Preparation method of pharmaceutical composition for treating adenomyosis

Abstract

The invention provides a preparation method of a pharmaceutical composition for treating adenomyosis. The preparation method comprises the following steps: a, weighing the following raw material medicines in parts by weight: 25-35 parts of sculellaria barbata, 25-35 parts of oldenlandia diffusa, 5-15 parts of spina gleditsiae, 10-20 parts of curcuma zedoary, 5-15 parts of ground beeltle, 10-20 parts of rhizoma curculiginis, 10-20 parts of epimedium and 15-25 parts of polyporus umbellatus; b, extracting spina gleditsiae, curcuma zedoary, oldenlandia diffusa and sculellaria barbata by C, extracting rhizoma curculiginis, herba epimedii, ground beetles and polyporus umbellatus with water; and d, combining the extracts in the steps b and c, concentrating, and adding pharmaceutically acceptable auxiliary materials or auxiliary components to prepare a pharmaceutically common preparation. Pharmacodynamic studies prove that the pharmaceutical composition can obviously improve uterine injury of experimental adenomyosis rats, and the pharmaceutical effect of the pharmaceutical composition is better than that of a traditional Chinese medicine clinical conventional water decoction preparation under the condition of equal dosage or 1 / 2 dosage, so that the pharmaceutical composition prepared by the innovative preparation method has better pharmaceutical effect than that of clinical water decoction, and the pharmaceutical composition is suitable for clinical application. The compound has remarkable application potential in preparation of drugs or hospital preparations.

Description

Technical Field

[0001] The present invention relates to a preparation method of a pharmaceutical composition for treating adenomyosis, belonging to the field of traditional Chinese medicine. Background Art

[0002] Adenomyosis (AM) is a common and difficult gynecological disease, primarily characterized by severe dysmenorrhea, abnormal uterine bleeding, and infertility. The incidence ranges from 5-70%. Choosing a treatment plan is a clinical challenge, especially when combined with infertility. Women with AM have a 43% lower clinical pregnancy rate and a threefold increased risk of miscarriage.

[0003] The etiology and pathogenesis of AM are still related to factors such as destruction of the uterine junctional zone, epithelial-mesenchymal transition, proliferation, regeneration and differentiation of stem cells induced by tissue damage stimulation, destruction of the local microenvironment causing immune response leading to local vasoconstriction and spasm and even local ischemia and inflammatory response leading to local hyperplasia and adhesion.

[0004] Treatment for amenorrhea (AM) primarily involves medication and surgery. Six common medications are: 1) Oral contraceptives inhibit gonadotropin production, causing endometrial atrophy. They are suitable for patients with mild AM who experience dysmenorrhea and are not seeking to conceive. However, long-term use increases the risk of thrombosis. 2) Commonly used androgen derivatives include danazol and gestrinone, which have some effect on relieving dysmenorrhea and menstrual irregularities. However, these symptoms are prone to recurrence after discontinuation, and the incidence of androgen adverse reactions is high, as well as liver function. 3) Mifepristone has been shown to be effective in treating AM based on its immunological mechanism, but in my country, it is only approved for the treatment of uterine fibroids. 4) The levonorgestrel-releasing intrauterine system can relieve AM-related dysmenorrhea, reduce menstrual flow, and correct secondary anemia. A common side effect is irregular vaginal bleeding, and it has no effect on lesion reduction. 5) GnRH-a: This suppresses pituitary gonadotropin secretion, reducing ovarian gonadal hormone secretion. Its more significant side effects include hot flashes, sweating, vaginal dryness, bone loss, decreased libido, and insomnia. 6) Synthetic progestins: By reducing the release of pituitary gonadotropins and acting directly on the lesions, they induce decidualization of the ectopic endometrium, leading to atrophy and reduction. However, these progestins have certain side effects, such as epigastric discomfort and weight gain. Currently, dydrogesterone is used in patients with amenorrhea and infertility, improving overall dysmenorrhea scores and promoting fertility.

[0005] Overall, these modern medical treatments can control dysmenorrhea, heavy menstrual flow, and temporarily suppress menstrual cycles, but they often fail to provide satisfactory results for women seeking pregnancy and may lead to recurrence. For perimenopausal or postmenopausal women who do not desire childbearing, surgical treatments are often used, which can cause functional trauma to the body. Consequently, many patients seek the proven efficacy of Traditional Chinese Medicine (TCM).

[0006] AM falls under the Traditional Chinese Medicine (TCM) categories of "dysmenorrhea," "lumps and masses," "infertility," and "menstrual disorders." Its causes can be categorized as pathogenic cold, qi reversal, qi deficiency, and qi stagnation. The pathogenesis is damage to the Chong and Ren meridians and abnormalities in the uterus's storage and drainage functions, which cause menstrual blood to overflow and become dislocated blood. Dislocated blood accumulates locally, eventually forming blood stasis. Blood stasis leads to pain, resulting in the symptoms and signs of progressive dysmenorrhea. Treatment options include oral administration of Chinese herbal medicine, enema, and acupuncture, primarily focusing on promoting blood circulation and removing blood stasis. Unlike Western medications, TCM treatments generally do not suppress the normal function of the gonadal axis and can improve the state of the endometrium and inhibit ectopic endometrial lesions.

[0007] Lu Fengyang explored the clinical application of Wang Chengrong's idea of fire and blood stasis based on Bailian Sanjie Decoction. The 2018 master's thesis of Chengdu University of Traditional Chinese Medicine disclosed that Bailian Sanjie Decoction has the effects of clearing heat and detoxifying, resolving blood stasis and dispersing nodules, and is used to treat gynecological diseases of fire and blood stasis type. The clinical medication method is decoction. The study found that the decoction cannot reflect the efficacy of the prescription. Summary of the Invention

[0008] In order to solve the efficacy problem of the current Bailian Sanjie Decoction, the present invention provides a new preparation method of Bailian Sanjie Decoction.

[0009] The present invention provides a method for preparing a pharmaceutical composition for treating adenomyosis, which comprises the following steps:

[0010] a. Weigh the API in the appropriate weight ratio:

[0011] 25-35 parts of Scutellaria barbata, 25-35 parts of Hedyotis diffusa, 5-15 parts of Spina Gleditsiae, 10-20 parts of Curcuma zedoaria, 5-15 parts of Eupolyphaga sinensis, 10-20 parts of Curculigo orchioides, 10-20 parts of Epimedium, and 15-25 parts of Polyporus;

[0012] b. Extraction of Gleditsia sinensis, Curcuma zedoaria, Hedyotis diffusa and Scutellaria barbata with organic solvents;

[0013] c. Curculigo, Epimedium, Eupolyphaga sinensis, and Polyporus umbellatus are extracted with water;

[0014] d. Combine the extracts from step b and step c, concentrate them, add pharmaceutically acceptable excipients or auxiliary ingredients, and prepare a commonly used pharmaceutical preparation.

[0015] Further preferably, the weight ratio of the raw material drug described in step a is:

[0016] 28-32 parts of Scutellaria barbata, 28-32 parts of Hedyotis diffusa, 8-12 parts of Spina Gleditsiae, 12-18 parts of Curcuma zedoaria, 8-12 parts of Eupolyphaga sinensis, 10-20 parts of Curculigo orchioides, 12-18 parts of Epimedium, and 18-22 parts of Polyporus.

[0017] More preferably, the weight ratio of the raw material drug described in step a is:

[0018] 30 parts of Scutellaria barbata, 30 parts of Herba Hedyotis diffusae, 10 parts of Spina Gleditsiae, 15 parts of Rhizoma Curcumae, 10 parts of Eupolyphaga Seu Steleophora, 15 parts of Curculigo orchioides, 15 parts of Herba Epimedii, 20 parts of Polyporus.

[0019] Wherein, the organic solvent in step b is ethanol, methanol, acetone, petroleum ether, benzene, chloroform, ether, ethyl acetate, or dichloroethane; preferably, the organic solvent is ethanol;

[0020] The extraction method described in step b is first leaching, then reflux extraction and ultrasonic extraction.

[0021] Further preferably, the ethanol is 40-90% ethanol, preferably, the ethanol is 50% ethanol; the material-liquid ratio of the medicinal material to the ethanol is 1 g:8-12 mL.

[0022] Wherein, the extraction method described in step c is first leaching, then reflux extraction or ultrasonic extraction;

[0023] The material-liquid ratio of medicinal materials to water is: 1g:6-10mL.

[0024] The soaking time is 1-2 hours;

[0025] The reflux extraction is performed 2-3 times, each time for 0.5-2 hours.

[0026] Wherein, the concentration method described in step d is reduced pressure concentration, and the concentration temperature is 60°C.

[0027] The preparations are tablets, capsules, granules, aqueous solutions, tinctures, micropills, microspheres, dripping pills, controlled-release preparations, and sustained-release preparations.

[0028] The beneficial effects of the present invention are:

[0029] The pharmaceutical composition provided by the present invention has been shown in pharmacodynamic studies to significantly improve uterine damage in rats with experimental adenomyosis. At the same dose or half the dose of a conventional water-decoction preparation, its efficacy is superior to that of a conventional water-decoction preparation in traditional Chinese medicine. This indicates that the composition, through its innovative preparation method, has superior efficacy compared to a clinical water decoction and significant potential for use as a pharmaceutical or hospital preparation. The preparation method provided by the present invention maximizes the extraction of the composition's active ingredients, is simple to manufacture, and offers good pharmaceutical economics. DETAILED DESCRIPTION

[0030] On the one hand, the present invention provides a pharmaceutical composition for treating adenomyosis, which is composed of the following raw materials in parts by weight: 25-35 parts of Scutellaria barbata, 25-35 parts of Hedyotis diffusa, 5-15 parts of Gleditsia sinensis, 10-20 parts of Curcuma zedoaria, 5-15 parts of Eupolyphaga sinensis, 10-20 parts of Curculigo orchioides, 10-20 parts of Epimedium, and 15-25 parts of Polyporus umbellatus.

[0031] Optionally, the pharmaceutical composition is composed of the following raw materials in parts by weight: 28-32 parts of Scutellaria barbata, 28-32 parts of Hedyotis diffusa, 8-12 parts of Gleditsia sinensis, 12-18 parts of Curcuma zedoaria, 8-12 parts of Eupolyphaga sinensis, 10-20 parts of Curculigo orchioides, 12-18 parts of Epimedium, and 18-22 parts of Polyporus umbellatus.

[0032] Optionally, the pharmaceutical composition is composed of the following raw materials in parts by weight: 30 parts of Scutellaria barbata, 30 parts of Hedyotis diffusa, 10 parts of Gleditsia sinensis, 15 parts of Curcuma zedoaria, 10 parts of Eupolyphaga sinensis, 15 parts of Curculigo orchioides, 15 parts of Epimedium, and 20 parts of Polyporus umbellatus.

[0033] On the other hand, the present invention also provides a method for preparing any one of the above-mentioned pharmaceutical compositions for treating adenomyosis, comprising the following steps:

[0034] S1: Gnaphalium sylvestris and Curcuma zedoaria are crushed into coarse powder respectively to obtain Gnaphalium sylvestris powder and Curcuma zedoaria powder.

[0035] S2: placing the Gleditsia sinensis powder, Curcuma zedoaria powder, Hedyotis diffusa and Scutellaria barbata in the same extraction device, soaking them in a solid-liquid ratio of 1, and then performing reflux extraction to obtain extract 1; optionally, the solid-liquid ratio of 1 is 1g:8-12mL; the liquid used for soaking is 50% ethanol.

[0036] S3: soaking the Curculigo, Epimedium, Eupolyphaga and Polyporus according to a solid-liquid ratio of two, and then performing reflux extraction to obtain a second extract; optionally, the solid-liquid ratio of two is 1g:6-10mL; the liquid used for soaking is water.

[0037] In a specific embodiment, when soaking is performed in step S2 and step S3, the soaking time is 1-2 hours; when reflux extraction is performed, reflux extraction is performed 2-3 times, each time for 0.5-2 hours.

[0038] S4: mixing the extract 1 and the extract 2, concentrating, re-dissolving and fixing the volume to obtain the pharmaceutical composition for treating adenomyosis.

[0039] In a specific embodiment, during concentration, the solution is concentrated under reduced pressure at 60° C. until there is no alcohol taste, and then re-dissolved and fixed to volume with pure water.

[0040] In a specific embodiment, during concentration, the solution is concentrated under reduced pressure at 60° C. until there is no alcohol taste, and then re-dissolved and fixed to volume with pure water.

[0041] The Hedyotis diffusa described in the present invention is the dried whole herb of Hedyotis diffusa of the Rubiaceae family, and the plant origins of the other medicinal raw materials comply with the provisions of Part I of the 2020 edition of the "Chinese Pharmacopoeia".

[0042] It should be noted that the preparation method of the above embodiment is only a preferred preparation method of the present invention, which can extract the active ingredients of the composition, such as flavonoids, to the greatest extent, and has a simple process and good drug economy. When using the present invention, according to the components and dosage of the pharmaceutical composition, those skilled in the art can also add pharmaceutically acceptable excipients to prepare pharmaceutical preparations suitable for administration through any digestive tract route. These preparations can be prepared by combining the active ingredient with a carrier or excipient by any method known in the pharmaceutical technology, such as tablets, capsules, granules, aqueous solutions, tinctures, micropills, microspheres, dripping pills, controlled-release preparations, sustained-release preparations and other in vivo administration preparations.

[0043] Example 1 Preparation method of the pharmaceutical composition of the present invention

[0044] A pharmaceutical composition for treating adenomyosis is prepared by the following steps: respectively grinding Gleditsiae Spinae and Curcuma Rhizoma into coarse powder; weighing 30g of Scutellaria barbata, 30g of Hedyotis diffusa, 10g of Gleditsiae Spinae powder, and 15g of Curcuma Rhizoma powder, soaking them in 900mL of 50% ethanol for 1 hour, refluxing for 1 hour, filtering out the extract, repeating the extraction twice with the medicinal residue, combining the extracts, and concentrating under reduced pressure at 60°C until the alcohol taste is gone, thereby obtaining a first extract; weighing 10g of Eupolyphaga sinensis, 15g of Curculigo orchioides, 15g of Epimedium, and 20g of Polyporus umbellatus, soaking them in 600mL of water for 1 hour, refluxing for 1 hour, filtering out the extract, repeating the extraction twice with the medicinal residue, and combining the extracts to obtain a second extract. The first and second extracts are combined and concentrated under reduced pressure at 60°C to 145mL, thereby obtaining the sample of Example 1.

[0045] Comparative Example 1 Preparation of Traditional Chinese Medicine Clinical Routine Water Decoction Preparation

[0046] The Chinese medicine decoction with the same formula as Example 1 is prepared by the same method as the current clinical decoction method, and is prepared by the following steps: taking 30g of Scutellaria barbata, 30g of Hedyotis diffusa, 10g of Spina Gleditsiae, 15g of Curcuma zedoaria, 10g of Eupolyphaga sinensis, 15g of Curculigo orchioides, 15g of Epimedium, and 20g of Polyporus, totaling 145g; putting each medicinal material into a decoction pot, adding 1500mL of water and soaking for 1 hour, decocting and extracting for 1 hour, filtering the extract, repeating the extraction twice with the residue, combining the extracts, and concentrating under reduced pressure at 60°C to 145mL to obtain a sample of Comparative Example 1.

[0047] The beneficial effects of the present invention are demonstrated by the following pharmacodynamic tests.

[0048] Experimental Example 1 Effect of the Composition on Experimental Adenomyosis Rats

[0049] 1. Test sample

[0050] Example 1, dark brown liquid, 1g of extract is equivalent to 2.82g of original medicinal material.

[0051] Comparative Example 1: Dark brown liquid, 1 ml of extract is equivalent to 2.71 g of original medicinal material.

[0052] Positive control: Dienogest tablets, Bayer Weimar GmbH and Co.KG, Import Drug Registration Number: H20180090, Batch Number: WEV4VS. Indication: Treatment of endometriosis; Strength: 2 mg; Dosage: Oral, one tablet daily.

[0053] 2. Animals and Grouping

[0054] Fifty-four ICR mice (30 females and 24 males, 8 weeks old) were provided by Beijing Weitonglihua Laboratory Animal Technology Co., Ltd., production license number: SCXK(Beijing)2021-0006. They were housed in the SPF barrier system of the Experimental Animal Center of Sichuan Academy of Traditional Chinese Medicine, under conditions that met the requirements of the barrier system. Laboratory Animal Use License Number: SYXK(Sichuan)2018-100.

[0055] Seven experimental groups were set up, namely blank control group, model control group, positive control group (dienogest 0.411 mg / kg), high-dose group of Example 1 (29.8 g crude drug / kg), medium-dose group of Example 1 (14.9 g crude drug / kg), low-dose group of Example 1 (7.4 g crude drug / kg), comparative example 1 group (29.8 g crude drug / kg)

[0056] 3. Test methods

[0057] ICR mice were raised in separate cages by sex and fed a normal diet. They were quarantined and observed for 3 days. On the 6th day, the animals that failed the quarantine were removed and the mice were separated again, with 2 females and 1 male per cage. After about 3 weeks of birth, the male suckling mice were removed. Except for some female suckling mice reserved as normal controls, the rest were drip-fed with 10 u The model was established by administering 1 mg / mouse tamoxifen emulsion (0.225 mg / ml) once daily for 4 consecutive days. The mice were breastfed for the first 21 days, separated from their mothers after 21 days, and fed normally for 3 months. The animals were then randomly divided into 7 groups (10-12 mice per group) according to body weight.

[0058] The drugs were administered continuously for 90 days. The body weight was measured 24 hours after the last administration, blood was collected from the orbits, and the animals were killed by cervical dislocation. The serum was separated for the detection of biochemical indicators. The uterus was obtained after dissection, weighed, and fixed in 4% neutral formaldehyde for histopathological observation.

[0059] 4. Test results

[0060] 4.1 Pathological examination

[0061] The criteria for judging uterine tissue damage are shown in Table 1.

[0062] Table 1 Uterine tissue damage score

[0063] Grading Pathological characteristics 0 Normal uterus 1 Endometrial stroma invades the muscular lining 2 Endometrial stroma and glands invade the muscular lining 3 Endometrial tissue invades the junction of the inner and outer myometrium 4 Endometrial glandular cystic hyperplasia with subserosal nodules

[0064] The test results are shown in Table 2.

[0065] Table 2 Statistical results of uterine tissue damage scores (M(Q1, Q3))

[0066]

[0067]

[0068] Note: Compared with the model control group, *P<0.05; compared with the comparative example 1 group, △P<0.05

[0069] The above results show that the uterine injury score of the animals in the model control group was significantly increased, and the difference was statistically significant (P<0.05), indicating that the model was successfully replicated; compared with the model control group, the scores of the large and medium dose groups of Example 1 were reduced, and the difference was statistically significant (P<0.05); compared with the comparative example 1 group, the scores of the large and medium dose groups of Example 1 were reduced, and the difference was statistically significant (P<0.05). Comparative example 1 did not show efficacy, which may be due to the species difference between animals and humans, resulting in a low dosage.

[0070] The above results suggest that Example 1 can significantly improve the uterine pathological manifestations of experimental adenomyosis, and its effect is better than that of Comparative Example 1 of conventional water decoction in traditional Chinese medicine clinical practice at the same dose and 1 / 2 dose, indicating that Example 1 greatly improves the efficacy through the innovation of the preparation method.

Claims

1. A method for preparing a pharmaceutical composition for treating adenomyosis, characterized in that: It includes the following steps: a. Weigh the API in the appropriate weight ratio: 25-35 parts of Scutellaria barbata, 25-35 parts of Hedyotis diffusa, 5-15 parts of Spina Gleditsiae, 10-20 parts of Curcuma zedoaria, 5-15 parts of Eupolyphaga sinensis, 10-20 parts of Curculigo orchioides, 10-20 parts of Epimedium, and 15-25 parts of Polyporus; b. Extraction of Gleditsia sinensis, Curcuma zedoaria, Hedyotis diffusa and Scutellaria barbata with organic solvents; c. Curculigo, Epimedium, Eupolyphaga sinensis, and Polyporus umbellatus are extracted with water; d. Combine the extracts from step b and step c, concentrate them, add pharmaceutically acceptable excipients or auxiliary ingredients, and prepare a commonly used pharmaceutical preparation.

2. The method for preparing the pharmaceutical composition for treating adenomyosis according to claim 1, wherein: The weight ratio of the raw material medicine described in step a is: 28-32 parts of Scutellaria barbata, 28-32 parts of Hedyotis diffusa, 8-12 parts of Spina Gleditsiae, 12-18 parts of Curcuma zedoaria, 8-12 parts of Eupolyphaga sinensis, 10-20 parts of Curculigo orchioides, 12-18 parts of Epimedium, and 18-22 parts of Polyporus.

3. The method for preparing the pharmaceutical composition for treating adenomyosis according to claim 2, wherein: The weight ratio of the raw material medicine described in step a is: 30 parts of Scutellaria barbata, 30 parts of Herba Hedyotis diffusae, 10 parts of Spina Gleditsiae, 15 parts of Rhizoma Curcumae, 10 parts of Eupolyphaga Seu Steleophora, 15 parts of Curculigo orchioides, 15 parts of Herba Epimedii, 20 parts of Polyporus.

4. The method for preparing the pharmaceutical composition for treating adenomyosis according to any one of claims 1 to 3, wherein: The organic solvent in step b is ethanol, methanol, acetone, petroleum ether, benzene, chloroform, ether, ethyl acetate, or dichloroethane; preferably, the organic solvent is ethanol; The extraction method described in step b is first leaching, then reflux extraction and ultrasonic extraction.

5. The method for preparing the pharmaceutical composition for treating adenomyosis according to claim 4, characterized in that: The ethanol is 40-90% ethanol, preferably, the ethanol is 50% ethanol; the material-liquid ratio of the medicinal material to the ethanol is 1g:8-12mL.

6. The method for preparing the pharmaceutical composition for treating adenomyosis according to any one of claims 1 to 3, characterized in that: The extraction method in step c is first leaching, then reflux extraction or ultrasonic extraction; The material-liquid ratio of medicinal materials to water is: 1g:6-10mL.

7. The method for preparing the pharmaceutical composition for treating adenomyosis according to any one of claims 4 to 6, characterized in that: The soaking time is 1-2 hours; The reflux extraction is performed 2-3 times, each time for 0.5-2 hours.

8. The method for preparing the pharmaceutical composition for treating adenomyosis according to claim 1, wherein: The concentration method in step d is reduced pressure concentration, and the concentration temperature is 60°C.

9. The method for preparing the pharmaceutical composition for treating adenomyosis according to claim 1, wherein: The preparations are tablets, capsules, granules, aqueous solutions, tinctures, micropills, microspheres, dripping pills, controlled-release preparations, and sustained-release preparations.

Images