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Pseudolaric acid B and application of derivative thereof in preparation of immunity inhibitor

An immunosuppressant, hibiscus acetic acid technology, applied in anti-inflammatory agents, drug combinations, non-central analgesics, etc., can solve the inability to completely prevent the immune response of transplanted tissues, side effects, and inability to completely suppress rejection with tolerated doses, etc. problem, achieve excellent immunosuppressive effect and low toxicity

Inactive Publication Date: 2009-12-23
LOGISTICS UNIV OF CAPF
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Despite its proven efficacy, cyclosporine, as a non-specific immunosuppressant, can lead to reduced immune responses against foreign antigens, and tolerable doses cannot completely suppress rejection
Therefore, the immune response against the transplanted tissue cannot be completely prevented and can cause serious side effects in many organ recipients and exhibit host-dependent effects on the liver, kidneys, central nervous system, and gastrointestinal tract

Method used

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  • Pseudolaric acid B and application of derivative thereof in preparation of immunity inhibitor
  • Pseudolaric acid B and application of derivative thereof in preparation of immunity inhibitor
  • Pseudolaric acid B and application of derivative thereof in preparation of immunity inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Proliferation of T and B Lymphocytes Stimulated by Mitogens

[0035] Aseptically take the mouse spleen, grind it to prepare a single cell suspension, remove red blood cells, and adjust the cell concentration to 5×10 with RPMI-1640 culture medium containing 10% fetal bovine serum (FBS). 6 / mL. Add 100 μl / well of cell suspension in a 96-well plate, and 50 μl / well of different concentrations of samples (hibiscus acetic acid, hexahydro hibiscus acetic acid, deacetylated (R 3 )- hibiscus acetic acid, demethylation (R 2 )- hibiscus acetic acid or deacetylated (R 3 ) Demethylation (R 2 )- Hibiscus acetic acid, BP-1, BP-3, BP-15, BP-20, BP-2, BP-5, BP-6, BP-7, BP-8, BP-10, BP-11, BP-12, BP-13, BP-14, BP-16, BP-17 (BP-18, BP-19) solution, concanavalin A (ConA) or lipopolysaccharide (LPS) of 50 μl / hole, control plus etc. volume medium. After mixing, store at 37°C, 5% CO2 conditions for 48 hours. 4 to 6 hours before the end of the culture, 100 μl of 5 mg / mL MTT s...

Embodiment 2

[0039] Example 2: Unidirectional mixed lymphocyte culture

[0040] Spleen of BALA / c mice was aseptically taken to make a single cell suspension, and the spleen cells of BALA / c mice as stimulatory cells were diluted to 1×10 with complete RPMI-1640 medium. 7 / mL, inactivated with 50 μg / mL mitomycin C in a 37°C water bath, shaken once every 15 minutes, washed three times with PBS, and diluted to 5×10 with complete RPMI-1640 medium 6 / mL. C57BL / 6 mouse splenocytes were used as responding cells. Take 100 μl / well of the above mixture of stimulating cells and responding cells, add different concentrations of hibiscus acetic acid 100 μl / well, add the same amount of medium for the negative control, and add 8×10 -9 M Cyclosporin A (CsA). 37°C, 5% CO 2 Conditioned for 3 days. Added 12h before the end of culture 3 H dilution 25 μl, cells were harvested at the end of incubation, radioactive incorporation was measured by liquid scintillation counting, and data were expressed as pulses...

Embodiment 3

[0051] Example 3: Induction and detection of cytokine production

[0052] Take the spleen of the mouse aseptically, grind it to prepare a single cell suspension, remove the red blood cells, and adjust the cell concentration to 5×10 with RPMI-1640 culture medium containing 10% FBS 6 / mL, cultured in a 96-well plate, 200 μL per well, 3 replicate wells in each group, anti-CD3 monoclonal antibody (anti-CD3) 0.2 μg / mL) was added to the cell suspension, and the drug intervention group was anti-CD3+ different Concentration of Hibiscus acetate arginine salt, negative control plus equal volume medium, positive control plus 8×10 -9 M Cyclosporin A (CsA). 37°C, 5% CO 2 The supernatant was collected after 72 hours of culture under conditions, and the production of IFN-γ was detected by double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). The absorbance was measured at OD450nm with a microplate reader, and the content of cytokines in the samples was converted according to...

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Abstract

The invention discloses a pseudolaric acid B and the application of derivative thereof in the preparation of immunity inhibitor. Experiments prove that the pseudolaric acid B and the derivative thereof have excellent immunity inhibiting effect and lower toxicity, so that the pseudolaric acid B and the derivative thereof or pseudolaric acetate and salt of the derivative of the pseudolaric acid B can be taken as medicine for preventing or treating exclusive reaction, autoimmune disease, inflammatory disease, allergic disease and the like which are related to immunity and caused by organ or skin grafting of mammal (especially human beings).

Description

technical field [0001] The present invention relates to the use of hibiscus acetic acid and its derivatives, in particular to the application of hibiscus acetic acid and its derivatives in the preparation of immunosuppressants. Background technique [0002] Pseudolarix amabilis (Nelson) Regd, a plant in the family Pinaceae, became extinct one after another during the Ice Age of the Pleistocene. Only a few remained in the middle and lower reaches of the Yangtze River in China, and it has become a single genus and single species of endemic plants that only exist in China today. Due to its special taxonomic status, it has become an important research object of plant systems. Although the species of golden pine is rare, its material is fragile, easy to break, and has little economic value. As a result, rare species are not cherished, and forest farmers often destroy them. By selecting some typical forest areas for protective development, the leverage of the market economy can l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/366A61K31/4433A61K31/52A61K31/4178A61K31/427A61K31/496A61P37/06A61P29/00A61P37/08
Inventor 李覃陈虹王伟曹波梅昕白淑芳
Owner LOGISTICS UNIV OF CAPF
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