Application of chenodeoxycholic acid in preparation of medicine for treating allergic diseases

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to application of chenodeoxycholic acid in preparation of drugs for treating allergic diseases. It is found that CDCA can effectively inhibit pyroptosis and secretion of proinflammatory factors, expression of complement inhibition proteins CD46, CD55 and CD59 is remarkably up-regulated, expression of the complement inhibition proteins CD46, CD55 and CD59 is finally promoted, and occurrence and development of allergic diseases are effectively relieved. In a mouse model, CDCA can significantly improve OVA-induced food allergy symptoms, and has application prospects in treatment of allergic diseases.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to the application of chenodeoxycholic acid in the preparation of drugs for treating allergic diseases. Background Technology

[0002] Allergic diseases are a series of illnesses caused by an abnormal immune response of the human immune system to certain foreign substances (allergens). Under normal circumstances, the immune system can recognize and defend against harmful substances. However, in an allergic reaction, the immune system mistakenly attacks harmless substances (such as pollen and dust mites), triggering a series of inflammatory responses. The main mechanism involves the activation of the complement system. During complement system activation, various protein fragments, such as C3a, C4a, and C5a, are produced. These anaphylatoxins can bind to corresponding receptors on the cell membrane surface and mediate various biological functions, such as causing degranulation of mast cells and basophils, releasing inflammatory mediators like histamine, increasing permeability and allowing fluid to seep into the skin tissue, resulting in symptoms such as erythema, papules, and itching, thus contributing to the occurrence of allergic reactions. Furthermore, anaphylatoxins C3a and C5a can chemotactically activate neutrophils and macrophages, causing these inflammatory cells to accumulate at the site of the allergic reaction, further exacerbating the inflammation. Inhibiting the activation of the complement system or blocking the effects of anaphylatoxins may become a new strategy for treating allergic diseases.

[0003] Complement inhibitory proteins are a class of proteins that regulate the activity of the complement system, playing a crucial role in maintaining immune system homeostasis and preventing tissue damage caused by excessive complement activation. CD46 (membrane cofactor protein, MCP) is a cofactor of complement factor I, promoting the cleavage of C3b and C4b by factor I, thereby inhibiting C3 convertase activity and preventing excessive activation of the complement system. CD55 (decay accelerator factor, DAF) accelerates the decay of C3 convertases (C3bBb and C4b2a), preventing further activation of the complement system. CD59 (membrane reactive lysis inhibitor, MRL) prevents the assembly of the membrane attack complex (MAC), preventing C9 insertion and polymerization on the cell membrane, thus protecting cells from MAC-mediated lysis. These three important membrane-bound complement regulatory proteins protect host cells from complement-mediated damage. Currently, the specific mechanisms of allergic diseases are unclear; therefore, it is urgent to elucidate the pathological mechanisms of allergic diseases and to develop new strategies targeting the complement pathway for the prevention and treatment of allergic diseases.

[0004] Chenodeoxycholic acid (CDCA) is a bile acid primarily used to treat cholesterol gallstones and plays a regulatory role in cholestatic diseases. Current technology has not yet found its application in treating allergic diseases. Summary of the Invention

[0005] The purpose of this invention is to provide the use of chenodeoxycholic acid or its derivatives in the preparation of products for treating allergic diseases.

[0006] To achieve the above-mentioned objectives of this invention, the technical solution adopted by this invention is as follows: This invention provides the use of chenodeoxycholic acid or its derivatives in the preparation of medicaments for treating allergic diseases.

[0007] In some embodiments of the present invention, the chenodeoxycholic acid or its derivatives include one of chenodeoxycholic acid, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable modification thereof, an isomer thereof, or a secondary metabolite thereof.

[0008] In some embodiments of the present invention, the isomers include ursodeoxycholic acid.

[0009] The chemical formula for chenodeoxycholic acid (CDCA) is C2. 24 H 40 O4, CAS number 474-25-9, has an α-configuration (axial, extending below the plane of the steroid ring) at the 7th hydroxyl position, and its chemical name is 3α,7α-dihydroxy-5β-cholestane-24-acid.

[0010] Ursodeoxycholic acid (UDCA) has the chemical formula C0. 24 H 40 O4, CAS number 128-13-2, has a β-configuration (equatorial bond, parallel to the plane of the steroid ring) at the 7th hydroxyl position, and its chemical name is 3α,7β-dihydroxy-5β-cholestane-24-acid.

[0011] In some embodiments of the invention, the pharmaceutically acceptable salt includes an alkali addition salt.

[0012] In some embodiments of the present invention, a "pharmaceutically acceptable base addition salt" refers to a salt that retains the biological effectiveness and properties of the free acid and is not undesirable in biological or other respects. These salts are prepared by the addition of an inorganic or organic base to a free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts. Preferred inorganic salts are ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, the following: primary, secondary, and tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines, and basic ion exchange resins such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, ethylenediamine, glucosamine, methylglucosamine, theobromine, triethanolamine, thiocyanate, purine, piperazine, piperidine, N-acetylene, etc. Ethylpiperidine, polyamine resins, etc. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine.

[0013] In some embodiments of the present invention, the pharmaceutically acceptable modification includes at least one of phosphorylation, sulfonation, acylation, glycosylation, ubiquitination, acetylation, methylation, sulfation, phospholipidation, and halogenation.

[0014] In some embodiments of the present invention, the secondary metabolites include at least one of deoxycholic acid, lithocholic acid, and 7-ketolithocholic acid.

[0015] In some embodiments of the present invention, the allergic disease includes at least one of allergic rhinitis, allergic asthma, allergic gastroenteritis, food allergy, allergic conjunctivitis, eczema, urticaria, and contact dermatitis; preferably, it is food allergy.

[0016] In some embodiments of the present invention, the symptoms of the allergic disease include at least one of the following: nasal itching, sneezing (usually multiple in succession), runny nose, nasal congestion, itchy eyes, tearing, red eyes, swollen eyelids, wheezing, shortness of breath, chest tightness, cough, rash, itching, urticaria, purpura, abdominal pain, diarrhea, vomiting, constipation, wheezing, difficulty breathing, and laryngeal edema.

[0017] In some embodiments of the present invention, chenodeoxycholic acid or its derivatives can effectively inhibit pyroptosis and the secretion of pro-inflammatory factors, activate the complement pathway, significantly upregulate the expression of complement inhibitory proteins CD46 / 55 / 59, and ultimately promote the expression of complement inhibitory proteins CD46 / 55 / 59, effectively alleviating the occurrence and progression of allergic diseases.

[0018] In some embodiments of the present invention, the complement pathway activation includes at least one of the following: aberrant activation of the classical complement pathway, the alternative pathway, the lectin pathway, and the granzyme K pathway.

[0019] In some embodiments of the present invention, the activation of the complement pathway is manifested as: abnormal expression of at least one of the complement proteins, including C2a, C4b, C3, and C3b, and complement inhibitory proteins CD46 / 55 / 59 in the classical complement pathway, the alternative pathway, the lectin pathway, and the granzyme K pathway.

[0020] In some embodiments of the present invention, the pyroptosis includes at least one of the expression changes of the GSDM family members GSDMA, GSDMB, GSMDC, GSDMD, and GSDME.

[0021] In some embodiments of the present invention, the pro-inflammatory factor includes at least one of the pyroptosis products IL1b and IL18.

[0022] In some embodiments of the invention, the product includes pharmaceutically acceptable excipients.

[0023] In some embodiments of the present invention, the pharmaceutically acceptable excipients include at least one of the following: propellants, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, flow aids, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesion agents, penetration enhancers, pH adjusters, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, encapsulating agents, humectants, absorbents, diluents, flocculants and anti-flocculators, filter aids, and release inhibitors.

[0024] The pharmaceutically acceptable excipients mentioned above are generally recognized for use in this purpose and as inactive ingredients in the pharmaceutical preparation. Compilations of pharmaceutically acceptable excipients can be found in reference books such as the *Handbook of Pharmaceutical Excipients* (2nd edition, edited by A. Wade and PJ Weller; published by the American Pharmaceutical Association, Washington and The Pharmaceutical 6Gess, London, 1994) and the *Pharmacopoeia of the People's Republic of China - List of Pharmaceutical Excipients*.

[0025] In some embodiments of the present invention, the dosage form of the product includes one of the following: powder, tablet, granule, capsule, sustained-release agent, solution, dry suspension, effervescent tablet, emulsion, suspension, syrup, drops, and chewable tablet.

[0026] In some embodiments of the present invention, the product is administered via the gastrointestinal tract or non-gastrointestinal route.

[0027] In some embodiments of the present invention, the gastrointestinal administration includes one of oral administration, sublingual administration, and rectal administration.

[0028] In some embodiments of the present invention, the non-gastrointestinal administration includes one of intravenous injection, subcutaneous injection, and mucosal administration.

[0029] In some embodiments of the present invention, the product is applied to mammals, including but not limited to: humans, mice, rats, pigs, cattle, sheep, horses, monkeys, and rabbits.

[0030] In some embodiments of the invention, the mammal includes humans.

[0031] In some embodiments of the present invention, the drug further includes any one or more other active ingredients; the other active ingredients have the function of treating allergies.

[0032] The beneficial effects of this invention are: This invention reveals that CDCA can effectively inhibit pyroptosis and the secretion of pro-inflammatory factors, and significantly upregulate the expression of complement inhibitory proteins CD46, CD55, and CD59, ultimately promoting the expression of these proteins and effectively alleviating the progression of allergic diseases. In a mouse model, CDCA can significantly improve OVA-induced food allergy symptoms, demonstrating promising applications in the treatment of allergic diseases. Attached Figure Description

[0033] The present invention will be further described below with reference to the accompanying drawings and embodiments, wherein: Figure 1 To illustrate the use of CDCA to alleviate OVA-induced food allergy symptoms in this invention: (A) Animal experimental group treatment model; (B) Itching index score; (C) Liver weight difference analysis; (D) Spleen weight difference; (E) HE staining; (F) Immunofluorescence staining showing IgE; (G) Immunofluorescence staining showing mast cell activation.

[0034] Figure 2In this invention, CDCA was used to alleviate the reduction of complement inhibitor protein induced by OVA; (A) Western blot was used to detect pyroptosis and expression of complement inhibitor protein in small intestinal tissue; (B) Immunofluorescence assay was used to detect the expression of complement inhibitor protein in small intestinal tissue sections; (C) Immunohistochemistry was used to show the expression of IFNr in the spleen. Detailed Implementation

[0035] The following will describe the concept and technical effects of the present invention clearly and completely with reference to embodiments, so as to fully understand the purpose, features and effects of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are all within the scope of protection of the present invention.

[0036] The animal experiments related to this invention have been approved by the Animal Ethics Review Committee of the Women and Children's Medical Center Affiliated to Guangzhou Medical University.

[0037] Example 1: CDCA alleviates OVA-induced food allergy 1. Experimental Methods Experimental animals: C57BL6 mice.

[0038] Experimental reagents: feed containing 0.2 wt% chenodeoxycholic acid.

[0039] Experimental groups: control group; allergy group (OVA-induced allergy model); treatment group (OVA allergy model + CDCA treatment group), randomly divided into 3 groups, with 5 animals in each group.

[0040] Experimental Procedure: The control group was fed a standard diet as usual. In the allergy group, during the sensitization phase, each animal was injected intraperitoneally with 100 μL of feed containing 100 μg OVA and 1 mg Al(OH)3, three times consecutively, with a one-week interval between each injection. Subsequently, after a two-week interval, each animal was administered a high dose of 100 mg OVA via gavage for five consecutive times. In the treatment group, after the sensitization phase, the animal was switched to a diet containing chenodeoxycholic acid.

[0041] Throughout the process, anaphylactic symptoms were scored (anaphylactic symptoms scoring criteria: no symptoms = 0, frequent scratching of ears and nose = 1, red rashes or ulcers in the mouth and tail = 3, spasms or muscle contractions = 4, shock or death within 30 minutes = 5), and liver and spleen weight differences were recorded. Subsequently, tissue samples were taken for relevant pathological examinations to detect complement inhibitory proteins CD46, CD55, and CD59 in the small intestine and IFNr expression in the spleen.

[0042] 2. Experimental Results Experimental results are as follows Figure 1As shown, (A) is a schematic diagram of the experimental groups and treatments, namely the control group, the OVA group (allergy group), and the OVA+CDCA group (treatment group). (B) CDCA feeding significantly reduced the allergic reaction symptom score. (C) CDCA feeding significantly reduced hepatomegaly and liver weight in the allergy group. (D) CDCA feeding had no significant effect on spleen weight in the allergy group. (E) HE staining results showed that CDCA feeding significantly reduced intestinal inflammation in the small intestine tissue of the allergy group. (F) Immunofluorescence results showed that CDCA reduced the expression level of IgE in the small intestine tissue of the allergy group. (G) Immunofluorescence results showed that CDCA reduced the number and activation of mast cells in the small intestine tissue of the allergy group.

[0043] The above results indicate that CDCA can effectively alleviate food allergy-related symptoms and intestinal inflammation.

[0044] Example 2: CDCA alleviates OVA-induced reduction in complement inhibitory proteins. 1. Experimental Methods Small intestines were taken from the experimental animals in Example 1 for Western blot analysis of complement inhibitory proteins CD46 / 59, Caspase 1, GSDMB, GSDMC, GSDMD, GSDME, and IL-18; small intestines were taken for immunofluorescence analysis of the expression of CD46, CD55, and CD59; spleens were taken for immunohistochemical staining with IFNgamma.

[0045] 2. Experimental Results Experimental results are as follows Figure 2 As shown, (A) Western blot results from small intestinal tissue showed that complement inhibitory proteins CD46 and CD59 were significantly decreased in the allergy group, while the expression of pyroptosis-related proteins N-GSDM family proteins and pro-inflammatory factor IL18 was significantly increased in the allergy group. After OVA treatment, the expression of complement inhibitory proteins CD46 and CD59 was significantly increased, and the expression of N-GSDM family proteins and pro-inflammatory factor IL18 was downregulated. (B) Immunofluorescence results showed that complement inhibitory proteins CD46, CD55, and CD59 were highly expressed in the small intestinal epithelial cells. The expression of complement inhibitory proteins CD46, CD55, and CD59 was significantly decreased in the allergy group, while the expression of complement inhibitory proteins CD46, CD55, and CD59 in the small intestinal epithelial cells was increased in the CDCA-fed treatment group. (C) Immunohistochemical staining showed that, compared with the control group, the expression of IFNgamma was significantly increased in the spleen tissue of the allergy group, while the expression of IFNgamma was downregulated in the CDCA-fed treatment group.

[0046] The above results indicate that CDCA can alleviate food allergies by upregulating complement inhibitory proteins, inhibiting pyroptosis, and suppressing the expression of inflammatory factors.

[0047] The embodiments of the present invention have been described in detail above with reference to the accompanying drawings. However, the present invention is not limited to the above embodiments, and various changes can be made within the scope of knowledge possessed by those skilled in the art without departing from the spirit of the present invention. Furthermore, the embodiments of the present invention and the features thereof can be combined with each other unless otherwise specified.

Claims

1. The use of chenodeoxycholic acid or its derivatives in the preparation of drugs for treating allergic diseases.

2. The application according to claim 1, characterized in that: The chenodeoxycholic acid or its derivatives include one of chenodeoxycholic acid, its pharmaceutically acceptable salt, pharmaceutically acceptable modification, isomer, or secondary metabolite.

3. The application according to claim 2, characterized in that: The isomers include ursodeoxycholic acid.

4. The application according to claim 2, characterized in that: The secondary metabolites include at least one of deoxycholic acid, lithocholic acid, and 7-ketolithocholic acid.

5. The application according to claim 1, characterized in that: The allergic diseases include at least one of allergic rhinitis, allergic asthma, allergic gastroenteritis, food allergy, allergic conjunctivitis, eczema, urticaria, and contact dermatitis.

6. The application according to claim 1, characterized in that: The drug includes pharmaceutically acceptable excipients.

7. The application according to claim 6, characterized in that: The pharmaceutically acceptable excipients include at least one of the following: propellants, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, flow aids, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesion agents, penetration enhancers, pH adjusters, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, encapsulating agents, humectants, absorbents, diluents, flocculants and anti-flocculation agents, filter aids, and release inhibitors.

8. The application according to claim 1, characterized in that: The dosage forms of the drug include those administered via the gastrointestinal tract or those administered outside the gastrointestinal tract.

9. The application according to claim 1, characterized in that: The product is intended for use on mammals; Preferably, the mammal includes humans.

10. The application according to claim 1, characterized in that: The drug also includes any one or more other active ingredients; The other active ingredients have the function of treating allergies.

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