CN1846775A - New pharmaceutical application of traditional Chinese medicine nutmeg and its extract - Google Patents

Abstract

The invention relates to the new application of the traditional Chinese medicine Nutmeg and its extract in the field of pharmacy, in particular to the application of the traditional Chinese medicine Nutmeg and its extract in the preparation of 5α-reductase inhibitors and drugs for the treatment of benign prostatic hyperplasia. Low cost and easy to use.

Description

New pharmaceutical application of traditional Chinese medicine nutmeg and its extract

technical field

The invention relates to a new application of the traditional Chinese medicine nutmeg, in particular to the application in the preparation of 5α-reductase inhibitors and drugs for treating benign prostatic hyperplasia, and belongs to the field of traditional Chinese medicines.

Background technique

Benign prostatic hyperplasia (BPH), namely prostatic hypertrophy, is a common disease in elderly men. It is characterized by static obstruction caused by benign prostatic hyperplasia and dynamic obstruction caused by increased prostate smooth muscle tension. The clinical manifestations are bladder urine outflow obstruction, Urinary urgency, frequent urination, and residual urine often lead to urinary tract infection and even induce nephritis. The incidence increases with age, and the number of incidences has also increased significantly in recent years, seriously affecting patients' physical activities and social skills, thereby affecting their quality of life. BPH mostly adopts surgery to treat well, but surgery is not without risk for elderly patients. BPH surgery is a traumatic and destructive operation with a risk of death. Therefore, drug treatment is sought to avoid the risk of surgery, and, with With the continuous improvement of the understanding of the pathogenesis of benign prostatic hyperplasia, drugs have become an important means of BPH treatment.

Although the exact mechanism of BPH has not been fully elucidated, the relationship between 5-α reductase and BHP has been widely accepted. The role of 5-alpha reductase is to metabolize testosterone (T) into dihydrotestosterone (DHT), which is related to the occurrence of BPH. At the same time, studies have shown that: 5α-reductase is a key enzyme in skin androgen metabolism. In recent years, it has been found that various skin diseases are related to abnormal androgen metabolism. Some 5α-reductase inhibitors have been used for androgenetic alopecia and other androgen Related skin diseases, such as acne, hirsutism, etc. (Yin Xingping, Xia Longqing. The role of 5a-reductase in androgen metabolism and the application of its inhibitors in dermatology. Foreign Medicine Dermatology, 2004, 30( 6): 375-377). DHT is the active form of prostate androgen, which is necessary for the differentiation, growth and development of the prostate. However, in old age, the prostate tissue maintains a high 5-α reductase activity, which increases the DHT content, so it is considered to be an important factor leading to benign prostatic hyperplasia. This theory has been confirmed by the study of finasteride, a 5-alpha reductase inhibitor. Finasteride competitively combines with 5α-reductase to reduce the combination of testosterone and 5α-reductase, blocks the conversion of testosterone to DHT, rapidly reduces plasma DHT and DHT in the prostate, and finally shrinks the prostate and increases the urinary flow rate. Improve the symptoms of obstruction and reduce the complications of BPH. In recent years, plant-derived drugs have shown good prospects for the treatment of BPH, such as blue palm fruit extract and pollen extract (Shenitong), both of which have the effect of inhibiting 5α-reductase.

Nutmeg is the mature seed kernel of Myristica fragrans Houtt, a tall evergreen tree plant of Myristicaceae. It is native to the Maluku Islands and introduced and cultivated in Taiwan, Yunnan and Guangdong. According to the "Materia Medica" records: "The pungent taste of nutmeg can dissipate and disappear, and the temperature can be unobstructed. Its air is fragrant, and the aroma first enters the spleen, and the spleen governs digestion. It is mild and spicy, so it appetizers, and the stomach likes warmth." "Chinese Materia Medica" believes that: Nutmeg is pungent, warms the spleen, stomach, and large intestine meridian, has the effects of astringent intestines, relieving diarrhea, and warming the middle and promoting qi. It shows that nutmeg contains tannic acid components of volatile oil, fatty oil, starch, protein and gallic acid. The volatile oil contains myristyl ether, safrole, elemene, methylcresol and various terpene compounds. Its main Pharmacological effects are: antimicrobial, anti-oxidation, anesthesia, sedation and hallucinogenic effects ("Chinese Materia Medica", edited by Zhang Tingmo, Beijing, Higher Education Press, 2002, 1). But it has not been seen that nutmeg inhibits 5α-reductase, Reports on the prevention or treatment of benign prostatic hyperplasia and related diseases.

Contents of the invention

The technical solution of the invention is to provide a new application of the traditional Chinese medicine nutmeg and its extract. The invention also provides a pharmaceutical composition containing the original drug or extract of nutmeg as an active ingredient, a preparation method and application thereof.

The invention provides the application of the traditional Chinese medicine nutmeg in the preparation of 5α-reductase inhibitor medicine and the application of the traditional Chinese medicine nutmeg in the preparation of the medicine for treating benign prostatic hyperplasia.

The traditional Chinese medicine nutmeg comes from the mature kernel of Myristica fragrans Houtt, a tall evergreen tree plant of Myristicaceae.

The nutmeg is raw medicinal powder of nutmeg or water/organic solvent extract.

The present invention also provides a pharmaceutical composition for the preparation of 5α-reductase inhibitor drugs, which is composed of the raw nutmeg powder or its water/organic solvent extract as the active ingredient, plus pharmaceutically acceptable Preparations prepared from excipients or auxiliary ingredients.

Wherein, the nutmeg organic solvent extract is one of organic solvents such as methanol, ethanol, ethyl acetate, n-butanol, ether, petroleum ether, or a mixed solvent composed of any two or more solvents in any proportion. of extracts.

After the above-mentioned traditional Chinese medicine nutmeg or its extract is mixed with pharmaceutically acceptable auxiliary additive components, the 5α-reductase inhibitor, the medicine for preventing and/or treating benign prostatic hyperplasia can be prepared according to the corresponding conventional pharmaceutical preparation method. For example, after mixing with commonly used auxiliary ingredients such as disintegrants, excipients, lubricants, binders, and fillers that are acceptable in oral preparations, according to conventional operating methods and processes, it can be made into tablets, Drugs in the form of pills, capsules or various corresponding sustained-release agents, controlled-release agents and other solid oral preparations; with conventional solubilizers, emulsifiers, modifiers, foaming or antifoaming agents and other surfactants, diluents, After mixing preservatives, stabilizers, flavoring agents, thickeners, etc., according to the corresponding conventional methods, it can be prepared into oral medicines in the form of liquid preparations such as liquid preparations, syrups, and oral liquids; with conventional suspending agents, stabilizers, The dispersing agent or the reagent for adjusting the osmotic pressure of the solution are mixed, and the injection is prepared according to the corresponding conventional method.

Further, the present invention provides a preparation method of the pharmaceutical composition, wherein the preparation of the nutmeg extract comprises the following steps:

a) Nutmeg was extracted 1-3 times with water or 30-95% ethanol under reflux, the amount of solvent was 6-8 times respectively, the extraction time was 1-3 hours, the extracts were combined, concentrated and dried to obtain sample A;

b) A is dissolved in water, extracted with petroleum ether, diethyl ether, ethyl acetate or n-butanol, and the solvent is recovered under reduced pressure to obtain a water extract and a corresponding organic solvent extract.

The invention also provides the application of the pharmaceutical composition in the preparation of 5α-reductase inhibitor drugs.

The invention also provides the application of the pharmaceutical composition in the preparation of drugs for treating benign prostatic hyperplasia.

According to the present invention, the above-mentioned compound preparation containing the Chinese medicine Nutmeg or its extract, or the Chinese medicine Nutmeg and its extract and other active ingredients of Western medicine or Chinese medicine which have the effect of preventing and/or treating prostate hyperplasia can be formulated according to the conventional method. Preparation technology, prepared into oral preparations or injections.

The invention extracts the nutmeg extract from the traditional Chinese medicine nutmeg, and tests prove that it has the effect of inhibiting the activity of 5α-reductase, and has obvious therapeutic effect on benign prostatic hyperplasia in male rats. It can be used for preparing 5α-reductase inhibitors, treating and/or preventing pharmaceutical preparations for benign prostatic hyperplasia, such as oral preparations or injections. It has the characteristics of remarkable curative effect, low price and convenient taking.

Apparently, according to the above content of the present invention, according to the ordinary technology and conventional means in this field, without departing from the above basic technical idea of the present invention, other various forms of modification, replacement or change can also be made.

The above-mentioned content of the present invention will be further described in detail below through specific implementation in the form of examples. However, it should not be understood that the scope of the above subject matter of the present invention is limited to the following examples, and all technologies realized based on the above content of the present invention belong to the scope of the present invention.

Detailed ways

The preparation of the capsule of the original powder of nutmeg of embodiment 1

Take the dried and mature seed kernels of the traditional Chinese medicine Myristica fragrans Houtt, grind them according to conventional methods, and sieve to make nutmeg fine powder with a particle size of 60-80 meshes. The fine powder is packed into capsules, sterilized, and packed separately.

The preparation of embodiment 2 nutmeg extract

Weigh 1000g of nutmeg, extract three times with 90% ethanol under reflux, the amount of solvent is 8 times, 6 times, 6 times respectively, and the extraction time is 2 hours, 1 hour, 1 hour respectively. Combine the extracts and recover until there is no alcohol smell. Dry under reduced pressure at 50°C to obtain sample A. Take sample A 50g, dissolve with 150ml distilled water. The aqueous solution was extracted with 3 times the volume of petroleum ether, ether, ethyl acetate, and n-butanol, and the solvent was recovered under reduced pressure to obtain samples B (petroleum ether phase), C (ether phase), D (ethyl acetate phase), and E ( Butanol phase), F (water phase).

The preparation of embodiment 3 tablet

Composition of the tablet: Nutmeg Extract A 0.5g

HPMC _LV100 30g

                                         

Magnesium stearate 0.5g

Total 100g

Mix nutmeg extract A, HPMC _LV100 , and lactose, use 75% ethanol as a binder to make wet granules, pass through a 22-mesh sieve, dry at 50°C for 3 hours, granulate with a 22-mesh sieve, add magnesium stearate and mix well Tablet, made into 1000 pieces, each piece 0.1g. Take orally, 3 times a day, 2 tablets each time.

The preparation of embodiment 4 capsules

Capsule composition: nutmeg extract B 0.5g

                               

Magnesium stearate 0.5g

Total 100g

Mix nutmeg extract B, starch and magnesium stearate evenly, pack into capsules, 1000 capsules in total, 0.1g per capsule. Take orally, 3 times a day, 2 capsules each time.

The preparation of embodiment 5 injection

Injection composition: Nutmeg extract C 0.5g

Tween 80 10ml

Sodium Chloride 8g

Nutmeg extract C, add 10% sodium carbonate to adjust the pH value to 7.0-7.5, refrigerate and filter, add Tween 80, add water for injection to 1000ml, filter, sub-package, potting, steam sterilization at 100°C, 30min Instantly.

The preparation of embodiment 6 compound tablet or capsule

Prepare the compound preparation with nutmeg extract, take 0.5g of nutmeg extract D, 200g of turmeric extract, add auxiliary materials according to the conventional method, and make tablets or capsules. 200mg per tablet or capsule, for patients, orally, 3 times a day, 1 tablet or 1 capsule each time, one month as a course of treatment.

Prove beneficial effect of the present invention by pharmacodynamic test below.

Test Example 1 Inhibitory effect of nutmeg extract on prostate cancer cell 5α-reductase activity

Prostate cancer cells (DU145, expressing 5α-reductase) were maintained and cultured in 1640 medium containing 10% calf serum, 100ug/ml streptomycin, 100U/ml penicillin according to conventional methods, and the next day, 2×10 ⁴ / The wells were inoculated in a 24-well plate, cultivated for 24 hours at 37° C. in a 5% CO _saturated humidity incubator, changed fresh medium, added 0.1 mM testosterone to each well and added samples to be tested respectively, and used blue palm fruit (Sabalberry extract, Germany Dr. Ma Pharmaceutical Factory) is the positive control drug, continue to cultivate for 2 hours, collect the cell culture supernatant, quantitatively detect dihydrotestosterone with an EIA kit, measure the absorbance value under the 450nm condition, calculate the dihydrotestosterone amount according to the standard measured value, In order to determine the conversion of testosterone and the inhibition of the sample to the conversion of testosterone, the results are shown in Table 1:

Table 1 Nutmeg inhibits the conversion of testosterone in prostate cancer cells (sample concentration: 100 μg/ml) sample name DHT (pg/ml) sample name DHT (pg/ml) Cell control cells + testosterone blue palm fruit AB 71987.38276.03431.93383.27 CDEF 179.32464.29449.31479.45

The results in Table 1 show that the DHT concentration of the cell+testosterone group was significantly higher than that of the cell control group, and the DHT concentration of each drug group was significantly lower than that of the cell+testosterone group, indicating that the nutmeg extracts A, B, C, D, E, and F were all It has the effect of inhibiting the activity of 5α-reductase in prostate cancer cells, among which nutmeg extract C has the strongest inhibitory effect on 5α-reductase, and the effect is stronger than that of the control drug blue palm fruit.

Test Example 2 Inhibitory effect of nutmeg extract on rat prostate tissue 5α-reductase activity

Take the rat prostate tissue, add homogenization buffer (1:5 volume), the composition of the homogenization buffer is: 0.88M sucrose, 1.5mM CaCl ₂ , the tissue homogenate is centrifuged at high speed (10000×g, 15min, 4°C ), take the supernatant and react with the test sample or positive drug (finasteride: produced by Merck Sharp & Dohme (UK) Ltd, specification: 5mg×10 grains/box, batch number: 235820) for 10 minutes, add 0.05mM testosterone, 0.02mM NADPH, measured the absorbance value at 450nm, and calculated the inhibition rate: inhibition rate (%)=(A ₀ -A ₁ )/A ₀ ×100, the results are shown in Table 2:

Table 2 Nutmeg inhibits the activity of prostate tissue 5α-reductase (sample concentration: 100 μg/ml) sample name Inhibition rate(%) sample name Inhibition rate(%) Cell Control Homogenate + Testosterone Finasteride AB --1006780.75 CDEF 93.7263.2469.3169.45

The results in Table 2 show that each drug group has an inhibitory effect on the activity of 5α-reductase in prostate tissue, indicating that nutmeg extracts A, B, C, D, E, and F all have the effect of inhibiting the activity of 5α-reductase in prostate tissue , wherein the nutmeg extract C has the strongest inhibitory effect on 5α-reductase, the inhibition rate reaches 93.72%, and the effect is slightly weaker than that of the control drug finasteride (100% inhibition).

Experimental example 3 Nutmeg extract C treats the effect of benign prostatic hyperplasia in rats

56 normal male SD rats were selected and randomly divided into seven groups, 8 rats in each group, i.e. normal group (gavage with equal volume of normal saline), castration group (removal of both testicles), castration+testosterone group (removal of Bilateral testes, inject testosterone propionate 2mg/kg), nutmeg high, medium and low dose groups (castration + testosterone and intragastric administration of nutmeg C extract), positive drug group (castration + testosterone and intragastric administration Sernitone (10 tablets × 1 board/box, each tablet contains water-soluble pollen extract P5 70mg, fat-soluble pollen extract EA10 4mg, produced by Nanjing Meirui Pharmaceutical Co., Ltd.)). After 45 days of administration, blood was collected from the retro-orbital venous plexus, and the serum was separated to determine the concentration of DHT. The detection method and calculation were carried out according to the instructions of the EIA kit; After weighting, place the homogenate at low temperature, centrifuge to collect the supernatant, and detect the DHT and T concentrations in the tissue. The detection method and calculation are carried out according to the instructions of the EIA kit. The results are shown in Table 3.

Table 3 Nutmeg extract C on the serum and tissue DHT levels of benign prostatic hyperplasia rats (n=8, X ± SD) group Dose mg/kg Serum DHT(pg/ml) Tissue T(ng/ml) Tissue DHT(pg/ml) Prostate wet weight (mg) Normal Castration Castrate + Testosterone Cheniton Nutmeg High Nutmeg Medium Nutmeg Low --2100400200100 1057.32±389.92218.26±258.35*1352.17±107.12673.24±203.31*632.21±149.34*715.82±229.37889.63±313.45 102.73±32.410.83±0.16*26.56±15.3233.65±22.0426.76±6.8437.93±11.8838.24±13.08 1043.90±279.872.45±1.41***961.44±158.67474.24±151.03**432.76±132.58**534.47±154.57*632.75±191.77 587.43±151.3918.23±8.91**933.72±301.75512.44±107.83*477.35±124.87*573.54±104.05*759.00±228.79

Note: Compared with castration + testosterone group, ^* P<0.05, ^** P<0.01, ^*** P<0.001

The results in Table 3 show that compared with the castration+testosterone group, each dose group of Nutmeg Extract C has a tendency to reduce serum DHT, tissue DHT and prostate wet weight, and the strength of the effect increases with the increase of concentration; The middle dose group has a significant effect on reducing serum DHT (P<0.05), tissue DHT (P<0.01) and prostate wet weight (P<0.05), and its effect strength is similar to that of Sernitone; the middle dose group has a significant effect on tissue DHT and prostate wet weight. Significant decrease (P<0.05).

Although the above-mentioned pharmacodynamics test is the drug efficacy test of the nutmeg extract C prepared in Example 2, the results of the administration test of different dosage groups have shown the dose-effect relationship of the drug extract of the present invention for treating benign prostatic hyperplasia, illustrating that the drug of the present invention It has an obvious effect on treating benign prostatic hyperplasia, and the effect is enhanced with the increase of administration dose, and can be used to prepare a pharmaceutical preparation for treating and/or preventing benign prostatic hyperplasia, with remarkable curative effect, stable quality and strong controllability.

Claims (10)

1, the purposes of Chinese medicine Semen Myristicae in preparation 5 inhibitor medicaments.

2, the purposes of Chinese medicine Semen Myristicae according to claim 1 in preparation 5 inhibitor medicaments, it is characterized in that: described 5 inhibitor medicaments is the medicine of treatment prostatic hyperplasia.

3, purposes according to claim 1 and 2, described Chinese medicine Semen Myristicae are the mature seed kernel of the tall and big aiphyllium plant of Myristicaceae Semen Myristicae Myristica fragrans Houtt..

4, purposes according to claim 1 and 2 is characterized in that, described Chinese medicine Semen Myristicae is Semen Myristicae protogenic medicinal powder or myristic water or extractive with organic solvent.

5, a kind of pharmaceutical composition that is used to prepare the 5 inhibitor, it is an active component by Semen Myristicae protogenic medicinal powder or its water/extractive with organic solvent, adds the preparation that acceptable accessories or complementary composition are prepared into.

6, the pharmaceutical composition that is used to prepare the 5 inhibitor according to claim 5 is characterized in that: described Semen Myristicae extractive with organic solvent is the extract of the mixed solvent of one or more solvent compositions in the organic solvents such as methanol, ethanol, ethyl acetate, n-butyl alcohol, ether, petroleum ether.

7, the pharmaceutical composition of preparation 5 inhibitor according to claim 5 is characterized in that: described preparation is oral formulations or injection.

8, a kind of method for preparing the described pharmaceutical composition of claim 5, wherein the preparation of Semen Myristicae extract may further comprise the steps:

A) Semen Myristicae water or 30～95% alcohol reflux, the solvent amount is 6～8 times, and extraction time is 1～3 hour, and merge extractive liquid,, concentrate drying get sample A;

B) A is water-soluble, with petroleum ether, ether, ethyl acetate or n-butanol extraction, decompression and solvent recovery gets water extract and corresponding extractive with organic solvent.

9, the purposes of each described pharmaceutical composition of claim 5～8 in preparation 5 inhibitor medicaments.

10, the purposes of each described pharmaceutical composition of claim 5～8 in preparation treatment prostatic hyperplasia medicine.