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Medicinal composition and application thereof

A composition and drug technology, applied in the direction of drug combination, anti-tumor drug, pharmaceutical formulation, etc., can solve the problems of no literature discloses liver cancer, no literature reports the synergistic effect of human endostatin and dexamethasone, etc., and achieves high antitumor effect. Liver cancer activity, the effect of high-efficiency preselected drugs

Active Publication Date: 2012-07-25
SHANDONG SIMCERE BIO PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, there is no literature that discloses that the combined use of human endostatin and dexamethasone has an effect on liver cancer, and there is no literature that gives technical inspiration for such combined use, and there is no literature that reports the combined use of human endostatin and dexamethasone synergistic

Method used

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  • Medicinal composition and application thereof
  • Medicinal composition and application thereof
  • Medicinal composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1 Clone formation analysis The influence of composition of the present invention on the proliferation of human umbilical vein endothelial cells (HUVEC)

[0041] Human umbilical vein endothelial cells (HUVECs) were diluted with ECM medium and inoculated in 24 wells (100cells / well), incubated at 37°C for 3 days and then treated with drugs. The experiment was divided into control group (Control, that is, serum-free ECM medium, The same below), recombinant human endostatin group (abbreviated as Endo group below and in the figure, recombinant human endostatin (abbreviated as Endo below): 12.5 μg / ml), dexamethasone sodium phosphate group (abbreviated as Endo below and in figure It is abbreviated as DXM group, dexamethasone sodium phosphate (hereinafter abbreviated as DXM): 0.5 μg / ml) and combination group (Endo12.5 μg / ml+DXM0.5 μg / ml). Continue to cultivate for 10 days, observe under the microscope and calculate the number of colony formation ( figure 1 ). The cl...

Embodiment 2

[0049] Example 2 Transwell method to analyze the effect of the composition of the present invention on the invasion of HUVECs

[0050] Place the Transwell chamber in a 24-well plate, equilibrate each chamber overnight with serum-free medium, then coat the upper layer of the chamber with Matrigel gel, inoculate HUVEC cells in the upper chamber of the Transwell, add ECM medium in the lower chamber, and then Adding drugs and treating at 37°C for 18 hours, the experiment was divided into control group (Control), Endo group (250 μg / ml), DXM group (5 μg / ml) and combined group (Endo250 μg / ml+DXM5 μg / ml). Take out the small chamber and stain with hematoxylin, erase the cells that have not passed through the membrane, take pictures and count the cells that have penetrated the membrane ( figure 2 ), and calculate the cell invasion rate and synergy index (CDI) according to the following formula:

[0051] Cell invasion rate = number of penetrating cells in the treatment group / number of ...

Embodiment 3

[0055] Example 3 Tube Formation Experiment Analysis of the composition of the present invention on the impact of HUVEC tubule formation

[0056] HUVEC cells were seeded in 100 μl Matrigel-coated 96-well plates (2×10 4 cells / well), and dosing treatment. The experiment was divided into control group (Control), Endo group (250μg / ml), DXM group (5μg / ml) and combination group (Endo250μg / ml+DXM5μg / ml). After incubation at 37°C for 24 hours, photographs were taken under an inverted microscope (×200), and the tubular structures formed by HUVEC cells were counted ( image 3 ), and calculate the inhibition rate and synergy index according to the following formula:

[0057] Inhibition rate=[1-(the number of tubules formed in the treatment group / the number of tubules formed in the control group)]×100%;

[0058] CDI=AB / (A×B), A=the number of tube formation in Endo group / the number of tube formation in control group, B=the number of tube formation in DXM group / the number of tube formatio...

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Abstract

The invention belongs to the technical field of biological products and discloses a medicinal composition and application thereof. The composition takes recombinant human endostatin and hexadecadrol as effective constituents, wherein the mole ratio of the recombinant human endostatin to the hexadecadrol is 10-100:1. The recombinant human endostatin in the combination can co-act with the hexadecadrol to restrain the multiplication, invasion, migration and angiogenesis of endothelial cells. An in-vivo test result proves that the recombinant human endostatin and the hexadecadrol co-act with eachother to restrain the growth of the mice H22 transplanted liver tumor and human Bel7402 transplanted liver tumor. Therefore, the composition can be widely applied to medicaments for treating liver cancer.

Description

technical field [0001] The invention belongs to the technical field of biological products, and relates to a pharmaceutical composition and its application. Background technique [0002] Human vascular endostatin (endostatin) is an enzyme-cleaved product with a molecular weight of 20kDa at the carboxyl end of collagen x VlII, which has the activity of inhibiting the proliferation and migration of vascular endothelial cells, and inhibiting angiogenesis in vivo. It is the strongest endogenous protein known so far. Sexual angiogenesis inhibitory factor. [0003] Recombinant human endostatin is a protein with angiogenesis inhibitory activity obtained by cloning, expressing and purifying the human endostatin gene with prokaryotic or eukaryotic expression vectors using DNA recombination engineering technology. [0004] The Chinese invention patent "Method for Producing Endostatin" (patent publication No. CN132481 8A, publication date is December 5, 2001) provides an improved prod...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/17A61P35/00A61K31/573
Inventor 甄永苏李兴起尚伯扬王冬春张胜华吴淑英
Owner SHANDONG SIMCERE BIO PHARMA CO LTD
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