CZ124499A3 - Esters of N-(aryl/heteroarylacetyl)amino acids, pharmaceutical compositions in which they are comprised and inhibition processes of beta-amyloid peptide release and/or synthesis thereof by making use of such compounds - Google Patents

Abstract

compounds of formula I which inhibit βamyloid release peptide and / or its synthesis. The compounds may be to use as a drug in the treatment of Alzheimer's disease, a solution to also relates to pharmaceutical compositions containing them β-amyloid peptide release inhibiting compound and / or its synthesis using such compositions.

Description

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

BACKGROUND OF THE INVENTION

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 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 N-Terminus of βAmyloid, Nature Genet., 1: 345-341 (1992).

⁸ Schenk et al., "Methods and Compositions for Detection of Soluble β-Amyloid Peptide, International Patent

4 4 · 4 9 «4 9 9 4 4 • 49 444 444

4 4 4

4 4 * 9

ApplicationPublication No. WO 94/10569. published 11 May 1994.

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

¹⁰ Losse et al., Tetrahedron, 27: 1423-1434 (1971).

¹¹ Citron et al., Mutation of the β-Amyloid Precursor Protein in Familial Alzheimer's Disease Increases β-Protein Production, Nature, 360: 672-674 (1992j.

¹² Hansen et al., "Reexamination and Further Development of a Precise and Rapid Dye Method for Measuring Cell Growth / Cell Kill, J. Immun. Meth., 119: 203-210 (1989).

¹³ P. Seubert, Nature (1992) 359: 325-327 ¹⁴ Johnson-Wood et al., PNAS USA (1997) 94: 1550-1555 ¹⁵ Tetrahedron Letters, 34 (48), 7685 (1993))

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.

/OF ·· · in »· · · »·· ·· ·· «« * ·· • · • · · · · • · '« • 9 · 9 <t · • * · · · • · · · · · Alzheimer's disease (AD) is degenerative brain disorder, characterized clinically progressive memory loss, cognitive skills, thinking, judgment and emotional stability,

which in turn 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 exhibit 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 people with Trisomy 21 (Down syndrome) and hereditary cerebral hemorrhage with tt · tt tt tt tt tt tt tt ttt tt tt tt tt · tttt · tt · tt · tttt · tt · tt · amyloidosis of the Dutch type (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 principal chemical component of amyloid plaques and vascular amyloid deposits (amyloid angiopathy) characteristic of Alzheimer's disease and other disorders listed above is a protein with a molecular weight of about 4.2 kilodaltons (kD) consisting of about 39-43 amino acids, referred to as β-amyloid peptide (βΑΡ) 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 brain.

The walls of the cerebral and meningeal blood vessels are 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.

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 diseases. Ideally, the methods of treatment would be based on agents capable of inhibiting β-amyloid peptide release and / or its synthesis in vivo.

SUMMARY 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 are therefore useful in preventing Alzheimer's disease in patients susceptible to Alzheimer's disease and / or treating Alzheimer's disease patients to inhibit further deterioration of their condition. . A class of compounds having the described properties is defined by the general formula I, which follows:

R ² in which

R ¹ is selected from the group consisting of • · ♦ · »·

9

9 9 · «·· · ·

a) alkyl, alkenyl, alkcycloalkyl, phenyl- (R) _m , -, naphthyl (R) _m -, wherein R is C 1 -C 8 alkylene and _m is an integer equal to 0 or 1, cycloalkyl, cycloalkenyl 3-pyridyl, 4-pyridyl and heteroaryl, other than 3- and 4-pyridyl, having from 3 to 10 atoms and from 1 to 4 heteroatoms selected from the group consisting of oxygen, sulfur and nitrogen, wherein the heteroaryl group is optionally substituted 1 to 3 substituents selected from the group consisting of alkyl, alkoxy, aryl, aryloxy, halogen, nitro, thioalkoxy and thioaryloxy provided that if such heteroaryl contains at least one nitrogen heteroatom, it also contains at least one oxygen and / or sulfur heteroatom;

(b) a substituted phenyl group of formula (II):

in which

R is alkylene of 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;

9 9 9 9

9 9 · 9 9

9

99,999,999

9 9

9 •

999 · · *

R ^b and R ^b 'are independently selected from hydrogen, alkyl, alkoxy, aryl, cyano, cycloalkyl, halogen, heteroaryl, 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 or 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- (R) _m - substituted at the 5-position,

6, 7 and / or 8 with one to four substituents selected from the group consisting of alkyl, alkoxy, halogen, cyano, nitro, trihalomethyl and thioalkoxy, wherein R is C 1 -C 8 alkylene and m is an integer, which is equal to 0 or 1;

R ² is selected from the group consisting of hydrogen, alkyl, phenyl, alkylalkoxy, alkylthioalkoxy; and

R ³ is selected from the group consisting of - (CH ₂ ) _n CR ¹⁰ R ⁵ R ⁶ , wherein n is an integer equal to 0, 1 or 2, R ⁵ and R ⁶ are independently selected from the group consisting of hydrogen , alkyl, alkenyl, aryl, heteroaryl, heterocyclic, -NR ⁷ R ⁸ , wherein R ⁷ and R ⁸ are each independently hydrogen or alkyl, and -COOR ⁹ , wherein R ⁹ is alkyl, and wherein further;

9 9 · 9 «9 9 9 9 9

99,999,999

9 9 9

999 99 99 ty

R ⁵ and R ⁶ may be joined to form a cycloalkyl group, a cycloalkenyl group, an aryl group, a heteroaryl group and a heterocyclic group, and if R ⁵ and R ⁶ are not joined to form an aryl or heteroaryl group, R ¹⁰ is selected from the group consisting of a hydrogen atom and an alkyl with the proviso that if n is zero then R ¹⁰ represents a hydrogen atom and if n is greater than zero and R ⁵ and R ⁶ are joined to form an aryl or heteroaryl group, then R ¹⁰ represents a bond within this group;

X represents an oxygen atom or a sulfur atom;

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, and their pharmaceutically acceptable salts, with the additional conditions that:

when R ¹ represents phenyl, R ² represents -CH (CH 3) CH ² CH 3, X represents an oxygen atom and X 1 and X represent a hydrogen atom, then R ³ does not represent -CH ₂ CH ₃ or -CH ₂ CH (CH ₃ ) ₂ ;

when R ¹ is phenyl, R ³ is -CH 2 CH (CH 3) 2, X is oxygen and X 1 and X are hydrogen, then R ^{2 is} not -CH (CH 3) ₂ when R ¹ is pyrid-3-yl, R ² is ethyl, X is oxygen and X 1 and X are hydrogen, then R ^{3 is} not -CH ₂ CH (CH ₃ ) ₂ when R ¹ is indoxazin-3-yl, 2,4-dimethylthiazol-5-yl, 4-methyl-l, 2,5- thiooxadizol-3-yl or 3,5 (trifluoromethyl) phenyl, R ² is methyl, X is oxygen, and X 'and X represent a hydrogen fy #

ft · • • ftft • ft · • · • ft • ft • • ftft · • ft ♦ • · • * • ftft ftftft • • • · • • · ftft ·· ftftft • ftft ft ft ftft

hydrogen, then R ^{3 is} not -CH 2 CH (CH 3) 2, and when R ¹ is -CH 2 -phenyl, R ³ is -CH ₂ CH ₃ , X is oxygen and X 1 and X are hydrogen, then R ^{2 is} not - CH ₂ CH (CH ₃ ) ₂ .

Surprisingly, substituents at the 2-and / or 6-position of the phenyl group are limited to the substituents listed above and larger substituents, other than those specifically specified above, eliminate the ability of the resulting compound to inhibit β-amyloid peptide release and / or synthesis .

Accordingly, in one of its method objects, the present invention relates to a method for inhibiting the release of β-amyloid peptide and / or its synthesis in a cell, the method comprising administering to the cell an amount of a compound or mixture of compounds of formula I above effective for inhibiting cellular release and / or synthesis of a β-amyloid peptide.

Since in vivo β-amyloid peptide formation is associated with the pathogenesis of Alzheimer's disease ^8.9 , compounds of Formula I may also be used in conjunction with pharmaceutical compositions for the prophylactic and / or therapeutic prevention and / or treatment of Alzheimer's disease. Therefore, in another of its subjects, 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 to said patient a pharmaceutical composition comprising a pharmaceutically inert / carrier and an effective amount of a compound or mixture of compounds of Formula 1 above.

In a further 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's condition, 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 the general 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? Governed by a corresponding group R ^a / R ^a ', R ^b / R ^b ' and R ^c ; disubstituted phenyls which include phenyls having substituents at the 2,3-position, the 2,4-position, the 2,5-position, the 2,6-position, the 3,4-position, the 3,5-position, or the 3-position, 6, wherein each of the substituents is ε governed 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 the 2,3,4-position, 2,3,5-position, 2,3,6-position, 3,4,5-position, and 3,4-position, 6, wherein each of the substituents is governed by the corresponding groups R ^a , R ^a ', R ^b , R ^b and R ^c . Preferably, substituted phenyl groups / β do not include more than 3 substituents.

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

Preferred R ¹ alkyl, alkcycloalkyl, cycloalkyl and cycloalkenyl groups include, by way of example, 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, -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, thien-2-yl, thien-3-yl, benzothiazol-4-yl, 2-phenylbenzoxazol-5-yl, furan 2-yl, benzofuran-2-yl, benzothiophen-3-yl, 2-chlorothien-5-yl, 3-methylisoxazol-5-yl, 2- (phenylthio) thien-5-yl, 6-methoxythiophen-2-yl, 3-Phenyl-1,2,4-thiooxadiazol-5-yl, 2-phenyloxazol-4-yl, and • φ · φ · • · φ · · · ·

Podobně Β Β Β podobně • ΒΒΒΒ ΒΒ Β · like.

Preferably, R ^{2 is} selected from the group consisting of alkyl having from 1 to 4 carbon atoms, phenyl, an alkyl alkoxy group containing from 1 to 4 carbon atoms, and an alkylthioalkoxy group containing from 1 to 4 carbon atoms. Particularly preferred R ² substituents include, by way of example, methyl, ethyl, n-propyl, iso-propyl, n-butyl, isobutyl, -CH2CH2SCH3, cyclohexyl and phenyl.

When X represents an oxygen atom, preferred R ³ substituents include, for example, methyl, ethyl, isopropyl, n-propyl, n-butyl, iso-butyl, cyclopentyl, allyl, iso-but-2-enyl, 3-methylpentyl, -CH 2 -cyclopropyl, CH ₂ -cyclohexyl, -CH ₂ - (3-tetrahydrofuranyl), -CH ₂ -thien-2-yl, -CH ₂ (1-methyl) cyclopropyl, -CH ₂ -thien-3-yl, -CH ₂ -C (O) Ot -butyl, CH ₂ -C (CH ₃ ) ₃ , -CH ₂ CH (CH ₂ CH ₃ ) ₂ , -2-methylcyclopentyl, cyclohex-2-enyl, -CH [CH (CH ₃ ) ₂ ] COOCH ₃ , - CH ₂ CH ₂ N (CH ₃ ) ₂ ,

CH ₂ C (CH ₃ ) = CH ₂ , -CH ₂ CH = C (CH ₃ ) _2, and the like.

When X represents a sulfur atom, preferred substituents R ³ include, for example, iso-but-2-enyl and iso-butyl.

The present invention also relates to novel pharmaceutical compositions comprising a pharmaceutically inert carrier and a compound of Formula I above.

Particularly preferred compounds for use in the methods and compositions of the present invention include, by way of example, the following compounds wherein the stereochemistry φ φ φ * φ φ • φ φ

φ φφφφ of group R ² (if applicable) is preferably derived from L-amino acid:

N- (phenylacetyl) alanine, iso-butyl ester

N- (3-phenylpropionyl) alanine, iso-butyl ester

N- (3-methylpentanoyl) alanine, iso-butyl ester

N - [(4-chlorophenyl) acetyl] alanine, isobutyl ester

N - [(3,4-dichlorophenyl) acetyl] alanine, iso-butyl ester

N - [(3-pyridyl) acetyl] alanine, isobutyl ester

N - [(1-naphthyl) acetyl] alanine, iso-butyl ester

N - [(2-naphthyl) acetyl] alanine, iso-butyl ester

N- (4-phenylbutanoyl) alanine, iso-butyl ester

N- (5-phenylpentanoyl) alanine, iso-butyl ester

N - [(4-pyridyl) acetyl] alanine, isobutyl ester

2 - [(3,4-dichlorophenyl) acetamido] butyric acid, iso-butyl ester

2 - [(3-methoxyphenyl) acetamido] butyric acid, iso-butyl ester

2 - [(4-nitrophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(3,4-methylenedioxyphenyl) acetamido] butyric acid, iso-butyl ester

2 - [(thien-3-yl) acetamido] butyric acid, iso-butyl ester 2 - [(4-chlorophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(3-nitrophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(2-hydroxyphenyl) acetamido] butyric acid, iso-butyl ester

2 - [(2-naphthyl) acetamido] butyric acid, iso-butyl ester 2 - [(2,4-dichlorophenyl) acetamido] butyric acid, iso-butyl ester

2 - [(4-bromophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(3-chlorophenyl) acetamido] butyric acid, iso-butyl ester • · «·· ··

9 · · ·

9 9 9 9 9 9 99 9

9

999 99

2 - [(3-fluorophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(benzothiazol-4-yl) acetamido] butyric acid, iso-butyl ester

2 - [(2-methylphenyl) acetamido] butyric acid, iso-butyl ester

2 - [(2-fluorophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(4-fluorophenyl) acetamido] butyric acid, 2 - [(3-bromophenyl) acetamido] butyrate, iso-butyl ester 2- [(3-trifluoromethylphenyl) acetamido] butyric acid, iso-butyl ester

2 - [(2-thienyl) acetamido] butyric acid, 2- (phenylacetamido) butyric acid iso-butyl ester, N- (phenylacetyl) valine iso-butyl ester, 2-methylbutyl ester

N- (phenylacetyl) methionine, iso-butyl ester

N- (phenylacetyl) leucine, iso-butyl ester

N - [(3-chlorophenyl) acetyl] alanine, N - [(3-chlorophenyl) acetyl] alanine, N - [(3-chlorophenyl) acetyl] alanine, N - [(3-chlorophenyl) acetyl] alanine, 2-thienylmethyl ester N- [ (3-chlorophenyl) acetyl] alanine, (1-methylcyclopropyl) methyl ester

N - [(3-chlorophenyl) acetyl] alanine, 3-thienylmethyl ester N - [(3-chlorophenyl) acetyl] alanine, 2-methylcyclopentyl ester N - [(3-chlorophenyl) acetyl] alanine, 2-methylprop-2-enyl ester N N - [(2-phenylbenzoxazol-5-yl) acetyl] alanine, N - [(3-methylthiophenyl) acetyl] alanine, isobutyl ester, - [(3-chlorophenyl) acetyl] alanine, cyclohex-2-enyl ester N-4 - [(2-furyl) acetyl] alanine, iso-butyl ester

N - [(benzofuran-2-yl) acetyl] alanine, iso-butyl ester

N - [(benzothiophen-3-yl) acetyl] alanine, iso-butyl ester N - [(2-chloro-5-thienyl) acetyl] alanine, iso-butyl ester

·· ·· · «1 2 3 4

4 4 4 4 «44 444« 4 «

4 4

Ν - [(3-methyl-isoxazol-5-yl) acetyl] alanine, N - [(2-phenylthiothienyl) acetyl] alanine iso-butyl ester, N - [(6-methoxybenzothiophen-2-yl) acetyl] isobutyl ester alanine, isobutyl ester

N - [(3-phenyl-1,2,4-thiadiazol-5-yl) acetyl] alanine, iso-butyl ester

N - [(2-Phenyloxazol-4-yl) acetyl] alanine, iso-butyl ester

N - [(3-methylphenyl) acetyl] alanine, iso-butyl ester

N - [(2,5-difluorophenyl) acetyl] alanine, iso-butyl ester

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

N - [(3-thienyl) acetyl] alanine, iso-butyl ester

N - [(4-methylphenyl) acetyl] alanine, isobutyl ester

N- (phenylacetyl) alanine, (1-methoxycarbonyl) isobutyl ester

N - [(3-nitrophenyl) acetyl] alanine, iso-butyl ester

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

N - [(3-nitrophenyl) acetyl] methionine, ethyl ester

N - [(3-chlorophenyl) acetyl] alanine, isobutyl ester

N - [(3-chlorophenyl) acetyl] alanine,

2- (N, N-dimethylamino) ethyl ester

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

N - [(3,5-dichlorophenyl) acetyl] alanine, iso-butyl ester

N- (cyclohexylacetyl) alanine, iso-butyl ester

N- (cyclopentylacetyl) alanine, iso-butyl ester

N - [(cyclohex-1-enyl) acetyl] alanine, iso-butyl ester

N - [(3-chlorophenyl) acetyl] alanine, 3-methylbut-2-enylthioester

N - [(2-phenyl) -2-fluoroacetyl] alanine, ethyl ester

N- (3,5-difluorophenylacetyl) phenylglycine methyl ester

N- (3,5-difluorophenylacetyl) phenylglycine, iso-butyl ester

N- (cyclopentylacetyl) phenylglycine, methyl ester 9

N- (cyclopentylacetyl) alanine, N- (cyclopropylacetyl) phenylglycine methyl ester, N- (cyclopropylacetyl) alanine methyl ester, N - [(3-nitrophenyl) acetyl] methionine methyl ester, iso-butyl ester

In addition, the present invention of formula III:

further relates to compounds

in which

R ¹ is selected from the group consisting of

a) alkyl, alkenyl, alkcycloalkyl, phenyl- (R) _m , -, naphthyl (R) _m -, wherein R is C 1 -C 8 alkylene and _m is an integer equal to 0 or 1, cycloalkyl, cycloalkenyl 3-pyridyl, 4-pyridyl and heteroaryl, other than 3- and 4-pyridyl, having from 3 to 10 atoms and from 1 to 4 heteroatoms selected from the group consisting of oxygen, sulfur and nitrogen, wherein the heteroaryl group is optionally substituted 1 to 3 substituents selected from the group consisting of alkyl, alkoxy, aryl, aryloxy, halogen, nitro, thioalkoxy and thioaryloxy, provided that if such heteroaryl contains at least one nitrogen heteroatom, then also it comprises at least one oxygen and / or sulfur heteroatom;

(b) a substituted phenyl group of formula (II):

(R) _n

II in which

R is C 1 -C 8 alkylene is an integer equal to 0 or 1 m

R ^{a is} selected from hydroxy, atom selected from alkyl, alkoxy, aryl, and R ^a are independently hydrogen, fluoro and methyl;

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

R ^c is selected from the group consisting of hydrogen, alkyl, aryl, cyano, halogen, nitro or 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- (R) _m -, wherein R is a C 1 -C 8 alkylene group and _m is an integer equal to 0 or 1, substituted at the 5, 6, 7 position and / or or with 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, phenyl, alkylalkoxy, alkylthioalkoxy; and

R ³ is selected from the group consisting of - (CH ₂ ) _n CR ¹⁰ R ⁵ R ⁶ , wherein n is an integer equal to 0, 1 or 2, R ⁵ and R ⁶ are independently selected from the group consisting of hydrogen , alkyl, alkenyl, aryl, heteroaryl, heterocyclic, -NR ⁷ R ⁸ wherein R ⁷ and R ⁸ are independently hydrogen or alkyl and -COOR ⁹ wherein R ⁹ is alkyl and wherein R ⁵ and R ^{6 are} furthermore they may be linked to form a cycloalkyl, cycloalkenyl, aryl, heteroaryl and heterocyclic group, and if R ⁵ and R ⁶ are not joined to form an aryl or heteroaryl group, R ¹⁰ is selected from the group consisting of hydrogen and alkyl; with the proviso that if n is zero then R ¹⁰ represents a hydrogen atom and if n is greater than zero and R ⁵ and R ⁶ are joined to form an aryl or heteroaryl group, then R ¹⁰ represents a bond within that group;

X represents an oxygen atom or a sulfur atom;

X 'is hydrogen, hydroxy or • O

0000 fluorine;

X represents a hydrogen atom, a hydroxy group or a fluorine atom or X 'and X together form an oxo group, and their pharmaceutically acceptable salts, with the additional conditions that:

when R ¹ represents phenyl, R ² represents -CH (CH 3) CH ² CH 3, X represents an oxygen atom and X 1 and X represent a hydrogen atom, then R ³ does not represent -CH ₂ CH 3 or -CH ₂ CH (CH ₃ ) ₂ ;

when R ¹ is phenyl, R ³ is -CH 2 CH (CH 3) 2, X is oxygen and X 1 and X are hydrogen, then R ^{2 is} not -CH (CH 3) 2; when R ¹ is pyrid-3-yl, R ² represents ethyl, X represents oxygen and X 'and X represent hydrogen, then R ^{3 is} not -CH ₂ CH (CH ₃ ) ₂ when R ¹ is indoxazin-3-yl, 2,4-dimethylthiazol-5-yl, 4 -methyl-1,2,5-thiooxadizol-3-yl or 3,5di (trifluoromethyl) phenyl, R ² is methyl, X is oxygen and X 'and X are hydrogen, then R ^{3 is} not -CH ₂ CH ( CH ₃ ) ₂ , and when R ¹ represents -CH 2 -phenyl, R ³ represents -CH ₂ CH ₃ , X represents an oxygen atom and X 'and X represent a hydrogen atom, then R ² does not represent -CH ₂ CH (CH ₃ ) ₂ , and in addition, with the further proviso that the following known compound is excluded:

N- (phenylacetyl) methionine ethyl ester.

Preferred compounds of formula III above include those described in formula IV, which is as follows:

· «* Φ φ φ φ φ φ φ φ φ φ φ φ * * * • * * * φ φ φ • φ • • φ • φ

R ¹ R ² R ³ X Γ - ........ ^{.. 1} -φ -CHj -CH _; CH (CCI), 0 -CtL-φ -CH, -CH., CH (CH 2), 0 -CHj -CH, CH (CH3) ₂ 1 0 4-ci- <a- -ch ₃ -CH ₂ CH (CH ₃ ) ₂ O 3.4-α-ψ- -CH, -CHjCHfCH,), 0 pyrid-3-yl -ch ₃ . -CH ₂ CH _(CH?) J 0 1-Naphthyl -CHj -CH, CH (CH3) ₂ 0 2-Naphthyl -CHj -CH ₂ CH (CH 3) ₂ 0 - (CH 2), - 0 -CHj -CH ₂ CH (CH 3) ₂ 0 - (CH ₂ ) ₃ - <p -CHj -CH, CH (CH3) i 0 pyrid-4-yl -CHj -CH ₂ CH (CH 3) ₂ 0 3.4-di-Cl-4 -CHjCH, -CH ₂ CH (CH 3), 0 3-CH3CM> - -CILCHj -CH ₂ CH (CH ₂ ) ₂ 0 4-ΝΟ ₂ -φ- -CH, CH, -CH ₂ CH (CH ₂ ) ₂ 0 3,4-methylene- dioxyphenyl- -CH, CH3 -CH ₂ CH (CH 3) ₂ 0

Tt · tttt tttt ttt t · t · t · t · t · t · t ···· * * Λ 99 9 9 9 <9 ··

R ¹ R ² R ³ X thien-3-yl -CH, CH, -CH3CH (CH3) j 0 4-C1-0- -CHjCH, -CH ₂ CH (CH ₂ ) i 0 3-ΝΟ ₂ -φ- -CHjCHj -CH3CH (CH3) j O 2-ΗΟ-φ- -ch, ch. -CH3CH (CH3) j 0 2-Naphthyl -CHjCHj -CH ₂ CH (CH ₂ ) ₂ 0 2,4-di-CM- -CH _; CHj -CH3CH (CH3) j 0 4-Br - -CH, CH, -CH, CH (CH3) i 0 3-Cl - -CH, CH, -CH ₂ CH (CH ₂ ) i 0 3Τ-Φ- -ch, ch. -CH3CH (CH3) j 0 benzothiazol-4-yl -CH, CH, -CH3CH (CH3) j O 2-CH, - <£ - -CH, CH, -CH ₂ CH (CH ₂ ) i 0 2-F-tt- -CHjCHj -CH _; CH (CH3) i 0 4-F-. -CH, CH, -CH, CH (CH3) i 0 3-Βγ-Φ- -CHjCHj -CH, CH (CH3) i 0 3-CF ₃ -4 -CHjCHj -CH ₃ CH (CH ₃ ) j 0 thien-2-yl -CHjCHj -CH ₂ CH (CH ₂ ) i 0 Φ- -CHjCHj -CH3CH (CH3) j O Φ- -CH (CH3) ₂ -CH ₃ CH (CH 2) CH ₃ CH, 0 Φ- -CHjCHjSCHj -CH3CH (CH3) j 0 Φ- -CH ₃ CH (CH ₃ ) j -CH3CH (CH3) j 0 3-α-φ- -CHj -CH, CH = C (CH 2) i 0 3-p - </> - -CHj -CH 3 -cyclopropyl 0 3-C1-4 »- -CHj -CH2-2-thienyl O 3-0Ι-φ- -CH, .CH, -CH 3 Cl 2 | ^ CH, CH, O 3-α-φ- -CH, -CH, -3-thienyl O

R * 1 ^R2 R ³ X 3-Cl-t / - -CH ₃ - (2-CH ₃ -cyclopentyl) O 3-Ο-φ- -ch ₃ -CH ₂ C = CH ₂ 1 ch ₃ O 3-Ο-φ- -CH, -cyclohex-2-enyl 0 2- [enyl- benzoxazol-5-yl -CH ₃ -CH ₂ CH (CH ₃ ) ₂ 0 3-CH ₃ S-é- -ch ₃ -CH, CH (CH _{3) 2} 0 furan-2-yl -ch ₃ -CH _; CH _{(CH3) 2} 0 benzophthiran-2-yl -CH, -CH ₂ CH (CH ₃ ) ₂ 0 benzothien-3-yl -CH ₃ -CH ₂ CH (CH ₃ ) i 0 2-chloro- thien-5-yl -CH ₃ -CH ₂ CH (CH ₃ ) ₂ O 3-methyl- isoxazol-5-yl -ch ₃ -CH ₂ CH (CH 3) ₂ O 2-4-S-thien-5-yl -ch ₃ -CH ₂ CH (CH ₃ ) ₂ O 6-CH3O- benzothiophen-2-yl -ch ₃ -CH ₂ CH (CH ₃ ) ₂ 0 3-phenyl-1,2,4-thiadiazol-5-yl -ch ₃ -CH ₃ CH (CH ₃ ) ₂ 0 2- (E-oxazol-4-yl) -CH, -CH ₂ CH (CH ₃ ) ₂ 0 3-CH3- © - -CH, -CH ₂ CH (CH ₃ ) ₂ 0 2,5-di-F- <£ - -ch ₃ -CH ₂ CH (CH 3) ₂ 0 3,5-di-F - -CHj -CH ₂ CH (CH ₃ ) ₂ 0 thien-3-yl -CH, -CH ₂ CH (CH 3) ₂ 0 4-CH ₃ - -ch ₃ -CH ₂ CH (CH 3) ₂ 0 Φ- -ch ₃ -CHCOOCHj 1 CH _{(CH3) 2} 0 3-NO ₂ - -ch ₃ -CH ₂ CH (CH ₃ ) ₂ 0

• *

R ' R ² R ³ X 3,5-di-F-d &gt;. -CH, -CH, CH, 0 3-ΝΟ _; -ψ- -CHiCHjSCH, -ch ₂ ch. 0 3-Cl-d -CH, -CH ₂ CH (CH ₂ ) ₂ 0 3-C1-0- -CH, -CH ₂ CH ₂ N (CH ₂ ) ₂ 0 3,5-di-Cl-O- -CHjCHjCHjCH, -CH, 0 3,5-di-Cl-O- -CH, -CH ₂ CH (CH ₂ ) ₂ 0 cyclohexyl -CH, -CH, CH (CH ₂ ) ₂ 0 cyclopentyi -CH, -CH ₂ CH (CH ₂ ) ₂ 0 cyclohex-1-enyl -CH, -CH ₂ CH (CH ₂ ) ₂ 0 3-Cl- - -CH, -CH ₂ CH = C (CH ₂ ) ₂ with 3,5-di-F-d -Φ -CH, 0 3,5-diF-d. - </> -CH ₂ CH (CH ₂ ) ₂ 0 cyclopentyi -Φ -CH, 0 cyclopentyi -CH, -CH, 0 cyclopropyl -Φ -CH, 0 cyclopropyl -CH, -CH, 0 3-ΝΟ ₂ -φ -CH3CH, SCH, -CH ₂ CH (CH ₂ ) ₂ 0

Other preferred compounds of formula I above include a compound wherein R ¹ is phenyl, R ² is fluoro, and R ³ is methyl.

Detailed description of the invention

As mentioned above, the present invention relates to compounds that inhibit β-amyloid peptide release

9 and / or its synthesis and accordingly can be used in the treatment of Alzheimer's disease. Before the present invention is described in 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 it may be, the β-amyloid peptide is a fragment of the larger transmembrane glycoprotein of approximately 39-43 amino acids, referred to as the β-amyloid precursor protein (APP). Its 43-amino acid sequence is as follows:

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 (SEC > ID NO: 1)

or a sequence that is substantially homologous to the sequence.

·· ·· * ·

9

9

99,999,999

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

Alkylene refers to divalent alkylene groups which preferably contain from 1 to 8 carbon atoms, and more preferably 1 to 6 carbon atoms. Examples include methylene (-CH ₂ -), ethylene (-CH ₂ CH ₂ ), propylene isomers (for example, -CH ₂ CH ₂ CH ₂ - and -CH (CH 3) CH ₂ -) and alike.

Alkoxy refers to an alkyl-O- group wherein alkyl has the same meaning as previously defined. 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. alike.

Alkylalkoxy refers to the group -alkylene-O-alkyl, wherein alkylene and alkyl are as defined above. Such groups include, by way of example, methylenethoxy (-CH ₂ OCH 3), ethylenethoxy (-CH ₂ CH ₂ OCH 3), npropylene-iso-propoxy (-CH ₂ CH ₂ CH ₂ OCH (CH ₃ ) ₂ ), methylene-tert-butoxy (-CH ₂ -OC (CH ₃ ) 3) and the like.

Alkylthioalkoxy refers to the group -alkylene-Salkyl, in which alkylene and alkyl have the meaning which was

9 9 9 9 9

9,999,999,999

9

999 99 99 99 defined above. Such groups include, by way of example, methylene-thiomethoxy (-CH ₂ SCH ₃ ), ethylene-thiomethoxy (-CH ₂ CH ₂ SCH ₃ ), n-propylene-iso-thiopropoxy (CH ₂ CH ₂ CH ₂ SCH (CH 3) 2), methylene-tert-thiobutoxy (- CH ₂ S (CH ₃ ) ₃ ) 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 preferably have 1 and more 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 (C = CH), propargyl (-CH ₂ C = CH) and the like.

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

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

Alkcycloalkyl refers to the group -alkylene-cycloalkyl, wherein alkylene and cycloalkyl are as defined above.

//

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

Acyloxy refers to alkyl-C (O) O-, aryl-C (O) O groups. heteroaryl-C (O) O- and heterocycle-C (O) O-, wherein the alkyl, aryl, heteroaryl and heterocyclic systems are as defined above.

Aryl refers to an unsaturated aromatic carbocyclic group containing from 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 an aryl substituent, such aryl groups may be optionally substituted with 1 to 3 substituents selected from the group consisting of hydroxy, acyl, acyloxy, alkyl, alkoxy, alkenyl, alkynyl, amino, aminoacyl, aryl, aryloxy, carboxyl, carboxylalkyl , acylamino, cyano, halogen, nitro, heteroaryl, trihalomethyl, thioalkoxy and the like. Preferred substituents include alkyl, alkoxy, halogen, cyano, nitro, trihalomethyl and thioalkoxy.

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

Carboxylalkyl refers to the group -C (O) O-alkyl, wherein the ones are: Alkyl has the same meaning as previously defined.

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 from 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 8 carbon atoms having a single cyclic ring and at least one internal unsaturation site, which may optionally be substituted with from 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 group containing 2 to 8 carbon atoms and 1 to 4 heteroatoms selected from oxygen, nitrogen and sulfur within the ring.

• · · 9

999 999

9

99

Unless otherwise limited by the definition of a heteroaryl 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, thioalkoxy, thioaryloxy. 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 and furyl.

A heterocycle or heterocyclic system refers to a monovalent (i.e., one point of attachment) saturated or unsaturated group having a single ring or multiple fused rings of from 1 to 8 carbon atoms and from 1 to 4 heteroatoms selected from the group consisting of nitrogen, a sulfur atom or an oxygen atom, within the ring.

Unless otherwise limited by the definition of a heterocyclic substituent, such heterocyclic groups may be optionally substituted with 1 to 3 substituents selected from the group consisting of alkyl, alkoxy, aryl, aryloxy, halogen, nitro. heteroaryl, thioalkoxy, thioaryloxy and the like. Such heterocyclic groups may have a single ring (e.g. piperidinyl or tetrahydrofuryl) or multiple fused rings (e.g. indolinyl, dihydrobenzofuran or quinuclidinyl). Preferred heterocycles include piperidinyl, pyrrolidinyl and tetrahydrofuryl.

• ftft ftft ftft ftft ftft ftft ftft ftft ftft ftft ftft

Examples of heterocycles and heteroaryls include, but are not limited to, furan, thiophene, thiazole, oxazole, benzothiazole, benzofuran, benzothiophene, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolizine, isoquinoline, quinoline, phthaline, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, phenanthroline, isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazolidine, imidazolidine, imidazolidine, imidazolidine, imidazolidine, imidazine alike.

Thiol refers to the -SH group.

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

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

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

The term pharmaceutically acceptable salt refers to a pharmaceutically acceptable salt of a compound of formula (I), which is derived from a number of organic and inorganic ions well known in the art and including, but not limited to: 0 0 0 0 0 0

0 00 000 000 000 0 0 0 0 0 0

By way of example only, sodium, potassium, calcium, magnesium ions, ammonium, tetraalkylammonium and the like; and when the molecule contains a basic functional group, also 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) above may be readily prepared by several divergent synthetic routes, the particular route being chosen due to the difficulty of compound preparation, the commercial availability of starting materials, and the like.

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

• · · · ·

wherein R ¹ , R ² , R ³ , X, X 'has been defined above.

and X have the same meaning as

Reaction (1) involves only the coupling of the appropriate acetic acid derivative [11] with the primary amine ester of the 2-amino acid ester under conditions that result in the formation of the N-acetyl derivative (2). can also be used to prepare the 2-amino acid N-acetyl esters of the present invention. For example, well-known coupling reagents such as carbodiimides or BOP (benzotriazol-1-yloxy-tris (dimethylamino) phosphonium hexafluorophosphate) can be used with the concomitant use of well known additives such as N-hydroxysuccinimide, 1-hydroxybenzotriazole and the like can be used to facilitate coupling . Reaction

..

is conveniently carried out in an inert aprotic diluent such as dimethylformamide, dichloromethane, chloroform, acetonitrile, tetrahydrofuran and the like. Alternatively, the hydrogen halide compounds 1 can be used in reaction (1) and, if used, are typically used in the presence of a suitable acid scavenging base formed during the reaction. Suitable bases include, by way of example, triethylamine, diisopropylethylamine, N-methylmorpholine and the like.

Reaction (1) is preferably carried out at a temperature of about 0 ° C to about 60 ° C until completion of the reaction, which usually occurs after about 1 to about 24 hours. After completion of the reaction, the 3-amino acid N-acetyl ester is isolated by conventional means, including precipitation, chromatography, filtration, and the like.

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

Alternatively, the synthesis described above in reaction (1) can be performed on the amino acid (XR ³ = OH) and after the formation of Nacetyl as described above, the carboxylic acid is then esterified with either alcohol (HOR ³ ) or thioalcohol (HSR ³ ) under conventional conditions The 3-amino acid N-acetyl ester, which is a compound of formula (I). For example, esterification procedures to obtain R ³ groups containing ester groups can be accomplished using the method described by Loss. et al. ¹⁰

In yet another embodiment of the present invention, conventional transesterification techniques can be used to prepare a variety of different ester groups on 3-amino acid N-acetyl esters. Numerous methods for carrying out transesterification are known in the art, and each such technique merely replaces the -OR group on the N-acetylester 2 amino acid with a different -OR ³ / -SR ³ group derived from the corresponding alcohol (i.e. HOR ³ ) or thioalcohol (e.g. is HSR ³ ) and in some cases a catalyst such as titanium isopropoxide is used to facilitate the completion of the reaction. In one method, the HOR ³ alcohol or HSR ³ thioalcohol is first treated with sodium hydride in a suitable diluent such as toluene to form the corresponding sodium alkoxide or thioalkoxide, which is then used to transesterify the 3 amino acid N-acetyl ester. The efficiency of this process makes it particularly effective when using high-boiling and / or expensive alcohols.

In another method of transesterification, the amino acid N-acetyl ester to be transesterified is placed in a large excess of alcohol or thioalcohol to be transesterified. Then, a catalytic amount of sodium hydride is added and the reaction proceeds rapidly under normal conditions and thus the transesterified product requires the use of a large one to obtain the desired.

Transesterification provides a simple way to create

7/2 f ························ A wide variety of substituents R ³ on the compounds of formula (I) above. In all cases, the alcohols and thioalcohols used to carry out the transesterification are well known in the art, and a significant number are commercially available.

Other methods for preparing esters of the present invention include, by way of example, first hydrolyzing the ester to the free acid, followed by O-alkylation with a halo-R ³ group in the presence of a base such as potassium carbonate.

The compounds described above can also be prepared using polymeric supported carbodiimide peptide coupling reagents. For example, EDC in polymer-supported form has been described (Tetrahedron Letters, 34 (48), 7685 (1993)) ¹⁵ . 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 have prepared side chains carrying 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 under the reaction conditions, e.g.

in which the final polymer bound coupling reagents will be used.

Certain hydroxymethylated resins may be converted to chloromethylated resins useful for preparing a coupling reagent on a polymeric support. Examples of such hydroxylated resins include 4-hydroxymethyl-phenylacetamidomethyl resin (Pam resin) and 4-benzyloxybenzyl alcohol resin (Wang resin) supplied by Advanced Chemtech of Louisville, Kentucky, USA (see Advanced Chemtech 19931994 catalog, page 115). The hydroxymethyl groups of these resins can be converted to the desired chloromethyl groups by any of a variety of methods well known to those skilled in the art.

Preferred resins are styrene / divinylbenzene resins, of easy commercial availability. As indicated, the resins are already chloromethylated due to their already chloromethylated name and require no chemical modifications before use. These resin resins are commercially known as Merrifields 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 P is an inert polymer and LG = Cl, Br, I or OSO ₂ CH ₃

Such methods are described more fully in U.S. Pat. Provisional Patent Application 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 with THF. The mixture to be prepared, CHCl ₃ and the concentration of 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.

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" refers to a physically discrete unit suitable as a single dose for humans

Or other mammals, each unit containing a predetermined quantity 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 preprocessed composition is then subdivided into unit doses of the type described above and which are of the type described above.

9 9 9 9 9

9 9 9 9 9 9 999 999 · · · · ·

9999 999 999 99 99 99 contain, for example, from about 0.1 to about 500 mg of the active ingredient of the present invention.

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 can be used to form such an enteric layer or coating, such as materials comprising a variety of polymeric acids and mixtures of polymeric acids with materials such as shellac, ketyl alcohol and cellulose acetate.

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.

ί # Γ

0 0 00 00 ♦ o · 0 0 0 0 0 0. 00 0000 000

0 0 0 0 0 0000 000 000 00 00 • · 0

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 produce a local or systemic 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.

Amount

Ingredient (mg / capsule)

Active ingredient 30.0

Starch 305.0

Magnesium stearate 5,0

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

9 9 9 9 9 9 9 9 9 9 · 9 9 9 9 9 9 9 9 9

999 99 99 99

Example of preparation 2

Tablets were prepared using the ingredients listed below:

Amount

Ingredient (mg / tablet)

Active ingredient 25.0

Cellulose, microcrystalline 200.0

Colloidal silica 10.0

Stearic acid 5,0

The ingredients were mixed and compressed into tablets, 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.

φ φ · φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ · · · ·

Example of preparation 4

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

Amount

Ingredient (mg / tablet)

Active ingredient 30.0 mg Starch 45.0 mg Microcrystalline cellulose 35.0 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 Overall 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.

• · · · · · · · · · · · · · · · · · · ·

Φ · · · φ · · · · · · · · · ·

Example of preparation 5

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

Component Amount (mg / capsule) Active ingredient 40.0 mg Starch 109.0 mg Magnesium stearate 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 quantity. 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 Quantity

Active Ingredient 50.0 mg

Xanthan gum 4.0 mg

Sodium carboxymethylcellulose (11%)

Microcrystalline cellulose (89%) 50.0 mg

Sucrose 1.75 g

Sodium benzoate 10.0 mg

Flavoring and coloring agent q.v.

Purified water to 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.

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• ·

Example of preparation 8

Component Amount (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:

The φφφφ folder

Amount

Active ingredient Emulsifying wax Liquid paraffin Soft white paraffin to

1-10 g 20 g

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 compositions 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 usually involve placement of a drug delivery catheter into the

the recipient's ventricular system that bypasses the blood-brain barrier. One such implantable delivery system used to transport biological factors 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, generally involve preparing the composition so as 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.

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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 the serum, the compounds may be encapsulated, inserted into lumenal liposomes, prepared as a colloid, or treated by another conventional technique that achieves prolonging the 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.

patient,

Conveniently,

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, based on factors such as the degree or severity of Alzheimer's disease in the age, weight and general condition of the patient and the like, when used as a therapeutic agent, the compounds of the present invention are administered in dosages. which are in the range of about 1 to about 500 mg / kg / day.

r ······································ · · ··············

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 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 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.

DETAILED DESCRIPTION OF THE INVENTION

In the discussion above and 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.

bm = wide multiplet

BOC = tert-butoxycarbonyl

BOP = benzotriazol-1-yloxy-tris (dimethylamino) phosphonium hexafluorophosphate bd = broad doublet bs = broad singlet

CDI = 1,1-carbodiimidazole d = doublet dd = doublet doublet dq = quartet doublet dt = triplet doublet

DMF = dimethylformamide

DMAP = dimethylaminopyridine

DMSO = dimethylsulfoxide

EDC = 1- (3-dimethyaminopropyl) -ethylcarbodiimide, hydrochloride eq. = equivalent

EtOAc = ethyl acetate g = gram h = hour

Hunig's base = diisopropylethylamine kg = kilogram = liter m = multiplet

M = molar

M% = mole percent max = maximum meq = milliequival mg = milligram ml = milliliter mm = millimeter mmol = millimol

N = normal ng = nanogram nm = nanometer

OD = optical density

P-EPC = 1- (3- (1-pyrrolidinyl) propyl) -3-ethylcarbodiimide psi = pounds per square inch Φ = phenyl q = quintet quint = quintet rpm - rpm s = singlet t = triplet

TFA = trifluoroacetic acid

THF = tetrahydrofuran tle = thin-layer chromatography μΐ = microliter

UV = ultraviolet • 9

9 9

9 9 9

9 ♦ 9 9

9

9999 999

In addition, the term Aldrich indicates that the compound or reagent used in the following examples is supplied by Aldrich Chemical Company, Inc., 1001 West Saint Paul Avenue, Milwaukee, WI 53233 USA;

the term Fluka indicates that the compound or reagent is supplied by Fluka Chemical Corp., 980 South 2nd Street, Ronkonkoma NY 11779 USA;

the term Lancaster indicates that the compound or reagent is supplied by Lancaster Synthesis, Inc., P.O. Box 100 Windham, NH 03087 United States;

the term Sigma indicates that the compound or reagent is supplied by Sigma, P.O. Box 14508, St. Louis MO 63178 United States;

the term Chemservice refers to the compound or reagent being supplied by Chemservice Inc, Westchester, PA, USA;

the term Bachem indicates that the compound or reagent is supplied by Bachem Bioscience Inc., 3700 Horizon Drive, Renaissance at Gulph Mills, King of Prussia, PA 19406 USA;

the term Maybridge indicates that the compound or reagent is supplied by Maybridge Chemical Co. Trevillett, Tintagel, Cornwall, United Kingdom PL34 OHW United Kingdom; and the term TCI indicates that the compound or reagent is supplied by TCI America, 9211 North Harborgate St., Portland, Oregon, 97203, OR, USA;

the term alpha indicates that the compound or reagent is supplied by the Johnson Maithey Catalog Company, Inc.

• 9 9 9 • 9 9 9

999 999

9

Bond Street, Ward Hill, MA 01835-0747 and the term Nova Biochem indicates that the compound or reagent is supplied by NovaBiochem USA, 10933 North Torrey Pines Road, P.O. Box 12087, La Jolla CA 92039-2087.

In the examples below, all temperatures are in degrees Celsius (unless otherwise indicated) and the following general procedures were used to prepare the compounds.

General procedure

Coupling of R ¹ C (X ') (X) C (O) Cl with H ₂ NCH (R ² ) C (O) 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 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.

General procedure

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 three times, once with brine, and once with brine. The solution of diethyl ether, washed with 10% HCl 20% once with potassium carbonate, was dried over magnesium sulfate, filtered, and evaporated under reduced pressure to thereby remove the solvent. product. Other amino acid esters may also be used in this procedure.

General procedure

Esterification R ^X 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

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 the temperature reached 999 ° C.

I · ·· ♦ did not indicate 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. 3NO2 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 dissolved in 15 ml H ₂ O. After stirring for 4 h, dioxane was removed under reduced pressure and 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 dried over Na ₂ SO ₄ and the solvent was removed under reduced pressure after filtration. The residue has been recrystallized •

9999 ··

I 9 9 9 »

9

99 from EtOAc / isooctane to give 7.5 g (85 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. [.alpha.] @ 23 = -29.9 @ 589 nm.

General procedure

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 backwashed with saturated aqueous acid portions were citric (2x), saturated sodium (2x), brine (1x), aqueous bicarbonate dried over anhydrous magnesium sulfate or sodium sulfate and the solvent was removed under reduced pressure to give the crude product.

General procedure F

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

0000

00 00 0 0

0 0 0 0 0 0

0 0 000 000

00 0 0 0 0 • 0 0 0 «000 000 0 0

Then 100 eq. 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

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. 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 purification by chromatography and / or recrystallization was required before biological experiments.

···

9

9 99 9 9 9

9

99

9 9 9 • · · · ··· ··· • ·

99

General procedure H

Coupling R ¹ C (X ') (X) C (0) Cl 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

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.

Specifically, a suitable 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 mL.

• ♦ φ • φ · · · φ φ φ φ.

Mg of β-EPC (0.87 meq / g) and the reaction 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 CHCl 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 anhydrous Na ₂ SO ₄ . 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.

Οι ₂ Η ₂₃ ΝΟ ₄ gives: C = 58.79, H = 9.38 and N = 5.71.

φ φ

φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ

Analysis found: C = 58.73, Η = 9.55, and Ν = 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.

C7H15NO2. 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

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

99

• 9

99 the reaction mixture was stirred at ambient temperature for 4 hours. The reaction mixture was then concentrated under reduced pressure to give 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)

♦ «·· · · · · · φ

Ě8 ·· · · · φ · · ··· · ·

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 sufficiently pure 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 were evaporated.

The desired 3,5-dichlorobenzyl methanesulfonate was obtained 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, •)

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.

Example 1

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 2

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:

^* H-NMR (CDC1 _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 = 7.1Hz, 3H), 0.91 (d, J = 6.7Hz,

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. Cl _S H ₂₃ NO ₃ (MW = 277.37, Mass Spectroscopy (MH ⁺ 278))

Example 3

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.

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 6.08 (d, J = 5.9 Hz, 1H), 4.62 (quint.,

J = 7.3 Hz, 1H), 3.92 (m, 2H), 2.22 (m, 1H), 1.84-2.00 (m, 3H), 1.40 (d, J = 7) 2 Hz, 3H), 1.35 (m, 1H), 1.20 (m, 1H), 0.85-0.96 (m, 12H).

¹³ C-nmr (CDCl 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 4

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 CO ₃ and aqueous HCl.

NMR data was as follows:

--nmr (CDCl ₃ ): δ = 7.30 (d, J = 8.2Hz, 2H), 7.21 (d, J = 8.3 Hz, 2H), 6.18 (d, J = 5.5 Hz, 1H) 4.57 (quint., J = 7.2 Hz, IH), 3.88 ( m, 2H) 3.53 (s, 2H). 1.91 (m, 1H), 1.36 (d, J = 7.1 Hz, 3 H) , 0.90 (d, J = 6.8Hz, 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. C15H20NO3CI (MW - 297.78, mass spectroscopy (MH ⁺ 298))

Example 5

55?

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 (CDCl 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, 1 H) 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, 131 3, 132 , 6, 134.7, 169,2,

172.8.

C15H19NO3Cl2 (MW = 332.23, mass spectroscopy (MH ⁺ 332))

Example 6

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

Following the procedure of 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 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): δ = 0.90 (d, J = 6.7 Hz (6H), 1.35 (d, J = 7.2 Hz, 3H), 1.91 (m, lH) , 2.34 (s, 3H) 3.55 (s, 2H), 3.88 (m, 2H) 4.58 (m, IH), 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, 7 1.3, 129.2, 129.6, 131,3, 136.9, 170.6, 172.8.

Η6Η ₂₃ Νθ3 (MW = 277.37, mass spectroscopy (MH ⁺ 278))

Example 7

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:

@ ¹ H-nmr (CDCl3): .delta. = 8.40 (d, J = 2 8, 2H); 7.6 (m, lH): 7.16 (m, 2 H); 4.5 (quint., J = 7.2, 7.2, IH) ; 3.8 (m, 2H); 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 03 (MW = 264, Mass Spectroscopy MH ⁺ 265))

• ·

Example 8

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:

¹ H-nmr (CDCl ₃ ): δ = 0.83 (m, 6H), 1.25 (d, J = 7.1 Hz, 3H), 1.81 (m, 1H), 3.79 (m (2H), 4.M (2s, 2H), 4.57 (quint., J = 7.3 Hz, 1H), 5.99 (d, J = 7.1 Hz, IH), 7.44 ( 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 9

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 having a melting point of 128 ° C-129 ° C. . The reaction was monitored by chromatography. · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · thin layer 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 = 0.86 (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 10

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.

99 99 9 9 9 9 ♦

99,999,999

9999 «99 999

99

C17H25NO3 (MW = 291.39, mass spectroscopy (MH ⁺ 292)).

Example 11

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: ¹ H-nmr (CDCl 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, I = 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.

C ₁₈ H ₂₇ NO ₃ (MW = 305, 39, mass spectroscopy (MH ⁺ 306)).

Example 12

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

Following the procedure of General Procedure F and using 4-pyridyl acetic acid in the form of hydrochloride (Aldrich) and (D, L) -alanine in the form of isobutyl ester hydrochloride (from Example B above), the compound was prepared from

7 / • 00 · 0 00 0

0 0 · 000

0 000 000 000

0 0 0 0

00000 00 00 of the title in the form of a solid having a melting point of 64 ° C-66 ° C. The reaction was monitored by thin layer chromatography on silica gel (Rf = 0.43 10% methanol / dichloromethane) and purification was performed by silica gel chromatography.

NMR data was as follows:

¹ H-nmr (CDCl 3): δ = 8.51 (dd, J = 1) r6, 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) , · O, , 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 = 2664, mass spectroscopy (MH ⁺ 26))

Example 13

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:

δ -nmr (CDCl ₃ ): δ = 7.24-7.39 (m, 5H), 6.14 (d, 1H), 4.58 (t, J = 7.3 Hz, 1H), 3, 88 (m, 2H), 3.58 (s, 2H), 1.90 (m, 1H), 1.35 (d, J = 7.2 Hz, 3H), 0.89 (d, J = 6) (7 Hz, 6H). ¹³ C-nmr (CDCl 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.

• 9 9 9 99 ·· ··

9 ·· 99 99 9999 · 999 9999 99 99 9999 999 999 · 9 9 9 9 9

9999 999 999 99 99 99

C15H21NO3 (MW = 263.34, mass spectroscopy (MH ⁺ 264)).

Example 14

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:

¹ H-nmr (CDCl 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 15

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 (CDCl 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, φ · «φ • • · · · · · · φ φ φ φ φ φ φ φ φ φ · · · · · · · φφ φφ φφ

2Η), 1.60 (m, 1Η), 0.64 (d, 6Η), 0.74 (t, 3Η). C ₁₇ H ₂₅ NO ₄ (MW = 307.39, mass spectroscopy (MH ⁺ 309)).

Example 16

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

Following the procedure of General Procedure I above and using

Of 4-nitrophenylacetic 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 = 8.16 (d, 2H), 7.44 (d, 2H), 6.04 (bd, 1H), 4.55 (m, 1H), 3.86 (dd, 2H) 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 17

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, 1H) 2H), 3.66 (s, 2H), 1.86 (m, 2H), 1.66 (m, 1H), 0.89 (d, 6H), 0.79 (t, 3H).

Example 18

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

Following the procedure of General Procedure I above and using

Of 3-thiopheneacetic 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 = 7.37 (m, lip, 7.16 (m, 1H), 7.04 (m,

1H), 6.05 (m, 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 19

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

Following the procedure of General Procedure I above and using

Of 4-chlorophenylacetic 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 = 7.22 (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).

Example 20

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 21

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, IH), 7.01 (m, IH), 6, 93 (m, IH), 6.7 9 (m, IH), 6.46 (m, IH), 4.51 (m, IH), 3, 87 (m, 2H), 3.57 (s, 2H), 2.01 (m, 2H), 1.75 (m, IH), 0.89 (d,

βΗ), 0.85 (t, 3Η).

Example 22

Synthesis of 2 - [(2-naphthyl) acetamidobutyric acid as isobutyl ester

Following general procedure I above and using 2-naphthylacetic 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 = 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 23

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 is

9

999 999 described in the general procedure.

NMR data was as follows:

¹ H-NMR (CDC1 _3): δ = 7.49 (m, l) e 7.22 (m, 21p 5.98 (m, press, 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 24

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 following: ¹ H-nmr (CDCl ₃ ) δ = 7.43 (d, 2H), 7.19 (d, 2H) 5.85 (m, 1H), 4.51 (m, 1H), 3.51 (m, 2H), 3.47 (with, 2H), 1.84 (m, 2H), 1.61 (m, 1 H) 0.84 (d, 6H) 0.76 (t, 3H). C ₁₆ H ₂₂ NO ₃ Br (MW = 356.26, mass spectroscopy (MH ⁺ 358))

Example 25

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 as follows:

¹ H-nmr (CDCl ₃ ): δ = 7.25 (m, 3H), 7.12 (m, 1H), 5.80 (m, 1H), 4.52 (m, 1H), 3.86 (m, 1H) m, 2H), 3.50 (s, 2H), 1.87 (m, 2H), 1.67 (m, 1H) 0.88 (d, 6H), 0.77 (t, 3H).

C16H22NO3Cl (MW = 311.81, mass spectroscopy (MH ⁺ 313)).

Example 26

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

Following general procedure 1 above and using 3-fluorophenylacetic 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 = 7.31 (m, 1H), 7.01 (m, 3H) 5.95 (m, 1H), 4.54 (m, 1H), 3.84 (m, 1H) m, 2H), 3.54 (s, 2H), 1.88 (m, 2H), 1.65 (m, 1H) 0.87 (d, 6H), 0.81 (t, 3H).

C16H22NO3F (MW = 295.35, mass spectroscopy (MH ⁺ 296)).

Example 27

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

Following the 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), 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 = 7.82 (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) ).

Example 28

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

Following general procedure I above and using 2-methylphenylacetic 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 - 7.18 (m, 410, 5.79 (m, lip, 4.54 (m,

1H), 3.85 (m, 2H), 3.59 (s, 2H), 3.29 (s, 3H), 1.51 (m,

2H), 1.59 (m, 1H), 0.87 (d, 6H), 0.77 (t, 3H).

C17H25NO3 (MW = 291.39, mass spectroscopy (M ⁺ 291)).

Example 29

Synthesis of 2 - [(2-fluorophenyl) acetamido] butyric acid as 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:

^* H-NMR (CDC1 _3): δ = 7.25 (m, 1H), 7.09 (m, 3H), 6.03 (m, 1H), 4.54 (m, 1H), 3.87 ( m, 2H), 3.57 (s, 2H), 1.89 (m, 2H), 1.64 (m, 1H), 0.88 (d, 6H), 0.80 (t, 3H).

Example 30

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, 6H), . 0.78 (t, 3H). C16H22NO3F (MW = 295.35, mass) spectroscopy (MH ⁺ 296).

Example 31

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

Following general procedure I above and using 3-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:

¹ H-nmr (CDCl ₃ ): δ = 7.45 (m, 2H), 7.23 (m, 2H) 5.95 (m,

1H), 4.55 (m, 1H), 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 32

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

Following general procedure I above and using 3-trifluoromethylphenylacetic 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 (CDC1 _3): δ = 7.52 (m, 1H) 7.47 (m, 2H) 6.01 1H), 4.56 (m, 1H), 3.86 (m, 2H), 3.61 (with, 2H), 1, 84 2H), 1.62 (m, 1H), 0.87 (d, 6H), 0.80 (t, 3H). C ₁₇ H ₂₂ NO ₃ F ₃ (MW = 345, 36, mass spectroscopy) (MH ⁺ 345))

Example 33

·····································

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 cleaning was carried out filtration, how Yippee described in the general procedure. NMR data was following: ¹ H-nmr (CDCl ₃ ) δ = 6.89 (m, 3H), 6.07 (bd, 1 H), 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). C14H21NO3S (MW = = 283.39, mass spectroscopy (MH ⁺ 284)).

Example 34

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)).

«Ft ft ft ·» »ft ft ft ft ft ft ft ft ft

Example 35

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 stirred for 18 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:

¹ 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, 1H), 3.47 (d, J = 13.8 Hz, 1 H), 2.05 (m, 1 H), 0.87 (d, J = 6.8, Hz, 3H), 0.84 (d, J = 6.8 Hz, 3) ¹³ C-nmr (DMSO) -dg): 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 φ φ • · · φ · · · · · · ·

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

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 7.25-7.40 (m, 5H), 5.95 (d, 1H), 4.56

(m, IH), 3.M-4 .00 (m, 2H). 3 , 61 (s, 2H), 2.10 (m, 1H), 1.68 (m, 1H), 1.38 (m, 1H), 1.15 (m IH) 0.82-0.94 (m, 9H), 0.76 (d, 3H). ¹³ c- • NMR (CDC1 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.

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

Example 36

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 evaporated under reduced pressure to an oil, the residue was diluted in

9 9

9 9 9 9 9

9 9 9 9 9

9999 999 999

9

999 99 99 99

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).

C17H25NO3S (MW = 323.17, mass spectroscopy (M ⁺ 323) dioxane (5.0 mL) bicarbonate)

Example 37

Synthesis of N- (phenylacetyl) -L-leucine as L-Leucine isobutyl ester (Aldrich) (0.1149, 0.869 mmol) was taken up 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:

¹ 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).

0 · 0 0 0 0

0 0 0 0 0 0

0 000 000 000

0 0 0 ·

000 00 00 00

C ₈ H ₂ 7NO ₃

0

0 0

0000 0 (MW = 305.40, mass spectroscopy (M ⁺ 305)).

Example 38

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.

NMR data was as follows:

Λ-nmr (CDCl ₃ ): δ = 7.39-7.16 (m, 4H), 6.06 (d, 1H), 5.385.29 (m, 1H), 4.63 (d, J = 9) Hz, 2H), 3.56 (s, 2H), 1.79 (s, 3H), 1.7 (s, 3H), 1.39 (d, J = 9 Hz, 3H)

Example 39

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:

δ -nmr (CDCl ₃ ): δ = 7.2-7.1 (m, 4H), 6.09 (s, 1H), 4.6 (dg, 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), 0.603-0.503 (m, 2H),

0.300-0.203 (m, 2 H).

Example 40

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.

NMR data was as follows:

¹ H-nmr (CDCl ₃ ): δ = 7.37-6, 97 (m, 7H), 5.97 (q, J = 14Hz, 2H), 4.6 (dq, J = 9Hz, 1H) ), 3.76 (s, 2H), 1.38 (d, J = 9 Hz, 3H).

Example 41

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

Following the general procedure C above and 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 carried out by liquid chromatography using 3: 7 EtOAc: hexane as the eluent.

NMR data was as follows:

^* H-NMR (CDC1 _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, 2H).

Example 42

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:

¹ H-nmr (CDCl ₃ ): δ = 8.03 (bd, J = 9 Hz, 1H), 7.56-7.5 (m, 1H), 7.47 (bs, 1H), 7.4 -7.17 (m, 4H), 7.06 (d, J = 9Hz, 1H), 5.1 (s, 2H), 4.3 (dq, 1H), 1.3 (d, J = 9 Hz, 3H).

Example 43

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

Following the general procedure C above 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:

¹ H-nmr (CDCl ₃ ): δ = 7.39-7.16 (m, 4H), 6.3 (bd, 1H), 4.79-

9 9 9 9 9 9 * 9 9999 ··· «99 99 9 9 9 9 9 9 9 9 9 9 9 • 9 «« 9 • 9 99 9 9 9,999 99 « 9 99 9 99 4.7 (m, 1 H), 4.6-4.25 (m, J = 9Hz, 1 H), 3,577 (WITH, 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, 1 H), 0.979 (dd, J = 9 Hz (2H) C ₇ H ₂ C1 2 NO ₃ (MW MS = 323.52, mass spectroscopy (MH ⁺ 323) • Example 44 Synthesis of N- [ (3-chlorophenyl) acetyl] alanine in the form 2-

of 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:

¹ H-nmr (CDCl ₃ ): δ = 7.39-7.16 (m, 4H), 6.03 (bs, 1H), 4.77 (s, 2H), 4.7- 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 45

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 by liquid chromatography using 3: 7 EtOAc: hexane

as the eluting solvent.

NMR data was as follows:

δ -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 = 9 Hz, 3H)

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

Example 46

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 general procedure I above) gave the title compound.

NMR data was as follows:

¹ H-NMR (CDC1 _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) 0.87 (d, 6H).

C22H24N2O4 (MW = 380, mass spectroscopy (MH ⁺ 381)).

Example 47

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:

δ -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 48

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 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 = 7.36 (m, IH), 6.34 (m, IH), 6.21 (m,

1H), 4.56 (m, 1H), 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 49

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

Following the general procedure I above using benzofuran-2-ylacetic acid (Maybridge) and alanine as isobutyl ester (prepared according to the general procedure

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 = 7.51 (m, 1H), 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 50

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

Following General Procedure I above and using thianaphthen-3-ylacetic acid (Lancaster) 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:

¹ H-nmr (CDCl ₃ ): δ = 7.89 (m, 1H), 7.76 (m, 1H), 7.38 (m,

3H), 6.07 (m, 1H), 4.57 (m, 1H), 3.92 (m, 2H), 3.82 (s,

4H), 1 M (m, 1H), 1.32 (d, 3H) 0.85 (d, 6H).

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

Example 51

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

Following the procedure of General Procedure I above and using

Qoo

5-chloro-2-thienylacetic acid (CAS # 13669-19-1 and alanine as isobutyl ester (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:

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)).

Example 52

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:

δ -nrar (CDCl ₃ ): 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 53

Synthesis of N - [(2-phenylthiothienyl) acetyl] alanine in the form of iso

9 99 9

9

99 butyl 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:

δ -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)).

Example 54

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

Following the procedure of 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:

^λ H-nmr (CDCl ₃ ): δ = 7.59 (d, 1H), 7.33 (d, 1H), 7.16 (s,

1H), 7.03 (dd, 1H), 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)).

• · 0 0 0 0 «··· • · · · · ·

0 0 0 9 99 999 9 9 · ·

102 ^× H-nmr (CDCl ₃ ): δ = 7.59 (d, 1H), 7.33 (d, 1H), 7.16 (s,

1H), 7.03 (dd, 1H), 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 55

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:

^* 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)).

Example 56

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

Following the procedure of General Procedure I above and using (2-phenyloxazol-4-yl) acetic acid (CAS # 22086-89-1) and alanine as isobutyl ester (prepared according to the 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.

··

A «· · · ········· · ·

- 103 Example 57

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)).

Example 58

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

Following General Procedure I above and using 2,5-difluorophenylacetic 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:

δ -nmr (CDCl 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, • 9) · 9 9 9 9

9 9 9 9 9

ΦΦΦΦΦΦ Φ ΦΦΦΦΦΦ ΦΦΦΦΦΦ ΦΦΦΦΦΦ · · · · · · · · ·

- 104 6).

C15H19NO3F2 (MW = 299, mass spectroscopy (h1H + 300)).

Example 59

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).

C15H19NO3F2 (MW = 299, mass spectroscopy (MH ⁺ 300)).

Example 60

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:

·

- 105 9 9 99 ^ -nmr (CDCl ₃ ): δ = 7.33 (m, 1H), 7.14 (m, IH), 7.01 (m, IH), 6.5 (m, IH) 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) @ 589 nm.

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

Example 61

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:

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).

C16H23NO3 (MW = 277.35, mass spectroscopy (MH ⁺ 278)).

Example 62

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

Following the procedure of 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)

106 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 charcoal (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:

¹ H-nmr (CDCl ₃ ): δ = 7.35-7.20 (m, 5H), 6.22 (bd, 1H), 4.83 (d, 1H), 4.65 (ρ, 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).

¹³ C-nmr (CDCl 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 63

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 99 0 0 0 99 00 0 99 00

107 purifications were carried out by recrystallization from butyl chloride.

NMR data was as follows:

δ -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: [α] 23 49 (c 5, MeOH).

Example 64

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 on silica gel using EtOAc as the eluting solvent followed by recrystallization from 1-chlorobutane. NMR data was as follows:

¹ H-NMR (DMSO-d _6): δ = 1.30 (d, 3H); 3.52 (s, 2 H).

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

Example 65

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. Ftft. Ftft. Ftft. Ftftftft. Ftft. Ftftftft. Ftft. Ftft. Ftft. Ftft. Ftft. ·

108 by recrystallization from butyl chloride. NMR data was as follows:

^λ 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 (s, 2H), 2.47 (t, 2H), 2.12 (m, 2H), 2.08 (with,

3H), 1.27 (t, 3H).

Optical Rotation: [α] 23-30 (c 5, MeOH).

Example 66

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 67

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

Following the general procedure C above and using N- (3-chlorophenylacetyl) alanine (from Example D above and 2- (N, N-dimethyl amino) ethanol (Aldrich), the title compound can be prepared. · Φ • · · · · · · · · · · · · φ φ φ φ φ φ φ φ φ

109 thin layer chromatography on silica gel and purification was carried out by liquid chromatography using 0.1: 2: 0.79 NH ₄ OH: EtOH: CHCl ₃ as the eluent.

NMR data was as follows:

1 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 1 H, 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 = 9Hz, 3H).

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

Example 68

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

Following the procedure of General Procedure F above and using

3,5-dichlorophenylacetic acid (from Example C above) and L-norleucine methyl ester hydrochloride (Bachem) gave the title compound 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.

NMR data was as follows:

δ -nmr (CDCl 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 (m), 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.M , 52.90, 43.14, 32.57,

27.87, 22.81, 14.41.

99 • 9 9 9

999 999

9

99

110

Example 69

Synthesis of N - [(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:

¹ H-nmr (CDCl ₃ ): δ = 7.27 (d, J = 2 Hz, 1H), 7.19 (s, 2H),

6.22 (d, J = 6Hz, 1H), 4.59 (quint., J = 7Hz, 1H), 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)).

Example 70

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

Following the procedure of General Procedure B above using cyclohexylacetic acid (Aldrich) and L-alanine as isobutyl ester hydrochloride (from Example B above), the title compound was prepared as a solid,

In cyclohexylacetic acid (Aldrich) and L-alanine as isobutyl ester hydrochloride (from Example B above), the title compound was prepared as a solid, m.p. 92-93 ° 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 (CDC1 _3): δ = 0.93 (d, J = 6.7 Hz, 6H), 0.85 to 1.01 (m, 2H), 1 05- 1.35 (m, 3H), 1.40 (d, J = 7.1 Hz, 3H), 1.601.85 (m, 6H), 1.95 (m, 1H), 2.06 (d, J = t 7.0 Hz) 2H, 3.92 (m, 2H), 4.61 (m, 1H), 6.08 (bd, 1H).

¹³ C-nmr (CDCl ₃ ): δ = 18.7, 18.9, 26, 0, 26, 1, 27.6, 33, 0,

35.3, 44.6, 47.9, 71.4, 171.8, 173.3.

C ₁₅ H ₂₇ NO ₃ (MW = 269, 39, mass spectroscopy (MH ⁺ 270)).

Example 71

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

NMR data was as follows:

99 · 9 · · ·

9 9 9 9 9

9 9 9999 999 999 · e e e e e e e e e e

- 112 -

^x H-nmr (CDC1 _3): δ = 0, 87 (d, J = 6.8 Hz, 6H), 1, 01 -1 17 (m, 2H), 1. 4 (d, J = 7.2) Hz, 3H), 1.40-1.62 (m, 4H), lr 70-1.83 (m, 2H) , 1.89 (m, 1H) 2.15 (m, 3H); 3.86 (m, 2H), 4 55 m IH), 6 30 (d, J = 7.1) Hz, 1H. ¹³ C-nmr (CDCl 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.

C14H25NO3 (MW = 255.36, mass spectroscopy (MH ⁺ 256))

Example 72

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 having a melting point of 49 ° C-50 ° C. The reaction was monitored by tlc on silica gel _(Rf = 0.40 in 1: 3 EtOAc: hexane) and purification was by extraction with Et ₂ O followed by washes with aqueous K ₂ CO ₃ and aqueous HCl.

NMR data was as follows:

^* H-NMR (CDC1 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). 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.

tt tt tt tt

113

Elemental Analysis

Calcd (%): C, 67.38; H, 9.42; N, 5.24; Found (%): C, 67.34; H, 9.54; N, 5.16.

C15H25NO3 (MW = 267.37, mass spectroscopy (MH ⁺ 268)).

Example 73

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

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:

^* H-NMR (DMSO-d₆): δ = 5.2 to 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).

C16H20NO2CIS (MW = 325, 86, mass spectroscopy (M ⁺ 325)).

Example 74

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 purification was performed by silica gel chromatography using a 1: 2 mixture.

- 114 ethyl acetate / hexanes as the eluting solvent.

NMR data was as follows:

1 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 75

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:

¹ H-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 76

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

3,5-difluorophenylacetic acid (Aldrich) was EDC coupled 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. Catalytic amount added • ·

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 1 mL of 1N HCl and saturated aqueous NaHCO 3 was added after stirring for a few minutes. 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, 5H) 6.8 -6.7 (m, 3H) ), 6.60 (d IH, 7 Hz), 5.55 (d IH 7.1 Hz), 3.9 (m 2H), 3.M (with 2H), 1.85 (m 1 H (7 Hz), 0.8 (q 6H 7 Hz) ¹³ C-nmr (CDCl 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 77

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

116 (m, 2H) ¹³ C-NMR (CDC1 _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 78

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:

δ -nmr (CDCl 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 79

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:

δ -nmr (CDCl ₃ ): δ = 7.35 (m, 5H) 6.97 (M, J = 7.2 Hz, 1H) 5.59 (d, J = 7.8 Hz, IH), 3 71 (s, 3H), 2.17 (m, 2H), 1.050.95 (m, 1H), 0.62 (m, 2H), 0.02 (m, 2H).

• «· · · · · ·» • · · · · · · · «« · · · · - 117 - β » ·· · • • · 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 Example 80 Synthesis N- (cyclopropylacetyl) -L-alanine ve form

methyl ester

Following the procedure of General Procedure D above and using cyclopropylacetic 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 (s,

3H), 2.10 (m, 2H), 1.34 (d, 3H), 0.95 (m, 1H), 0.58 (m, 2H) 0.15 (m, 2H) ¹³ C-nmr (CDC1 _3): δ = 173.7, 172.3, 52.3, 47.7, 41.0, 18.2, 6.7, 4.27, 4.22

Example 81

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:

δ -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, 2H); 2.15 (m, 1 H), 2.02 (s, 3H), 1.90 (m, 2H), 0.91 (d, 6H).

·

- 118 * 99 · 9 · A · A · A · A · A

C17H24N2O5S (MW = 368.4, mass spectroscopy (MH ⁺ , 368)).

Example 82

Cell search for the detection of inhibitors of βamyloid production

A number of the compounds of formula (I) above have been tested for their ability to inhibit β-amyloid formation in cells containing a Swedish mutation. This screening analysis utilized cells (K293 - human kidney cell lines) that were stably transfected with the amyloid precursor protein 751 gene (APP751) containing the double mutation Lys65iMet ₆₅₂ to Asn ₆ 5iLeu652 (at APP751 numbering) as described in the international patent publication. No. 94/10569 ⁸ and in Lemon et al. ¹¹ . This mutation is commonly referred to as the Swedish mutation, and the cells, designated "293,751 SWE, were plated in Corning 96-well plates at 1.5-2.5 x 10 ⁴ cells per well in Dulbecco's minimum essential medium with the addition of 10% fetal 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 overnight incubation at 37 ° C in an incubator equilibrated with 10% carbon dioxide, the media was removed and replaced with 200 ml of compound of formula (I) containing the medium per well for two hours pretreatment and cells were incubated as above.

· «« 9 9 9 «9« «« «« «« «

119

Drug stock solutions were prepared in 100% dimethylsulfoxide such that at the final drug concentration used in the processing, the dimethylsulfoxide concentration did not exceed 0.5% and was in fact usually 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 ml of treated medium or a suitable dilution transferred to ELISA plates coated with antibody 266 ¹³ against amino acids 13-28 of β-amyloid peptide, which is disclosed in International Patent Application Nos. 94/10569 ⁸ and stored at 4 ° C overnight. An ELISA assay employing labeled antibody 6C6 ¹³ 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., ¹² . 25 ml of a stock solution (5 mg / ml) of 3, (4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) was added to the cells remaining in the plate culture to a final concentration of 1 mg / ml. ml. 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).

• ·

9 98 9 ·

• * · *

Sodium dodecyl sulfate in 50% dimethylformamide, pH 4.7). Complete extraction was achieved by shaking overnight at ambient temperature. Unlike was measured and OD 62nni _{5 6} 50 nM in a microplate reader, Molecular Devices UV _max as an indicator of 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 83

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

• · · ·

0 0 0 0 0 0 0 0

0 0 ·· 0 · 0 · 00

9 · · · 000 000 · * ··

The 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 various vehicles such as corn oil (Safeway, South San Francisco, CA); 10% EtOH in corn oil (Safeway); 2-hydroxypropyl-β-cyclodextrin (Research Biochemicals 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 by CO ₂ anesthesia and blood was collected by cardiac puncture using a 1 cc 25G 5/8 tuberculin syringe / needle 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.

- 122 · · 122 122 122 122 122 122 122 122 122 122 122 122 122 122 122 122 122 122 122 122 122

Brain homogenates were diluted 1:10 with ice-casein buffer [0.25% casein, phosphate buffered saline (PBS), 0.05% sodium azide, 20 mg / ml aprotinin, 5 mM EDTA, pH 8.0, 10 mg / ml 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 1-40 or 1-42) were prepared so 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. The capture antibody, 266 ¹³ , is specific for amino acids 13 -28 β-amyloid. 3D6 ^14, 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.

Configuring the β-level sandwich ELISA

Amyloid 123 (amino acids 1-42) uses monoclonal antibody 21F12 ¹⁴ (which recognizes amino acids 33-42 of 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 plated at 10 mg / 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 mg / 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.

- 124 • * e · φ

• • e · ·

0 00 00 9 9 9 · 9 ·

9 9 99 9 9

9 · ·

9 9 · · ·

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 the total b-amyloid assay (266 capture antibody / 3D6 indicator antibody) described above for the brain assay except that a sample diluent was used instead of 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.

- 125

LIST OF SEQUENCES (1) GENERAL INFORMATION:

(i) APPLICANTS:

ATHENA NEUROSCIENCES, INC.

ELI LILLY & COMPANY

JING WU

EUGENE D. THORSETT JEFFREY S. NISSEN THOMAS MABRY LEE H. LATIMER VARGHESE JOHN

LAWRENCE Y. FANG JAMES E. AUDIA (ii) NAME OF THE INVENTION:

N- (ARYL / HETEROARYLACETYL)

AMINO ACID ESTERS, PHARMACEUTICAL COMPOSITIONS THAT CONTAIN THEM AND METHODS OF INHIBITION AND / OR ITS RELEASE RELEASE OF β-AMYLOID PEPTIDE

SYNTHESIS USING SUCH COMPOUNDS (iii) NUMBER OF SEQUENCES: 1 (iv) CORRESPONDENCE ADDRESS:

(A) ADDRESS: Burns, Doane, Swecker & Mathis, LLP (B) STREET: P.O. Box 1404 (C) CITY: Alexandria

- 126

• (D) COUNTRY: Virginia (E) COUNTRY: U.S.A.

(F) ZIP Code: 22313-1404 (v) COMPUTER FORMAT:

(A) MEDIA TYPE: Flexible Disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS / MS-DOS (D) SOFTWARE: PatentIn Release # 1.0,

Version # 1. 30 (vi) CURRENT APPLICATION DATA:

(A) APPLICATION NUMBER: Not Assigned (B) DATE OF ADMINISTRATION: Not Assigned (C) CLASSIFICATION:

(vii) PREVIOUS APPLICATION DATA:

(A) U.S. Pat. PATENT APPLICATION NO. 08 / 754,895 (B) filing date: 22 November 1996 (viii) PATENT REPRESENTATIVE INFORMATION:

(A) NAME: Swiss, Gerald F.

(B) REGISTRATION NUMBER: 30.113 (C) REFERENCE / DOCUMENT NUMBER: 002010-051 (ix) TELECOMMUNICATION INFORMATION:

(A) PHONE: 650-77377-854-7400 (B) PHONE: 650-77377-854-8275 (2) INFORMATION FOR SEQ ID NO: 1:

127 · · · t · «« ((((((((((((((((((( (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 43 amino acids (B) TYPE: peptide (D) TOPOLOGY: linear (ii) MOLECULA TYPE: peptide

(xi; 1 DESCRIPTION SEQ ID NO NO 1: Asp 1 Ala Glu Phe Arg His Asp 5 Ser Gly Tyr Glu His His 10 Gin 15 Dec Lys Leu Val Phe 20 May Glu Asp Gly Ser Asn Lys 25 Gly 30 Ala Ile Ile Gly Leu Met 35 Val Gly Val Val Ile Ala 40

Thr

Represented by:

dr. Otakar Švorčík

Claims (2)

00 * · yl, thien-3-yl, benzothiazol-4-yl, 2-phenylbenzoxazol-5-yl, furan-2-yl, benzofuran-2-yl, benzothiophen-3-yl, 2-chlorothien-5-yl, 3-methylisoxazol-5-yl, 2- (phenylthio) thien-5-yl, 6-methoxythiophen-2-yl, 3-phenyl-1,2,4-thiooxadiazol-5-yl and 2-phenyloxazol-4-yl. 63. The pharmaceutical composition according to claim 47, 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, phenyl and alkylthioalkoxy containing 1 to 4 carbon atoms. 64. The pharmaceutical composition according to claim 63, characterized in that R ² is selected from the group consisting of methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, -CH2CH2SCH3 and phenyl. 65. The pharmaceutical composition of claim 47 wherein X ' and X are both hydrogen and X is oxygen. 66. The pharmaceutical composition according to claim 65, characterized in that R ³ is selected from the group consisting of methyl, ethyl, isopropyl, n-propyl, n-butyl, iso-butyl, cyclopentyl, allyl, isobut-2-enyl, 3 methylpentyl, -CH ₂ -cyclopropyl, -CH ₂ -cyclohexyl, CH _2- (3-tetrahydrofuranyl), _{-CH2-thien-2-yl,} -CH ₂ - (1- methyl) cyclopropyl, _-CH2-thien-3 -yl, -CH ₂ -C (O) O-tert-butyl, -CH ₂ C (CH ₃ ) ₃ , -CH ₂ CH (CH ₂ CH ₃ ) 2, -2-methylcyclopentyl, -cyclohex-2- enyl, CH [CH (CH ₃ ) ₂ ] COOCH ₃ , -CH ₂ CH ₂ N (CH ₃ ) ₂ , -CH ₂ C (CH ₃ ) = CH ₂ and -CH ₂ CH = C (CH ₃ ) ₂ . 67. The pharmaceutical composition of claim 47 wherein X1 and X are both hydrogen and X is sulfur. 68. A pharmaceutical composition according to claim 67, characterized in that R ³ is selected from the group consisting of iso-but-2-enyl, and isobutyl. - 152 69. The pharmaceutical composition of claim 47 wherein the compound of formula I is selected from the group consisting of: N- (phenylacetyl) alanine, iso-butyl ester N- (3-phenylpropionyl) alanine, iso-butyl ester N- (3-methylpentanoyl) alanine, iso-butyl ester N - [(4-chlorophenyl) acetyl] alanine, isobutyl ester N - [(3,4-dichlorophenyl) acetyl] alanine, iso-butyl ester N - [(3-pyridyl) acetyl] alanine, isobutyl ester N - [(1-Riaphthyl) acetyl] alanine, iso-butyl ester N - [(2-naphthyl) acetyl] alanine, iso-butyl ester N- (4-phenylbutanoyl) alanine, iso-butyl ester N- (5-phenylpentanoyl) alanine, iso-butyl ester N - [(4-pyridyl) acetyl] alanine, isobutyl ester 2 - [(3,4-dichlorophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(3-methoxyphenyl) acetamido] butyric acid, iso-butyl ester 2 - [(4-nitrophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(3,4-methylenedioxyphenyl) acetamido] butyric acid, iso-butyl ester 2 - [(thien-3-yl) acetamido] butyric acid, iso-butyl ester 2 - [(4-chlorophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(3-nitrophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(2-hydroxyphenyl) acetamido] butyric acid, iso-butyl ester 2 - [(2-naphthyl) acetamido] butyric acid, iso-butyl ester 2 - [(2,4-dichlorophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(4-bromophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(3-chlorophenyl) acetamido] butyric acid, 2 - [(3-fluorophenyl) acetamido] butyrate, iso-butyl ester 2- [(benzothiazol-4-yl) acetamido] butyric acid, iso-butyl ester - 153 · 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 - [(2-methylphenyl) acetamido] butyric acid, iso-butyl ester 2 - [(2-fluorophenyl) acetamido] butyric acid, 2 - [(4-fluorophenyl) acetamido] butyrate, iso-butyl ester 2- [(3-bromophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(3-trifluoromethylphenyl) acetamido] butyric acid, iso-butyl ester 2 - [(2-thienyl) acetamido] butyric acid, iso-butyl ester 2- (phenylacetamido) butyric acid, iso-butyl ester N- (phenylacetyl) valine, 2-methylbutyl ester N- (phenylacetyl) methionine, iso-butyl ester N- (phenylacetyl) leucine, iso-butyl ester N - [(3-chlorophenyl) acetyl] alanine, 3-methylbut-2-enyl ester N - [(3-chlorophenyl) acetyl] alanine, cyclopropylmethyl ester N - [(3-chlorophenyl) acetyl] alanine, 2-thienylmethyl ester N - [(3-chlorophenyl) acetyl] alanine, (1-methylcyclopropyl) methyl ester N - [(3-chlorophenyl) acetyl] alanine, 3-thienylmethyl ester N - [(3-chlorophenyl) acetyl] alanine, 2-methylcyclopentylester N - [(3-chlorophenyl) acetyl] alanine, 2-methylprop-2-enyl ester N - [(3-chlorophenyl) acetyl] alanine, cyclohex-2-enyl ester N - [(2-phenylbenzoxazol-5-yl) acetyl] alanine, iso-butyl ester N - [(3-methylthiophenyl) acetyl] alanine, isobutyl ester N-4 - [(2-furyl) acetyl] alanine, iso-butyl ester N - [(benzofuran-2-yl) acetyl] alanine, iso-butyl ester N - [(benzothiophen-3-yl) acetyl] alanine, iso-butyl ester N - [(2-chloro-5-thienyl) acetyl] alanine, iso-butyl ester N - [(3-methyl-isoxazol-5-yl) acetyl] alanine, iso-butyl ester N - [(2-phenylthiothienyl) acetyl] alanine, iso-butyl ester N - [(6-methoxybenzothiophen-2-yl) acetyl] alanine, isobutyl ester N - [(3-phenyl-1,2,4-thiadiazol-5-yl) acetyl] alanine, iso-butyl ester N - [(2-Phenyloxazol-4-yl) acetyl] alanine, iso-butyl ester N - [(3-methylphenyl) acetyl] alanine, iso-butyl ester N - [(2,5-difluorophenyl) acetyl] alanine, iso-butyl ester N - [(3,5-diflurophenyl) acetyl] alanine, iso-butyl ester ··· ΦΦΦ ΦΦΦ ΦΦ · ΦΦ φφ φ - 154 Ν - [(3-thienyl) acetyl] alanine, iso-butyl ester Ν - [(4-methylphenyl) acetyl] alanine, iso-butyl ester N- (phenylacetyl) alanine, (1-methoxycarbonyl) isobutyl ester N - [(3-nitrophenyl) acetyl] alanine, iso-butyl ester N - [(3,5-difluorophenyl) acetyl] alanine, ethyl ester N - [(3-nitrophenyl) acetyl] methionine, ethyl ester N - [(3-chlorophenyl) acetyl] alanine, isobutyl ester N - [(3-chlorophenyl) acetyl] alanine, 2- (N, N-dimethylamino) ethyl ester 000 0 · · · 0 000 0 0000 0 0 0 · ·· · ·· JUDr. Otakar Švorčík - 128 ·· ·· .. · .. .. - · · ··· ·. ·. ··· · ··· «·», lawyer * · · · · ······· · · · • · · · · · · · · · · · · Halkova 120 00, Prague 2 ················· PAT E Ν T OVÉ Claims: 1. A compound of general formula formulas III: X in X ” 0 \ / H II . HI Ύ F R2 XR ³ / in which R ¹ is selected from the group consisting of a) alkyl, alkenyl, alkcycloalkyl, phenyl- (R) _m , -, naphthyl- (R) _m , wherein R is an alkylene group having 1 to 8 carbon atoms and _m is an integer equal to 0 or 1, cycloalkyl, cycloalkenyl 3-pyridyl, 4-pyridyl and heteroaryl, other than 3- and 4-pyridyl, having from 3 to 10 atoms and from 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen, wherein the heteroaryl group is optionally substituted with 1 to 4 3 substituents selected from the group consisting of alkyl, alkoxy, aryl, aryloxy, halogen, nitro, thioalkoxy and thioaryloxy, provided that if such heteroaryl contains at least one nitrogen heteroatom, it also contains at least one oxygen and / or sulfur heteroatom ; (b) a substituted phenyl group of formula (II): - 129 in which R is alkylene of 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, cyano, cycloalkyl, halogen, heteroaryl, 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 or 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- (R) _m -, wherein R is a C 1 -C 8 alkylene group and _m is an integer equal to 0 or 1, substituted at the 5, 6, 7 position and / or or with 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, phenyl, alkylalkoxy, alkylthioalkoxy; and R ³ is selected from the group consisting of - (CH ₂ ) _n CR ¹⁰ R ⁵ R ⁶ , wherein n is an integer equal to 0, 1 or 2, R ⁵ and R ⁶ are independently 130 is selected from the group consisting of hydrogen, alkyl, alkenyl, aryl, heteroaryl, heterocyclic, -NR ⁷ R ⁸ wherein R ⁷ and R ⁸ are each independently hydrogen or alkyl and -COOR ⁹ wherein R ⁹ represents wherein R ⁵ and R ⁶ may be joined to form a cycloalkyl group, a cycloalkenyl group, an aryl group, a heteroaryl group and a heterocyclic group, and if R ⁵ and R ⁶ are not joined to form an aryl or heteroaryl group, R ¹⁰ is selected from from the group consisting of hydrogen and alkyl with the proviso that if n is zero, then R ¹⁰ represents a hydrogen atom and if n is greater than zero and R ⁵ and R ⁵ are joined to form an aryl or heteroaryl group, then R ¹⁰ represents a bond within this group; X represents an oxygen atom or a sulfur atom; 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, and their pharmaceutically acceptable salts, with the additional conditions that: when R ¹ represents phenyl, R ² represents -CH (CH 3) CH ² CH 3, X represents an oxygen atom and X 1 and X represent a hydrogen atom, then R ³ does not represent -CH 2 CH 3 or -CH 2 CH (CH ₃ ) ₂ ; when R ¹ is phenyl, R ³ is -CH 2 CH (CH 3) 2, X is oxygen and X 1 and X are hydrogen, then R ^{2 is} not -CH (CH 3) ₂ when R ¹ is pyrid-3-yl, R ² is ethyl, X is oxygen and X 1 and X are hydrogen, then R ^{3 is} not -CH ₂ CH (CH ₃ ) 2 when R ¹ is indoxazin-3-yl, 2,4-dimethylthiazole-5- yl, 4-methyl-1,2,5-thíooxadizol-3-yl or 3,5-di (trifluoromethyl) phenyl, R ² is methyl, X is oxygen, and X 'and X are hydrogen, then R ³ does not represent -CH 2 CH (CH3) 2, and where ^R1 is -CH2-phenyl, ^R3 is -CH2CH3, X is oxygen, and X 'and X are hydrogen, then R ² does not represent -CH 2 CH (CH ₃₎ 2, • · · 131 and in addition with the further condition that the following known compound is excluded: N- (phenylacetyl) methionine ethyl ester. A compound according to claim 1, an unsubstituted phenyl group. in which R ¹ represents A compound according to claim 1, in an unsubstituted naphthyl group comprising 1-naphthyl and 2-naphthyl. which R ¹ represents selected from the group, A compound according to claim 1, wherein R ^{1 is a} substituted phenyl group of formula II. represents A compound according to claim 4, wherein the substituted phenyl group is defined as follows: (a) monosubstituted phenyls having a single substitution at the 2, 3 or 4 position, wherein each of the individual substituents is determined by the corresponding R ^a , R ^b and R ^c groups; 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, wherein each of the substituents is determined by the corresponding groups R ¹ , 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. and wherein again each of the substituents is determined 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, 3-32, 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 1, wherein R ¹ represents a phenyl- (R) _m - group, wherein R is alkylene containing from 1 to 8 carbon atoms and _m is 1. The compound of claim 7, wherein R ¹ is selected from benzyl, 3-phenyl-n-propyl and 4-phenyl-n-butyl. The compound of claim 1, wherein R ¹ is selected from the group consisting of alkyl, alkenyl, alkcycloalkyl, cycloalkyl and cycloalkenyl. 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. The compound of claim 9, wherein R ¹ is cycloalkenyl. The compound of claim 9, wherein the alkyl, cycloalkyl, alkcycloalkyl, alkenyl and cycloalkenyl groups R ¹ 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. The compound of claim 1, wherein R ¹ is selected from the group consisting of heteroaryl and substituted heteroaryl groups. ♦ · · • · Β Β Β Β · · · · · · · · · · · · · · · · · · · - 133 thien-3-yl, furan-2-yl, The compound of claim 15, wherein the heteroaryl and substituted heteroaryl groups R ¹ are selected from pyrid-3-yl, pyrid-4-yl, thien-2-yl, benzothiazol-4-yl, 2-phenylbenzoxazole- 5-yl, benzofuran-2-yl, benzothiophen-3-yl, 2-chlorothien-5-yl, 3-methylisoxazol-5-yl, 2- (phenylthio) thien-5-yl, 6-methoxythiophen-2-yl, 3-phenyl -1,2,4-thiooxadiazol-5-yl and 2-phenyloxazol-4-yl. A compound according to claim 1, 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, phenyl and alkylthioalkoxy of 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, phenyl, and -CH 2 CH 2 SCH 3. The compound of claim 1, wherein X 'and X are both hydrogen and X is oxygen. A compound according to claim 19, wherein R ³ is selected from the group consisting of methyl, ethyl, isopropyl, n-propyl, n-butyl, isobutyl, cyclopentyl, allyl, iso-but-2-enyl, 3-methylpentyl , -CH ₂ cyclopropyl, -CH ₂ -cyclohexyl, -CH ₂ - (3-tetrahydrofuranyl), -CH ₂ thien-2-yl, -CH ₂ - (1-methyl) cyclopropyl, -CH ₂ -thien-3- yl, -CH ₂ C (O) O -tert. -butyl, -CH ₂ -C (CH ₃ ) ₃ , -CH ₂ CH (CH ₂ CH ₃ ) 2, -2-methylcyclopentyl, -cyclohex-2-enyl, -CH [CH (CH ₃ ) ₂ ] COOCH ₃ , CH ₂ CH ₂ N (CH ₃ ) 2, -CH ₂ C (CH ₃ ) = CH _2, and -CH ₂ CH = C (CH ₃ ) 2. The compound of claim 1, wherein X 1 and X are both hydrogen and X is sulfur. The compound of claim 21, wherein R ³ is selected from iso-but-2-enyl and iso-butyl. • · · - 134 · φ φ φ 134 134 134 134 134 A compound according to claim 1, wherein the compound of formula I is selected from the group consisting of: N- (phenylacetyl) alanine, iso-butyl ester N- (3-phenylpropionyl) alanine, iso-butyl ester N- (3-methylpentanoyl) alanine, iso-butyl ester N - [(4-chlorophenyl) acetyl] alanine, isobutyl ester N - [(3,4-dichlorophenyl) acetyl] alanine, iso-butyl ester N - [(3-pyridyl) acetyl] alanine, iso-butyl ester N - [(1-naphthyl) acetyl] alanine, iso-butyl ester N - [(2-naphthyl) acetyl] alanine, iso-butyl ester N- (4-phenylbutanoyl) alanine, iso-butyl ester N- (5-phenylpentanoyl) alanine, iso-butyl ester N - [(4-pyridyl) acetyl] alanine, isobutyl ester 2 - [(3,4-dichlorophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(3-methoxyphenyl) acetamido] butyric acid, iso-butyl ester 2 - [(4-nitrophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(3,4-methylenedioxyphenyl) acetamido] butyric acid, iso-butyl ester 2 - [(thien-3-yl) acetamido] butyric acid, iso-butyl ester 2 - [(3-nitrophenyl) acetamido] butyric acid, 2 - [(3-nitrophenyl) acetamido] butyric acid, 2 - [(2-hydroxyphenyl) acetamido] butyric acid, iso-butyl ester 2 - [(2-naphthyl) acetamido] butyric acid, iso-butyl ester 2 - [(2,4-dichlorophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(4-bromophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(3-chlorophenyl) acetamido] butyric acid, 2 - [(3-fluorophenyl) acetamido] butyrate, iso-butyl ester 2- [(benzothiazol-4-yl) acetamido] butyric acid, 135 ··· 4 · · · · · · · · · · · · I · Isobutyl ester 2 - [(2-methylphenyl) acetamido] butyric acid, iso-butyl ester 2 - [(2-fluorophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(4-fluorophenyl) acetamido] butyric acid, 2 - [(3-bromophenyl) acetamido] butyrate, iso-butyl ester 2- [(3-trifluoromethylphenyl) acetamido] butyric acid, iso-butyl ester 2 - [(2-thienyl) acetamido] butyric acid, 2- (phenylacetamido) butyric acid iso-butyl ester, N- (phenylacetyl) valine iso-butyl ester, 2-methylbutyl ester N- (phenylacetyl) methionine, iso-butyl ester N- (phenylacetyl) leucine, iso-butyl ester N - [(3-chlorophenyl) acetyl] alanine, N - [(3-chlorophenyl) acetyl] alanine, N - [(3-chlorophenyl) acetyl] alanine, N - [(3-chlorophenyl) acetyl] alanine, 2-thienylmethyl ester N- [ (3-chlorophenyl) acetyl] alanine, (1-methylcyclopropyl) methyl ester N - [(3-chlorophenyl) acetyl] alanine, 3-thienylmethyl ester N - [(3-chlorophenyl) acetyl] alanine, 2-methylcyclopentyl ester N - [(3-chlorophenyl) acetyl] alanine, 2-methylprop-2-enyl ester N N - [(2-phenylbenzoxazol-5-yl) acetyl] alanine, N - [(3-methylthiophenyl) acetyl] alanine, isobutyl ester, - [(3-chlorophenyl) acetyl] alanine, cyclohex-2-enyl ester N-4 - [(2-furyl) acetyl] alanine, iso-butyl ester N - [(benzofuran-2-yl) acetyl] alanine, iso-butyl ester N - [(benzothiophen-3-yl) acetyl] alanine, iso-butyl ester N - [(2-chloro-5-thienyl) acetyl] alanine, iso-butyl ester N - [(3-methyl-isoxazol-5-yl) acetyl] alanine, isobutyl ester N - [(2-phenylthiothienyl) acetyl] alanine, isobutyl ester N - [(6-methoxybenzothiophen-2-yl) acetyl] alanine, isobutyl ester N - [(3-phenyl-1,2,4-thiadiazol-5-yl) acetyl] alanine, iso-butyl ester N - [(2-Phenyloxazol-4-yl) acetyl] alanine, iso-butyl ester 136 φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ Ν - [(3-methylphenyl) acetyl] alanine, iso-butyl ester Ν - [(2,5-difluorophenyl) acetyl] alanine, iso-butyl ester N - [(3,5-diflurophenyl) acetyl] alanine, isobutyl ester N - [(3-thienyl) acetyl] alanine, iso-butyl ester N - [(4-methylphenyl) acetyl] alanine, isobutyl ester N- (phenylacetyl) alanine, (1-methoxycarbonyl) isobutyl ester N - [(3-nitrophenyl) acetyl] alanine, iso-butyl ester N - [(3,5-difluorophenyl) acetyl] alanine, ethyl ester N - [(3-nitrophenyl) acetyl] methionine, ethyl ester N - [(3-chlorophenyl) acetyl] alanine, isobutyl ester N - [(3-chlorophenyl) acetyl] alanine, 2- (N, N-dimethylamino) ethyl ester 2 - [(3,5-dichlorophenyl) acetamido] hexanoic acid methyl ester N - [(3,5-dichlorophenyl) acetyl] alanine, iso-butyl ester N- (cyclohexylacetyl) alanine, iso-butyl ester N- (cyclopentylacetyl) alanine, iso-butyl ester N - [(cyclohex-1-enyl) acetyl] alanine, iso-butyl ester N - [(3-chlorophenyl) acetyl] alanine, 3-methylbut-2-enylthioester N - [(2-phenyl) -2-fluoroacetyl] alanine, ethyl ester N- (3,5-difluorophenylacetyl) phenylglycine methyl ester N- (3,5-difluorophenylacetyl) phenylglycine, iso-butyl ester N- (cyclopentylacetyl) phenylglycine methyl ester N- (cyclopentylacetyl) alanine, methyl ester N- (cyclopropylacetyl) phenylglycine methyl ester N- (cyclopropylacetyl) alanine, methyl ester N - [(3-nitrophenyl) acetyl] methionine, iso-butyl ester. • 4 4 4 4 4 · 4 • 4 ·· · 4 4 4 4 4 4 4 4 4 - 137 24. A compound of formula I XR ³ 4,444,4 for use as a medicament; where in formula I: R ¹ is selected from the group consisting of a) alkyl, alkenyl, alkcycloalkyl, phenyl- (R) _m , -, naphthyl- (R) _m -, wherein R is an alkylene group having 1 to 8 carbon atoms and _m is an integer equal to 0 or 1, cycloalkyl, cycloalkenyl, 3-pyridyl, 4-pyridyl and heteroaryl, other than 3- and 4-pyridyl, of from 3 to 10 atoms and from 1 to 4 heteroatoms selected from the group consisting of oxygen, sulfur and nitrogen, wherein the heteroaryl group is optionally substituted by 1 up to 3 substituents selected from the group consisting of alkyl, alkoxy, aryl, aryloxy, halogen, nitro, thioalkoxy and thioaryloxy, provided that if such heteroaryl contains at least one nitrogen heteroatom, it also contains at least one oxygen and / or sulfur heteroatom; (b) a substituted phenyl group of formula (II): (R) m - in which · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · 138 R is alkylene of 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, cyano, cycloalkyl, halogen, heteroaryl, 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 or 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- (R) _m - substituted in the 5, 6, 7 position and / or 8 with one to four substituents selected from the group consisting of alkyl, alkoxy, halogen, cyano, nitro, trihalomethyl and thioalkoxy, wherein R is C 1 -C 8 alkylene and m is an integer equal to 0 or 1 ; R ² is selected from the group consisting of hydrogen, alkyl, phenyl, alkylalkoxy, alkylthioalkoxy; and R ³ is selected from the group consisting of - (0 ') _n CR ¹⁰ R ⁵ R ⁶ , wherein n is an integer equal to 0, 1 or 2, R ⁵ and R ⁶ are independently selected from the group consisting of hydrogen, alkyl, alkenyl, aryl, heteroaryl, heterocyclic, -NR ⁷ R ⁸ wherein R ⁷ and R ⁸ are independently hydrogen or alkyl and -COOR ⁹ wherein R ⁹ is alkyl and wherein R ⁵ and R ⁶ may furthermore be joined to form a cycloalkyl group, a cycloalkenyl group, an aryl group, a heteroaryl group and a heterocyclic group, and if R ⁵ and R ⁶ are not joined to form an aryl or heteroaryl group, then R ¹⁰ is selected from hydrogen and 139 9 9 9 9 9 9 9 9 9 9 9 9 9 9 99 99 99 99 99 99 99 With the proviso that if n is zero, then R ¹⁰ represents a hydrogen atom and if n is greater than zero and R ⁵ and R ⁶ are joined to form an aryl or heteroaryl group, then R ¹⁰ represents a bond within this group; X represents an oxygen atom or a sulfur atom; 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, and their pharmaceutically acceptable salts, with the additional conditions that: when R ¹ represents phenyl, R ² represents -CH (CH 3) CH ² CH 3, X represents an oxygen atom and X 1 and X represent a hydrogen atom, then R ³ does not represent -CH 2 CH 3 or -CH 2 CH (CH 3) ₂ ; when R ¹ is phenyl, R ³ is -CH 2 CH (CH 3) 2, X is oxygen and X 1 and X are hydrogen, then R ^{2 is} not -CH (CH 3) ₂ when R ¹ is pyrid-3-yl, R ² is ethyl, X is oxygen and X 1 and X are hydrogen, then R ^{3 is} not -CH ₂ CH (CH ₃ ) ₂ when R ¹ is indoxazin-3-yl, 2,4-dimethylthiazole-5- yl, 4-methyl-1,2,5-thiooxadizol-3-yl or 3,5-di (trifluoromethyl) phenyl, R ² is methyl, X is oxygen, and X 'and X are hydrogen, then R ³ does not represent -CH 2 CH (CH 3) 2, and when R ¹ represents -CH 2 -phenyl, R ³ represents -CH ₂ CH ₃ , X represents an oxygen atom and X 'and X "represent a hydrogen atom, then R ² does not represent -CH ₂ CH (CH ₃ ) ₂ . (4) A compound of formula (I) according to claim 24 for use as a medicament, wherein R ¹ represents an unsubstituted phenyl group. A compound of formula I according to claim 24 for use as a medicament, wherein R ¹ represents an unsubstituted naphthyl group selected from the group consisting of 1-naphthyl and 2-naphthyl. - 140 ·· • 0 00 · * 0 ·· • »0 * · · 40 ·· 0 0 0 0 · ··· 0 00 0 0 0 000 0000000 00 0 0 00 00 0 0000 A compound of formula I according to claim 24 for use as a medicament, wherein R ¹ represents a substituted phenyl group of formula II. A compound of formula I according to claim 27 for use as a medicament, wherein the substituted phenyl group is defined as follows: (a) monosubstituted phenyls having a single substitution at the 2, 3 or 4 position, wherein each of the individual substituents is determined by the corresponding R ^a , R ^b and R ^c groups; 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, wherein each of the substituents is determined by the corresponding groups R ¹ , 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. and wherein again each of the substituents is determined by the corresponding groups R ^a , R ^a ', R ^b , R ^b ' and R ^c . A compound of formula I 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 of formula (I) according to claim 24 for use as a medicament, wherein R ¹ represents a phenyl- (R) _m - group, wherein R is alkylene containing from 1 to 8 carbon atoms and _m is 1. 141 A compound of formula I according to claim 30 for use as a medicament, wherein R ¹ is selected from benzyl, 3-phenyln-propyl and 4-phenyl-n-butyl. A compound of formula I according to claim 24 for use as a medicament, wherein R ¹ is selected from the group consisting of alkyl, alkenyl, alkcycloalkyl, cycloalkyl and cycloalkenyl. A compound of formula I according to claim 32 for use as a medicament, wherein R ¹ is alkyl. A compound of formula I according to claim 32 for use as a medicament, wherein R ¹ is cycloalkyl. A compound of formula I according to claim 32 for use as a medicament, wherein R ¹ is alkenyl. A compound of formula I according to claim 32 for use as a medicament, wherein R ¹ is cycloalkenyl. A compound of formula I according to claim 32 for use as a medicament wherein the alkyl, cycloalkyl, alkcycloalkyl, alkenyl and cycloalkenyl groups R ¹ 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. A compound of formula (I) according to claim 24 for use as a medicament, wherein R ¹ is selected from the group consisting of heteroaryl and substituted heteroaryl groups. A compound of formula I according to claim 38 for use as a medicament, wherein the heteroaryl and substituted heteroaryl groups R ¹ are selected from pyrid-3-yl, pyrid-4-yl, · 149 149 149 149 149 149 149 149 149 149 149 149 149 149 149 149 149 149 149 149 149 149 Λ. Thien-2-yl, thien-3-yl, benzothiazol-4-yl, 2-phenylbenzoxazol-5-yl, furan-2-yl, benzofuran-2- yl, benzothiophen-3-yl, 2-chlorothien-5-yl, 3-methylisoxazol-5-yl, 2- (phenylthio) thien-5-yl, 6-methoxythiophen-2-yl, 3-phenyl-1,2,4-thiooxadiazole -5-yl and 2-phenyloxazol-4-yl: A compound of formula (I) according to claim 24 for use as a medicament, wherein R ² is selected from the group consisting of alkyl having from 1 to 4 carbon atoms, alkylalkoxy having from 1 to 4 carbon atoms, phenyl and alkylthioalkoxy having from 1 to 4 carbon atoms. up to 4 carbon atoms. A compound of formula (I) according to claim 30 for use as a medicament, wherein R ² is selected from the group consisting of methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, -CH ₂ CH ₂ SCH ₃ and phenyl. A compound of formula I according to claim 24 for use as a medicament, wherein X 'and X are both hydrogen and X is oxygen. A compound of formula I according to claim 42 for use as a medicament, wherein R ³ is selected from the group consisting of methyl, ethyl, isopropyl, n-propyl, n-butyl, iso-butyl, cyclopentyl, allyl, iso-but -2-enyl, 3-methylpentyl, -CH ₂ -cyclopropyl, -CH ₂ -cyclohexyl, -CH ₂ - (3-tetrahydrofuranyl), -CH ₂ -thien-2-yl, -CH ₂ - (1-methyl) cyclopropyl, -CH ₂ -thien-3-yl, -CH ₂ -C (O) 0 -tert -butyl, -CH ₂ C (CH ₃ ) ₃ , -CH ₂ CH (CH ₂ CH ₃ ) ₂ , -2-methylcyclopentyl, -cyclohex-2-enyl, CH [CH (CH ₃ ) ₂ ] COOCH ₃ , -CH ₂ CH ₂ N (CH ₃ ) ₂ , -CH ₂ C (CH ₃ ) = CH _2, and -CH ₂ CH = C (CH ₃ ) ₂ . A compound of formula I according to claim 24 for use as a medicament, wherein X 'and X are both hydrogen and X is sulfur. - 143 · 143 143 143 143 143 143 143 A compound of formula I according to claim 42 for use as a medicament, wherein R ³ is selected from the group consisting of iso-but-2-enyl and iso-butyl. A compound of formula I according to claim 24 for use as a medicament, wherein the compound of formula I is selected from the group consisting of: N- (phenylacetyl) alanine, iso-butyl ester N- (3-phenylpropionyl) alanine, iso-butyl ester N- (3-methylpentanoyl) alanine, iso-butyl ester N - [(4-chlorophenyl) acetyl] alanine, isobutyl ester N - [(3,4-dichlorophenyl) acetyl] alanine, iso-butyl ester N - [(3-pyridyl) acetyl] alanine, isobutyl ester N - [(1-naphthyl) acetyl] alanine, iso-butyl ester N - [(2-naphthyl) acetyl] alanine, iso-butyl ester N- (4-phenylbutanoyl) alanine, iso-butyl ester N- (5-phenylpentanoyl) alanine, iso-butyl ester N - [(4-pyridyl) acetyl] alanine, isobutyl ester 2 - [(3,4-dichlorophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(3-methoxyphenyl) acetamido] butyric acid, iso-butyl ester 2 - [(4-nitrophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(3,4-methylenedioxyphenyl) acetamido] butyric acid, iso-butyl ester 2 - [(thien-3-yl) acetamido] butyric acid, iso-butyl ester 2 - [(4-chlorophenyl) acetamido] butyric acid, iso - butyl ester 2 - [(3-nitrophenyl) acetamido] butyric acid, iso- Butyl 2 - ((2-hydroxyphenyl) acetamide) butyric acid, iso-butyl ester 2 - [(2-naphthyl) acetamido] butyric acid, iso-butyl ester 2 - [(2,4-dichlorophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(4-bromophenyl) acetamido] butyric acid, iso-butyl ester 0 0 0 0 0 0 < 0 0 3 »000 ·· - 143 • · · 0 0 0 0 2 - [(3-chlorophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(3-fluorophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(benzothiazol-4-yl) acetamido] butyric acid, iso-butyl ester 2 - [(2-methylphenyl) acetamido] butyric acid, iso-butyl ester 2 - [(2-fluorophenyl) acetamido] butyric acid, iso-butyl ester 2 - [(4-fluorophenyl) acetamido] butyric acid, 2 - [(3-bromophenyl) acetamidobutyrate, iso-butyl ester 2 - [( 3-trifluoromethylphenyl) acetamido] butyric acid, iso-butyl ester 2 - [(2-thienyl) acetamido] butyric acid, 2- (phenylacetamido) butyric acid iso-butyl ester, N- (phenylacetyl) valine iso-butyl ester, 2-methylbutyl ester N- (phenylacetyl) methionine, iso-butyl ester N- (phenylacetyl) leucine, iso-butyl ester N - [(3-chlorophenyl) acetyl] alanine, N - [(3-chlorophenyl) acetyl] alanine, N - [(3-chlorophenyl) acetyl] alanine, N - [(3-chlorophenyl) acetyl] alanine, 2-thienylmethyl ester N- [ (3-chlorophenyl) acetyl] alanine, (1-methylcyclopropyl) methyl ester N - [(3-chlorophenyl) acetyl] alanine, 3-thienylmethyl ester N - [(3-chlorophenyl) acetyl] alanine, 2-methylcyclopentyl ester N - [(3-chlorophenyl) acetyl] alanine, 2-methylprop-2-enyl ester N N - [(2-phenylbenzoxazol-5-yl) acetyl] alanine, N - [(3-methylthiophenyl) acetyl] alanine, isobutyl ester, - [(3-chlorophenyl) acetyl] alanine, cyclohex-2-enyl ester N-4 - [(2-furyl) acetyl] alanine, iso-butyl ester N - [(benzofuran-2-yl) acetyl] alanine, iso-butyl ester N - [(benzothiophen-3-yl) acetyl] alanine, iso -butyl ester N - [(2-chloro-5-thienyl) acetyl] alanine, iso -butyl ester N - [(3-methyl-isoxazol-5-yl) acetyl] alanine, isobutyl ester N - [(2-phenylthiothienyl) acetyl] alanine, isobutyl ester N - [(6-methoxybenzothiophen-2-yl) acetyl] alanine, isobutyl ester 145 4 4 4 4 5 6 7 8 9 10 11 12 13 14 ······················ Ν - [(3-phenyl-1,2,4-thiadiazol-5-yl) acetyl] alanine, iso-butyl ester N - [(2-Phenyloxazol-4-yl) acetyl] alanine, iso-butyl ester N - [(3-methylphenyl) acetyl] alanine, iso-butyl ester N - [(2,5-difluorophenyl) acetyl] alanine, iso-butyl ester N - [(3,5-diflurophenyl) acetyl] alanine, isobutyl ester N - [(3-thienyl) acetyl] alanine, iso-butyl ester N - [(4-methylphenyl) acetyl] alanine, isobutyl ester N- (phenylacetyl) alanine, (1-methoxycarbonyl) isobutyl ester N - [(3-nitrophenyl) acetyl] alanine, iso-butyl ester N - [(3,5-difluorophenyl) acetyl] alanine, ethyl ester N - [(3-nitrophenyl) acetyl] methionine, ethyl ester N - [(3-chlorophenyl) acetyl] alanine, isobutyl ester N - [(3-chlorophenyl) acetyl] alanine, 2- (N, N-dimethylamino) ethyl ester 2 - [(3,5-dichlorophenyl) acetamido] hexanoic acid methyl ester N - [(3,5-dichlorophenyl) acetyl] alanine, iso-butyl ester N- (cyclohexylacetyl) alanine, iso-butyl ester N- (cyclopentylacetyl) alanine, iso-butyl ester N - [(cyclohex-1-enyl) acetyl] alanine, iso-butyl ester N - [(3-chlorophenyl) acetyl] alanine, 3-methylbut-2-enylthioester N - [(2-phenyl) -2-fluoroacetyl] alanine, ethyl ester N- (3,5-difluorophenylacetyl) phenylglycine methyl ester N- (3,5-difluorophenylacetyl) phenylglycine, iso-butyl ester N- (cyclopentylacetyl) phenylglycine methyl ester N- (cyclopentylacetyl) alanine, methyl ester N- (cyclopropylacetyl) phenylglycine methyl ester N- (cyclopropylacetyl) alanine, methyl ester N - [(3-nitrophenyl) acetyl] methionine, iso-butyl ester A pharmaceutical composition which is pharmaceutically active A pharmaceutical composition, a pharmaceutically inert carrier of a compound of Formula I: contains quantity in which R ¹ is selected from the group consisting of a) alkyl, alkenyl, alkcycloalkyl, phenyl- (R) _m , -, naphthyl- (R) _m -, wherein R is an alkylene group having 1 to 8 carbon atoms and _m is an integer equal to 0 or 1, cycloalkyl, cycloalkenyl, 3-pyridyl, 4-pyridyl and heteroaryl, other than 3- and 4-pyridyl, of from 3 to 10 atoms and from 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen, wherein the heteroaryl group is optionally substituted by 1 up to 3 substituents selected from the group consisting of alkyl, alkoxy, aryl, aryloxy, halogen, nitro, thioalkoxy and thioaryloxy, provided that if such heteroaryl contains at least one nitrogen heteroatom, it also contains at least one oxygen and / or sulfur heteroatom; (b) a substituted phenyl group of formula (II): (R) _m - in which 147 ΦΦΦΦ · · · · · · φ φ φ φ φ φ φ φ φ R is alkylene of 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, cyano, cycloalkyl, halogen, heteroaryl, 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 or 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- (R) _m - substituted in the 5, 6, 7 position and / or 8 with one to four substituents selected from the group consisting of alkyl, alkoxy, halogen, cyano, nitro, trihalomethyl and thioalkoxy, wherein R is C 1 -C 8 alkylene and m is an integer equal to 0 or 1 ; R ² is selected from the group consisting of hydrogen, alkyl, phenyl, alkylalkoxy, alkylthioalkoxy; and R ³ is selected from the group consisting of - (CH ₂ ) _n CR ¹⁰ R ⁵ R ⁶ , wherein n is an integer equal to 0, 1 or 2, R ⁵ and R ⁶ are independently selected from the group consisting of hydrogen , alkyl, alkenyl, aryl, heteroaryl, heterocyclic, -NR ⁷ R ⁸ wherein R ⁷ and R ⁸ are independently hydrogen or alkyl and -COOR ⁹ wherein R ⁹ is alkyl and wherein R ⁵ and R ^{6 are} furthermore they may be linked to form a cycloalkyl group, a cycloalkenyl group, an aryl group, a heteroaryl group and a heterocyclic group, and if R ⁵ and R ⁶ are not joined to form an aryl or heteroaryl group, R ¹⁰ is selected from the group consisting of hydrogen and • · 4 I · 4 With the proviso that if n is zero, then R ¹⁰ represents a hydrogen atom and if n is greater than zero and R ⁵ and R ⁶ are joined to form an aryl or heteroaryl group, then R ¹⁰ represents a bond within this group; X represents an oxygen atom or a sulfur atom; 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, and their pharmaceutically acceptable salts, with the additional conditions that: when R ¹ represents phenyl, R ² represents -CH (CH 3) CH ² CH 3, X represents an oxygen atom and X 1 and X represent a hydrogen atom, then R ³ does not represent -CH ₂ CH ₃ or -CH ₂ CH (CH ₃ ) ₂ ; when R ¹ is phenyl, R ³ is -CH 2 CH (CH 3) 2, X is oxygen and X ¹ and X are hydrogen, then R ^{2 is} not -CH (CH 3) ₂ when R ¹ is pyrid-3-yl, R ² is ethyl, X is oxygen and X 1 and X are hydrogen, then R ^{3 is} not -CH ₂ CH (CH ₃ ) ₂ when R ¹ is indoxazin-3-yl, 2,4-dimethylthiazole-5- yl, 4-methyl-1,2,5-thiooxadizol-3-yl or 3,5-di (trifluoromethyl) phenyl, R ² is methyl, X is oxygen, and X 'and X are hydrogen, then R ³ does not represent -CH 2 CH (CH 3) 2, and when R ¹ represents -CH 2 -phenyl, R ³ represents -CH ₂ CH ₃ , X represents an oxygen atom and X 1 and X represent a hydrogen atom, then R ² does not represent -CH ₂ CH (CH ₃ ) ₂ . 48. A pharmaceutical composition according to claim 47, characterized in that R ¹ represents an unsubstituted phenyl group. 49. The pharmaceutical composition according to claim 48, characterized in that R ¹ is an unsubstituted naphthyl group selected from the group consisting of 1-naphthyl and 2-naphthyl. 149 50. The pharmaceutical composition according to claim 47, characterized in that R ¹ represents a substituted phenyl group of formula II. 51. The pharmaceutical composition of claim 50 wherein the substituted phenyl group is defined as follows: (a) monosubstituted phenyls having a single substitution at the 2, 3 or 4 position, wherein each of the individual substituents is determined by the corresponding R ^a , R ^b and R ^c groups; 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 positions 3,5 or at positions 3,6, wherein each of the substituents is determined by the corresponding groups R ¹ , R ^a ', R ^b , R ^b ' and (c) trisubstituted phenyls having three substituents at the 2-position, 3,4, at positions 2,3,5, at positions 2,3,6, at positions 3,4,5, and at positions 3,4,6 and wherein, in turn, each of the substituents is determined by the corresponding groups R ^a , R a ^and ', R ^b, R ^b' and R ^c 52. The pharmaceutical composition of claim 51 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 pharmaceutical composition according to claim 47, wherein R ¹ represents a phenyl- (R) _m - group, wherein R is alkylene containing from 1 to 8 carbon atoms and _m is 1. ·· ♦ · - 150 54. The pharmaceutical composition according to claim 53, characterized in that R ¹ is selected from the group consisting of benzyl, 3-phenyl-n-propyl and 4-phenyl-n-butyl. 55. The pharmaceutical composition according to claim 47, characterized in that R ¹ is selected from the group consisting of alkyl, alkenyl, alkcycloalkyl, cycloalkyl and cycloalkenyl. 56. Pharmaceutical that R ¹ represents composition alkyl. according to claim 55, characterized se by 57. Pharmaceutical that R ¹ represents composition cycloalkyl according to • claim 55, characterized se by 58. Pharmaceutical that R ¹ represents composition alkenyl. according to claim 55, characterized se by 59. Pharmaceutical that R ¹ represents composition according to cycloalkenyl. claim 55, characterized se by 60. Pharmaceutical composition according to claim 55, characterized se by wherein the alkyl, cycloalkyl, alkcycloalkyl, alkenyl and cycloalkenyl groups R ¹ 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. 61. The pharmaceutical composition according to claim 47, characterized in that R ¹ is selected from the group consisting of heteroaryl and substituted heteroaryl groups. 62. The pharmaceutical composition of claim 61 wherein the heteroaryl and substituted heteroaryl groups R &lt; ¹ &gt; are selected from pyrid-3-yl, pyrid-4-yl, thien-2- 151. 2 - [(3,5-dichlorophenyl) acetamido] hexanoic acid methyl ester N - [(3,5-dichlorophenyl) acetyl] alanine, isobutyl ester N- (cyclohexylacetyl) alanine, iso-butyl ester N- (cyclopentylacetyl) alanine, iso-butyl ester N - [(cyclohex-1-enyl) acetyl] alanine, iso-butyl ester N - [(3-chlorophenyl) acetyl] alanine, 3-methylbut-2-enylthioester N - [(2-phenyl) -2-fluoroacetyl] alanine, ethyl ester N- (3,5-difluorophenylacetyl) phenylglycine methyl ester N- (3,5-difluorophenylacetyl) phenylglycine, iso-butyl ester N- (cyclopentylacetyl) phenylglycine methyl ester N- (cyclopentylacetyl) alanine, methyl ester N- (cyclopropylacetyl) phenylglycine methyl ester N- (cyclopropylacetyl) alanine, methyl ester N - [(3-nitrophenyl) acetyl] methionine, iso-butyl ester.