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Preparation method of venenum bufonis extract and application thereof in preparation of anti-brain glioma medicine

A technology of extract and toad venom, applied in the field of traditional Chinese medicine extraction, can solve the problems of unsatisfactory curative effect and poor prognosis

Inactive Publication Date: 2016-07-06
赵婷
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, chemical drugs and general anti-tumor traditional Chinese medicines are difficult to kill glioma cells and penetrate the blood-brain barrier at the same time, and the curative effect is not satisfactory. As a result, glioma is still the worst prognosis among systemic tumors. one of the tumors

Method used

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  • Preparation method of venenum bufonis extract and application thereof in preparation of anti-brain glioma medicine
  • Preparation method of venenum bufonis extract and application thereof in preparation of anti-brain glioma medicine
  • Preparation method of venenum bufonis extract and application thereof in preparation of anti-brain glioma medicine

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0036] The preparation method of toad venom extract of the present invention includes the following steps:

[0037] (1) The toad venom medicinal material is crushed into coarse powder, which is over 60 mesh sieve, and dissolved in ethanol with a volume concentration of 80%. The mass of ethanol is 10 times the mass of coarse powder. The dissolution is heated in a water bath and the heating temperature is 80%. ℃, heating time 1h; then suction filtration, evaporate the ethanol of the filtrate, and dry to obtain a crude extract.

[0038] (2) Add the crude extract to distilled water, where the mass of the distilled water is 10 times the mass of the crude extract, and ultrasonically suspend to obtain a suspension; then add ethyl acetate for extraction, where the mass of ethyl acetate is 10 times the mass of the suspension. After doubling, the extract is obtained; the ethyl acetate in the extract is evaporated and dried in vacuum to obtain the extract.

[0039] (3) Dissolve the extract in ...

experiment example 2

[0045] The above-mentioned common solution (bufadienolides solution, BU-S) of toad venom extract is used for anti-glioma drugs. The standard dose of 5 mL is: 2.5 mg of toad venom extract, 2.5 mL of propylene glycol as the first solvent, and the rest as second Solvent injection water. Wherein, the combined content of bufalin, bufastin and lipobufagin in the toad venom extract is greater than 95%. This experimental example 2 uses the toad venom extract prepared in experimental example 1.

[0046] The preparation method of the above-mentioned common solution of toad venom extract includes the following steps: weigh 2.5 mg of ding toad venom extract into a 5 mL measuring flask, add 2.5 mL of propylene glycol, dissolve by ultrasound, dilute to 5 mL with water for injection, shake well, and use 0.22 It is obtained by filtering with a μm filter membrane.

experiment example 3

[0048] It shows the inhibitory effect of the above-mentioned common solution (BU-S) of Bufo venom extract on the proliferation of human glioma cell U87-MG in vitro.

[0049] (1) Cell culture: U87-MG glioma cells are placed in DMEM medium containing 10% fetal bovine serum by volume and placed in CO at 37°C and 5% by volume 2 Culture in an incubator, when the cell mass grows to 80%-90% (logarithmic growth phase), digest with 0.25% by mass trypsin and 0.02% by mass ethylenediamineetraacetic acid (EDTA), wash, and resuspend Suspend and dilute to 1x10 5 Cells / ml single cell suspension, inoculated into 96-well plates, placed in an incubator for 24 hours, discarded the culture medium, and added BU-S complete cell culture medium containing different concentrations (1.625μg / ml, 3.25μg / ml, 6.5μg / ml, 15μg / ml, 30μg / ml, 60μg / ml) and incubated for different times (24h, 48h, 72h), the cell viability was determined by the tetramethylazazole blue method (MTT) .

[0050] The result is figure 1 As ...

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Abstract

The invention relates to the technology of traditional Chinese medicine extraction and discloses a preparation method of a venenum bufonis extract, by which the venenum bufonis extract being higher than 95% in comprehensive content of bufalin, cinobufagin and resibufogenin can be prepared. The invention also discloses an application of the venenum bufonis extract being higher than 95% in the comprehensive content of the bufalin, cinobufagin and resibufogenin in preparation of an anti-brain glioma medicine.

Description

Technical field [0001] The invention relates to Chinese medicine extraction technology, in particular to a preparation method of toad venom extract and its application in preparing anti-glioma medicine. Background technique [0002] Glioma is one of the most common primary brain tumors caused by the canceration of glial cells in the brain and spinal cord. It accounts for about 35% to 61% of intracranial tumors. It is highly aggressive and has a poor prognosis. It is death Malignant tumors with a high rate and disability rate. At the beginning of the onset of glioma, there are usually no typical symptoms, and the invasive growth characteristics of glioma located in important functional areas and deep parts make it difficult to complete the resection, not very sensitive to radiotherapy and chemotherapy, and very easy to relapse. However, chemical drugs and general anti-tumor Chinese medicines have difficulty in killing glioma cells and penetrating the blood-brain barrier at the sa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/65A61P35/00
Inventor 赵婷
Owner 赵婷
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