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Cyclosporin T, its preparation method and application

A technology of cyclosporine and endophytic fungi is applied in the preparation methods of peptides, chemical instruments and methods, fungicides, etc., which can solve the problems of lack of anti-phytopathogenic fungi and other problems, and achieve the effect of excellent disease prevention effect and low dosage.

Active Publication Date: 2018-04-20
LANZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

if possible from Epichloë If we find metabolites that can inhibit plant pathogenic fungi, and these metabolites are non-toxic to livestock, then inoculating these endophytic fungi in common pastures to make them resistant good pastures will solve the problem of the lack of resistant plants in my country. The problem of good forage grass for pathogenic fungi

Method used

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  • Cyclosporin T, its preparation method and application
  • Cyclosporin T, its preparation method and application
  • Cyclosporin T, its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Preparation of Cyclosporin T:

[0031] 1. Experimental method:

[0032] will first Epichloë bromicola Endophytic fungi were inoculated in potato dextrose liquid medium and cultured on a shaker in a constant temperature box at 27°C for 7 days, and then inoculated into 100 1000 mL Erlenmeyer flasks, each containing 100 g of wet The rice was cultivated for 30 days in an incubator at 27°C. At this point the rice medium was extracted with ethyl acetate to obtain 18 g of crude extract, which was then chromatographed on a silica gel column (chloroform-methanol as solvent) and Sephadex to obtain Cyclosporin T (478 mg). The chemical structure of Cyclosporin T was determined by NMR, high-resolution mass spectrometry and comparison with literature.

[0033] 2. Experimental results:

[0034] right Epichloë bromicola Fermented under rice culture medium, a large amount of Cyclosporin T can be obtained.

experiment example 2

[0036] Observation of Cyclosporin T on normal cytotoxicity of bovine kidney:

[0037] 1. Experimental method:

[0038] Normal bovine kidney cells were inoculated in a 96-well plate, and 100 μL of DMEM medium was added to each well, and cultured at 37°C for 24 hours. Cyclosporin T with different concentration gradients was added to the 96-well plate, and cultured for another 24 hours. Afterwards, 10 μL of 5 mg / mL resazurin sodium solution was added to the wells, and after incubation for 2 hours, the fluorescence intensity was measured at a wavelength of 555 / 585 nanometers with a multifunctional enzyme labeling instrument to calculate the inhibition rate.

[0039] 2. Experimental results:

[0040] IC of Cyclosporin T on bovine kidney cells 50 >100 μM, and from 12.5 μM to promote the growth of bovine kidney cells, indicating that Cyclosporin T is non-toxic to livestock, Epichloë bromicola is a beneficial endophytic fungus.

Embodiment 3

[0042] Antibacterial activity of Cyclosporin T against mycelial growth of phytopathogenic fungi:

[0043]1. Pathogenic fungi for testing: Alternaria ( Alternaria alternat ), Isohelmosporium tritici ( Bipolaris sorokinian ), Fusarium avenatum ( Fusarium avenaceum ), Curvularia lunata ( Curvularia lunata ).

[0044] 2. Experimental method:

[0045] The inhibitory effect of rhododendron on the growth of phytopathogenic fungal mycelia was determined by the growth rate method in a toxic medium. The brief description is that Cyclosporin T is dissolved in DMSO and then added to molten PDA medium (DMSO content 5 ‰), respectively formulated into drug-containing PDA plates containing a certain concentration of Cyclosporin T. Move the hyphae blocks of each pathogenic fungus with a diameter of 5 mm after cultivation and activation to the center of the drug-containing plate, set three repetitions for each concentration, and set 5‰ of DMSO as a control. Set 27 ℃ constant tempera...

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Abstract

The invention discloses cyclosporine T, the structural formula of which is shown in formula (I), and provides its preparation method and application. The cyclosporine T provided by the present invention and its preparation method and application have the beneficial effects as follows: (1) cyclosporin T is non-toxic to livestock, and its source Epichloë bromicola is a beneficial endophytic fungus; (2) Cyclosporin T has broad-spectrum bactericidal activity, low dosage, and better disease prevention effect than positive drugs; (3) Cyclosporin T is a natural product, which can be obtained by mass fermentation of Epichloë bromicola endophytic fungi, and is sustainable (4) Epichloë bromicola endophytic fungus can be inoculated on other disadvantaged forages, and it has the potential to cultivate dominant forages resistant to phytopathogenic fungi.

Description

technical field [0001] The invention relates to the field of compound preparation and application, in particular to cyclosporine T and its preparation method and application. Background technique [0002] The slow development of grassland agriculture in my country has the most direct impact on the slowdown in the development of animal husbandry, which has led to an increase in the amount of pasture grass imported from abroad and the purchase of infant milk products in my country every year. The main reason is that my country lacks excellent forage species with resistance, especially forage species with resistance to phytopathogenic fungi. Every year, due to the harm of plant pathogenic fungi, pastures in our country cause large-scale production reduction and slow growth, and also affect the safety of dairy products. Although chemically synthesized drugs for killing phytopathogenic fungi at the present stage play an irreplaceable role in plant disease control. However, peop...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/64C07K1/16G01N33/68A01P3/00
Inventor 宋秋艳高坤南志标田沛宋辉
Owner LANZHOU UNIVERSITY
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