Aminopeptidase N inhibitor, preparation method and application

A technology for aminopeptidase and inhibitors, applied in the field of aminopeptidase N inhibitors and preparation, capable of solving problems such as limited sources and high cost of total synthesis

Inactive Publication Date: 2012-03-21
WEIFANG BOCHUANG INT ACAD OF BIOTECH & MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Ubenimex, as a dipeptide-like structure containing β-amino acids, is currently used as an immune enhancer for the treatment of leukemia. It is derived from Streptomyces olivorecticuli Isolated from the culture medium, the total synthesis is expensive and the source is limited

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  • Aminopeptidase N inhibitor, preparation method and application
  • Aminopeptidase N inhibitor, preparation method and application
  • Aminopeptidase N inhibitor, preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Aminopeptidase N inhibitor, chemical name: ( S )-4-Methyl-2-(3-naphthyl-1-ylmethyl-ureido)-pentanoylhydroxylamine. The structural formula of compound (I) is as follows:

[0029]

[0030] The preparation method, reaction steps and reaction formula of the compound are as follows:

[0031] It is the condensation of naphthyl methyl isocyanate and L-leucine methyl ester to form the corresponding urea-based intermediate, and the methyl ester is partially converted into hydroxamic acid to obtain compound (I). The reaction formula is as follows: Synthetic route:

[0032]

[0033] The reagents in the above synthetic route are: (a) triphosgene, toluene, reflux, 3 hours; (b) leucine methyl ester hydrochloride, triethylamine, dichloromethane, room temperature, one hour; (c) hydrochloric acid Hydroxylamine, potassium hydroxide, methanol, one hour.

[0034] Specific synthesis method:

[0035] 1) Naphthyl methyl isocyanate (1):

[0036] To the 1,4-dioxane solution of 1-naphthylmethylamine ...

Embodiment 2

[0042] Activity test of target compound to inhibit aminopeptidase N (In vitro)

[0043] 1. Principle: Aminopeptidase N interacts with its substrate (L-leucyl-p-nitroaniline) to produce p-nitroaniline with absorption at 405nm, and the concentration of p-nitroaniline and enzyme activity The size is positively correlated. Determine the content of p-nitroaniline by detecting the absorbance at 405nm, thereby determining the activity of aminopeptidase, which indirectly reflects the degree of inhibition of the enzyme activity by the inhibitor.

[0044] 2. Materials and methods:

[0045] Aminopeptidase N and the substrate L-leucyl-p-nitroaniline were purchased from Sigma

[0046] Solution preparation:

[0047] Buffer solution, prepare pH 7.2, 50mM phosphate buffer solution, and place at room temperature for later use.

[0048] Aminopeptidase N was dissolved in a buffer to make a 0.2mg / mL solution;

[0049] The substrate was dissolved in DMSO to prepare a 0.5 mg / mL solution, and each solution...

Embodiment 3

[0067] In vitro inhibitory activity of target compounds against cell invasion

[0068] Compound (I) was tested for the activity of inhibiting tumor cell invasion in vitro, and the results are shown in figure 1 .

[0069] Term description:

[0070] ES-2 is a human ovarian clear cell cancer cell line.

[0071] Bestatin, trade name Ubenmix (Ubenmix), is currently on the market as an immune enhancer for the treatment of leukemia. It is derived from Streptomyces olive ( Streptomyces olivorecticuli ) Small molecule peptide APN inhibitor isolated from the culture medium.

[0072] DMSO: Dimethyl sulfoxide.

[0073] Matrigel: Intercellular matrix extracted from mouse EHS sarcoma

[0074] 1. [Material] ES-2 cell line, pre-coated with Matrigel's Transwell chamber, crystal violet, RPMI-1640 medium containing 1% fetal bovine serum, RPMI-1640 medium containing 10% fetal bovine serum, 24 wells board

[0075] 2. [Method]

[0076] Cell culture The ES-2 cell line was cultured routinely. The logarithm...

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Abstract

The invention belongs to the technical field of drugs, and particularly relates to aminopeptidase N inhibitor, a preparation method and application to tumor resistance. The technical scheme includes that the aminopeptidase N inhibitor is chemically named as (S)-4-methyl-2-(3-naphthyl-1-ylmethyl-ureido)-valeryl group hydroxylamine. The constitutional formula (I) of the aminopeptidase N inhibitor is shown here. The pharmacodynamic effect of the aminopeptidase N inhibitor is evaluated by in-vitro APN (adiponectin) inhibitory experiments, in-vitro resistance to tumor cell invasion, antiangiogenesis and mouse in-vivo resistance to melanoma manual transfer and antiangiogenesis experiments.

Description

technical field [0001] The invention belongs to the technical field of medicines, and in particular relates to an aminopeptidase N inhibitor, a preparation method and an antitumor application. Background technique [0002] Aminopeptidase N (APN, CD13) is a type II membrane-bound glycoprotein with a molecular weight of about 150 kDa. It belongs to zinc ion-dependent metalloproteases, exists in the form of membrane-bound homodimers, and participates in various biological functional peptides Degradation of N-terminal neutral amino acids. Aminopeptidase N is widely distributed in various types of cells in the human body, including stromal cells of hematopoietic organs, endothelial cells of new blood vessels, and various epithelial cells including renal proximal tubule epithelial cells, bile duct epithelial cells and small intestinal brush border epithelial cells, etc. ; It is also distributed on the surface of fibroblasts, leukemia cells, mesenchymal tumor cells, ovarian cancer...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C275/24C07C273/18A61K31/17A61P35/00A61P29/00A61P37/06A61P35/02A61P1/02
Inventor 徐文方宿莉
Owner WEIFANG BOCHUANG INT ACAD OF BIOTECH & MEDICINE
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