Alternarlactam and preparation method and application thereof

A technology of alternaria and alkaloids, applied in the field of microbial engineering and pharmaceutical engineering, to achieve the effects of overcoming drug resistance defects, low cost, and simple process

Inactive Publication Date: 2013-04-17
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Many chemically synthesized anticancer drugs have very strong side effects on normal cells while killing tumor cells, and tumor cells are prone to multidrug resistance during treatment, which often leads to treatment failure.

Method used

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  • Alternarlactam and preparation method and application thereof
  • Alternarlactam and preparation method and application thereof
  • Alternarlactam and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1: Isolation and purification of fungi of the genus Alternaria sp. from leaves of Carex aridula V. Krecz.

[0040] Cut the sedge leaves, wash them with tap water, then cut the leaves into small pieces of 1cm×1cm, then disinfect with 75% ethanol for 1min, use 2.5% sodium hypochlorite for 10min, then sterilize with 75% ethanol for 1min, and then use no Rinse with bacterial water 2-3 times, insert on the plate of WA+double antibody medium, and culture at 28°C for 3-15 days. After the thalli grew out, the mycelium at the edge of the colony was picked for purification and culture to obtain the Carex leaf fungus. Colony dark green to black, hyphae pale, growing rapidly; conidiophores dark, single, variable in length, terminal unbranched or occasionally branched spore chains; conidia dark, with longitudinal and transverse septa, inverted clubs Shaped, oval or ovate, often forming chains with beak-like appendages at the top. It was identified as Alternaria sp.HG1 by A...

Embodiment 2

[0041] Embodiment 2: the solid fermentation of Carex blade Alternaria sp. (Alternaria sp.) fungus

[0042] The mycelium of Alternaria sp. activated on the PDA plate medium was inoculated in 20 bottles of PD medium (300mL per bottle), and in a rotary shaker (28°C, 150rpm) Shaking culture was carried out for 6 days, and the fermented liquid was inoculated in 400 bottles of millet solid medium, with 15 mL of bacterial liquid in each bottle. The inoculated solid medium was cultured statically at 28°C for 30 days.

Embodiment 3

[0043] Embodiment 3: Extraction and separation of Alternaria alkaloid (alternarlactam)

[0044] The solid culture obtained in Example 2 was dried, ground and weighed to obtain 2000 g of solid. The solid culture was extracted five times with 3 L of chloroform / methanol mixed solvent (1:1, V / V), and the extract was concentrated to obtain crude extract F1. Dissolve the crude extract F1 with an appropriate amount of methanol, then place the solution in a refrigerator at -4°C overnight, filter to remove the precipitated waxy solid, and concentrate the filtrate under reduced pressure to obtain extract F2. The extract F2 was subjected to silica gel column chromatography, and first eluted with 10 times the volume of chloroform to obtain the extract F3. Then continue to elute the extract F2 with chloroform / methanol (volume ratio 100 / 1) mixed solution elution, and combine the eluents of the white parts to obtain the white extract F4. For the extract F4, the L-alternaria alkaloid [(-)-a...

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Abstract

The invention belongs to the technical field of microbial engineering and pharmaceutical engineering and particularly relates to alternarlactam extracted from a solid fermentation material of maubertiana boott endophytic fungi (Alternaria sp.) and a preparation method and anti-tumor effect thereof. The alternarlactam can be prepared by a solid fermentation method and a chemical synthesis method. The alternarlactam has the effects of strongly inhibiting the proliferation of human cervical cancer cells HeLa and human liver cancer cells QSG-7701, so that the alternarlactam can be used as a compound having the anti-tumor effect and is applied to preparing anti-tumor medicaments.

Description

technical field [0001] The invention belongs to the technical field of microbial engineering and pharmaceutical engineering, and in particular relates to a class of Alternaria alkaloids extracted from the solid fermented product of Carex dense leafy fungus (Alternaria sp.), a preparation method thereof and antitumor effect. Background technique [0002] Plant endophytes refer to those fungi and bacteria (including actinomycetes) that live in various tissues of healthy plants during certain or all stages of their life history and do not cause obvious infection symptoms of the host. There is a mutualistic symbiotic relationship between plant endophytes and hosts, which can promote the growth of host plants, improve the host's ability to resist adversity, pathogens, and animal hazards, and make host plants more viable than uninfected plants. Competitiveness. Due to their unique habitat, endophytes can produce a variety of secondary metabolites. They often have novel structure...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12P17/12C07D221/16A61K31/473A61P35/00C12R1/645
Inventor 谭仁祥张爱华谷文宋勇春王玉蓉胥苏海
Owner NANJING UNIV
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