SI21099A - N-(5-(((5-alkyl-2-oxazolyl)methyl)thio)-2-thiazolyl)carboxamide inhibitors of cyclin dependent kinases - Google Patents

Abstract

The present invention describes compounds of the formula: and enantiomers, diasteromers, solvates, and pharmaceutically acceptable salts thereof. The formula compounds are protein kinase inhibitors and are useful in the treatment of proliferative diseases, for example, cancer, inflammation and arthritis. They may also be useful in the treatment of Alzheimer's disease, chemotherapy-induced alopecia, and cardiovascular disease.

Description

The present invention relates to compounds of formula I

(I) and their enantiomers, diastereomers, solvates and pharmaceutically acceptable salts, wherein R is alkyl;

R ^{1 is} hydrogen or alkyl;

X NR ² or CHNR ² R ³ ;

R ² and R ³ are each independently hydrogen, alkyl, substituted alkyl, cycloalkyl or substituted cycloalkyl; and n is 0, 1, 2 or 3.

The compounds of formula I are particularly useful as potent protein kinase inhibitors and are useful in the treatment of proliferative diseases such as cancer, inflammation and arthritis. They may also be useful in the treatment of Alzheimer's disease, chemotherapy-induced alopecia and cardiovascular disease.

The present invention describes compounds of formula I, pharmaceutical compositions that include such compounds, and methods for using such compounds.

The definitions of the various terms used to describe the compounds of the present invention are given below. These definitions apply to terms as used throughout the description (unless otherwise limited in certain cases), either individually or as part of a larger group.

The term "alkyl" or "alk" refers to a derived monovalent alkane (hydrocarbon) radical containing from 1 to 12, preferably 1 to 6, and more preferably 1 to 4, carbon atoms, unless otherwise defined. The alkyl group is an optionally substituted straight, branched or cyclic saturated hydrocarbon group. When substituted, the alkyl groups may be substituted by up to four substituent groups, R ⁴ being as defined at any available point of attachment. Where it is stated that the alkyl group is substituted by an alkyl group, this is used interchangeably with a "branched alkyl group". Examples of unsubstituted such groups include methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl, isobutyl, pentyl, hexyl, isohexyl, heptyl, 4,4-dimethylpentyl, octyl, 2,2,4 trimethylpentyl, nonyl, decyl, undecyl, dodecyl etc. Examples of substituents may include, but are not limited to, one or more of the following groups: halo (such as F, Cl, Br or J), haloalkyl (such as CCI3 or CF3), alkoxy, alkylthio, hydroxy, carboxy, alkylcarbonyl, alkyloxycarbonyl, alkylcarbonyloxy , amino, carbamoyl, urea, amidinyl or thiol.

Cycloalkyl is a type of alkyl containing from 3 to 15 carbon atoms without alternating or resonating double bonds between carbon atoms. May contain 1 to 4 rings. Examples of unsubstituted such groups include cyclopropyl, cyciobutyl, cyclopentyl, cyclohexyl, etc. Examples of substituents include one or more of the following groups: halogen, alkyl, alkoxy, alkyl hydroxy, amino, nitro, cyano, thiol and / or alkylthio.

The terms "alkoxy" or "alkylthio" as used herein mean an alkyl group, as described above, linked to an oxygen bond (-O-) or. sulfur bond (-S-).

The term "alkyloxycarbonyl" as used herein means an alkoxy group bonded through a carbonyl group. The alkoxycarbonyl radical is represented by the formula: -C (O) OR ⁵ , wherein the R ⁵ group is a straight or branched C 1-6 alkyl group.

The term "alkylcarbonyl" refers to an alkyl group attached to a carbonyl group.

The term " alkylcarbonyloxy ", as used herein, means an oxygen-bonded alkylcarbonyl group.

As used herein, the phrase " compounds of the invention " means together the compounds of formula I and their pharmaceutically acceptable salts and solvates, including hydrates. The processes of salt formation, solvation and hydrate formation are known in the art. The invention also includes mixtures of stereoisomers of the compounds of the invention. Stereoisomers include, but are not limited to, enantiomers, diastereomers and racemates, where the compound has one or more chiral centers. All stereoisomers of the compounds of this invention, whether in admixture or in pure or substantially pure form, are considered. The definition of the compounds of the invention encompasses all possible stereoisomers and mixtures thereof. In particular, it comprises racemic forms and isolated activity isomers of optical isomers. Racemic forms can be distinguished by physical methods, such as. fractional crystallization, separation or crystallization of diastereomeric derivatives or separation by chiral column chromatography. Individual optical isomers can be obtained from racemates by conventional procedures such as e.g. salt formation with optically active acid followed by crystallization. All configuration isomers of the compounds of the invention, whether in admixture or in pure or substantially pure form, are considered. The definition of the compounds of the invention particularly encompasses both c / s (Z) and trans (E) alkene isomers, as well as cis and trans isomers of cycloalkyl or heterocycloalkyl rings.

In addition, the present invention also includes salts of compounds of formula I, which are pharmaceutically unsuitable but useful in other aspects, e.g. for the separation or purification of Formula I compounds

The compounds of the invention are defined herein by their chemical structures and / or chemical names. Where a compound is referred to by both chemical structure and chemical name and where the chemical structure and chemical name are in conflict, the chemical structure is decisive for the identity of the compound.

The phrase "pharmaceutically acceptable salt (s)" as used herein includes, but is not limited to, salts of the acid or base groups which may be present in the compounds of the invention. Examples of such pharmaceutically acceptable salts include, but are not limited to, hydrochloride, hydrobromomide, dihydrochloride, sulfate, trifluoroacetate, tartrate, fumarate, succinate, maleate, citrate, methanesulfonate, bromate and iodate salts, and mixtures thereof. Also included are salts formed with other organic and inorganic acids such as hydroxymethane sulfonic acid, acetic acid, benzenesulfonic acid, toluenesulfonic acid and many others, e.g. nitrates, phosphates, borates, benzoates, ascorbates, salicylates and the like. In addition, the pharmaceutically acceptable salts of the invention of formula I may be formed with alkali metals such as sodium, potassium and lithium; alkaline earth metals such as calcium and magnesium; organic bases such as dicyclohexylamine, tributylamine and pyridines and the like; and amino acids such as arginine, lysine and the like.

The salts of the compounds of the invention include solvates, racemates, and all their stereoisomeric forms, including enantiomers and diastereomers (e.g., D-tartrate and L-tartrate salts).

As used herein, the term "solvate" means a compound of the invention or a salt thereof, which further includes a stoichiometric or non-stoichiometric amount of one or more soluble molecules bound by non-covalent intermolecular forces. Preferred solvents are volatile, non-toxic and / or acceptable for human traceability. When the solvent is water, the solvate is called "hydrate".

The compounds of formula I can be prepared by following the procedures described in WO 99/65884 and WO 99/24416, which are incorporated herein by reference.

Alternatively, the generic method shown in Scheme A below, which shows the synthesis of a broader genus of compounds of Formula XIV, can be used to synthesize compounds of Formula I. The starting compounds are commercially available but can be prepared by methods known in the art. In Scheme A, the following terms appear:

R ⁷ , R ⁸ and R ¹⁰ are independently hydrogen or alkyl;

R is alkyl, aryl or heteroaryl;

R ⁹ is hydrogen, alkyl, aryl or heteroaryl;

R ¹ and R ¹¹ are independently hydrogen, alkyl, aryl, heteroaryl, halogen, hydroxy or alkoxy;

R ¹² is hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, CONR ¹³ R ¹⁴ , COR ¹⁵ or COOR ¹⁶ ;

R ¹³ , R ¹⁴ , R ¹⁵ and R ¹⁶ are independently hydrogen, alkyl or aryl;

r is an integer between 0 and 5;

s is an integer between 0 and 5;

L is the appropriate departure group, e.g. halogen or sulfonate (R ⁶ SO ₂ O, CF ₃ SO ₂ O, etc., wherein R ^{6 is} alkyl, cycloalkyl or aryl);

M is hydrogen, Li, Na, K, Cs or a quaternary ammonium ion, e.g. (R ⁶ ) ₄ N or quaternary ammonium ions containing cyclic alkentetramines, such as e.g. hexamethylenetetramine;

Q is hydroxy, halogen or acyloxy (R ⁶ COO, R ⁶ OCOO, etc.);

Y is O, S, NH, N-alkyl, N-aryl or N-acyl;

Z is hydrogen, alkyl, aryl, O-alkyl, O-aryl, S-alkyl, S-aryl, NH2, N-alkyl, N-aryl or N-acyl; and

P is a nitrogen protecting group (Boc, Cbz, R ₃ Si, etc.). When a function group is named "protected", it means that the group is in a modified form that eliminates unwanted side reactions in a protected location. Appropriate protecting groups for the compounds included in these processes will be described in the description, taking into account the level of knowledge in the art and referring to standard textbooks such as e.g. Greene, TW Protective Groups and Organic Synthesis, 3rd Edition (1999), incorporated herein by reference.

The processes generally involve reacting α-halo ketones II (commercially available or synthesized by well-known methods) with azide to obtain α-azido ketones III. Reduction of lil with a reducing reagent gives α-amino ketones IV.

Alternatively and more preferably, α-amino ketones IV are prepared by reacting α-halo ketones II with cyclic alkylenetetramine such as hexamethylenetetramine and the like, followed by hydrolysis of the resulting, new quaternary ammonium salt III '. This reaction gives excellent yields of the desired intermediate IV above 90%.

Then, by reacting α-amino ketones IV with α-halo acyl halide V in the presence of a base, or alternatively by coupling α-amino ketones IV with α-halo acid, the corresponding amides VI are obtained. Then, closing ring VI with a dehydration reagent yields 2-oxaolylalkyl halides VII. When a conventional dehydration reagent such as trihalophosphorus oxide such as POCI3 is used, isolating the product is difficult due to the formation of large amounts of hydrochloric and phosphoric acid. In the process of the present invention, Burgess reagent is preferably used which gives excellent yields and allows easy, safe isolation of the product from water.

By further treatment with 2-oxazolylalkyl halides VII with sulfur-containing reagent Vlil or

New key intermediates, 2-oxazolylalkyl sulfides IX, are obtained from Vyl. Combining IX with 5-halo-aminothiazole X yields 5- (2-oxazolylalkylthio) -2-aminothiazoles XI. Coupling XI with an azacycloalkanoic acid derivative XII yields thiazolyl amides XIII that can be deprotected (in the case where P is a protecting group, e.g., Boc) to give 5- (2oxazolylalkylthio) -2-azacycloalkanoylaminothiazoles XIV.

Scheme A

As shown in Scheme A, the processes for the preparation of 5- (2-oxazolylalkylthio) -2azacycloalkanoylaminothiazoles and analogs include the following transformations:

Step (a) involves reacting α-substituted ketone II, such as e.g. α-halo ketone, with azide in a suitable solvent or solvent mixtures to give α-azido ketone III; or a more desirable (a ') reaction of α-substituted ketone II, such as α-halo ketone with cyclic alkylenetetramine, such as e.g. hexamethylenetetramine in a suitable solvent or solvent mixtures to obtain a new quaternary ammonium salt lil '.

α-halo ketone includes α-halo aliphatic and α-halo aromatic ketones. Preferred α-halo ketones are α-halo pinacolones, with α-bromo pinacolone being the most desirable. Sulfonate, e.g. RSO2O- (where R is alkyl, aryl or heteroaryl), CF3SO2O- and the like, can substitute for halogen in the α-position. Azides include both metal azides and quaternary ammonium azides. Metal azides are more desirable and sodium azide is most desirable. Suitable solvent (s) include solvents such as hydrocarbons, ethers, amides, e.g. dimethylformamide, ketones, etc. or mixtures thereof with ketones such as acetone, which is advantageous for both reactions (a) and (a ') Step (b) comprises reacting the α-azido ketone III obtained in step (a) with a reducing reagent in a suitable solvent, or solvent mixtures to obtain α-amino ketone IV, or more preferably (b ') reacting the quaternary ammonium salt of lil' obtained in step (a ') with an acid in a suitable solvent, or solvent mixtures to obtain α-amino ketone IV.

The reducing reagent in reaction (b) includes hydrogen in the presence of a transition metal catalyst such as palladium, trialkyl or triarylphosphines such as triphenylphosphine. Hydrogen in the presence of a transition metal catalyst is preferred, and even more preferred is hydrogen and palladium over activated carbon. A suitable solvent in reaction (b) includes solvents such as hydrocarbons, ethers, alcohols and the like, or mixtures thereof, preferably with an alcohol such as methanol. Alternatively, the reduction reaction may be carried out in the presence of an acidic medium, such as e.g. hydrochloric acid in ethanol to give an α-amino ketonic acid salt that can be isolated as an acid salt or free amino forms.

The acid in reaction (b ') includes, but is not limited to, protic acids such as HCl, HBr, HJ,

H2SO4, H3PO4, etc., preferably HCl. Suitable solvents in reaction (b ') include solvents such as hydrocarbons, ethers, alcohols and the like, or mixtures thereof, with alcohol, preferably ethanol. The α-amino ketone product can be isolated as salt or free base forms.

Step (C) involves reacting (acylating) the α-amino ketone IV or its acid salts obtained in step (b) or (b ') with an α-substituted acyl derivative V, such as e.g. a-halo acyl halide, in the presence of a base and in a suitable solvent or solvent mixtures to give the amide

VI.

α-halo acyl halide V includes α-alkyl or aryl substituted or unsubstituted α-halo acyl halide, the latter having the advantage. The most preferred α-halo acyl halide is acloroacetyl chloride. The base used in this reaction includes, but is not limited to, aromatic or aliphatic organic amines, the latter being preferred. The most preferred base is triethylamine. Suitable solvents include aprotic solvents such as hydrocarbons, halogenated hydrocarbons, ethers, esters and the like, or mixtures thereof, with halogenated hydrocarbons such as dichloromethane. Alternatively, the reaction may be carried out using substituted acid instead of the α-substituted acyl derivative, and then a coupling reagent such as a water-soluble diimide such as carbodiimide, haloformate, thionyl halide, etc. may be used. In any reaction, the sulfonate, e.g. RSO ₂ O- (where R is alkyl, aryl or heteroaryl), CF3SO2-O- and the like are substituted for halogen at the α-position of the α-halo acyl halide or α-halo acid reactants shown.

Step (d) concerns the reaction of amide VI obtained in step (c) with a dehydration reagent in a suitable solvent or solvent mixtures to give cyclized 2-oxazolylalkyl derivative VII, such as e.g. 2-Oxazolylalkyl halide.

The preferred reaction is carried out using (methoxycarbonylsulfamoyl) -triethylammonium hydroxide (Burgess reagent) as a dehydration reagent. Suitable solvents include hydrocarbons, halogenated hydrocarbons, ethers and the like or mixtures of volatiles. The use of Burgess reagent in tetra id rofu ran u is most desirable. Suitable dehydration reagents include, but are not limited to, bases, acids, acid anhydrides, and the like, e.g. concentrated sulfuric acid, polyphosphoric acid, etc. Although less convenient, the dehydration reagent may be e.g. trihalophosphorus oxide, such as tribromophosphorus oxide or trichlorophosphorus oxide, alone or with a solvent such as e.g. toluene.

Step (e) is directed to the reaction of 2-oxazolylalkyl derivative VII obtained in step (d) with sulfur-containing reagent Vlil or Vlil 'in a suitable solvent or solvent mixtures to give 2-oxazolylalkyl sulfide IX, a new key intermediate.

The sulfur-containing reagent includes N-substituted or unsubstituted thioureas, thio acids or salts such as thioacetic acid or its salts, xanthic acids or salts such as potassium salt of ethylxanthic acid. Unsubstituted thiourea is preferred. Suitable solvents include hydrocarbons, halogenated hydrocarbons, ethers, esters, amides, alcohols and the like, or mixtures thereof with an alcohol such as methanol or preferably ethanol.

Step (f) concerns the reaction of 2-oxazolylalkyl sulfide IX obtained in step (e) with 5halo-2-aminothiazole X in the presence of a base and in a suitable solvent or solvent mixtures to give 5- (2-oxazolylalkylthio) -2-aminothiazole XI.

5-halo-2-aminothiazole includes 4-N-substituted or unsubstituted 5-halo-2aminothiazoles, preferably 5-bromo-2-aminothiazoles. Suitable bases include, but are not limited to, metal hydroxide, metal alkoxides, metal carbonates, and aqueous amines such as ammonium hydroxide. Sodium hydroxide is preferred. Suitable solvents include solvents such as hydrocarbons, halogenated hydrocarbons, ethers, esters, amides, alcohols and the like, or mixtures thereof, with halogenated hydrocarbons such as dichloromethane.

Step (g) involves reacting 5- (2-oxazolylalkylthio) -2-aminothiazole XI obtained in step (f) with an azacycloalkanoic acid derivative XII in the presence of a coupling reagent in a suitable solvent or solvent mixtures to give thiazolyl amide XIII.

The azacycloalkanoic acid derivative includes N-protected derivatives, for example N-protected isonipecotinic acid or N-protected nipectic acid. Preferred nitrogen protecting groups are Boc, Cbz, silicon derivatives and the like, with Boc being preferred.

-1010

The coupling reagent includes, but is not limited to, water-soluble carbodiimides, haloformates and the like, preferably carbodiimides such as alkylcarbodiimides. Suitable solvents include solvents such as hydrocarbons, halogenated hydrocarbons, ethers, esters, amides, etc. and or mixtures thereof, with halogenated hydrocarbons such as dichloromethane.

Step (h) is directed to reacting the thiazolyl amide XII obtained in step (g) with a deprotection reagent in a suitable solvent or solvent mixture to obtain the desired 5 (2-oxazolylalkylthio) -2-azacycloalkanoylaminothiazole XIV (wherein R ¹¹ is hydrogen ).

The choice of deprotection reagent depends on the nature of the protecting group (P). For the Boc protecting group, the preferred deprotection reagent is an acid such as e.g. hydrochloric acid or trifluoroacetic acid and suitable solvents for such a deprotection reaction include solvents such as hydrocarbons, halogenated hydrocarbons, ethers, esters, amides, etc. or mixtures thereof, with halogenated hydrocarbons such as dichloromethane.

A more detailed synthesis of the compounds of formula I is shown in Schemes 1-5 below. The starting compounds are commercially available or may be prepared by one skilled in the art by methods known in the art. Schemes 1-5 below use the following terms:

L is a suitable leaving group such as halogen or sulfonate (e.g., Br, Cl, J, R ⁶ SC> 20, CF ₃ SO ₂ O, wherein R ^{6 is} alkyl, cycloalkyl, heteroaryl or aryl);

M is hydrogen, Li, Na, K, Cs or a quaternary ammonium ion, e.g. (R ⁶ ) ₄ N or quaternary ammonium ions which include cyclic alkentetramines such as hexamethylenetetramine;

Q is hydroxy, halogen or acyloxy (R ⁶ COO, R ^S OCOO, etc.);

Y is O, S, NH, N-alkyl, N-aryl or N-acyl; and

Z is hydrogen, alkyl, aryl, O-alkyl, O-aryl, S-alkyl, S-aryl, NH ₂ , N-alkyl, N-aryl or N-acyl. Scheme 1 shows the synthesis of compounds of formula 11.

-1111

Scheme 1: Synthesis of Compounds of Formula 11

First, step (a) involves reacting a suitable cc-substituted ketone 2, such as an achalo ketone, with the azide in a suitable solvent or solvent mixtures to give α-azido ketone 3; or more preferably (a ¹ ) reacting ketone 2 with cyclic alkylentetramine, such as hexamethylenetetramine in a suitable solvent or solvent mixtures, to give quaternary ammonium salt 3 '.

Suitable α-halo ketones 2 include α-halo aliphatic and α-halo aromatic ketones. Preferred α-halo ketones are α-halo pinakoions, with α-bromo pinacolone being the most preferred. Sulfonate, e.g. R ⁶ SO ₂ O (wherein, as defined above, is R ⁶ alkyl, cycloalkyl, heteroaryl or aryl), CF ₃ SO ₂ O and the like may be substituted for halogen (as L group) at the α-position. Azides include both metal azides and quaternary

-1212 ammonium azides. Preference is given to metal azides, sodium azide being the most preferred.

Suitable solvents include hydrocarbons, ethers, esters, amides such as dimethylformamide, ketones, etc. or mixtures thereof, wherein ketones such as acetone are preferred for both reactions (a) and (a ').

Step (b) involves reacting the α-azido ketone 3 obtained in step (a) with a reducing reagent in a suitable solvent or solvent mixtures to obtain α-amino ketone 4, or more preferably (b ¹ ) reacting the quaternary ammonium salt 3 ' obtained in step (a ') with an acid in a suitable solvent or solvent mixtures to give the α-amino ketone 4.

The reducing reagent in reaction (b) includes hydrogen in the presence of a transition metal catalyst such as palladium, trealkyl- or trarylphosphines such as triphenylphosphine. Hydrogen in the presence of a transition metal catalyst is preferably preferred with hydrogen or palladium over activated carbon. Suitable solvents in reaction (b) include hydrocarbons, ethers, alcohols and the like, or mixtures thereof, with alcohols such as methanol being preferred. Alternatively, the reduction reaction may be carried out in the presence of an acidic medium such as hydrochloric acid in ethanol to give the acid salt of an amine ketone, which can be isolated as an acid salt or free amine forms.

Suitable acids for use in reaction (b ') include, but are not limited to, HCI, HBr, HJ, H ₂ SO ₄ , H3PO4, etc., preferably HCI. Suitable solvents in reaction (b ') include hydrocarbons, ethers, alcohols and the like, or mixtures thereof, whereby alcohols such as ethanol are the preferred α-amino ketone product which can be isolated as a salt or free base.

Step (c) comprises reacting (acylating) the α-amino ketone 4 or its acid salt obtained in step (b) or (b ') with an α-substituted acyl derivative 5 such as α-halo acyl halide (i.e. Q and L = halo), in the presence of a base and in a suitable solvent or solvent mixtures to give amide 6.

α-halo acyl halide 5 includes alkyl or aryl-α-halo acyl halides (substituted or unsubstituted), the latter being preferred. The most preferred α-halo acyl halide is α-1313 chloroacetyl chloride. The base used in the reaction includes, but is not limited to, aromatic and aliphatic organic amines, with the latter being preferred. The most preferred base is triethylamine. Suitable solvents include aprotic solvents such as hydrocarbons, halogenated hydrocarbons, ethers, esters and the like, or mixtures thereof with halogenated hydrocarbons such as dichloromethane. Alternatively, the reaction may be carried out using substituted acid (Q = OH) instead of an α-substituted acyl derivative, and then a coupling reagent such as water-soluble diimide (e.g. carbodiimide), haloformate, thionyl halide, etc. may be used. In any reaction, the sulfonate, e.g. R ^{6 is} SO ² O (where R ^{6 is} alkyl, cycloalkyl, aryl or heteroaryl), CF ₃ SO ₂ O substitutes halogen in the α-position (i.e. in group L) of compounds 5.

Step (d) involves reacting the amide 6 obtained in step (c) with a dehydration reagent in a suitable solvent or solvent mixtures to give a cyclized 2-oxazolylalkyl derivative 7, e.g. 2-Oxazolylalkyl halide (i.e. L is halo).

The reaction is preferably carried out using (methoxycarbonylsulfamoyl) -triethylammonium hydroxide (Burgess reagent) as a dehydration reagent. Suitable solvents include hydrocarbons, halogenated hydrocarbons, ethers and the like, or mixtures thereof. Most preferred is the use of a Burgess reagent in tetrahydrofuran. Suitable dehydration reagents include, but are not limited to, other bases, acids, acid anhydrides, and the like, such as concentrated sulfuric acid, polyphosphoric acid, etc. Dehydration trihalophosphorus oxide, such as tribromophosphorus oxide or trichlorophosphorus oxide, may be less advantageous alone or with a solvent such as toluene.

Step (e) involves reacting the 2-oxazolylalkyl derivative 7 obtained in step (d) with sulfur-containing reagent 8 or 8 'in a suitable solvent or solvent mixtures to give 2-oxazolylalkyl sulfide 9.

The sulfur-containing reagent includes N-substituted or unsubstituted thioureas, thio acids or salts such as thioacetic acid or its salts, xanthic acids or salts such as potassium salt of ethylxanthic acid. Unsubstituted thiourea is preferred. Suitable solvents include hydrocarbons, halogenated hydrocarbons, ethers, esters, amides, alcohols

-1414 and the like or mixtures thereof, with preference being given to alcohols such as methanol or ethanol.

Step (f) shows the reaction of 2-oxazholylalkyl sulfide 9 obtained in step (e) with 2-aminothiazole 10 (preferably L is halo) in the presence of a base and a suitable solvent or solvent mixtures to give 5- (2-oxazolylalkylthio) - 2-aminothiazole 11.

2-Aminothiazole 10 includes 4- / V-substituted or unsubstituted 5-halo-2-aminothiazoles, preferably 5-bromo-2-aminothiazoles. Suitable bases include, but are not limited to, metal hydroxides, metal alkoxides, metal carbonates, and aqueous amines such as ammonium hydroxide. Sodium hydroxide is preferred. Suitable solvents include hydrocarbons, halogenated hydrocarbons, ethers, esters, amides, alcohols and the like, or mixtures thereof, preferably having halogenated hydrocarbons such as dichloromethane.

Scheme 2 shows the general synthesis of compounds of formula I via the reaction of an amine 11 with a carboxylic acid of formula 12 in the presence of a coupling agent. Suitable coupling agents include, but are not limited to, water-soluble carbodiimides, haloformates and the like, preferably carbodiimides such as alkylcarbodiimides, e.g. combination of 1- (3dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and base.

Scheme 2

-1515

coupling agent

V

(I)

Scheme 3 below illustrates the synthesis of compounds of formula I wherein X is NR ² and R ² is H. First, the amine of formula 11 is reacted with an N-protected carboxylic acid of formula 13 in the presence of a coupling agent to give an N-protected compound of formula 14. Then compound 14 is deprotected to give compounds of formula I. Suitable coupling agents include, but are not limited to, water-soluble carbodiimides, haloformates and the like, preferably carbodiimides such as alkylcarbodiimides, e.g. 1- (3-dimethylaminopropyl) -3ethylcarbodiimide hydrochloride and base.

-1616

Scheme 3

In the above scheme, P is a nitrogen protecting group (e.g., Boc, Cbz, R ₃ Si, etc.). When a functional group is protected, it means that the group is in a modified form to prevent adverse side reactions in a protected location. Appropriate protecting groups for the compounds involved in these processes will be clear from the description, taking into account the prior art and reference to standard textbooks such as Greene, TW, Protective Groups and Organic Synthesis, 3rd Edition (1999), which is here included as a reference. Preferred

-1717 nitrogen protecting groups are Boc, Cbz, silicon derivatives, preferably Boc. Suitable solvents include hydrocarbons, halogenated hydrocarbons, ethers, esters, amides, and the like, or mixtures thereof, preferably halogenated hydrocarbons such as dichloromethane. The choice of deprotection reagent depends on the nature of the protecting group (P). For the Boc protecting group, the preferred deprotection reagent is an acid such as e.g. hydrochloric acid or trifluoroacetic acid and suitable solvents for such a deprotection reaction include solvents such as hydrocarbons, halogenated hydrocarbons, ethers, esters, amides, etc. or mixtures thereof, with halogenated hydrocarbons such as dichloromethane.

Scheme 4 below illustrates the synthesis of compounds of formula I, wherein X is NR ² and R ² is 2,3 dihydroxypropyl, where the compound of formula I is reacted, wherein X is NR ² and R ² is hydrogen with glyceraldehyde in the presence of a reducing agent, such as is sodium triacetoxyborohydride and an alcohol such as methanol.

-1818

Scheme 5 below illustrates the synthesis of compounds of formula I wherein X is NR ² and R ² is 2 hydroxyethyl by reacting a compound of formula I wherein X is NR ² and R ² is hydrogen with 2 (bromoethoxy) trialkylsilane of formula 15. to obtain intermediate 16, and deprotection intermediate 16 with an acid such as hydrogen fluoride.

Scheme 5

(I)

-1919

Preferred compounds of formula I are those wherein:

R is alkyl;

R ¹ is hydrogen;

X is NR ² or CHNR ² R ³ ; and

R ² and R ³ are independently hydrogen, alkyl, substituted alkyl or cycloalkyl); and n is 2.

The first group of more preferred compounds of the invention are those of formula Ia:

(1a) and their enantiomers, diastereomers, solvates and pharmaceutically acceptable salts, wherein R ^{2 is} hydrogen, alkyl, substituted alkyl or cycloalkyl.

Another group of more preferred compounds of the present invention are those of formula Ib:

and enantiomers, diastereomers, solvates and pharmaceutically acceptable salts thereof, wherein R ^{2 is} hydrogen, alkyl, substituted alkyl or cycloalkyl.

-2020

and their enantiomers, diastereomers, solvates and pharmaceutically acceptable salts, wherein R ² and R ^{3 are} each independently hydrogen, alkyl, substituted alkyl or cycloalkyl.

In another embodiment, the compounds of formula I include, but are not limited to, those compounds listed in Table 1 below and their enantiomers, diastereomers, solvates and pharmaceutically acceptable salts.

Table 1: Compounds of the Invention

Name Structure N - [5 - [[[5- (1,1-dimethylethyl) -2- oxazolyl] methyl] thio] -2-thiazolyl] -4- piperidinecarboxamide (HjChC _JH with ^H 1 J (±) - N - [5 - [[[5- (1,1-dimethylethyl) -2oxazolyl] methyl] thio] -2-thiazolyl] -3- piperidinecarboxamide (HAQ (±) -1- (2,3-dihydroxypropyl) -N- [5- [[[5- (1,1-dimethylethyl) -2- oxazolyl] methyl] thio] -2-thiazolyl] -4- piperidinecarboxamide N - [5 - [[[5- (1,1-dimethylethyl) -2- oxazolyl] methylethyl] -2-thiazole yl-1- (1- methylethyl) -4-piperidinecarboxamide (H ₃ C) ₃ C ν-ΎΓ'Λ '

-2121

Table 1: (cont.)

1-cyclopropyl- / V- [5 - [[[5- (1,1- d and meti I eti 1) -2-o xazole i I] m et i I] t io] -2 - thiazolyl] -4-piperidinecarboxamide (H ₃ Q ₃ C / ΧχήΧ N - [5 - [[[5- (1,1-dimethylethyl) -2- oxazolyl] methyl] thio] -2-thiazolyl] -1- (2- hydroxyethyl) -4- piperidinecarboxamide · (R) - / V- [5 - [[[5- (1,1-dimethylethyl) -2- oxazolyl] methyl] thio] -2-thiazolyl] -3- piperidinecarboxamide (h ₃ q ₃ c. ζΚ · χ ^! γ> · - (S) -A / - [5 - [[[5- (1,1 -d imethyleth) -2- oxazole and I] ethy I] thio] -2-azo I and I] -3- piperidinecarboxamide <h ₃ c) ₃ c: c / s-4-amino-N - [5 - [[[5- (1,1- dimethylethyl) -2-oxazolyl] methyl] thio] -2- thiazolylcyclohexylcarboxamide (H ₃ C) ₃ C frans-4-amino- / V- [5 - [[[5- (1,1- dimethylethyl) -2-oxazolyl] methyl] thio] -2- thiazolylcyclohexylcarboxamide (H ₃ C) ₃ C

Preferred salts of the above compounds are hydrochloride, hydrobromide, dihydrochloride, sulfate, trifluoroacetate, tartrate, fumarate, succinate, maleate, citrate, methanesulfonate, bromate and iodate salts or mixtures thereof.

The present invention also includes methods based on the pharmacological properties of the compounds of the invention. The compounds of the invention have pharmacological properties; in particular, the compounds of formula I are protein kinase inhibitors such as cyclin-dependent kinases (cdks), e.g. cdc2 (cdk1), cdk2, cdk3, cdk4, cdk5, cdk6, cdk7 and cdk8. Thus, the present invention also covers the use of the compounds of the invention for

-2222 treatment, prevention and / or management of cancer, inflammation or inflammatory disease, arthritis,

Alzheimer's disease and cardiovascular disease. The invention also encompasses, in a more specific embodiment, the use of the compounds of the invention for the treatment, prevention and / or management of proliferative diseases or their symptoms. The invention also encompasses the use of the compounds of the invention in the treatment or prevention of topical or systemic fungal infections.

More specifically, the compounds of formula I are useful in the treatment of various cancers, including (but not limited to) the following:

- carcinoma, including carcinoma of the bladder, breast, spinal cord, kidney, liver, lung, ovaries, pancreas, stomach, cervix, thyroids, prostate and skin;

- Hematopoietic tumors of the lymphatic line, including acute lymphatic leukemia, Cell lymphoma and Burkett lymphoma;

- hematopoietic tumors of the myeloid lineage, including acute and chronic myelogenous leukemia and promyelocytic leukemia;

- tumors of mesenchymal origin, including fibrosarcoma and rhabdomyosarcoma; and

- other tumors, including melanoma, semina, teratocarcinoma, osteosarcoma, neuroblastoma and glioma.

Without being restricted by any theory because of the key role of cdks in regulating cell proliferation in general, inhibitors could act as reversible cytostatic agents that are useful in treating any disease process that is reflected in abnormal cell proliferation, e.g. neurofibromatosis, atherosclerosis, pulmonary fibrosis, arthritis, psoriasis, glomerulonephritis, restenosis after angioplasty or vascular surgery, hypertrophic scar formation, inflammatory bowel disease, transplant rejection, angiogenesis and endotoxic shock.

The invention also encompasses the use of the compounds of the invention in the treatment of Alzheimer's disease, as shown by the recent discovery that cdk5 is involved in the phosphorylation of tau protein (J. Biochem, 117, 741-749 (1995)).

The invention also encompasses the use of the compounds of the invention as inhibitors of other protein kinases, e.g. protein kinase C, her2, rafl, MEK1, MAP kinase, EGF receptor, PDGF receptor, IGF

The -2323 receptor, PI3 kinase, wee1 kinase, Src, Abl, VEGF, and loc are thus effective in treating diseases associated with other protein kinases.

The invention also encompasses the use of the compounds of the invention for inducing or inhibiting apoptosis, a process of physiological cell death critical to normal development and homeostasis. Alterations in apoptotic pathways contribute to the pathogenesis of various human diseases. The compounds of formula I as modulators of apoptosis will be useful in the treatment of various human diseases by aberration in apoptosis, including cancer (especially, but not limited to, follicular lymphomas, carcinomas with p53 mutations, hormone-dependent breast tumors, prostate and ovarian, and precancerous lesions, is familial adenomatous polyposis), viral infections (including but not limited to, herpesvirus, poxvirus, Epstein-Barr virus, Sindbis virus and adenovirus), autoimmune diseases (including but not limited to, systemic lupus, erythematosus, immune-induced glomerulonephritis, rheumatoiditis, rheumatoid , psoriasis, inflammatory bowel disease and autoimmune diabetes mellitus), neurodegenerative diseases (including but not limited to, Alzheimer's disease, AIDS-related dementia, Parkinson's disease, amyotrophic lateral sclerosis, retinitis pigmentosis, spinal muscular atrophy and cerebral degeneration) , myelodysplastic syndromes, aplastic anemia, myocardial infarctions associated with ische myocardial injury, stroke and reperfusion impairment, arrhythmia, atherosclerosis, liver diseases caused by toxin or alcohol, haematological diseases (including but not limited to chronic anemia and aplastic anemia), degenerative diseases of the musculoskeletal system (including, but not limited to, osteoporosis and arthritis), aspirin-sensitive rhinosinusitis, cystic fibrosis, multiple sclerosis, kidney disease and cancer pain.

In another embodiment, the invention provides a method of inhibiting cdk in a cell. In particular, the present invention encompasses treating or preventing diseases associated with cdk modulation by administering one or more compounds of the invention to a mammal in need thereof.

The invention encompasses the treatment of mammals, especially humans.

In addition, the compounds of the invention may be used for the treatment of chemotherapy-induced alopecia, chemotherapy-induced thrombocytopenia, chemotherapy-induced leukopenia or mucocyitis. In chemotherapy treatment induced

-2424 alopecia administers the compounds of the invention preferably topically in the form of a medicament such as gel, solution, dispersion or cream.

The compounds of the present invention may be used in combination (before, during, after, including cyclic administration) with known anticancer treatments, such as radiation therapy or cytostatic or cytotoxic agents, including but not limited to, microtubule-stabilizing agents, microtubule-disrupting agents, alkylating agents, antimetabolites, epidophyllotoxins, antineoplastic enzymes, topoisomerase inhibitors, procarbazine, mitoxantrone, platinum coordination complexes, biological response modifiers, growth inhibitors, hormonal / antihormone therapeutical agents.

The classes of anticancer agents that can be used in combination with the compounds of the formula I of the present invention include, but are not limited to, the anthracycline drug family, vinca drug, mitomycin, bleomycin, cytotoxic nucleosides, taxanes, epothiolones, discodermolides, pteridine family of drugs, dienidine , aromatase inhibitors and podophyllotoxins. Special members of these classes include e.g. paclitaxel, docetaxel, 7-0methylthiomethylpaclitaxel (described in US 5,646,176), 3'-tert-butyl-3 '- N-tert-butyloxycarbonyl4-deacetyl-3'-defenyl-3'-N-debenzoyl-4-O -methoxycarbonyl-paclitaxel (described in USSN 60 / 179,965), filed February 3, 2000, incorporated herein by reference), C-4 methyl carbonate paclitaxel (described in WO 94/14787, epothilone A, epothilone B, epothilone C, epothilone D, deoxypotilone A, deoxypotilone B, [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] 7,11 -d them roxy-8,8 , 10,12,16-Pentamethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4-aza-17oxabicyclo [14.1.0] heptadecane-5,9-dione (described in WO 99/02514), F1SF1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] - 3- [2- [2- (aminomethyl) -4-thiazolyl] -1 -methylethenyl] 7,11-dihydroxy-8,8,10,12,16-pentamethyl-4,17-dioxabicyclo [14.1.0] heptadecane-5,9-dione (described in USSN 09 / 506,481, filed Feb. 17 2000, which is incorporated herein by reference), doxorubicin, carminomycin, daunorubicin, aminopterin, methotrexate, metopterin, dichloro-methotrexate, mitomics n C, porphyromycin, 5-fluorouracil, 6mercaptopurine, gemcitabine, cytosine arabinoside, podophyllotoxin or podophyllotoxin derivatives such as etoposide, etoposide phosphate or teniposide, melphalan, vinblastine, vincristine, levrosidine, vindezine, sub-inesine, vinesin Other useful anti-cancer agents that can be used in combination with the compounds of the invention include, but are not limited to, estramustine, cisplatin, carboplatin, cyclophosphamide, bleomycin, tamoxifen, ifosamide,

-2525 melphalan, hexamethyl melamine, thiotepa, cytarabine, idatrexate, trimethrexate, dacarbazine,

L-asparaginase, camptothecin, CPT-11, topotecan, ara-C, bicalutamide, flutamide, leuprolide, pyridobenzoindole derivatives, interferons, interleukins and the like. In addition, the compounds of this invention may be used in combination with farnesyl / protein / transferase inhibitors such as those described in U.S. Pat. 6,011,029; anti-angiogenic agents such as angiostatin and endostatin; kinase inhibitors such as her2-specific antibodies; and p53 transactivation modulators.

When formulated as a fixed dose, such combination products comprise the compounds of the invention within the range of doses described below and another pharmaceutically active agent to the extent of the approved dose. The compounds of formula I may be used sequentially, in any order, with known anticancer and cytotoxic agents when the combined formulation is inadequate.

The present invention also describes pharmaceutical compositions comprising a compound of the present invention and a pharmaceutically acceptable carrier. It should be emphasized that in the context of the pharmaceutical compositions of the present invention, the compounds of the invention or the compounds of formula I refer to the free base, enantiomers, diastereomers, solvates and pharmaceutically acceptable salts. Examples of such pharmaceutically acceptable salts include, but are not limited to, hydrochloride, hydrobromide, dihydrochloride, sulfate, trifluoroacetate, tartrate, fumarate, succinate, maleate, citrate, methanesulfonate, bromate and iodate salts or mixtures thereof. Also included are salts formed with organic and inorganic acids such as hydroxymethane sulfonic acid, acetic acid, benzenesulfonic acid, toluenesulfonic acid and various others, e.g. nitrates, phosphates, borates, benzoates, ascorbates, salicylates and the like. These salts include racemic forms and enantiomers and diastereomers (such as, for example, D-tartrate and L-tartrate salts). In addition, the pharmaceutically acceptable salts of the compounds of formula I may be formed with alkali metals such as sodium, potassium and lithium; alkaline earth metals such as calcium and magnesium; organic bases such as dicyclohexylamine, tributylamine and pyridines and the like; and amino acids such as arginine, lysine and the like.

The pharmaceutical compositions of the present invention may further comprise one or more pharmaceutically acceptable additional carriers, excipients or diluents, including

-2626 ingredients such as galun, stabilizers, antimicrobials, buffers, colorants, flavoring agents and the like, but not limited to. The compounds and compositions of the invention may be administered orally or parenterally, including intravenously, intramuscularly, intraperitoneally, subcutaneously, rectally or topically.

For oral administration, the compounds and compositions of the present invention may be administered, e.g. in the form of tablets or capsules or as a solution or suspension. In the case of tablets for oral use, lactose carriers and corn starch with lubricants such as magnesium stearate are commonly used. For oral administration, capsules are used as useful lactose carriers and corn starch. When aqueous suspensions are used for oral administration, emulsifying and / or suspending agents are often added. In addition, sweeteners and / or flavoring agents may be added to the oral compositions. For intramuscular, intraperitoneal, subcutaneous and intravenous use, sterile solutions of the active ingredients are usually used, the pH of the solutions should be adjusted appropriately and buffered. For intravenous administration, the total solute concentration must be controlled to make the preparation isotonic.

The daily doses for human administration of the compounds of the invention will typically be determined by the treating physician, with dosages generally varying according to the age, weight, route of administration and response of the individual patient, and the severity of the patient's symptoms. The compound of formula I of the present invention is preferably administered to humans in an amount of from about 0.001 mg / kg body weight to about 100 mg / kg body weight per day, more preferably from about 0.01 mg / kg body weight to about 50 mg / kg body weight per day and more preferably from about 0.1 mg / kg body weight to about 20 mg / kg body weight per day.

Cdc2 / cyclin B1 kinase assay

Cdc2 / cyclin B1 kinase activity was determined by monitoring the attachment of ³² P into histone HI. The reaction consisted of 50 ng of baculovirus-expressed GST-cdc2, 75 ng of baculovirus-expressed GST-cyclin B1, 1 pg of histone HI (Boehringer Mannheim), 0.2 μθΐ ³² P γ-ΑΤΡ, and 25 μΜ ATP in kinase buffer (50 mM Tris, pH 8.0, 10 mM MgCb, 1 mM EGTA, 0.5 mM DTT). The reaction was incubated at 30 ° C for 30 minutes and then quenched with the addition of cold trichloroacetic acid (TCA) to a final concentration of 15% and incubated on ice for 20 minutes. The reaction was picked up on GF / C unifilter plates (Packard)

-2727 using a Packard Filtermate Universal picker and count the filters on a 96-well Packard TopCount liquid scintillation counter (Marshak, DR, Vanderberg, MT, Bae, YS, Yu, JJ :, J. of Cellular Biochemistry, 45, 391-400 ( 1991) which is incorporated by reference).

Cdk2 / cyclin E kinase assay

Cdk2 / cyclin E kinase activity was determined by monitoring the incorporation of ³² P into the retinoblastoma protein. The reaction consisted of 2.5 ng of baculovirus-expressed GSTcdk2 / cyclin E, 500 ng of bacterially produced GST-retinoblastoma protein (aa 776-928), 0.2 μθϊ ³² P γ-ΑΤΡ and 25 μΜ ATP in kinase buffer (50 mM Hepes , pH 8.0, 10 mM MgCb, 5 mM EGTA, 2 mM DTT). The reaction was incubated at 30 ° C for 30 minutes and then quenched with the addition of cold trichloroacetic acid (TCA) to a final concentration of 15% and incubated on ice for 20 minutes. The reaction was collected on GF / C unifilter plates (Packard) using a Packard Filtermate Universal picker and filters were counted on a 96 Packard TopCount liquid scintillation counter.

Cdk4 / cyclin D1 kinase activity

Cdk4 / cyclin D1 kinase activity was determined by monitoring the incorporation of ³² P into the retinoblastoma protein. The reaction consisted of 165 ng of baculovirus-expressed GST-cdk4, 282 ng bacterially expressed as S-tag cyclin D1, 500 ng of bacterially produced GST-retinoblastoma protein (aa 776-928), 0.2 gCi ³² P γ-ΑΤΡ and 25 μΜ ATP in kinase buffer (50 mM Hepes, pH 8.0, 10 mM MgCI ₂ , 5 mM EGTA, 2 mM DTT). The reaction was incubated at 30 ° C for 1 hour and then quenched by the addition of cold trichloroacetic acid (TCA) to a final concentration of 15% and incubated on ice for 20 minutes. The reaction was collected on GF / C unifilter plates (Packard) using a Packard Filtermate Universal picker and filters were counted on a 96-well Packard TopCount liquid scintillation counter (Coleman, KG, Vautaut, BS, Morissey, D, Mulheron, JG, Sedman, S ., Brinkley, P., Witnesses, S. Webster, K.R. (1997) Identification of CDK4 Sequences involved in cyclin D, and p16 binding. J. Biol. Chem. 272, 30: 18869-18874, to be incorporated by reference.

-2828

In order to facilitate further understanding of the invention, the following examples will be presented primarily to illustrate the specific compounds of the invention. The scope of the present invention is not limited by the examples, but includes the whole substance defined in the claims.

EXAMPLE 1: Preparation of 5- [5- (t-Butyl) -2-oxazolylmethylthio] -2 (azacycloalkanoyl) amino-thiazole hydrochloride

NH. HCI

A. Preparation of α-azido-pinacolone

α-Bromo-pinacolone (199.07 g, 1.1115 mol, 1 eq) was combined in 1,785 I acetone with sodium azide (93.9 g, 1.444 mol, 1.3 eq). The reaction was stirred at room temperature

27,5 hours The resulting suspension was filtered and washed with acetone (3 x 150 ml). The filtrate was concentrated in vacuo to give 154.3 g (98.4%) of the title compound. HPLC 83.85% at 2.57 min (Phenomenex Inc., Torrance, CA, 5 μιτι C18 column 4.6 x 50 mm, 10-90% aqueous methanol for 4 minutes containing 0.2% phosphoric acid , 4 ml / min, observation at 220 nm).

B. Preparation of g-hexamethylenetetramino-pinacolone bromide

Br '? -Bromo-pinacolone (179 g, 1 mol, 1 eq) was combined in 2 I acetone with hexamethylenetetramine (154.21 g, 1.1 mol, 1.1 eq) and stirred under N ₂ at room temperature.

-2929 temperature for 26 hours. The resulting suspension was filtered, the Quick Cakes washed with ether (3 x 50 ml) and dried in vacuo at 50 ° C overnight to give 330 g (100%) of the title compound containing 7% hexamethylenetetramine. HPLC ST = 0.17 min (Phenomenex Inc., 5 gm C18 column 4.6 x 50 mm, 10-90% aqueous methanol for 4 minutes containing 0.2% phosphoric acid, 4 ml / min, observation at 220 nm).

C. Preparation of α-amino-pinacolone hydrochloride

NH ₂ · HCl α-azido-pinacolone (128.5 g, 0.911 mol) was combined in 4.2 I methanol with 77.1 ml of concentrated HCI and 15.42 g of 10% Pd / C. The reaction mixture was stirred under hydrogen for 1.5 hours. The catalyst was removed by filtration. The solvent was distilled to give a wet solid. The remaining water was azeotropically removed with isopropanol (2 x 500 ml). Tert-butyl methyl ether (300 ml) was added and the resulting suspension was stirred, filtered, washed with t-butyl methyl ether (3 x 100 ml) and dried to give 131.0 g (95.5) %) of the title compound.

D. Preparation of α-amino-pinacolone hydrochloride

O \ A / NH ₂ · HCl α-hexamethylenetetramino-pinacolone bromide (400 g, 1.254 mol, 1 eq) was combined in 2 I of ethanol with 12 N aqueous HCl (439 ml, 5.26 mol, 4.2 eq). The reaction was stirred at 75 ° C for 1 hour and then cooled to room temperature, the resulting suspension was filtered, the filtrate was concentrated in vacuo and isopropyl alcohol was added. The solution was filtered again. The addition of 1.2 I ether caused the desired material to precipitate from the solution. The material was filtered, washed with ether (2 x 300 ml) and dried in vacuo at 50 ° C overnight to give 184.1 g (97%) of the title compound.

-3030

E. Preparation of g-N- (2-chloroacetylamino) -pinacolone o

The title compound of part D (130.96 g, 0.8637 mol, 1 eq) was dissolved in 3.025 I CH2Cl2 under N2 at -5 ° C. Triethylamine (301 ml, 2.16 mol, 2.5 eq) was then added, then chloroacetyl chloride (15.7 ml, 0.450 mol, 1.1 eq) in 175 ml of CH 2 Cl 2 · The resulting suspension was stirred at -5 to -10 ° C for 2 hours. Water (1.575 I) and 175 ml of concentrated HCl were added. The organic phase was washed a second time with 1.75 I of 10% aqueous HCl and then with 500 ml of water. The organic phase was dried over Na ₂ SO ₄ and concentrated in vacuo to give

155.26 g (93.8%) of the title compound. HPLC ST = 2.27 min (Phenomenex Inc., 5 pm C18 column 4.6 x 50 mm, 10-90% aqueous methanol for 4 minutes containing 0.2% phosphoric acid, 4 ml / min, observation at 220 nm).

F. Preparation of 5- (t-butyl) -2-oxazolylmethyl chloride

The title compound of part E (180.13 g, 0.9398 mol, 1 eq) was combined with phosphorus oxychloride (262 ml, 2.8109 mol, 3 eq) under N ₂ . The reaction was heated at 105 ° C for 1 hour, cooled to room temperature and quenched with 1.3 kg of ice. The aqueous phase was extracted with ethyl acetate (1 L, then 2 x 500 ml). The organic extracts were washed with saturated aqueous NaHCO ₃ (4 X 1 I), which was back extracted several times with ethyl acetate. The organic phases were combined, washed with saturated aqueous NaHCO ₃ (500 ml) and saturated aqueous NaCl (300 ml), dried over MgSO ₄ and concentrated in vacuo to give a brown oil. The crude material was distilled under high vacuum at 100 ° C to give 155.92 g (96%) of the title compound. HPLC ST = 3.62 min (Phenomenex Inc., 5 μηι C18 column 4.6 x 50 mm, 10-90% aqueous methanol for 4 minutes containing 0.2% phosphoric acid, 4 ml / min, observation at 220 nm).

-3131

Alternatively, the title compound of part E (10.0 g, 52.17 mmol, 1 eq) in 50 ml of tetrahydrofuran (THF) was combined with (methoxycarbonylsulfamyl) -triethylammonium hydroxide (Burgess reagent, 105.70 mmol, 2.03 eq. generated in situ from 9.2 ml of chlorosulfonyl isocyanate, 4.4 ml of methanol and 14.8 ml of triethylamine in 100 ml of THF). The reaction was heated to 45 ° C for 1.5 hours. After cooling to room temperature, the reaction was quenched with water (50 ml). The organic layer was separated and washed with saturated NaHCO3 (2 x 50 ml) and water (50 ml), dried over MgSO ₄ and passed through a small silica gel stopper. The solvent was removed to give an oil which was introduced into a mixture of 15 ml of heptane and 90 ml of tbutyl methyl ether and then washed with 0.2 N HCl (2 x 25 ml), saturated brine (25 ml) and dried. (MgSO ₄ ). Filtration and solvent removal gave 10.9 g of the title compound.

G. Preparation of 5- (t-butyl) -2-oxazolylmethyl thiouronium hydrochloride

The title compound of part F (1.77 g, 10.2 mmol, 1.02 eq) was combined with a thiourea (0.76 g, 9.98 mmol, 1 eq) under N ₂ in 10 ml absolute ethanol. The reaction was heated at reflux for 1.5 hours. The mixture was cooled to room temperature and concentrated in vacuo. Trituration of the crude material obtained with t-butyl methyl ether gave 2.32 g (93%) of the title compound. HPLC ST = 2.05 min (Phenomenex Inc., 5 pm C18 column 4.6 x 50 mm, 10-90% aqueous methanol for 4 minutes containing 0.2% phosphoric acid, 4 ml / min, observation at 220 nm); ¹ H NMR (d ₆ -DMSO); δ 9.48 (s, 3H), 6.85 (s, 1H), 4.73 (s, 2H), 1.24 (s, 9H).

H. Preparation of 5- | ~ 5- (t-Butyl) -2-oxazolylmethylthio1-2-aminothiazole

-3232

The title compound of part G (1.25 g, 5 mmol, 1 eq) was added to a mixture of NaOH (3.0 g, mmol, 15 eq), water (10 ml), toluene (10 ml) and tetrabutylammonium sulfate (50 mg , 0.086 mmol, 0.017 eq). 5-Bromo-2-aminothiazole hydrobromide (1.70 g, 5 mmol, 1 eq) was added and the reaction was stirred at room temperature for 14.5 hours. The mixture was diluted with water and extracted twice with ethyl acetate, the organic extracts were washed with water (4x10 ml), dried over MgSO ₄ and concentrated in vacuo to give 1.1 g (82%) of the title compound. HPLC 86.3% 2.75 min (Phenomenex Inc., 5 μίτι C18 column 4.6 x 50 mm, 1090% aqueous methanol for 4 minutes containing 0.2% phosphoric acid, 4 ml / min, observation at 220 nm); ¹ H NMR (CDCl 3): δ 6.97 (s, 1H), 6.59 (s, 1H), 5.40 (br s, 2H), 3.89 (s, 2H), 1.27 (s , 9H).

I. Preparation of 5- [5- (t-Butyl) -2-oxazolylmethylthio1-2 - [(N-t-butoxycarbonyl) azacycloalkanoylaminothiazole

N S

o

The title compound of part H (9.6 g, 35.6 mmol) was dissolved in N, N-dimethylformamide (36 ml) and CH2Cl2 (100 ml) to which 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride was added. (13.8 g, 72 mmol, 2 eq), Nt-butoxycarbonyl-azacycloalkanoic acid (12.6 g, 55 mmol, 1.5 eq) and 4- (dimethylamino) pyridine (2 g, 16 mmol, 0.45 ek). The clear reaction mixture became cloudy when stirred at room temperature for 3.5 hours. Water (300 ml) and ethyl acetate (200 ml) were added and the resulting precipitate was removed by filtration. The filtrate was extracted with ethyl acetate, the organic extracts were dried over MgSO ₄ and concentrated in vacuo to give a yellow solid, which was combined with a precipitate obtained by filtration. The solid was boiled in a mixture of ethanol, acetone and water for 20 minutes, filtered, washed with a mixture of ethanol and water and dried to give 16.6 g (97%) of the title compound.

J. Preparation of 5- [5- (t-Butyl) -2-oxazolylmethylthioyl-2- (azacycloalkanoyl) aminothiazole hydrochloride

-3333 / PU n

The title compound of part I (16.6 g) was dissolved in 150 ml of CH2Cl2, trifluoroacetic acid (30 ml) was added dropwise and the mixture was stirred at room temperature for 2 hours. The reaction was concentrated in vacuo, diluted with water (300 ml), cooled on ice, made basic with sodium hydroxide and the resulting solid filtered and recrystallized from ethanol, water and methanol to give 11.2 g (83%) of the title compounds as a yellow solid. White solid hydrochloride could be obtained by adding 18 ml of 1N aqueous HCl in 7 g of this material in methanol. MS: 381 [M + H] &lt; + &^gt;; HPLC: 100% at 3.12 min (YMC S5 ODS column 4.6 x 50 mm, 10-90% aqueous methanol for 4 minutes containing 0.2% phosphoric acid, 4 ml / min, observation at 220 nm).

Example 2: Preparation of (±) -N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methylthiol-2-thiazolyl] -3-piperidinecarboxamide

A. (±) -N-t-Butoxycarbonyl-nipecinic acid

N

BOC

Ni-nicotinic acid (1.3 g, 10 mmol, 1 eq) was combined with 10 ml of dioxane, 2 ml of acetonitrile, 10 ml of water and 10 ml of 1N aqueous NaOH (1 eq). Di-p-butyldicarbonate (3.3 g, 15 mmol, 1.5 eq) was added and the reaction mixture was stirred at rt overnight. The reaction mixture was concentrated in vacuo to remove the organic solvent and

-3434 added 10% aqueous citric acid. The mixture was extracted with ethyl acetate (3 x 100 ml). The organic extracts were dried over Na ₂ SO ₄ , filtered through silica gel and concentrated in vacuo. The crude material was recrystallized from ethyl acetate and hexanes to give 2.2 g (96%) of (±) -Nf-butoxycarbonyl-nicotinic acid as a white solid.

B. (±) -N-f5-ii [5- (1,1-dimethylethyl) -2-oxazolylmethylthio] -2-thiazolyl 1- (N-butoxycarbonyl) -3-piperidinecarboxamide

1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (383 mg, 2 mmol, 2 eq) was added to a mixture of 2-amino-5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] thiazole (270 mg, 1 mmol, 1 eq), Nf-butoxycarbonyl-nipecic acid (344 mg, 1.5 mmol, 1.5 eq), 4 (dimethylamino) pyridine (61 mg, 0.5 mmol, 0 , 5 eq), N, N-dimethylformamide (1 ml) and CH ₂ Cl ₂ (6 ml). The reaction mixture was stirred at st 1.3 hours. Triethylamine (0.28 mL, 2 mmol, 2 eq) was added and the reaction mixture was stirred for 1 h. Additional Nf-butoxycarbonylpecotinic acid (340 mg), triethylamine (0.28 ml) and 1- (3-dimethylaminopropyl) -3ethylcarbodiimide hydrochloride (380 mg) were added. No further changes were observed after 1 h. 4- (dimethylamino) pyridine, N, N-dimethylformamide, triethylamine and starting acids were added and the reaction stirred overnight at rt. The resulting black solution was diluted with saturated aqueous NaHCCL and extracted with CH ₂ Cl ₂ . The organic extracts were dried, concentrated in vacuo and purified by flash chromatography on silica gel eluting with a gradient of 50-100% ethyl acetate in hexanes to give 397 mg (83%) (±) -N- [5- [ [[5 (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] - (Nf-butoxycarbonyl) -3piperidinecarboxamide as a yellow glassy solid.

C. Preparation of (±) -N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolylmethyl] thio] -2-thiazolyl-3-piperidinecarboxamide

-3535

(±) -N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] - (Nt-butoxycarbonyl) -3piperidinecarboxamide (355 mg, 0.74 mmol, 1 eq) was dissolved in 3 ml of CH ₂ Cl ₂ . Trifluoroacetic acid (3 ml) was added and the mixture was stirred at 20 min. The reaction mixture was concentrated in vacuo and neutralized with saturated aqueous NaHCO ₃ . The resulting mixture was extracted with ethyl acetate. The organic extracts were dried over Na ₂ SO ₄ , concentrated in vacuo and recrystallized from ethyl acetate to give 142 mg (50%) (±) -N- [5 - [[[5- (1,1-dimethylethyl) ) -2-oxazolyl] methyl] thio] -2-thiazolyl] -3-piperidinecarboxamide as a white solid. MS: 381 [M + H] &lt; + &^gt;; HPLC: 100% at 3.15 min (YMC S5 ODS column 4.6 x 50 mm, 10-90% aqueous methanol for 4 minutes containing 0.2% phosphoric acid, 4 ml / min, observation at 220 nm).

Example 3: Preparation of (±) -1- (2,3-dihydroxypropyl) -N- [5 [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] 2-thiazolyl 1-4-piperidinecarboxamide

N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide (66 mg, 0.17 mmol, 1 equiv) were combined with glyceraldehyde (69 mg, 0.77 mmol, 4.5 eq), sodium triacetoxyborohydride (163 mg, 0.77 mmol, 4.5 eq) and 1,2-dichloroethane (4 ml). The resulting suspension was stirred at 4 h. Methanol (1 ml) was added and the reaction mixture was stirred at rt overnight, concentrated in vacuo and purified by preparative HPLC to afford 69 mg (59%) (±) -1- (2,3-dihydroxypropyl) -N - [5 - [[[5- (1,1-dimethylethyl) -2oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide as a white solid. MS: 455 [M + H] &lt; + &^gt;; HPLC: 100% at 3.06 min (YMC S5 ODS column 4.6 x 50 mm, 10-90%

-3636 aqueous methanol for 4 minutes containing 0.2% phosphoric acid, 4 ml / min, observation at 220 nm).

Example 4: Preparation of N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl-1- (1-methylethyl) -4-piperidinecarboxamide

A. Ethyl-1- (1-methylethyl) -4-piperidine carboxylate

COaEt

Ethyl isonipecotate (3.2 g, 20 mmol, 1 eq) was combined with acetone (5.8 g, 100 mmol, 5 eq), sodium triacetoxyborohydride (10.5 g, 50 mmol, 2.5 eq) and 1. 2-dichloroethane (200 ml). The reaction mixture was stirred at 72 h. Saturated aqueous NaHCO ₃ was added and the mixture was extracted with CH ₂ Cl ₂ . The organic extracts were dried, filtered through a silica gel sponge, and concentrated in vacuo to give 3.72 g (93%) of ethyl 1- (1methylethyl) -4-piperidine carboxylate as a colorless liquid.

B. 1- (1-Methylethyl) -4-piperidine carboxylic acid

-3737 co ₂ hrs

Ethyl 1- (1-methylethyl) -4-piperidine carboxylate (3.6 g, 18 mmol, 1 eq) was combined with barium hydroxide octahydrate (10.4 g, 33 mmol, 1.8 eq) in a mixture of 70 ml of water with 44 ml of ethanol. The mixture was heated at 60 ° C for 1.3 h. The reaction mixture was concentrated in vacuo and diluted with 70 ml of water. Ammonium carbonate (6.9 g, 87 mmol, 4.8 eq) was added portionwise and the reaction mixture was stirred at rt overnight. The mixture was filtered through diatomaceous earth, concentrated and lyophilized to give 3.1 g (100%) of 1- (1methylethyl) -4-piperidine carboxylic acid as a white solid.

C. N- [5- [| T5- (1,1-dimethylethyl) -2-oxazolylmethylthio1-2-thiazolyl] -1- (1 methylethyl) -4-piperidinecarboxamide

1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (1.0 g, 5.2 mmol, 2 eq) was added to a mixture of 2-amino-5 - [[[5- (1,1-dimethylethyl) -2- oxazolyl] methyl] thio] thiazole (0.7 g, 2.6 mmol, 1 eq), 1- (1-methylethyl) -4-piperidine carboxylic acid (0.78 g, 3.9 mmol, 1.5 eq) ), 4 (dimethylamino) pyridine (0.16 g, 1.3 mmol, 0.5 eq), N, N-dimethylformamide (2.6 ml) and CH2Cl2 (7.8 ml). The reaction mixture was stirred at rt for 1 h, diluted with 30 ml water and extracted with ethyl acetate (2 x 70 ml). The organic extracts were dried over Na ₂ SO ₄ , concentrated in vacuo and purified by flash chromatography on silica gel eluting with a gradient of 5-10% triethylamine in ethyl acetate. The material was recrystallized from ethanol and

-3838 water to give 0.93 g (85%) of N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -1- (1- methylethyl) -4-piperidinecarboxamide as a yellowish solid. MS: 423 [M + H] &lt; + &^gt;; HPLC: 100% at 3.15 min (YMC S5 ODS column 4.6 x 50 mm, 10-90% aqueous methanol for 4 minutes containing 0.2% phosphoric acid, 4 ml / min, observation at 220 nm).

Example 5: Preparation of 1-cyclopropyl-N- [5 - [[[5- (1,1-dimethylethyl) -2oxazolyl] methylthiol-2-thiazoline-4-piperidinecarboxamide

A. 1-Cyclopropyl-4-piperidine carboxylic acid co ₂ h

N '

Ethyl isonipecotate (1.57 g, 10 mmol, 1 eq) was combined with ((1ethoxycyclopropyl) oxy) trimethyl Silane (8.7 g, 50 mmol, 5 eq) in 100 ml of methanol. Acetic acid (5.7 ml, 100 mmol, 10 eq) and molecular sieves were added. After 30 min at rt, sodium triacetoxyborohydride (2.5 g, 40 mmol, 4 eq) was added and the reaction mixture was heated at 65 ° C overnight. The reaction mixture was cooled and Na2CO ₃ (20 g). The mixture was stirred at st 2 h and filtered through diatomaceous earth. The diatomaceous earth was washed with methanol. The filtrates were combined, concentrated in vacuo, diluted with water and extracted with ethyl acetate. The organic extracts were dried, filtered through a silica gel sponge and concentrated in vacuo to give 2.4 g of a colorless liquid. This material was combined with barium hydroxide octahydrate (5.7 g,

-3939 mmol, 1.8 eq) in a mixture of 38 ml of water with 24 ml of ethanol. The mixture was heated at

60 ° C 1 h. The reaction mixture was concentrated in vacuo and diluted with 38 ml of water. Ammonium carbonate (3.8 g) was added portionwise and the reaction stirred at rt for 2 h. The mixture was filtered through diatomaceous earth, washed with water. The filtrate was washed with ethyl acetate. The aqueous phase concentration gave 1.56 g (92%) of 1-cyclopropyl-4-piperidine carboxylic acid as a hygroscopic white solid.

B. 1-cyclopropyl-N- [5- [f [- (1,1-dimethylethyl) -2-oxazolyl-1-methythioi-2-thiazolyl] 4-piperidinecarboxamide

1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (1.0 g, 5.2 mmol, 2 eq) was added to a mixture of 2-amino-5 - [[[5- (1,1-dimethylethyl) -2- oxazolylmethyl] thiothiazole (0.7 g, 2.6 mmol, 1 eq), 1-cyclopropyl-4-piperidine carboxylic acid (0.77 g, 3.9 mmol, 1.5 eq), 4 (dimethylamino) pyridine ( 0.16 g, 1.3 mmol, 0.5 eq), N, N-dimethylformamide (2.6 ml) and CH ₂ Cl ₂ (7.8 ml). The reaction mixture was stirred at rt for 1 h, diluted with water (30 ml) and extracted with ethyl acetate (2 x 70 ml). The combined organic extracts were dried over anhydrous sodium sulfate, concentrated in vacuo and purified by flash chromatography on silica gel eluting with a 0-10% triethylamine gradient in ethyl acetate. The material was crystallized from ethyl acetate and hexanes to give 0.7 g (65%) of 1-cyclopropyl-N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] - 2-Thiazolyl] -4-piperidinecarboxamide as white crystals. MS: 421 [M + H] &lt; + &^gt;; HPLC: 100% at 3.13 min (YMC S5 ODS column 4.6 x 50 mm, 10-90% aqueous methanol for 4 minutes containing 0.2% phosphoric acid, 4 ml / min, observation at 220 nm).

Example 6: Preparation of N- [5 - [[[5- (1,1-dimethylethyl) -oxazolylmethylthio] -thiazolyl-1- (4-hydroxyethyl) -piperidinecarboxamide

-4040

A. N-f5- [ff5- (1,1-dimethylethyl) -2-oxazolinmethyl] thio1-2-thiazoline-1- (2-dimethyl-butylsilyloxyethyl) -4-piperidinecarboxamide

N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide (1.4 g,

3.68 mmol, 1 equiv) was dissolved in 30 ml of Ν, Ν-dimethylformamide and 100 ml of tetrahydrofuran. 2- (Bromoethoxy) -f-butyldimethylsilane (0.79 ml, 3.68 mmol, 1 eq) and NaHCO3 were added and the reaction mixture was stirred at 50 ° C for 23 h. Additional 2- (bromoethoxy) -butyldimethylsilane (0.9 ml) was added and the reaction mixture was stirred at 50 ° C for 22 h, cooled, concentrated in vacuo and diluted with water (25 ml). The resulting aqueous mixture was extracted with ethyl acetate (50 ml). The organic extract was dried over Na ₂ SO ₄ , concentrated in vacuo and purified by flash chromatography on silica gel eluting with a gradient of 0-5% triethylamine in ethyl acetate to give 1.7 g (84%) - [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -1- (2-dimethyl-1-butylsilyloxyethyl) -4-piperidinecarboxamide as a yellow solid. MS: 539 [M + H] +; HPLC: 98% at 4.01 min (YMC S5 ODS column 4.6 x 50 mm, 10-90% aqueous methanol for 4 minutes containing 0.2% phosphoric acid, 4 ml / min, observation at 220 nm).

B. N-f5- [ff5- (1,1-dimethylethyl) -2-oxazolylmethylthioyl-2-thiazolin-1- (2hydroxyethyl) -4-piperidinecarboxamide

-4141 η

(CH ₃ ) ₃ cr

N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -1- (2-dimethyl-t-butylsilyloxyethyl) -4-piperidinecarboxamide (1.45 g , 2.7 mmol, 1 eq) was dissolved in 100 ml of acetonitrile and combined with aqueous HF (48% aqueous, 2.5 ml). The reaction mixture was stirred for 4 h at st. An additional 2.5 ml of aqueous HF was added and the reaction mixture was stirred overnight. Ethyl acetate (100 ml) and saturated aqueous NaHCO3 (50 ml) were added. Additional solid NaHCO ₃ was added to make the mixture basic. The mixture was extracted with ethyl acetate (2 x 50 ml). The organic extracts were dried over Na ₂ SO ₄ , filtered through a silica gel sponge and concentrated in vacuo. The resulting white solid was crystallized from ethanol and water to give 1.6 g (59%) of N- [5 - [[[5- (1,1-dimethylethyl) -2oxazolyl] methyl] thio] -2-thiazolyl ] -1- (2-hydroxyethyl) -4-piperidinecarboxamide as a white solid. MS: 425 [M + H] &lt; + &^gt;; HPLC: 100% at 3.05 min (YMC S5 ODS column 4.6 x 50 mm, 10-90% aqueous methanol for 4 minutes containing 0.2% phosphoric acid, 4 mi / min, observation at 220 nm).

Example 7: Preparation of (R) - [5 - [[[5- (1,1-dimethylethyl) -2oxazolyl] methyl] thio] -2-thiazolyl] -3-piperidinecarboxamide hydrochloride

A. (R) - and (S) -N- [5-r [[5- (1,1-dimethylethyl) -2-oxazoylmethyl] thio] -2-thiazolyl- (Nf-butoxycarbonyl) -3-piperidinecarboxamide

-4242

(R)

(S)

1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (3.8 g, 20 mmol, 2 eq) was added to a mixture of 2-amino-5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] thiazole (2.7 g, 10 mmol, 1 eq), Ν-ί-butoxycarbonyl-nicotinic acid (3.4 g, 1.5 mmol, 1.5 eq), N, N-dimethylformamide (10 ml) and CH2Cl2 (30 ml). The reaction mixture was stirred at 4 h. The resulting black solution was concentrated in vacuo, diluted with water (90 ml) and extracted with ethyl acetate (100 ml then 2 x 75 ml). The organic extracts were dried over Na2CO3, concentrated in vacuo and purified by flash chromatography on silica gel eluting with a gradient of 50-100% ethyl acetate in hexanes to give 3.8 g (79%) as a yellow solid. The enantiomers were separated by chiral HPLC (Chiral Pak AD 5 X 50 cm 20 μ: eluent 10% (0.1% triethylamine in isopropanol) in hexanes; 45 ml / min, detection at 254 nm, load 300 mg in 5 ml isopropanol ) to give each of two optically pure isomers: 1.65 g of the R isomer and 1.65 g of the S isomer.

B. (R) -N- [5-i [[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thioj-2-thiazolyl] -3-piperidinecarboxamide hydrochloride

The -4343 (R) isomer of part A (1.65 g, 3.43 mmol, 1 eq) was dissolved in 10 ml of CH 2 Cl 2. Trifluoroacetic acid (6 ml) was added and the mixture was stirred for several hours. The reaction mixture was concentrated in vacuo and neutralized with saturated aqueous NaHCO ₃ . The resulting mixture was stirred with ethyl acetate for 1 h. The organic extracts were dried over Na2SO ₄ and concentrated in vacuo to provide a yellowish solid. The solid was dissolved in methanol and 1 eq of 1 N aqueous HCl was added. The resulting solution was lyophilized to give 1 g (77%) of (R) -N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] - 3piperidinecarboxamide hydrochloride as a yellow solid. MS: 381 [M + H] &lt; + &^gt;; HPLC: 100% at 3.14 min (YMC S5 ODS column 4.6 x 50 mm, 10-90% aqueous methanol for 4 minutes containing 0.2% phosphoric acid, 4 ml / min, observation at 220 nm).

Example 8: Preparation of (S) -N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl-3-piperidine carboxamide hydrochloride

(S) The isomer of Example 7, Part A (1.65 g, 3.43 mmol, 1 eq) was dissolved in 10 ml of CH 2 Cl 2. Trifluoroacetic acid (6 ml) was added and the mixture was stirred for several hours. The reaction mixture was concentrated in vacuo and neutralized with saturated aqueous NaHCO ₃ . The resulting mixture was stirred with ethyl acetate for 1 h. The organic extracts were dried over Na2SO ₄ and concentrated in vacuo to provide a yellowish solid. The solid was dissolved in methanol and 1 eq of 1 N aqueous HCl was added. The resulting solution was lyophilized to give 0.918 g (70%) of (S) -N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] 2-thiazolyl] -3 -piperidinecarboxamide hydrochloride as a yellow solid. MS: 381 [M + H] &lt; + &^gt;; HPLC: 100% at 3.15 min (YMC S5 ODS column 4.6 x 50 mm, 10-90% aqueous methanol for 4 minutes containing 0.2% phosphoric acid, 4 ml / min, observation at 220 nm).

-4444

Example 9: Preparation of cis-4-amino-N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] cyclohexylcarboxamide hydrochloride and trans-4 -amino- N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] -methyl] thio] -2-thiazolylcyclohexylcarboxamide hydrochloride

A. 4- (f-Butoxycarbonylamino) cyclohexane carboxylic acid ^ NHfBOC

KO ₂ C

A total of 6.5 g (30 mmol) (Boc of anhydride at room temperature) was added to a solution of 2.86 g (20 mmol) of 4-aminocyclohexane carboxylic acid in 40 ml of 0.5 M aqueous NaOH, 20 ml of dioxane and 4 ml of acetonitrile. After 20 h, 100 ml of ethyl acetate and 100 ml of a solution of 10% aqueous citric acid were added and the resulting aqueous layer was separated and extracted with three 50 ml portions of ethyl acetate. The organic phases were combined, dried (sodium sulfate) and concentrated in vacuo to give 6.0 g (125%) of crude 4- (fbutoxycarbonylamino) cyclohexane carboxylic acid as a colorless oil which solidified upon rest.

B.

4- (t-Butoxycarbonylamino) -N-f5-r [[- (1,1-dimethylethyl) -2-oxazolyl] -methyl] thio-

NH

NHffiOC

-4545

A solution of 5 g of crude 4- (t-butoxycarbonylamino) cyclohexane carboxylic acid and 3.50 g (13 mmol) of 2-amino-5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] of thiazole in 13 ml of N, N-dimethylformamide and 36 ml of methylene chloride were added 5.0 g (26 mmol) of 1- (3dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride at room temperature. The reaction mixture was stirred overnight and diluted with 100 ml of water. The aqueous layer was separated and extracted with two 150 mL portions of ethyl acetate. The combined organic phases were dried (sodium sulfate) and then filtered through a silica gel sponge. The filtrate was concentrated in vacuo to give an orange solid. The crude material was recrystallized (95% ethanol) to give 4- (f-butoxycarbonylamino) -N- [5 - [[[- (1,1-dimethylethyl) -2-oxazolyl] methyl] thioj-2-thiazolyl] cyclohexylcarboxamide as a yellow solid. The mother liquors were also concentrated in vacuo to give additional 4- (butoxycarbonylamino) -N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolylcyclohexylcarboxamide as a brown solid .

C. cis-4-Amino-N-pyrrolidine- (1,1-dimethylethyl) -2-oxazolyl-1-methylthio12-thiazolylcyclohexylcarboxamide hydrochloride and trans ~ 4 amino- N- [5-ff [5- (1,1 -dimethylethyl) -2-oxazolyl1-methynthio-2-

Suspensions of 4- (t-butoxycarbonylamino) -N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolylcyclohexylcarboxamide (from Part B of mother liquors) suspended in 15 ml of methylene -chloride, 5 ml of trifluoroacetic acid were added at room temperature. The reaction mixture was stirred for 2 h, then concentrated in vacuo to remove volatiles. The residue was diluted with water, made basic with aqueous NaOH solution and

-4646 was then extracted with the aqueous solution with ethyl acetate. The combined organic extracts were dried (sodium sulfate) to give the crude cis / trans product. The crude material was purified by flash chromatography (Merck silica, 25x3 cm, 1: 9 isopropylamine / ethyl acetate, then 1: 2: 7 methanol / isopropylamine / ethyl acetate to give 0.74 g of the cis isomer as a yellow solid and 0 , 50 g of the trans isomer as a brown solid The cis isomer was dissolved in methanol and then 0.34 ml of 5 N aqueous HCl was added, the solution was concentrated in vacuo, washed with ether, diluted with water and lyophilized to give 0. 80 g of cis-4-amino-N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolylcyclohexylcarboxamide hydrochloride as a yellow solid MS: 395 [M + H] ⁺ ; HPLC: 98% at 3.17 min (YMC S5 ODS column 4.6 x 50 mm, 10-90% aqueous methanol for 4 minutes containing 0.2% phosphoric acid, 4 ml / min, observation The trans isomer was dissolved in methanol and then 0.24 ml of 5N aqueous HCl was added, the solution was concentrated in vacuo, washed with ether, diluted with water and lyophilized to give 0.54 g of trans-4- amino-N- [5 - [[[5- (1,1-dim thylethyl) -2oxazolyl] methyl] thio] -2-thiazolyl] cyclohexylcarboxamide hydrochloride as an orange solid. MS: 395 [M + H] &lt; + &^gt;; HPLC: 96% at 3.22 min (YMC S5 ODS column 4.6 x 50 mm, 10-90% aqueous methanol for 4 minutes containing 0.2% phosphoric acid, 4 ml / min, observation at 220 nm) .

Example 10: N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide, monohydrochloride

Acetyl chloride (0.28 ml, 3.9 mmol) was added dropwise to a solution of 40 ml of absolute EtOH cooled in an ice bath. The reaction mixture was warmed to room temperature for 30 min and then N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide was added in one portion. (1.50 g, 3.94 mmol, 1 eq) while stirring to obtain a thick suspension. Water (~ 4 ml) was added to give a homogeneous mixture and then concentrated in vacuo to give a crude pale yellow solid. The crude material was recrystallized (aq. EtOH) to give the title compound (70%) as white

-4747 solid, m.p. 256-258 ° C. Analysis calculated for C17H24N4O2S2HCI: C, 48.96; H,

6.04; N, 13.43; S, 15.38; Cl, 8.50. Found: C, 48.69; H, 5.99; N, 13.24; S, 15.27; Cl,

8.31.

Example 11: N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide, monohydrobromide

To a solution of 1M HBr in EtOH (0.5 ml) may be added N- [5 - [[[5- (1,1-dimethylethyl) -2oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide (190 mg, 0.5 mmol, 1 equiv) was then cooled to -40 ° C overnight. The resulting solid precipitate was collected on a Buchner funnel, washed with absolute EtOH, then dried under vacuum at 100 ° C to give the title compound (72%) as a fine white powder, m.p. Mp 235-237 ° C. Analysis calculated for C17H24N4O2S2HBr: C, 44.24; H, 5.46; N, 12.14; S, 13.89; Br, 17.31. Found: C, 44.16; H, 5.40; N, 12.12; S, 13.91; Br, 17.70.

Example 12: N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide, 0,5-L-tartrate

Warm solutions of N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide (1.75 g, 4.6 mmol) in absolute EtOH ( 70 ml) was added a solution of L-tartaric acid (345 mg, 2.3 mmol, 0.5 eq) in absolute EtOH (5 ml). After a few minutes, a precipitate formed. The mixture was allowed to stand at room temperature for 4 hours, then the solid precipitate was collected on a Buchner funnel, washed with absolute EtOH and dried under vacuum at 85 ° C for 24 hours to give the title compound (94%) as

-4848 pale yellow crystals, m.p. 234-236 ° C. Analysis calculated for C17H24N4O2S2O, 5-Tartaric acid: C, 50.09; H, 5.97; N, 12.29; S, 14.07. Found: C, 49.85; H, 5.90; N, 12.12; S, 13.75.

Example 13: N - [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide, 0,5-D-tartrate

Warm solutions of N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide (1.00 g, 2.63 mmol) in absolute EtOH ( 40 ml) was added a solution of D-tartaric acid (198 mg, 1.32 mmol, 0.5 eq) in absolute EtOH (4 ml). After a few minutes, a precipitate formed. The mixture was allowed to stand at room temperature for 18 hours, then the solid precipitate was collected on a Buchner funnel, washed with absolute EtOH and dried under vacuum at 65 ° C for 6 hours to give the title compound (73%) as a white solid, m.p. . Mp 232-233 ° C. Analysis calculated for C17H24N4O2S2O, 5-D-tartaric acid: C, 50.09; H, 5.97; N, 12.29; S, 14.07. Found: C, 49.75; H, 5.81; N, 12.04; S, 13.37.

Example 14: N - [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide, 0,5-fumarate

Warm solutions of N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl-4-piperidinecarboxamide (1.75 g, 4.6 mmol) in absolute EtOH (100 a solution of fumaric acid (276 mg, 2.3 mmol, 0.5 eq) in absolute EtOH (5 ml) was added. After 10 minutes, a precipitate began to form. The mixture was allowed to stand at room temperature for 2 hours

-4949 and then at 5 ° C for 16 hours. The resulting solid precipitate was collected on a Buchner funnel, washed with absolute EtOH and dried under vacuum at 65 ° C for 24 hours to give the title compound (84%) as a white solid, m.p. Mp 206-207 ° C. Analysis calculated for C17H24N4O2S20.5-fumaric acid: C, 52.04; H, 5.98; N, 12.77; S, 14.62. Found: C, 51.74; H, 5.76; N, 12.57; S, 14.19. Recrystallization (95% aq. EtOH) gave the title compound containing 1 mole of EtOH (83%) as large colorless crystals, m.p. Mp 212-214 ° C. Analysis calculated for C17H24N4O2S2O, 5-fumaric acid-EtOH: C, 52.05; H, 6.66; N, 11.56; S, 13.23. Found; C, 52.03; H, 6.06; N, 11.50; S, 12.99.

Example 15: N- [5 - [[[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide, Q, 5-succinate

Warm solutions of N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide (50 mg, 0.13 mmol) in absolute EtOH (2 ml ) was added a solution of succinic acid (7.7 mg, 0.065 mmol, 0.5 eq) in absolute EtOH (0.25 ml). After 10 minutes, a precipitate began to form. The mixture was allowed to stand at room temperature for 1 hour, then the precipitate was collected on a Buchner funnel, washed with absolute EtOH and dried under vacuum at 100 ° C for 24 hours to give the title compound (70%) as a white solid, m.p. 190-192 ° C. Analysis calculated for C17H24N4O2S2O, 5-succinic acid-0.46H ₂ O: C, 50.96; H, 6.28; N, 12.51; S, 14.32. Found: C, 50.96; H, 6.20; N, 12.49; S, 14.23.

Example 16: N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide, salt of 0,5-sulfuric acid

-5050

Warm solutions of N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide (50 mg, 0.13 mmol) in absolute EtOH (2 ml ) we added 1M conduit. sulfuric acid solution (0.065 ml, 0.065 mmol, 0.5 eq). Almost immediately the precipitate formed. The mixture was cooled to 5 ° C for 2 hours, then the precipitate was collected on a Buchner funnel, washed with absolute EtOH and dried under vacuum at 100 ° C for 24 hours to give the title compound (79%) as a white solid, m.p. 256-258 ° C. Analysis calculated for C17H24N4O2S20.5H ₂ SO ₄ O, 68H ₂ O: C, 46.22; H, 6.01; N, 12.68; S, 18.14. Found: C, 46.21; H, 5.95; N, 12.71; S, 18.23.

Example 17: N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide, 0.5-citrate

Warm solutions of N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide (50 mg, 0.13 mmol) in absolute EtOH (2 ml ) was added a solution of citric acid (8.3 mg, 0.043 mmol, 0.33 eq). The solution was cooled to 5 ° C for 18 hours, then a precipitate formed, which was collected on a Buchner funnel, washed with absolute EtOH and dried under vacuum at 100 ° C for 24 hours to give the title compound (68%) as white solid, m.p. 214-216 ° C. Analysis calculated for C17H24N4O2S2 0.5-citric acid-0.10H ₂ O: C, 50.21; H, 5.94; N, 11.71; S, 13.40. Found; C, 50.21; H, 6.01; N, 11.83; S, 13.44.

Example 18: N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-5151

Suspensions of N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide (100 mg, 0.26 mmol) in isopropyl alcohol (0.75 methanesulfonic acid (0.017 ml, 0.26 mmol, 1 eq) was added. The suspension was heated to 70 ° C to give a clear solution and then methyl t-butyl ether (1.5 ml) was added. Within 15 minutes, a precipitate formed. The resulting mixture was stirred at 55 ° C for 2 hours, then at room temperature for 14 hours. The resulting precipitate was collected by filtration, then dried under vacuum at 50 ° C for 14 hours to give the title compound (85%) as a colorless powder, m.p. 105 ° C. Analysis calculated for C17H24N4O2S2MSKH ₂ O: C, 43.70; H, 6.11; N, 11.32; S, 19.44. Found: C, 43.53; H, 6.14; N, 11.15; S, 19.15.

Example 19: N - [5 - [[[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide, salt of 0,5-D, L-iabolic acid

Solutions of N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide (100 mg, 0.26 mmol) in isopropyl alcohol (0 , 80 ml) at 70 ° C was slowly added a solution of D, L-malic acid (35 mg, 0.13 mmol, 0.5 eq) in isopropyl alcohol (0.3 ml). A precipitate formed immediately. The resulting mixture was stirred at 55 ° C for 2 hours, then at room temperature for 14 hours. The precipitate was collected by filtration, then dried under vacuum at 50 ° C for 14 hours to give the title compound (75%) as a colorless powder, m.p. 216 ° C. Analysis calculated for C17H24N402S2 0.5-C4H605 H ₂ O: C, 50.98; H, 6.08; N, 12.51; S, 14.32. Found: C, 50.55; H, 6.17; N, 12.29; S, 14.05.

Claims (59)

PATENT APPLICATIONS A compound of formula I (D and its enantiomers, diastereomers, solvates and pharmaceutically acceptable salts, wherein R is alkyl; R ^{1 is} hydrogen or alkyl; X NR ² or CHNR ² R ³ ; R ² and R ³ are each independently hydrogen, alkyl, substituted alkyl, cycloalkyl or substituted cycloalkyl; and n is 0, 1, 2 or 3. A compound according to claim 1, wherein: R is alkyl; R ^{1 is} hydrogen; X NR ² or CHNR ² R ³ ; R ² and R ³ are each independently hydrogen, alkyl, substituted alkyl, cycloalkyl or substituted cycloalkyl; and n is 2. The compound of claim 1 of formula Ia -5353 and its enantiomers, diastereomers, solvates and pharmaceutically acceptable salts, wherein R ^{2 is} hydrogen, alkyl, substituted alkyl or cycloalkyl. The compound of claim 1 of formula Ib and its enantiomers, diastereomers, solvates and pharmaceutically acceptable salts, wherein R ^{2 is} hydrogen, alkyl, substituted alkyl or cycloalkyl. The compound of claim 1 of formula le and its enantiomers, diastereomers, solvates and pharmaceutically acceptable salts, wherein R ² and R ^{3 are} independently hydrogen, alkyl, substituted alkyl or cycloalkyl. A compound according to claim 1 selected from the group consisting of: N - [5 - [[[5- (1,1-dimethylethyl) -oxazolylmethylthio] -thiazolyl] piperidinecarboxamide; (±) -N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -3-piperidinecarboxamide; (±) -1- (2,3-dihydroxypropyl) - N - [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] 2-thiazolyl] -4-piperidinecarboxamide; -5454 N - [5 - [[[5- (1,1-dimethylethyl) -2-oxazolylmethyl] thio] -2-thiazolyl] -1- (1-methylethyl) -4piperidinecarboxamide; 1-cyclopropyl-N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide; N - [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -1- (2hydroxyethyl) -4-piperidinecarboxamide; (R) -N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolylmethyl] thio] -2-thiazolyl] -3-piperidinecarboxamide; (S) - N - [5 - [[(5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -3-piperidinecarboxamide; cis-4-amino-N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolylcyclohexylcarboxamide; and trans-4-amino- N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] cyclohexylcarboxamide; and its enantiomers, diastereomers, solvates and pharmaceutically acceptable salts. 7. N- [5 - [[[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -4-piperidinecarboxamide and its enantiomers, diastereomers, solvates and pharmaceutically acceptable salts. 8. (±) - N - [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -3-piperidinecarboxamide and its enantiomers, diastereomers, solvates and pharmaceutically acceptable salts. 9. (R) - N - [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -3-piperidinecarboxamide and its pharmaceutically acceptable salts. 10. (S) -N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolyl] -3-piperidinecarboxamide and its pharmaceutically acceptable salts. 11. cis-4-Amino-N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolylcyclohexylcarboxamide and its pharmaceutically acceptable salts. 12. Trans-4-amino-N- [5 - [[[5- (1,1-dimethylethyl) -2-oxazolyl] methyl] thio] -2-thiazolylcyclohexylcarboxamide and its pharmaceutically acceptable salts. -5555 A pharmaceutical composition comprising a compound of claim 1 and a pharmaceutically acceptable carrier. A pharmaceutical composition comprising a compound of claim 1 in combination with a pharmaceutically acceptable carrier and an anticancer agent formulated at a given dose. A pharmaceutical composition comprising a compound of claim 1 in combination with a pharmaceutically acceptable carrier and a p53 transactivation modulator formulated at a given dose. Use of an effective amount of an apoptosis modulating compound according to claim 1 for the manufacture of a medicament for modulating apoptosis in mammals in need thereof. Use of an effective amount of a protein kinase inhibiting compound of claim 1 for the manufacture of a medicament for inhibiting protein kinases in mammals in need thereof. Use of an effective amount of a cyclin-dependent kinase inhibiting compound according to claim 1 for the manufacture of a medicament for inhibiting cyclin-dependent kinases in mammals in need thereof. Use of an effective amount of a cdc2 inhibiting compound of claim 1 for the manufacture of a medicament for inhibiting cdc2 (cdk1) in mammals in need thereof. Use of an effective amount of a cdk2 inhibiting compound according to claim 1 for the manufacture of a medicament for inhibiting cdk2 in mammals in need thereof. Use of an effective amount of a cdk3 inhibiting compound according to claim 1 for the manufacture of a medicament for inhibiting cdk3 in mammals in need thereof. -5656 Use of an effective amount of a cdk4 inhibitor compound according to claim 1 for the manufacture of a medicament for cdk4 inhibition in mammals in need thereof. Use of an effective amount of a cdk5 inhibitor compound according to claim 1 for the manufacture of a medicament for inhibiting cdk5 in mammals in need thereof. Use of an effective amount of a cdk6 inhibitory compound according to claim 1 for the manufacture of a medicament for inhibiting cdk6 in mammals in need thereof. Use of an effective amount of a cdk7 inhibitor compound according to claim 1 for the manufacture of a medicament for inhibiting cdk7 in mammals in need thereof. Use of an effective amount of a cdk8 inhibiting compound according to claim 1 for the manufacture of a medicament for inhibiting cdk8 in mammals in need thereof. Use of a therapeutically effective amount of a composition of claim 13 for the manufacture of a medicament for the treatment of laundering iterative diseases in mammals in need thereof. Use of a therapeutically effective amount of a composition according to claim 13 for the manufacture of a medicament for the treatment of cancer in a mammal in need thereof. Use of a therapeutically effective amount of a composition according to claim 13 for the manufacture of a medicament for the treatment of inflammation, inflammatory bowel disease or graft rejection in mammals in need thereof. Use of a therapeutically effective amount of a composition according to claim 13 for the manufacture of a medicament for the treatment of arthritis in mammals in need thereof. Use of a therapeutically effective amount of a composition according to claim 14 for the manufacture of a medicament for the treatment of proliferative diseases in mammals in need thereof. Use of a therapeutically effective amount of a composition according to claim 14 for the manufacture of a medicament for the treatment of cancer in a mammal in need thereof. -5757 Use of a therapeutically effective amount of a composition according to claim 15 for the manufacture of a medicament for the treatment of proliferative diseases in mammals in need thereof. Use of a therapeutically effective amount of a composition according to claim 15 for the manufacture of a medicament for the treatment of cancer in a mammal in need thereof. Use of an effective amount of at least one of the compounds of claim 1 for the manufacture of a medicament for the treatment of cyclin-dependent kinase-related abnormalities in mammals in need of such treatment. Use of a therapeutically effective amount of a compound of claim 1 for the manufacture of a medicament for the treatment of chemotherapy-induced alopecia, chemotherapy-induced thrombocytopenia, chemotherapy-induced leukopenia or mucocitis in mammals in need of such treatment. The compound of claim 1, wherein said pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maieate, citrate, citrate and citrate The compound of claim 2, wherein said pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maieate, citrate, citrate and citrate The compound of claim 3, wherein said pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maieate, citrate, citrate and citrate The compound of claim 4, wherein said pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, -5858 Mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maleate, citrate, methanesulfonate, bromate and iodate salts. The compound of claim 5, wherein said pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maleate, citrate, methane, citrate, nitrate. The compound of claim 6, wherein said pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maleate, citrate, citrate, citrate, The compound of claim 7, wherein said pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maleate, citrate, citrate, citrate, The compound of claim 8, wherein said pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maleate, citrate, citrate, citrate The compound of claim 9, wherein said pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maleate, citrate, citrate, citrate, The compound of claim 10, wherein said pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maleate, citrate, citrate, citrate. -5959 The compound of claim 11, wherein said pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maleate, citrate, citrate, citrate, The compound of claim 12, wherein said pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maleate, citrate, citrate, citrate, The pharmaceutical composition of claim 13, wherein said pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maleate sulphate, citrate, citrate . The pharmaceutical composition of claim 14, wherein said pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maleate sulphate, citrate, citrate . The pharmaceutical composition of claim 15, wherein said pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maleate sulphate, citrate, citrate . The use of claim 17, wherein said pharmaceutically acceptable salt of said compound is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maleate sulphonate citrate sol. -6060 The use of claim 18, wherein said pharmaceutically acceptable salt of said compound is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maleate, citrate, citrate iodate salt. The use of claim 20, wherein said pharmaceutically acceptable salt of said compound is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maleate, citrate, citrate sol. The use of claim 27, wherein said pharmaceutically acceptable salt of said compound is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maleate, citrate, citrate sol. The use of claim 28, wherein said pharmaceutically acceptable salt of said compound is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maleate, citrate, citrate sol. The use of claim 31, wherein said pharmaceutically acceptable salt of said compound is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maleate, citrate, citrate sol. The use of claim 32, wherein said pharmaceutically acceptable salt of said compound is selected from the group consisting of hydrochloride, -6161 Dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maleate, citrate, methanesulfonate, bromate and iodate salts. The use of claim 36, wherein said pharmaceutically acceptable salt of said compound is selected from the group consisting of hydrochloride, dihydrochloride, sulfate, trifluoroacetate, a mixture of trifluoroacetate and hydrochloride, tartrate, fumarate, succinate, maleate, citrate, citrate sol.