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Polypeptide, composition and application thereof

A composition and drug technology, applied in the field of biopharmaceuticals, can solve problems such as inability to exert a good therapeutic effect and the complexity of sepsis

Active Publication Date: 2021-09-14
北京鲲达宇科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the clinical treatment of sepsis mainly relies on antibiotics and other adjuvant treatment measures, but these methods often cannot play a good therapeutic role due to drug resistance, antibiotic toxicity and the complexity of sepsis.

Method used

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  • Polypeptide, composition and application thereof
  • Polypeptide, composition and application thereof
  • Polypeptide, composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0100] Example 1 Gradual separation of active ingredients of collagen 1

[0101] (1) First, dissolve donkey collagen 1 (Advanced Biomatrix company product number: 5005) in deionized water at 50 °C to obtain a collagen solution, cool it to 37 °C, and adjust the pH of the collagen solution to 2.0 (It is recommended to use a pH agent for accurate 1 N dilute hydrochloric acid for pH adjustment), add pepsin (Sigma company, product number EC3.4.23.1) according to the weight ratio of pepsin and collagen solution 1:250, at 37 ℃ , digested for 2 hours; then the pH value of the resulting mixture was adjusted to 6.8, and trypsin (Sigma company, product number EC3.4.21.4) was added at a ratio of 1:250 by weight of trypsin and collagen solution. Also at 37 Digest at ℃ for 2h. Subsequently, the collagen solution digested with pepsin and trypsin was heated to 100 °C for 1 hour to inactivate the enzymes. After the solution was naturally cooled, centrifugation was performed at 8000 rpm for 10...

Embodiment 2

[0109] Example 2 The anti-cell death A component is separated by ion exchange chromatography

[0110] (1) The A component (molecular weight ≥ 10,000 Da) is separated step by step by ion exchange chromatography. Ion exchange chromatography (IEX) uses IEX technology to purify biomolecules and separate them according to the difference in their net surface charges. This application used AKTA-pure (GE Healthcare, USA) equipped with High-Performance Q (GE healthcare, USA) anion exchange column (100mm*10mm Sepax, USA). Elution was performed with a constant gradient of 50% 1 M NaCl, 50 Mm Tris pH 7.4 at a flow rate of 3 ml / min at room temperature. Absorbance was monitored at 214 and 280 nm. Fractions at the peaks were collected at both A1 and A2 permeabilized and then lyophilized. The lyophilized solution was dissolved in ultrapure water for analysis. For the analysis results, see figure 2 A.

[0111] (2) Detection of the protective effects of the two components A1 and A2 on hyd...

Embodiment 3

[0113] Example 3A1 components were further separated by reversed-phase chromatography

[0114] (1) The A1 component is separated step by step by reversed-phase chromatography, and the RPC separates the fractions according to the hydrophobicity of the substance. The present application used AKTA-pure (GE healthcare, USA) equipped with Source30-RPC (GE healthcare, USA) reversed-phase chromatography column (65mm*10mm Sepax) for fractionation. That is, gradient elution was performed with 0-10% linner first, followed by constant gradient elution with 10 mM Na2HPO4 (pH 7.0) buffer containing 20%, 30%, 50% and 60% acetonitrile, and the flow rate at room temperature was 3ml / min. Finally measure the absorbance at 214 and 280 nm. Peak fractions were collected and lyophilized. The lyophilized solution was dissolved in ultrapure water for analysis. For the analysis results, see image 3 A.

[0115] (2) Detection of the protective effects of four components A11, A12, A13 and A14 on ...

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PUM

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Abstract

The invention relates to a polypeptide, a composition and application thereof. The polypeptide is as shown in (i) or (ii) that (i), an amino acid sequence is as shown in SEQ ID NO: 1; and (ii) a polypeptide which is obtained by substitution and / or deletion and / or addition of one or more amino acid residues on the amino acid sequence as shown in SEQ ID NO: 1 and has the same function as the amino acid sequence as shown in SEQ ID NO: 1. The polypeptide has the effect of resisting cell death.

Description

technical field [0001] This application generally relates to the field of biopharmaceuticals, and specifically relates to the development and application of small molecule polypeptides, including the dosage form, dosage, and therapeutic effects of diseases of the polypeptides. Background technique [0002] Polypeptides are a class of compounds formed by connecting multiple amino acids through peptide bonds, usually composed of 10-100 amino acid molecules, the connection method is the same as that of proteins, and the relative molecular mass is less than 10,000. In recent years, with the development and maturity of peptide synthesis technology, peptide drugs have become one of the hot spots in drug research and development. They have been widely used in tumors, cardiovascular and cerebrovascular diseases, hepatitis, Prevention, diagnosis and treatment of diseases such as diabetes and AIDS. And these polypeptides play key roles in human physiology, which can act as hormones, ...

Claims

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Application Information

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IPC IPC(8): C07K14/78A61K47/64A61K49/00A61P3/10A61P7/00A61P9/10A61P25/14A61P25/16A61P25/28A61P31/04A61P35/00A61P37/02
CPCC07K14/78A61K47/6435A61K49/0056A61P31/04A61P7/00A61P9/10A61P25/28A61P25/16A61P25/14A61P37/02A61P35/00A61P3/10
Inventor 韩晶杨泽刘磊
Owner 北京鲲达宇科技有限公司
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