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Emiclamycin and its preparation method and application

A technology of esomeclamycin and pharmacy, applied in the field of biopharmaceuticals, can solve the problems of narrow dose range, poor tumor specificity, poor penetration, etc., and achieve the effect of overcoming the shortcomings of antibiotic resistance and strong cytotoxic activity

Active Publication Date: 2022-03-25
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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  • Emiclamycin and its preparation method and application
  • Emiclamycin and its preparation method and application
  • Emiclamycin and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0046] The Asian locusts (Locusta migratoria) were stocked under starvation conditions for 24 hours, and then sterilized their shells with 75% alcohol in an ultra-clean bench under aseptic conditions, and then removed the locust intestines and soaked them in 75% alcohol for 2 minutes for surface treatment. Sterilize, then rinse with sterile water 3 times, add a small amount of sterile water to grind in a sterile mortar, and use sterile water to dilute the grinding liquid into 3 gradients of 10 -1 , 10 -2 , 10 -3 , respectively, take 200 μl of each gradient dilution and spread it on Gao’s No. 1 medium (soluble starch 20 g, 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 (0.01g, agar 15-25g, distilled water 1L, PH7.4~7.6), place in 28 ℃ of incubators and cultivate, wait for the colony to grow (after about ten days), pick a single colony with a bamboo s...

Embodiment 2

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

[0048] Activated Amycolatopsis sp.HCa4, inoculated fresh mycelium blocks into 1L shake flasks, each flask contained 200mL of TSB medium (tryptone 15g, soy peptone 5g, sodium chloride 5g, distilled water 1L), placed on a shaker Cultivated at 28°C for 2 days and used as seed solution. The inoculation volume of 30 mL per bottle was inoculated into 100 flasks containing 300 mL of I culture solution (5 g of peptone, 3 g of yeast powder, 10 g of starch, 1 L of water), and cultured at 28° C. and 230 rpm / min for 8 days.

Embodiment 3

[0049] Example 3: Extraction and isolation of Amycolamycin

[0050] The fermentation broth obtained in Example 2 was filtered with 4 layers of gauze, the filtrate was extracted 3 times with an equal amount of ethyl acetate, and the red crude extract F1 was obtained by vacuum concentration and drying. The crude extract F1 was suspended in water, extracted with petroleum ether and ethyl acetate in turn, and the obtained ethyl acetate extract was concentrated to obtain extract F25.1g; the extract F2 was subjected to normal phase silica gel column chromatography, and CH 2 Cl 2 / MeOH as mobile phase gradient elution (v / v 100:1, 100:2, 100:4, 100:8, 100:16), 4 column volumes, divided into 15 fractions. According to the results of LC-MS analysis, after the third component was separated and purified by Sephadex LH-20, the flow rate of 2.0mL / min was carried out by semi-HPLC (chromatographic column: AllsphereODS-2.5mm column) under the condition of acetonitrile water 20-50%. Gradient ...

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Abstract

The invention discloses an emaclamycin and its preparation method and application. The said emaclamycin is derived from the liquid fermentation product of the endophytic actinomycete Amycolatopsis sp.HCa4 in the intestinal tract of migratory locust, including emaclamycin A or emaclamycin B. Experimental results show that emaclamycin has strong antibacterial activity against methicillin-resistant Staphylococcus aureus and strong cytotoxic activity against human breast cancer cell M231, and can be prepared into a new type of anti-tumor lead compound, to a certain extent Overcome the shortcomings of existing antibiotic resistance.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals, and in particular relates to an emeklemycin, a preparation method and application thereof. Background technique [0002] Insect guts are inhabited by a large number of microorganisms, and the early research on insect gut microbes mostly focused on the classification and application of insect microorganisms. 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 mutually beneficial 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 metabol...

Claims

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Application Information

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Patent Type & Authority Patents(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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