TWI430792B - Pharmaceutical composition for treating hepatitis and/or cancer - Google Patents

Description

Pharmaceutical composition for treating hepatitis and/or cancer

The present invention relates to pharmaceutical compositions and methods for treating hepatitis and/or cancer.

Hepatocellular carcinoma is a common cancer in Taiwan and Asia and Africa. The prevalence is 5 to 10 times that of Europe and America, and its mortality rate is almost 100%. Take Taiwan as an example, it accounts for the first cause of death in male malignant tumors and the third leading cause of death in female malignant tumors. After general discovery, even after surgery, chemotherapy or radiation therapy, the average life expectancy is less than one year.

According to epidemiological statistics, high-infection areas of hepatitis virus around the world have high cases of liver cancer, such as Taiwan, China, Southeast Asia and South Africa. Especially for hepatitis B, if Taiwan is taken as an example, more than 90% of liver cancer cases are the carriers of hepatitis B, and the chance of getting liver cancer instead of the original one is only the percentage of the original. One (Beasley, RP, 1988, Cancer, 61, 1942-1956; Beasley, RP, et al., 1981, Lancet, 2, 1129-1133).

In the past, liver cancer caused by hepatitis B virus infection was only transformed into cancer cells by many regulatory mechanisms of hepatocytes. Nowadays, scholars believe that the genes contained in B virus itself and the proteins they express are important "carcinogenic factors" ( Arthran-Ruiz, P., et al., 2002, J Gen Virol, 83, 2059-2073; Caselmann, WH et al., 1990, PNAS, 87, 2970-2974; Hildt, E., et al., 1996 , Virology, 225, 235-239; Hildt, E., et al., 1996, Hepatology, 24, 502-507; Hildt, E., et al., 1995, Oncogene, 11, 2055-2066; Hildt, E., et al , 1993, Oncogene, 8, 3359-3367; Huang, TJ, et al., 2005, J Biol Chem, 280, 27742-27754; Kekule, AS, et al., 1990, Nature, 343, 457-461). Drugs used clinically to treat viral hepatitis are currently treated with interferon alpha 2a (Ola, SO, et al., 2000, West Afr J Med, 19259-64) and nucleic acid analogs (lamivudine). Known as hepatic energy) (Grossi, P., et al., 2001, Transplant Proc, 33, 1576-1578; Matsuo, K., et al., 2001, Leuk Lymphoma, 41, 191-195), but the results It is still limited and even produces resistance.

In recent years, the development of liver disease drugs has gradually developed towards Chinese herbal medicines, such as Phyllanthus urinaria L. (Calixto, JB, et al., 1998, Med Res Rev, 18, 225-258; Huang, TJ, et al ., 2003, Phytother Res, 17 449-453; Mahdavi, J., et al., 2002, Science, 297, 573-578; Wang, BE, 2000, J Gastroenterol Hepatol, 15, E67-70) and Piper Kadsura ) (Huang, RL, et al., 2001, J Chin Med, 12, 179-190), Terminalia chebula Retz. , Sanguisorba officinalis L. , Rubus coreanus Miq. The natural component extracted from Rheum palmatum L. (Kim, TG, et al., 2001, Phytother Res, 15, 718-720) was also found to have an anti-hepatitis B virus effect. But apart from this, there is very little literature on the study of hepatitis virus in Chinese herbal medicines.

Solanaceae plants (Solanaceae) is the world's third most important cash crop, is currently the most important vegetable crops, such as fruits and vegetables (tomatoes, eggplant, peppers, sour tomato sauce ... etc), eating leafy vegetables (Solanum aethiopicum, S. Macrocarpon ) and medicinal plants (such as Datura , Capsicum ). Solanaceae contains a variety of chemical components, such as alkaloids, polyphenols and sterols (Arthan, D., et al., 2006, Phytochemistry, 67, 27-33; Caceres, A., et al . 1998, J Ethnopharmacol, 62, 195-202; Jung, K., et al., 2005, Arch Pharm Res, 28, 1381-1385; Lee, DG, et al., 2005, Arch Pharm Res, 27, 1031 1036; Parr, AJ, et al., 2005, J Agric Food Chem, 53, 5461-5466), which has been reported to have antibacterial, anti-oxidant, anti-ulcer or anti-cancer activities (Caceres, A., Et al., 1998, J Ethnopharmacol, 62, 195-202; Gan, KH, et al., 1993, J Nat Prod, 56, 15-21; Iwalewa, EO, et al., 2005, J Med Food, 8, 539-544 ;Lee, DG, et al., 2005, Arch Pharm Res, 27, 1031-1036), but no studies have been conducted on the use of Solanaceae plants for the inhibition of hepatitis viruses.

Hepatocellular carcinoma is a common cancer in Asia and Africa. After surgery, even after surgery, chemotherapy or radiation therapy, the average survival rate is not high. According to epidemiological statistics, there are high cases of liver cancer in the high-infection areas of hepatitis virus around the world. Taking Taiwan as an example, more than 90% of liver cancer cases are carriers of hepatitis B. In addition, there has been evidence in the academic community that hepatitis virus infection can cause liver cancer, and the genes themselves and the proteins they present are also important carcinogenic factors. Solanaceae plants contain a variety of chemical components such as alkaloids, polyphenols and sterols, which have antibacterial, anti-oxidant, anti-ulcer or anti-cancer activities, but no predecessor technology has applied Solanaceae plants to inhibit hepatitis. Above the use of the virus. It is therefore an object of the present invention to develop a method of treating hepatitis and cancer using a pharmaceutical composition extracted from Solanaceae.

The term "or" in this document is also the same as "and/or".

In the following, the terms "a" or "an" are used to describe the elements and components of the invention, and the basic concepts of the invention are given for convenience only. This description is to be construed as inclusive of the singular

The term "patient" below refers to a person who is undergoing or requiring health or medical care and/or treatment. Among them, the person may be waiting for this care or may be receiving this care, or may have accepted this care.

The following term "Solanaceae (Solanaceae)" means all plants in the nightshade family, including fresh fruit and vegetables, include: root potato, tomato, eggplant, peppers, tomato sauce or acid such as; eating leafy vegetables (Solanum aethiopicum, S . macrocarpon); and / or medicinal plants, including: Datura (Datura) plant, green pepper genus (Capsicum) plants, which in turn contain natural mountain are not limited to tobacco, eggplant, India, Wan peach, or large flower mandala and so on.

The term "hepatitis virus carcinogenic factor" as used hereinafter means the gene itself carried by the hepatic virus itself and the protein it expresses, which may alter the normal physiological functions of human cells and/or cause cancer.

The term "Toxicity" as used hereinafter means that cancer cells are treated with a pharmaceutical composition, cell growth is inhibited, stagnated, or the number of cancer cells caused by apoptosis or death is reduced.

The term "pharmaceutically acceptable carrier" as used hereinafter means that the pharmaceutical composition and the medicinal composition are acceptable according to the conventional pharmaceutical combination technology for the needs of the pharmaceutical manufacturing process, for facilitating drug dosage formulation, or for different dosage forms. Bulk carrier mixing, suitable pharmaceutical acceptable carriers are well known in the art, and the carrier can be in a wide variety of forms. Certain pharmaceutical acceptable carriers can be found in The Hand Book of Pharmaceutical Excipients, published by the American Pharmaceutical Association and the Pharmaceutical Society of Great Britain. For example, a tablet, capsule, gel, solution or suspension may also contain the following ingredients: a pharmaceutically acceptable excipient or carrier which is non-toxic, inert solid or semi-solid, diluent, encapsulating material ( Encapsulating material), gel base or any form of formulation adjuvant, such as: microcrystalline cellulose, glucose, skim milk powder, starch, vermiculite, anhydrous calcium hydrogen phosphate, magnesium phosphate, stearic acid, hard Magnesium fatty acid, or artificial flavors and other substances.

The present invention provides a pharmaceutical composition for treating hepatitis, the pharmaceutical composition comprising a plurality of linoleic acid compounds, or a pharmaceutically acceptable salt thereof, the linoleic acid compound having the structural formula of Formula I, wherein R H, the structure is as follows:

In a specific embodiment, the pharmaceutical composition of the present invention further comprises a pharmaceutically acceptable carrier.

The present invention also provides a method of preparing a pharmaceutical composition of the present invention. In one embodiment, the pharmaceutical composition is extracted from a Solanaceae plant, and in another embodiment, the pharmaceutical composition is The preparation method is obtained by extracting and concentrating the Solanaceae plant by repeated organic solvent extraction and extracting by column chromatography. In yet another embodiment, the Solanaceae plant includes, but is not limited to, mountain tobacco. In another embodiment, the method of preparing the pharmaceutical composition is obtained by extracting and concentrating the mountain tobacco by repeating several times of organic solvent extraction and using column chromatography.

The present invention further provides a method for treating hepatitis, which is caused by the hepatitis-hepatitis virus infection in a specific embodiment of the present invention, and in a specific embodiment of the present invention, the hepatitis virus is a hepatitis B virus.

In a particular embodiment of the invention, the method of treating hepatitis comprises administering to a patient infected with a hepatitis virus a therapeutically effective amount of a pharmaceutical composition of the invention, the therapeutically effective dose being the necessary dose to elicit an ideal biological effect.

The invention further provides a method of inhibiting hepatitis virus. In a specific embodiment of the present invention, the pharmaceutical composition of the present invention is effective for inhibiting gene replication of hepatitis virus in infected cells. In a preferred embodiment, the hepatitis virus is hepatitis B virus and is in another In a specific embodiment, the pharmaceutical composition of the invention reduces the expression of hepatitis B virus protein in infected cells, and in a preferred embodiment, the protein is hepatitis B virus surface antigen (HBsAg). In yet another embodiment, the pharmaceutical composition of the present invention inhibits secretion of hepatitis B virus protein by infected cells. In a preferred embodiment, the protein is hepatitis B virus surface antigen (HBsAg) and/or e antigen (HBeAg).

The present invention also provides a pharmaceutical composition for treating cancer, the pharmaceutical composition comprising a plurality of linoleic acid compounds, or a pharmaceutically acceptable salt thereof, the linoleic acid compound having the structural formula I, wherein the R system H, the structural formula is as follows:

In a specific embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

The present invention also provides a method of preparing a pharmaceutical composition of the present invention. In one embodiment, the pharmaceutical composition is extracted from a Solanaceae plant, and in another embodiment, the pharmaceutical composition is The preparation method is obtained by extracting and concentrating the Solanaceae plant by repeated organic solvent extraction and extracting by column chromatography. In yet another embodiment, the Solanaceae plant includes, but is not limited to, mountain tobacco. In another embodiment, the method of preparing the pharmaceutical composition is obtained by extracting and concentrating the mountain tobacco by repeating several times of organic solvent extraction and using column chromatography.

The invention further provides a method for treating cancer. In a specific embodiment, the cancer is selected from the group consisting of brain, lung, squamous cell, bladder, stomach, pancreas, liver, breast, head, neck, kidney, kidney, ovary, In the preferred embodiment of the invention, the prostate, the esophagus, the blood, the bone, the sacrum, the gynecological or the thyroid cancer, etc., the cancer is not limited to liver cancer, and in another specific embodiment, the liver cancer is from hepatitis. In another embodiment, the hepatitis is caused by a hepatitis virus infection, and in a specific embodiment, wherein the hepatitis virus is a hepatitis B virus.

In a particular embodiment of the invention, the method of treating hepatitis comprises administering to a patient infected with a hepatitis virus a therapeutically effective amount of a pharmaceutical composition of the invention, the therapeutically effective dose being the necessary dose to elicit an ideal biological effect.

In a specific embodiment of the present invention, the pharmaceutical composition of the present invention inhibits chronic hepatitis caused by hepatitis B virus infection, and further reduces the occurrence of liver cancer induced by chronic hepatitis. In another embodiment of the present invention, the pharmaceutical composition of the present invention can also inhibit the expression and/or secretion of the hepatitis B virus carcinogenic factor, thereby further reducing the chance of liver cancer.

Further, in a specific embodiment of the present invention, the pharmaceutical composition of the present invention has a toxic effect on liver cancer cells. In a preferred embodiment, the pharmaceutical composition is administered in a high dose.

The invention may be embodied in different forms and is not limited to the examples mentioned below. The following examples are merely representative of the different aspects and features of the present invention.

Example 1: Extraction method of pharmaceutical composition of the present invention

As shown in Figure 1, the dried leaves of Solanum erianthum have a dry weight of 0.8-2.4 kg (kg), and are extracted with methanol (Methanol) three times at room temperature. The obtained extract is concentrated by a vacuum condenser. Thereafter, after removing the n-hexane layer using 2 liters (L) of n-hexane ( n- Hexane): methanol:water (H ₂ O) = 4:3:1, liquid partitioning, the remaining liquid concentrated under reduced pressure, and then with water H ₂ O: 1 for dispensing a liquid phase, the ethyl acetate layer to obtain 62 g (G), via the n-hexane:: ethyl acetate (ethyl acetate) = 1 ethyl acetate = 20: 1 Gradient bank is used to collect the first to 18th extracts (SEL1~18), and SEL9 is separated by Sephadex LH-20 to obtain three liquid separation layers (SE9). -1~3), wherein the third liquid layer (SE9-3) is further purified by a gradient of n-hexane:acetone (Acetone)=50:1 to obtain 15 liquid separation layers, and then take the 9th (SE9-3- 9) The liquid separation layer was subjected to dichloromethane: Dichloromethane: methanol = 2:1 to obtain 4 liquid separations through a Sephadex LH-20 column. Thereafter, the fourth liquid separation layer (SE9-3-9-4) was extracted through an open column with a gradient of n-hexane:acetone=10:1 to obtain four liquid separations. The fourth liquid separation layer (SE9-3-9-4-4) separated above was concentrated under reduced pressure to obtain a pharmaceutical composition of the present invention, and subjected to nuclear magnetic resonance spectroscopy (NMR): ¹ H-NMR analysis of the spectrum (as shown in Figure 2) and ¹³ C-NMR analysis spectra (shown in Figure 3), the measured values are shown in Table 1:

From the above-mentioned nuclear magnetic resonance spectrum data analysis, it is shown that the pharmaceutical composition of the present invention has the structural formula of the linoleic acid derivative of the formula I represented by the formula 4, wherein R is H.

Example 2: Analysis of inhibition of replication of hepatitis B virus gene

Cell line culture: In the embodiment of the present invention, a liver cancer cell line HepG2.2.15 (provided by Academician Chen Peizhe of the Hepatitis Research Center of National Taiwan University Hospital) is used, and the liver cancer cell of the strain can naturally produce hepatitis B virus particles and release into the culture medium, so The virus infection mode was used as an activity test evaluation of the pharmaceutical composition of the present invention. The HepG2.2.15 cell line was cultured in MEM medium and contained 10% fetal bovine serum (FBS) but no antibiotics, and cultured in a 37 ° C, 5% carbon dioxide incubator. In addition, in the cytotoxic killing experiment part of the present invention, the hepatoma cell line HepG2 was used as a control group for the cell survival experiment, and the HepG2 cell line was cultured by adding 10% FBS, 100 active units/ml (U/ml) penicillin (penicillin). ) Incubate with RPMI 1640 medium of 100 μg/ml (μg/ml) streptomycin at 37 ° C in a 5% carbon dioxide incubator.

Hepatitis B virus gene replication assay experiment: 5×10 ⁶ HepG2.2.15 cells were cultured in a 10 cm culture dish at different concentrations (1 μg/ml, 5 μg/ml, and 10) of the pharmaceutical composition of the present invention. Micrograms/ml) for 9 days; or after treatment at different concentrations (10 to 9 days) at a concentration of 10 μg/ml, remove the culture solution and add 1 ml of cell lysis buffer {component: 10 millimolar (mM) Tris hydrochloride (pH = 7.9), 1 mM concentration of edetic acid (EDTA), 0.1% sucrose solution [containing 50 mM ethyl phenyl polyethylene glycol ( Nonidet P-40) and 8% sodium chloride (NaCl)]} were lysed, centrifuged at 1300 rpm for 2 minutes and the supernatant was collected. The supernatant was added to Magnesium acetate until it was The final concentration was 6 millimolar and was reacted for 15 minutes at 37 ° C by the addition of 100 μg/ml DNase I. After the reaction was completed, 6.5% polyethylene glycol solution (Polyethylene glycol), 0.35 molar sodium chloride, and 10 mM concentration of edetic acid were added, and reacted at 4 ° C for 30 minutes, at 1300 rpm. Centrifuge for 4 minutes at a speed to collect the precipitate, and add the precipitate to 100 μl of trimethylolamine hydrochloride (pH=7.9) containing 10 mM, magnesium acetate at 100 μg and 100 μg/ml. Resuspend the pellet in DNase I buffer and react for 10 minutes at 37 ° C, then add 300 μl of sodium dodecyl sulfate-pronase cell lysis buffer ( 25 mM concentration of tris (hydroxyl aminomethane hydrochloride) [pH = 7.5], 10 mM concentration of edetic acid, 0.1 molar sodium chloride, 0.5% sodium lauryl sulfate) at 37 ° C After reacting for 30 minutes, an equal volume of phenol was added after the reaction was completed. After mixing and centrifuging, the aqueous layer liquid was collected, and the collected liquid was added to alcohol to precipitate hepatitis B virus DNA. After centrifugation, the precipitate was separated into hepatitis B virus DNA. .

Electrophoresis was carried out with 1% agar and 1X TAE buffer, and then transferred to a nitrocellulose membrane (NC membrane, Amersham Biosciences UK) for Southern blot analysis, and the hepatitis B virus DNA transferred to the nitrocellulose membrane was ^32. After the P-labeled hepatitis B virus DNA probe is hybridized, the nitrocellulose membrane is washed and exposed using a radioactive film (X-ray film), and the amount of viral DNA present in the film is scanned by an instrument. Quantitative analysis.

Southern blot analysis of viral DNA showed that HepG2 2.2.15 cells were treated with different doses of the pharmaceutical composition for 9 days, and the treatment dose was 10 μg/ml, which significantly inhibited the performance of viral DNA, and The effect of increasing the concentration of the pharmaceutical composition treatment and inhibiting the expression of viral DNA is more pronounced (Figure 5). In addition, treatment of HepG2 2.2.15 cells at a dose of 10 μg/ml for 6 to 9 days showed that the effect of the pharmaceutical composition on inhibiting viral DNA replication was more pronounced with increasing treatment time (Figure 6).

Example 3: Analysis of the effect of inhibiting the expression of hepatitis B virus surface antigen (HBsAg)

The HepG2 2.2.15 cell strain was purified by the pharmaceutical composition of the present invention at a concentration of 10 μg/ml for 6 days and 9 days, and the protein of HepG2 2.2.15 cell strain was purified, and after protein electrophoresis, it was transferred to polyvinylidene fluoride. Membrane (PVDF membrane, Amershan biosciences, UK) above, 4% skim milk powder solution (solvent is PBST buffer, containing 0.05% Tween-20 [Tween-20] phosphate buffer solution [PBS]) at room temperature After soaking for 1 hour, the virus surface antigen protein antibody serum (anti-preS, anti-preS2 or anti-major S antibody) was reacted for 1 hour, and then the polyvinylidene fluoride membrane was washed 3 times with PBST buffer (5 times each time). Minutes, and then reacted with a secondary antibody such as donkey anti-rabbit IgG or sheep anti-mouse IgG (Amershan Biosciences, UK) for 40 minutes, then washed 5 times with PBST buffer. (5 minutes each time), using an enhanced chemical cold detection kit (ECL detection kit) for color reaction, exposure using a radioactive negative (X-ray film), and then using a chemiluminescence image analyzer (chemiluminescence image analyzer, FLA 1000 system; Fuji Photo Film, Tokyo, Japan) Analysis of expression levels of hepatitis B virus surface antigen (HBsAg) a.

The experimental results showed that the expression level of the hepatitis B virus surface antigen (HBsAg) was significantly decreased after the HepG2 2.2.15 cell strain was treated by the pharmaceutical composition of the present invention. Moreover, the time for treatment of the pharmaceutical composition is increased, and the situation in which the performance of the hepatitis B virus surface antigen (HBsAg) is inhibited is more remarkable (Fig. 7).

Example 4: Analysis of inhibition of hepatitis B virus surface antigen (HBsAg) and e antigen (HBeAg) secretion

The inhibition of hepatitis B virus surface antigen (HBsAg) and e antigen (HBeAg) secretion assays was analyzed using radioimmunoassay (RIA). The HepG2 2.2.15 cell line was collected for 6 days and 9 days at a concentration of 10 μg/ml after the pharmaceutical composition of the present invention, and the culture supernatants were collected for radioimmunoassay kit (RIA). Assay kit, General Biologicals Corp., Industrial Park, Hsin Chu, Taiwan) was analyzed for quantification.

The experimental results showed that the hepatitis B virus surface antigen (HBsAg) and e antigen (HBeAg) proteins secreted into the culture medium by HepG2 2.2.15 were significantly decreased by the treatment with the pharmaceutical composition, and the treatment time was increased with the treatment time of the pharmaceutical composition. The inhibition is more pronounced, with up to 93% inhibition activity. In particular, the secretion of hepatitis B virus surface antigen (HBsAg) showed a significant inhibitory effect on the sixth day of treatment of the pharmaceutical composition (Fig. 8).

Example 5: Analysis of the toxicity of liver cancer cells (MTS assay)

HepG2 2.2.15 liver cancer cell line (2×10 ⁵ cells/well [cell/well]) was cultured in a 96-well culture plate (with HepG2 cell line as an experimental control group). After 24 hours, the medium was changed to contain different concentrations. Pharmaceutical composition MEM medium (0 μg/ml, 0.625 μg/ml, 1.25 μg/ml, 2.5 μg/ml, 5 μg/ml, 10 μg/ml, 20 μg/ml and 40 μg/ml) (not The cells were treated with fetal calf serum for 3 days. After the treatment, the culture solution was removed, washed three times with PBS, and 200 ml/well (ml/well) of MEM medium (excluding fetal bovine serum) and 20 ml/well were added. 2.5 mg/ml tetrazolium nitrogen compound ((3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2 a solution of hydrogen tetrazolium compound, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) Promega, Madison, WI), The reaction was carried out in a dark room at 37 ° C for 3 to 4 hours, and then the value was measured by a spectrophotometer at the absorption wavelength of OD490, and the cell growth inhibition ratio was further calculated by the following formula:

The results of the experiment showed that HepG2 2.2.15 liver cancer cells were treated with the concentration of the pharmaceutical composition of the present invention more than 10 μg/ml (20 μg/ml, 40 μg/ml), and the growth of the liver cancer cells was significantly inhibited, and the treatment concentration increased. The cytotoxic effect is more obvious, especially when the concentration of the pharmaceutical composition of the invention is 40 μg/ml, the cell killing effect can be as high as 90% (Fig. 9).

Scheme 1 is a flow chart for the preparation of a pharmaceutical composition.

Figure 2 is a ¹ H-NMR analysis spectrum showing the structural analysis of linoleic acid compounds.

Figure 3 is a ¹³ C-NMR analysis spectrum showing the structural analysis of linoleic acid compounds.

Figure 4 is a structural formula of a linoleic acid compound.

Scheme 5 shows the results of inhibition of hepatitis B virus DNA replication by treatment concentrations of the pharmaceutical compositions of the invention at various concentrations.

Scheme 6 shows the results of inhibition of hepatitis B virus DNA replication by different pharmaceutical composition treatment times.

Scheme 7 shows the results of inhibition of hepatitis B virus surface antigen (HBsAg) protein performance by different pharmaceutical composition treatment times.

Scheme 8 shows the inhibition of hepatitis B virus surface antigen (HBsAg) and e antigen (HBeAg) secretion results by different pharmaceutical composition treatment times.

Scheme 9 shows that the pharmaceutical compositions of the present invention at different concentrations inhibit the growth of liver cancer cells.

Claims (9)

Use of a linoleic acid compound of the formula I for the preparation of a medicament for the treatment of hepatitis B, Wherein R is H. The use according to claim 1, wherein the linoleic acid compound is obtained by extracting from a Solanaceae plant ( Solanaceae ). The use of the invention of claim 2, wherein the Solanum erianthum is Solanum erianthum . The use of claim 1, wherein the medicament further comprises a pharmaceutically acceptable carrier. Use of a linoleic acid compound of the formula I for the preparation of a medicament for the treatment of cancer, Wherein R is H. The use according to claim 6, wherein the linoleic acid compound is extracted from a Solanaceae plant ( Solanaceae ). The use according to claim 7, wherein the Solanum erianthum is Solanum erianthum . The use of claim 6, wherein the medicament further comprises a pharmaceutically acceptable carrier. The use of the invention of claim 6, wherein the cancer is liver cancer.