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Antiviral applications of cholesterol transport inhibitor

An inhibitor, cholesterol technology, applied in the direction of antiviral agents, medical preparations containing active ingredients, steroids, etc.

Inactive Publication Date: 2019-07-12
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Whether cholesterol transport inhibitors can affect virus entry has not been reported in the prior art

Method used

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  • Antiviral applications of cholesterol transport inhibitor
  • Antiviral applications of cholesterol transport inhibitor
  • Antiviral applications of cholesterol transport inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1 Cytotoxicity detection of NPC1 protein inhibitor U18666A

[0073] 1. Experimental materials

[0074] 1.1 Cells, Compounds, Kits

[0075] PK15 (porcine kidney epithelial cells), 3D4 / 21 (porcine alveolar macrophages) and HeLa (human cervical cancer cell) cells were purchased from the American Type Culture Collection (ATCC); MARC-145 (African green monkey kidney cells) Donated by Henan Academy of Agricultural Sciences; CCK kit (ZP328) was purchased from Beijing Zhuangmeng International Biogene Technology Co., Ltd.

[0076] 1.2 Experimental Instruments

[0077] Varioskan Flash spectral scanning multifunction reader (Thermo Fisher, USA)

[0078] 2. Experimental methods and results

[0079] 2.1 Cell culture: PK15, MARC-145 and HeLa cells at 37°C, 5% CO 2 Culture in a humidified incubator, using DMEM (CORING) medium containing 10% FBS, 100U / ml penicillin and streptomycin; 3D4 / 21 cells in RPIM 1640 (CORING) containing 10% fetal bovine serum (PAN) nourish. Cells...

Embodiment 2

[0083] Example 2 Detection of U18666A cytotoxicity in mice

[0084] 1 Experimental materials

[0085] Balb / C mice (purchased from Zhengzhou University), infusion set needles, 4% PFA, 1×PBS, needles for fixing limbs (4 pieces), 2 scissors, 1 medium-sized blunt-tip tweezers, 1 small-sized tweezers Put, alcohol watering can, 2 30ml syringes, ether for anesthesia.

[0086]2 reagent formula

[0087] a) The formula of hematoxylin staining solution

[0088]

[0089] b) 1% hydrochloric acid alcohol; c) 0.5% eosin alcohol solution; d) 30% sucrose solution; e) 10% formaldehyde solution; f) 70% ethanol.

[0090] 3 experimental equipment

[0091] Fluorescence microscope (OLYMPUS)

[0092] 4 Experimental methods and results

[0093] 4.1 Body weight change and liver toxicity detection of mice treated with U18666A: 4-6 week-old experimental mice were fed normally for one week and divided into three groups (6 mice / group) according to body weight, and then the three groups were divide...

Embodiment 3

[0118] Example 3 U18666A inhibits clathrin-mediated viral endocytosis

[0119] 1. Experimental materials

[0120] 1.1 cells, plasmids, kits

[0121] PK15 (porcine kidney epithelial cells), HEK293T / 17 (human kidney epithelial cells) and HeLa (human cervical cancer cell) cells were purchased from the American Type Culture Collection (ATCC); AP2A1-FLAG, AP2A2-FLAG, AP2B1-FLAG , AP2M1-FLAG, AP2S1-FLAG, CLTA-FLAG, CLTB-FLAG, AP2A1-FLAG, AP2B1-mcherry plasmids were all constructed by our laboratory; 21aa-mcherry is a plasma membrane localization plasmid, constructed by our laboratory; EZ-Link TM Sulfo-NHS-SS-Biotin, Pierce TM Neutr Avidin TM Agarose, Transferrin From Human Serum, and Alexa Fluor488 Conjugate were purchased from Thermo Fisher Scientific; Rabbit-anti AP2B1 antibody was purchased from proteintech; 35mm glass-bottom culture dishes were purchased from NEST.

[0122] 1.2 Experimental Instruments

[0123] Amersham Imager 600 chromogenic instrument (GE Healthcare); LSM...

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Abstract

The invention relates to an application of a cholesterol transport inhibitor in preparation of a reagent for inhibiting viruses from entering cells through clathrin-mediated viral endocytosis. The invention also relates to an application of the cholesterol transport inhibitor in preparation of a medicine resistant to enveloped virus infection. The novel applications of the cholesterol transport inhibitor discovered by the invention have important significance for biology, particularly viral research and drug development.

Description

technical field [0001] The present invention relates to the antiviral use of cholesterol transport inhibitors, especially in the preparation of reagents for inhibiting clathrin-mediated viral endocytosis into cells. Background technique [0002] Cholesterol is an important component of cell membranes and an important precursor for the production of other biomolecules. Cholesterol can be obtained intracellularly in two ways: one is de novo synthesis and the other is exogenous uptake. Exogenous uptake of cholesterol comes primarily from circulating low density lipoproteins (LDL) in the blood (Brown and Goldstein 1986). Normally, cholesterol is present in the blood as cholesteryl esters through LDL particles. Although cholesterol is freely partitioned in and out of most cell membranes, it cannot be exported from endosomes / lysosomes to other membrane components without the help of NPC1 and NPC2 (Kwon, Abi-Mosleh et al. 2009, Li, Saha et al. 2016, Pfeffer 2016). [0003] Chol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07J41/00A61P31/14A61P31/16A61P31/22A61K31/56
CPCA61P31/14A61P31/16A61P31/22C07J41/0044
Inventor 褚贝贝王江杨国宇李国利明胜利
Owner HENAN AGRICULTURAL UNIVERSITY
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