TW201016214A - Synergistic pharmaceutical composition for the treatment of cancers - Google Patents

Abstract

The present invention relates to a synergistic pharmaceutical composition for treating cancers, comprising synergistic effective amounts of isochaihulactone and an anticancer agent selected from the group consisting of an antimitotic drug and a topoisomerase I inhibitor, wherein the combination index of the pharmaceutical composition is less than 1. The pharmaceutical composition is a useful anticancer medicament, particularly for the treatment of human solid cancers, such as lung cancers, liver cancers, brain tumors, ovarian cancers, and colorectal cancers.

Description

201016214 IX. Description of the Invention: [Technical Field] The present invention relates to a pharmaceutical composition for treating cancer, and in particular to a method comprising isochahaihu (actushai) and a specific anticancer agent A synergistic pharmaceutical composition that enhances the treatment of human parenchymal cancers such as lung cancer, liver cancer, brain tumors, ovarian cancer, and colorectal cancer without increasing toxicity. _ [Prior Art] 瘳 cancer, a cell proliferative disease, has become the top of the top ten causes of death. In recent years, the top ten cancer deaths in men have been the first to replace lung cancer with liver cancer, the second is lung cancer, and female lung cancer The first person who is the leading cause of cancer death. Liver cancer has been the leading cause of cancer death in men for many years. Each year, the number of deaths due to liver cancer is as many as four or five thousand, and the age of death is concentrated between 4 and 6 years old, which is a period of life and a pillar of the family. Early liver cancer usually does not produce sputum symptoms, and when there are symptoms such as upper abdominal pain, loss of appetite, mild jaundice, general fatigue, weight loss, nausea, ascites and hepatomegaly, it is usually advanced liver cancer, but late The survival time of liver cancer is only 6 to 9 months. Therefore, only early diagnosis and timely medical treatment can have a chance of long-term survival. According to the pathological classification of the World Health Organization, lung cancer is divided into two categories: small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC). Compared with small cell lung cancer, the growth of non-small cell lung cancer is slower, and the metastasis is also worse than that of 悍 悍 , 129 129 129 129 129 129 129 129 129 129 129 129 129 129 129 129 129 129 129 129 129 129 129 129 129 129 129 129 129 129 129 129 129 129 129 The treated cases are only about 15 to 18%, and the chance of metastasis or recurrence after surgery is very high, and cases that cannot be operated are mostly insensitive to chemical drugs and radiation therapy. For the above reasons, the prognosis of lung cancer patients is not good. Overall, the five-year survival rate was only about 10 〇/〇 (National Institutes of Health '1998). • The treatment of cancer can be performed in several modes, including surgery, radiotherapy, chemotherapy, or a combination of the above. # For the treatment of lung cancer 'From the beginning of the year, the most common use is the beginning of chemotherapy drugs, such as cisplatin, the life expectancy of patients can be extended by one to two months." Since 1995, several new generations of chemotherapy The market for drugs has a good effect on the treatment of non-small lung cancer, including Pacific purple (4) (pacmaxel), European paclitaxel (d〇cetaxei), vinblastine. (VinblaStine), vincristine, vinorelbine (vinorelbine) and irinotecan (irin〇tecan, also known as cpTu). Pacific t-alcohol is mainly used in the treatment of cancers such as ovarian cancer, metastatic breast cancer, and lung ginseng melanoma. The mechanism of action of killing tumor cells is by the ratio of paclitaxel mi and binding to the cytoskeleton (cyt〇skelet〇n). $Tubulin (β-tubulm) to inhibit the depolymerization of microtubules (micr〇Ujbu(4)' thereby blocking mitosis, & inhibiting tumor growth, and inducing tumor cells> Zhou died (8) af0skl0nny et al., 1995). Therefore, treatment of early ovarian cancer with taXaneS is usually effective in killing tumor cells, which increases the patient's two-year survival rate by about 15. /(^Although Taipingol is already the second-line clinical use, but about 15 to 2 will develop anaphylactic shock. In addition, neurotoxicity is the clinical main; = 129356.doc 201016214. Vinca base compound Including vinblastine, vincristine, and vinorelbine, in contrast to the action of paclitaxel, it inhibits the polymerization of microtubules, thereby inhibiting the formation of spine during mitosis of cancer cells. 'National Breast Cancer and Colorectal Cancer Auxiliary Treatment Plan"

The National Surgical Adjuvant Breast and Bowel Project; NSABP) first showed that stage n and stage (1) colon cancer patients were treated with M0F chemotherapy (composed of 5-FU, methyl-CCNU and vincristine). ), can improve recurrence-free survival rate, and improve the overall survival rate of about 8% in five years. However, such drugs can also cause side effects such as neurotoxicity and bone marrow suppression. Irinotecan is metabolized to the active metabolite 7_ethyl_丨〇_hydroxy-Xishu (SN 38)' and combined with the 疋-isomerase (t〇p〇is〇rnerase I) to induce single-strand Defects in DNA production prevent DNA from replicating, causing cell death and shrinking the tumor. It has been clinically proven to have significant antitumor activity against several solid tumors with little or no sensitivity to chemotherapy, such as non-small cell lung cancer, cervical cancer, and colorectal cancer. In addition, it has considerable therapeutic effects on other tumors with chemosensitivity, such as ovarian cancer and small cell lung cancer. However, diarrhea caused by destruction of the gastrointestinal tract is the main clinical side effect. The above anticancer drugs are still used continuously for the treatment of patients with sputum cancer, but the prognosis of non-small cell lung cancer in the case of distant metastasis is not good, even if it includes the inclusion of cis-turned and combined guitar gemcitabine, vinorelbine Or combined chemotherapy of Pacific yew drugs, the vast majority of patients will still die 129356.doc 201016214 within a year, five-year survival rate. It is known that drugs are less than 10/° in the field of anticancer. Therefore, in view of the aforementioned safe anticancer drugs, sputum, efficacy needs to be searched for - more effective and [invention] to meet the demand for treating cancer diseases. The inventors of the present application found that the cleavage drug; the 5-phase and the lining ** '*day/, an anticancer agent selected from the group consisting of anti-silkase 1 inhibitors:

It does not increase toxicity. The effect is to achieve an enhanced cancer therapeutic effect. Therefore, the present invention is to provide a method for treating cancer, for example, preferably lung cancer, liver cancer, brain tumor, ovarian pain, and sputum. A synergistic pharmaceutical composition for fruit cancer and colorectal cancer, which comprises an effective amount of iso-caprolactone and a composition selected from the group consisting of k-filament cleavage and aglycone-converting enzyme ice preparation. A group of anticancer agents, wherein the pharmaceutical composition has a combination index index, CI) of less than one. [Examples] Isocaprolactone used in the present invention belongs to lignin (Ugnan), which is a heterocyclic ring having a γ-butyrolactone structure as a core and a carbon 2_(5) in a 2 configuration or an E configuration. Compounds, which are disclosed in U.S. Patent Application No. 2005/0013879 A1 to U.S. Patent Application Serial No. 2006/0079575, the entire disclosure of which is incorporated herein by reference. -3-(3,4,5-trimethoxy-benzylidene)·dihydrofuran·2 ketone O-benzonAdioxoHylmethyl-Sp^J-trimethoxyl-benzylideneydihydro-furan-Z-one) with the following chemistry Structural formula: 129356.doc 201016214

Isocaprolactone has antitumor activity against non-small cell lung cancer and inhibits the growth of cell lines by causing apoptosis. This compound has no drug resistance to cell lines resistant to paclitaxel # (Cr〇SS reSiStanCe). It is administered subcutaneously, and it has obvious tumor suppressing effect on the ectopic transplantation mode of human malignant lung cancer cells in nude mice, and the biochemical values and histopathological sections after administration are observed, and there is no bone cancer inhibition or other organ damage. This may be due to the high specificity of the toxicity of isobutadione to human liver cancer, lung cancer, brain tumors, ovarian cancer and colorectal cancer. Isocaprolactone suitable for use in the present invention may comprise asymmetric centers, or racemates, racemate mixtures or various diastereomers, all isomeric forms of the compounds being included In the present invention. In the present invention, an anticancer agent which can be used in combination with isobutadione in an anticancer therapy can select a drug which is currently clinically useful and valuable, and includes an anti-mitotic drug or a site-directed invertase] [inhibition] Agent. The pharmaceutical composition thus prepared is an effective anticancer drug which can synergistically act to effectively inhibit tumor cell growth, has an enhanced antitumor effect, and can reduce side effects caused by antitumor. The term "anti-cancer therapy" as used herein refers to the treatment of human solid cancer 129356.doc -10· 201016214 ovarian cancer, large intestine straight, sputum cancer, kidney can also be based on solid tumor, including liver cancer A pharmaceutical composition of all types of therapy, lung cancer, brain tumor, intestinal cancer, melanoma, breast cancer, prostate cancer, gastric cancer, and central nervous system tumors is used in the above anticancer therapy. In a preferred embodiment of the present invention, The pharmaceutical composition can be used for the treatment of liver cancer, lung cancer, especially non-small cell lung cancer, including initial, clinical drug treatment and metastatic lung cancer, or paclitaxel-resistant lung cancer, or brain cancer. "Effect" means inhibiting tumor growth or completely resolving the tumor. The so-called "reducing the side effects of anti-tumor" means that there is no obvious physiological toxicity or bone marrow suppression, neurotoxicity and the like. * The anti-mitotic agent suitable for use in the present invention may be any anti-mitotic drug known in the art to be suitable for treating cancer, such as, but not limited to, paclitaxel, taxol, vinca, vinca, vinorelbine, Or a hydrate thereof, an isosteres and a pharmaceutically acceptable salt. The localization of the isomeric invertase j inhibitor suitable for use in the present invention may be any of the known stereoisomerase inhibitors known in the art to be useful for the treatment of cancer, such as, but not limited to, irinotecan' Topotecan (t〇p〇tecan), 7_ethyl-Guangxijixishu (SN_38), or a hydrate thereof, an electron-aligned body, and a pharmaceutically acceptable salt. In a specific embodiment of the present invention, the pharmaceutical composition preferably comprises an iso-pyrene and an isomerization-converting enzyme 丨 刽, plus i, ^ '

129356.doc 201016214 The pharmaceutical composition of the present invention, in addition to the active ingredient described above, may further comprise other components well known to those of ordinary skill in the art, such as, but not limited to, pharmaceutically acceptable. Carrier, excipient, buffer or preservative, or other related anticancer drug. The active ingredient of the pharmaceutical composition t of the present invention can be administered simultaneously, separately or in any order in a continuous manner, for example, by administering isobuta (4) and paclitaxel from day 1 to day 5.

The term "administration" or "administration" as used herein means that the active ingredient in a pharmaceutical composition can be administered to a patient by any means medically determined, for example, by oral administration, non-success, nasal or topical administration. The actual preferred method and sequence of administration should be adjusted according to the utilization of the particular formulation of the active ingredient, the tumor transplant model to be treated, and the individual differences of the patient. The form of the pharmaceutical composition of the present invention can be formulated as oral. Suspension tablets, troches, capsules, troches, emulsions, powders, solutions, etc. Parenteral administration includes administration of components via subcutaneous, intravenous or intramuscular injection, for example, _ sexual components are dissolved in physiologically acceptable diluents Injectable form, together with a pharmaceutically acceptable carrier such as water, physiological saline, aqueous dextrin or related sugar solution, ethanol, polyethylene glycol 4GG or oil, etc. The drug can also be made into an inhalant or a suspension drug. Absorbed through the skin or mucous membrane. In the specific embodiment of the present invention, the "during the field" by the soil injection method can surely control the time and dosage of administration, which is a preferred route of administration. The pharmaceutical composition of the present invention can be formulated by the method of the invention in the technical field or the improvement thereof, and is not particularly limited, and can be prepared according to the method of mixing, and the order of addition of each component can be (4) 2 129356.doc S26367 «9356 〇07〇〇η·ν£. •12- 201016214 In this launch, the compound is formulated as 'for example, the appropriate dose of Isoprofen can be pre-dissolved in a solvent, eg == (DMS咐With a suitable proportion of co-solvent, such as chlorinated E. Cremophor EL, diluted into a suitable dose via a diluent, such as physiological saline, into a subcutaneous injection.: - in the course of treatment, based on patient weight The dosage of Isophora vinegar is 自 自 ^ from about (M mg / kg to about 1000 mg / kg, the anti-silk 2 music is from about 05.05 mg / kg to about 500 mg per day) The administration of the ortho-inhibitor 1 inhibitor is from about 05 mg/kg to about 5 mg/kg per day, which is effective for the treatment of the above diseases. The dosage can be administered according to various administration methods. The knowledge and technology known in the art changes on its own 'for example' The oral administration method should be increased by two to five times. In addition, it is known in the art that the content of the active ingredient and the frequency of the drug in the pharmaceutical composition will vary according to the severity and individual differences of the patients treated by the article, such as the active ingredient. Factors such as metabolic stability and length of action, patient age, body weight, health status, sex, diet, route of administration, time of administration, rate of excretion, and excipients, etc., which may be common to those skilled in the art Decide. - The preferred embodiment of the month of the month includes the combination of iso-Bupiva vinegar and anti-mitotic drugs, the use of sigma and the combination of iso-invertase I inhibitor and the isomerase I inhibitor. In the invention, the concentration ratio of iso-Bupiva vinegar to the anti-mitotic drug is about 30,000 : s to about 100: 1; and the concentration ratio of iso-caprolactone and the positional isomerase 1 inhibitor is about no: i to about 10: 129 356.doc 201016214 The following examples are intended to further illustrate the invention, but are not intended to limit the scope of the invention. Modifications and variations that may be readily made by those skilled in the art are included within the scope of the disclosure of the present disclosure and the scope of the appended claims. EXAMPLES The following examples describe the ability of the pharmaceutical compositions of the present invention to inhibit the growth of lung cancer, liver cancer and brain tumor cells by a known technique, and the results, and the methods and steps involved in the analysis method are all in the technical field. Among those who are known to the general knowledge. The present invention was carried out to treat the A549 lung cancer cell line (cell Une), the j5 liver cancer cell line, the DBTGR-05 MG brain tumor cell line, and the nude mouse subcutaneous tumor cell growth inhibition as an effective model for drug utility, to study the present invention. The effectiveness of the pharmaceutical composition. MTT drug combination analysis 医药 The pharmaceutical composition was studied using a Μττ cell survival assay (Pavwels et al., J. Virol. Methods, 1988' 20, 309-321). The method of adding the active ingredient in the pharmaceutical composition is to drip the iso-Bupouin g along the inner wall of the 96-well (we 11) microplate, and then continuously drip the anticancer drug, and use 4 microplates for each analysis. Four wells are used for each combination of drugs. The drug was diluted twice or half, and the first was discharged into the solvent as a control group. Usually, 3x103 A549 lung cancer cells were cultured in each well, cultured until the next day, and the drugs were added alone or in different combinations for 2 days, and then the culture solution was aspirated and then added to a medium containing 500 ng (μβ)/ml of MTT. The culture medium of I29356.doc -14· 201016214 (μΐ) was further cultured for 4 hours, and the culture solution was aspirated and then added with 2 μM microliters of dimethyl hydrazine, and then measured at 57 〇N by a microplate spectrophotometer ( The absorbance of nm). The mean 〇D of each drug combination can be calculated as an inverse of the different drug doses. The readings of the highest dose wells were used as the positive control group (the growth of living cells was not inhibited), and the first row of solvent-only wells was read as the negative control group. Thus, the iC50 of the drug (concentration for inhibiting the growth of half of the cells) can be obtained. The survival rate of the 'negative control group was 1 〇〇 0 / 〇, and the lethal rate was (1 〇〇 / / - survival rate = lethal rate). ❹

CalcuSyn software analysis of the synergistic effect of pharmaceutical compositions In this application, the CalcuSyn software is used to draw the drug dose response curve, and the IC5 of each drug combination, whether alone or at a fixed concentration, is the data of other drug combinations. Calculated and recorded on the equal weight map. CalcuSyn software is mainly used for the analysis of synergy or antagonism in the treatment of cancer or AIDS, which can perform the dose-effect calculation of a variety of drugs, and this algorithm is based on T_c 〇Chou Developed jointly with Ρ· Talalay et al. First, we need to calculate the concentration of each drug in the IC50 of each cancer cell. The concentration is defined as 1, plus 5 concentrations of 〇·25, 0.5, 2, and 4 times of this concentration are added together. The cancer cells were analyzed by MTT drug combination, and then one of the drugs was halved by the above five concentrations, and then the repeated combination of Μττ drugs was analyzed, then the mortality rate was calculated, three curves were observed, and the combination index of the pharmaceutical composition was calculated. It can be determined whether the drug has a synergistic relationship, an additive effect or an antagonistic effect. (Chou, TC and Talalay, P. Quantitative anaiysis of dose_effect relati〇nships: eg 129356.doc -15· 201016214 combined effects of multiple drugs or enzyme inhibitors. Adv. Enzyme Regul·, 22:27-55, 1984; Chou, TC The median-effect principle and the combination index for quantitation of synergism and antagonism. In: TC Chou and DC Rideout (eds.), Synergism and Antagonism in Chemotherapy, pp. 61-102. New York: Academic Press, 1991; Chou, J. Quantitation of synergism and antagonism * of two or more drugs by computerized analysis. In: TC

Chou and D. C. Rideout (eds.), Synergism and Antagonism in Chemotherapy, pp. 223-244. New York: Academic Press, 1991). ' The combination index (Cl) can be determined from these data and the following formula: CI^ (D), , (P)2, α(Ρ), (Ρ)2 (DX)1 (DX)2 (DX), (DX 2) The α-compatible reagent is 1 φ (DX)i = IC50 of the above drug 1 (DX) 2 = IC50 of the above drug 2 (Dh & (D) 2 = 50% after drug 1 and drug 2 combination If the concentration at the time of inhibition is <1, the synergistic CI = 1 has no effect on each other> 1 is mutually antagonistic (Reference: J. Suhnel, Antiviral Research 13 (1990), 23-40) Example 1 129356. Doc -16- 201016214 Anticancer activity of paclitaxel, vincristine and irinotecan. Three anticancer agents, Pacific Paclitaxel (Tables 1A and 1B), vincristine (Tables 2A and 2B), and irinotecan (Tables 2A and 2B) Tables 3A and 3B) and Isocaprolactone (K8) are studied in the same combination. The method for testing the effectiveness of the composition is to test the antitumor cell activity of different concentrations of the drug combination using the aforementioned cell survival assay. The method can determine whether there is synergy by analyzing the combination index of the data. A549 lung cancer cell line® After 48 hours of administration to A549 cell line, the IC50 of each of the following active ingredients was determined: ΙΟ50(μΜ) Isoflurane 40 Pacific Paclitaxel 0.005 Changchun New Test 0.005 Irinotecan 500 Tables 1A and 1B are the equivalent doses of Isocaprolactone (ie Isocaprolactone: . Pacific Paclitaxel) =4000:1) and the results of the MTT test of the half dose of Pacific paclitaxel (ie isobutadione: Pacific paclitaxel = 16000:1), the CI values were 0.3 and 0.87, respectively, indicating that the two combinations have synergy Sex.

Table 1A Isocaprolactone (μΜ) Pacific paclitaxel (ηΜ) Mortality rate (%) 80 20 67.3 40 10 67.2 20 5 48.8 10 2.5 39 5 1.25 31.7 129356.doc -17· 201016214 CI=0.3, indicating Synergy

Table 1B Isocaprolactone (μΜ) Pacific paclitaxel (ηΜ) Mortality rate (%) 160 10 79.4 80 5 73.9 40 2.5 63.2 20 1.25 40.8 10 0.625 34.9 CI=0.87, indicating synergy

Tables 2A and 2B show the dipyridamole dose half (ie iso-caprolactone: vincristine = 4000:1) and vincristine dose half (ie iso-caprolactone: vincristine = 16000). The MTT results of :1) showed CI values of 0.64 and 34.6, respectively, indicating that the combination of isobutadione: vincristine = 4000:1 was synergistic.

Table 2A Isocaprolactone (μΜ) Vincristine (ηΜ) Mortality rate (%) 80 20 74.2 40 10 61.5 20 5 52.4 10 2.5 48.3 5 1.25 45.9 CI=0.64, indicating synergy

Table 2B Isocaprolactone (μΜ) Changchun New Test (ηΜ) Mortality rate (%) 160 10 74.4 80 5 68.5 40 2.5 57.0 20 1.25 55.8 10 0.625 66.9 129356.doc -18· 201016214 CI=34.6, indicating that it does not have Synergy Tables 3 A and 3B are the half doses of isobutadiol (ie, Isobutanol: irinotecan = 40:1) and the dose of irinotecan, respectively (ie, Isocaprolactone: Iritidine The MTT results of Kang = 160:1) showed CI values of 0.07 and 3.23, respectively, indicating that isobutadione: irinotecan = 40:1 combination was synergistic.

Table 3A Isocaprolactone (μΜ) Irinotecan (μΜ) Mortality rate (%) 80 2 60.4 40 1 56.0 20 0.5 55.3 10 0.25 62.2 5 0.125 76.3 CI=0.07, indicating synergy

Table 3B Isocaprolactone (μΜ) Irinotecan (μΜ) Mortality rate (%) 160 1 63.9 80 0.5 60 40 0.25 55.6 20 0.125 55.8 10 0.0625 48.8 CI=3.23, indicating that there is no synergistic J5 旰 cancer cell line After 48 hours of dosing to J5 hepatoma cell line, the IC50 of each of the following active ingredients was determined: 129356.doc -19- 201016214 Active ingredient IC50 (_ Isocaprolactone 20 Pacific paclitaxel 0.02 Changchun Xinzheng 0.01 Ilitico 1000 Table 4 A and 4B are the equivalent doses of isobutadione (ie iso-caprolactone: too citrusol = 500:1) and the dose of Pacific paclitaxel are half (ie iso-caprolactone) : The results of MTT test of Pacific paclitaxel = 2000:1), the CI values of φ are 0.22 and 0.06, respectively, indicating that the two combinations are synergistic.

Table 4A Isocaprolactone (μΜ) Pacific paclitaxel (ηΜ) Mortality rate (%) 40 80 71.2 20 40 69.5 10 20 64 5 10 60.6 2.5 5 53.2 CI = 0.22, indicating synergy ❿ Table 4 Β Different firewood Hu lactone (μΜ) Pacific paclitaxel (ηΜ) Mortality rate (%) 80 40 70 40 20 67.8 20 10 60.9 10 5 58.3 5 2.5 53.3 CI=0.06, indicating synergistic DBTGR-05MG brain tumor cell line dosing After 48 hours to DBTGR-05MG brain tumor cell line, IC50 of the following active ingredients of 129356.doc -20· 201016214 was determined: active ingredient IC50 (_ isocaprylactone 50 pacificol 20 vincent new test 2 irilide Kang 4000

Tables 5A and 5B are the dipyridamole doses in half (ie, isobutadiol: Pacific paclitaxel = 1250:1) and the paclitaxel dose in half (ie isobutadione: Pacific purple shirt) The MTT results of the alcohol = 10000:1), the CI values were 0.22 and 0.06, respectively, indicating that the two combinations are synergistic.

Table 5A Isocaprolactone (μΜ) Pacific paclitaxel (uM) Lethality (%) 100 80 76.2 50 40 74.1 25 20 62.3 12.5 10 64.2 6.25 5 56.6 CI=0.3, indicating synergy

Table 5B Isocaprolactone (μΜ) Pacific paclitaxel (uM) Lethality (%) 200 40 78.8 100 20 70.6 50 10 70.2 25 5 66.2 12.5 2.5 65.2 CI=0.06, indicating synergy. Example 2 129356.doc -21 - 201016214 Intra-pacific and anti-cancer activity of paclitaxel and iso-Bupivain g|Paclitaxel is an anti-mitotic chemotherapeutic drug, which is different from eucalyptus Together, they are effective representative pharmaceutical compositions. The method for testing the effectiveness of the pharmaceutical composition is to observe the in vivo antitumor activity of the pharmaceutical composition by subcutaneous tumor transplantation mode. It is mainly observed whether the lower drug combination dose achieves the same tumor suppressing effect as the drug alone. Table 6 shows the activity analysis of the combination of Pacific paclitaxel and isocaprolactone against subcutaneous A549 lung cancer in nude mice. — Time of drug treatment (1,2) Dose (mg/kg/day) Relative tumor size (multiple) (3) Statistical analysis (4) Solvent iP for 5 consecutive days 0 6.31 Different Bupleurum ip for 5 consecutive days 10 2.74 Ρ=0.30~ Pacific paclitaxel iP·5 days in a row 5 3.39 Ρ=0.31 Echinacea S-same + 10 Pacific paclitaxel------- ι·Ρ. 5 days in a row 50 2.25 Ρ=0.30 ❹ (1) In vivo culture of 5xl〇6 A549 lung cancer cells was injected into Baib/c immunocompromised nude mice for in vivo tumor transplantation experiments. (2) After the tumor grows to 1〇〇mm3 (长父宽乂宽/2), the subcutaneous (ι.Ρ·) administration is started, and the drug is administered more than 15 cm on the opposite side of the tumor, once a day for 5 consecutive days. . The combined treatment group was first administered with Isocaprolactone, and two hours later, Pacific Paclitaxel was administered. (3) On day 5, after continuous administration for 28 days, tumor size and body weight were measured every 3 days. The tumor size was measured on day 28 of each nude mouse divided by the number of days obtained on day 1 as the relative tumor size. . In this experiment, the tumor value 129356.doc -22· 201016214 was used as the evaluation result of the activity of inhibiting tumor growth, and the smaller the value, the better the inhibition effect. (4) Observing whether the relative size of tumors after treatment alone or in combination was significantly different from that of the control group (solvent only). P value less than 〇.5 indicates significant difference. Table 6 data clearly shows that compared with single administration, Pacific The combination of the taxol and the different 'Bupleurum vinegar can inhibit the growth of tumor cells more effectively and enhance the anti-tumor effect. ❹ Figure 1 shows the synergistic anticancer effect of using Isocaprolactone (K8) and paclitaxel to treat Α549 human lung cancer cells. Figures 2 and 28 show that the combined anti-cancer effect of Isoprolactone and paclitaxel in the treatment of Α549 human 'lung cancer cells may be through Phosphorus-ERK1/2 egg-white (PhosPhor-ERK1/2) and Split caspase (cleaved

A large amount of performance of CaSpaSe-3) (Fig. 2A) and NAG-1 protein (Fig. 2B). While the present invention has been described in its preferred embodiments, the present invention is not intended to limit the invention, and it is to be understood that those skilled in the art can make some modifications and refinements without departing from the spirit and scope of the invention. The scope of the invention is defined by the scope of the appended claims. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows the synergistic anticancer effect of the combination of isobutadione (K8) and paclitaxel in the treatment of Α549 human lung cancer cells. Figure 2Α and 2Β show that the combined anti-cancer effect of Isoflurane and paclitaxel in the treatment of Α549 human lung cancer cells may be through the large-scale performance of phosphorus-ERK1/2 protein (phosphor_ERK1/2) and then 129356 .doc •23· 201016214 Cleaved caspase-3 (Fig. 2A) or a large amount of performance through the NAG-1 protein, which in turn causes cell growth inhibition (Fig. 2B).

129356.doc -24-

Claims (1)

201016214 X. Patent Application Range: 1. A pharmaceutical composition for treating cancer comprising a synergistically effective amount of isochaihulactone and consisting of an anti-mitotic drug and a localization-isomerase I inhibitor A group of anticancer agents, wherein the pharmaceutical composition has a combination index of less than one. 2. The pharmaceutical composition according to claim 2, wherein the anti-mitotic drug is selected from the group consisting of paclitaxel 'docetaxel', vinblastine, vincristine, vinorelbine (vinorelbine), a group of hydrates thereof, an electron alignment thereof, and pharmaceutically acceptable salts thereof. 3. The pharmaceutical composition of claim 2, wherein the concentration ratio of isobutadione to the anti-filament split is from about 30,000:1 to about 100:1. 4. The pharmaceutical composition of claim 3, wherein the anti-mitotic drug is pacific paclitaxel. 5. The pharmaceutical composition of claim 4, wherein the isobutadione and the Pacific purple The concentration ratio of cedarol is about 4 〇 00:1, and the combined index of the pharmaceutical composition is equal to or less than about 0.3. 6. The pharmaceutical composition according to claim 4, wherein the concentration ratio of isobutadione to paclitaxel is about 16 〇〇〇:1, and the combination index of the pharmaceutical composition is equal to or less than about 787. The pharmaceutical composition of claim 4, the degree of incidence of cedarol is about 5: i, equal to or less than about 0.22. Wherein the combination of isobutanol and Pacific purple and the pharmaceutical composition is 8. The pharmaceutical composition of claim 4 wherein the ratio of the concentration of Isocarpus to the Pacific Violet 129356.doc 201016214 is about 2000: 1, and the combination index of the pharmaceutical composition is equal to or less than about 0.06. 9. The pharmaceutical composition according to claim 4, wherein the concentration ratio of isobutadione to paclitaxel is about 1250:1, and the combination index of the pharmaceutical composition is equal to or less than about 0.22. 10. The pharmaceutical composition of claim 4, wherein the concentration ratio of isobutadione to paclitaxel is about 1 :1, and the combined index of the pharmaceutical composition is equal to or less than about 0.06. 11. The pharmaceutical composition of claim 2, wherein the anti-mitotic drug is a new test of Changchun. A pharmaceutical composition according to claim 11, wherein the ratio of isobutadione to vincristine is about 4000:1, and the combination index of the pharmaceutical composition is equal to or less than about 0.64. The pharmaceutical composition according to claim 1, wherein the localization isomerase indole inhibitor is selected from the group consisting of irinotecan (irin〇tecan), topotecan (t〇p〇tecan), 7-acetamidine 4 A group consisting of a hydrocarbyl eucalyptus (SN-38), a # hydrate, an isoelectronic alignment thereof, and a pharmaceutically acceptable salt thereof. 14. The pharmaceutical composition of claim 13, wherein the concentration ratio of isobutanol to the localized isoformase 1 inhibitor is from about 150:1 to about 丨〇:丨. 15. The pharmaceutical composition of claim 14, wherein the localization isomerase! inhibitor is irinotecan. 16' The pharmaceutical composition of claim 15 wherein the ratio of isobutadione to irinotecan is about 40:1 and the combination index of the pharmaceutical composition is equal to or less than 129356.doc 201016214 17. The pharmaceutical composition of item 1, which further comprises a pharmaceutically acceptable carrier or excipient. The pharmaceutical composition of claim 1, wherein the isobutadione and the anticancer agent are administered simultaneously, separately or continuously. The pharmaceutical composition according to claim 1, wherein the cancer comprises lung cancer, liver cancer, brain tumor, ovarian cancer, and colorectal cancer. 129356.doc S26367 129356 ΟΟ7929762-1