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Application of 4-methylumbelliferone in preparation of anti-allergic drugs

A methylumbelliferone and anti-allergic technology, which is applied in the field of biopharmaceuticals, can solve the problems of large side effects and poor anti-allergic drug effects, and achieve the effect of reducing degranulation and alleviating the symptoms of local and systemic allergic reactions in the body

Pending Publication Date: 2022-05-03
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of the above-mentioned deficiencies in the prior art, the object of the present invention is to provide a kind of application of 4-methylumbelliferone in the preparation of anti-allergic drugs, aiming to solve the problem that the existing anti-allergic drugs have relatively large side effects and poor anti-allergic drug effects. The problem

Method used

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  • Application of 4-methylumbelliferone in preparation of anti-allergic drugs
  • Application of 4-methylumbelliferone in preparation of anti-allergic drugs
  • Application of 4-methylumbelliferone in preparation of anti-allergic drugs

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Inhibitory effect of 4-MU on rat basophil (RBL-2H3) degranulation:

[0021] Mouse basophils are widely used in the diagnosis and immunotherapy of allergies. RBL-2H3 is easy to culture and is widely used in degranulation and signaling pathways, mast cell stabilizers, physicochemical properties of FcεR and its cytoskeleton Interaction studies etc. RBL-2H3 has high-affinity IgE receptors, which can be activated to secrete histamine and other transmitters by accumulating these receptors or synergizing with calcium ionophores.

[0022] In order to verify the inhibitory effect of 4-MU on the degranulation of rat basophils (RBL-2H3), its specific implementation is as follows:

[0023] Cell culture: Rat basophilic leukemia-2h3 cell lines (RBLs) were purchased from Cellcook Biotechnology Company (Guangzhou, China) and cultured in Dulbecco's Modified Eagle Medium (DMEM) containing 4.0 mM l-glutamine Amide, 1.0mM sodium pyruvate, 100U / ml penicillin, 100μg / ml streptomycin, non-es...

Embodiment 2

[0028] 4-MU inhibits degranulation of bone marrow-derived mast cells (BMMCs):

[0029] Cell culture: BMMCs were isolated from Balb / c mice (purchased from Guangdong Medical Experimental Animal Center), cultured in RPMI (Hyclone), 10% FBS (Gibico), 1% double antibody, 10ng / ml IL-3 and SCF4 -6 weeks.

[0030] CCK8 detects the toxicity of cells to 4-MU: BMMC (1×104 / well) were seeded in a 96-well plate, T-5224 was added after 24 hours, and incubation was continued for 24 hours. Then add 10ul CCK8 reagent, incubate for 1 hour, and finally measure the absorbance (OD value) with a microplate reader at a wavelength of 450nm.

[0031] Release of β-aminoglycosidase: Inoculate BMMC in a 24-well plate, incubate overnight with anti-DNP IgE for sensitization, incubate with 4-MU for 1 hour, stimulate with DNP-HSA (sigma) for 30 minutes, take 50ul supernatant and 50ul The substrate was reacted at 37°C for 90mins. The absorbance was measured at 405nm. After the supernatant was discarded, 500...

Embodiment 3

[0035] Effects of 4-MU on passive skin hypersensitivity in mice:

[0036] Breeding of BALB / c mice: Female mice (4-5 weeks old BALB / C) were purchased from the Guangdong Medical Laboratory Animal Center (Foshan, Guangdong) and placed in a temperature (24±1°C) and humidity (55±10 %) in a relatively stable SPF environment for 1 week. Mice were used to isolate bone marrow-derived mast cells (BMMC) and to perform a passive cutaneous anaphylaxis (PCA) model and an active systemic anaphylaxis (ASA) model.

[0037] Mice were injected with anti-DNP-IgE 500ng / ear. 24 hours later, 4-MU (50 mg / kg) or ketotifen (50 mg / kg, positive control group) was intraperitoneally injected (i.p.) into the ear (N=5 / group). One hour later, the mice were injected with 200 μg of DNP-HSA PBS containing 0.5% Evans blue dye into the tail vein. Mice were euthanized 1 h after DNP-HSA injection. Ears were taken and placed in 700 μl formamide for extraction at 65°C for 12 hours. Dye absorbance was measured at ...

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Abstract

The invention discloses an application of 4-methylumbelliferone in preparation of antiallergic drugs, the chemical structural formula of the 4-methylumbelliferone is shown in the specification, in an in-vitro experiment, through an in-vitro research method for comparing RBL-2H3 and BMMCs degranulation and an in-vivo experiment research method for a passive sensitization model PCA and an active systemic sensitization model ASA, the application of 4-methylumbelliferone in preparation of antiallergic drugs is realized, and the application of 4-methylumbelliferone in preparation of antiallergic drugs is realized. It is determined that the 4-methylumbelliferone can relieve the degranulation effect of IgE-mediated mast cells and effectively relieve local and systemic anaphylactic reaction symptoms of an organism.

Description

technical field [0001] The invention relates to the technical field of biopharmaceuticals, in particular to the application of 4-methylumbelliferone in the preparation of antiallergic drugs. Background technique [0002] Allergy is a state of hypersensitivity of the human immune system to substances in the normal environment. Common allergic diseases such as asthma, allergic rhinitis, and atopic dermatitis are all caused by allergic reactions. The symptoms are mainly redness and itching of the skin. , runny nose and shortness of breath, etc., bring continuous health threats to people's lives. In addition, about one-fifth of the people in the world suffer from allergic diseases, and social and economic development is related to the gradual increase in the incidence of allergic diseases. Allergic diseases have been listed by the World Health Organization (WHO) as the key prevention and treatment in the 21st century. It is one of the three major diseases in the world and is a ...

Claims

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Application Information

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IPC IPC(8): A61K31/366A61P37/08
CPCA61K31/366A61P37/08
Inventor 向秋安陈家杰乐湘柠王慧娜吉坤美
Owner SHENZHEN UNIV
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