Application of epigallocatechin gallate combined with tyrosine kinase inhibitor in preparation of cancer treatment medicament

A technology of epigallocatechin and tyrosine kinase, which is applied in the field of biomedicine to achieve the effect of expanding the scope of application

Inactive Publication Date: 2019-07-23
YUNNAN TEAHOME BIOTECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Patients carrying EGFRL858R account for only 6% of all tumor patients, and more than 60% of EGFR wild-type patients have no clinically effective TKI inhibitors available

Method used

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  • Application of epigallocatechin gallate combined with tyrosine kinase inhibitor in preparation of cancer treatment medicament
  • Application of epigallocatechin gallate combined with tyrosine kinase inhibitor in preparation of cancer treatment medicament
  • Application of epigallocatechin gallate combined with tyrosine kinase inhibitor in preparation of cancer treatment medicament

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Effect of EGCG combined with Gefitinib on proliferation of EGFR wild-type and mutant cells in vitro

[0027] 1. Experimental materials

[0028] Cell lines: human epidermal squamous cell carcinoma A431, human lung cancer cell lines NCI-H1666, NCI-H1975.

[0029] 2. Detection principle: MTT method to detect cell viability

[0030] MTT detection cell viability experiment: MTT concentration was 5 mg / ml. Therefore, 0.5 g of MTT can be weighed, dissolved in 100 ml of phosphate buffered solution (PBS) or phenol red-free medium, and filtered with a 0.22 μm filter membrane to remove bacteria in the solution. ℃ and protected from light. Containers are best wrapped in aluminum foil.

[0031] Test steps: A: We use the adherent cell operation method, first collect the logarithmic phase cells to adjust the concentration of the cell suspension, so that the density of the cells to be tested is adjusted to 3 × 10 4 / well, add 200μl to each well, (edge ​​wells are filled w...

Embodiment 2

[0040] Example 2 In the presence of EGCG and EGF at the same time, the effect of Gefitinib on the phosphorylation of EGFR signaling pathway-related proteins in wild-type and mutant cells

[0041] 1. Experimental materials

[0042] Cell lines: human epidermal squamous cell carcinoma A431, human lung cancer cell line NCI-H1975.

[0043] 2. Detection principle: Western Blot detection of changes in related proteins in cells

[0044] 3. Experimental method: a: Treat the cells. After the cells have completed the attachment, starvation treatment is performed, and EGCG and EGF treatment are performed after overnight treatment. Experiments were carried out according to the specific experimental groups. The blank control group was directly added to the acidic medium, and the remaining experimental groups were added to 10mL of the acidic medium to make the final concentration of EGCG 20μg / mL. The EGF mother solution was diluted to 20ng / mL and added to the acidic medium. After completion,...

Embodiment 3

[0050] Example 3 The combined use of EGCG and Gefitinib inhibits the tumor in the EGFR wild-type AA431 cell carcinoma xenograft tumor model

[0051] 1. Experimental materials

[0052] A431 cells and culture: Human epidermal squamous cell carcinoma A431 was cultured in DMEM high-glucose complete medium containing 10% FBS, and the cells were diluted to 2ⅹ106 cells per mL of culture medium. Inoculated into Petri dishes. Placed in a cell culture incubator. Take the cells in the logarithmic growth phase, digest with 0.5% trypsin solution to prepare a single cell suspension, centrifuge at 15000rpm for 3min, discard the supernatant, and adjust the cell density to 5ⅹ10 with PBS 6 / mL.

[0053] Nude mice and their feeding: Male BALB / C nu / nu clean-grade mice, 6-8 weeks, provided by Changzhou Cavens Experimental Animal Co., Ltd., Jiangsu Province. The nude mice were kept in a sterile and ventilated airtight animal room at a temperature of 25°C, with regular light during the day and ...

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Abstract

The invention relates to application of epigallocatechin gallate (EGCG) combined with a tyrosine kinase inhibitor in the preparation of a cancer treatment medicament, and belongs to the technical field of biological medicines. The invention provides a drug administration method combining epigallocatechin gallate (EGCG) as a lead compound with a tyrosine kinase inhibitor, and the combined use can obviously inhibit the growth of EGFR wild-type tumors, and reduce the treatment risk and toxic or side effects caused by anti-cancer drugs in larger doses. The invention improves the sensitivity of theinhibitor and widens the application range of the inhibitor. The invention provides an effective treatment strategy for EGFR wild-type cancer patients.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to an anticancer composition and its application. Background technique [0002] Tyrosine kinase receptor (TKR) and epidermal growth factor receptor (EGFR) are widely expressed on all kinds of cell membranes except suspension cells. The overactivation of EGFR protein is closely related to the occurrence, development, malignancy and prognosis of tumors. EGFR can cause tumor cell proliferation, promote tumor angiogenesis and tumor cell metastasis. EGFR is one of the main targets for targeted therapy of human tumors. Tyrosine kinase inhibitors (TKIs) are a major area of ​​drug development for targeted therapies. Gefitinib, developed for wild-type EGFR tumors, is the first generation of TKI-type EGFR inhibitors, but it was found clinically that Gefiitnib had no significant effect on patients with highly activated EGFR. Studies have found that the affinity of Gefitinb fo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/353A61K31/5377A61P35/00
CPCA61K31/353A61K31/5377A61P35/00A61K45/06A61K38/1808A61P11/00A61K2300/00
Inventor 盛军王宣军黄艳苹字成庭向泽敏黄业伟赵云丽爨向丹徐欢欢罗瑞
Owner YUNNAN TEAHOME BIOTECH CO LTD
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