Antitumor antibiotic gaddie mycin generated by streptosporangium C-3560

A technology of gadomycin and streptocyst, applied in the field of biomedicine

Active Publication Date: 2014-05-07
MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Streptosporangium C-3560 provided by the present invention produces the antitumor antibiotic gadomycin and its application, so far, there are no relevant reports at home and abroad

Method used

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  • Antitumor antibiotic gaddie mycin generated by streptosporangium C-3560
  • Antitumor antibiotic gaddie mycin generated by streptosporangium C-3560
  • Antitumor antibiotic gaddie mycin generated by streptosporangium C-3560

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] "Example 1" culture characteristics of streptocystis C-3560

[0039] Morphological characteristics: After 15 days of culture on glucose asparagine agar and oat flour agar inserts, the C-3560 strain formed round, oval or irregular cysts with a length of about 8-12 μm. The spores in the cyst are arranged in a single circle, elliptic, without flagella, immobile, and have a smooth surface. The mycelium in the base has no diaphragm and does not break. The aerial hyphae are milky white to white, medium in length, and the branched spores are short, straight and smooth.

[0040] Shape characteristics: The culture characteristics of the C-3560 strain on 8 kinds of media are shown in Table 2.

[0041] The culture characteristics of table 2C-3560 bacterial strain

[0042]

[0043] 16S rDNA sequence analysis of Streptocystis sp. C-3560:

[0044] The comparison of the 16S rDNA sequence of strain C-3560 with GenBank showed that the strain belonged to the genus Streptosporangiu...

Embodiment 2

[0045] "Example 2" fermentation broth preparation of Streptocystis sp. C-3560

[0046] Streptocystis C-3560 strain preserved in glycerol tubes was inoculated on the slant medium. Slant medium by weight: yeast extract 0.4, malt extract 1.0, glucose 1.0, oat flour 1.0, FeSO 4 ·7H 2 O 0.0001, MnCl 2 4H 2 O 0.0001, ZnSO 4 ·7H 2O 0.0001, agar 1.4-1.8, distilled water 100, pH value 7.0. Constant temperature and static culture at 28-30°C for 10-14 days. Inoculate the slant culture into 1000ml seed medium (the weight ratio of the seed medium is: soluble starch 1.0, maltose 2.0, NaCl 0.3, digested serum powder 0.5, yeast extract 0.2, soybean powder 1.0, FeSO 4 ·7H 2 O 0.0001, MnCl 2 4H 2 O 0.0001, ZnSO 4 ·7H 2 O 0.0001, distilled water 100). 28-30°C constant temperature, 190rpm shaking culture for 72 hours. Take the cultivated seed culture solution and add it into 10L fermentation medium at a ratio of 10% (the fermentation medium is: dextrin 1.0, maltose 2.0, NaCl 0.3, dig...

Embodiment 3

[0047] 《Example 3》Multi-step chromatography prepares gamidomycin

[0048] The fermentation broth of the C-3560 strain was centrifuged at 5000rpm at 4°C for 20min at low temperature, and the supernatant was taken. The supernatant was concentrated by a 3kDa ultrafiltration membrane module (CLW-003 type, molecular weight cut-off 3kDa, Beijing Xubang Membrane Equipment Co., Ltd.), and 1L of crude concentrate was obtained for the following multiple chromatography:

[0049] 1) DEAE-cellulose weak anion exchange column chromatography

[0050] 1L of the crude concentrate was centrifuged at 5000rpm at 4°C for 20min at low temperature to obtain the supernatant; it was subjected to DEAE-cellulose (Whatman DE52 type) weak anion exchange column chromatography (Na + type, pre-equilibrated with deionized water), the column volume of the exchange column is 180ml, and the column diameter ratio is 9:48 to collect the flow-through. Elute with 500ml 0.05mol / l NaCl solution. Collect the eluate ...

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Abstract

The invention relates to an antitumor active substance generated by streptosporangium C-3560. The antitumor active protein molecular weight is 10.8kDa, and the N-end sequence is shown as a sequence table SEQ ID No.1. Active substance biology experimental study results prove that obvious antitumor activity is realized on various kinds of tumor cells in-vitro culture, the tumor cell apoptosis can be induced, and in addition, the cell cycle arrest is generated. In-vitro animal experiments prove that the tumor growth of mouse H22 liver cancer can be obviously inhibited, and the antitumor active substance is hopeful to further develope into clinical efficient antitumor medicine.

Description

Technical field: [0001] The invention belongs to the field of biomedicine and relates to an antitumor active substance produced by Streptocystis sp., in particular to a protein with antitumor activity and an N-terminal sequence and its application in the preparation of antitumor drugs . Background technique: [0002] So far, more than 10 kinds of antitumor antibiotics derived from microorganisms have been used clinically as anticancer drugs. They all originate from the secondary metabolites of microbial fermentation or the semi-synthesis of microbial secondary metabolites. Due to the diversity of sources of microorganisms and the diversity of secondary metabolites in nature, a large number of antitumor active substances produced by microorganisms have been reported. However, there are still many unsatisfactory aspects of the pharmaceutical properties of many known antitumor antibiotics used clinically (such as toxicity, efficacy, indications, and production of drug-resista...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/195C12P21/02A61K38/16A61P35/00C12R1/62
Inventor 王莹甄永苏胡继兰李毅范翠敏尚柏杨
Owner MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
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