)R¹is C substituted by one or more halo substituents_1-6An alkyl group; in particular R¹Is C substituted by 3 halo substituents_1-6An alkyl group;

(b)R²is hydrogen;

(c)X¹，X²and X³Is CH;

(d)A¹is CR⁷(ii) a Wherein R is⁷Is C_1-4An alkoxy group;

(e)A²，A³and A⁴Is CH;

(f)Het¹having formula (a-1);

(g)G¹is O;

(h)R⁸is C_1-4An alkyl group;

(i)R⁹is hydrogen.

In a further embodiment, the invention relates to a compound according to any other embodiment, wherein R is¹Is C optionally substituted by one or more substituents each independently selected from optionally substituted by one or more halo substituents_1-4Alkyl and C optionally substituted by one or more halo substituents_1-4Phenyl substituted with a substituent of alkoxy.

In a further embodiment, the invention relates to a compound according to any other embodiment, wherein R is¹Is selected from one or more of C independently_1-4Alkyl and C_1-4Phenyl substituted with a substituent of alkoxy.

In a further embodiment, the invention relates to a method according to any of the other embodimentsCompound (I) wherein R¹Is C optionally substituted by one or more halo substituents_1-6An alkyl group.

In a further embodiment, the invention relates to a compound according to any other embodiment, wherein R is¹Is C optionally substituted by one or more fluoro substituents_1-6An alkyl group.

In a further embodiment, the invention relates to a compound according to any other embodiment, wherein R is¹Is 2, 2, 2-trifluoroethyl.

In a further embodiment, the invention relates to a compound according to any other embodiment, wherein X is¹Is CH.

In a further embodiment, the invention relates to a compound according to any other embodiment, wherein X is¹Is N.

In a further embodiment, the invention relates to a compound according to any other embodiment, wherein X is²Is CH.

In a further embodiment, the invention relates to a compound according to any other embodiment, wherein X is²Is N.

In a further embodiment, the invention relates to a compound according to any other embodiment, wherein X is³Is N.

In a further embodiment, the invention relates to a compound according to any other embodiment, wherein X is³Is CR⁶。

In a further embodiment, the invention relates to a compound according to any other embodiment, wherein R is⁶Is hydrogen.

In a further embodiment, the invention relates to a compound according to any other embodiment, wherein a¹Is CR⁷。

In further onIn an embodiment, the invention relates to compounds according to any other embodiment, wherein a¹Is N.

In a further embodiment, the invention relates to a compound according to any other embodiment, wherein a²，A³And A⁴Each independently is CH.

In a further embodiment, the invention relates to a compound according to any other embodiment, wherein Het is¹Has the formula (a-3).

In a further embodiment, the invention relates to a compound according to any other embodiment, wherein Het is¹Has the formula (a-4).

In a further embodiment, the invention relates to a compound according to any other embodiment, wherein G is¹Is S.

In a further embodiment, the invention relates to a compound according to any other embodiment, wherein G is²Is N.

Another embodiment of the invention relates to those compounds of formula (I) or any subgroup mentioned in any other embodiment, wherein the 1, 3-benzodioxolyl group is limited to the 1, 3-benzodioxol-5-yl group.

In one embodiment, the compound of formula (I) is selected from:

n- [ 3-methoxy-4- (4-methyl-1H-imidazol-1-yl) phenyl ] -2- (3-methoxyphenyl) -2H-indazol-7-amine,

2- [ (4-fluorophenyl) methyl ] -N- [ 3-methoxy-4- (4-methyl-1H-imidazol-1-yl) phenyl ] -2H-indazol-7-amine,

n- [ 3-methoxy-4- (4-methyl-1H-imidazol-1-yl) phenyl ] -2- (3-methoxyphenyl) -3-methyl-2H-indazol-7-amine,

n- [ 3-methoxy-4- (3-methyl-1H-1, 2, 4-triazol-1-yl) phenyl ] -2- (3-methoxyphenyl) -2H-indazol-7-amine,

2-butyl-N- [ 3-methoxy-4- (4-methyl-1H-imidazol-1-yl) phenyl ] -2H-indazol-7-amine,

2-butyl-N- [ 3-methoxy-4- (4-methyl-1H-imidazol-1-yl) phenyl ] -2H-indazol-7-amine, 2HCl,

2-butyl-N- [ 3-methoxy-4- (4-methyl-1H-imidazol-1-yl) phenyl ] -3-methyl-2H-indazol-7-amine,

2-butyl-N- [ 3-methoxy-4- (4-methyl-1H-imidazol-1-yl) phenyl ] -3-methyl-2H-indazol-7-amine, 2HCl,

2- (4-fluorophenyl) -N- [ 3-methoxy-4- (2-methyl-5-)Azolyl) phenyl]-3-methyl-2H-indazol-7-amine,

2- (3-methoxyphenyl) -3-methyl-N- [6- (4-methyl-5-)Azolyl) -3-pyridinyl]-2H-indazol-7-amine,

2- (4-fluorophenyl) -3-methyl-N- [6- (4-methyl-5-)Azolyl) -3-pyridinyl]-2H-indazol-7-amine,

n- [ 3-fluoro-4- (3-methyl-1H-1, 2, 4-triazol-1-yl) phenyl ] -3-methyl-2- (4, 4, 4-trifluorobutyl) -2H-indazol-7-amine,

n- [ 3-methoxy-4- (4-methyl-1H-imidazol-1-yl) phenyl ] -2-methyl-2H-indazol-7-amine,

2- (4-fluorophenyl) -3-methyl-N- [4- (2-methyl-5-)Azolyl) phenyl]-2H-indazol-7-amine,

2- (4-fluorophenyl) -3-methyl-N- [4- (2-methyl-5-)Azolyl) phenyl]-2H-indazol-7-amine 1.5HCl,

2- (3-methoxyphenyl) -3-methyl-N- [4- (2-methyl-5-)Azolyl) phenyl]-2H-indazol-7-amine,

2- (2, 4-difluorophenyl) -N- [ 3-methoxy-4- (4-methyl-1H-imidazol-1-yl) phenyl ] -3-methyl-2H-indazol-7-amine,

2- (2, 4-difluorophenyl) -3-methyl-N- [4- (2-methyl-5-)Azolyl) phenyl]-2H-indazol-7-amine,

2- [ 4-ethoxy-2-methyl-5- (1-methylethyl) phenyl]-3-methyl-N- [4- (2-methyl-5-)Azolyl) phenyl]-2H-indazol-7-amine,

2- (2, 4-difluorophenyl) -N- [ 3-fluoro-4- (3-methyl-1H-1, 2, 4-triazol-1-yl) phenyl ] -3-methyl-2H-indazol-7-amine,

2- [ 4-ethoxy-2-methyl-5- (1-methylethyl) phenyl ] -N- [ 3-methoxy-4- (4-methyl-1H-imidazol-1-yl) phenyl ] -3-methyl-2H-indazol-7-amine,

2- [ 4-ethoxy-2-methyl-5- (1-methylethyl) phenyl ] -N- [ 3-methoxy-4- (3-methyl-1H-1, 2, 4-triazol-1-yl) phenyl ] -3-methyl-2H-indazol-7-amine,

2- (2, 4-difluorophenyl) -N- [ 3-methoxy-4- (2-methyl-5-thiazolyl) phenyl ] -3-methyl-2H-indazol-7-amine,

n- [ 3-methoxy-4- (4-methyl-5-)Azolyl) phenyl]-2- (3-methoxyphenyl) -3-methyl-2H-indazol-7-amine,

n- [ 3-methoxy-4- (4-methyl-5-)Azolyl) phenyl]-2- (3-methoxyphenyl) -3-methyl-2H-indazol-7-amine 1.9HCl,

n- [ 3-methoxy-4- (3-methyl-1H-1, 2, 4-triazol-1-yl) phenyl ] -2- (3-methoxyphenyl) -3-methyl-2H-indazol-7-amine,

n- [ 3-methoxy-4- (3-methyl-1H-1, 2, 4-triazol-1-yl) phenyl ] -2- (3-methoxyphenyl) -3-methyl-2H-indazol-7-amine 1.9HCl,

n- [ 3-methoxy-4- (3-methyl-1H-1, 2, 4-triazol-1-yl) phenyl ] -2-methyl-2H-indazol-7-amine,

2-methyl-N- [4- (2-methyl-5-)Azolyl) phenyl]-2H-indazol-7-amine,

n- [ 3-methoxy-4- (2-methyl-5-)Azolyl) phenyl]-2- (3-methoxyphenyl) -3-methyl-2H-pyrazolo [3, 4-c](ii) a pyridine-7-amine,

2- (cyclopropylmethyl) -N- [ 3-methoxy-4- (4-methyl-1H-imidazol-1-yl) phenyl ] -2H-indazol-7-amine,

2- (cyclopropylmethyl) -N- [ 3-methoxy-4- (4-methyl-1H-imidazol-1-yl) phenyl ] -2H-indazol-7-amine, 2HCl,

n- [ 3-methoxy-4- (2-methyl-5-)Azolyl) phenyl]-2- (2, 2, 2-trifluoroethyl) -2H-indazol-7-amine,

n- [4- [2- (1-methylethyl) -5-Azolyl radical]Phenyl radical]-2- (2, 2, 2-trifluoroethyl) -2H-indazol-7-amine,

n- [ 3-methoxy-4- (2-methyl-5-thiazolyl) phenyl ] -2-methyl-2H-indazol-7-amine,

2-butyl-7- [ [ 3-methoxy-4- (4-methyl-1H-imidazol-1-yl) phenyl ] amino ] -2H-indazole-5-carbonitrile,

2-butyl-7- [ [ 3-methoxy-4- (4-methyl-1H-imidazol-1-yl) phenyl ] amino ] -2H-indazole-5-carbonitrile.2 HCl,

2-cyclobutyl-N- [4- (2-methyl-5-)Azolyl) phenyl]-2H-indazol-7-amine,

2-cyclobutyl-N- [4- (2-methyl-5-)Azolyl) phenyl]-2H-indazol-7-amine 1.2HCl,

2- (4-fluorophenyl) -7- [ [4- (2-methyl-5-)Azolyl) phenyl]Amino group]-2H-indazole-3-carbonitrile,

2- (2-methoxyethyl) -N- [4- (2-methyl-5-)Azolyl) phenyl]-2H-indazol-7-amine,

2- (2-methoxyethyl) -N- [4- (2-methyl-5-)Azolyl) phenyl]-2H-indazol-7-amine 1.5HCl.1.25H₂O，

2- (2-methoxyethyl) -N- [ 3-methoxy-4- (4-methyl-5-)Azolyl) phenyl]-2H-indazol-7-amine,

2- (2-methoxyethyl) -N- [ 3-methoxy-4- (4-methyl-5-)Azolyl) phenyl]-2H-indazol-7-amine 1.5HCl.0.18H₂O，

2- (cyclopropylmethyl) -N- [4- (2-methyl-5-)Azolyl) phenyl]-2H-indazol-7-amine,

2- (cyclopropylmethyl) -N- [4- (2-methyl-5-)Azolyl) phenyl]-2H-indazol-7-amine 2HCl,

2- (1, 3-benzodioxol-5-yl) -N- [4- (2-methyl-5-)Azolyl) phenyl]-2H-indazol-7-amine,

2- (cyclopropylmethyl) -N- [ 3-methoxy-4- (4-methyl-5-)Azolyl) phenyl]-2H-indazol-7-amine,

2- (cyclopropylmethyl) -N- [ 3-methoxy-4- (4-methyl-5-)Azolyl) phenyl]-2H-indazol-7-amine 2HCl,

2- (3-methoxyphenyl) -3-methyl-N- [4- (2-methyl-5-)Azolyl) phenyl]-2H-pyrazolo [3, 4-c]Pyridine compound-a (a) 7-amine,

n- [ 3-fluoro-4- (1-methyl-1H-pyrazol-4-yl) phenyl ] -2- (3-methoxyphenyl) -3-methyl-2H-indazol-7-amine,

2- (cyclopropylmethyl) -N- [ 3-methoxy-4- (2-methyl-5-)Azolyl) phenyl]-2H-indazol-7-amine,

2- (cyclopropylmethyl) -N- [ 3-methoxy-4- (2-methyl-5-)Azolyl) phenyl]-2H-indazol-7-amine HCl,

2- (4-fluorophenyl) -7- [ [4- (2-methyl-5-)Azolyl) phenyl]Amino group]-2H-indazole-3-methanamine,

2-butyl-7- [ [ 3-methoxy-4- (4-methyl-1H-imidazol-1-yl) phenyl ] amino ] -2H-indazole-5-methanamine,

2-butyl-7- [ [ 3-methoxy-4- (4-methyl-1H-imidazol-1-yl) phenyl ] amino ] -2H-indazole-5-methanamine, 4HCl,

2- (cyclopropylmethyl) -N- [6- (2-methyl-5-)Azolyl) -3-pyridinyl 1]-2H-indazol-7-amine,

2- (cyclopropylmethyl) -N- [6- (2-methyl-5-)Azolyl) -3-pyridinyl 1]-2H-indazol-7-amine 2HCl,

n- [4- (2-methyl-5-)Azolyl) phenyl]-2- (tetrahydro-2H-pyran-4-yl) -2H-indazole-a (a) 7-amine,

n- [6- (2-methyl-5-)Azolyl) -3-pyridinyl]-2- (2, 2, 2-trifluoroethyl) -2H-indazol-7-amine,

n- [ 3-methoxy-4- (3-methyl-1H-1, 2, 4-triazol-1-yl) phenyl ] -2- (2, 2, 2-trifluoroethyl) -2H-indazol-7-amine,

n- [ 3-methoxy-4- (2-methyl-5-)Azolyl) phenyl]-5-methyl-2- (2, 2, 2-trifluoroethyl) -2H-pyrazolo [3, 4-c](ii) a pyridine-7-amine,

n- [ 3-methoxy-4- (3-methyl-1H-1, 2, 4-triazol-1-yl) phenyl ] -5-methyl-2- (2, 2, 2-trifluoroethyl) -2H-pyrazolo [3, 4-c ] pyridin-7-amine,

2- (5-methoxy-2-methylphenyl) -3-methyl-N- [6- (2-methyl-5-)Azolyl) -3-pyridinyl]-2H-indazol-7-amine,

5-fluoro-2- (4-fluorophenyl) -3-methyl-N- [6- (2-methyl-5-)Azolyl) -3-pyridinyl]-2H-indazol-7-amine,

n- [ 3-methoxy-4- (2-methyl-5-)Azolyl) phenyl]-5-methyl-2- (2, 2, 2-trifluoroethyl) -2H-pyrazolo [4, 3-b](ii) a pyridine-7-amine,

n- [ 3-fluoro-4- (4-methyl-1H-imidazol-1-yl) phenyl ] -2- (2, 2, 2-trifluoroethyl) -2H-indazol-7-amine,

2- (3-methoxyphenyl) -3-methyl-N- [4- (1-methyl-1H-pyrazol-5-yl) phenyl ] -2H-indazol-7-amine,

2- (3-methoxyphenyl) -3-methyl-N- [4- (1-methyl-1H-pyrazol-5-yl) phenyl]-2H-indazol-7-amine 2HCl.0.5H₂O，

N- [ 6-methoxy-5- (4-methyl-1H-imidazol-1-yl) -2-pyridyl ] -2- (2, 2, 2-trifluoroethyl) -2H-indazol-7-amine,

n- [ 3-fluoro-4- (4-methyl-1H-imidazol-1-yl) phenyl ] -2- (2, 2, 2-trifluoroethyl) -2H-pyrazolo [3, 4-c ] pyridin-7-amine,

n- [ 3-methoxy-4- (3-methyl-1H-1, 2, 4-triazol-1-yl) phenyl ] -5- (1-methylethyl) -2- (2, 2, 2-trifluoroethyl) -2H-pyrazolo [4, 3-b ] pyridin-7-amine,

n- [ 6-methoxy-5- (4-methyl-1H-imidazol-1-yl) -2-pyridyl ] -5- (1-methylethyl) -2- (2, 2, 2-trifluoroethyl) -2H-pyrazolo [4, 3-b ] pyridin-7-amine,

including any stereochemically isomeric form thereof,

and pharmaceutically acceptable addition salts and solvates thereof.

In one embodiment, the compound of formula (I) is selected from: n- [ 3-methoxy-4- (2-methyl-5-)Azolyl) phenyl]-2- (2, 2, 2-trifluoroethyl) -2H-indazol-7-amine, and N- [ 3-methoxy-4- (4-methyl-1H-imidazol-1-yl) phenyl]-2- (3-methoxyphenyl) -3-methyl-2H-indazol-7-amine,

including any stereochemically isomeric form thereof,

and pharmaceutically acceptable addition salts and solvates thereof.

All possible combinations of the above-mentioned valuable embodiments are to be considered as being comprised within the scope of the present invention.

The invention also includes processes for the preparation of compounds of formula (I) and subgroups thereof. In the reactions described, it may be necessary to protect reactive functional groups, such as hydroxyl, amino or carboxyl groups, which are required in the final product, in order to avoid their unwanted participation in the reaction. Conventional protecting Groups may be used according to standard practice, see for example "protecting Groups in Organic Chemistry" by t.w.greene and p.g.m.wuts, John Wileyand Sons, 1999.

The compounds of formula (I) and subgroups thereof may be prepared by a series of steps as described hereinafter. They are generally prepared from starting materials which are commercially available or prepared by standard methods well known to those skilled in the art. The compounds of the invention can also be prepared using standard synthetic methods commonly used by those skilled in organic chemistry.

The general preparation of some typical examples is shown below. All variables are as defined above unless otherwise indicated. L is defined as a leaving group, such as Cl, Br, I, tosylate, mesylate or triflate, in particular Cl, Br or I, unless otherwise indicated.

Experimental method 1

In general, compounds of formula (I) may be prepared according to the statements of scheme 1 below, wherein all variables are as defined above:

the compounds of formula (I) may be prepared by coupling reactions between intermediates of formula (II) and (III), as shown in scheme 1, wherein all variables are as defined above. The reaction may be carried out in the presence of a suitable base such as Cs₂CO₃Or in the presence of sodium tert-butoxide. The reaction can be carried out in a reaction-inert solvent such as toluene, N, N-Dimethylformamide (DMF), tert-butanol or di-butanolIn an alkane. This is achieved byThe reaction is typically carried out in the presence of a catalyst system comprising a suitable catalyst such as palladium (II) acetate (Pd (OAc)₂) Or tris (dibenzylideneacetone) dipalladium (Pd)₂(dba)₃) And ligands such as (9, 9-dimethyl-9H-xanthene-4, 5-diyl) bis [ diphenylphosphine](Xantphos), [1, 1' -binaphthyl]-2, 2' -diylbis [ diphenylphosphine](BINAP) or dicyclohexyl [2 ', 4', 6 '-tris (1-methylethyl) [1, 1' -biphenyl ] group]-2-yl]-phosphines (X-phos). Preferably, the reaction is carried out under an inert atmosphere such as N₂Or under an Ar atmosphere. The reaction rate and yield can be increased by microwave-assisted heating.

Experimental method 2

Wherein R is²Is limited to C_1-4The alkyl intermediate of formula (III), hereinafter referred to as intermediate of formula (IV), may be prepared by alkylation of the intermediate of formula (V) according to conventional reaction methods generally known in the art. The alkylation reaction is carried out in the presence of a suitable base such as lithium diisopropylamide or lithium bis (trimethylsilyl) amide and an alkylating agent such as C_1-4In the presence of alkyl-Y (where Y is a reactive group such as Cl, Br or I). All other variables are as defined above. This reaction can be carried out in an aprotic solvent such as DMF or Tetrahydrofuran (THF).

Experimental method 3

Wherein R is²Restricted to-CH₂NH₂The intermediates of formula (III), hereinafter referred to as intermediates of formula (VI), may be prepared by reduction of intermediates of formula (VII) according to conventional reaction methods generally known in the art. This reaction is carried out in the presence of a suitable reducing agent, for example Raney Nickel. This reaction can be carried out in a protic solvent such as methanol (MeOH) in the presence of ammonia.

NH in intermediates of formula (VI)₂The radicals may be further alkylated and/or acylated to give further intermediates of the formula (III).

Experimental method 4

The intermediate of formula (V) may be prepared by reduction of the intermediate of formula (VIII) according to conventional reaction methods generally known in the art. The reduction may be carried out in a suitable reducing agent such as SnCl₂.H₂In the presence of O. This reaction can be carried out in a protic solvent, for example in ethanol (EtOH), at elevated temperatures, generally between 40 and 50 ℃.

In which X is¹Is defined as N and X²And X³In the particular case of intermediates of formula (V) defined as CH, hereinafter referred to as intermediates of formula (V-a), the 'L' group can be replaced by-OH or ethoxy groups by hydrolysis or ethanolysis. The intermediate thus obtained can be converted back again into the intermediate of formula (V-a) according to conventional reaction methods generally known in the art.

Experimental method 5

Intermediates of formula (VIII) can be prepared according to scheme 5 by reductive amination of intermediates of formula (IX). The reaction is carried out in a suitable reducing agent such as sodium triacetoxyborohydride (NaBH (OAc)₃) And primary amines such as R¹-NH₂In the presence of (a). This reaction can be carried out in an aprotic solvent such as 1, 2-dichloroethane.

Experimental method 6

As shown in scheme 6, intermediates of formula (IX) can be prepared by oxidation of intermediates of formula (X). The reaction is carried out in a suitable oxidizing agent such as sodium periodate (NaIO)₄) In the presence of oxygen. This reaction can be carried out in a mixture of solvents, for example water/DMF or water/THF.

Experimental method 7

The intermediate of formula (X) can be prepared by condensation of dimethylformamide dimethyl acetal (DMF-DMA) with the intermediate of formula (XI) as shown in scheme 7. Intermediate (XI) may be obtained commercially or may be prepared according to conventional reaction methods generally known in the art. Stirring and/or elevated temperature (e.g., between 70-110 ℃) can increase the rate of reaction.

Experimental method 8

Alternatively, intermediates of formula (IX) can also be prepared by oxidation of intermediates of formula (XII) according to scheme 8, which intermediates of formula (XII) are commercially available or can be prepared according to conventional reaction methods generally known in the art. The reaction is carried out in a suitable oxidizing agent such as manganese dioxide (MnO)₂) Or pyridinium chlorochromate(PCC) is present. The reaction may be carried out in a solvent such as Dichloromethane (DCM) or chloroform (CHCl)₃) Usually in the presence of molecular sieves.

Experimental method 9

Alternatively, intermediates of formula (V) may be prepared by alkylation of intermediates of formula (XIII) according to conventional reaction methods generally known in the art. The alkylation may be carried out in the absence or presence of a suitable base such as cesium carbonate or a primary amine such as N, N-dicyclohexyl-N-methylamine and an alkylating agent such as R¹-Y (wherein Y is defined as Cl, Br or I), R¹-O-SO₂R (wherein R may be selected from a wide variety of groups well known to those skilled in the art; a typical but non-limiting example of R is C_1-6Alkyl, perfluoro C_1-6Alkyl or optionally substituted phenyl; more particular examples of R are methyl or p-methylphenyl) or R¹-O-SO₂-R¹In the presence of oxygen. These alkylating agents are commercially available or may be prepared according to conventional reaction methods generally known in the art. This reaction can be carried out in a reaction-inert solvent such as toluene or DMF. Stirring, elevated temperature (e.g., between 70-110 ℃) can increase the reaction rate.

Experimental method 10

Intermediates of formula (XIII) can be prepared by deprotonation of intermediates of formula (XIV) according to conventional reaction methods generally known in the art. This reaction is usually carried out in a solvent such as Dimethylsulfoxide (DMSO) in the presence of a suitable base such as potassium tert-butoxide (KOtBu).

Experimental method 11

In the formula (XIV)The intermediates may be prepared by diazotization of the intermediates of formula (XV) according to conventional reaction methods generally known in the art. The reaction can be carried out in an aqueous acid solution, such as hydrochloric acid solution, in sodium nitrite (NaNO)₂) In the presence of oxygen. The reaction is usually carried out at low temperatures (< 5 ℃). The diazonium species is then quenched with tert-butyl mercaptan (t-BuSH) in a protic solvent such as EtOH at low temperatures (< 5 ℃).

Experimental method 12

Alternatively, intermediates of formula (XIII) may be prepared in a single step by diazotizing intermediates of formula (XV) according to conventional reaction methods generally known in the art. This reaction is usually carried out in an acidic solution, such as glacial acetic acid, in sodium nitrite (NaNO)₂) In the presence of an aqueous solution of (a).

Synthesis of an intermediate of formula (XIII), wherein X²Representing N, herein referred to as an intermediate of formula (XIII-a), requires a preliminary protection of the amino function of the intermediate of formula (XV), wherein X²Is N, as described in WO 2005/016892.

Experimental method 13

The intermediate of formula (VII) may be prepared by reduction of the intermediate of formula (XVI) according to conventional reaction methods generally known in the art. This reduction is usually carried out in a suitable reducing agent such as phosphorus trichloride (PCl)₃) Or triphenylphosphine. The reaction may be carried out in a reaction inert solvent such as CHCl₃At elevated temperatures (between 50 and 75 ℃).

Experimental method 14

The intermediates of formula (XVI) can be prepared by reacting an intermediate of formula (IX) with a primary amine R according to conventional reaction methods generally known in the art¹-NH₂Formed Schiff base. Treatment with sodium cyanide or Trimethylsilyl cyanide converts the schiff base into its α -aminonitrile derivative, which is then subjected to basic cyclization. The cyclisation step is carried out in the presence of a suitable base, such as an aqueous solution of sodium carbonate.

Experimental method 15

As shown in scheme 15, intermediates of formula (II) can be prepared by reduction of intermediates of formula (XVII), wherein all variables are as defined above.

The reduction of (XVII) to (II) may be carried out by conventional methods, for example reductive hydrogenation or reduction using a metal or metal salt and an acid [ e.g. a metal such as iron or metal salt such as SnCl₂And acids such as inorganic acids (hydrochloric acid, sulfuric acid, etc.) or organic acids (acetic acid, etc.)]Or other well-known methods for converting nitro groups to the corresponding amines.

Experimental method 16

Intermediates of formula (XVII) according to scheme 16 wherein Het¹Limited to (a-2) shown in scheme 16, herein referred to as intermediates of formula (XVIII), can be prepared by nucleophilic aromatic substitution reaction of intermediate (XIX) with optionally substituted imidazole or triazole of formula (XX), whichMiddle L^aIs defined as F, Cl or Br and wherein all other variables are as defined above. The reaction may be carried out under a protective atmosphere such as N₂The reaction is carried out under an atmosphere. Agitation, elevated temperature (e.g., between 70-170 ℃) and/or increased pressure may increase the reaction rate. The reaction can be carried out in an organic solvent such as DMSO, DMF or N-methylpyrrolidone (NMP) in a base such as K₂CO₃，Cs₂CO₃Or Et₃In the presence of N.

Intermediates of formula (XIX) and formula (XX) are commercially available or can be readily prepared by one skilled in the art.

Experimental method 17

Intermediates of formula (XVII) wherein Het¹Is limited to the 2-position via R as shown in scheme 17^8a(C_1-4Alkyl) substitutedOxazoles, referred to herein as intermediates of formula (XXI), can be prepared by condensation of intermediate of formula (XXII) (XIX) with an intermediate of formula (XXIII), which can be activated with iodobenzene diacetate in the presence of triflic acid. Stirring and/or increasing the temperature (e.g., between 70-100 ℃) can increase the reaction rate. In scheme 17, R^8aIs defined as C_1-4Alkyl and all other variables are as defined above.

Experimental method 18

Intermediates of formula (XXVII), according to scheme 18, wherein Het¹Is limited to the 4-position via R⁹Substituted byOxazoles, hereinafter referred to as intermediates of formula (XXIV), can be prepared by condensation of intermediates of formula (XXV) (XIX) with intermediates of formula (XXVI). Intermediate (XXVI) may be obtained commercially or may be prepared according to conventional reaction methods generally known in the art. This condensation reaction can generally be carried out in a suitable base such as K₂CO₃Or in the presence of sodium ethoxide (NaOEt). This reaction can be carried out in a protic solvent such as MeOH or EtOH. Agitation and/or increased temperature (e.g., between 70-110 c) can increase the reaction rate. In flow 18, all other variables are as defined above.

Alternatively, the reaction depicted in scheme 18 can also be carried out with benzaldehyde derivatives of intermediates of formula (XXVI), wherein NO is₂By Cl, Br or I.

Experimental method 19

According to scheme 19, intermediates of formula (II) can also be prepared according to well known reaction methods by converting the L-substituent of intermediates of formula (XXVII) to an amino group or a masked or protected amino function, which can then be converted to an amino group. In scheme 19, L^xDefined as Cl, Br or I and all other variables are as defined above.

Experimental method 20

According to scheme 20, compounds of formula (XVII) can also be prepared by coupling reactions between intermediates of formula (XVIII) and intermediates of formula (XXIX), wherein L^yDefined as Cl, Br or I and all other variables are as defined above.

In scheme 20, intermediates of formula (XXIX) can be obtained commercially or can be prepared according to conventional reaction methods generally known in the art. The coupling reaction may generally be carried out in the presence of a suitable base such as Cs₂CO₃，Na₂CO₃Or in the presence of CsF. The reaction may be carried out in a reaction-inert solvent such as toluene, DMF or bisIn an alkane. The reaction can be carried out in the presence of a catalyst such as tetrakis (triphenylphosphine) palladium (Pd (PPh)₃)₄) Or 1, 1-bis (diphenylphosphino ferrocene dichloropalladium II) (Pd (dppf) Cl₂) In the presence of oxygen. Agitation, elevated temperature (e.g., between 70-140 ℃) and/or increased pressure may increase the reaction rate. This reaction is preferably carried out under an inert atmosphere such as nitrogen or argon.

Alternatively, the boronic acid pinacol ester derivative of formula (XXIX) may be replaced with the corresponding boronic acid derivative.

Experimental method 21

Intermediates of formula (XVII) according to scheme 21 wherein Het¹Limited to that shown in scheme 21, hereinafter referred to as intermediates of formula (XXX), can be prepared by coupling reactions between intermediates of formula (XXXI) and intermediates of formula (XXXII), wherein L^bIs defined as I or Br and wherein all other variables are as defined above. In scheme 21, intermediates of formula (XXXI) and (XXXII) can be obtained commercially or can be prepared according to conventional reaction methods generally known in the art. The coupling reaction may be carried out in the presence of a suitable base such as Cs₂CO₃Or Ag₂CO₃In the presence of oxygen. The reaction may be carried out in a reaction-inert solvent such as H₂O，CH₃In CN or DMF. The reaction is usually carried out in the presence of a suitable catalyst, for example palladium (II) acetate (Pd (OAc)₂) Or 1 of the number of the groups in the group,1-bis (diphenylphosphino ferrocene dichloropalladium II) (Pd (dppf)) Cl₂) In the presence of a catalyst system and a ligand such as triphenylphosphine. Stirring, and increasing the temperature (e.g., between 60 and 140 ℃) can increase the reaction rate.

Experimental method 22

An intermediate of formula (XXVII) according to scheme 22, wherein Het¹Constrained to the scheme 22, hereinafter referred to as intermediates of formula (XXXIII), can be prepared by decarboxylation of compounds of formula (XXXIV), wherein L^xIs defined as Br, I or Cl and wherein all other variables are as defined above. The reaction can be carried out in a solvent such as quinoline or DMF, in the presence of copper (II) oxide (CuO), or in a DMF/EtOH mixture or isopropanol, both without CuO. The reaction can be carried out under microwave-assisted conditions. This reaction usually requires high temperatures (up to 150 ℃).

Experimental method 23

According to scheme 23, intermediates of formula (XXXIV) can be prepared by hydrolysis of a carboxylate functional group of a compound of formula (XXXV), wherein L^xIs defined as Br, I or Cl and wherein all other variables are as defined above. This reaction can be carried out under acidic conditions or basic conditions. Preferably under alkaline conditions in the presence of a base such as NaOH or LiOH at room temperatureIn a mixture of alkane and water.

Experimental method 24

According to scheme 24, intermediates of formula (XXXV) can be prepared by coupling reactions between intermediates of formula (XXXVI) and intermediates of formula (XXXVII), wherein L^xIs defined as Br, I or Cl, wherein L^cIs defined as Br or I, and wherein all other variables are as defined above. Intermediates of formula (XXXVI) and (XXXVII) can be obtained commercially or can be prepared according to conventional reaction methods generally known in the art. The coupling reaction is carried out in the presence of a suitable base such as Cs₂CO₃Or Ag₂CO₃In the presence of oxygen. The reaction may be carried out in a reaction-inert solvent such as CH₃CN, toluene or DMF. The reaction is usually carried out in the presence of a suitable catalyst, for example palladium (II) acetate (Pd (OAc)₂) Or [1, 1-bis (diphenylphosphino ferrocene) dichloropalladium II) (Pd (dppf) Cl₂) And a ligand such as triphenylphosphine or tri-o-tolylphosphine. Stirring, and increasing the temperature (e.g., between 60 and 140 ℃) can increase the reaction rate.

Experimental method 25

Wherein X²Restricted to CR⁵And R is⁵is-CH₂NH₂The intermediates of formula (III), hereinafter referred to as intermediates of formula (XXXVIII), may be prepared by reduction of intermediates of formula (XXXIX) according to conventional reaction methods generally known in the art. This reaction is carried out in the presence of a suitable reducing agent, for example Raney Nickel. This reaction can be carried out in a protic solvent, for example methanol (MeOH) in the presence of ammonia.

The primary amino group may be further alkylated and/or acylated to provide other intermediates of formula (III) wherein X²Restricted to CR⁵And R is⁵is-CH₂NR³R⁴。

Experimental method 26

According to scheme 26, wherein X²Restricted to CR⁵And R is⁵An intermediate of formula (XV) which is-CN, hereinafter referred to as an intermediate of formula (XL), may be prepared by the process wherein X²Restricted to CR⁵And R is⁵Is L^d(wherein L^dIs I or Br) (hereinafter referred to as intermediate of formula (XLI) by metal-mediated cyanation. Intermediates of formula (XLI) are commercially available or can be prepared according to conventional reaction methods generally known in the art. The cyanation reaction is usually carried out in a suitable reagent such as zinc cyanide (Zn (CN)₂) In the presence of oxygen. The reaction can be carried out in the presence of a catalyst such as tetrakis (triphenylphosphine) palladium (Pd (PPh)₃)₄) In the presence of a solvent such as DMF. Stirring and/or increasing the temperature (e.g., between 50-100 ℃) can increase the reaction rate.

Experimental method 27

Alternatively, intermediates of formula (IX) can also be prepared according to scheme 27 by reduction wherein R is defined as C_1-4Alkyl intermediates of formula (XLII), which are commercially available or can be prepared according to conventional reaction methods generally known in the art. This reaction can be carried out in the presence of a suitable reducing agent such as diisobutylaluminum hydride (DIBAL). This reaction can be carried out in a solvent such as DCM at low temperature (e.g., -78 ℃).

Experimental method 28

Intermediates of formula (XLII) wherein R is defined as C_1-4Alkyl groups, may be prepared by alkylation of intermediates of formula (XLIII) according to conventional reaction methods generally known in the art. According to scheme 28, the alkylation is carried out in the presence of a suitable base such as Cs₂CO₃Or K₂CO₃And alkylating agents, e.g. C_1-4In the presence of alkyl-Y (wherein Y is defined as Cl, Br or I). All other variables are as defined above. This reaction can be carried out in an aprotic solvent such as DMF.

Experimental method 29

Intermediates of formula (XLIII) can be prepared by oxidation of intermediates of formula (XLIV), which are commercially available or can be prepared according to conventional reaction methods generally known in the art. The oxidation is hydrogen peroxide (H) in a suitable oxidation system, for example trifluoroacetic anhydride (TFAA)₂O₂) In the presence of a solvent such as DCM or CH₃In CN.

Experimental method 30

According to scheme 30, intermediates of formula (XIII), wherein

-X¹Is limited to CH;

-X²is limited to CR^5aWherein R is^5aIs optionally each independently selected from C_1-4Alkoxy, fluoro, chloro and NR³R⁴C substituted by one or more substituents of_1-4Alkoxy or C_1-4An alkyl group;

-X³is limited to N;

-L is defined as Br, I or Cl;

hereinafter referred to as intermediates of formula (XLV), can be prepared by halogenation of intermediates of formula (XLVI) according to conventional reaction methods generally known in the art. The reaction is usually carried out in the presence of a halogenating agent such as phosphorus oxychloride in a solvent such as CH₃In CN. Stirring and/or increasing the temperature (e.g., between 50-100 ℃) can increase the reaction rate.

Experimental method 31

Intermediates of formula (XLVI) wherein R^5aAs defined in scheme 30, can be prepared by cyclization of an intermediate of formula (XLVII) according to conventional reaction methods generally known in the art. This reaction can generally be carried out at elevated temperature (above 220 ℃) in Dowtherm thermal Carrier (Dowtherm) A (biphenyl-diphenyl ether mixture).

Experimental method 32

Intermediates of formula (XLVII) can be prepared according to conventional reaction methods generally known in the art by condensation of intermediates of formula (XLVIII) with beta-ketoesters of formula (XLIX), wherein R is^5aAs defined in flow 30. This reaction can be carried out in a solvent such as benzene or toluene, usually in the presence of a catalytic amount of p-toluenesulfonic acid.

Experimental method 33

Intermediates of formula (XLVIII) can be prepared by conventional methods, such as reductive hydrogenation of intermediate (L).

Experimental method 34

The intermediate of formula (L) may be prepared by conventional methods, for example by nitration of the intermediate (L) of formula (LI) in a mixture of sulphuric and nitric acids.

Experimental method 35

According to scheme 35, intermediates of formula (II) wherein Het¹Limited to (a-2) shown in scheme 35, hereinafter referred to as intermediates of formula (LII), can also be prepared by copper catalyzed reaction of an intermediate of formula (LIII) with an (un) substituted imidazole or triazole of formula (XX), wherein halo is defined as Br or I and wherein all other variables are as defined above. The reaction can be carried out under a protective atmosphere such as N₂The process is carried out as follows. Agitation, elevated temperature (e.g., between 70-200 ℃) and/or increased pressure may increase the reaction rate. This reaction is usually carried out in an organic solvent such as DMSO or DMF. Optionally, the reaction may be carried out in a base such as K₂CO₃，Cs₂CO₃Or Et₃In the presence of N and/or ligands such as N, N' -dimethylethylenediamine or 1, 10-phenanthroline. As copper catalyst, catalytic or stoichiometric amounts of copper salts such as Cu may be used₂O, CuI or CuBr. By using compounds in accordance with standard practiceSuitable amino protecting groups, the amino groups in the intermediates of formula (LIII) may be protected before the reaction and the protection may be removed after the reaction, see for example t.w.greene and p.g.m.wuts in "protecting groups in Organic Chemistry", John Wiley and Sons, 1999.

Experimental method 36

Intermediates of formula (XXVII), wherein Het¹Limited to (a-2) and wherein G2 is specifically CH, as shown in scheme 36, hereinafter referred to as intermediates of formula (LIV), can be prepared by acylation of intermediate (LVIII) to give intermediate (LVII). According to scheme 36, the acylation reaction can be carried out in a reaction inert solvent such as THF, and optionally in a suitable base such as Et₃N or under acidic conditions, e.g. a mixture of acetic anhydride and formic acid. Subsequently, intermediates of formula (LV) can be prepared by alkylation of intermediates of formula (LVII) with intermediates of formula (LVI). The reaction can be carried out in a reaction-inert solvent such as DMF and a suitable base such as Cs₂CO₃Or K₂CO₃And optionally in the presence of catalytic amounts of an iodide salt such as KI or NaI. Subsequently, the intermediate (LV) is reacted with a source of ammonia such as ammonium acetate (NH)₄Condensation of OAc) gives the compound of formula (LIV). In scheme 36, halo is defined as Cl or Br, and all other variables are as defined above.

For the construction of the imidazole ring in the intermediate of formula (LIV), R is introduced¹⁰And R¹¹The order of (a) may be reversed. Reactions of this type are described in US2006/0004013 for 1- (4-bromo-2-methoxyphenyl) -4-methyl-1H-imidazole.

One or more of the following steps may be performed in any order, as desired or necessary:

the compounds of formula (I), any subgroup, addition salts, solvates, and stereochemically isomeric forms thereof, may be converted into other intermediates and compounds according to the present invention using methods known in the art.

One skilled in the art will appreciate that in the above-described methods, the functional groups of the intermediate compounds may need to be masked by protecting groups. In case the functional group of the intermediate compound is masked by a protecting group, it may be deprotected after the end of the reaction step.

Pharmacology of

The compounds of the invention have been found to modulate gamma-secretase activity. Thus, the compounds according to the present invention and pharmaceutically acceptable compositions thereof may be used for the treatment or prevention of AD, TBI, MCI, aging, dementia associated with lewy bodies, cerebrovascular amyloid diseases, multi-infarct dementia, down's syndrome, dementia associated with parkinson's disease and dementia associated with beta-amyloid, preferably AD.

As used herein, the term "modulating γ -secretase activity" refers to the effect on the treatment of APP by the γ -secretase-complex. Preferably it refers to a method wherein the overall rate of treatment of APP remains substantially the effect of no administration of the compound, but wherein the relative amount of product treated is altered, more preferably the production of a β 42-peptide is reduced. For example, different A.beta.species may be produced (e.g., A.beta.38 of shorter amino acid sequence or other A.beta.peptide species in place of A.beta.42) or the relative amounts of the products may be different (e.g., the ratio of A.beta.40 to A.beta.42 is altered, preferably increased).

It was previously demonstrated that the γ -secretase complex is also involved in handling Notch (Notch) -proteins. Notch is a signaling protein that plays a critical role in developmental processes (e.g., as reviewed in Schweisguth F (2004) curr. biol.14, R129). Regarding the use of gamma-secretase modulators in medicine, it appears to be particularly advantageous for notch-processing activities that do not interfere with gamma-secretase activity, in order to avoid the presumed unwanted side effects. Although gamma-secretase inhibitors exhibit side effects as a result of treatment accompanied by inhibition of notch, gamma-secretase modulators may have the following advantages: the selective reduction in production of a β in a highly aggregated and neurotoxic form, i.e., a β 42, is not accompanied by inhibition of notch processing, while the reduction in production of a β in a less aggregated form, i.e., a β 38, is not accompanied by inhibition of notch processing. Thus, preferred compounds are those that do not affect the notch-processing activity of the γ -secretase-complex.

As used herein, the term "treatment" means the entire process in which the progression of the disease can be slowed, interrupted, arrested or stopped, but need not indicate complete elimination of all symptoms.

The present invention relates to compounds according to general formula (I), the stereoisomeric forms thereof and the pharmaceutically acceptable acid or base addition salts and solvates thereof, for use as a medicament.

The invention also relates to compounds according to general formula (I), the stereoisomeric forms thereof and the pharmaceutically acceptable acid or base addition salts and solvates thereof, which are useful for modulating gamma-secretase activity.

The invention also relates to compounds according to general formula (I), the stereoisomeric forms thereof and the pharmaceutically acceptable acid or base addition salts and solvates thereof, for use in the treatment or prevention of a disease or disorder selected from AD, TBI, MCI, senility, dementia associated with lewy bodies, amyloid cerebrovascular disease, multi-infarct dementia or down syndrome.

In one embodiment, the disease or disorder is preferably alzheimer's disease.

The invention also relates to compounds according to general formula (I), the stereoisomeric forms thereof and the pharmaceutically acceptable acid or base addition salts and solvates thereof, which are useful for the treatment of said diseases.

The invention also relates to compounds according to general formula (I), the stereoisomeric forms thereof and the pharmaceutically acceptable acid or base addition salts and solvates thereof, for use in the treatment or prophylaxis, in particular in the treatment of γ -secretase mediated diseases or disorders.

The invention also relates to the use of a compound according to general formula (I), its stereoisomeric forms and the pharmaceutically acceptable acid or base addition salts and solvates thereof for the preparation of a medicament.

The invention also relates to the use of a compound according to general formula (I), its stereoisomeric forms and the pharmaceutically acceptable acid or base addition salts and solvates thereof for the preparation of a medicament for modulating the activity of gamma-secretase.

The invention also relates to the use of a compound according to general formula (I), the stereoisomeric forms thereof and the pharmaceutically acceptable acid or base addition salts and solvates thereof for the preparation of a medicament for the treatment or prevention of any one of the above mentioned disease conditions.

The invention also relates to the use of compounds according to general formula (I), the stereoisomeric forms thereof and the pharmaceutically acceptable acid or base addition salts and solvates thereof, for the preparation of a medicament for the treatment of any of the above mentioned disease states.

In the present invention, it is particularly preferred to administer a compound of formula (I) or any subgroup thereof which inhibits the IC production of the A β 42-peptide₅₀Values are less than 1000nM, preferably less than 100nM, more preferably less than 50nM, and even more preferably less than 20nM, when determined according to a suitable assay, such as the assay used in the examples below.

The compounds of the present invention may be administered to a mammal, preferably a human, for the treatment or prevention of any of the above-mentioned diseases.

In view of the use of the compounds of formula (I), there is provided a method of treating or preventing the development of any one of the above-mentioned diseases in warm-blooded animals, including humans.

Said method comprising the administration, that is to say systemic or topical administration, preferably oral administration, of an effective amount of a compound of formula (I), its stereoisomeric forms and pharmaceutically acceptable addition salts and solvates thereof, to warm-blooded animals including humans.

The invention also relates to the use of a compound of formula (I) for modulating gamma-secretase activity resulting in a reduction in the relative amount of a β 42-peptide produced.

An advantage of the compounds of the invention or a part of the compounds may be their enhanced CNS-penetration rate.

From the results presented below, a skilled artisan treating the disease can determine the daily amount of effective treatment. A therapeutically effective daily amount will be from about 0.005mg/kg to 50mg/kg, especially 0.01mg/kg to 50mg/kg body weight, more especially 0.01mg/kg to 25mg/kg body weight, preferably from about 0.01mg/kg to about 15mg/kg, more preferably from about 0.01mg/kg to about 10mg/kg, even more preferably from about 0.01mg/kg to about 1mg/kg, most preferably from about 0.05mg/kg to about 1mg/kg body weight. The amount of a compound according to the invention, also referred to herein as the active ingredient, required to achieve a therapeutic effect will, of course, vary from case to case as a function of the particular compound, the route of administration, the age and condition of the recipient, and the particular disorder or disease being treated.

The method of treatment may also include administering the active ingredient on a schedule between one and four intakes per day. In these methods of treatment, the compounds according to the invention are preferably formulated prior to administration. Suitable pharmaceutical formulations are prepared by known methods using well known and readily available ingredients, in accordance with the description herein below.

Compounds of the present invention suitable for treating or preventing Alzheimer's disease or symptoms thereof may be administered alone or in combination with one or more other therapeutic agents. Combination therapy includes administration of a single pharmaceutical dosage formulation containing a compound of formula (I) and one or more other therapeutic agents, as well as administration of a compound of formula (I) and each of the additional therapeutic agents under their respective separate pharmaceutical dosage formulations. For example, the compound of formula (I) and the therapeutic agent may be administered to the patient together in a single oral dosage composition, such as a tablet or capsule, or each agent may be administered in separate oral dosage formulations.

Although the active ingredient may be administered alone, it is preferably present as a pharmaceutical composition.

Accordingly, the present invention also provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and, as active ingredient, a therapeutically effective amount of a compound according to formula (I).

The carrier or diluent must be "acceptable" in the sense of being compatible with the other ingredients of the composition and not deleterious to the recipient thereof.

For ease of administration, the compounds of the present invention may be formulated in different pharmaceutical forms for administration purposes. The compounds according to the invention, in particular the compounds according to formula (I), the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof, or any subgroup or combination thereof, may be formulated into different pharmaceutical forms for administration purposes. Suitable compositions may refer to all compositions typically used for systemic administration of drugs.

To prepare the pharmaceutical compositions of this invention, an effective amount of the particular compound, optionally in addition salt form, as the active ingredient is intimately admixed with a pharmaceutically acceptable carrier, which may take a wide variety of forms depending on the form of preparation to be administered. These pharmaceutical compositions need to be in unit dosage form, suitable in particular for oral, rectal, subcutaneous, parenteral injection or inhalation administration. For example, in preparing the compositions in orally administrable form, any of the usual pharmaceutical media may be employed, such as water, glycols, oils, alcohols and the like, in the case of oral liquid preparations such as suspensions, syrups, elixirs, emulsions and solutions; in the case of powders, pills, capsules and tablets, solid carriers such as starches, sugars, kaolin, diluents, lubricants, binders, disintegrating agents and the like may be employed. Because of their ease of administration, tablets and capsules represent the most advantageous oral dosage unit form in which case solid pharmaceutical carriers are obviously employed. For parenteral compositions, the carrier will typically comprise sterile water, at least in large part, although other ingredients may be included, for example to aid solubility. For example, injectable solutions may be prepared in which the carrier comprises saline solution, glucose solution or a mixture of saline and glucose solution. For example, injectable solutions may be prepared in which the carrier comprises saline solution, glucose solution or a mixture of saline and glucose solution. Injectable solutions containing the compounds of formula (I) may be formulated in oils for prolonged action. Suitable oils for this purpose are, for example, peanut oil, sesame oil, cottonseed oil, corn oil, soybean oil, synthetic glycerol esters of long-chain fatty acids and mixtures of these with other oils. Suspensions which can be injected can also be prepared, in which case appropriate liquid carriers, suspending agents and the like can be used. Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations. In compositions suitable for transdermal administration, the carrier optionally contains penetration enhancers and/or suitable wetting agents, optionally in combination with small amounts of suitable additives of any nature that do not cause significant deleterious effects on the skin. The additives may facilitate administration to the skin and/or may aid in the preparation of the desired composition. These compositions can be administered in a variety of ways, for example as a transdermal patch, as a spot-on, as an ointment. Acid or base addition salts of the compounds of formula (I) are more suitable for the preparation of aqueous compositions because of their increased aqueous solubility over the corresponding base or acid form.

It is particularly advantageous to formulate the above-described pharmaceutical compositions in unit dosage form to provide ease of administration and uniformity of dosage. Unit dosage form as used herein means physically discrete units suitable as unitary dosages, each unit containing a predetermined quantity of active ingredient calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. Examples of such unit dosage forms are tablets (including scored or coated tablets), capsules, pills, powder packets, wafers, suppositories, injectable solutions or suspensions and the like, and segregated multiples thereof.

Since the compounds according to the invention are effective orally administrable compounds, orally administrable pharmaceutical compositions containing said compounds are particularly advantageous.

In order to increase the solubility and/or stability of the compounds of formula (I) in the pharmaceutical composition, it may be advantageous to use α -, β -or γ -cyclodextrins or derivatives thereof, in particular hydroxyalkyl-substituted cyclodextrins, such as 2-hydroxypropyl- β -cyclodextrin or sulfobutyl- β -cyclodextrin. Auxiliary solvents such as alcohols may also improve the solubility and/or stability of the compounds according to the invention in the pharmaceutical composition.

Depending on the mode of administration, the pharmaceutical composition will preferably contain from 0.05 to 99% by weight, more preferably from 0.1 to 70% by weight, even more preferably from 0.1 to 50% by weight of a compound of formula (I), and from 1 to 99.95% by weight, more preferably from 30 to 99.9% by weight, even more preferably from 50 to 99.9% by weight of a pharmaceutically acceptable carrier, all percentages being based on the total weight of the composition.

The following examples illustrate the invention.

Examples

Hereinafter, the term "DCM" refers to dichloromethane; "MeOH" refers to methanol; "HPLC" refers to high performance liquid chromatography; "sat." means saturated; "aq." refers to an aqueous solution; "r.t." means room temperature; "AcOH" refers to acetic acid; "RP" refers to the reverse phase; "min" means minutes; "h" means hours; "i.d." means inside diameter; "EtOAc" refers to ethyl acetate; "NaOAc" refers to sodium acetate; "KOtBu" refers to potassium tert-butoxide; "Et 3N" refers to triethylamine; "EtOH" refers to ethanol; "eq" refers to equivalent; "r.m." means the reaction mixture; "DIPE" refers to diisopropyl ether; "THF" refers to tetrahydrofuran; "DME" refers to dimethoxyethane(ii) a "DMSO" refers to dimethylsulfoxide; "BINAP" refers to [1, 1' -binaphthyl]-2, 2' -diylbis [ diphenylphosphine](racemization); "NH₄OAc "refers to ammonium acetate; "DMF" refers to N, N-dimethylformamide; "X-Phos" refers to dicyclohexyl [2 ', 4', 6 '-tris (1-methylethyl) [1, 1' -biphenyl ]]-2-yl]A phosphine; and is "Pd₂(dba)₃"refers to tris [ mu- [ (1, 2-eta: 4, 5-eta) - (1E, 4E) -1, 5-diphenyl-1, 4-pentadien-3-one]]Dipalladium.

A. Preparation of intermediates

Example A1

a) Preparation of intermediate 1

In N₂1-chloro-2-methoxy-4-nitrobenzene (50g, 0.26mol), 4-methyl-1H-imidazole (43.77g, 0.53mol) and K were reacted under an atmosphere of nitrogen₂CO₃A mixture (36.84g, 0.26mol) in DMSO (500ml) was reacted in an autoclave at 150 ℃ for 6 hours. This reaction was repeated twice (150 g in total) with 50g of 1-chloro-2-methoxy-4-nitrobenzene each. Subsequently, 3 reaction mixtures were combined and poured into ice-water (6 l). The solid is filtered off and taken up in H₂And O washing. Dissolve the solid in DCM and take the solution with H₂And O washing. The separated organic layer was dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified over silica gel on a glass filter (eluent: DCM/MeOH from 100/0 to 97/3). The product fractions were collected and the solvent was evaporated. The residue was suspended in DIPE, filtered and dried in an oven. Yield: 48.54g of intermediate 1 (26%).

b) Preparation of intermediate 2a and intermediate 2

Intermediate 1(13.2g, 56.6mmol) was dissolved in MeOH (250 ml). Pd/C (0.5g) was added to the solution and the resulting suspension was brought to 50 ℃ and H₂Stirring (atmospheric pressure) overnight. Consumption H₂After (1 eq.) the catalyst was filtered off. The organic layer was evaporated to give intermediate 2a (free base). Intermediate 2a was dissolved in HCl/EtOH solution and stirred for 30 minutes. The solvent was removed in vacuo. The residue was crystallized from EtOH containing a small amount of petroleum ether to give the desired product. Yield: 4.7g of intermediate 2 (41.0%;. HCl).

Example A2

a) Preparation of intermediates 3 and 4

1-fluoro-2-methoxy-4-nitrobenzene (821mg, 4.8mmol), 5-methyl-1H-1, 2, 4-triazole (800mg, 9.63mmol), K₂CO₃A mixture of (4.8mmol) and DMSO (8ml) was stirred at 120 ℃ for 1 hour. After cooling, the reaction mixture was poured into ice water. The solid is filtered off, washed with water and dried (vacuum; 50 ℃). Yield: 0.554g of intermediate 3 (49%). The aqueous layer was saturated with NaCl, extracted with DCM and the organic layer dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography on silica gel (eluent: DCM). The desired fractions were collected and the solvent was evaporated. Yield: 0.147g of intermediate 4 (13%).

b) Preparation of intermediate 5

In N₂MeOH (50ml) was added to Pd/C10% (150mg) under atmosphere. Subsequently, a 0.4% thiophene solution in DIPE (1ml) and intermediate 3(550mg, 2.348mmol) were added. The reaction mixture was heated at 25 ℃ and N₂Stirred under atmosphere to 3 equivalents of H₂Is absorbed. The catalyst was filtered through celite. The filtrate was evaporated and the residue was suspended in DIPE, filtered and dried in vacuo. Yield: 0.350g of intermediate 5 (73.0%).

Example A3

a) Preparation of intermediate 6

Will K₂CO₃(9.6g, 69.5mmol) and 1-methyl-1-tosylmethylisonitrile (8g, 38.2mmol) were added to a solution of 2-formyl-5-nitroanisole (6.29g, 34.7mmol) in MeOH (150ml) and the reaction mixture was refluxed for 4 h. The reaction mixture was concentrated under reduced pressure, the residue was dissolved in DCM and the organic phase was taken up with H₂O washing and drying (MgSO)₄) Filtered and the solvent evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluent: n-heptane/EtOAc from 100/0 to 50/50). The product fractions were collected and the solvent was evaporated. Yield: 6.24g of intermediate 6 (77%).

b) Preparation of intermediate 7

In N₂MeOH (150ml) was added to Pd/C10% (1g) under an atmosphere. Subsequently, a 0.4% thiophene solution in DIPE (1ml) and intermediate 6(6.24g, 26.6mmol) were added. H of the reaction mixture at 25 ℃₂Stirred under atmosphere to 3 equivalents of H₂Is absorbed. The catalyst was filtered through celite and the filtrate was evaporated. Yield: 5.4g intermediate 7 (99%).

Example A4

a) Preparation of intermediate 8

At room temperature and N₂Iodobenzene diacetate (2.47g, 7.68mmol) and trifluoromethanesulfonic acid (1.35ml, 15.3mmol) in CH under an atmosphere₃CN (40ml) for 1 hour. Subsequently, the mixture was heated to reflux temperature. 2 '-methoxy-4' -nitro-acetophenone (1.0g, 5.12mmol) was added all at once to the solution and the reaction mixture was refluxed for 2 hours, then cooled to room temperature and the solvent was evaporated. The residue was partitioned between saturated aqueous sodium bicarbonate (200ml) and EtOAc (200 ml). The organic layer was separated and washed with brine and dried (MgSO₄) Filtered and evaporated to give a brown solid. The product was purified by flash chromatography on silica gel (eluent: DCM/MeOH from 100/0 to 99/1). The product fractions were collected and the solvent was evaporated (reduced pressure). Yield: 0.42g of intermediate 8 (35%).

b) Preparation of intermediate 9

In N₂MeOH (50ml) was added to Pd/C10% (0.250g) under atmosphere. Subsequently, a 0.4% thiophene solution in DIPE (2ml) and intermediate 8(0.946g, 4.04mmol) were added. At 25 ℃ H₂The reaction mixture was stirred under an atmosphere to 3 equivalents of H₂Is absorbed. The catalyst was filtered through celite and the filtrate was evaporated. The product was triturated in DIPE, filtered off and dried in vacuo. Yield: 0.66g intermediate 9 (80%).

Example A5

a) Preparation of intermediate 10

Will K successively₂CO₃(36g, 262mmol) and 1-methyl-1-tosylmethylisonitrile (35g, 167mmol) were added to a solution of 5-nitropyridine-2-aldehyde (20g, 131mmol) in MeOH (500ml) and the reaction mixture was refluxed for 4 h. The reaction mixture was concentrated under reduced pressure, the residue was dissolved in DCM and the organic phase was taken up with H₂O washing and drying (Na)₂SO₄) Filtered and the solvent evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluent: petroleum ether/EtOAc 4/1). The product fractions were collected and the solvent was evaporated. Yield: 15g of intermediate 10 (56%).

b) Preparation of intermediate 11

A solution of intermediate 10(10g, 48.7mmol) in THF (300ml) was added NH₄Cl (2.6g, 48.7mmol) in H₂O (100 ml). Iron (16.3g, 292mmol) was added and the reaction mixture was refluxed for 4 hours. The precipitate was removed by filtration and the filtrate was evaporated in vacuo. The residue was dissolved in EtOAc and the organic layer was washed with H₂O washing and drying (Na)₂SO₄) Filtered and the solvent evaporated in vacuo. The residue was dissolved in 2N HCl solution and the aqueous layer was washed with DCM, made basic by addition of 2N NaOH and the product was extracted with EtOAc. The organic layer was dried (Na)₂SO₄) Filtration and evaporation of the solvent in vacuo, yield: 6g of intermediate 11 (71%).

Example A6

a) Preparation of intermediate 12

2-iodo-5-bromopyridine (13.7g, 48.2mmol), 2-methyl-4-Methyl Azolodicarboxylate (3.4g, 24.1mmol), Palladium (II) acetate (0.54g, 2.41mmol), Tri-o-tolylphosphine (1.47g, 4.81mmol) and Cs₂CO₃A solution of (15.7g, 48.2mmol) in toluene (75ml) was treated with N₂Purged, sealed and stirred at 110 ℃ overnight. The catalyst was filtered through celite and the filtrate was evaporated. The crude product was purified by flash chromatography on silica gel (eluent: DCM/MeOH (NH)₃) From 100/0 to 98/2). The product fractions were collected and the solvent was evaporated. Yield: 5.64g of intermediate 12 (64%).

b) Preparation of intermediate 13

Intermediate 12(5.64g, 15.4mmol) and LiOH (0.91g, 38mmol) were dissolved in diAlkane (40ml) and H₂O (10 ml). The reaction mixture was stirred at room temperature for 5 hours and then treated with 1M HCl solution to pH 2. The precipitate was filtered and dried under vacuum. Filtrate with CHCl₃Extract and dry the organic layer (MgSO)₄) Solid was obtained after filtration and removal of the solvent under reduced pressure. The two solid fractions were combined. Yield: 4.75g of intermediate 13 (97%).

c) Preparation of intermediate 14

A solution of intermediate 13(3.3g, 11.65mmol) in a mixture of DMF (75ml) and EtOH (30ml) was heated under microwave conditions at 150 ℃ for 4 hours. After cooling, the solvent was evaporated to yield intermediate 14(3.1g, 89%). This fraction was used in the next step without further purification.

d) Preparation of intermediate 15

Reacting [ (R) -1- [ (S) -2- (dicyclohexylphosphino) ferrocenyl]Ethyl radical]Di-tert-butylphosphine (Josi-Phos, 0.492g, 0.89mmol) and Pd (OAc)₂After premixing in DME (2ml) a solution of intermediate 14(4.25g, 17.8mmol) and sodium tert-butoxide (2.39g, 6.69mmol) in DME (18ml) was added. Finally, N-benzylamine (2.28g, 21.33mmol) was added and the reaction mixture was stirred at 100 ℃ for 9 hours. After cooling, the reaction mixture was diluted with DCM and filtered through celite. The filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: DCM/MeOH (NH)₃) From 100/0 to 98/2). The product fractions were collected and the solvent was evaporated under reduced pressure. Yield: 3.23g of intermediate 15 (67%).

e) Preparation of intermediate 16

In N₂MeOH (50ml) was added to Pd/C10% (0.05g) under atmosphere. Intermediate 15(0.15g, 0565mmol) was added and heated at 50 ℃ under H₂The reaction mixture was stirred to 1 equivalent of H under an atmosphere₂Is absorbed. The catalyst was filtered through celite. The filtrate was evaporated. Yield: 0.105g intermediate 16 (95%).

Example A7

a) Preparation of intermediate 17

1-methyl-4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyrazole (2.83g, 13.63mmol), CsF (3.11g, 20.45mmol) and [1, 1' -bis (diphenylphosphino) ferrocene]Palladium (II) dichloride (1.99g, 13.64mmol) was added to a solution of 4-bromo-3-fluoronitrobenzene (3.0g, 4.81mmol) in DMF (60 ml). The reaction mixture is treated with N₂Purged, sealed and stirred at 100 ℃ for 8 hours. After cooling, the solvent was evaporated. The residue was dissolved in DCM and the organic phase was washed with H₂O washing and drying (MgSO)₄) Filtration and concentration of the filtrate under reduced pressure gave intermediate 17. This fraction was used in the next step without further purification.

b) Preparation of intermediate 18

Intermediate 17(3.0g, 13.56mmol) and iron (3.78g, 67.8mmol) were shaken in AcOH (24ml) for 1.5 h. The solvent was evaporated. The residue was dissolved in DCM and the organic layer was washed with saturated Na₂CO₃The solution was washed and dried (MgSO)₄) Filtered and concentrated under reduced pressure. The residue was triturated in DIPE and the resulting precipitate was filtered. Yield: 0.72g of intermediate 18 (28%).

Example A8

a) Preparation of intermediate 19

3-bromo-2-toluene (10.0g, 42.29mmol), dimethylformamide dimethyl acetal (15.55g, 139mmol) and pyrrolidine (3.29g, 46.29mmol) were stirred at 115 ℃ for 22 h. The solution was cooled to room temperature and used as such in the next step.

b) Preparation of intermediate 20

The crude solution from the previous reaction step containing intermediate 19 was added dropwise sodium periodate (29.7g, 139mmol) in DMF (75ml) and H at 0 deg.C₂O (100ml) in a stirred solution. The reaction mixture was then allowed to warm to room temperature and stirred for 3 hours. The suspension was filtered through celite, which was washed thoroughly with EtOAc. The filtrate is treated with H₂O wash and concentrate the organic phase under reduced pressure. The residue was purified by chromatography on silica gel (eluent: n-heptane/DCM from 50/50 to 0/100). The product fractions were collected and the solvent was evaporated. Yield: 2.72g of intermediate 20 (20% yield over 2 reaction steps).

c) Preparation of intermediate 21

Sodium triacetoxyborohydride (1.38g, 6.5mmol) was added dropwise to a stirred solution of intermediate 20(1.0g, 4.34mmol), 3-methoxyaniline (0.53g, 4.34mmol) and acetic acid (1.3g, 21.7mmol) in 1, 2-dichloroethane (16 ml). The reaction mixture was stirred at room temperature for 4 hours with K₂CO₃Aqueous and brine washes. The organic phase was dried (MgSO)₄) Filtered and the solvent removed under reduced pressure. The residue was purified by flash chromatography on silica gel (eluent: n-heptane/DCM, equivalent 50/50). The product fractions were collected and the solvent was evaporated. Yield: 0.65g of intermediate 21 (41%).

d) Preparation of intermediate 22

A mixture of intermediate 21(5.68g, 16.8mmol) and tin (II) chloride dihydrate (7.6g, 33.7mmol) in EtOH (100ml) was stirred at 40 ℃ overnight. The solvent was evaporated and the residue was suspended in H₂O, and the product was extracted thoroughly with DCM. The organic phase was dried (MgSO)₄) Filtered and the solvent removed (reduced pressure). The residue was purified by chromatography on silica gel (eluent: n-heptane/DCM from 40/60 to 0/100). The product fractions were collected and the solvent was evaporated. Yield: 3.63g of intermediate 22 (71%).

e) Preparation of intermediate 23

A2M solution of lithium diisopropylamide in THF was added dropwise to a-78 deg.C solution of intermediate 22(3.0g, 9.9mmol) in THF. The reaction mixture was warmed to 0-5 ℃ and stirred for 15 minutes. The mixture was cooled again to-78 ℃ and CH was added₃I (2.1g, 14.8 mmol). The temperature of the reaction mixture was slowly raised to room temperature and stirred for 16 hours. Addition of H₂O and the product is extracted with diethyl ether. The organic layer was dried (MgSO₄) Filtered and the solvent removed under reduced pressure. The residue was purified by chromatography on silica gel (eluent: n-heptane/DCM from 50/50 to 0/100). The product fractions were collected and the solvent was evaporated. Yield: 3.63g of intermediate 23 (71%).

Example A9

a) Preparation of intermediate 24

2-bromo-6-methylaniline (1.18g, 6.34mmol) was dissolved in 6N aqueous HClAfter stirring at 60 ℃ for 30 minutes, the reaction mixture was cooled to 0 ℃. Dropwise adding NaNO₂(0.481g, 6.98mmol) in H₂O (1.5ml) and the reaction mixture was stirred at 0 ℃ for a further 1 h. The reaction mixture was buffered (pH between 4 and 5) by addition of NaOAc saturated aqueous solution and the mixture was then added in portions to an ice-cold solution of tert-butylmercaptan 0.63g, 6.98mmol) in EtOH (25 ml). The reaction mixture was allowed to warm to room temperature and stirred overnight. The reaction mixture was partitioned between EtOAc (100mL) and H₂O (100 ml). The aqueous layer was extracted with EtOAc and the combined organic layers were dried (MgSO)₄) Filtered and the solvent removed under reduced pressure. Yield: 1.7g of intermediate 24 (70%).

b) Preparation of intermediate 25

A solution of intermediate 24(1.7g, 4.44mmol) in DMSO (20ml) was added dropwise to a solution of KOtBu (6.64g, 59mmol) in DMSO (50 ml). The reaction mixture was stirred at room temperature for 2 hours and then poured onto ice (300g) containing 1N aqueous HCl (300 ml). The mixture was extracted with diethyl ether. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent removed under reduced pressure. The residue was purified by chromatography on silica gel (eluent: DCM). The product fractions were collected and the solvent was evaporated. Yield: 0.55g of intermediate 25 (63%).

c) Preparation of intermediate 26

A mixture of intermediate 25(0.54g, 2.74mmol) and dibutyl sulfate (0.493g, 2.77mmol) in toluene (7ml) was stirred at 110 ℃ for 24 h. The reaction mixture was cooled to room temperature and quenched with NaHCO₃Washing with saturated aqueous solution. The organic phase was dried (MgSO)₄) Filtered and the solvent evaporated. The crude oil was purified by chromatography on silica gel (eluent: n-heptane/DCM from 90/10 to 70/30). The product fractions were collected and the solvent was evaporated. Yield: 0.335g of intermediate 26 (43%).

Example A12

Preparation of intermediate 27

Intermediate 25(2g, 10.1mmol), perfluorobutanesulfonic acid 2, 2, 2-trifluoroethyl ester (4.9g, 12.84mmol) and Cs₂CO₃(9.92g, 30.45mmol) of the solution was stirred at room temperature for 4 hours. The reaction mixture was diluted with EtOAc and washed with H₂And O washing. The organic layer was dried (MgSO₄) Filtered and the solvent evaporated. The resulting yellow oil was purified by chromatography on silica gel (eluent: n-heptane/DCM from 80/20 to 0/100). The product fractions were collected and the solvent was evaporated. Yield: 1.08g of intermediate 27 (38%).

Example A13

a) Preparation of intermediate 28

Pyridine chlorochromate(67g, 310mmol) was added to a suspension of 3-chloro-2-nitrobenzyl alcohol (25g, 129mmol), molecular sieves (40g) and celite (40g) in DCM (500 ml). The reaction mixture was stirred at room temperature for 2 hours and then filtered through silica gel (eluent: DCM). The product fractions were collected and the solvent was removed under reduced pressure. The residue was purified by chromatography on silica gel (eluent: DCM). The product fractions were collected and the solvent was evaporated. Yield: 22.5g of intermediate 28 (94%))。

b) Preparation of intermediate 29

4-fluoroaniline (1.83g, 16.1mmol) was added dropwise over 10 min to a solution of intermediate 28(3g, 16.1mmol) in AcOH (50 ml). Trimethylsilyl cyanide (4.3ml, 32.3mmol) was added dropwise and the reaction mixture was stirred at room temperature for 16 h. Evaporate the solvent and partition the residue on H₂Between O and DCM. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent removed under reduced pressure. The residue was dissolved in EtOH (100ml) under mild heating and 0.5M Na was added₂CO₃Solution (15ml, 0.75 mmol). The bright yellow indazole oxide begins to crystallize almost immediately. The mixture was allowed to cool to room temperature. The precipitate was filtered and recrystallized from EtOH/AcOH. Yield: 1.9g of intermediate 29 (40%).

c) Preparation of intermediate 30

Phosphorus trichloride (4.72g, 34.3mmol) was added to intermediate 29(1.6g, 5.56mmol) in CHCl₃(25ml) and the reaction mixture was refluxed for 1 hour. After cooling, the reaction mixture was poured into ice-water. The aqueous layer was basified (NaOH) and the product was extracted with DCM. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent removed under reduced pressure. Removing the residue from CH₃CN crystallized, filtered and dried in vacuo. Yield: 0.92g of intermediate 30 (61%).

Example A14

a) Preparation of intermediate 31

A mixture of formic acid (12.8ml, 340mmol) and acetic anhydride (8.54ml, 91mmol) was stirred at room temperature for 40 minutes. Subsequently, a solution of 3-amino-6-bromo-2-methoxy-pyridine (5g, 24.6mmol) in THF (30ml) was added dropwise to the mixture. The resulting reaction mixture was stirred at 60 ℃ overnight, then cooled and poured into ice-water, resulting in a solid precipitate. The solid was filtered, washed with water, and dried. Yield: 5.2g of intermediate 31 (76%).

b) Preparation of intermediate 32

1-chloro-propan-2-one (4.34g, 46.9mmol) was added dropwise to intermediate 31(5.2g, 18.8mmol), KI (0.343g, 2.06mmol) and Cs₂CO₃(21.4g, 65.9mmol) in a mixture of DMF (50 ml). The reaction mixture was stirred at room temperature overnight. Subsequently, the reaction mixture was poured into ice-water and extracted with EtOAc. The combined organic layers were dried (MgSO)₄) Filtered and concentrated in vacuo. The residue was suspended in DIPE and the resulting solid was filtered, washed with DIPE and dried. Yield: 443g of intermediate 32 (82%).

c) Preparation of intermediate 33

Intermediate 32(4.4g, 15.3mmol) was added to NH₄OAc (5.41g, 70.2mmol) in a mixture of AcOH (10 ml). The reaction mixture was refluxed for 1 hour. The reaction mixture was cooled to room temperature and poured into a mixture of ice-water and EtOAc. The mixture was basified to pH 9 with 50% w/v aqueous NaOH. Separating the organic layer, and dryingDried (MgSO)₄) Filtered and concentrated in vacuo. The solid product obtained was used as such in the next step. Yield: 3.78g of intermediate 33.

d) Preparation of intermediate 34

2-methyl-2-propanol sodium salt (0.717g, 7.46mmol), BINAP (464mg, 0.746mmol), Pd₂(dba)₃A mixture of (342mg, 0.373mmol), intermediate 33(1.0g, 3.73mmol) and benzophenone imine (0.845g, 4.66mmol) in toluene (20 ml; pre-deoxygenation) was stirred and heated under microwave conditions at 100 ℃ for 2 hours. The mixture was cooled and the solvent was removed in vacuo. THF (50ml) and 1N aqueous HCl (50ml) were added to the residue, and the mixture was stirred at room temperature for 1 hour. With 10% Na₂CO₃The reaction mixture was basified with aqueous solution and extracted with EtOAc. The organic layer was dried (MgSO₄) Filtered and the solvent evaporated in vacuo. The product was purified by flash chromatography on silica gel (eluent: DCM/MeOH from 100/0 to 95/5). The product fractions were collected and the solvent was evaporated. Yield: 0.6g of intermediate 34 (52% over 2 reaction steps).

Example A15

a) Preparation of intermediate 35

Sodium triacetoxyborohydride (1.17g, 5.5mmol) was added dropwise to a stirred solution of intermediate 20(0.8g, 3.69mmol), 4-fluorobenzylamine (0.46g, 3.69mmol) and AcOH (1.1g, 18.48mmol) in 1, 2-dichloroethane (12 ml). The reaction mixture was stirred at room temperature for 4 hours with K₂CO₃Aqueous and brine washes. The organic phase was dried (MgSO)₄) Is filtered and mixed inThe solvent was removed under reduced pressure. The residue was purified by flash chromatography on silica gel (eluent: n-heptane/DCM from 30/70 to 0/100). The product fractions were collected and the solvent was evaporated. Yield: 0.70g of intermediate 35 (41%).

b) Preparation of intermediate 36

A mixture of intermediate 35(0.6g, 1.77mmol) and tin (II) chloride dihydrate (0.80g, 3.59mmol) in EtOH (15ml) was stirred at 40 ℃ overnight. Evaporate the solvent and suspend the residue in H₂O, and the product was extracted thoroughly with DCM. Drying (MgSO)₄) The organic phase was filtered and the solvent was removed (reduced pressure). The residue was purified by RP preparative HPLC [ RPShandon Hyperprep ]C18 BDS (8 microns, 250g, i.d.5 cm); mobile phase: gradient (0.25% NH in water)₄HCO₃solution)/MeOH/CH₃CN]. The product fractions were collected and the solvent was evaporated. Yield: 0.094g intermediate 36 (17%).

B. Preparation of the Compounds

Example B1

Preparation of Compound 1

Intermediate 22(0.28g, 0.92mmol), Pd₂(dba)₃(0.084g, 0.092mmol), X-Phos (0.097g, 0.203mmol) and Cs₂CO₃(0.90g, 2.77mmol) was added to a solution of intermediate 2a (0.187g, 0.92mmol) in 2-methyl-2-propanol (10 ml). The reaction mixture was heated at 110 ℃ for 20 hours. After cooling, H is added₂O and the product was extracted with DCM. The organic phase was dried (MgSO)₄) Filtered and concentrated under reduced pressure. The residue was purified by RP preparative HPLC [ RP Shandon Hyperprep ]C18 BDS (8 microns, 250g, i.d.5 cm); mobile phase: (0.25% NH in Water₄HCO₃Solution of CH₃CN)]. The product fractions were collected and concentrated under reduced pressure. Yield: 0.156g of Compound 1 (40%).

Example B2

Preparation of Compound 2

Intermediate 23(0.152g, 0.48mmol), Pd₂(dba)₃(0.044g, 0.048mmol), X-Phos (0.050g, 0.105mmol) and Cs₂CO₃(0.47g, 1.43mmol) was added to a solution of intermediate 2a (0.097g, 0.48mmol) in 2-methyl-2-propanol (10 ml). The reaction mixture was heated at 110 ℃ for 20 hours. After cooling, H is added₂O and the product was extracted with DCM. The organic phase was dried (MgSO)₄) Filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: DCM/MeOH from 100/0 to 95/5) and the product fractions were collected and worked up. The residue was crystallized from DIPE, filtered and dried in vacuo at 60 ℃. Yield: 0.131g of Compound 2 (62%).

Example B3

Preparation of Compound 3

Intermediate 26(0.10g, 0.39mmol), Pd₂(dba)₃(0.036g，0.039mmol), X-Phos (0.041g, 0.087mmol) and Cs₂CO₃(0.38g, 1.18mmol) was added to a solution of intermediate 2a (0.080g, 0.39mmol) in 2-methyl-2-propanol (7 ml). The reaction mixture was heated at 110 ℃ for 20 hours. After cooling, H is added₂O and the product was extracted with DCM. The organic phase was dried (MgSO)₄) Filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: DCM/MeOH from 100/0 to 96/4) and the product fractions were collected and worked up to give crude compound 3a (free base of compound 3). The product was dissolved in DIPE and converted into its HCl-salt by adding 1ml of 6N HCl solution in 2-propanol, filtered off and dried under vacuum at 60 ℃. Yield: 0.070g of Compound 3 (39%;. 2 HCl).

Example B4

Preparation of Compound 4

Intermediate 23(0.317g, 1mmol), Pd₂(dba)₃(0.091g, 0.1mmol), X-Phos (0.095g, 0.2mmol) and Cs₂CO₃(0.98g, 3mmol) was added to a solution of intermediate 11(0.175g, 1mmol) in 2-methyl-2-propanol (10ml) and the reaction mixture was heated at 110 ℃ for 14 h. After cooling, H is added₂O and the product was diluted with DCM and filtered through celite. The filtrate is treated with H₂O washing and drying (MgSO)₄) Filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: DCM/MeOH (NH)₃) From 100/0 to 98/2). The product fractions were collected and the solvent was evaporated. Yield: 0.198g of Compound 4 (48%).

Example B5

Preparation of Compound 5

Intermediate 23(0.348g, 1.1mmol), Pd₂(dba)₃(0.091g, 0.1mmol), X-Phos (0.105g, 0.22mmol) and Cs₂CO₃(0.98g, 3mmol) was added to a solution of intermediate 5(0.204g, 1mmol) in 2-methyl-2-propanol (12ml) and the reaction mixture was heated at 110 ℃ for 20 h. After cooling, H is added₂O and the product was extracted with DCM. The organic phase was dried (MgSO)₄) Filtered and concentrated under reduced pressure. The residue was purified by column chromatography over silica gel (eluent: DCM/MeOH from 100/0 to 95/5). The product fractions were collected and worked up to give crude compound 5a (free base of compound 5). The product was dissolved in DIPE and converted into its HCl-salt by addition of 2ml of 6N HCl solution in 2-propanol, filtered off and dried under vacuum at 60 ℃. Yield: 0.344g of Compound 5 (67%; 1.9 HCl).

Example B6

Preparation of Compound 6

Intermediate 27(0.204g, 0.73mmol), Pd₂(dba)₃(0.064g, 0.07mmol), X-Phos (0.073g, 0.15mmol) and Cs₂CO₃(0.68g, 2.1mmol) was added to a solution of intermediate 9(0.142g, 0.7mmol) in 2-methyl-2-propanol (10ml) and the reaction mixture was heated at 60 ℃ for 16 h. After cooling, H is added₂The reaction mixture was diluted with DCM and filtered through celite. The filtrate is treated with H₂O washing and drying (MgSO)₄) Filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: DCM/MeOH (NH)₃) From 100/0 to 99/1). The product fractions were collected and the solvent was evaporated. Yield: 0.081g of Compound 6 (29%).

Example B7

Preparation of Compound 7

Intermediate 23(0.222g, 0.7mmol), Pd₂(dba)₃(0.064g, 0.07mmol), X-Phos (0.073g, 0.15mmol) and Cs₂CO₃(0.684g, 2.1mmol) intermediate 18(0.175g, 1mmol) in 2-methyl-2-propanol (12ml) was added and the reaction mixture was heated at 100 ℃ for 20 h. After cooling, H is added₂O and the product was diluted with DCM and filtered through celite. H for filtrate₂O washing and drying (MgSO)₄) Filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: DCM/MeOH (NH)₃) From 100/0 to 98/2). The product fractions were collected and the solvent was evaporated. Yield: 0.054g Compound 7 (18%).

Example B8

a) Preparation of Compound 32

Intermediate 30(0.198g, 0.73mmol), Pd₂(dba)₃(0.066g, 0.073mmol), X-Phos (0.076g, 0.16mmol) and Cs₂CO₃(0.714g, 2.2mmol) 4- (2-methyl-1, 3-)Oxazol-5-yl) aniline (0.127g, 0.73mmol) in 2-methyl-2-propanol (12ml) and the reaction mixture was heated at 110 ℃ for 20 h. After cooling, H is added₂O and the product was extracted with DCM. The organic phase was dried (MgSO)₄) Filtered and concentrated under reduced pressure. The residue was purified by column chromatography over silica gel (eluent: DCM/MeOH from 100/0 to 98/2). The product fractions were collected and worked up. The product is separated from CH₃CN, filtering and drying (In vacuum; 60 ℃ C.). Yield: 0.098g of Compound 32 (33%).

Preparation of Compound 8

In N₂MeOH/NH under atmosphere₃(40ml) was added to Raney nickel (0.05 g). Subsequently, compound 32(0.042g, 0.10mmol) was added. The reaction mixture was heated at 14 ℃ and H₂Stirred under atmosphere to 2 equivalents of H₂Is absorbed. The catalyst was filtered through celite and the filtrate was evaporated. The residue was purified by flash chromatography on silica gel (eluent: DCM/MeOH (NH)₃)95/5). The product fractions were collected and the solvent was evaporated. Yield: 0.010g of Compound 8 (23%).

Example B9

Preparation of Compound 9

Intermediate 27(0.278g, 0.99mmol), Pd₂(dba)₃(0.083g, 0.09mmol), X-Phos (0.095g, 0.2mmol) and Cs₂CO₃(0.885g, 2.72mmol) was added to a solution of intermediate 34(0.185g, 0.91mmol) in 2-methyl-2-propanol (10ml) and the reaction mixture was heated at 70 ℃ for 16 h. After cooling, H is added₂The reaction mixture was diluted with DCM and filtered through celite. The filtrate is treated with H₂O washing and drying (MgSO)₄) Filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: DCM/MeOH (NH)₃) From 100/0 to 98/2). The product fractions were collected and the solvent was evaporated. Yield: 0.092g of Compound 9 (25%).

Example B10

Preparation of Compound 10

Intermediate 36(0.094g, 0.308mmol), Pd₂(dba)₃(0.028g, 0.031mmol), X-Phos (0.032g, 0.068mmol) and Cs₂CO₃(0.301g, 0.92mmol) was added to a solution of intermediate 2a (0.062g, 0.308mmol) in 2-methyl-2-propanol (5ml) and the reaction mixture was heated at 110 ℃ for 20 h. After cooling, H is added₂O and the product was extracted with DCM. The organic layer was dried (MgSO₄) Filtered and concentrated under reduced pressure. The residue was purified by column chromatography over silica gel (eluent: DCM/MeOH from 100/0 to 98/2). The product fractions were collected and concentrated under reduced pressure. Yield: 0.070g of Compound 10 (53%).

Compounds 1 to 57 in tables 1a and Ib were prepared analogously to one of the above examples. If no salt formation is indicated, the compound is obtained as the free base. 'Pr.' refers to the example number of the compound synthesized according to this scheme. No.' refers to compound number.

To obtain the HCl salt form, several methods known to those skilled in the art are used. In a typical procedure, for example, the crude residue (free base) is dissolved in DIPE or Et₂O, and then dropwise added to a 6N HCl solution in 2-propanol or in Et₂1N HCl solution in O. The mixture was stirred for 10 minutes and the product was filtered. The HCl salt was dried in vacuo.

TABLE 1a

TABLE 1b

Analysis section

LCMS (liquid chromatography/mass spectrometry)

General procedure A

LC measurements were performed using an acquisition UPLC (ultra performance Liquid Chromatography) (Waters) system including a binary pump, a sample conditioner (samplerganizer), a column heater (set at 55 ℃), a Diode Array Detector (DAD), and a column as specified in the methods below. The flow from the column was split to the MS spectrometer. The MS detector is equipped with an electrospray ionization source. Mass spectra were acquired by scanning from 100 to 1000 in 0.18 seconds using a dwell time of 0.02 seconds. The capillary needle voltage was 3.5kV and the source temperature was maintained at 140 ℃. Nitrogen was used as the atomizing gas. Data acquisition was performed using a Waters-Micromass MassLynx-Openlynx data System.

General procedure B

HPLC measurements were performed using an Alliance HT 2790(Waters) system comprising a quaternary pump equipped with a degasser, an autosampler, a column oven (set at 40 ℃ C., unless otherwise specified), DAD and columns as specified in the methods below. The flow from the column was split to the MS spectrometer. The MS detector is equipped with an electrospray ionization source. Mass spectra were acquired by scanning from 100 to 1000 in 1 second using a dwell time of 0.1 second. The capillary needle voltage was 3kV and the source temperature was maintained at 140 ℃. Nitrogen was used as the atomizing gas. Data acquisition was performed using a Waters-MicromassMassLynx-Openlynx data system.

LCMS method 1

In addition to general procedure a: reverse phase UPLC was performed on a bridged ethylsiloxane/hybrid silica (BEH) C18 column (1.7 microns, 2.1x50 millimeters; Waters Acquity) using a flow rate of 0.8 ml/min. Two mobile phases (25mmol concentration of NH) were used₄OAc at H₂O/CH₃In CN 95/5; mobile phase B: CH (CH)₃CN) was operated from 95% a and 5% B to 5% a and 95% B at 1.3 min and gradient conditions were maintained for 0.3 min. An injection volume of 0.5 microliters was used. The cone voltage for the positive ionization mode is 10V and the negative ionization mode is 20V.

LCMS method 2

In addition to general procedure a: reverse phase UPLC was performed on a BEH C18 column (1.7 micron, 2.1x50 mm; waters acquity) using a flow rate of 0.8 ml/min. Two mobile phases were used (mobile phase A: 0.1% formic acid in H)₂O/MeOH 95/5; mobile phase B: MeOH) was run from 95% a and 5% B to 5% a and 95% B at 1.3 min and held at a gradient condition of 0.2 min. An injection volume of 0.5 microliters was used. The cone voltage for the positive ionization mode is 10V and the negative ionization mode is 20V.

LCMS method 3

In addition to general procedure B: reverse phase HPLC was performed on an Atlantis C18 column (3.5 microns, 4.6X100 mm) using a flow rate of 1.6 ml/min. Two mobile phases (mobile phase A: 70% MeOH + 30% H) were used₂O; mobile phase B: 0.1% formic acid in H₂O/MeOH 95/5) was operated from 100% B to 5% B + 95% a at 9 minutes and these conditions were maintained over a gradient of 3 minutes. An injection volume of 10 microliters was used. The cone voltage for the positive ionization mode is 10V and the negative ionization mode is 20V.

LCMS method 4

In addition to general procedure a: reverse phase UPLC (Ultra Performance Liquid Chromatography) was performed on a BEH C18 column (1.7 micron, 2.1x50 mm; waters acquity) using a flow rate of 0.8 ml/min. Two mobile phases (25mmol concentration of NH) were used₄OAc/CH₃CN 95/5; mobile phase B: CH (CH)₃CN) was operated from 95% a and 5% B to 5% a and 95% B at 1.3 min and gradient conditions were maintained for 0.3 min. An injection volume of 0.5 microliters was used. The cone voltage for the positive ionization mode is 30V and the negative ionization mode is 30V.

LCMS method 5

In addition to general procedure B: the column heater was set at 60 ℃. Reverse phase HPLC was performed on an Xterra MS C18 column (3.5 μm, 4.6 × 100 mm) using a flow rate of 1.6 ml/min. Three mobile phases were used (mobile phase A: 95% NH 25mmol concentration₄OAc+5％CH₃CN; mobile phase B: CH (CH)₃CN; mobile phase C: MeOH) was run from 100% a to 50% B and 50% C at 6.5 min to 100% B at 0.5 min and these conditions were held for 1 min and the gradient conditions were re-equilibrated with 100% a for 1.5 min. An injection volume of 10 microliters was used. The cone voltage for the positive ionization mode is 10V and the negative ionization mode is 20V.

Melting Point

The melting points (m.p.) of several compounds were determined using DSC823e (Mettler-Toledo). Melting points were measured using a temperature gradient of 30 ℃/min. The maximum temperature was 400 ℃. The listed values are the highest values.

The results of the analytical measurements are shown in Table 2.

Table 2: residence time in minutes (R)_t)，[M+H]⁺Peak (protonated molecule), LCMS method and m.p. (melting point in ° c). (n.d. means no measurement)

¹H NMR

For several compounds CHLOROFORM-d (deuterated CHLOROFORM, CDCl) was used₃) Or DMSO-d₆(deuterated DMSO, dimethyl-d 6 sulfoxide) as solvent, recorded on a Bruker DPX-360 or Bruker DPX-400 spectrometer equipped with a standard pulse sequence and operating at 360MHz and 400MHz respectively¹H NMR spectrum. Chemical shifts (δ) are reported in parts per million relative to Tetramethylsilane (TMS) as an internal standard.

Co.No.1：(360MHz，CDCl₃)δppm 2.31(s，3H)，3.84(s，3H)，3.92(s，3H)，6.89(s，1H)，6.92-7.01(m，4H)，7.02-7.09(m，1H)，7.09-7.15(m，1H)，7.19(d，J＝8.3Hz，1H)，7.24(d，J＝8.3Hz，1H)，7.39-7.48(m，2H)，7.49-7.56(m，1H)，7.64(s，1H)，8.38(s，1H).

Co.No.2：(360MHz，DMSO-d₆)δppm 2.14(s，3H)，2.64(s，3H)，3.75(s，3H)，3.84(s，3H)，6.92-7.00(m，2H)，7.01(s，1H)，7.09-7.16(m，3H)，7.19(d，J＝8.5Hz，1H)，7.23-7.31(m，3H)，7.52(t，J＝8.4Hz，1H)，7.63(d，J＝1.1Hz，1H)，8.33(s，1H).

Co.No.3：(360MHz，DMSO-d₆)δppm 0.91(t，J＝7.3Hz，3H)，1.27(sxt，J＝7.3Hz，2H)，1.92(quin，J＝7.3Hz，2H)，2.34(s，3H)，3.79(s，3H)，4.44(t，J＝6.9Hz，2H)，6.91-7.01(m，2H)，7.10(d，J＝7.3Hz，1H)，7.16(d，J＝2.2Hz，1H)，7.28(d，J＝8.4Hz，1H)，7.37(d，J＝8.4Hz，1H)，7.65(s，1H)，8.38(s，1H)，8.54(br.s.，1H)，9.28(d，J＝1.5Hz，1H)，14.84(br.s.，1H).

Co.No.4：(360MHz，CDCl₃)δppm 2.61(s，3H)，2.65(s，3H)，3.88(s，3H)，6.89(s，1H)，6.99-7.06(m，2H)，7.10-7.16(m，2H)，7.20(d，J＝8.1Hz，1H)，7.46(t，J＝8.4Hz，1H)，7.58(d，J＝8.8Hz，1H)，7.72(dd，J＝8.8，2.6Hz，1H)，7.83(s，1H)，8.64(d，J＝2.6Hz，1H).

Co.No.5：(360MHz，DMSO-d₆)δppm 2.39(s，3H)2.64(s，3H)3.80(s，3H)3.84(s，3H)6.95-7.04(m，2H)7.09-7.19(m，3H)7.22-7.35(m，3H)7.43(d，J＝8.78Hz，1H)7.52(t，J＝8.42Hz，1H)9.05(br.s.，1H).

Co.No.6：(360MHz，CDCl₃)δppm 2.53(s，3H)3.95(s，3H)5.00(q，J＝8.42Hz，2H)6.83(s，1H)6.92(d，J＝1.83Hz，1H)6.99(dd，J＝8.42，1.83Hz，1H)7.06(t，J＝7.68Hz，1H)7.14(d，J＝6.95Hz，1H)7.18(d，J＝8.42Hz，1H)7.30(s，1H)7.67(d，J＝8.42Hz，1H)8.01(s，1H).

Co.No.7：(360MHz，DMSO-d₆)δppm 2.63(s，3H)3.84(s，3H)3.87(s，3H)6.98(t，J＝7.68Hz，1H)7.07(d，J＝7.32Hz，1H)7.09-7.17(m，3H)7.22-7.32(m，3H)7.48-7.58(m，2H)7.79(s，1H)7.93-8.03(m，1H)8.41(s，1H).

Co.No.8：(360MHz，DMSO-d₆)δppm 2.02(br.s.，2H)2.45(s，3H)4.06(s，2H)6.94-7.02(m，1H)7.08(d，J＝7.32Hz，1H)7.31(s，1H)7.35(m，J＝8.78Hz，2H)7.43(d，J＝6.22Hz，1H)7.44-7.50(m，2H)7.53(m，J＝8.42Hz，2H)7.84-7.92(m，2H)8.41(s，1H).

Co.No.9：(360MHz，CDCl₃)δppm 2.30(s，3H)，3.82(s，3H)，4.23(s，3H)，6.76(s，1H)，6.88(s，1H)，6.90-6.96(m，2H)，7.01(t，J＝7.7Hz，1H)，7.11(d，J＝7.3Hz，1H)，7.14-7.22(m，2H)，7.63(s，1H)，7.88(s，1H).

Co.No.10：(360MHz，DMSO-d₆)δppm 0.92(t，J＝7.3Hz，3H)，1.32(sxt，J＝7.3Hz，2H)，1.85(quin，J＝7.3Hz，2H)，2.34(s，3H)，2.63(s，3H)，3.79(s，3H)，4.37(t，J＝7.3Hz，2H)，6.87-6.99(m，2H)，7.09(d，J＝7.2Hz，1H)，7.14(d，J＝2.2Hz，1H)，7.26(d，J＝8.3Hz，1H)，7.37(d，J＝8.6Hz，1H)，7.64(s，1H)，8.48(br.s.，1H)，9.28(d，J＝1.5Hz，1H)，15.01(br.s.，1H).

Co.No.11：(360MHz，DMSO-d₆)δppm 0.91(t，J＝7.32Hz，3H)1.28(sxt，J＝7.32Hz，2H)1.94(quin，J＝7.32Hz，2H)2.35(d，J＝0.73Hz，3H)3.82(s，3H)4.50(t，J＝7.32Hz，2H)7.08-7.15(m，2H)7.27(d，J＝2.20Hz，1H)7.47(d，J＝8.78Hz，1H)7.68(t，J＝1.10Hz，1H)7.91(d，J＝1.10Hz，1H)8.65(s，1H)8.89(br.s.，1H)9.32(d，J＝1.46Hz，1H)14.99(br.s.，1H).

Co.No.12：(400MHz，DMSO-d₆)δppm 0.91(t，J＝7.27Hz，3H)1.26(sxt，J＝7.27Hz，2H)1.92(quin，J＝7.27Hz，2H)2.36(s，3H)3.83(s，3H)4.01(q，J＝5.65Hz，2H)4.44(t，J＝6.86Hz，2H)7.16(dd，J＝8.68，2.22Hz，1H)7.23(d，J＝1.21Hz，1H)7.24(d，J＝2.02Hz，1H)7.35(s，1H)7.40(d，J＝8.48Hz，1H)7.63(t，J＝1.21Hz，1H)8.42(br.s.，2H)8.45(s，1H)8.59(br.s.，1H)9.29(d，J＝1.61Hz，1H)15.04(br.s.，1H).

Co.No.13：(360MHz，CDCl₃)δppm 0.54-0.63(m，2H)0.77-0.85(m，2H)1.47-1.61(m，1H)2.54(s，3H)3.87(s，3H)4.44(d，J＝7.32Hz，2H)6.94-7.09(m，3H)7.14-7.23(m，2H)7.30-7.39(m，2H)7.90(br.s.，1H)8.22(s，1H)8.40(br.s.，1H).

Co.No.14：(360MHz，CDCl₃)δppm 1.21(d，J＝6.9Hz，6H)，1.48(t，J＝6.9Hz，3H)，2.00(s，3H)，2.30(s，3H)，2.42(s，3H)，3.23-3.43(m，1H)，3.80(s，3H)，4.11(q，J＝6.9Hz，2H)，6.80(s，1H)，6.87(s，1H)，6.90-6.96(m，3H)，6.99-7.06(m，1H)，7.11(s，1H)，7.13-7.21(m，3H)，7.63(d，J＝1.3Hz，1H).

Co.No.15：(360MHz，CDCl₃)δppm 2.30(s，3H)，3.81(s，3H)，5.57(s，2H)，6.83(s，1H)，6.88(s，1H)，6.90-6.97(m，2H)，7.00-7.13(m，4H)，7.14-7.19(m，2H)，7.23-7.28(m，2H)，7.63(s，1H)，7.87(s，1H).

Co.No.16：(360MHz，CDCl₃)δppm 2.31(s，3H)，2.34(t，J＝1.8Hz，3H)，3.83(s，3H)，6.89(d，J＝5.1Hz，2H)，6.92-6.99(m，2H)，7.01-7.10(m，2H)，7.14(d，J＝6.9Hz，1H)，7.19(d，J＝8.3Hz，1H)，7.25(d，J＝6.9Hz，1H)，7.64(d，J＝1.1Hz，1H)，7.81(td，J＝8.8，5.8Hz，1H)，8.37(d，J＝2.7Hz，1H).

Co.No.17：(360MHz，CDCl₃)δppm 2.49(s，3H)3.87(s，3H)4.23(s，3H)6.81(s，1H)6.94-6.99(m，2H)7.03(t，J＝7.68Hz，1H)7.12(d，J＝6.95Hz，1H)7.20(d，J＝8.05Hz，1H)7.58(d，J＝9.15Hz，1H)7.88(s，1H)8.48(s，1H).

Co.No.19：(360MHz，CDCl₃)δppm 2.50(s，3H)，3.90(s，3H)，3.93(s，3H)，6.93-7.09(m，5H)，7.13(d，J＝7.3Hz，1H)，7.24(d，J＝7.3Hz，1H)，7.42-7.48(m，2H)，7.50-7.53(m，1H)，7.62(d，J＝8.3Hz，1H)，8.38(s，1H)，8.50(s，1H).

Co.No.20：(360MHz，CDCl₃)δppm 1.21(d，J＝6.6Hz，6H)，1.48(t，J＝7.0Hz，3H)，2.00(s，3H)，2.42(s，3H)，2.49(s，3H)，3.33(spt，J＝6.8，6.6Hz，1H)，3.86(s，3H)，4.11(q，J＝7.0Hz，2H)，6.79(s，1H)，6.92-6.96(m，2H)，6.97-7.06(m，2H)，7.11(s，1H)，7.15-7.22(m，2H)，7.56(d，J＝8.8Hz，1H)，8.46(s，1H).

Co.No.22：(360MHz，CDCl₃)δppm 2.10-2.32(m，4H)，2.50(s，3H)，2.63(s，2H)，4.42(t，J＝6.7Hz，2H)，6.79(s，1H)，6.96-7.05(m，1H)，7.05-7.23(m，4H)，7.67(t，J＝8.4Hz，1H)，8.42(d，J＝2.6Hz，1H).

Co.No.23：(360MHz，CDCl₃)δppm 2.34(s，3H)，2.51(s，3H)，6.96(s，1H)，7.01-7.26(m，5H)，7.31(d，J＝8.4Hz，1H)，7.70(t，J＝8.8Hz，1H)，7.80(td，J＝8.8，5.8Hz，1H)，8.38(d，J＝2.6Hz，1H)，8.44(d，J＝2.6Hz，1H).

Co.No.24：(360MHz，CDCl₃)δppm 2.52(s，3H)4.23(s，3H)6.78(br.s.，1H)7.00(t，J＝7.87Hz，1H)7.07-7.13(m，2H)7.17(d，J＝8.42Hz，1H)7.32(m，J＝8.42Hz，2H)7.56(m，J＝8.78Hz，2H)7.86(s，1H).

Co.No.25：(360MHz，DMSO-d₆)δppm 0.42-0.51(m，2H)0.54-0.63(m，2H)1.33-1.52(m，1H)2.50(s，3H)4.30(d，J＝7.32Hz，2H)6.91-6.99(m，1H)7.04(d，J＝7.32Hz，1H)7.24(d，J＝8.05Hz，1H)7.34(m，J＝8.78Hz，2H)7.42(s，1H)7.54(m，J＝8.78Hz，2H)8.41(s，1H).

Co.No.26：(360MHz，DMSO-d₆)δppm 1.80-1.98(m，2H)2.47(s，3H)2.48-2.56(m，2H)2.61-2.76(m，2H)5.16(quin，J＝8.42Hz，1H)6.95(t，J＝8.42，7.32Hz，1H)7.02(d，J＝7.32Hz，1H)7.20(d，J＝8.05Hz，1H)7.29-7.39(m，3H)7.54(m，2H)8.44(s，1H).

Co.No.27：(360MHz，DMSO-d₆)δppm 2.47(s，3H)3.25(s，3H)3.84(t，J＝5.12Hz，2H)4.59(t，J＝5.12Hz，2H)6.95(t，J＝8.05，7.32Hz，1H)7.02(d，J＝7.32Hz，1H)7.22(d，J＝8.05Hz，1H)7.29-7.39(m，3H)7.53(m，2H)8.33(s，1H).

Co.No.28：(360MHz，CDCl₃)δppm 2.14-2.38(m，4H)2.52(s，3H)3.62(td，J＝11.25，3.11Hz，2H)4.10-4.27(m，2H)4.57-4.72(m，1H)6.84(s，1H)7.00(t，J＝7.68Hz，1H)7.07-7.13(m，2H)7.18(d，J＝8.05Hz，1H)7.34(m，2H)7.56(m，2H)7.94(s，1H).

Co.No.29：(360MHz，DMSO-d₆)δppm 2.45(s，3H)，2.64(s，3H)，3.84(s，3H)，6.93-7.00(m，1H)，7.07(d，J＝6.9Hz，1H)，7.10-7.15(m，1H)，7.23-7.29(m，3H)，7.31(s，1H)，7.35(d，J＝8.8Hz，2H)，7.46-7.58(m，3H)，8.39(s，1H).

Co.No.30：(360MHz，CDCl₃)δppm 1.21(d，J＝6.9Hz，6H)，1.47(t，J＝6.9Hz，3H)，2.00(s，3H)，2.41(s，3H)，2.52(s，3H)，3.33(spt，J＝6.9Hz，1H)，4.11(q，J＝6.9Hz，2H)，6.79(s，1H)，6.94(br.s.，1H)，6.99-7.05(m，1H)，7.09(s，1H)，7.11(s，1H)，7.16(d，J＝8.1Hz，2H)，7.32(m，J＝8.4Hz，2H)，7.54(m，J＝8.4Hz，2H).

Co.No.31：(360MHz，DMSO-d₆)δppm 2.46(s，3H)，2.61(s，3H)，6.94-7.00(m，1H)，7.08(d，J＝7.0Hz，1H)，7.26(d，J＝8.3Hz，1H)，7.30-7.39(m，3H)，7.46(t，J＝8.8Hz，2H)，7.53(d，J＝8.8Hz，2H)，7.70-7.81(m，2H).

Co.No.32：(360MHz，DMSO-d₆)δppm 2.47(s，3H)7.21(dd，J＝6.59，1.46Hz，1H)7.28-7.38(m，3H)7.42(d，J＝8.78Hz，2H)7.52-7.64(m，4H)7.96-8.08(m，2H)8.97(s，1H).

Co.No.33：(360MHz，CDCl₃)δppm 2.34(t，J＝1.8Hz，3H)，2.53(s，3H)，6.91(s，1H)，7.00-7.09(m，2H)，7.11(s，1H)，7.13(d，J＝7.3Hz，1H)，7.23(d，J＝8.1Hz，1H)，7.35(m，J＝8.8Hz，2H)，7.58(m，J＝8.4Hz，2H)，7.82(td，J＝8.8，5.8Hz，1H)，8.36(d，J＝2.6Hz，1H).

Co.No.34：(360MHz，CDCl₃)δppm 2.53(s，3H)6.08(s，2H)6.93(d，J＝8.42Hz，1H)6.96(s，1H)7.00-7.07(m，1H)7.09-7.13(m，2H)7.20(d，J＝8.05Hz，1H)7.32(dd，J＝8.42，2.20Hz，1H)7.35(m，2H)7.42(d，J＝2.20Hz，1H)7.57(m，2H)8.25(s，1H).

Co.No.35：(360MHz，DMSO-d₆)δppm 2.47(s，3H)2.60(s，3H)3.85(s，3H)7.09(d，J＝6.22Hz，1H)7.14-7.20(m，1H)7.27-7.33(m，2H)7.37(s，1H)7.54(t，J＝8.23Hz，1H)7.59(m，2H)7.66(d，J＝6.22Hz，1H)8.22(m，2H)9.43(s，1H).

Co.No.36：(360MHz，DMSO-d₆)δppm 2.10(s，3H)3.25(s，3H)3.76(s，3H)3.85(t，J＝5.31Hz，2H)4.60(t，J＝5.31Hz，2H)6.90-6.99(m，2H)7.06(d，J＝1.83Hz，1H)7.11(d，J＝6.95Hz，1H)7.20(d，J＝8.42Hz，1H)7.24(d，J＝8.05Hz，1H)8.28(s，1H)8.34(s，1H).

Co.No.37：(360MHz，DMSO-d₆)δppm 0.40-0.53(m，2H)0.53-0.65(m，2H)1.31-1.53(m，1H)2.10(s，3H)3.76(s，3H)4.30(d，J＝7.32Hz，2H)6.92-7.00(m，2H)7.06(d，J＝1.83Hz，1H)7.11(d，J＝7.32Hz，1H)7.20(d，J＝8.42Hz，1H)7.24(d，J＝8.05Hz，1H)8.27(s，1H)8.40(s，1H).

Co.No.38：(360MHz，DMSO-d₆)δppm 0.42-0.50(m，2H)0.54-0.63(m，2H)1.29-1.51(m，1H)2.46(s，3H)3.87(s，3H)4.30(d，J＝7.32Hz，2H)6.92-7.01(m，2H)7.06-7.11(m，2H)7.21(s，1H)7.23(d，J＝8.05Hz，1H)7.49(d，J＝8.42Hz，1H)8.39(s，1H).

Co.No.39：(400MHz，DMSO-d₆)δppm 2.10(s，3H)2.64(s，3H)3.76(s，3H)3.84(s，3H)6.94-7.03(m，2H)7.07(d，J＝2.02Hz，1H)7.10-7.18(m，2H)7.21(d，J＝8.07Hz，1H)7.24-7.31(m，3H)7.52(t，J＝8.28Hz，1H)8.25(s，1H).

Co.No.40：(360MHz，DMSO-d₆)δppm 2.45(s，3H)，2.62(s，3H)，3.86(s，3H)，6.98-7.05(m，2H)，7.09(d，J＝2.2Hz，1H)，7.14(d，J＝6.9Hz，1H)，7.19(s，1H)，7.27(d，J＝8.1Hz，1H)，7.41-7.53(m，3H)，7.71-7.82(m，2H)，8.39(s，1H).

Co.No.42：(360MHz，CDCl₃)δppm 2.52(s，3H)2.61(s，3H)3.90(s，3H)4.02(s，3H)6.93(d，J＝6.22Hz，1H)7.04-7.16(m，3H)7.32(s，1H)7.38(dd，J＝8.42，1.83Hz，1H)7.49(t，J＝8.23Hz，1H)7.63-7.69(m，2H)7.79(d，J＝6.22Hz，1H)7.98(d，J＝1.83Hz，1H).

Co.No.43：(360MHz，DMSO-d₆)δppm 0.38-0.50(m，2H)0.54-0.65(m，2H)1.30-1.50(m，1H)2.53(s，3H)4.28(br.s.，2H)7.02(t，J＝7.68Hz，1H)7.11(d，J＝6.95Hz，1H)7.42(d，J＝8.05Hz，1H)7.72-7.87(m，2H)7.87-7.98(m，1H)8.43(s，1H)8.47(s，1H)9.22(br.s.，1H).

Co.No.44：(360MHz，CDCl₃)δppm 2.57(s，3H)5.01(q，J＝8.42Hz，2H)6.79(s，1H)7.01-7.11(m，2H)7.23(dd，J＝7.68，1.83Hz，1H)7.43(s，1H)7.57(d，J＝8.42Hz，1H)7.71(dd，J＝8.42，2.56Hz，1H)8.03(s，1H)8.61(d，J＝2.56Hz，1H).

Co.No.46：(360MHz，CDCl₃)δppm 2.61(s，3H)，2.62(s，3H)，6.85(s，1H)，6.97-7.07(m，1H)，7.12(d，J＝6.9Hz，1H)，7.19(d，J＝8.1Hz，1H)，7.22-7.32(m，2H)，7.52-7.61(m，3H)，7.71(dd，J＝8.8，2.9Hz，1H)，7.83(s，1H)，8.64(d，J＝2.6Hz，1H).

Co.No.48：(360MHz，CDCl₃)δppm 1.41(d，J＝6.95Hz，6H)3.15(spt，J＝6.95Hz，1H)5.00(q，J＝8.29Hz，2H)6.82(s，1H)7.05(t，J＝7.68Hz，1H)7.10(d，J＝6.95Hz，1H)7.13(s，1H)7.17(d，J＝8.05Hz，1H)7.34(m，J＝8.42Hz，2H)7.59(m，J＝8.42Hz，2H)8.01(s，1H).

Co.No.49：(360MHz，CDCl₃)δppm 2.71(s，3H)3.90(s，3H)4.22(s，3H)6.81(s，1H)6.88-6.96(m，2H)7.02(t，J＝7.68Hz，1H)7.13(d，J＝7.68Hz，1H)7.17(d，J＝8.42Hz，1H)7.49(d，J＝8.42Hz，1H)7.86(s，1H)7.90(s，1H).

Co.No.50：(360MHz，CDCl₃)δppm 2.34(t，J＝2.0Hz，3H)，2.72(s，3H)，3.92(s，3H)，6.90-6.95(m，2H)，6.97(dd，J＝8.2，2.0Hz，1H)，7.01-7.08(m，2H)，7.16(d，J＝6.9Hz，1H)，7.24(dd，J＝8.4，0.7Hz，1H)，7.51(d，J＝8.1Hz，1H)，7.82(td，J＝8.5，6.0Hz，1H)，7.92(s，1H)，8.36(d，J＝2.6Hz，1H).

Co.No.51：(360MHz，DMSO-d₆)δppm 2.43(s，3H)2.47(s，3H)3.91(s，3H)5.46(q，J＝9.03Hz，2H)6.85(s，1H)7.15(dd，J＝8.42，1.83Hz，1H)7.22(d，J＝1.83Hz，1H)7.30(s，1H)7.61(d，J＝8.42Hz，1H)8.47(s，1H)9.11(s，1H).

Co.No.52：(360MHz，CDCl₃)δppm 2.31(d，J＝0.73Hz，3H)5.00(q，J＝8.42Hz，2H)6.84(s，1H)6.93(q，J＝1.46Hz，1H)7.03-7.09(m，2H)7.13(dd，J＝7.32，0.73Hz，1H)7.20(dd，J＝12.62，2.38Hz，1H)7.23-7.30(m，2H)7.66(t，J＝1.46Hz，1H)8.03(s，1H).

Co.No.53：(360MHz，DMSO-d₆)δppm 2.64(s，3H)3.84(s，3H)3.86(s，3H)6.34(d，J＝1.83Hz，1H)6.98(t，J＝7.86Hz，1H)7.06-7.16(m，2H)7.23-7.29(m，3H)7.35-7.43(m，4H)7.45(d，J＝1.83Hz，1H)7.52(t，J＝8.23Hz，1H).

Co.No.54：(360MHz，CDCl₃)δppm 2.30(d，J＝0.73Hz，3H)4.07(s，3H)5.01(q，J＝8.42Hz，2H)6.60(d，J＝8.05Hz，1H)6.89(t，J＝1.10Hz，1H)7.12(dd，J＝8.60，7.50Hz，1H)7.22-7.29(m，1H)7.44(d，J＝8.42Hz，1H)7.56(s，1H)7.64(d，J＝1.10Hz，1H)8.03(s，1H)8.05(d，J＝7.32Hz，1H).

Co.No.55：(360MHz，CDCl₃d)δppm 2.31(s，3H)5.06(q，J＝8.05Hz，2H)6.58(s，1H)6.94(s，1H)7.10(dd，J＝8.60，2.38Hz，1H)7.19(dd，J＝12.26，2.38Hz，1H)7.30(t，J＝8.60Hz，1H)7.68(s，1H)8.25(s，1H)8.33(s，1H)8.83(s，1H).

Co.No.56：(360MHz，CDCl₃)δppm 1.32(d，J＝6.95Hz，6H)2.51(s，3H)2.96-3.13(m，J＝13.79，6.89，6.89，6.89，6.89Hz，1H)3.93(s，3H)4.98(q，J＝8.29Hz，2H)6.87(s，1H)7.02-7.14(m，3H)7.75(d，J＝8.42Hz，1H)8.15(s，1H)8.58(s，1H).

Co.No.57：(360MHz，DMSO-d₆)δppm 1.27(d，J＝6.95Hz，6H)2.16(s，3H)3.05(spt，J＝6.95Hz，1H)4.06(s，3H)5.47(q，J＝9.15Hz，2H)7.13(s，1H)7.16(d，J＝8.42Hz，1H)7.74(d，J＝8.42Hz，1H)7.77(d，J＝1.10Hz，1H)8.26(s，1H)8.57(s，1H)9.82(s，1H).

Pharmacology of

A) Screening of Compounds of the invention for Gamma-secretase-modulating Activity

A1) Method 1

Screening was performed using SKNBE cells grown in Dulbecco's modified Eagle's Medium/Nutrient mixture F-12(DMEM/NUT-mix F-12) (HAM) and harboring APP 695-wild type, supplied by a Dulbecco's modified Eagle's Medium/Nutrient (Dulbecco's modified Eagle's Medium/Nutrient) containing 5% serum/Fe supplemented with 1% non-essential amino acids. Cells were grown to near confluence.

The methods used in Citron et al (1997) Nature Medicine 3: 67 were screened. Briefly, the day before the addition of the compound, cells were plated at about 10⁵Individual cells/ml were plated onto 96-well plates. The compounds were added to cells in Ultraculture (Lonza, BE12-725F) supplemented with 1% glutamine (Invitrogen, 25030-024) over 18 hours. Media were tested for a β 42 and total a β (a β total) by two sandwich elisas (sandwich elisas). Compounds were tested for toxicity by WST-1 cell proliferation agent (Roche, 1644807) according to the manufacturer's protocol.

To quantify the amount of A.beta.42 in the cell supernatant, a commercially available Enzyme-Linked Immunosorbent Assay (ELISA) kit (Innotest) was usedβ-Amyloid_(1-42)Innogenetics n.v., Ghent, Belgium). The a β 42ELISA was performed essentially according to the manufacturer's protocol. Briefly, a standard (a dilution of synthetic a β 1-42) was prepared in polypropylene Eppendorf and the final concentration was 8000 drops to 3.9 pg/ml (1/2 dilution step). Samples, standards and blanks (100 microliters) were added to anti- Α β 42-coated plates (capture antibody selectively recognising the C-terminus of the antigen) supplied with the kit. The plate was incubated at 25 ℃ for 3 hours to allow formation of antibody-amyloid complexes. Following this incubation and subsequent washing steps, a selective anti- Α β -antibody conjugate (biotinylated 3D6) was added and incubated for at least 1 hour to allow formation of antibody-amyloid-antibody complexes. After incubation and appropriate washing steps, streptavidin-Peroxidase-Conjugate (streptavidin-Peroxidase-Conjugate) was added, and after 30 minutes 3, 3 ', 5, 5' -Tetramethylbenzidine (TMB)/peroxyl benzidine (TMB) was addedA mixture of substances resulting in the conversion of the substrate into a colored product. The reaction was stopped by adding sulfuric acid (0.9N) and the color intensity was measured by luminometer equipped with ELISA-reader with 450nm filter.

To quantify the total a β (a β total) amount in the cell supernatant, samples and standards were added to 6E 10-coated plates. The plate was incubated at 4 ℃ overnight to allow formation of antibody-amyloid complexes. Following this incubation and subsequent washing steps, a selective anti- Α β -antibody conjugate (biotinylated 4G8) was added and incubated for at least 1 hour to allow formation of antibody-amyloid-antibody complexes. After incubation and appropriate washing steps, streptavidin-Peroxidase-Conjugate (streptavidin-Peroxidase-Conjugate) was added, and after 30 minutes the Quanta Blu fluoroperoxidase substrate (Pierce corp., Rockford, Il) was added according to the manufacturer's instructions.

To obtain the values reported in table 3a, sigmoidal dose-response curves were analyzed by computerized curve-fitting, and the percentage of inhibition was plotted against compound concentration. IC determination Using the 4-parameter equation in XLfit (model 205)₅₀. The top and bottom of the curve are fixed at 100 and 0, respectively, and the slope is fixed at 1. IC (integrated circuit)₅₀Represents the concentration of the compound required to inhibit the biological effect by 50% (here, it is the concentration at which the level of a β peptide is reduced by 50%).

IC₅₀The values are shown in Table 3 a:

A2) method 2

Screening was performed using NBE2 cells grown in a Dulbecco's Modified Eagle's Medium/Nutrient mixture F-12(DMEM/NUT-mix F-12) (HAM) and harboring the APP 695-wild type, supplied by Invitrogen (cat No.10371-029) containing 5% serum/Fe supplemented with 1% non-essential amino acids, 1-glutamine 2mM, Hepes15mM, penicillin 50U/ml (units/ml) and erythromycin 50. mu.g/ml. Cells were grown to near confluence.

The methods used in Citron et al (1997) Nature Medicine 3: 67 were screened for modifications from the assay described in. Briefly, cells were treated at 10 in the presence of different test concentrations of test compound⁴Each cell/well was placed in 384-well plates supplemented with 1% glutamine (Invitrogen, 25030-024), 1% non-essential amino acids (NEAA), penicillin 50U/ml and erythromycin 50. mu.g/ml. The cell/compound mixture was incubated at 37 ℃ and 5% CO₂Incubate overnight. The next day the media was tested for a β 42 and total a β (a β total) by two sandwich immuno-assays.

Aphalisa technology (Perkin Elmer) was used to quantify the A β 42 and total A β (A β total) concentrations in cell supernatants. Aphalisa is a sandwich assay using biotinylated antibodies attached to streptavidin-coated donor beads and antibodies conjugated to acceptor beads. In the presence of antigen, the beads are in close proximity. Excitation of the donor beads results in the release of singlet oxygen molecules that initiate an energy transfer cascade in the acceptor beads resulting in luminescence. To quantify the amount of a β 42 in the cell supernatant, a C-terminal specific monoclonal antibody to a β 42 (JRF/cA β 42/26) was coupled to acceptor beads and reacted with donor beads using biotinylated antibodies (JRF/a β N/25) that gave specificity to the N-terminus of a β. To quantify the total a β (Α β total) amount in the cell supernatant, a monoclonal antibody specific for Α β N-terminus (JRF/Α β N/25) was coupled to acceptor beads and reacted with donor beads using a biotinylated antibody specific for Α β middomain (biotinylated 4G 8).

To obtain the values reported in table 3b, data were calculated as a percentage of the maximum amount of amyloid β 42 measured in the absence of test compound. Sigmoidal dose-response curves were analyzed using non-linear regression analysis, plotted as percent of control versus log concentration of compound. Determination of IC Using 4-parameter equation₅₀。

IC₅₀Values are shown in table 3b ('n.d.' indicates no assay):

B) demonstration of in vivo efficacy

The a β 42-lowering agents of the invention may be used to treat AD in mammals such as humans or demonstrate efficacy in animal models such as, but not limited to, mice, rats or guinea pigs. The mammal may not be diagnosed with AD, or may not have a genetic predisposition to AD, but may have a transgene that overproduces and eventually deposits a β in a similar manner as seen in humans suffering from AD.

The a β 42 lowering agent can be administered in any standard form using any standard method. For example, but not limited to, the a β 42 lowering agent may be in the form of a liquid, tablet or capsule that is administered orally or by injection. The a β 42 lowering agent can be administered at any dose sufficient to significantly reduce a β 42 level in blood, plasma, serum, cerebrospinal fluid (CSF) or brain.

To determine whether acute administration of an a β 42-lowering agent will lower a β 42 levels in vivo, non-transgenic rodents, such as mice or rats, are used. Alternatively, two to three month old Tg2576 mice expressing APP695 containing the "Swedish" variant or a transgenic mouse model developed by dr.fred Van Leuven (k.u.leuven, Belgium) and colleagues can be used with clinical mutations that are neuron-specific for expression of human amyloid precursor protein [ V717I ] (Moechars et al, 1999 j.biol.chem.274, 6483). Young transgenic mice had high levels of a β in the brain, but no detectable a β deposition. At about 6-8 months of age, the transgenic mice begin to develop a spontaneous, progressive accumulation of β -amyloid (a β) in the brain, eventually leading to amyloid plaques in the subbrain (subculum), hippocampus, and cortex. Animals treated with a β 42 lowering agents are examined and the soluble a β 42 and total a β levels in the brain are quantified by standard techniques, such as using ELISA, compared to those animals that are not treated or treated with vehicle. The duration of treatment varies from a few hours to several days and is adjusted according to the outcome of the a β 42 reduction (once the time course of onset is determined).

A typical protocol for measuring a β 42 decline in vivo is presented, but it is just one of many that can be used to optimize the changes in detectable a β levels. For example, compounds with reduced a β 42 are formulated as 20% Captisol in water(a sulfobutyl ether of beta-cyclodextrin) or 20% hydroxypropyl beta cyclodextrin. The a β 42-lowering agent is administered as a single oral dose or by any acceptable route of administration to an overnight fasted animal. After 4 hours, animals were sacrificed and analyzed for a β 42 levels.

Blood was collected into EDTA-treated collection tubes by decapitation and exsanguination. Blood was centrifuged at 1900g for 10 minutes (min) at 4 ℃ and plasma was recovered and flash frozen for subsequent analysis. The brain was removed from the skull and hindbrain. The cerebellum is removed and the left and right hemispheres are separated. The left hemisphere was stored at-18 ℃ for quantitative analysis of the levels of test compounds. The right hemisphere was washed with Phosphate Buffered Saline (PBS) buffer and immediately frozen on dry ice and stored at-80 ℃ until homogenized for biochemical testing.

Mouse brains were resuspended in 10 volumes of 0.4% DEA (diethylamine)/50 mM NaCl pH 10 (for non-transgenic animals) or 0.1% 3- [ (3-cholamidopropyl (cholamidopropyl)) dimethylamino (aminomonopy) ] -1-propanesulfonate (CHAPS) (for transgenic animals) in Tris Buffered Saline (TBS) containing protease inhibitors per gram of tissue (Roche-11873580001 or 04693159001), e.g., 0.158g brain plus 1.58ml of 0.4% DEA. All samples were sonicated on ice at 20% power output (pulse mode) for 30 seconds. The homogenate was centrifuged at 221.300Xg for 50 minutes. The resulting high-speed supernatant is then transferred to a new tube and optionally further purified before the next step. A portion of the supernatant was neutralized with 10% 0.5M Tris-HCl and used to quantify total A β (A β total).

The resulting supernatant was purified using a Water Oasis HLB reversed phase column (Waters corp., Milford, MA) to remove non-specific immunoreactive material from brain lysates prior to subsequent a β detection. Using a vacuum manifold, the entire solution was passed through the column at a rate of about 1ml per minute, so the vacuum pressure was adjusted accordingly throughout the procedure. The column was pre-stabilized with 100% MeOH (preconditioned) and then with 1ml of H₂And (4) balancing the oxygen. The non-neutralized brain lysis lysate was packed in the column. The filled sample was then washed a first time with 1ml of 5% MeOH, twice repeatedly and a second time with 1ml of 30% MeOH. Finally, use a catalyst containing 2% NH₄OH 90% MeOH eluted a β from the column into a 100x30mm glass tube. The eluate is then transferred to a 1.5ml tube and concentrated in a speed-vacuum concentrator at an elevated temperature of 70 ℃ for about 1.5-2 hours. The concentrated A.beta.was then resuspended in UltraCULTURE Universal Serum-Free Medium (General Purpose Serum Free-Serum Medium) (Cambrex Corp., Walkersville, Md.) and added with a protease inhibitor according to the manufacturer's recommendations.

To quantify the amount of A.beta.in the soluble fraction of brain homogenate, a commercially available Enzyme-Linked Immunosorbent Assay (ELISA) kit (e.g., Innotest) was usedβ-Amyloid_(1-42)Innogenetics n.v., Ghent, Belgium). A β 42ELISA was performed using only the plates provided with the kit. Briefly, standards (dilutions of synthetic A β 1-42) were prepared in Ultraculture in 1.5ml Eppendorf tubes at final concentrations ranging from 25000 to 1.5 pg/ml. Samples, standards and blanks (60 microliters) were added to anti- Α β 42-coated plates (capture antibody selectively recognising the C-terminus of the antigen). The plates were incubated overnight at 4 □ to allow formation of antibody-amyloid complexes. After this incubation and subsequent washing step, the optional anti-Abeta-antibody conjugate (raw) is addedBiotinylated detection antibody such as biotinylated 4G8) (Covance research products, Dedham, MA) and incubated for at least 1 hour to allow formation of antibody-amyloid-antibody complexes. After incubation and appropriate washing steps, streptavidin-Peroxidase-Conjugate (streptavidin-Peroxidase-Conjugate) was added, and after 50 minutes the Quanta Blu fluorescent Peroxidase substrate was added according to the manufacturer's instructions (Pierce corp., Rockford, Il). Dynamic readings were performed every 5 minutes for 30 minutes (excitation 320 nm/emission 420 nm). To quantify the total a β (Α β total) in the soluble fraction of brain homogenate, samples and standards were added to JRF/rA β/2-coated plates. The plate was incubated at 4 ℃ overnight to allow formation of antibody-amyloid complexes. ELISA was then performed for a β 42 detection.

In this mode, at least a 20% a β 42 reduction would be advantageous compared to untreated animals.

The results are shown in Table 4:

C) effect of the Gamma-secretase-Complex on Notch-processing Activity

Notch-free (Notch) cell assay

The Notch transmembrane domain releases the Notch intracellular C-terminal domain (NICD) by cleavage with gamma secretase. Notch is a signaling protein that plays an important role in the development process and therefore is preferably a compound that does not exhibit an effect on the Notch-processing activity of the γ -secretase complex.

To monitor the effect of compounds on NICD production, recombinant Notch substrates were prepared (N99). A Notch substrate containing a mouse Notch fragment (V1711-E1809), N-terminal methionine and C-terminal FLAG sequence (DYDDDK) was expressed in E.coli and purified on a column containing an anti-FLAG M2 affinity matrix.

A typical nick-free (Notch) cell assay consists of 0.3-0.5. mu.M nick (Notch) substrate, abundantly prepared gamma-secretase and 1. mu.M test compound (compound 45 of the invention). The control comprises a gamma-secretase inhibitor (GSI), such as (2S) -N- [2- (3, 5-difluorophenyl) acetyl]-L-alanyl-2-phenyl-glycine 1, 1-dimethylethyl ester (DAPT) or (2S) -2-hydroxy-3-methyl-N- [ (1S) -1-methyl-2-oxo-2- [ [ (1S) -2, 3, 4, 5-tetrahydro-3-methyl-2-oxo-1H-3-benzazepine-1-yl]Amino group]Ethyl radical]-butyramide (Semagacestat), and DMSO, with a final concentration of DMSO of 1%. The recombinant Notch substrate was pretreated with 17 μ MDTT (1, 4-dithiothreitol) and 0.02% SDS (sodium dodecyl sulfate) and heated at 65 ℃ for 10 min. The substrate, gamma-secretase and compound/DMSO mixture was incubated at 37 ℃ for 6 to 22 hours. The 6 hours of incubation was sufficient to produce the maximum amount of NICD and the cleaved product remained stable for an additional 16 hours. The reaction product was processed for SDS PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) and Western blotting. The blots were sequentially assayed with anti-Flag M2 antibody and LI-COR Infrared secondary antibody probe and used with the Orditosyl Infrared Imaging System (LI-COR)Biosciences).

In the Notch-free (Notch) cell assay, none of the test compounds (compound 45 of the invention) inhibited cleavage of C99 by gamma-secretase, whereas production of NICD was prevented by control GSI (DAPT or semagastatt). Thus, it was demonstrated that compound 45 showed no effect on the nick (Notch) -processing activity of the γ -secretase-complex (resulting in NICD).

Composition examples

The "active ingredient (a.i.)" used in these examples relates to a compound of formula (I), including any stereochemically isomeric form thereof, a pharmaceutically acceptable salt thereof or a solvate thereof; in particular any of the exemplified compounds.

Typical examples of the formulations of the invention are as follows:

1. tablet formulation

2. Suspension liquid

Aqueous suspensions for oral administration are prepared so that each ml contains 1 to 5mg of the active ingredient, 50mg of sodium carboxymethylcellulose, 1mg of sodium benzoate, 500mg of sorbitol and added water to adjust to 1 ml.

3. Injection solution

Parenteral compositions are prepared by stirring 1.5% (w/v) of the active ingredient in 0.9% NaCl solution or in 10% by volume propylene glycol in water.

4. Ointment

In this example, the active ingredient may be replaced by the same amount of any of the compounds according to the invention, in particular by the same amount of any of the exemplified compounds.

Reasonable variations are not to be regarded as a departure from the scope of the invention. The invention thus described may obviously be modified in many ways by a person skilled in the art.

Claims

1. A compound of the formula (I),

or a stereoisomeric form thereof, wherein

X¹is CH or N;

X²is CR⁵Or N;

X³is CR⁶Or N;

provided that X is¹，X²And X³No more than two of which are N;

A¹is CR⁷Or N; wherein R is⁷Is hydrogen, halo or C_1-4An alkoxy group;

R⁸Is hydrogen or C_1-4An alkyl group;

R⁹is hydrogen or C_1-4An alkyl group;

R¹⁰is hydrogen or C_1-4An alkyl group;

R¹¹is hydrogen or C_1-4An alkyl group;

R¹²is C_1-4An alkyl group;

G¹is O or S;

G²is CH or N;

or a pharmaceutically acceptable addition salt or solvate thereof.

2. The compound according to claim 1, or a stereoisomeric form thereof, wherein

R¹Is optionally substituted by one or more radicals each independently selected from halo, C_1-4Alkoxy radical, C_3-7Cycloalkyl and phenyl substituted C_1-6An alkyl group;

C_3-7cycloalkyl, tetrahydropyranyl, 1, 3-benzodioxolyl or phenyl;

wherein each phenyl group is independently selected from one or more of halo, C_1-4Alkyl and C_1-4Substituent substitution of alkoxy;

R²is hydrogen, cyano, or optionally substituted by one or more NH groups₂C substituted by substituents_1-4An alkyl group;

X²is CR⁵Or N; in particular X²Is CR⁵；

R⁵Is hydrogen, halo, cyano, or optionally substituted by one or more NH groups₂C substituted by substituents_1-4An alkyl group;

X³is CH or N;

A²is CH or N and A³And A⁴Is CH;

Het¹is a 5-membered aromatic heterocycle having the formula (a-1), (a-2), (a-3) or (a-4);

R¹⁰is C_1-4An alkyl group;

R¹¹is hydrogen;

R⁸is hydrogen;

R¹²is C_1-4An alkyl group;

or a pharmaceutically acceptable addition salt or solvate thereof.

3. The compound according to claim 1, or a stereoisomeric form thereof, wherein

R¹Is covered by a C_1-4Phenyl substituted with an alkoxy substituent; or R¹Is C substituted by one or more halo substituents_1-6An alkyl group;

R²is hydrogen;

X¹，X²and X³Is CH;

A¹is CR⁷(ii) a Wherein R is⁷Is C_1-4An alkoxy group; a. the²，A³And A⁴Is CH;

Het¹has the formula (a-1) or (a-2);

G¹is O; g²Is CH;

R⁸is C_1-4An alkyl group;

R¹⁰is C_1-4An alkyl group;

R⁹is hydrogen;

or a pharmaceutically acceptable addition salt or solvate thereof.

4. The compound according to claim 1, or a stereoisomeric form thereof, wherein

R¹Is selected from one or more of C independently_1-4Alkyl and C_1-4Phenyl substituted with a substituent of alkoxy;

or a pharmaceutically acceptable addition salt or solvate thereof.

5. The compound according to claim 1, or a stereoisomeric form thereof, wherein

R¹Is C optionally substituted by one or more halo substituents_1-6An alkyl group;

or a pharmaceutically acceptable addition salt or solvate thereof.

6. The compound according to claim 1, wherein said compound is selected from the group consisting of N- [ 3-methoxy-4- (2-methyl-5-)Azolyl) phenyl]-2- (2, 2, 2-trifluoroethyl) -2H-indazol-7-amine, and N- [ 3-methoxy-4- (4-methyl-1H-imidazol-1-yl) phenyl]-2- (3-methoxyphenyl) -3-methyl-2H-indazol-7-amine,

including any stereochemically isomeric form thereof,

and pharmaceutically acceptable addition salts and solvates thereof.

7. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and, as active ingredient, a therapeutically effective amount of a compound as defined in any one of claims 1 to 6.

8. A compound as defined in any one of claims 1 to 6 for use as a medicament.

9. A compound as defined in any one of claims 1 to 6 for use in the treatment or prevention of a disease or condition selected from alzheimer's disease, traumatic brain injury, mild cognitive impairment, senility, dementia associated with lewy bodies, amyloid cerebrovascular disease, multi-infarct dementia, down's syndrome, dementia associated with parkinson's disease and dementia associated with beta-amyloid.

10. A compound according to claim 9, wherein the disease is alzheimer's disease.