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Yuanjiang mycin analogue as well as preparation method and application thereof

A technology of Yuanjiangmycin and analogues, applied in biochemical equipment and methods, microorganism-based methods, microorganisms, etc., can solve problems that hinder the discovery of new compounds

Pending Publication Date: 2021-06-08
HAYAO CIHANG PHARM CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, under laboratory conditions, most clusters are often not expressed or expressed very little, hindering the discovery of new compounds

Method used

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  • Yuanjiang mycin analogue as well as preparation method and application thereof
  • Yuanjiang mycin analogue as well as preparation method and application thereof
  • Yuanjiang mycin analogue as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Preparation and structure analysis of YJM-A and YJM-B

[0039] A 1 mL spore culture of strain Streptomyces sp. CB03234-S was grown in a 2000 mL Erlenmeyer flask containing 500 mL tryptic soy broth (TSB) for 36 hours. The seed culture (10vol%) was transferred to the fermentation medium (soluble starch 10g / L, cottonseed meal 5g / L, CuSO 4 0.05g / L, NaI 5mg / L, CaCO 3 2g / L, in 1% macroporous resin HP20) and cultivated for 4 days at 30°C and 220rpm, and the fermentation volume was 20L.

[0040] The resin was separated by centrifugation or filtered through a metal sieve (60 mesh), washed with pure water and air dried at room temperature. It was then eluted three times each with 3 L of EtOH and concentrated under vacuum. In 2L EtOAc:H 2O (1:1) was extracted three times to obtain the ethyl acetate soluble fraction. The ethyl acetate extract was concentrated in vacuo to give a crude extract. The crude extract (32.5g) was mixed with 80-100 mesh silica gel, packed into a col...

Embodiment 2

[0062] In vitro anti-tumor activity test:

[0063] The experimental method is as follows:

[0064] In this experiment, the cytotoxicity of YJMA and YJMB was tested by CCK-8 colorimetric method. The cancer cell density was adjusted to 1×104 per well, inoculated in a 96-well plate, and incubated in a 5% CO2 incubator at 37°C for 24 hours. Dilute YJM A and YJM B in 1.5 mL EP tubes with DMEM medium in 7 concentration gradients, 0.002 μM, 0.018 μM, 0.182 μM, 1.818 μM, 9.091 μM, 18.182 μM, 45.455 μM and 90.909 μM, respectively. Place in an incubator and incubate for 72h. After adding the CCK-8 solution, incubate for 2-4 hours according to the specific situation, measure the absorbance value with a microplate reader, perform statistical analysis on the data and draw a graph. The cell survival rate is calculated as shown in the formula:

[0065]

[0066] The results are shown in Table 3.

[0067] Table 3: Cytotoxicity of YJM A and YJM B against various tumor cell lines

[006...

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Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to a Yuanjiang mycin confirmation method, a preparation method and an anti-tumor application of Yuanjiang mycin. The structural formula of the Yuanjiang mycin analogue is shown in the specification. According to the invention, through fermentation culture of Streptomyces sp.CB03234-S, then design of special separation and purification steps, and adoption of specially designed gradient elution and a special eluent, YJM A and YJM B with the purity exceeding 97% can be obtained, and the establishment of the preparation process greatly reduces the separation and purification cost of Yuanjiang mycin. The anti-tumor performance of the YJM A disclosed by the invention is far better than that of similar compounds.

Description

technical field [0001] The invention relates to a yuanjiangmycin analogue, a preparation method thereof and an antitumor application, and belongs to the technical field of biomedicine. Background technique [0002] Actinomycetes are an important source of novel bioactive molecules with diverse pharmacological properties, including antibiotic, antifungal, antiparasitic, and anticancer drugs. Although most of the skeletons in use today were discovered between 1940 and 1960, with sequencing technology and bioinformatics analysis, the biosynthetic potential of actinomycetes may be underestimated because almost each actinomycete has more than 20 biological Synthetic gene clusters. However, under laboratory conditions, most clusters are often not expressed or expressed very little, which hinders the discovery of new compounds. Therefore, using new methods and technologies to activate microbial silencing gene clusters and exploring new natural products is one of the important dir...

Claims

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Application Information

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IPC IPC(8): C07D493/18A61P35/00C12P17/18C12R1/465
CPCC07D493/18C12P17/181A61P35/00Y02A50/30
Inventor 段燕文朱湘成庄周康
Owner HAYAO CIHANG PHARM CO LTD
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