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Peptide KAI 11 for promoting cartilage regeneration, and application thereof

A technology for cartilage and cartilage repair, applied in the field of polypeptides, to achieve broad application prospects, promote cartilage regeneration, and low immunogenicity

Active Publication Date: 2020-03-24
LINK HEALTH GRP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has not been reported in the literature that a certain polypeptide of BMP-7 has the activity of promoting cartilage regeneration

Method used

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  • Peptide KAI 11 for promoting cartilage regeneration, and application thereof
  • Peptide KAI 11 for promoting cartilage regeneration, and application thereof
  • Peptide KAI 11 for promoting cartilage regeneration, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The solid-phase synthesis of embodiment 1 peptide KAI 11

[0032] Peptide KAI 11 (SEQ ID NO.1) was synthesized by a conventional solid-phase process, and the purity of the synthesized peptide was >95% (see figure 1 ), synthesized 500mg.

[0033] Example 2 The effect of peptide KAI 11 on the differentiation of mesenchymal stem cells in vitro

[0034] 1) Cultivation and passage of human mesenchymal stem cells: resuscitate human bone marrow mesenchymal stem cells (BMSCs, purchased from Guangzhou Saiye Biotechnology Co., Ltd.), add complete medium, and place at 37°C, 5% CO 2 cultured in a humidified incubator. On the second day after recovery, the recovered cells were replaced with fresh complete medium, and the cells were replaced with fresh complete medium every two days until the cells reached 80%-90% confluence, and then subcultured.

[0035] 2) Chondrogenic induction: Peptide KAI 11 was dissolved in phosphate buffered solution (PBS). Cells were resuspended in compl...

Embodiment 3

[0043] Example 3 Effect of Peptide KAI 11 on Chondrocyte Proliferation

[0044] 1) Chondrocyte culture: Under aseptic conditions, the articular cartilage of 2-month-old New Zealand white rabbits was cut into 1mm size, digested with 2mg / ml hyaluronidase for 45 minutes, 2mg / ml trypsin for 45 minutes, and 37°C respectively. 4 mg / ml type II collagenase was digested for 3 hours, washed and centrifuged (1500 r / min) for 5 minutes, and the precipitate was placed in DMEM medium containing 15% fetal bovine serum for cultivation.

[0045] 2) Identification of chondrocytes: A small amount of primary chondrocytes were taken for smear and immunodetected with type I collagen antibody SABC, stained brownish yellow, indicating that type I collagen was secreted, and they were confirmed as chondrocytes.

[0046] 3) The proliferation of chondrocytes was measured by MTS method: chondrocytes were divided into 2×10 4 Each well was inoculated in a 96-well plate, and different concentrations of pepti...

Embodiment 4

[0054] Example 4 The repairing effect of peptide KAI 11 on the zebrafish cartilage injury model

[0055] Randomly select transgenic cartilage fluorescent zebrafish 2 days after fertilization in a six-well plate, 30 zebrafish per well (ie, each concentration group), and give dexamethasone in water to establish a zebrafish cartilage injury model. The model zebrafish were intravenously injected with peptide KAI 11 dissolved in normal saline at doses of 20, 100, and 500 ng / tail, and the positive control drug chondroitin sulfate was administered with a concentration of 1000 μg / mL in water solution, and the normal control group (administered with normal saline) and the model For the control group (administered with physiological saline), the volume of each well was 3 mL, and incubated in an incubator at 28° C. for 72 hours. After the experiment, 10 zebrafish were randomly selected in each group to observe, take pictures and save the pictures under the microscope. Nikon NIS-Elements ...

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Abstract

The invention discloses a peptide KAI 11 for promoting cartilage regeneration. The peptide KAI 11 has 9 / 11 or above sequence consistency with an amino acid sequence represented by SEQ ID NO.1. The peptide KAI 11 is derived from BMP-7, can remarkably promote cartilage regeneration, and has the advantages of easiness in synthesis, and low immunogenicity, so that the above polypeptide has wide application prospects in cartilage injury repair and osteoarthritis treatment.

Description

technical field [0001] The invention relates to the technical field of polypeptides, in particular to a peptide KAI 11 for promoting cartilage regeneration and its application. Background technique [0002] Osteoarthritis (OA) is a common joint disease, manifested as joint pain and stiffness, and cartilage damage is the main cause of osteoarthritis. Primary disease occurs frequently after middle age, and women are more than men, and the prevalence rate of 40-year-old crowd is 10%-17%, 50% over 60 years old, and up to 80% in 75-year-old crowd. The disability rate of the disease can be as high as 53%. With the aging population, osteoarthritis will become an important issue affecting people's quality of life, and the market demand for osteoarthritis drugs will continue to expand. [0003] At present, clinically, the therapeutic drugs for osteoarthritis are divided into specific therapeutic drugs and non-specific therapeutic drugs. Non-specific therapeutic drugs are mainly us...

Claims

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Application Information

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IPC IPC(8): C07K14/51A61K38/18A61P19/02A61P19/08
CPCC07K14/51A61K38/1875A61P19/02A61P19/08
Inventor 宋燕许元生孟鹤卢肖宇
Owner LINK HEALTH GRP
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