A kind of anti-calcification short peptide and its application

A technology of short peptides and drugs, which is applied in the field of biomedicine to achieve the effect of short length and easy synthesis

Active Publication Date: 2020-05-26
孙伟
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is a pity that there is no effective clinical method for preventing or treating calcification at present. The current treatment can only use artificial organs to replace severely calcified tissues, such as artificial valves to replace calcified heart valves, and artificial blood vessels to replace calcified tissues. blood vessels, etc.
For patients with renal insufficiency, there is no effective method to delay long-term dialysis-related cardiovascular calcification, and it is urgent to develop some effective treatments, especially long-term treatments, to alleviate the harm of calcification diseases

Method used

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  • A kind of anti-calcification short peptide and its application
  • A kind of anti-calcification short peptide and its application
  • A kind of anti-calcification short peptide and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1, the operation steps of the preparation method of short peptide:

[0025] The main operation steps of solid-phase short peptide are as follows:

[0026] (1) Weigh 0.5 mmol of Fmoc-Thr(tBu)wang resin with a loading of 0.35 mmol / g, place it in a reaction column, add an appropriate amount of DCM to soak for 30 minutes, remove the DCM, and wash with DMF for 3 times.

[0027] (2) For deprotection, add an appropriate amount of deprotection solution (20% piperidine in DMF) to react for 20 min, remove the deprotection solution, and wash with DMF for 6 times.

[0028] (3) Coupling of amino acids, weigh 3eq of Fmoc-Pro-OH and TBTU, add to the reaction column, use DMF as solvent, add 3eq of DIEA, react for 1h, check whether the reaction is complete by ninhydrin, after the reaction , the reaction solution was removed and washed three times with DMF.

[0029] (4) Steps (2) and (3) are repeated until the last amino acid reaction is completed.

[0030] (5) Deprotection...

Embodiment 2

[0033] Example 2, Experiment of Short Peptides Inhibiting Warfarin-Induced Valvular Interstitial Cell Calcification

[0034] Primary porcine aortic valve mesenchymal cells were cultured in DMEM medium containing 10% fetal bovine serum at a cell density of about 1*10 5 The amount of cells per well was added to a 12-well plate, and 1.6mM warfarin solution was added to culture for 7 days. At the same time, different concentrations of the short peptide of the present invention and the negative control peptide (SEQ ID NO.3) were added respectively. The cells were fixed with 4% paraformaldehyde and stained with 1% alizarin red to show the calcium deposition. In addition, 0.6M dilute hydrochloric acid was used to dissolve intracellular calcium, and the colorimetric calcium concentration determination kit was used to detect intracellular calcium. levels of calcium content.

[0035] The results show that the short peptide of the present invention can significantly inhibit warfarin-ind...

Embodiment 3

[0036] Example 3, Experiment of Short Peptides Inhibiting Calcification of Valve Interstitial Cells Induced by High Calcium and High Phosphorus

[0037] Primary porcine aortic valve mesenchymal cells were cultured in DMEM medium containing 10% fetal bovine serum at a cell density of about 1*10 5 The amount of cells per well is added to a 12-well plate, 2.0mM calcium chloride and 2.0mM (sodium hydrogen phosphate + disodium hydrogen phosphate) solution are added for 5 days, and different concentrations of short peptides and negative peptides of the present invention are added respectively. For the control peptide (SEQ ID NO.3), the cells were fixed with 4% paraformaldehyde on day 5 and stained with 1% alizarin red to show the calcium deposition, and the intracellular calcium was dissolved with 0.6M dilute hydrochloric acid , using a colorimetric calcium concentration assay kit to detect the level of intracellular calcium content. The results show that the short peptide of the p...

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PUM

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Abstract

The invention relates to an anti-calcification oligopeptide, in particular to the anti-calcification oligopeptide and application thereof. The amino acid sequence of the anti-calcification oligopeptide is shown in SEQ ID NO.1. It is proved through experiments that the oligopeptide can obviously suppressing valvular interstitial cell calcification induced by Warfarin, high calcium and high phosphorus, and can suppress valvular interstitial cell apoptosis induced low-glucose culture. Therefore, the oligopeptide potentially becomes a good low-side-effect anti-calcification drug.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular to an anti-calcification short peptide and its application. Background technique [0002] Calcification disease is a very common clinical disease that occurs in non-bone tissue similar to the osteogenesis process and is mainly manifested by calcium salt deposition. Calcification most often occurs in heart valves, large blood vessels, synovium, tracheal cartilage, etc. Elastic soft tissue. In recent years, more and more studies have shown that ectopic calcification, such as the most common heart valves and blood vessels, is an active regulation process under the joint action of extrinsic and intrinsic factors. At present, the main clinical causes of ectopic calcified lesions are chronic inflammatory diseases such as long-term atherosclerosis, calcium and phosphorus metabolism disorders caused by renal insufficiency, calcium and phosphorus metabolism disorders caused by endocrine diseases,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/81A61K38/57A61P9/00A61P9/10A61P13/12A61P11/00
CPCA61K38/00C07K14/81
Inventor 孙伟孔祥清邱铭陆艳纪玥
Owner 孙伟
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