CN103483187B - 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid and Pharmaceutical composition thereof and application - Google Patents

4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid and Pharmaceutical composition thereof and application Download PDF

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CN103483187B
CN103483187B CN201310390817.XA CN201310390817A CN103483187B CN 103483187 B CN103483187 B CN 103483187B CN 201310390817 A CN201310390817 A CN 201310390817A CN 103483187 B CN103483187 B CN 103483187B
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cancer
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贺小琼
詹华强
黄清龙
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Hong Kong University of Science and Technology HKUST
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Abstract

The invention discloses and there is 4 ethyoxyl 2 hydroxyl 6 methyl benzoic acids of formula (1) structure and Pharmaceutical composition thereof and application.Kill with external equal energy conspicuousness and/or suppress the kinds of tumor cells of human and animal in this compound body, this compound and clinical antitumor agents have conspicuousness synergistic antitumor effect, and this compound or its pharmaceutically acceptable salt can be applicable to prepare anti-tumor drug, antineoplastic pharmaceutical compositions, combined with antineoplastic compositions or adjunct antineoplastic health products.This compound energy conspicuousness promotes cancer cell-apoptosis and necrosis, regulation growth of cancer cells cycle, inhibition cancer cell propagation.The body weight of mouse, under effective antitumor dosage, is increased and internal organs is grown without conspicuousness toxicity, or toxic action conspicuousness is less than clinical anti-cancer medicine such as endoxan and 5 fluorouracils by this compound.

Description

4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid and Pharmaceutical composition thereof and application
Technical field
The invention belongs to chemical medicine, particularly relate to a kind of 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid and Pharmaceutical composition and application.
Background technology
Cancer is a major disease of serious threat human health and life security, has the most become first of the mankind The cause of the death.According to whole world cancer statistical report, global new cancer cases in 2008 is 1266.1 ten thousand examples, dead 756.4 ten thousand examples；China Cancer new cases are 281.6 ten thousand examples, dead 195.8 ten thousand examples." 2012 China's tumour registration annual report " discloses, and Chinese 2012 new Sending out cases of cancer 3,120,000, average minute clock has 6 people to be diagnosed as malignant tumour.The incidence of disease of cancer is also rising, and cancer becomes Serious threat people's health and a Major health problem of life security.
Cancer is still a kind of obstinate disease the most at present.The chemotherapy of cancer is arranged as systemic treatment Execute, the treatment of cancer occupies epochmaking status, be also the hope the most thoroughly solving treatment of cancer problem.
The most currently used cancer chemotherapeutic drug, a lot of medicines exist that toxic and side effect is big, uncertain therapeutic efficacy is fixed, selectivity Difference and the problem such as cancer cell resistance, make the application clinically of cancer chemotherapy drug therapy and effect be very limited.Cause This, exploitation clear curative effect, safety low-poison, particularly there is the chemotherapeutics of selective antitumaous effect, be that PTS is studied One important directions and urgent task.
Summary of the invention
Based on this, it is an object of the invention to provide a kind of 4-ethyoxyl-2-hydroxyl-6-methylbenzene with antitumor action Formic acid.
Concrete technical scheme is as follows:
There is the 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid (4-ethoxy-2-hydroxy-6-of formula (1) structure Methylbenzoic acid) or pharmaceutically acceptable salt,
It is a further object of the present invention to provide a kind of antitumor medicine composition.
Concrete technical scheme is as follows:
A kind of antitumor medicine composition, its active component includes above-mentioned 4-ethyoxyl-2-hydroxyl-6-methylbenzene first Acid or its pharmaceutically acceptable salt.
Wherein in some embodiments, the formulation of this pharmaceutical composition is: parenteral solution, powder-injection, tablet, oral liquid, glue Capsule, granule or electuary.
It is a further object of the present invention to provide a kind of associating antitumor medicine composition.
Concrete technical scheme is as follows:
A kind of associating antitumor medicine composition, its active component includes above-mentioned 4-ethyoxyl-2-hydroxyl-6-methylbenzene At least one in formic acid or its pharmaceutically acceptable salt, and 5 FU 5 fluorouracil, taxol, cis-platinum or endoxan.
It is a further object of the present invention to provide the application of above-mentioned 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid.
Concrete technical scheme is as follows:
Above-mentioned 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid or its pharmaceutically acceptable salt are preparing antineoplastic In application.
Wherein in some embodiments, described tumour is any one in following human tumor: human breast carcinoma, prostate Cancer, lung cancer, cancer of the stomach, cervical carcinoma, colon cancer, liver cancer, oophoroma, leukaemia, neuroma, sarcoma.
It is a further object of the present invention to provide above-mentioned 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid to protect at adjunct antineoplastic Application in strong product.
Concrete technical scheme is as follows:
Above-mentioned 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid or its pharmaceutically acceptable salt keep healthy at adjunct antineoplastic Application in product.
A kind of adjunct antineoplastic health products, its active component include above-mentioned 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid or Its pharmaceutically acceptable salt.The formulation of these adjunct antineoplastic health products is: tablet, oral liquid, capsule, granule or electuary.
Advantages of the present invention is as follows:
4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid (4-ethoxy-2-hydroxy-6-methylbenzoic of the present invention Acid) compound is new construction material, can carry out chemical synthesis by phase-transfer-catalyzed reactions；This compound or its salt class can As antitumor chemotherapeutic agent, or the antineoplastic pharmaceutical compositions as antitumor activity composition, and as auxiliary Help anti-tumor health care product.4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid is external conspicuousness can be killed and/or suppress the mankind and moves The multiple cancer cell of thing, such as human breast carcinoma, prostate cancer, lung cancer, cancer of the stomach, cervical carcinoma, colon cancer, HCC, and mouse Sarcoma, liver cancer, lung carcinoma cell etc..To balb/c nude mice model Human Lung Cancer and human liver cancer in this compound body, and Kunming Murine sarcoma that kind of mouse and ICR mouse are transplanted and rat liver cancer, the tumour such as the mice lung cancer that C57BL/6J mouse is transplanted, all have There is the growth inhibition effect of conspicuousness.This compound and the antineoplastic chemotherapy medicine of Clinical practice in vitro and/or internal all have There is combination and cooperation antitumor action.This compound has selective antitumaous effect, and the sensitiveness of different carcinoma cell line is different, liver cancer Cell is very sensitive to this compound.This compound energy conspicuousness promotes cancer cell-apoptosis, regulation Cancer Cell cycle and differentiation, presses down The propagation of cancer cell processed.The body weight of mouse, under effective antitumor dosage, is increased and internal organs is grown without conspicuousness poison by this compound Property, or toxic action conspicuousness is less than clinical anti-cancer medicine such as cyclophosphamide and 5-fluorouracil.
Accompanying drawing explanation
Fig. 1 is the mass spectrogram of formula (1) compound；
Fig. 2 is the nucleus magnetic hydrogen spectrum figure of formula (1) compound；
Fig. 3 is nuclear-magnetism carbon spectrogram and the DEPT spectrogram of formula (1) compound；
Fig. 4 is formula (1) compound dose-effect curve figure to different tumor cell lines；
Fig. 5 is time m-effect, the dose-effect relationship curve map of formula (1) Compound ira vitro anti-tumor experiment；
When Fig. 6 is antitumor (NCI-H460) test in formula (1) compound body, the tumour of different time points is relative Volume (RTV)；
When Fig. 7 is antitumor (NCI-H460) test in formula (1) compound body, the tumour of different time points is relative Appreciation rate (%)；
Fig. 8 is the NCI-H460 tumor weight that formula (1) compound is administered after 10 times；
When Fig. 9 is antitumor (QGY-7703) test in formula (1) compound body, the tumour of different time points is relative Volume (RTV)；
When Figure 10 is antitumor (QGY-7703) test in formula (1) compound body, the tumour phase of different time points To the rate of increase (%)；
Figure 11 is in formula (1) compound body at the end of antitumor (QGY-7703) experiment, and the tumour of each group is average Weight；
Figure 12 is in formula (1) compound body at the end of antitumor (QGY-7703) experiment, the tumour of each treated animal Photo；
Figure 13 is in formula (1) compound body at the end of antitumor (QGY-7703) experiment, the spleen of each treated animal Photo；
Figure 14 is the flow cytomery result (QGY-7703) of formula (1) compound solvent comparison；
Figure 15 is the flow cytomery result (QGY-7703) of formula (1) compound；
Figure 16 is formula (1) compound (D4,50 μMs) the Western Blot testing result at QGY-7703；
Figure 17 is formula (1) compound (D1,200 μMs) the Western Blot testing result at HCT-116；
Figure 18 is formula (1) compound (D2,150 μM) the Western Blot testing result at MCF-7；
Figure 19 is formula (1) compound (D1,200 μM) the Western Blot testing result at A-549；
Figure 20 is that formula (1) compound combines anti-A-549 function of tumor with the external of 5 FU 5 fluorouracil；
Figure 21 is that formula (1) compound combines anti-A-549 function of tumor with the external of cis-platinum.
Detailed description of the invention
The present invention is further elaborated by the following examples.
The embodiment 1 :-2-hydroxyl-6-methyl benzoic acid (4-ethoxy-2-of compound 4-ethyoxyl shown in formula (1) Hydroxy-6-methylbenzoic acid) synthesis and Structural Identification
1, preparation method
Phase-transfer Wittig reaction and column chromatography for separation
Utilize phase-transfer-catalyzed reactions, in the presence of NaOH, 2,4-dihydroxy-6-methyl benzoic acids and sulfuric acid diethyl Ester (mol ratio is 0.2:0.18), at the double octadecyl methyl amine benzyl chloride quaternary ammonium salt of catalyst HA-1() under effect in 60-65 DEG C reaction 3h.After completion of the reaction, it is cooled to 25 DEG C, pours in frozen water, adjust pH value to neutral with watery hydrochloric acid, separate organic layer；Water Layer toluene extracts 2 times, merges organic layer；Washing organic layer 3 times, anhydrous sodium sulfate is dried.Decompression boils off solvent toluene, product Use silica gel column chromatography technology to carry out again isolated and purified, obtain compound 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid.
2, Structural Identification
The mass spectrogram (as shown in Figure 1) of compound 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid；
The nucleus magnetic hydrogen spectrum figure (as shown in Figure 2) of compound 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid；
The nuclear-magnetism carbon spectrogram of compound 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid and DEPT spectrogram (as shown in Figure 3).
Embodiment 2: the biological experiment result of formula (1) compound 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid
Formula (1) the compound 4-ethyoxyl-2-hydroxyl-6-first of the present invention is described below by part specific experiment illustration Yl benzoic acid (following chart represents with D6B2 or D, lower same；D1 is 200 μMs, D2 is 150 μMs, D3 is 100 μMs, D4 is 50 μMs, D5 is 25 μMs) there is conspicuousness and selective antitumor action and the synergistic antitumor effect of clinical antitumor agents, Yi Jiqi Toxicological safety, Antitumor Mechanism.
1, the anti-cancer effect in vitro of 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid:
Test method: formula (1) compound 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid dissolves with DMSO, by ultimate density It is 25,50,100,150,200 μMs and carries out external anticancer test.Collect the exponential phase mankind or animal tumor cell, with completely After culture medium suspendible, being inoculated in 6 orifice plates, every hole adds 2ml cell suspension.37℃、5%CO2After incubator is cultivated 24 hours, take 3 Porocyte counts the cell number as 0 hour；Suck the nutrient solution in remaining hole, be then separately added into containing respective concentration by design The complete culture solution 2ml of formula (1) compound, sets DMSO solvent control simultaneously.It is parallel, carefully that every concentration and DMSO control group do 9 holes Born of the same parents after dosing in 37 DEG C, 5%CO2Incubator continues to cultivate.24h, 48h, 72h after dosing, it is thin that every concentration collects 3 holes respectively Born of the same parents count, and calculate mean value and the standard deviation of cell number, draw cell growth curve, carry out significant difference inspection.With Shi Caiyong 96 orifice plate is cultivated, and measures the OD value after drug-treated 24,48 and 72 hours, the time m-effect of observation medicine with mtt assay Relation.
Result: formula (1) compound 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid is to MCF-7 cell strainHJ2mm, people It is anticancer that lung cancer cell line A-549, human colon cancer cell strain HCT-116, human hepatoma cell strain QGY-7703 etc. are respectively provided with conspicuousness Effect.This compound has significant difference to the inhibiting rate of different carcinoma cell, and human hepatocarcinoma cells QGY-7703 is to this compound Very sensitive.200 μMs of concentration, 72 hours, the inhibiting rate of QGY-7703, MCF-7, A-549 and HCT-116 is divided by this compound It is not 95.07%, 78.26%, 50.39% and 51.84%, there is obvious dose-response relationship, its half-inhibition concentration (IC50) It is respectively 70 μMs, 135 μMs, 190 μMs and 200 μMs.The external anti-cancer result of 72 hours is shown in Fig. 4.Sieve at other Anticancer Activity in vitro In choosing test, formula (1) compound treatment cancer cell, after 72 hours, is shown under the microscope to human cervical carcinoma cell Hela, people's cancer of the stomach Cell SGC-7901, Human Prostate Cancer Cells PC-3, Proliferation of Human Ovarian Cell SKOV-3, human leukemia cell K-562 and HL-60, Human glioma cell U-251 etc. has the external suppression of conspicuousness or these cell lines (are not entered by killing action at present temporarily Row dose-effect relationship and time m-effect relation research).
This compound dose-effect relationship figure to human liver cancer cell QGY-7703 anti-cancer effect in vitro at different dosages With time m-effect relation figure (mtt assay) see Fig. 5.
2,4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid mankind's maxicell lung cancer NCI-to balb/c nude mice model The internal antitumaous effect of H460
Test method: male balb/c nude mice (body weight 16～18g), every right side dorsal sc inoculation mankind's maxicell lung Cancer NCI-H460 cell suspension 0.1ml(5 × 107Cells/ml).Inoculation cancer cell after the 12nd day, major part mouse subcutaneous go out Existing naked eyes visual tumors.With vernier caliper measurement, the gross tumor volume that calculates every mouse.Tumor-bearing mice is pressed gross tumor volume size It is randomly divided into DMSO control group and formula (1) compound administration group, often 8 tumor-bearing mices of group.Formula (1) compound is with aseptic mass spectrum Pure DMSO prepares, and dosage is 30mg/kg；Control group mice gives the DMSO of equal volume.Mouse from packet after second It starts intraperitoneal injection, is administered once every other day, within every 4 days, measures 1 gross tumor volume and body weight, calculate tumour relative volume, The relative appreciation rate of tumour.After being administered 10 times, put to death mouse, completely strip tumour, liver,spleen,kidney and testis, measure tumor weight And each organ weights, calculate tumor weight inhibiting rate and each organ coefficient.
Result: the suppression result of NCI-H460 tumour is shown in Fig. 6, Fig. 7, figure by 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid 8.The growth (p < 0.05) of 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid energy conspicuousness suppression Human Lung Cancer NCI-H460, test At the end of, the weight inhibiting rate of tumour is 43.51%, and the relative appreciation rate of tumour is 51.96%, the tumor weight of administration group, swollen Knurl relative volume compares with negative control group (representing with NC, lower same), declines (p < 0.05) in conspicuousness.
3,4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid is to transplantability human liver cancer QGY-7703 anticancer in nude mouse Effect
Test method: to raising the female balb/c nude mice (body weight 16～18g) of 1 week through adaptability, carries on the back on right side for every Portion's subcutaneous vaccination QGY-7703 cell suspension 0.1ml(1 × 108Cells/ml).Nude mice is raised in SPF laboratory.Inoculation cancer is thin After born of the same parents the 21st day, mouse is subcutaneous there is naked eyes visual tumors.With major diameter and the transverse diameter of vernier caliper measurement mouse tumor, calculate Gross tumor volume.Tumor-bearing mice is randomly divided into DMSO solvent control group, 5 FU 5 fluorouracil (5-Fu) sun according to gross tumor volume size Property drug control group and formula (1) compound administration group, often organize totally 6 tumor-bearing mices.4-ethyoxyl-2-hydroxyl-6-methylbenzene first After acid is dissolved with the aseptic pure DMSO of mass spectrum, then diluting with the aseptic PBS of equal-volume, dosage is 20mg/kg；Solvent control group Mouse gives the DMSO/PBS(1:1 of equal volume)；5-Fu dilutes with equal-volume DMSO, and 5-Fu dosage is 30mg/kg.Little The mouse second day beginning intraperitoneal injection after packet, is administered once every other day, within every 4 days, measures 1 gross tumor volume and body weight.Give After medicine 12 times, put to death mouse, completely strip tumour, liver,spleen,kidney, measure tumor weight, Mouse Weight and each organ weights.
Result: formula (1) the compound 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid suppression result to QGY-7703 tumour See Fig. 9, Figure 10, Figure 11.The growth (p < 0.05) of formula (1) compound energy conspicuousness suppression human liver cancer QGY-7703, test knot Shu Shi, the weight inhibiting rate of tumour is 69.83%, and the relative appreciation rate of tumour is 30.84%, the tumor weight of administration group, tumour Relative volume compares with control group, declines (p < 0.05) in conspicuousness.This compound inhibitory action conspicuousness to human liver cancer It is better than the inhibitory action to lung cancer.At the end of experiment, Figure 12 is shown in by the QGY-7703 tumour photo of each group.
Murine sarcoma S-180 that Kunming mouse and ICR mouse are transplanted by 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid, Rat liver cancer H-22, and C57BL/6J mouse transplant mice lung cancer 3ll be respectively provided with conspicuousness internal antitumaous effect (p < , and there is dose-effect relationship 0.05).
4, the 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid toxic action to nude mice
Experimental technique: in nude mouse in anticancer test, is measuring each time point of gross tumor volume, is measuring the body of mouse simultaneously Weight；When off-test, put to death mouse, completely take out mouse liver, spleen, kidney, testis (male mouse) and weigh, calculating each dirty Device coefficient.
(1) impact that male mouse body weight and internal organs are grown
The body weight (g) of table 1. male balb/c mouse different time points
*: P < 0.05
Result: after being administered 4 times, Mouse Weight is increased by formula (1) compound 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid Present inhibitory action.
The organ coefficient of table 2. male balb/c mouse
*:P<0.05
Result: formula (1) compound 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid can affect liver and the spleen of male nude mouse Dirty growth, but do not affect kidney and the testicualr development of male mice.
(2) impact that, body weight dead on female mice and internal organs are grown
The body weight (g) of table 3. female balb/c mouse different time points
2 mouse of #:5-fluorouracil group are dead at the 20th day, separately have 3 extremes to become thin, dying；
*: from the beginning of the 8th day, 5 FU 5 fluorouracil and D6B2 group body weight conspicuousness are less than solvent control group, P < 0.05；
Result: cancer therapy drug 5 FU 5 fluorouracil and formula (1) compound energy conspicuousness affect Mouse Weight and grow, 5-fluorine urine is phonetic Pyridine affects conspicuousness higher than this compound to Mouse Weight.At the 20th day of experiment, 5-Fu group had 2 animals dead because of exhaustion, Separately there are 3 mouse extreme exhaustions and dying.Formula (1) compound treated animal is still bouncing, nothing death when off-test.
The organ coefficient of table 4. female balb/c mouse
*: P < 0.05
Result: 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid can affect female mice kidney development, and 5 FU 5 fluorouracil Energy conspicuousness affects spleen and the kidney development (Figure 13 is shown in by the spleen photo of each group mouse) of mouse.This compound is to Mouse Weight It is less than cancer therapy drug 5 FU 5 fluorouracil with the development impact conspicuousness of internal organs.
5,4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid is on apoptosis of tumor cells and the impact of cell cycle:
Experimental technique: HCC QGY-7703, after formula (1) compound treatment 48h, uses flow cytometer (FACS) Detection cell cycle and Apoptosis situation.Result is shown in Figure 14 and Figure 15.
Result: formula (1) compound energy conspicuousness promotes apoptosis and the necrosis of cancer cell, and drug-treated group is 100 μ at dosage When M, 150 μMs and 200 μMs, the DNA fragment peak (P2) that Apoptosis and necrosis is formed is respectively 13.5%, 24.7% and 32.7%, has Significantly dose-effect relationship, conspicuousness is higher than the 6.8% of control group, and this compound conspicuousness postpones the G0/ of cancer cell G1 and G2/M peak value and cell cycle distribution ratio, hence it is evident that affect differentiation and the cycle of cancer cell.
6,4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid to Apoptosis marker protein Cleaved Caspase-3 and The impact that Cleaved PARP level and PTEN Tumor Suppressor Gene are expressed:
Experimental technique: HCC QGY-7703, colon cancer cell HCT-116, breast cancer cell MCF-7, lung carcinoma cell A-549, after formula (1) compound treatment 24 hours, collects cell, cracks, adjusts protein concentration, use Western Blot skill Art measures.Result is shown in that (SP represents positive control Staurosporine, and SP1 is 50nM, SP2 for Figure 16, Figure 17, Figure 18, Figure 19 For 100nM).
Result: 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid energy conspicuousness promotes HCC QGY-7703, colon cancer The formation of Apoptosis mark primer Cleaved PARP in cell HCT-116, to breast cancer cell MCF-7, lung carcinoma cell A- In 549, the formation of Cleaved PARP is without effect.This compound nor affect on cancer cell Cleaved Caspase-3 level and The expression of PTEN.
7,4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid and the synergistic antitumor effect of other cancer therapy drug:
In vitro in anticancer test, compound 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid is phonetic with cancer therapy drug 5-fluorine urine Pyridine, taxol and cis-platinum have the external synergistic antitumor effect of conspicuousness.The results are shown in Table 5, table 6 and Figure 20, Figure 21.
Test method: in external anticancer test, arranges negative control group, the positive cancer therapy drug group of a dosage, 3 agent The D6B2 group of amount and the D6B2 of three dosage add positive cancer therapy drug group, within 72 hours after dosing, collect cell count, and calculating presses down Rate processed and carry out statistical analysis.
Table 5.D6B2 combines the 5-Fu In-vitro Inhibitory Effect (n=3, x ± s) to people's lung cancer cell A-549
Compare with control group,*P ＜ 0.05,**P ＜ 0.01；Compare with 5-FU,△P ＜ 0.05,△△P ＜ 0.01.
Table 6D6B2 combination with cisplatin (DDP) to the inhibitory action of lung cancer cell A-549 (n=3,)
Compare with control group,*P ＜ 0.05,**P ＜ 0.01；Compare with DDP,△P ＜ 0.05,△△P ＜ 0.01.
Taxol is when 1nM concentration, and the external inhibiting rate to MCF-7 is 28.85%, and D6B2 is when concentration is 100 μMs pair The inhibiting rate of MCF-7 is 15.38%, and during the two synergy, the inhibiting rate to MCF-7 is 52.16%, 4-ethyoxyl-2-hydroxyl- 6-methyl benzoic acid and taxol have conspicuousness combination and cooperation anti-cancer effect in vitro in vitro to human breast cancer cell line Bcap-37.
In vivo in anticancer test, 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid has in vivo with endoxan combines Suppression rat liver cancer H-22.Endoxan is when 40mg/kg, and the inhibiting rate to rat liver cancer H-22 is 75.83%, and D6B2 exists During 40mg/kg, the inhibiting rate to H-22 is 38.99%, when the two is used in combination, combines group mouse and there are no tumor growth.
Embodiment 3:
Formula (1) compound 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid is neutralized in a heated condition with sodium acid carbonate Reaction, then adjusts pH value to 7.0-7.5 with soda ash solution, heats, evaporates, crystallizes the sodium salt of the formula that i.e. formed (1) compound Sodium4-ethoxy-2-hydroxy-6-methylbenzoate, can be dissolved in water.Formula (1) compound also can be with other alkalescence Ion or group form salt.
Embodiment 4:
Formula (1) compound 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid, dissolves with a small amount of DMSO, injects use routinely Water, essence filter, parenteral solution is made in embedding sterilizing.
Embodiment 5:
The sodium salt Sodium4-ethoxy-2-hydroxy-6-methylbenzoate of formula (1) compound, fills routinely Penetrating and dissolve with water, essence filter, parenteral solution is made in embedding sterilizing.
Embodiment 6:
Formula (1) compound 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid or its esters, with a small amount of ethanol, methyl alcohol or distillation After water dissolves, filter with aseptic suction funnel, more aseptic essence filter, it being sub-packed in ampoule, after frozen drying, aseptic sealing by fusing obtains Powder-injection.
Embodiment 7:
Formula (1) compound 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid or its esters, the ratio needed in preparation adds Excipient, makes pulvis.
Embodiment 8:
Formula (1) compound 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid or its esters, the ratio needed in preparation adds Excipient, pelletizing press sheet makes tablet.
Embodiment 9:
Formula (1) compound 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid or its esters, oral liquid preparation method is made routinely Oral liquid.
Embodiment 10:
Formula (1) compound 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid or its esters, the ratio needed in preparation adds Excipient, makes capsule or granule or electuary.
Embodiment 11:
Formula (1) compound 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid or its esters, added in the ratio of product needed Food or other carrier, make health products or other functional articles.
Embodiment 12:
Formula (1) compound 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid or its esters, needed by treatment, anti-swollen with other Tumor medicine mixes in proportion, makes various preparation, forms combined with antineoplastic compositions.
Embodiment described above only part have expressed the several embodiments of the present invention, and it describes more concrete and detailed, But therefore can not be interpreted as the restriction to the scope of the claims of the present invention.It should be pointed out that, for the ordinary skill of this area For personnel, without departing from the inventive concept of the premise, it is also possible to make some deformation and improvement, these broadly fall into the present invention Protection domain.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.

Claims (4)

1. an associating antitumor medicine composition, it is characterised in that its active component includes the 4-second with formula (1) structure Epoxide-2-hydroxyl-6-methyl benzoic acid or its pharmaceutically acceptable salt, and 5 FU 5 fluorouracil, taxol, cis-platinum or ring phosphinylidyne At least one in amine,
2. 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid or its pharmaceutically acceptable salt with formula (1) structure are anti-in preparation Application in tumour medicine,
Application the most according to claim 2, it is characterised in that described tumour is any one in following human tumor: people Breast cancer, prostate cancer, lung cancer, cancer of the stomach, cervical carcinoma, colon cancer, liver cancer, oophoroma, leukaemia, neuroma, sarcoma.
4. have the 4-ethyoxyl-2-hydroxyl-6-methyl benzoic acid of formula (1) structure or its pharmaceutically acceptable salt prepare auxiliary Help the application in anti-tumor health care product,
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