A composition of arctiin and acacia and its application

Abstract

The application discloses a composition of arctiin and farnesol and application thereof and belongs to the technical field of medicines. The application provides a composition of arctiin and farnesol, which comprises arctiin and farnesol. The combination of arctiin and farnesol can achieve good synergistic effect, can well inhibit the release activity of IL-6, IL-1beta and TNF-alpha, can affect the expression of key proteins of apoptosis, oxidative stress and anti-inflammatory pathways, can significantly improve the anti-inflammatory effect, and has simple components, clear mechanism and improved medication safety of anti-inflammatory drugs.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a composition of arctiin and farnesin and its application. Background Technology

[0002] Inflammation is a defensive response of living tissue with a vascular system to damaging factors. It is a defensive reaction of the body to stimuli, manifested as redness, swelling, heat, pain, and functional impairment. Inflammation can be infectious inflammation caused by infection or non-infectious inflammation not caused by infection. During the inflammatory process, on the one hand, damaging factors directly or indirectly cause damage to tissues and cells; on the other hand, inflammatory hyperemia and exudation dilute, kill, and surround the damaging factors. Simultaneously, the regeneration of parenchymal and interstitial cells allows damaged tissues to be repaired and healed. Therefore, inflammation can be said to be a unified process of damage and anti-damage. The main cellular natural host defense mechanisms are the phagocytosis and killing of bacteria and fungi by neutrophils, monocytes, and macrophages, and the lysis of virus-infected cells by natural killer (NK) cells. Once microorganisms are recognized, pro-inflammatory cytokines are secreted. These cytokines activate neutrophils and macrophages to phagocytose invading pathogens and release toxic oxygen and nitrogen free radicals. Among the many inflammatory cytokines, the main ones are TNF-α (tumor necrosis factor-α), IL-1β (interleukin-1β), and IL-6 (interleukin-6).

[0003] TNF-α is the earliest and most important inflammatory mediator in the inflammatory response. It activates neutrophils and lymphocytes, increases vascular endothelial cell permeability, regulates metabolic activity in other tissues, and promotes the synthesis and release of other cytokines. It is mainly produced by macrophages, with LPS being a strong stimulant. T cells and NK cells can also secrete TNF-α under the influence of certain stimulating factors (such as PMA). IL-1β is secreted through unconventional protein secretion pathways, and can be paracrine or systemic. It is mainly produced by sentinel cells of the innate immune system: monocytes and macrophages, and can also be released from epithelial cells, endothelial cells, fibroblasts, synovial cells, neurons, mast cells, and glial cells such as microglia and astrocytes. IL-6 is mainly produced by macrophages, T cells, B cells, and other cells. It is a multifunctional cytokine that regulates immune responses, hematopoiesis, acute phase responses to tissue damage, and inflammatory responses. IL-6 levels are elevated in cardiovascular and cerebrovascular diseases, kidney diseases, tumors, arthritis, trauma, and infections. IL-6 is closely related to respiratory diseases, kidney diseases, tumors, and other diseases.

[0004] Arctiin (ARC) is a lignan compound extracted and isolated from the dried, mature fruit of *Arctium lappa*, a biennial herbaceous plant belonging to the genus *Arctium* of the Asteraceae family. A small amount is also found in burdock leaves. Burdock fruit is a traditional Chinese medicine, primarily used to treat wind-heat colds, measles, carbuncles, and boils. Modern pharmacology has demonstrated that burdock fruit also possesses multiple effects, including antibacterial, antitumor, and hypoglycemic properties. In vivo and in vitro pharmacokinetic studies have shown that ARC produces at least two metabolites in the gastrointestinal tract: ARC-G (M1) and 2-(3″,4″-dihydroxybenzyl)-3-(3′,4′-dimethoxybenzyl)butyrolactone (M2). After intestinal absorption, M2 is methylated into M1 by COMT in the liver. Thus, only the M1 form, ARC-G, exists in the blood. ARC-G is then transported to various organs by the bloodstream to exert its effects.

[0005] Acacetin is a natural flavonoid compound widely found in nature, such as in acacia trees, locust trees, thistles, clematis, buddleja officinalis, and chrysanthemums. Acacetin is also known as 5,7-dihydroxy-4'-methoxyflavonoid, robinin, and robinin, and its molecular formula is Ca. 16 H 12 O5 has a molecular weight of 284.27. In addition to its traditional effects of relieving depression, calming the mind, regulating qi, and promoting blood circulation, acacia extract also has a variety of biological activities, such as antioxidant, anti-inflammatory, antibacterial, and antiviral properties, and can treat many tumor diseases through multiple pathways.

[0006] Since there is a need to further develop drugs that inhibit the expression of inflammatory cytokines and inflammatory mediators, and although arctiin and farnesin have certain anti-inflammatory effects, their efficacy still needs to be improved. In this study, we found that the combined use of arctiin and farnesin has a synergistic effect. Summary of the Invention

[0007] The purpose of this invention is to provide a composition of arctiin and farnesin and its application. During the experiment, the applicant unexpectedly discovered that when arctiin and farnesin are used in combination, they can achieve a synergistic anti-inflammatory effect.

[0008] To achieve the above-mentioned objectives, the technical solution of this invention is as follows:

[0009] On one hand, the present invention provides a composition of arctiin and farnesin, comprising arctiin and farnesin.

[0010] Preferably, the mass ratio of arctiin to farnesin is 2-8:2-8.

[0011] Further preferred, the mass ratio of arctiin to farnesin is 1:1.

[0012] On the other hand, the present invention relates to the application of the above-mentioned arctiin and farnesin composition in the preparation of anti-inflammatory drugs.

[0013] Preferably, the anti-inflammatory drug exerts its anti-inflammatory effect by inhibiting the expression of inflammatory factors. Therefore, diseases related to the overexpression of inflammatory factors, including but not limited to rheumatoid arthritis, psoriasis, asthma, atopic dermatitis, allergic rhinitis, prostatitis, inflammatory bowel disease, and diabetes, can all be treated with the compositions of the present invention.

[0014] Preferably, the inflammatory factor is one or more selected from interleukin, tumor necrosis factor α, transforming growth factor β, bradykinin, oxygen free radical, and nitric oxide. More preferably, it is interleukin or tumor necrosis factor α.

[0015] Preferably, the interleukin is one or more of IL1, IL4, and IL6. More preferably, the interleukin is IL6.

[0016] An anti-inflammatory drug comprising the above-mentioned combination of arctiin and farnesin and pharmaceutically acceptable excipients.

[0017] The drug can be prepared into dosage forms such as pills, capsules, granules, oral liquids, powders, tablets, lozenges, and sugar lozenges. For different dosage forms, a suitable drug carrier in this field can be selected.

[0018] The drug carrier used can be solid, liquid, or gaseous. Examples of solid carriers include lactose, kaolin, sucrose, talc, gelatin, agar, pectin, gum arabic, magnesium stearate, and stearic acid. Examples of liquid carriers include syrup, peanut oil, olive oil, and water. Examples of gaseous carriers include carbon dioxide and nitrogen.

[0019] In preparing compositions for oral dosage forms, any convenient pharmaceutical medium can be used. For example, water, ethanol, oils, alcohols, flavoring agents, preservatives, coloring agents, etc., can be used to form oral liquid dosage forms, such as suspensions, solutions, and granules; while carriers, such as starch, sugars, microcrystalline cellulose, diluents, granulators, emulsifiers, lubricants, binders, and disintegrants, can be used to form oral solid dosage forms, such as powders, capsules, and tablets. Tablets and capsules are preferred oral dosage units using solid pharmaceutical carriers due to their ease of administration. Tablets can be coated using standard aqueous or non-aqueous techniques.

[0020] Tablets containing the traditional Chinese medicine composition of the present invention can be prepared by compression or molding, and one or more excipients or adjuvants may be used. The active ingredient can be compressed in a free-flowing form (e.g., powder or granules) in a suitable machine, optionally mixed with binders, lubricants, inert diluents, surfactants, or dispersants. Molded tablets can be molded in a suitable machine, i.e., a mixture of powdered compounds moistened with an inert liquid diluent. Each tablet preferably contains about 0.05 mg to about 5 g of active ingredient, and each sachet or capsule preferably contains about 0.05 mg to about 5 g of active ingredient. For example, a formulation intended for oral administration to humans may contain about 0.5 mg to about 5 g of active drug, mixed with a suitable and convenient carrier material, which may comprise about 5% to 95% of the total composition. Unit dosage forms typically contain about 1 mg to about 2 g of active ingredient, typically in doses of 25 mg, 50 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg, or 1000 mg.

[0021] The pharmaceutical compositions of this invention suitable for parenteral administration can be prepared as aqueous solutions or suspensions of the active compound. Suitable surfactants, such as hydroxypropyl cellulose, may be included. Dispersions can also be prepared in mixtures of glycerol, liquid polyethylene glycol, and their oils. Furthermore, preservatives may be added to prevent harmful microbial growth.

[0022] The pharmaceutical products suitable for injection in this invention include sterile aqueous solutions or dispersions. Furthermore, the pharmaceutical product can be in the form of a sterile powder for the ad hoc preparation of such sterile injection solutions or dispersions. In all cases, the final injectable form must be sterile and must be an effective liquid so that the injectable pharmaceutical ingredient must remain stable under the production and storage conditions; therefore, it is best to preserve it to prevent contamination by microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium, such as containing water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), vegetable oils, and suitable mixtures thereof.

[0023] The medicaments of the present invention can be in forms suitable for topical use, such as aerosols, creams, ointments, lotions, powders, or the like. Furthermore, the compositions can be in suitable forms for transdermal drug delivery devices. These formulations can be prepared using the traditional Chinese medicine compositions of the present invention through conventional processing methods. For example, a cream or ointment with a desired consistency can be prepared by mixing a hydrophilic material and water, and about 5 wt% to about 10 wt% of the compound.

[0024] The medicament of the present invention can be in a form suitable for rectal administration, wherein the carrier is solid. It is preferable to formulate the mixture into a single-dose suppository. Suitable carriers include cocoa butter and other materials commonly used in the art. Suppositories can be made by first forming a mixture containing a softened or melted carrier, followed by cooling and shaping in a mold.

[0025] In addition to the carrier components described above, the pharmaceutical formulations may include (if applicable) one or more additional carrier components, such as diluents, buffers, flavoring agents, binders, surfactants, thickeners, lubricants, preservatives (including antioxidants), etc. Furthermore, other excipients may be added, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, etc., colorants, and flavoring agents, etc., to make the formulation isotonic with the blood of the intended receptor. Components containing the traditional Chinese medicine composition of this invention can also be prepared in powder or concentrated form.

[0026] The beneficial effects of this invention are as follows:

[0027] This invention provides a pharmaceutical composition that combines arctiin and farnesin to achieve a good synergistic effect. It can effectively inhibit the release activity of IL-6, IL-1β, and TNF-α, affect the expression of key proteins in apoptosis, oxidative stress, and anti-inflammatory pathways, and significantly improve the anti-inflammatory effect. Moreover, the composition is simple and the mechanism is clear, which improves the safety of anti-inflammatory drugs. Attached Figure Description

[0028] Figure 1 This is a graph showing the results of IL-1β secretion.

[0029] Figure 2 This is a graph showing the results of IL-6 secretion.

[0030] Figure 3 This is a graph showing the results of TNF-α secretion;

[0031] Figure 4 A graph showing the expression level of Nrf2 protein in RAW cells;

[0032] Figure 5 This is a graph showing the expression level of HO-1 protein in RAW cells. Detailed Implementation

[0033] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed in this application. Those skilled in the art can make various changes and modifications to the invention based on the disclosed content, and such changes should also fall within the scope of protection claimed in this application.

[0034] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all chemical reagents used in the embodiments of the present invention were obtained through conventional commercial means. All contents mentioned below refer to mass content.

[0035] In the following examples, arctiin was purchased from Shanghai Yuanye Biotechnology Co., Ltd., catalog number B20917; and farnesin was purchased from Shanghai Yuanye Biotechnology Co., Ltd., catalog number B20627.

[0036] The structural formula of arctiin is:

[0037]

[0038] The structural formula of acacia element is:

[0039]

[0040] Preparation of the composition of arctiin and farnesin: Arctiin and farnesin are mixed evenly according to the prescription ratio to obtain the product.

[0041] Examples 1-5

[0042] Example number Arctiin: Farnesin (mass ratio) 1 100％:0％ 2 80％:20％ 3 50％:50％ 4 20％:80％ 5 0:100％

[0043] Comparative Example 1

[0044] A mixture was prepared by combining 10 parts of burdock seed, 10 parts of belamcanda chinensis, 10 parts of platycodon grandiflorus, 10 parts of white peony root, 15 parts of perilla leaf, 10 parts of honeysuckle, 8 parts of charred hawthorn, 10 parts of astragalus root, 10 parts of polygonum cuspidatum, 8 parts of licorice root, 15 parts of fritillaria thunbergii, 5 parts of chrysanthemum, and 60 parts of black tea, as follows: The medicinal materials were weighed and mixed, pulverized at 4800 rpm for 60 minutes, and sieved to the coarsest powder, but containing no more than 40% coarse powder that could pass through a No. 4 sieve. After sieving, the mixture was mixed in a trough mixer for 30 minutes for final mixing, and then sterilized by irradiation at 60°C. The powdered medicinal materials were weighed, and ten times the amount of 60% ethanol was added (weighed and recorded); the mixture was refluxed for 1 hour, cooled, and weighed to make up for the weight loss. The mixture was centrifuged at 4000 rpm for 10 minutes, and the supernatant was concentrated to near dryness under reduced pressure. The ethanol was then evaporated to obtain the aforementioned traditional Chinese medicine composition.

[0045] Test Example Effect Measurement

[0046] 1. LPS is a classic model of macrophage inflammatory response. RAW 264.7 cells were cultured in Dulbeco modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin. Cells were incubated at 37°C and 5% CO2. RAW 264.7 cells were then sputtered at a rate of 2 × 10⁻⁶ cells / year. 4Cells were cultured at a density equal to the number of wells in 96-well plates for 24 hours. Cells were then co-incubated for 24 hours with 20 μg / mL LPS solution and samples from Examples 1, 2, 3, 4, 5, and Comparative Example 1. All experimental groups were treated with an 100 μg / mL drug concentration. The concentrations of IL-1β, IL-6, and TNF-α in the cell supernatant were detected using an ELISA kit.

[0047] like Figures 1-3 As shown, compared with unstimulated cells, LPS induction significantly increased the secretion of IL-1β, IL-6, and TNF-α in the cell supernatant. Furthermore, administration of Examples 1, 2, and 3, and Comparative Example 1, significantly inhibited the release of inflammatory factors in the cell supernatant, with Example 3 exhibiting the strongest inhibitory effect. These results indicate that, at the same dosage, significantly higher effects were achieved than when either component was used alone, or even the sum of their effects; therefore, arctiin and farnesin possess a synergistic anti-inflammatory effect.

[0048] 2. The Nrf2 / HO-1 signaling pathway is an important cellular defense mechanism that helps combat oxidative stress, inflammation, and other harmful factors. Regulation of this pathway's activity has potential significance for the treatment and prevention of various diseases. RAW 264.7 cells were cultured at 3 × 10⁻⁶ cells per day... 5 Cells were cultured in 6-well plates at a density equal to 20 μg / mL LPS solution for 24 hours. Cells were then co-incubated for 24 hours with samples from Examples 1, 2, 3, 4, 5, and Comparative Example 1, all at a drug concentration of 100 μg / mL. Cell supernatant was discarded, and proteins were extracted and separated by electrophoresis. The separated proteins were transferred to a membrane. Non-specific binding sites were blocked using skim milk powder. Nrf2 and HO-1 antibodies were added to bind to the target proteins. After washing, secondary antibodies were added for incubation to enhance the signal and visualize the target proteins. The results were recorded and analyzed using imaging equipment to detect the protein expression levels of Nrf2 and HO-1 in the cells.

[0049] like Figure 4 , Figure 5 As shown, each administration significantly inhibited the LPS-induced increase in Nrf2 and HO-1 protein expression in RAW cells, demonstrating that arctiin can regulate the Nrf2 / HO-1 signaling pathway by inhibiting Nrf2 and HO-1 expression. Comparative Example 1 also showed that the synergistic application of farnesin and arctiin can significantly enhance the activity of arctiin, with Example 3 exhibiting the strongest inhibitory activity, demonstrating that arctiin and farnesin have the strongest activity when administered at the same dose.

[0050] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. The application of a composition of arctiin and farnesin in the preparation of an anti-inflammatory drug, characterized in that, The composition of arctiin and acaciain consists of arctiin and acaciain in a mass ratio of 1:

1.

2. The application according to claim 1, characterized in that, The anti-inflammatory drug works by inhibiting the expression of inflammatory factors.

3. The application according to claim 2, characterized in that, The inflammatory factors mentioned are interleukins and tumor necrosis factor-α.

4. The application according to claim 3, characterized in that, The inflammatory factors are one or more of interleukin IL-1β, IL-6 and TNF-α.

5. The application according to claim 3, characterized in that, The interleukin mentioned is IL-6.

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