Application of MCT1 inhibitor and pharmaceutical composition containing the MCT1 inhibitor

A technology of MCT1 and inhibitors, which is applied in the field of application and pharmaceutical compositions containing it, can solve the problems such as incompletely clear regulatory mechanism, and achieve the effects of promoting growth, reducing expression and promoting proliferation

Inactive Publication Date: 2020-08-07
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In view of the key role played by the abnormal glucose metabolism of cancer cells in the proliferation of cancer cells, the regulatory mechanism of MCT1 in ovarian cancer is not completely clear. In-depth research on this mechanism is a very important scientific issue.

Method used

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  • Application of MCT1 inhibitor and pharmaceutical composition containing the MCT1 inhibitor
  • Application of MCT1 inhibitor and pharmaceutical composition containing the MCT1 inhibitor
  • Application of MCT1 inhibitor and pharmaceutical composition containing the MCT1 inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] In this example, SKOV3 ovarian cancer cells were transfected by constructing a pcDNA3.1-MCT1 eukaryotic expression vector, and Western Blot was used to detect the expression level of MCT1 and the content of lactic acid in the culture medium. Specifically:

[0035] 1. Preparation of separation gel (lower layer glue): the glass plate is cleaned by the following steps: cleaning with washing liquid, rinsing with tap water, ddH 2 O Rinse, dry in the air, and bake in a 68°C oven. Take out the glass plate, and install the glass glue plate after cooling to room temperature. The concentration of acrylamide in the lower gel depends on the molecular weight of the protein to be separated, generally 8-10%, which can separate most of the proteins. Prepare a separating gel according to the SDS-PAGE gel formulation table, then fill the gel with a 1000μL sample gun, and then use n-butanol or ddH 2 O pressure glue to ensure the smooth surface of the glue. At room temperature, after a...

Embodiment 2

[0043] MTT method to detect cell proliferation

[0044] 1. Cell inoculation: Take the cells in the logarithmic growth phase and make a single cell suspension with the culture medium containing 10% fetal calf serum, and use 1×10 per well 4 Cells were seeded into a 96-well plate with a volume of 200 μl per well at 37°C in 5% CO 2 Cultivate in the incubator for 24h.

[0045] 2. Treat cells with different concentrations of MCT1 inhibitor AR-C155858 (0, 10, 50, 100, 200 μmol / L), 37 ° C, 5% CO 2 Continue culturing for 24 h in the incubator.

[0046] 3. Coloring: Add 20 μl of MTT solution (5 mg / ml in PBS) to each well of the cultured cells, continue to incubate for 4 hours, stop the culture, carefully suck out the culture supernatant in the well, and then centrifuge for the suspension cells Aspirate and discard the culture supernatant in the well. Add 150 μl DMSO to each well and shake for 10 minutes to fully melt the crystals.

[0047] 4. Colorimetry: select a wavelength of 490...

Embodiment 3

[0051] lipofection

[0052] According to the liposome transfection procedure of Invitrogen Company, miRNA was dissolved in a certain volume of serum-free Opti-MEM medium, and mixed evenly. Add an appropriate amount of Lipofectamine 2000 to an equal volume of serum-free Opti-MEM, and mix well. Incubate at room temperature for 5 minutes and then drop the former mixture into the latter mixture dropwise, shaking while dropping, so that the above two mixtures are mixed evenly. Incubate for 25 minutes at room temperature. Aspirate the culture medium from the dish, then add the transfection mixture. Cells at 37°C, 5% CO 2 After culturing in the incubator for 6 hours, replace it with normal complete medium.

[0053] Routinely cultured ovarian cancer cells, chemically synthesized (Guangzhou Ruibo Bio) can bind to MCT1 miRNA-199a-5p and miRNA-199a-5p-inhibitor, miRNA-29c and miRNA-29c-inhibitor, miRNA-27a-3p and miRNA- 27a-3p inhibitor, miRNA-506-3p and miRNA-506-3p-inhibitor were ...

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Abstract

The invention discloses application of an MCT1 inhibitor and a pharmaceutical composition containing the MCT1 inhibitor, and belongs to the technical field of biological medicines. According to the application of the MCT1 inhibitor in preparation of the anti-ovarian cancer medicine, the effect of resisting ovarian cancer is achieved by reducing the expression level of MCT1 in ovarian cancer cells;a microRNA regulation and control mechanism of MCT1 is discovered for the first time, wherein the MCT1 can effectively promote proliferation of tumor cells; so that through the MCT1, the tumor cellscan exclude lactic acid out of the cells, and growth of the tumor cells is facilitated. The miRNA199a-5p provided by the invention can effectively participate in the regulation and control of MCT1 andinhibit the expression of MCT1.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to the application of an MCT1 inhibitor and a pharmaceutical composition containing it. Background technique [0002] Dysregulation of energy metabolism in malignant tumors is one of its important malignant features, involving changes in the expression and activity of various genes. Cancer cells carry out the metabolic process of glycolysis even under aerobic conditions, which is called "Warburg effect". The high energy consumption state of rapid proliferation of cancer cells and the low production efficiency of glycolysis form a pair of contradictions. This requires cancer cells to take in and consume a large amount of glucose to provide the energy they need, and the incomplete degradation products of glycolysis also provide NADPH as a carbon source for the proliferation of cancer cells. The final product of glycolysis, pyruvate, does not enter the tricarboxylic a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K31/7088A61P15/08A61P35/00
CPCA61K45/00A61K31/7088A61P15/08A61P35/00
Inventor 杨红李郁李佳宋婷婷徐盈程璐陈柳
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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