TW201600094A - An herbal composition against uterine fibroid and a use of its extract thereof - Google Patents

Abstract

An herbal composition against uterine fibroid is disclosed. The herbal composition comprises 25-75 wt% of Antrodia cinnamomea, 20.8-62.5 wt% of Astragalus onobrychis and 4.2-12.5 wt% of Leonurus japonicus. A use of an extract of the herbal composition for manufacturing drugs against uterine fibroid is also disclosed.

Description

For use in anti-uterine fibroids herbal compositions, and extracts thereof

The present invention relates to a Chinese herbal medicine composition, in particular to a herbal composition for anti-uterine fibroids, and to the use of the extract of the Chinese herbal medicine composition for preparing an anti-uterine fibroid drug.

Uterine fibroid is a benign tumor derived from the myometrium. It occurs in women between the ages of 30 and 50. Most uterine fibroids have no obvious symptoms, but with uterine fibroids. Growth, in addition to causing severe painful periods, abnormal gynecologic hemorrhage, may even lead to infertility, which can be achieved by uterine myomectomy Clear.

Conventional drugs are often used to improve the symptoms of uterine fibroids, such as the use of non-steroidal anti-inflammatory drugs (NSAID) to treat pain caused by uterine fibroids, or use oral contraceptive pill (or OCP) to reduce bleeding; in addition, gonadotropin-releasing hormone analog (GnRH analog) can reduce the amount of estrogen in the body to inhibit the growth of uterine fibroids. Long-term use of this GnRH analogue may cause symptoms such as osteoporosis or postmenopausal symptom, and uterine fibroids may continue to grow after discontinuation of administration, so it is only recommended to For specific patients.

Accordingly, it is still necessary to provide a Chinese herbal composition to combat the condition of uterine fibroids.

The main object of the present invention is to provide a herbal composition for anti-uterine fibroids which inhibits the growth of uterine fibroids.

Another object of the present invention is to provide a method for preparing an anti-uterine fibroids medicine by extracting the active ingredient of the Chinese herbal medicine composition, thereby improving the condition of the uterine fibroids, thereby improving the quality of life of the patient. By.

In order to achieve the foregoing object, the technical means and the effects achievable by the technical method include: a herbal composition for anti-uterine fibroids, comprising: 25 to 75 in weight percentage % of Antrodia camphorata, 20.8~62.5% of Astragalus, and 4.2~12.5% of Motherwort.

The herbal composition of the present invention, wherein the Chinese herbal medicine composition comprises 75% by weight of Antrodia camphorata.

The herbal composition of the present invention, wherein the Chinese herbal medicine composition comprises 20.8% by weight of xanthine.

The herbal composition of the present invention, wherein the Chinese herbal medicine composition comprises 4.2% by weight of motherwort.

The herbal composition of the present invention, wherein the Chinese herbal medicine composition comprises 75% by weight of Antrodia camphorata, 20.8% of Astragalus, and 4.2% of Motherwort.

The use of a Chinese herbal medicine extract for the preparation of an anti-uterine fibroids, wherein the Chinese herbal medicine extract is obtained by a method comprising the following steps: providing the method according to any one of claims 1 to 5 a Chinese herbal medicine composition; and extracting the Chinese herbal medicine composition with 95% alcohol at 50 to 80 ° C for 8 to 12 hours to obtain the Chinese herbal medicine extract; and concentrating the middle grass The drug extract; wherein the Chinese herbal medicine composition and the 95% alcohol have a weight-to-volume ratio of 1:2, the Astragalus membranaceus is a fruiting body, the scutellaria is a root, and the motherwort is an aerial part.

The use of the herbal extract of the present invention for preparing an anti-uterine fibroid drug, wherein the herbal extract is administered to a desired individual to prevent or ameliorate the uterine fibroid disorder of the desired individual, wherein the herbal extract The dose of the desired individual is administered to 300 mg of the herbal extract per kg of the desired individual body weight per day for 28 days.

The herbal composition for anti-uterine fibroids of the present invention can reduce the growth of uterine fibroids by reducing the content of eosin and lutein in the desired individual by using the composition ratio of Antrodia camphorata, Astragalus and Motherwort. .

The use of the herbal extract of the present invention for preparing anti-uterine fibroids is effective for inhibiting the growth of uterine fibroids by extracting active ingredients from Antrodia camphorata, Astragalus and Yimu, so that patients can be prevented from uterine fibroid growth. The dysmenorrhea, abnormal bleeding and other diseases, in order to improve the quality of life of patients.

Figure 1 is a H&E staining result of the uterine section of the group C0 rats.

Fig. 2 is a H&E staining result of the uterine horn section of the group C0 rats.

Fig. 3 is a H&E staining result of the uterine section of the group C1 rat.

Fig. 4 is a H&E staining result of the uterine horn section of the group C1 rat.

The above and other objects, features, and advantages of the present invention will become more apparent from the <RTIgt Antrodia cinnamomea , Astragalus onobrychis and Leonurus japonicus have the activity of inhibiting the growth of uterine fibroids by the composition ratio of Antrodia camphorata , Astragalus and Motherwort.

In detail, the Chinese herbal medicine composition comprises 25 to 75% by weight of Antrodia camphorata, 20.8 to 62.5% of Astragalus, and 4.2 to 12.5% of Motherwort, and the Chinese herbal composition preferably further comprises a weight. The percentage is 75% of Antrodia camphorata, 20.8% of Astragalus, and 4.2% of Motherwort. According to this, the Chinese herbal composition can inhibit the growth of uterine fibroids by a combination of various active ingredients, thereby eliminating the uterine muscle. Dysmenorrhea, abnormal bleeding and other diseases in patients with tumors.

Among them, the burdock is selected from the scorpion scorpion body, which is rich in triterpenoids; the scutellaria can choose the leguminous plant Astragalus membranaceus (Fisch.) Bge. Var. mongholicus (Bge.) Hsiao or the membrane pod The dry root of Astragalus membranaceus (Fisch.) Bge. is rich in glycosides, polysaccharides, flavonoids, amino acids and trace elements; the motherwort can choose the dry part of the plant, which contains Many alkaloids such as motherwort, motherwort, and stachysine. It also contains flavonoids such as benzoic acid, potassium chloride, lauric acid, linolenic acid, oleic acid, vitamin A and lanyl.

The herbal composition of the present invention can effectively inhibit the growth of uterine fibroids, and thus can be used as an active ingredient for assisting inhibition of uterine fibroids. Preferably, the Chinese herbal medicine composition can be extracted to obtain a Chinese herbal extract for preparation prevention. Or a medicament for treating uterine fibroids or a health food, the herbal extract being combined with a pharmaceutically acceptable carrier or excipient to form a pharmaceutical composition, wherein the herbal extract can be prepared into any convenient form. For example, a tablet, a capsule, a powder, a granule or a liquid, or the like, or a combination of the Chinese herbal extract and other foods or beverages, can be taken orally in a form suitable for consumption.

Wherein, the Chinese herbal medicine extract is preferably obtained by a method comprising the steps of: providing a herbal composition as described above; extracting the Chinese herbal composition with 95% alcohol at 50 to 80 ° C for 8 to 12 hours And extracting the extracted product to obtain the herbal extract.

In detail, the Chinese herbal medicine composition can be mixed with 95% alcohol in a weight ratio of 1:2, so that the various active ingredients enriched in the Chinese herbal composition can be dissolved in the 95% alcohol.

In the present embodiment, 500 grams of the Chinese herbal medicine composition is extracted, extracted with 1 liter of 95% alcohol, refluxed at 60 ° C for 10 hours, repeated extraction 3 times, and finally vacuum filtered, concentrated under reduced pressure and freeze-dried. To get the herbal extract.

Further, the herbal extract may be administered to a desired individual, for example, 300 mg of the herbal extract per kg of the desired individual per day, and continuously administered to the desired individual for 28 days. The active ingredient acts in the desired individual and inhibits the growth of uterine fibroids.

In order to confirm that the Chinese herbal medicine composition of the present invention does have anti-uterine fibroid activity, the following test was carried out:

( A ) Preparation of Chinese herbal extracts and detection of active ingredients

In this test, the composition ratio of the herbal composition of each group is as shown in Table 1, and the preparation method of the Chinese herbal medicine extract is as described above, and 500 g of each group of Chinese herbal medicine composition is taken, and 1 liter of 95% alcohol is used. After extraction, each group of herbal extracts can be obtained by filtration, concentration, and freeze-drying.

Weigh 0.2 g of each group of Chinese herbal medicine extract into a spiral test tube, add 5 mL of methanol, shake for 15 minutes with ultrasonic wave, centrifuge for 10 minutes at 3000 rpm, and take 5 mL. The supernatant was placed in a clean tube and heated to dryness in a 100 ° C water bath.

Continued analysis using a Purospher STAR (Merck) RP-18e (5 μm) 250 mm × 4 mm column, mixed with acetonitrile (Acetonitrile, ACN for short) and 0.085% phosphoric acid solution as a mobile phase at a flow rate of 1 mL / Min, the absorbance at a wavelength of 254 nm was detected, and the percentage of total triterpenes in each group was analyzed and recorded in Table 2.

In addition, galactose was used as a standard, and its concentration and absorbance were used to prepare a standard curve. Continue to configure each group of herbal extracts, take 1mL of each group of Chinese herbal extracts, add 1mL of 5% phenolic solution, then add 5mL of concentrated sulfuric acid, after standing for 10 minutes, shake it to make it evenly mixed, and measure by spectrophotometry The absorbance at 490 nm of each group was measured, and the concentration was calculated by a standard curve, and the results are recorded in Table 2.

According to the above test results, the herbal extracts of each group were rich in total triterpenoids and polysaccharides, and the content of group A3 was the highest.

( B ) Effect on survival of uterine sarcoma cells

This test system chosen human uterine sarcoma cells (MES-SA cells), commercially available from Food Industry Development Institute (BCRC 60333), the uterine sarcoma cell line cultures containing 1.5mM _L - glutamic acid _(L -Glutamine) and 10% Place 90% of the fetal bovine serum (FBS) in McCoy's 5a medium and place in a culture chamber at 37 ° C, 5% carbon dioxide and 95% humidity. Replace the fresh medium every two days.

When the uterine sarcoma cells are subcultured, the culture solution and the cells are centrifuged at 1000 rpm for 5 minutes, the supernatant is removed, a new culture solution is added, and the number of cells maintained in the 10 cm culture dish is about 1×. 10 ⁵ to 1 × 10 ⁶ cells/mL.

In the present experiment, the 10 cm culture dish of the above uterine sarcoma cells which have been 8 to 9 minutes old may be taken out, the discolored culture medium is removed, and the cells are washed once with 8 mL of PBS buffer solution, and trypsin/ethylenediamine is added. After acetonitrile (Trypsin/EDTA) reaction for 1~3 minutes, gently shake the culture dish to make the cells fall off the wall, add the warmed culture medium, gently disperse the cells, and then place the cell-containing culture solution into the centrifuge. After centrifugation at 1500 rpm for 10 minutes in the tube, after removing the supernatant, the serum-containing culture solution was added to uniformly disperse the cells, and 20 μL of the cells were taken out, and 20 μL of 0.04% trypan blue (Trypan Blue) was added for staining. Into the cell counter, and count the number of viable cells under the microscope, the cell survival rate must be not less than 85% before the subsequent test can be carried out.

The concentration of the above uterine sarcoma cells was adjusted to 1×10 ⁵ cells/mL with a serum-containing culture solution, and 100 μL of each uterine sarcoma cell was seeded in a 96-well plate, so that each well contained 1×10 ⁴ cells at a temperature of Incubate overnight at 37 ° C in an incubator with a carbon dioxide concentration of 5%.

After 24 hours of incubation, add 100 μL of the herbal extract (concentration: 2 mg/mL, dissolved in DMSO) as shown in Table 3, mix well, and incubate at a temperature of 37 ° C and a carbon dioxide concentration of 5%. Continue to train for 24 hours overnight.

After 24 hours, remove the culture solution, rinse with phosphate buffer, then add 100 μL contains fresh culture medium of CCK-8, and after being reacted for 2 hours in an incubator at a temperature of 37 ° C and a carbon dioxide concentration of 5%, after shaking for 5 minutes, the absorbance of each well at a wavelength of 450 nm is detected. The survival rate was calculated based on the absorbance results of the control group without any drug treatment and is recorded in Table 3.

Please refer to Table 3, according to the above test results, the Chinese herbal extract does have the ability to inhibit the growth of uterine sarcoma cells, and the Chinese herbal extracts of Group A3 have the best efficacy, so that the A3 group of Chinese herbal medicines can be reused. The extract was subjected to subsequent tests.

( C ) Effect on uterine sarcoma rats

In this experiment, Wistar female rats purchased from the National Experimental Animal Center of the National Experimental Research Institute of the Foundation were selected. The rats were all over 8 weeks old and weighed between 180 and 190 grams. The rats were kept in the animal center of Daren University of Science and Technology, and the animal room was kept at room temperature of 20-22 ° C. The light time and dark time were 12 hours each. The rats were free to eat and drink.

RO water (Group C0) or Group A3 herbal extract (Group C1) was administered to the rats via a gastric tube (administered at a dose of 300 mg of herbal extract per kilogram per day), while borrowing Estrogen was administered by intraperitoneal injection (dose was 200 μg/time), and the body weight, food intake and water intake of each group were recorded daily. After 4 weeks, blood was collected from the celiac artery to measure the emodin in the blood. The content of progesterone was taken out, and the uterus was taken out for tissue sectioning and H&E staining (hematoxylin-eosin). The results are shown in Table 4 and Figure 1-4, respectively.

Please refer to Table 4 for the weight and feeding of rats in each group of this test after 28 days. There was no significant difference in the amount of water and the amount of drinking water, and the levels of eosin and lutein in the blood of the C1 group of the Chinese herbal extracts were significantly decreased (the inhibition rate of the emoji was 47.5±4.8%, but The inhibition rate of lutein was 33.1±5.1%, indicating that the Chinese herbal extract did have the ability to reduce the levels of emodin and lutein in the body.

Continued with reference to Figures 1 and 2, respectively, the results of tissue section staining of the uterus and uterine horn of the group C0, the uterine smooth muscle cell line showed hypertrophy, with eosinophilic cytoplasm, cell boundaries are not clear, The arrangement was staggered and disordered, and cell mitosis was not observed, indicating that the uterine fibroids were successfully induced in the group C0. Further, please refer to the figures of Figures 3 and 4, and the results of tissue staining of the rats in the C1 group showed that Oral administration of the Chinese herbal extract can successfully inhibit uterine smooth muscle hyperplasia and hypertrophy, indicating that the Chinese herbal extract does have an activity of improving uterine smooth muscle hyperplasia and hypertrophy.

In summary, the herbal composition for anti-uterine fibroids of the present invention can reduce the emollient and lutein content in each body by reducing the composition of the body composition of the burdock, scutellaria and motherwort. The effect of tumor growth.

Furthermore, the use of the herbal extract of the present invention for preparing anti-uterine fibroids is effective for inhibiting the growth of uterine fibroids by extracting active ingredients from Antrodia camphorata, Astragalus and Yimu, so that patients can be free of uterine muscles. Tumor growth caused by dysmenorrhea, abnormal bleeding and other conditions, and thus improve the quality of life of patients.

While the invention has been described in connection with the preferred embodiments described above, it is not intended to limit the scope of the invention. The technical scope of the invention is protected, and therefore the scope of the invention is defined by the scope of the appended claims.

Claims (7)

The invention relates to a herbal composition for anti-uterine fibroids, which comprises: 25 to 75% by weight of Antrodia camphorata, 20.8 to 62.5% of Astragalus, and 4.2 to 12.5% of Motherwort. The Chinese herbal medicine composition according to claim 1, wherein the Chinese herbal medicine composition comprises 75% by weight of Antrodia camphorata. The Chinese herbal medicine composition according to claim 1, wherein the Chinese herbal medicine composition comprises 20.8% by weight of xanthine. The herbal composition according to claim 1, wherein the herbal composition comprises 4.2% by weight of motherwort. The Chinese herbal medicine composition according to claim 1, wherein the Chinese herbal medicine composition comprises 75% by weight of Antrodia camphorata, 20.8% of Astragalus, and 4.2% of Motherwort. The use of a Chinese herbal medicine extract for the preparation of an anti-uterine fibroids, wherein the Chinese herbal medicine extract is obtained by a method comprising the following steps: providing the method according to any one of claims 1 to 5 a Chinese herbal medicine composition; the Chinese herbal medicine composition is extracted with 95% alcohol at 50 to 80 ° C for 8 to 12 hours; and the extracted product is concentrated to obtain the Chinese herbal medicine extract; wherein the Chinese herbal medicine composition and the 95% alcohol The weight-to-volume ratio is 1:2, and the Astragalus membranaceus is a fruiting body, the scutellaria is a root, and the motherwort is an aerial part. The use of the herbal extract according to claim 6 of the patent application for preparing an anti-uterine fibroid drug, wherein the herbal extract is administered to a desired individual to prevent or ameliorate the uterine fibroid disorder of the desired individual, Wherein, the Chinese herbal medicine extract is administered to the desired individual at a dose of 300 mg of the herbal extract per kg of the desired individual body weight per day for 28 days.