Application of akebia saponin D in preparation of pharmaceutical composition for preventing and treating constipation

Abstract

The invention relates to the field of pharmaceutical compositions, in particular to application of akebia saponin D or salt thereof in preparation of a pharmaceutical composition for preventing and treating constipation. The pharmaceutical composition containing the akebiaquinata saponin D or the salt of the akebiaquinata saponin D contains the akebiaquinata saponin D or the salt of the akebiaquinata saponin D and pharmaceutic adjuvants. The akebia saponin D can be used for treating functional constipation by increasing the water content of excrement and the number of excrement grains, shortening the defecation time, improving the intestinal propulsion rate and increasing the number of goblet cells.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical compositions, and more particularly to the application of Akebia saponin D in the preparation of pharmaceutical compositions for the prevention and treatment of constipation. Background Technology

[0002] Constipation is a prevalent gastrointestinal dysfunction worldwide, characterized by decreased bowel movement frequency (less than 3 spontaneous bowel movements per week), hard stools, straining during defecation, or a feeling of incomplete evacuation. According to the Rome IV criteria, functional constipation requires exclusion of organic lesions and symptoms persisting for at least 6 months. Epidemiological data shows a global prevalence of approximately 10%-15%, with a significantly higher incidence in women than men (approximately 2.2:1).

[0003] Traditional laxatives (such as rhubarb, mannitol, and polyethylene glycol 4000 powder) only relieve symptoms, and long-term use can easily lead to drug dependence or electrolyte imbalance; gastrointestinal prokinetic drugs (such as domperidone) pose cardiovascular risks.

[0004] Loperamide is a widely used antidiarrheal drug, but it is also a common modeling drug for constipation in laboratory animals. Loperamide acts on μ-opioid receptors in the intestinal wall, inhibiting the release of acetylcholine and prostaglandins from intestinal neurons, thereby weakening the contractile force of intestinal smooth muscle and reducing the frequency of peristalsis. This effect prolongs the retention time of intestinal contents in the intestine, promotes water reabsorption, and leads to hard, dry stools.

[0005] Akebia saponin D, also known as Dipsacus saponin VI, is a triterpenoid saponin compound derived from the dried rhizome of Dipsacus asper. Dipsacus asper has the effects of tonifying the liver and kidneys, strengthening tendons and bones, healing fractures, and stopping metrorrhagia. Akebia saponin D is the main active ingredient in this medicinal herb. Previous studies have shown that Akebia saponin D can improve Alzheimer's disease, diabetic nephropathy, and fractures. Currently, there are no reports on the therapeutic effect of Akebia saponin D on constipation. Summary of the Invention

[0006] The purpose of this invention is to provide the application of akebia saponin D or its salt in the preparation of a pharmaceutical composition for preventing and treating constipation.

[0007] Preferably, the pharmaceutical composition contains a pharmaceutical composition of akebia saponin D or its salt.

[0008] Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0009] Preferably, the pharmaceutically acceptable carrier includes preservatives, antioxidants, flavoring agents, fragrances, solubilizers, emulsifiers, solvents, osmotic pressure regulators, colorants; binders, fillers, disintegrants, and lubricants.

[0010] Preferably, the pharmaceutical composition is prepared into any dosage form by adding pharmaceutically acceptable excipients, including but not limited to tablets, capsules, oral liquids, powders, granules, syrups, solutions, emulsions, suspensions, and suppositories.

[0011] Preferably, the weight content of akebia saponin D or its salt in the drug is 0.1-100%.

[0012] Preferably, the pharmaceutical composition is used in the preparation of a medicament for the prevention and treatment of constipation.

[0013] Preferably, the pharmaceutical composition is used to treat functional constipation. The advantages of this invention are that, after administration of Akebia saponin D, it can treat functional constipation by increasing the number of fecal particles and fecal water content, reducing defecation time, improving intestinal propulsion rate, and increasing the number of goblet cells, and it has good in vivo safety.

[0014] The pharmaceutical compositions or pharmaceutically acceptable compositions described in this invention further comprise pharmaceutically acceptable carriers, including any solvent, diluent, liquid excipient, dispersant, suspending agent, surfactant, isotonic agent, thickener, emulsifier, preservative, solid binder, or lubricant, etc., suitable for a specific target dosage form. Remington: The Science and Practice of Pharmacy, 21st edition, 2005, ed. D.B. Troy, Lippincott Williams & Wilkins, Philadelphia, and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J.C. Boylan, 1988-1999, Marcel Dekker, New York, discloses various carriers used in formulating pharmaceutically acceptable compositions and their known preparation methods. Any other conventional carrier media and their uses are also within the scope of this invention, except for those incompatible with the compounds of this invention, such as those that produce adverse biological effects or interact harmfully with any other component in the pharmaceutically acceptable composition.

[0015] Examples of substances that can be used as pharmaceutically acceptable carriers include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (e.g., human serum albumin), buffers (e.g., Tween 80, phosphates, glycine, sorbic acid, or potassium sorbate), mixtures of metaglycerides of saturated vegetable fatty acids, water, salts, or electrolytes (e.g., protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, or zinc salts), silica gel, magnesium trisilicate, polyvinylpyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-block copolymers, methylcellulose, hydroxypropyl methylcellulose, lanolin, sugars (e.g., lactose, glucose, and sucrose), starches (e.g., corn starch and potato starch), cellulose and its derivatives (e.g., sodium carboxymethyl cellulose, ... Ethyl cellulose and cellulose acetate), powdered tragacanth gum, malt, gel, talc, excipients (e.g., cocoa butter and suppository wax), oils (e.g., peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil), ethylene glycol (e.g., propylene glycol or polyethylene glycol), esters (e.g., ethyl oleate and ethyl dodecanoate), agar, buffers (e.g., magnesium hydroxide and aluminum hydroxide), alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethanol and phosphate buffers, and other non-toxic and compatible lubricants (e.g., sodium lauryl sulfate and magnesium stearate), as well as colorants, anti-sticking agents, coating agents, sweeteners and flavorings, preservatives, and antioxidants, as determined by the formulator, may also be present in the composition.

[0016] The compositions of the present invention can be administered by any suitable means, including oral, rectal, or other routes, of the compounds and pharmaceutically acceptable compositions described above.

[0017] Oral liquid dosage forms include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compound, liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizers, and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butanediol, dimethylformamide, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerin, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, and mixtures thereof. In addition to inert diluents, oral compositions may also include adjuvants, such as humectants, emulsifiers and suspending agents, sweeteners, flavoring agents, and aroma enhancers.

[0018] Oral solid dosage forms include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier, such as sodium citrate or dicalcium phosphate, and / or a) fillers or swelling agents, such as starch, lactose, sucrose, glucose, mannitol, and silica; b) binders, such as carboxymethyl cellulose, alginate, gel, polyvinylpyrrolidone, sucrose, and gum arabic; c) humectants, such as glycerin; d) disintegrants, such as agar, calcium carbonate, potato or cassava starch, alginic acid, certain silicates, and sodium carbonate; e) solution blockers, such as paraffin; f) absorption accelerators, such as quaternary ammonium compounds; g) humectants, such as cetyl alcohol and glyceryl monostearate; h) absorbents, such as kaolin and bentonite; and i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also contain a buffer.

[0019] The active compound may also be in a microencapsulated form having one or more of the excipients described above. Solid dosage forms such as tablets, lozenges, capsules, pills, and granules can be prepared using coatings and shells, such as enteric coatings, controlled-release coatings, and other coatings well known in the pharmaceutical industry. In such solid dosage forms, the active compound may be mixed with at least one inert diluent, such as sucrose, lactose, or starch. Generally, such dosage forms may also contain additional substances besides inert diluents, such as tableting lubricants and other tableting aids, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets, and pills, the dosage form may also contain buffers. They may optionally contain emulsifiers and may also have compositional properties such that the active ingredient is optionally released in a delayed manner only, or preferably, released in a portion of the intestine. Examples of encapsulation compositions that can be used include polymers and waxes.

[0020] The pharmaceutical compositions described herein can also be applied topically, especially when the diseased area or organ (e.g., functional constipation) is easily accessible via local drip application. Suitable local formulations can be readily prepared for each of these areas or organs. Local drip application to the lower intestine can be achieved using rectal suppository formulations or suitable enema formulations.

[0021] Compositions administered rectally are particularly suppositories prepared by mixing the compounds described in this invention with suitable non-irritating excipients or carriers, such as cocoa butter, polyethylene glycol, or suppository waxes, which are solid at ambient temperature but liquid at body temperature and thus melt in the rectum to release the active compound.

[0022] The compounds used in the methods of the present invention can be formulated into unit dosage forms. The term "unit dosage form" refers to a physical discrete unit suitable as a unit dose for a treated patient, each unit containing a predetermined amount of active substance calculated to produce the intended therapeutic effect, optionally bound to a suitable pharmaceutical composition carrier. Unit dosage forms can be used as a single daily dose or as a multiple daily dose (e.g., about 1-4 or more times daily). When using multiple daily doses, the unit dosage form for each dose can be the same or different. Attached Figure Description

[0023] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0024] Figure 1 The effect of constipation model in mice; Figure 2 Different doses of Akebia saponin D were used to treat fecal water content, number of fecal pellets, and time to first black excretion in mice with functional constipation. Figure 3 Intestinal propulsion rate in mice with functional constipation treated with different doses of Akebia saponin D; Figure 4 Results of AB-PAS staining of small intestinal tissue and statistics on the number of goblet cells in a single crypt; Figure 5 Blood biochemical assay of akebia saponin D in mice after a single dose. Detailed Implementation

[0025] The following will describe the concept and technical effects of the present invention clearly and completely with reference to embodiments, so as to fully understand the purpose, features and effects of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are all within the scope of protection of the present invention.

[0026] Example 1: Constipation Mouse Model Laboratory animals: SPF-grade Kunming mice (7 weeks old, female), weighing (45±3) g, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0027] Mice were housed at 24°C with a controlled 12-hour light and 12-hour dark cycle. All mice had free access to food and water, and their bedding was changed every three days. Mice were introduced to the model after one week of acclimatization.

[0028] Animal experimental methods: Mice were divided into 5 groups of 10 mice each, with no significant difference in body weight among the groups. The groups were: blank group, model group, low-dose Akebia saponin D group, medium-dose Akebia saponin D group, and high-dose Akebia saponin D group. The blank group was administered 0.9% saline by gavage daily until the end of the experiment. The model group and each treatment group were administered 0.3 ml of 10 mg / kg bw loperamide hydrochloride by gavage daily. On the 7th day, the stool volume and water content were measured until the model was successfully established. After successful modeling, each treatment group was administered low (75 mg / kg), medium (150 mg / kg), and high (300 mg / kg) doses of Akebia saponin D by gavage once daily, while the model group was administered 0.9% saline by gavage daily for 7 days.

[0029] Example 2: Detection of fecal moisture content, number of fecal pellets excreted within 6 hours, and time of first black pellet excretion. Fecal water content determination: On days 7 and 15, the blank group was given 0.3 mL of 0.9% saline, and the model group and each drug intervention group were given 0.3 mL of loperamide hydrochloride solution (10 mg / kg). One hour later, the mice were placed individually in cages lined with absorbent paper, and feces were collected (only the blank group and the model group were collected on day 7). The wet weight of the feces was obtained by weighing, and the dry weight of the feces was obtained by lyophilizing. The fecal water content was calculated according to formula (1):

[0030] Number of fecal pellets in 6 hours: On days 7 and 15, 1 hour after gavage, individual mice were placed in cages lined with absorbent paper and feces were collected every 30 minutes (only the control group and the model group were collected on day 7), for a total of 6 hours.

[0031] Time of first black stool expulsion: On day 15, 1 hour after gavage, each group was gavage with 0.2 mL of 5% activated charcoal (prepared with 0.2% sodium carboxymethyl cellulose) solution. The time of first black stool expulsion for each mouse was recorded from the time of gavage with activated charcoal.

[0032] like Figure 1 The results showed that 7 days after modeling, the fecal water content and fecal particle number in the blank group were 1.28 times and 3.89 times that in the model group, respectively, and both were significantly different, indicating that the constipation model was successfully established.

[0033] like Figure 2The results showed that after 7 days of treatment with different doses of Akebia saponin D, low-dose (75 mg / kg), medium-dose (150 mg / kg), and high-dose (300 mg / kg) Akebia saponin D improved the time to first black stool expulsion, number of fecal particles, and fecal water content in a dose-dependent manner compared to the model group. The high-dose (300 mg / kg) Akebia saponin D showed the best therapeutic effect, significantly increasing fecal water content by 1.33 times, significantly increasing fecal particle number by 2.47 times, and significantly reducing the time to first black stool expulsion by 70 minutes compared to the model group.

[0034] Example 3: Determination of intestinal propulsion rate in mice On day 13, five mice from each group were fasted but allowed free water overnight. On day 14, at 8:00 AM, the blank control group was given 0.3 mL of 0.9% saline solution, and the model group and each drug intervention group were given 0.3 mL of loperamide hydrochloride solution (10 mg / kg). 30 minutes later, the blank control group and the model group were given 0.2 mL of 5% activated charcoal solution by gavage, and each drug intervention group was given activated charcoal solution containing their respective concentrations of akebia saponin D by gavage. 30 minutes later, the mice were sacrificed, the abdominal cavity was opened, and the upper end from the pylorus to the cecum was cut off. The total length of the intestine was measured as the "total intestinal length", and the length from the pylorus to the front of the activated charcoal was measured as the "activated charcoal propulsion length". The intestinal propulsion rate was calculated according to formula (2):

[0035] like Figure 3 The results showed that the intestinal propulsion rate of mice was improved after treatment with different doses of Akebia saponin D, with the high dose of Akebia saponin D (300 mg / kg) significantly increasing it by 1.28 times compared with the model group.

[0036] Example 4: AB-PAS staining analysis of small intestinal tissue to determine the number of goblet cells in a single crypt. On day 16, mice in all groups were fasted but allowed free access to water overnight. At 8:00 AM on day 17, the control group was given 0.3 mL of 0.9% saline solution, while the model group and all drug intervention groups were given 0.3 mL of loperamide hydrochloride solution (10 mg / kg bw). Thirty minutes later, the mice were euthanized by cervical dislocation, and small intestinal tissue was removed and fixed in 4% paraformaldehyde. After dehydration, embedding, and paraffin sectioning, the paraffin sections were dewaxed to water. The dewaxed sections were stained in AB-PAS staining solution C for 15 min, then washed with tap water until colorless. The sections were then immersed in AB-PAS staining solution B for 15 min, washed with tap water, and then twice with distilled water. The sections were then stained in AB-PAS staining solution A (at room temperature) in the dark for 30 min, followed by rinsing with running water for 5 min. Finally, the sections were dehydrated, mounted, and images were collected under a microscope to analyze the number of goblet cells in individual crypts.

[0037] like Figure 4 The results showed that akebia saponin D could increase the number of goblet cells in a single crypt in the small intestine in a dose-dependent manner.

[0038] Example 5: Toxicity test of Akebia saponin D in mice after a single dose Sixty SPF-grade Kunming mice (7 weeks old, female), weighing (45±3) g, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. The mice were randomly divided into four groups of 15 mice each, and their weight was recorded. The groups included a control group administered saline by gavage, and three groups administered Akebia saponin D by gavage (dose of 500, 2000, and 5000 mg / kg, respectively). Mice were fasted for 10 hours before administration and for another 4 hours after administration. They were then observed for 14 consecutive days. General physical signs, behavior, mental status, and mortality were observed and recorded daily. On days 1, 7, and 14 after administration, serum was collected from 5 mice in each group for blood biochemical analysis, including alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), creatinine, total bilirubin, urea, and creatine kinase.

[0039] like Figure 5 The results showed that high doses of Akebia saponin D had no significant effect on alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), creatinine, total bilirubin, urea, and creatine kinase. Furthermore, mice in all groups maintained normal coat color and showed no hair follicle protrusion. This indicates that Akebia saponin D has good in vivo safety.

[0040] In summary, Akebia saponin D can treat functional constipation by increasing fecal water content and fecal particle number, reducing defecation time, improving intestinal propulsion rate, and increasing goblet cell number, and it has good in vivo safety.

[0041] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. Use of Akebia saponin D or its salt in the preparation of pharmaceutical compositions for the prevention and treatment of constipation.

2. The use of akebia saponin D or its salt according to claim 1 in the preparation of a pharmaceutical composition for preventing and treating constipation, characterized in that, The pharmaceutical composition comprises akebia saponin D or its salt and a pharmaceutically acceptable carrier.

3. The use according to claim 2, wherein the pharmaceutical composition includes, but is not limited to, tablets, capsules, oral liquids, powders, granules, syrups, solutions, emulsions, suspensions, and suppositories.

4. The use according to claim 2, characterized in that, The weight content of akebia saponin D or its salt in the pharmaceutical composition is 0.1-100%.

5. The use according to claim 2, characterized in that, Pharmaceutically acceptable carriers include preservatives, antioxidants, flavoring agents, fragrances, solubilizers, emulsifiers, solvents, osmotic pressure regulators, colorants; binders, fillers, disintegrants, and lubricants.

6. The use according to any one of claims 1-5, characterized in that, The pharmaceutical composition treats functional constipation by increasing the number of fecal particles and the water content of feces, reducing defecation time, improving intestinal propulsion rate, and increasing the number of goblet cells, and has good in vivo safety.

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