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Application of small molecule compounds in preparation of silkworm antiviral drug

A technology of small molecular compound, silkworm baculovirus, applied in antiviral agents, medical preparations containing active ingredients, pharmaceutical formulations, etc. Development did not keep up with other issues

Active Publication Date: 2019-01-18
JIANGSU UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, due to the weak research and development strength of the silkworm medicine industry and the lack of capital investment, the development of silkworm medicines, especially the development of silkworm antiviral medicines, has not kept up with the pace of other animal and plant antiviral medicines.

Method used

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  • Application of small molecule compounds in preparation of silkworm antiviral drug
  • Application of small molecule compounds in preparation of silkworm antiviral drug
  • Application of small molecule compounds in preparation of silkworm antiviral drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] 1. Construction of BmNPV carrying green fluorescent protein reporter gene

[0023] BmBacJS13 is a Bacmid strain with the same infectious properties as BmNPV (Huang JSetal. Construction of the Bac-to-Bac System of Bombyxmori Nucleopolyhedroviru. Virologica Sinica. 2007, 22(3): 218-225). In order to facilitate the observation and statistics of the control effect described in this patent, the reporter gene egfp is transposed to the Tn7 transposition insertion site of BmBacJS13, wherein egfp can adopt the sequence commonly used in this field, without special limitation, as long as it can play the role of reporter indication. Can.

[0024] The specific method of this step is as follows:

[0025] The egfp fragment was cloned into the EcorI (Takara) and XhoI (Takara) sites of pFastBac1 (Invitrogen Company), and the donor plasmid pFastBac-egfp was constructed, and BmBac-egfp was obtained by transposition, and BmBac-egfp DNA was transfected into BmN (bombyx mori ovary cells ) ...

Embodiment 2

[0037] When the final concentration of small molecules is 0.1 μg / mL, the prevention and treatment of BmNPV infection carrying egfp reporter gene:

[0038] (1) Take 10 of each inoculation 5 For 3 culture dishes of cells, add 2 mL of TC100 insect cell culture medium with 10% FBS to each dish. The old medium was removed, and 0.02 μL of 10 mg / mL small molecule stock solution and 2 mL of 10% FBS TC100 medium were added respectively, incubated at 27°C for 30 min, and then BmBac-egfp virus with MOI=5 was added.

[0039] (2) After 48 hours, place 3 dishes in a fluorescent inverted microscope to observe the expression of green fluorescent protein.

[0040] (3) Take 10 μL of cell culture solution, and measure the virus titer by terminal dilution method.

[0041] For the expression of green fluorescent protein, see figure 1 Middle B, the results show that when cells were treated with 0.1 μg / mL small molecule, the fluorescence of BmNPV-infected cells was significantly reduced compared ...

Embodiment 3

[0043] When the final concentration of small molecules is 0.25 μg / mL, the prevention and treatment of BmNPV infection carrying egfp reporter gene:

[0044] (1) Take 10 of each inoculation 5 For 3 culture dishes of cells, add 2 mL of TC100 insect cell culture medium with 10% FBS to each dish. The old medium was removed, and 0.05 μL of 10 mg / mL small molecule stock solution and 2 mL of 10% FBS TC100 medium were added respectively, incubated at 27°C for 30 min, and then BmBac-egfp virus with MOI=5 was added.

[0045] (2) After 72 hours, place 3 dishes in a fluorescent inverted microscope to observe the expression of green fluorescent protein.

[0046] (3) Take 10 μL of cell culture solution, and measure the virus titer by terminal dilution method.

[0047] For the expression of green fluorescent protein, see figure 1 Middle C, the results show that when the cells were treated with 0.25 μg / mL small molecule, the fluorescence of BmNPV-infected cells was significantly reduced com...

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Abstract

The invention discloses an application of a small molecular compound in the preparation of silkworm antiviral drug. The present invention relates to obtaining small molecular compounds based on mainstream molecular docking techniques, Aiming at the core gene of Bombyx mori baculovirus, the three-dimensional structure of the core gene is obtained by using the virtual modeling technology, the molecular docking screening small molecular database is used for detecting the activity of the obtained small molecular compound, and the small molecular compound with effective effect in the invention is finally obtained. At a final concentration of 1 [mu]g / mL or more, that small molecule compound can completely inhibit replication of BmNPV, and can continue to inhibit replication of BmNPV aft BmNPV completes cell invasion, and has therapeutic effect. The invention can be used for the research and development and production of medicines for preventing and treating blood type pus of silkworm, and has application and popularization value.

Description

technical field [0001] The invention belongs to the field of molecular biology and virology, and relates to the application of a small molecular compound in the preparation of silkworm antiviral infection medicine. Background technique [0002] Bombyx mori blood sputum caused by Bombyx mori Nuclear Polyhedrosis Virus (BmNPV) infection is the most common, harmful and serious infectious disease in silkworm production. At present, there is no specific drug for the treatment of silkworm blood type pus. In production, chemical disinfectants are mainly used to strengthen strict disinfection during silkworm rearing to prevent virus infection and spread. There are mainly chlorine preparations, aldehyde preparations, surfactants and Lime, etc.; or by adding antibiotics to reduce or delay the occurrence of harm, silkworm farmers have an urgent need for blood-type pus disease treatment drugs. Before 1987, it was found that some chemical drugs such as acridine 9-aminolactate, B-propiol...

Claims

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Application Information

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IPC IPC(8): A61K31/4439A61P31/12
CPCA61K31/4439A61P31/12
Inventor 钱平唐旭东卢梦倩张晗俣王永进罗影王宝林
Owner JIANGSU UNIV OF SCI & TECH
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