A suppository containing pomegranate flowers, its preparation method and uses

Abstract

This invention discloses a suppository containing pomegranate flowers. The suppository comprises catechu, alum, gallnut, pomegranate peel, pomegranate flowers, a first excipient, and a second excipient. The first excipient includes polyethylene glycol and polyoxyethylene (40) monostearate, and the second excipient is glycerin. The suppository prepared according to this invention has a simple and convenient process and a high content of active ingredients. It can be used for the prevention and / or treatment of nonspecific vaginitis, such as fungal vaginitis, and exerts anti-inflammatory and antipruritic effects.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to a suppository containing pomegranate flowers, its preparation method, and its uses. Background Technology

[0002] Nonspecific vaginitis, such as yeast infection, is a common and frequently occurring inflammatory disease of the vulva and vagina, mainly caused by Candida infection. Symptoms often include increased vaginal discharge and vulvar itching, significantly impacting the patient's quality of life. Currently, there are many clinical treatment options for yeast infections, often using antifungal drugs to effectively inhibit the pathogen and achieve therapeutic goals. However, practice has shown that this treatment method has certain side effects, affecting the normal flora and disrupting the vaginal microecological environment.

[0003] There are no existing reports in the art regarding the use of a suppository containing pomegranate flowers, as claimed in this invention, for the prevention and / or treatment of nonspecific vaginitis, such as fungal vaginitis. Summary of the Invention

[0004] Based on this, the present invention provides a suppository containing pomegranate flowers, the suppository comprising catechu, alum, gallnut, pomegranate peel, pomegranate flowers, a first excipient and a second excipient, wherein the first excipient comprises polyethylene glycol and polyoxyethylene (40) monostearate, and the second excipient is glycerin.

[0005] Furthermore, the polyethylene glycol is selected from one or more of the following: polyethylene glycol 400, polyethylene glycol 4000, and polyethylene glycol 6000.

[0006] Furthermore, the mass ratio of the catechu, the alum, the gallnut, the pomegranate peel, the pomegranate flower, the first excipient, and the second excipient is (2-4):(0.5-2):(2-4):(1-3):(1-3):(20-30):(2-8).

[0007] Furthermore, the mass ratio of the catechu, the alum, the gallnut, the pomegranate peel, the pomegranate flower, the first excipient, and the second excipient is approximately 3.15: approximately 1.26: approximately 3.15: approximately 1.89: approximately 1.89: approximately 25.1: approximately 5.58.

[0008] Furthermore, the mass ratio of the polyethylene glycol 400, the polyethylene glycol 4000, the polyethylene glycol 6000 and the polyoxyethylene (40) monostearate is (2.5-3.5):(0.5-1.5):(1.5-2.5):(1.5-2.5).

[0009] Furthermore, the mass ratio of the polyethylene glycol 400, the polyethylene glycol 4000, the polyethylene glycol 6000 and the polyoxyethylene (40) monostearate is about 3: about 1: about 2: about 2.

[0010] Furthermore, the suppository further comprises one or more drugs and / or extracts for the prevention and / or treatment of nonspecific vaginitis.

[0011] Furthermore, the drug is selected from one or more of the following: metronidazole, clindamycin, fluconazole, miconazole, thiamphenicol, mepramycin, clotrimazole, nystatin, 10% Jieeryin lotion and Jieeryin effervescent tablets.

[0012] According to another aspect of the present invention, a method for preparing the suppository is provided, the method comprising the following steps:

[0013] (1) Weigh out an appropriate amount of catechu and alum, mix them and grind them into fine powder, and optionally sieve them to obtain the first mixture;

[0014] (2) Weigh out appropriate amounts of gallnut, pomegranate peel and pomegranate flower, mix them and crush them into coarse powder, add solvent to extract, centrifuge to take the supernatant, concentrate, dry, crush into fine powder, and optionally sieve to obtain the second mixture;

[0015] (3) Mix the first mixture with the second mixture to obtain a third mixture;

[0016] (4) Weigh appropriate amounts of the first and second excipients, melt them in a water bath, add the third mixture while stirring in the water bath, mix thoroughly to obtain the fourth mixture; and

[0017] (5) The fourth mixture is injected into a suppository mold, cooled, and granulated to obtain the suppository.

[0018] Furthermore, in step (2), the solvent is water or ethanol.

[0019] Furthermore, the solvent is water.

[0020] Furthermore, the volume of the solvent is 6 to 14 times the weight of the coarse powder, for example, about 10 times.

[0021] Furthermore, the extraction method is heating and reflux extraction.

[0022] Furthermore, the temperature of the heating reflux extraction is 50°C to 100°C, for example, 100°C.

[0023] Furthermore, the extraction can be performed 1 to 5 times, for example, 3 times.

[0024] Furthermore, the extraction time is 1 hour / time to 2 hours / time, for example, about 1.5 hours / time.

[0025] Furthermore, the centrifugation speed is 1000 r / min to 3000 r / min, for example, about 1500 r / min.

[0026] Furthermore, the centrifugation time is 8 to 12 minutes, for example, about 10 minutes.

[0027] Furthermore, this concentration is a vacuum concentration.

[0028] Furthermore, the pressure of the vacuum concentration is 0.06 MPa to 1.00 MPa, for example, about 0.08 MPa.

[0029] Furthermore, the temperature for this vacuum concentration is 45°C to 75°C, for example, about 50°C.

[0030] Furthermore, the supernatant is concentrated under reduced pressure to a relative density of 1.10–1.30, for example, 1.15–1.20, at 60°C.

[0031] Furthermore, the drying process is vacuum drying.

[0032] Furthermore, the drying pressure is 0.06 MPa to 1.00 MPa, for example, about 0.08 MPa.

[0033] Furthermore, the drying temperature is 55°C to 75°C, for example, about 70°C.

[0034] Furthermore, in steps (1) and (2), the sieve is a 100-mesh sieve.

[0035] Further, in step (4), the first excipient includes polyethylene glycol and polyoxyethylene (40) monostearate, and the second excipient is glycerin.

[0036] Furthermore, the polyethylene glycol is selected from one or more of the following: polyethylene glycol 400, polyethylene glycol 4000, and polyethylene glycol 6000.

[0037] Furthermore, the mass ratio of the polyethylene glycol 400, the polyethylene glycol 4000, the polyethylene glycol 6000 and the polyoxyethylene (40) monostearate is (2.5-3.5):(0.5-1.5):(1.5-2.5):(1.5-2.5).

[0038] Furthermore, the mass ratio of the polyethylene glycol 400, the polyethylene glycol 4000, the polyethylene glycol 6000 and the polyoxyethylene (40) monostearate is about 3: about 1: about 2: about 2.

[0039] Furthermore, the water bath temperature for melting is 60℃~70℃.

[0040] Furthermore, the water bath temperature for stirring is 60℃~65℃.

[0041] Furthermore, the stirring time should be at least 20 minutes.

[0042] Furthermore, in step (5), the mass of the suppository is approximately 3.0 g.

[0043] Furthermore, the suppository has a duckbill-shaped mold.

[0044] According to another aspect of the invention, the use of the above-described suppository in the preparation of a medicament for the prevention and / or treatment of nonspecific vaginitis is provided.

[0045] Furthermore, this nonspecific vaginitis is fungal vaginitis.

[0046] According to another aspect of the invention, the use of the above-described suppository in the preparation of a medicament for relieving inflammatory swelling is provided.

[0047] According to another aspect of the invention, the use of the above-described suppository in the preparation of a medicament for inhibiting inflammatory exudation is provided.

[0048] According to another aspect of the invention, the use of the above-described suppository in the preparation of a medicament for the prevention and / or treatment of pruritus is provided.

[0049] The beneficial effects of this invention are:

[0050] The suppository preparation process of the present invention is simple, convenient, and has a high content of active ingredients. It can be used for the prevention and / or treatment of nonspecific vaginitis, such as fungal vaginitis, and to exert anti-inflammatory and antipruritic effects. Attached Figure Description

[0051] To more clearly illustrate the technical solutions in the embodiments of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without exceeding the scope of protection claimed by the present invention.

[0052] Figure 1 This is a flow chart of the suppository manufacturing process of the present invention. Detailed Implementation

[0053] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. 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.

[0054] Unless otherwise stated, all technical and scientific terms and abbreviations used herein have the meanings commonly understood by one of ordinary skill in the field of this invention or the field of application of such terms. While any methods, conditions, substances, or materials similar to or equivalent to those disclosed herein may be used in the practice of this invention, preferred methods, conditions, substances, or materials are described herein.

[0055] This invention is intended to cover all options, variations, and equivalents that may be included in the field of prior art as defined in the claims. Those skilled in the art will recognize many similar or equivalent methods and substances described herein that can be applied in the practice of this invention. This invention is by no means limited to the description of methods and substances.

[0056] The singular forms “a,” “an,” and “the” used in the specification and appended claims include plural indicators unless the context clearly specifies otherwise.

[0057] In this invention, the term "comprising" and "including" are synonymous. The terms "comprising," "including," "having," "containing," or any other variations thereof as used herein are intended to cover a non-exclusive inclusion. For example, a composition, step, method, article, or apparatus that includes the listed elements is not necessarily limited to those elements, but may include other elements not expressly listed or elements inherent to such a composition, step, method, article, or apparatus.

[0058] As described in the background section, the available clinical treatments for fungal vaginitis have certain side effects, and there are no prior art reports on the use of the pomegranate flower-containing suppository claimed in this invention for the prevention and / or treatment of nonspecific vaginitis such as fungal vaginitis. To address the above problems, this invention provides a pomegranate flower-containing suppository comprising catechu, alum, gallnut, pomegranate peel, pomegranate flower, a first excipient, and a second excipient, wherein the first excipient comprises polyethylene glycol and polyoxyethylene (40) monostearate, and the second excipient is glycerin.

[0059] This invention utilizes modern advanced pharmaceutical technology and production processes to produce "Pomegranate Flower Suppository," a specialized preparation of Uyghur medicine suppositories for gynecological diseases. This preparation features a refined formula, outstanding efficacy, distinctive characteristics, a rational dosage form, and convenient use. The successful development of this product will provide a wide range of female patients with a highly effective, high-quality, safe, non-toxic, simple, and inexpensive new Uyghur medicine. It will undoubtedly make a positive contribution to the modernization of ethnic medicine and create significant social and economic benefits.

[0060] In a preferred embodiment, the polyethylene glycol is selected from one or more of the following: polyethylene glycol 400, polyethylene glycol 4000, and polyethylene glycol 6000.

[0061] In a preferred embodiment, the mass ratio of the catechu, the alum, the gallnut, the pomegranate peel, the pomegranate flower, the first excipient, and the second excipient is (2-4):(0.5-2):(2-4):(1-3):(1-3):(20-30):(2-8).

[0062] In this invention, when a mass ratio, weight, temperature, number of times, rotation speed, relative density, time, pressure, proportion, equivalent, concentration, or other value or parameter is expressed as a range, preferred range, or a range defined by a series of upper and lower preferred values, this should be understood as specifically disclosing all ranges formed by any pairing of any upper or preferred value with any lower or preferred value, regardless of whether the range is disclosed individually. For example, when the range "20-30" is disclosed, the described range should be interpreted as including ranges "20-30", "20-29", "20-28", "20-27", "20-26", "20-25", "20-24", "20-23", "20-22", "20-21", "21-30", "21-29", "21-28", "21-27", "21-26", "21-25", "21-24", "21-23", "21-22", "22-30", etc. When a range of values ​​is described herein, unless otherwise stated, the range is intended to include its endpoints and all integers and fractions within that range, and the technical effects of the present invention can be achieved within the aforementioned range.

[0063] In a preferred embodiment, the mass ratio of the catechu, the alum, the gallnut, the pomegranate peel, the pomegranate flower, the first excipient, and the second excipient is about 3.15: about 1.26: about 3.15: about 1.89: about 1.89: about 25.1: about 5.58.

[0064] In this invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 3.15" includes ±5% of 3.15, or from 2.9925 to 3.3075; "about 1.26" includes ±5% of 1.26, or from 1.197 to 1.323; "about 1.89" includes ±5% of 1.89, or from 1.7955 to 1.9845; "about 25.1" includes ±5% of 25.1, or from 23.845 to 26.355; and "about 5.58" includes ±5% of 5.58, or from 5.301 to 5.859.

[0065] In a preferred embodiment, the mass ratio of polyethylene glycol 400, polyethylene glycol 4000, polyethylene glycol 6000 and polyoxyethylene (40) monostearate is (2.5-3.5):(0.5-1.5):(1.5-2.5):(1.5-2.5).

[0066] In a preferred embodiment, the mass ratio of the polyethylene glycol 400, the polyethylene glycol 4000, the polyethylene glycol 6000 and the polyoxyethylene (40) monostearate is about 3: about 1: about 2: about 2.

[0067] In this invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 3" includes 3 ±5%, or from 2.85 to 3.15; "about 1" includes 1 ±5%, or from 0.95 to 1.05; and "about 2" includes 2 ±5%, or from 1.9 to 2.1.

[0068] In a preferred embodiment, the suppository further comprises one or more drugs and / or extracts for the prevention and / or treatment of nonspecific vaginitis.

[0069] In a preferred embodiment, the drug is selected from one or more of the following: metronidazole, clindamycin, fluconazole, miconazole, thiamphenicol, metoprolol, clotrimazole, nystatin, 10% Jieeryin lotion and Jieeryin effervescent tablets.

[0070] According to another aspect of the present invention, a method for preparing the suppository is provided, the method comprising the following steps:

[0071] (1) Weigh out an appropriate amount of catechu and alum, mix them and grind them into fine powder, and optionally sieve them to obtain the first mixture;

[0072] (2) Weigh out appropriate amounts of gallnut, pomegranate peel and pomegranate flower, mix them and crush them into coarse powder, add solvent to extract, centrifuge to take the supernatant, concentrate, dry, crush into fine powder, and optionally sieve to obtain the second mixture;

[0073] (3) Mix the first mixture with the second mixture to obtain a third mixture;

[0074] (4) Weigh appropriate amounts of the first and second excipients, melt them in a water bath, add the third mixture while stirring in the water bath, mix thoroughly to obtain the fourth mixture; and

[0075] (5) The fourth mixture is injected into a suppository mold, cooled, and granulated to obtain the suppository.

[0076] In a preferred embodiment, in step (2), the solvent is water or ethanol.

[0077] In a preferred embodiment, the solvent is water.

[0078] In a preferred embodiment, the volume of the solvent is 6 to 14 times the weight of the coarse powder, for example, about 10 times.

[0079] In this invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 10" includes ±5% of 10, or from 9.5 to 10.5.

[0080] In a preferred embodiment, the extraction method is heating and reflux extraction.

[0081] In a preferred embodiment, the temperature of the heating reflux extraction is 50°C to 100°C, for example, 100°C.

[0082] In this invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 100" includes ±5% of 100, or from 95 to 105.

[0083] In a preferred embodiment, the extraction is performed 1 to 5 times, for example, 3 times.

[0084] In a preferred embodiment, the extraction time is 1 hour / time to 2 hours / time, for example, about 1.5 hours / time.

[0085] In this invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 1.5" includes ±5% of 1.5, or from 1.425 to 1.575.

[0086] In a preferred embodiment, the centrifugation speed is 1000 r / min to 3000 r / min, for example, about 1500 r / min.

[0087] In this invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 1500" includes ±5% of 1500, or from 1425 to 1575.

[0088] In a preferred embodiment, the centrifugation time is 8 min to 12 min, for example, about 10 min.

[0089] In this invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 10" includes ±5% of 10, or from 9.5 to 10.5.

[0090] In a preferred embodiment, the concentration is a vacuum concentration.

[0091] In a preferred embodiment, the pressure of the vacuum concentration is 0.06 MPa to 1.00 MPa, for example, about 0.08 MPa.

[0092] In this invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 0.08" includes ±5% of 0.08, or from 0.076 to 0.084.

[0093] In a preferred embodiment, the temperature for vacuum concentration is 45°C to 75°C, for example, about 50°C.

[0094] In this invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 50" includes 50 ± 5%, or from 47.5 to 52.5.

[0095] In a preferred embodiment, the supernatant is concentrated under reduced pressure to a relative density of 1.10 to 1.30, for example, 1.15 to 1.20, at 60°C.

[0096] In a preferred embodiment, the drying is vacuum drying.

[0097] In a preferred embodiment, the drying pressure is 0.06 MPa to 1.00 MPa, for example, about 0.08 MPa.

[0098] In this invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 0.08" includes ±5% of 0.08, or from 0.076 to 0.084.

[0099] In a preferred embodiment, the drying temperature is 55°C to 75°C, for example, about 70°C.

[0100] In this invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 70" includes 70 ±5%, or from 66.5 to 73.5.

[0101] In a preferred embodiment, in steps (1) and (2), the sieve is a 100-mesh sieve.

[0102] In a preferred embodiment, in step (4), the first excipient comprises polyethylene glycol and polyoxyethylene (40) monostearate, and the second excipient is glycerin.

[0103] In a preferred embodiment, the polyethylene glycol is selected from one or more of the following: polyethylene glycol 400, polyethylene glycol 4000, and polyethylene glycol 6000.

[0104] In a preferred embodiment, the mass ratio of polyethylene glycol 400, polyethylene glycol 4000, polyethylene glycol 6000 and polyoxyethylene (40) monostearate is (2.5-3.5):(0.5-1.5):(1.5-2.5):(1.5-2.5).

[0105] In a preferred embodiment, the mass ratio of the polyethylene glycol 400, the polyethylene glycol 4000, the polyethylene glycol 6000 and the polyoxyethylene (40) monostearate is about 3: about 1: about 2: about 2.

[0106] In this invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 3" includes 3 ±5%, or from 2.85 to 3.15; "about 1" includes 1 ±5%, or from 0.95 to 1.05; and "about 2" includes 2 ±5%, or from 1.9 to 2.1.

[0107] In a preferred embodiment, the melting water bath temperature is 60°C to 70°C.

[0108] In a preferred embodiment, the water bath temperature for stirring is 60°C to 65°C.

[0109] In a preferred embodiment, the stirring time is at least 20 minutes.

[0110] In a preferred embodiment, in step (5), the mass of the suppository is about 3.0g.

[0111] In this invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 3" includes 3 ± 5%, or from 2.85 to 3.15.

[0112] In a preferred embodiment, the suppository has a duckbill shape.

[0113] According to another aspect of the invention, the use of the above-described suppository in the preparation of a medicament for the prevention and / or treatment of nonspecific vaginitis is provided.

[0114] In a preferred embodiment, the nonspecific vaginitis is fungal vaginitis.

[0115] According to another aspect of the invention, the use of the above-described suppository in the preparation of a medicament for relieving inflammatory swelling is provided.

[0116] According to another aspect of the invention, the use of the above-described suppository in the preparation of a medicament for inhibiting inflammatory exudation is provided.

[0117] According to another aspect of the invention, the use of the above-described suppository in the preparation of a medicament for the prevention and / or treatment of pruritus is provided.

[0118] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions or conditions recommended by the manufacturer.

[0119] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to the methods of this invention. The preferred embodiments and materials described herein are for illustrative purposes only.

[0120] The features mentioned above in this invention, or the features mentioned in the embodiments, can be combined arbitrarily. All features disclosed in this patent specification can be used in any compositional form, and each feature disclosed in the specification can be replaced by any alternative feature that provides the same, equivalent, or similar purpose. Therefore, unless otherwise specified, the disclosed features are merely general examples of equivalent or similar features.

[0121] Example

[0122] Preparation Examples

[0123] The conventional process for the pharmaceutical composition of this invention involves water extraction of pomegranate peel, followed by pulverizing the remaining four medicinal materials for use in the medicine. To fully consider the medicinal use of gallnut and pomegranate flower, a comparative study of two extraction processes, a new one and the old one, was designed. The specific schemes are as follows:

[0124] Traditional process: Pomegranate peel is extracted separately with water.

[0125] New process: Extraction of gallnut, pomegranate peel and pomegranate flower together.

[0126] After optimizing both extraction processes, samples from both processes were compared. Based on the comparison results, the optimal extraction process was selected, along with the pulverized medicinal materials used in suppository preparation. Among these, pomegranate flower has a weak chemical research foundation, mainly containing tannins. In the original process, gallnut and pomegranate flower were added after pulverization. This invention focuses on the extraction process based on the chemical composition of the medicine and the safety of local administration. Single-factor and orthogonal experiments will be used to study the extraction processes of gallnut, pomegranate peel, and pomegranate flower to select the best extraction process.

[0127] 1. Water absorption rate test

[0128] Weigh out 1 / 10 of the prescription ratio of coarse powder of gallnut, pomegranate peel, and pomegranate flower, add 6 times the amount of water to soak until thoroughly soaked, filter, and the water absorption rate is measured to be 127.22%.

[0129] 2. Extraction process research

[0130] Single-factor and orthogonal experiments were used to investigate the factors affecting the experimental results, including extraction method, extraction temperature, extraction solvent, water addition, number of extractions, and extraction time, and to select the optimal extraction process.

[0131] 2.1 Evaluation Indicators

[0132] This study uses a comprehensive score as the evaluation index, as follows: Since gallnut is one of the main ingredients in the prescription, primarily containing tannin derivatives, and pomegranate peel and flowers mainly contain tannins and ellagic acid, the contents of tannins, gallic acid, and ellagic acid, as well as the amount of water extract, are used as evaluation indicators, with weighting coefficients set at 0.2, 0.3, 0.3, and 0.2, respectively. The highest score for each indicator is set at 100 points, and each indicator is multiplied by its weighting coefficient to obtain its score. The sum of the scores for all four indicators is the comprehensive score.

[0133] 2.1.1 The gallic acid content was determined by high performance liquid chromatography (HPLC) according to the method described in the 2015 edition of the Chinese Pharmacopoeia, Part IV, 0512.

[0134] Chromatographic conditions and system suitability test: Waters XBridge Shield RP C 18 (4.6×250mm, 5μm) chromatographic column. Mobile phase: acetonitrile (A)-0.2% formic acid aqueous solution (B) gradient elution: 0–9 min (2%–2% A), 9–10 min (2%–24% A), 10–25 min (24%–24% A); flow rate: 1.0 ml / min; column temperature: 35℃; detection wavelength: 254 nm; theoretical plate count based on gallic acid should be no less than 3000.

[0135] Preparation of reference solutions: Accurately weigh an appropriate amount of gallic acid reference standard and add methanol to prepare a solution containing 0.476 mg per ml. Accurately weigh an appropriate amount of ellagic acid reference standard and add methanol to prepare a solution containing 0.604 mg per ml.

[0136] Preparation of the test solution: Accurately measure 1 ml of sample solution, add methanol to make up to 50 ml, sonicate for 10 min, filter through a microporous membrane (0.45 μm), discard the initial filtrate, and take the subsequent filtrate as the test solution.

[0137] The assay involves precisely pipetting 10 μL each of the reference solution and the test solution into a liquid chromatograph and measuring the results.

[0138] 2.1.2 Determination of Total Tannin Content: Follow the method for determination of tannin content (Chinese Pharmacopoeia 2015 Edition, Part IV, 2202).

[0139] (1) Preparation of reference solution

[0140] Accurately weigh 25.3 mg of gallic acid reference standard and place it in a 50 ml brown volumetric flask. Dilute with water to the mark. Then accurately measure 5 ml of the solution and place it in a 50 ml brown volumetric flask. Dilute with water to the mark to obtain a 0.0506 mg / ml gallic acid reference standard solution.

[0141] (2) Preparation of standard curve

[0142] Accurately measure 0.5, 1, 2, 3, 4, 5, and 6 ml of the reference solution and place them in 25 ml brown volumetric flasks respectively. Add 1 ml of phosphomolybdic tungstic acid test solution to each flask, and then add 11.5, 11, 10, 9, 8, 7, and 6 ml of water respectively. Dilute to the mark with 29% sodium carbonate solution, shake well, and let stand for 30 min. Using the corresponding reagent as a blank, measure the absorbance at a wavelength of 760 nm according to the ultraviolet spectrophotometry method (Appendix VA). Plot a standard curve with absorbance as the ordinate and concentration as the abscissa.

[0143] (3) Preparation of the test solution

[0144] Take 1 / 10 of the prescription ratio of gallnut, pomegranate peel and pomegranate flower, grind them through a 60-mesh sieve, put the fine powder into a 1000ml volumetric flask, add 700ml of water and soak overnight, then make up to 1000ml with water, filter, take 2ml of the filtrate and make up to 100ml with water, then take 5ml of the filtrate and make up to 50ml with water, set aside.

[0145] (4) Determination of total phenols

[0146] Accurately measure 2 ml of the test solution and place it in a 25 ml brown volumetric flask. Following the method under the preparation section of the standard curve, starting from "add 1 ml of phosphomolybdic tungstic acid test solution", add 10 ml of water and determine the absorbance according to the method. Calculate the total phenol content (mg).

[0147] (5) Polyphenols that are not absorbed

[0148] Accurately measure 25 ml of the test solution and add it to a 100 ml stoppered conical flask containing 0.6 g of casein. Seal the flask tightly and incubate it in a 30 °C water bath for 1 h, shaking occasionally. Remove the flask, cool it, shake it well, filter it, and discard the initial filtrate. Accurately measure 2 ml of the subsequent filtrate and place it in a 25 ml brown volumetric flask. Following the method under the preparation of the standard curve, starting from "adding 1 ml of phosphomolybdic tungstic acid test solution", add 10 ml of water. Measure the absorbance using the pre-determined method and calculate the total phenol content (mg) that is not absorbed.

[0149] The tannin content is calculated using the following formula:

[0150] Tannin content = (Total phenol content - Non-absorbed polyphenol content) / Medicinal material content (mg / g)

[0151] 2.1.3 Determination of water leachable content

[0152] Accurately measure 50 ml of the extract and place it in an evaporating dish that has been dried to constant weight. Evaporate to dryness on a water bath, then dry at 105°C for 3 hours. Cool in a desiccator for 30 minutes, and quickly and accurately weigh the extract to calculate the amount of extract.

[0153] 2.2 Single-factor experiment

[0154] The main factors affecting water extraction include the influence of different solvents on the active ingredients of medicinal materials; comparative analysis of extraction processes of single-herb decoction and mixed decoction of medicinal materials; investigation of the influence of temperature on extraction process of different solvents of medicinal materials; water absorption rate test; orthogonal experimental study on each level of these influencing factors, including water addition, decoction time and number of extractions, using single-factor analysis; and verification experiment.

[0155] 2.2.1 Investigation of Extraction Solvent Types

[0156] Weigh out 4 portions of pomegranate peel, pomegranate flower, and coarse gallic acid powder according to 1 / 10 of the prescription amount. Extract once, each time for 1.0 hour, with the solvent volume being 10 times. The extraction solvents are water, 30%, 50%, and 70% ethanol, respectively. The yield of extract, tannin, gallic acid, and ellagic acid content are determined and analyzed. The results are shown in Table 1.

[0157] Table 1. Extraction results with different solvents

[0158] solvent Overall score water 100.00 30% ethanol 61.86 50% ethanol 61.12 70% ethanol 64.77

[0159] The results above show that water extraction yielded the highest overall extraction rate for the three medicinal materials, therefore water was chosen as the extraction solvent.

[0160] 2.2.2 Investigation of Individual and Mixed Extraction Processes of Medicinal Herbs

[0161] Two extraction methods were used for the medicinal materials in the prescription: single extraction and mixed extraction. The contents of gallic acid and tannins and the yield of extract were compared. Gallic fruit, pomegranate peel and pomegranate flower were weighed according to the prescription ratio, and 8 times the amount of water was added to each. Extraction was performed three times, for 2, 1 and 1 hours respectively. After filtration, the total volume of the filtrates was measured, and the yield of extract, the contents of tannins and gallic acid were determined. A comprehensive score was also calculated. The results are shown in Table 2.

[0162] Table 2 Extraction results of single and combined decoctions of medicinal materials

[0163] sample Overall score Artemisia 88.31 pomegranate peel 40.96 pomegranate flowers 37.90 mix 73.50

[0164] The results above show that the overall extraction rate of mixed extraction is higher than that of individual extraction, therefore, mixed extraction is the preferred method for the three medicinal materials.

[0165] 2.2.3 Investigation of different extraction temperatures

[0166] According to the prescription, a total of 69.5g of pomegranate peel, pomegranate flowers, and gallic acid were weighed into 5 portions. The extracts were extracted three times with ten times the amount of water, each time for 1 hour. The extraction temperatures were 25, 50, 70, 90, and 100℃. The yield of extract, tannin content, and gallic acid content were determined, and a comprehensive score was given. The results are shown in Table 3.

[0167] Table 3 Results of the extraction temperature study

[0168] Extraction temperature (°C) Overall score 25 34.62 50 44.55 80 55.54 100 99.76

[0169] As can be seen from the above results, the overall extraction rate obtained by boiling and reflux extraction is the highest. Therefore, the three medicinal materials were selected for mixed reflux extraction.

[0170] 2.2.4 Water addition assessment

[0171] Weigh out 1 / 10 of the prescription medicinal materials, namely 31.50g of gallnut, 18.90g of pomegranate peel, and 18.90g of pomegranate flower, in 5 portions each, and place them in 2000ml round-bottom flasks. Add 6, 8, 10, 12, and 14 times the amount of water respectively, reflux for 1 hour, and extract 3 times. Filter, combine the filtrates, cool to room temperature, measure the total volume, and determine the yield of extract, the content of tannins, gallic acid, and ellagic acid. A comprehensive score is then given, and the results are shown in Table 4.

[0172] Table 4 Results of the investigation on the amount of water added

[0173] Add water times (times) Overall score 6 81.08 10 94.10 14 90.10 18 87.90

[0174] The results showed that adding 10 units of water resulted in a relatively high comprehensive score. Therefore, 6, 10, and 14 were selected as the levels for the water addition ratio in the orthogonal process investigation.

[0175] 2.2.5 Examination of extraction time

[0176] Four samples of gallnut, pomegranate peel, and pomegranate flower were taken and extracted three times with ten times the amount of water. The extraction times were set to 0.5, 1.0, 1.5, and 2.0 hours, respectively. After filtration, the filtrates were combined and the total volume of the filtrates was measured. The yield of extract, tannin, gallic acid, and ellagic acid content were determined and comprehensively scored. The results are shown in Table 5.

[0177] Table 5 Results of the Extraction Time Examination

[0178] Extraction time (h) Overall score 0.5 58.96 1.0 77.87 1.5 92.67 2.0 92.32

[0179] The results showed that the comprehensive extraction rates of the three medicinal materials were relatively high at 1.0, 1.5, and 2.0 hours, so they were selected as the extraction time levels for orthogonal process investigation.

[0180] 2.2.6 Examination of the number of extractions

[0181] Gallnut seeds, pomegranate peel, and pomegranate flowers were sampled according to the prescribed amounts and extracted according to the process design in Table 6. After filtration, the filtrates were combined and the total volume of the filtrates was measured separately. The yield, tannin content, gallic acid content, and ellagic acid content were determined using the same method as described above. The results are shown in Table 7.

[0182] Table 6. Experimental Arrangement for the Number of Extractions

[0183]

[0184]

[0185] Table 7 Results of the number of extractions

[0186] Number of extractions (times) Overall score 1 95.29 2 90.69 3 85.69 4 78.33

[0187] The results showed that extraction was basically complete after three extractions. Therefore, the number of extractions (1, 2, and 3) was selected as the level of extraction number for the orthogonal process investigation.

[0188] 2.3 Orthogonal Experiment

[0189] Based on the factor level range determined by the results of the single-factor experiment, L9(3) was selected. 4The orthogonal array and factor level table are shown in Table 8. Gallnut, pomegranate peel, and pomegranate flower were weighed at 1 / 10 of the prescription amount and extracted using the orthogonal scheme described below. After filtration, the filtrates were combined and diluted to an integer volume. The extract yield, tannin content, and gallic acid content were determined using the same method as described above. Orthogonal analysis was then performed after comprehensive scoring. Specific results are shown in Table 9.

[0190] Table 8 Factor Level Table

[0191]

[0192] Table 9. Comprehensive scoring results of orthogonal experiments

[0193]

[0194]

[0195] The intuitive analysis results show that the influence of each factor on the experimental results is A>C>B, and the optimal level for each factor is A3B3C3. Analysis of variance results show that factor A has a statistically significant impact on the experimental results (P<0.05), therefore A3 is selected; factors B and C have no statistically significant impact on the experimental results (P>0.1), and considering time and cost savings, B2C2 is selected. The optimal extraction process is A3B2C2, i.e., 10 times the amount of water added, 1.5 hours of extraction time, and 3 extractions.

[0196] 3. Research on impurity removal process

[0197] Commonly used methods include filtration, alcohol precipitation, and centrifugal precipitation. However, the medicinal materials in the prescription contain tannins that have varying degrees of inhibitory effects on viruses, fungi, and bacteria. Therefore, maintaining the original extraction process makes alcohol precipitation unsuitable. Plate and frame filtration involves numerous and cumbersome steps. Centrifugation is simple to operate, has good impurity removal effects, and minimizes the loss of active ingredients. Therefore, high-speed centrifugation was chosen to remove solid particulate impurities from the aqueous extract. High-speed centrifugation is a physical separation method that utilizes the centrifugal force generated by the high-speed rotation of a centrifuge to separate solids and liquids, or two immiscible liquids with different densities, by creating different centrifugal forces. The degree of separation is directly related to the centrifuge speed; the higher the speed, the more impurities are removed. To ensure efficacy and control the dosage of the preparation, the centrifugation speed and centrifugation time were investigated.

[0198] Considering that the water extraction process uses the content of gallic acid, ellagic acid, and tannins as evaluation indicators, and these are also large molecules, water extraction with alcohol precipitation or the addition of clarifying agents will remove these active ingredients. Therefore, this experiment only used four methods—natural sedimentation, overnight standing at 4℃, and centrifugation at different speeds and times—to study the impurity removal process of the water extract. The effects of different impurity removal processes on the content of gallic acid, ellagic acid, and tannins in the extract and the yield of the extract were investigated, and a reasonable and feasible impurity removal process was determined in conjunction with large-scale industrial production.

[0199] 3.1 Preparation of sample solution

[0200] Weigh out 31.5g of gallnut, 18.9g of pomegranate peel and 18.9g of pomegranate flower, mix them well, extract them using the optimal water extraction process, combine the extracts and mix them well for later use.

[0201] 3.2 Impurity Removal Methods

[0202] Take 200 ml of each of the mixed extracts and investigate the impurity removal methods according to the method in Table 10. Determine the content of gallic acid, ellagic acid, tannins and the yield of extract. The results are shown in Table 11.

[0203] Table 10 Impurity Removal Process Methods

[0204] Serial Number Sample Name Specific methods 1 Natural settling Place at room temperature (25℃) for 12 hours 2 Let stand at low temperature Let stand at 4℃ for 12 hours 3 High-speed centrifugation (1500 r / min) Centrifuge at 1500 rpm for 10 minutes 4 High-speed centrifugation (3000 r / min) Centrifuge at 3000 rpm for 10 minutes

[0205] Table 11 Results of paste yield and content after impurity removal process

[0206] Impurity removal methods Overall score Natural settling 97.98 Let stand at low temperature 108.01 Centrifuge at 1500 rpm 102.74 Centrifuge at 3000 rpm 98.7

[0207] The experimental results show that the low-temperature static setting method has the highest comprehensive score, but it takes the longest time and only has a significant impact on the paste yield. The centrifugation method at 1500 rpm has a similar filtration effect and is more time-saving and labor-saving. Therefore, centrifugation at 1500 rpm for 10 minutes is selected as the best impurity removal process.

[0208] 4. Research on Concentration and Drying Process

[0209] Atmospheric pressure water bath concentration is time-consuming, and the active ingredients are easily destroyed by prolonged heating; atmospheric pressure drying is also time-consuming and uneven. Therefore, to improve production efficiency and prevent the active ingredients from being destroyed by prolonged heating, vacuum concentration and drying equipment is used for the concentration and drying of water extracts.

[0210] 4.1 Concentration process of water extract

[0211] According to the prescription ratio, weigh 31.5g of gallnut, 18.9g of pomegranate peel, and 18.9g of pomegranate flower, respectively. Extract using the optimal water extraction process, filter (140 mesh), combine the filtrates, centrifuge at 1500 rpm for 10 minutes, and add water to a final volume of 2000ml. Accurately measure an appropriate amount and determine the contents of gallic acid, ellagic acid, and tannins. Then, measure three 500mL portions and concentrate them under reduced pressure at 50℃, 60℃, and 70℃ respectively to obtain a clear extract with a relative density of 1.15-1.20 (60℃), and bring the volume to the same level. Accurately measure an appropriate amount of each concentrate and determine the contents of gallic acid, ellagic acid, and tannins. The results are shown in Table 12.

[0212] Table 12 Results of Concentration Process Investigation

[0213] Concentration temperature (°C) Overall score 50 108.15 60 102.26 70 62.07

[0214] Based on the experimental results, the optimal concentration method for the water extract was determined to be vacuum concentration at 50℃.

[0215] 4.2 Drying process of water extract

[0216] From the concentrated extract obtained by the aforementioned optimal concentration process, take equal masses of clear extract (the reason for choosing mass rather than volume is that clear extract with a relative density of around 1.2 is relatively viscous; taking equal volumes would result in significant losses during transfer, making direct weighing more convenient). Dry these extracts under vacuum at 50, 60, and 70°C (-0.08 MPa) to obtain dry extracts for later use. Accurately weigh appropriate amounts from each of the three dry extracts and determine the contents of gallic acid, ellagic acid, and tannins. The results are shown in Table 13.

[0217] Table 13 Results of Drying Process Investigation

[0218] Drying temperature (°C) Ointment yield 50 89.97 60 92.72 70 99.09

[0219] Based on the experimental results, the optimal drying temperature for the water extract concentrate was selected as 70℃.

[0220] 5. Molding process research

[0221] After suppository administration, the drug must be melted in the matrix before it can be released from the matrix, dispersed at the site of action, and finally absorbed through contact with the mucous membrane. Therefore, the type and properties of the matrix directly affect the drug release rate.

[0222] Since this preparation is a vaginal suppository, according to the provisions of Appendix IW of the 2015 edition of the Chinese Pharmacopoeia regarding suppositories, vaginal suppositories are duckbill-shaped, spherical, or oval. Most commercially available vaginal suppositories are also duckbill-shaped. Therefore, the shape of this suppository is tentatively determined to be duckbill-shaped, and a water-soluble matrix is ​​selected as the main matrix to examine its formability.

[0223] 5.1 Determination of Molding Process Evaluation Indicators

[0224] The overall score is the sum of the scores of the above indicators, with appearance (Q1) and melting time (Q2) as evaluation indicators.

[0225] Q1: Place the suppository on a piece of white paper and observe its appearance under natural light. The observation criteria include the suppository's integrity, color uniformity, surface smoothness, and the comfort of touch. A score of 10 is given for a complete appearance, uniform color, smooth surface, no bubbles, and suitable hardness; 8 points are given for a few bubbles, a rough or dry surface, or slight surface defects; 5 points are given for a large number of bubbles, excessive hardness, or surface defects; and 0 points are given for uneven color, cracks, or easy crumbling.

[0226] Q2: The melting time test method is determined according to Appendix XIIB of the 2010 edition of the Chinese Pharmacopoeia, Part II. The Pharmacopoeia stipulates that all three suppositories with a fatty base should completely melt, soften, or have no hard core upon touch within 30 minutes. Therefore, for ease of scoring, 10 points are awarded for complete dissolution within 30 minutes; 9 points for most dissolution (≥50%); 8 points for partial dissolution (<50%); 7 points for softening but not dissolving; and 0 points for softening but with a hard core.

[0227] 5.2 Investigation of substrate types

[0228] PEG400, 4000, 1500, and 6000 were selected and mixed with S40 to form suppositories. The suppository formation effect of each PEG-S40 matrix ratio was investigated, and three of them were selected for further investigation.

[0229] As shown in Table 14, weigh out the extract powder, catechu and alum powder, mix them and set aside. Weigh out each PEG and mix them with S40 in the specified proportions. Heat in a 65°C water bath until melted. Under the condition of keeping warm in a 65°C water bath, add the above-mentioned powder mixture to each mixture. Stir and mix thoroughly. Pour into a mold coated with lubricant (3.0g), with the liquid level slightly overflowing the mold opening by 1-2mm. Refrigerate in a 5°C refrigerator for 10 minutes. Remove, flatten with a knife, open the mold, and remove.

[0230] Table 14 Experimental Arrangement for Matrix Type Investigation

[0231]

[0232] Table 15 Comprehensive Scoring Results of PEG400+S-40 Evaluation

[0233]

[0234] As shown in Table 15, as s-40 increases, the surface gloss decreases and the hardness increases.

[0235] Table 16: Comprehensive Scoring Results of PEG1500+S-40 Evaluation

[0236]

[0237] As shown in Table 16, the melting time of PEG1500 and S-40 is too long when used together, so it is not considered.

[0238] 5.3 Orthogonal Experimental Investigation

[0239] To ensure the accuracy and repeatability of the experiment, an orthogonal experimental design was designed (as shown in Table 17), and the matrix ratio was optimized.

[0240] Table 17 Factor Level Table

[0241]

[0242]

[0243] Table 18 Results of the Orthogonal Experiment

[0244]

[0245] As shown in Table 18, the comprehensive evaluation of factors such as melting time, appearance, hardness and ease of casting is conducted. The comprehensive ratio that affects the quality of plug formation is PEG400:PEG4000:PEG6000:S-40 = 3:1:2:2.

[0246] 5.4 Optimization of Basic Matrix Formulation Function

[0247] Given the hygroscopic nature of polyethylene glycol-based matrices, this experiment investigated the amount of glycerin added to reduce hygroscopicity and improve the suppositories' flexibility and wettability. The results were examined in terms of appearance and melting time. The following steps were taken: Extract powder, catechu, and alum powder were weighed and mixed. Five portions of S-40 were weighed separately and melted in a 60°C water bath. Under the same 60°C water bath holding condition, the above-mentioned powder mixture was added to each portion, and the mixture was thoroughly stirred and mixed. The mixture was then poured into a suppository mold coated with lubricant (3.0g), with the liquid level slightly overflowing the mold opening by 1-2mm. The suppositories were then refrigerated at 5°C for 10 minutes. Afterward, the liquid was removed, leveled with a knife, and the mold was opened. The appearance and melting time of the prepared suppositories were observed, and a comprehensive score was determined. The results are shown in Table 19.

[0248] Table 19 Experimental Arrangement for Investigating Glycerin Dosage

[0249]

[0250]

[0251] As shown in Table 19, the melting time of the suppositories gradually decreased with the increase of glycerol content. When the ratio of matrix composition to glycerol was from 10:1 to 8:3, the suppositories produced had advantages such as round and smooth shape, uniform color, and suitable hardness. However, when the ratio was 7:4, the suppositories became softer and slightly sticky.

[0252] 5.5 Molding process screening

[0253] Suppository bases containing polyethylene glycol are greatly affected by temperature and stirring time during preparation. Therefore, the preparation temperature and stirring time were the main factors considered in the suppository molding process.

[0254] 5.5.1 Effect of Melting Temperature on Suppository Formation

[0255] The matrix melting was investigated at temperatures of 60℃, 65℃, 70℃, and 75℃. After the matrix melted, it was mixed with extract powder to form suppositories under a certain stirring time. The melting time, the degree of uniformity between the drug and the matrix, the ease of filling the mold, the cooling time, and the appearance were used as evaluation indicators. The results are shown in Table 20.

[0256] Table 20 Effect of preparation temperature on suppository formation

[0257]

[0258] Mixing degree: ++++ Very easy to mix, +++ Somewhat easy to mix, ++ Easy to mix, - Difficult to mix, matrix stratification, drug spherical formation.

[0259] Therefore, 60℃-65℃ is the optimal temperature for dissolving the matrix.

[0260] 5.5.2 Effect of stirring time on suppository formation

[0261] After the matrix was melted at 60-65℃, the appearance characteristics were examined after the stirring time following the addition of the drug powder. The results are shown in Table 21.

[0262] Table 21 Effect of stirring time on suppository formation

[0263]

[0264] The results show that, under certain rotation speed and preparation temperature conditions, the longer the stirring time, the better the uniformity of the mixture between the drug and the matrix, and the better the flowability and the absence of air bubbles. However, if the stirring time is too short, the drug and the matrix will not be fully mixed. Therefore, considering energy consumption and production cycle, in actual production, the stirring time should be determined to be until the drug powder and the matrix are fully mixed and free of particles and lumps.

[0265] 6. Process Confirmation

[0266] Weigh the medicinal materials according to the following proportions: catechu, alum, gallnut, pomegranate peel, and pomegranate flower in a ratio of approximately 3.15:1.26:3.15:1.89:1.89. Mix the catechu and alum and grind them into a fine powder for later use. Grind the gallnut, pomegranate peel, and pomegranate flower into a coarse powder, add 10 times the amount of water, and extract three times, 1.5 hours each time. Combine the extracts and centrifuge at 1500 rpm for 10 minutes. Concentrate the supernatant under reduced pressure (0.08 MPa, 50℃) until the relative density is between 1.15 and 1.20 (6). Vacuum dried (0.08MPa, 70℃) between 0℃ and 0℃, pulverized into a fine powder, and mixed with the above-mentioned fine powder of medicinal materials. Each matrix component was weighed according to the following ratio: PEG400:PEG4000:PEG6000:S-40 (3:1:2:2):glycerin (9:2). The mixture was melted in a 60℃ water bath, and then slowly added to the mixed medicinal powder in a constant temperature water bath at 60-65℃. The mixture was stirred for at least 20 minutes to ensure uniform mixing. The mixture was then poured into 3.0g duckbill-shaped suppositories, cooled, and made into 1000 suppositories. The suppositories were then packaged. The flowchart is shown below. Figure 1 As shown.

[0267] Pharmacological Examples

[0268] 1. Experimental Materials

[0269] 1.1 Experimental reagents

[0270] 1.1.1 Test Drug

[0271] The preparation process of pomegranate flower sap is derived from the above-mentioned "6 process determinations".

[0272] High-dose sample of pomegranate flower suppositories for rat experiments, brownish-brown suppositories. Each suppository weighs 0.18g, and each gram is equivalent to 0.378g of raw medicinal material. Batch number: 170960. Storage: Store in a sealed container at room temperature. Provided by: Xinjiang Uygur Autonomous Region Uygur Medicine Research Institute.

[0273] Low-dose sample of pomegranate flower suppositories for rat experiments: light brown suppositories. Each suppository weighs 0.18g, and each gram is equivalent to 0.133g of raw medicinal material. Batch number: D1022. Storage: Store in a sealed container at room temperature. Provided by: Xinjiang Uygur Autonomous Region Uygur Medicine Research Institute.

[0274] 1.1.2 Matrix control

[0275] Pomegranate flower suppository excipient sample: white suppository. Weight per suppository: 0.17g. Composition: poly(40) stearate, polyethylene glycol 400, polyethylene glycol 4000, polyethylene glycol 6000, glycerin. Batch number: K1023. Storage: Store in a sealed container at room temperature. Provided by: Xinjiang Uygur Autonomous Region Uygur Medicine Research Institute.

[0276] 1.1.3 Positive control drug

[0277] The prescription for Xiaomi Suppositories includes: ginseng stem and leaf saponins, Lithospermum erythrorhizon, Phellodendron chinense, Sophora flavescens, alum, borneol, and catechu. Appearance: This product is a brown to dark brown suppository with a characteristic odor. Indications: Clears heat and detoxifies, dries dampness and kills parasites, removes necrotic tissue and promotes tissue regeneration. Used for leukorrhea caused by damp-heat accumulation, characterized by profuse, yellow, thick, and foul-smelling discharge, and vulvar itching; also for trichomonal vaginitis, fungal vaginitis, nonspecific vaginitis, and cervical erosion with the above symptoms. Specifications: 3g per suppository. Dosage and Administration: Vaginal administration, one suppository once daily. Storage: Store in a tightly closed container below 30℃. Approval Number: National Medicine Approval Number Z22021281. Batch Number: 190509. Manufacturer: Tongyao Pharmaceutical Group Co., Ltd.

[0278] Sample of Xiaomi Suppository for Rat Experiment: Brownish-brown suppository. Each suppository weighs 0.18g, and each gram of suppository is equivalent to 0.875g of raw medicinal material. Storage: Store in a sealed container at room temperature. Provided by: Xinjiang Uygur Autonomous Region Uygur Medicine Research Institute.

[0279] In the experiment, 0.06g was used as the low dose, which is the clinically equivalent dose for rats; and 0.12g was used as the high dose, which is twice the clinically equivalent dose for rats.

[0280] 1.1.4 Dosage setting

[0281] Dosage setting basis: Based on the clinically intended dosage provided by the client, the equivalent dosage for different animal species was calculated according to body surface area, and the clinical equivalent dosage was set as the low dose. The adult vaginal administration dosage of pomegranate flower suppositories is 3g / day, each gram is equivalent to 0.378g of raw medicinal material, i.e., 1.134g of raw medicinal material / person·day. According to the animal-to-human dosage conversion table, the rat dosage is calculated to be 0.12g / kg·d. The rat experimental samples provided by the client are: ① Pomegranate flower suppositories: each suppository is equivalent to 0.068g of raw medicinal material, which is 2.8 times the clinical equivalent dosage for rats, and is set as the high dose; ② Low concentration of pomegranate flower suppositories: each suppository is equivalent to 0.024g of raw medicinal material, which is the clinical equivalent dosage for rats, and is set as the low dose. The mouse dosage was converted from the clinical equivalent dosage as the low dose, and 2.8 times that was taken as the high dose. The clinical dose of the positive-positive drug Xiaomi Suppository is 3g / suppository. Each gram is equivalent to 0.875g of raw medicinal materials, i.e., 2.625g of raw medicinal materials / person / day. The clinically equivalent dose is taken as the low dose, and twice that is taken as the high dose. The dosage settings are shown in Table 22.

[0282] Table 22 Dosage Settings

[0283]

[0284] In mouse experiments, the rat experimental samples were melted in a 50°C water bath and then administered topically at the mouse equivalent dose.

[0285] 1.2 Laboratory Animals

[0286] 1.2.1 Strains and Sources of Laboratory Animals

[0287] Kunming mice, female, weighing 18 - 22 g; Wistar rats, female, weighing 180 - 220 g, provided by the Animal Experiment Center of Xinjiang Medical University, with the production license number of experimental animals being SCXK(Xin)2018 - 0002.

[0288] 1.2.2 Rearing of Laboratory Animals

[0289] After being caged separately, the laboratory animals were reared conventionally with free access to food and water.

[0290] Feed: Co60 - irradiated maintenance feed for experimental mice, produced by Jiangsu Medison Biomedical Co., Ltd., with the production license number: Su Feed License (2018)10030.

[0291] Drinking water: Special water for laboratory animals.

[0292] 1.2.3 Rearing Environment Temperature of Laboratory Animals: 20 - 26°C

[0293] Relative Humidity: 40% - 60%

[0294] Lighting Conditions: 12h / 12h light - dark alternation Ventilation Condition: Fresh air

[0295] General Cleaning and Disinfection of the Animal House: Mopping and cleaning with 0.2% bromogeramine.

[0296] 1.3 Reagents and Others [

[0297] Injection of Estradiol Benzoate, produced by Sichuan Jinke Pharmaceutical Co., Ltd., batch number: 20200301; Injection of Hydrocortisone, produced by Tianjin KingYork Pharmaceutical Co., Ltd., batch number: 1910241; Injection of Gentamicin Sulfate, produced by Yichang Humanwell Pharmaceutical Co., Ltd., batch number: 91Y05031; Injection of Sodium Chloride, produced by Xinjiang Pharmaceutical Co., Ltd., China National Pharmaceutical Corporation, batch number: 1912047; Xylene, produced by Tianjin Yongcheng Fine Chemical Co., Ltd., batch number: 20190508; Glacial Acetic Acid, produced by Tianjin Fuchen Chemical Reagent Factory, batch number: 2013031; Evans Blue, sub - packed by Shanghai Chemical Reagent Procurement and Supply Station, batch number: 20120917; Compound Dextran 40 Injection, produced by Xi'an Wanlong Pharmaceutical Co., Ltd., batch number: 8190415. Candida albicans, number: CMCC(F)98001, from the National Institutes for Food and Drug Control, batch number: 98001 - 2a20.

[0298] 1.4 Instruments

[0299] BS200S electronic balance, instrument number: 11536806, Beijing Sartorius Balance Co., Ltd.; YP6000N electronic balance, instrument number: C011304056, Shanghai Jinghai Instrument Co., Ltd.; AL 204 electronic balance, instrument number: 1229520103, Mettler Toledo Instruments (Shanghai) Co., Ltd.; SP-752 UV-Vis spectrophotometer, instrument number: ZWO 308102309, Shanghai Spectrometer Instrument Co., Ltd.; DSX-30L portable autoclave, instrument number: 180076, Shanghai Shenan Medical Instrument Factory; Auto MS1000 mass spectrometer, Zhengzhou Antu Bioengineering Co., Ltd.

[0300] 2 Experimental Methods and Results

[0301] 2.1 Antibacterial effect - Effect on fungal vaginitis in rats

[0302] 2.1.1 Establishment of a rat model of fungal vaginitis

[0303] Eighty female Wistar rats, weighing 180–220 g, were used. After 10 days of acclimatization, all but one of the ten rats in the normal control group underwent modeling. From day 1 of modeling, each rat was subcutaneously injected with 0.1 mL of 2 mg / mL estradiol benzoate injection once daily for four consecutive days, followed by injections every other day until the modeling process ended. From day 3 of modeling, each rat was subcutaneously injected with 0.5 mL of 5 mg / mL hydrocortisone injection every other day for three days. On day 5 of modeling, the rats' vaginas were first irrigated twice with 0.25 mL of 80,000 IU gentamicin sulfate injection, followed by the injection of 0.15 mL of Candida albicans solution. The rats were then inverted for approximately 3 minutes to prevent leakage of the solution. This solution was injected into the vagina once daily for five consecutive days. Four days after the bacterial infection, vaginal secretions were collected with a sterile swab and smeared on the surface of Sabouraud dextrose agar. The agar was incubated at 28°C for 48 hours. White colonies of varying sizes were visible to the naked eye and identified as Candida albicans by an Auto MS1000 mass spectrometer, indicating that the model was successfully established.

[0304] 2.1.2 Grouping and Administration

[0305] Except for a normal control group of 10 rats, 60 rats with successfully established pomegranate flower suppositories were randomly divided into 5 groups: model group, high-dose pomegranate flower suppository group, low-dose pomegranate flower suppository group, high-dose pomegranate flower suppository group, and low-dose pomegranate flower suppository group, with 12 rats in each group. Rats in each treatment group were administered the medication vaginally, while the normal control group and model group were given the same amount of pomegranate flower suppository excipient, once a day for 5 consecutive days.

[0306] 2.1.3 Detection Indicators

[0307] Twenty-four hours after the last administration, vaginal secretions were collected using a sterile swab and smeared onto the surface of Sabouraud dextrose agar. The agar was incubated at 28°C for 48 hours, and colony formation was observed and counted visually. Evaluation criteria were: 0: no colonies; 1+: 1–10 colonies; 2+: 11–100 colonies; 3+: more than 100 colonies. Results are shown in Table 23.

[0308] Table 23. Count of Candida albicans colonies in the vagina of rats in each group.

[0309] Cage number serial number Normal control Model Low dose of anti-inflammatory suppositories High dose of anti-inflammatory suppositories Low dose of flower sap High dose of flower stalk ♀1 1 0 1+ 0 0 0 0 2 0 0 0 0 1+ 0 3 0 0 0 1+ 0 0 4 0 0 2+ 0 0 0 5 0 0 0 3+ 2+ 0 6 － 2+ 0 0 0 0 ♀2 1 0 2+ 0 0 0 0 2 0 3+ 0 0 0 0 3 0 1+ 3+ 1+ 0 1+ 4 0 0 0 0 0 1+ 5 0 1+ 0 0 0 0 6 － 0 0 0 0 0

[0310] The infection rate and inhibition rate of Candida albicans vaginitis in each group of rats were calculated according to Table 23, and the statistical results are shown in Table 24.

[0311] Table 24 Effects of pomegranate flower suppositories on a model of fungal vaginitis

[0312]

[0313] Note: Compared with the normal control group, **P<0.01.

[0314] As shown in Table 24, compared with the normal control group, the vaginal Candida albicans infection rate of rats in the model group was significantly increased (P<0.01); compared with the model group, both high and low doses of pomegranate flower suppositories could reduce the vaginal Candida albicans infection rate in rats, indicating that it has a certain therapeutic effect on Candida albicans-induced vulvovaginitis in rats.

[0315] 2.2 Anti-inflammatory experiment - Effect of xylene on mouse ear swelling

[0316] Fifty female Kunming mice, weighing 18–22 g, were randomly divided into five groups (n=10 each) after 3 days of acclimatization feeding: a model group, a low-dose Xiaomishuan group, a high-dose Xiaomishuan group, a low-dose Shiliuhuashuan group, and a high-dose Shiliuhuashuan group. The drugs were applied to the right ear of each group, while the model group received an equal amount of Shiliuhuashuan excipient. Administered twice daily, 1 hour apart. One hour after the last administration, the drugs were wiped off with warm water, and 20 μL of xylene was evenly applied to both sides of the right auricle of each group to induce inflammation. The left ear served as a control. After 30 minutes, the mice were euthanized by cervical dislocation, and circular ear pieces were punched symmetrically from both ears using a 6 mm diameter punch, and accurately weighed. The results are shown in Table 25.

[0317] Table 25. Weights (mg) of left and right ear pieces of mice in each group.

[0318]

[0319]

[0320] Note: Data from model group ♀1 cage 4 and high-dose Xiaomishuan group ♀1 cage 1 showed high dispersion, so no statistics were performed.

[0321] The ear swelling degree and swelling inhibition rate of mice in each group were calculated according to Table 25, and the statistical results are shown in Table 26.

[0322] Swelling degree = Weight of right earpiece - Weight of left earpiece. Swelling inhibition rate (%) = (Mean swelling degree of model group - Mean swelling degree of treatment group) / Mean swelling degree of model group × 100%

[0323] Table 26 Effects of pomegranate flower suppositories on xylene-induced ear swelling in mice (x±S)

[0324]

[0325] Note: Compared with the model group, *P<0.05, **P<0.01.

[0326] As shown in Table 26, compared with the model group, both high and low doses of pomegranate flower suppository reduced xylene-induced ear swelling in mice, indicating that it could alleviate inflammatory swelling of the mouse auricle. The low dose group of pomegranate flower suppository showed a significant effect (P<0.05).

[0327] 2.3 Anti-inflammatory experiment - Effect on acetic acid-induced increase in peritoneal capillary permeability in mice [6-8]

[0328] Sixty female Kunming mice, weighing 18–22 g, were used and acclimatized for 3 days. They were then randomly divided into 5 groups of 12 mice each, as described in section 2.2. The day before the experiment, the neck and back hair of each group was shaved to approximately 2 cm × 2 cm. On the day of the experiment, the drug was applied to the shaved skin of each treatment group, while the model group received an equal amount of pomegranate flower suppository excipient. The drugs were administered twice daily, with a 1-hour interval between administrations. One hour after the last administration, the drugs were wiped off with warm water. Each mouse was then injected intravenously with 10 mL / kg of 0.5% Evans blue saline solution, followed by an intraperitoneal injection of 0.2 mL / mouse of 0.8% glacial acetic acid saline solution. Twenty minutes later, the mice were euthanized by cervical dislocation. The peritoneal cavity was repeatedly aspirated and flushed with 6 mL of saline solution. The flushing fluid was aspirated, centrifuged at 3000 rpm for 10 minutes, and the supernatant was measured at 590 nm. The results are shown in Table 27.

[0329] Table 27 Absorbance values ​​(A) of peritoneal fluid from each group of mice

[0330]

[0331] Note: Females in cage 1, size 6 of the low-dose Xiaomishuan group and females in cage 2, size 5 of the low-dose Shiliuhuashuan group had intra-abdominal bleeding, and their absorbance values ​​were not measured; the data of females in cage 2, size 5 of the model group, females in cage 2, size 3 of the low-dose Xiaomishuan group, females in cage 1, size 5 of the low-dose Shiliuhuashuan group, and females in cage 2, size 6 of the high-dose Shiliuhuashuan group showed large dispersion, so the above data were not statistically analyzed.

[0332] The inhibitory rate of the drug on the increased permeability of peritoneal capillaries in mice was calculated according to Table 27, and the statistical results are shown in Table 28.

[0333] Inhibition rate (%) = (Mean absorbance of model group - Mean absorbance of drug-treated group) / Mean absorbance of model group × 100%

[0334] Table 28 Effects of pomegranate flower suppositories on acetic acid-induced increase in peritoneal capillary permeability in mice (x±S)

[0335]

[0336] As shown in Table 28, compared with the model group, both high and low doses of pomegranate flower suppositories significantly reduced the absorbance value of peritoneal fluid in mice (P<0.05 or P<0.01), indicating that they inhibited the increase in peritoneal capillary permeability induced by acetic acid in mice, suggesting that they have a good anti-inflammatory exudative effect.

[0337] 2.4 Antipruritic Experiment - Effect on Dextran-Induced Skin Itching in Mice

[0338] Fifty female Kunming mice, weighing 18–22 g, were used and grouped as described in section 2.2. The day before the experiment, the neck and back hair of each group of mice was shaved to approximately 2 cm × 2 cm. On the day of the experiment, the drug was applied to the shaved skin of each treatment group, while the model group received an equal amount of pomegranate flower suppository excipient. The drugs were administered twice daily, with a 1-hour interval between administrations. One hour after the last administration, the drugs were wiped off with warm water. Each mouse was injected intravenously with dextran at 1.25 mg / kg via the tail vein. Itching was indicated by the mice scratching their heads with their forepaws, scratching their trunks with their hind paws, and biting various parts of their bodies. The number of scratches and the duration of itching within 30 minutes were observed and recorded. The results are shown in Tables 29 and 30.

[0339] Table 29 Number of times mice in each group experienced itching

[0340]

[0341]

[0342] Table 30 Total duration of itching in mice of each group (s)

[0343]

[0344] The statistical results of the above data are shown in Table 31.

[0345] Table 31 Effects of pomegranate flower suppositories on dextran-induced pruritus in mice (x±S)

[0346]

[0347] Note: Compared with the model group, *P<0.05.

[0348] As shown in Table 31, compared with the model group, both high and low doses of pomegranate flower suppositories reduced the frequency and total duration of skin itching in mice, indicating that they have an inhibitory effect on dextran-induced skin itching in mice. The high-dose group of pomegranate flower suppositories showed a significant effect (P<0.05), suggesting a good antipruritic effect.

[0349] 2.5 Statistical Methods

[0350] SPSS 17.0 statistical software was used to perform statistical analysis on the data of each group. Quantitative data were expressed as mean ± standard deviation (x ± S). The chi-square test was used to compare rates between groups. One-way ANOVA was used for the remaining groups. P < 0.05 was considered statistically significant.

[0351] 3. Conclusion

[0352] The above experimental results indicate that pomegranate flower suppositories can reduce the Candida albicans infection rate in rats with fungal vaginitis, alleviate xylene-induced ear swelling in mice, and inhibit acetic acid-induced increased peritoneal capillary permeability in mice. Furthermore, pomegranate flower suppositories can reduce the frequency and duration of dextran-induced skin itching in mice. These results suggest that pomegranate flower suppositories have good inhibitory, anti-inflammatory, and antipruritic effects against Candida albicans.

[0353] The embodiments of the present invention have been described in detail above. Specific examples have been used to illustrate the principles and implementation methods of the present invention. The descriptions of the embodiments above are only for the purpose of helping to understand the method and core ideas of the present invention. Furthermore, any changes or modifications made by those skilled in the art based on the ideas of the present invention, its specific implementation methods, and its application scope, are all within the scope of protection of the present invention. Therefore, the content of this specification should not be construed as a limitation of the present invention.

Claims

1. A suppository containing pomegranate flowers, characterized in that, The suppository is made of catechu, alum, gallnut, pomegranate peel, pomegranate flower, a first excipient and a second excipient, wherein the first excipient is polyethylene glycol and polyoxyethylene (40) monostearate, and the second excipient is glycerin; Wherein, the polyethylene glycol is polyethylene glycol 400, polyethylene glycol 4000 and polyethylene glycol 6000; The mass ratio of polyethylene glycol 400, polyethylene glycol 4000, polyethylene glycol 6000 and polyoxyethylene (40) monostearate is 3:1:1:

2. The mass ratio of the catechu, the alum, the gallnut, the pomegranate peel, the pomegranate flower, the first excipient, and the second excipient is (2~4):(0.5~2):(2~4):(1~3):(1~3):(20~30):(2~8).

2. The suppository according to claim 1, characterized in that, The mass ratio of the catechu, the alum, the gallnut, the pomegranate peel, the pomegranate flower, the first excipient, and the second excipient is (2.9925~3.3075):(1.197~1.323):(2.9925~3.3075):(1.7955~1.9845):(1.7955~1.9845):(23.845~26.355):(5.301~5.859).

3. A method for preparing the suppository according to claim 1 or 2, characterized in that, The method includes the following steps: (1) Weigh out an appropriate amount of catechu and alum, mix them and grind them into fine powder, and optionally sieve them to obtain the first mixture; (2) Weigh out appropriate amounts of gallnut, pomegranate peel and pomegranate flower, mix them and crush them into coarse powder, add solvent for extraction, centrifuge to take the supernatant, concentrate, dry, crush into fine powder, and optionally sieve to obtain the second mixture; (3) Mix the first mixture with the second mixture to obtain a third mixture; (4) Weigh appropriate amounts of the first and second excipients, melt them in a water bath, stir and add them to the third mixture, mix evenly to obtain the fourth mixture; and (5) The fourth mixture is injected into a suppository mold, cooled, and granulated to obtain the suppository.

4. The method according to claim 3, characterized in that, In step (2), the solvent is water or ethanol.

5. The method according to claim 4, characterized in that, The solvent is water.

6. The method according to claim 3, characterized in that, The volume of the solvent is 6 to 14 times the weight of the coarse powder.

7. The method according to claim 6, characterized in that, The volume of the solvent is 9.5 to 10.5 times the weight of the coarse powder.

8. The method according to claim 3, characterized in that, The extraction method is heating and reflux extraction.

9. The method according to claim 8, characterized in that, The temperature for the heating and reflux extraction is 50℃~100℃.

10. The method according to claim 9, characterized in that, The temperature for the heating and reflux extraction is 100°C.

11. The method according to claim 3, characterized in that, The extraction is performed 1 to 5 times.

12. The method according to claim 11, characterized in that, The extraction was performed 3 times.

13. The method according to claim 3, characterized in that, The extraction time is 1 hour / time to 2 hours / time.

14. The method according to claim 13, characterized in that, The extraction time is 1.425 hours / time to 1.575 hours / time.

15. The method according to claim 3, characterized in that, The centrifuge speed is 1000 r / min to 3000 r / min.

16. The method according to claim 15, characterized in that, The centrifuge speed is 1425 r / min to 1575 r / min.

17. The method according to claim 3, characterized in that, The centrifugation time is 8 min to 12 min.

18. The method according to claim 17, characterized in that, The centrifugation time is 9.5 min to 10.5 min.

19. The method according to claim 3, characterized in that, The concentration is a vacuum concentration.

20. The method according to claim 19, characterized in that, The pressure for vacuum concentration is 0.06 MPa to 1.00 MPa.

21. The method according to claim 20, characterized in that, The pressure for vacuum concentration is 0.076 MPa to 0.084 MPa.

22. The method according to claim 19, characterized in that, The temperature for vacuum concentration is 45℃~75℃.

23. The method according to claim 22, characterized in that, The temperature for vacuum concentration is 47.5℃~52.5℃.

24. The method according to claim 3, characterized in that, The supernatant was concentrated under reduced pressure to a relative density of 1.10–1.30 at 60°C.

25. The method according to claim 24, characterized in that, The supernatant was concentrated under reduced pressure to a relative density of 1.15–1.20 at 60°C.

26. The method according to claim 3, characterized in that, The drying process is vacuum drying.

27. The method according to claim 3, characterized in that, The drying pressure is 0.06 MPa to 1.00 MPa.

28. The method according to claim 27, characterized in that, The drying pressure is 0.076 MPa to 0.084 MPa.

29. The method according to claim 3, characterized in that, The drying temperature is 55℃~75℃.

30. The method according to claim 29, characterized in that, The drying temperature is 66.5℃~73.5℃.

31. The method according to claim 3, characterized in that, In steps (1) and (2), the sieve is a 100-mesh sieve.

32. The method according to claim 3, characterized in that, The melting water bath temperature is 60℃~70℃.

33. The method according to claim 3, characterized in that, The water bath temperature for stirring is 60℃~65℃.

34. The method according to claim 3, characterized in that, The stirring time is at least 20 minutes.

35. The method according to claim 3, characterized in that, In step (5), the mass of the suppository is 2.85~3.15 g.

36. The method according to claim 3, characterized in that, The suppository has a duckbill-shaped plug.

37. Use of the suppository according to claim 1 or 2 in the preparation of a medicament for the prevention and / or treatment of nonspecific vaginitis.

38. The use according to claim 37, characterized in that, The nonspecific vaginitis mentioned is fungal vaginitis.

39. Use of the suppository according to claim 1 or 2 in the preparation of a medicament for relieving inflammatory swelling.

40. Use of the suppository according to claim 1 or 2 in the preparation of a medicament for inhibiting inflammatory exudation.

41. Use of the suppository according to claim 1 or 2 in the preparation of a medicament for the prevention and / or treatment of pruritus.

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