And quercetin-3, 4apos, quercetin-3, 4apos; application of-diglucoside in preparation of oral ulcer treatment preparation and oral ulcer treatment preparation

Abstract

The invention provides application of quercetin 3, 4 '-diglucoside in preparation of a dental ulcer treatment preparation and the treatment preparation, and relates to the technical field of biological medicines. In the embodiment of the invention, based on genome-wide association study (GWAS) and Mendel randomization (MR) analysis, a series of patent medicine genes and related core targets of the oral ulcer are determined, and it is proved that quercetin-3, 4 '-diglucoside can be specifically combined with and regulate the core targets, so that accurate intervention on key pathological pathways of the oral ulcer is realized. The limitation that an existing oral ulcer treatment drug is unclear in target spot and fuzzy in action mechanism is overcome, a new natural product research and development paradigm driven by genetic evidence is provided, and a new drug candidate and solution are provided for oral ulcer treatment.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to the application of quercetin-3,4'-diglucoside in the preparation of oral ulcer treatment agents and the treatment agents thereof. Background Technology

[0002] Oral ulcers are one of the most common types of lesions on the oral mucosa, frequently occurring in conditions such as radiation stomatitis and infectious diseases. Recurrent aphthous stomatitis (RAS) is the most common oral mucosal disease, with a global incidence rate as high as 25%, of which more than 80% are mild cases. This disease is characterized by recurrent painful ulcers in the mouth, affecting patients' chewing, speech, and quality of life. Because the cause of RAS is unclear, there is currently no radical cure. Topical medications are the preferred treatment due to their good safety profile and significant pain relief. Given that over 80% of RAS patients have a mild form (a single small ulcer), their willingness to seek medical attention is generally low, and they tend to rely more on self-management measures. Therefore, incorporating natural active ingredients into daily oral care products is gradually becoming a convenient and effective treatment and management strategy.

[0003] However, the development of existing natural ingredient drugs largely relies on traditional experience and preliminary in vitro experiments, resulting in shortcomings such as unclear targets, ambiguous mechanisms of action, low development efficiency, and poor reproducibility of therapeutic effects. For example, existing oral care products containing natural ingredients are not precisely designed based on the core pathological pathways of the RAS, leading to a low match between active ingredients and disease targets, making it difficult to achieve highly effective treatment. Summary of the Invention

[0004] This invention provides the application of quercetin-3,4'-diglucoside in the preparation of oral ulcer treatment agents and the treatment agents themselves, in order to at least partially solve the problems existing in the prior art.

[0005] The first aspect of this invention provides the application of quercetin-3,4'-diglucoside in the preparation of oral ulcer treatment agents.

[0006] A second aspect of the present invention provides a treatment preparation for oral ulcers, the treatment preparation comprising: quercetin-3,4'-diglucoside and its derivatives.

[0007] Optionally, the therapeutic preparation is a spray, foam, powder, patch / film, gel / ointment, lozenge, or mouthwash.

[0008] Optionally, the therapeutic formulation may further include pharmaceutically acceptable excipients.

[0009] Optionally, pharmaceutically acceptable excipients may be any one or more of the following: thickeners, humectants, film-forming agents, diluents, disintegrants, lubricants, colorants, flavoring agents, binders, antioxidants, dispersants, preservatives, solvents, coagulants, pH adjusters and buffers, and penetration enhancers.

[0010] Optionally, the diluent is selected from one or more combinations of starch, pregelatinized starch, microcrystalline cellulose, powdered cellulose, dicalcium phosphate, lactose, sucrose, and sugar alcohol.

[0011] Optionally, the adhesive is selected from one or a combination of hydroxypropyl methylcellulose, hydroxypropyl cellulose, hydroxyethyl cellulose, polyvinylpyrrolidone, and sodium carboxymethyl cellulose.

[0012] Optionally, the lubricant is selected from magnesium stearate, sodium stearate fumarate, talc, and polyethylene glycol; the flow aid is selected from silica.

[0013] Optionally, the colorant is selected from one or a combination of several of water-soluble pigments, fat-soluble pigments, water-insoluble aluminum lakes, titanium dioxide, and iron oxides.

[0014] Optionally, the flavoring agent is selected from one or a combination of several of aspartame, acesulfame potassium, sucralose, steviol, menthol, citric acid, and vanillin.

[0015] Optionally, the pH adjuster and / or buffer is selected from one or a combination of several of the following: citric acid-sodium citrate, disodium hydrogen phosphate-sodium dihydrogen phosphate, sodium bicarbonate, sodium hydroxide, and hydrochloric acid.

[0016] Optionally, the penetration enhancer is selected from one or a combination of several of azone, oleic acid, polysorbate 80, bile salts, chitosan and its derivatives.

[0017] In this embodiment of the invention, based on genome-wide association studies (GWAS) and Mendelian randomization (MR), the core RAS target design allows quercetin-3,4'-diglucoside to precisely target the pathological pathways of oral ulcers, solving the problem of ambiguous drug targets in traditional drugs. In this embodiment of the invention, the active ingredient, quercetin-3,4'-diglucoside, is derived from a natural product, which can effectively avoid the side effects of drugs such as glucocorticoids and is suitable for daily use.

[0018] In this invention, a potential new use of quercetin-3,4'-diglucoside for the treatment of oral ulcers was discovered for the first time, establishing a genomics-guided paradigm for the development of natural ingredient drugs and providing a new strategy for the treatment of oral ulcers. Attached Figure Description

[0019] To more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the description of the embodiments of the present invention will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0020] Figure 1 The susceptible sites and differentially expressed genes supported by MR are shown in the embodiments of the present invention.

[0021] Figure 2 The results show that Mendelian randomization analysis supports a significant causal effect of NEK6 expression levels on oral ulcers.

[0022] Figure 3 The structural formula of quercetin-3,4'-diglucoside in an embodiment of the present invention is shown.

[0023] Figure 4 A structural model of the key component-target interaction is shown. Detailed Implementation

[0024] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, the present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0025] In the embodiments, for any experimental steps or conditions not specifically specified, the procedures and conditions described in conventional experimental procedures in the literature of this art can be followed. The genetic data involved in this invention are derived from European population genome data in publicly available databases (British Biobank and Finnish databases). Reagents and other instruments used, unless otherwise specified, are all commercially available products.

[0026] To overcome the technical problems existing in related technologies, this invention proposes to establish a precision R&D paradigm centered on disease mechanisms and driven by targets. This involves systematically analyzing the genetic susceptibility sites of RAS and the pathogenic pathways they regulate, identifying key druggable genes—clearly defining intervention targets from the genomic source; and then, guided by the targets, screening active molecules that can target and regulate the network through network pharmacology methods, providing a mechanistic basis for formulation design.

[0027] Specifically, genomic studies have revealed that the pathogenesis of RAS is closely related to immune regulation disorders and epithelial barrier damage. Genome-wide association studies (GWAS) and Mendelian randomization (MR) analyses can screen for key target genes of the disease (such as NEK6). Screening for targeted natural active ingredients based on these targets is a core approach to overcoming the shortcomings of current technologies.

[0028] To address the issues of unclear targets and limited treatment methods in existing oral ulcer treatments, this invention provides a RAS treatment drug with quercetin-3,4'-diglucoside as the core active ingredient, based on genomics-guided target screening, achieving safe, efficient, and convenient treatment of oral ulcers.

[0029] Quercetin, a flavonoid widely found in plants, possesses anti-inflammatory, immunomodulatory, and mucosal repair-promoting activities and has been used in the clinical treatment of oral ulcers. However, the targeted therapeutic potential of its glycosylated metabolites (such as quercetin-3,4'-diglucoside) has not yet been explored. For example, the application and mechanism of quercetin-3,4'-diglucoside in the treatment of recurrent oral ulcers (RAS) have not been reported.

[0030] To systematically identify genes associated with oral ulcers, this invention first conducted a cross-cohort genome-wide association study (GWAS) meta-analysis to identify genetic susceptibility loci. Then, causal genes were screened using summative Mendelian randomization (SMR) and multivariate Mendelian randomization (MVMR). Finally, differentially expressed genes (DEGs) were validated using transcriptome data from the Gene Expression Integrated Database (GEO) and the Genotype-Tissue Expression Database (GTEx). This meta-analysis identified 31 significant genome-wide loci associated with RAS risk (as shown in Table 1). KEGG pathway analysis revealed that susceptibility loci were significantly enriched in multiple immune-related pathways (e.g., Figure 1 As shown, Figure 1 (The results of KEGG pathway enrichment analysis of susceptible sites and differentially expressed genes (MR-DEGs) supported by MR are shown.) Table 1. Significant genome-wide loci associated with RAS identified by meta-analysis based on genome-wide association analysis (GWAS).

[0031] Based on the screened RAS genetic susceptibility loci, this invention performed SMR and MVMR analyses. These analyses identified significant quantitative trait loci (eQTLs) and protein quantitative trait loci (pQTLs) in blood and plasma. Subsequently, further integration with the RAS transcriptome dataset (GSE37265) yielded 265 high-confidence, MR-supported differentially expressed genes (including core targets such as NEK6). NEK6 is a serine / threonine kinase functionally associated with spindle assembly and chromosome segregation during mitosis (e.g., mitosis). Figure 2 As shown, Figure 2The results show that SMR-supported NEK6 expression levels have a significant causal effect on RAS, which is a key target for oral ulcers derived from the data of the embodiments of the present invention.

[0032] In this embodiment of the invention, drug repositioning analysis was performed on 265 key RAS targets across six drug databases, resulting in 74 candidate drugs. Since computational screening might identify drugs associated with both therapeutic effects and ulcer-like adverse reactions, this embodiment manually reviewed the list to determine its specific relevance to the pathogenesis or management of oral ulcers (as shown in Table 2). The validation results of this embodiment show that most candidate drugs are associated with oral mucositis as an adverse reaction. Notably, six of the 74 candidate drugs are recommended for the management of oral ulcers in clinical guidelines or authoritative literature, validating the reliability and predictive value of the research method in this embodiment.

[0033] Table 2. Targeted screening of medicinal plant components and repurposed drugs based on six major drug databases.

[0034] Furthermore, embodiments of this invention have identified a number of drugs with potential therapeutic potential but for which RAS indications have not yet been established, including five herbal ingredients with anti-inflammatory and antioxidant activities: capsaicin, papaverine, curcumin, quercetin, and retinol. Literature review has confirmed that capsaicin, curcumin, and quercetin have potential in the treatment of oral ulcers. Papaverine, however, was not considered for further research due to its potential adverse reactions.

[0035] Example 2: Validation of the mechanism of action (molecular docking and pathway analysis) By combining plant annotations for oral ulcers in HERB 2.0, a gene-ingredient-herb network was constructed. This network systematically links the genomic findings of this invention with empirical herbal treatments for oral ulcers. Quercetin is the most abundant bioactive ingredient, highlighting its potential as a core therapeutic agent for oral ulcers. However, quercetin is low in polarity and poor in water solubility. This poses a challenge in formulation development, often requiring specialized techniques to improve its solubility and bioavailability.

[0036] This invention proposes to attach a glucose molecule to each of the 3' and 4' carbon atoms of the quercetin molecule, thereby performing "glycosylation," such as... Figure 3 As shown, Figure 3The structural formula of quercetin-3,4'-diglucoside is shown. The hydrophilic nature of the linked glucose greatly enhances the compound's water solubility and bioavailability. This is a significant advantage for developing oral topical formulations such as mouthwashes, sprays, and toothpastes, as it allows for more efficient dissolution in aqueous matrices and better contact with the oral mucosa and ulcer surfaces. On one hand, quercetin-3,4'-diglucoside, as a prodrug, can be hydrolyzed in vivo by β-glucosidase, releasing high concentrations of quercetin locally at target sites (such as inflamed tissues, areas with high levels of certain bacteria or cellular enzymes), thereby exerting its therapeutic effect. On the other hand, quercetin-3,4'-diglucoside itself can directly bind to specific targets as an active molecule, exerting quercetin-like anti-inflammatory and antioxidant effects. Specific data are as follows:

[0037] Molecular docking: Molecular docking was performed using AutoDock software. The results showed that quercetin or quercetin-3,4'-diglucoside could directly bind to NEK6 (e.g., Figure 4 As shown, Figure 4 A structural model of the key component-target interaction is shown. Specifically, quercetin forms hydrogen bonds with only a few amino acid residues of NEK6 (TRP-201, MET-203, and SER-234), with bond lengths ranging from 2.3 to 3.2 Å. In contrast, quercetin-3,4'-diglucoside binds more extensively to NEK6, interacting not only with the aforementioned residues but also with residues such as ASN-29, ASP-84, and TYR-98, forming a denser hydrogen bond network (bond lengths ranging from 2.5 to 3.4 Å). Binding energy scores further support this result: the binding score of quercetin-3,4'-diglucoside is -10.023, superior to quercetin's -9.247, suggesting a stronger theoretical affinity for NEK6. Given that quercetin has been clinically proven to be effective in treating oral ulcers, this study reveals at the molecular level that its glycosylated derivatives may have greater potential in treating oral ulcers by enhancing their binding to NEK6, providing evidence for its mechanism of action.

[0038] In this embodiment of the invention, based on the RAS core target design screened by GWAS and MR analysis, quercetin-3,4'-diglucoside precisely acts on the key target NEK6 of oral ulcers, solving the problem of ambiguous target of traditional drugs. In this embodiment of the invention, the active ingredient, quercetin-3,4'-diglucoside, is derived from a natural product. No adverse events have been reported in clinical trials, thus avoiding the side effects of drugs such as glucocorticoids and making it suitable for long-term daily use.

[0039] In this invention, a potential new use of quercetin-3,4'-diglucoside for the treatment of oral ulcers was discovered for the first time, and the therapeutic mechanism of quercetin-3,4'-diglucoside for oral ulcers was clarified. A genomics-guided paradigm for the development of natural ingredient drugs was established, providing a new strategy for the treatment of RAS.

[0040] Based on the exploratory research of the above embodiments, the present invention specifically proposes the application of quercetin-3,4'-diglucoside in the preparation of oral ulcer treatment agents.

[0041] Based on the exploratory research of the above embodiments, the present invention also proposes a treatment preparation for oral ulcers, the treatment preparation comprising: quercetin-3,4'-diglucoside and its derivatives.

[0042] Specifically, the therapeutic preparation is a spray, foam, powder, patch / film, gel / ointment, lozenge, or mouthwash.

[0043] Specifically, the therapeutic formulation also includes pharmaceutically acceptable excipients.

[0044] Specifically, pharmaceutically acceptable excipients include any one or more of the following: thickeners, humectants, film-forming agents, diluents, disintegrants, lubricants, colorants, flavoring agents, binders, antioxidants, dispersants, preservatives, solvents, coagulants, pH adjusters and buffers, and penetration enhancers.

[0045] Specifically, the diluent is selected from one or more combinations of starch, pregelatinized starch, microcrystalline cellulose, powdered cellulose, dicalcium phosphate, lactose, sucrose, and sugar alcohol.

[0046] Specifically, the adhesive is selected from one or more combinations of hydroxypropyl methylcellulose, hydroxypropyl cellulose, hydroxyethyl cellulose, polyvinylpyrrolidone, and sodium carboxymethyl cellulose.

[0047] Specifically, the lubricant is selected from magnesium stearate, sodium stearate fumarate, talc, and polyethylene glycol; the flow aid is selected from silicon dioxide.

[0048] Specifically, the colorant is selected from one or a combination of several of the following: water-soluble pigments, fat-soluble pigments, water-insoluble aluminum lakes, titanium dioxide, and iron oxides.

[0049] Specifically, the flavoring agent is selected from one or more of the following: aspartame, acesulfame potassium, sucralose, steviol glycosides, menthol, citric acid, and vanillin.

[0050] Specifically, the pH adjuster and / or buffer is selected from one or more combinations of citric acid-sodium citrate, disodium hydrogen phosphate-sodium dihydrogen phosphate, sodium bicarbonate, sodium hydroxide, and hydrochloric acid.

[0051] Specifically, the penetration enhancer is selected from one or more combinations of azone, oleic acid, polysorbate 80, bile salts, chitosan and its derivatives.

[0052] Although preferred embodiments of the present invention have been described, those skilled in the art, upon learning the basic inventive concept, can make other changes and modifications to these embodiments. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments as well as all changes and modifications falling within the scope of the embodiments of the present invention.

[0053] The application and therapeutic formulation of quercetin-3,4'-diglucoside provided by this invention in the preparation of oral ulcer treatment preparations have been described in detail above. Specific examples have been used to illustrate the principles and implementation methods of this invention. The above description of the embodiments is only for the purpose of helping to understand the method and core ideas of this invention. At the same time, for those skilled in the art, there will be changes in the specific implementation methods and application scope based on the ideas of this invention. Therefore, the content of this specification should not be construed as a limitation of this invention.

Claims

1. Application of quercetin-3,4'-diglucoside in the preparation of oral ulcer treatment agents.

2. A treatment preparation for oral ulcers, characterized in that, The therapeutic agents include: quercetin-3,4'-diglucoside and its derivatives.

3. The oral ulcer treatment preparation according to claim 2, characterized in that, The therapeutic preparation is a spray, foam, powder, patch / film, gel / ointment, lozenge, or mouthwash.

4. The oral ulcer treatment preparation according to claim 2, characterized in that, The therapeutic formulation also includes pharmaceutically acceptable excipients.

5. The oral ulcer treatment preparation according to claim 4, characterized in that, Pharmaceutically acceptable excipients include any one or more of the following: thickeners, humectants, film-forming agents, diluents, disintegrants, lubricants, colorants, flavoring agents, binders, antioxidants, dispersants, preservatives, solvents, coagulants, pH adjusters and buffers, and penetration enhancers.

6. The oral ulcer treatment preparation according to claim 5, characterized in that, The diluent is selected from one or more combinations of starch, pregelatinized starch, microcrystalline cellulose, powdered cellulose, dicalcium phosphate, lactose, sucrose, and sugar alcohol; The adhesive is selected from one or a combination of several of hydroxypropyl methylcellulose, hydroxypropyl cellulose, hydroxyethyl cellulose, polyvinylpyrrolidone, and sodium carboxymethyl cellulose.

7. The oral ulcer treatment preparation according to claim 5, characterized in that, The lubricant is selected from magnesium stearate, sodium stearate fumarate, talc, and polyethylene glycol; the flow aid is selected from silica. The colorant is selected from one or a combination of several of the following: water-soluble pigments, fat-soluble pigments, water-insoluble aluminum lakes, titanium dioxide, and iron oxides.

8. The oral ulcer treatment preparation according to claim 5, characterized in that, The flavoring agent is selected from one or more of aspartame, acesulfame potassium, sucralose, steviol glycoside, menthol, citric acid, and vanillin.

9. The oral ulcer treatment preparation according to claim 5, characterized in that, pH adjusters and / or buffers are selected from one or more of the following: citric acid-sodium citrate, disodium hydrogen phosphate-sodium dihydrogen phosphate, sodium bicarbonate, sodium hydroxide, and hydrochloric acid.

10. The oral ulcer treatment preparation according to claim 5, characterized in that, The penetration enhancer is selected from one or a combination of several of azone, oleic acid, polysorbate 80, bile salts, chitosan and its derivatives.

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