Limonoids for treating autoimmune diseases

An autoimmune disease, limonin technology, applied in allergic diseases, metabolic diseases, skin diseases, etc., can solve problems such as limited treatment options

Active Publication Date: 2018-08-07
MACAU UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] As the incidence of autoimmune diseases is still rising and treatment options are limited, there is an urgent need for further methods, drugs and compounds to treat autoimmune diseases

Method used

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  • Limonoids for treating autoimmune diseases
  • Limonoids for treating autoimmune diseases
  • Limonoids for treating autoimmune diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0123] Purpose:

[0124] In this experiment, the effects of limonin compound A, limonin compound B, limonin compound C and limonin compound D on the cell survival rate of human peripheral blood mononuclear cells were determined.

[0125] method:

[0126] Human peripheral blood mononuclear cells were isolated from human blood buffy coat provided by Macau Blood Transfusion Center. Specifically, the human blood buffy coat and normal saline were diluted in a 1:1 ratio in a 50 ml centrifuge tube, and then peripheral blood mononuclear cells were obtained by Ficoll density gradient centrifugation. Human peripheral blood mononuclear cells were collected, washed twice with saline and centrifuged at 350g for 10 minutes. These cells can be used for subsequent experiments.

[0127] Cell viability experiment:

[0128] Human peripheral blood mononuclear cells were cultured in RPMI 1640 medium (containing 10% fetal bovine serum (by volume), penicillin (100 units / ml) and streptomycin (100...

Embodiment 2

[0132] Purpose:

[0133] In this experiment, the effects of limonin compound A, limonin compound B, limonin compound C and limonin compound D on ROR-γt luciferase activity were determined.

[0134] method:

[0135] Lightswitch luciferase reporter assay, ROR Gamma T LUCPorterTM Stable Reportercells and LUCPorterTM Vector Control HEK293 cells were cultured in RPMI 1640 medium (containing 10% fetal bovine serum (volume ratio), penicillin (100 units / ml) and streptomycin (100 g / mL) and puromycin (3 g / mL)). They are divided into 5×10 4 cells per well in a 96-well plate. Then 5, 10, 25 and 50 μM of limonin compound A or limonin compound B or limonin compound C or limonin compound D or 10 μM of positive control drug digoxin were added. Incubate in a 37°C incubator for 16 hours, then add the luciferase assay reagent to a 96-well plate, and react in the dark at room temperature for 30 minutes, then transfer the liquid to a white 96-well plate, and detect the fluorescence value with ...

Embodiment 3

[0139] Purpose:

[0140] In this experiment, the effects of limonin compound A, limonin compound B, limonin compound C and limonin compound D on the expression of ROR-γt protein in HEK293 cells were determined.

[0141] method:

[0142] Transfection of ROR-γt plasmid into HEK293 cells

[0143] Transfection experiments were performed according to the instructions of Lipofectamine LTX transfection reagent. HEK293 cells were purchased from American ATCC Company, and the cells were cultured in DMEM medium (containing 10% fetal bovine serum (volume ratio), penicillin (100 units / ml) and streptomycin (100 g / ml)). Take 5×10 5 The cells were seeded in 6-well plates, and 1.5 ml of DMEM medium was added to them for later use. Then 500 microliters of opti-MEM medium was added with 1.25 micrograms of ROR-γt plasmid, plus 1.25 microliters of PLUS reagent, mixed gently, and reacted at room temperature for 5 minutes. Then add LTX reagent, mix gently, and react at room temperature for 30 ...

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PUM

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Abstract

The invention relates to an application of limonoids in preparation of medicine for treating autoimmune diseases, wherein the limonoids include a structure represented as the formula (I), wherein R isoptionally selected from hydroxylated alkyl groups. The invention further relates to an application of the limonoids in preparation of a medicine for inhibiting differentiation of T-helper cell 17, and an application of the limonoids in preparation of a medicine for inhibiting differentiation of T-helper cell 1, T-helper cel 2 or regulative T cells. A method of screening substances capable of inhibiting cells of one or more of Th17, Th1, Th2 and Treg, which can be used for treating the autoimmune diseases. The limonoids can inhibit the differentiation of the Th17, Th1, Th2 and / or Treg cells.

Description

technical field [0001] The present invention relates to a method of treatment of a subject, especially a human, suffering from an autoimmune disease. Autoimmune diseases particularly preferably (but not limited to) include systemic lupus erythematosus, multiple sclerosis, rheumatoid arthritis and the like. The method comprises the therapeutic step of administering to the subject an effective amount of limonin. The present invention further relates to methods of inhibiting the differentiation of T helper cells 17 (Th17), T helper cells 1 (Th1), T helper cells 2 (Th2) and / or regulatory T cells (Treg) and screening methods for inhibiting Th17, Th1 , a method for the differentiation of one or more of Th2 and Treg cells, which can be used for the treatment of autoimmune diseases. Background technique [0002] Autoimmune diseases occur when the body's immune system mistakenly attacks and destroys itself. There are currently as many as 80 autoimmune diseases, such as systemic lu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/365A61P37/02A61P19/02A61P3/10A61P1/04A61P1/00A61P1/16A61P17/00A61P29/00A61P17/06A61P21/04A61P21/00
CPCA61P1/00A61P1/04A61P1/16A61P3/10A61P17/00A61P17/06A61P19/02A61P21/00A61P21/04A61P29/00A61P37/02A61K31/365
Inventor 李婷刘良朱国元杨芬
Owner MACAU UNIV OF SCI & TECH
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