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Amycolamycin as well as preparation method thereof and application thereof

A kind of Emmycladamycin, a pharmaceutical technology, applied in the field of biopharmaceuticals, can solve the problems of narrow dose range, poor tumor specificity, poor penetration, etc., and achieve the effects of low environmental pressure, simple preparation process and low cost

Active Publication Date: 2019-04-26
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, clinical deficiencies such as narrow dose range and poor tumor specificity are mainly solved by using monoclonal antibodies as carriers to enhance targeting, and using specific "warhead" drugs linked to different monoclonal antibodies to solve human rejection and penetration Poor and other issues

Method used

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  • Amycolamycin as well as preparation method thereof and application thereof
  • Amycolamycin as well as preparation method thereof and application thereof
  • Amycolamycin as well as preparation method thereof and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Isolation, purification and identification of Amycolatopsis sp.HCa4

[0046] Put Asian migratory locusts (Locusta migratoria) in a suitable place for 24 hours under starvation conditions, and then disinfect their shells with 75% alcohol in an ultra-clean bench under aseptic conditions, then take out the locust intestines and soak them in 75% alcohol for 2 minutes for surface treatment. Disinfect, then rinse 3 times with sterile water, add a small amount of sterile water to grind in a sterile mortar, and use sterile water to gradiently dilute the grinding liquid into 3 gradients of 10 -1 , 10 -2 , 10 -3 , take 200 μl of each gradient dilution solution and spread it on Gao’s No. 1 medium (soluble starch 20g, KNO 3 1g, NaCl 0.5g, K 2 HPO 4 ·3H 2 O 0.5g, MgSO 4 ·7H 2 O 0.5g, FeSO 4 ·7H 2 O 0.01g, agar 15-25g, distilled water 1L, pH7.4~7.6), cultured in a 28°C incubator, and when the colony grows (about ten days later), pick a single colony with a bamboo...

Embodiment 2

[0047] Example 2: Liquid fermentation of Amycolatopsis sp.HCa4

[0048] To activate Amycolatopsis sp.HCa4, inoculate fresh mycelium blocks into 1L shake flasks, each containing 200mL of TSB medium (tryptone 15g, soybean peptone 5g, sodium chloride 5g, distilled water 1L) and place on a shaker Cultivate at 28°C for 2 days and serve as seed solution. Inoculate 100 bottles of shake flasks containing 300 mL of I culture solution (5 g of peptone, 3 g of yeast powder, 10 g of starch, and 1 L of water) with an inoculum size of 30 mL per bottle, and cultivate for 8 days at 28° C. and 230 rpm / min.

Embodiment 3

[0049] Embodiment 3: Extraction and separation of Amycolamycin

[0050] The fermented liquid obtained in Example 2 was filtered with 4 layers of gauze, the filtrate was extracted 3 times with an equal amount of ethyl acetate, concentrated and dried in vacuo to obtain red crude extract F1. The crude extract F1 was suspended in water, extracted with petroleum ether and ethyl acetate successively, and the obtained ethyl acetate extract was concentrated to obtain 25.1 g of extract F2; the extract F2 was subjected to normal phase silica gel column chromatography, and CH 2 Cl 2 / MeOH as mobile phase gradient (v / v 100:1, 100:2, 100:4, 100:8, 100:16), 4 column volumes, divided into 15 fractions. According to the LC-MS analysis results, after the third component was separated and purified by Sephadex LH-20, it was separated and purified by semi-HPLC (chromatographic column: AllsphereODS-2.5mm column) under the condition of 20-50% acetonitrile water at a flow rate of 2.0mL / min Amycola...

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Abstract

The invention discloses amycolamycin as well as a preparation method thereof and application thereof. The amycolamycin is from a liquid fermentation product of endophytic actinomycetes Amycolatopsis sp.HCa4 in the intestinal tract of asiatic migratory locust, and comprises amycolamycin A or amycolamycin B. According to the experimental research results, the amycolamycin has very strong antibacterial activity on methicillin-resistant staphylococcus epidermidis staphylococcus aureus, has very strong cytotoxic activity on human breast cancer cells M231, can be prepared into a novel anti-tumour lead compound, and overcomes the drug resistance defect of existing antibiotics to a certain extent.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals, and in particular relates to an emaclamycin, its preparation method and application. Background technique [0002] Insect guts inhabit a large number of microorganisms. Early studies on insect gut microbes mostly focused on the classification and application of insect microbes. With the deepening of research, some researchers have found that there is a wide range of non-pathogenic relationships between symbiotic bacteria and insect hosts, including mutualistic symbiotic relationships, and even some microorganisms are necessary for the growth, development and reproduction of the host. Microorganisms in this special symbiotic relationship are likely to produce some new compounds with novel structures and better activities during their metabolism. [0003] Antitumor antibiotics refer to substances with antitumor activity produced by microbial metabolism, which can be divided into: enediy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H17/08C12P17/18A61K31/7048A61P35/00A61P31/04C12R1/01
CPCC12P17/181C07H17/08
Inventor 戈惠明谭仁祥马仕英肖永胜张博郭志凯
Owner NANJING UNIV
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