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Polypeptide and use thereof in medicament preparation

一种一多肽、作用的技术，应用在生物制药及糖尿病领域，能够解决使用和功效限制等问题

Active Publication Date: 2010-12-15
刘建宁
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, drugs that can really treat type 2 diabetes have not yet been found, because the above drugs have their own limitations in use and efficacy

Method used

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  • Polypeptide and use thereof in medicament preparation
  • Polypeptide and use thereof in medicament preparation
  • Polypeptide and use thereof in medicament preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1: Nesfatin-1 mediates the effect of plasminogen in obese and diabetic animals

[0073] Nesfatin-1, a polypeptide fragment hypothesized to be derived from NUCB2, was recently identified as an anorexigenic factor associated with melanocortin signaling in the hypothalamus. Injection of nesfatin-1 intracerebroventricularly or intraperitoneally reduced food intake and consequently reduced body weight. The amino acid sequence of nesfatin-1 is highly conserved from mouse to human, and has several recognized enzyme cleavage sites for plasmin ( figure 1 a). Therefore, nesfatin-1 is hypothesized to mediate the effects of plasminogen in obese and diabetic animals.

[0074] To prove this hypothesis, we expressed and purified recombinant nesfatin-1 using genetically engineered Escherichia coli. It was co-incubated with plasmin, and as expected, nesfatin-1 was rapidly degraded ( figure 1 b). Interestingly, compared to the genotype for plg - / - lepr - / - Non-obese litt...

Embodiment 2

[0083] Example 2: Effect of Tail Vein Injection of Plasmin Inhibitors on the Concentration of Nesfatin-1 in Blood

[0084]The manner in which Nesfatin-1 is cleared from the peripheral blood is unclear. Trace amounts of plasmin production in peripheral blood were previously reported and confirmed in our experiments. Inject AMCA and aprotinin, two inhibitors of plasmin, into db / db mice through the tail vein, while the concentration of nesfatin-1 in the peripheral blood increases, the mice eat less and their body weight decreases ( Figure 5 ). In addition, nesfatin-1 injected by tail vein - / - The peripheral blood clearance rate of genotype mice is significantly slower than that of plg + / + of mice. Therefore, we believe that nesfatin-1 in peripheral blood is at least partially degraded by plasmin. Consistent with reports that nesfatin-1 can penetrate the blood-brain barrier, AMCA can also cause anorexia, indicating that nesfatin-1 in the brain is at least partially derived f...

Embodiment 3

[0086] Example 3: Quantitative PCR detection of neuropeptides

[0087] For the detection of neuropeptide mRNA quantitative PCR (q-PCR), the CFX96TM real-time system (Bio-Rad, Hercules, CA) and the detection method of SYBR Green I fluorescent dye were used. Briefly, hypothalamus tissue from 24-hour fasted mice was homogenized, and total RNA was extracted using RNAiso reagent (TaKaRa, Dalian, CN) and reverse transcribed into single-stranded cDNA. We determined the relative expression of neuropeptide mRNA using a standard curve of hypothalamic cDNA and adjusted for total RNA and gadph RNA by qPCR. The primers used in real-time RT-PCR are as follows: agrp forward primer: 5'-TGT GTA AGG CTG CAC GAG TC (SEQ ID NO: 10); agrp reverse primer: 5'-GGC AGTAGC AAA AGG CAT TG (SEQ ID NO : 11); agrp Tm: 61°C; npy forward primer: 5′-AGG CTTGAA GAC CCT TCC AT (SEQ ID NO: 12); npy reverse primer: 5′-ACA GGC AGA CTG GTTTCA GG (SEQ ID NO : 13); npy Tm: 61°C; pomc forward primer: 5'-CGC CCG TGT ...

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Abstract

The present invention belongs to the field of bio-pharmaceuticals and diabetes, and discloses a polypetide and use of the polypetide in medicament preparation. The polypetide is (a) or (b): (a) polypeptide which is composed of amino acid sequence represented by SEQ ID NO:22; (b) polypetide which is composed of the amino acid sequence in (a) through replacement, deletion or adding one or several amino acids, has blood sugar reduction and / or blood fat reduction function and is derived from (a). The polypeptide and a plasmin inhibitor thereof have the following functions: reducing blood sugar, reducing triglyceride, total cholesterol or LDL in blood. The polypeptide can be used for preparing the blood sugar reduction medicament or blood fat reduction medicament, and particularly for the diabetes treatment medicament.

Description

technical field [0001] The invention belongs to the fields of biological pharmacy and diabetes, and relates to a polypeptide (peptide segment of NUCB2, such as pladin or nesfatin-1) and its application in pharmacy. Background technique [0002] The incidence of diabetes is increasing at an alarming rate worldwide, especially in developing countries. Excessive blood sugar in diabetics can damage blood vessels, nerves, eyes, and kidneys, eventually leading to severe cardiovascular disease, neurosis, blindness, and kidney failure. Therefore, how to control blood sugar is the most critical issue to be faced in the treatment of diabetes. [0003] In people with type 1 diabetes who are insulin deficient, injecting insulin before food intake can prevent hyperglycemia. However, for patients with decreased insulin expression or insulin resistance, namely type II diabetics, rising blood sugar level is one of many complicated factors that cause morbidity and death. Many drugs are no...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C07K7/08A61K38/17A61K38/10A61P3/10A61P3/06
CPCG01N2800/042A61K38/1709A01K2267/0362G01N33/5088A61K38/57A61P3/06A61P3/10
Inventor 刘建宁
Owner 刘建宁
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