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Polypeptide for inhibiting proliferation of candida albicans and application of polypeptide in mouthwash

A technology of Candida albicans and mouthwash, applied in peptides, oral care, antifungal and other directions, can solve problems such as large toxic and side effects, many adverse reactions, increased drug resistance reports, etc., and achieve the effect of inhibiting proliferation and preventing oral ulcers

Active Publication Date: 2021-11-26
GENERAL HOSPITAL OF NINGXIA MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although new antifungal drugs are constantly coming out, which provides new opportunities for antifungal treatment, the existing anti-Candida albicans drugs have disadvantages such as large toxic side effects and many adverse reactions, and reports of drug resistance are gradually increasing

Method used

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  • Polypeptide for inhibiting proliferation of candida albicans and application of polypeptide in mouthwash
  • Polypeptide for inhibiting proliferation of candida albicans and application of polypeptide in mouthwash
  • Polypeptide for inhibiting proliferation of candida albicans and application of polypeptide in mouthwash

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Screening of polypeptides binding to Candida albicans

[0030] (1) Amplification and purification of phage library: Inoculate a single colony of E.coli ER2738 in 10mL LB liquid medium (10g peptone, 5g yeast extract, 5g NaCl per liter, autoclave, store at room temperature), 37°C , 200rpm shaker and incubate to mid-log phase (OD 600 ≈0.5); add 10 μL of phage, shake at 37°C and 200 rpm for 4-5 hours, centrifuge at 10,000 g for 10 min, and take the supernatant; centrifuge again at 10,000 g for 10 min, take 80% of the supernatant, add 1 / 6 volume of PEG 8000 / NaCl (20 %(w / v) PEG-8000, 2.5mol / LNaCl, autoclaved, stored at room temperature), let stand overnight at 4°C. Take it out the next day, and the white precipitate is the phage. Centrifuge at 10000g for 15min, discard the supernatant, and gently suck out the residual solution after brief centrifugation; add 1mL TBS (50mmol / L Tris-HCl (pH 7.5), 150mmol / LNaCl, autoclaved, store at room temperature) solution to dissolve the w...

Embodiment 2

[0037] Identification of Candida albicans Specific Phage

[0038] Randomly pick 20 well-separated single colonies on the titer determination plate after the third round of screening in Example 1, and after culturing and purifying separately, use phage ELISA to detect the binding activity of each strain of phage to Candida albicans. The specific steps are as follows:

[0039] Coat ELISA plates with Candida albicans and Blocking respectively, each group has three parallels, and block overnight at 4°C, wash 6 times with TBST, add 100 μL of purified phage, incubate with shaking at 37°C for 1 hour, wash 10 times with TBST to remove untreated bacteria. For the bound phage, add 100 μL of HRP-labeled mouse anti-M13 monoclonal antibody, incubate at 37°C for 1 hour with shaking, wash 10 times with TBST, add TMB substrate for 10 minutes at room temperature in the dark, stop the reaction with 2 mol / L sulfuric acid, and measure the OD value at a wavelength of 450 nm , with P / N ≥ 2.1 as po...

Embodiment 3

[0042] Binding Activity Determination of Peptide Sequences

[0043] Coat the plates with different concentrations of Candida albicans and Blocking, block overnight at 4°C, wash 6 times with TBST, and add 2×10 phages with peptide A and peptide B sequences, respectively. 11 (with the same titer of phage without polypeptide A and polypeptide B sequences as the control), wash 10 times with TBST to remove unbound phage, add HRP-labeled mouse anti-M13 monoclonal antibody 100 μL, shake and incubate at 37°C for 1 hour, wash with TBST 10 times; add TMB to react at room temperature in the dark for 10 minutes, stop the reaction with 2mol / L sulfuric acid, and measure the OD value at a wavelength of 450nm. The result is as figure 2 As shown, the phages with polypeptide A and polypeptide B sequences exhibited specific binding to Candida albicans, respectively.

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Abstract

The invention provides a polypeptide for inhibiting proliferation of candida albicans and application of the polypeptide in mouthwash, and belongs to the technical field of mouthwash. The polypeptide for inhibiting proliferation of the candida albicans comprises a polypeptide A with an amino acid sequence shown as Lys-Arg-Leu-Phe-Lys-Lys-Leu and a polypeptide B with an amino acid sequence shown as Asp-Leu-Ile-Trp-Lys-Leu-Leu. The polypeptide can be specifically combined with the candida albicans, the function of the polypeptide can be closed, and proliferation of the candida albicans is inhibited. The polypeptide is applied to the mouthwash, the antibacterial and anti-inflammatory effects of the mouthwash can be further improved, and oral candida albicans infection can be prevented.

Description

technical field [0001] The invention belongs to the technical field of mouthwash, and in particular relates to a polypeptide for inhibiting the proliferation of Candida albicans and its application in mouthwash. Background technique [0002] Oral mucosal disease, also known as soft tissue oral disease, is described as a family of diseases or conditions that affect the oral mucosa and soft tissues. The most common and difficult to cure oral mucosal diseases is recurrent oral ulcers caused by oral candidiasis (OC) infection. So far, many literatures and studies have confirmed that Candida albicans is the main pathogen of oral candidiasis infection causing recurrent oral ulcers. The current treatment for Candida albicans infection mainly revolves around the use of antibiotics, and due to the abuse of antibiotics in recent years, many antibiotics also destroy other beneficial bacteria that originally colonized the oral cavity or respiratory tract in the process of inhibiting Ca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06A61K8/64A61K8/67A61P1/02A61P29/00A61P31/02A61P31/10A61Q11/00
CPCC07K7/06A61K8/64A61Q11/00A61P31/02A61P31/10A61P29/00A61P1/02A61K8/676A61K8/67A61K2800/5922Y02A50/30
Inventor 薛頔林雪张雯王玉炯
Owner GENERAL HOSPITAL OF NINGXIA MEDICAL UNIV
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