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Alpha-pinene extracted from pine needle, its extraction method and application for resisting tumor

An extraction method and technology of pinene, applied in α-pinene and its extraction method and its application in anti-tumor, can solve the problems of no obvious symptoms, lack of liver source, poor surgical effect, etc.

Active Publication Date: 2012-02-15
广东源健生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are two main treatment measures that may cure liver cancer: surgical resection or liver transplantation, but due to insidious onset, no obvious symptoms, and advanced liver cancer when discovered, the surgical effect is not good; or due to lack of liver source, surgery is lost Clearance and Liver Transplant Opportunities

Method used

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  • Alpha-pinene extracted from pine needle, its extraction method and application for resisting tumor
  • Alpha-pinene extracted from pine needle, its extraction method and application for resisting tumor
  • Alpha-pinene extracted from pine needle, its extraction method and application for resisting tumor

Examples

Experimental program
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Effect test

Embodiment 1

[0024] Example 1 Extraction and purification of α-pinene

[0025] The tender stems and leaves of local pine needles were naturally dried in the shade, crushed to 40-60 mesh, and 200 g of pine needle powder was taken into a gauze bag by steam distillation, and extracted in a self-designed steam distillation device. Distill for 3 hours at a ratio of 1:8, take the upper essential oil, dissolve it with anhydrous ether, dry it with anhydrous sodium sulfate, and refrigerate for later use. The extracted pine needle oil is transparent yellowish liquid, pine needle oil of aromatic smell. We used the steam fractionation method to separate the pine needle oil from the pine needle extract, and obtained the main components of the pine needle oil as α-pinene and β-pinene, which were detected by high performance liquid phase, and the extraction purity reached 98% ( Figure 1~3 ).

Embodiment 2

[0026] Example 2 Study on the Anti-liver Cancer Effect and Mechanism of α-pinene

[0027] α-pinene can significantly inhibit the proliferation of liver cancer Bel7402 cells:

[0028] Use RPMT-1640 culture medium plus 10% calf serum, inoculate liver cancer 7402 cells in the above culture medium and place in CO 2 37°C in an incubator with 5% CO 2 Culture, digest and collect the cells in the logarithmic growth phase, and adjust the concentration to 5×10 7 / L single cell suspension, seeded in 96-well plate, 0.2ml per well, 24h later, add α-pinene prepared with RPMI-1640 culture medium, the final concentrations were: 8, 2, 0.5, 0.125, 0mg / L (blank control), the experimental groups of liver cancer cells treated with α-pinene at various concentrations, each group was inoculated into 8 replicate wells, placed in the incubator for 24, 48, and 72 h, and 20 μl of MTT solution was added to each well before the culture was terminated. Continue culturing for 4 hours, discard the cultu...

Embodiment 3

[0030] Liver cancer 7402 cells were treated with different concentrations (1:400, 1:600, 1:800, 1:1000) of α-pinene for different times (24h, 48h, 72h), the total RNA was extracted, and SYBR Green real-time fluorescent PCR was used Quantitative techniques were used to examine the expression changes of cell cycle regulatory genes cdc2, CdK1, p53, p21 and cyclinB1. Fluorescent quantitative PCR results showed that α-pinene could significantly down-regulate the expression of cdc25c mRNA in liver cancer Bel-7402 cells ( Figure 5 ).

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Abstract

The invention discloses an alpha-pinene extracted from pine needle, its extraction method and application for resisting tumor. The alpha-pinene of the invention is extracted from pine needle. It is proved by experiments that alpha-pinene has an obvious inhibition effect on the proliferation of hepatoma carcinoma cell. Therefore, the alpha-pinene of the invention can be used in the preparation of drugs for treating tumor related diseases and has great significance.

Description

technical field [0001] The invention relates to the field of extraction and preparation of anticancer drugs, in particular to α-pinene extracted from pine needles, its extraction method and its application in antitumor. Background technique [0002] Liver cancer is a major disease that affects human health. The number of patients in the world exceeds 1 million every year, and its death toll ranks third among tumor deaths. In my country, about 350,000 people are diagnosed with liver cancer each year, and about 320,000 people die from it. At present, there are two main treatment measures that may cure liver cancer: surgical resection or liver transplantation, but due to the insidious onset, no obvious symptoms, and the late stage of liver cancer when discovered, the surgical effect is not good; or due to lack of liver source, surgery is lost Clearance and liver transplant opportunities. Even if there is an opportunity to perform "radical" surgery, the high recurrence and h...

Claims

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Application Information

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IPC IPC(8): C07C13/42C07C7/04A61K31/015A61P35/00
Inventor 陈伟强毛建文李明徐彬魏凤香张丽蓉
Owner 广东源健生物科技股份有限公司
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