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Preparation method of efficient antitumor antibiotic calicheamicin

A calicheamicin, polystyrene technology, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problem of insufficient expression of biosynthetic genes, low calicheamicin yield, Significant inhibition of bacterial metabolism and other problems, to achieve the effect of eliminating toxic effects, reducing energy consumption, and stabilizing the process

Inactive Publication Date: 2012-10-17
ZHEJIANG HISUN PHARMA CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the fermentation production is carried out according to the culture conditions described in the literature, which is affected by the characteristics of the producing bacteria, the fermentation process, and the control conditions, especially the high toxicity of the product, and the synthesis of the product significantly inhibits the metabolism of the bacteria, so that its biosynthesis genes cannot be obtained. Fully expressed, resulting in low yield and high cost of calicheamicin, making it difficult to form industrialization

Method used

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  • Preparation method of efficient antitumor antibiotic calicheamicin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1 prepares calicheamicin

[0034] (1) Preparation and cultivation of slant spores

[0035] Slant spore medium formula (g / L): Yeast extract 4, malt extract 10, glucose 4, agar 20, pH 7.2 before elimination, test tube 30×200mm, capacity 20mL, sterilized at 121°C for 20 Minutes later, cool down to 50-60°C and place an inclined plane, insert the spores and cultivate them for 12-14 days at 28±1°C, the spores mature.

[0036] (2) Preparation and cultivation of seed solution

[0037] Seed medium formula (g / L): soluble starch 24, glucose 5, peptone 5, soybean cake powder 3, CaCO 3 4. The pH before disinfection is 7.0, the volume of the shaker bottle is 250 mL in a Erlenmeyer bottle, and it is sterilized at 121 °C for 20 minutes. The spore inoculum size is 10 5 -10 6 c.f.u. / mL, culture temperature 28±1° C., 250 rpm, shaker culture for 36-48 hours, at this time the pH of the culture solution is 7.0-7.5, and the concentration of mycelium is 10-15% (volume percenta...

Embodiment 2

[0040] Embodiment 2 prepares calicheamicin

[0041] The preparation of slant spores and the preparation of seed liquid are shown in Example 1, the fermentation medium formula is shown in Example 1, wherein the resin is replaced by H-41 resin 30g / L, the fermentation conditions are shown in Example 1, and the calicheamicin output is 40mg / L.

Embodiment 3

[0042] Embodiment 3 prepares calicheamicin

[0043] The preparation of slant spores and the preparation of seed liquid are shown in Example 1, the fermentation medium formula is shown in Example 1, wherein the resin is replaced by HZ-803 resin 30g / L, the fermentation conditions are shown in Example 1, and the calicheamicin output is 10mg / L.

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Abstract

The present invention provides a fermentation method of high-yield antitumor antibiotic calicheamicin. The method of the invention employs Micromonospora echinospora for fermentation production of calicheamicin in a fermentation medium added with different kinds of macroporous adsorption resins. The method of the present invention has stable process and substantially increased fermentation yield.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals, in particular to a method for fermenting high-yield antitumor antibiotic calicheamicin. Background technique [0002] Calicheamicin (calicheamicin) is an enediyne antibiotic, and its main active ingredient is calicheamicin g 1 1 . Calicheamicin has strong cytotoxicity and anti-tumor activity, and can be embedded in the minor groove of the DNA double helix, causing DNA double-strand breaks and killing cells. Calicheamicin has a strong killing effect on a variety of tumor cells, and is the most active class of anti-tumor antibiotics found so far. Due to its strong anti-tumor effect and small molecule size, calicheamicin has been used as a "warhead" of monoclonal antibodies in clinical applications. Among them, the targeted therapy drug Gemtuzumab ozogamicin (alias: GO, Mylotarg, CMA-676), which uses CLM as the "warhead", has been approved by the FDA for the treatment of acute myeloid leukemia....

Claims

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Application Information

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IPC IPC(8): C12P13/02C12R1/645
Inventor 陈华郑玲辉姜璐琰王玲萍白骅
Owner ZHEJIANG HISUN PHARMA CO LTD
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