Application of small molecule inhibitors of erk signaling pathway in inhibiting chlamydia infection

A technology of small molecule inhibitors and signaling pathways, applied in the field of biomedicine, can solve the problems of targeted host treatment of chlamydia infection that have not yet been seen

Active Publication Date: 2021-04-27
DERMATOLOGY HOSPITAL SOUTHERN MEDICAL UNIV (GUANGDONG PROVINCIAL DERMATOLOGY HOSPITAL GUANGDONG PROVINCIAL CENT FOR STI & SKIN DISEASES CONTROL & PREVENTION RES CENT FOR LEPROSY CONTROL & PREVENTION CHINA)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In recent years, adjuvant host-targeted therapy strategy is a new direction for the treatment of microbial infections. Targeted host therapy strategies have made some progress in the treatment of HIV, tuberculosis and fungal infections, but there have been no reports on assisted host-targeted therapy for chlamydial infection.

Method used

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  • Application of small molecule inhibitors of erk signaling pathway in inhibiting chlamydia infection
  • Application of small molecule inhibitors of erk signaling pathway in inhibiting chlamydia infection
  • Application of small molecule inhibitors of erk signaling pathway in inhibiting chlamydia infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 Screening of signaling pathway inhibitors that inhibit Chlamydia trachomatis infection

[0058] We screened the inhibitors of 5 signaling pathways p38 MAPK, JNK, Akt, PI3K and ERK signaling pathways, clarified the role of the 5 signaling pathway inhibitors in Chlamydia trachomatis infection, observed the infection rate of Chlamydia, the number of inclusion bodies, inclusion Body size, the number of infectious chlamydia produced after replanting, and the inhibitor with the strongest inhibitory effect was selected for further research. Research technology routes such as figure 1 shown.

[0059] The result is as figure 2 , Immunofluorescence diagram of the growth of Chlamydia trachomatis type D under the action of different inhibitors (green fluorescence: anti-MOMP) (×200) A: Negative control (no Chlamydia infection); B: Positive control (normal infection), more visible Large inclusion bodies with green fluorescence; C: 0.08 μg / ml azithromycin treatment, no i...

Embodiment 2

[0061] Example 2 VX-11e and BVD-523 can significantly inhibit the infection of Chlamydia

[0062] Two small molecule inhibitors of ERK, VX-11e and BVD-523, were selected for in-depth research, and Chlamydia trachomatis type D and human cervical cancer cells (Hela) were selected to clarify the role of VX-11e and BVD-523 in Chlamydia infection. Compared with the MEK inhibitor U0126 that has been reported in the literature, the advantages of VX-11e and BVD-523 in inhibiting Chlamydia infection are clarified. Research technology routes such as image 3 .

[0063] The specific experimental method is as follows:

[0064] 1. Experimental method

[0065] 1.1 Cell recovery and inoculation of Chlamydia: Take out the frozen Hela cells from -70°C, place them in a water bath at 37°C for shaking, dissolve them instantly, and put them in DMEM medium containing 10% newborn calf serum, 5% CO 2 , Cultured at 35°C, digested with trypsin and passaged. Hela cells were plated in 24-well plates...

Embodiment 3

[0077] Example 3 VX-11e and BVD-523 have similar effects in different cell lines

[0078] Establish mouse fibroblast McCoy and green monkey kidney Vero cell infection models to clarify the effect of VX-11e and BVD-523 on inhibiting Chlamydia infection in different cell types. The specific experimental method is as follows:

[0079] 1. Experimental method:

[0080] 1.1 Chlamydia inoculation: take out frozen McCoy cells and Vero cells from -70°C, dissolve them at 37°C, and put them in DMEM medium containing 10% newborn bovine serum, 5% CO 2 , Cultured at 35°C, until the cells grew into a dense monolayer, digested with trypsin and passaged. McCoy cells and Vero cells were plated in 24-well plates, and slides were added, 1×10 per well 5 Cells, after the cells have grown into a monolayer, are inoculated with Chlamydia. Take out the Chlamydia strain from -70°C, shake on a vortex shaker for 30 sec, and transfer the content to a monolayer cell culture plate under sterile condition...

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Abstract

The invention discloses the application of small molecule inhibitors VX-11e and BVD-523 in inhibiting chlamydia infection. The study of the present invention found that the signaling pathway inhibitors VX‑11e and BVD‑523 can significantly inhibit the infection of Chlamydia trachomatis, compared with the reported MEK inhibitor U0126, there is a significant difference; the present invention is the first small molecule inhibitor VX‑11e and BVD‑523 applied to chlamydia infection, which is expected to become a new drug for chlamydia targeted host therapy; at the same time, the study also found that the combined application of inhibitors VX‑11e and BVD‑523 with azithromycin has a synergistic effect, after chlamydia infection, VX‑11e and BVD‑523 can promote the anti-infective effect of azithromycin, and have important application value for the treatment of chlamydia infection. The invention is of great significance for the development of new drugs for chlamydia trachomatis infection and for finding new targets for assisting host treatment.

Description

technical field [0001] The invention belongs to the technical field of biomedicine. More specifically, it relates to the application of small molecule inhibitors VX-11e and BVD-523 of ERK signaling pathway in inhibiting Chlamydia infection. Background technique [0002] Chlamydia trachomatis (Ct) infection has become one of the most prevalent sexually transmitted diseases worldwide. If Ct infection is not treated in time, it can cause serious complications, such as male epididymitis and prostatitis, female cervicitis, pelvic inflammatory disease, salpingitis, ectopic pregnancy and infertility, etc.; it can also spread through the birth canal, causing neonatal eyes Conjunctivitis and pneumonia. In addition, Ct infection is also a synergistic factor for human papillomavirus-induced cervical cancer and an important synergistic factor for HIV infection. [0003] WHO, the United States and my country all recommend azithromycin and doxycycline as the first-choice treatment drug...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/00A61K45/06A61K31/7052A61K31/4439A61K31/506A61P31/00
Inventor 薛耀华郑和平荣知立
Owner DERMATOLOGY HOSPITAL SOUTHERN MEDICAL UNIV (GUANGDONG PROVINCIAL DERMATOLOGY HOSPITAL GUANGDONG PROVINCIAL CENT FOR STI & SKIN DISEASES CONTROL & PREVENTION RES CENT FOR LEPROSY CONTROL & PREVENTION CHINA)
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