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Nilotinib as a drug for the treatment of dengue virus infection and its pharmaceutical use

A technology of dengue virus and nilotinib, which is applied in the directions of antiviral agents, medical preparations containing active ingredients, pharmaceutical formulas, etc., can solve the problem of no dengue virus and other problems

Inactive Publication Date: 2020-05-22
SOUTHERN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although many active molecules against the main protein of dengue virus have been found, due to various reasons (such as problems such as toxic and side effects), there is no drug against dengue virus at present. Therefore, the discovery of antiviral active molecules against dengue has important biological significance. Research implications and practical implications to support the development of effective drugs against dengue virus

Method used

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  • Nilotinib as a drug for the treatment of dengue virus infection and its pharmaceutical use
  • Nilotinib as a drug for the treatment of dengue virus infection and its pharmaceutical use
  • Nilotinib as a drug for the treatment of dengue virus infection and its pharmaceutical use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Nilotinib toxicity test to BHK-21 cells

[0020] BHK-21 cells (baby hamster kidney cells) are DENV2 susceptible cells. The BHK-21 cells used in the experiment belonged to our department; MTT was purchased from Beyontian Institute of Biotechnology; fetal bovine serum was purchased from American GIBICO Company; cell culture plates were purchased from American Corning Company; RPMI 1640 medium was purchased from American GIBICO Company.

[0021] The experimental steps are as follows:

[0022] 1) Inoculation of BHK-21 cells: use RPMI 1640 medium containing 10% (V / V) fetal bovine serum to prepare a single cell suspension, and inoculate 10,000 cells per well into a 96-well cell culture plate;

[0023] 2) Culture BHK-21 cells: at 37°C, 5% (V / V) CO 2 Cultivate under culture conditions for 24 hours;

[0024] 3) Add Nilotinib: Aspirate and discard the medium in each well, add 100 μl of Nilotinib diluted to the corresponding concentration with the RPMI 1640 medium of ...

Embodiment 2

[0032] Example 2 Nilotinib inhibits DENV2 infection of BHK-21 cells:

[0033] BHK-21 is used as the cell for culturing virus DENV2, 10TCID50, the experimental steps are as follows:

[0034] 1) Add BHK-21 to the cell culture plate. After 24 hours, the cells grow to a monolayer, and the cells cover about 80% to 90% of the bottom of the well. Aspirate the medium, wash once with PBS, and insert 200 μl of virus sample , adsorption at 37°C for 1 hour. After the adsorption was completed, the virus liquid in each well was discarded and washed once with PBS. Add the specified concentration of nilotinib diluted in RPMI 1640 medium containing 10% (V / V) fetal bovine serum, and store at 37°C in 5% (V / V) CO 2 cultured under culture conditions. After 96 hours, after the cells showed obvious cytopathic changes, the cytopathic effects were observed under a microscope. See figure 1 .

[0035] 2) Collect the supernatant and measure the content of lactate dehydrogenase (LDH) released by the...

Embodiment 3

[0042] Example 3 Inhibition test of nilotinib on DENV2 key protein NS3 and E protein

[0043] 1) Collection of total cell protein: After BHK-21 cells were treated with drugs and dengue virus (DENV2), the protein was extracted for 48 hours and collected in a 1.5ml Eppendorf tube.

[0044] 2) Detection of sample protein concentration: the protein standard was diluted with double distilled water to a gradient concentration of 0, 0.0008, 0.0016, 0.0032, 0.004, 0.006, 0.008mg / ml; the protein concentration in the sample was calculated according to the standard curve. Make the protein concentration consistent for each sample.

[0045]3) Western blot detection of dengue virus protein expression differences after gradient concentration nilotinib treatment: add various reagents in order, and start electrophoresis with 80V constant voltage for electrophoresis. After the front of the dye enters the separating gel, change 120V, according to the degree of separation of the pre-stained prot...

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Abstract

The invention relates to nilotinib as a medicine for treating dengue virus infection and a pharmaceutical application thereof and particularly discloses an application of the nilotinib in preparing the medicine for treating or preventing dengue virus infection. The nilotinib and the pharmaceutical application have the beneficial effects that proved by inhibition test for DENV, the nilotinib has good inhibitory action for DENV2 so as to prove that that the medicine has wide application prospect in treating a DENV2 infection disease fundamentally. The nilotinib can provide good candidate medicine for clinically treating the diseases caused by DENV2, and has good application prospect.

Description

technical field [0001] The invention relates to the field of medicines, in particular to the use of nilotinib in resisting dengue fever virus. Background technique [0002] Dengue virus (dengue virus, DENV) belongs to Flaviviridae, Flaviviridae, is the most widely distributed arbovirus in the world today, mainly caused by Aedes aegypti and Aedes albopictus that bite people Dengue fever, among which dengue hemorrhagic fever / dengue shock syndrome (dengue hemorrhagic fever / dengue shock syndrome, DHF / DSS) is more serious. This virus has four serotypes, and serotype 2 is an epidemic strain. Dengue fever (denguefever) is an acute infectious disease transmitted by mosquitoes, and is mainly prevalent in tropical and subtropical regions. Since dengue fever was first discovered in 1779, dengue fever outbreaks have occurred in various parts of the world. According to WHO estimates, about 2 / 5 of the world's population is threatened by dengue fever, and hundreds of millions of people a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/506A61P31/14
CPCA61K31/506Y02A50/30
Inventor 姚新刚刘叔文
Owner SOUTHERN MEDICAL UNIVERSITY
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