Application of triptolide in preparation of medicament for treating or preventing human immunodeficiency viruses (HIV)

A technology of triptolide, HIV, applied in the field of medicine

Inactive Publication Date: 2012-10-31
WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the anti-HIV-1 activity

Method used

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  • Application of triptolide in preparation of medicament for treating or preventing human immunodeficiency viruses (HIV)
  • Application of triptolide in preparation of medicament for treating or preventing human immunodeficiency viruses (HIV)
  • Application of triptolide in preparation of medicament for treating or preventing human immunodeficiency viruses (HIV)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Cytotoxicity and anti-HIV-1 activity of triptolide in TZM-b1 cells

[0026] In this example, HIV-1 NL4-3 was used to infect TZM-b1 cells (from the National Institute of Allergy and Infectious Diseases). TZM-b1 cells stably express the HIV-1 receptor CD4, and the co-receptors CXCR4 and CCR5, and integrate the reporter gene firefly luciferase controlled by the HIV-1 promoter LTR in the genome. The expression level of the reporter gene after virus infection can indicate the level of virus replication. In the experiment, Cell TiterGlo (product of Promega) was used to quantitatively detect the cell viability to determine the cytotoxicity of triptolide, and SteadyGlo (product of Promega) was used to detect the expression level of the reporter gene in the infected cells. The specific experimental method is as follows:

[0027] Cytotoxicity experiment: trypsinize TZM-b1 cells in the logarithmic growth phase, adjust the cell density to 1×10 5 / mL, 100 μL of cell su...

Embodiment 2

[0032] Example 2: Cytotoxicity and anti-HIV-1 activity of triptolide in Jurkat T lymphocytes

[0033] In this example, the applicant used the HIV-1 single-round replication system to verify the anti-HIV-1 activity of triptolide. In this system, Jurkat cells (purchased from the American Type Culture Collection) were infected with HIV-1 pseudoviruses coated with the VSV-G envelope protein. Since the viral genome has a reporter gene, the expression of the reporter gene can indicate the replication level of the virus. The experimental method is as follows:

[0034] According to the method of using Lipofectamine2000 (product of Roche Company), pNL4-3.Luc.R-E- (from the National Institute of Allergy and Infectious Diseases of the United States) and pVpack-VSV-G (product of Stratagene Company) were combined at a mass ratio of 2:1. 293T cells were transfected, and the supernatant was collected 48 hours later, filtered through a 0.45 μM filter membrane and stored at -80°C.

[0035] Ju...

Embodiment 3

[0040] Example 3: Cytotoxicity and anti-HIV-1 activity of triptolide in peripheral blood mononuclear cells (PBMCs)

[0041] In this example, the applicant detected the antiviral effect of triptolide on HIV-1NL4-3 in PBMCs, the experimental method is as follows:

[0042] Peripheral blood mononuclear cells from healthy people were separated using lymphocyte separation medium Ficoll-Hypaque PREMIUM1.073 (product of GE Company). The cells were cultured in RMPI1640 (product of Gibco) containing 50ng / mL IL-2 (product of Sigma-Aldrich) and 10% FBS (product of Gibco). Before the experiment, PBMCs were stimulated by lectin (5 μg / mL, product of Sigma-Aldrich Company) for 72 hours. HIV-1NL4-3 infected PBMCs, MOI=0.01. cells in 2 x 10 5 Inoculate in a 96-well plate at a density per well, and add appropriately diluted drugs to a total volume of 200 μL. After every 48 h of culture, 100 μL of supernatant was taken out for p24 detection, and 100 μL of fresh culture medium was added to con...

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Abstract

The invention discloses an application of triptolide in preparation of a medicament for treating or preventing human immunodeficiency viruses (HIV). The triptolide is diterpenoid naturally existing in roots of tripterygium wilfordii hook, and can dose-dependently inhibit replication of I type HIV (HIV-1) in cells in vitro. The half inhibitory concentrations on HIV-1 inhibition in TZM-b1 cells, JurkatT lymphocytes and human peripheral blood mononuclear cells are respectively 0.32nM, 0.45nM and 1.1nM. The triptolide has a remarkable inhibiting effect on the replication of the HIV-1 in the TZM-b1 cells, the JurkatT lymphocytes and the human peripheral blood mononuclear cells; and the triptolide is an active ingredient in Chinese medicinal tripterygium wilfordii hook, so the triptolide is wide in source. The compound has a broad prospect for developing anti-HIV-1 medicaments.

Description

technical field [0001] The invention belongs to the technical field of medicine, and more specifically relates to the application of triptolide in the preparation of drugs for treating or preventing HIV. Background technique [0002] AIDS, also known as acquired immunodeficiency syndrome (AIDS), is an infectious disease caused by human immunodeficiency virus (human immunodeficiency virus, HIV) and characterized by severe damage to the systemic immune system. In 2007, a total of 33 million people worldwide were living with HIV, 2.7 million newly infected people, and 2 million people died of AIDS-related diseases. The AIDS epidemic in my country is accelerating, and HIV is spreading to the general population. In 2009, 740,000 HIV-infected and AIDS patients survived. The epidemic of AIDS is a serious threat to human health and the development of the global economy. [0003] HIV is an RNA virus belonging to the family Retroviridae and the genus Lentivirus. During HIV infectio...

Claims

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Application Information

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IPC IPC(8): A61K31/585A61P31/18
Inventor 陈绪林万志涛
Owner WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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