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PD-L1 inhibitor and preparation method thereof

A PD-L1 and preparation technology, applied in the field of biopharmaceuticals, can solve problems such as poor curative effect, achieve the effect of improving tumor immunity, good inhibitory effect and therapeutic effect, and reducing interaction

Inactive Publication Date: 2019-03-12
RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In view of the above-mentioned defects of the prior art, the purpose of the present invention is to address the deficiencies of the prior art and the demand for treating tumors, and solve the existing immune response based on the combination of PD-L1 antibody and PD-L1 to activate T cells, so as to achieve There are problems such as poor curative effect in the method of improving anti-tumor immune activity

Method used

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  • PD-L1 inhibitor and preparation method thereof
  • PD-L1 inhibitor and preparation method thereof
  • PD-L1 inhibitor and preparation method thereof

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preparation example Construction

[0038] The invention provides a preparation method for inhibiting the expression of PD-L1 protein and reducing the localization and distribution of PD-L1 protein in the cell membrane, comprising the following steps:

[0039] Step 1, dissolving 2-bromopalmitic acid in a solvent, preparing a concentrated solution of 2-bromopalmitic acid, subpackaging and storing in a freezer;

[0040] Step 2, adding fetal bovine serum with a weight percentage of 10% to 20% into the medium to prepare a medium containing fetal bovine serum;

[0041] Step 3, adding the 2-bromopalmitic acid concentrate obtained in step 1 to the medium containing 10% to 20% by weight of fetal bovine serum in step 2, to obtain the inhibition of PD-L1 protein expression and the reduction of PD- A preparation for the localization and distribution of L1 protein in the cell membrane.

[0042] In a preferred embodiment of the present invention, in step 1, the solvent is a water-miscible solvent, preferably ethanol and dim...

Embodiment 1

[0056] Example 1. Preparation of a preparation (2-BP) that inhibits the expression of PD-L1 protein and reduces the localization and distribution of PD-L1 protein in the cell membrane

[0057] Step 1. Dissolve 2-bromopalmitic acid in ethanol at a molar concentration of 10 mM to prepare a concentrated solution of 2-bromopalmitic acid, keep it away from light, store it at -60°C to -80°C after subpackaging, and avoid repeated freezing and thawing;

[0058] Step 2, adding 10% to 20% fetal bovine serum by weight into the DMEM medium to prepare a medium containing 10% to 20% fetal bovine serum;

[0059] Step 3, the 2-bromopalmitic acid concentrated solution that step 1 obtains is added in the culture medium that contains the fetal calf serum of weight percentage composition 10%~20% in step 2, obtains 2-bromopalmitic acid molar concentration (mol / liter ) is a 10uM preparation (2-BP) that inhibits the expression of PD-L1 protein and reduces the localization and distribution of PD-L1 p...

Embodiment 2

[0060] The preparation of embodiment 2, 2-BP

[0061] Step 1. Dissolve 2-bromopalmitic acid in dimethyl sulfoxide at a molar concentration of 80mM to prepare a concentrated solution of 2-bromopalmitic acid, keep it away from light, store it in a freezer at -80°C to -100°C after subpackaging, and avoid repeated freezing and thawing;

[0062] Step 2, adding 5% to 10% fetal bovine serum by weight into the RPMI1640 medium to prepare a medium containing 5% to 10% fetal bovine serum;

[0063] Step 3, step 1 obtains 2-bromopalmitic acid concentrated solution and joins in the culture medium that contains the fetal bovine serum of weight percentage composition 5%~10% in step 2, obtains 2-bromopalmitic acid molar concentration (mol / liter ) is an 80uM preparation (2-BP) that inhibits the expression of PD-L1 protein and reduces the localization and distribution of PD-L1 protein in the cell membrane.

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Abstract

The invention discloses a preparation for inhibiting PD-L1 protein expression quantity in tumor cells and reducing positioning distribution of PD-L1 proteins in cell membranes. The preparation comprises 2-bromine palmitate with the molar concentration of 10-100 micromoles, fetal calf serum and a culture medium. A preparation method includes the steps: preparing 2-bromine palmitate concentrated solution and the culture medium containing the fetal calf serum; adding the 2-bromine palmitate concentrated solution into the culture medium containing the fetal calf serum. According to the preparationfor inhibiting PD-L1 protein expression quantity in the tumor cells and reducing positioning distribution of the PD-L1 proteins in the cell membranes, expression of the PD-L1 proteins in the tumor cells can be reduced, positioning of the PD-L1 in tumor cell membranes is reduced, the preparation has double PD-L1 inhibiting effects, immune response of T cells can be activated, tumor immunity is improved, and the preparation is widely applied to PD-L1 regulation biomedicine and pharmaceutical research.

Description

technical field [0001] The invention relates to the technical field of biopharmaceuticals, in particular to a preparation for inhibiting PD-L1 protein expression and cell membrane localization and a preparation method thereof. Background technique [0002] Malignant tumors are a type of disease with the highest morbidity and mortality in my country. In recent years, the development of clinical diagnosis, surgery, radiotherapy and chemotherapy has enabled some patients with malignant tumors to receive early detection and effective treatment. However, searching for new treatment methods and drugs has long been a hot spot in the field of cancer research all over the world. Different from traditional treatment methods, tumor immunotherapy can activate or induce the body to establish a specific immune response to tumor antigens, eliminate primary or metastatic tumor cells, establish immune memory, and prevent tumor recurrence, drug resistance and metastasis. Programmed cell deat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/20A61P35/00A61P37/04
CPCA61K31/20A61K35/16A61K2300/00
Inventor 许杰姚晗王焕彬李楚舒章瑶石虎兵房静远陈萦晅
Owner RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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