A kind of application of insect cell immunosuppressant

An immunosuppressant and insect cell technology, which is applied in the field of natural microorganism-derived active product compounds, can solve the problems of no insecticidal activity of the second instar armyworm and no significant impact on the immune system, and achieves enhanced infectious death and good pest control. and pest physiological adaptive regulation, the effect of good application prospects

Active Publication Date: 2020-04-03
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chinese patent document CN101684448A discloses the preparation method of the compound and the insecticidal activity to the second instar armyworm, but it was found that the compound had no insecticidal activity to the second instar armyworm when the concentration was lower than 3.125ppm (=3.125 μg / mL)
Among the high-efficiency, low-toxicity and environment-friendly insecticides commercialized at home and abroad such as: tebufenozide (tebufenozide), imidacloprid (imidacloprid), etc., although they all have superior insecticidal activity, they are isolated from insects. Neither somatic nor insect cellular immune systems were significantly affected

Method used

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  • A kind of application of insect cell immunosuppressant
  • A kind of application of insect cell immunosuppressant
  • A kind of application of insect cell immunosuppressant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Spiral ketal compound 5'-epi-SPA-6952A inhibits insect cell viability

[0030] Experimental Materials:

[0031]Diptera Drosophila melanogaster (Drosophila melanogaster) Schneider's S2 cells were purchased from the Institute of Cells, Chinese Academy of Sciences. The spiro ketal compound 5'-epi-SPA-6952A was prepared by referring to the method disclosed in the patent document CN101684448A. TNM-FH insect cell culture medium was purchased from HyClone Company. Trypsin and penicillin (100UI) / streptomycin (100mg / mL) mixed antibiotics were purchased from Gibico. Thiazolium blue (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, MTT) was purchased from Sigma.

[0032] experimental method:

[0033] (1) Take Tn-5B1-4 cells in the logarithmic growth phase and adjust the cell density to 5×10 4 Cells / mL were seeded in 96-well culture plates until the cell monolayer grew to 80% confluence.

[0034] (2) Add 100 μL of the test compound solution to the expe...

Embodiment 2

[0041] Example 2 Spiral ketal compound 5'-epi-SPA-6952A induces apoptosis in insect cells

[0042] Experimental Materials:

[0043] Lepidoptera insect Trichoplusia ni Tn-5B1-4 cells were purchased from the Institute of Cells, Chinese Academy of Sciences. Annexin V-FITC Apoptosis Detection Kit and propidium iodide (PI) were purchased from Shanghai Beyotime Institute of Biotechnology. FACS Calibur flow cytometer is a product of Becton Dickinson Company of the United States.

[0044] experimental method:

[0045] (1) Inoculate 1×10 5 cell / mL Tn-5B1-4 cells in the logarithmic growth phase were placed in a 6-well plate until the cell monolayer reached 80% confluence.

[0046] (2) In the experimental group, 4 mL of the test compound solution was added to the final concentrations of 0.008, 0.016, and 0.032 μg / mL. In the control group, 4 mL of TNM-FH cell culture medium was added. Set more than 3 parallel wells in each group, place at 37°C, 5% CO 2 and saturated humidity incuba...

Embodiment 3

[0053] Example 3 Spiro ketal compound 5'-epi-SPA-6952A induces morphological changes in insect hemocytes

[0054] Experimental Materials:

[0055] TNM-FH insect cell culture medium was purchased from HyClone Company. Giemsa stain was purchased from Sigma.

[0056] experimental method:

[0057] (1) Collection and in vitro culture of blood cells. After the body surface of the 5th instar Spodoptera litura larvae was washed with distilled water, it was disinfected with 75% alcohol. The gastropod was punctured with a sterile needle, and the hemolymph was collected in an Eppendorf centrifuge tube containing a small amount of phenylthiourea crystals, mixed gently, and kept in an ice bath for later use.

[0058] (2) In vitro culture of blood cells. Take 20 μL of hemolymph in a 24-well cell culture plate, add TNM-FH insect medium, and then place it in a constant temperature incubator at 28°C for cultivation.

[0059] (3) In the experimental group, 20 μL of the test compound solut...

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PUM

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Abstract

The invention relates to the field of application of spiroketal compounds, in particular to application of a compound, namely 5'-epi-SPA-6952A, to preparation of insect immunosuppressor. The compound, namely 5'-epi-SPA-6952A, restrains the functions of an insect immune system, and the structural formula of the compound, namely 5'-epi-SPA-6952A, is shown in the description. The compound has a novel action mechanism, the restraining effect of the compound on the injurious insect immune system is different from that of action mechanisms of existing insecticides, and the compound is not influenced by the drug resistance of insects to other insecticides, has the advantage of being friendly to the environment, and has good application prospects on insect control and injurious insect physiological adaptation regulation and control.

Description

technical field [0001] The present invention relates to the technical field of use of spiro ketal compounds, in particular, the natural microbial source active product compound 5'-epi-SPA-6952A is used to inhibit insect cell immunity. Background technique [0002] Insect damage is an important limiting factor of the world's food production. The loss caused by insect pests in the world accounts for about 15% of the total crop harvest, and the annual loss is nearly 100 billion U.S. dollars. Chemical control is the most effective measure for plant protection in modern agriculture, and it plays an irreplaceable and important role in the control of pest damage on food crops. People use pesticides to control crop pests, which improves the yield of crops and ensures a good harvest of crops. However, due to long-term dependence on chemical pesticides, extensive use of chemical pesticides, or irrational use of pesticides, not only lead to insect resistance, make pest control more co...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N43/90A01P7/04A01P15/00A01G13/00
CPCA01G13/00A01N43/90
Inventor 黄青春肖慈英朱琦琦张蕾杨超陶黎明郓新明饶文兵
Owner EAST CHINA UNIV OF SCI & TECH
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