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Hydroxycholesterol 25HC and use thereof

A technology of hydroxycholesterol and use, which is applied in the field of preparation of anti-enterovirus drugs, and can solve the problems of no 25-hydroxycholesterol anti-enterovirus and no approved safe antiviral therapy

Active Publication Date: 2017-11-14
INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no specific vaccine against EV68, and there is no approved safe antiviral therapy that can be used in clinical treatment. There is an urgent need to develop specific and effective preventive and therapeutic drugs to curb the harm of the virus
However, there is no report on the anti-enterovirus of 25-hydroxycholesterol

Method used

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  • Hydroxycholesterol 25HC and use thereof
  • Hydroxycholesterol 25HC and use thereof
  • Hydroxycholesterol 25HC and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 125

[0088] The influence of embodiment 1.25HC on the replication of EV71 virus

[0089] RD cells (human rhabdomyosarcoma cells, purchased from American type culture collection) were treated with different concentrations of 25HC (H1015, purchased from American sigma company), and then inoculated with EV71 virus (Beijing strain BJ / CHN / 2008). The mRNA level of EV71 was detected, and the expression level of structural protein of EV71 virus was detected by Western blotting. The experiment was repeated 3 times, and the experimental results were averaged. The specific method is as follows:

[0090] 1.1 Virus amplification

[0091] Take 30 μl of EV71 virus and 10% FBS (fetal bovine serum, purchased from American Hyclone Company, product number 30396) in DMEM medium (purchased from American Gibco Company, product number: C11965500) and mix and inoculate in T75 (growth area 75cm 2 ) in RD cells with 80%-90% abundance (cell fraction per unit area, that is, the extended density of cells in...

Embodiment 2

[0123] Example 2. Effect of adding 25HC on EV71 virus replication at different time points of infection

[0124] Different concentrations of 25HC were added to RD cells before, at the same time and after infection with EV71 virus, and the mRNA level of EV71 was detected by fluorescent quantitative PCR. The experiment was repeated 3 times, and the experimental results were averaged. The specific method is as follows:

[0125] 1 h before inoculation of 1MOI EV71 virus (i.e. before infection), at the same time of inoculation (i.e. at the same time as infection), and 1 h after inoculation (i.e. after infection), 6.25 μM or 12.5 μM 25HC were added to RD cells respectively, and each group of experiments were set to A control without 25HC was inoculated with virus. The virus inoculation method is the same as in Example 1.3. Each group was incubated at 37°C in 5% CO after infection 2 After incubation for 24 h, discard the medium and harvest the cells. For each group of harvested ...

Embodiment 3

[0127] The influence of embodiment 3.25HC on EV68 viral replication

[0128] RD cells were treated with different concentrations of 25HC and then inoculated with EV68 virus (GeneBank number: KF726085.1), RSV virus (RSV-A2 strain virus, purchased from ATCC, USA) or EV71 virus, and the mRNA levels of different viruses were detected by fluorescent quantitative PCR. The experiment was repeated 3 times, and the experimental results were averaged. The specific method is as follows:

[0129] 3.1 Virus amplification

[0130] The amplification method of EV68 virus is the same as that of EV71, as described in Example 1.1. The amplification method of RSV virus is as follows:

[0131] HEp-2 cells (human laryngeal carcinoma cells, purchased from ATCC, number: ATCC CCL-23, cell concentration 1.8×10 5 / ml) inoculated on 75cm 2 Cell culture flasks. CO at 37°C 2 Culture in an incubator until the cell confluency reaches 60-70%. Discard the original medium, and wash once with 3ml Opti-ME...

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PUM

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Abstract

The invention discovers that 25-hydroxycholesterol is capable of effectively inhibiting copy of a 71-type enterovirus and a 68-type enterovirus. The invention provides 25-hydroxycholesterol or pharmaceutically acceptable salt or ester or use of 25-hydroxycholesterol in preparation of a drug for preventing and / or treating diseases and / or symptoms caused by the infection of enteroviruses and further provides an administration route and dose form applicable to the drug, other active components containable in the drug and combined medication of the drug and other drugs. Besides, the invention further provides a non-therapeutic purpose method for inhibiting the enteroviruses in cells by virtue of 25-hydroxycholesterol or pharmaceutically acceptable salt or ester.

Description

technical field [0001] The invention belongs to the fields of biotechnology and chemistry, and particularly relates to the use of 25-hydroxycholesterol for preparing anti-enterovirus drugs. Background technique [0002] The genus Enterovirus belongs to Picornaviridae, a class of single-stranded positive-sense RNA viruses with similar biological properties, without envelope, and with a genome length of about 7.2-8.4kb. Enteroviruses above the respiratory tract, throat and intestinal tract are the portals of invasion, and first proliferate in the local mucosa, pharynx, tonsil and other lymphoid tissues and intestinal collection lymph nodes, and then release into the blood, forming the first viremia and spreading to the local area. The target tissue with receptors will proliferate again, causing the second viremia and clinical symptoms. Receptors for most enteroviruses are widely distributed in tissues and cells, including the nervous system, heart, lung, pancreas, mucous memb...

Claims

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Application Information

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IPC IPC(8): A61K31/575A61P31/14
CPCA61K31/575Y02A50/30
Inventor 王健伟雷晓波张珍珍肖霞
Owner INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI
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