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Marine natural anti-glioma active substance, and preparation and application thereof

A technology of bacteria and culture, applied in the preparation of anti-glioma drugs, the field of marine natural anti-glioma active substances

Active Publication Date: 2016-06-15
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, unfortunately, there are no anti-glioma drugs related to natural products in clinical use in the world at present.

Method used

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  • Marine natural anti-glioma active substance, and preparation and application thereof
  • Marine natural anti-glioma active substance, and preparation and application thereof
  • Marine natural anti-glioma active substance, and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1. Isolation and culture of Streptomyces filipinensis ZQ-22

[0036] Take 1 gram of air-dried mangrove soil and dilute it with seawater to a concentration of 1×10 -6 g / mL sample diluent, take 200μL of sample diluent and evenly disperse it into a petri dish containing Gauss's agar (Gause'sagar) solid medium. After culturing at 28℃ for 7 days, transfer the different colonies to In another petri dish containing Gao's agar solid medium, the culture was continued for 7 days at 28°C. Finally, the well-growing single colony (ZQ-22) was inoculated into the Gauss' agar slant medium for cultivation and then stored in the refrigerator at 4°C for later use.

Embodiment 2

[0037] Example 2. Identification of Streptomyces filipinensis ZQ-22

[0038] The 16SrDNA sequence analysis method was used to identify the type of the obtained strain ZQ-22.

[0039] 2.1 Experimental reagents and instruments

[0040] PCR reagents: EXTaq enzyme (TaKaRa), dNTP (TaKaRa), primers (synthesized by Invitrogen), the primer sequence is: TACGGYTACCTTGTTACGACTT and AGAGTTTGATCMTGGCTCAG;

[0041] Marker: DL5000;

[0042] Experimental instruments: centrifuge, electrophoresis machine, PCR machine, ABI3730XL sequencer.

[0043] 2.2 Experimental steps

[0044] Bacterial genomic DNA extraction

[0045] Electrophoresis detection

[0046] PCR amplification

[0047] a. PCR reaction system

[0048]

[0049] b. PCR reaction conditions

[0050]

[0051] c. Electrophoresis detection

[0052] d. Sequencing: gel cutting, purification and sequencing

[0053] e. Analysis result: splicing sequence.

[0054] 2.3 Experimental results

[0055] The spliced ​​sequence is:

[0056] GGGCCACCGGCTTCGGGTGTTACCGACTTTCGTG...

Embodiment 3

[0058] Example 3. Preparation of fermentation broth of Streptomyces filipinensis ZQ-22

[0059] Streptomyces filipinensis ZQ-22 was inoculated into a 500 mL Erlenmeyer flask containing 200 mL of liquid Gaussian medium, and the culture solution containing ZQ-22 strain was cultured at 28°C with rotation (180 rpm) for 7 days. Seed liquid. Transfer 5 mL of the bacterial seed liquid into a 500 mL Erlenmeyer flask containing 200 mL of liquid Gaussian medium, and rotate (180 rpm) at 28° C. and shake culture for 7 days to obtain a ZQ-22 fermentation broth with anti-tumor activity.

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Abstract

The invention relates to a marine natural anti-glioma active substance, and preparation and application thereof. The invention provides a Streptomyces filipinensis strain of which the classification designation is Streptomyces filipinensis ZQ-22. The Streptomyces filipinensis ZQ-22 is separated and cultured from soil under mangrove. The Streptomyces filipinensis is subjected to fermentation and extraction separation purification to obtain the anti-glioma active compound Bagremycin C. The compound obviously inhibits different glioma cells from proliferation, obviously induces the tumor cell apoptosis and influences the tumor cell period, and can be applied to preparation of drugs for treating glioma. The chemical structural formula of the Bagremycin C is disclosed in the specification.

Description

Technical field [0001] The invention belongs to the technical field of marine biology, and relates to a method for preparing a new marine natural anti-glioma active substance Bagremycin C from the marine actinomycete Streptomyces filipinensis (Streptomyces filipinensis ZQ-22) and the compound in the preparation of anti-glioma drugs Applications. Background technique [0002] Gliomas are the most common brain tumors in the central nervous system with extremely high mortality, accounting for about 70% of all malignant brain tumors. A combination of radiotherapy and chemotherapy after surgery is currently the most commonly used treatment for glioma. Because gliomas involve many important brain functional areas, it is extremely difficult to remove gliomas. Therefore, compared with other tumors, the treatment of gliomas with drugs is more important. However, there is a severe shortage of drugs used to treat gliomas in the world, and there are only a few including temozolomide (TMZ),...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02C12P13/02C07C319/28C07C323/63A61K31/245A61P35/00C12R1/53
CPCC07C319/28C07C323/63C12N1/02C12P13/02C12N1/205C12R2001/53
Inventor 张治针陈磊连晓媛
Owner ZHEJIANG UNIV
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