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Salivary proline-rich protein bionic anti-caries functional polypeptide, and derivative, salt and application of salivary proline-rich protein bionic anti-caries functional polypeptide

A technology of peptide derivatives with multiple functions, which can be applied in the direction of peptide/protein components, peptide sources, drug combinations, etc., can solve the problems of natural salivary protein with large molecular weight, high price, and difficult extraction, and achieve good anti-caries effect and low production cost. low cost effective

Active Publication Date: 2018-01-09
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the molecular weight of natural salivary protein is large, and there are disadvantages such as difficulty in extraction, high price, and easy denaturation. Therefore, in order to achieve the purpose of preventing or blocking dental caries, it is necessary to develop and synthesize bionic anti-caries functional polypeptides with the function of promoting mineralization. important research significance

Method used

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  • Salivary proline-rich protein bionic anti-caries functional polypeptide, and derivative, salt and application of salivary proline-rich protein bionic anti-caries functional polypeptide
  • Salivary proline-rich protein bionic anti-caries functional polypeptide, and derivative, salt and application of salivary proline-rich protein bionic anti-caries functional polypeptide
  • Salivary proline-rich protein bionic anti-caries functional polypeptide, and derivative, salt and application of salivary proline-rich protein bionic anti-caries functional polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A saliva-rich protein biomimetic anti-caries functional polypeptide, the amino acid sequence of which is SEQ ID NO.1: PPGKPQGPPPQG (hereinafter referred to as PG12).

[0029] Prepare as follows:

[0030] 1. Select Fmoc-His(Trt)-Wang Resin as the resin (carrier);

[0031] 2. Fully swell the resin with DCM;

[0032] 3. Use an appropriate concentration of DBLK (hexahydropyridine + DMF) to remove the Fmoc-protecting group;

[0033] 4. Wash several times with DMF to remove DBLK;

[0034] 5. Weigh a suitable condensing agent and activator (HBTU, NMM) and the second Fmoc-protected amino acid (Fomc-Leu-OH) at the C-terminal for coupling;

[0035] 6. The ninhydrin detection method is used for detection to ensure that the connection is relatively complete;

[0036] 7. Wash several times with DMF to wash away the remaining residues and activator condensation agent;

[0037] 8. Carry out coupling according to the amino acid sequence of SEQ ID NO.1, the method refers to steps 3...

Embodiment 2

[0042] Example 2 Detection of Polypeptide Nucleation Ability to Hydroxyapatite

[0043] 1. Prepare a 50 μM polypeptide PG12 solution, and add the final concentration of 1.6mM Na 2 HPO 4 and 3.3mM CaCl 2 solution, the pH was adjusted to 7.4, and incubated on a shaker at 37°C for 24h (100 rpm).

[0044] 2. Take 10 μl of the reacted solution and add it dropwise on the copper grid. The negative control is Na without peptide. 2 HPO 4 and CaCl 2 solution. The shape of the precipitate on the copper grid was observed under a transmission electron microscope. figure 1 It shows that the crystals formed in the polypeptide PG12 group are denser than those in the negative control group, and are in the form of bundles or columns, suggesting that PG12 has a good ability to promote the nucleation and growth of hydroxyapatite.

[0045] 3. Selection of electron diffraction shows that the crystal precipitate formed by the polypeptide PG12 group has the characteristic diffraction rings 004...

Embodiment 3

[0047] Example 3 Study on the Remineralization Effect of Polypeptides on Demineralized Tooth Enamel

[0048] In this example, the remineralization effect of the polypeptide on early artificial enamel caries was observed through a static remineralization experiment.

[0049] The experimental steps are as follows:

[0050] 1. Preparation of enamel samples: freshly extracted bovine incisors were selected to prepare bovine tooth enamel samples. Under running water, use aluminum oxide paste to remove enamel surface staining, calculus and irregular surface, deionized water ultrasonic cleaning for 20 minutes, store in PBS containing 0.05% thymol, and place in a refrigerator at 4 °C spare. The crown and root were separated, and the tooth tissue of the crown was ultrasonically cleaned for 20 minutes, and then dried naturally. The crown tissue with a smooth surface, no fluorosis, no pigment, and no cracks was selected for the next step. Use a hard tissue high-speed cutting machine to...

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Abstract

The invention discloses a salivary proline-rich protein bionic anti-caries functional polypeptide, and a polypeptide derivative or a pharmaceutically acceptable salt, the amino acid sequence of the salivary proline-rich protein bionic anti-caries functional polypeptide is shown in SEQ ID NO.1, the polypeptide is one polypeptide segment of a neutral proline-rich protein 2 in saliva, has a capability of promoting the nucleation of hydroxyapatite, promotes remineralization of demineralized enamel, and reduces caries depth of the demineralized enamel and the mineral loss; the polypeptide is excellent in structure stability and has no obvious cytotoxicity, and compared with a natural proline-rich protein, the polypeptide is low in molecular weight, can be artificially synthesized, is low in cost, is easy to purify, and has a higher potential of a patent medicine. The invention further provides an application of the polypeptide to the preparation of an anti-caries medicine.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a saliva-rich protein bionic anti-caries functional polypeptide, a polypeptide derivative or a salt thereof and the application thereof in pharmacy. Background technique [0002] Dental caries is an infectious disease and the most common oral disease in humans. It has a high incidence rate and a wide range of epidemic areas, seriously affecting oral and general health. The World Health Organization has listed it as the three major non-communicable diseases that human beings should focus on one. The pathogenesis of caries is the continuous demineralization of tooth hard tissue under the action of oral cariogenic bacteria acid production. Therefore, promoting the remineralization of demineralized tooth hard tissue is an important aspect of caries prevention and treatment. [0003] As a classic anti-caries preparation, fluoride can reduce the caries rate in the population t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47A61K38/17A61P1/02
Inventor 张凌琳王琨王雨霏倪超周学东李伟
Owner SICHUAN UNIV
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