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Use of green chiretta diterpene lactone in inhibiting vascularization

A technology of andrographis diterpene lactone and diterpene lactone is applied in the new application field of andrographis diterpene lactone, which can solve the problem that the anti-cancer mechanism of active ingredients is unclear, the anti-cancer effect is limited, the dosage is unreasonable, and the like. problems, to achieve the effect of not easy drug resistance, high curative effect, and stable expression

Inactive Publication Date: 2008-11-05
PEKING UNIV FIRST HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although Andrographis paniculata has been used in clinical treatment of tumors and has a certain curative effect, the anti-cancer mechanism of its active ingredients is not clear and the dosage is unreasonable, which limits its anti-cancer effect.

Method used

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  • Use of green chiretta diterpene lactone in inhibiting vascularization
  • Use of green chiretta diterpene lactone in inhibiting vascularization
  • Use of green chiretta diterpene lactone in inhibiting vascularization

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: The inhibitory effect of andrographis paniculata diterpene lactone on the proliferation of human vascular endothelial cells.

[0032] Human umbilical vein endothelial cells (HUVEC, purchased from Cascade Biologics, Cot: C-003-5C) were cultured in Medium 200 (purchased from Cascade Biologics) containing 10% fetal bovine serum (FBS) (37°C, 5%CO 2 , 95% humidity) culture, the fourth-generation cells were inoculated in a 96-well culture plate at a density of 4000 / 250ul, and a control group, a drug group and a blank control group with various concentration gradients were set, and 3 replicate holes were made in each group. When the cell growth density reaches 80%, add 5ul of four kinds of andrographolide diterpene lactone compounds (i.e.: andrographolide, dehydroandrographolide, deoxyandrographolide, neoandrographolide) of each concentration gradient respectively, all dissolved with DMSO ), so that the final concentration was 1000uM, 100uM, 10uM, 5uM and 1uM respe...

Embodiment 2

[0033] Example 2: Inhibitory effect of andrographis paniculata diterpene lactone on tube formation ability of human vascular endothelial cells.

[0034] Add BD Matrigel to each well of a 24-well culture plate TM Matrix original glue 200ul, make it polymerized into gel, the fourth generation human umbilical vein endothelial cell (HUVEC) suspension was inoculated into the 24-well plate coated with Matrigel glue at a density of 30000cell / 500ul, set the control group, each concentration gradient The drug group was added with 10ul concentration gradient of andrographolide (200uM, 1000uM, dissolved in DMSO), and the control group was added with 10ul of sterile DMSO, and 3 replicate wells were made in each group. 37°C, 5% CO 2 , 95% humidity for 24 hours, OLYMPUS CK40-RFL optical microscope to observe the formation of vascular endothelial cell tubes, take 4 low-power fields from each well and count them, and take pictures with an OLYMPUS CK40 digital camera. The result is as ima...

Embodiment 3

[0035] Example 3: Inhibitory effect of andrographolide diterpene lactone on migration ability of human vascular endothelial cells.

[0036] 1. Preparation of chemokines

[0037] The NIH3T3 cells that grew well the next day after passage were used. Gently rinse with serum-free DMEM twice. Serum-free DMEM, 5% CO 2 , 37°C for 24-48 hours. Collect the cell supernatant. Centrifuge (12000g, 4°C, 10min). Filter the supernatant (0.22um filter membrane) and store in aliquots (-20°C).

[0038] 2. Matrigel invasion test

[0039] Take 25ul of diluted Matrigel (the original glue was diluted with DMEM at a ratio of 1:2) and add it to the chamber on the Transwell plate to cover the entire surface of the polyester film, and incubate at 37°C for 30min to make Matrigel polymerize into a gel. The fourth-generation human umbilical vein endothelial cell (HUVEC) suspension was inoculated into the upper chamber at a density of 30000 / 250ul, washed 3 times with PBS, digested and harvested from ...

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Abstract

The present invention discloses the new use of andrographolide in inhibiting vasculariztion. Research probes the vasculariztion inhibiting effect of andrographolide. The technological scheme is that andrographolide, dewatered andrographolide, deoxyandrographolide and neoandrographolide are used in preparing medicine for prevention and / or treatment of inhibiting vasculariztion with the dosage being 3.0-50.0, 5.0-50.0, 3.5-45 and 5.0-60 mg / kg body weight / day successively. The present invention uses the vasculariztion as the target of treating diseases and has the advantages of treatment specificity of pointing newly formed vessel; small dosage, high treating effect and less side effect owing to the direct action of medicine to vascular endothelial cell; and less medicine resistance owing to relatively stable endothelial cell gene expression.

Description

technical field [0001] The present invention relates to a new application of the compound, in particular to a new application of andrographis paniculata diterpene lactone. Background technique [0002] Studies have shown that tumors themselves have the ability to initiate and promote angiogenesis to meet their own metabolism and nutrient supply needs. Tumor angiogenesis starts from the existing vascular bed. If there is no angiogenesis, the growth of the primary tumor will not exceed 1-2mm, and there will be no invasion and metastasis. Inhibiting tumor angiogenesis is closely related to blocking tumor occurrence, development, invasion and metastasis. Inhibiting tumor angiogenesis can also prevent the malignant transformation of precancerous lesions into cancer. Not only the growth, invasion and metastasis of solid tumors depend on angiogenesis, but also the growth and metastasis of hematological malignancies (such as malignant lymphoma, lymphocytic leukemia, etc.) are clo...

Claims

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Application Information

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IPC IPC(8): A61K31/365A61P35/00
Inventor 王振军赵博
Owner PEKING UNIV FIRST HOSPITAL
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