Anti-ulcerative colitis pharmaceutical composition containing anhydroicaritin

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a pharmaceutical composition for preventing and treating ulcerative colitis as well as a preparation method and application thereof. The pharmaceutical composition disclosed by the invention comprises anhydroicaritin or a pharmaceutically acceptable salt of the anhydroicaritin and N-(3-chlorphenyl)-1-(3-hydroxybenzoyl) piperidine-4-formamide or a pharmaceutically acceptable salt of the N-(3-chlorphenyl)-1-(3-hydroxybenzoyl) piperidine-4-formamide, wherein the weight ratio of the anhydroicaritin to the N-(3-chlorphenyl)-1-(3-hydroxybenzoyl) piperidine-4-formamide is (0.1-3.0): 1. The anhydroicaritin or the pharmaceutically acceptable salt of the anhydroicaritin and the N-(3-chlorphenyl)-1-(3-hydroxybenzoyl) piperidine-4-formamide or the pharmaceutically acceptable salt of the N-(3-chlorphenyl)-1-(3-hydroxybenzoyl) piperidine-4-formamide in the pharmaceutical composition disclosed by the invention have a remarkable synergistic interaction effect in the aspect of preventing or treating ulcerative colitis. The pharmaceutical composition disclosed by the invention has a remarkable treatment effect on ulcerative colitis and has important social benefits and economic values.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a drug composition for treating ulcerative colitis containing icariin. Background Technology

[0002] Ulcerative colitis (UC) is a chronic, idiopathic inflammatory bowel disease of unknown etiology. Clinically, it is characterized by chronic abdominal pain, diarrhea with mucus and bloody stools, and pathologically, persistent inflammation of the colonic mucosa and submucosa. Anti-inflammatory drugs, corticosteroids, immunosuppressants, and immunomodulators are commonly used treatments (e.g., mesalazine); however, most of these drugs have serious toxic side effects and are difficult to cure. Therefore, basic and applied research on novel drugs for the treatment of ulcerative colitis is receiving increasing attention.

[0003] Western medicine is used clinically to treat ulcerative colitis, but its efficacy is not significant, and it has significant side effects and high dependence. my country has a wide variety of traditional Chinese medicines, and its methods for treating ulcerative colitis are numerous and have a long history. Traditional Chinese medicine has unique advantages in treating diseases, such as low side effects, no dependence, no withdrawal reactions, and low cost, making it an indispensable drug for treating ulcerative colitis. Monomers of traditional Chinese medicine are its effective active ingredients, and the effects of traditional Chinese medicine are closely related to these monomers. Extensive research has been conducted on monomers such as alkaloids, flavonoids, saponins, and polysaccharides, which have played a significant role in the clinical treatment of ulcerative colitis.

[0004] Epimedium aglycone is a flavonol compound that is present in small amounts in Epimedium medicinal material. Its chemical structural formula is as follows:

[0005]

[0006] The applicant's prior patent (CN112438974A The use of icariin in the preparation of drugs for the prevention or treatment of ulcerative colitis) discloses that icariin has activity in treating ulcerative colitis.

[0007] A compound with the chemical name "N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide" has been disclosed in patent CN111518020A. This compound has MAGL inhibitor activity.

[0008] Combination therapy refers to the simultaneous or sequential use of two or more drugs to achieve a therapeutic goal. The primary aim is to enhance the efficacy of the drugs or reduce their toxic side effects; however, it can sometimes produce the opposite results. Drug combinations may exhibit synergistic, additive, or antagonistic effects.

[0009] Synergistic effect: When two or more drugs are used together, their effects are mutually enhanced, and adverse reactions are reduced or eliminated. Additive effect: The combined effect of two drugs is equal to the sum of their effects when used alone. Antagonistic effect: When two or more drugs are used together, their effects are weakened or cancel each other out. Incompatibility: When two or more drugs are used together, physical or chemical changes may occur, affecting efficacy, or even rendering the drugs ineffective or producing toxic side effects. Incompatibility can be categorized as therapeutic, physical, or chemical.

[0010] Currently, there are no reports of pharmaceutical compositions containing icariin and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide in the prevention or treatment of ulcerative colitis. Summary of the Invention

[0011] This invention discloses a pharmaceutical 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, as well as its preparation method and uses. The pharmaceutical composition of this invention, comprising icariin or a pharmaceutically acceptable salt thereof and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide or a pharmaceutically acceptable salt thereof, shows significant therapeutic efficacy for ulcerative colitis. The two components exhibit a significant synergistic effect, enhancing medication safety.

[0012] To achieve the above-mentioned objectives, the present invention adopts the following technical solution:

[0013] A combination pharmaceutical composition for treating ulcerative colitis, the combination pharmaceutical 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.

[0014] Furthermore, the mass ratio of icariin to N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide is 0.1-3.0:1.

[0015] Furthermore, the mass ratio of icariin to N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide is 0.2-2.4:1.

[0016] Furthermore, the mass ratio of icariin to N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide A is 0.2:1.

[0017] Furthermore, the mass ratio of icariin to N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide is 1.2:1.

[0018] Furthermore, the mass ratio of icariin to N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide is 2.4:1.

[0019] Furthermore, the combined pharmaceutical composition contains a pharmaceutically acceptable carrier, excipient, or other combination drug.

[0020] Furthermore, the carrier is at least one of microcapsules, microspheres, nanoparticles, and liposomes.

[0021] Furthermore, the excipients are at least one of the following: cosolvent, solubilizer, preservative, wetting agent, emulsifier, surfactant, slow-release agent, excipient, disintegrant, and lubricant.

[0022] The above-mentioned pharmaceutical composition is used in the preparation of a drug for treating ulcerative colitis.

[0023] Furthermore, the ulcerative colitis mentioned above is acute ulcerative colitis.

[0024] The combined pharmaceutical composition is administered orally or by other clinically acceptable methods.

[0025] Furthermore, the combined drug composition can be prepared into various dosage forms using conventional methods, such as oral dosage forms like powders, tablets, granules, capsules, pills, or other formulations corresponding to the route of administration.

[0026] The icariin or N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide described in this invention comprises a pharmaceutically acceptable salt thereof.

[0027] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that retains the biological potency of the free acid and free base of the specified compound and has no adverse effects in biological or other respects. The compounds of this application also include pharmaceutically acceptable salts. A pharmaceutically acceptable salt is defined as a salt formed by converting a base group in a parent compound into its salt form. Pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts containing base groups such as amine (amino) groups. Pharmaceutically acceptable salts of this application can be synthesized from a parent compound by reacting a basic group in the parent compound with 1-4 equivalents of an acid in a solvent system. Suitable salts are listed in one or more of Remingtong's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977).

[0028] As used herein, the term "treatment" and other similar synonyms include relieving, reducing, or improving symptoms of a disease or condition; inhibiting a disease or condition, such as preventing its progression; alleviating a disease or condition; improving a disease or condition; relieving symptoms caused by a disease or condition; or stopping symptoms of a disease or condition; preventing other symptoms; improving or preventing the underlying metabolic causes of symptoms; and, moreover, the term includes preventative purposes. The term also includes achieving therapeutic and / or preventative effects. A therapeutic effect refers to the cure or improvement of the underlying disease being treated. Furthermore, the cure or improvement of one or more physiological symptoms associated with the underlying disease is also a therapeutic effect; for example, an improvement is observed in a patient even though they may still be affected by the underlying disease. In terms of preventative effects, the composition may be administered to patients at risk of developing a specific disease, or to patients exhibiting one or more physiological symptoms of a disease, even if no disease diagnosis has been made.

[0029] As used herein, the terms "effective amount," "therapeutic effective amount," or "pharmaceutical effective amount" refer to the amount of at least one active substance (such as the compounds of this application) that, when taken orally, is sufficient to alleviate to some extent one or more symptoms of the disease or condition being treated. The result may be a reduction and / or relief of signs, symptoms, or causes, or any other desired change in the biological system. For example, an "effective amount" for treatment is the amount of a composition comprising the compounds disclosed herein that is clinically necessary to provide significant symptom relief. Effective amounts suitable for any individual case can be determined using techniques such as dose escalation testing.

[0030] This invention combines icariin or a pharmaceutically acceptable salt thereof with N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide or a pharmaceutically acceptable salt thereof to relieve and treat ulcerative colitis. The combination of the two components has a significant synergistic effect, significantly improves the symptoms of loose stools and rectal bleeding induced by DSS, improves the weight loss caused by DSS, and improves colonic shortening.

[0031] Compared with the prior art, the present invention has the following advantages and beneficial effects.

[0032] The active ingredient of the pharmaceutical composition provided by this invention comprises icariin 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 this invention, the combined use of icariin or a pharmaceutically acceptable salt thereof and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide or a pharmaceutically acceptable salt thereof at their respective effective dosage ratios is more effective than using either ingredient alone, exhibiting a synergistic effect.

[0033] The pharmaceutical composition of the present invention, containing icariin or a pharmaceutically acceptable salt thereof and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide or a pharmaceutically acceptable salt thereof, can achieve the therapeutic effect of ulcerative colitis while reducing drug dosage, reducing drug toxicity and side effects, and has high safety.

[0034] The pharmaceutical composition of the present invention has the potential to be developed into a drug for the prevention and treatment of ulcerative colitis, providing more treatment options for the prevention or treatment of ulcerative colitis in clinical practice, and has significant social and economic benefits. Detailed Implementation

[0035] The technical solution of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0036] Epimedium flavonoids exist mainly in the form of glycosides in the plant Epimedium, while icariin is present in very low amounts in the original plant and is difficult to directly isolate and purify. Therefore, icariin can be prepared by hydrolyzing the glycosidic bonds in the flavonoid glycosides. Methods for preparing icariin from icariin include enzymatic hydrolysis, acid hydrolysis, and a combination of enzymatic and acid hydrolysis. Reported methods for hydrolyzing glycosidic bonds include: (1) chemical methods, such as acid and alkaline hydrolysis. Acid hydrolysis often uses hydrochloric acid, sulfuric acid, or nitric acid, while alkaline hydrolysis often uses sodium hydroxide or potassium hydroxide. This method often uses the hydrolysis of the 3-rhamnoside and 7-glucose glycosidic bonds of icariin to obtain icariin. These studies use icariin as the reaction substrate. (2) Biotransformation method, such as using Epimedium as an inducer for enzyme production in the fermentation of bacteria, molds, and yeasts to prepare a mold-containing liquid, and then using this liquid to convert Epimedium flavonoid glycosides into low-glycosides or aglycones. Epimedium flavonoid glycosides are diverse, and their glycosidic bond structures are very complex, mainly composed of monosaccharide glycosidic bonds such as α-L-rhamnosyl glycosidic bonds, β-D-glucose glycosidic bonds, and β-D-xylose glycosidic bonds, as well as disaccharide glycosidic bonds composed of them. Chinese Patent CN201711485889.7, entitled "A Method for Preparing Epimedium Aglycones," provides a method for preparing epimedium aglycones. Based on the above methods, epimedium aglycones can be prepared.

[0037] The icariin in this invention can be prepared from icariin, or it can be prepared by chemical synthesis or purchased commercially.

[0038] The drugs selected for the pharmacodynamic tests described below are those obtained from the representative formulations and preparation methods of this invention. The inventors also conducted pharmacodynamic tests on drugs obtained from other formulations and preparation methods included in this invention. The experimental results show that drugs obtained from other formulations and preparation methods have the same or similar effects, but due to space limitations, they are not listed here. In addition, the pharmacodynamic tests described below only use some representative animal models as examples to verify the efficacy of this invention.

[0039] Pharmacological Example 1: Effect of the pharmaceutical composition of the present invention containing N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide and icariin on a mouse model of ulcerative colitis.

[0040] 1. Laboratory animals

[0041] Eighty male C57BL / 6J mice, weighing 20-22g, were provided by Beijing Huafukang Biotechnology Co., Ltd., Animal Qualification Certificate No.: AN-IACUC-2023-094. The mice were housed in a strictly controlled SPF-grade animal enclosure with a temperature of 20-26℃, humidity of 40-70%, a light-dark cycle of 12:12h, and were given growth and reproduction feed with free access to food.

[0042] 2. Main reagents

[0043] N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide (hereinafter referred to as compound A) was prepared according to the method disclosed in CN111518020A and was provided by Shandong New Era Pharmaceutical Co., Ltd.

[0044] Epimedium glycoside was provided by Shandong New Era Pharmaceutical Co., Ltd.

[0045] 3. Experimental methods and experimental grouping:

[0046] Eighty male C57BL / 6J mice, aged 20-22g and 6-8 weeks, were randomly divided into 10 groups: normal group, model group, low-dose icariin group, medium-dose icariin group, high-dose icariin group, low-dose compound A group, high-dose compound A group, composition A group, composition B group, and composition C group, with 8 mice in each group.

[0047] The normal group had free access to normal water, while the other groups had free access to 1.5% DSS solution for modeling (1.5g DSS dissolved in 100mL distilled water). After 7 days of modeling, the modeling groups had free access to normal water for 7 days. This 14-day period was one modeling cycle, and four consecutive cycles were conducted. The drugs were administered simultaneously with the modeling process, and the experimental drugs were administered for four cycles. Dissection was performed on the fourth day of the recovery period in the fourth cycle.

[0048] The administration of drugs to each group is as follows:

[0049] Low-dose icariin group: Icariin 6 mg / (kg·d) was administered by gavage;

[0050] Medium-dose group of icariin: 12 mg / (kg·d) of icariin was administered by gavage;

[0051] High-dose icariin group: Icariin 24 mg / (kg·d) was administered by gavage;

[0052] Low-dose group of compound A: N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide 10 mg / (kg·d) was administered by gavage;

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

[0054] Composition A: 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: 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: Icariin 24 mg / (kg·d) and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide 10 mg / (kg·d) were administered by gavage.

[0057] Mice in the normal group and the model group were administered the same dose of solvent (0.5% sodium carboxymethyl cellulose) by gavage.

[0058] The experimental data were analyzed using SPSS 19.0 statistical software. Quantitative data were expressed as mean ± standard deviation. The results indicate that the difference was statistically significant using t-tests and ANOVA, with P < 0.05.

[0059] 4. General observation

[0060] After administration, the mice were weighed, and their diarrhea, feces, and bloody stools were observed. The mice were scored 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 P4508B1 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 fecal index and bloody stool index were recorded.

[0061] Stool consistency index: 0 for normally formed pellet stool, 1 for soft but still formed stool, 2 for soft but still formed stool, 3 for very soft and moist stool, and 4 for watery diarrhea.

[0062] Rectal bleeding index: 0 points for negative rectal bleeding, 1 point for weakly positive rectal bleeding, 2 points for positive rectal bleeding, 3 points for visible bloodstains in stool, and 4 points for massive bleeding.

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

[0064] Fifty-six days after treatment, the mouse colon was isolated and its length was recorded by photograph.

[0065] 6. Experimental Results

[0066] 6.1 Bloody stool and loose stool in mice

[0067] Table 1 shows that the fecal hemorrhage index was significantly increased in the ulcerative colitis model group compared with the normal group. Compared with the model group, the various doses of icariin, compound A, and the combined composition significantly improved fecal hemorrhage in mice. The combined composition group showed better improvement in fecal hemorrhage, significantly superior to the various doses of icariin and compound A. This indicates that icariin and compound A have a significant synergistic effect in improving fecal hemorrhage in mice with ulcerative colitis.

[0068] Compared with the normal group, the loose stool index of mice in the ulcerative colitis model group was significantly increased. Compared with the model group, the various doses of icariin, compound A, and the combined composition significantly improved the loose stool condition in mice. The combined composition group showed better improvement in loose stool, significantly superior to the individual doses of icariin and compound A. This indicates that icariin and compound A have a significant synergistic effect in improving loose stool in mice with ulcerative colitis.

[0069] Table 1 Comparison of fecal blood index and loose stool index among different groups of mice ( n=8)

[0070]

[0071]

[0072] Note: Compared with the normal group ### P<0.001;

[0073] Compared with the model group, * P<0.05, ** P<0.01, *** P<0.001.

[0074] 6.2 Mouse body weight and colon length

[0075] Table 2 shows that the body weight of mice in the ulcerative colitis model group was significantly lower than that of the normal group. Compared with the model group, the various doses of icariin, compound A, and the combined composition significantly increased the body weight of mice. The combined composition group showed a better effect in increasing the body weight of mice, significantly better than the individual doses of icariin and compound A. This indicates that icariin and compound A have a significant synergistic effect in increasing the body weight of mice with ulcerative colitis.

[0076] Compared with the normal group, the colon length of mice in the ulcerative colitis model group was significantly shortened. Compared with the model group, the various doses of icariin, compound A, and the combined composition significantly increased the colon length of mice. The combined composition group showed a better effect in increasing the colon length of mice, significantly superior to the various doses of icariin and compound A. This indicates that icariin and compound A have a significant synergistic effect in increasing the colon length of mice with ulcerative colitis.

[0077] Table 2 Comparison of body weight ratio and colon length of mice in each group ( n=8)

[0078]

[0079] Note: Compared with the normal group ### P<0.001;

[0080] Compared with the model group, * P<0.05, ** P<0.01, *** P<0.001.

[0081] In summary, the combined use of icariin and N-(3-chlorophenyl)-1-(3-hydroxybenzoyl)piperidine-4-carboxamide significantly improved hematochezia and loose stools in mice, increased mouse body weight, and increased colon length in mice, demonstrating a significant synergistic effect in the treatment of ulcerative colitis.

[0082] The above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description, and any obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

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 pharmaceutical 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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