Method for preparing PD-1/PD-L1 inhibitor from clematis tangutica

A PD1-PDL1, clematis technology, applied in pharmaceutical formulations, medical preparations containing active ingredients, plant/algae/fungus/moss ingredients, etc., can solve the problems of poor oral administration, high cost, side effects, etc.

Active Publication Date: 2020-07-07
QINGHAI UNIV FOR NATITIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the inherent disadvantages of antibody therapy, such as high cost, poor oral administration, im...

Method used

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  • Method for preparing PD-1/PD-L1 inhibitor from clematis tangutica
  • Method for preparing PD-1/PD-L1 inhibitor from clematis tangutica
  • Method for preparing PD-1/PD-L1 inhibitor from clematis tangutica

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Preparation of Crude Extract of Active Components of Clematis tanguticae

[0046] The preparation method of the active ingredient of Clematis tangutica effectively blocking PD1-PDL1 binding is as follows:

[0047] (1) Extraction: Select high-quality Clematis Tanguta, wash and dry it with pure water, then crush it, first soak it in 60% ethanol for 8 hours, extract twice and filter to obtain the filtrate, recover the solvent under reduced pressure, add 70% ethanol to the filter residue Ultrasonic extraction was performed twice, each time for 3 hours, concentrated under reduced pressure to obtain 70% ethanol extract; extracted once with 83% ethanol in the same way, concentrated under reduced pressure to obtain 83% ethanol extract, and combined to obtain total ethanol extract. The total ethanol extract is completely dissolved in water at 1:3, and then extracted with ethyl acetate, the ratio of water to ethyl acetate is 0.4:1, and the ethyl acetate part and the aqu...

Embodiment 2

[0053] Example 2 Preparation of Crude Extract of Active Components of Clematis tanguticae

[0054] The preparation method of the active ingredient of Clematis tangutica effectively blocking PD1-PDL1 binding is as follows:

[0055] (1) Extraction: select high-quality Clematis Tanguta, wash and dry it with pure water, then crush it, first soak it in 74% ethanol for 4 hours, extract twice and filter to obtain the filtrate, recover the solvent under reduced pressure, add 80% ethanol to the filter residue Ultrasonic extraction twice, 4 hours each time, concentrated under reduced pressure to obtain 80% ethanol extract; extracted once with 85% ethanol in the same way, concentrated under reduced pressure to obtain 85% ethanol extract, combined to obtain total ethanol extract. The total ethanol extract is completely dissolved in water at 1:4, and then extracted with ethyl acetate, the ratio of water to ethyl acetate is 0.5:1, and the ethyl acetate part and the aqueous phase extract par...

Embodiment 3

[0061] Example 3 Performance detection of Clematis tangutata blocking PD1-PDL1 binding active site

[0062] The active part of Clematis tanguticae obtained by extraction and purification in Examples 1 and 2 is used as a drug performance detection substance, and the following experiments are carried out:

[0063] Among them, the HTRF PD1 / PDL1 detection kit (purchased from CISbIO) was used in the experiment. The detection principle of the HTRF PD1 / PDL1 detection kit: the HTRF PD1 / PDL1 detection kit is to detect the protein interaction between PD1 and PDL1. Using HTRF (Homogeneous Time-Resolved Fluorescence) technology, individual compounds and antibody blockers can be characterized quickly, easily and with high throughput. The interaction between Tag1-PD-L1 and Tag2-PD1 was detected by using anti-Tag1-europium (HTRF donor) and anti-Tag2-XL665 (HTRF acceptor). When the donor antibody and the acceptor antibody approach due to the binding of PD-L1 and PD1, the excitation of the do...

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Abstract

The present invention discloses an active ingredient extracted from clematis tangutica as a natural plant and an extraction method of the active ingredient. More particularly the active ingredient extracted from the clematis tangutica can effectively block PD1-PDL1 combination to remove, inhibit, reduce or block PD1-PDL1 interaction and subsequent action; a pharmaceutical composition of the activeingredient and a pharmaceutical carrier can effectively become a novel PD1-PDL1 inhibitor; and the PD1-PDL1 inhibitor is applied to anti-tumor related diseases, so that references are provided for clinical treatment schemes.

Description

Technical field: [0001] The present invention relates to the application technical field of natural products, and extracts active ingredients from natural plant Clematis tangutica, more specifically relates to extracting active ingredients from clematis tangutica that can eliminate, inhibit, reduce or block the combination of PD1-PDL1 As well as the pharmaceutical composition of the active ingredient and the pharmaceutically acceptable carrier, as well as the application on related diseases. Background technique: [0002] The body's immune system can identify tumor cells, regulate tumor growth, and even eliminate tumors, and T cells play a central role in this process. The activation of T cells first requires the first signal stimulation provided by antigen-presenting cells, and also requires the second signal stimulation provided by co-stimulatory molecules. Costimulatory molecules not only provide co-stimulatory signals to enhance immunity, but also provide co-inhibitory ...

Claims

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Application Information

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IPC IPC(8): A61K36/716A61P35/00A61P35/02
CPCA61K36/716A61P35/00A61P35/02A61K2236/333A61K2236/39A61K2236/51A61K2236/55
Inventor 尕藏扎西林鹏程吴疆潘国庆张金魁王琪叶萍
Owner QINGHAI UNIV FOR NATITIES
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