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

Abstract

The invention discloses a traditional Chinese medicine composition for preventing and treating benign prostatic hyperplasia and a preparation method thereof. The traditional Chinese medicine composition comprises 30-70 parts of pollen typhae and 10-30 parts of talc. The traditional Chinese medicine composition can obviously inhibit the volume increase of prostate, improve tissue lesion of prostatic hyperplasia, inhibit prostatic cell proliferation mediated by androgen receptors and improve inflammation symptoms of benign prostatic hyperplasia glands, thereby being used for treating benign prostatic hyperplasia.

Description

Technical Field

[0001] The invention relates to a traditional Chinese medicine composition for preventing and treating benign prostatic hyperplasia and a preparation method thereof, belonging to the technical field of pharmaceuticals. Background Art

[0002] Benign prostatic hyperplasia (BPH) is a common disease of the urinary system in middle-aged and elderly men and is closely related to age. Epidemiological studies show that 50% of men over 60 years old can observe BPH symptoms at the tissue level; the prevalence of BPH in men over 80 years old is close to 90%. The prostate is half-surrounded by the bladder neck and urethra, and is a solid glandular organ in the shape of an inverted chestnut. When it proliferates to a certain extent, it compresses the bladder outlet urethra and causes it to deform, resulting in increased urethral resistance and difficulty in urine discharge.

[0003] The cause of prostatic hyperplasia is complex, and there are many theories. The hormone theory mainly focuses on the androgen receptor (AR) pathway. Testosterone is reduced to dihydrotestosterone under the action of 5α reductase. Dihydrotestosterone has a higher binding affinity with the androgen receptor. Excessive binding of dihydrotestosterone to the androgen receptor leads to AR-mediated excessive proliferation of prostate cells, which in turn causes benign prostatic hyperplasia. In addition, chronic inflammation is another major cause of prostatic hyperplasia. Proinflammatory stimulators promote the growth of prostatic epithelial cells, and in clinical studies, prostatic hyperplasia tissue contains inflammatory infiltration. In summary, AR-mediated cell proliferation and inflammation are the main causes of BPH.

[0004] BPH patients are generally older. Considering the physical fitness of the affected population and many postoperative complications, drug therapy is the main treatment for BPH. Currently, common clinical drugs for the treatment of BPH include 5α-reductase inhibitors, α-receptor blockers, etc., which have significant therapeutic effects in inhibiting the increase of prostate volume, but have certain side effects. For example, α1 adrenergic receptor blockers can cause dizziness, weakness, orthostatic hypotension and abnormal ejaculation in patients. 5α-reductase inhibitors may cause non-alcoholic fatty liver disease, insulin resistance, type 2 diabetes, dry eyes, potential renal dysfunction and other metabolic dysfunctions. With the deepening of the aging population in my country, the incidence of BPH in my country has also increased year by year, so there is an urgent need to find a better drug to treat benign prostatic hyperplasia. Summary of the invention

[0005] The purpose of the present application is to provide a Chinese medicine composition for preventing and treating benign prostatic hyperplasia, and a preparation method and application thereof.

[0006] The purpose of the present invention can be achieved by the following technical solutions:

[0007] In a first aspect, the present invention claims protection for a traditional Chinese medicine composition for resisting prostatic hyperplasia, which comprises 30-70 parts of Pollen Typhae and 10-30 parts of Talc by weight.

[0008] Furthermore, the Chinese medicine composition comprises 30-60 parts of Pollen Typhae and 10-30 parts of Talc. Still further, the Chinese medicine composition comprises 42 parts of Pollen Typhae and 18 parts of Talc.

[0009] Furthermore, the preparation method of the Chinese medicine composition comprises the following steps:

[0010] After passing Typhae pollen and talc through a 100-mesh sieve, mix them in proportion, add 8-12 times the volume of 95% (v / v) ethanol, soak for 0.8-1.2 hours, heat and reflux for extraction for 0.8-1.2 hours, and collect the supernatant filtrate;

[0011] Then add 6-10 times the volume of 50% (v / v) ethanol to the residue, heat and reflux to extract for 0.8-1.2h, and collect the supernatant filtrate;

[0012] Then add 6-10 times the volume of ultrapure water to the filter residue, heat and reflux for extraction for 0.8-1.2h, and collect the supernatant filtrate;

[0013] The filtrates obtained from the three times were combined and concentrated under reduced pressure to obtain a clear paste. The filtrate was concentrated under reduced pressure at -0.08MPa and 60°C to a clear paste with a relative density of 110-120 at 55°C, and dried in a vacuum drying oven at 60°C to obtain a dry paste powder.

[0014] In a second aspect, the present invention seeks to protect a drug for preventing prostatic hyperplasia, which is prepared into a pharmaceutically acceptable dosage form by the above-mentioned Chinese medicine composition and a pharmaceutically acceptable carrier and / or excipient.

[0015] Furthermore, the pharmaceutically acceptable carrier or excipient is at least one of an excipient, a solubilizer, a preservative, a stabilizer, a wetting agent, an emulsifier, a salt for regulating osmotic pressure, and a buffer; the pharmaceutically acceptable dosage form is at least one of a capsule, a granule, a tablet, an oral solution, a pill, a soft capsule, a honey pill, a water pill, a powder injection, an injection solution, a spray, and a dispersion.

[0016] In a third aspect, the present invention seeks to protect the use of the above-mentioned traditional Chinese medicine composition in the preparation of a drug for treating prostate hyperplasia.

[0017] Beneficial effects of the present invention:

[0018] The present invention establishes a benign prostatic hyperplasia model by inducing testosterone propionate, and adopts histological and molecular biological methods to find that the Chinese medicine composition can significantly reduce the wet weight of the prostate, reduce the prostate index and volume; morphologically, it is observed that prostatic hyperplasia is significantly reduced; and the PSA level in serum is significantly reduced. It is fully verified that the Chinese medicine mixture has a significant improvement effect on BPH, and the composition can be used to prepare or prevent prostatic hyperplasia drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. It is more obvious that the following description is only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0020] Figure 1 This is a rat model of prostatic hyperplasia, and the prostate morphology of rats in each group after administration.

[0021] Figure 2 This is a rat model of prostate hyperplasia, and the prostate index results of rats in each group after drug administration.

[0022] Figure 3 This is a rat model of prostate hyperplasia, and the prostate volume results of rats in each group after administration.

[0023] Figure 4 This is a rat model of prostatic hyperplasia. The PSA content in the serum of rats in each group after administration was measured.

[0024] Figure 5 This is a rat model of prostate hyperplasia. The HE staining results of the prostate tissue of each group of rats after drug administration. The upper picture in each figure is the result under a 50x microscope, marked as 50×, and the lower picture is the result under a 200x microscope, marked as 200×.

[0025] Figure 6 This is a rat model of prostate hyperplasia. The results of the expression of androgen receptor AR protein in the prostate tissue of each group of rats after drug administration. The picture in the figure is the result under a 50x microscope, marked as 50×.

[0026] Figure 7 This is a rat model of prostate hyperplasia. The results of the expression of proliferating cell nuclear antigen PCNA protein in the prostate tissue of each group of rats after drug administration. The pictures in the figure are the results under a 50x microscope, marked as 50×.

[0027] Figure 8 This is a rat model of prostate hyperplasia. The results of the expression of interleukin-6 (IL-6) protein in the prostate tissue of each group of rats after drug administration. The pictures in the figure are the results under a 50x microscope and are marked as 50×.

[0028] Fig. 9 The rat model of prostatic hyperplasia was established. The TNF-α content in the serum of rats in each group after administration was changed. DETAILED DESCRIPTION

[0029] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention is described clearly and completely below. Obviously, the described embodiments are part of the embodiments of the present invention, but not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work belong to the scope of protection of the present invention.

[0030] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0031] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as commonly understood by those skilled in the art. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the contents of this specification shall prevail.

[0032] The purpose of the present invention is to provide an application of Pollen Typhae in the preparation of a medicine for preventing or treating prostatic hyperplasia, which has excellent pharmacological and pharmacodynamic effects.

[0033] Unless otherwise specified, the reagents and materials used in the following examples can be obtained through commercial channels.

[0034] Example 1

[0035] Composition 1: Puhuang 42g, Talc 18g.

[0036] Composition 2: 30g of Puhuang and 30g of Talc.

[0037] Composition three: 50g of Puhuang and 10g of talc.

[0038] The preparation process is as follows:

[0039] The above two herbs, pollen and talcum powder are passed through a 100-mesh sieve, mixed and set aside; add 95% ethanol with a volume of 10 times the water and soak for 1 hour. Heat and reflux for extraction for 1 hour, and collect the supernatant filtrate. Then add 500mL of 50% ethanol to the filter residue, heat and reflux for extraction for 1 hour, and collect the supernatant filtrate. Then add 500mL of ultrapure water to the filter residue, heat and reflux for extraction for 1 hour, and collect the supernatant filtrate. Combine the filtrates obtained three times, and concentrate under reduced pressure to obtain a clear paste. When the filtrate is concentrated under reduced pressure to 55°C at -0.08MPa and 60°C, the relative density of the clear paste is measured to be 110-120, and it is placed in a vacuum drying oven at 60°C to dry to obtain dry paste powder.

[0040] Preparation of granules: Mix the dry paste powder, sucrose and dextrin, granulate, dry and granulate, make 1000g granules according to conventional process, and bag them to obtain granules.

[0041] Preparation of capsules: Mix the dry paste powder, sucrose and dextrin, granulate, dry, granulate and encapsulate. According to conventional process, 1000 granules are made and packed into bags to obtain capsules.

[0042] Preparation of tablets: Mix the dry paste powder, sucrose and dextrin, granulate, dry, granulate and tablet, and make 1000 tablets according to conventional process to obtain tablets.

[0043] Embodiment 2:

[0044] 1. Methods and Materials

[0045] 1.1 Animal selection

[0046] 104 male SD rats (180-200 g, Shanghai Institute of Biomedical Technology Laboratory Animal Management Department, license number: SCXK (Shanghai) 2018-0006) were used. They were given humane care in accordance with the 3R principles of experimental animal use.

[0047] 1.2 Drugs and reagents

[0048] Testosterone propionate (purity: ≥98%, McLean, batch number: C13133860), soybean oil (reagent grade, McLean, batch number: C13043318), finasteride (McLean, batch number: C12005059), sodium carboxymethyl cellulose (CMC-Na, Sinopharm Chemical Reagent Co., Ltd., batch number: 20130314), Fina (reagent grade, McLean, batch number: 20230611 Chuntang Chinese Medicine Pieces Co., Ltd., batch number: 220901), prostate specific antigen (PSA) kit, tumor necrosis factor TNF-α kit.

[0049] 1.3 Instruments

[0050] TB-25 1 / 100,000 electronic balance (Denver Instrument (Beijing) Co., Ltd.); P-214 1 / 10,000 balance (Denver Instrument (Beijing) Co., Ltd.), GZX-9070MBE electric heating blast drying oven (Shanghai Boxun Industrial Co., Ltd. Medical Equipment Factory), ultrafine pulverizer (Shandong Jinan Tianyu Special Equipment Co., Ltd.).

[0051] 1.4 Experimental methods

[0052] 1.4.1 Grouping

[0053] Forty male clean-grade SD rats were randomly divided into five groups: negative control group, model group, positive drug group (finasteride 1 mg kg -1 ), Puhuisan drug original powder (dry paste powder prepared according to the formula of composition 1 in Example 1) low and high groups. During the process, the room temperature was controlled at 20℃~25℃, the humidity was 40%~70%, the light was 12h / d, and the animals were allowed to eat and drink freely. The experiment started after one week of adaptation to the environment.

[0054] 1.4.2 Modeling and drug administration methods

[0055] 40 normal male rats were fed adaptively for 1 week, 8 rats were randomly selected as blank control group, and the remaining 32 rats were used to establish the model of benign prostatic hyperplasia. The model group was subcutaneously injected with testosterone propionate injection 5 mg / kg·d at the back of the neck, and the blank control group was injected with an equal volume of soybean oil for 28 consecutive days and fed normally to establish the rat model of benign prostatic hyperplasia.

[0056] In this embodiment, the low dose dosage is 2.205g / kg / d, which is the clinical equivalent dosage; the high dose dosage is 4.21g / kg / d, which is twice the clinical equivalent dosage. In this application, the clinical equivalent dosage refers to the whole prescription raw drug amount of 10g (including 7g of pollen and 3g of talc) for adults (calculated as 60kg) per day, the human-mouse coefficient is 6.3, and the dosage based on rats is 2.205g / kg / d.

[0057] After 28 days of modeling, drug administration began. The blank group remained unchanged, while the negative control group, model group, low-dose drug group and high-dose drug group continued to be administered and modeled for 28 days.

[0058] Determination of water-loaded urine volume in BPH model rats: In the 1st to 4th week after the start of drug administration, after intragastric administration of the test drug and subcutaneous injection of testosterone propionate injection, each rat was intragastrically administered with 30 mL kg of normal saline, and immediately placed in a metabolic cage with a disposable plastic cup at the bottom. The metabolic urine at the 6th hour was collected and weighed, and this was done once a week for 4 consecutive weeks.

[0059] 1.4.3 Specimen collection and processing

[0060] 24 hours after the last administration, the body weight of rats in each group was weighed and recorded. Rats were anesthetized by intraperitoneal injection of 20% (g / 100ml) urethane, the abdominal cavity was opened, blood was collected through the abdominal aorta, and the blood was left to stand at room temperature for 1h. The serum was separated by low-temperature centrifugation (4℃, 3500r·min-1, 10min) and stored at -80℃. The prostate tissue of both sides was removed, and the surrounding fascia and adipose tissue were carefully removed. The prostate was rinsed in pre-cooled saline, and the weight of the prostate was weighed by an electronic balance after being wiped dry with filter paper. The prostate index (PI) was calculated (PI = prostate wet weight (g) / body weight (g) × 100). The dorsolateral lobe of the prostate was fixed in 4% paraformaldehyde solution for tissue morphology detection. The mesentery and fat around the urogenital system were carefully removed with surgical instruments, and then the epididymis was separated and rinsed with 1×PBS. Cut the epididymis into several sections, squeeze it with tweezers, squeeze the sperm into saline, and incubate it in a cell culture incubator for 20 minutes to allow the sperm to disperse on their own. Take out the culture dish and add saline to dilute the sperm suspension 100 times, then use a pipette to absorb the suspension and drop it into the gap between the counting platform of the hemocytometer and the edge of the cover glass, so that the suspension penetrates and fills the counting chamber, and the number of sperm is measured under an optical microscope.

[0061] 1.4.4 HE staining pathological detection

[0062] The dorsolateral lobe of the rat prostate was fixed in 4% paraformaldehyde solution (commercially available) for 48 hours, rinsed with running water for 3 hours, dehydrated in ethanol gradient, transparentized with xylene, embedded in conventional paraffin, cooled at room temperature overnight, blocked and sliced, and 4-micron pathological sections were made. The sections were stained with hematoxylin-eosin and sealed with Zhongxing numerical sealant, and the morphological changes of prostate tissue were observed under an optical microscope.

[0063] 1.4.5 Immunohistochemistry and pathological testing

[0064] Prostate tissue sections of rats were taken for immunohistochemical staining to detect the expression of AR protein, PCNA protein and IL-6 protein in the epithelial and interstitial regions of prostate tissue. Under optical microscopy, the cell nucleus was blue, and the protein positive expression was brown-yellow. Five discontinuous high-power microscopic fields (50×) were taken, and the average optical density value (IOD / Area) of positive staining in each field was calculated.

[0065] 1.4.6 Serum A, TNF-α detection

[0066] Take rat serum frozen in -80℃ refrigerator and measure it strictly according to the operating instructions of PSA Elisa detection kit.

[0067] 1.4.7 Statistical methods

[0068] GraphPad Prism 9.0 software was used for statistical analysis and the data were expressed as Mean±SD. One-way ANOVA was performed among the groups, and p<0.05 was considered statistically significant.

[0069] 2. Results

[0070] 2.1 Effect of Puhuisan on the prostate of BPH rats

[0071] according to Figure 1 and Figure 3 It can be seen that compared with the blank group, the prostate volume of the model group rats increased significantly, and testosterone propionate can significantly induce prostate enlargement; after administration, the prostate volume of the low-dose group and the high-dose group was significantly reduced compared with the model group, and the volume of the high-dose group rats was close to that of the sham operation group. Figure 2 The results showed that the prostate index of the model group rats increased significantly compared with the blank group (n=8, p # <0.01); after administration, the low-dose group and the high-dose group were significantly lower than the model group (n=8, p ** <0.01). This indicates that Puhuisan can significantly inhibit the increase of prostate volume in rats with benign prostatic hyperplasia and significantly reduce the prostate index.

[0072] 2.2 Pathological analysis of prostate staining of BPH rats with Puhuisan

[0073] according to Figure 5 The results of HE staining pathological analysis showed that the prostate glandular epithelial cells in the model group were arranged neatly; the glandular epithelium in the model group was thickened and disorderly arranged, and prostate hyperplasia occurred; the disordered conformation of the glandular epithelial cells in the low-dose and high-dose groups was improved to varying degrees, indicating that the Puhuisan of the present application can improve the tissue lesions of prostate hyperplasia.

[0074] 2.3 Effect of Puhuisan on serum PSA level in BPH rats

[0075] according to Figure 4 It can be seen that compared with the blank group, the serum PSA content in the model group was significantly increased; the low-dose group and the high-dose group significantly inhibited this increase. From the perspective of clinical detection, it is explained that Puhuisan can reduce the increase in serum PSA content caused by prostate hyperplasia.

[0076] 2.4 Effect of Puhuisan on urine output in BPH rats

[0077] According to Table 1, compared with the blank group, the urine output of the model group was significantly reduced, indicating that prostate hyperplasia can cause symptoms of dysuria and reduced urine output; after four weeks of administration of low-dose and high-dose drugs, the urine output of rats increased significantly, indicating that Puhuisan can increase urine output and improve the symptoms of prostate hyperplasia.

[0078] Table 1 Urine output of rats by water loading method after 1-4 weeks of drug administration (n=8)

[0079]

[0080] 2.5 Effect of Puhuisan on the expression of AR and PCNA in prostate tissue of rats

[0081] according to Figure 6 and Figure 7 It can be seen that compared with the blank group, the expression levels of AR and PCNA proteins in the model group were significantly increased, indicating that prostate hyperplasia will cause increased expression of androgen receptors, thereby promoting the increased expression of proliferating cell nuclear antigen PCNA; the low-dose group and the high-dose group significantly inhibited this increase, indicating that Puhuisan treats benign prostate hyperplasia by mediating AR-mediated prostate cell proliferation.

[0082] 2.6 Effect of Puhuisan on IL-6 expression in prostate tissue and TNF-α content in serum of rats.

[0083] according to Figure 8 and Fig. 9 It can be seen that compared with the blank group, the IL-6 protein expression and TNF-α content in serum of the model group were significantly increased, indicating that prostatic hyperplasia can cause inflammation of prostatic tissue, thereby promoting the development of prostatic hyperplasia; the low-dose group and the high-dose group significantly alleviated inflammation, indicating that Puhuisan treats benign prostatic hyperplasia by exerting anti-inflammatory effects.

[0084] From the above results, it can be seen that Puhuisan can significantly inhibit the increase of prostate volume, improve the tissue lesions of prostate hyperplasia, and improve the symptoms of benign prostatic hyperplasia. Therefore, it can be used to treat and / or prevent benign prostatic hyperplasia, and thus used to prepare drugs for treating and / or preventing benign prostatic hyperplasia.

[0085] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should fall within the protection scope determined by the claims of the present invention.

Claims

1. A Chinese medicine composition for resisting prostatic hyperplasia, characterized in that: The traditional Chinese medicine composition comprises 30-70 parts of pollen and 10-30 parts of talc by weight.

2. The Chinese medicine composition according to claim 1, characterized in that: The traditional Chinese medicine composition comprises 30-60 parts of pollen pollen and 10-30 parts of talc.

3. The Chinese medicine composition according to claim 2, characterized in that: The traditional Chinese medicine composition comprises 42 parts of pollen typhae and 18 parts of talc.

4. The Chinese medicine composition according to claim 1, 2 or 3, characterized in that: The preparation method of the Chinese medicine composition comprises the following steps: After passing Typhae pollen and talc through a 100-mesh sieve, mix them in proportion, add 8-12 times the volume of 95% ethanol, soak for 0.8-1.2 hours, heat under reflux for extraction for 0.8-1.2 hours, and collect the supernatant filtrate; Then add 6-10 times the volume of 50% ethanol to the filter residue, heat and reflux to extract for 0.8-1.2h, and collect the supernatant filtrate; Then add 6-10 times the volume of ultrapure water to the filter residue, heat and reflux for extraction for 0.8-1.2h, and collect the supernatant filtrate; The filtrates obtained from the three times were combined and concentrated under reduced pressure to obtain a clear paste. The filtrate was concentrated under reduced pressure at -0.08MPa and 60°C to a clear paste with a relative density of 110-120 at 55°C, and dried in a vacuum drying oven at 60°C to obtain a dry paste powder.

5. A drug for preventing prostatic hyperplasia, characterized in that: A pharmaceutically acceptable dosage form is prepared from the Chinese medicine composition according to any one of claims 1 to 4 and a pharmaceutically acceptable carrier and / or excipient.

6. The drug for treating prostatic hyperplasia according to claim 5, characterized in that: The pharmaceutically acceptable carrier or excipient is at least one of an excipient, a solubilizer, a preservative, a stabilizer, a wetting agent, an emulsifier, a salt for adjusting osmotic pressure, and a buffer; the pharmaceutically acceptable dosage form is at least one of a capsule, a granule, a tablet, an oral solution, a pellet, a soft capsule, a honey pill, a water pill, a powder injection, an injection, a spray, and a dispersion.

7. Use of the Chinese medicine composition according to any one of claims 1 to 4 in the preparation of a drug for treating prostatic hyperplasia.

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