Anti-ulcerative colitis pharmaceutical composition containing icaritin

Abstract

Disclosed are a pharmaceutical composition for preventing and treating ulcerative colitis, a preparation method therefor, and use thereof. The pharmaceutical composition comprises icaritin or a pharmaceutically acceptable salt thereof and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide or a pharmaceutically acceptable salt thereof, and the weight ratio of icaritin to N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide is 0.1-3.0:1. Icaritin or the pharmaceutically acceptable salt thereof and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide or the pharmaceutically acceptable salt thereof in the pharmaceutical composition exhibit a significant synergistic effect in preventing or treating ulcerative colitis, resulting in a notable therapeutic effect on ulcerative colitis.

Description

Anti-ulcerative colitis medicine composition containing icariogenin TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and particularly relates to an anti-ulcerative colitis medicine composition containing icariogenin. BACKGROUND

[0002] Ulcerative colitis (UC) is a chronic, idiopathic intestinal inflammatory disease, the cause of which is unknown. Clinically, the main characteristics are chronic abdominal pain, diarrhea with mucus and blood stool. Pathologically, the main characteristic is persistent inflammation of the mucosal layer and submucosal layer of the colon. Anti-inflammatory drugs, corticosteroids, immunosuppressants and immunomodulators are currently commonly used therapeutic drugs (such as mesalazine), however, most of these drugs have serious toxic side effects and are difficult to cure. Therefore, basic and applied research on new drugs for treating ulcerative colitis is attracting attention.

[0003] Clinically, western medicine is used to treat ulcerative colitis, but the curative effect is not obvious, the side effects are large, and the dependence is strong. There are many types of traditional Chinese medicines in China, and there are many methods for treating ulcerative colitis with traditional Chinese medicines, which have a long history. Traditional Chinese medicine has unique advantages in treating diseases, such as low side effects, no dependence, no withdrawal reaction, and low price, and is an indispensable drug for treating ulcerative colitis. Traditional Chinese medicine monomers are effective active ingredients of traditional Chinese medicine, and the action of traditional Chinese medicine is closely related to traditional Chinese medicine monomers. Among them, the research on alkaloids, flavonoids, saponins, polysaccharides and other monomers is extensive, and has played a great role in the clinical treatment of ulcerative colitis.

[0004] Icariogenin belongs to flavonol compounds, and exists in a small amount in icariin medicinal materials, and its chemical structural formula is as follows:

[0005] The prior patent (CN112438974A Application of icariogenin in preparation of medicine for preventing or treating ulcerative colitis) of the applicant discloses that icariogenin has activity in treating ulcerative colitis.

[0006] The compound with the chemical name of "N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide" has been disclosed in the patent CN111518020A, and the compound has MAGL inhibitor activity.

[0007] Combination drug refers to the simultaneous or sequential use of two or more drugs for the purpose of treatment, and the result is mainly to increase the curative effect of the drug or to reduce the toxic side effects of the drug, but sometimes it may also have the opposite result. Drug combination may show synergistic effect, additive effect or antagonistic effect.

[0008] Synergism: the combined effect of two or more drugs is greater than the sum of their individual effects. Additive effect: the combined effect of two drugs is equal to the sum of their individual effects. Antagonism: the combined effect of two or more drugs is less than the sum of their individual effects. Incompatibility: two or more drugs combined will have physical or chemical changes, affecting the efficacy, even invalid or produce side effects. Can be divided into curative effect, physical and chemical incompatibility.

[0009] Currently, there is no report on the use of the pharmaceutical composition containing icariin and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide in the prevention or treatment of ulcerative colitis. SUMMARY

[0010] The present application discloses a pharmaceutical composition containing icariin or a pharmaceutically acceptable salt thereof and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide or a pharmaceutically acceptable salt thereof, and its preparation method and use. The pharmaceutical composition containing icariin or a pharmaceutically acceptable salt thereof and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide or a pharmaceutically acceptable salt thereof of the present application has a significant therapeutic effect on ulcerative colitis, and the two components have a significant synergistic effect, enhancing the safety of the drug.

[0011] In order to achieve the above-mentioned application purposes, the present application adopts the following technical solutions:

[0012] A combined drug composition for treating ulcerative colitis, the combined drug composition comprising icariin or a pharmaceutically acceptable salt thereof and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide or a pharmaceutically acceptable salt thereof.

[0013] Further, the mass ratio of icariin and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide is 0.1-3.0:1.

[0014] Further, the mass ratio of icariin and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide is 0.2-2.4:1.

[0015] Further, the mass ratio of icariin and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide A is 0.2:1.

[0016] Further, the mass ratio of icariin and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide is 1.2:1.

[0017] Further, the mass ratio of the icariin and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide is 2.4:1.

[0018] Further, the combination drug composition contains a pharmaceutically acceptable carrier, adjuvant or other combination drug.

[0019] Further, the carrier is at least one of microcapsule, microsphere, nanoparticle, liposome.

[0020] Further, the adjuvant is at least one of solubilizing agent, solubilizer, preservative, wetting agent, emulsifier, surfactant, sustained-release agent, excipient, disintegrant, lubricant.

[0021] The above drug composition is used for preparing a drug for treating ulcerative colitis.

[0022] Further, the ulcerative colitis is acute ulcerative colitis.

[0023] The administration mode of the combination drug composition is oral or other clinically acceptable mode.

[0024] Further, the combination drug composition can be prepared into various dosage forms by conventional methods, such as powder, tablet, granule, capsule, pill and other oral dosage forms, or other preparations corresponding to the administration route.

[0025] The icariin or N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide, including pharmaceutically acceptable salts thereof, of the present application.

[0026] The term "pharmaceutically acceptable salt" as used herein refers to salts that retain the biological effectiveness and none of the adverse effects of the free acids and free bases of the designated compounds. The compounds of the present application also include pharmaceutically acceptable salts. Pharmaceutically acceptable salts refer to salts of the base moieties converted from the base moieties in the parent compounds. Pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic groups such as amine (ammonia) groups. The pharmaceutically acceptable salts of the present application can be synthesized from the parent compound by reacting the basic moiety with 1-4 equivalents of the acid in a solvent system. Suitable salts are listed in one or more of Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977).

[0027] The terms "treatment" and other similar synonymous terms as used herein include alleviating, abating, or ameliorating a disease or condition, inhibiting the disease or condition, e.g., arresting the development of the disease or condition, relieving the disease or condition, causing the disease or condition to regress, relieving the symptoms resulting from the disease or condition, or stopping the symptoms of the disease or condition, preventing other symptoms, ameliorating or preventing the underlying metabolic cause of symptoms, and in addition the term encompasses the purpose of preventing. The term also includes obtaining a therapeutic benefit and / or a prophylactic benefit. By therapeutic benefit is meant healing or improvement of the underlying disease. In addition, healing or improvement of one or more of the physiological symptoms associated with the underlying disease is a therapeutic benefit, e.g., an improvement of a patient's condition, even though the patient can still be affected by the disease. In terms of a prophylactic benefit, the compositions can be administered to a patient at risk of developing a particular disease, or to a patient reporting one or more of the physiological symptoms of a disease, even though the patient can not yet be diagnosed with the disease.

[0028] The terms "effective amount", "therapeutically effective amount", or "pharmaceutically effective amount" as used herein refer to the amount of at least one active agent (e.g., a compound of the application) that, when administered, is sufficient to relieve to some extent one or more of the symptoms of the disease or condition being treated. The result can be reduction and / or alleviation of the signs, symptoms, or causes of a disease or disorder, or any other desired alteration of a biological

[0029] The present application can alleviate and treat ulcerative colitis by using the combination of icariin or its pharmaceutically acceptable salt and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide or its pharmaceutically acceptable salt. The combination of the two components has a significant synergistic effect, significantly improves the symptoms of loose stool and blood in stool caused by DSS, improves the weight loss caused by DSS, and improves the shortening of the colon.

[0030] Compared with the prior art, the present application has the following advantages and beneficial effects.

[0031] The active ingredient of the pharmaceutical composition provided by the present application consists of icariogenin or a pharmaceutically acceptable salt thereof and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide or a pharmaceutically acceptable salt thereof, which can effectively relieve and treat ulcerative colitis. In the pharmaceutical composition of the present application, icariogenin or a pharmaceutically acceptable salt thereof and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide or a pharmaceutically acceptable salt thereof are combined in a proportion of an effective dose of each, and the effect is better than that of using an effective dose of each alone, and has a synergistic effect.

[0032] The pharmaceutical composition containing icariogenin or a pharmaceutically acceptable salt thereof and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide or a pharmaceutically acceptable salt thereof of the present application can reduce the amount of drugs, reduce the toxic side effects of drugs, and has high safety while achieving the therapeutic effect of ulcerative colitis.

[0033] The pharmaceutical composition of the present application has the prospect of developing into a drug for preventing and treating ulcerative colitis, and provides more treatment options for preventing or treating ulcerative colitis in the clinic, and has important social and economic values. DETAILED DESCRIPTION

[0034] The technical solutions of the present application will be described below clearly and completely. Obviously, the described embodiments are part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor are within the scope of protection of the present application.

[0035] The flavonoid components in Epimedium are mainly in the form of glycosides, and the icariin aglycone is difficult to be directly isolated and purified from the original plant due to its low content. Therefore, the icariin aglycone can be prepared by hydrolyzing the glycosidic bond of the flavonoid glycosides in Epimedium. The methods for preparing icariin aglycone from icariin include enzymatic hydrolysis, acid hydrolysis, and the combination of enzymatic hydrolysis and acid hydrolysis. The reported methods for hydrolyzing the glycosidic bond include: (1) chemical methods, such as acid hydrolysis and alkaline hydrolysis. The acid hydrolysis method mainly uses hydrochloric acid, sulfuric acid or nitric acid, and the alkaline hydrolysis method mainly uses sodium hydroxide or potassium hydroxide. This method mainly hydrolyzes the 3-rhamnose glycosidic bond and the 7-glucose glycosidic bond of icariin to obtain icariin aglycone. These studies use icariin as the reaction substrate. (2) biological transformation method, such as using Epimedium as the fermentation enzyme inducer of bacteria, molds and yeasts to prepare mold liquid, and then using the mold liquid to convert flavonoid glycosides in Epimedium into low glycosides or aglycone. There are many types of flavonoid glycosides in Epimedium, and the structure of the glycosidic bond is very complex, which mainly includes α-L-rhamnose glycosidic bond, β-D-glucose glycosidic bond, β-D-xylose glycosidic bond and disaccharide glycosidic bond composed of them. Chinese patent CN201711485889.7, entitled “Preparation method of icariin aglycone” provides a preparation method of icariin aglycone. Based on the above-mentioned preparation methods, icariin aglycone can be prepared.

[0036] The icariin aglycone in the present application can be prepared from icariin, or can be prepared by chemical synthesis method, or can be purchased on the market.

[0037] The drugs selected for the pharmacodynamic test described below are the drugs obtained by the representative formula and preparation method of the present application. The inventors have also conducted pharmacodynamic tests on other formulas and drugs obtained by the preparation methods included in the present application, and the experimental results show that the drugs obtained by other formulas and preparation methods have the same or similar effects. However, due to the limitation of the length of the article, they are not listed one by one. In addition, the pharmacodynamic test described below only takes some representative animal models as examples to verify the efficacy of the present application.

[0038] Effect of the pharmaceutical composition containing N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide and icariin aglycone of the present application on the mouse model of ulcerative colitis

[0039] 1. Experimental animals

[0040] Male C57BL / 6J mice 80, body weight 20-22 g, provided by Beijing Huafukang Biotechnology Co., Ltd., animal qualification certificate number: AN-IACUC-2023-094. The mice were raised in SPF level animal feeding room with strictly controlled environmental conditions, temperature 20-26℃, humidity 40-70%, light and dark cycle 12:12h, growth and reproduction feed, free drinking.

[0041] 2. Main reagents

[0042] N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide (hereinafter referred to as compound A) was prepared according to the method disclosed in CN111518020A, provided by Shandong New Age Pharmaceutical Co., Ltd.

[0043] The structural formula of compound A is as follows:

[0044] Icariin, provided by Shandong New Age Pharmaceutical Co., Ltd.

[0045] 3. Experimental methods and experimental grouping:

[0046] 80 C57BL / 6J 20-22g / 6-8 week old male mice were randomly divided into 10 groups, namely normal group, model group, icariin low dose group, icariin medium dose group, icariin high dose group, compound A low dose group, compound A high dose group, combination A group, combination B group, and combination C group, 8 mice in each group.

[0047] The normal group freely drank normal water, and the rest of the groups freely drank 1.5% DSS solution for modeling (1.5g DSS was dissolved in 100mL distilled water). After 7 days of modeling, the modeling groups freely drank normal water for 7 days. This 14-day period was a modeling cycle, and four cycles were performed in succession. Modeling was performed simultaneously with drug administration, and the experiment was performed for four cycles. Autopsy was performed on the fourth day of the recovery period of the fourth cycle.

[0048] The dosages of each group are as follows:

[0049] Icariin low dose group: intragastrically administered with icariin 6mg / (kg.d);

[0050] Icariin medium dose group: intragastrically administered with icariin 12mg / (kg.d);

[0051] Icariin high dose group: intragastrically administered with icariin 24mg / (kg.d);

[0052] Compound A low dose group: intragastrically administered with N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide 10mg / (kg.d);

[0053] Compound A high dose group: N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide 30 mg / (kg.d) was administered by gavage;

[0054] Composition A group: Icariin 6 mg / (kg.d) and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide 30 mg / (kg.d) were administered by gavage;

[0055] Composition B group: Icariin 12 mg / (kg.d) and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide 10 mg / (kg.d) were administered by gavage;

[0056] Composition C group: Icariin 24 mg / (kg.d) and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide 10 mg / (kg.d) were administered by gavage.

[0057] The normal group and model group mice were administered by gavage with the same dose of solvent (0.5% carboxymethylcellulose sodium).

[0058] The experimental data were analyzed using SPSS 19.0 statistical software, and the measurement data were expressed as mean ± standard deviation The t test and variance analysis were used, and P<0.05 was considered statistically significant.

[0059] 4. General observation

[0060] After starting the administration, the body weight of the mice was weighed, and the diarrhea, feces, and bloody stool of the mice were observed, and the score was made according to the literature method (Chen L, Jiao T, Liu W, Luo Y, Wang J, Guo X, Tong X, Lin Z, Sun C, Wang K, He Y, Zhang Y, Xu H, Wang J, Zuo J, Ding Q, He S, Gonzalez FJ, Xie C. Hepatic cytochrome P450 8B1 and cholic acid potentiate intestinal epithelial injury in colitis by suppressing intestinal stem cell renewal. Cell Stem Cell. 2022 Sep 1; 29(9): 1366-1381.e9. doi: 10.1016 / j.stem.2022.08.008), and the feces index and bloody stool index were recorded.

[0061] Loose stool index: normal formed granular stool consistency score 0, soft but still formed stool 1, soft but still formed stool 2, very soft and wet stool 3, watery diarrhea 4.

[0062] Blood in stool index: blood in stool negative 0 points, weakly positive blood in stool 1 point, positive blood in stool 2 points, blood visible in stool 3 points, massive hemorrhage 4 points.

[0063] 5. Detecting the length of the colon and the spleen index of mice

[0064] After 56 days of treatment, the colon of the mice was isolated and the length of the colon was recorded by taking a photo.

[0065] 6. Experimental results

[0066] 6.1 Blood in stool and loose stool of mice

[0067] As can be seen from Table 1, compared with the normal group, the blood in stool index of the ulcerative colitis model group of mice increased significantly. Compared with the model group, the icariin and compound A each dose group and the composition each group significantly improved the blood in stool of mice. The effect of improving the blood in stool of mice of the composition each group was better, significantly better than that of the icariin and compound A each dose group. It is proved that icariin and compound A have significant synergistic effect in improving the blood in stool of ulcerative colitis mice.

[0068] Compared with the normal group, the loose stool index of the ulcerative colitis model group of mice increased significantly. Compared with the model group, the icariin and compound A each dose group and the composition each group significantly improved the loose stool of mice. The effect of improving the loose stool of mice of the composition each group was better, significantly better than that of the icariin and compound A each dose group. It is proved that icariin and compound A have significant synergistic effect in improving the loose stool of ulcerative colitis mice.

[0069] Comparison of blood in stool index and loose stool index of mice in each group of Table 1 n=8)

[0070] Note: compared with the normal group, ### P<0.001;

[0071] Compared with the model group, * P<0.05, ** P<0.01, *** P<0.001.

[0072] 6.2 Body weight and colon length of mice

[0073] As shown in Table 2, compared with the normal group, the body weight of the ulcerative colitis model group mice was significantly decreased. Compared with the model group, the icariin and compound A each dose group and the composition each group significantly increased the body weight of the mice. The effect of the composition each group on increasing the body weight of the mice was better, and was significantly better than that of the icariin and compound A each dose group. It was indicated that icariin and compound A had a significant synergistic effect on increasing the body weight of the ulcerative colitis mice.

[0074] Compared with the normal group, the colon length of the ulcerative colitis model group mice was significantly shortened. Compared with the model group, the icariin and compound A each dose group and the composition each group significantly increased the colon length of the mice. The effect of the composition each group on increasing the colon length of the mice was better, and was significantly better than that of the icariin and compound A each dose group. It was indicated that icariin and compound A had a significant synergistic effect on increasing the colon length of the ulcerative colitis mice.

[0075] Comparison of the body weight ratio and colon length of the mice in each group in Table 2 n=8

[0076] Note: compared with the normal group, ### P<0.001;

[0077] Compared with the model group, * P<0.05, ** P<0.01, *** P<0.001.

[0078] In summary, icariin and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide in combination can significantly improve the blood in the stool and the loose stool of the mice, increase the body weight of the mice, increase the colon length of the mice, and have a significant synergistic effect in treating ulcerative colitis.

[0079] The above examples are only examples for clearly illustrating, but not limitation to the embodiments. For those skilled in the art, other different forms of changes or variations can be made on the basis of the above description, and the obvious changes or variations derived therefrom are still within the protection scope of the present application.

Claims

1. A combination pharmaceutical composition for treating ulcerative colitis, characterized by comprising, The combined pharmaceutical composition comprises icariin or a pharmaceutically acceptable salt thereof and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide or a pharmaceutically acceptable salt thereof.

2. The combined pharmaceutical composition for treating ulcerative colitis according to claim 1, wherein The mass ratio of icariin and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide is 0.1-3.0:

1.

3. The combined pharmaceutical composition for treating ulcerative colitis according to claim 1, wherein The mass ratio of icariin and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide is 0.2-2.4:

1.

4. The combined pharmaceutical composition for treating ulcerative colitis according to claim 1, wherein The mass ratio of icariin and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide is 0.2:1, 1.2:1 or 2.4:

1.

5. The combination of claim 1-4 for use in the treatment of ulcerative colitis, wherein The combined pharmaceutical composition contains a pharmaceutically acceptable carrier or excipient.

6. The combination medicament composition for treating ulcerative colitis according to claim 5, wherein The carrier is at least one of microcapsules, microspheres, nanoparticles, liposomes; the excipient is at least one of a cosolvent, a solubilizer, a preservative, a wetting agent, an emulsifying agent, a surfactant, a sustained-release agent, an excipient, a disintegrating agent, a lubricant.

7. The pharmaceutical composition of any one of claims 1-6 for use in the preparation of a medicament for preventing or treating ulcerative colitis.

8. Use according to claim 7, characterized in that, The ulcerative colitis is acute ulcerative colitis.

9. Use according to claim 7 or 8, characterized in that, The combined pharmaceutical composition is administered orally.

10. Use according to claim 7 or 8, characterized in that, The dosage form of the combined pharmaceutical composition is powder, tablet, granule, capsule or pill. The combined pharmaceutical composition contains a pharmaceutically acceptable carrier or excipient. The carrier is at least one of microcapsules, microspheres, nanoparticles, liposomes; the excipient is at least one of a cosolvent, a solubilizer, a preservative, a wetting agent, an emulsifying agent, a surfactant, a sustained-release agent, an excipient, a disintegrating agent, a lubricant.

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