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Bone remodeling regulating polypeptides and application

A technology for regulating and osteoporosis, which is applied in the direction of peptides containing affinity tags, peptides containing His tags, peptides, etc., to achieve the effects of simple synthesis, low cost and moderate half-life

Pending Publication Date: 2020-09-11
THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no non-hormone-based peptide drugs at home and abroad

Method used

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  • Bone remodeling regulating polypeptides and application
  • Bone remodeling regulating polypeptides and application
  • Bone remodeling regulating polypeptides and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Determine the expression of collagen receptor OSCAR on osteoclasts and osteoblasts.

[0056] 1) Aseptically isolate the bilateral femur and tibia of wild-type female mice, and repeatedly wash the bone marrow cavity with the culture medium to obtain bone marrow cells. After culturing in complete α-MEM medium containing M-CSF (30ng / mL) for 24 hours, aspirate the unpasted After centrifugation, the parietal cells were resuspended, and then cultured in α-MEM complete medium containing M-CSF (30ng / mL) for 3 days to obtain adherent mouse primary bone marrow mononuclear macrophages (osteoclast precursors) cells), M-CSF (50ng / mL) and RANKL (100ng / mL) induced differentiation into osteoclasts, which were identified by TRAP staining (figure 1 A), by OSCAR immunofluorescence ( figure 1 B) and Western blot experiment ( figure 1 C) determining that osteoclast precursor cells and osteoclasts express the collagen receptor OSCAR;

[0057] 2) Aseptically isolate the skull of wild-type m...

Embodiment 2

[0059] to humans ( figure 2 A) and mice ( figure 2 B) Multiple sequence alignment of type Ⅰ collagen α1 subunit and α2 subunit to find the sequence on type Ⅰ collagen that may bind to the collagen receptor OSCAR ( figure 2 A and B box parts, the upper sequence in each group of sequences represents Col1a1, and the lower sequence represents Col1a2). And by predicting the protein structure of mouse OSCAR (mOSCAR), we found that there are 5 highly conserved amino acid sites that can bind to collagen molecules ( figure 2 C).

Embodiment 3

[0061] Based on the prediction of physical and chemical properties and functions (isoelectric point, molecular weight, number of positive and negative charges, half-life, instability index, average hydrophilicity, fatty amino acid index), screen 4 sequences derived from type Ⅰ collagen with stable performance and 1 sequence Sequences derived from type II and III collagens were synthesized from the screened sequences derived from type I collagen and sequences derived from type II and III collagens by solid-phase peptide synthesis (ie, five bone remodeling regulatory polypeptides were synthesized).

[0062] 1) By analyzing the hydroxylation level of amino acids in the protein sequence of mouse type Ⅰ collagen that may bind to the collagen receptor OSCAR ( image 3 A), and analyze the isoelectric point of the above sequence, the number of positive and negative charge residues, the number of atoms, the half-life, the instability coefficient, the average hydrophilicity and the aliph...

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PUM

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Abstract

The invention provides bone remodeling regulating polypeptides and application. At the early stage, on the basis of first experiment findings, through sequence alignment, structural analysis, physicochemical properties and function prediction, a solid-phase polypeptide synthesis method is designed and utilized to newly synthesize the bone remodeling regulating polypeptides. The polypeptides have the characteristics of short length, simple synthesis, low cost, lower cytotoxicity, higher biological stability, moderate half-life period, good targeting ability and high bone absorption regulation capacity, differentiation and functions of osteoclasts and osteoblasts can be controlled by regulating the concentration of the synthesized bone remodeling regulating polypeptides to realize ordered regulation of bone absorption and bone formation, and the polypeptides have broad potential application prospects.

Description

technical field [0001] The invention belongs to the technical field of bone remodeling regulation, and specifically relates to a bone remodeling regulating polypeptide and its application. Background technique [0002] The development and therapeutic targets of existing anti-osteoporosis drugs are almost all focused on bone remodeling (the dynamic balance between bone formation and bone resorption), that is, how to inhibit bone resorption or promote bone formation. Clinically, anti-osteoporosis drugs are also divided into anti-bone resorption drugs based on these two points, such as bisphosphonates, calcitonin, etc., and bone-promoting drugs such as teriparatide; however, the former can inhibit bone resorption. At the same time, it also inhibits bone formation, which significantly increases the incidence of adverse reactions in its long-term application; while the latter promotes bone formation and bone resorption, it may increase the incidence of fractures after 2 years of ...

Claims

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Application Information

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IPC IPC(8): C07K7/06A61K38/08A61P19/08A61P19/10
CPCC07K7/06A61P19/08A61P19/10A61K38/00C07K14/78C07K2319/21
Inventor 周后德郭玥周映辉岳海青
Owner THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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