TWI462742B - A use of an herbal extract for manufacturing drugs against sarcoma - Google Patents

Description

Use of Chinese herbal extract to prepare a medicament for inhibiting the growth of sarcoma cells

The present invention relates to a compound composition of Antrodia camphorata, in particular to a combination of Ganoderma lucidum and licorice, which can inhibit the growth of tumor cells, and the present invention relates to a pharmaceutical composition comprising the complex formula of Antrodia camphorata.

The treatment of tumors is different in Chinese and Western medicine. Western medicine mainly uses surgery, radiation therapy, chemotherapy and immunotherapy; Chinese medicine uses some traditional Chinese herbal medicines as treatment.

Antrodia (Taiwanofungus camphoratus), also known as mushrooms or Antrodia camphorata, based belonging Basidiomycota (Basidiomycota) in Fungi, Basidiomycotina (Basidiomycotina), with Basidiomycetes (Homobasidiomycetes), no off bacteria mesh (Aphyllophorales ), Polyporaceae , Antrodia , Antrodia , which grows only on the inner wall of hollow core material of Taiwan's endemic burdock tree ( Cinnamomum kanehirai ). Its fruit bodies are annual to perennial, bright red and orange. Or light cinnamon, with a scent of eucalyptus.

From the mycelium or fruit body extract of Antrodia camphorata, a variety of physiologically active substances such as polysaccharides, adenosine or several triterpenoids are found, among which triterpenoids, such as Aminocin A, Aminocin B (Antcin B), Amino C (Antcin C), Amine E (Antcin E), Amine F (Antcin F), Methylantcinate G, Methylamine oxime H ( Methylantcinate H), Antrocin, 4,7-bismethoxy-5-methyl-1,3 dioxygen Benzene (4,7-dimethoxy-5-methyl-1,3-benzodioxole) and 2,2',5,5'-tetramethoxy-3,4,3',4'-bismethyl-internal double Oxygen-6,6'-biphenyl (2,2',5,5'-tetramethoxy-3,4,3',4'-bimethyl-enedioxy-6,6'-dimethylbiphenyl), etc. The ability of tumor cells to grow.

In the classification of traditional Chinese medicine, Antrodia camphorata is a cold-cold medicine. Although it has excellent anti-inflammatory effect under short-term use, it has better health effects for chronic hepatitis caused by improper diet or long-term fatigue. However, long-term use Antrodia camphorata may cause excessive concentration of free radicals in the body, resulting in a decline in body constitution and decreased immunity. For patients with cancer treated with chemotherapy or radiation therapy, the initial use of Antrodia camphorata can reduce side effects, but long-term use of high doses of Antrodia camphorata, On the contrary, it may cause a decrease in immunity such as an increased risk of blood infection.

The main object of the present invention is to provide a compound formula of Antrodia camphorata which has better tumor suppressing effect than Niobium sinensis.

A further object of the present invention is to provide a compound formula of Antrodia camphorata which can alleviate side effects such as qi deficiency caused by long-term use of Antrodia camphorata.

Still another object of the present invention is to provide a pharmaceutical composition comprising the Astragalus lucidum compound formulation as a drug component for inhibiting growth of tumor cells.

In order to achieve the foregoing object, the technical means and the efficiencies achievable by the technical method include: a compound composition of Antrodia camphorata, comprising: 27.3 to 50.0% by weight of Antrodia camphorata, by weight percentage Ganoderma lucidum with a ratio of 27.3 to 40.0% and licorice at a weight ratio of 16.7 to 45.4%.

The composition of the Antrodia camphorata of the present invention comprises 30.8% by weight of Antrodia camphorata.

The composition of the Antrodia camphorata of the present invention comprises 30.8% by weight of Ganoderma lucidum.

The Astragalus lucidum composite formulation of the present invention comprises 38.5% by weight of licorice.

The composition of the Antrodia camphorata of the present invention comprises 30.8% by weight of Antrodia camphorata, 30.8% by weight of Ganoderma lucidum and 38.5% by weight of licorice.

In the complex formula of the Antrodia camphorata of the present invention, the Antrodia camphorata is selected from the group consisting of Antrodia camphorata fruiting bodies.

A pharmaceutical composition comprising: a combination of anthraquinone as described above; and a pharmaceutically acceptable carrier or excipient.

The complex formula of the Antrodia camphorata of the present invention can effectively inhibit the growth of tumor cells, and thus can serve as an auxiliary component for inhibiting tumor cells, and is an effect of the present invention.

The composite formula of the Antrodia camphorata of the invention can effectively inhibit the growth of tumor cells compared with the single prescription of Antrodia camphorata, Ganoderma lucidum or licorice, and can achieve the effect of slowing down the side effects caused by long-term use of Antrodia camphorata through the composition ratio of various Chinese herbal medicines. .

The pharmaceutical composition comprising the complex composition of the Antrodia camphorata is effective for inhibiting the growth of tumor cells by the complex formula of Antrodia camphorata, and has the effect of inhibiting the proliferation of tumor cells and preventing proliferation or spread.

Figure 1A is a graph showing the cell viability of mouse sarcoma cells treated with the complex formula of Antrodia camphorata.

Figure 1B is a histogram of cell viability of mouse sarcoma cells treated with 125 μl/ml of the Astragalus membranaceus complex formulation of the present invention.

Figure 2A is a graph showing the cell viability of human lung tumor cells treated with the complex formula of Antrodia camphorata.

Figure 2B is a bar graph of cell viability of human lung tumor cells treated with the 125 μl/ml of the Astragalus membranaceus complex formulation of the present invention.

Fig. 3A is a graph showing the cell survival rate of human liver tumor cells treated with the complex formula of Antrodia camphorata of the present invention.

Figure 3B is a bar graph of cell viability of human liver tumor cells treated with the 125 μl/ml of the Antrodia camphorata complex formulation of the present invention.

Fig. 4A is a graph showing the cell survival rate of human colorectal tumor cells treated with the complex formula of the Antrodia camphorata of the present invention.

Figure 4B is a bar graph of cell viability of human colorectal tumor cells treated with 125 μl/ml of the Astragalus membranaceus complex formulation of the present invention.

Fig. 5A is a graph showing the cell survival rate of human breast tumor cells treated with the complex formula of Antrodia camphorata of the present invention.

Figure 5B is a bar graph of cell viability of human breast tumor cells treated with the 125 μl/ml of the Astragalus membranaceus complex formulation of the present invention.

Fig. 6A is a diagram showing the size of a mouse sacrificed after 30 days of feeding a tumor mouse with the Astragalus membranaceus complex preparation of the present invention.

Fig. 6B is a graph showing the tumor growth in mice fed tumor mice by the complex formula of the anatum.

Fig. 7 is a survival line chart of a mouse fed a tumor mouse with the complex formula of the Antrodia camphorata of the present invention.

The above and other objects, features, and advantages of the present invention will become more apparent from the aspects of the invention. Including Ganoderma lucidum and licorice, the Antrodia camphorata, the Ganoderma lucidum and the licorice are mixed in a specific ratio, in particular, 27.3 to 50.0% by weight of Antrodia camphorata, 27.3 to 40.0% by weight of Ganoderma lucidum and by weight The ratio of 16.7 to 45.4% of licorice is effective in inhibiting the growth of tumor cells.

In the preferred embodiment, the body of the burdock is used, and the triterpenoids of the body of the burdock are high in content, and the anti-inflammatory effect is better; in addition, the preferred embodiment adopts the body of Ganoderma lucidum and the licorice root and rhizome. The above-mentioned parts are relatively easy to obtain, and thus the cost can be reduced, but not limited thereto.

The combination formula of the Antrodia camphorata of the present invention can effectively inhibit the growth of tumor cells, and thus can be used as an auxiliary component for inhibiting tumor cells, and preferably the extract of the Astragalus membranaceus compound formula can be used for preparing a medicament for preventing or treating tumors or The health food, the extract of the anathopin compound formula and the pharmaceutically acceptable carrier or excipient are combined to form a pharmaceutical composition, wherein the extract of the anathopin compound formula can be prepared into any convenient type, such as A tablet, a capsule, a powder, a granule or a liquid, or the like, or a combination of the Chinese herbal compound extract and other foods or beverages, for oral administration to a living body.

In order to confirm that the Astragalus lucidum composite formulation of the preferred embodiment of the present invention has a better tumor suppressing effect, the following test is carried out:

(A) Preparation of crude extract of Antrodia camphorata compound formula

In this test, the Chinese herbal medicines shown in Table 1 are mixed. Preferably, the Chinese herbal medicines can be broken into smaller particles, for example, less than 5 mm square particles, and then mixed. Increase its mixing efficiency and subsequent extraction efficiency.

Table 1, the composition of each group of alcohol extracts and their weight percentage (%)

Take 5 kg of each group of mixed Chinese herbal medicines, extract with 1 liter of 95% alcohol, reflux at 50 ° C for 8 hours, and repeat the extraction three times. Finally, by vacuum filtration, concentration under reduced pressure and freeze-drying, A crude alcohol extract was taken and a subsequent test was conducted with the crude alcohol extract.

(B) Effect on survival rate of tumor cell lines

This experiment used mouse sarcoma (S-180 cells), human lung tumors (A549 cells), human liver tumors (HepG2 cells), human colorectal tumors (HCT-116 cells) and human breasts purchased from the Food Industry Development Research Institute. Tumor (MDA-MB-231 cells) cell lines, all of which were cultured in 10% fetal bovine serum (FBS, Biological Industries, Kibbutz beit haemek), 2 mmol/L _L -glutamic acid ( _L- Glutamine, Culture medium purchased from HyClone, USA), 1 x non-essential amino acid (purchased from HyClone, USA), 100 μg/ml streptomycin, and 100 U/ml penicillin (Penicillin, available from HyClone, USA) (Dulbecco's Modified Eagle Medium; DMEM), the tumor cells were placed in an incubator at 37 ° C, 5% carbon dioxide, and 95% humidity, and the fresh medium was replaced every two days.

When the above-mentioned tumor cell strain is subcultured, the culture solution and the cells can be centrifuged at 1000 rpm for 5 minutes, the supernatant is removed, then the fresh 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.

When the test is carried out, the 10 cm culture dish of the above tumor cell line which has been 8 to 9 minutes old may be taken out, the discolored culture medium is removed, the cells are washed by adding 8 ml of PBS buffer solution, and trypsin/ethylenediamine acetic acid is added. Trypsin/EDTA), after 1 to 3 minutes of reaction, gently shake the culture dish to detach the cells from the wall, add the warmed culture medium, gently disperse the cells, and then place the cell-containing culture solution into the centrifuge tube. After centrifugation at 1500 rpm for 10 minutes, after removing the supernatant, the serum-containing culture solution was added to uniformly disperse the cells, and 20 μl of the tumor cells were taken out, and 20 μl of 0.04% trypan blue (Trypan Blue) was added for staining. , put into the cell counter, and count the number of viable cells under the microscope, the cell survival rate must be not less than 95% before the subsequent test can be carried out.

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

After 24 hours of incubation, the crude alcohol extract as shown in Table 1 was added, mixed well, and cultured overnight at 37 ° C in a 5% carbon dioxide incubator for 24 hours.

After 24 hours, the culture solution was removed, rinsed with phosphate buffer, and then treated with 0.5 mg/ml MTT for 4 hours, and the absorbance of each well at a wavelength of 570 nm was detected by a spectrophotometer.

Please refer to Figure 1A for the cell viability curve of sarcoma cells treated with different concentrations of alcohols in groups A1 to A7. The survival rate (%) is calculated as follows: survival rate = (test group Absorbed value measured / absorbance measured in the control group) × 100%

It can be known from the experimental results that the composite formula of the Antrodia camphorata of the preferred embodiment (the curve of the A1 to A4 group) is compared with the single side of the Antrodia camphorata, Ganoderma lucidum and licorice (the curve of the A5 to A7 group). It does have a better mouse sarcoma cell survival inhibitory effect.

Please refer to Figure 1B for the cell viability map of mouse sarcoma cells treated with crude alcohol extracts of groups A1 to A7 at a concentration of 125 μl/ml. Group A3 has a better inhibition of mouse sarcoma cell survival. effect.

In addition, both human lung tumor cells (as shown in Figures 2A and 2B), human liver tumor cells (as shown in Figures 3A and 3B), human colorectal tumor cells (as shown in Figures 4A and 4B) and Human breast tumor cells (as shown in Figures 5A and 5B) have a composite formulation of the Antrodia camphorata (Groups A1 to A4) of the preferred embodiment compared to O. chinensis, Ganoderma lucidum and licorice unilateral (Groups A5 to A7). A preferred survival inhibition effect.

(C) Evaluation of tumor suppressive efficacy in tumor mice

This trial used Balb/C male mice purchased from the Animal Center of the National Cheng Kung University School of Medicine. The mice were all over 8 weeks old and weighed between 20-25 g. The mice were housed in the SPF Animal Center of the University of Stanley, and maintained in an animal room at room temperature of 25 ± 1 °C for 12 hours each. Moreover, mice were free to eat and drink; feed (purchased from LabDiet, Inc., USA) and drinking water were supplied by the Animal Center of the University of Science and Technology.

The mouse sarcoma cell strain (such as the aforementioned S-180 cell strain, purchased from the Food Industry Development Research Institute) was diluted with physiological saline to a concentration of 5×10 ⁶ cells/ml, and the mouse sarcoma cell strain was small. The back of the mouse was injected subcutaneously.

Next, the crude alcohol extract of the A3 group was diluted in the mouse feed at a ratio of 0.25% (Group C2), 0.5% (Group C3), and 1% (Group C4), respectively. The C1 group of the crude alcohol extract of the preferred embodiment was used as the control group, and the mice were allowed to eat for 30 days freely, and the tumor size was measured every 3 days (tumor size calculation formula = length × width × height × 0.45), and each observation was observed. The survival rate of the mice in the group.

Referring to Figures 6A and 6B, the addition of the crude alcohol extract of the preferred embodiment (Groups C2 to C4) to the mouse feed can effectively reduce the volume of the tumor and its growth rate. The degree (as shown in Fig. 6B), and the inhibitory effect of tumor growth is positively correlated with the concentration of the crude alcohol extract of the preferred embodiment.

Furthermore, referring to Figure 7, feeding the crude extract of the preferred embodiment can also significantly reduce the lethality of tumor cells to mice.

In summary, the complex formula of the Antrodia camphorata of the present invention can effectively inhibit the growth of tumor cells, and thus can serve as an auxiliary component for inhibiting tumor cells, and is an effect of the present invention.

Furthermore, the composite formula of the Antrodia camphorata of the present invention has a better inhibitory effect on tumor cells than the single-side of Antrodia camphorata, Ganoderma lucidum or licorice, and can reduce the long-term use of the unilateral use of Antrodia camphorata through the composition ratio of various Chinese herbal medicines. The efficacy of side effects.

In addition, the pharmaceutical composition comprising the complex composition of the Antrodia camphorata effectively inhibits the growth of tumor cells by the complex formula of Antrodia camphorata, and has the effect of inhibiting the proliferation of tumor cells and preventing proliferation or spread.

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 (1)

A Chinese herbal medicine extract for preparing a medicament for inhibiting the growth of sarcoma cells, wherein the herbal extract is diluted in a diet of 0.25 to 1% in a diet of a desired individual, so that the desired individual is free to ingest for 30 days to inhibit The growth of sarcoma cells in the desired individual; wherein the herbal extract is obtained by the following steps: providing a mixture of Chinese herbal medicines comprising: 30.8% by weight of Antrodia camphorata, 30.8% of Ganoderma lucidum and 38.5 % of licorice; extracting the Chinese herbal mixture with 95% alcohol at 50 ° C for 8 hours, repeating the extraction three times, the weight ratio of the herbal mixture to the alcohol is 5:1; and vacuum filtration, concentration under reduced pressure and freeze drying, To obtain the herbal extract.