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Application of protease tablysin to preparation of medicament for treating tumor

A kind of protease and anti-tumor technology, which is applied in the new application field of anti-tumor drugs, can solve the problems that have not been reported in the application, and achieve the effect of inhibiting adhesion and proliferation, good anti-tumor effect, and inhibiting angiogenesis

Active Publication Date: 2012-10-17
JIANGSU CAREFREE PHARM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Studies have found (Dongying Ma, Ren Lai. Anti-thrombosis Repertoire of Blood-feeding Horsefly Salivary Glands. Molecular & Cellular Proteomics, 2009: 2071-2079.), this protease has good thrombolytic and antithrombotic activities, and can be applied to blood embolism The treatment of diseases, there are no application reports in other disease fields

Method used

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  • Application of protease tablysin to preparation of medicament for treating tumor
  • Application of protease tablysin to preparation of medicament for treating tumor
  • Application of protease tablysin to preparation of medicament for treating tumor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1, the influence of Tablysin on the adhesion of human dermal microvascular endothelial cells (MVEC) and matrix protein

[0021] 1.1 Materials and instruments:

[0022] Human dermal microvascular endothelial cells (MVEC) were purchased from Clonetics (San Diego, CA), calf serum FBS (invigrogen, US), trypsin (Cambrex, MD), poly-VTNC (Molecular Innovations, Southfield, MI), fibronectin Protein (Sigma Co, Saint Louis), soluble collagen III (BD Biosciences, Bedford, MA), 96-well fluorescent plate (Thermo Electron Co., Milford, MA, USA), Spectramax Gemini XPS fluorescent plate reader (Molecular Devices , Sunnyvale, CA, USA)

[0023] 1.2 Test method:

[0024] Use EBM-2 culture medium for MVEC cells at 37°C, 5% CO 2 cultured in a cell culture incubator. EBM-2 was supplemented with 2% FBS and single quotes (fibroblast growth factor B, vascular endothelial growth factor, R3 insulin growth factor, ascorbic acid, epidermal growth factor, hydrocortisone and gentamicin...

Embodiment 2

[0031] Embodiment 2, the influence of Tablysin on the proliferation of MVEC

[0032] 2.1 Materials and instruments:

[0033]Human dermal microvascular endothelial cells (MVEC) were purchased from Clonetics (San Diego, CA), calf serum FBS (invigrogen, US), trypsin (Cambrex, MD), and 96-well cell culture plates (Corning Inc., Corning, NY) , CellTiter 96 aqueous non-radioactive cell proliferation assay kit (Promega, Madison, WI), Versamax microplate reader (Molecular Devices, San Diego, CA).

[0034] 2.2 Test method:

[0035] After MVEC was digested with trypsin, 3.2×10 3 The density per well was inoculated in a 96-well cell culture plate, and cultured in EBM-2plus medium (EBM-2 medium containing FBS and single quotes) for 1 day. The next day, the EBM-2plus culture medium was replaced with 200ul of fresh EBM-2 culture medium containing different concentrations of tablysin (0-3μM) and supplemented with 2% FBS and FGF (no other single quotes added). After 3 days, 40ul of MTS so...

Embodiment 3

[0041] Embodiment 3, the impact of Tablysin on the angiogenesis of chicken embryo allantoic membrane in vivo

[0042] 3.1 Materials and instruments:

[0043] Fertilized eggs with white skin were purchased from Qianyuanhao Nanjing Biopharmaceutical Factory, gelatin sponge was purchased from Nanjing Jinling Pharmaceutical Factory of Jinling Pharmaceutical Co., Ltd., and AI RTECH ultra-clean workbench (Antai Company of Suzhou Sujing Group).

[0044] 3.2 Test method:

[0045] 3.2.1 Incubation of breeding eggs: select breeding eggs with clean surface, homogeneous shell, 50-65g in weight, standard shape, uniform air chamber and stomata as the test object. Before incubation, wash with warm water (40-50°C) twice, soak and dry with 1:1000 bromogeramine solution. Place the eggs in an incubator (37°C±5°C) to incubate, with the blunt end facing upwards, and place them on a 6-well culture plate at an angle of 45°C. A wet tray is built into the incubator to keep the relative humidity at 6...

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Abstract

The present invention relates to novel application of a protease tablysin in medicine. The protease tablysin can obviously inhibit adhesion and proliferation effects of human dermal microvascular endothelial cells (MVEC) and the angiogenesis in chicken embryo chorioallantoic membrane. According to research on three common tumor models, the protease tablysin has good antitumor effects, especially for liver cancer, lung cancer and gastric cancer, and can be used in preparation of antineoplastic medicine.

Description

technical field [0001] The invention relates to a new application of protease tablysin in medicine, in particular, it has the effect of anti-tumor angiogenesis and its new application in anti-tumor drugs. Background technique [0002] Tumor angiogenesis has many similarities with angiogenesis under physiological conditions, but there are also great differences, mainly manifested in its uncontrolled and immature nature. The process of tumor angiogenesis is largely regulated by angiogenic factors and angiogenesis inhibitors. Tumor cells, endothelial cells, and macrophages synthesize and release the above-mentioned factors under the influence of hypoxia, systemic stimulation and other factors that change the local microenvironment. When the balance between the two is broken, angiogenesis occurs (Hanahan D , Folkman J. Patterns and emerging mechanisms of the angiogenic switch during tumorigenesis. Cell, 1996, 86: 353-364.), usually includes the following steps: (1) Endothelial ...

Claims

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Application Information

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IPC IPC(8): C12N9/64A61K38/48A61P35/00
Inventor 秦引林
Owner JIANGSU CAREFREE PHARM CO LTD
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