CN104771408B - The purposes of notoginsenoside Ft1 - Google Patents

Abstract

The invention relates to the technical field of medicine, in particular to the use of notoginseng saponin Ft1. The invention discloses the use of notoginsenoside Ft1, prodrugs and pharmaceutically acceptable salts thereof or plant extracts containing notoginsenoside Ft1 in the preparation of medicines or health care products for preventing or treating ulcerative colitis. The present invention finds for the first time that notoginseng saponin Ft1 can significantly improve symptoms such as weight loss, bloody stool, colon shortening, and histopathological damage in mice with experimental ulcerative colitis, providing a new option for clinical treatment of ulcerative colitis, and has good application prospect.

Description

The use of notoginseng saponin Ft1

technical field

The invention relates to the technical field of medicine, in particular to the use of notoginseng saponin Ft1.

Background technique

Ulcerative colitis (UC) is a chronic non-specific inflammatory disease of the colon and rectum with unknown etiology. The main symptoms are diarrhea, mucus pus and blood, abdominal pain and tenesmus. Ulcer formation is a common and intractable disease of the digestive tract with pathological characteristics. In severe cases, there may be extraintestinal symptoms such as fever, arthritis, iritis, erythema nodosum, and serious complications such as colon bleeding, perforation, and cancer. The acute fulminant type of this disease has a high mortality rate, and the chronic persistent type is prone to canceration. It is reported that the incidence rate is 5-20 times higher than that of normal people, with an average of 3.5%-7%. The canceration rate of patients is 20% at 10 years, and 40% at 25 years or more, especially those with long course of disease, wide range of lesions, and younger age, which are recognized as precancerous lesions of colon cancer. The malignant transformation rate in the chronic phase of this disease is quite high, and the cancer that occurs in ulceration is highly malignant and often metastasizes in the early stage. It has been listed by the World Health Organization as one of the modern intractable diseases. At present, sulfasalazine, 5-aminosalicylic acid (5-ASA) sustained-release dosage forms, and corticosteroids are still the main drugs for clinical treatment of ulcers, and patients often need long-term medication for maintenance, often recurrent attacks.

Notoginseng saponin Ft1 (Vitexin, Apigenin-8-C-β-D-glucopyranoside) is a natural saponin compound mainly present in notoginseng. Studies have found that notoginseng saponin Ft1 has anti-oxidation, myocardial protection, anti-tumor and other activities, and its toxicity is extremely low.

Contents of the invention

The technical problem to be solved by the present invention is to provide an active drug for treating ulcerative colitis and a pharmaceutical composition thereof.

Therefore, the present invention discloses the use of notoginsenoside Ft1, prodrugs and pharmaceutically acceptable salts thereof, or plant extracts containing notoginsenoside Ft1 in the preparation of medicines or health care products for preventing or treating ulcerative colitis.

In some embodiments, the plant extract containing notoginseng saponin Ft1 is the alcoholic extract of leaves or roots of Araliaceae plants, including but not limited to the alcoholic extract of Radix Notoginseng.

In another aspect, the present invention provides a pharmaceutical composition for treating ulcerative colitis, said composition comprising:

(a) containing 1-99% by weight of notoginseng saponin Ft1 and its pharmaceutically acceptable salts as active ingredients;

(b) parts by weight are pharmaceutically acceptable carriers as the balance.

In another preferred example, the said notoginsenoside Ft1 can form salts with inorganic acids or organic acids, such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, acetic acid, maleic acid, rich Salts of malic acid, citrate, citric acid, oxalic acid, succinic acid, tartaric acid, malic acid, mandelic acid, trifluoroacetic acid, pantothenic acid, methanesulfonic acid or p-toluenesulfonic acid.

In another preferred example, the dosage form of the pharmaceutical composition is selected from: injections, sterile powders for injections, tablets, capsules, spirits, powders, granules, syrups, solutions, tinctures, aerosols , powder spray or suppository, preferably tablet or capsule.

In another aspect, the present invention provides a treatment method for preventing or treating ulcerative colitis, which comprises administering an effective dose of the pharmaceutical composition of the present invention or the plant extract containing notoginseng saponin Ft1 to patients with ulcerative colitis.

The present invention finds for the first time that notoginseng saponin Ft1 can significantly improve symptoms such as weight loss, bloody stool, colon shortening, and histopathological damage in mice with experimental ulcerative colitis, providing a new option for clinical treatment of ulcerative colitis, and has good application prospect.

Description of drawings

The effect of Fig. 1 notoginseng saponin Ft1 on the body weight of mice with ulcerative colitis;

The effect of Fig. 2 notoginseng saponin Ft1 on the incidence of bloody stool in mice with ulcerative colitis;

Figure 3 Effect of notoginseng saponin Ft1 on the colon length of mice with ulcerative colitis, the overall appearance of the colon in Figure 3A, and the colon length in Figure 3B;

Figure 4 Effect of notoginseng saponin Ft1 on colon histopathology of mice with ulcerative colitis, HE staining of mouse colon in Figure 4A, histopathological scoring in Figure 4B;

(In the above figure: a represents the normal control group, b represents the model control group, c represents the SASP group, d represents the notoginsenoside Ft1 group)

Detailed ways

The molecular formula of the notoginsenoside Ft1 of the present invention is: C47H80O17, and the molecular weight is: 917.13.

The notoginseng saponin Ft1 of the present invention can be purchased commercially from Shanghai Chunyou Biotechnology Co., Ltd., Chengdu Master Biotechnology Co., Ltd., etc. Its purity is in line with pharmaceutical standards. The purity of the notoginseng saponin Ft1 of the present invention is ≥98%.

The active compound notoginseng saponin Ft1 of the present invention can be extracted from plants containing notoginseng saponin Ft1, such as alcoholic extracts of leaves or roots of Araliaceae plants, including but not limited to alcoholic extracts of notoginseng roots.

The term "pharmaceutically acceptable salt" refers to certain salts of the above-mentioned compounds that can maintain their original biological activity and are suitable for medical use. The pharmaceutically acceptable salts of the compounds of the present invention can be formed in two forms: one is the salt formed with acid; the other is the salt formed with alkali or alkali metal. Acids which form pharmaceutically acceptable salts with the compounds described herein include inorganic and organic acids. Suitable inorganic acids include: hydrochloric acid, sulfuric acid and phosphoric acid. Suitable organic acids may be selected from aliphatic, cycloaliphatic, aromatic, heterocyclic carboxylic and sulfonic organic acids; examples include, but are not limited to: formic acid, acetic acid, propionic acid, succinic acid, glycolic acid, glucose acid, lactic acid, malic acid, tartaric acid, glycine, arginine, citric acid, fumaric acid, alkylsulfonic acid, etc. The alkali metals that form pharmaceutically acceptable salts with the compounds of the present invention include: lithium, sodium, potassium, magnesium, calcium, aluminum or zinc and the like.

The present invention includes the compounds described herein and all possible isomeric forms thereof. Includes: diastereomers, mirror-image isomers, tautomers, geometric isomers of "E" or "Z" configuration isomers, and the like.

The present invention includes the compounds of the present invention and their possible racemates or/and mirror isomers/or/and mixtures of diastereomers.

In addition, the compounds of the present invention also cover solvated and unsolvated forms of the compounds in application. Accordingly, the formulas include compounds of the indicated configuration, including hydrated and anhydrous forms thereof.

In addition to the compounds described herein, various embodiments include pharmaceutically acceptable salts, prodrugs and active metabolites of these compounds. and pharmaceutically acceptable salts of such metabolites.

A "prodrug" is a derivative of the compound of the present invention, which is transformed into the compound of the present invention by means of metabolism in vivo (for example: by hydrolysis, reduction or oxidation). For example, the compounds of the invention can be reacted with acids to prepare the corresponding esters. The corresponding ester is a prodrug, which can be hydrolyzed into the parent drug in vivo. Acids suitable for preparing "prodrugs" include, but are not limited to: acetic acid, citric acid, lactic acid, tartaric acid, malonic acid, oxalic acid, salicylic acid, succinic acid, fumaric acid, maleic acid, methylene Base-bis-β-hydroxynaphthoic acid, gentisic acid, isethionic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, etc.

use

The compounds of the present invention or their pharmaceutically acceptable salts or prodrugs or their isomers or plant extracts containing them can be used to prevent or treat ulcerative colitis. A treatment method for preventing or treating ulcerative colitis, comprising administering an effective dose of the pharmaceutical composition of the present invention or the plant extract containing it to patients with ulcerative colitis.

"Effective dose" or "therapeutic amount" both refer to an amount sufficient to produce a therapeutic effect. An effective amount may be administered in one or more divided doses. Usually, the effective amount is enough to alleviate, ameliorate, stabilize, slow down or delay the further development of the disease.

Usually, when the compositions of the present invention are used for the above purposes, they can be mixed with one or more pharmaceutically acceptable carriers or excipients to make pharmaceutical dosage forms of different administration routes, such as tablets, capsules, powders, Granules, syrups, solutions, oral liquids, spirits, tinctures, aerosols, powder sprays, injections, sterile powders for injections, suppositories, etc.

Usually, the pharmaceutical composition contains: (a) 0.01-99% by weight of the active compound and its pharmaceutically acceptable salt as an active ingredient; (b) a pharmaceutically acceptable carrier. The compounds may also be administered in pure form, or as preparations of various forms of the compounds of the present invention.

A "pharmaceutically acceptable" ingredient is a substance that has a reasonable benefit/risk ratio for use in humans and/or animals without undue adverse side effects (such as toxicity, irritation and allergic reactions). A "pharmaceutically acceptable carrier" is a pharmaceutically or food-acceptable solvent, suspending agent or excipient used to deliver the compound of the present invention or a pharmaceutically acceptable salt thereof to animals or humans. The carrier can be liquid or solid.

The compounds of the present invention can be administered orally, intravenously, intramuscularly or subcutaneously.

Among the above dosage forms, the dosage forms that can be administered orally are: tablets, capsules, powders, granules, syrups, solutions, and spirits. Solid carriers include: starch, lactose, calcium hydrogen phosphate, microcrystalline cellulose, sucrose, kaolin, micronized silica gel, talcum powder, low-substituted hydroxypropyl cellulose, sodium carboxymethyl starch, and polyvinylpyrrolidone. The liquid carrier includes: sterile water, ethanol, polyethylene glycol, nonionic surfactants and edible oils (such as corn oil, peanut oil and sesame oil). Adjuvants commonly used in the process of preparing pharmaceutical compositions include: flavoring agents, coloring agents, preservatives (such as butyl hydroxybenzoate, sodium benzoate, sorbic acid) and antioxidants (such as vitamin E, vitamin C, pyro Sodium Sulfite and Butylated Hydroxytoluene).

Among the above dosage forms, the dosage forms that can be used for administration by injection include: injections and sterile powders for injection, which are prepared by mixing the drug with one or more pharmaceutically acceptable excipients for injection. Solvents include: sterile water, ethanol, glycerin, propylene glycol, polyethylene glycol. In addition, antibacterial agents (such as benzyl alcohol, butylparaben, thimerosal), isotonic regulators (such as sodium chloride, glucose), suspending agents (such as sodium carboxymethylcellulose, methylcellulose, etc.) need to be added. ), solubilizers (Tween-80, lecithin), antioxidants (such as vitamin E, vitamin C, sodium metabisulfite) and fillers (such as lactose, mannitol).

From the standpoint of ease of preparation and administration, preferred pharmaceutical compositions are solid compositions, especially tablets and solid-filled or liquid-filled capsules. Oral administration is preferred.

The effective dose of active ingredient employed will vary with the mode of administration and the severity of the condition being treated. Usually, however, satisfactory results can be obtained when the plant extract of the present invention is administered at a dose of about 0.5-2000 mg/kg of animal body weight per day, preferably in 2-4 divided doses per day, or in slow administered in released form. Dosage forms suitable for internal administration comprise about 0.5-2000 mg of the plant extract in admixture with a solid or liquid pharmaceutically acceptable carrier. This treatment plan can be adjusted to achieve the best therapeutic effect. For example, several divided doses per day may be administered or the dose may be proportionally reduced as indicated by the therapeutic situation. Usually, the oral clinical dose for adults is in the range of 0.5-2000 mg/day, preferably 10-1000 mg/day.

In addition to making medicaments, various food additives such as antioxidants, pigments, and enzyme preparations can also be added to the notoginseng saponin Ft1 of the present invention, and health food can be made according to conventional methods in the field.

Below in conjunction with specific embodiment, further illustrate the present invention. These examples are only for illustrating the present invention and are not intended to limit the scope of the present invention. For the experimental methods without specific conditions indicated in the following examples, usually follow the conventional conditions or the conditions suggested by the manufacturer. All percentages and parts are by weight unless otherwise indicated.

Example 1

1. Experimental materials

1.1 Medicinal materials

Notoginseng saponin Ft1 (HPLC purity ≥98%, Chengdu Master Biotechnology Co., Ltd.); Dextran sodium sulfate (DSS, CAS: 9011-18-1, MW36-50kDa, produced by MP Biomedicals, USA); Willow Azasulfapyridine (SASP, CAS: 599-79-1, molecular weight 398.39, HPLC purity ≥ 98%, produced by Sigma-Aldrich, USA).

1.2 Experimental animals

C57BL/6 female mice (8 weeks) weighing 20±2 g were provided by the Experimental Animal Center of Shanghai University of Traditional Chinese Medicine, animal qualification certificate number: SYXK (Shanghai) 2009-0069. Placed in a conventional feeding environment, free access to food and water.

2. Experimental method

2.1 Establishment of ulcerative colitis model

C57BL/6 female mice (20±2g) with uniform body weight were selected, and the internationally accepted method for modeling ulcerative colitis (Gastroenterology2002, 123:256-70; PLoS One2012, 7:e36075) was used. At the beginning of the study, the Mice had free access to food and water, and after the initial acclimatization phase, the water was changed to 4% (w/v) DSS and drank freely for 7 days to induce ulcerative colitis.

2.2 Grouping and administration method

Normal control group: 10 rats, given normal diet.

Model control group: 10 rats were given 4% DSS solution to drink freely for 7 days.

SASP group: 10 rats were given 4% DSS solution and SASP (350 mg/kg) by intragastric administration for 7 consecutive days.

Notoginseng saponin Ft1 group: 10 rats were administered with 4% DSS solution and intragastrically administered notoginseng saponin Ft1 (50 mg/kg) for 7 consecutive days.

During the experiment, dose formulations were freshly prepared daily, dissolved in 0.5% sodium carboxymethylcellulose, and used within 1 hour of preparation.

2.3 Evaluation of enteritis

Body weight, diarrhea, bloody stool, and histopathological analysis all refer to the methods described in previous research papers (PLoS One2012, 7:e36075; J Pharmacol Exp Ther2013, 345:473-82). Body weight changes, diarrhea and bloody stool symptoms were recorded every day during the experiment. After the experiment, the mice were anesthetized and killed by cervical dislocation, the abdominal cavity was opened, the colon was removed, and the length of the colon from the cecum to the rectum was measured. Tissue sections were taken from the terminal colon, and H&E staining was performed, and pathological scoring was performed on H&E stained colon specimens: (1) Scoring criteria for inflammatory cell exudation: 0 points - very few inflammatory cells in the mucosal lamina propria, 1 point - mucosal lamina propria There are many inflammatory cells in the lamina propria or increased inflammatory cells in the lamina propria, 2 points - inflammatory cells spread to the submucosa, 3 points - inflammatory cells exudate in the whole layer; (2) Scoring criteria for tissue damage: 0 points - no mucosal damage, 1 point - discontinuous mucosal epithelial damage, 2 points - superficial mucosal erosion, 3 points - extensive mucosal damage extending deep into the bowel wall. The histopathological score (1-6 points) was calculated by adding the exudation score of inflammatory cells and the tissue damage score.

2.4 Statistical Analysis

All experimental data were repeated 3 times, and the results were expressed as mean ± standard deviation. SPSS 16.0 statistical software was used to analyze the experimental data by One-way ANOVA and LSD test, and P<0.05 was considered statistically significant. significant.

3. Results

3.1 The therapeutic effect of notoginseng saponin Ft1 on ulcerative colitis

3.1.1 Effects on body weight of mice with ulcerative colitis

Figure 1 shows the effect of notoginseng saponin Ft1 on the body weight of mice with ulcerative colitis. During the whole experiment, the body weight of the mice in the normal control group changed steadily, and the body weight (19.8±0.1g) on the 7th day increased by 0.9%; g) The weight loss rate was 7.9%; the weight of the mice in the SASP group (19.1±0.2g) and the notoginsenoside Ft1 group (18.6±0.2g) continued to drop until the 4th day, and the decline was smaller than that of the model group. The weight loss rates were 2.2% and 4.7%, respectively, which were significantly different from the model group (P<0.001 and P<0.01). It shows that both SASP and notoginseng saponin Ft1 can significantly improve the symptoms of weight loss in mice with ulcerative colitis induced by DSS.

3.1.2 Effects on the incidence of bloody stool in mice with ulcerative colitis

Figure 2 shows the effect of notoginseng saponin Ft1 on the incidence of bloody stool in mice with ulcerative colitis. During the whole experiment, the mice in the normal control group had no bloody stools. From the third day after adding DSS to the drinking water, the mice in the model group and the notoginsenoside Ft1 group began to have bloody stools. The mice in the model group continued to have bloody stools, and the incidence rate of bloody stools was 96.6% on the 7th day; the mice in the SASP group began to have bloody stools on the 4th day when DSS was added to the drinking water, and the mice in the SASP group and notoginsenoside Ft1 group had bloody stools on the 7th day. The incidences of bloody stool in the two days were 36.7% and 40.0%, respectively, which were significantly different from those in the model group (P<0.001 and P<0.01). It shows that both SASP and notoginseng saponin Ft1 can significantly improve the bloody stool symptoms of mice with ulcerative colitis induced by DSS.

3.1.3 Effects on colon length of mice with ulcerative colitis

Figure 3 shows the effect of notoginseng saponin Ft1 on the colon length of mice with ulcerative colitis. Compared with the normal control group (12.0±0.5cm), the colon of the mice in the model group (8.5±0.5cm) was swollen and bleeding (Figure 3A), and was significantly shortened, with a shortening rate of 29.2% (Figure 3B); the SASP group ( 11.2±0.8cm) and notoginsenoside Ft1 group (9.1±0.7cm) mice had slight swelling and hemorrhage in the colon (Fig. 3A), and the shortening rates were 7.1% and 23.6% (Fig. 3B), respectively, compared with the model group There were significant differences (P<0.001 and P<0.05). It shows that both SASP and notoginseng saponin Ft1 can significantly improve the symptoms of colon shortening in mice with ulcerative colitis induced by DSS.

3.1.4 Effects on colon histopathology in mice with ulcerative colitis

Figure 4 shows the damage repair effect of notoginseng saponin Ft1 on the colon tissue of mice with ulcerative colitis. The colon tissue of the mice in the model group showed typical inflammatory mucosal manifestations, with a large number of inflammatory cell infiltration in the mucosa, submucosa and even the muscular layer, tissue structure disorder, rupture, submucosal hemorrhage, edema, and telangiectasia; mice in the SASP group and Panax notoginseng The colon tissue of the mice in the saponin Ft1 group showed that the lesions were alleviated, the ulcers healed, and the submucosa hemorrhage was alleviated (Fig. 4A).

According to the 1-6 point scoring standard representing inflammatory tissue damage, the normal control group scored 0; the model group mice scored (5.7±0.3 points) significantly increased; the SASP group (2.2±0.8 points) The scores of the mice in the saponin Ft1 group (4.1±0.7 points) all decreased, and there were significant differences compared with the model group (P<0.001 and P<0.05) (Figure 4B).

In summary, the results of colon histopathological analysis showed that both SASP and notoginseng saponin Ft1 could significantly improve the histopathological damage of colonic mucosa and the symptoms of inflammatory cell infiltration in mice with ulcerative colitis induced by DSS.

In summary, the results of the study showed that notoginseng saponin Ft1 could significantly improve the symptoms of weight loss, bloody stool, colon shortening and colonic mucosal injury in mice with ulcerative colitis.

The preparation of embodiment 2 tablet

Utilize conventional technique, mix following component, then directly compress tablet, prepare the pharmaceutical composition of tablet form, its formula is as follows:

The scope of the invention is not to be limited by the specific embodiments described, which are intended as single examples illustrating various aspects of the invention, and functionally equivalent methods and components are also intended to be within the scope of the invention. Indeed, various modifications to the invention, in addition to those described herein, can readily be grasped by those skilled in the art upon reference to the foregoing description and accompanying drawings. Such modifications also fall within the scope of the appended claims. Each of the references mentioned above is incorporated herein by reference in its entirety.

Claims (3)

1. Use of notoginsenoside Ft1 or a plant extract containing notoginsenoside Ft1 in the preparation of a medicament for preventing or treating ulcerative colitis. 2. The use according to claim 1, characterized in that the plant extract containing notoginsenoside Ft1 is the ethanol extract of leaves or roots of Araliaceae plants. 3. The use according to claim 2, characterized in that the plant extract containing notoginseng saponin Ft1 is the ethanol extract of Radix Notoginseng.