Application of small molecule compound in preparation of drugs for resisting baculovirus infection of silkworms

A technology of small molecular compounds and silkworm baculovirus, which can be used in antiviral agents, medical preparations containing active ingredients, pharmaceutical formulations, etc., can solve the problem that the development of silkworm drugs and silkworm antiviral drugs has not kept up with the development of silkworm drugs and has little capital investment. , Weak R&D strength, etc.

Active Publication Date: 2019-02-22
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

Method used

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  • Application of small molecule compound in preparation of drugs for resisting baculovirus infection of silkworms
  • Application of small molecule compound in preparation of drugs for resisting baculovirus infection of silkworms
  • Application of small molecule compound in preparation of drugs for resisting baculovirus infection of silkworms

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] 1. Construction of BmNPV carrying green fluorescent protein reporter gene

[0022] 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.

[0023] The specific method of this step is as follows:

[0024] 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

[0036] When the final concentration of small molecules is 0.1 μg / mL, the prevention and treatment of BmNPV infection carrying egfp reporter gene:

[0037] (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.

[0038] (2) After 48 hours, place 3 dishes in a fluorescent inverted microscope to observe the expression of green fluorescent protein.

[0039] (3) Take 10 μL of cell culture solution, and measure the virus titer by terminal dilution method.

[0040] 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

[0042] When the final concentration of small molecules is 0.25 μg / mL, the prevention and treatment of BmNPV infection carrying egfp reporter gene:

[0043] (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.

[0044] (2) After 72 hours, place 3 dishes in a fluorescent inverted microscope to observe the expression of green fluorescent protein.

[0045] (3) Take 10 μL of cell culture solution, and measure the virus titer by terminal dilution method.

[0046] 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 application of a small molecule compound in preparation of drugs for resisting baculovirus infection of silkworms. The involved small molecule compound is obtained on the basisof a mainstream molecular docking technology, for a necessary core gene of the baculovirus infection of the silkworms, a three-dimensional structure of the core gene is obtained by a virtual modelingtechnology, a small molecule database is screened by molecular docking, the obtained small molecule compound is subjected to activity detection, and finally, the small molecule compound with effectsin the invention is obtained finally. When the using fine concentraton of the small molecule compound is 1 mu g/mL or above, copying of BmNPV can be inhibited completely, after cell infection by BmNPV, copying of BmNPV can be still inhibited, and a therapy effect is achieved. The small molecule compound can be used for researching and producing drugs for preventing and treating nuclear polyhedrosis of the silkworms, and the small molecule compound has application and promotion 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 medicine against silkworm infected by baculovirus. 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-am...

Claims

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Application Information

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IPC IPC(8): A61K31/165A61P31/20
CPCA61K31/165A61P31/20
Inventor 唐旭东张晗俣吕丁丁蒋师东张轶岭
Owner JIANGSU UNIV OF SCI & TECH
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