CN101817769B

CN101817769B - Carbamido peptide aminopeptidase N inhibitor and application thereof

Info

Publication number: CN101817769B
Application number: CN 201010146905
Authority: CN
Inventors: 徐文方; 宿莉; 方浩
Original assignee: Shandong University
Current assignee: Shandong University
Priority date: 2010-04-15
Filing date: 2010-04-15
Publication date: 2013-09-11
Anticipated expiration: 2030-04-15
Also published as: CN101817769A

Abstract

The invention provides a carbamido peptide aminopeptidase N inhibitor and application thereof. The invention provides the superactive peptide aminopeptidase N inhibitor, thereby being capable of curing the disease that the activity or the express of the aminopeptidase N is abnormal. Specifically, the invention relates to a peptide compound with the structures of general formulas (I), (II) or (III), and further relates to various optical isomers thereof, a pharmaceutically acceptable salt, a solvate and a prodrug. The invention further relates to a drug composite of the peptide compound which comprises the structures of the general formulas (I), (II) or (III) and use therefore for preparing the drug.

Description

Urea-based peptide aminopeptidase N inhibitor and application thereof

Technical Field

The invention relates to a carbamido peptide aminopeptidase N inhibitor, a preparation method and application thereof, belonging to the technical field of chemistry.

Background

Aminopeptidase N (APN, CD13) is a family of type II membrane-bound glycoproteins with molecular weights of about 150Kd, belonging to the Gluzincins subfamily of the zinc ion-dependent metalloprotease and aminopeptidase M1 family, present in the cell membrane as homodimers and involved in the degradation of the N-terminal amino acids of the substrate. Aminopeptidase N is widely distributed in GM-SCF and myeloid cells at various stages of development; in non-blood cells, stromal cells distributed in hematopoietic organs, tumor vascular endothelial cells, various epithelial cells including renal proximal convoluted tubule epithelial cells, bile duct epithelial cells, small intestine brush border epithelial cells and the like; also distributed on the surface of fibroblasts, mesenchymal tumor cells, etc.; a small amount of soluble APN was also present in the serum. APN participates in physiological regulation of organisms and plays an important role in a plurality of physiological and pathological regulation networks such as blood cell differentiation, angiogenesis, cell proliferation and apoptosis, immunoregulation and the like. Research has shown that aminopeptidase N plays an important role in tumorigenesis, development, invasion and metastasis, apoptosis, tumor angiogenesis and viral infection.

1) Aminopeptidase N is expressed at high levels on the surface of tumor cells. The enzyme can degrade extracellular matrix, thereby promoting tumor cell invasion and metastasis. Extracellular matrix plays a very important role in maintaining the stability of cell junctions and intercellular signaling. Degradation of the extracellular matrix can also promote growth and proliferation of tumors by promoting the release of growth factors present therein (Sato Y, biol. pharm. Bull., 2004, 27 (6): 772-) 776; Saiki, I.; et al. int. J. cancer., 1993, 54, 137; Menrad A., Speicher D., Wacker J., et al. cancer Res., 1993, 53 (6): 1450-) 1455). 2) Aminopeptidase N can stimulate vascular endothelial cells to release tumor microvascular formation related factors and promote angiogenesis of tumor cells. The APN is highly expressed in endothelial cells and sub-endothelial cells of new vessels, and can facilitate the endothelial cells to invade other surrounding tissues. This is also the basis for angiogenesis, which is the first step in tumor growth and metastasis. 3) APN is expressed on the surfaces of granulocytes and lymphocytes in a large quantity, and is also involved in T lymphocyte-dependent inflammatory reaction; can also be expressed on the surface of antigen presenting cells to degrade immune active substances (such as interleukin-8); the recognition of T cells which are involved in antigen processing and cell surface major histocompatibility complex II (MHC-II) adhesion antigenic determinant dependence on the antigen reduces the recognition capability of the T cells to the antigen, simultaneously weakens the recognition and killing capability of macrophages and NK cells to tumor cells, reduces the immunity of the organism, thereby promoting the proliferation of the tumor cells and inhibiting the apoptosis of the tumor cells. 4) Aminopeptidase N acts as a receptor on the surface of human coronavirus HCoV-229E and transmissible gastroenteritis virus (TGEV) and infects the upper respiratory tract (e.g.: SARS) and acute enteritis, and its role is related to the activity of enzymes (Delmas, b., et al. nature, 1992, 357, 417; yeager, c.l.; et al. nature, 1992, 357, 420). APN is also involved in T lymphocyte-dependent inflammatory responses and in the entry of HIV viral particles into host cells. Studies have shown that aminopeptidase N activity in HIV-infected patients is much higher than in healthy volunteers (Shen W, Li B, et al. blood, 2000, 96(8), 2887; Shipp MA, et al. blood, 1993, 82(4), 1052). 5) Aminopeptidase N is involved in the degradation of the endogenous analgesic substances endorphins and enkephalins, causing the excessive release of substance P, resulting in pain. 6) Aminopeptidase N degrades angiotensin, and is involved in the regulation of body blood pressure (Mitsui, t.; et al biol pharm bull, 2004, 27, 768).

For over a decade, research and development of APN inhibitors has been extremely rapid, but to date there has been only one drug on the market, ubenimex. APN inhibitors act as inhibitors of peptidases, mostly peptides or peptide analogs, and are susceptible to degradation by enzymes in the body; in addition, because the control network for controlling tumor growth, proliferation and apoptosis is very large and complex, for example, the regulation network can complementarily promote tumor growth, proliferation and inhibit apoptosis through in vivo signal transduction of various growth factor receptor tyrosine kinases or through activation of integrin receptors, which is the reason why most of non-cytotoxic target type antitumor drugs including APN inhibitors are killed by gun in clinical stage. In addition, most of aminopeptidase N inhibitors currently under clinical or preclinical study are natural products, for example, Ubenimex (Ubenimex) is a dipeptide-like structure containing a beta-amino acid, and is currently used as an immunopotentiator for treating leukemia, is separated from a culture solution of Streptomyces olivaceus (Streptomyces olivoreticuli), and is expensive in total synthesis and limited in source.

The ureido peptide compound designed by the invention is found in the screening aiming at the aminopeptidase N activity, and the activity of several drug molecules is similar to or superior to that of the currently unique ubenimex on the market.

Disclosure of Invention

Aiming at the defects of the prior art, the invention provides a carbamido peptide aminopeptidase N inhibitor, a preparation method and application thereof.

The technical scheme of the invention is as follows:

peptoid compounds having the general formula (I), (II) or (III), as well as optical isomers, diastereomers and racemic mixtures thereof, pharmaceutically acceptable salts, solvates or prodrugs thereof:

wherein,

R₁is hydrogen, a substituent on the alpha carbon of various natural or unnatural amino acids, including: various α -amino acids such as glycine, alanine, valine, phenylalanine, isoleucine, leucine, methionine, lysine, ornithine, aspartic acid, asparagine, glutamic acid, glutamine, serine, threonine, histidine, tryptophan, arginine, cysteine, citrulline, proline, hydroxyproline, tyrosine, and the like; various beta-amino acids such as beta-aminopropionic acid, beta-aminophenylpropionic acid, and the like; gamma-amino acids such as gamma-aminobutyric acid and the like, D-amino acids such as glycine, alanine, valine, phenylalanine, isoleucine, leucine, methionine, lysine, ornithine, aspartic acid, asparagine, glutamic acid, glutamine, serine, threonine, histidine, tryptophan, arginine, cysteine, citrulline, proline, hydroxyproline, tyrosine and the like in D form; other amino acids such as various delta-amino acids, and the like.

R₁' is hydrogen, a substituent on the alpha carbon of various natural or unnatural amino acids, including: various α -amino acids such as glycine, alanine, valine, phenylalanine, isoleucine, leucine, methionine, lysine, ornithine, aspartic acid, asparagine, glutamic acid, glutamine, serine, threonine, histidine, tryptophan, arginine, cysteine, citrulline, proline, hydroxyproline, tyrosine, and the like; various beta-amino acids such as beta-aminopropionic acid, beta-aminophenylpropionic acid, and the like; gamma-amino acids such as gamma-aminobutyric acid, etc., and D-amino acids such as D-form glycine, alanine, valine, phenylalanine, isoleucine, leucine, methionine, lysine, ornithine, aspartic acid, asparagine, glutamic acid, glutamine, serine, threonine, histidine, tryptophan, arginine, cysteine, citrulline, proline, hydroxyproline, etcAmino acids, amino acids; other amino acids such as various delta-amino acids, and the like.

R₂Is hydrogen, C1-12 aliphatic alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl; optionally substituted with one or more of the following groups: hydroxy, halogen, nitro, cyano, guanidino, carboxy, haloC 1-12 alkyl, C1-12 alkoxy, C1-12 alkyl, C1-12 cycloalkyl, aryl, heteroaryl, or arylC 1-12 alkyl;

x is hydroxamic acid, carboxyl; or C1-12 aliphatic alkoxy, aryloxy, arylalkoxy, heteroaryloxy, heteroarylalkoxy, C1-12 aliphatic alkylamino, arylamino, arylalkylamino, heteroarylamino; optionally substituted with one or more of the following groups: hydroxy, halogen, nitro, cyano, guanidino, carboxy, haloC 1-12 alkyl, C1-12 alkoxy, C1-12 alkyl, C1-12 cycloalkyl, aryl, heteroaryl, arylC 1-12 alkyl;

y is hydrogen, aroyl, heteroaroyl, aryl C1-6 alkanoyl, heteroaryl C1-9 alkanoyl, C1-6 alkanoyl, arylsulfonyl, heterosulfonyl, aryl C1-6 alkylsulfonyl or heteroaryl C1-9 alkylsulfonyl; various protecting groups for protecting amino acid amino group, such as t-butoxycarbonyl, benzyloxycarbonyl, fluorenylmethyloxycarbonyl, 2-biphenyl-2-propoxycarbonyl, phthalimido, p-toluenesulfonyl, trityl, formyl, acetyl or trifluoroacetyl group and the like.

The carbon represented by the above formula (I) or (II) has S configuration.

Preferably, the ureido peptoid compounds (I), (II) or (III) include the following compounds:

preferably, the ureido peptoid compound (I) is one of the following compounds:

1 a: 2- (3-tert-butylureido) -N-hydroxyacetamide,

2 a: (2S) -2-methyl-2- (3-tert-butylureido) -N-hydroxyacetamide,

3 a: (2S) -2-isopropyl-2- (3-tert-butylureido) -N-hydroxyacetamide,

4 a: (2S) -2- (2-methyl) -propyl-2- (3-tert-butylureido) -N-hydroxyacetamide,

5 a: (2S) -2- (1-methyl) -propyl-2- (3-tert-butylureido) -N-hydroxyacetamide,

6 a: (2S) -2-benzyl-2- (3-tert-butylureido) -N-hydroxyacetamide,

7 a: (2S) -2- (methylthio) -ethyl-2- (3-tert-butylureido) -N-hydroxyacetamide,

8 a: (2S) -2- [4- (N-benzyloxycarbonyl) -aminobutyl ] -2- (3-tert-butylureido) -N-hydroxyacetamide,

1 b: 2- (3-benzylureido) -N-hydroxyacetamide,

2 b: (2S) -2-methyl-2- (3-benzylureido) -N-hydroxyacetamide,

3 b: (2S) -2-isopropyl-2- (3-benzylureido) -N-hydroxyacetamide,

4 b: (2S) -2- (2-methyl) -propyl-2- (3-benzylureido) -N-hydroxyacetamide,

5 b: (2S) -2- (1-methyl) -propyl-2- (3-benzylureido) -N-hydroxyacetamide,

6 b: (2S) -2-benzyl-2- (3-benzylureido) -N-hydroxyacetamide,

7 b: (2S) -2- (methylthio) -ethyl-2- (3-benzyl-tert-butylureido) -N-hydroxyacetamide,

8 b: (2S) -2- [4- (N-benzyloxycarbonyl) -aminobutyl ] -2- (3-benzyl-tert-butylureido) -N-hydroxyacetamide,

1 c: 2- (3-phenethylureido) -N-hydroxyacetamide,

2 c: (2S) -2-methyl-2- (3-phenethylureido) -N-hydroxyacetamide,

3 c: (2S) -2-isopropyl-2- (3-phenethylureido) -N-hydroxyacetamide,

4 c: (2S) -2- (2-methyl) -propyl-2- (3-phenethylureido) -N-hydroxyacetamide,

5 c: (2S) -2- (1-methyl) -propyl-2- (3-phenethylureido) -N-hydroxyacetamide,

6 c: (2S) -2-benzyl-2- (3-phenethylureido) -N-hydroxyacetamide,

7 c: (2S) -2- (methylthio) -ethyl-2- (3-phenethylureido) -N-hydroxyacetamide or

8 c: (2S) -2- [4- (N-benzyloxycarbonyl) -aminobutyl ] -2- (3-phenethylureido) -N-hydroxyacetamide.

Preferably, the ureido peptoid compound (II) is one of the following compounds:

1 f: 2- [3- (1-hydroxycarbamoyl-3-methyl-butyl) -ureido ] -4-methylpentanoic acid,

2 f: 2- [3- (1-hydroxycarbamoyl-3-methyl-butyl) -ureido ] -3-phenylpropionic acid,

3 f: 2- [3- (1-hydroxycarbamoyl-2-phenyl-ethyl) -ureido ] -4-methylpentanoic acid,

4 f: 2- [3- (1-hydroxycarbamoyl-2-phenyl-ethyl) -ureido ] -3-phenylpropionic acid,

1 g: 2- [3- (1-hydroxycarbamoyl-3-methyl-butyl) -ureido ] -4-methylpentanoic acid hydroxyamide,

2 g: 2- [3- (1-hydroxycarbamoyl-3-methyl-butyl) -ureido ] -3-phenylpropionic acid hydroxyamide,

3 g: 2- [3- (1-hydroxycarbamoyl-2-phenyl-ethyl) -ureido ] -4-methylpentanoic acid hydroxyamide or

4 g: 2- [3- (1-hydroxycarbamoyl-2-phenyl-ethyl) -ureido ] -3-phenylpropionic acid hydroxyamide.

Preferably, the ureido peptoid compound (III) is one of the following compounds:

1 d: 6-benzyloxycarbonylamino-2- (3-carboxymethylureido) -hexanoic acid,

2 d: (2S, 2' S) -6-benzyloxycarbonylamino-2- [3- (2-methyl) -carboxymethylureido ] -hexanoic acid,

3 d: (2S, 1' S) -6-benzyloxycarbonylamino-2- [3- (1-carboxy-ethyl) -ureido ] -hexanoic acid,

4 d: (2S, 1' S) -6-benzyloxycarbonylamino-2- [3- (1-carboxy-2-methylpropyl) -ureido ] -hexanoic acid,

5 d: (2S, 1' S) -6-benzyloxycarbonylamino-2- [3- (1-carboxy-3-methylbutyl) -ureido ] -hexanoic acid,

6 d: (2S, 1' S) -6-benzyloxycarbonylamino-2- [3- (1-carboxy-2-methylbutyl) -ureido ] -hexanoic acid,

7 d: (2S, 1' S) -6-benzyloxycarbonylamino-2- [3- (1-carboxy-2-benzyl) -ureido ] -hexanoic acid,

8 d: (2S, 1' S) -6-benzyloxycarbonylamino-2- [3- (1-carboxy-3-methylthiopropyl) -ureido ] -hexanoic acid,

1 e: (2S, 1' S) -6-benzyloxycarbonylamino-2- [3- (5-benzyloxycarbonylamino-1-carboxy-pentyl) -ureido ] -hexanoic acid,

2 e: (3' S) - [ 5-Hydroxyaminocarbonyl-5- (3-Hydroxyaminocarbonylmethyl-ureido) -pentyl ] -carbamic acid benzyl ester,

3 e: (3 'S, 1' S) - { 5-Hydroxyaminocarbonyl-5- [3- (1-Hydroxyaminocarbonylethyl) -ureido ] -pentyl } -carbamic acid benzyl ester,

4 e: (3 'S, 1' S) - { 5-Hydroxyaminocarbonylmethyl-5- [3- (1-Hydroxyaminocarbonyl-2-methyl) -propylureido ] -pentyl } -carbamic acid benzyl ester,

5 e: (3 'S, 1' S) - { 5-Hydroxyaminocarbonylmethyl-5- [3- (1-Hydroxyaminocarbonyl-3-methyl) -butylureido ] -pentyl } -carbamic acid benzyl ester,

6 e: (3 'S, 1' S) - { 5-Hydroxyaminocarbonylmethyl-5- [3- (1-Hydroxyaminocarbonyl-2-phenethylureido ] -pentyl } -carbamic acid benzyl ester,

7 e: (3 'S, 1' S) - { 5-Hydroxyaminocarbonylmethyl-5- [3- (1-Hydroxyaminocarbonyl-3-methylsulfanyl) -propylureido ] -pentyl } -carbamic acid benzyl ester,

8 e: (3 'S, 1' S) - {5- [3- (5-benzyloxycarbonylamino-1-hydroxyaminocarbonyl-pentyl) -ureido ] -5-hydroxyaminocarbonyl-pentyl } -carbamic acid benzyl ester,

1 h: (2S) -6-benzyloxycarbonyl-2- (3-methoxycarbonylmethyl-ureido) hexanoic acid methyl ester,

2 h: (2S, 2' S) -6-benzyloxycarbonyl 2- [3-2- (methoxycarbonylethyl) -ureido ] hexanoic acid methyl ester,

3 h: (2S, 1' S) -6-benzyloxycarbonyl-2- [3- (1-methoxycarbonyl-2-methylpropyl) -ureido ] hexanoic acid methyl ester,

4 h: (2S, 1' S) -6-benzyloxycarbonyl-2- [3- (1-methoxycarbonyl-3-methylbutyl) -ureido ] hexanoic acid methyl ester,

5 h: (2S, 1' S) -6-benzyloxycarbonyl-2- [3- (1-methoxycarbonyl-2-methylbutyl) -ureido ] hexanoic acid methyl ester,

6 h: (2S, 1' S) -6-benzyloxycarbonyl-2- [3- (1-methoxycarbonyl-2-phenylethyl) -ureido ] hexanoic acid methyl ester,

7 h: (2S, 1' S) -6-benzyloxycarbonyl-2- [3- (1-methoxycarbonyl-3-methylthiopropyl) -ureido ] hexanoic acid methyl ester,

1 i: (2S) -6-benzyloxycarbonyl-2- (3-benzyloxycarbonylmethyl-ureido) hexanoic acid benzyl ester,

2 i: (2S, 2' S) -6-benzyloxycarbonyl 2- [3-2- (benzyloxycarbonylethyl) -ureido ] hexanoic acid benzyl ester,

3 i: (2S, 1' S) -6-benzyloxycarbonyl-2- [3- (1-benzyloxycarbonyl-2-methylpropyl) -ureido ] hexanoic acid benzyl ester,

4 i: (2S, 1' S) -6-benzyloxycarbonyl-2- [3- (1-benzyloxycarbonyl-3-methylbutyl) -ureido ] hexanoic acid benzyl ester,

5 i: (2S, 1' S) -6-benzyloxycarbonyl-2- [3- (1-benzyloxycarbonyl-2-methylbutyl) -ureido ] hexanoic acid benzyl ester,

6 i: (2S, 1' S) -6-benzyloxycarbonyl-2- [3- (1-benzyloxycarbonyl-2-phenylethyl) -ureido ] hexanoic acid benzyl ester or

7 i: benzyl (2S, 1' S) -6-benzyloxycarbonyl-2- [3- (1-benzyloxycarbonyl-3-methylthiopropyl) -ureido ] hexanoate.

Among the above compounds, the most preferred are: 4b, 5b, 6b, 7b, 4c, 6c, 4d, 4 e.

In addition, the present invention also includes a pharmaceutical composition suitable for oral administration to a mammal comprising a peptoid compound of the above general formula (I), (II) or (III) and a pharmaceutically acceptable carrier, optionally comprising one or more pharmaceutically acceptable excipients.

In addition, the present invention also includes a pharmaceutical composition suitable for parenteral administration to a mammal comprising a peptoid compound of the above general formula (I), (II) or (III) and a pharmaceutically acceptable carrier, optionally comprising one or more pharmaceutically acceptable excipients.

The use of these peptoid compounds in medicaments for the prevention or treatment of mammalian diseases associated with abnormal aminopeptidase N activity or expression. The related mammal diseases with abnormal aminopeptidase N activity or expression comprise: inflammation, cancer, multiple sclerosis, various tissue ulcers or ulcerative conditions, periodontal disease, epidermolysis bullosa, leukemia, and the like. Therefore, the invention also relates to a pharmaceutical composition containing the compound with the structure (I), (II) or (III).

Detailed Description

Definitions and terms used

The terms and definitions used herein have the following meanings:

"substituents on the alpha carbon of various natural or unnatural amino acids": refers to 20 kinds of natural amino acids, non-natural alpha-amino acids, beta-amino acids, gamma-amino acids, delta-amino acids, D-amino acids, etc., and their derivatives, preferably natural alpha-amino acids, such as phenylalanine, isoleucine, leucine, methionine.

"halo", or "halogen" includes fluorine, chlorine, bromine or iodine.

"cycloalkyl" is a substituted or unsubstituted, saturated or unsaturated, cyclic alkyl group containing carbon atoms and/or one or more heteroatoms. The rings may be monocyclic or fused, bridged or spiro ring systems. Monocyclic rings usually have 3 to 9 atoms, preferably 4 to 7 atoms, and polycyclic rings have 7 to 17 atoms, preferably 7 to 13 atoms.

"heteroalkyl" refers to a saturated or unsaturated chain containing carbon atoms and at least one heteroatom, any one of which is not adjacent. The heteroalkyl group contains 2 to 15 atoms (carbon atoms), preferably 2 to 10 atoms. Heteroalkyl groups may be straight or branched, substituted or unsubstituted.

"aryl" means an aromatic carbocyclic group, preferably an aromatic ring containing 6 to 10 carbon atoms.

"heteroaryl" is an aromatic heterocycle, which may be a monocyclic or bicyclic group.

"cycloalkoxy" is a substituted or unsubstituted, saturated or unsaturated, cyclic alkoxy group containing carbon atoms and/or one or more heteroatoms. The rings may be monocyclic or fused, bridged or spiro ring systems. Monocyclic rings usually have 3 to 9 atoms, preferably 4 to 7 atoms, and polycyclic rings have 7 to 17 atoms, preferably 7 to 13 atoms.

"cycloalkylamino" is a substituted or unsubstituted, saturated or unsaturated, cyclic alkylamino radical containing carbon atoms and/or one or more heteroatoms. The rings may be monocyclic or fused, bridged or spiro ring systems. Monocyclic rings usually have 3 to 9 atoms, preferably 4 to 7 atoms, and polycyclic rings have 7 to 17 atoms, preferably 7 to 13 atoms.

"Heteroalkoxy" refers to a saturated or unsaturated chain containing carbon atoms and at least one heteroatom, any one of which is not adjacent. The heteroalkoxy group contains 2 to 15 atoms (carbon atoms), preferably 2 to 10 atoms. The heteroalkoxy group can be straight or branched, substituted or unsubstituted.

"Heteroalkylamino" refers to a saturated or unsaturated chain containing carbon atoms and at least one heteroatom, any one of which is not adjacent. The heteroalkylamine group contains 2 to 15 atoms (carbon atoms), preferably 2 to 10 atoms. The heteroalkylamine group can be straight-chain or branched, substituted or unsubstituted.

"aryloxy" refers to an aromatic carbocyclic oxy group, the preferred aromatic ring containing 6 to 10 carbon atoms.

"arylamino" refers to an aromatic carbocyclic amine group, the preferred aromatic ring containing 6 to 10 carbon atoms.

A "heteroaryloxy" group is an aromatic heterocycloxy group, which may be a monocyclic or bicyclic group.

A "heteroarylamine" is an aromatic heterocyclic amine group that may be a monocyclic or bicyclic group.

"aroyl" refers to a group having a carbonyl group attached to the end of an aromatic carbocyclic ring. Preferred aromatic rings contain 6 to 10 carbon atoms.

"Cycloalkanoyl" is a substituted or unsubstituted, saturated or unsaturated cyclic terminal carbonyl-containing group containing carbon atoms and/or one or more heteroatoms. The rings may be monocyclic or fused, bridged or spiro ring systems. Monocyclic rings usually have 3 to 9 atoms, preferably 4 to 7 atoms, and polycyclic rings have 7 to 17 atoms, preferably 7 to 13 atoms.

"Heteroaroyl" is a group having a carbonyl group attached to the end of an aromatic heterocycle, and may be a monocyclic or bicyclic group. Preferred heteroaryl groups include, for example, thienyl, furyl, pyrrolyl, pyridyl, pyrazinyl, thiazolyl, pyrimidinyl, quinolinyl, and tetrazolyl, benzothiazolyl, benzofuryl, indolyl, and the like.

"aryl" refers to an aromatic carbocyclic group. Preferred aromatic rings contain 6 to 10 carbon atoms.

"cycloalkyl" is a substituted or unsubstituted, saturated or unsaturated cyclic group containing carbon atoms and/or one or more heteroatoms. The rings may be monocyclic or fused, bridged or spiro ring systems. Monocyclic rings usually have 3 to 9 atoms, preferably 4 to 7 atoms, and polycyclic rings have 7 to 17 atoms, preferably 7 to 13 atoms.

By "pharmaceutically acceptable salt" is meant a therapeutically effective and non-toxic salt form of a compound of formula (I), (II) or (III). Which can form a cationic salt from any basic group, such as an amino group. Many such salts are known in the art. A cationic salt formed on any acidic group (e.g., a carboxyl group), or an anionic salt formed on any basic group (e.g., an amino group). Many of these salts are known in the art, such as cationic salts including salts of alkali metals (e.g., sodium and potassium) and alkaline earth metals (e.g., magnesium and calcium) and organic salts (e.g., amine salts). These salts are well known to the skilled artisan and the skilled artisan can prepare any of the salts provided by the knowledge in the art. In addition, the skilled artisan may select one salt and select another salt depending on solubility, stability, ease of formulation, etc. The determination and optimization of these salts is within the experience of the skilled artisan.

A "solvate" is a complex of a solute (e.g., an aminopeptidase N inhibitor) and a solvent (e.g., water). See J.Honig et al, The Van Nostrand and chemistry's Dictionary, p.650 (1953). Pharmaceutically acceptable solvents useful in the present invention include those that do not interfere with the biological activity of the aminopeptidase N inhibitor (e.g., water, ethanol, acetic acid, N-dimethylformamide, dimethylsulfoxide, and solvents known or readily determined by those skilled in the art).

The terms "optical isomers", "enantiomers", "diastereomers", "racemates" and the like, as used herein, define all possible stereoisomeric forms of the compounds of the present invention or physiological derivatives thereof. Unless otherwise indicated, the chemical designation of the compounds of the invention includes mixtures of all possible stereochemical forms, which mixtures comprise all diastereomers and enantiomers of the basic structural molecule, as well as the substantially pure individual isomeric forms of the compounds of the invention, i.e. containing less than 10%, preferably less than 5%, in particular less than 2%, most preferably less than 1% of other isomers. Various stereoisomeric forms of the peptoid compounds of the present invention are expressly included within the scope of the present invention.

The peptoid compounds of formula (I), (II) or (III) may also exist in other protected forms or derivatives, which are obvious to those skilled in the art and are intended to be included within the scope of the present invention.

The substituents described above may themselves be substituted by one or more substituents. Such substituents include those listed in C.Hansch and A.Leo, scientific Constants for Correlation Analysis in Chemistry and Biology (1979). Preferred substituents include, for example, alkyl, alkenyl, alkoxy, hydroxy, oxy, nitro, amino, aminoalkyl (e.g., aminomethyl, and the like), cyano, halo, carboxy, carbonylalkoxy (e.g., carbonylathoxy, and the like), thio, aryl, cycloalkyl, heteroaryl, heterocycloalkyl (e.g., piperidinyl, morpholinyl, pyrrolyl, and the like), imino, hydroxyalkyl, aryloxy, arylalkyl, and combinations thereof.

The preparation method of the compound comprises the following reaction steps and the reaction formula:

the preparation of compound (I) is as follows:

using optical amino acid as a raw material, sequentially protecting carboxyl through esterification, carrying out condensation connection on corresponding amine through triphosgene, and carrying out corresponding reaction on methyl ester to obtain a target compound; or the corresponding amine is converted into isocyanate by triphosgene, and then condensed with methyl ester of optical amino acid, and then the methyl ester is subjected to corresponding reaction to obtain the target compound.

The synthetic route is as follows:

the preparation method of the compound (II) is as follows:

the method comprises the steps of using optical amino acid as a raw material, sequentially protecting carboxyl groups through esterification, carrying out condensation through triphosgene, connecting another optical amino acid with the carboxyl groups protected through esterification, removing protecting groups to be removed, and carrying out corresponding reaction on methyl ester to obtain a target compound.

The synthetic route is as follows:

the preparation method of the compound (III) is as follows:

the target compound is obtained by using benzyloxycarbonyl lysine as a raw material, sequentially protecting amino with tert-butoxycarbonyl, condensing with various alcohols, phenols and amines, removing tert-butoxycarbonyl, condensing with triphosgene to connect related groups, removing benzyloxycarbonyl, or continuing to condense with corresponding acyl chloride or anhydride, or adding other amino protecting groups.

The synthetic route is as follows:

r is as defined above₁、R₁’、R₂X, Y as defined above for peptoid compounds of general formula (I), (II) or (III);

reagents and reaction conditions in the above reaction formula: (a) acetyl chloride, anhydrous methanol, reflux, (b) triphosgene, saturated aqueous sodium bicarbonate, dichloromethane, 0 ℃, (c) as used, the various aforementioned amines, triethylamine, dichloromethane, (d) potassium hydroxylamine, anhydrous methanol, 25 ℃, (e) as used, triphosgene, toluene, reflux, (f) as used, the various aforementioned amino acid methyl ester hydrochlorides, triethylamine, dichloromethane, (g) as used, the various aforementioned amino acid methyl ester hydrochlorides, triethylamine, dichloromethane, (h) as used, p-toluenesulfonic acid, benzyl alcohol, 90 ℃, (i) as used, the various aforementioned amino acid methyl ester hydrochlorides, triethylamine, dichloromethane, (j) palladium/carbon, hydrogen, anhydrous methanol, 30 ℃, (k) as used, the various aforementioned amino acid methyl ester hydrochlorides, triethylamine, dichloromethane, (l) preparation of carboxylic acids: 1mol/L sodium hydroxide and methanol at 25 ℃; preparation of hydroxamic acid: potassium hydroxylamine, anhydrous methanol, 25 ℃, (m) (Boc)₂O, aqueous sodium hydroxide solution, tetrahydrofuran, room temperature, (n) as used, the various alcohols, phenols or amines mentioned above, EDCI, HOBt, dichloromethane, room temperature, (O) saturated solution of HCl in ethyl acetate, room temperature, (p) triethylamine, dichloromethane, room temperature, (q) palladium on carbon, hydrogen, anhydrous methanol, 30 ℃, (r) as used, the acid chlorides or acid anhydrides corresponding to the various acyl groups mentioned above.

The specific procedures for the compounds are described in detail in the examples.

The skilled person can vary the above steps to increase the yield, and can determine the synthetic route according to the basic knowledge in the art, such as choice of reactants, solvents and temperature, and can increase the yield by using various conventional protecting groups to avoid side reactions. These conventional protection methods can be found, for example, in T.Greene, Protecting Groups in organic Synthesis.

Obviously, the above-mentioned route is a stereoselective synthesis, and an optically active peptoid compound can be prepared by the above-mentioned route. For example, various amino acids as starting materials are replaced with optical isomers thereof (D configuration). Various other isomers of such ureido-like peptide derivatives are readily available to those skilled in the art and may be purified by conventional separation means, such as chiral salts or chiral chromatography columns, and the like.

The assay for aminopeptidase N inhibitory activity is described in Lejczak, B et al Biochemistry, 1989, 28, 3549. The substrate L-leucyl-p-nitroaniline is degraded by aminopeptidase N to produce p-nitroaniline having an absorption at 405nm, and the concentration of p-nitroaniline is positively correlated with the magnitude of the enzyme activity. The content of p-nitroaniline is determined by detecting the absorbance at 405nm, so that the activity of aminopeptidase is determined, and the degree of inhibition of the inhibitor on the enzyme activity is indirectly reflected.

The in vitro enzyme inhibition test of the peptide-like compound with the general formula (I), (II) or (III) proves that the peptide-like compound is a carbamido peptide aminopeptidase N inhibitor. The ureido peptoids of the invention are spatially matched to the active site of aminopeptidase N and thus exhibit high aprotinin activity in vitro.

Ubenimex, an aminopeptidase N inhibitor, has been on the market for many years in clinical use for the treatment of leukemia and can be used as an immunomodulator. The compound of the invention is used as the application of the carbamido peptide aminopeptidase N inhibitor. Has wide application prospect in preparing medicaments for preventing or treating mammal diseases related to the abnormal expression of the aminopeptidase N activity. According to the prior art, the mammalian diseases associated with the abnormal expression of aminopeptidase N activity include: inflammation, cancer, multiple sclerosis, various tissue ulcers or ulcerative conditions, periodontal disease, epidermolysis bullosa and leukemia.

Pharmaceutical compositions containing the compounds of the invention:

the partial derivatives of the invention may be present in free form or in salt form. Pharmaceutically acceptable salts of many compound types and methods for their preparation are known to those skilled in the art. Pharmaceutically acceptable salts include conventional non-toxic salts, including salts of such compounds formed from acids and inorganic or organic bases.

The compounds of the present invention may form hydrates or solvates. The person skilled in the art is aware of methods for forming hydrates when compounds are lyophilized together with water or solvates when concentrated in solution with suitable organic solvents.

The invention encompasses pharmaceutical compositions comprising a therapeutic amount of a compound of the invention, and one or more pharmaceutically acceptable carriers and/or excipients. Carriers include, for example, saline, buffered saline, dextrose, water, glycerol, ethanol, and combinations thereof, as discussed in more detail below. The composition may also contain minor amounts of wetting or emulsifying agents, or pH buffering agents, if desired. The composition can be in the form of a liquid, suspension, emulsion, tablet, pill, capsule, sustained release formulation or powder. The composition can be formulated into suppository with conventional binder and carrier such as triglyceride. Oral formulations may include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose and magnesium carbonate, and the like. Depending on the desired formulation, the formulation may be designed to mix, granulate and compress or dissolve the ingredients. In another approach, the composition may be formulated as nanoparticles.

The pharmaceutical carrier used may be, for example, a solid or a liquid.

Typical solid carriers include lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid and the like. The solid carrier may comprise one or more substances which may act simultaneously as flavouring agents, lubricants, solubilizers, suspending agents, fillers, glidants, compression aids, binders or tablet-disintegrants; it may also be an encapsulating material. In powders, the carrier is a finely divided solid which is in admixture with the finely divided active ingredient. In tablets the active ingredient is mixed with the carrier having the necessary compression properties in suitable proportions and compacted in the shape and size desired. Powders and tablets preferably contain up to 99% active ingredient. Suitable solid carriers include, for example, calcium phosphate, magnesium stearate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose, methyl cellulose, sodium carboxymethyl cellulose, polyvinylpyrrolidone, low melting waxes and ion exchange resins.

Typical liquid carriers include syrup, peanut oil, olive oil, water, and the like. Liquid carriers are used to prepare solutions, suspensions, emulsions, syrups, tinctures and sealed compositions. The active ingredient may be dissolved or suspended in a pharmaceutically acceptable liquid carrier such as water, an organic solvent, a mixture of both or a pharmaceutically acceptable oil or fat. The liquid carrier may contain other suitable pharmaceutical additives such as solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickening agents, pigments, viscosity regulators, stabilizers or osmo-regulators. Suitable examples of liquid carriers for oral and parenteral administration include water (containing in part additives as described above, e.g., cellulose derivatives, preferably sodium carboxymethyl cellulose solution), alcohols (including monohydric alcohols and polyhydric alcohols, e.g., glycols) and their derivatives, and oils (e.g., fractionated coconut oil and arachis oil). The carrier for parenteral administration may also be an oil or fat such as ethyl oleate and isopropyl myristate. Sterile liquid carriers are sterile liquid compositions for parenteral administration. The liquid carrier for the pressurized composition may be a halogenated hydrocarbon or other pharmaceutically acceptable propellant. Sterile solutions or suspension solutions liquid pharmaceutical compositions may be used, for example, for intravenous, intramuscular, intraperitoneal or subcutaneous injection. The injection can be performed by single push or gradual injection, and is infused into channels and collaterals for 30 minutes. The compounds may also be administered orally in the form of liquid or solid compositions.

The carrier or excipient may comprise a time delay material known in the art, such as glyceryl monostearate or glyceryl distearate, and may also comprise a wax, ethylcellulose, hydroxypropylmethylcellulose, methylmethacrylate, or the like. When the formulation is intended for oral administration, it is recognized that 0.01% tween 80 in phostapg-50 (phosphoipid and 1, 2-propanediol concentrate, a. nattermann & cie. gmbh) is used in the formulation of acceptable oral formulations of other compounds, and may be adapted to the formulation of various compounds of the invention.

A wide variety of pharmaceutical forms can be used in administering the compounds of the present invention. If a solid carrier is used, the preparation may be in the form of a tablet, powder or pellet placed in a hard gelatin capsule or in the form of a lozenge or troche. The amount of solid carrier varies widely, but is preferably from about 25mg to about 1.0 g. If a liquid carrier is used, the preparation may be in the form of a syrup, emulsion, soft gelatin capsule, sterile injectable solution or suspension in an ampoule or vial or in a nonaqueous liquid suspension.

To obtain a stable water-soluble dosage form, the compound or a pharmaceutically acceptable salt thereof may be dissolved in an aqueous solution of an organic or inorganic base. If a soluble form is not obtained, the compound may be dissolved in a suitable co-solvent or combination thereof. Examples of such suitable co-solvents include, but are not limited to, ethanol at a concentration ranging from 0 to 60% by volume of the total volume, propylene glycol, polyethylene glycol 300, polysorbate 80, glycerol, polyoxyethylene fatty acid esters, fatty alcohols or glycerol hydroxy fatty acid esters, and the like.

Various delivery systems are known and may be used for the administration of compounds or other various formulations including tablets, capsules, injectable solutions, capsules in liposomes, microparticles, microcapsules, and the like. Methods of introduction include, but are not limited to, cutaneous, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, pulmonary, epidural, ocular and (generally preferred) oral routes. The compounds may be administered by any convenient or other suitable route, for example by infusion or bolus injection, by absorption through epithelial or mucosal lines (e.g., oral mucosa, rectal and intestinal mucosa, etc.) or by drug-loaded stents and may be administered together with other biologically active agents. Can be administered systemically or locally. For use in the treatment or prevention of a hetero-, bronchial or pulmonary disorder, the preferred route of administration is oral, nasal or bronchial aerosol or nebulizer.

The design of the present invention involving compounds of the general formula (I), (II) or (III) as described above employs peptoid and isostere design strategies. Peptoid and isostere strategies have been widely applied to the field of design and development of antiviral and antitumor drugs, the structure of the peptoid and isostere strategies is similar to that of a peptide composed of natural or unnatural amino acids and non-peptide groups, but the overall conformation of the peptoid and isostere strategies is different from that of a natural polypeptide substance, on one hand, the peptoid has the intrinsic activity of a substrate, the activity of an enzyme can be inhibited by recognizing the active center of the enzyme, and meanwhile, the selectivity and the efficiency of a target site are improved; in addition, the peptoid and the natural peptide have structural difference, so that the peptoid is not easily degraded by peptidase, the biological stability and the utilization rate are improved, and the compound has long action time.

Specifically, the method takes optical purity amino acid as a raw material, selectively protects carboxyl, and synthesizes key intermediates through steps of substitution, condensation, deprotection and the like, and aims to enhance the affinity and metabolic stability of the compound with enzyme or receptor.

The aminopeptidase N inhibitor with a brand-new mother nucleus designed and synthesized by the invention has no cytotoxic activity but shows obvious in-vitro enzyme inhibitory activity as shown by in-vitro experiments, and is expected to become a non-cytotoxic anticancer candidate drug.

Detailed Description

The present invention will be further described with reference to the following examples, but is not limited thereto.

EXAMPLE 1 Synthesis of Compound (I) of the present invention

1. Taking (2S) -2-isopropyl-2- (3-benzylureido) -N-hydroxyacetamide (3b) as an example:

1) preparation of benzyl isocyanate (1)

A solution of 1.16g of benzylamine in 20mL of toluene was added dropwise to 80mL of a toluene solution of 2.22g of triphosgene at room temperature, and the reaction mixture was refluxed for 4 hours, and then toluene was evaporated to dryness to obtain benzyl isocyanate (1) as a yellow oily product, which was used in the next step without further purification.

2) Preparation of methyl 2- (3-benzylureido) -2-isopropylacetate (2)

After the dichloromethane solution of benzyl isocyanate (1) was dropped into the dichloromethane solution containing 2.51g L-valine methyl ester hydrochloride and 2.12g of triethylamine in ice bath, the reaction solution was stirred at room temperature, after 1 hour, dichloromethane was evaporated to dryness, the residue was dissolved in ethyl acetate, and then the mixture was washed with 1N hydrochloric acid and saturated brine, and the organic phase was dried over magnesium sulfate. Methyl 2- (3-benzylureido) -2-isopropylacetate (2) was obtained as a yellow oil which was used in the next step without further purification.

3)NH₂Preparation of OK

Dropping 140mL of saturated anhydrous methanol solution of KOH into 240mL of anhydrous methanol solution containing 46.7g (670mmol) of hydroxylamine hydrochloride, controlling the internal temperature to be lower than 40 ℃, cooling the reaction solution after dropping, filtering off white KCl precipitate, and hermetically storing the obtained filtrate for later use.

4) Preparation of (2S) -2-isopropyl-2- (3-benzylureido) -N-hydroxyacetamide (3b)

Methyl 2- (3-benzylureido) -2-isopropylacetate (2) was added to the above 30mL of hydroxylamine potassium in methanol, stirred at room temperature and monitored by TLC for progress. After completion of the reaction, methanol was distilled off under reduced pressure, the residue was dissolved in 1N hydrochloric acid, pH was adjusted to 1-2, ethyl acetate was added for extraction, the organic phase was washed with saturated brine and dried. Crude product (2S) -2-isopropyl-2- (3-benzylureido) -N-hydroxyacetamide (3b) was obtained as an off-white solid, which was separated by column chromatography (petroleum ether: ethyl acetate: 2: 1) to obtain 2.47g of (2S) -2-isopropyl-2- (3-benzylureido) -N-hydroxyacetamide (3b) as a white solid in 68% overall yield. mp 180-. ESI-MS m/z: 265.3[ M + H]⁺。1H-NMR(300MHz，DMSO-d6)：δ10.61(s，1H)，δ8.80(s，1H)，δ7.33-7.19(m，5H)，δ6.50(t，1H，J＝6.0Hz)，δ6.16(d，1H，J＝9.3Hz)，δ4.20(d，2H，J＝6.0Hz)，δ3.87(dd，1H，J＝7.2Hz，9.3Hz)，δ1.85-1.74(m，1H)，δ0.85(d，6H，6.9Hz)。

Example 2 Synthesis of Compound (III) of the present invention

Using (2S) -6-benzyloxycarbonyl-2- (3-methoxycarbonylmethyl-ureido) hexanoic acid methyl ester (1h) as an example:

1) preparation of N6-benzyloxycarbonyl-L-lysine methyl ester hydrochloride (3)

Slowly dropwise adding 50mL of anhydrous methanol into 2.35g of acetyl chloride in an ice bath, stirring at room temperature for half an hour after dropwise adding, then adding 2.80g N6-benzyloxycarbonyl-L-lysine once, then slowly heating the reaction mixture to reflux, and evaporating the methanol after 1 hour to obtain a white solid compound N6-benzyloxycarbonyl-L-lysine methyl ester hydrochloride (3), wherein the white solid compound can be directly used for the next reaction without further purification.

2) Preparation of N6-benzyloxycarbonyl-L-lysine methyl ester-1-isocyanate (4)

N6-benzyloxycarbonyl-L-lysine methyl ester hydrochloride (3) obtained in the above step 1) was added to 40mL of a saturated sodium bicarbonate solution and 40mL of dichloromethane, and 1.00g of triphosgene was added at a time with stirring in an ice bath. Stirring vigorously in ice bath for 15min, separating, extracting water layer with dichloromethane three times, combining organic layers, and drying. The solvent was evaporated to dryness to give compound N6-benzyloxycarbonyl-L-lysine methyl ester-1-isocyanate (4), which was used directly in the next reaction without purification.

3) Preparation of Glycine methyl ester hydrochloride (5)

Slowly dropwise adding 50mL of anhydrous methanol into 2.35g of acetyl chloride in an ice bath, stirring at room temperature for half an hour after dropwise adding, then adding 0.75g of glycine once, then slowly heating the reaction mixture to reflux, and after 1 hour, evaporating the methanol to dryness to obtain a white solid compound glycine methyl ester hydrochloride (5), wherein the white solid compound glycine methyl ester hydrochloride can be directly used for the next reaction without further purification.

4) Preparation of (2S) -6-benzyloxycarbonyl-2- (3-methoxycarbonylmethyl-ureido) hexanoic acid methyl ester (1h)

Glycine obtained in the above step 3)Acid methyl ester hydrochloride (5) and 1.01g triethylamine were stirred in 20mL dichloromethane, 10mLN 6-carbobenzoxy-L-lysine methyl ester-1-isocyanate (4) solution in dichloromethane was added dropwise in ice bath, after the addition was completed, stirring was performed at room temperature for one hour, dichloromethane was evaporated to dryness, the residue was dissolved in ethyl acetate, washed with 1N hydrochloric acid and saturated brine, and then the organic phase was dried. The solvent was evaporated to dryness to give crude (2S) -6-benzyloxycarbonyl-2- (3-methoxycarbonylmethyl-ureido) methyl hexanoate (1h) as a yellow oil, which was separated by column chromatography (petroleum ether: ethyl acetate: 1: 4) to give 1.48g of methyl (2S) -6-benzyloxycarbonyl-2- (3-methoxycarbonylmethyl-ureido) hexanoate as a white solid (1h), in 39% overall yield. ESI-MS m/z: 410.2[ M + H]⁺。1H-NMR(600MHz，CDCl₃)：δ7.36-7.26(m，5H)，δ5.13-5.04(m，3H)，δ4.48-4.44(m，2H)，δ3.73(s，3H)，δ3.71(s，3H)，δ3.21-3.17(m，2H)，δ1.81-1.35(m，6H).

EXAMPLE 3 Synthesis of Compound (II) of the present invention

Taking 2- [3- (1-hydroxycarbamoyl-2-phenyl-ethyl) -ureido ] -3-phenylpropionic acid hydroxyamide (4g) as an example:

1) preparation of phenylalanine methyl ester hydrochloride (6)

Slowly dropwise adding 100mL of anhydrous methanol into 4.24g of acetyl chloride in an ice bath, stirring at room temperature for half an hour after dropwise adding, then adding 2.98g of phenylalanine once, then slowly heating the reaction mixture to reflux, and evaporating the methanol after 1 hour to obtain a white solid compound, namely phenylalanine methyl ester hydrochloride (6), wherein the white solid compound can be directly used for the next reaction without further purification.

2) Preparation of phenylalanine methyl ester-1-isocyanate (7)

1.90g of phenylalanine methyl ester hydrochloride (6) obtained in the above step 1) was added to 40mL of a saturated sodium bicarbonate solution and 40mL of dichloromethane, and 1.00g of triphosgene was added at a time with stirring in an ice bath. Stirring vigorously in ice bath for 15min, separating, extracting water layer with dichloromethane three times, combining organic layers, and drying. The solvent is evaporated to dryness to obtain the compound phenylalanine methyl ester-1-isocyanate (7), which can be directly used for the next reaction without purification.

3) Preparation of methyl 2- [3-1- (methoxycarbonyl-2-phenyl-ethyl) -ureido ] -3-phenylpropionate (8)

Stirring 1.90g of phenylalanine methyl ester hydrochloride (6) obtained in the step 1) and 1.01g of triethylamine in 20mL of dichloromethane, dropwise adding a dichloromethane solution of phenylalanine methyl ester-1-isocyanate (7) in ice bath, stirring at room temperature for one hour after the dropwise adding is finished, evaporating dichloromethane, dissolving the residue in ethyl acetate, washing with 1N hydrochloric acid and saturated saline solution, and drying the organic phase. The solvent was evaporated to dryness to give a crude 2- [3-1- (methoxycarbonyl-2-phenyl-ethyl) -ureido ] -3-phenylpropionic acid methyl ester (8) as a yellow oil, which was used directly in the next reaction without further purification.

4) Preparation of 2- [3- (1-hydroxycarbamoyl-2-phenyl-ethyl) -ureido ] -3-phenylpropionic acid hydroxyamide (4g)

Reacting 2- [3-1- (methoxycarbonyl-2-phenyl-ethyl) -ureido]Methyl-3-phenylpropionate (8) was added to the above 30mL of hydroxylamine potassium in methanol, stirred at room temperature and monitored by TLC for progress. After the reaction was completed, methanol was distilled off under reduced pressure, the residue was dissolved in 1N hydrochloric acid, pH was adjusted to 1-2, a white precipitate was precipitated, and the precipitate was washed with ethyl acetate and then dried in vacuum. To give the product 2- [3- (1-hydroxycarbamoyl-2-phenyl-ethyl) -ureido]-3-phenylpropionic acid hydroxyamide (4g) was 1.89g of white solid, 54% overall yield. ESI-MS m/z: 386.2[ M + H]⁺。1H-NMR(300MHz，DMSO-d6)：δ10.71(s，1H)，δ10.56(s，1H)，δ8.93(s，1H)，δ8.81(s，1H)，δ7.32-7.07(m，10H)，δ6.38-6.35(m，2H)，δ4.05-4.18(m，2H)，δ2.84-2.51(m，4H).

EXAMPLE 4 Activity test of the object Compound for inhibition of aminopeptidase N (In vitro)

The test principle and the detailed test steps are referred to CN 1974554A cyclic imide peptide metalloprotease inhibitor and the application thereof, and the test results are shown in Table 1.

TABLE 1 results of in vitro enzyme inhibition assay

^aNA＝not available

The test data in the table show that the inhibition activity of the compounds 4b, 5b, 6b, 7b, 4c, 6c, 4d and 4e on the aminopeptidase N is better than that of a positive control drug ubenimex, and the compound has good development prospect.

Claims

1. A compound of general formula (I):

wherein,

R₁is hydrogen, a substituent on the alpha carbon of various natural or unnatural amino acids: alanine, valine, phenylalanine, isoleucine, leucine, methionine, lysine, ornithine, aspartic acidAsparagine, glutamic acid, glutamine, serine, threonine, histidine, tryptophan, arginine, cysteine, citrulline, proline, hydroxyproline, tyrosine; beta-aminopropionic acid, beta-aminophenylpropionic acid; gamma-aminobutyric acid;

R₂is hydrogen, C1-12 fatty alkanyl, optionally substituted with one or more of the following: hydroxyl, halogen, nitro, cyano, guanidino, carboxyl.

2. A compound characterized by being one of the following:

1 a: 2- (3-tert-butylureido) -N-hydroxyacetamide,

2 a: (2S) -2-methyl-2- (3-tert-butylureido) -N-hydroxyacetamide,

3 a: (2S) -2-isopropyl-2- (3-tert-butylureido) -N-hydroxyacetamide,

4 a: (2S) -2- (2-methyl) -propyl-2- (3-tert-butylureido) -N-hydroxyacetamide,

5 a: (2S) -2- (1-methyl) -propyl-2- (3-tert-butylureido) -N-hydroxyacetamide,

6 a: (2S) -2-benzyl-2- (3-tert-butylureido) -N-hydroxyacetamide,

7 a: (2S) -2- (methylthio) -ethyl-2- (3-tert-butylureido) -N-hydroxyacetamide,

1 b: 2- (3-benzylureido) -N-hydroxyacetamide,

2 b: (2S) -2-methyl-2- (3-benzylureido) -N-hydroxyacetamide,

3 b: (2S) -2-isopropyl-2- (3-benzylureido) -N-hydroxyacetamide,

4 b: (2S) -2- (2-methyl) -propyl-2- (3-benzylureido) -N-hydroxyacetamide,

5 b: (2S) -2- (1-methyl) -propyl-2- (3-benzylureido) -N-hydroxyacetamide,

6 b: (2S) -2-benzyl-2- (3-benzylureido) -N-hydroxyacetamide,

1 c: 2- (3-phenethylureido) -N-hydroxyacetamide,

2 c: (2S) -2-methyl-2- (3-phenethylureido) -N-hydroxyacetamide,

3 c: (2S) -2-isopropyl-2- (3-phenethylureido) -N-hydroxyacetamide,

4 c: (2S) -2- (2-methyl) -propyl-2- (3-phenethylureido) -N-hydroxyacetamide,

5 c: (2S) -2- (1-methyl) -propyl-2- (3-phenethylureido) -N-hydroxyacetamide,

6 c: (2S) -2-benzyl-2- (3-phenethylureido) -N-hydroxyacetamide,

7 c: (2S) -2- (methylthio) -ethyl-2- (3-phenethylureido) -N-hydroxyacetamide or

3. A process for the preparation of a compound according to claim 1, characterized in that:

optical amino acid is taken as a raw material, carboxyl is protected by methyl esterification in sequence, corresponding amine is connected by triphosgene through condensation, and then the methyl ester is subjected to corresponding reaction to obtain a target compound; or corresponding amine is taken as a raw material, is converted into isocyanate by triphosgene in sequence, is condensed with methyl ester of optical amino acid, and then the methyl ester is subjected to corresponding reaction to obtain a target compound;

the synthetic route is as follows:

wherein R is₁、R₂A peptoid compound of general formula (i) as defined in claim 1;

reagents in the above reaction scheme: (a) acetyl chloride, anhydrous methanol, reflux (b) triphosgene, saturated aqueous sodium bicarbonate, dichloromethane, various of the above amines used at 0 ℃ (c), triethylamine, dichloromethane (d) potassium hydroxylamine, anhydrous methanol, 25 ℃ (e) triphosgene, toluene, and various of the above amino acid methyl ester hydrochlorides used at reflux (f), triethylamine, dichloromethane.

4. A pharmaceutical composition comprising a compound of claim 1 or 2 and one or more pharmaceutically acceptable carriers or excipients.