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Erythropoietin mimic peptide and preparation method and application thereof

A technology of erythropoietin and mimetic peptides, which is applied in the field of erythropoietin mimetic peptides and its preparation and application, can solve the problems of low EC50 and limitation, and achieve the effect of prolonging the half-life

Active Publication Date: 2015-11-25
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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  • Erythropoietin mimic peptide and preparation method and application thereof
  • Erythropoietin mimic peptide and preparation method and application thereof
  • Erythropoietin mimic peptide and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Synthesis of Erythropoietin Mimetic Peptide

[0042] The present invention will be described in further detail below in conjunction with specific examples.

[0043] The erythropoietin mimetic peptide of the present invention is a polypeptide, which can be prepared using the Fmoc solid-phase polypeptide synthesis method, and the CS336X instrument produced by CSBio Company to synthesize the polypeptide 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 the specific method, see Fmocsolid phase peptide synthesis: practical approach, 2000, Oxford University Press. And form a cyclic polypeptide by amide...

Embodiment 2

[0044] Example 2 Effects of erythropoietin mimetic peptide on mice

[0045] Using mice to evaluate and compare the effects of erythropoietin mimetic peptide and erythropoietin protein on mouse erythropoiesis.

[0046] Among them, EPO drug (recombinant human erythropoietin injection, batch number: 201405YC12) was purchased from Shenyang Sansheng Pharmaceutical Co., Ltd.;

[0047] Kunming mice were purchased from the Shanghai Experimental Animal Center of the Chinese Academy of Sciences, weighing 25-30 g, all male mice, and the number of animals in each group in the experiment: 10, divided into 6 groups.

[0048] Wherein, the mice in the EPO group were injected with erythropoietin protein (EPO), the mice in the blank group were blank controls, injected with PBS buffer solution (25mM), and the mice in the experimental group were injected subcutaneously with the erythropoietin mimetic peptide SEQ ID NO: 2~SEQ ID NO :5, dissolved with PBS damping fluid (25mM), dosage is 4.5mg / kg...

Embodiment 3

[0052] Example 3 Effects of erythropoietin mimetic peptide on macaques

[0053] Rhesus monkeys were used to evaluate the effect of erythropoietin-mimetic peptides 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 5 groups according to the basic hemoglobin, with three monkeys in each group. The experimental group used the mimetic peptide of the present invention whose amino acid sequence is SEQIDNO: 2-5, intravenously injected once a week, 4.5mg / kg each time, and the control group used EPO, three times / week, 240μ / kg each time, continuous administration For five weeks, hematological indicators were measured once a week

[0054] The results are shown in Table 3. A single intravenous injection of the mimetic peptide led to an increase in the hemoglobin content (33%) and hematocrit in the peripheral blood of macaques, indicating that the mimetic peptide stimulated hemoglob...

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Abstract

The invention relates to erythropoietin mimic peptide capable of lastingly promoting erythropoiesis. Amino acid sequence of the erythropoietin mimic peptide is shown as SEQ ID NO:1GGLYAX1HMGPITX2VX3QPLRX4K, wherein X1 is chosen from lysine or arginine, X2 is 3-(1-naphthyl)-L-alanine, X3 is chosen from aspartic acid or glutamine, X4 is sarcosine, and free amino and carboxyl of an amino acid side chain are cyclically condensed between X1 and X3, and N terminal is acetylized. The invention further provides application of the erythropoietin mimic peptide in drug used for treating diseases featured by erythrocyte deficiency or erythrogenin deficiency or erythrocyte group deficiency or defect. The erythropoietin mimic peptide can promote generation of erythrocytes, and half-life period of the drug in vivo is prolonged remarkably.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a structurally similar erythropoietin mimic peptide modified by fatty acid that can bind to and activate erythropoietin receptors or can act as an agonist for erythropoietin The present invention also relates to the application of the structural analogue in the medicine for the treatment of diseases characterized by the lack of erythropoietin or the absence or defect of red blood cell population. 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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/505C07K1/04A61K38/18A61P13/12A61P7/08A61P7/06A61P37/02A61P35/00A61P29/00A61P39/06
CPCA61K38/1816C07K14/505A61K2300/00
Inventor 龚珉周植星王士伟郑学敏魏群超夏广萍赵娜夏韩英梅
Owner TIANJIN INSTITUTE OF PHARMA RESEARCH
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