CZ9901630A3 - Amines of N-(aryl/heteroaryl/alkylacetyl)amino acids, pharmaceutical compositions in which they are comprised and inhibition methods of releasing beta-amyloid peptide and/or synthesis thereof by making use of such compounds - Google Patents

Description

Amines of N- (aryl / heteroaryl / alkylacetyl) amino acids, pharmaceutical compositions containing them, and methods of inhibiting β-amyloid peptide release and / or synthesis using such compounds

Cross-references to related applications

This application relates to the following U.S. Pat. Preliminary applications:

1. U.S. Pat. Provisional Application No 60 / formed on the basis of Clause 37 of C.F.R. 1.53 (b) (2) (ii) of U.S. Pat. Patent Applications No. 08 / 754,895, filed Nov. 22, 1996; and

2. U.S. Pat. Provisional Application No 60 / formed on the basis of Clause 37 of C.F.R. 1.53 (b) (2) (ii) of U.S. Pat. Patent Applications No. 08 / 807,538, filed Feb. 28, 1997;

which are incorporated by reference in their entirety.

Technical field

The present invention relates to compounds that inhibit the release of β-amyloid peptide and / or its synthesis and are therefore useful in the treatment of Alzheimer's disease. The present invention also relates to pharmaceutical compositions comprising such compounds as well as methods of inhibiting the release of β-amyloid peptide.

Reference

The following publications, patents and patent applications are cited in this application under the numbers indicated as superscripts:

¹ Glenner et al. Alzheimer's Disease: Initial Report on Purification and Characterization of Novel Cerebrovascular Amyloid Protein, Biochem. Biophys, Res. Commun., 120: 885-890 (1984).

² Glenner et al. Polypeptide Marker for Alzheimer's Disease and Its Use for Diagnosis No. 4,666,829 issued May 19, 1987.

³ Selkoe, " The Molecular Pathology of Alzheimer's Disease, Neuron, 6: 481-498 (1991).

⁴ Goate et al., Segregation of a Missense Mutation in the Amyloid Precursor Protein Gene with Familial Alzheimer's Disease, Nature, 349: 704-106 (1990).

⁵ Chartier-Harlan et al., Early-Onset Alzheimer's Disease Caused by Mutations at Codon 711 of the β-Amyloid Precursor Protein Gene, Nature, 353: 844-846 (1989).

⁶ Murrell et al., A Mutation in the Amyloid Precursor Protein Associated with Hereditary Alzheimer's Disease, Science, 254: 97-99 (1991).

⁷ Mullan et al., A Pathogenic Mutation for Probable Alzheimer's Disease in the APP Gene at the Term-Terminus of βAmyloid, Nature Genet., 1: 345-341 (1992).

⁸ Schenk, et al., "Methods and Compositions for the Detection of Soluble β-Amyloid Peptide International Patent ApplicationPublication No. WO 94/10569. published May 11, 1994.

⁹ Selkoe, Amyloid Protein and Alzheimer's Disease, Scientific American 2-8, November 1991.

¹⁰ Tetrahedron Letters, 34 (48), 7685 (1993)) ¹¹ Losse et al., Tetrahedron, 27: 1423-1434 (1971).

¹² Citron et al., Mutation of the β-Amyloid Precursor Protein in Familial Alzheimer's Disease Increases β-Protein Production, Nature, 360: 672-674 (1992).

¹³ Hansen et al., "Reexamination and Further Development of a Precise and Rapid Dye Method for Cell Growth Measurement / Cell Kill, J. Immun. Meth., 119: 203-210 (1989).

All of the aforementioned publications, patents and patent applications are herein incorporated by reference in their entirety in the same manner as if each individual publication, patent or patent application were specifically and individually referred to as references incorporated in their entirety.

BACKGROUND OF THE INVENTION

Alzheimer's disease (AD) is a degenerative brain disorder characterized by clinically progressive loss of memory, cognitive ability, thinking, judgment, and emotional stability, which gradually lead to profound mental damage and ultimately death. Alzheimer's disease is a very common cause of the progressive mental failure (dementia) of the elderly and is believed to be the fourth leading cause of death in the United States. Alzheimer's disease has been observed in various races and ethnic groups around the world and is one of the most serious current and future public health problems. It is estimated that the disease currently affects two to three million people in the United States alone. Alzheimer's disease is currently incurable. At present, there is no known treatment method that effectively prevents or removes Alzheimer's disease.

The brains of people suffering from Alzheimer's disease show characteristic lesions called senile (or amyloid) plaques, amyloid angiopathy (amyloid deposited in blood vessels), and neurofibrillary kinking. Patients with Alzheimer's disease generally have a large number of these lesions, particularly amyloid plaques and neurofibrillary kinks, in a number of areas of the human brain that are important for memory and cognitive functions. Fewer of these lesions with more limited anatomic distribution are also found in the brain of most elderly people who do not have clinical Alzheimer's disease. Amyloid plaques and amyloid angiopathy also characterize the brain of persons with Trisomy 21 (Down syndrome) and hereditary cerebral hemorrhage with Dutch-type amyloidosis (HCHWA-D). At present, a definitive diagnosis of Alzheimer's disease usually requires observation of the above lesions in the brain tissue of patients who have died of the disease or, rarely, in small biopsy specimens of brain tissue collected during an invasive neurosurgical procedure.

The basic chemical constituent of the amyloid plaques and vascular amyloid deposits (amyloid angiopathy) characteristic of Alzheimer's disease and other disorders mentioned above is a protein having molecular weight of approximately 4.2 kilodaltons (kD) _of comprising about 39 to 43 amino acids, which is designated as β-amyloid peptide (βΑΡ) i or sometimes Αβ, ΑβΡ or β / Α4. The β-amyloid peptide was first purified and its partial amino acid sequence was determined by Glenner et al. ¹ A method for isolation and sequence data of the first 28 amino acids are described in U.S. Patent Nos. 4,666, 829 ^second

Molecular biological and protein-chemical analyzes have shown that β-amyloid peptide is a small fragment of a much larger precursor protein (APP) that is normally produced by cells in many tissues of various animals, including humans. Knowledge of the structure of the APP gene code has shown that the β-amyloid peptide is formed as a peptide segment that is cleaved from APP by the protease enzyme or enzymes. The exact biochemical mechanism by which the βamyloid peptide fragment is cleaved from APP and subsequently deposited as amyloid plaques in brain tissue and in the walls of cerebral and meningeal blood vessels is currently unknown.

Numerous evidence suggests that cerebral deposition of the βamyloid peptide plays a central role in the pathogenesis of Alzheimer's disease and may prevent cognitive symptoms by years or decades, see, for example, Selkoe ³ . The most important evidence is the discovery that DNA Mis-sense mutations at amino acid 717 isoform APP 770 amino acids may be found in affected members but not in unaffected members of many families with a genetically determined (familial) form of Alzheimer's disease (Goate et al. ^4; Chartier Harlan et al ⁵ and Murrell et al ⁶ ) and is referred to as the Swedish variant. A double mutation consisting of a mutation of lysine ^595- methionine ⁵⁹⁶ to asparagine ⁵⁹⁵ -leucine ⁵⁹⁶ (referring to isoform 695) and found in the Swedish family was described in 1992 (Mullan et al. ⁷ ). Genetic linkage analyzes have shown that these mutations, as well as certain other mutations in the APP gene, are a specific molecular cause of Alzheimer's disease in affected members of such families. In addition, mutations in amino acid 693 in the APP isoform of 770 amino acids have been identified as the cause of the β-amyloid peptide deposition, HCHWA-D, and the alanine to glycine alteration at amino acid 692 appears to cause a phenotype resembling Alzheimer's disease in some patients, but HCHWA-D in others. The discovery of these and other APP mutations in genetically induced cases of Alzheimer's disease demonstrates that altering APP and subsequently storing the β-amyloid peptide fragment can cause Alzheimer's disease. ^{I *}

Despite the progress made in understanding the mechanism underlying Alzheimer's disease and other β-amyloid peptide related diseases, there remains a need to develop methods and compositions for treating a disease or disease. Ideally, the methods of treatment would be based on agents capable of inhibiting β-amyloid peptide release and / or its synthesis in vivo.

Object of the invention

The present invention relates to the discovery of a class of compounds that inhibit the release of β-amyloid peptide and / or its synthesis and therefore are useful in preventing Alzheimer's disease in patients susceptible to Alzheimer's disease and / or treating patients suffering from Alzheimer's disease to prevent further deterioration. their condition. A class of compounds having said properties is defined by the general formula I, which follows:

X '

R'Z .CX NH> C ⁷

CH ' ^S ' NR ³ R ³ '

wherein R ¹ a) alkyl, cycloalkyl, a system where it is selected from alkenyl, alkaryl, alkcycloalkyl, aryl, cycloalkenyl, heteroaryl and heterocyclic heteroaryl or heterocyclic groups are optionally substituted with 1 to 3 substituents selected from alkyl , alkoxy, aryl, aryloxy, halogen, nitro, thioalkoxy and thioaryloxy;

(b) a substituted phenyl group of formula II

R ^b 'R'

wherein R represents alkylene containing from 1 to 8 carbon atoms, m is an integer equal to 0 or 1,

R ^a and R ^a 'are independently selected from hydrogen, hydroxy, fluoro and methyl;

R ^b and R ^b 'are independently selected from hydrogen, alkyl, alkoxy, aryl, aryloxy, cyano, cycloalkyl, halogen, heteroaryl, heteroaryloxy, heterocyclic, nitro, trihalomethyl, thioalkoxy, thioaryloxy, thioheteroaryloxy and -C (O) R ⁴ wherein R ⁴ is selected from the group consisting of alkyl, aryl, alkoxy and aryloxy; and

R ^c is selected from the group consisting of hydrogen, alkyl, aryl, cyano, halogen, nitro and wherein R ^b and R ^c are fused together to form a methylenedioxy ring with a phenyl ring; and when R ^b and / or R ^b 'and / or R ^c is a fluorine atom, a chlorine atom, a bromine atom and / or a nitro group, then R ^a and / or R ^a ' may also represent a chlorine atom; a ··> «• · · · · ·

(C) 1- or 2-naphthyl substituted with 8 one to four substituents, including alkyl, alkoxy, halogen, cyano, nitro, trihalomethyl, and thioalkoxy;

R ² is selected from the group consisting of hydrogen, alkyl of from 1 to 4 carbon atoms, alkylalkoxy of from 1 to 4 carbon atoms, alkylthioalkoxy of from 1 to 4 carbon atoms; and

R ³ and R ³ 'are independently selected from the group consisting of:

(a) a hydrogen atom with the proviso that both R ³ and R ³ 'cannot be hydrogen;

(b) alkyl with the proviso that when R ³ represents a hydrogen atom, the alkyl group R ³ 'has a straight carbon chain of at least 5 carbon atoms from the nitrogen atom, the chain optionally being substituted by one or more alkyl groups and with another condition that when both R ³ and R ³ 'are alkyl, the at least one alkyl group has a carbon chain of at least 5 carbon atoms, the chain optionally being substituted by one or more alkyl groups;

(c) - (R ⁷ ) n (W) _p , wherein R ⁷ represents an alkylene group, W is selected from the group consisting of:

at position 5, 6, 7 and / or selected from the group,

, - ··· · : ’···. · - . · 49 · · · «  9 ' 9 9 ' · ' . ·  · · 10 - : ftftft 100 • ftftft • ·

Ft ft ft ft ft (i)

wherein R ⁵ , R ⁵ ', R ⁹ and R ⁹ ' are independently selected from hydrogen, hydroxyl, alkyl, substituted alkyl, alkoxy, substituted alkoxy, amino, alkylamino, dialkylamino, aryl, acyl, acylamino, acyloxy , aminoacyl, cyano, cycloalkyl, halogen, carboxyl, carboxylic esters, heteroaryl and the heterocyclic system, and

R ⁶ is selected from the group consisting of hydrogen, hydroxyl, alkyl, substituted alkyl, amino, alkylamino, dialkylamino, aryl, acyl, acylamino, acyloxy, alkoxy, substituted alkoxy, aminoacyl, cyano, cycloalkyl, halogen, carboxyl, carboxylic esters, heteroaryl, heterocyclic system and wherein R ⁶ and one of R ⁵ or R ⁵ 'are fused to form a heterocyclic ring of 4 to 10 atoms having 1 to 3 heteroatoms selected from oxygen, nitrogen and a sulfur atom;

with the proviso that when n is zero, R ⁹ and R ⁹ 'represent a hydrogen atom;

(ii) heteroaryl; ai A. a ·: t t t t iii iii iii iii iii iii iii iii iii iii iii iii iii (iii) N- heterocyclic system, with the proviso that when W is an N-heterocyclic system, then n is not zero, and n is an integer equal to 0 or 1, and p is an integer equal to 1 to 3 with the proviso that if n is zero then p is 1, and

(d) -CH (<X>), CH ₂ C (O) OQ, where Q is selected from file, including alkyl, system aryl, heteroaryl and heterocyclic X 'represents an atom fluorine; hydrogen, a hydroxy group or an atom X represents an atom hydrogen, a hydroxy group or an atom fluorine or X 'and X together form an oxo group,

Z is selected from the group consisting of a covalent bond that binds R ¹ to -CX'X-, an oxygen atom, and a sulfur atom; with the proviso that if R ¹ is phenyl, R ² is methyl, X 'and X are hydrogen, Z is a group covalently bonded to R ¹ -CX'X-, m is zero, R ³ is hydrogen , R ³ 'is - (R ⁷ ) _n (W) _p , where n is zero and _p is one and W represents

then (i) R ⁵ , R ⁵ , R ⁹ , R ⁹ and R ⁶ are not all hydrogen and

(Ii) R ⁵ , R ⁵ ', R ⁹ , R ⁹ ' are hydrogen and R ⁶ is methoxy;

with the further proviso that when R ¹ is 3,5-difluorophenyl, R ² is methyl, X 'and X are hydrogen, Z is a group that covalently binds R ¹ to C X'X "-, m is zero, R ³ is a hydrogen atom, R ³ 'is - (R ⁷ ) n (W) _p , wherein n is equal to one and _p is equal to one, R ⁷ represents ethylene and W represents

R ⁹ R ^p

R ⁹ 'R ^s' and R ^5, R ^5', R ⁹ ', R ^9' and R ⁶ do not represent all hydrogen; and with the further proviso that when R ¹ is 3,5-difluorophenyl, R ² is methyl, X 'and X are hydrogen, Z is a group that covalently binds R ¹ to C X'X "- and m is zero, R ³ is an atom hydrogen, R ³ 'is - (R ⁷ ) _n (W) p, where n is zero and p is one, W represents

> «· ► ·

R 'R ⁵

and R ⁵ ', R ⁹ , R ⁹ ' are hydrogen, then R ⁵ and R ⁶ are not condensed to form a phthalimide group with the phenyl ring to which they are attached.

Accordingly, one method of the present invention relates to a method of inhibiting the release of a βamyloid peptide and / or its synthesis in a cell, wherein an amount of a compound or mixture of compounds of formula I above effective to inhibit the cell releasing and / or synthesizing a βamyloid peptide.

Since the in vivo generation of β-amyloid peptide is associated with the pathogenesis of Alzheimer's disease ^8.9 , the compounds of Formula I can also be used together with a pharmaceutical composition for the prophylactic and / or therapeutic prevention and / or treatment of Alzheimer's disease. Accordingly, in a further aspect, the present invention relates to a prophylactic method of preventing Alzheimer's disease in a patient at risk of developing Alzheimer's disease, said method comprising administering a pharmaceutical composition comprising a pharmaceutically inert carrier and an effective therapeutic agent. · 0 0 • 0 · 000 · 00

an amount of a compound or mixture of compounds of formula I above to said patient.

In another aspect, the present invention relates to a therapeutic method of treating a patient suffering from Alzheimer's disease for inhibiting further deterioration of said patient, said method comprising administering to said patient a pharmaceutical composition comprising a pharmaceutically inert carrier and an effective amount of a compound or mixture of compounds of formula I above. .

In formula I above, preferred R ¹ unsubstituted aryl groups include, for example, phenyl,

1-naphthyl, 2-naphthyl and the like.

Preferred R ¹ substituted aryl groups include, for example, monosubstituted phenyls having a single substitution at the 2, 3 or 4 position, wherein each of the individual substituents is given by the corresponding groups R ^a , R ^b and R ^c ; disubstituted phenyls that include phenyls having two substituents at 2,3-positions, 2,4-positions, 2,5-positions, 2,6-positions, 3,4-positions, 3,5-positions, or v-positions positions 3,6 wherein each of the substituents is given by the corresponding groups R ^a , R ^a ', R ^b , R ^b ' and R ^c ; and trisubstituted phenyls which include phenyls having three substituents at positions 2,3,4, at positions 2,3,5, at positions 2,3,6, at positions 3,4,5 and at positions 3,4, 6, wherein each of the substituents is given by the corresponding groups R ^a , R ^a ', R ^b , R ^b ' and R ^c . Preferably, substituted phenyl groups do not include more than 3 substituents.

9 «· 9 9 9 99 «9 9 9 9 9 9 9 9 99 9 9 · 9 9 9 999 9 9 9 9 9 9 9 9 9 9 9 9 9 e

Examples of substituted phenyls include, for example, 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, 4-nitrophenyl, 4-methylphenyl, 3-methoxyphenyl, 3-nitrophenyl, 3-fluorophenyl, 3-chlorophenyl, 3-bromophenyl, 3-thiomethoxyphenyl, 3-methylphenyl, 3-trifluoromethylphenyl , 2-hydroxyphenyl, 2-methylphenyl, 2-fluorophenyl, 3,4-dichlorophenyl, 3,4-methylenedioxyphenyl,

3,5-difluorophenyl, 3,5-dichlorophenyl, 2,4-dichlorophenyl and 2,5-difluorophenyl.

Preferred R ¹ alkaryl groups include, by way of example, benzyl, 3-phenylethyl, 4-phenyl-n-propyl and the like.

Preferred R ¹ alkyl, cycloalkyl and cycloalkenyl groups include, by way of example, sec-butyl, cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, cyclohexl-enyl, -CH ₂ -cyclopropyl, -CH ₂ -cyclobutyl, -CH ₂ -cyclohexyl, -CH ₂ -cyclopentyl, -CH ₂ CH ₂ -cyclopropyl, -CH ₂ CH ₂ -cyclobutyl, CH ₂ CH ₂ -cyclohexyl, -CH ₂ CH ₂ -cyclopentyl and the like.

Preferred R ¹ heteroaryls and substituted heteroaryls include, by way of example, pyrid-3-yl, pyrid-4-yl, thiophen-2-yl, thiophen-3-yl, benzothiazol-4-yl, 2-phenylbenzoxazol-5-yl, furan -2-yl, benzofuran-2-yl, thionaphthen-2-yl, 2-chlorothiophen-5-yl, 3-methylisoxazol-5-yl, 2- (thiophenyl) thiophen-5-yl, 6-methoxythionaphthen-2-yl, 3-phenyl-1,2,4-thiooxadiazol-5-yl, 2-phenyloxazol-4-yl and the like.

Preferably, R ² is selected from the group consisting of alkyl of from 1 to 4 carbon atoms, alkylalkoxy of from 1 to 4 carbon atoms, and alkylthioalkoxy of from 1 to 4 carbon atoms. do do do do do do do do do do do do ♦ φ φ do φ do do do do do do do do do do do do do do do do do do do do do do do do do do do do do do do do do do Particularly preferred R ² substituents include, by way of example, methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, -CH ₂ CH ₂ SCH _3, and the like.

Preferably, R ³ represents a hydrogen atom and R ³ 'is selected from the group consisting of 3-hydroxyphenyl, 3-methoxyphenyl, 3-methoxycarbonylphenyl, n-hexyl, n-octyl, 4-methoxycarbonylphenyl, 4-methoxycarbonylphenyl, 3-chlorophenyl, 3-cyanophenyl, 3, 5-dichlorophenyl, -CH _(CH3) <I> (R stereoisomer), -CH (CH ₃₎ O (S stereoisomer), phthalid-6-yl, 2-hydroxypyrid-3-yl, 2- (methoxycarbonylmethyl) benzyl, 3 (methoxycarbonyl) benzyl, 2- (2'-methoxycarbonylmethylphenyl) benzyl and 2-phenylbenzyl.

Particularly preferred compounds for use in the methods and compositions of the present invention include, by way of example, the following compounds:

N- (3-hydroxyphenyl) -N '- (phenylacetyl) -L-alaninamide

N- (3-methoxyphenyl) -1 '- (phenylacetyl) -L-alaninamide

N- (3-ethoxyphenyl) -1 '- (phenylacetyl) -L-alaninamide

N- (4-ethoxycarbonylphenyl) -1 '- (phenylacetyl) -L-alaninamide

N- (n-hexyl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide

N- (n-octyl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide

N- (3-methoxyphenyl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide • φφ «· • * · * * * · ΦΦΦΦ · φ φ φ · · · · · ·

· 9 9

ΦΦΦ · Φ ΦΦΦ ΦΦΦ ΦΦ ΦΦ

N- (4-ethoxycarbonylphenyl) -1 '- (3,5-difluorophenylacetyl) -alaninamide

N- (3-ethoxycarbonylphenyl) -1 '- (3,5-difluorophenylacetyl) -Lalaninamide

N- (3-chlorophenyl) -N '- (3,5-difluorophenylacetyl) -L-alaninamide

N- (3,5-dichlorophenyl) -1 '- (3,5-difluorophenylacetyl) -alaninamide

N- (3-cyanophenyl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide

N- (phthalid-6-yl) -N '- (3,5-difluorophenylacetyl) -L-alaninamide

N - [(4-methoxycarbonylphenyl) methyl] -N '- (3,5-difluorophenylacetyl) -L-alaninamide

N- (1-cyano-1-phenylmethyl) -1 '- (3,5-difluorophenylacetyl) -Lalaninamide

N - [(R) -1-phenylethyl] -N '- (3,5-difluorophenylacetyl) -Lalaninamide

N - [(S) -1-Phenylethyl] -N '- (3,5-difluorophenylacetyl) -Lalaninamide

N- [2-hydroxypyridin-3-yl] -1 '- (3,5-difluorophenylacetyl) -Lalaninamide

0 »• 0 0 ·

0 0 0000

0

0 0 0 0 0 0

N- [2-methoxycarbonyl-1-phenylethyl] -1 '- (3,5-difluorophenylacetyl) -L-alaninamide

N - [α-Pyridin-2-yl-benzyl] -1 '- (3,5-difluorophenylacetyl) -Lalaninamide

N- [1- (N-phthalamido) pent-2-yl] -1 '- (3,5-difluorophenylacetyl) -Lalaninamide

N- [2- (methoxycarbonylmethyl) benzyl] -1 '- (3,5-difluorophenylacetyl) -L-alaninamide

N- [3- (methoxycarbonyl) benzyl] -N '- (3,5-difluorophenylacetyl) L-alaninamide

N- [2- (2'-Methoxycarbonylmethylphenyl) benzyl] -1 '- (3,5-difluorophenylacetyl) -L-alaninamide

N- [2-phenylbenzyl] -N '- (3,5-difluorophenylacetyl) -L-alaninamide ^: 4 ^: 4 ^: 4 ^: 4 ^: 4 ^: 4. 4 · 4 44 ··· “4 4 · · 4

Still further, the present invention relates to the novel compounds of formula I above. Preferred compounds have the general formula II shown in Table I, which follows:

.CHR ¹

II

O

CH

NHR ³

R ¹ R ² R ³ ' -Φ -ch ₃ 3-HO-6- -Φ -ch ₃ 3-CH ₃ O - -Φ -ch ₃ 3-CH ₃ CH ₂ O (CO) -O- -Φ -ch ₃ 4-CH ₃ CH ₂ O (O) C-6- 3,5-di-F - -ch ₃ CH ₃ (CH ₂ ) _s -

·· • ·

9 9 9

999 999 ·

1 R · R ² R ³ ' 3,5-di-F- <£ - -ch ₃ CH, (CH ₂ ) ₇ - 3,5-di-F - -ch. 3-CH, 0 - 3,5-di-F- <-> - -ch ₃ 4-CH ₃ CH ₂ O (O) C-E- 3,5-di-F -ch ₃ 3-CH, CH ₂ O (O) C - 3,5-di-F - -CH ₃ 3-Cl - 3,5-di-F - -ch ₃ 3,5-di-Cl-et- 3,5-di-F - -ch ₃ 3-Cyano - </> - 3,5-di-F - -ch. phthalid-6-yl 3,5-di-F - -CH, 4-CH ₃ O (O) C-2-CH ₂ - 3,5-di-F - -CH, a-cyanobenzyl- 3,5-di-F - -ch ₃ (R) -CH ₃ (s) CH- 3,5-di-F - -Ch, (S) -CH, (</>) CH- 3,5-di-F - ^ - -CH, 2-Hydroxypyridin-3-yl 3,5-di-F- - -CH, α-pyridin-2-yl-benzyl | 3,5-di-F - -CH,. 1- (N-phthalamido) pent-2-yl I 3,5-di-F - -CH, 2- (methoxy- carbonylmethyl) benzyl | 3,5-di-F-tf> - -CH, 3- (methoxy- carbonyl) benzyl | 3,5-di-F - -CH, 2- (2'-methoxy- carbonylmethyl- fenyDbcnzyl 3,5-di-F - -CH ₃ 2-phenylbenzyl | 3,5-di-F - -ch ₃ 2-methoxycarbonyl-1-phenylmethyl |

99 9 99 9 9 9 · • 99 9 9 9 9 9 9 99 9 9 9 9 9 9 «· 9 9 9 9 9 99 9 99 9 • · 9 • 9 9 9 * 9 9 9 • 9 · 999 9 9 9 9

DETAILED DESCRIPTION OF THE INVENTION

As mentioned above, the present invention relates to compounds that inhibit the release of β-amyloid peptide and / or its synthesis and accordingly can be used in the treatment of Alzheimer's disease. Before the present invention is described in further detail, the following terms will first be defined.

Definition

The term β-amyloid peptide refers to a peptide of 39-43 amino acids having a molecular weight of about 4.2 kD, the peptide being substantially homologous to the form of protein described by Glenner et al. ¹ , including mutations and post-translational modifications of the normal βamyloid peptide. In whatever form, the βamyloid peptide is a fragment of approximately 39-43 amino acids of the larger transmembrane glycoprotein, referred to as the β-amyloid precursor protein (APP). Its 43-amino acid sequence is as follows:

1 Asp Ala Glu Phe Arg His Asp Ser Gly Tyr 11 Glu Wall His His Gin Lys Leu Wall Phe Phe 21 Ala Glu Asp Wall Gly Ser Asn Lys Gly Ala 31 Ile Ile Gly Leu Met Wall Gly Gly Wall Wall 41 Ile Ala Thr (SEQ ID NO: 1)

or a sequence that is

• 4 4 • 4 · 4 4 4 is essentially homologous.

Alkyl refers to monovalent alkyl groups that preferably contain from 1 to 20 carbon atoms, preferably 1 to 10 carbon atoms, and more preferably 1 to 6 carbon atoms. Examples include methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, nhexyl and the like.

Substituted alkyl refers to an alkyl group, preferably containing from 1 to 10 carbon atoms, having from 1 to 3 substituents selected from the group consisting of alkoxy, substituted alkoxy, acyl, acyloxy, acylamino, amino, aminoacyl, aminocarboxy esters, cyano, cycloalkyl, halogen, hydroxyl, carboxyl, carboxylic esters, thiol, thioalkoxy, substituted thioalkoxy, aryl, heteroaryl, heterocyclic, nitro and mono- and di-alkylamino, mono- and di (substituted alkyl) amino, mono- and di-arylamino, mono- and di-heteroarylamino, mono- and di-heterocyclic amino groups and unsymmetrical di-substituted amines having various substituents selected from the group consisting of alkyl, substituted alkyl, aryl, heteroaryl and the heterocyclic system.

Alkylene refers to divalent alkylene groups which preferably contain from 1 to 10 carbon atoms, and more preferably 1 to 6 carbon atoms, which may be straight chain or branched. Examples include methylene (-CH ₂ -), ethylene (-CH ₂ CH ₂ -), BB. · 9 Β .. ΒΒ> - '·· ββ · ·. - ^ζ ·· ·· · «· • · β β β β β β β • Β · _··; · · · Β ··· ΒΒΒ

Β Β ·. ΒΒ · ·

Propylene isomers (e.g., -CH ₂ CH ₂ CH ₂ - and -CH (CH ₃ ) CH ₂ -) and the like.

Alkaryl refers to -alkylene-aryl groups which preferably contain from 1 to 10 carbon atoms in the alkylene moiety and from 6 to 14 carbon atoms in the aryl moiety. Such alkaryl groups are, for example, benzyl, phenethyl and the like.

Alkcycloalkyl refers to -alkylene-cycloalkyl groups which preferably contain from 1 to 10 carbon atoms in the alkylene moiety and from 3 to 8 carbon atoms in the cycloalkyl moiety. Such alkcycloalkyl groups are, for example, -CH ₂ cyclopropyl, -CH ₂ CH ₂ -cyclohexyl and the like.

Alkoxy refers to an alkyl-O- group. Preferred alkoxy groups include, by way of example, methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy and the like.

A substituted alkoxy group refers to a substituted alkyl-O- group wherein the substituted alkyl has the same meaning as previously defined.

Alkylalkoxy refers to the group -alkylene-O-alkyl, which includes, by way of example, methylenemethoxy (-CH ₂ OCH ₃ ), ethylenemethoxy (-CH ₂ CH ₂ OCH ₃ ), methylene-iso-propoxy (-CH ₂ - O-CH (CH ₃ ) ₂ ) and the like.

Alkylthioalkoxy refers to the group -alkylene-Salkyl, which includes, by way of example, methylene thiomethoxy (-CH ₂ SCH ₃ ), ethylene thiomethoxy (24 • 0 4 9 49 00)

4 98 99 0 0 0 0

4 4 0 0 0 · 9 · 999 999

9 4 4 1 5

CH2CH2SCH3), methylene-iso-thiopropoxy (-CH2SCH (CH3) 2) and the like.

Alkenyl refers to alkenyl groups which preferably contain from 2 to 10 carbon atoms and more preferably 2 to 6 carbon atoms and have at least 1 and preferably 1-2 alkenyl unsaturation sites. Preferred alkenyl groups include ethenyl (CH = CH ₂ ), n-propenyl (-CH ₂ CH = CH ₂ ), iso-propenyl (-C (CH ₃ ) = CH ₂ ), and the like.

Alkynyl refers to alkynyl groups which preferably contain from 2 to 10 carbon atoms and more preferably 2 to 6 carbon atoms and have at least 1 and preferably 1-2 alkynyl unsaturation sites. Preferred alkynyl groups include ethynyl (CH = CH ₂ ), propargyl (-CH ₂ CH = CH ₂ ) and the like.

Acyl refers to alkyl-C (O) -, substituted alkyl-C (O) -, cycloalkyl-C (O) aryl-C (O) -, heteroaryl-C (O) and heterocyclic-C (O) wherein alkyl, substituted alkyl, cycloalkyl, aryl and heteroaryl are as defined above.

Acylamino refers to the group -C (O) NRR wherein each R is independently hydrogen or alkyl.

Aminoacyl refers to the group -NRC (O) R, wherein each R is independently hydrogen or alkyl.

Acyloxy refers to alkyl-C (O) O-, substituted alkyl-C (O) O-, cycloalkyl-C (O) O-, aryl-C (O) O-, heteroaryl-C (O) O- and heterocycle-C (O) O-, wherein alkyl, substituted alkyl, cycloalkyl, aryl, heteroaryl and

Φ · · φ · • • • · · · · · · ·

Φ Φ 9 · φφφ ΦΦΦ

· Φ Φ «·

The heterocyclic system has the meaning defined above.

Aryl refers to an unsaturated aromatic carbocyclic group of 6 to 14 carbon atoms having a single ring (e.g. phenyl) or multiple condensed rings (e.g. naphthyl or anthryl). Preferred aryls include phenyl, naphthyl and the like.

Unless otherwise limited by the definition of a single substituent, such aryl groups may optionally be substituted with 1 to 3 substituents selected from the group consisting of hydroxy, acyl, acyloxy, alkyl, alkoxy, alkenyl, alkynyl, amino, aminoacyl, aryl, aryloxy, carboxyl, carboxylic esters, amino carboxylic esters, cyano, halogen, nitro, heteroaryl, heterocyclic, thioalkoxy, trihalomethyl and the like. Preferred substituents include alkyl, alkoxy, halogen, cyano, nitro, trihalomethyl and thioalkoxy.

Aryloxy refers to an aryl-Ο- group wherein the aryl group is as defined above, including optionally substituted aryl groups as also defined above.

Carboxyl refers to the group -C (O) OH; carboxylic esters refer to -C (O) O-alkyl, -C (O) O-substituted alkyl, -C (O) O-aryl, -C (O) O-heteroaryl, and C (O) O-heterocyclic; and aminocarboxylic esters refer to -NHC (O) OR groups where R is alkyl;

A substituted alkyl, aryl, cycloalkyl, heteroaryl or heterocyclic system is substituted.

Cycloalkyl refers to cyclic alkyl groups containing from 3 to 10 carbon atoms having a single cyclic ring or multiple fused rings, which may optionally be substituted with 1 to 3 alkyl groups. Such cycloalkyl groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, 1-methylcyclopropyl, 2-methylcyclopentyl, 2-methylcyclooctyl and the like or multiple ring structures such as adamantanyl and the like.

Cycloalkenyl refers to cyclic alkenyl groups containing from 4 to 10 carbon atoms having a single cyclic ring and at least one internal unsaturation site, which may optionally be substituted with 1 to 3 alkyl groups. Examples of suitable cycloalkenyl groups include, for example, cyclobut-2-enyl, cyclopent-3enyl, cyclooct-3-enyl and the like.

Halo or halogen refers to a fluorine atom, a chlorine atom, a bromine atom and an iodine atom, and preferably to either a fluorine atom or a chlorine atom.

Heteroaryl refers to a monovalent aromatic carbocyclic group containing from 1 to 10 carbon atoms and 1 to 4 heteroatoms selected from oxygen, nitrogen and sulfur within the ring.

• · • ·

4 »· · · · · · · · ·

Unless otherwise limited by the definition of a single substituent, such heteroaryl groups may be optionally substituted with 1 to 3 substituents selected from the group consisting of alkyl, alkoxy, aryl, aryloxy, halogen, nitro, heteroaryl, hydroxy, thioalkoxy, thioaryloxy and the like. Such heteroaryl groups may have a single ring (e.g. pyridyl or furyl) or multiple fused rings (e.g. indolizinyl or benzothienyl). Preferred heteroaryls include pyridyl, pyrrolyl and furyl.

Heteroaryloxy refers to a heteroaryl-Ο- group, wherein heteroaryl is as defined above, including optionally substituted heteroaryl groups as also defined above.

A heterocycle or heterocyclic system refers to a monovalent saturated or unsaturated group having a single ring or multiple fused rings containing from 1 to 10 carbon atoms and from 1 to 4 heteroatoms selected from the group consisting of nitrogen, sulfur or oxygen, inside circle.

Unless otherwise limited by the definition of a heterocyclic substituent, such heterocyclic groups may be optionally substituted with 1 to 3 substituents selected from alkyl, alkoxy, aryl, aryloxy, halogen, nitro, heteroaryl, thioalkoxy, thioaryloxy and the like. Such heterocyclic groups may have a single ring (for example piperidinyl or tetrahydrofuranyl) or multiple condensed rings.

- & - ·· ·· · · · · · · · · · · · · · · · · · · · · ·

Examples of heterocycles and heteroaryls include, but are not limited to, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazine, quinazine cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, phenanthroline, isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline, piperidine, piperazine, indoline, phthalimide, 1,2,3,4-tetrahydroisoquinoline, 4,5, 6,7tetrahydrobenzo [b] thiophene, thiazole, thiazolidine, thiophene, benzo [b] thiophene, morpholino, piperidinyl, pyrrolidine, tetrahydrofuranyl and the like.

Heterocyclyloxy refers to a heterocycle-O- group, wherein the heterocyclic system is as defined above, including optionally substituted heterocyclic groups, as also defined above.

Thiol refers to the -SH group.

Thioalkoxy refers to -S-alkyl groups wherein alkyl has the same meaning as previously defined.

Thio-substituted alkoxy refers to groups -S-substituted alkoxy, wherein the substituted alkoxy group is as defined above.

Thioaryloxy refers to an aryl-S- group wherein the aryl group is as defined above, including optionally substituted aryl groups as also defined above.

• 99 9 9 · 9

9 9 99 9 9 9 ·

98 9 9 9 9 9

Thioheteroaryloxy refers to a heteroaryl-S- group, wherein the heteroaryl group is as defined above, including optionally substituted aryl groups, as also defined above.

A pharmaceutically acceptable salt refers to a pharmaceutically acceptable salt of a compound of formula I, wherein the salts are derived from a number of organic and inorganic ions well known in the art and include, by way of example only, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium and alike; and when the molecule contains a basic functional group, salts of organic or inorganic acids such as hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, oxalate and the like.

Preparation of compounds

The compounds of formula (I) may be readily prepared by several divergent synthetic routes, the particular route being chosen due to the difficulty of preparing the compound, the commercial availability of the starting materials, and the like.

The first method of synthesis involves the conventional coupling of an acid derivative with a primary amine of an esterified amino acid as shown in reaction (1), which follows:

··

3CC ί • · · · · ··· ··· o

II

CX OH I

X '

HΉ OR 'I

R ² 2

0)

X '

II I

OR ² wherein R ¹ , R ² , X 'and X are as defined above and R ¹ is preferably a hydrogen atom or an alkyl group.

reagents such as those well known

Reaction (1) involves only the coupling of a suitable acid derivative (1) with a primary amine (2 amino acids) amino acid ester under conditions that result in the formation of the Nacetyl derivative (3). This reaction is conveniently performed in peptide synthesis, and the synthetic methods used therein can also be used to prepare the N-acetyl amino acid (amino acid ester) 3. For example, well known coupling agents can be used to facilitate coupling, such as carbodiimides, with concomitant use of additives such as N-hydroxysuccinimide, ftft, ftftft, ftftft, ftft. ftft · ftftft ftftft ft "ft" · hydroxybenzotriazole and the like. The reaction is conveniently carried out in an inert aprotic diluent such as dimethylformamide, dichloromethane, chloroform, acetonitrile, tetrahydrofuran and the like.

Alternatively, in a preferred embodiment of the present invention, the hydrogen halide compound (1) may be used in reaction (1) and when used, it is typically used in the presence of a suitable acid scavenging base formed during the reaction. Suitable bases include, by way of example, triethylamine, diisopropylethylamine, Nmethylmorpholine and the like.

Reaction (1) is preferably carried out at a temperature from about 0 ° C to about 60 ° C until completion of the reaction, which usually occurs after about 1 to about 24 hours. Upon completion of the reaction, the N-acetyl amino acid / amino acid ester / 3 is isolated by conventional means, including precipitation, chromatography, filtration, and the like, or alternatively in the case of an ester, hydrolyzed to the corresponding carboxylic acid without purification and / or isolation by other than conventional processing. (e.g., aqueous extraction and the like).

If an amino acid N-acetyl ester is formed, it is typically converted to the corresponding acid prior to coupling with the amine HNR ³ R ³ '. Coupling is accomplished using well known peptide coupling chemistry with well known coupling reagents such as carbodiimides with concomitant use of well known additives such as N-hydroxysuccinimide,

1-hydroxybenzotriazole and the like, which can be used to facilitate coupling. The reaction is conveniently carried out in an inert, aprotic polar diluent such as dimethylformamide, dichloromethane, chloroform, acetonitrile, and the like. tetrahydrofuran and the like. Alternatively, the ester group of compound (3) may in some cases be converted directly to an amide group by a conventional ester / amide exchange reaction well known in the art.

In reaction (1), each of the reagents (acetic acid derivative (1) and amino acid / amino acid ester (2)) is well known in the art and many are commercially available.

Alternatively, compounds of formula I may be prepared by first forming an amino acid amide and then Ν'-acetylating these esters. That is, the amine HNR ³ R ³ 'is coupled to the N'-blocked amino acid Block-NHCHR ² COOH under conventional coupling conditions to give the N'-blocked amino acid block Block-NHCHR ² C (O) NR ³ R ³ '. The blocking group is then removed under conventional conditions to give the free amine, which is then N'-acetylated as described above to give compounds of Formula I.

Upon completion of coupling and N'-acetylation (in any order), the resulting amides can be derivatized by conventional chemical techniques to provide derivatives of the synthesized compounds. For example, a reactive functional group that is blocked on either the R ² and / or the R ³ group can be unblocked and then derivatized. For example, a Boc-protected amino group on R ² (e.g., a lysine side chain) may be unblocked after synthesis and the amino group may be acylated or otherwise derivatized.

·· · ·

·····························

The compounds described above can also be prepared using polymeric supported carbodiimide peptide coupling reagents. For example, a polymer supported form of EDC has been described (Tetrahedron Letters, 34 (48), 1685 (1993)) ¹⁰ . In addition, a novel carbodiimide coupling reagent, PEPC and its corresponding polymer supported forms have been discovered which are very useful for preparing the compounds of the present invention.

Polymers suitable for use in preparing polymeric coupling reagents are either commercially available or can be prepared by methods well known to those skilled in the art of polymers. A suitable polymer must carry free side chains bearing groups that react with the terminal amine of carbodiimide. Such reactive groups include chlorine atom, bromine atom, iodine atom and methanesulfonyl. Preferably, the reactive group is a chloromethyl group. In addition, the polymer backbone must be inert to the carbodiimide and the reaction conditions under which the final polymer bound coupling reagent will be used.

Certain hydroxymethylated resins can be converted to chloromethylated resins useful for preparing coupling reagents on polymeric supports. Examples of these hydroxylated resins include 4-hydroxymethylphenylacetamidomethyl resin (Pam resin) and 4benzyloxybenzyl alcohol resin (Wang resin) supplied by Advanced Chemtech, Louisville, Kentucky, USA (see Advanced Chemtech 1993-1994 catalog, page 115). Hydroxymethyl groups of these resins

9 99 99

9 9 9 9 9 9

9

9 9 999999

9 9 9

The 999C 9C may be converted to the desired chloromethyl groups by any of the methods well known to those skilled in the art.

Preferred resins are chloromethylated styrene / divinylbenzene resins due to their easy commercial availability. As the name suggests, these resins are already chloromethylated and do not require any chemical modification before use. These resins are commercially known as Merrifield resins and are available from Aldrich Chemical Company, Milwaukee, Wisconsin, USA (see Aldrich 1994-1995 catalog, page 899). Methods for preparing PEPC and its polymer supported forms are outlined in the following scheme.

* ·

✓ ^LG

Functionalized resin where * inert polymer and LG = Cl, Br, I or OSO ₂ CH ₃

Such methods are described more fully in U.S. Pat. Patent Application No. 60 / 019,790, filed June 14, 1996, which is incorporated herein by reference in its entirety. Briefly, PEPC is prepared by first reacting ethyl isocyanate with 1- (3-aminopropyl) pyrrolidine. The resulting urea was treated with 4-toluenesulfonyl chloride to give PEPC. The polymer supported form is prepared by reacting PEPC with a suitable resin under standard conditions to provide the desired reagent.

Carboxylic acid coupling reactions using these reagents are carried out at a temperature of about ambient temperature to about 45 ° C, for a period of about 3 to 120 hours. Typically, the product can be isolated by washing the reaction mixture to be prepared with CHCl 3 and concentrating the remaining organic phase under reduced pressure. As discussed above, isolation of the product from the reaction mixture when polymer-bound reagent has been used is greatly simplified since it only requires filtration of the reaction mixture and then concentration of the filtrate under reduced pressure.

Other methods for preparing esters are given in the examples below.

In these methods of synthesis, the starting materials may contain a chiral center (for example alanine) and when a racemic starting material is used, the resulting product is a mixture of the R, S enantiomers. Alternatively, the chiral isomer of the starting material may be used, and if the reaction protocol used does not racise this starting material, a chiral product is obtained. Such reaction protocols may include inversion of the chiral center during synthesis.

Accordingly, unless otherwise stated, the products of the present invention are mixtures of the R, S enantiomers. However, if a chiral product is desired, the chiral product preferably corresponds to an L-amino acid derivative. Alternatively, chiral products can be obtained by purification methods that separate the enantiomers from the R, S mixture to give one or the other stereoisomer. Such techniques are well known in the art.

Pharmaceutical preparations

When used pharmaceutically, the compounds of Formula I are usually administered in the form of pharmaceutical compositions. The compounds may be administered by a variety of routes including oral, rectal, transdermal, subcutaneous, intravenous, intramuscular, and intranasal. These compounds are effective as injectable as well as oral compositions. Such compositions may be prepared in a manner well known in the pharmaceutical art and comprise at least one active compound.

The present invention also relates to pharmaceutical compositions comprising as active ingredient one or more of the compounds of formula I above together with a pharmaceutically acceptable carrier. In forming the compositions of the present invention, the active ingredient is usually mixed with an excipient, diluted with an excipient, or surrounded by a carrier, which may be in the form of capsules, wafers, sachets or other containers. When the excipient serves as a diluent, it may be a solid, semi-solid or liquid material that serves as a vehicle, carrier or medium for the active ingredient. Thus, the composition may be in the form of tablets, pills, powders, lozenges, sachets, wafers, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as solids or in liquid media), ointments containing, for example, up to 10% by weight compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders.

• 00 0 0 00 00 00 0 0 00 00 0 0

00 0 0 0000 00 00 0 0 0 0 0 0000 0 0 0 0 0 0 0

000 000 000 000 00 00

When preparing the formulation, it may be necessary to grind the active compound to achieve an appropriate particle size prior to combining with the other ingredients. If the active compound is substantially insoluble, it is usually ground to a particle size of less than 200 mesh. If the active compound is substantially water-soluble, the particle size is normally adjusted by grinding to a size that provides substantially uniform distribution in the formulation, for example to about 40 mesh.

Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starches, acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, sterile water, syrup, and methylcellulose. The formulations may further comprise: lubricating agents such as talc, magnesium stearate and mineral oil; wetting agents; emulsifying and suspending agents; preservatives such as methyl and propyl hydroxybenzoate; sweeteners and flavorings. The compositions of the present invention may be formulated so as to provide immediate, sustained or delayed release of the active ingredient upon administration to a patient, using procedures known in the art.

The compositions are preferably formulated as a unit dose, each dosage containing from about 5 to about 100 mg, more usually from about 10 to about 30 mg, of the active ingredient. The term "unit dose" means a physically discrete unit, suitable as a single dose for a human or other mammal, each unit containing in advance

4. ·; - ···. ' . 44 * '• 4 4

- * ·

4 44 449

9

The amount of active ingredient calculated to produce the desired therapeutic effect, together with a suitable pharmaceutical excipient. Preferably, the compound of formula (I) above is used such that it is no more than 20 weight percent of the pharmaceutical composition, preferably no more than 15 weight percent, while the remainder is a pharmaceutically inert carrier or carriers.

The active compound operates over a wide dosage range and is generally administered in a pharmaceutically effective amount. It will be appreciated, however, that the amount of the compound to be actually administered will be determined by the physician depending on the relevant circumstances, including the condition to be treated, the route of administration chosen, the compound being administered, the age, weight and response of the individual patient, severity. patient symptoms and the like.

For preparing solid compositions such as tablets, the main active ingredient is admixed with a pharmaceutical excipient to form a solid pretreated composition comprising a homogeneous mixture of the compounds of the present invention. When said preprocessed composition is said to be homogeneous, it is meant that the active ingredient is dispersed uniformly throughout the composition so that the composition can be readily subdivided into effective unit doses such as tablets, pills and capsules containing the same amount active substance. The solid pretreated composition is then subdivided into unit doses of the type described above containing, for example, from about 0.1 to about 500 mg of the active ingredient of the present invention.

materials such as polymeric acids; and

The tablets or pills of the present invention may be coated or otherwise formulated to provide a dosage form that provides the advantage of prolonged action. For example, tablets or pills may contain an inner dose of the ingredient and an outer dose of the ingredient, the outer dose forming an inner dose shell. The two components of the dosage form may be separated by an enteric layer which serves to prevent disintegration in the stomach and to allow the inner component to pass intact into the duodenum or to achieve delayed release. A variety of materials comprising a number of mixtures of polymeric acids with materials such as shellac, ketyl alcohol and cellulose acetate may be used to form such an enteric layer or coating.

Liquid forms in which the novel compositions of the present invention may be included may be formulated for oral or injection administration and include aqueous solutions of suitably flavored syrups, aqueous or oily suspensions, and flavored emulsions with edible oils such as cottonseed oil, sesame oil, coconut oil or peanut oil, as well as elixirs and similar pharmaceutical vehicles.

Compositions for inhalation or inhalation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof and powders. Liquid and solid compositions may contain suitable pharmaceutically acceptable excipients as described above. Preferably, such compositions are administered by the oral or nasal respiratory route to elicit a local or systemic route.

effect. Compositions in preferably pharmaceutically acceptable solvents may be nebulized using inert gases. The atomized solutions may be inhaled directly from the nebulizer or the nebulizer may be attached to a face mask or intermittent positive pressure breathing apparatus. Solutions, suspensions, or powder compositions may be administered, preferably orally or nasally, from devices which administer the formulation in an appropriate manner.

The following formulation examples illustrate the pharmaceutical compositions of the present invention.

Example of preparation 1

Hard gelatin capsules containing the following ingredients were prepared.

Component

Quantity (mg / capsule)

Active ingredient Starch

Magnesium stearate

30.0 305.0 5.0

The above ingredients were mixed and filled into hard gelatin capsules in an amount of 340 mg.

Example of preparation 2

Tablets were prepared using the ingredients listed below:

Amount

...... '0-00 • 9 9 9 0 00 0 0 0 - 41 - ..... • 0. . '00 _: -00 •. . · · 0 0 «·· .. • 9 000000 · · 0 ^: '·····: 0 • 00 0 ·· 00 00 Ingredient (mg / tablet) Active ingredient 25.0 Cellulose, microcrystalline 200.0 Colloidal silica 10.0 Stearic acid 5.0 The ingredients were mixed and pressed into shape tablets of which

each weighing 240 mg.

Example of preparation 3

A dry powder inhalation preparation was prepared containing the following components:

Folder Weight %

Active ingredient

Lactose

The active ingredient is mixed with lactose and the mixture is placed in a dry powder inhaler.

Example of preparation 4

Tablets, each containing 30 mg of active ingredient, were prepared as follows:

Component

Quantity (mg / tablet) · · · · · ♦ ♦ ♦ mg mg mg mg mg mg mg mg mg mg mg mg mg mg mg ·· ··· ··· ·· · Φ

Active Ingredient 30.0 mg

Starch 45.0 mg

Microcrystalline cellulose35.0 mg

Polyvinylpyrrolidone (as a 10% solution in sterile water) 4.0 mg

Sodium carboxymethyl starch 4.5 mg

Magnesium stearate 0.5 mg

Talc_ 1.0 mg

Total 120 mg

The active ingredient, starch and cellulose are sieved through a 20 mesh U.S. sieve. mesh and mix thoroughly. The polyvinylpyrrolidone solution is mixed with the resulting powder, and sieved through a 16 mesh U.S. sieve. Mesh. The granules so formed are dried at 50-60 ° C and sieved through a 16 mesh U.S. sieve. Mesh. Sodium carboxymethyl starch, magnesium stearate and talc, pre-sieved through a 30 mesh U.S. sieve. mesh was then added to the granules, and after mixing, the substance was compressed on a tablet machine to give tablets each weighing 120 mg.

Example of preparation 5

Capsules, each containing 40 mg of drug, were formulated as follows:

Component

Quantity (mg / capsule)

· '

Active ingredient Starch

Magnesium stearate

40.0 mg 109.0 mg 1.0 mg

Overall

150.0 mg

The active ingredient, starch and magnesium stearate were mixed, sieved through a 20 mesh U.S. sieve. and filled into hard gelatin capsules in an amount of 150 mg.

Example of preparation 6

Suppositories, each containing 25 mg of active ingredient, were formed as follows:

Component Quantity

Active Ingredient 25 mg

Saturated fatty acid glycerides up to 2,000 mg

The active ingredient was passed through a 60 mesh U.S. sieve. and suspended in saturated fatty acid glycerides that have previously been melted using minimal heat. The mixture was then poured into a suppository mold with a nominal capacity of 2.0 g and allowed to cool.

Example of preparation 7

Suspensions containing 50 mg of the drug at a dose of 5.0 ml were formed as follows:

• ·

Component

Amount

Active Ingredient Xanthan gum

50.0 mg 4.0 mg

Sodium carboxymethylcellulose (11%) Microcrystalline cellulose (89%) 50.0 mg

Sucrose

Sodium benzoate Flavor and coloring agent Purified water up to

1,75 g

10.0 mg

q.v.

5.0 ml

The active ingredient, sucrose and xanthan gum were mixed, sieved through a 10 mesh U.S. sieve. and then mixed with a preformed solution of microcrystalline cellulose and sodium carboxymethylcellulose in water. Sodium benzoate, flavor, and color were dissolved in a small amount of water and added with stirring. Sufficient water was then added to reach the desired volume.

• «• ·· · r *» · '' · ¹ · '

··· ^: · · · ·

Example of preparation 8

Amount

Ingredient (mg / capsule)

Active ingredient 15.0 mg

Starch 407.0 mg

Magnesium stearate 3.0 mg

Total 425.0 mg

The active ingredient, starch and magnesium stearate were mixed, sieved through a 20 mesh U.S. sieve. mesh and filled into hard gelatin capsules at 560 mg.

Example of preparation 9

The subcutaneous preparation may be prepared as follows:

Component

Amount

Active ingredient 5.0 mg

Corn oil 1 ml

Example of preparation 10

A topical preparation may be prepared as follows:

• · »· · · ·

Component

Amount

Active Ingredient 1-10 g Emulsifying wax 30 g Liquid paraffin 20 g Soft white paraffin up to 100 g

The soft white paraffin was heated until it melted. Liquid paraffin and emulsifying wax were added and the mixture was stirred until dissolved. The active ingredient was added and stirring continued until dispersion was formed. The mixture was then cooled until it solidified.

Other preferred formulations utilizing the method of the present invention employ a transdermal delivery device ("patch"). Such transdermal patches may be used to provide continuous or discontinuous infusion of the compounds of the present invention in controlled amounts. The construction and use of transdermal patches for the administration of a pharmaceutical agent is well known in the art. See, for example, U.S. Pat. No. 5,023,252, issued June 11, 1991, which is incorporated herein by reference. Such patches may be designed to deliver the pharmaceutical agent in a continuous or burst manner or, if desired.

It is often desirable or necessary to deliver the pharmaceutical compositions to the brain, either directly or indirectly. Direct techniques typically involve placing a drug delivery catheter into a recipient ventricular system that bypasses the blood-brain barrier. One such implantable delivery system used to transport biological factors

• fc • • • · fc · • · · C • · fc · fc fc • · • • fc fc • • · fc · • · <· · • • • ·· · • · · • · E • • • fc fc · » • fc • · · • · · «· fc fc

to specific anatomical areas of the body is described in U.S. Pat.

No. 5,011,472, which is incorporated herein by reference.

Indirect techniques, which are generally preferred, typically involve preparing the composition to provide drug latency by converting the hydrophilic drug into a lipid soluble drug. Latency is usually achieved by blocking the hydroxyl, carbonyl, sulfate, and primary amine groups found in the compound to render the substance more lipid soluble and thus more suitable for transmission across the blood-brain barrier. Alternatively, the delivery of the hydrophilic drug may be enhanced by intra-arterial infusion of hypertonic solutions that temporarily open the blood-brain barrier.

Other suitable formulations for use in the present invention may be found in Remingron's Pharmaceutical Sciences, Mace Publishing Company, Philadelphia, PA, 17th ed. (1985).

Applicability

The compounds and pharmaceutical compositions of the present invention are useful for inhibiting the release of the βamyloid peptide and / or its synthesis and accordingly can be used for the treatment of Alzheimer's disease in mammals, including humans.

As mentioned above, the compounds described above are suitable for use in the variety of drug delivery systems mentioned above. In addition, in order to increase the half-life of the administered compound in serum, φ sloučeniny · · · · ·

Φ φ Φ Φ φ φ φ φ φ φ • • • • • • φ φ φ φ

The compounds may be encapsulated, inserted into the lumenal liposomes, prepared as a colloid, or treated by another conventional technique that achieves an extended half-life of the compound in serum. A number of methods that are useful for preparing liposomes are described, for example, in the patents of Szoka et al., U.S. Pat. Nos. 4,235,871, 4,501,728 and 4,837,028, all of which are incorporated herein by reference.

The amount of compound administered to a patient varies depending on the agent to be administered, the target of administration, such as prophylaxis or therapy, the condition of the patient, the mode of administration, and the like. For therapeutic administration, the compositions are administered to a patient already suffering from Alzheimer's disease in an amount sufficient to at least partially stop the further development of the disease symptoms and its complications. An amount suitable for achieving this goal is referred to herein as a "therapeutically effective dose." The amount effective for this use depends on the judgment of the attending physician and is based on factors such as the degree or severity of the Alzheimer's disease in the patient, the age, weight and general condition of the patient, and the like. Preferably, when used as a therapeutic agent, the compounds of the present invention are administered at dosages ranging from about 1 to about 500 mg / kg / day.

For prophylactic administration, the compositions are administered to a patient at risk of developing Alzheimer's disease (determined, for example, by genetic screening or family history) in an amount sufficient to inhibit the onset of symptoms of the disease. An amount suitable for achieving this goal is referred to herein as a &quot; prophylactically effective &quot;. in 9 '99 ·· • 9 · 9 9 9 9 9 99 99 9 · 9 · · 9999

9999 9 · 999,999

- 9 9 ^: 9 9 9 9 9

999 99 999 999 99 99 dose. The amount effective for this use depends on the judgment of the attending physician and is based on factors such as the degree or severity of Alzheimer's disease in the patient, the age, weight and general condition of the patient and the like. Preferably, when used as a prophylactic agent, the compounds of the present invention are administered in doses ranging from about 1 to about 500 mg / kg / day.

As mentioned above, the compounds are administered to a patient in the form of the pharmaceutical compositions described above. These compositions may be sterilized by conventional sterilization techniques or may be sterile filtered. The resulting aqueous solutions may be packaged for use as obtained, may be lyophilized, wherein the lyophilized formulation may be combined with a sterile aqueous carrier prior to administration. The pH of the compound preparation is typically between 3 and 11, more preferably from 5 to 9, and most preferably from 7 to 8. It will be appreciated that the use of some of said excipients, carriers or stabilizers will result in the formation of pharmaceutical salts.

The following synthetic and biological examples are provided to illustrate the present invention and are not intended to limit the scope of the present invention in any way. Unless otherwise stated, all temperatures are in degrees Celsius.

• · * · · · · · · · 0 · · · f.- 19? ·: · • 4

4

1 44

1. 1 4

4.9 4

499 499

4

44

DETAILED DESCRIPTION OF THE INVENTION

In the examples below, the following abbreviations have the meanings given below. If an abbreviation that is not defined is used, it has its generally accepted meaning.

aq = aqueous

Boc = tert-butoxycarbonyl

BOP = benzotriazol-1-yloxy-tris (dimethylamino) phosphonium hexafluorophosphate bd = broad doublet bs = broad singlet c = concentration (g / ml) cc = cubic centimeter

CDI = 1,1'-carbonyldiimidazole d = doublet dd = doublet doublet

DMAP = dimethylaminopyridine

DMF = dimethylformamide

DMSO = dimethylsulfoxide

EDC = 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride

EDTA = ethylene diamine tetraacetic acid eq. = equivalent

EtOAc = ethyl acetate

EtOH = ethanol g = gram h = hour

L = liter m = multiplet

M% = molar percentage

4. • 4 '7-9-' '4 ·. . 4 4 4 4: 4 4 4 ^; . 4, '' • 4 • 4 · · 4 · 4 4 '4 4 • • '- .. i'  9 · 4 . · 4 » 4 4 4 9 4 4 ' : 9 · ·· 999  4 4 4 4 4  i 4 ', ··· 4 • -4 --.- • 4 • 4 4 ' 44 4 4 4 4 4 4 4 4 4 4

max = maximum

MeOH = methanol meq - milli-equivalent mg - milligram ml = milliliter mm - millimeter mM = millimolar mmol = millimole mp = melting point

N = normal ng = nanogram nm = nanometer

OD = optical density

PEPC = 1- (3- (1-pyrrolidinyl) propyl) -3-ethylcarbodiimide pg = picogram pM = picomolar

General procedure A '

Coupling R ^X C (X *) (X) C (0) Cl with H ₂ NCH (R ² ) C (0) XR ³

To a stirred solution of alanine isobutyl ester hydrochloride (D, L) (from Example B below) (4.6 mmol) in 5 mL of pyridine was added 4.6 mmol of the acid chloride. Precipitation occurred immediately. The mixture was stirred for 3.5 hours, diluted with 100 mL diethyl ether, washed with 10% HCl three times, once with brine, 20% once with potassium carbonate, and once with brine. The solution was dried over magnesium sulfate, filtered and evaporated under reduced pressure to give the product. Other amino acid esters may also be used in this procedure.

Ftft ftft ftft ftftft ftftft ftft ftft ftftft ftftft ftftft ftft ftftft ftftft

General procedure B '

Coupling R ^X C (X ') (X) C (O) OH with H ₂ NCH (R ² ) C (O) XR ³

A solution of the acid (3.3 mmol) and CDI in 20 mL of THF was stirred for 2 hours. L-alanine, isobutyl ester, hydrochloride (from Example B below) (3.6 mmol) was added, followed by 1.5 mL (10.8 mmol) of acetylamine. The reaction mixture was stirred overnight. The reaction mixture was diluted with 100 mL diethyl ether, washed with 10% HCl three times, once with brine, 20% once with potassium carbonate, and once with brine. The solution was dried over magnesium sulfate, filtered and evaporated under reduced pressure to give the product. Other amino acid esters may also be used in this procedure.

General procedure C '

Esterification of R ¹ C (X ') (X) C (O) NHCH (R ² ) C (O) OH with HOR ³

To a stirred solution of phenylacetylvaline (1.6470 g, 7.0 mmol) in 20 mL of THF was added CDI (1.05 g, 6.5 mmol) and the mixture was stirred for 1.5 hours. 2-Methylbutanol (0.53 g, 6 mmol) was added to the mixture, followed by addition of NaH (0.16 g, 6.5 mmol). There was a bubbling immediately. The reaction mixture was stirred overnight. The reaction mixture was diluted with 100 mL diethyl ether, washed with 10% HCl three times, once with brine, 20% once with potassium carbonate, and once with brine. The solution was dried over magnesium sulfate, filtered and evaporated under reduced pressure to give the product. Other N-acyl amino acids and alcohols may also be used in this procedure.

General procedure D '

Hydrolysis of the ester to the free acid

4 44 4 4 4 4 4 4 4 4 4 • 4 4 4 4 4 4 4 4 • 44 4 4 4 4 4 4 • 4 4 4 4 4 4 444 4 4 4 4 4 4 4 4 4 4 • 4 4 4 4 4 4 4 4 4 4 4 4 4 4

Hydrolysis of the ester to the free acid was carried out by conventional methods. Below are two examples of such conventional de-esterification methods.

To the ester in a 1: 1 CH 3 OH / H 2 O mixture were added 2-5 equivalents of K ₂ CO ₃ . The mixture was heated to about 50 ° C for about 0.5 to 1.5 hours until tlc indicated completion of the reaction. The reaction mixture was cooled to ambient temperature and methanol was removed under reduced pressure. The pH of the remaining aqueous solution was adjusted to about 2 and ethyl acetate was added to extract the product. The organic phase was then washed with saturated aqueous NaCl and dried over MgSO 4. The solution was stripped of solvent under reduced pressure to give the product.

The amino acid ester was dissolved in dioxane / water (4: 1) to which LiOH (about 2 equiv.) Was added, which was dissolved in water so that the total solvent after addition was about 2: 1 dioxane: water. The reaction mixture was stirred until completion of the reaction and dioxane was removed under reduced pressure. The residue was diluted with EtOAc, the layers were separated and the aqueous layer acidified to pH 2. The aqueous layer was back extracted with EtOAc, the combined organics were dried over Na ₂ SO 4, and the solvent was removed under reduced pressure after filtration. The residue was purified by conventional means (for example, recrystallization).

The latter example will be further developed. 3NO ₂ Phenylacetyl alanine methyl ester 9.27 g (0.0348 mol) was dissolved in 60 ml dioxane and 15 ml H ₂ O and LiOH (3.06 g, 0.0731 mol) was added and dissolved in 15 ml H ₂ O. after stirring for 4 h the dioxane was removed under reduced pressure and

 '· 9 • · ·· « · ♦ • · 9 9 9 9 • · «« ' •  9 9 ♦ • · ·· •: 9 • ·· « «· · • · • • 9 • • • · · ·· • · · 99 9 • * • ·

the residue was diluted with EtOAc, the layers were separated and the aqueous layer was acidified to pH 2. The aqueous layer was back extracted with EtOAc (4 x 100 mL), the combined organics were dried over Na ₂ SO _4, and the solvent was removed under reduced pressure after filtration. The residue was recrystallized from EtOAc / isooctane to give 7.5 g (85%) of 3-nitrophenylacetylalanine.

C 11 H 12 N 2 O 5 gives C = 52.38, H = 4.80 and N = 11.11.

Found by analysis: C = 52.54, H = 4.85 and N = 11.08.

[?] 23 = - 29.9 at 589 nm.

General procedure E '

BOP coupling of acid and alcohol at low temperature A solution of methylene chloride containing carboxylic acid (100 mol%) and N-methylmorpholine (150 mol%) was cooled to -20 ° C under a nitrogen atmosphere. BOP (105 mol%) was added in one portion and the reaction mixture was maintained at -20 ° C for 15 minutes. The corresponding alcohol (120 mol%) was added and the reaction mixture was allowed to warm to ambient temperature and stirred for 12 hours. The reaction mixture was then poured into water and extracted with ethyl acetate (3x). The combined ethyl acetate was backwashed with saturated aqueous acid portions and the product was removed with citric sodium (2x) magnesium.

(2x), saturated brine (1x), aqueous bicarbonate dried over anhydrous or sodium sulfate, and the solvent was removed under reduced pressure to give crude crude. 1 1 · 1 · · · · · · · · · · · · · · · · · · · · · · · · · · · ·

General procedure F ^z

EDC coupling of acid and amine

The acid derivative was dissolved in methylene chloride. Amine (1 equiv.), N-methylmorpholine (5 equiv.) And hydroxybenzotriazole monohydrate (1.2 equiv.) Were added consecutively. The reaction mixture was cooled to about 0 ° C and then 1.2 equiv. of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride. The solution was left stirring overnight and returned to ambient temperature under N ₂ pressure. The reaction mixture was finally worked up by washing with a saturated aqueous solution of Na ₂ CO ₃ , 0.1 M citric acid and brine before drying the Na ₂ SO ₄ and removing the solvents to give the crude product. Pure products were obtained by flash chromatography in a suitable solvent.

General procedure G '

EDC coupling of acid and amine

The round bottom flask was charged with carboxylic acid (1.0 equiv), hydroxybenzotriazole hydrate (1.1 equiv) and amine (1.0 equiv) in THF under nitrogen atmosphere. An appropriate amount (1.1 equiv for free amines and 2.2 equiv. Of amine hydrochloride salt) of a base such as Hunig's base was added to a well stirred mixture, followed by EDC (1.1 equiv). After stirring for 4 to 17 hours at ambient temperature the solvent was removed under reduced pressure, the residue was taken up in EtOAc (or a similar solvent) / water. The organic layer was washed with saturated aqueous sodium bicarbonate, 1N HCl, brine, and dried over anhydrous sodium sulfate. In some cases, the isolated product was analytically pure at this stage, while in other cases it was necessary

9 9 9 '9' '99 - · ί ·

9999 99 99 999

9.9

9999 9 99999 999

9 9 9 9 9

999 99 999 999 99 99 purification by chromatography and / or recrystallization prior to biological experiments.

General procedure H '

Coupling R ^x C (X ') (X) C (O) C1 with H ₂ NCH (R ² ) C (0) XR ³

Excess oxalyl chloride in dichloromethane was added to the acid derivative along with one drop of DMF. The resulting mixture was stirred for about 2 hours or until bubbling ceased. The solvent was then removed under reduced pressure and diluted with anhydrous methylene chloride. To the resulting solution was added approximately 1.1 equiv. of a suitable amino acid ester and methylamine (1.1 equiv. in methylene chloride). The system was stirred at ambient temperature for 2 hours and then the solvent was removed under reduced pressure. The residue was dissolved in ethyl acetate, washed with 1 N HCl followed by 1 N NaOH. The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure to give the desired product.

General Procedure I '

P-EPC coupling

The P-EPC coupling uses an amino acid ester and a compound represented by substituted acetic acid. The acetic acid derivative is well known in the art and is typically commercially available. The amino acid ester is prepared by conventional methods from known and typically commercially available N-BOC amino acids as described in General Procedure J below.

• 00 · 0 '00 0 »00 0 0 00 00 · ·· ·

00 0 · · 0 · 0 00 00 0 0 0 000000 0 0 0 0 0 0 0 0

000 000 000 000 00 00

Specifically, the appropriate aminoester free base (0.0346 mmol) and substituted phenylacetic acid (0.069 mmol) were dissolved in 2.0 mL of CHCl 3 (EtOH-free), treated with 150 mg of P-EPC (0.87 meq / g) and reacted. the mixture was stirred for 4 days at 23 ° C. The reaction mixture was filtered through a pad of cotton, rinsed with 2.0 mL of GHCl 3 and the filtrate was evaporated under a stream of nitrogen. The purity of each sample was determined by ¹ H-NMR and ranged from 50% to> 95%. Between 8.0 and 15.0 mg of the final product was obtained from each reaction and tested without further purification.

General procedure J '

Synthesis of amino acid esters from the corresponding N-BOC amino acid

A. Esterification of acid.

The N-BOC amino acid was dissolved in dioxane and treated with excess alcohol (approximately 1.5 equiv.) And catalytic DMAP (100 mg) at 0 ° C. Stirring was continued until the reaction was complete and then the product was obtained by conventional means.

B. Removal of an N-BOC group.

The N-BOC protected amino acid was dissolved in methylene chloride (0.05M) and treated with 10 equivalents of TFA at ambient temperature under a nitrogen atmosphere. The reaction was monitored by thin layer chromatography until starting material was consumed usually in 1-5 hours. An additional 10 equivalents of TFA was added to the reaction mixture if the starting material was still present after 5 hours. The reaction mixture was carefully neutralized with Na ₂ CO 3, separated, the organic layer was washed with brine and dried over

• · ft · « • ft • ft • «J • · • · • · • • : · ' • ft • • • -ft • • • • • · • • • ftft · • ft • • • • • • • • • ft · • ft • · · • ftft • · «·

anhydrous. The crude amine was then used without purification.

Specific embodiments of these procedures are as follows:

1. Racemic (+/-) - N-BOC-α-amino-butyric acid (Aldrich) (9.29 g, 0.0457 mol) was dissolved in 100 ml dioxane and treated with isobutyl alcohol (6.26 ml, 0.0686 moles), EDC (5.72 g, 0.0457) and catalytic DMAP (100 mg) at 0 ° C. After stirring for 17 hours, the organics were evaporated under reduced pressure, the residue was diluted with EtOAc, washed with NaHCO 3, brine, and dried over Na 2 SO 4. Evaporation gave 8.42 g (71%) of an oil.

C 13 H 25 NO 4 gives: C = 60.21, H = 9.72 and N = 5.40.

Found: C = 59.91, H = 9.59 and N = 5.67.

The above amino acid N-BOC ester (8.00 g, 0.032 moles) was deprotected as described above to give 3.12 g (61%) of the free base as a colorless oil which solidified at rest.

2. LN-BOC-alanine (Aldrich) (8.97 g, 0.047 mol) was dissolved in 100 mL of CH 2 Cl 2, isobutyl alcohol (21.9 mL, 0.238 mol) and treated with DMAP (100 mg) and EDC (10, 0 g, 0.52 mol) at 0 ° C. The mixture was stirred for 17 hours, diluted with H 2 O, washed with 1.0 N HCl, NaHCO 3, then brine and dried over Na 2 SO ₄ . Filtration and evaporation gave 11.8 g (quantitatively) of LN-BOC alanine isobutyl ester, which was contaminated with a small amount of solvent. The sample was dried under vacuum for analytical analysis.

C12H23NO4 gives: C = 58.79, H = 9.38 and N = 5.71.

«0« 0

I 0

0 '0'

0 «« 1 • ·

0 · 0

0

Found: C = 58.73, H = 9.55 and N = 5.96.

The above amino acid N-BOC ester (11.8 g, 0.0481 mol) was deprotected as described above. The free base was converted to the corresponding HCl salt using saturated HCl (g) / EtOAc to give L-N-alanine, isobutyl ester, hydrochloride. 4.2 g (48%) of a colorless solid were obtained.

O7H15NO2. HCl gives: C = 46.25, H = 8.88 and N = 7.71.

Found: C = 46.01, H = 8.85 and N = 7.68.

General procedure K '

Formation of methyl ester from amino acid

The amino acid (amino acid or amino acid hydrochloride) was suspended in methanol and cooled to 0 ° C. HCl gas was bubbled through the solution for 5 minutes. The reaction mixture was allowed to warm to ambient temperature and then stirred for 4 hours. The solvents were then removed under reduced pressure to give the desired amino acid methyl ester. This product was usually further purified.

was obtained the hydrochloride used without

Example A '

Synthesis of free and polymer-bound PEPC

N-ethyl-N'-3- (1-pyrrolidinyl) propylurea

To a solution of 27.7 g (0.39 mol) of ethyl isocyanate in 250 ml of chloroform was added 50 g (0.39 mol) of 3- (1-pyrrolidinyl) propylamine dropwise with cooling. Once the addition was complete, the cooling bath was removed and the reaction mixture was stirred at ambient temperature for 4 hours. The reaction mixture was then concentrated under reduced pressure to afford 74.5 g (96.4%) of the desired urea as a clear oil.

1- (3- (1-pyrrolidinyl) propyl) -3-ethylcarbodiimide (P-EPC)

To a solution of 31.0 g (0.156 mol) of N-ethyl-N'-3- (1-pyrrolidinyl) propylurea in 500 ml of dichloromethane was added 62.6 g (0.62 mol) of triethylamine and the solution was cooled to 0 ° C. To this solution was then added 59.17 g (0.31 mol) of 4-toluenesulfonyl chloride in 400 mL of dichloromethane dropwise at such a rate that the reaction mixture was maintained at 0-5 ° C. After the addition was complete, the reaction mixture was warmed to ambient temperature and then heated to reflux for 4 hours. After cooling to ambient temperature, the reaction mixture was washed with saturated aqueous potassium carbonate (3 x 150 mL). The aqueous phases were combined and extracted with dichloromethane. All organic phases were combined and concentrated under reduced pressure. The resulting orange suspension was suspended in 250 mL of diethyl ether and the solution was separated from the solid by settling. The suspension / settling process was repeated three more times. The ethereal solutions were combined and concentrated under reduced pressure to give 18.9 g (67%) of the desired product as a crude orange oil. A portion of the oil was distilled in vacuo to give a colorless oil distilling at 78-82 ° C (0.4 mm Hg).

Preparation of polymer-borne form of 1- (3- (1-pyrrolidinyl) propyl) -3-ethylcarbodiimide (P-EPC)

Suspension of 8.75 g (48.3 mmol) of 1- (3- (1-pyrrolidin-yl) propyl) 3-ethylcarbodiimide and 24.17 g (24.17 mmol) of Merrifield resin (2% crosslinking, 200-400 mesh) , chloromethylated styrene / divinylbenzene copolymer, 1 equivalent (Cl / g) in dimethylformamide was heated at 100 ° C for 2 days. The reaction mixture was cooled and filtered, and the resulting resin was washed sequentially with 1 L of DMF, 1 L of THF and 1 L of diethyl ether. The remaining resin was then dried under vacuum for 18 hours.

Example B '

Preparation of alanine as isobutyl ester hydrochloride A mixture of 35.64 g (0.4 moles) of (D, L) -alanine (Aldrich) (or Lalanine (Aldrich)); 44 ml (0.6 moles) of thionyl chloride (Aldrich) and 200 ml of isobutanol were heated to reflux for 1.5 hours and the volatiles were completely removed on a rotary evaporator at 90 ° C under reduced pressure to give ( D, L) -alanine in the form of isobutyl ester hydrochloride (or L-alanine in the form of isobutyl ester hydrochloride) which was pure enough to be used for further transformations.

Example C '

Preparation of 3,5-dichlorophenylacetic acid

To a solution of 3.5 g of 3,5-dichlorobenzyl alcohol (Aldrich) in 75 ml of dichloromethane at 0 ° C was added 1.8 ml of methanesulfonyl chloride, followed by 3.5 ml of triethylamine added dropwise. After 2 hours, the solution was diluted to 150 mL with dichloromethane, washed with 3N HCl, saturated aqueous NaHCO 3, dried over Na ₂ SO _4, and the solvents removed to give the desired 3,5-dichlorobenzyl methanesulfonate as a yellow oil which was used without purification .

The crude sulfonate was dissolved in 50 mL DMF at 0 ° C and then 3 g KCN was added. After 2 hours, an additional 50 mL of DMF was added and the solution was stirred for 16 hours. The red solution was diluted with 1 L H ₂ O and acidified to pH 3 with 3N HCl. The aqueous solution was extracted with dichloromethane. The combined organics were washed with 3N HCl, dried over Na ₂ SO _4, and the solvents were removed under reduced pressure to give crude 3,5-dichlorophenylacetonitrile, which was used without purification.

Nitrile was added to a mixture of 40 ml of concentrated sulfuric acid and 50 ml of H ₂ O and heated to reflux for 48 hours, cooled to ambient temperature and stirred for 48 hours. The reaction mixture was diluted into 1 L of crushed ice, warmed to ambient temperature, and extracted with 2 x 200 mL dichloromethane and 2 x 200 mL ethyl acetate. Both sets of organics were combined and washed with saturated aqueous NaHCO 3. The NaHCO 3 fractions were combined and acidified to pH 1 with 3N HCl. The white solid was too fine to filter and was extracted with 2 x 200 mL dichloromethane. The combined organics were dried over Na ₂ SO ₄ and the solvents were removed under reduced pressure to give crude 3,5-dichlorophenylacetic acid as a white solid. The solid was suspended in hexane and filtered to give 1.75 g of a white solid.

NMR (CDCl ₃ ): (in ppm) 3.61 (s, 2H), 7.19 (s, 1H), 7.30 (s, 1H).

Example D '

Synthesis of N- (3-chlorophenylacetyl) alanine

The title compound was prepared using L-alanine (Nova Biochem) and 3-chlorophenylacetic acid (Aldrich) according to the general procedure F or G followed by hydrolysis using the general procedure D.

* ·> And ··

Example A1

Synthesis of N- (phenylacetyl) -D, L-alanine as isobutyl ester

Following the general procedure A above using phenylacetyl chloride (Aldrich) and D, L-alanine as isobutyl ester hydrochloride (from Example B above) the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by extraction with Et 2 O followed by washing with aqueous K 2 CO 3 and aqueous HCl.

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 7.23-7.36 (m, 5H), 6.18 (d, 1H), 4.58 (t, J = 7.3 Hz, 1H), 3 87 (m, 2H), 3.57 (s, 2H), 1.90 (m, 1H), 1.34 (d, J = 7.2Hz, 3H), 0.89 (d, J = 6.8 Hz, 6H). ¹³ C-nmr (CDCl 3): δ = 172.7, 170.3, 134.5, 129.2, 128.8,

127.2, 71.3, 48.1, 43.4, 27.5, 18.8, 18.3.

C15H21NO3 (MW = 263, 34; mass spectroscopy (MH ⁺ = 264))

Example A2

Synthesis of N- (3-phenylpropionyl) -D, L-alanine as isobutyl ester

Following the general procedure A above using 3-phenylpropionyl chloride (Aldrich) and D, L-alanine as isobutyl ester hydrochloride (from Example B above), the title compound was obtained as a solid, m.p. 51 ° -54 ° C. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by extraction with Et 2 O followed by washing with aqueous K 2 CO 3 and aqueous HCl.

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 7.25 (m, 2H), 7.19 (m, 3H), 6.28 (d, J = 7.2 Hz, 1H), 4.58 (quint) J = 7.2 Hz, 1H), 3.89 (m, 2H), 2.95 (t, J = 7.7 Hz, 2H), 2.50 (m, 2H), 1.92 ( m, 1H), 1.33 (d, J = 7.1 Hz, 3H), 0.91 (d, J = 6.7 Hz,

6H).

¹³ C-nmr (CDCl ₃ ): δ = 173.0, 171.5, 140.6, 128.3, 128.1,

126.0, 71.2, 47.8, 37.9, 31.4, 27.5, 18.79, 18.77, 18.3. C16H23NO3 (MW = 277.37, mass spectroscopy (MH ⁺ 278))

Example A3

Synthesis of N- (3-methylpentanoyl) -L-alanine as isobutyl ester

Following the procedure of General Procedure B using 3-methylpentanoic acid (Aldrich) and L-alanine as isobutyl ester hydrochloride (from Example B above), the title compound was prepared as an oil. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by extraction with Et 2 O followed by washing with aqueous K 2 CO 3 and aqueous HCl.

9 9

9 9 · 9

9

99

NMR data was as follows: 'H-nmr (CDC1 _3): δ = 6.08 (d, JJ = 7.3 Hz, IH), 3.92 (m, 2H), 3H), 1.40 (d, J = 7.2 Hz, 3H), 0.85-0.96 (m, 12H).

5.9 Hz, IH) 2.22 (m, IH) 1.35 (m, IH)

4.62 (quint., 1.84 to 2.00 (m, 1.20 (m, IH), ¹³ C-NMR (CDC1 _3): δ = 173.3, 172.1, 71.4, 47 , 9, 43.9, 32.3, 29.38, 29.35, 27, 6, 19.10, 19.06, 18.93, 15, 91, 18.72,

18.67, 11.3.

C13H25NO3 (MW = 243.35, mass spectroscopy (MH ⁺ 244))

Example A4

Synthesis of N - [(4-chlorophenyl) acetyl] -L-alanine as isobutyl ester

Following the procedure of General Procedure B using 4-chlorophenylacetic acid (Aldrich) and L-alanine as isobutyl ester hydrochloride (from Example B above), the title compound was prepared as a solid, m.p. 111-113 ° C. . The reaction was monitored by thin layer chromatography on silica gel and purification was performed by extraction with Et ₂ O followed by washing with aqueous K ₂ CC> 3 and aqueous HCl.

NMR data was as follows:

δ -nmr (CDCl ₃ ): δ = 7.30 (d, J = 8.2 Hz, 2H), 7.21 (d, J = 8.3 Hz, 2H), 6.18 (d, J = 5.5 Hz, 1H), 4.57 (quint., J =

7.2 Hz, 1H), 3.88 (m, 2H), 3.53 (s, 2H), 1.91 (m, 1H), 1.36 (d, J = 7.1 Hz, 3H) 0.90 (d, J = 6.8 Hz, 6H).

¹³ C-nmr (CDCl 3): δ = 172.8, 169.8, 133.1, 133.0, 130.6,

128.9, 71.4, 48.2, 42.6, 27.6, 18.85, 18.82, 18.4.

Ci ₅ H ₂ o ₃ O ₃ (MW = 297.78, mass spectroscopy (MH ⁺ 298)) · · · · · · · · · ·

Example A5

Synthesis of N - [(3,4-dichlorophenyl) acetyl] -L-alanine as isobutyl ester

Following the procedure of General Procedure B using 3,4-dichlorophenylacetic acid (Aldrich) and L-alanine as isobutyl ester hydrochloride (from Example B above), the title compound was obtained as a solid having a melting point of 81 ° C-83 ° C. Deň: 32 ° C. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by extraction with Et ₂ O followed by washing with aqueous K ₂ CO ₃ and aqueous HCl.

NMR data was as follows:

¹ H-nmr (CDC1 _3): δ = 0, 90 (d, J = 6.8 Hz, 6H), 1.38 (d, J = 7.1 Hz, 3H), 1.91 (m, 1H), 3.50 (s, 2H) 3.90 (m, 2H), 4.57 (quint. , J = 7.1 Hz, 1H), 6.31 (d, J = 4.9 Hz, 1H), 7.12 (m, lH) 7.38 (m, 2H) • ¹³ C-nmr (CDCl 3): δ = 18.4, 18.8 , 18,9, 27.6, 42.2, 48.3, 71,5, 128.6, 130.6, 131.2, 13; 1.3, 132 , 6, 134.7, 169.2,

172.8.

C ₁₅ H ₉ NO ₃ Cl ₂ (MW = 332.23, mass spectroscopy (MH ⁺ 332)

Example A6

Synthesis of N - [(4-methylphenyl) acetyl] -D, L-alanine as isobutyl ester

Following general procedure B using 4-methylphenylacetic acid (Aldrich) and D, L-alanine as isobutyl ester hydrochloride (from Example B above), the title compound was obtained as a solid, m.p. 102 ° C-104. Deň: 32 ° C. The reaction was monitored by thin layer chromatography on silica gel (Rf = 0.6 in 33% ethyl acetate / hexanes) and purification was 4 44

444 4 4 4 444 • · · · · 44 444 999 4 · .9 · - 4

4 4 4

4 was performed by extraction of Et ₂ O followed by washing with aqueous K ₂ CO ₃ and aqueous HCl.

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 0.90 (d, J = 6.7 Hz, 6H), 1.35 (d, J =

7.2 Hz, 3H), 1.91 (m, 1H), 2.34 (s, 3H), 3.55 (s, 2H), 3.88 (m, 2H), 4.58 (m, 1H), 6.05 (bd, 1H), 7.16 (s, 4H). ¹³ C-nmr (CDCl 3): δ = 18.5, 18.85, 18.87, 21.0, 27.6, 43.1,

48.1, 71.3, 129.2, 129, 6, 131.3, 136, 9, 170, 6, 172.8. C 16 H ₂₃ NO ₃ (MW = 277.37, mass spectroscopy (MH ⁺ 278))

Example A7

Synthesis of N - [(3-pyridyl) acetyl] -D, L-alanine as isobutyl ester

Following the procedure of General Procedure F using 3-pyridyl acetic acid hydrochloride (Aldrich) and D, L-alanine as isobutyl ester hydrochloride (from Example B above), the title compound was obtained as a solid, m.p. 62 °. C-64 ° C. The reaction was monitored by thin layer chromatography on silica gel (Rf = 0.48, 10% methanol / dichloromethane) and purification was by silica gel chromatography.

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 8.40 (d, J = 2.8, 2H); 7.6 (m, 1H):

7.16 (m, 2 H); 4.5 (quint., J = 7.2, 7.2, 1H); 3.8 (m, 2 H); 3.48 (s, 2 H); 1.8 (m, 1H); 1.30 (d, J = 7.2, 3H); 0.81 (d, J = 6.7, 6H).

¹³ C-nmr (CDCl ₃ ): δ = 173.4, 170, 1, 150, 6, 148.8, 137.4,

131.4, 124.1, 71.9, 48.9, 40.6, 28.1, 19.5, 19.4, 18.6. C ₄ H ₂ oN ₂ 0 ₃ (MW - 264, mass spectra MH ⁺ 265))

Example A8

Synthesis of N - [(1-naphthyl) acetyl] -L-alanine as isobutyl ester

Following the procedure of General Procedure B using 1-naphthylacetic acid (Aldrich) and L-alanine as isobutyl ester hydrochloride (from Example B above), the title compound was prepared as a solid having a melting point of 69 ° C-73 ° C. . The reaction was monitored by thin layer chromatography on silica gel and purification was performed by extraction with Et ₂ O followed by washing with aqueous K ₂ CO ₃ and aqueous HCl.

NMR data was as follows:

^* H-NMR (CDC1 ₃₎ δ = 0.83 (m, 6H) , 1.25 (d, J = 7.1 Hz, 3H), 1.81 (m, 1 H), 3.79 (m, 2H). 4.M (3s, 2H), 4.57 (quint., J = 7.3 Hz, 1H) 5.99 (d, J = 7.1) Hz, 1H), 7 , 44 l [m, 2H] 7.53 (m, 2H) 7.85 (m, 2H), 7.98 (m, 1H).

¹³ C-nmr (CDCl ₃ ): δ = 18.2, 18.81, 18.83, 27.5, 41.5, 48.2,

71.3, 123.7, 125.6, 126.1, 126.6, 128.2, 128.5, 128.7,

130.7, 132.0, 133.9, 170.3, 172.5.

C 19 H ₂₃ NO ₃ (MW = 313.40, mass spectroscopy (MH ⁺ 314))

Example A9

Synthesis of N - [(2-naphthyl) acetyl] -L-alanine as isobutyl ester

Following the procedure of General Procedure B using 2-naphthylacetic acid (Aldrich) and L-alanine as isobutyl ester hydrochloride (from Example B above), the title compound was prepared as a solid, m.p. 128 ° C-129 ° C. . The reaction was monitored by thin layer chromatography on silica gel and purification was performed

ΊΟ

• ·

4) extraction with Et ₂ O followed by washing with aqueous K ₂ CO ₃ and aqueous HCl.

NMR data was as follows:

^λ H-nmr (CDCl ₃ ): δ = 0.86 (m, 6H), 1.35 (d, J = 7.1 Hz, 3H), 1.78 (m, 1H), 3.76 (s 2H, 3.87 (m, 2H), 4.62 (quint., J =

7.2 Hz, 1H), 6.13 (d, J = 7.1 Hz, 1H), 7.41 (m, 1H), 7.48 (m, 2H), 7.74 (s, 1H) 7.83 (m, 3H).

¹³ C-nmr (CDCl ₃ ): δ = 18.4, 18.82, 18.85, 27.6, 43.7, 48.2,

71.4, 125.9, 126.3, 127.2, 127.6, 127.7, 128.2, 128.7,

132.0, 132.5, 133.5, 170.3, 172.8.

C19H23NO3 (MW = 313.40, mass spectroscopy (MH ⁺ 314)).

Example A10

Synthesis of N- (4-phenylbutanoyl) -L-alanine as isobutyl ester

Following the procedure of General Procedure B using 4-phenylbutanoic acid (Aldrich) and L-alanine as isobutyl ester hydrochloride (from Example B above), the title compound was prepared as an oil. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by extraction with Et ₂ O followed by washing with aqueous K 2 CO 3 and aqueous HCl.

NMR data was as follows:

¹ H-nmr (CDCl 3): δ = 0.92 (d , J = 6.7 Hz, 6H), 1.38 (d, J = 7.1 Hz, 3H), 1.96 (m, 3H); 2.21 (t, J = 7.1 Hz, 2H), 2.M (t, J = = 7.3 Hz, 2H), 3.90 (m, 2H), 4.59 (quint., J = 7.2 Hz, IH) 6.31 (d, 1H), 7.16 (m, 3H) 7.24 (m, 2H). ¹³ C-nmr (CDCl 3): δ = 18.3, 18.75, 18.78, 26, 8, 27 5, 34.9,

35.3, 47.8, 71.2, 125.7, 125.2, 128.3, 141.3, 172.1, 173.0. C ₁₇ H ₂₅ NO ₃ (MW = 291.39, mass spectroscopy (MH ⁺ 292)).

• O «44 '99. · · · · <6 · · ·

99 ····

9 • O 4 9

Example All

Synthesis of N- (5-phenylpentanoyl) -L-alanine as isobutyl ester

Following the procedure of General Procedure B using 5-phenylpentanoic acid (Aldrich) and L-alanine as isobutyl ester hydrochloride (from Example B above), the title compound was prepared as an oil. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by extraction with Et ₂ O followed by washing with aqueous K 2 CO 3 and aqueous HCl.

NMR data was as follows:

H-NMR (CDC1 _3): δ = 7.23 (m, 2H), 7.17 (m, 3H), 6.30 (d,

1H), 4.59 (quint., J = 7.3 Hz, 1H), 3.91 (m, 2H), 2.61 (t, J = 7.2 Hz, 2H), 2.22 (t J = 7.2 Hz, 2H), 1.93 (m, 1H), 1.66 (m, 4H), 1.38 (d, J = 7.2 Hz, 3H), 0.92 (d , J = 6.7

Hz, 6H).

¹³ C-nmr (CDCl ₃ ): δ = 173, 1, 172.3, 142.0, 128.2, 128.1,

125.6, 71.2, 47.8, 36.1, 35.5, 30.8, 27.5, 25.0, 18.80,

18.77, 18.4.

C18H27NO3 (MW = 305.39, mass spectroscopy (MH ⁺ 306)).

Example A12

Synthesis of N - [(4-pyridyl) acetyl] -D, L-alanine as isobutyl ester

Following the general procedure F and using 4-pyridyl acetic acid in the form of (Aldrich) and (D, L) -alanine hydrochloride in the form of isobutyl ester hydrochloride (from Example B above), the title compound was obtained as a solid having a melting point. Mp 64 ° C-66 ° C. The reaction was monitored by thin layer chromatography

• ·

Silica gel (Rf = 0.43 10% methanol / dichloromethane) and purification was by silica gel chromatography.

NMR data was as follows:

^x H-nmr (CDCl ₃ ): δ = 8.51 (dd, J = 1.6, 2.8, 1.6, 2H); 7.23 (dd, J = 4.3, 1.6, 4,4, 2H); 6.71 (d, J = 6.8, IH); 4,56 (quint., J = 7.3, 7,2, IH); 3.88 (m, 2H); 3.53 (with, 2H); 1.89 (m, 1H); 1.36 (d, J = 7.2, 3H); 0, - 88 (d, J = = 6.7, , 6B)

¹³ C-nmr (CDCl ₃ ): δ = 173.5, 169.3, 150.5, 144.4, 125, 1,

72, 1, 48, 9, 43.0, 28.2, 19.5, 19.5, 18.9.

C14H20N2O3 (MW = 264, mass spectroscopy (MH ⁺ 265))

Example A13

Synthesis of N- (phenylacetyl) -L-alanine as isobutyl ester Following general procedure B and using phenylacetyl chloride (Aldrich) and L-alanine as isobutyl ester hydrochloride (from Example B above), the title compound was prepared as 45 ° C-47 ° C. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by extraction with Et ₂ O followed by washing with aqueous K ₂ CO ₃ and aqueous HCl.

NMR data was as follows:

^x H-nmr (CDC1 _3): .Delta. = 7.24-7.39 (m, 5 H, 6.14 (d, 1H), 4.58 (t, J = = 7.3 Hz, 1H), 3.88 (m, 2H), 3.58 (s, 2H); 1.90 (m, IH), 1 35 (d, J = 7.2 Hz, 3H), 0.89 (d, J = 6.7Hz, 6H). ¹³ C-nmr (CDC1 _3): δ = 172.8, 170.4, 134.5, 129.3, 128.9, 127,2, 71.3, 48, 1, 43.5, 27.5, 18.9, 18.8, 18.4.

C ₁₅ H ₂₁ NO ₃ (MW = 263.34, mass spectroscopy (MH ⁺ 264)).

• 1 49

4 9 ·

4 4 ·

000 000 • 0

0 0 0

Example A14

Synthesis of 2 - [(3,4-dichlorophenyl) acetamido] butyric acid as isobutyl ester

Following the procedure of General Procedure I above and using

3,4-dichlorophenylacetic acid (Aldrich) and iso-butyl-2-aminobutyrate (prepared according to the general procedure J above) gave the title compound. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 7.36 (m, 3H), 6.03 (bd, 1H), 4.54 (m, 1H), 3.87 (m, 2H), 3.49 (s, 2H), 1.93 (m, 2H), 1.72 (m, 1H), 0.88 (d, 6H), 0.80 (t, 3H).

Example A15

Synthesis of 2 - [(3-methoxyphenyl) acetamido] butyric acid as isobutyl ester

Following the general procedure I above using 3-methoxyphenylacetic acid (Aldrich) and iso-butyl 2-aminobutyrate (prepared according to the general procedure J above), the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

¹ H-NMR (CDC1 _3): δ = 6.75 (m, 4H), 5.93 (bd, 1H), 4.51 (m, 1H), 3.83 (m, 2H), 3.75 (s, 2H), 3.52 (s, 2H), 1.82 (m, 2H), 1.60 (m, 1H), 0.64 (d, 6H), 0.74 (t, 3H) .

C17H25NO4 (MW = 307.39, mass spectroscopy (MH ⁺ 309)).

99 • • • 99 9 99

9 9

9 9 9

9 9 9

99,999

9

9 9 9

Example Α16

Synthesis of 2 - [(4-nitrophenyl) acetamido] butyric acid as isobutyl ester

Following general procedure I above and using 4-nitrophenylacetic acid (Aldrich) and iso-butyl 2-aminobutyrate (prepared according to general procedure J above), the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 8.16 (d, 2H), 7.44 (d, 2H), 6.04 (bd, 1H), 4.55 (m, 1H), 3.86 (m, 2H), 3.66 (s, 2H), 1.86 (m, 2H), 1.67 (m, 1H), 0.85 (d, 6H), 0.81 (t, 3H) .

C16H22N2O5 (MW = 322.36, mass spectroscopy (MH ⁺ 323)).

Example A17

Synthesis of 2 - [(3,4-methylenedioxyphenyl) acetamido] butyric acid as isobutyl ester

Following the procedure of General Procedure I above and using

3,4- (methylenedioxy) phenyl acetic acid (Aldrich) and isobutyl-2-aminobutyrate (prepared according to general procedure I above) gave the title compound. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 6.72 (m, 3H), 5.92 (bd, 1H), 4.54 (m, 1H), 3.86 (m, 2H), 3.66 (s, 2H), 1.86 (m, 2H), 1.66 (m, 1H), 0.89 (d, 6H), 0.79 (t, 3H).

Example A18

Synthesis of 2 - [(thien-3-yl) acetamido] butyric acid as isobutyl ester

Following general procedure I above and using 3-thiopheneacetic acid (Aldrich) and iso-butyl 2-aminobutyrate (prepared according to general procedure J above), the title compound was prepared. Reaction

was monitored by chromatography on thin layer on Silica gel and purification was performed filtration, how Yippee described in the general procedure. NMR data was as follows: H-NMR (CDC1 _3): δ = 7.37 (m, IH), 7.16 (m, 1H), 7.04 (m, 1H), 6.05 (Μ, 1H), 4.57 (m, 1H), 3.66 (s, 2H), 1.93 (m, 2H), 1.67 (m, 1H), 0.91 (d, 6H), 0.86 (t, 3H). Example A19 Synthesis of 2 - [(4-chlorophenyl) acetamido] butyric acid ve

isobutyl ester

Following the general procedure I above using 4-chlorophenylacetic acid (Aldrich) and iso-butyl 2-aminobutyrate (prepared according to the general procedure J above), the title compound was prepared. The reaction was monitored by thin layer chromatography on

silica gel and cleaning was carried out filtration, how Yippee described in the general procedure. NMR data was following: ^* H-NMR (CDC1 ₃₎ δ = 7.22 (m, 2H), 7.11 (m, 2H) 5, 80 (m, 1H), 4.44 (m, 1H), 3.78 (m, 2H), 3.43 (s, 2H) 1.77 (m, 2H), 1.56 (m, 1H), 0.83 (d, 6H), 0.71 (t, 3H).

Φ ΦΦ * · e · · · φ ·

Φ · ♦ · φ · · · · · · · · · · · · ·

Example Α20

Synthesis of 2 - [(3-nitrophenyl) acetamido] butyric acid as isobutyl ester

Following the general procedure I above using 3-nitrophenylacetic acid (Aldrich) and iso-butyl-2-aminobutyrate (prepared according to the general procedure I above), the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

1 H-NMR (CDCl ₃ ): δ = 8.15 (m, 2H), 7.65 (m, 1H), 6.08 (m,

1H), 4.46 (m, 1H), 3.92 (m, 2H), 3.68 (s, 2H), 1.91 (m,

2H), 1.75 (m, 1H), 0.98 (d, 6H), 0.71 (t, 3H).

Example A21

Synthesis of 2 - [(2-hydroxyphenyl) acetamido] butyric acid as isobutyl ester

Following the procedure of General Procedure I above and using

2-hydroxyphenylacetic acid (Aldrich) and iso-butyl-2-aminobutyrate (prepared according to the general procedure J above) prepared the title compound. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

^x H-nmr (CDCl ₃ ): δ = 7.14 (m, 1H), 7.01 (m, 1H), 6, 93 (m, 1H), 6.79 (m, 1H), 6.46 (m, 1H), 4.51 (m, 1H), 3.87 (m, 2H), 3.57 (s, 2H), 2.01 (m, 2H), 1.75 (m, 1H), 0.89 (d,

6H), 0.85 (t, 3H).

• · ·· • ·· • 4 ·· • • · • · • • · ♦ • • 4 • 9 • ·

♦ · · * * * ♦ ♦ ♦ ♦ ♦ ♦ ♦

Example A22

Synthesis of 2 - [(2-naphthyl) acetamido] butyric acid as isobutyl ester

Following the procedure of General Procedure I above and using

Of 2-naphthylacetic acid (Aldrich) and iso-butyl-2-aminobutyrate (prepared according to the general procedure J above) gave the title compound. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 7.83 (m, 7B), 5.95 (m, 1H), 4.58 (m, 1H), 3.84 (m, 2H), 3.75 (s, 2H), 1.89 (m, 2H), 1.63 (m, 1H), 0.91 (d, 6H), 0.81 (t, 3H).

C20H25NO3 (MW = 327.42, mass spectroscopy (MH ⁺ 328)).

Example A23

Synthesis of 2 - [(2,4-dichlorophenyl) acetamido] butyric acid as isobutyl ester

Following the procedure of General Procedure I above and using

2,4-dichlorophenylacetic acid (Aldrich) and iso-butyl-2-aminobutyrate (prepared according to General Procedure J above) prepared the title compound. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 7.49 (m, IH), 7.22 (m, 2H), 5.98 (m,

1H), 4.52 (m, 1H), 3.86 (m, 2H), 3.61 (s, 2H), 1.84 (m,

2H), 1.62 (m, 1H), 0.87 (d, 6H), 0.80 (t, 3H).

Example A24

Synthesis of 2 - [(4-bromophenyl) acetamido] butyric acid as isobutyl ester

Following general procedure I above and using 4-bromophenylacetic acid (Aldrich) and iso-butyl 2-aminobutyrate (prepared according to general procedure J above), the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

^λ H-nmr (CDCl ₃ ): δ = 7.43 (d, 2H), 7.19 (d, 2H) 5.85 (m, 1H), 4.51 (m, 1H), 3.81 (m, 2H), 3.47 (s, 2H), 1.84 (m, 2H), 1.61 (m, 1H), 0.84 (d, 6H), 0.76 (t, 3H). C ₁₆ H ₂₂ NO ₃ Br (MW = 356.26, mass spectroscopy (MH ⁺ 358)).

Example A25

Synthesis of 2 - [(3-chlorophenyl) acetamido]) butyric acid as isobutyl ester

Following the procedure of General Procedure I above and using

Of 3-chlorophenylacetic acid (Aldrich) and iso-butyl-2-aminobutyrate (prepared according to general procedure I above), the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was following: ¹ H-nmr (CDCl ₃ ) δ = 7.25 (m) 3H), 7.12 (m, IH) 5.80 (m, 1H), 4.52 (m, 1H), 3.86 (m, 2H), 3.50 (with, 2H), 1.87 (m, 2H), 1.67 (m, 1H) 0.88 (d, 6H) 0.77 (t, 3H). C ₁₆ H ₂₂ NO ₃ C1 (MW = 311.81, mass spectroscopy (MH ⁺ 313)).

Example A26

Synthesis of 2 - [(3-fluorophenyl) acetamido] butyric acid as isobutyl ester

Following the general procedure 1 above and using

Of 3-fluorophenylacetic acid (Aldrich) and iso-butyl-2-aminobutyrate (prepared according to the general procedure J above) prepared the title compound. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 7.31 (m, IH), 7.01 (m, 3H) 5.95 (m, 1H), 4.54 (m, 1H), 3.84 (m, 2H), 3.54 (with, 2H), 1.88 (m, 2H), 1.65 (m, 1H), 0.87 (d, 6H), 0.81 (t, 3H). C ₁₆ H ₂₂ NO ₃ F (MW = 295.35, mass spectroscopy) (MH ⁺ 296)).

Example A27

Synthesis of 2 - [(benzothiazol-4-yl) acetamido] butyric acid as isobutyl ester

Following the procedure of General Procedure I above and using

4-Benzothiazol-4-yl acetic acid (Chemservice) and isobutyl-2-aminobutyrate (prepared according to the general procedure J above) prepared the title compound. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

δ -nmr (CDCl ₃ ): δ = 7.82 (m, 1H), 7.51-7.21 (m, 4H), 5.84 (m, IH), 4.51 (m, IH), 3 90 (s, 2H), 3.79 (m, 2H), 1.78 (m, 2H), 1.58 (m, 1H), 0.80 (d, 6H), 0.66 (t, 3H) ).

• ·· • ft ··· »··>

Ft ft ft ft ft ft ft ft ft ft ft ft ft ft ft ft

Example A28

Synthesis of 2 - [(2-methylphenyl) acetamido] butyric acid as isobutyl ester

Following the procedure of General Procedure I above and using

2-methylphenylacetic acid (Aldrich) and iso-butyl-2-aminobutyrate (prepared according to the general procedure J above) prepared the title compound. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

δ -nmr (CDCl ₃ ): δ = 7.18 (m, 410, 5.79 (m, lip, 4.54) (m, (m, 1H), 3.85 (m, 2H), 1.59 (m, 2H), 3.59 (s, 2H), 3.29 (s, 2H); 1H) 0.87 (d, 6H), 0.77 (t, 3H) 291.39, mass spectroscopy 3H), 1.51 (M ⁺ 291)). C17H25NO3 (MW = Example A29 Synthesis 2 - [(2-fluorophenyl) acetamido] butyric acid ve

isobutyl ester

Following the procedure of General Procedure I above and using

Of 2-fluorophenylacetic acid (Aldrich) and iso-butyl-2-aminobutyrate (prepared according to the general procedure J above) prepared the title compound. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

* · ♦ •

9 9 · • * * «» 9 9 9

999 999

999 999

99

¹ H-nmr (CDCl ₃ ) : δ = 7.25 (m, 1H), 7.09 (m, 3H) 6.03 (m, 1H), 4.54 (m, 1H), 3.87 (m, 2H), 3.57 (with, 2H), 1.89 (m, 2H), 1.64 (m, IH) 0.88 (d, 6H) 0.80 (t, 3H).

Example Α30

Synthesis of 2 - [(4-fluorophenyl) acetamido] butyric acid as isobutyl ester

Following the procedure of General Procedure I above and using

Of 4-fluorophenylacetic acid (Aldrich) and iso-butyl-2-aminobutyrate (prepared according to the general procedure J above) prepared the title compound. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

¹ H-NMR (CDC1 _3): δ = 7.20 (m, 2H), 6.97 (m, 2H) 5.87 (m, 1H), 4.492 (m, 1H), 3.83 (m, 2H), 3.48 (with, 2H), 1.86 (m, 2H), 1.60 (m, 1H) 0.87 (d, 1H); 6H), 0.78 (t, 3H). C ₁₆ H ₂₂ NO ₃ F (MW = 295.35 mass) spectroscopy (MH ⁺ 296)).

Example A31

Synthesis of 2 - [(3-bromophenyl) acetamido] butyric acid as isobutyl ester

Following the procedure of General Procedure I above and using

Of 3-bromophenylacetic acid (Aldrich) and iso-butyl-2-aminobutyrate (prepared according to the general procedure J above) prepared the title compound. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 7.45 (m, 2H), 7.23 (m, 2H), 5.95 (m, IH), 4.55 (m, IH), 3.84 (m 2H) 3.55 (s, 2H), 1.89 (m, 2H), 1.68 (m, 1H) 0.91 (d, 6H), 0.81 (t, 3H).

C ₁₆ H ₂₂ NO ₃ Br (MW = 356.26, mass spectroscopy (M ⁺ 357)).

Example A32

Synthesis of 2 - [(3-trifluoromethylphenyl) acetamido] butyric acid as isobutyl ester

Following the procedure of General Procedure I above and using

3-Trifluoromethylphenylacetic acid (Aldrich) and iso-butyl 2-aminobutyrate (prepared according to the general procedure J above) prepared the title compound. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 7.52 (m, 1H), 7.47 (m, 2H) 6.01 (m, 1H), 4.56 (m, 1H), 3.86 (m, 2H), 3.61 (with, 2H), 1.84 (m, 2H), 1.62 (m, 1H) 0.87 (d, 6H) 0.80 (t, 3H). C17H22NO3F3 (MW = 345, 36, mass spectroscopy) (MH ⁺ 345)) *

Example A33

Synthesis of 2 - [(2-thienyl) acetamido] butyric acid as isobutyl ester

Following general procedure I above and using 2-thiopheneacetic acid (Aldrich) and iso-butyl 2-aminobutyrate (prepared according to general procedure J above), the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 6.89 (m, 3H), 6.07 (bd, 1H), 4.50 (m, 1H), 3.82 (m, 2H), 3.71 (s, 2H), 1.85 (m, 2H), 1.62 (m, 1H), 0.81 (d, 6H), 0.75 (t, 3H).

C 14 H 21 NO 3 S (MW = 283.39, mass spectroscopy (MH ⁺ 284)).

Example A34

Synthesis of 2- (phenylacetamido) butyric acid as isobutyl ester

Following General Procedure H above and using phenylacetic acid (Aldrich) and iso-butyl 2-aminobutyrate (prepared according to General Procedure J above), the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel and the purification was by silica gel chromatography using 9: 1 toluene: EtOAc as the eluent.

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 7.17-7.28 (m, 5H), 6.23 (bd, 1H), 4.51 (m, 1H), 3.86 (m, 2H) 3.54 (s, 2H), 1.87 (m, 2H), 1.62 (m, 1H), 0.87 (d, 6H), 0.78 (t, 3H).

C16H23NO3 (MW = 277.36, mass spectroscopy (MH ⁺ 277)).

Example A35

Synthesis of N- (phenylacetyl) valine as 2-methylbutyl ester

Step A. Preparation of N- (phenylacetyl) valine

To a stirred solution of 5.15 g (44 mmol) of valine (Bachem) in 50 mL (100 mmol) of 2N NaOH cooled to 0 ° C was added dropwise 5.3 mL (40 mmol) of phenylacetyl chloride (Aldrich). The colorless oil precipitated. The reaction mixture was allowed to warm to ambient temperature and was stirred for

9 9

9 9 9 ft · 9 9 9

9 9 9

999 999

999 99 hours, washed with 50 mL diethyl ether, acidified to pH 23 with aqueous HCl. The white precipitate formed was filtered off, washed thoroughly with water followed by diethyl ether to give 7.1 g (30 mmol, 69% yield) of the title compound.

NMR data was as follows:

^x H-nmr (DMSO-d _6): δ = 12.63 (s, 1H), 8.25 (d, J = 8.6 Hz, 1H), 7 27 (m, 5H), 4.15 (m, 1H); 3.56 (d, J = 13.8 Hz, 1 H), 3.47 (d , J = 13.8 Hz, 1H), 2.05 (m, 1H), 0.87 (d, J = 6.8, Hz, 3H) 0.84 (d, J = 6.8 Hz, 3) ¹³ C-nmr (DMSO-d 6): δ = 173.2, 170.4, 136, 6, 129.0, 128.2,

126.3, 57.1, 41.9, 30.0, 19.2, 18.0.

C13H17NO3 (MW = 235.29, mass spectroscopy (MH ⁺ = 236))

Step B. Synthesis of N- (phenylacetyl) valine as 2-methylbutyl ester

Following the procedure of General Procedure C using the N (phenylacetyl) valine prepared in Step A above and 2-methylbutan-1-ol (Aldrich), the title compound was prepared as a diastereomeric mixture. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

@ ¹ H-nmr (CDCl3): .delta 7,25-7,40 (m, 5H) 5, 95 (d, 1H), 4,56 (m, 1H), 3.84-4.00 (m, 2H) 3.61 (s, 2H), 2.10 (m, 1H), 1.68 (m, 1 H), 1.38 (m, 1 H), 1, 15 (m, 1H) , 0.82-0.94 (m, 9H),

0.76 (d, 3H).

¹³ C-nmr (CDCl 3): δ = 171.84, 171.81, 170.7, 134.6, 129.31,

129.27, 128.9, 127.3, 69.8, 57.0, 43, 7, 33, 9, 31, 3, 25, 9,

25.8, 18.9, 17.4, 16.34, 16.27, 11.12, 11.07.

00 «0 00 ·· ••• 0 00 00 0000

00 0 0 0 0 .0 0 0 0 0 0 0 0 0,000,000 • 00 0 0 0 0

C18H27NO3 (MW = 305.42, mass spectroscopy (MH ⁺ 306)).

Example A36

Synthesis of N- (phenylacetyl) -L-methionine as isobutyl ester

L-Methionine (0.1299, 0.869 mmol) (Aldrich) was taken up in dioxane (5.0 mL) and treated with saturated sodium bicarbonate solution (5.0 mL) followed by phenylacetyl chloride (Aldrich) (0.114 mL, 0.822 mmol). After stirring for 17 hours at ambient temperature, the mixture was diluted with ethyl acetate, the layers were separated, and the aqueous layer was acidified to pH 2 with 5N HCl. The crude product was extracted into ethyl acetate, dried over sodium sulfate, dried in vacuo and used without further purification. N-phenylacetyl-L-methionine (0.1285 g, 0.447 mmol) was dissolved in 3.0 mL dioxane and isobutyl alcohol (0.2 mL) and treated with EDC (0.094 g, 0.492 mmol) and catalytic DMAP (0.015 g) ). After stirring for 17 hours at 23 ° C, the mixture was concentrated in vacuo to an oil, the residue was diluted in EtOAc and washed with 0.1 N HCl and saturated sodium bicarbonate. Silica gel chromatography using 98: 2 CHCl ₃ / MeOH as the eluent gave the pure product.

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 7.4-7.23 (m, 5H), 6.14 (bd, 1H), 4.70 (m, 1H), 3.89 (d, 2H) 3.62 (s, 2H), 2.43 (m, 2H), 2.12 (m, 1H), 1.93 (m, 2H), 0.94 (d, 6H).

Ci ₇ H ₂₅ NO ₃ S (MW = 323, 17, mass spectroscopy (M ⁺ 323)) · dioxane (5.0 ml) bicarbonate

Example A37

Synthesis of N- (phenylacetyl) -L-leucine as isobutyl ester

L-Leucine (Aldrich) (0.1149, 0.869 mmol) was taken up in and treated with saturated sodium solution (5.0 mL) followed by phenylacetyl chloride (Aldrich) (0.114 mL, 0.822 mmol). After stirring for 17 hours at ambient temperature, the mixture was diluted with ethyl acetate, the layers were separated and the aqueous layer was acidified to pH 2 with 5N HCl. The crude product was extracted into ethyl acetate, dried over sodium sulfate, dried in vacuo and used without further purification. N-Phenylacetyl-Lleucine (0.0081 g, 0.038 mmol) was dissolved in 2.0 mL of CHCl 3 (EtOH-free) and isobutyl alcohol (0.055 mL) and treated with P-EPC (100 mg, 0.87 milliequivalents). The mixture was stirred for 4 days, filtered through a pad of cotton, and the filtrate was evaporated under reduced pressure to an oil that was sufficiently pure for testing.

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 7.22 (m, 5H), 5.57 (d, 1H), 4.35 (m,

1H), 3.35 (m, 3H), 1.35 (m, 4H), 0.68 (m, 9H).

C18H27NO3 (MW = 305, 40, mass spectroscopy (M ⁺ 305)).

Example A38

Synthesis of N - [(3-chlorophenyl) acetyl] alanine as 3-methylbut-2-enyl ester

Following the general procedure C above and using N- (3-chlorophenylacetyl) alanine (from Example D above) and 3-methylbut-2-en-1-ol (Aldrich), the title compound can be prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by liquid chromatography using 30% EtOAc / hexane as the eluent.

9 · · ♦ · ♦ «• • •

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 7.39-7.16 (m, 4H), 6.06 (d,

5.29 (m, 1H), 4.63 (d, J = 9Hz, 2H), 3.56 (s,

1H), 5.382H), 1.79 (s, 3H), 1.7 (s, 3H), 1.39 (d, J = 9Hz, 3H)

Example A39

Synthesis of N - [(3-chlorophenyl) acetyl] alanine as cyclopropylmethyl ester

Following the general procedure C above and using N- (3-chlorophenylacetyl) alanine (from Example D above and cyclopropylmethanol (Aldrich), the title compound can be prepared. The reaction was monitored by thin layer chromatography on silica gel and purification by liquid chromatography using 3: 7 EtOAc: hexane as the eluent.

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 7.2-7.1 (m, 4H), 6.09 (s, 1H), 4.6 (dq, J = 9 Hz, 1H), 3.96 (dd, J = 9 Hz, 2H), 3.59 (s, 2H), 1.2 (d, J = 9 Hz, 3H), 1.2-1.0 (m, 1H), 0.60-0 50 (m, 2H); 0.300.20 (m, 2H).

Example A40

Synthesis of N - [(3-chlorophenyl) acetyl] alanine as 2-thienylmethyl ester

Following the general procedure C above using N- (3-chlorophenylacetyl) alanine (from Example D above and 2-thiophenemethanol (Aldrich), the title compound can be prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed liquid chromatography using 3: 7 EtOAc: hexane as the eluent.

• 4

4 4 4 ► 4 4 4

444 444

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 7.37-6, 97 (m, 7H), 5.97 (q, J = 14 Hz,

2H), 4.6 (dq, J = 9Hz, 1H), 3.76 (s, 2H), 1.38 (d, J =

9Hz, 3H).

Example A41

Synthesis of N - [(3-chlorophenyl) acetyl] alanine as (1-methylcyclopropyl) methyl ester

Following the general procedure C above using N- (3-chlorophenylacetyl) alanine (from Example D above and (1-methylcyclopropyl) methanol (Aldrich) the title compound can be prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by liquid chromatography using 3: 7 EtOAc: hexane as the eluent.

NMR data was as follows:

^λ H-nmr (CDCl ₃ ): δ = 8.6 (bd, J = 9 Hz, 1H), 3.86 (q, J = 14 Hz, 2H), 3.4 (s, 2H), 2, 29 (q, J = 9Hz, 1H), 1.3 (d, J = 9Hz, 3H), 1.03 (s, 3H), 0.5-0.4 (m, 2H), 0.4 -0.28 (m, 2 H).

Example A42

Synthesis of N - [(3-chlorophenyl) acetyl] alanine as 3-thienylmethyl ester

Following the general procedure C above and using N- (3-chlorophenylacetyl) alanine (from Example D above and 3-thiophenemethanol (Aldrich), the title compound can be prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed. liquid chromatography using 3: 7 EtOAc: hexane as the eluent.

NMR data was as follows:

• 9

9 9 9 9 • .9

9

9 9 9

9 9 • 9 9

999 999

¹ H-nmr (CDCl ₃ ): δ = 8.03 (bd, J = 9 Hz, IH) , 7.56-7.5 (m, 1H), 7.47 (bs, 1H), 7.4-7.17 (m, 4H), 7.06 (d, J = 9) Hz,

Δ), 5.1 (s, 2H), 4.3 (dq, 1H), 1.3 (d, J = 9 Hz, 3H).

Example A43

Synthesis of N - [(3-chlorophenyl) acetyl] alanine as 2-methylcyclopentyl ester

Following the general procedure C above and using N- (3-chlorophenylacetyl) alanine (from Example D above and 2-methylcyclopentanol (Aldrich), the title compound can be prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed liquid chromatography using 3: 7 EtOAc: hexane as the eluent.

NMR data was as follows:

H-NMR (CDC1 _3): δ = 7.39-7.16 (m, 4H), 6.3 (bd, IH), 4,79-

4.7 (m, IH), 4,6-4,25 (m, J = 9 Hz, 1H), 3.58 (s, 2H), 2,09 · -1.8 (m, 2H), 1.74 -1.6 (m, 2H), 1.39 (dd, J = 9 Hz, 3H), 1,2 (dt, J = 9 Hz IH), 0.98 (dd, J = 9Hz, 2H)

C17H22NO3Cl (MW = 323.52, mass spectroscopy (MH ⁺ 323)).

Example A44

Synthesis of N - [(3-chlorophenyl) acetyl] alanine as 2-methylprop-2-enyl ester

Following the general procedure C above and using N- (3-chlorophenylacetyl) alanine (from Example D above and 2-methylprop-2-en-1-ol (Aldrich), the title compound can be prepared. thin layer on silica gel and purification was performed by liquid chromatography using 3: 7 EtOAc: hexane as the eluent.

• · • ·

NMR data was as follows:

H-NMR (CDC1 _3): δ = 7.39-7.16 (m, 4H), 6.03 (bs, 1H), 4.77 (s, 2H), 4.7 to 4.29 (m 3H), 2.59 (s, 2H), 1.73 (s, 3H), 1.43 (d, J = 9Hz, 3H)

C15H18NO3Cl (MW = 295.76, mass spectroscopy (MH ⁺ 295)).

Example A45

Synthesis of N - [(3-chlorophenyl) acetyl] alanine as cyclohex2-enyl ester

Following the general procedure C above and using N- (3-chlorophenylacetyl) alanine (from Example D above and cyclohex-2-en-1-ol (Aldrich) the title compound can be prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by liquid chromatography using 3: 7 EtOAc: hexane as the eluent.

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 8.6 (bd, J = 9 Hz, 1H), 7.4-7.2 (m,

4H), 6.0-5.8 (m, 1H), 5.7-5.5 (m, 1H), 5.1 (bs, 1H), 4.134.29 (m, 1H), 3.5 (s, 2H), 2.1-1.9 (m, 2H), 1.8-1.69 (m,

1H), 1.69-1.49 (m, 4H), 1.3 (dd, J = 9Hz, 3H)

C17H20NO3Cl (MW = 321.8, mass spectroscopy (MH ⁺ 321.2)).

Example A46

Synthesis of N - [(2-phenylbenzoxazol-5-yl) acetyl] alanine as isobutyl ester

Following the procedure of General Procedure I above and using 5- (2-phenylbenzoxazol) -yl-acetic acid (CAS # 62143-69-5) and alanine as isobutyl ester (prepared according to the General Procedure I above), the compound was prepared from the title.

9-99 · 9 9 9 9 9

9 9 9 *

99 9 9 9999 • 9 99 9 9 · 999999 • 9 9 9

NMR data was as follows:

δ -nmr (CDCl ₃ ): δ = 8.24 (m, 3H), 7.68 (m, IH), 7.51 (m, 5H), 6.04 (m, 1H), 4.58 (m, 1H), 3.85 (m, 2H), 3, 68 (with, 2H), 1.9 (m, 1H), 1.35 (d, 1H); 3H) z 0.87 (d, 6H).

C22H24N2OZ) (MW = 380, mass spectroscopy (MH ⁺ 381)).

Example A47

Synthesis of N - [(3-methylthiophenyl) acetyl] alanine as isobutyl ester

Following the general procedure I above using 3-methylthiophenylacetic acid (CAS # 18698-73-2) and alanine as isobutyl ester (prepared according to the general procedure J above), the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 7.14 (m, 2H), 7.01 (m, 1H), 4.56 (m,

1H), 3.88 (m, 2H), 3.54 (s, 2H), 2.46 (s, 3H), 1.89 (m,

1H), 1.35 (d, 3H), 0.85 (d, 5H).

C16H23NO3S (MW = 309, mass spectroscopy (MH ⁺ 310)).

Example A48

Synthesis of N-4 - [(2-furyl) acetyl] alanine as isobutyl ester

Following the procedure of General Procedure I above and using 2-furylacetic acid (CAS # 2745-26-8) and alanine as isobutyl ester (prepared according to the General Procedure J above), the title compound was prepared. The reaction was monitored by thin layer chromatography on

• »· · 4

4 4 4 '' * * · · «· · · · ·

Silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 7.36 (m, IH), 6.34 (m, H), 6.21 (m, IH), 4.56 (m, IH), 3.91 (m, 2H), 3.61 (s, 2H), 1.92 (m, 1H), 1.38 (d, 3H) 0.89 (d, 6H).

C 13 H 19 NO 4 (MW = 253, mass spectroscopy (MH ⁺ 254)).

Example A4 9

Synthesis of N - [(benzofuran-2-yl) acetyl] alanine as isobutyl ester

Following General Procedure I above and using benzofuran-2-ylacetic acid (Maybridge) and alanine as isobutyl ester (prepared according to General Procedure J above), the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 7.51 (m, IH), 7.44 (m, 1H), 7.25 (m,

2H), 6.67 (s, 1H), 4.60 (m, 1H), 3.87 (m, 2H), 3.77 (s,

2H), 1.88 (m, 1H), 1.38 (d, 3H), 0.87 (d, 6H).

C17H21NO4 (MW = 303, mass spectroscopy (MH ⁺ 304)).

Example A50

Synthesis of N - [(benzothiophen-3-yl) acetyl] alanine as isobutyl ester

Following the procedure of General Procedure I above and using thianaphthen-3-ylacetic acid (Lancaster) and alanine in the form of isobutyl ester (prepared according to the General Procedure J above), the compound was prepared from ♦♦.

1 · φ · · · · · · · · · · · · · · · · · · · · · · 9 · · · · · · · · · · · · · · · · · · · · · 9 · · · · The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

^λ H-nmr (CDCl ₃ ): δ = 7.89 (m, IH), 7.76 (m, IH), 7.38 (m, 3H), 6.07 (m, 1H), 4.57 (m, IH), 3.92 (m, 2H), 3.82 (with, 4H), 1 M (m, 1H), 1.32 (d, 3H) 0.85 (d, 6H).

C17H21NO3S (MW = 319, mass spectroscopy (MH ⁺ 320)).

Example A51

Synthesis of N - [(2-chloro-5-thienyl) acetyl] alanine as isobutyl ester

Following the general procedure I above using 5-chloro-2-thienylacetic acid (CAS # 13669-19-1 and alanine as isobutyl ester (prepared according to the general procedure J above), the title compound was prepared. monitored by thin layer chromatography on silica gel and purification by filtration as described in the general procedure.

NMR data was as follows:

¹ H-NMR (CDC1 _3): δ = 6.77 (m, IH), 6.68 (d, IH), 6.31 (m,

1H), 4.59 (m, 1H), 3.91 (m, 2H), 3.38 (s, 2H), 1.90 (m,

1H), 1.39 (d, 3H), 0.89 (d, 5H).

C ₁₃ H ₈ NO ₃ SC 1 (MW = 303, mass spectroscopy (MH ⁺ 303)).

• · · · · · 9 9 9 9 9 9 9 9 9 9 9 9 9 44 9 4 9 9 4 9 4 4 4 9

49 9 9 4. 4 4 4

4,444 9 4-999 449

9 9 · · 9 9

944 44 444 449 49 94

Example A52

Synthesis of N - [(3-methylisoxazol-5-yl) acetyl] alanine as isobutyl ester

Following the procedure of General Procedure I above and using (3-methyl-isoxazol-5-yl) acetic acid (CAS # 19668-85-0) and alanine as isobutyl ester (prepared according to General Procedure J above), the following was prepared: the title compound. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

¹ H-nmr (CDC- ₁₃ ): δ = 6.07 (s, 2H), 4.56 (m, 1H), 3.92 (m,

2H), 3.68 (s, 2H), 2.29 (s, 3H), 1.94 (m, 1H), 1.89 (d, 3H) 0.91 (d, 6H).

C 13 H 20 N 2 O 4 (MW = 268, mass spectroscopy (MH ⁺ 269)).

Example A53

Synthesis of N - [(2-phenylthiothienyl) acetyl] alanine as isobutyl ester

Following the general procedure I above and using (2-phenyl-thiothienyl) acetic acid and alanine as isobutyl ester (prepared according to the general procedure J above), the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

1 H-NMR (CDCl ₃ ): δ = 7.21-7.11 (m, 6H), 6.92 (d, 1H), 4.56 (m, 1H), 3.87 (m, 2H), 3.72 (s, 2H), 1.94 (m, 1H), 1.38 (d, 3H) 0.89 (d, 6H).

C19C23NO3S2 (MW = 377, mass spectroscopy (MH ⁺ 378)).

44

4 4

4 4

444 444

44

Example A54

Synthesis of N - [(6-methoxybenzothiophen-2-yl) acetyl] alanine as isobutyl ester

Following the general procedure I above and using (6-methoxythianaphten-2-yl) acetic acid and alanine as the isobutyl ester (prepared according to the general procedure J above), the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

¹ H-NMR (CDC1 _3): δ = 7.59 (d, IH), 7.33 (d, IH), 7.16 (s, IH), 7.03 (dd, IH), 4.56 (m, 1H), 3.87 (s, 3H), 3.M (m, 2H), 3.76 (s, 2H), 1.85 (m, 1H), 1.30 (d, 3H) 0.86 (d, 6H). C18H23NO4S (MW = 3491, mass spectroscopy (MH ⁺ 350)).

Example A55

Synthesis of N - [(3-phenyl-1,2,4-thiadiazol-5-yl) acetyl] alanine as isobutyl ester

Following the general procedure I above and using (3-phenyl-1,2,4-thiadiazol-5-yl) acetic acid (CAS # 90771-06-5) and alanine as isobutyl ester (prepared according to the general procedure J above) the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

¹ H-NMR (CDC1 _3): δ = 7.47 (m, 5H), 4.66 (m, IH), 4.16 (s,

2H), 3.91 (m, 2H), 1.93 (m, 1H), 1.48 (d, 3H) 0.93 (d, 6H). C17H21N3O3S (MW = 347, mass spectroscopy (MH ⁺ 348)).

• 00 0 0 0 0 00 0 0 · 0 0 0 0 0 • 0 0 · • 00 0 0 0 « 0 0 • · 0 · 0 0 0 00 0 0 0 0 0 » 0 0 • 0 0 0 0 0 0 0 000 000 0 0 0 0

Example A56

Synthesis of N - [(2-phenyloxazol-4-yl) acetyl] alanine as isobutyl ester

Following General Procedure I above and using (2-phenyloxazol-4-yl) acetic acid (CAS # 22086-89-1) and alanine as isobutyl ester (prepared according to General Procedure J above), the compound was prepared from title. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

Example A57

Synthesis of N - [(3-methylphenyl) acetyl] alanine as isobutyl ester

Following the procedure of General Procedure I above and using 3-methylphenylacetic acid (Aldrich) and alanine as isobutyl ester (prepared according to the General Procedure J above), the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

δ -nmr (CDCl ₃ ): δ = 7.21 (m, 1H), 7.07 (m, 3H), 4.54 (m,

1H), 3.83 (m, 2H), 3.52 (s, 2H), 2.35 (s, 3H), 1.87 (m,

1H), 1.32 (d, 3H), 0.88 (d, 6H).

C16H23NO3 (MW = 277, mass spectroscopy (MH ⁺ 278)).

• »· ΦΦ

9 · · 9 9 9 9 · Φ 4 · - Φ φ φ::::::::

99 ΦΦ ΦΦΦ ΦΦΦ 99 99

Example Α58

Synthesis of Ν - [(2,5-difluorophenyl) acetyl] alanine as isobutyl ester

Following the procedure of General Procedure I above and using

2,5-difluorophenylacetic acid (Aldrich) and alanine as isobutyl ester (prepared according to the general procedure J above), the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

H-NMR (CDC1 _3): δ = 7.08-6.94 (m, 3H), 4.57 (m, 1H), 3.91 (m, 2H), 3.56 (s, 2H); 1.92 (m, 1H); 1.41 (d, 3H); 0.91 (d, 6H).

C ₁₅ H ₁₉ NO ₃ F ₂ 2 (MW = 299, mass spectroscopy (hlH + 300)).

Example A59

Synthesis of N - [(3,5-diflurophenyl) acetyl] alanine as isobutyl ester

Following the procedure of General Procedure I above and using

3,5-difluorophenylacetic acid (Aldrich) and alanine as isobutyl ester (prepared according to the general procedure J above), the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

δ -nmr (CDCl ₃ ): δ = 6.81 (m, 2H), 6.74 (m, 1H), 6.06 (m,

1H), 4.57 (m, 1H), 3.92 (m, 2H), 3.51 (s, 2H), 1.94 (m,

1H), 1.36 (d, 3H) 0.87 (d, 6H).

C ₁₅ H ₁₉ NO ₃ F ₂ (MW = 299, mass spectroscopy (MH ⁺ 300)).

• 0

0

0

0 0 ·· 00 ·

0 0 «'0' 000 ·» ·

Example A60

Synthesis of N - [(3-thienyl) acetyl] alanine as isobutyl ester

Following general procedure I above and using 3-thiopheneacetic acid (Aldrich) and alanine as isobutyl ester (prepared according to general procedure J above), the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

--nmr (CDCl ₃ ): δ = 7, 33 (m, 1H); . 14 (m, 1H) 7.01 (m, 1H), 6.5 (m, 1H), 4.58 (m, 1H), 3.88 (m, 2H), 3.60 (s, 2H) 1.91 (m, 1H); 1.37 (d, 3H), 0.92 (d, 6H) • Optical rotation: [a] 23 - 52 (c 1 MeOH) at 589 nm.

C 13 H 19 NO 3 S (MW-269, mass spectroscopy (MH ⁺ 269)).

Example A61

Synthesis of N - [(4-methylphenyl) acetyl] -L-alanine as isobutyl ester

Following the procedure of General Procedure I above using 4-methylphenylacetic acid (Aldrich) and L-alanine as isobutyl ester (prepared according to the General Procedure J above), the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by filtration as described in the general procedure.

NMR data was as follows:

• ·· «♦ · · · · <

· 99 99

90

0 9 9

0 9 9

999 999

9

9 9 99 ¹ H-nmr (CDCl ₃ ): δ = 7.11 (s, 4H), 5.93 (m, 1H), 4.58 (m,

1H), 3.58 (m, 2H), 3.54 (s, 2H), 2.33 (s, 3H), 1.89 (m,

1H), 1.32 (d, 3H), 0.89 (d, 6H).

C ₁₆ H 23 NO ₃ (MW = 277.35, mass spectroscopy (MH ⁺ 278)).

Example A62

Synthesis of N- (phenylacetyl) -L-alanine as S-1- (methoxycarbonyl) isobutyl ester

Following the general procedure K and using (S) - (+) - 2-hydroxy-2-methylbutyric acid (Aldrich) instead of the amino acid, methyl (S) - (+) - 2-hydroxy-2-methylbutyrate was prepared. Methyl (S) - (+) - 2-hydroxy-2-methylbutyrate was then coupled with carbobenzyloxy-L-alanine (Aldrich) using general procedure E to give carbobenzyloxy-L-alanine as S-1- (methoxycarbonyl) iso-butyl ester. Carbobenzyloxy-L-alanine as S1- (methoxycarbonyl) isobutyl ester (1.0 g) was then dissolved in 20 ml of methanol and 6N HCl (0.5 ml) and 10% palladium on activated carbon (0.1 g) was added. . The reaction mixture was hydrogenated at 40 psi hydrogen in a Parr apparatus for 5 hours at ambient temperature and then filtered through a pad of Celite. The filtrate was concentrated under reduced pressure to give L-alanine S-1- (methoxycarbonyl) as isobutyl ester hydrochloride (98% yield). L-Alanine S-1- (methoxycarbonyl) as isobutyl ester hydrochloride was then coupled with phenylacetic acid using general procedure G to give the title compound.

NMR data was as follows:

δ -nmr (CDCl ₃ ): δ = 7.35-7.20 (m, 5H), 6.22 (bd, 1H), 4.83 (d, 1H), 4.65 (p, 1H), 3.68 (s, 3H), 3.55 (s, 2H), 2.21 (m, 1H), 1.40 (d, 3H), 0.97 (d, 3H), 0.93 (d , 3H).

100 ALIGN!

• 0 • 0 00 00 00 * • 00 0 0 0 « 0 « • 00 • 0 • 0 0 '' 0 • 0 • · 0 «00« 0 0 0 - 0 0 • • « 0 0 • 00 • 0 • 0 0 000 0 0 0 0

¹³ C-NMR (CDC1 _3): δ = 173.25, 171, 18, 170.22, 135, 11, 129, 94, 129.50, 127.88, 52.67, 48.49, 43.98 , 30.53, 19.21, 18.75,

17.58.

Example A63

Synthesis of N - [(3-nitrophenyl) acetyl] -L-alanine as isobutyl ester

Following the procedure of General Procedure H above and using 3-nitrophenylacetic acid (Aldrich) and L-alanine as isobutyl ester hydrochloride (from Example B above), the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was carried out by recrystallization from butyl chloride.

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 8.17 (m, 2H), 7.68 (d, 1H), 7.52 (t,

1H), 6.18 (m, 1H), 4.48 (m, 1H), 3.94 (m, 2H), 3.67 (s,

2H), 1.93 (m, 1H), 1.42 (d, 3H), 0.91 (d, 3H).

Optical rotation: [a] ₂ 3 -49 (c 5, MeOH).

Example A64

Synthesis of N - [(3,5-difluorophenyl) acetyl] alanine as ethyl ester

Following general procedure G and using 3,5-difluorophenylacetic acid (Aldrich) and alanine as ethyl ester (Aldrich), the title compound was obtained as a solid, m.p. 93 ° C-95 ° C.

The reaction was monitored by thin layer chromatography on silica gel (Rf = 0.8 in EtOAC) and purification was performed by silica gel chromatography using EtOAc as the eluting solvent, followed by recrystallization from 1-chlorobutane. NMR data was as follows:

101 • ··. · ... ·· .. ····. . : .. 9 · 4> · ··. · '·:' .. ···· · • ·. · '99' 0 0 0 • · .0.0-. · .. 0. · • * 000 000 00 00 ^* H-NMR (DMSO-d _6): δ = 1.30 (d, 3H); 3.52 (s, 2 H).

C ₁₃ H N ₅ NO ₃ F ₂ (MW = 271.26, Mass Spectroscopy (MH ⁺ 271)).

Example A65

Synthesis of N - [(3-nitrophenyl) acetyl] methionine as ethyl ester

Following the general procedure G above and using 3-nitrophenylacetic acid (Aldrich) and methionine ethyl ester hydrochloride (Aldrich), the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was carried out by recrystallization from butyl chloride. NMR data was as follows: ¹ H-nmr (CDCl ₃ ): δ G = 8.18 (s, 1H), 8.15 (d, 1H) 7.66 (d,

1H), 7.48 (t, 1H), 6.30 (m, 1H), 4.67 (m, 1H), 4.21 (t, 2H), 3, 67 (with, 2H), 2.47 (t, 2H), 2.12 (m, 2), 2.08 (with, 3H), 1,27 (t, 3H). Optical rotation : [a] 23-30 (c5, MeOH).

Example A66

Synthesis of N - [(3-chlorophenyl) acetyl] alanine as isobutyl ester

Following General Procedure G above and using 3-chlorophenylacetic acid (Aldrich) and alanine as isobutyl ester (prepared according to General Procedure J above), the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel.

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 7.29 (m, 3H) 7. 18 (m, 1H), 6.0 (m, 1H), 4.56 (m, 1H), 3.89 (m, 2H), 3.53 (s, 2H), 1.91 (m, 1H), 1.39 (d, 3H), 0.91 (d, 3H).

Optical rotation: [α] -45 (c 5, MeOH).

C15H20NO3Cl (MW = 297.78, mass spectroscopy (MH ⁺ 297)).

Example A67

Synthesis of N - [(3-chlorophenyl) acetyl] alanine as 2- (N, N-dimethylamino) ethyl ester

Following the general procedure C above and using N- (3-chlorophenyl acetyl) alanine (from Example D above and 2- (N, N-dimethyl amino) ethanol (Aldrich), the title compound can be prepared. thin layer chromatography on silica gel and purification was performed by liquid chromatography using 0.1: 2: 0.79 NH 4 OH: EtOH: CHCl 3 as the eluent.

NMR data was as follows:

^X H- nmr (CDCl 3): 7.37 (s, 1H), 7.33-7.2 (m, 3H), 4.675-4.6 (m, 1H), 4.5-4.37 (m, 1H), 4.25-4.13 (m, 1H) 3.6 (d, J = 7) Hz, 2H), 2.86 (s, 2H) 2.3 (s, 6H), 1.23 (d, J = 9 Hz, 3 H).

C15H21N2O3Cl (MW = 313.799, mass spectroscopy (M ⁺ 313)).

Example A68

Synthesis of 2 - [(3,5-dichlorophenyl) acetamido] hexanoic acid as methyl ester

Following the procedure of General Procedure F above, using 3,5-dichlorophenylacetic acid (from Example C above) and L-norleucine as methyl ester hydrochloride (Bachem), the title compound was obtained as a solid having a melting point of 77 ° C. -78 ° C. The reaction was monitored by thin layer chromatography on silica gel (Rf = 0.70 in 40% EtOAC / hexanes) and purification was performed by silica gel flash chromatography using 40%

EtOAc / hexanes as the eluent.

'«·

0 0 0 0 · 0 · 0

103 - • 00 * 0 0 0 0 · 0 • 0 00 00 000 <00 • 00 00 0. 00 * « NMR data .a was following: ^x H-nmr (CDC1 ₃ ) : δ = 7.20 (s, , 7.18 (s), 6.6 (m), 4.55 (m), 3.7 (s) , 3,5 (s), 3.4 (s), 2.0 (s) , 1.8 (m), 1.6 (mj, 1,2 (m), 0 8 (t). ¹³ C-nmr (CDC1 ₃ ) : δ = 173.54, 169.67, 138.43, 135.72, 128 , 33,

128.07, 78.04, 77.62, 77.19, 53.Μ, 52.90, 43.14, 32.57,

27.87, 22.81, 14.41.

Example Α69

Synthesis of Ν - [(3,5-dichlorophenyl) acetyl] -L-alanine as isobutyl ester

Following the procedure of General Procedure F above and using

3,5-dichlorophenylacetic acid (from Example C above) and L-alanine as isobutyl ester hydrochloride (from Example B above) gave the title compound as a solid having a melting point of 1150-116 ° C. The reaction was monitored by thin layer chromatography on silica gel (Rf = 0.40 in 3% methanol / dichloromethane) and purification was performed by silica gel flash chromatography using 3% methanol / dichloromethane as the eluent.

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 7.27 (d, J = 2 Hz, IH), 7.19 (s, 2H)

6.22 (d, J = 6Hz, 1H), 4.59 (quint., J = 7Hz, IH), 3.9 (q, J = 4Hz, 2H), 3.5 (s, 2H) ), 1.9 (m, 1H), 1.4 (d, J = 7)

Hz, 3H), 0.91 (d, J = 7Hz, 6H).

¹³ C-NMR (CDC1 _3): δ = 173, 45, 169, 37, 138.31, 135, 75, 128.39, 128.11, 78.04, 77.61, 77.19, 72.19 , 54.03, 48.97, 43.12,

28.24, 19.52, 19.49, 19.09.

C15H19NO3Cl2 (MW = 331.9, mass spectroscopy (MH ⁺ 332)).

• φ φ

104

Example Α70

Synthesis of N- (cyclohexylacetyl) -L-alanine as isobutyl ester

Following the general procedure B above using cyclohexylacetic acid (Aldrich) and L-alanine as isobutyl ester hydrochloride (from Example B above), the title compound was obtained as a solid having a melting point of 92 ° C-93 Noc: 2 ° C. The reaction was monitored by thin layer chromatography on silica gel (Rf = 0.39 in 1: 3 EtOAc: hexane) and purification was performed by extraction with Et 2 O followed by washing with aqueous K 2 CO 3 and aqueous HCl.

NMR data was as follows:

¹ H-nmr (CDCl 3): δ = = 0.93 (d, J = 6.7 Hz, 6H), 0.85-1.01 (m, 2H), 1 05-1.35 (m 3H), 1.40 (d, J = 7.1) Hz, 3H), 1.60-1.85 (m, 7H) 1.95 (m, 1H), 2.06 (d, J = t 7.0 Hz, 2H), 3.92 (m, 2H), 4 61 (m, 1H); 6.08 (bd, IH). ¹³ C-nmr (CDCl 3): δ = 18.7, 18.9, 26.0, 26, 1, 27.6, 33.0, 35.3, 4 4.6, 47.9, 71.4, 171, 8, 173.3.

C15H27NO3 (MW = 269, 39, mass spectroscopy (MH ⁺ 270)).

Example A71

Synthesis of N- (cyclopentylacetyl) -L-alanine as isobutyl ester

Following the general procedure B above and using cyclopentylacetic acid (Aldrich) and L-alanine as isobutyl ester hydrochloride (from Example B above), the title compound was obtained as a solid having a melting point of 62 ° C - 64 ° C. Deň: 32 ° C. The reaction was monitored by thin layer chromatography on silica gel (R _f = 0.37 in 1: EtOAc: hexane) and purification was performed.

4 4 9 4 5 6

444 * 4 »44 *. 4,444 4,444 444 444

4 4 4 4 4 5

4 · 4 44 4 444 44 (4 4 extraction with Et ₂ O, followed by washing with aqueous K ₂ CO ₃ and aqueous HCl.

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 0.87 (d, J = 6.8 Hz, 6H), 1.01 to 1.17 (m, 2H), 1.4 (d, J = 7, 2 Hz, 3H), 1.40-1.62 (m, 4H), 1.70-1.83 (m, 2H), 1.89 (m, 1H, 2.15 (m, 3H), 3 86 (m, 2H); 4.55 (m, 1H); 6.30 (d, J = 7.1 Hz, 1H).

¹³ C-NMR (CDC1 _3): δ = 18.4, 18.78, 18.80, 24.8 (very high), 27.5, 32.2, 32.32, 36.9, 42.5 , 47.7, 71.2, 172.2, 173..

Elemental Analysis

Calcd (%): C, 65.85; H, 9.87; N, 5.49;

Found (%):, 66.01; H, 10.08; N, 5.4.

C ₁₄ H ₂₅ NO ₃ (MW = 255, 36, mass spectroscopy (MH ⁺ 256))

Example A72

Synthesis of N - [(cyclohex-1-enyl) acetyl] -L-alanine as isobutyl ester

Following the procedure of General Procedure B above, using cyclohex-1-enylacetic acid (Alpha) and L-alanine as isobutyl ester hydrochloride (from Example B above), the title compound was obtained as a solid, m.p. ° C-50 ° C. The reaction was monitored by thin layer chromatography on silica gel (R _e = 0.40 in 1: 3 EtOAc: hexane) and purification was performed by extraction with Et ₂ O followed by washing with aqueous K ₂ CO ₃ and aqueous HCl.

NMR data was as follows:

^λ H-nmr (CDCl ₃ ): δ = 0.91 (d, J = 4.5 Hz, 3H), 0.93 (d, J = 6.7 Η, 3H), 1.40 (d, J = 7.2 Hz, 3H), 1.52-1.70 (m, 4H), 1.97 (m, 3H), 2.06 (b, 2H), 2.89 (s, 2H), 3 92 (m, 2H), 4.59 (m, 1H), 5.65 (s, 1H), 6.33 (d, J = 6.6 Hz, 1H).

106

9 9 · • 9 99 • ·· • 9 9 9 9 · 9 .9 • »··· 999 9 9 9 9 • • · 9 99 9 • · · 9 9 ··

¹³ C-nmr (CDCl ₃ ): δ = 18.7, 18, 91, 18.93, 21, 9, 22.7, 25, 3, 27.6, 28.3, 46.1, 47.9 , 71.4, 127.1, 132.5, 170.6, 173.1. Elemental Analysis

Calcd (%): C, 67.38; H, 9.42; N, 5.24;

Found (%): C, 67.34; H, 9.54; N, 5.16.

C ₁₅ H ₂₅ NO ₃ (MW = 267.37, mass spectroscopy (MH ⁺ 268)).

Example A73

Synthesis of N - [(3-chlorophenyl) acetyl] alanine as 3-methylbut-2-enyl thioesters

Following the procedure of General Procedure C above and using N - [(3-chlorophenyl) acetyl] alanine and 3-methyl-2-butenedio-ester (TCI), the title compound can be prepared. The reaction was monitored by thin layer chromatography on silica gel and purification was performed by liquid chromatography using 3: 7 EtOAc: hexane as the eluent.

NMR data was as follows:

¹ H-nmr (DMSO-d ₆ ): δ = 5.2-5, 075 (m, 1H), 4.37 (dq, J = 9 Hz, 1H), 3.56 (s), 3.43 (d, J = 12Hz, 2H), 1.266 (d, J = 12Hz, 6H), 1.3 (d, J = 9Hz, 3H).

Ci6H ₂ oN0 CLS ₂ (MW = 325.86, Mass Spectroscopy (M ⁺ 325)).

Example A74

Synthesis of N - [(2-phenyl) -2-fluoroacetyl] alanine as ethyl ester

Following the procedure of General Procedure F above, using α-fluorophenyl acetic acid (Aldrich) and alanine ethyl ester (Aldrich), the title compound was prepared. The reaction was monitored by thin layer chromatography on silica gel (Rf = 0.75 in 1: 1 EtOAc / hexane) and the purification was

107 • '·· 9' 4 4

4 4 · * · ·· · · ·

44 4 -4 4 4 ·· · · · 4 4 4 · 4 4 ·

4 4 4 4 4 4 was performed by chromatography on silica gel using 1: 2 ethyl acetate / hexanes as the eluent.

NMR data was as follows:

Δ H-nmr (DMSO-d ₆ ): δ = 1.14 (q, 3H), 1.34 (d, 3H), 4.07 (m,

2H), 4.33 (m, 1H), 5.84 (d, 1H), 6.01 (d, 1H), 7.40-7.55 (m, 5H), 8.87 (m, 1H) ).

C 13 H 16 NO 3 F (MW = 253.27, mass spectroscopy (MH ⁺ 253)).

Example A75

Synthesis of N- (3,5-difluorophenylacetyl) -L-phenylglycine as methyl ester

Following the procedure of General Procedure F above and using

Of 3,5-difluorophenylacetic acid (Aldrich) and L-phenylglycine as methyl ester hydrochloride (Bachem), the title compound was prepared.

NMR data was as follows:

δ -nmr (CDCl ₃ ): δ = 7.4-7.3 (m, 5H), 6.9-6.7 (m, 3H), 6.55 (d 1H, 7.1 Hz), 5 56 (d 1H 7 Hz), 3.72 (s 3H), 3.57 (s 2H) ¹³ C-nmr (CDCl ₃ ): δ = 197.6, 177.6, 171.8, 169.3 , 136, 7, 129.6, 129.3, 127.8, 113.0, 112.9, 112.7, 111.4, 103.8, 103, 5, 65, 1, 57.2, 53 , 5, 45, 1, 43.3, 43.3

C17H15NO3F2 (MW = 319.31, mass spectroscopy (MH ⁺ 320)).

Example A76

Synthesis of N- (3,5-difluorophenylacetyl) -L-phenylglycine as isobutyl ester

3,5-difluorophenylacetic acid (Aldrich) was coupled to EDC with L-phenylglycine methyl ester hydrochloride (Bachem) using General Procedure F above. The resulting compound was placed in a large excess of the desired alcohol. A catalytic amount was added

108

• 9 · 9 9 9 . 9 9 • · · · 9 9. 9 9 9 » 9 9 .9 99 9 9 « 9. 9 • 9 9 9 9 9 9 9 9 9 9 * 9 9 9 * 9 9 9 9 9 9 9 9 9 9 9 9 '··

of anhydrous NaH and the reaction was monitored by thin layer chromatography. until the presence of the starting material has been detected. The reaction was quenched with milliliters of 1N HCl and saturated aqueous NaHCO ₃ was added after several minutes of stirring. The volume of the reaction mixture was reduced in a rotary evaporator until the excess alcohol was removed and then the remaining residue was taken up in ethyl acetate and additional water was added. The organic phase was washed with saturated aqueous NaCl and dried over MgSO ₄ . The solution was stripped of solvent on a rotary evaporator and the residue, representing the crude product, was further purified by chromatography.

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 7.35-7.3 (m, 5 m), 6.8-6.7 (m 3Η), 6.60 (d IH, 7 Hz), 5.55 (d 1Η 7.1 Hz), 3.9 (m 2H), 3.M (s 2H), 1.85 (m IH 7 Hz), 0.8 (q 6H 7 Hz) ¹³ C-nmr (CDCl 3) ₃ ): δ = 171.3, 169.3, 165.4, 138.5, 137.0,

129.5, 129.2, 127.6, 113.1, 113.0, 112.8, 112.7, 103.8,

103.5, 103.2, 75.5, 57.2, 43.4, 43.3, 28.2, 19.3

C ₂₀ H 21 NO ₃ F 2 (MW = 361.39, mass spectroscopy (MH ⁺ 362)).

Example A77

Synthesis of N- (cyclopentylacetyl) -L-phenylglycine as methyl ester

Following the general procedure D above and using cyclopentylacetic acid (Aldrich) with L-phenylglycine as the methyl ester hydrochloride (Bachem), the title compound was prepared.

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 7.35 (s, 5H), 6.44 (bd, IH), 5.6 (d, IH), 3.72 (s, 3H), 2.24 (bs, 3H), 1.9-1.4 (m, 6H), 1.2-1.05 (m, 2H)

ft · ·

109 ¹³ C-nmr (CDCl ₃ ): δ = 172.3, 171, 7, 136, 7, 129, 0, 128.6,

127.3, 56.2, 52.7, 42.5, 36.9, 32.40, 32.38, 24.8

Example A78

Synthesis of N- (cyclopentylacetyl) -L-alanine as methyl ester

Following the procedure of General Procedure D above and using cyclopentylacetic acid (Aldrich) with L-alanine as methyl ester hydrochloride (Sigma), the title compound was prepared.

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 6.38 (d, 1H), 4.50 (m, 1H), 3.65 (s, 3H), 2.13 (bs, 3H), 1.80 -1.00 (m (includes d at 1.30, 3H), 11H) ¹³ C-nmr (CDCl ₃ ): δ = 173.7, 172.5, 52.1, 47, 6, 42.3, 36, 8,

32.15, 32.14, 18.0

C11R19NO3 (MW = 213.28, mass spectroscopy (MH ⁺ 214)).

Example A79

Synthesis of N- (cyclopropylacetyl) -L-phenylglycine as methyl ester

Following the procedure of General Procedure D above and using cyclopropylacetic acid (Aldrich) with L-phenylglycine as the methyl ester hydrochloride (Bachem), the title compound was prepared.

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 7.35 (m, 5H), 6.97 (m, J = 7.2 Hz, 1H)

5.59 (d, J = 7.8Hz, 1H), 3.71 (s, 3H), 2.17 (m, 2H), 1.050.95 (m, 1H), 0.62 (m, 2H) ), 0.02 (m, 2H) ¹³ C-nmr (CDCl ₃ ): δ = 171.9, 174.6, 136, 6, 129, 0, 128.5,

127.2, 56.1, 52.7, 41.0, 6.9, 4.37, 4.33 • · ·

110 · ♦ «« «« «

9 *

4 4 4 ► 4 4 »4

949

Example A80

Synthesis of N- (cyclopropylacetyl) -L-alanine as methyl ester

Following the procedure of General Procedure D above and using cyclopropyl acetic acid (Aldrich) with L-alanine as methyl ester hydrochloride (Sigma), the title compound was prepared.

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 6.60 (d, 1H), 4.55 (m, 1H), 3.69 (with, 3H), 2.10 (m, 2H), 1.34 (d, 3H), O, 95 (m, 1H), 0.58 (m, 2H) 0.15 (m. 2H) ¹³ C-nmr (CDCl ₃ ): δ = 173.7, 172.3, 52,3, 47.7, 41.0, 18 , 2, 6.7, 4.27, 4.22

Example A81

Synthesis of N - [(3-nitrophenyl) acetyl] -L-methionine as isobutyl ester

Following the general procedure H above and using nitrophenylacetic acid (Aldrich) and L-methionine (Aldrich), the title compound was obtained as a brown oil. The reaction was monitored by thin layer chromatography on silica gel.

NMR data was as follows:

1 H-nmr (CDCl ₃ ): δ = 8.16 (m, 2H) 7.67 (d, 1H) 7.32 (t, IH), 6.31 (bd, 1H), 4.69 (m, 1H); 3.90 (d, 2H); 3, 68 (with, 2H), 2.47 (t, 2 H), 2.15 (m, 1H), 2.02 (s, 3H), 1, 90 (m, 2H), 0.91 (d, 6H). CL7H24N2O5S (MW = 368, 4, mass spectroscopy (MH ⁺ . 368)).

0 - 111 -. 0 0 0 0 0 0 0 0 • 0 0 0 0 0 0 00 0 0 000.0 ·. 0 »0 0 0 0 000 000 0 0 0 · - 0 0 000 000 «00 · 0 0 «· 0 0 Following general procedures A - B and Examples B1 - B2 illustrate alternative synthesis N- (aryl / heteroarylacetyl) amino acids, usable as

starting material for preparing the amides of the present invention.

General procedure

Preparation of acid chloride

3,5-Difluorophenylacetic acid (30 g, 0.174 mol) (Aldrich) was dissolved in dichloromethane and this solution was cooled to 0 ° C. DMF (0.5 mL, catalytic) was added, followed by dropwise addition of oxalyl chloride (18 mL, 0.20 mol) over a period of 5 minutes. The reaction mixture was stirred for 3 hours and then evaporated under reduced pressure to a residue which was placed under a high vacuum pump for 1 hour to give 3,5-difluorophenylacetyl chloride as a thick yellow oil. Other acid chlorides can be prepared in a similar manner.

General procedure

Schotten-Bauman procedure

3.5-Difluorophenylacetyl chloride (from general procedure A) was added dropwise to a 0 ° C solution of L-alanine (Aldrich) (16.7 g, 0.187 mol) in 2 N sodium hydroxide (215 mL, 0.43 mol). The reaction mixture was stirred for 1 hour at 0 ° C and then overnight at ambient temperature. The reaction mixture was diluted with water (100 mL), then extracted with ethyl acetate (3 x 150 mL). The organic layer was then washed with brine (200 mL), dried over MgSO 4 and roto evaporated under reduced pressure.

444 444

4 4 • 4 4 4 * 4 4 '4 4

4 4 4 • A; '«4

112

4 4 4 pressure on the residue. Recrystallization of the residue from ethyl acetate / hexanes yielded the desired product (34.5 g, 82% yield). Other acid chlorides may be used in this procedure to provide intermediates useful in the present invention.

Example Β 1

Synthesis of N- (phenylacetyl) -L-alanine Following the procedure of General Procedure B, the title compound prepared from (Aldrich) and L-alanine (Aldrich), m.p. 102-104 ° C.

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 9.14 (br s, 1H),

6.20 (d, J = 7.0 Hz, 1H), 4.55 (m, 1H) (d, J = 7.1 Hz, 3H).

the above was phenylacetyl chloride as a solid,

7.21-7.40 (m, 5H);

3.61 (s, 2H), 1.37 ¹³ C-nmr (CDCl ₃ ): δ = 176.0, 171.8, 134.0, 129.4, 127.5,

48.3, 43.2, 17.9.

Example B2

Synthesis of N- (3,5-Difluorophenylacetyl) -L-alanine

Following the general procedure B above, the title compound was prepared from 3,5-difluorophenylacetyl chloride (from general procedure A above) and L-alanine (Aldrich).

NMR data was as follows:

^* H-NMR (CD ₃ OD): δ = 8.32 (br s, 0.3H), 6.71 (m, 2H), 6.60 (m, IH), 4.74 (br s, 1 7H), 4.16 (m, 1H), 3.36 (s, 2H), 1.19 (d, J = 7.3 Hz, 3H).

- 113 -

99 • ·· • · • · • • · «· • · 11 ♦ • • · • 1 • · f 9 • • • 199 999 1 '. . • • • • 9 • · ·· · ♦ · · • · • ·

¹³ C-nmr (CD 3 OD): δ = 175, 9, 172, 4, 164.4 (dd, J = 13.0, 245.3 Hz), 141.1, 113.1 (dd, J = 7, 8, 17.1 Hz), 102.9 (t, J = 25.7 Hz), 49.5, 42.7, 17.5.

The compounds listed in Examples 1-22 were prepared by one of the general procedures A-G unless otherwise noted.

General procedure

EDC coupling procedure

To a 1: 1 mixture of the corresponding carboxylic acid and amino ester / amide in CH 2 Cl 2 at 0 ° C was added 1.5 equivalents of triethylamine followed by 2.0 equivalents of hydroxybenzotriazole monohydrate followed by 1.25 equivalents of 1- (3-dimethylaminopropyl) - 3-ethylcarbodiimide (EDC). The reaction mixture was stirred overnight at ambient temperature and then transferred to a separatory funnel. The mixture was washed with water, saturated aqueous NaHCO 3, 1 N aqueous hydrochloric acid, and saturated aqueous sodium chloride, and then dried over MgSO 4. The solution was stripped of solvent on a rotary evaporator to give the crude product.

General procedure

EDC coupling procedure II

The round bottom flask was charged with the corresponding carboxylic acid (1.0 equiv.), Hydroxybenzotriazole hydrate (1.1 equiv.) And the corresponding amine (1.0 equiv.) In THF under a nitrogen atmosphere. An appropriate amount (1.1 equiv for free amine and 2.2 equiv for amine hydrochloride salt) of a suitable base such as Hunig's base was added to the stirred mixture, followed by 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC) ( 1.1 equiv.). After stirring after

114 • · · · · · · · · · · · · · · · ·; · · ·

Φ Φ Φ · ·· ·· · · · · · φ φ

ΦΦΦ ··· • ·

For about 4 hours to 17 hours at ambient temperature, the solvent was removed under reduced pressure and the residue was taken up in EtOAc (or a similar solvent) / H ₂ O. The extracts were washed with saturated NaHCO ₃ , 1 N aqueous hydrochloric acid, brine and dried over Na ₂ SO ₄ . In some cases, the isolated product required further purification using standard procedures, such as chromatography and / or recrystallization.

General procedure

EDC coupling procedure III

A mixture of a suitable carboxylic acid (1 equiv.), 1-hydroxybenzotriazole (1.6 equiv.), A suitable amine (1 equiv.), N-methylmorpholine (3 equiv.) And dichloromethane (or DMF for insoluble substrates) cooled in an ice-bath. water, was stirred until a clear solution was obtained. EDC (1.3 equiv.) Was added to the reaction mixture and the cooling bath was allowed to warm to ambient temperature over 1-2 hours. The reaction mixture was then stirred overnight. The reaction mixture was then evaporated to dryness in vacuo and 20% aqueous potassium carbonate was added to the residue. The mixture was shaken vigorously and allowed to stand for several hours or overnight, if necessary, until the oily product solidified. The solidified product was then filtered off, washed thoroughly with 20% potassium carbonate, water, 10% HCl and water to give the product. Racemization was not observed using this procedure.

115 · fc fc fc fc fc fc

General procedure D

CDI coupling procedure

A solution of the appropriate acid (3.3 mmol) and 1,1'-carbodiimidazole (CDI) in 20 mL of THF was stirred for 2 hours. Amino acid ester hydrochloride (3.6 mmol) was added, followed by 1.5 mL (10.8 mmol) of triethylamine. The reaction mixture was stirred overnight and then dissolved in 100 mL diethyl ether, washed with 10% HCl three times, once

brine, 20% once carbonate potassium and once brine. The solution was dried over sulfate magnesium, filtered and evaporated that is earned product. General procedure CDI coupling reactions II Acid solution (1.08 mmol) and 1,1'-carbodiimidazole

(CDI, 0.972 mmol) in 10 mL THF was stirred for 1-2 hours. An appropriate amine (1.188 mmol) was added and the reaction mixture was stirred overnight. The whole reaction mixture was dissolved in 100 mL ethyl acetate, washed with 10% HCl (50 x 2 mL), once with brine, 20% once with potassium carbonate and once with brine. The solution was dried over magnesium sulfate, filtered and evaporated to give the product.

General procedure F

EDC coupling procedure IV

The round bottom flask was charged with a suitable carboxylic acid (1.0 equiv.), Hydroxybenzotriazole hydrate (1.1 equiv.) And an appropriate amine (1.0 equiv.) In THF under a nitrogen atmosphere. An appropriate amount (1.1 equiv for free amine and 2.2 equiv for amine hydrochloride salt) of a suitable base such as Hunig's base was added to the stirred mixture, followed by 1- (3-dimethylaminopropyl) -3 • 0 hydrochloride.

- 115 0 0 »0 0 0

I 0 ·, 0.

000 000. 0 · 0 ethylcarbodiimide (EDC) (1.1 eq). After stirring for about 4 hours to 17 hours at ambient temperature, the solvent was removed under reduced pressure and the residue was taken up in EtOAc (or a similar solvent) / H ₂ O. The extracts were washed with saturated NaHCO 3, 1 N aqueous hydrochloric acid, brine and dried over Na ₂ SO ₄ . In some cases, the isolated product required further purification using standard procedures such as chromatography and / or recrystallization.

dried (sodium sulfate) white solid, silica gel using

Example A

Synthesis of 2-amino-1-phthalimidopentane hydrochloride 2-Amino-1-pentanol was stirred in a mixture of chloroform and saturated aqueous sodium bicarbonate. Di-tert-butyl dicarbonate (1.05 equiv) was added in one portion and the mixture was stirred until starting material was consumed. The organic portion was separated, dried (sodium sulfate) and concentrated. The crude material was purified by silica gel chromatography using 1: 1 ethyl acetate / hexanes. The product was dissolved in THF. Triethylamine (1.1 equiv.) Was added and the mixture was cooled in an ice bath. Methanesulfonyl chloride (1.1 equiv.) Was added dropwise and the mixture was stirred until starting material was consumed. The mixture was concentrated under reduced pressure and then partitioned between ethyl acetate and water. The organic portion was separated, and concentrated to give which was chromatographed on 30% ethyl acetate in hexanes and finally crystallized from 1-chlorobutane / hexanes. The crystalline product was stirred in anhydrous DMF and potassium phthalimide (1.1 eq) was added. The mixture was stirred for 18 hours and then concentrated under reduced pressure

117 ··· ······························ · ··· · ♦ ·· Pressure. The residue was partitioned between ethyl acetate and water. The organic portion was dried and concentrated to give a white solid. The solid was taken up in chloroform and filtered through a pad of silica. The eluting solvent containing the product was concentrated to give the crude product as a white solid. The white solid was taken up in anhydrous dioxane and the resulting solution was saturated with HCl gas. After stirring for 30 minutes, the mixture was concentrated to give a white solid which was titrated in ether. The title compound was collected, washed with ether and dried in a vacuum oven.

Example B

Synthesis of 5-aminodibenzosuberan

5-Chlorodibenzosuberan (Aldrich) was heated to reflux in 7N NH ₃ in MeOH. After 18 hours, the reaction mixture was concentrated to a solid, which was purified by silica gel chromatography to give the title compound.

Example 1

Synthesis of N- (3-hydroxyphenyl) -N '- (phenylacetyl) -L-alaninamide Following the procedure of General Procedure C and using N-phenylacetyl) -L-alanine (Example B1) and 3-hydroxyaniline (Aldrich) the title compound was prepared as solid (m.p. = 164-167 ° C). The product was purified by extraction with EtOAc and washing with aqueous NaHCO ₃ and HCl. NMR data was as follows:

¹ H-nmr (DMSO-d ₆ ): δ = 9.90 (s, 1H), 9.40 (s, 1H), 8.41 (d, J = 7.4 Hz, IH), 7.27 (m, 4H), 7.20 (m, 2H), 7.07 (t, J =

118 ft »· ft · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·. · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·. · Ftft ftft ftftft ftftft ft · ··

8.0 Hz, 1H), 6.98 (m, 1H), 6.46 (dd, J = 1.7, 7.1 Hz, 1H), 4.43 (m, 1H), 3.50 ( s, 2H), 1.30 (d, J = 7.1Hz, 3H). ¹³ C-NMR (DMSO-d _6): δ = 171.2, 170.0, 157, 6, 140, 0, 136, 4,

129.4, 129.1, 128.2, 126.3, 110.5, 110.0, 106.4, 49.1,

41.9, 18.4.

C ₁₇ H ₁₈ N ₂ O ₃ (MW = 298.34);

mass spectrometry (MH ⁺ ) 297 (MH) ⁺ .

Example 2

Synthesis of N- (3-methoxyphenyl) -Ν '- (phenylacetyl) -L-alaninamide Following the general procedure E and using N (phenylacetyl) -L-alanine (Example B1) and 3-methoxyaniline (Aldrich), the title compound was prepared. as a solid (m.p. = 154-157 ° C).

NMR data was as follows:

δ -nmr (CDCl ₃ ): δ = 9.38 (s, 1H), 7.20-7.35 (m, 6H), 7.007.15 (m, 2H), 6.91 (d, J = 8) 2 Hz, 1H), 6.62 (dd, J = 2.2,

8.1 Hz, 1H), 4.85 (quint., J = 7.2 Hz, 1H), 3.70 (s, 3H),

3.57 (s, 2H); 1.42 (d, J = 7.0 Hz, 3H).

¹³ C-nmr (CDCl ₃ ): δ = 171, 6, 171, 0, 159, 9, 139, 1, 134.4,

129.5, 129.2, 128.8, 127.2, 112.2, 110.1, 105.5, 55.1,

49, 8, 43, 3, 18.5.

C ₁₈ H ₂ oN ₂ 0 ₃ (MW = 312.37);

mass spectroscopy (MH ⁺ ) 313.

119 ··. ·.

• · · · · · ··· ··. ·.

··························

Example 3

Synthesis of N- (3-ethoxycarbonylphenyl) -N '- (phenylacetyl) -Lalaninamide

Following general procedure E and using N (phenylacetyl) -L-alanine (Example B1) and ethyl 3-aminobenzoate (Aldrich), the title compound was prepared as a solid (m.p. = 142-145 ° C).

NMR data was as follows:

^λ H-nmr (CDCl ₃ ): δ = 9.44 (s, 1H), 8.16 (d, J = 1.8 Hz, 1H),

7.74 (dd, J = 5.0, 8.0 Hz, 2H), 7.21-7.36 (m, 6H), 6.71 (d,

J = 7.6 Hz, 1H), 4.86 (m, 1H), 4.35 (q, J = 7.1 Hz, 2H),

3.65 (s, 2H), 1.42 (d, J = 7.0 Hz, 3H), 1.35 (t, J = 7.8)

Hz, 3H).

¹³ C-nmr (CDCl ₃ ): δ = 171.5, 170, 8, 166.2, 138.2, 134.2,

131.0, 129.2, 129.0, 128.9, 127.4, 125.2, 124.2, 120.7,

61.1, 49.7, 43.5, 18.5, 14.3.

C ₂ oH ₂₂ N 20 ₄ (MW = 354.41);

mass spectroscopy (MH ⁺ ) 355.

Example 4

Synthesis of N- (4-ethoxycarbonylphenyl) -1 '- (phenylacetyl) -Lalaninamide

Following general procedure E and using N (phenylacetyl) -L-alanine (Example B1) and ethyl 4-aminobenzoate (Aldrich), the title compound was prepared as a solid (m.p. = 175-178 ° C).

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 9.61 (s, 1H), 7.91 (dd, J = 7.1, 8.9)

Hz, 2H), 7.50 (dd, J = 5.2, 8.7 Hz, 2H), 7.21-7.38 (m, SH), 6.86 (d, J = 7.6 Hz) 1 H, 4.88 (quint., J = 7.1 Hz, 1H), 4.35 (q, J = 7.1 Hz, 2H), 3.62 (s, 2H), 1.36- 1.44 (m, 6H).

120 * · ·· · · · · ·

.. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. 9 9 9 9 9 9 9 9 9

999 999

9

99 ¹³ C-nmr (CDCl ₃ ): δ = 171.7, 171.0, 166, 1, 142.1, 134.2,

130.5, 129.2, 128.9, 127.5, 125.7, 118.9, M.8, 49.8, 43.3, 18.6, 14.3.

C20H22N2O4 (MW = 354.41);

mass spectrometry (MH ⁺ ) 353 (MH) ⁺ .

Example 5

Synthesis of N- (η-Hexyl) -N '- (3,5-difluorophenylacetyl) -Lalaninamide

Following general procedure B and using N- (3,5-difluorophenylacetyl) -L-alanine (Example B2) and n-hexylamine (Aldrich), the title compound was prepared as an oil. The reaction was monitored by thin layer chromatography (Rf = 0.31 in 5% MeOH / CH 2 Cl 2) and the product was purified by flash column chromatography using 5%

MeOH / CH 2 Cl 2 as the eluent.

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 7.62 (d, 1H), 7.14 (t, 1H), 6.80-6.60 (m, 3H), 4.62 (p, 1H) 3.44 (s, 2H), 3.23-3.01 (m, 2H), 1.41 (m, 2H), 1.39 (d, 3H), 1.23 (m, 8H), 0.83 (t, 3 H). ¹³ C-nmr (CDCl ₃ ): δ = 173.0, 170.3, 165.1, 165.0, 161.8,

161.7, 139.47, 139.3, 139.2, 112.8, 112.7, 112.6, 112.5,

103.3, 103.0, 102.6, 49.5, 42.9, 40.1, 31.9, 29.8, 27.0,

23.0, 19.4, 14.5.

C17H24N2O2F2 (MW = 326.39);

mass spectroscopy (MH ⁺ ) 327.

- 121 ι · · · · ·

Example 6

Synthesis of N- (n-octyl) -Ν '- (3,5-difluorophenylacetyl) -Lalaninamide

Following general procedure B and using N- (3,5-difluorophenylacetyl) -L-alanine (Example B2) and n-octylamine (Aldrich), the title compound was prepared as an oil. The reaction was monitored by thin layer chromatography (Rf = 0.35 in 5% MeOH / CH ₂ Cl ₂ ) and the product was purified by flash column chromatography using 5%

MeOH / CH ₂ Cl ₂ as the eluent.

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 7.50 (d, IH), 7.03 (t, IH), 6.75-6.60 (m, 5H), 4.M (p, H) 3.48 (s, 2H), 3.24-3.04 (m, 2H), 1.40 (m, 2H), 1.38 (d, 3H), 1.23 (m, 10H), 0.86 (t, 3 H).

¹³ C-nmr (CDCl ₃ ): δ = 172.9, 170.2, 165.2, 165.0, 161.9,

161.7, 139.3, 139.2, 139.1, 112.8, 112.7, 112.6, 112.5,

103.4, 103.0, 102.7, 49.5, 43.0, 40.1, 32.3, 29.9, 29.7,

27.4, 23.1, 19.4, 14.6.

C ₁₉ H ₂₈ N ₂ O ₂ F ₂ (MW = 354.44);

mass spectroscopy (MH ⁺ ) 355.

Example 7

Synthesis of N- (3-methoxyphenyl) -1 '- (3,5-difluorophenylacetyl) -Lalaninamide

Following the general procedure E and using N- (3,5-difluorophenylacetyl) -L-alanine (Example B2) and 3-methoxyaniline (Aldrich), the title compound was prepared.

NMR data was as follows:

^* H-NMR (DMSO-d _6): δ = 1.30 (d, J = 7.0 Hz, 3H), 3.54 (s,

2H), 3.71 (s, 3H), 4.40 (quint, J = 7.0 Hz, 1H), 6.62 (d,

122 • 00

J = 6.7 Hz, 1H), 6.9 & 7.26 (m, 5H), 7.29 (s, 1H), 8.51 (d, J = 7.1 Hz, 1H), 10.04 ( s, 1H).

¹³ C-NMR (DMSO-d _6): δ = 18.2, 41.2, 49.2, 54.9, 101, 9 (t, J = 25.5 Hz), 104.9, 108.7 , 111.4, 112.2 (dd, J = 7.3, 17.0

Hz), 129.5, 140.1, 140.7, 159.5, 162.1 (m), 168.9, 171.3. C18H18N2O3F2 (MW = 348.35); mass spectroscopy (MH ⁺ ) 349.

Example 8 Synthesis

N- (4-ethoxycarbonylphenyl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide

Following the procedure of General Procedure E and using N- (3,5-difluorophenylacetyl) -L-alanine (Example B2) and ethyl 4-aminobenzoate (Aldrich), the title compound was prepared. NMR data was as follows:

¹ H-nmr (DMSO-d 6): δ = 1.32 (m, 6H), 3.55 (s, 2H), 4.27 (q, J = 7.0 Hz, 2H), 4.42 ( quint., J = 7.0 Hz, 1H), 7.00 (m, 2H), 7.08 (m, 1H), 7.73 (d, J = 8.5 Hz, 2H), 7.91 (d, J =

8.7 Hz, 2H), 8.57 (d, J = 6.7 Hz, 1H), 10.40 (s, 1H).

¹³ C-NMR (DMSO-d _6): δ = 14.2, 17.9, 41.1, 49.4, 60.5, 101.9 (t, J = 25.3 Hz), 112.2 (dd, J = 7.4, 16.9 Hz), 118.6,

124.3, 130.3, 140.6, 140.8, 143.3, 162.1 (dd, J = 13.1,

243.7 Hz), 165.3, 169.0, 171.9.

C20H20N2O4F2 (MW = 390.39);

mass spectroscopy (MH ⁺ ) 391

Example 9

Synthesis of N- (3-ethoxycarbonylphenyl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide

123 * «· * '.3. 4 44 4

43 4 4 «4 4 4

4 4 4 444444

4 4 4 4 4 5

44 «44 440 444 4 4

Following the general procedure E and using N- (3,5-difluorophenylacetyl) -L-alanine (Example B2) and ethyl 3-aminobenzoate (Aldrich), the title compound was prepared. NMR data was as follows:

δ -nmr (CDCl ₃ ): δ = 1.34 (t, J = 7.1 Hz, 31%), 1.47 (d, J = 6.9 Hz, 3H), 3.57 (s, 2H) ), 4.33 (q, J = 7.1 Hz, 2H), 4.85 (quint., J = 7.1 Hz, 1H), 6.66 (m, 1H), 6.78 (m, 2H), 7.16 (d, J = 7.4 Hz, 1H), 7.30 (m, 1H), 7.74 (t, J = 7.8 Hz 2H),

8.12 (d J = 1.8 Hz, 1H), 9.38 (s, 1H).

^{X 3} C-NMR (CDC1 _3): δ = 14.2, 18.6, 42.7, 49, 9, 61.2, 102.8 (t,

J = 25.1 Hz), 112.2 (dd, J 8.0, 17.1 Hz), 120.9, 124.3,

125.4, 129.0, 131.1, 137.9 138.0, 163.0 (dd, J = 12.6,

247.7 Hz), 166.2, 170.2, 171.0.

C ₂₀ H ₂₀ N ₂ O ₄ F ₂ (MW = 390.39);

mass spectroscopy (MH ⁺ ) 391.

Example 10

Synthesis of N- (3-chlorophenyl) -1 '- (3,5-difluorophenylacetyl) -L alaninamide

Following general procedure E and using N- (3,5-difluorophenylacetyl) -L-alanine (Example B2) and 3-chloroaniline (Aldrich),

NMR data was us leduj ící: ^* H-NMR (DMSO-d _6): δ = 1.31 2H), 4.38 (quint. , J = 7.1 2H), 7.33 (m, 1H) 7.46 (d, 8.57 (d, 7.0 Hz, 1H), 10.26 ^{X 3} C-nmr (DMSO-d ₆ ): δ = 18.0, Hz), 112.2 (dd, J = 7.3,

130.5, 133.1, 140.4, 140.5, C ₁₇ H ₁₅ N ₂ O ₂ F ₂ Cl (MW = 352.77);

prepared the title compound.

(d, J = 7.1 Hz, 3H), 3.55 (with, Hz, IH) 7.00 (m, 2H) 7.09 (m, J = 8.2 Hz, 1H), 7.80 (with, IH), (s, 1H). 41.2, 49 , 3, 101.9 (t, J = 25, 5

16.7 Hz), 117.6, 118.6, 123.0,

140.7, 162.2 (m), 169.0, 171.7.

• ·: O

- 124 | ft ^: · · 1 tr tr · * * ♦ * * * * * * * * * * * * * * *

mass spectroscopy (MH ⁺ ) 353.

Example 11

Synthesis of N- (3,5-dichlorophenyl) -1 '- (3,5-difluorophenylacetyl) -Lalaninamide

Following the general procedure E and using N- (3,5-difluorophenylacetyl) -L-alanine (Example B2) and 3,5-dichloroaniline (Aldrich), the title compound was prepared.

NMR data was as follows:

¹ H-nmr (DMSO-d 6): δ = 1.31 (d, J = 7.0 Hz, 3H), 3.55 (s, 2H), 4.34 (m, 1H), 7.00 ( m, 2H), 7.07 (m, 1H), 7.27 (s, 1H), 7.66 (s, 2H), 8.59 (d, J = 6.6 Hz, 1H), 10, 39 (s, 1 H). ¹³ C-nmr (DMSO-d 6): δ = 17.7, 41.1, 49.5, 101.9 (t, J = 25.5 Hz), 112.3 (dd, J = 7.7, 16.9 Hz), 117.3, 122.6, 134.1,

140.4, 140.6, 140.7, 141.2, 162.1 (dd, J = 13.4, 243.9 Hz),

169.1, 172.0.

₇ Hi4N2O C ₂ F ₂ Cl 2 (MW = 387.22);

mass spectroscopy (MH ⁺ ) 387.

Example 12

Synthesis of N- (3-cyanophenyl) -1 '- (3,5-difluorophenylacetyl) -Lalaninamide

Following the general procedure E and using N- (3,5-difluorophenylacetyl) -L-alanine (Example B2) and 3-cyanoaniline (Aldrich), the title compound was prepared. NMR data was as follows:

^* H-NMR (DMSO-d _6): δ = 1.32 (t, J = 7.1 Hz, 3H), 3.55 (s,

2H), 4.39 (quint., J = 7.0 Hz, 1H), 7.00 (m, 2H), 7.08 (m, 1H), 7.52 (m, 2H), 7.82 (m, 1H), 8.08 (s, 1H), 8.60 (d, J = 6.8 Hz, 1H), 10.42 (s, 1H).

125 «0 · · 0 00 00 · · 0. · ··. 0/0 0 .0: 0 ·: '/ -0; 0 0 0 0. # ·· -; '0'':: -z ·, ··· · 0: 0 0 · 0 ·' 0, #. ¹ »/ · '' 0 i.:'.* 0« · 000 '0 · 0, .Τ · 0 .. ··. 0. ·. '0 .0 0 • 00 00. · - 000 --- 0 0 0. 00 »0 ¹³ C-nmr (DMSO-d6): δ = 17.9, 41.1, 49.4, 101.9 (t, J = 25.4 Hz), 112.2 (dd, J = 7) , 7, 17.0 Hz), 118.7, 121.8, 123.7,

126.9, 130.3, 139.7, 140.6, 140.8, 162.1 (dd, J = 13.3,

243.6 Hz), 169.1, 172.0.

C ₁₈ H ₁₅ N ₃ O ₂ F ₂ (MW = 343, 34);

mass spectroscopy (MH +) 344.

Example 13

Synthesis of N- (phthalid-6-yl) -1 '- (3,5-difluorophenylacetyl) -Lalaninamide

Following the general procedure E and using N- (3,5-difluorophenylacetyl) -L-alanine (Example B2) and 6-aminophthalide (Aldrich), the title compound was prepared as a solid (melting point = 241-245 (decomposition)). thin layer chromatography (Rf = 0.11 in 1: 1 EtOAc / hexanes) and the product was purified by precipitation from water.

NMR data was as follows:

^* H-NMR (DMSO-d _6): δ = 1.34 (d, J = 7.1 Hz, 3H), 3.56 (s,

2H), 4.42 (m, 1H), 5.36 (s, 2H), 7.00 (d, J = 6.9Hz, 2H), 7.08 (m, 1H), 7.62 ( d, J = 8.4 Hz, 1H), 7.81 (m, 1H), 8.21 (s, 1H), 8.59 (d, J = 6.8 Hz, 1H), 10.42 ( s, 1H).

¹³ C-NMR (DMSO-d _6): δ = 18.0, 41.2, 49.3, 69, 8, 101, 9 (t, J =

25.7 Hz), 112.3 (dd, J = 7.4, 16.9 Hz), 114.2, 123.4,

125.3, 125.5, 139.7, 140.6, 141.9, 162.1 (dd, J = 13.4,

243.7 Hz), 169.1, 170.8, 171.8.

C ₁₉ H ₆ N ₂ O ₄ F ₂ (MW = 374.35);

mass spectroscopy (MH +) 375.

126 • · ► 4

9 9 •

999

9 9 9 9

9 * 9

999,999 ♦ '. ft

9 .9 9 '

Example 14

Synthesis of N - [(4-methoxycarbonylphenyl) methyl] -N '- (3,5-difluorophenylacetyl) -L-alaninamide

Following the procedure of General Procedure D and using N- (3,5-difluorophenylacetyl) -L-alanine (Example B2) and methyl-4-aminomethylbenzoate. HCl (Aldrich) was the title compound as a solid (m.p. = 191-192 ° C). The reaction was monitored by thin layer chromatography (Rf = 0.19 in 1: 1 EtOAc / hexanes) and the product was purified by precipitation from water.

NMR data was as follows:

¹ H-nmr (DMSO-d ₆ ): δ = 1.25 (d, J = 7.0 Hz, 3H), 3.53 (s,

2H), 3.84 (s, 3H), 4.30 (m, 1H), 4.35 (d, J = 6.0 Hz, 2H), 6.99 (d, J = 6.9 Hz, 2H), 7.08 (t, J = 2.1Hz, 1H), 7.35 (d, J = 8.3Hz, 2H), 7.88 (d, J = 8.2Hz, 2H) 8.43 (d, J =

7.2 Hz, 1H), 8.57 (t, 5.9 Hz, 1H).

¹³ C-NMR (DMSO-d _6): δ = 18.2, 41.3, 41.7, 48.5, 52.1, 101.9 (t, 25.4 Hz), 112.2 (dd , J = 7.5, 17.0 Hz), 127.1, 128.1,

129.2, (140, 6, 140, 8 as multiplet), 145.2, 162.1 (dd, J =

13.4, 243.7 Hz), 166.1, 168.9, 172.5.

C ₂ oH ₂ oN ₂ 0 ₄ F ₂ (MW = 390.39);

mass spectroscopy (MH ⁺ ) 391.

Example 15

Synthesis of N- (1-cyano-1-phenylmethyl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide

Following General Procedure B and using N- (3,5-difluorophenylacetyl) -L-alanine (Example B2) and 1-cyano-1-phenylmethylamine hydrochloride, the title compound was prepared as a solid (m.p. = 209-213 ° C). . The reaction was monitored by chromatography on

127 '4 44. 9 Λ · .4 ''., - '. ·. ·' ·· «'. ··« «f. 4 1 ·' - <·. The thin layer (Rf = 0.5 in 10% MeOH / CHCl3) and the product was purified by recrystallization from of 1-chlorobutane / acetonitrile.

NMR data was as follows:

^* H-NMR (DMSO-d _6): δ = 3.50-3.55 (singlets, 2H); 6.18 (d,

1H).

C ₉ H ₁₇ F2N3O ₂ (MW = 357.36);

mass spectroscopy (M ⁺ ) 357.

Example 16

Synthesis of N - [(S) -1-phenylethyl] -1 '- (3,5-difluorophenylacetyl) -Lalaninamide

Following the procedure of General Procedure B and using N- (3,5-difluorophenylacetyl) -L-alanine (Example B2) and (S) - (-) - 1-phenylethylamine (Aldrich), the title compound was prepared. The reaction was monitored by thin layer chromatography (Rf = 0.45 in 9: 1 CHCl ₃ / MeOH) and the product was purified by silica gel chromatography using 95: 5 CHCl 3 / MeOH as the eluent.

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 7.18-7.28 (m, 5H), 6.81 (m, 1H), 6.67 (m, 2H), 6.63 (m, 1H) 6.60 (m, 1H); 5.01 (m, 1H); 4.44 (m, 1H); 3.36 (m, 2H); 1.43 (d, 3H); 1.36 (m, 1H); d, 3H).

Optical rotation: [a] ₂ 3 = -103 ° (c 1, MeOH).

C19H20F2N2O2 (MW = 346.38);

Mass Spectroscopy (M ⁺ ) 346.

Example 17

Synthesis of N - [(R) -1-phenylethyl] -1 '- (3,5-difluorophenylacetyl) -Lalaninamide

Following procedure General Procedure B and using N- (3,5-difluorophenylacetyl) -L-alanine (Example B2) and (R) - (+) - 1 • φ

- 128

Φ φφ • i φ φ «·» · * φ. ‘Φ *,. φ φ.

• · φ · · · · · · φ · «· φ · φ · Phenylethylamine (Aldrich) the title compound was prepared. The reaction was monitored by thin layer chromatography (Rf = 0.35 in 9: 1 CHCl ₃ / MeOH) and the product was purified by silica gel chromatography using 96: 4 CHCl ₃ / MeOH as the eluent.

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 7.31 (m, 5H), 6.78 (m, 2H), 6.71 (m,

1H), 6.32 (m, 1H), 5.01 (m, 1H), 4.46 (m, 1H), 3.45 (m,

2H), 1.43 (d, 3H), 1.30 (d, 3H).

Optical Rotation: [α] ₂₃ - + 5.97 ° (c 1, MeOH).

C ₉ H ₂₀ F ₂ N ₂ O ₂ (MW = 346.38);

Mass Spectroscopy (M ⁺ ) 346.

Example 18

Synthesis of N- [2-methoxycarbonyl-1-phenylethyl] -N '- (3,5-difluorophenylacetyl) -L-alaninamide

Following the procedure of General Procedure B and using N- (3,5-difluorophenylacetyl) -L-alanine (Example B2) and methyl 3-amino-3-phenylpropionate hydrochloride, the title compound was prepared. The reaction was monitored by tlc _(Rf = 0.45 in 9: 1 _CHCl3 / MeOH) and the product was purified by silica gel chromatography using 95: 5 _CHCl3 / MeOH as eluent.

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 7.25 (m, 5H), 6. 89-6. 67 (m, 2H), 6.37 (m, 1H), 5.36 (m, 1H) 4.52 (m, 1H), 3.63 and 3.60 (s, 3H), 3.51 and 3.45 (s, 2H), 2.80 (m, 2H), 1.37 (t , 3H).

C ₂₁ H ₂₂ F ₂ N ₂ O ₄ (MW = 404.41);

Mass Spectroscopy (M ⁺ ) 404.

A. • <

---- · • · «---- 0 i Í:

-129 '• · ·

S: ί I «I I <<<

Example 19

Synthesis of N- [2-hydroxypyridin-3-yl] -N '- (3,5-difluorophenylacetyl) -L-alarminamide

Following the general procedure C and using N- (3,5-difluorophenylacetyl) -L-alanine (Example B2) and 3-amino-2-hydroxypyridine (prepared by Pd / G hydrogenation of 2-hydroxy-3-nitropyridine under standard conditions in EtOH / HOAc), the compound was prepared from the title.

NMR data was as follows:

^* H-NMR (DMSO-d _6): δ = 1.28 (d, J = 7 Hz, 3H), 3.55 (s, 2H), 4.51 (br q, J = 7 Hz, IH) 6.20 (t, J = 7 Hz, 1H), 6.95-7.2 (m, 4H), 8.18 (dd, J = 1.7, 7.3 Hz, 1H), 8.64 (d, J = 7.0Hz,

1H), 9.24 (s, 1H).

^{X 3} C-NMR (DMSO-d _6): δ = 17.9, 41.5, 49.7, 102.0, 102.3,

102.6, 105.6, 112.5, 112.6, 112.8, 112.9, 123.4, 128.20,

112.21, 129.2, 140.8, 160.9, 164.0, 164.2, 169.6, 172.1. C ₁₆ H ₁₅ F ₂ N ₃ O ₃ (MW = 335).

Example 20

Synthesis of N-1- (phthalimido) pent-2-yl-N '- (3,5-difluorophenylacetyl) -L-alaninamide

Following the general procedure F and using N- (3,5-difluorophenylacetyl) -L-alanine (Example B2) and 2-amino-1-phthalimidopentane hydrochloride (from Example A above), the title compound was prepared. The reaction was monitored by thin layer chromatography (Rf = 0.3 in 5% MeOH / CHCl ₃ ) and the product was purified by silica gel chromatography using 5% MeOH / CHCl ₃ as the eluent, followed by recrystallization from chlorobutane / acetonitrile.

NMR data was as follows:

130

«* ·»

^* H-NMR (DMSO-d _6): δ = 4.1 (m, 2H), 7.83 (bs, 4H). C24H25N3O4F2 (MW = 457.48); mass spectroscopy (MH ⁺ ) 457.

Example 21

Synthesis

N- [α-pyridin-2-yl-benzyl] -N '- (3,5-difluorophenylacetyl) -L-alaninamide

Following the general procedure F and using N- (3,5-difluorophenylacetyl) -L-alanine (Example B2) and α- (2-pyridyl) benzylamine (Maybridge), the title compound was prepared. The product was purified by silica gel chromatography using 5% MeOH / CHCl 3 as the eluent, followed by recrystallization from n-chlorobutane / acetonitrile. NMR data was as follows:

^* H-NMR (DMSO-d _6): δ = 6.10 (d, IH), 4.46 (m, IH).

C ₂₃ H ₂₁ F ₂ N ₃ O ₂ (MW = 409.44);

mass spectroscopy (MH ⁺ ) 409.

Example 22

Synthesis of N- [2- (methoxycarbonylmethyl) benzyl] -1 '- (3,5-difluorophenylacetyl) -L-alaninamide

2- (Aminomethyl) -benzeneacetate hydrochloride was stirred in anhydrous methanol. Thionyl chloride (1.1 equiv.) Was added dropwise and the mixture was stirred at ambient temperature overnight to give methyl 2- (aminomethyl) benzeneacetate hydrochloride as a white solid.

N (3,5-difluorophenylacetyl) -L-alanine (Example B2) in dichloromethane was coupled with methyl 2- (aminomethyl) -benzeneacetate hydrochloride using EDC, HOBT, DMA to give the title compound.

MS (M ⁺ ) 404.1 \

*

131

--- 0 · ♦

0.9 9.

0 9 '0' 0 '' • '0--990 09 • 9' 9 9 0 • 09 909

0

9 0 0

Example 23

Synthesis of N- [3- (methoxycarbonyl) benzyl] -1 '- (3,5-difluorophenylacetyl) -L-alaninamide

The title compound was prepared according to the procedure described in Example 22, but using methyl 3 (aminomethyl) benzeneate hydrochloride instead of methyl 3 (aminomethyl) benzoate hydrochloride.

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 1.410 (d, 3H), 3.56 (s, 2H).

MS (M ^& lt ^{; +} & gt ^; ) 390.

Example 24

Synthesis of N- [2- (2'-methoxycarbonylmethylphenyl) benzyl] -1 '(3,5-difluorophenylacetyl) -L-alaninamide

The title compound was prepared according to the procedure described in Example 22 but using 2- (2'-methoxycarbonylmethylphenyl) benzylamine instead of methyl 3- (aminomethyl) benzoate hydrochloride.

Example 25

Synthesis of N- [2-phenylbenzyl] -1 '- (3,5-difluorophenylacetyl) -Lalaninamide

The title compound was prepared according to the procedure described in Example 22 but using 2-phenylbenzylamine instead of methyl 3- (aminomethyl) benzoate hydrochloride.

NMR data was as follows:

^* H-NMR (CDC1 _3): δ = 1.22 (d, 3H), 3.46 (s, 2H).

MS (M ^& lt ^{; +} & gt ^; ) 408.

* · ··

I «·· · ·; ·· '♦ · i · * ··'. »9. · ’··· '.»

132 Example 26

Cell search for the detection of inhibitors of βamyloid production

The amount of compounds of formula I above was tested for their ability to inhibit the formation of β-amyloid in cells containing a Swedish mutation. This screening analysis used cells (K293 = human kidney cell lines) that were stably transfected with the amylide precursor protein 751 gene (APP751) containing the double mutation Lys ₆ 5iMet ₆ 52 to Asn ₆ 5iLeu ₆ 52 (at APP751 numbering) in a manner that was described in International Patent Application Publication No. 94 / 10569® and in Citron et al. ¹² . This mutation is commonly referred to as the Swedish mutation and cells, designated "293,751 SWE, were plated in Corning 96-well plates at 1.5-2.5 χ 10 ⁴ cells per well in Dulbecco's minimum essential medium with 10% fetal addition. bovine serum. Cell counts are important to achieve ELISA results for βamyloid in the linear area of the assay (from about 0.2 to 2.5 ng per ml).

After an overnight incubation at 37 ° C in an incubator equilibrated with 10% carbon dioxide, the media was removed and replaced with 200 μΐ of compound of formula I (drug) containing the medium per well for two hours pretreatment and the cells were incubated as above. Drug stock solutions were prepared in 100% dimethylsulfoxide so that at the final drug concentration used in the processing, the dimethylsulfoxide concentration

133

0 »0 • 0 0 0 0.0 99 0 00 9 0 0 0 9 0 · 9 9 9 00 0 00 999 ♦ 9

»W ·. · V v '>. 0 0 '. ·· »0 ·

0 9, ··· ·! · Λ ',,: 0 ..

0.0, .- 0.0 ..

did not exceed 0.5% and was in fact equal to 0.1%.

At the end of the pre-treatment period, the media was again removed and replaced with media containing fresh drug as described above and the cells were incubated for an additional two hours. After this treatment, the plates were centrifuged in a Beckman GPR at 1200 rpm for five minutes at ambient temperature to pellet cell debris from the treated medium. From each well, 100 μΐ of treated media or appropriate dilution was transferred to an ELISA plate coated with antibody 266 ¹⁴ against amino acids 13-28 of the β-amyloid peptide described in International Patent Application Publication No. 94/10569 ⁸ and stored at 4 °. C overnight. An ELISA assay employing labeled antibody 6C6 ¹⁴ against amino acids 1-16 βamyloidového peptide was performed on the following day to determine the amount formed βamyloidového peptide.

The cytotoxic effects of the compounds were measured by modification of the method described by Hansen et al., ¹³ . 25 μΐ of a stock solution (5 mg / ml) of 3, (4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) was added to cells remaining in the tissue culture to a final concentration of 1 mg / ml. ml. The cells were incubated at 37 ° C for one hour and cell activity was stopped by adding an equal volume of MTT lysis buffer (20% w / v sodium dodecyl sulfate in 50% dimethylformamide, pH 4.7). Complete extraction was achieved by shaking overnight at ambient temperature. OD 562nm was measured and the difference between ₆₅ ONM in a microplate reader, Molecular Devices UV _max as an indicator

134 • »· ·» φ '·.

• ·: ·. · '<· <

• · Φ - · · · · · · - · - ·

1 1 1 ·, · ♦ ·· ♦ 190

11 • 11

ΦΦΦ »·· • ·

11 cell viability.

The results of the β-amyloid peptide ELISA were adjusted according to the standard curve and expressed as ng / ml βamyloid peptide. For normalization to cytotoxicity, these results were divided by MTT results and expressed as a percentage of the results against the drug-free control. All results represent the mean and standard deviation of at least six replicate experiments.

Test compounds were analyzed for activity to inhibit β-amyloid peptide formation in cells using this assay. The results of this assay show that each of the compounds of the present invention that was tested reduced β-amyloid peptide formation by at least 30% compared to the control.

Example 27

In vivo inhibition of β-amyloid release and / or synthesis

This example illustrates how the compounds of the present invention can be tested for in vivo inhibition of β-amyloid release and / or synthesis. 3-4 months old PDAPP mice were used for these experiments [Games et al., (1995) Nature 373: 523-527]. Depending on which compound was tested, the compound was usually prepared at a concentration of 5 or 10 mg / ml. Because of the low solubility of the compounds, they could be administered with a variety of vehicles such as corn oil (Safeway, South San Francisco, CA); 10% EtOH in corn oil (Safeway); 2-hydroxypropyl-4-cyclodextrin (Research Biochemicals

135

9 ··. 999 · 9

9 9 ·

9 9 9

International, Natick MA); and carboxymethylcellulose (Sigma Chemical Co., St. Louis MO). Specifically, for example 141, the vehicle was carboxymethylcellulose (Sigma).

Mice were dosed subcutaneously with a 26 gauge needle and after 3 hours the animals were sacrificed with CO ₂ anesthesia and blood was collected by cardiac puncture using a 1 cc 25G 5/8 syringe / needle tuberculin coated with 0.5 M EDTA, pH 8.0. . Blood was placed in a Becton-Dickinson vacuum tube containing EDTA and centrifuged for 15 minutes at 1500 g at 5 ° C. Mouse brains were then removed and the cortex and hippocampus were dissected and placed on ice.

1. Brain test

To prepare hippocampal and cortical tissue for enzyme-linked immunosorbent assay (ELISA), each brain area was homogenized in 10 volumes of ice guanidine buffer (5.0 M guanidine-HCl, 50 mM Tris-HCl, pH 8.0) using a motorized crusher Kontes ( Fisher, Pittsburgh PA). The homogenates were gently homogenized on a rotating platform for three to four hours at ambient temperature and stored at -20 ° C by quantitative determination of β-amyloid.

Brain homogenates were diluted 1:10 with ice casein buffer [0.25% casein, phosphate buffered saline (PBS), 0.05% sodium azide, 20 µg / ml aprotinin, 5 mM EDTA, pH 8.0, 10 gg / ml of leupeptin], thereby reducing the final guanidine concentration to 0.5 M prior to centrifugation at 16,000 g for 20 minutes at 4 ° C. Β - amyloid standards (amino acids) were prepared

136 •. «·,. ·. · »Β................:::::::::::::: . ♦ · ♦ · · · · ·

1-40 or 1-42) such that the final composition was equal to 0.5 M guanidine in the presence of 0.1% bovine serum albumin (BSA).

Total β-amyloid sandwich ELISA, determining both βamyloid (amino acids 1-40) and β-amyloid (amino acids 1-42), consisted of two monoclonal antibodies (mAbs) against β-amyloid. Capture antibody, 266 ^14, is specific to amino acids 13 -28 β-amyloid. 3D6 ^15, which is specific for amino acids 1-5 of β-amyloid, is biotinylated and served as the second (indicator) antibody in the test. The 3D6 biotinylation procedure uses the manufacturer's protocol (Pierce, Rockford IL) for NHS-biotin labeling of immunoglobulins except that 100 mM sodium bicarbonate buffer, pH 8.5, was used. The 3D6 antibody does not recognize secreted amyloid precursor protein (APP) or full-length APP, but only detects β-amyloid species of the aspartic amino-terminus. The assay had a lower sensitivity limit of approximately 50 pg / ml (11 pM) and showed no cross-reactivity to endogenous mouse β-amyloid peptide at concentrations up to 1 ng / ml.

The βamyloid sandwich ELISA configuration (amino acids 1-42) uses monoclonal antibody 21F12 ¹⁵ (which recognizes amino acids 33-42 βamyloid) as the first (capture) antibody. Biotinylated 3D6 is also an indicator antibody in this assay that has a lower sensitivity limit of approximately 125 pg / ml (28 pM).

266 and 21F12 capture monoclonal antibodies were

137 were plated at 10 gg / ml in 96-well immunoassay plates (Costar, Cambidge MA) overnight at ambient temperature. Plates were then aspirated and blocked with 0.25% human serum albumin in PBS buffer for at least 1 hour at ambient temperature, then stored dried at 4 ° C until use. Plates were rehydrated with Wash Buffer (Tris-Buffered Saline, 0.05% Tween 20) prior to use. Samples and standards were added to the plates and incubated overnight at 4 ° C. Plates were washed 3 times with wash buffer between each step of the assay. Biotinylated 3D6, diluted to 0.5 µg / ml in casein incubation buffer (0.25% casein, PBS, 0.05% Tween 20, pH 7.4) was incubated in the wells for 1 hour at ambient temperature. Avidin-HRP (Vector, Burlingame CA) diluted 1: 4000 in casein incubation buffer was added to the wells for 1 hour at ambient temperature. A colorimetric substrate, Slow TMB-ELISA (Pierce, Cambridge MA) was added and allowed to react for 15 minutes, after which the enzymatic reaction was stopped by adding 2 N H 2 SO 4. The reaction product was quantified using Molecular Devices V _max (Molecular Devices, Menlo Park CA) by measuring the difference in absorption at 450 nm and 650 nm.

2. Blood test

EDTA plasma was diluted 1: 1 in a sample diluent (0.2 g / l sodium phosphate. H ₂ O (monobasic), 2.16 g / l sodium phosphate. 7H ₂ O (dibasic), 0.5 g / l thimerosal , 8.5 g / l sodium chloride, 0.5 ml TritonX-405, 6.0 g / l bovine serum albumin free of globulin and water). Samples and standards in the sample diluent were tested using a total β-amyloid assay (266 capture antibody / 3D6. *. ·. *. ·. ·. ·. ·. ·. ·. · · · · ···

- 138

indicator antibodies) described above. for the brain test except that a sample diluent was used instead of a casein diluent.

In comparison to the above description, various modifications and changes to the compositions and methods may be made by those skilled in the art. All such modifications are within the scope of the appended claims.

Claims (9)

1. A compound of formula I: X 'O wherein R ¹ is selected from the group consisting of a) an alkyl, alkenyl, alkaryl, alkcycloalkyl, aryl, cycloalkyl, cycloalkenyl, heteroaryl and heterocyclic system wherein the heteroaryl or heterocyclic group is optionally substituted with 1 to 3 substituents selected from the group consisting of alkyl, alkoxy, aryl, aryloxy, halogen, nitro, thioalkoxy and thioaryloxy groups; (b) a substituted phenyl group of formula II R ^b R ^a ' ^{r (R) m} ' RhR 'wherein R represents an alkylene containing from 1 to 8 carbon atoms, m is an integer equal to 0 or 1; 4 4 4 4 4 » - 140 R ^a and R ^a 'are independently selected from the group consisting of hydrogen, hydroxy, fluoro and methyl; R ^b and R ^b 'are independently selected from hydrogen, alkyl, alkoxy, aryl, aryloxy, cyano, cycloalkyl, halogen, heteroaryl, heteroaryloxy, heterocyclic, nitro, trihalomethyl, thioalkoxy, thioaryloxy, thioheteroaryloxy and -C (O) R ⁴ wherein R ⁴ is selected from the group consisting of alkyl, aryl, alkoxy and aryloxy; and R ^c is selected from the group consisting of hydrogen, alkyl, aryl, cyano, halogen, nitro and wherein R ^b and R ^c are fused together to form a methylenedioxy ring with a phenyl ring; and when R ^b and / or R ^b 'and / or R ^c is a fluorine atom, a chlorine atom, a bromine atom and / or a nitro group, then R ^a and / or R ^a ' may also represent a chlorine atom; and (c) 1- or 2-naphthyl substituted in the 5, 6, 7 and / or 8 position by one to four substituents selected from the group consisting of alkyl, alkoxy, halogen, cyano, nitro, trihalomethyl and thioalkoxy; R ² is selected from the group consisting of hydrogen, alkyl of from 1 to 4 carbon atoms, alkylalkoxy of from 1 to 4 carbon atoms, alkylthioalkoxy of from 1 to 4 carbon atoms; and R ³ and R ³ 'are independently selected from the group consisting of: (a) a hydrogen atom with the proviso that both R ³ and R ³ 'cannot be hydrogen; (b) alkyl with the proviso that when R ³ represents a hydrogen atom, the alkyl group R ³ 'has a straight carbon chain of at least 5 carbon atoms from the nitrogen atom, the chain optionally being substituted by one or more alkyl groups and with another condition that when both R ³ and R ³ 'are alkyl, the at least one alkyl group has a carbon chain of at least 5 carbon atoms, the chain optionally being substituted by one or more alkyl groups; • ο 9 9 9 9 9 99 99 141 (c) - (R ⁷ ) n (W) _p , wherein R ⁷ represents an alkylene group selected from the group consisting of: (i) a 999 group, W is selected from the group consisting of R ⁵ , R ⁵ ', R ⁹ and R ⁹ ' are independently selected from hydrogen, hydroxyl, alkyl, substituted alkyl, alkoxy, substituted group, alkoxy, amino , alkylamino, dialkylamino, aryl, acyl, acylamino, acyloxy, aminoacyl, cyano, cycloalkyl, halogen, carboxyl, carboxylic esters, heteroaryl and the heterocyclic system, and ^R is selected from the group consisting of hydrogen, hydroxyl, alkyl, substituted alkyl, amino, alkylamino, dialkylamino, aryl, acyl, acylamino, acyloxy, alkoxy, substituted alkoxy, aminoacyl, cyano, cycloalkyl, halogen, carboxyl, carboxylic esters, heteroaryl, heterocyclic system and wherein R ⁶ and one of R ⁵ or R ⁵ 'are fused to form a heterocyclic ring of 4 to 10 atoms having 1 to 3 heteroatoms selected from oxygen, nitrogen and a sulfur atom; with the proviso that when n is zero, R ⁹ and R ⁹ 'represent a hydrogen atom; (ii) heteroaryl; and (iii) an N-heterocyclic system, with the proviso that if W is an N-heterocyclic system, then n is not zero; and n is an integer equal to 0 or 1, and p is an integer equal to 1 to 3 with the proviso that if n is zero then p is 1, and ·· ·· «· 9 < • 9 * V ♦ · · · • • 00 (D) -CH (Φ) CH ₂ C (O) O-Q, wherein Q is selected from the group consisting of alkyl, aryl, heteroaryl, and the heterocyclic system X 'is hydrogen, hydroxy or fluoro; X represents a hydrogen atom, a hydroxy group or a fluorine atom or X 'and X together form an oxo group, Z is selected from the group consisting of a covalent bond that binds R ¹ to -CX'X-, an oxygen atom, and a sulfur atom; with the proviso that if R ¹ is phenyl, R ² is methyl, X 'and X are hydrogen, Z is a group covalently bonded to R ¹ -CX'X-, m is zero, R ³ is hydrogen , R ³ 'is - (R ⁷ ) _n (W) _p , where n is zero and _p is one and W is then (i) R ⁵ , R ⁵ ', R ⁹ , R ⁹ 'and R ⁶ are not all and (ii) R ⁵ , R ⁵ ', R ⁹ , R ⁹ ' are hydrogen and R ⁶ is methoxy; with the further proviso that when R ¹ represents 3,5-difluorophenyl, R ² represents methyl, X 'and X represent hydrogen, Z is a group that covalently binds R ¹ to -CX'X-, m is zero, R ³ represents a hydrogen atom, R ³ 'is - (R ⁷ ) _n (W) _p , wherein n is equal to one and _p is equal to one, R ⁷ represents ethylene and W represents R ⁹ 'R ⁵ then R ⁵ , R ⁵ ', R ⁹ , R ⁹ 'and R ⁶ do not all represent hydrogen; and with the further proviso that when R ¹ is 3,5-difluorophenyl, R ² is methyl, X 'and X are hydrogen, Z is a group that covalently binds R ¹ to -CX'X-am being zero, R ³ represents a hydrogen atom, R ³ 'is - (R ⁷ ) _n (W) _p , where n is zero and _p is one, W represents R ⁹ R ⁵ R ⁶ and R ⁵ ', R ⁹ , R ⁹ ' are hydrogen, then R ⁵ and R ⁶ are not condensed to form a phthalimide group with the phenyl ring to which they are attached. A compound according to claim 1, wherein R ¹ represents an unsubstituted aryl group. The compound of claim 1, wherein the unsubstituted R ¹ aryl group is selected from the group consisting of phenyl, 1-naphthyl and 2-naphthyl. • · - 144 • · · β «· · · · · · · · · · 0 9 0 9 0 0 The compound of claim 1, wherein R ¹ represents a substituted aryl group. The compound of claim 4, wherein the substituted aryl groups are substituted phenyl groups defined as follows: (a) monosubstituted phenyls having a single substitution at the 2, 3 or 4 position, each of the individual substituents being given by the corresponding groups R ^a , R ^b and R ^c ; b) disubstituted phenyls having two substituents at the positions 2.3, in positions 2,4, in positions 2,5, in positions 2,6, in positions 3.4, at the 3,5-position or at the 3,6-position, each of which is given by the corresponding groups R ^a , R ^a ', R ^b , R ^b ' and R ^c ; and (c) trisubstituted phenyls having three substituents at the 2,3,4-position, the 2,3,5-position, the 2,3,6-position, the 3,4,5-position, and the 3,4,6-position. wherein each of the substituents is given by the corresponding groups R ^a , R ^a ', R ^b , R ^b ' and R ^c . The compound of claim 5, wherein the substituted phenyl groups are selected from the group consisting of 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, 4-nitrophenyl, 4-methylphenyl, 3-methoxyphenyl, 3-nitrophenyl, 3-fluorophenyl, 3-chlorophenyl, 3-bromophenyl, 3-thiomethoxyphenyl, 3-methylphenyl, 3-trifluoromethylphenyl, 2-hydroxyphenyl, 2-methylphenyl, 2-fluorophenyl, 3,4-dichlorophenyl, 3,4-methylenedioxyphenyl, 3,5-difluorophenyl, 3,5dichlorophenyl, 2, 4-dichlorophenyl and 2,5-difluorophenyl. A compound according to claim 1, wherein R ¹ represents an alkaryl group. A compound according to claim 7, wherein R ¹ alkaryl is selected from benzyl, phenylethyl, 3-phenyl-n-propyl and 4-phenyl-n-butyl. ft · ♦ « The compound of claim 1, wherein R ¹ is selected from the group consisting of alkyl, alkenyl, cycloalkyl and cycloalkenyl groups. 10. Compound according to claim 9, ve which R ¹ represents alkyl. 11. Compound according to claim 9, ve which R ¹ represents cycloalkyl 12. Compound according to claim 9, ve which R ¹ represents alkenyl. 13. Compound cycloalkenyl. according to claim 9, ve which R ¹ represents 14. Compound according to claim 9, ve which R ¹ alkyl, cycloalkyl, alkenyl and cycloalkenyl groups are selected from sec-butyl, cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, cyclohex-1-enyl, -CH ₂ -cyclopropyl, -CH ₂ cyclobutyl, -CH ₂ -cyclohexyl, -CH _2- cyclopentyl, -CH ₂ CH ₂ -cyclopropyl, -CH ₂ CH ₂ -cyclobutyl, -CH ₂ CH ₂ -cyclohexyl and -CH ₂ CH ₂ -cyclopentyl. The compound of claim 1, wherein R ¹ is selected from the group consisting of heteroaryl and substituted heteroaryl groups. The compound of claim 15, wherein R ¹ heteroaryl and substituted heteroaryl groups are selected from pyrid-3-yl, pyrid-4-yl, thiophen-2-yl, thiophen-3-yl, benzothiazol-4- yl, 2-phenylbenzoxazol-5-yl, furan-2-yl, benzofuran-2-yl, thionaphthen-2-yl, 2-chlorothiophen-5-yl, 3-methylisoxazol-5-yl, 2- (thiophenyl) thiophen-5 -yl, 6-methoxythionaphthen-2-yl, 3-phenyl-1,2,4-thiooxadiazol-5-yl and 2-phenyl-oxazol-4-yl. A compound according to claim 1, wherein R ² is selected from the group consisting of alkyl having from 1 to 4 carbon atoms, and is selected from the group consisting of C 1 -C 4 alkyl. « Alkyl alkoxy containing from 1 to 4 carbon atoms and alkylthioalkoxy containing from 1 to 4 carbon atoms. The compound of claim 17, wherein R ² is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and -CH ₂ CH ₂ SCH ₃ . The compound of claim 1 wherein X 'and X are both hydrogen and Z is a covalent bond that binds R ¹ to CX'X-. The compound of claim 1, wherein R ³ is hydrogen and R ³ 'is selected from 3-hydroxyphenyl, 3-methoxyphenyl, 3-ethoxycarbonylphenyl, n-hexyl, n-octyl, 4-methoxycarbonylphenyl, 4-methoxycarbonylphenyl, 3- chlorophenyl, 3-cyanophenyl, 3,5-dichlorophenyl, -ΟΗ (ΟΗ ₃ ) Φ (R stereoisomer), CH (CH ₃ ) O (S stereoisomer), phthalid-6-yl, 2-hydroxypyrid-3-yl, 2 (methoxycarbonylmethyl) benzyl, 3- (methoxycarbonyl) benzyl, 2- (2'methoxycarbonylmethylphenyl) benzyl and 2-phenylbenzyl. A compound of formula I according to claim 1, selected from the group consisting of: N- (3-hydroxyphenyl) -N '- (phenylacetyl) -L-alaninamide N- (3-methoxyphenyl) -1 '- (phenylacetyl) -L-alaninamide N- (3-ethoxyphenyl) -1 '- (phenylacetyl) -L-alaninamide N- (4-ethoxycarbonylphenyl) -1 '- (phenylacetyl) -L-alaninamide N- (n-hexyl) -N '- (3,5-difluorophenylacetyl) -L-alaninamide N- (n-octyl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide • 4 4 4 · 44 ··· * 44 - 147 N- (3-methoxyphenyl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide N- (4-ethoxycarbonylphenyl) -1 '- (3,5-difluorophenylacetyl) -Lalaninamide N- (3-ethoxycarbonylphenyl) -Ν - (3,5-difluorophenylacetyl) -Lalaninamide N- (3-chlorophenyl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide N- (3,5-dichlorophenyl) -N '- (3,5-difluorophenylacetyl) -L-alaninamide N- (3-cyanophenyl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide N- (phthalid-6-yl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide N - [(4-methoxycarbonylphenyl) methyl] -1 '- (3,5-difluorophenylacetyl) -Lalaninamide N- (1-cyano-1-phenylmethyl) -N '- (3,5-difluorophenylacetyl) -Lalaninamide N - [(R) -1-phenylethyl] -N '- (3,5-difluorophenylacetyl) -L-alaninamide N - [(S) -1-phenylethyl] -N '- (3,5-difluorophenylacetyl) -L-alaninamide N- [2-hydroxypyridin-3-yl] -N '- (3,5-difluorophenylacetyl) -Lalaninamide N- [2-methoxycarbonyl-1-phenylethyl] -N '- (3,5-difluorophenylacetyl) -Lalaninamide N- [α-pyridin-2-yl-benzyl] -N '- (3,5-difluorophenylacetyl) -Lalaninamide - 148 • «· · · · N- [1- (N-phthalamido) pent-2-yl] -1 '- (3,5-difluorophenylacetyl) -Lalaninamide Ν- [2- (methoxycarbonylmethyl) benzyl] -N'- (3,5-difluorophenylacetyl) L-alaninamide N- [3- (methoxycarbonyl) benzyl] -N '- (3,5-difluorophenylacetyl) -Lalaninamide N- [2- (2'-methoxycarbonylmethylphenyl) benzyl] -N '- (3,5-difluorophenylacetyl) -L-alaninamide N- [2-phenylbenzyl] -N '- (3,5-difluorophenylacetyl) -L-alaninamide. A compound for use as a medicament for inhibiting the release of a βamyloid peptide and / or its synthesis in a cell of Formula I X '.cx R ^J Z ~ .NH C ^ NR ³ R ³ ' R ² (I) wherein R ¹ is selected from the group consisting of a) an alkyl, alkenyl, alkaryl, alkcycloalkyl, aryl, cycloalkyl, cycloalkenyl, heteroaryl and heterocyclic system wherein the heteroaryl or heterocyclic group is optionally substituted with 1 to 3 substituents selected from the group consisting of alkyl, alkoxy, aryl, aryloxy, halogen, nitro, thioalkoxy and thioaryloxy groups; «· -149 9 9999 999 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 • «· 9 (B) a substituted phenyl group of formula (II) R ^b 'R ^a ' R 2 O - (R) 2 - II R ^b R ^and wherein R represents alkylene containing from 1 to 8 carbon atoms, m is an integer equal to 0 or 1, R ^a and R ^a 'are independently selected from hydrogen, hydroxy, fluoro and methyl; R ^b and R ^b 'are independently selected from hydrogen, alkyl, alkoxy, aryl, aryloxy, cyano, cycloalkyl, halogen, heteroaryl, heteroaryloxy, heterocyclic, nitro, trihalomethyl, thioalkoxy, thioaryloxy, thioheteroaryloxy and -C (O) R ⁴ wherein R ⁴ is selected from the group consisting of alkyl, aryl, alkoxy and aryloxy; and R ^c is selected from the group consisting of hydrogen, alkyl, aryl, cyano, halogen, nitro and wherein R ^b and R ^c are fused together to form a methylenedioxy ring with a phenyl ring; and when R ^b and / or R ^b 'and / or R ^c is a fluorine atom, a chlorine atom, a bromine atom and / or a nitro group, then R ^a and / or R ^a ' may also represent a chlorine atom; and (c) 1- or 2-naphthyl substituted in the 5, 6, 7 and / or 8 position by one to four substituents selected from the group consisting of alkyl, alkoxy, halogen, cyano, nitro, trihalomethyl and thioalkoxy; R ² is selected from the group consisting of hydrogen, alkyl of from 1 to 4 carbon atoms, alkylalkoxy of from 1 to 4 carbon atoms, alkylthioalkoxy of from 1 to 4 carbon atoms; and • · · · · · · · · · · · · · · · · · · · · - 150 R ³ and R ³ 'are independently selected from the group consisting of: (a) a hydrogen atom with the proviso that both R ³ and R ³ 'cannot be hydrogen; (b) alkyl with the proviso that when R ³ represents a hydrogen atom, the alkyl group R ³ 'has a straight carbon chain of at least 5 carbon atoms from the nitrogen atom, the chain optionally being substituted by one or more alkyl groups and with another condition that when both R ³ and R ³ 'are alkyl, the at least one alkyl group has a carbon chain of at least 5 carbon atoms, the chain optionally being substituted by one or more alkyl groups; (c) - (R ⁷ ) n (W) _p , wherein R ⁷ represents an alkylene group, W is selected from the group consisting of: (and) R ⁹ R ⁵ R ⁹ 'R ⁵ wherein R ⁵ , R ⁵ ', R ⁹ and R ⁹ 'are independently selected from hydrogen, hydroxyl, alkyl, substituted alkyl, alkoxy, substituted alkoxy, amino, alkylamino, dialkylamino, aryl, acyl, acylamino, acyloxy, aminoacyl, cyano, cycloalkyl, halogen, carboxyl, carboxylic esters, heteroaryl and the heterocyclic system, and R ⁶ is selected from the group consisting of hydrogen, hydroxyl, alkyl, substituted alkyl, amino, alkylamino, dialkylamino, aryl, acyl, acylamino, acyloxy, alkoxy, substituted alkoxy, aminoacyl, cyano, cycloalkyl, halogen, carboxyl, carboxylic esters, heteroaryl, heterocyclic system and wherein R ⁶ and one of R ⁵ or R ⁵ 'are fused and 9 · Forms a heterocyclic ring of 4 to 10 atoms having 1 to 3 heteroatoms selected from the group consisting of oxygen, nitrogen and sulfur; with the proviso that when n is zero, R ⁹ and R ⁹ 'represent a hydrogen atom; (ii) heteroaryl; and (iii) an N-heterocyclic system, with the proviso that if W is an N-heterocyclic system, then n is not zero; and n is an integer equal to 0 or 1, and p is an integer equal to 1 to 3 with the proviso that if n is zero then p is 1, and (d) -CH (Φ) CH ₂ C (O) OQ wherein Q is selected from the group consisting of alkyl, aryl, heteroaryl, and the heterocyclic system X 'is hydrogen, hydroxy, or fluoro; X represents a hydrogen atom, a hydroxy group or a fluorine atom or X 'and X together form an oxo group, Z is selected from the group consisting of a covalent bond that binds R ¹ to -CX'X-, an oxygen atom, and a sulfur atom; with the proviso that if R ¹ is phenyl, R ² is methyl, X 'and X are hydrogen, Z is a group covalently bonded to R ¹ -CX'X-, m is zero, R ³ is hydrogen , R ³ 'is ~ (R ⁷ ) _n (W) _p , where n is zero and _p is one and W represents R ⁹ R ⁵ .R 6 R ⁹ 'R ⁵ then (i) R ⁵ , R ⁵ ', R ⁹ , R ⁹ 'and R ⁶ are not all hydrogen and (ii) R ⁵ , R ⁵ ', R ⁹ , R ⁹ 'are hydrogen and R ⁶ represents methoxy; ·· 4444 - 152 with the proviso that when R ¹ is 3,5-difluorophenyl, R ² is methyl, X 'and X are hydrogen, Z is a group that covalently binds R ¹ to -CX'X-, m is zero, R ³ is hydrogen, R ³ ' is - (R ⁷ ) _n (W) _p , where n is one and p is one, R ⁷ represents ethylene and W represents R ⁹ R ⁵ R ⁹ 'R ⁵ ' then R ⁵ , R ⁵ ', R ⁹ , R ⁹ ' and R ⁶ do not all represent hydrogen; and with the further proviso that when R ¹ represents 3,5-difluorophenyl, R ² represents methyl, X 'and X represent hydrogen, Z is a group that covalently binds R ¹ to -CX'X "- and m is zero, R ³ represents a hydrogen atom, R ³ 'is - (R ⁷ ) n (W) _p , where n is zero and _p is one, W represents and R ⁵ ', R ⁹ , R ⁹ 'are hydrogen, then R ⁵ and R ⁶ are not condensed to form a phthalimide group with the phenyl ring to which they are attached. 0 · 00 - 153 ···· 00 ·· * · A compound or mixture of compounds according to claim 22 for use as a medicament for preventing Alzheimer's disease in a patient at risk of Alzheimer's disease of the general formula I: X '0 r ^! of . CX. NH 4 C. nr ³ r ³ 'wherein R ¹ is selected from the group consisting of a) an alkyl, alkenyl, alkaryl, alkcycloalkyl, aryl, cycloalkyl, cycloalkenyl, heteroaryl and heterocyclic system wherein the heteroaryl or heterocyclic group is optionally substituted with 1 to 3 substituents selected from the group consisting of alkyl, alkoxy, aryl, aryloxy, halogen, nitro, thioalkoxy and thioaryloxy groups; (b) a substituted phenyl group of formula II R ^b 'R ^a II Ř ^b R ^and wherein R represents alkylene containing from 1 to 8 carbon atoms, m is an integer equal to 0 or 1, R ^a and R ^a 'are independently selected from hydrogen, hydroxy, fluoro and methyl; • · - 154 • ··· < * · · · · · R ^b and R ^b 'are independently selected from hydrogen, alkyl, alkoxy, aryl, aryloxy, cyano, cycloalkyl, halogen, heteroaryl, heteroaryloxy, heterocyclic, nitro, trihalomethyl, thioalkoxy, thioaryloxy, thioheteroaryloxy and -C (O) R ⁴ wherein R ⁴ is selected from the group consisting of alkyl, aryl, alkoxy and aryloxy; and R ^c is selected from the group consisting of hydrogen, alkyl, aryl, cyano, halogen, nitro and wherein R ^b and R ^c are fused together to form a methylenedioxy ring with a phenyl ring; and when R ^b and / or R ^b 'and / or R ^c is a fluorine atom, a chlorine atom, a bromine atom and / or a nitro group, then R ^a and / or R ^a ' may also represent a chlorine atom; and (c) 1- or 2-naphthyl substituted in the 5, 6, 7 and / or 8 position by one to four substituents selected from the group consisting of alkyl, alkoxy, halogen, cyano, nitro, trihalomethyl and thioalkoxy; R ² is selected from the group consisting of hydrogen, alkyl of from 1 to 4 carbon atoms, alkylalkoxy of from 1 to 4 carbon atoms, alkylthioalkoxy of from 1 to 4 carbon atoms; and R ³ and R ³ 'are independently selected from the group consisting of: (a) a hydrogen atom with the proviso that both R ³ and R ³ 'cannot be hydrogen; (b) alkyl with the proviso that when R ³ represents a hydrogen atom, the alkyl group R ³ 'has a straight carbon chain of at least 5 carbon atoms from the nitrogen atom, the chain optionally being substituted by one or more alkyl groups and with another condition that when both R ³ and R ³ 'are alkyl, the at least one alkyl group has a carbon chain of at least 5 carbon atoms, the chain optionally being substituted by one or more alkyl groups; (c) - (R ⁷ ) _n (W) p, wherein R ⁷ represents an alkylene group, W is selected from the group consisting of: (i) wherein R ⁵ , R ⁵ ', R ⁹ and R ⁹ ' are independently selected from hydrogen, hydroxyl, alkyl, substituted alkyl, alkoxy, substituted alkoxy, amino, alkylamino, dialkylamino, aryl, acyl, acylamino, acyloxy, aminoacyl, cyano, cycloalkyl, halogen, carboxyl, carboxylic esters, heteroaryl and the heterocyclic system, and R ⁶ is selected from the group consisting of hydrogen, hydroxyl, alkyl, substituted alkyl, amino, alkylamino, dialkylamino, aryl, acyl, acylamino, acyloxy, alkoxy, substituted alkoxy, aminoacyl, cyano, cycloalkyl, halogen, carboxyl, carboxylic esters, heteroaryl, heterocyclic system and wherein R ⁶ and one of R ⁵ or R ⁵ 'are fused to form a heterocyclic ring of 4 to 10 atoms having 1 to 3 heteroatoms selected from oxygen, nitrogen and a sulfur atom; with the proviso that when n is zero, R ⁹ and R ⁹ 'represent a hydrogen atom; (ii) heteroaryl; and (iii) an N-heterocyclic system, with the proviso that if W is an N-heterocyclic system, then n is not zero; and n is an integer equal to 0 or 1, and p is an integer equal to 1 to 3 with the proviso that if n is zero then p is 1, and (d) -CH (Φ) CH ₂ C (O) OQ wherein Q is selected from the group consisting of alkyl, aryl, heteroaryl and a heterocyclic system - 156 • 9 94 ► 9 9 9 9 9 9 494 44 99 • 9 9 9 • 9 4 4 • 9 4 • 4 9 9 ·· 49 X 'represents a hydrogen atom, a hydroxy group or a fluorine atom; X represents a hydrogen atom, a hydroxy group or a fluorine atom or X 'and X together form an oxo group, Z is selected from the group consisting of a covalent bond that binds R ¹ to -CX'X-, an oxygen atom, and a sulfur atom; with the proviso that if R ¹ is phenyl, R ² is methyl, X 'and X are hydrogen, Z is a group covalently bonded to R ¹ -CX'X-, m is zero, R ³ is hydrogen , R ³ 'is - (R ⁷ ) n (W) _p , where n is zero and _p is one and W is then (i) R ⁵ , R ⁵ ', R ⁹ , R ⁹ 'and R ⁶ are not all and (ii) R ⁵ , R ⁵ ', R ⁹ , R ⁹ ' are hydrogen and R ⁶ is methoxy; with the further proviso that when R ¹ represents 3,5-difluorophenyl, R ² represents methyl, X 'and X represent hydrogen, Z is a group that covalently binds R ¹ to -CX'X-, m is zero, R ³ represents a hydrogen atom, R ³ 'is - (R ⁷ ) _n (W) _p , wherein n is equal to one and _p is equal to one, R ⁷ represents ethylene and W represents - 157 • 4 • 4 • 4 • 4 • 44 • 4 • 44 • 44 • 44 • 4 • 4 4 • • 4 4 • · · ··· 44 R ⁹ 'R ⁵ ' then R ⁵ , R ⁵ ', R ⁹ , R ⁹ ' and R ⁶ do not all represent hydrogen; and with the further proviso that when R ¹ represents 3,5-difluorophenyl, R ² represents methyl, X 'and X represent a hydrogen atom, Z is a group that covalently binds R ¹ to -CX'X- and m is zero, R ³ represents a hydrogen atom, R ³ 'is - (R ⁷ ) _n (W) _p , where n is zero and _p is one, W represents R ⁹ R ^p R ⁶ and R ⁵ ', R ⁹ , R ⁹ ' are hydrogen, then R ⁵ and R ⁶ are not condensed to form a phthalimide group with the phenyl ring to which they are attached. 4 «* 3 • ···· - 158 »4 · 4 · 44 · 4 · 444 · 9 · 99 · 99 · 4 · 4 4 · 4 4 · 44 A compound or mixture of compounds according to claim 22 for use as a medicament for the treatment of a patient suffering from Alzheimer's disease for inhibiting a further deterioration of the condition of said patient of Formula I: X ' R‘Z .CX .NH CH nr ³ r ³ 'q wherein R is selected from the group consisting of a) an alkyl, alkenyl, alkaryl, alkcycloalkyl, aryl, cycloalkyl, cycloalkenyl, heteroaryl and heterocyclic system wherein the heteroaryl or heterocyclic group is optionally substituted with 1 to 3 substituents selected from the group consisting of alkyl, alkoxy, aryl, aryloxy, halogen, nitro, thioalkoxy and thioaryloxy groups; (b) a substituted phenyl group of formula II R ^b 'R ^a ' RHfoV (R) _m II R ^b R ^and wherein R is alkylene containing from 1 to 8 carbon atoms, • · · · - 159 - 9 is an integer equal to 0 or 1, R ^a and R ^a 'are independently selected from hydrogen, hydroxy, fluoro and methyl; R ^b and R ^b 'are independently selected from hydrogen, alkyl, alkoxy, aryl, aryloxy, cyano, cycloalkyl, halogen, heteroaryl, heteroaryloxy, heterocyclic, nitro, trihalomethyl, thioalkoxy, thioaryloxy, thioheteroaryloxy and -C (O) R ⁴ wherein R ⁴ is selected from the group consisting of alkyl, aryl, alkoxy and aryloxy; and R ^c is selected from the group consisting of hydrogen, alkyl, aryl, cyano, halogen, nitro and wherein R ^b and R ^c are fused together to form a methylenedioxy ring with a phenyl ring; and when R ^b and / or R ^b 'and / or R ^c is a fluorine atom, a chlorine atom, a bromine atom and / or a nitro group, then R ^a and / or R ^a ' may also represent a chlorine atom; and (c) 1- or 2-naphthyl substituted in the 5, 6, 7 and / or 8 position by one to four substituents selected from the group consisting of alkyl, alkoxy, halogen, cyano, nitro, trihalomethyl and thioalkoxy; R ² is selected from the group consisting of hydrogen, alkyl of from 1 to 4 carbon atoms, alkylalkoxy of from 1 to 4 carbon atoms, alkylthioalkoxy of from 1 to 4 carbon atoms; and R ³ and R ³ 'are independently selected from the group consisting of: (a) a hydrogen atom with the proviso that both R ³ and R ³ 'cannot be hydrogen; (b) alkyl with the proviso that when R ³ represents a hydrogen atom, the alkyl group R ³ 'has a straight carbon chain of at least 5 carbon atoms from the nitrogen atom, the chain optionally being substituted by one or more alkyl groups and with another condition that when both R ³ and R ³ 'are alkyl, the at least one alkyl group has a carbon chain of at least 5 carbon atoms, the chain optionally being substituted by one or more alkyl groups; • · «·· (C) - (R ⁷ ) _n (W) _p , wherein R ⁷ represents an alkylene group, W is selected from the group consisting of: (i) wherein R ⁵ , R ⁵ ', R ⁹ and R ⁹ ' are independently selected from hydrogen, hydroxyl, alkyl, substituted alkyl, alkoxy, substituted alkoxy, amino, alkylamino, dialkylamino, aryl, acyl, acylamino, acyloxy, aminoacyl, cyano, cycloalkyl, halogen, carboxyl, carboxylic esters, heteroaryl and the heterocyclic system, and R ⁶ is selected from the group consisting of hydrogen, hydroxyl, alkyl, substituted alkyl, amino, alkylamino, dialkylamino, aryl, acyl, acylamino, acyloxy, alkoxy, substituted alkoxy, aminoacyl, cyano, cycloalkyl, halogen, carboxyl, carboxylic esters, heteroaryl, heterocyclic system and wherein R ⁶ and one of R ⁵ or R ⁵ 'are fused to form a heterocyclic ring of 4 to 10 atoms having 1 to 3 heteroatoms selected from oxygen, nitrogen and a sulfur atom; with the proviso that when n is zero, R ⁹ and R ⁹ 'represent a hydrogen atom; (ii) heteroaryl; and (iii) an N-heterocyclic system, with the proviso that if W is an N-heterocyclic system, then n is not zero; and n is an integer equal to 0 or 1, and p is an integer equal to 1 to 3 with the proviso that if n is zero then p is 1, and i - 161 9 · ♦ · k t t t t t t t t t t t t t t t t t t t t (D) -CH (Φ) CH ₂ C (O) 0 -Q, wherein Q is selected from the group consisting of: alkyl, aryl, heteroaryl and heterocyclic system X 'is hydrogen, hydroxy or fluoro; X represents a hydrogen atom, a hydroxy group or a fluorine atom or X 'and X together form an oxo group, Z is selected from the group consisting of a covalent bond that binds R ¹ to -CX'X-, an oxygen atom, and a sulfur atom; with the proviso that if R ¹ is phenyl, R ² is methyl, X 'and X are hydrogen, Z is a group covalently bonded to R ¹ -CX'X-, m is zero, R ³ is hydrogen , R ³ 'is - (R ⁷ ) n (W) _p , where n is zero and _p is one and W is then (i) R ⁵ , R ⁵ ', R ⁹ , R ⁹ 'and R ⁶ are not all and (ii) R ⁵ , R ⁵ , R ⁹ , R ⁹ are hydrogen and R ⁶ is methoxy; with the further proviso that when R ¹ represents 3,5-difluorophenyl, R ² represents methyl, X 'and X represent hydrogen, Z is a group that covalently binds R ¹ to -CX'X-, m is zero, R ³ represents a hydrogen atom, R ³ 'is - (R ⁷ ) _n (W) _p , wherein n is equal to one and _p is equal to one, R ⁷ represents ethylene and W represents 0 0 00 0 0 0 0 0 0 · · 0 8 ··· 0 * 0 0 · 0 0 0 0 0 0 • · 0 0 0 0 - 161 0 »0 00 «0 0 · 0 * 0 »0 I 0 0 0 0 0 0 0 0 0 0 0 «0 R ⁹ R ⁵ O ^ _R ' R ⁹ 'R ⁵ ' then R ⁵ , R ⁵ , R ⁹ , R ⁹ and R ⁶ do not all represent hydrogen; and with the further proviso that when R ¹ represents 3,5-difluorophenyl, R ² represents methyl, X 'and X represent hydrogen, Z is a group that covalently binds R ¹ to -CX'X "- and m is zero, R ³ represents a hydrogen atom, R ³ 'is - (R ⁷ ) _n (W) _p , where n is zero and _p is one, W represents R ⁹ R ⁵ R ⁶ and R ⁵ ', R ⁹ , R ⁹ ' are hydrogen, then R ⁵ and R ⁶ are not condensed to form a phthalimide group with the phenyl ring to which they are attached. A compound according to claim 22, 23 or 24, for use as a medicament, wherein R ¹ represents an unsubstituted aryl group. The compound of claim 25, for use as a medicament, wherein the unsubstituted R ¹ aryl group is selected from the group consisting of phenyl, 1-naphthyl and 2-naphthyl. - 163 φ φ »· Φ · · φ φ φ φ φ φ φ φ Φ · φ Φ φ φ φ φ φ · · · · · Φ Φ φ φ φ »Φ Φ Φ φ φ φ φ φ β A compound according to claim 22, 23 or 24, for use as a medicament, wherein R ¹ represents a substituted aryl group. A compound according to claim 27, for use as a medicament, wherein the substituted aryl groups are substituted phenyl groups defined as follows: (a) monosubstituted phenyls having a single substitution at the 2, 3 or 4 position, each of the individual substituents being given by the corresponding groups R ^a , R ^b and R ^c ; O) disubstituted phenyls having two substituents at the positions 2.3, in positions 2,4, in positions 2,5, in positions 2,6, in positions 3.4, at the 3,5-position or at the 3,6-position, each of which is given by the corresponding groups R ^a , R ^a ', R ^b , R ^b ' and R ^c ; and (c) trisubstituted phenyls having three substituents at the 2,3,4-position, the 2,3,5-position, the 2,3,6-position, the 3,4,5-position, and the 3,4,6-position. wherein each of the substituents is given by the corresponding groups R ^a , R ^a ', R ^b , R ^b ' and R ^c . A compound according to claim 28, for use as a medicament, wherein the substituted phenyl groups are selected from the group consisting of 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, 4-nitrophenyl, 4-methylphenyl, 3-methoxyphenyl, 3-nitrophenyl, 3-fluorophenyl, 3-chlorophenyl, 3-bromophenyl, 3-thiomethoxyphenyl, 3-methylphenyl, 3-trifluoromethylphenyl, 2-hydroxyphenyl, 2-methylphenyl, 2-fluorophenyl, 3,4-dichlorophenyl, 3,4-methylenedioxyphenyl, 3,5-difluorophenyl, 3,5- dichlorophenyl, 2,4-dichlorophenyl and 2,5-difluorophenyl. A compound according to claim 22, 23 or 24, for use as a medicament, wherein R ¹ represents an alkaryl group. 0 0 9 9 Μ »ζ • 0 449 4 4 9 9 9 9 0 - 164 0 0 9 9 • · 4 4 . 4 9 4 9 9 9 0 9 9 0 0 9 44 09 A compound according to claim 30, for use as a medicament, wherein the R ¹ alkaryl group is selected from benzyl, phenylethyl, 3-phenyl-n-propyl and 4-phenyl-n-butyl. A compound according to claim 22, 23 or 24, for use as a medicament, wherein R ¹ is selected from the group consisting of alkyl, alkenyl, cycloalkyl and cycloalkenyl groups. The compound of claim represents alkyl. 32, for use as drug, where R ¹ A compound according to claim represents cycloalkyl. 32, for use as drug, where R ¹ A compound according to claim represents alkenyl. 32, for use as drug, where R ¹ A compound according to claim represents cycloalkenyl. 32, for use as drug, where R ¹ A compound according to claim 32, for use as drug, where R ¹ alkyl, cycloalkyl, alkenyl and cycloalkenyl groups are selected from sec-butyl, cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, cyclohex-1-enyl, -CH ₂ cyclopropyl, -CH ₂ -cyclobutyl, -CH ₂ -cyclohexyl, -CH ₂ -cyclopentyl, CH ₂ CH ₂ -cyclopropyl, -CH ₂ CH ₂ -cyclobutyl, -CH ₂ CH 2 -cyclohexyl and CH ₂ CH ₂ -cyclopentyl. A compound according to claim 22, 23 or 24, for use as a medicament, wherein R ¹ is selected from the group consisting of heteroaryl and substituted heteroaryl groups. A compound according to claim 38, for use as a medicament, wherein R ¹ heteroaryl and substituted heteroaryl groups are selected from pyrid-3-yl, pyrid-4-yl, thiophen-2-yl, 9 * 9 - 165 • · · β • 999 9 9 9 9 • 9 9 9 99 ·· 99 99 9 • 99 99 99 * 999 99 9 • · 9 · 999 0 9 9 ··· «· 99999 99 99 thiophen-3-yl, benzothiazol-4-yl, 2-phenylbenzoxazol-5-yl, furan-2-yl, benzofuran-2-yl, thionaphthen-2-yl, 2-chlorothiophen-5 -yl, 3-methylisoxazol-5-yl, 2- (thiophenyl) thiophen-5-yl, 6-methoxythionaphthen-2-yl, 3-phenyl-1,2,4-thiooxadiazol-5-yl and 2-phenyl-oxazol-4-yl. A compound according to claim 22, 23 or 24, for use as a medicament, wherein R ² is selected from the group consisting of alkyl of from 1 to 4 carbon atoms, alkylalkoxy of from 1 to 4 carbon atoms, and alkylthioalkoxy of from 1 to 4 carbon atoms. up to 4 carbon atoms. A compound according to claim 40, for use as a medicament, wherein R ² is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl and -CH 2 CH 2 SCH 3. A compound according to claim 22, 23 or 24, for use as a medicament, wherein X 'and X are both hydrogen and Z is a covalent bond that binds R ¹ to -CX'X-. A compound according to claim 42, for use as a medicament, wherein R ³ is hydrogen and R ³ 'is selected from 3-hydroxyphenyl, 3-methoxyphenyl, 3-ethoxycarbonylphenyl, n-hexyl, n-octyl, 4- ethoxycarbonylphenyl, 4-methoxycarbonylphenyl, 3-chlorophenyl, 3-cyanophenyl, 3,5-dichlorophenyl, -CH _(CH3) <E> (R stereoisomer) -0Η (0Η ₃₎ Φ (S stereoisomer), phthalid-6-yl, 2-hydroxypyrid-3-yl, 2- (methoxycarbonylmethyl) benzyl, 3 (methoxycarbonyl) benzyl, 2- (2'-methoxycarbonylmethylphenyl) benzyl, and 2-phenylbenzyl. A compound of formula I according to claims 22, 23 or 24, for use as a medicament selected from the group consisting of: N- (3-hydroxyphenyl) -N '- (phenylacetyl) -L-alaninamide • · ···· · · - 166 • 3 · 4 · 4 · a φ ··· · · · · 99 ·· • 9 · 9 · · · · 9 9 9 9 9 9 0 9 9 9 ft 4 4 · 0 N- (3-methoxyphenyl) -1 '- (phenylacetyl) -L-alaninamide N- (3-ethoxyphenyl) -1 '- (phenylacetyl) -L-alaninamide N- (4-ethoxycarbonylphenyl) -1 '- (phenylacetyl) -L-alaninamide N- (n-hexyl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide N- (n-octyl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide N- (3-methoxyphenyl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide N- (4-ethoxycarbonylphenyl) -1 '- (3,5-difluorophenylacetyl) -Lalaninamide N- (3-ethoxycarbonylphenyl) -1 '- (3,5-difluorophenylacetyl) -Lalaninamide N- (3-chlorophenyl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide N- (3,5-dichlorophenyl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide N- (3-cyanophenyl) -N '- (3,5-difluorophenylacetyl) -L-alaninamide N- (phthalid-6-yl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide N - [(4-Methoxycarbonylphenyl) methyl] -N '- (3,5-difluorophenylacetyl) -Lalaninamide N- (1-cyano-1-phenylmethyl) -1 '- (3,5-difluorophenylacetyl) -Lalaninamide N - [(R) -1-phenylethyl] -1 '- (3,5-difluorophenylacetyl) -L-alaninamide N - [(S) -1-phenylethyl] -N '- (3,5-difluorophenylacetyl) -L-alaninamide - 167 • 0 ·· · 0 9 99 9 99 99 • 9 9 9 9 · 9 000 000 · 0> · · 0 · 0 0 9 9 0 * 0000 0 0 0 0 0 0 0 • 00000 0 0 000 0 · 00 N- [2-Hydroxypyridin-3-yl] -1 '- (3,5-difluorophenylacetyl) -Lalaninamide N - [2-Methoxycarbonyl-1-phenylethyl] -1 '- (3,5-difluorophenylacetyl) -L alaninamide N - [α-Pyridin-2-yl-benzyl] -1 '- (3,5-difluorophenylacetyl) -Lalaninamide N- [1- (N-phthalamido) pent-2-yl] -N '- (3,5-difluorophenylacetyl) -Lalaninamide N- [2- (methoxycarbonylmethyl) benzyl] -N '- (3,5-difluorophenylacetyl) L-alaninamide N- [3- (methoxycarbonyl) benzyl] -N '- (3,5-difluorophenylacetyl) -Lalaninamide N- [2- (2'-methoxycarbonylmethylphenyl) benzyl] -Ν '- (3,5-difluorophenylacetyl) -L-alaninamide N- [2-phenylbenzyl] -1 '- (3,5-difluorophenylacetyl) -L-alaninamide. A pharmaceutical composition comprising a pharmaceutically inert carrier and a pharmaceutically effective amount of a compound of formula AND: «· • · fc« fc ·· - 168 RZZ X I .cx C' II o NH. CH NR ³ R ³ ' R ² wherein R ¹ is selected from the group consisting of a) an alkyl, alkenyl, alkaryl, alkcycloalkyl, aryl, cycloalkyl, cycloalkenyl, heteroaryl and heterocyclic system wherein the heteroaryl or heterocyclic group is optionally substituted with 1 to 3 substituents selected from the group consisting of alkyl, alkoxy, aryl, aryloxy, halogen, nitro, thioalkoxy and thioaryloxy groups; (b) a substituted phenyl group of formula (II) R ^b R ^a ' II wherein R represents alkylene containing from 1 to 8 carbon atoms, m is an integer equal to 0 or 1, R ^a and R ^a 'are independently selected from hydrogen, hydroxy, fluoro and methyl; R ^b and R ^b 'are independently selected from hydrogen, alkyl, alkoxy, aryl, aryloxy, cyano, cycloalkyl, halogen, heteroaryl, heteroaryloxy, heterocyclic, nitro, trihalomethyl, thioalkoxy, thioaryloxy, 0 0 • 0 0 0 0 0 0 0 • 0 0 0 0 0 0 0 0 - 169 0 • 0 · 0 · 0 0 0 · 0 0 0 0 0 Thioheteroaryloxy and -C (O) R ⁴ wherein R ⁴ is selected from the group consisting of alkyl, aryl, alkoxy and aryloxy; and R ^c is selected from the group consisting of hydrogen, alkyl, aryl, cyano, halogen, nitro and wherein R ^b and R ^c are fused together to form a methylenedioxy ring with a phenyl ring; and when R ^b and / or R ^b 'and / or R ^c is a fluorine atom, a chlorine atom, a bromine atom and / or a nitro group, then R ^a and / or R ^a ' may also represent a chlorine atom; and (c) 1- or 2-naphthyl substituted in the 5, 6, 7 and / or 8 position by one to four substituents selected from the group consisting of alkyl, alkoxy, halogen, cyano, nitro, trihalomethyl and thioalkoxy; R ² is selected from the group consisting of hydrogen, alkyl of from 1 to 4 carbon atoms, alkylalkoxy of from 1 to 4 carbon atoms, alkylthioalkoxy of from 1 to 4 carbon atoms; and R ³ and R ³ 'are independently selected from the group consisting of: (a) a hydrogen atom with the proviso that both R ³ and R ³ 'cannot be hydrogen; (b) alkyl with the proviso that when R ³ represents a hydrogen atom, the alkyl group R ³ 'has a straight carbon chain of at least 5 carbon atoms from the nitrogen atom, the chain optionally being substituted by one or more alkyl groups and with another condition that when both R ³ and R ³ 'are alkyl, the at least one alkyl group has a carbon chain of at least 5 carbon atoms, the chain optionally being substituted by one or more alkyl groups; (c) - (R ⁷ ) n (W) _p , wherein R ⁷ represents an alkylene group, W is selected from the group consisting of: (i) wherein R ⁵ , R ⁵ ', R ⁹ and R ⁹ ' are independently selected from hydrogen, hydroxyl, alkyl, substituted alkyl, alkoxy, substituted alkoxy, amino, alkylamino, dialkylamino, aryl, acyl, acylamino, acyloxy, aminoacyl, cyano, cycloalkyl, halogen, carboxyl, carboxylic esters, heteroaryl and the heterocyclic system, and R ⁶ is selected from the group consisting of hydrogen, hydroxyl, alkyl, substituted alkyl, amino, alkylamino, dialkylamino, aryl, acyl, acylamino, acyloxy, alkoxy, substituted alkoxy, aminoacyl, cyano, cycloalkyl, halogen, carboxyl, carboxylic esters, heteroaryl, heterocyclic system and wherein R ⁶ and one of R ⁵ or R ⁵ 'are fused to form a heterocyclic ring of 4 to 10 atoms having 1 to 3 heteroatoms selected from oxygen, nitrogen and a sulfur atom; with the proviso that when n is zero, R ⁹ and R ⁹ 'represent a hydrogen atom; (ii) heteroaryl; and (iii) an N-heterocyclic system, with the proviso that if W is an N-heterocyclic system, then n is not zero; and n is an integer equal to 0 or 1, and p is an integer equal to 1 to 3 with the proviso that if n is zero then p is 1, and (d) -CH (Φ) CH2C ( O) OQ wherein Q is selected from the group consisting of alkyl, aryl, heteroaryl and the heterocyclic system X 'is hydrogen, hydroxy or fluoro; • φ ΦΦ · 1 • 2 • · - 170 11 11 9 1 9 11 1 11 9 11 1 X represents a hydrogen atom, a hydroxy group or a fluorine atom or X 'and X together form an oxo group, Z is selected from the group consisting of a covalent bond that binds R ¹ to -CX'X-, an oxygen atom, and a sulfur atom; and with the proviso that when R ¹ is phenyl, R ² is methyl, X 'and X are hydrogen, Z is a group that covalently binds R ¹ to -CX'X-, m is zero, R ³ is hydrogen , R ³ 'is - (R ⁷ ) _n (W) _p , where n is zero and _p is one and W is then (i) R ⁵ , R ⁵ ', R ⁹ , R ⁹ 'and R ⁶ are not all and (ii) R ⁵ , R ⁵ ', R ⁹ , R ⁹ ' are hydrogen and R ⁶ is methoxy; with the further proviso that when R ¹ is 3,5-difluorophenyl, ^R2 is methyl, X 'and X are hydrogen, Z is a group that covalently links R ¹ to -CX'X "-, m is zero, R ³ represents a hydrogen atom, R ³ 'is - (R ⁷ ) _n (W) _p , wherein n is equal to one and _p is equal to one, R ⁷ represents ethylene and W represents R ⁹ 'R ⁵ then R ⁵ , R ⁵ ', R ⁹ , R ⁹ 'and R ⁶ do not all represent hydrogen; and with the further proviso that when R ¹ is 3,5-difluorophenyl, R ² is methyl, X 'and X are hydrogen, Z is a group that covalently binds R ¹ to -CX'X-am being zero, R ³ represents a hydrogen atom, R ³ 'is - (R ⁷ ) _n (W) _p , where n is zero and _p is one, W represents R ⁹ R ⁵ R ⁶ and R ⁵ ', R ⁹ , R ⁹ ' are hydrogen, then R ⁵ and R ⁶ are not condensed to form a phthalimide group with the phenyl ring to which they are attached. 45. The pharmaceutical composition according to claim 45, characterized in that R ¹ represents an unsubstituted aryl group. - 173 4 444 4 4 4 * 4 • 4 44 4 4 4 4 4 4 4 4 4 4 47. The pharmaceutical composition of claim 46 wherein the unsubstituted ^R1 aryl group is selected from the group consisting of phenyl, 1-naphthyl and 2-naphthyl. 48. A pharmaceutical composition according to claim 45, characterized in that R ¹ is substituted aryl. 49. The pharmaceutical composition of claim 48, wherein the substituted aryl groups are substituted phenyl groups defined as follows: (a) monosubstituted phenyls having a single substitution at the 2, 3 or 4 position, each of the individual substituents being given by the corresponding groups R ^a , R ^b and R ^c ; O) disubstituted phenyls having two substituents at the positions 2.3, in positions 2,4, in positions 2,5, in positions 2,6, in positions 3.4, at the 3,5-position or at the 3,6-position, each of which is given by the corresponding groups R ^a , R ^a ', R ^b , R ^b ' and R ^c ; and (c) trisubstituted phenyls having three substituents at the 2,3,4-position, the 2,3,5-position, the 2,3,6-position, the 3,4,5-position, and the 3,4,6-position. wherein each of the substituents is given by the corresponding groups R ^a , R ^a ', R ^b , R ^b ' and R ^c . 50. The pharmaceutical composition of claim 49 wherein the substituted phenyl groups are selected from the group consisting of 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, 4-nitrophenyl, 4-methylphenyl, 3-methoxyphenyl, 3-nitrophenyl, 3-fluorophenyl, 3-chlorophenyl, 3-bromophenyl, 3-thiomethoxyphenyl, 3-methylphenyl, 3-trifluoromethylphenyl, 2-hydroxyphenyl, 2-methylphenyl, 2-fluorophenyl, 3,4-dichlorophenyl, 3,4-methylenedioxyphenyl, 3,5-difluorophenyl, 3,5- dichlorophenyl, 2,4-dichlorophenyl and 2,5-difluorophenyl. 51. The pharmaceutical composition according to claim 45, characterized in that R ¹ is an alkaryl group. ·· ···· - 174 ftft ftft ftft ftft ftft ftft ftft ftft ftft ftft ftft ftft ftft ftft ft ft · Ftft ·· 52. The pharmaceutical composition of claim 51 wherein the R ¹ alkaryl group is selected from the group consisting of benzyl, phenylethyl, 3-phenyl-n-propyl and 4-phenyl-n-butyl. 53. The pharmaceutical composition according to claim 45, characterized in that R ¹ is selected from the group consisting of alkyl, alkenyl, cycloalkyl and cycloalkenyl groups. 54. The pharmaceutical composition according to claim 53, characterized in that R ¹ is alkyl. 55. The pharmaceutical composition according to claim 53, characterized in that R ¹ is cycloalkyl. 56. The pharmaceutical composition according to claim 53, characterized in that R ¹ is alkenyl. 57. The pharmaceutical composition of claim 53, wherein R ¹ is cycloalkenyl. characterized by 58. The pharmaceutical composition of claim 53 wherein said R ¹ alkyl, cycloalkyl, alkenyl and cycloalkenyl groups are selected from the group consisting of sec -butyl, cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, cyclohex-1-enyl, -CH ₂ - cyclopropyl, -CH ₂ -cyclobutyl, -CH ₂ -cyclohexyl, -CH ₂ cyclopentyl, -CH ₂ CH ₂ -cyclopropyl, -CH ₂ CH ₂ -cyclobutyl, -CH ₂ CH ₂ cyclohexyl and -CH ₂ CH ₂ -cyclopentyl. 59. The pharmaceutical composition according to claim 45, characterized in that R ¹ is selected from the group consisting of heteroaryl and substituted heteroaryl groups. 4 4 4 4 4 4 4 4 4 4 4 - 175 ·· 44 · 4 44 44 ♦ · · · φ · · 4 4 444 · 4 · 4 · 4 · 4 · 4 · 4 «44« φ 44 60. The pharmaceutical composition of claim 59 wherein R &lt; ¹ &gt; heteroaryl and substituted heteroaryl groups are selected from pyrid-3-yl, pyrid-4-yl, thiophen-2-yl, thiophen-3-yl, benzothiazol- 4-yl, 2-phenylbenzoxazol-5-yl, furan-2-yl, benzofuran-2-yl, thionaphthen-2-yl, 2-chlorothiophen-5-yl, 3-methylisoxazol-5-yl, 2- (thiophenyl) thiophene -5-yl, 6-methoxythionaphthen-2-yl, 3-phenyl-1,2,4-thiooxadiazol-5-yl and 2-phenyl-oxazol-4-yl. 61. The pharmaceutical composition according to claim 45, characterized in that R ² is selected from the group consisting of alkyl containing 1 to 4 carbon atoms, alkylalkoxy having 1 to 4 carbon atoms and alkylthioalkoxy containing from 1 to 4 carbon atoms . 62. A pharmaceutical composition according to claim 61, characterized in that R ² is selected from the group consisting of methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl and -CH2CH2SCH3. 63. The pharmaceutical composition according to claim 47, characterized in that X 'and X are both hydrogen and Z is a covalent bond, which links R ¹ to -CX'X "-. 64. The pharmaceutical composition according to claim 63, characterized in that R ³ represents a hydrogen atom and R ³ 'is selected from the group consisting of 3-hydroxyphenyl, 3-methoxyphenyl, 3ethoxykarbonylfenyl, n-hexyl, n-octyl, 4-ethoxycarbonylphenyl, 4-methoxycarbonylphenyl, 3-chlorophenyl, 3-cyanophenyl, 3,5-dichlorophenyl, -CH _(CH3) <I> (R stereoisomer), -CH _(CH3) <J> (S stereoisomer), phthalid-6-yl, 2- hydroxypyrid-3-yl, 2- (methoxycarbonylmethyl) benzyl, 3- (methoxycarbonyl) benzyl, 2- (2 ¹ methoxykarbonylmethylfenyl) benzyl, and 2-phenylbenzyl. 65. The pharmaceutical composition of claim 45 wherein the compound of formula (I) is selected from the group consisting of: N- (3-hydroxyphenyl) -N '- (phenylacetyl) -L-alaninamide N- (3-methoxyphenyl) -N '- (phenylacetyl) -L-alaninamide N- (3-ethoxyphenyl) -1 '- (phenylacetyl) -L-alaninamide N- (4-ethoxycarbonylphenyl) -1 '- (phenylacetyl) -L-alaninamide N- (n-hexyl) -N '- (3,5-difluorophenylacetyl) -L-alaninamide N- (n-octyl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide N- (3-methoxyphenyl) -N '- (3,5-difluorophenylacetyl) -L-alaninamide N- (4-ethoxycarbonylphenyl) -1 '- (3,5-difluorophenylacetyl) -Lalaninamide N- (3-ethoxycarbonylphenyl) -1 '- (3,5-difluorophenylacetyl) -Lalaninamide N- (3-chlorophenyl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide N- (3,5-dichlorophenyl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide N- (3-cyanophenyl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide N- (phthalid-6-yl) -1 '- (3,5-difluorophenylacetyl) -L-alaninamide N - [(4-methoxycarbonylphenyl) methyl] -N '- (3,5-difluorophenylacetyl) -L alaninamide - 177 · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · N- (1-Cyano-1-phenylmethyl) -1 '- (3,5-difluorophenylacetyl) -Lalaninamide N - [(R) -1-phenylethyl] -N '- (3,5-difluorophenylacetyl) -L-alaninamide N - [(S) -1-Phenylethyl] -1 '- (3,5-difluorophenylacetyl) -L-alaninamide N- [2-hydroxypyridin-3-yl] -1 '- (3,5-difluorophenylacetyl) -Lalaninamide N- [2-methoxycarbonyl-1-phenylethyl] -1 '- (3,5-difluorophenylacetyl) -L alaninamide N- [α-Pyridin-2-yl-benzyl] -1 '- (3,5-difluorophenylacetyl) -Lalaninamide N- [1- (N-phthalamido) pent-2-yl] -N '- (3,5-difluorophenylacetyl) -Lalaninamide N- [2- (methoxycarbonylmethyl) benzyl] -N '- (3,5-difluorophenylacetyl) L-alaninamide N- [3- (methoxycarbonyl) benzyl] -N '- (3,5-difluorophenylacetyl) -Lalaninamide N- [2- (2'-methoxycarbonylmethylphenyl) benzyl] -N '- (3,5-difluorophenylacetyl) -L-alaninamide N- [2-phenylbenzyl] -N '- (3,5-difluorophenylacetyl) -L-alanine d