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Vaginal secretion derived polypeptide and application thereof in inhibition of candida albicans infection

A technology for Candida albicans and vagina, applied in the field of vaginal secretion-derived polypeptides and its application in inhibiting Candida albicans infection, can solve the complex problems of vaginal secretion components and mechanism of action, and achieve the goal of overcoming fungal drug resistance, molecular weight Low, infection-inhibiting effect

Active Publication Date: 2020-10-30
NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are nearly a thousand kinds of antibacterial endogenous polypeptides in vaginal secretions, and they are widely affected by hormone levels (follicular phase, luteal phase, menstrual period, pregnancy, perimenopause, etc.), which provides a basis for understanding the function of vaginal secretions. A certain molecular basis has been established, but the composition and mechanism of vaginal secretions are complex, and it is an effective way to continue to explore new protective components and mechanisms.

Method used

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  • Vaginal secretion derived polypeptide and application thereof in inhibition of candida albicans infection
  • Vaginal secretion derived polypeptide and application thereof in inhibition of candida albicans infection
  • Vaginal secretion derived polypeptide and application thereof in inhibition of candida albicans infection

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Embodiment 1

[0031] The amino acid sequence synthesized by solid phase method is alanine-threonine-proline-threonine-serine-threonine-leucine-arginine-threonine-alanine-proline- Polypeptide PDMAC of Proline-Proline-Lysine-Valine-Leucine-Threonine (ATPTSTLRTAPPPKVLT).

[0032] Chemically modify the main chain group and side chain group of the polypeptide, and use polyethylene glycol (Polyethyleneglycol, PEG) and carbamic acid to modify the C-terminal and N-terminal of the polypeptide PDMAC respectively, so as to improve the activity and stability of PDMAC, inhibit Enzymatic degradation reduces antigenicity and prolongs half-life in vivo.

[0033] The synthetic polypeptide PDMAC was added to the culture medium of vaginal epithelial cells VK2 / E6E7 to create the polypeptide action group and the control group. After incubation for 24 hours, Candida albicans was added to the polypeptide action group, and PI / SYTO9 was used for staining and flow cytometry. Cytometer technique was used to detect t...

Embodiment 2

[0037] Purchase female BALB / c mice aged 6-8 weeks at SPF level, keep a light and dark environment for 12 hours a day, control the temperature at 25°C, and feed them with food and water randomly.

[0038] Pretreatment with subcutaneous injection of 50 μg estradiol (dissolved in 100 μL filtered sterile sesame oil) 9 days before infection of the vagina and 2 days before infection, and 20 μL of Candida albicans (10 9 CFU / mL) was inoculated into the vagina of mice with an insertion depth of about 1-1.5cm. After injection, the bacterial solution was retained for about 2 hours, once a day. After 7 days, the vaginal secretions of the mice were taken for microscopic examination and biochemical pathological examination.

[0039] The results showed that after being inoculated with Candida albicans, the vulva of the mice was swollen and secretions increased; image 3 The HE staining in the middle panel a showed that the interstitium of the vaginal tissue of the mice inoculated with Candi...

Embodiment 3

[0043] Utilize the website of PredictProtein (www.predictprotein.org) to analyze the secondary structure of PDMAC, the results show that there are multiple amino acids with protein binding ability and multiple sites interacting with proteins (such as Figure 4 shown), suggesting that the function of PDMAC may be realized by interacting with the target protein.

[0044] Use the pull-down method to screen protein molecules that interact with PDMAC polypeptides (such as Figure 5 show). After biotin labeling, co-incubation with cell protein lysate, eluent elution, SDS-PAGE gradient gel separation, and silver staining, differential bands were taken for mass spectrometry analysis, and three peptides with large differences were found—— β-Tubulin, SLP-2, Beclin-1, and the mass spectrometry signal is strong.

[0045] Then PDMAC was transferred into VK2 / E6E7 cells, and the localization of PDMAC and Beclin-1 was observed by immunofluorescence experiment.

[0046] combine Figure 6 T...

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Abstract

The invention discloses a vaginal secretion derived polypeptide and an application thereof in inhibition of candida albicans infection, relates to the technical field of biology, and provides a novelpolypeptide PDMAC for inhibiting candida albicans from infecting vaginal epithelial cells. The amino acid sequence of the polypeptide is ATPTSTLRTAPPPKVLT, and the amino acid sequence of the polypeptide is ATPTSTLRTAPPPKVLT, wherein the theoretical molecular weight is 1751.06 Da, the theoretical isoelectric point is 11, the instability coefficient is 49.74, the fat coefficient is 74.71, and the hydrophilicity is -0.21, synthesizing through a solid phase method, and respectively carrying out chemical modification at the C end and the N end through polyethylene glycol and carbamic acid.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a vaginal secretion-derived polypeptide and its application in inhibiting Candida albicans infection. Background technique [0002] Vulvovaginal candidiasis (VVC) is one of the most common female vulvovaginal inflammations. Statistics show that about 75% of women have suffered from VVC at least once in their lives, and 45% of women have experienced two or more episodes. Suffering from VVC for a long time will not only bring great troubles to the life of the patient, but also lead to diseases such as cervicitis and cervical erosion, salpingo-oophoritis, pelvic inflammatory disease, secondary infertility, and malignant tumors of the reproductive tract; Risks of premature rupture of membranes, premature birth, fetal intrauterine infection, neonatal thrush, etc. Therefore, the prevention and treatment of VVC is the top priority in the research of female reproductive tract infection. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C07K1/107C07K1/04A61K38/10A61P31/10
CPCC07K7/08A61P31/10A61K38/00
Inventor 曾新李萍栾婷王新艳芮璨范裕如曾玉范翀
Owner NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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