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Novel parathyroid hormone (PTH) mimetic peptide based on protein domain reconstruction, and application of novel PTH mimetic peptide

A protein domain, peptide-mimicking technology, applied in the direction of peptide/protein components, animal/human proteins, medical preparations containing active ingredients, etc. Effect

Active Publication Date: 2019-02-12
北京兆来医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the recent study of sclerostin antibody, it was found that if the bone resorption element in bone metabolism is inhibited, the effect of promoting osteogenesis will not last long, because bone synthesis and bone resorption are two complementary links in the bone metabolism process.

Method used

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  • Novel parathyroid hormone (PTH) mimetic peptide based on protein domain reconstruction, and application of novel PTH mimetic peptide
  • Novel parathyroid hormone (PTH) mimetic peptide based on protein domain reconstruction, and application of novel PTH mimetic peptide
  • Novel parathyroid hormone (PTH) mimetic peptide based on protein domain reconstruction, and application of novel PTH mimetic peptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 MY-3 has a stronger effect on promoting osteoblast differentiation than PTH (1-34) during intermittent stimulation, and both have the same effect on promoting osteoclast differentiation

[0027] Osteoblasts were isolated from neonatal mouse skull and cultured in vitro. 1x10 5 The density per well was inoculated in a 24-well culture plate, and the 4h / 48h intermittent PTH mimic peptide stimulation experiment was started after 3 days, that is, the PTH mimic peptide was added to the culture system (containing 10% serum+α-MEM medium) at a corresponding concentration, 4 Aspirate the culture medium after 1 hour, gently wash the adherent cells once with α-MEM medium, replace with 10% serum + α-MEM medium and continue culturing for 44 hours; the cycle repeats. Concentration: MY-1 Peptide 10 -6 M, PTH(1-34) peptide 10 -8 M, MY-3 peptide 10 -8 M.

[0028] Alkaline phosphatase staining was performed 2 weeks later, and calcium nodule staining was performed 4 weeks lat...

Embodiment 2

[0033] Example 2: When continuously stimulated, MY-3 has a weaker effect on inhibiting osteoblast differentiation than PTH (1-34), and both have the same effect on promoting osteoclast differentiation

[0034] Osteoblasts were isolated from neonatal mouse skull and cultured in vitro. 1x10 5 Inoculate in a 24-well culture plate at a density of 24 wells. After 3 days, start the continuous PTH mimic peptide stimulation experiment. Add the PTH mimic peptide to the culture system (containing 10% serum + α-MEM medium) at the corresponding concentration, and replace the culture after 48 hours. Add new PTH mimetic peptides at base time, that is, PTH mimetic peptides are always in the culture solution. Concentration: MY-1 Peptide 10 -6 M, PTH(1-34) peptide 10 -8 M, MY-3 peptide 10 -8 M. Alkaline phosphatase staining was performed 2 weeks later, and calcium nodule staining was performed 4 weeks later ( figure 2 A, the upper two rows of holes). And quantitatively detect the expre...

Embodiment 3

[0039] Example 3: MY-3 has a stronger osteogenic gene expression effect than PTH (1-34), and both have the same expression effect of osteoclast differentiation genes

[0040] Primary mouse calvarial osteoblasts ( image 3 A and image 3 B) and mouse bone marrow cells ( image 3 C and image 3 D) Cultured in cell culture plates. When the cell growth completely covered the bottom of the culture well, the culture medium containing 1% fetal calf serum was used instead, and different concentrations of PTH mimic peptides (concentrations as described above) were added, and the cells receiving intermittent stimulation ( image 3 A and image 3 C) The method of using PTH peptide for 4 hours and replacing with ordinary medium for 44 hours, a total of three 48-hour cycles. Cell RNA was extracted 4 hours after the last PTH peptide stimulation for Realtime PCR detection. Cells receiving continuous stimulation ( image 3 B and image 3 D) The PTH peptide stays in the medium, and the...

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Abstract

The invention discloses novel parathyroid hormone (PTH) (1-34) mimetic peptide based on protein domain reconstruction.; The mimetic peptide contains peptide fragment repeats at 29-34th positions, andthe 34th position of the mimetic peptide and the last amino acid of the peptide fragment repeats at 29-34th positions are changed into tyrosine (Tyr, Y) from phenylalanine (Phe, F), so that MY-3 peptide is formed; preclinical experiments show that the MY-3 mimetic peptide can promote osteoblast differentiation, and has a very good effect of treating osteoporosis and promoting bone growth; the anti-osteoporosis effect of the MY-3 mimetic peptide is better than that of the PTH (1-34) (a product on the market is named as Forteo or Teriparatide), so that the MY-3 mimetic peptide has a significantclinical application prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a novel PTH mimic peptide based on protein domain reconstruction and its application. Background technique [0002] Osteoporosis is a systemic skeletal disorder characterized by low bone mass, microstructural deterioration of bone tissue, and increased bone fragility and susceptibility to fracture. A common disease that seriously affects the health and life of the elderly. At present, common drugs for the treatment of osteoporosis include anti-resorptive drugs and osteosynthetic drugs. The former cannot effectively improve bone quality and requires long-term medication. The latter can quickly improve bone quality and prevent fractures, especially for severe osteoporosis. The loosening effect is obvious. [0003] Parathyroid hormone (PTH) is a polypeptide hormone secreted by the parathyroid gland and is an important factor in regulating calcium and phosphorus metabolic balance in orga...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/635A61K38/29A61P19/10A61P19/08
CPCA61K38/00A61P19/08A61P19/10C07K14/635
Inventor 杨德鸿
Owner 北京兆来医药科技有限公司
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