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A kind of erythropoietin mimetic peptide and its dimer, as well as preparation method and application

A technology of erythropoietin and peptide mimics, which is applied in the fields of erythropoietin, extracellular fluid diseases, chemical instruments and methods, etc., can solve the problems of low and limited EC50, and achieve the effect of prolonging the half-life

Active Publication Date: 2019-01-08
TIANJIN INSTITUTE OF PHARMA RESEARCH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the EC50 of peptides that can stimulate erythrocyte proliferation and differentiation is very low, between 20nM and 250nM, so the clinical application of these peptides is greatly limited

Method used

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  • A kind of erythropoietin mimetic peptide and its dimer, as well as preparation method and application
  • A kind of erythropoietin mimetic peptide and its dimer, as well as preparation method and application
  • A kind of erythropoietin mimetic peptide and its dimer, as well as preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Synthesis of Erythropoietin Mimetic Peptide Dimer

[0043] The erythropoietin mimetic peptide of the present invention is prepared by the Fmoc solid-phase peptide synthesis method, and the CS 336X instrument produced by CSBio Company is used to synthesize the peptide of the present invention. The method of synthesis was carried out according to the manufacturer's instruction manual. The Fmoc solid-phase polypeptide synthesis method described herein refers to a synthetic method in which a polymer resin is used as a solid-phase reaction matrix to sequentially condense amino-terminal Fmoc-protected amino acids in the presence of a coupling reagent to synthesize a polypeptide. For its specific method, see Fmoc solid phase peptide synthesis: a practical approach, 2000, Oxford University Press. And the disulfide bonds in the monomer are formed by oxidation methods, such as 20% DMSO oxidation method and iodine oxidation method. The prepared polypeptide was purifie...

Embodiment 2

[0047] Example 2 Effect of erythropoietin mimetic peptide dimer on mice

[0048] Using mice to evaluate and compare the effects of erythropoietin mimetic peptide and erythropoietin protein on mouse erythropoiesis.

[0049] Among them, EPO drugs were purchased from Shenyang Sansheng Pharmaceutical Co., Ltd.;

[0050] Kunming mice, purchased from Shanghai Experimental Animal Center of Chinese Academy of Sciences, weighing 25-30 g, are all female mice. The number of animals in each group in the experiment: 10, divided into 3 groups.

[0051] Among them, the mice in the experimental group were injected with erythropoietin mimic peptide dimer, the mice in the positive control group were injected with erythropoietin protein, and the mice in the blank control group were injected with PBS buffer at a dose of 4.5 mg / kg for three consecutive days. After 1 day, the mice were sacrificed, and the whole blood was taken to count the peripheral blood cells and reticulocytes, and the blood ...

Embodiment 3

[0055] Example 3: Effect of erythropoietin mimetic peptide dimer on macaques

[0056] The EPO drug used in the present invention was purchased from Shenyang Sansheng Pharmaceutical Co., Ltd.

[0057] Rhesus monkeys were used to evaluate the effect of erythropoietin-mimetic peptide dimers on erythropoiesis. Rhesus monkeys, weighing 5.5-8.5 kg, male or female, were purchased from Hainan Experimental Animal Center. The macaques were divided into two groups according to the basic hemoglobin, with three monkeys in each group. The experimental group used the mimetic peptide dimer of the present invention, intravenously injected once a week, 4.5 mg / kg each time; the other group was the positive control group used EPO, three times / week, 240 μg / kg each time, for five consecutive weeks , Measure hematological indicators once a week.

[0058] The results are shown in Table 3. A single intravenous injection of the mimetic peptide led to an increase in the hemoglobin content (32%) and ...

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Abstract

The invention relates to erythropoietin mimic peptide. The erythropoietin mimic peptide is characterized in that the amino acid sequence of monomer peptide of the mimic peptide is shown as GX1LYACHMGPITX2VCQPLRX3K in SEQ ID NO:1, wherein X1 is allylglycine (D-Allyiglycin), X2 is 3-(1-naphthyl)-L-alanine (Nal), X3 is sarcosine (Sar), 6-bit and 15-bit cysteine (C) forms an intramolecular disulfide bond, and N is acetylized in end. The invention further provides a preparation method of the mimic peptide, a preparation method of dimer and pharmaceutical salt of the mimic peptide. The invention further provides a pharmaceutical composition containing the mimic peptide or the dimer or the pharmaceutical salt of the mimic peptide. The erythropoietin mimic peptide, the dimer and the pharmaceutical salt of the mimic peptide can excite generation of erythrocyte and significantly prolong the half-life period of drugs in human bodies.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to an erythropoietin mimic peptide. An agonistic erythropoietin-mimicking peptide dimer and a preparation method thereof, and the present invention also relates to the use of the mimetic peptide dimer in the preparation of drugs for the treatment of diseases characterized by the lack of erythropoietin or the absence or deficiency of red blood cell populations use. Background technique [0002] Erythropoietin (EPO) is a glycoprotein hormone with a molecular weight of about 34kD. The erythropoietin present in the plasma is composed of 165 amino acids, with a high degree of glycosylation, and the main sugar component is sialic acid. According to different carbohydrate content, naturally occurring erythropoietin is divided into two types, α-type and β-type, among which, α-type contains 34% carbohydrates, and β-type contains 26% carbohydrates. The two types are identical in biological charact...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/505A61K38/18A61P13/12A61P7/06A61P37/02A61P35/00
CPCA61K38/00C07K14/505
Inventor 龚珉赵娜夏郑学敏王士伟魏群超夏广萍韩英梅周植星
Owner TIANJIN INSTITUTE OF PHARMA RESEARCH
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