Traditional Chinese medicine composition for treating benign prostatic hyperplasia, medicinal preparation and preparation method thereof

Abstract

The invention provides a traditional Chinese medicine composition for treating benign prostatic hyperplasia, a medicinal preparation and a preparation method thereof, and belongs to the technical field of medicines. The traditional Chinese medicine composition comprises the following components: seaweed, kelp, cuttlebone, yam rhizome, salvia miltiorrhiza, radix scrophulariae, dandelion, semen cuscutae and semen plantaginis. The traditional Chinese medicine composition and the medicinal preparation provided by the invention have the effects of removing stasis, removing stasis, promoting diuresis and removing turbidity, can be used for treating chronic prostatitis damp-heat stasis syndrome and prostatic hyperplasia with nodule formation, and are obvious in damp-heat stasis resistance.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a traditional Chinese medicine composition, pharmaceutical preparation and preparation method for treating benign prostatic hyperplasia. Background Technology

[0002] Benign prostatic hyperplasia (BPH), also known as enlarged prostate, is a disease caused by hormonal imbalances and other factors. It results in the gradual proliferation of connective tissue and smooth muscle tissue in the middle or concave lobes of the posterior urethra, forming multiple globular nodules and causing a series of functional disorders in the urethra, bladder, and kidneys. The symptoms of BPH mainly manifest in two ways: one is bladder irritation symptoms, primarily manifested as urinary frequency, urgency, nocturia, and urge incontinence; the other is obstructive symptoms caused by the enlarged prostate obstructing the urinary tract.

[0003] Western medicine treatment for benign prostatic hyperplasia (BPH) often involves the use of hormones or anti-hormonal drugs, alpha-adrenergic receptor blockers, 5α-reductase inhibitors, and cholesterol inhibitors. Its disadvantages include significant side effects, a high risk of dependence, and long-term use can cause considerable harm to the cardiovascular and immune systems. Furthermore, because Western medicine does not fully understand the root causes and pathogenesis of BPH, its treatments primarily focus on improving and alleviating symptoms rather than curing the condition.

[0004] In Traditional Chinese Medicine (TCM), benign prostatic hyperplasia (BPH) is classified as damp-heat stagnation syndrome. TCM treatments for BPH vary. For example, Qianliekangshu Capsules contain ingredients such as Smilax glabra, Polygonum cuspidatum, Carapax Trionycis, Curcuma zedoaria, Epimedium, Astragalus membranaceus, and Lycium barbarum. Its effects focus on detoxification, blood circulation, and tonifying the kidneys, particularly addressing kidney deficiency and damp-heat stagnation. It is suitable for chronic prostatitis with kidney deficiency and damp-heat syndrome (frequent urination, urgency, lower back and knee weakness, and testicular pain). This medicine combines tonification and elimination: Carapax Trionycis and Curcuma zedoaria resolve stagnation and dissipate nodules, while Astragalus membranaceus and Lycium barbarum tonify the kidneys and replenish qi. It is suitable for chronic inflammation with a long course. Qingzhuo Qudu Pills contain ingredients such as Phellodendron chinense, Polygonum aviculare, Plantago asiatica, and Poria cocos. Its effects focus on clearing heat and dampness, resolving stagnation, and promoting urination. This medicine primarily targets damp-heat in the lower burner; it is suitable for symptoms of damp-heat accumulation in the lower burner, such as prostatitis or urinary tract infections (frequent and urgent urination, urethral burning, scrotal dampness); this medicine specifically targets damp-heat: its ingredients mostly contain diuretics and urinary tract clearing agents, providing significant short-term improvement in urination symptoms. Bi Xie Fen Qing Yin (from *Medical Insights*) contains Bi Xie, Plantago seed, Poria, Atractylodes macrocephala, and Acorus tatarinowii; its effects focus on eliminating dampness and turbidity; it is suitable for symptoms of damp-heat accumulation in the lower burner, such as urinary tract infections (turbid urine, frequent and painful urination); this medicine is ineffective against "tangible nodules" formed by glandular hyperplasia and lacks kidney-tonifying and foundation-strengthening effects.

[0005] Since the causes and symptoms of benign prostatic hyperplasia (BPH) vary, and different Chinese herbal medicines have different focuses in their efficacy, more Chinese herbal medicines for treating BPH still need to be developed in order to better address different types of the disease. Summary of the Invention

[0006] To address the shortcomings of existing technologies, the present invention aims to provide a traditional Chinese medicine composition, pharmaceutical preparation, and preparation method for treating benign prostatic hyperplasia (BPH). This traditional Chinese medicine composition and pharmaceutical preparation possess the effects of dispersing nodules and removing blood stasis, as well as promoting diuresis and eliminating turbidity. It can be used to treat chronic prostatitis with damp-heat stagnation syndrome, and BPH accompanied by nodule formation and significant damp-heat stagnation.

[0007] To achieve this objective, the present invention adopts the following technical solution:

[0008] In a first aspect, the present invention provides a traditional Chinese medicine composition for treating benign prostatic hyperplasia, the traditional Chinese medicine composition comprising the following components:

[0009] Seaweed, kelp, cuttlebone, Dioscorea hypoglauca, Salvia miltiorrhiza, Scrophularia ningpoensis, dandelion, Cuscuta chinensis, and Plantago asiatica.

[0010] In some preferred embodiments of the present invention, the traditional Chinese medicine composition comprises the following components in parts by weight:

[0011] Seaweed 12-18 parts, kelp 12-18 parts, cuttlebone 18-22 parts, Dioscorea hypoglauca 28-32 parts, Salvia miltiorrhiza 18-22 parts, Scrophularia ningpoensis 28-32 parts, dandelion 22-26 parts, Cuscuta chinensis 18-22 parts, and Plantago asiatica 12-18 parts.

[0012] The weight percentage of seaweed can be any value within the range of 12-18 parts, such as 12 parts, 12.2 parts, 12.5 parts, 12.8 parts, 13 parts, 13.2 parts, 13.5 parts, 13.8 parts, 14 parts, 14.2 parts, 14.5 parts, 14.8 parts, 15 parts, 15.2 parts, 15.5 parts, 15.8 parts, 16 parts, 16.2 parts, 16.5 parts, 16.8 parts, 17 parts, 17.2 parts, 17.5 parts, 17.8 parts, or 18 parts, etc.

[0013] The weight of kelp can be any value within the range of 12-18 parts, such as 12 parts, 12.2 parts, 12.5 parts, 12.8 parts, 13 parts, 13.2 parts, 13.5 parts, 13.8 parts, 14 parts, 14.2 parts, 14.5 parts, 14.8 parts, 15 parts, 15.2 parts, 15.5 parts, 15.8 parts, 16 parts, 16.2 parts, 16.5 parts, 16.8 parts, 17 parts, 17.2 parts, 17.5 parts, 17.8 parts, or 18 parts, etc.

[0014] The weight of cuttlebone can be any value within the range of 18-22 parts, such as 18 parts, 18.2 parts, 18.5 parts, 18.8 parts, 19 parts, 19.2 parts, 19.5 parts, 19.8 parts, 20 parts, 20.2 parts, 20.5 parts, 20.8 parts, 21 parts, 21.2 parts, 21.5 parts, 21.8 parts, or 22 parts, etc.

[0015] The weight of Dioscorea hypoglauca can be any value within the range of 28-32 parts, such as 28 parts, 28.2 parts, 28.5 parts, 28.8 parts, 29 parts, 29.2 parts, 29.5 parts, 29.8 parts, 30 parts, 30.2 parts, 30.5 parts, 30.8 parts, 31 parts, 31.2 parts, 31.5 parts, 31.8 parts, or 32 parts, etc.

[0016] The weight of Danshen can be any value within the range of 18-22 parts, such as 18 parts, 18.2 parts, 18.5 parts, 18.8 parts, 19 parts, 19.2 parts, 19.5 parts, 19.8 parts, 20 parts, 20.2 parts, 20.5 parts, 20.8 parts, 21 parts, 21.2 parts, 21.5 parts, 21.8 parts, or 22 parts, etc.

[0017] The weight of Scrophularia can be any value within the range of 28-32 parts, such as 28 parts, 28.2 parts, 28.5 parts, 28.8 parts, 29 parts, 29.2 parts, 29.5 parts, 29.8 parts, 30 parts, 30.2 parts, 30.5 parts, 30.8 parts, 31 parts, 31.2 parts, 31.5 parts, 31.8 parts, or 32 parts, etc.

[0018] The weight of dandelion can be any value within the range of 22-26 parts, such as 22 parts, 22.2 parts, 22.5 parts, 22.8 parts, 23 parts, 23.2 parts, 23.5 parts, 23.8 parts, 24 parts, 24.2 parts, 24.5 parts, 24.8 parts, 25 parts, 25.2 parts, 25.5 parts, 25.8 parts, or 26 parts, etc.

[0019] The weight of dodder seeds can be any value within the range of 18-22 parts, such as 18 parts, 18.2 parts, 18.5 parts, 18.8 parts, 19 parts, 19.2 parts, 19.5 parts, 19.8 parts, 20 parts, 20.2 parts, 20.5 parts, 20.8 parts, 21 parts, 21.2 parts, 21.5 parts, 21.8 parts, or 22 parts, etc.

[0020] The weight percentage of plantain seeds can be any value within the range of 12-18 parts, such as 12 parts, 12.2 parts, 12.5 parts, 12.8 parts, 13 parts, 13.2 parts, 13.5 parts, 13.8 parts, 14 parts, 14.2 parts, 14.5 parts, 14.8 parts, 15 parts, 15.2 parts, 15.5 parts, 15.8 parts, 16 parts, 16.2 parts, 16.5 parts, 16.8 parts, 17 parts, 17.2 parts, 17.5 parts, 17.8 parts, or 18 parts, etc.

[0021] In some preferred embodiments of the present invention, the traditional Chinese medicine composition comprises the following components in parts by weight:

[0022] Seaweed 15 parts, kelp 15 parts, cuttlebone 20 parts, Dioscorea hypoglauca 30 parts, Salvia miltiorrhiza 20 parts, Scrophularia ningpoensis 30 parts, dandelion 24 parts, Cuscuta chinensis 20 parts, and Plantago asiatica 15 parts.

[0023] The efficacy of each component in the traditional Chinese medicine composition provided by this invention is as follows:

[0024] Seaweed: It can resolve phlegm, soften hard masses, promote diuresis, and dissipate nodules;

[0025] Kelp: softens and disperses nodules, promotes diuresis and reduces swelling;

[0026] Cuttlebone: Astringent, hemostatic, antacid, and analgesic;

[0027] Dioscorea hypoglauca: promotes diuresis, eliminates turbidity, relieves urinary tract infections and pain;

[0028] Danshen: Promotes blood circulation, removes blood stasis, regulates menstruation, and relieves pain;

[0029] Scrophularia: Clears heat and cools blood, nourishes yin and detoxifies;

[0030] Dandelion: Clears heat and detoxifies, reduces swelling and dissipates nodules;

[0031] Cuscuta: Tonifies the kidneys and strengthens essence, benefits the liver and improves eyesight;

[0032] Plantain seed: Clears heat and promotes diuresis, eliminates dampness and relieves strangury.

[0033] The compatibility mechanism of each component is as follows:

[0034] The principal herbs are: Dioscorea hypoglauca, Sargassum, and Laminaria japonica. Dioscorea hypoglauca has the effects of promoting diuresis and removing turbidity, and separating clear from turbid fluids, specifically targeting the main symptoms of damp-heat accumulation in prostatitis (frequent urination, urgency, and urethral burning). Sargassum and Laminaria japonica have the effects of softening and dispersing nodules, and resolving phlegm and masses, targeting the pathological basis of prostatic hyperplasia, fibrosis, or nodule formation. The three herbs work synergistically to both clear damp-heat to treat the symptoms and resolve phlegm to improve local obstruction, embodying the principle of "treating both the root cause and the symptoms."

[0035] Assistant herbs: Plantain seed, dandelion, and salvia miltiorrhiza. Plantain seed promotes urination and relieves strangury, assists Dioscorea hypoglauca in expelling damp-heat, and guides the medicine downward to the lower abdomen. Dandelion clears heat and detoxifies, inhibits bacterial infection, and reduces redness, swelling, heat, and pain. Salvia miltiorrhiza promotes blood circulation and removes blood stasis, improves prostate microcirculation, and relieves congestion, pain, and tissue fibrosis. These three herbs assist the principal herbs from the perspectives of promoting diuresis, clearing heat, and promoting blood circulation, respectively, to enhance the overall therapeutic effect.

[0036] Adjuvant herbs: Scrophularia, Cuscuta, and Cuscuta chinensis. Scrophularia nourishes yin and cools the blood, preventing prolonged damp-heat from damaging yin, and also clears deficiency fire. Cuscuta chinensis tonifies the kidneys and strengthens essence, supporting vital energy, and is effective for prostatitis accompanied by sexual dysfunction or kidney deficiency and lower back pain. Cuscuta chinensis astringes and consolidates, relieving symptoms of frequent and urgent urination, and repairing the urethral mucosa. Scrophularia counteracts the cold and dryness of heat-clearing and dampness-draining herbs, Cuscuta chinensis tonifies deficiency and strengthens the body's foundation, and Cuscuta chinensis harmonizes opening and closing, embodying a "combination of attack and tonification" approach.

[0037] Guided drug: Plantago seed also serves as a guide drug, promoting urination and relieving strangury while guiding other drugs directly to the affected area in the lower abdomen (prostate and urinary system), ensuring that the medicinal power is concentrated on the target point and enhancing local efficacy.

[0038] The traditional Chinese medicine composition provided by this invention is based on the core principles of "softening and dispersing nodules, promoting diuresis and relieving strangury". Targeting the pathogenesis of prostatitis characterized by "damp-heat stagnation, phlegm accumulation and kidney deficiency", it combines Dioscorea hypoglauca, Sargassum, and Laminaria japonica to promote diuresis, resolve phlegm, and dissipate nodules to improve glandular edema and fibrosis. It is supplemented with Salvia miltiorrhiza to invigorate blood and remove blood stasis, and Taraxacum mongolicum to clear heat and detoxify, directly relieving inflammation and pain. It is further supplemented with Scrophularia ningpoensis to nourish yin and cool blood, and Cuscuta chinensis to tonify the kidney and strengthen the foundation, harmonizing the cold and dryness of the purgative drugs, and addressing the root cause of yin deficiency due to prolonged illness. Plantago asiatica guides the medicine downward, and Cuscuta chinensis astringes and consolidates, treating both the symptoms and the root cause.

[0039] This traditional Chinese medicine composition works synergistically to dispel stagnation and remove blood stasis, as well as to eliminate dampness and turbidity (primarily by softening and dispersing stagnation and clearing damp heat, while also promoting blood circulation, removing blood stasis, and tonifying the kidneys). It can be used to treat chronic prostatitis with damp-heat stagnation syndrome (common symptoms include difficulty urinating, frequent urination, and urgency), and benign prostatic hyperplasia with nodule formation and significant damp-heat stagnation (such as painful urination, difficulty urinating, perineal distending pain, yellow and greasy tongue coating, and wiry and hesitant pulse).

[0040] In a second aspect, the present invention provides a pharmaceutical preparation for treating benign prostatic hyperplasia, the pharmaceutical preparation comprising the effective components of the traditional Chinese medicine composition as described in the first aspect.

[0041] In some embodiments of the present invention, the pharmaceutical preparation further includes excipients.

[0042] In some embodiments of the present invention, the dosage form of the pharmaceutical preparation is granules, tablets, capsules, or decoction.

[0043] Thirdly, the present invention provides a method for preparing a pharmaceutical preparation as described in the second aspect, the method comprising the following steps:

[0044] (1) The traditional Chinese medicine composition described in the first aspect is subjected to water extraction to obtain an aqueous extract;

[0045] (2) The aqueous extract is prepared into the dosage form of the drug preparation.

[0046] In some embodiments of the present invention, the water extraction step includes:

[0047] The herbal composition is decocted with water at least twice (e.g., twice, three times, four times, or five times), each time for 1-4 hours (e.g., 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 3.5 hours, or 4 hours). After each decoction, the mixture is filtered, and the filtrates are combined to obtain the aqueous extract.

[0048] In some embodiments of the present invention, in the water extraction step, the total mass of water is 18-35 times the mass of the traditional Chinese medicine composition; for example, it can be 18 times, 19 times, 20 times, 22 times, 23 times, 25 times, 26 times, 28 times, 30 times, 32 times, 33 times or 35 times, etc.

[0049] In some embodiments of the present invention, the water extraction step involves two decoctions. For the first decoction, the mass of water is 10-20 times the mass of the herbal composition (e.g., 10, 12, 13, 15, 16, 18, or 20 times, etc.), and the decoction time is 2-3 hours (e.g., 2 hours, 2.2 hours, 2.3 hours, 2.5 hours, 2.6 hours, 2.8 hours, or 3 hours, etc.). For the second decoction, the mass of water is 8-15 times the mass of the herbal composition (e.g., 8, 9, 10, 11, 12, 13, 14, or 15 times, etc.), and the decoction time is 1-2 hours (e.g., 1 hour, 1.2 hours, 1.3 hours, 1.5 hours, 1.6 hours, 1.8 hours, or 2 hours, etc.).

[0050] Compared with the prior art, the present invention has the following beneficial effects:

[0051] This invention utilizes a scientific combination of seaweed, kelp, cuttlebone, dioscorea hypoglauca, salvia miltiorrhiza, scrophularia ningpoensis, dandelion, cuscuta chinensis, and plantain seed to obtain a traditional Chinese medicine composition and preparation with the effects of dispersing nodules and removing blood stasis, and promoting diuresis and eliminating turbidity (mainly softening and dispersing nodules and clearing damp heat, while also promoting blood circulation and removing blood stasis and tonifying the kidney and strengthening the foundation). It can be used to treat chronic prostatitis with damp-heat stagnation syndrome (common symptoms include difficulty urinating, frequent urination, and urgency), benign prostatic hyperplasia with nodule formation and significant damp-heat stagnation (such as painful urination, difficulty urinating, perineal distending pain, yellow and greasy tongue coating, and wiry and hesitant pulse). Attached Figure Description

[0052] Figure 1 A bar chart showing the prostate coefficients of rats in each group;

[0053] Figure 2A Bar chart showing PCAP content in prostate tissue of rats in each group;

[0054] Figure 2B Bar chart showing serum E2 levels in rats of each group;

[0055] Figure 2C Bar chart showing serum DHT levels in rats of each group;

[0056] Figure 2D A bar chart showing the serum T content of rats in each group;

[0057] Figure 3 HE staining images of prostate tissue from rats in each group. Detailed Implementation

[0058] The technical solution of the present invention will be further described below with reference to the accompanying drawings and specific embodiments. Those skilled in the art should understand that the specific embodiments described are merely illustrative of the present invention and should not be construed as limiting the invention.

[0059] Example 1

[0060] This embodiment provides a traditional Chinese medicine composition for treating benign prostatic hyperplasia, comprising the following components in parts by weight:

[0061] Seaweed 12 parts, kelp 18 parts, cuttlebone 18 parts, Dioscorea hypoglauca 32 parts, Salvia miltiorrhiza 18 parts, Scrophularia ningpoensis 32 parts, dandelion 22 parts, Cuscuta chinensis 22 parts, and Plantago asiatica 12 parts.

[0062] This embodiment also provides a decoction for treating benign prostatic hyperplasia, the preparation method of which is as follows:

[0063] Weigh the medicinal materials according to the proportions of the Chinese herbal composition, add water and decoct twice. For the first decoction, add 20 times the amount of water as the medicinal materials and decoct for 3 hours. For the second decoction, add 15 times the amount of water as the medicinal materials and decoct for 2 hours. After each decoction, filter, combine the filtrates, and concentrate under reduced pressure to obtain a decoction for treating benign prostatic hyperplasia.

[0064] Example 2

[0065] This embodiment provides a traditional Chinese medicine composition for treating benign prostatic hyperplasia, comprising the following components in parts by weight:

[0066] Seaweed 18 parts, kelp 12 parts, cuttlebone 22 parts, Dioscorea hypoglauca 28 parts, Salvia miltiorrhiza 22 parts, Scrophularia ningpoensis 28 parts, dandelion 26 parts, Cuscuta chinensis 18 parts, and Plantago asiatica 18 parts.

[0067] This embodiment also provides a decoction for treating benign prostatic hyperplasia, the preparation method of which is as follows:

[0068] Weigh the medicinal materials according to the proportions of the Chinese herbal composition, add water and decoct twice. For the first decoction, add 10 times the amount of water as the medicinal materials and decoct for 2 hours. For the second decoction, add 8 times the amount of water as the medicinal materials and decoct for 1 hour. After each decoction, filter, combine the filtrates, and concentrate under reduced pressure to obtain a decoction for treating benign prostatic hyperplasia.

[0069] Example 3

[0070] This embodiment provides a traditional Chinese medicine composition for treating benign prostatic hyperplasia, comprising the following components in parts by weight:

[0071] Seaweed 15 parts, kelp 15 parts, cuttlebone 20 parts, Dioscorea hypoglauca 30 parts, Salvia miltiorrhiza 20 parts, Scrophularia ningpoensis 30 parts, dandelion 24 parts, Cuscuta chinensis 20 parts, and Plantago asiatica 15 parts.

[0072] This embodiment also provides a decoction for treating benign prostatic hyperplasia, and its preparation method is as follows:

[0073] Weigh the medicinal materials according to the ratio of the traditional Chinese medicine composition, decoct them with water twice. For the first decoction, add 15 times the mass of the medicinal materials in water and decoct for 2.5 hours; for the second decoction, add 10 times the mass of the medicinal materials in water and decoct for 1.5 hours; filter after each decoction, combine the filtrates, and concentrate them under reduced pressure to 2.46 g of the traditional Chinese medicine composition per mL to obtain the decoction for treating benign prostatic hyperplasia, and store it at 4°C for later use.

[0074] Therapeutic efficacy test

[0075] Animal experiments were conducted using the decoction for treating benign prostatic hyperplasia prepared in Example 3 to confirm its therapeutic effect on the rat model of benign prostatic hyperplasia.

[0076] 1. Test materials

[0077] 1.1. Test animals

[0078] SPF-grade male SD rats, weighing 240 g - 250 g, were purchased from Spf (Suzhou) Biotechnology Co., Ltd., with the production license number SCXK (Su) 2022 - 0006 and the certificate number A202410080022. Feeding environmental conditions: The feeding environmental conditions are standard, referring to the national standard GB14925 - 2010 of the People's Republic of China; the temperature is 20 - 26°C (daily temperature difference ≤ 4°C); the relative humidity is 40 - 70%; the pressure in the feeding room ≥ 10 Pa; 12 / 12 hours of day and night light and dark alternation.

[0079] 1.2. Test reagents

[0080] The decoction for treating benign prostatic hyperplasia prepared in Example 3 was diluted with sterile injection water to 4.92 g / 8 mL, 9.85 g / 8 mL, and 19.69 g / 8 mL respectively. This concentration refers to the mass of the traditional Chinese medicine composition corresponding to the decoction contained in each 8 mL of the solution.

[0081] Wenglitong capsules, with a specification of 0.4 g per capsule, batch number 240602, stored in a sealed manner, produced by Jilin Huawei Pharmaceutical Co., Ltd.; the product approval number is国药准字 Z20043767. Preparation method: Dilute the content of Wenglitong capsules with a 0.5% sodium carboxymethylcellulose solution to 0.13 g / 8 mL.

[0082] Testosterone propionate injection, with a specification of 1 mL: 25 mg per vial, batch number of 80131241, stored in a sealed manner, produced by Sichuan Jingke Biochemical Products Co., Ltd.; the product approval number is兽药字 220411054.

[0083] Sodium carboxymethyl cellulose, 500g / bottle, batch number 20210402, store in a tightly closed container, manufactured by Sinopharm Chemical Reagent Co., Ltd.; product number 30036328. Preparation method: Dilute sodium carboxymethyl cellulose to 0.5wt% with sterile water for injection.

[0084] Sterile water for injection, 500 mL / bottle, batch number 240621, to be stored in a sealed container, manufactured by Shaanxi Sheng'ao Animal Pharmaceutical Co., Ltd.; product approval number is Veterinary Drug No. 270071791.

[0085] This is a general-purpose tissue fixative, available in 500mL vials, batch number GP24103081634. Store at room temperature. The manufacturer is Wuhan Saiweier Biotechnology Co., Ltd.

[0086] 1.3. Instruments

[0087] The pipette is manufactured by Eppendorf and has a model number of 200μL.

[0088] The electronic balance is manufactured by Shanghai Yaoxin Electronic Technology Co., Ltd., and its model number is LQ-C12001.

[0089] High-speed low-temperature tissue homogenizer, manufactured by Wuhan Saiweier Biotechnology Co., Ltd., model KZ-1111-F;

[0090] The benchtop high-speed refrigerated microcentrifuge is manufactured by Thermo and its model number is Sorvall Legand Micro 17R.

[0091] Mini mixing centrifuge, manufacturer: LABGIC, model: L-CM-MINI;

[0092] The microplate reader is manufactured by Thermo and is model VARIOSKAN LUX.

[0093] The dehydrator is manufactured by DIAPATH and its model is Donatello.

[0094] The embedding machine is manufactured by Wuhan Junjie Electronics Co., Ltd., and its model number is JB-P5.

[0095] The freezing station is manufactured by Wuhan Junjie Electronics Co., Ltd., and its model number is JB-L5.

[0096] The pathology slider is manufactured by Leica Instruments Shanghai Co., Ltd., and its model number is RM2016.

[0097] The sheet spreader is manufactured by Zhejiang Jinhua Kedi Instrument Equipment Co., Ltd., and its model is KD-P.

[0098] The oven is manufactured by Tianjin Laiborui Instrument Equipment Co., Ltd., and its model is GFL-230.

[0099] An upright optical microscope, manufactured by Nikon Japan, model Nikon Eclipse E100;

[0100] The imaging system is manufactured by Nikon of Japan, and its model is NIKON DS-U3.

[0101] An upright white light photographic microscope, manufactured by Nikon Japan, model Eclipse Ci-L.

[0102] 2. Test Methods

[0103] 2.1. Modeling, grouping, and drug administration

[0104] After being fed a normal diet for one week, the rats were randomly divided into 6 groups: blank group, model group, low-dose prescription group, medium-dose prescription group, high-dose prescription group, and Wenglitong capsule group, with 10 rats in each group.

[0105] After weighing, rats were anesthetized by intraperitoneal injection of 10% chloral hydrate (0.3 mL / 100 g). Routine disinfection and antibiotic injections were administered to prevent infection. In all groups except the control group, bilateral testes were removed from male rats under aseptic conditions, the stumps were ligated, and the skin was sutured for subsequent experiments. In the control group, the scrotum was incised and then sutured, without removal of the testes, thus undergoing a sham surgery. A small amount of povidone-iodine was applied to the wounds for the first three days post-surgery. After one week of natural recovery, all rats except the control group were subcutaneously injected with testosterone propionate (5 mg / kg) on ​​the eighth day post-surgery to induce a rat model of benign prostatic hyperplasia (BPH).

[0106] After successful modeling, each group of rats was administered the drug twice daily by gavage for 30 consecutive days. The types and dosages of drugs administered to each group of rats are as follows:

[0107] Blank group and model group: physiological saline, administration volume 800μL / 100g;

[0108] Low-dose prescription group: a diluted solution of the decoction prepared in Example 3 with a concentration of 4.92 g / 8 mL, administered at a dose of 4.92 g / kg, with an administration volume of 800 μL / 100 g;

[0109] The dosage group in the prescription was a diluted solution of the decoction prepared in Example 3 with a concentration of 9.85 g / 8 mL, with a dosage of 9.85 g / kg and a dosage volume of 800 μL / 100 g.

[0110] High-dose prescription group: a diluted solution of the decoction prepared in Example 3 with a concentration of 19.69 g / 8 mL, administered at a dose of 19.69 g / kg and a volume of 800 μL / 100 g;

[0111] Wenglitong capsule group: dilution of Wenglitong capsules with a concentration of 0.13g / 8mL, dosage of 0.13g / kg, and administration volume of 800μL / 100g.

[0112] 2.2. Detection Indicators and Methods

[0113] 2.2.1. Prostate coefficient

[0114] The rats were weighed, and after the rats were sacrificed, the prostate was weighed. The prostate coefficient was calculated using the following formula: Prostate coefficient = Prostate weight / Body weight × 100%.

[0115] 2.2.2. Serum T, DHT, and E2 levels

[0116] The detection was performed using an ELISA kit, and the steps are as follows:

[0117] (1) Sample collection: Use test tubes free of pyrogens and endotoxins, and avoid any cell stimulation during the operation. After the experiment, take rat blood and centrifuge at 3000 rpm for 10 minutes to quickly and carefully separate serum and red blood cells.

[0118] (2) Remove the required strips from the aluminum foil bag after equilibration at room temperature for 20 minutes, and seal the remaining strips in a self-sealing bag and return them to 4℃.

[0119] (3) Set up standard wells and sample wells, and add 50 μL of standard of different concentrations to each standard well.

[0120] (4) Add 10 μL of the sample to be tested to the sample well first, and then add 40 μL of sample diluent; do not add to the blank well.

[0121] (5) Except for the blank wells, add 100 μL of horseradish peroxidase (HRP) labeled detection antibody to each of the standard wells and sample wells, seal the reaction wells with sealing film, and incubate at 37°C in a water bath or incubator for 60 min.

[0122] (6) Discard the liquid, pat dry on absorbent paper, fill each well with washing liquid, let stand for 1 minute, shake off the washing liquid, pat dry on absorbent paper, and repeat the washing process 5 times (or use a plate washer).

[0123] (7) Add 50 μL of substrate A and B to each well and incubate at 37°C in the dark for 15 min.

[0124] (8) Add 50 μL of stop solution to each well and measure the OD value of each well at a wavelength of 450 nm within 15 min.

[0125] (9) Plot the standard curve: plot the linear regression curve of the standard with the concentration of the standard on the x-axis and the corresponding OD value on the y-axis, and calculate the concentration value of each sample according to the curve equation.

[0126] 2.2.3. Prostatic acid phosphatase (PACP) content

[0127] Add 100 mg of prostate tissue to 1 mL of physiological saline and crush. Centrifuge at 3000 rpm for 10 minutes and collect the supernatant. Detect according to the ELISA kit instructions, see section 2.2.2.

[0128] 2.2.4. Prostate HE staining

[0129] (1) Paraffin slice preparation

[0130] 1) Sample collection: After the experiment, rats were euthanized, and prostate tissue was collected and fixed with fixative for at least 24 hours. The tissue was removed from the fixative and trimmed in a fume hood using a scalpel. The trimmed tissue and corresponding labels were then placed in a dehydration box.

[0131] 2) Dehydration and wax impregnation: The dehydration box is placed in a dehydrator for gradient alcohol dehydration. The steps are as follows: 75% alcohol for 4 hours, 85% alcohol for 2 hours, 90% alcohol for 2 hours, 95% alcohol for 1 hour, anhydrous ethanol I for 30 minutes, anhydrous ethanol II for 30 minutes, benzyl alcohol for 10 minutes, xylene I for 10 minutes, xylene II for 10 minutes, 65℃ melted paraffin I for 1 hour, 65℃ melted paraffin II for 1 hour, and 65℃ melted paraffin III for 1 hour.

[0132] 3) Embedding: The paraffin-impregnated tissue is embedded in an embedding machine. First, the molten paraffin is placed into the embedding frame. Before the paraffin solidifies, the tissue is removed from the dehydration box, placed into the embedding frame according to the embedding surface requirements, and labeled accordingly. The tissue is then cooled on a -20°C freezing stage. After the paraffin solidifies, the paraffin block is removed from the embedding frame and trimmed.

[0133] 4) Sectioning: Place the trimmed wax block on a -20℃ freezing stage to cool, then place the cooled wax block on a paraffin microtome to section to a thickness of 4μm. Float the sections on 40℃ warm water in a slide to flatten the tissue, then lift the tissue onto a glass slide and bake in a 60℃ oven. After the wax has melted in the water, remove the slide and store it at room temperature for later use.

[0134] (2) Staining

[0135] 1) Dewaxing paraffin sections to water: Place the sections in xylene I for 20 min, xylene II for 20 min, anhydrous ethanol I for 5 min, anhydrous ethanol II for 5 min, 75% ethanol for 5 min, and then wash with tap water.

[0136] 2) Hematoxylin staining: After dewaxing, the sections were stained with hematoxylin solution for 5 minutes, washed with tap water, differentiated with differentiation solution, washed with tap water, blued with blue solution, and rinsed with running water.

[0137] 3) Eosin staining: After hematoxylin staining, the sections were dehydrated in 85% and 95% alcohol for 5 minutes each, and then stained in eosin staining solution for 5 minutes.

[0138] 4) Dehydration and mounting: After eosin staining, the sections were sequentially immersed in anhydrous ethanol I for 5 min, anhydrous ethanol II for 5 min, anhydrous ethanol III for 5 min, xylene I for 5 min, xylene II for 5 min, and then mounted with clear neutral resin.

[0139] 5) Microscopic examination, image acquisition and analysis.

[0140] 2.3. Data Analysis and Processing

[0141] GraphPad 9.5.0 and SPSS 26 software were used for plotting and statistical analysis. Data are expressed as mean ± SEM. One-way ANOVA was used for statistical analysis to test for significant differences between the two groups. P < 0.05 was considered statistically significant.

[0142] 3. Test Results

[0143] 3.1. Effect of the formulation of this invention on the prostate coefficient in rats with benign prostatic hyperplasia.

[0144] Figure 1 The bar chart shows the prostate coefficient of rats in each group. Compared with the control group, *** p < 0.001; compared with the model group, ## p < 0.01, ### p < 0.001; compared with the Wenglitong capsule group, @@ p < 0.01, @@@ p < 0.001.

[0145] from Figure 1 It can be seen that, compared with the blank group, the prostate coefficient of rats in the model group was significantly increased; compared with the model group, the prostate coefficient of rats treated with the prescription of this invention or Wenglitong capsules was significantly reduced (P<0.01, P<0.001); and there was no significant difference in the prostate coefficient between the high-dose prescription group and the positive drug Wenglitong capsule group (P>0.05).

[0146] 3.2. Effects of the formulation of this invention on PCAP index and serum hormone index in prostate tissue of rats with benign prostatic hyperplasia.

[0147] PACP (prostatic acid phosphatase) is an enzyme secreted by prostatic epithelial cells; serum T (testosterone) is an androgen, DHT (dihydrotestosterone) is an androgen, and E2 (estradiol) is an estrogen. Elevated levels of PACP in prostatic tissue, as well as elevated levels of T, DHT, and E2 in serum, suggest possible pathological or physiological changes such as benign prostatic hyperplasia (BPH) or prostatitis.

[0148] The PCAP content in the prostate tissue of rats in each group, as well as the serum T, DHT, and E2 content, are shown in Table 1.

[0149] Table 1

[0150]

[0151]

[0152] Figure 2A Bar chart showing PCAP content in prostate tissue of rats in each group. Figure 2B The bar chart shows the serum E2 levels of rats in each group. Figure 2C The bar chart shows the serum DHT levels in rats from each group. Figure 2D The bar chart shows the serum T content of rats in each group. Compared with the control group, *** p < 0.001; compared with the model group, # p < 0.05 ## p < 0.01, ### p < 0.001; compared with the Wenglitong capsule group, @ p < 0.05 @@ p < 0.01, @@@ p < 0.001.

[0153] From Table 1 and Figures 2A-2D It can be seen that, compared with the blank group, the PACP content in the prostate tissue of rats in the model group was significantly increased (P<0.001). Compared with the model group, the PACP content in the prostate tissue of rats in the low-dose prescription group and the positive control drug Wenglitong capsule group was significantly decreased (P<0.001), the medium-dose prescription group was significantly decreased (P<0.01), and the high-dose prescription group was slightly decreased (P<0.05). Compared with the positive control drug Wenglitong capsule group, the PACP content in the prostate tissue of rats in the high-dose prescription group was slightly increased (P<0.01); there was no significant difference between the low- and medium-dose prescription groups (P>0.05).

[0154] Compared with the control group, the serum E2 level in the model group rats was significantly increased (P<0.001). Compared with the model group, the serum E2 level in the medium- and high-dose prescription groups and the positive control drug Wenglitong capsule group was significantly decreased (P<0.001), and the low-dose prescription group showed a significant decrease (P<0.01). Compared with the positive control drug Wenglitong capsule group, the serum E2 level in the low- and medium-dose prescription groups was significantly increased (P<0.001), and the high-dose prescription group showed a significant decrease (P<0.001).

[0155] Compared with the control group, the serum DHT level in the model group rats was significantly increased (P<0.001). Compared with the model group, the serum DHT level in all prescription dosage groups was significantly decreased (P<0.01), with no significant difference in the positive control group (Wenglitong capsules) (P>0.05). Compared with the positive control group (Wenglitong capsules), the serum DHT level in the high-dose prescription group was slightly decreased (P<0.05); there was no significant difference in the low- and medium-dose prescription groups (P>0.05).

[0156] Compared with the control group, the serum T content in the model group rats was significantly increased (P<0.001). Compared with the model group, the serum T content in the medium and high dose groups of the prescription was significantly decreased (P<0.001), the low dose group was significantly decreased (P<0.01), and the positive control group (Wenglitong capsules) was slightly decreased (P<0.05). Compared with the positive control group (Wenglitong capsules), the serum T content in the medium dose group was slightly decreased (P<0.05); the high dose group was significantly decreased (P<0.001), and there was no significant difference in the low dose group (P>0.05).

[0157] The above results show that low, medium, and high doses of the prescription of this invention and the positive control drug Wenglitong capsules can inhibit the levels of DHT, E2, and T in rat serum and the content of PACP in prostate tissue to varying degrees, indicating that low, medium, and high doses of the prescription of this invention and the positive control drug Wenglitong capsules all have therapeutic effects on benign prostatic hyperplasia.

[0158] 3.3. Effects of the formulation of this invention on prostate tissue in rats with benign prostatic hyperplasia.

[0159] Figure 3 HE staining images (200×) of prostate tissue from rats in each group. Among them, A is the blank group, B is the model group, C is the low-dose prescription group, D is the medium-dose prescription group, E is the high-dose prescription group, and F is the Wenglitong capsule group.

[0160] Pathological examination, such as Figure 3As shown, in the control group, a small number of samples showed acinar epithelial cell edema, a small amount of inflammatory cell infiltration in the stroma, and abundant secretions in the glandular lumen. A few samples occasionally showed epithelial cell hyperplasia. Compared with the control group, the model group showed an increased number of samples with epithelial cell hyperplasia and edema, and the edema area was larger. The number of samples with secretions in the glandular lumen was reduced, and the area of ​​edema was smaller. No obvious inflammatory cell infiltration was observed. Compared with the model group, the number of samples showing epithelial cell hyperplasia and the area of ​​epithelial cell edema were reduced in the positive control group (Wenglitong capsules), with no other significant differences. In the low-dose group, the number of samples showing epithelial cell hyperplasia and the area of ​​epithelial cell edema were reduced, while the number and area of ​​samples with visible secretions in the glandular lumen were increased, and inflammatory cell infiltration was observed in a few samples. In the medium-dose group, the number of samples showing epithelial cell hyperplasia and the area of ​​epithelial cell edema were reduced, while the number and area of ​​samples with visible secretions in the glandular lumen were increased, with no other significant differences. In the high-dose group, the number of samples showing epithelial cell hyperplasia was reduced, the number of samples with visible secretions in the glandular lumen was increased, and inflammatory cell infiltration was observed in a few samples. Compared with the positive control group (Wenglitong capsules), the number and area of ​​samples with visible secretions in the glandular lumen were increased in the low, medium, and high-dose groups; inflammatory cell infiltration was observed in a few samples in the low-dose group.

[0161] In summary, after treatment with the prescription of this invention and the positive control drug Wenglitong capsules, the prostate coefficient, PACP content in prostate tissue, and serum testosterone content in benign prostatic hyperplasia (BPH) model rats decreased. Combined with pathological results, these findings all reflect that the prescription of this invention and the positive control drug Wenglitong capsules have a therapeutic effect on BPH, and the efficacy of high-dose prescription of this invention is comparable to that of the positive control drug Wenglitong capsules.

[0162] The above description is merely a specific embodiment of this disclosure, enabling those skilled in the art to understand or implement it. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of this disclosure. Therefore, this disclosure is not to be limited to the embodiments described herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A traditional Chinese medicine composition for treating benign prostatic hyperplasia, characterized in that, The traditional Chinese medicine composition includes the following components: Seaweed, kelp, cuttlebone, Dioscorea hypoglauca, Salvia miltiorrhiza, Scrophularia ningpoensis, dandelion, Cuscuta chinensis, and Plantago asiatica.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, The traditional Chinese medicine composition comprises the following components in parts by weight: Seaweed 12-18 parts, kelp 12-18 parts, cuttlebone 18-22 parts, Dioscorea hypoglauca 28-32 parts, Salvia miltiorrhiza 18-22 parts, Scrophularia ningpoensis 28-32 parts, dandelion 22-26 parts, Cuscuta chinensis 18-22 parts, and Plantago asiatica 12-18 parts.

3. The traditional Chinese medicine composition according to claim 1 or 2, characterized in that, The traditional Chinese medicine composition comprises the following components in parts by weight: Seaweed 15 parts, kelp 15 parts, cuttlebone 20 parts, Dioscorea hypoglauca 30 parts, Salvia miltiorrhiza 20 parts, Scrophularia ningpoensis 30 parts, dandelion 24 parts, Cuscuta chinensis 20 parts, and Plantago asiatica 15 parts.

4. A pharmaceutical preparation for treating benign prostatic hyperplasia, characterized in that, The pharmaceutical preparation contains the active ingredient of the traditional Chinese medicine composition as described in any one of claims 1-3.

5. The pharmaceutical preparation according to claim 4, characterized in that, The pharmaceutical preparation also includes excipients.

6. The pharmaceutical preparation according to claim 4 or 5, characterized in that, The dosage form of the pharmaceutical preparation is granules, tablets, capsules, or decoction.

7. A method for preparing a pharmaceutical formulation as described in any one of claims 4-6, characterized in that, The preparation method includes the following steps: (1) The traditional Chinese medicine composition according to any one of claims 1-3 is subjected to water extraction to obtain an aqueous extract; (2) The aqueous extract is prepared into the dosage form of the drug preparation.

8. The preparation method according to claim 7, characterized in that, The water extraction step includes: The herbal composition is decocted with water at least twice, each time for 1-4 hours. After each decoction, the mixture is filtered, and the filtrates are combined to obtain the aqueous extract.

9. The preparation method according to claim 7 or 8, characterized in that, In the water extraction step, the total mass of water is 18-35 times the mass of the traditional Chinese medicine composition.

10. The preparation method according to claim 8, characterized in that, In the water extraction step, the decoction is performed twice. In the first decoction, the mass of water is 10-20 times the mass of the Chinese herbal composition, and the decoction time is 2-3 hours. In the second decoction, the mass of water is 8-15 times the mass of the Chinese herbal composition, and the decoction time is 1-2 hours.

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