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Application of Incarvillea delavay alkali in preparing drug for resisting autoimmune diseases and graft rejection diseases

An autoimmune disease, pineapple alkaloid technology, applied in metabolic diseases, skin diseases, bone diseases and other directions, can solve the problems of cell non-specificity, increase the economic burden of patients, and high prices, achieve inhibition of occurrence and development, save treatment costs, Source-rich effects

Inactive Publication Date: 2012-11-21
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Existing immunosuppressants mainly contain cyclosporine A (CsA), corticosteroids, rapamycin, etc. Most of them have obvious adverse reactions, such as nephrotoxicity and non-specificity to cells, etc., and the price is high, increasing Patient financial burden

Method used

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  • Application of Incarvillea delavay alkali in preparing drug for resisting autoimmune diseases and graft rejection diseases
  • Application of Incarvillea delavay alkali in preparing drug for resisting autoimmune diseases and graft rejection diseases
  • Application of Incarvillea delavay alkali in preparing drug for resisting autoimmune diseases and graft rejection diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: 30 kg of the whole herb of Incarvillea delavay was extracted 3 times with 200 L of 80% ethanol under reflux, each time for 2 hours, the extracts were combined, concentrated under reduced pressure to obtain 600 g of extract, diluted with water and then diluted with 2% HCl , adjust the pH to 2-3, filter to obtain the filtrate and filter residue, add 20% NaOH to the filtrate to adjust the pH to 11, extract with chloroform to obtain the chloroform fraction; concentrate the extraction fraction under reduced pressure to obtain 250 g of crude product. Mix the crude product with 500g of silica gel, add it to the top of the installed silica gel column, and elute with the gradient of chloroform-acetone (20:1~2:1) system, collect the fraction containing bromelain, and then react with C18 Phase column chromatography, eluting with methanol-water (50%-70%), and thin-layer chromatography detection, obtained 2320 mg of pure bromelain.

Embodiment 2

[0037] Example 2 In vitro culture of human dendritic cells and treatment of DCs with bromelain

[0038] Take 50ml / person of peripheral blood from healthy adults intravenously, anticoagulate with heparin, separate peripheral blood mononuclear cells by density gradient centrifugation, suspend cells in RPMI1640 (1640 medium) containing 10% calf serum, and adjust the cell concentration to 2× 10 6 / ml, add to 24-well culture plate, 0.5ml / well, incubate at 37°C, 5% CO2 for 2 hours, make the mononuclear cells adhere to the wall, wash the culture plate lightly with warm serum-free RPMI1640 to remove the adherence cells, that is, to obtain adherent monocytes. Add serum-containing RPMI1640 containing rhGM-SCF (human recombinant granulocyte colony-stimulating factor) 1000IU / ml and rhIL-4 (human recombinant interleukin 4) 800IU / ml to the culture plate, and incubate at 37°C with 5% CO2 Cultivate in box for 5 days, add bromelain with a final concentration of 2 μmol / L for 24 hours, and the...

Embodiment 3

[0040] Example 3 Effect of bromelain on secretion of IL-10 by human dendritic cells

[0041]According to the method of Example 1, the dendritic cells of the bromelain-stimulated group and the unstimulated group were cultured, the supernatant was collected, and the IL-10 content was detected with an IL-10 ELISA (enzyme-linked immunosorbent assay) kit. The concentration of IL-10 secreted by mononuclear cells transformed dendritic cells in vitro in the pineapple flower stimulation group and the unstimulated group was 435 ± 22pg / ml, 92 ± 14pg / ml, and there was a significant difference (Pfigure 1 ).

[0042] Example 3 Separation of T cells and the effect of bromelain on T cell transformation

[0043] 50ml / person of healthy adult peripheral blood was extracted intravenously, anticoagulated with heparin, and peripheral blood mononuclear cells were isolated by density gradient centrifugation, cultured in RPMI1640 serum-free medium for 2 hours, suspended cells were collected, and after...

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Abstract

The invention provides an application of Incarvillea delavay alkali in preparing a drug for resisting autoimmune diseases and graft rejection diseases. Experiments show that the Incarvillea delavay alkali can inhibit human dendritic cell proliferation in vitro, prevent the human dendritic cells from accelerating the proliferation reaction of allogeneic T cells, and strengthen the capability of the human dendritic cell on secreting interleukin. Meanwhile, in the model of the experimental autoimmune disease cerebrospinal meningitis, the Incarvillea delavay alkali is applied in vivo so as to lower average clinical scores of mice and inhibit the spinal cord infiltration degree of mononuclear cells. In a heart transplantation experiment, the Incarvillea delavay alkali is applied in vivo to prolong the survival time of a transplanted heart. In a delayed hypersensitivity reaction model, the Incarvillea delavay alkali is applied in vivo to lower the auricle swelling degree of the mice. In-vivo and in-vitro results show that the Incarvillea delavay alkali has certain action on resisting autoimmune diseases and graft rejection diseases. The drug provided by the invention is a preparation composed of the Incarvillea delavay alkali which serves as an active ingredient, and a medicinal carrier. The drug can be in an oral agent form, an injection form, a suppository form or an external dosage form and the like.

Description

technical field [0001] The present invention relates to medicines. It specifically relates to the preparation of medicines with new indications, especially the application of bromelain in the preparation of medicines against autoimmune diseases and transplantation rejection diseases. Background technique [0002] Existing immunosuppressants mainly contain cyclosporine A (CsA), corticosteroids, rapamycin, etc. Most of them have obvious adverse reactions, such as nephrotoxicity and non-specificity to cells, etc., and the price is high, increasing patient financial burden. How to use traditional Chinese medicine and its active ingredients to prevent and treat autoimmune diseases and transplant rejection is one of the important directions of modernization research of traditional Chinese medicine. [0003] Bromelain is an alkaloid derived from red pineapple flowers. [0004] At present, it has been found that red pineapple flower can treat postpartum hypogalactia, physical wea...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/435A61P37/00A61P37/06A61P5/16A61P3/10A61P1/16A61P1/00A61P1/04A61P11/00A61P13/12A61P17/00A61P25/00A61P27/02A61P9/00A61P21/04A61P29/00A61P19/02A61P37/08A61P11/06
Inventor 张卫东于益芝张寿德戴先坤曹雪涛苏娟李江燕沈云亨单磊
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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