JP2012017294A - Usage of 5-substituted 4, 7-dimethoxy-1, 3-benzodioxol for production of therapeutic medicine for colorectal cancer - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a cell cycle inhibitor, a medicine which kills colorectal cancer cells, and a production method of the medicine.SOLUTION: There is disclosed a 5-substituted 4, 7-dimethoxy-1, 3-benzodioxol expressed by formula (I). In the formula, R represents a substituted or non-substituted group selected from a group consisting of an alkyl group having 1 to 6 carbon atoms except for a methyl group, an alkenyl group having 2 to 6 carbon atoms, an alkynyl group having 2 to 6 carbon atoms, a cyano group, an ester group having 1 to 5 carbon atoms, a carboxy group having 1 to 5 carbon atoms and an isoxazole group.

Description

  The present invention relates to the manufacture of a medicament for the treatment of 5-substituted 4,7-dimethoxy-1,3-benzodioxole, a colorectal cancer treatment of 5-substituted 4,7-dimethoxy-1,3-benzodioxole. The present invention relates to a method for producing a pharmaceutical product for treating colorectal cancer. However, the 5-position substituent in 5-substituted 4,7-dimethoxy-1,3-benzodioxole does not include a methyl group. The present invention also relates to a cell cycle inhibitor and a colorectal cancer cell killing agent containing 5-substituted 4,7-dimethoxy-1,3-benzodioxole.

  According to a cancer report published by the Taiwan Health Department in 2006, colorectal cancer is one of the top 5 most common cancers in Taiwan. In current clinical treatment, treatment is limited to surgical treatment, radiation therapy, general chemotherapy, and gene therapy (Asmis TR. And Saltz L., Gastroenterol Clin North Am, 2008; 37: 287-95, ix Durai R. et al., J Gastrointest Liver Dis 2008; 17: 59-67; Kuwai T. et al., Clin Exp Metastasis, 2008; 25: 477-89). It is clear that there is still an urgent need for the development of new therapeutics used in the treatment of colorectal cancer.

  Antrodia camphorata is used in Chinese traditional medicine to treat hypertension, pruritus, diarrhea, or liver cancer. According to the applicant's previous report, 4,7-dimethoxy-5-methyl-1,3-benzodioxole (4,7-dimethyl-5-methyl-1,3-benzodioxole, hereinafter abbreviated as SY-1) It is isolated from beef turf extract. SY-1 can effectively suppress the proliferation response of human COLO 205 cancer cells (COLO 205) (Lien HM. Et al., Evid Based Complement Alter Med. Doi: 10.1093 / ecam / nep020, (Released on March 17, 2009).

  However, the anti-tumor activities of 4,7-dimethoxy-1,3-benzodioxole are not yet fully understood. Furthermore, although SY-1 has an effect of suppressing tumor growth, it can exert an effect of effectively suppressing colorectal cancer cells only at a relatively high dose. Therefore, there is an urgent need in the art for new drugs that can effectively treat colorectal cancer in order to increase the options for drugs used to treat colorectal cancer. In addition to SY-1 according to the prior art, if various candidate drugs for the treatment of colorectal cancer can be provided, by providing other options in advance, treatment can be performed when drug resistance occurs for a specific drug. It is possible to avoid the problem that there is no alternative medicine to be used for the treatment.

  In view of the various shortcomings and needs in the prior art, the present invention aims to provide the use of derivatives of compounds extracted from natural products in the treatment of colorectal cancer. The compound is 5-substituted 4,7-dimethoxy-1,3-benzodioxole and is provided in the form of a pharmaceutical composition containing a 4,7-dimethoxy-1,3-benzodioxole derivative. Can do.

  To achieve the above object, the present invention provides the use of 5-substituted 4,7-dimethoxy-1,3-benzodioxole in the manufacture of a medicament for the treatment of colorectal cancer, 7-Dimethoxy-1,3-benzodioxole has the following general formula (I):

In the above formula, R represents an alkyl group having 1 to 6 carbon atoms, an alkenyl group having 2 to 6 carbon atoms, an alkynyl group having 2 to 6 carbon atoms, a cyano group A group selected from the group consisting of a group having 1 to 5 carbon atoms, an ester group having 1 to 5 carbon atoms, a carboxyl group having 1 to 5 carbon atoms, and an isoxazole group. Yes, provided that (the the proviso that) R is not a methyl group (other than -CH3).

  The above groups may be substituted or nonsubstituted. More preferably, R is ethyl group, propyl group, vinyl group, allyl group, butadienyl group, ethynyl group, propynyl group, cyano group, methyl ester group, ethyl ester group and carboxymethyl group, carboxyethyl group or A group substituted or unsubstituted with a carboxylpropyl group.

  Preferably, R is the substituted functional group, and can include, but is not limited to: A substituted alkyl group having 1 to 6 carbon atoms, which is a hydroxy group (—OH), a cyano group, a nitro group, an aromatic group or a halogen group: substituted The substituted alkenyl group having 2 to 6 carbon atoms, the substituent is a hydroxy group, a cyano group, a nitro group, an aromatic group or a halogen group; the substituted alkynyl group having 2 to 6 carbon atoms, the substituent is A hydroxy group, a cyano group, a nitro group, an aromatic group or a halogen group;

R represents —CN, —COOH, —CH ₂ CH ₂ OH, —COOCH ₃ , — (CH ₂ ) ₂ CH ₃ , —CH ₂ CH (OH) CH ₂ CN, —CH ₂ CH═CH ₂ ,

and

Those selected from the group consisting of

Further, the half-inhibitory concentration (IC ₅₀ ) of the 5-substituted 4,7-dimethoxy-1,3-benzodioxole for the proliferation action of human colorectal cancer cells is preferably 375 μM or less, and is preferably 225 μM or less. Is more preferably 200 μM or less, and even more preferably 35 μM or less.

  Furthermore, the 5-substituted 4,7-dimethoxy 1,3-benzodioxole is apiole, ie 4,7-dimethoxy 5- (2-propylene-1-yl) -1,3-benzodiole. Xol is preferred.

  The present invention provides a pharmaceutical composition for use in the treatment of colorectal cancer, the pharmaceutical composition comprising: A therapeutically effective amount of any of the foregoing 5-substituted 4,7-dimethoxy-1,3-benzodioxole and enantiomers, non-enantiomers, pharmaceutically acceptable salts, or combinations thereof, and These pharmaceutically acceptable carriers and / or excipients.

  The said pharmaceutically acceptable salt means the salt produced | generated between the following pharmaceutically acceptable inorganic acids or organic acids. Examples include, but are not limited to, hydrochloric acid, sulfuric acid, sulfonic acid, acetic acid, propanoic acid, succinic acid, malonic acid, citric acid, tartaric acid, methanesulfonic acid, and p-toluenesulfonic acid.

  In the present invention, the above-mentioned “therapeutically effective amount” refers to administration of a therapeutically effective amount of a compound to a treatment subject. Evaluation as effective for treatment is judged objectively using a test or a marker or the like, or subjectively based on the treatment target or the treatment subject's sense for the sense after treatment. The effective amount range of the 5-substituted 4,7-dimethoxy-1,3-benzodioxole is from about 0.1 mg / Kg to about 500 mg / Kg, or from 1 mg / Kg to about 50 mg / Kg. . The effective amount may vary depending on the route of administration or other drugs used in parallel.

  5-Substituted 4,7-dimethoxy-1,3-benzodioxole of the present invention is an active ingredient in pharmaceutical compositions and can be administered orally or parenterally, intravenous, intramuscularly (Intramuscular), intraperitoneal, subcutaneous, rectal and topical routes of administration.

  For oral dosing, the 5-substituted 4,7-dimethoxy-1,3-benzodioxole of the present invention is combined with conventional antineoplastic and / or cell growth inhibitors. For example, in the form of tablets or capsules, powders, dispersible particles, flat capsules, aqueous solutions or suspensions. Commonly used carriers for oral administration in tablets are lactose, corn starch, magnesium carbonate, talc and sugar. Lubricants include, for example, magnesium stearate. When administered orally in capsule form, effective carriers are lactose, corn starch, magnesium carbonate, talc and sugar. For oral administration in an aqueous suspension, the carrier can be an emulsifier and / or a suspension. In addition, sweeteners and / or flavorings can be further contained during oral administration.

  For dosing via intramuscular, intraperitoneal, subcutaneous and intravenous, a commonly used dosage form is a sterile solution form of the active ingredient, the pH of which is adjusted by a buffer.

  A commonly used dosage form for intravenous administration is an isotonic solution obtained by controlling the overall concentration of the solute containing the active ingredient.

  For rectal administration, a commonly used dosage form is a suppository. A low melting point wax (for example, a mixture of fatty acid, glycerin or coconut oil) is dissolved in advance, then the active ingredient is dispersed in the low melting point wax by a method such as uniform stirring, and then the low melting point wax and the active ingredient are evenly distributed. The mixed mixture is poured into a mold of an appropriate size and cooled to form a solid.

  The pharmaceutical composition of the present invention can be used in combination with any known method or composition suitably used for the treatment of other colorectal cancers.

  The present invention provides a cell cycle inhibitor containing the 5-substituted 4,7-dimethoxy-1,3-benzodioxole.

  The present invention provides a colorectal cancer cell killing agent containing the 5-substituted 4,7-dimethoxy-1,3-benzodioxole.

  5-Substituted 4,7-dimethoxy-1,3-benzodioxole in the present invention has been shown to have specificity for cell type and kills colorectal cancer cells, but is normal. It does not kill human cells. Therefore, it effectively counters the proliferative action of colorectal lesion cells and can be used for the treatment of colorectal cancer. The 5-substituted 4,7-dimethoxy-1,3-benzodioxole and pharmaceutical compositions thereof of the present invention have a relatively low half-inhibitory concentration on the proliferative action of colorectal cancer cells. This indicates that it has the effect of effectively suppressing cancer cell growth at a relatively low dose compared to the prior art, and is a combinatorial combination of colorectal cancer cell killing drugs or other colorectal cancer therapeutics. Useful for Chemistry Data Bank.

It is a graph which shows a mode that cell growth is suppressed, after a human colorectal adenocarcinoma cell (COLO 205) was processed with SY-1 derivative for 24 hours. It is a graph which shows a mode that cell growth is suppressed, after a human colorectal adenocarcinoma cell (COLO 205) is processed by SY-1 derivative for 48 hours. It is a graph which shows a mode that cell growth is suppressed after a human colorectal adenocarcinoma cell (COLO 205) is treated with SY-1 derivative for 72 hours. It is a graph which shows a mode that a cell growth is suppressed after normal human cells (Normal human cells) (FHC cell) are processed for different time by SY-1 derivative of a different density | concentration. It is a figure which shows the dose dependence effect with respect to the stagnation of the G0 / G1 stage in COLO205 cell of apiol. This is obtained by flow cytometric analysis, the percentage of cells in the sub-G1, G0 / G1, S and G2 / M stages is measured by the Multicycle analysis software, each group has 3 samples, the values are averaged Values are expressed as ± SE and ^* P <0.05 relative to the control group. It is a graph which shows the effect with respect to the expression of a cell cycle adjustment protein of an apiol. Shows time- and dose-dependent responses of Apioles to apoptosis of COLO 205 cells, and after synchronized COLO 205 cells were treated with 10% FCS for 15, 36 and 48 hours, the group distribution at each stage of the cell cycle It is a figure which shows a mode. Shows time- and dose-dependent responses of Apioles to apoptosis of COLO 205 cells, and after synchronized COLO 205 cells were treated with 10% FCS for 15, 36 and 48 hours, the group distribution at each stage of the cell cycle It is a figure which shows a mode. Shows time- and dose-dependent responses of Apioles to apoptosis of COLO 205 cells, and after synchronized COLO 205 cells were treated with 10% FCS for 15, 36 and 48 hours, the group distribution at each stage of the cell cycle It is a figure which shows a mode. Shows time- and dose-dependent responses of Apioles to apoptosis of COLO 205 cells, and after synchronized COLO 205 cells were treated with 10% FCS for 15, 36 and 48 hours, the group distribution at each stage of the cell cycle It is a figure which shows a mode. FIG. 6 shows the effect of apiol treatment on caspase and bax / bcl-2 ratio of COLO 205 cells. It is a figure which shows the influence which the process by an apiol has on the apoptosis regulatory protein of COLO205 cell. It is a figure which shows the path | route which transmits the information which an apiol induces cell cycle stagnation in a human COLO205 cell.

  In order to obtain new active compounds that can suppress the proliferative action of colorectal cancer cells by screening and optimization, Applicants have identified a series of ten derivatives of 4,7-dimethoxy-1,3-benzodioxole for humans. The in vitro inhibitory activity against colorectal cancer cells (COLO 205) was evaluated. These derivatives are based on a lead compound (SY-1) isolated from cow beetle. By analyzing the relationship between the structure and activity of these 10 kinds of compounds, the length of the alkyl group at the 5-position is important, and apiol having an allyl group as the substituent exhibits the most excellent inhibitory activity. There was found. The present invention demonstrates that apiol, an analog of SY-1, can inhibit the proliferation of COLO 205 cells but does not affect the proliferation of normal human colon epidermal cells (FHC). did. G0 / G1 cell cycle arrest induced by apiol (75-225 μM) has a clear relationship with increased levels of p53, p21 and p27 and decreased levels of cyclosporin D1 I understood that. On the other hand, with respect to apoptosis of COLO 205 cells, caspase-3, -8, -9 and bax / bcl-2 ratios cleaved by apiol treatment (> 150 μM) are markedly increased, analysis of DNA fragmentation and flow From the analysis of the sub-G1 peak (Sub-G1 peak) in cytometry analysis, it can be seen that DNA fragmentation occurs. These results prove that SY-1 can suppress the growth of human colorectal cancer cells, and in particular, apiol has a remarkable effect.

  In the present invention, 5-substituted 4,7-dimethoxy-1,3-benzodioxole and its enantiomers, non-enantiomers, and pharmaceutically acceptable salts are any known extract from natural products such as cow turf. It can be obtained by techniques or synthesized by synthesis of 4,7-dimethoxy-5-methyl-1,3-benzodioxole based on known chemical means.

  The present invention is further illustrated by the following examples, which should be clarified that these examples are for illustrative purposes only and should not be construed as practical limitations on the present invention.

Experimental materials and experimental methods Compounds and Reagents 5-Substituted 4,7-dimethoxy-1,3-benzodioxole (5-Substituted 4,7-dimethoxy-1,3-benzodioxoles, ie, SY-1 derivatives, used in the Examples below. And compounds 1-9) shown in Table 1 are from Aurora Fine Chemicals Ltd. of Austria. Purchased from. Apiole was purchased from Uni-onward Company (Taiwan). The structures of these compounds were identified by NMR spectra.

2. Cell Culture The COLO 205 (p53-wild type) cell line was isolated from human colon adenocarcinoma (CCL-222, American Type Culture Collection, USA). This cell line was cultured in RPMI-1640 medium (Invitrogen, Carsbad, Calif.) Containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin (Invitrogen, Carlsbad, Calif.). FHC (CRL-1831, American Type Culture Collection, USA) is a normal human embryonic colonic mucosal long term epidermal cell culture (Siddiqui KM and Chopra DP, In Vitro, 1984; 20: 859-68). The culture was cultured in an incubator at 37 ° C. containing 5% wet CO ₂ .

3. Cell viability assay
The growth inhibitory effect of the compound on the cultured cells is determined by 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl-tetrazolium bromide assay [3- (4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazole bromide assay, MTT assay]. Each well was cultured at a density of 2 × 10 ⁴ cells (cells / well) in a 24-well culture plate, allowed to stand overnight, and then treated with different concentrations of compounds. After 24, 48 and 72 hours of incubation, 30 μL of 2 mg · mL ⁻¹ MTT solution [phosphate buffered saline, PBS, adjusted to pH 7.4] was placed in each well, and the media plate was further replaced Incubate for 3 hours. Upon incubation, it was gently absorbed and the medium was removed from each well and replaced with 200 μL dimethyl sulfoxide (DMSO). The absorption value of each well was measured at a wavelength of 550 nm by an enzyme-linked immunosorbent analyzer.

4). Flow cytometry analysis
After treating COLO 205 cells with different concentrations of apiol, they were absorbed with trypsin-ethylenediaminetetraacetic acid (trypsin-EDTA) after each different time, washed twice with PBS after filtration, 24% in 70% alcohol at 4 ° C. It was fixed over time. The fixed cells were filtered, washed again with PBS, treated with DNase-free RNase (2 U mL-1) for 30 minutes, and then propidium iodide reagent [3.8 mM sodium citrate (3.8 mM sodium citrate). cirate, 0.1% Triton X-100 and 20 mg ml-1 propidium iodide]. The DNA content was measured with a flow cytometry meter (Cytomics FC500, Beckman Coulter). The percentage of cells in each stage of the cell cycle was analyzed by software (eg, Multicycle 32 bit version, Beckman Coulter).

5. Immunoblotting analysis
The COLO 205 cell line was treated with different concentrations of apiol for different durations, followed by lysis buffer [20 mM Tris-HCl, 1% NP-40, 137 mM NaCl, 50 mM EDTA, protease inhibition. The resulting cell lysate was used for immunoblot analysis (Lin H et al., J Biol Chem, 2007; 282: 2776-84). ). Protein extracts obtained from treated cells (100 μg in each lane) are separated by SDS-PAGE, detected by specific antibodies, and protein loading is corrected by β-actin content. Immunodetection was performed overnight at 4 ° C using specific antibodies diluted appropriately. Primary antibody was diluted at a ratio of 1: 100 and 1: 1000. The primary antibodies include anti-cyclin D1 (anti-cyclin D1) and anti-p27 (Santa Cruz, Inc., CA, USA), anti-actin (Chemicon Co., MA, USA), anti-cleaved caspase- 3, -8, -9 (anti-cleaved caspase-3, -8, -9) (Cell Signaling Technology, Inc., MA, USA), anti-p21, anti-p53 (BD Bioscience, CA, USA) and anti-bax And anti-bcl-2 (Assay Designs, MI, USA). Secondary antibodies include goat anti-mouse or anti-rabbit antibody conjugated with peroxidase and goat anti-mouse antibody conjugated with alkaline phosphatase conjugate goat anti-mouse antibody (Jackson ImmunoResearch Laboratory, PA, USA), diluted 1: 5000 and incubated at room temperature for 1 hour. Trans-Blot SD (Bio-Rad Co., CA, USA) using ECL detection reagent and BCIP / NBT substrate solution (Perkin Elmer Co., MA, USA). ) Is used to visualize the immunoreactive proteins transferred on the PVDF membrane (PVDF membranes).

6). Analysis of DNA fragmentation (analysis of DNA fragmentation)
COLO 205 cells with different treatments were used to measure cell apoptosis by analysis of DNA fragmentation (Ho YS et al., Mol Carcinog, 1996; 16: 20-31). Genomic DNA was extracted and electrophoresed on a 2% agarose gel. DNA was visualized by ethidium bromide staining.

7). Statistics All numerical values are shown in the mean ± standard deviation (means ± SE) format. Significant comparisons were made by Dunnett's one-way ANOVA. Significance is bounded by P <0.05.

Cytotoxic activity of 5-substituted 4,7-dimethoxy-1,3-benzodioxole derivatives against human colon cancer cells
In this example, the anti-proliferation effect of 10 SY-1 derivatives was analyzed and COLO 205 cells were analyzed at different concentrations of each compound (37.5-225 μM) at different times (24, 48 and 72). Time), and then treated with 3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-tetrazolium bromide dimethoxy as described in “3. The assay was used to measure the cytotoxic activity of each compound.

The antiproliferative activity of each compound in COLO 205 cells is shown in FIGS. As a result, those in which the functional group at the 5-position is an aliphatic substituent (compounds 4, 5 and apiol) are relatively higher than those in which a polar alkane functional group is present (compounds 1 to 3). It was found to have an antiproliferative effect. The IC ₅₀ values of the compounds 2, 3, 4, 5 and 6 (IC ₅₀ value) as a result each 59.4Myu M measurement, 152.3μM, 148.5μM, 72.1μM, 110.9μM and 66. 7 μM. However, those with relatively bulky substituents at position 5 (compounds 7, 8 and 9) showed a very weak inhibitory effect (> 225 μM).

The results are as shown in FIGS. 1A-1C, but the most potential for COLO 205 cells is apiole, with IC ₅₀ values clearly <37.5 μM at 48 and 72 hours. (FIGS. 1B and 1C). The side chain of the SY-1 derivative is 2-propenyl group (Apiol) and n-propyl group (Compound 5), but apiol against COLO 205 cells The antiproliferative activity was more than three times that of those having a propyl group.

  In addition, in order to examine whether the inhibitory effect of SY-1 derivative on the growth of COLO 205 cancer cells is specific to cell type (normal human colonic epithelial cells). (FHC) was treated with different doses of apiol (as shown in FIG. 2) for different times, and the cytotoxic activity of each compound was measured by cell viability analysis described in “Experimental Materials and Experimental Methods”.

  As shown in FIG. 2, the cytotoxic activity of apiol has not been observed in normal human cell lines. Therefore, in the following examples, the present invention will be described with reference to apiol as an example.

Cell cycle arrest of human colon line carcinoma cells induced by SY-1 derivatives
In previous studies (Lien HM. Et al., 2009, same as above; Ho YS et al., Food Chem Toxicol, 2005; 43: 1483-95), COLO 205 cells were replaced with complete medium. After 15 hours (15 h), the group-specific G0 / G1 cell group treated with the compound shows the greatest difference relative to the control group. Therefore, in this example, this time (15 h) was selected to examine the dose-dependent effect of cell cycle stasis due to apiol. This example shows the percentage of cells in each stage of the cell cycle of COLO 205 cells treated with different doses of apiol by analysis of “4. Flow cytometry analysis” described in “Experimental materials and methods”. Yes. The above-mentioned “5. Immunoblot analysis” revealed the expression status of COLO 205 treated with p21, p27 and p53 and cyclin D1 in cells.

  As shown in FIG. 3, apiol has a dose-dependent effect on G0 / G1 stagnation, among which significant G0 / G1 stagnation (significant G0 / G1 arrest) is apiol (> 75 μM). Induced in COLO 205 cells treated with, the effect is dose dependent.

  In previous studies, cyclin-dependent kinase inhibitors (CDKi) such as p21 and p27 are up-regulated in human colon cancer cells stagnating in G0 / G1 by anti-cancer agents. (Lien HM. Et al., 2009, ibid .; Ho YS et al., 2005, ibid .; Wu CH et al., Toxicol Appl Pharmacol, 2002; 180: 22-35). Has been revealed. In this example, the effect of apiol on the expression of cell cycle regulatory proteins was analyzed. The results are shown in FIG. 4 and are exposed to the above apiol (<150 μM) for 15 hours. In elapsed COLO 205 cells, expression of p21, p27, p53 and cyclin D1 was induced.

Caspase activation of human colon line cancer cells by SY-1 derivative This example further analyzed the status of apoptosis after COLO 205 cells were exposed to apiol for 36-48 hours. It was carried out according to “3. Flow cytometry analysis”, “4. Immunoblot analysis” and “5.

  First, the time-dependent dose effect of apiol cell cycle regulation was measured by flow cytometric cytometric analysis with reference to the above examples. The results are as shown in FIGS. 5A-5D, with high doses of apiol (150, 225 μM) increasing the significant sub-G1 peak (sub-G1 peak) group at 36 and 48 hours.

  Next, in the prior art, activated caspases (Thornberry NA and Lazebnik Y, Science, 1998; 281: 1312-6; el-Deity WS et al., Cell, 1993; 75: 817-25) are used in the development of apoptosis. It is shown that it is necessary. Therefore, Applicants further determined by immunoblot analysis the status of caspase activation and the involvement in the bax / bcl-2 ratio in COLO 205 cells after treatment with apiol, with different doses of A protein extract of COLO 205 cells treated with apiol [100 μg in each lane] was separated by SDS-PAGE and detected with a specific antibody, and the protein load was calibrated with β-actin.

  As shown in FIG. 5E, apiol (150 or 225 μM, 36 hours) induces COLO 205 apoptosis, accompanied by an increase in the bax / bcl-2 ratio and caspase-3, -8, -9 cleavage. .

  Next, the applicant examined the apoptotic status of COLO 205 cells treated with apiol by analyzing the DNA fragmentation effect. DNA fragmentation was observed only in COLO 205 cells after 36 hours of treatment with apiol (> 75 μM). It can be seen that the DNA fragmentation effect is indeed induced by a high dose of apiol (150 μM), as shown in FIG.

  From the above, for example, SY-1 derivatives such as apiol certainly induces apoptosis of human colon line cancer cells specifically for the cell type and can therefore be used for colorectal cancer chemotherapy.

Claims (14)

A method for producing a pharmaceutical product for treating colorectal cancer using 5-substituted 4,7-dimethoxy-1,3-benzodioxole, comprising the following general formula (I):
(In the above formula, R represents a substituted or non-substituted, alkyl group having 1 to 6 carbon atoms, an alkenyl group having 2 to 6 carbon atoms, An alkynyl group having 2 to 6 carbon atoms, a cyano group, an ester group having 1 to 5 carbon atoms, a carboxylic group having 1 to 5 carbon atoms, an iso group A 5-substituted 4, which is a group selected from the group consisting of an oxazole group, provided that (with the proviso tat) R is not a methyl group (other than —CH 3) 7 Method for producing a colorectal cancer treatment drugs with dimethoxy-1,3-benzodioxole.   R is a substituted or unsubstituted ethyl group, propyl group, vinyl group, allyl group, butadienyl group, ethynyl group, propynyl group, cyano group, methyl ester group, ethyl ester group or carboxymethyl group, carboxyethyl group The method according to claim 1, which is a carboxylpropyl group.   The production method according to claim 1, wherein R is a substituted group, and the substituent is a hydroxy group (—OH), a cyano group (—CN), a nitro group (nitro group), or a halogen group (halogenyl). R is —CN, —COOH, —CH ₂ CH ₂ OH, —COOCH ₃ , — (CH ₂ ) ₂ CH ₃ , —CH ₂ CH (OH) CH ₂ CN, —CH ₂ CH═CH ₂ ,
and
The manufacturing method of Claim 1 selected from the group consisting of.   5-Substituted 4,7-dimethoxy-1,3-benzodioxole is intravenous, intramuscular, intraperitoneal, subcutaneous, rectal, topical, topical 5) The production method according to any one of claims 1 to 4, which is administered via a route.   The 5-substituted 4,7-dimethoxy-1,3-benzodioxole is dosed in an effective amount ranging from about 0.1 mg / kg to 500 mg / kg. The manufacturing method as described. A therapeutically effective amount of any of the aforementioned 5-substituted 4,7-dimethoxy-1,3-benzodioxole and enantiomers, non-enantiomers, pharmaceutically acceptable salts, or combinations thereof;
A pharmaceutical composition for use in the treatment of colorectal cancer, comprising these pharmaceutically acceptable carriers and / or excipients,
The 5-substituted 4,7-dimethoxy-1,3-benzodioxole is represented by the following general formula (I)
(In the above formula, R represents a substituted or non-substituted, alkyl group having 1 to 6 carbon atoms, an alkenyl group having 2 to 6 carbon atoms, An alkynyl group having 2 to 6 carbon atoms, a cyano group, an ester group having 1 to 5 carbon atoms, a carboxylic group having 1 to 5 carbon atoms, an iso group A pharmaceutical composition represented by a group selected from the group consisting of an oxazole group (provided that (with the proviso tat) R is not a methyl group (other than -CH3))   R is a substituted or unsubstituted ethyl group, propyl group, vinyl group, allyl group, butadienyl group, ethynyl group, propynyl group, cyano group, methyl ester group, ethyl ester group, carboxymethyl group, carboxyethyl group The pharmaceutical composition according to claim 7, which is a carboxylpropyl group. R is —CN, —COOH, —CH ₂ CH ₂ OH, —COOCH ₃ , — (CH ₂ ) ₂ CH ₃ , —CH ₂ CH (OH) CH ₂ CN, —CH ₂ CH═CH ₂ ,
and
The pharmaceutical composition according to claim 7, which is selected from the group consisting of: The pharmaceutical composition according to claim 7, which is administered orally or parenterally.   8. The pharmaceutical composition according to claim 7, wherein the carrier is a low melting point wax.   11. The effective amount range of 5-substituted 4,7-dimethoxy-1,3-benzodioxole is from about 0.1 mg / Kg to about 500 mg / Kg, according to any one of claims 7-10. Pharmaceutical composition.   A cell cycle inhibitor comprising the 5-substituted 4,7-dimethoxy-1,3-benzodioxole according to any one of claims 1 to 4.   An agent for killing colorectal cancer cells, comprising the 5-substituted 4,7-dimethoxy-1,3-benzodioxole according to any one of claims 1 to 4.