JPH0565507B2 - - Google Patents

Description

[Detailed description of the invention]

[Industrial Application Field] The present invention relates to an indole derivative and an anticancer drug effect enhancer containing the same as an active ingredient. [Prior Art and Problems] The number of cancer patients is increasing year by year, and the mortality rate due to cancer ranks first in Japan, and there is a high social interest in cancer treatment. Research and development of anticancer drugs for the treatment of cancer has been actively carried out, and various anticancer drugs are used clinically for the treatment of cancer. Although its effectiveness is steadily improving year by year, in many cases it is not always possible to achieve a satisfactory effect in completely suppressing cancer growth and maintaining the survival of cancer patients over a long period of time. Furthermore, many attempts are currently being made clinically to enhance the effects of anticancer drugs by combining multiple anticancer drugs (multidrug combination therapy). However, this case is also unsatisfactory as a chemotherapy method for cancer, and there is a strong need for the development of new cancer therapeutics from a new perspective. Under these circumstances, one possible method would be to develop more cooperative anticancer drugs and to develop a method for more selectively transporting anticancer drugs to target organs. These studies are currently being carried out all over the world, but the current situation is that they are becoming increasingly difficult. On the other hand, another important method is to try to enhance the effects of existing anticancer drugs, but it is particularly difficult to develop agents that enhance the effects of existing anticancer drugs against drug-resistant cancer, which is a serious clinical problem in cancer chemotherapy. considered as an important new cancer treatment method. The background of clinical resistance to anticancer drugs is not necessarily simple. Resistance in clinical practice can be broadly divided into two aspects. The first is when the cause is sought in an individual cancer patient, and the second is when the cause is sought in the cancer cells themselves. In recent years, the mechanism of resistance in this second case is being elucidated at the molecular level, and treatment methods for this are also being investigated. That is, a gene responsible for multidrug resistance has recently been isolated, and this gene is a membrane protein expressed in multidrug-resistant cells, P-glycoprotein (P-glycoprotein).
It was revealed that it was a gene for glycoprotein. P-glycoprotein is presumed to be a protein that has the function of excreting anticancer drugs from cells, and is considered to be a protein that plays a central role in the multidrug resistance mechanism. In addition, some mechanisms have been suggested to be common to cancers that are inherently difficult to respond to anticancer drugs, such as solid cancers. In other words, many anticancer drugs pass through the cell membrane and exert their effects within the cell, but in resistant cancer cells, the inflowing anticancer drug is excreted outside the cell by the action of this P-glycoprotein, and the drug inside the cancer cell is released. Concentrations are kept low. As a result, it is thought that the effects of anticancer drugs are less likely to be expressed. Therefore, the present inventors believe that, for example, a substance that suppresses the action of P-glycoprotein and inhibits the outflow of anticancer drugs from cancer cells has the effect of enhancing the effect of anticancer drugs, is particularly effective in overcoming resistance, and is a novel drug. We believe that it can be used as a cancer chemotherapeutic agent. In fact, Tsuruo et al. have shown that calcium antagonists such as verapamil block the outflow of anticancer drugs from cancer cells, and are therefore effective against resistant cancers in vitro and in vitro.
It has been found that it has the effect of enhancing the effects of anticancer drugs such as adriamycin and vincristine in vivo. However, when these calcium antagonists are used clinically in cancer patients, side effects such as a decrease in blood pressure and induction of arrhythmia appear, which poses a major problem as a cancer therapeutic agent. Therefore, there has been a desire for a drug that has a stronger effect of enhancing anticancer drug effects against resistant cancers and has fewer side effects. [Means for Solving the Problems] Based on the above viewpoint, the present inventors have made extensive studies and found that the indole derivatives shown below exhibit a strong anticancer drug effect enhancement effect in resistant cancers, and have low toxicity and low toxicity. They discovered that it has side effects and completed the present invention. That is, the present invention provides the general formula () (In the formula, R ₁ represents a hydrogen atom, a lower alkyl group, or an acyl group, and R ₂ represents a benzyl group, a diphenylmethyl group, a fluorenyl group, or a dibenzosuberanyl group. n represents 2 or 3.) The represented compounds and their salts (hereinafter referred to as compounds of the present invention) and anticancer drug effect enhancers contained as their active ingredients. Acyl group refers to acetyl group, propionyl group, benzoyl group, etc. Here, dibenzosuberanyl group refers to 10,11-dihydro-5H-dibenzo[a,b]cycloheptenyl group, also known as dibenzo [a, d] means a cyclohepta-1,4-dienyl group. Salts of the compounds of the present invention include salts with inorganic acids such as hydrochloric acid and sulfuric acid, or salts with organic acids such as acetic acid, oxalic acid, maleic acid, and tartaric acid. In addition, since the compound of the present invention has an asymmetric carbon in its structure, optical isomers exist, but the compound of the present invention shall contain all of these. Compounds structurally similar to indole derivatives include JP-A-56-32455, 57-146754,
There is a group of compounds described in 61-37765 etc. However, in these documents, there are no examples of the compound of the present invention itself. Furthermore, regarding the compounds shown in claim 2, specific physical property values, etc. are provided in Examples, etc.
There is no description that defines this compound at all, and there is no description of this compound in any of the documents other than these. Further, the above-mentioned document describes the use of cardiovascular drugs, particularly cardiotonic drugs, and does not provide any description regarding the anticancer drug effect enhancer mentioned in the present invention. From the above, all of the compounds of the present invention are new compounds, and we have discovered a completely new beneficial effect-enhancing effect of anticancer drugs, and we claim the rights of the present invention to these compounds. Next, as for the synthesis method of the compound of the present invention, the first method is to react an indole derivative represented by the following formula with a halide such as epichlorohydrin or epibromohydrin in the presence of a base, (In the formula, R ₁ is the same as above, and X represents a halogen.) This leads to a corresponding epoxy compound. Bases herein include inorganic bases such as sodium hydroxide, sodium hydride, potassium carbonate, potassium t-butoxy, and sodium carbonate, triethylamine,
Represents an organic base such as pyridine. As a solvent,
Water solvent or alcohol, acetone, THF,
Organic solvents such as DMF can be used, and the reaction temperature is 0~
A range of 100°C is preferred. Furthermore, the compound of the general formula () of the present invention can be obtained by thermally reacting a corresponding epoxy compound with a corresponding amine derivative. (In the formula, R ₁ , R ₂ , and n have the same meanings as above.) Thermal here means from room temperature to the boiling point of the solvent used, and examples of the solvent include alcohol, acetone, chloroform, Organic solvents such as DMF are used. The effect of enhancing anticancer drug effect on resistant cancers of the compound of the present invention was determined by using adriamycin-resistant human ovarian cancer cell line 2780AD or adriamycin-resistant human myeloid leukemia cell line K562/ADM, and the effect of enhancing the anticancer drug retention in the cells and the anticancer drug. This is evidenced by the action-enhancing effect of As detailed in the test examples, the compounds of the present invention all showed remarkable anticancer drug retention enhancing effects and anticancer drug action enhancing effects. Among them, the compounds shown in Examples 1 and 4 showed particularly strong effects. Further, the anticancer agent to be used in combination with the compound of the present invention or its salt is not particularly limited, but preferable ones include non-antimetabolite anthracycline antibiotics, such as adriamycin, daunomycin, aclacinomycin A; Mycin antibiotics, such as actinomycin C, actinomycin D; chromomycin antibiotics,
For example, mithramycin, toyomycin; vinca alkaloids, such as vincustine, vinblastine; maytansines; vodophyllotoxin derivatives, such as VP16-213; homohalintonin; anguidein; bruceantin; neocarzinostatin; anthramycin; mitomycin C;
Cisplatin derivatives, etc. The compounds of the present invention and their salts can be administered simultaneously with, before or after the anticancer drug, in combination with the anticancer drug, or separately. That is, the compounds of the present invention and their salts can be prepared alone in preparations according to various administration methods, and can be administered separately with various anticancer drugs. It can also be administered after making a preparation according to the administration method. The administration method varies depending on the symptoms of the subject, the properties of the anticancer drug, etc., but the dosage is 1 to 1 times per day for adults.
Divide 1000mg into one or several doses for oral administration as tablets, granules, powders, suspensions, capsules, syrups, etc., or parenteral administration as injections, suppositories, isotonic solutions for infusion, etc. It can be administered as a drug. For example, in the case of tablets, crystalline cellulose, light anhydrous silicic acid, etc. are used as adsorbents, and corn starch, lactose, calcium phosphate, magnesium stearate, etc. are used as excipients. In addition, when preparing an injection, an aqueous solution of the compound or a suspension aqueous solution using cottonseed oil, corn oil, peanut oil, olive oil, etc.
It is used as an emulsion using surfactants such as HCO-60. In addition, as for the administration method of the anticancer drug, various administration methods selected for each anticancer drug can be used as they are. [Effects of the Invention] The compounds of the present invention strongly inhibit the outflow of anticancer drugs from cancer cells, have low toxicity, and have very few side effects such as lowering of blood pressure. For example, for the compound shown in Example 1, in an acute toxicity test using CDF ₁ mice, the LD ₅₀ was
It has low toxicity at over 600mg/Kg, and in a cardiovascular effect test using anesthetized dogs, there was little to no drop in blood pressure or increase in heart rate when 0.1 to 1mg/Kg of the sample was administered intravenously. The effect was minor. Therefore, the compound of the present invention is effective against cancer cells that are less sensitive to anticancer drugs and cancer cells that have acquired resistance to anticancer drugs, and provides a new treatment method for cancer chemotherapy, which is currently at a dead end. It's watery. [Examples] Next, the present invention will be explained in more detail in Examples. Example 1 4-{3-(4-diphenylmethylpiperazine-
1-yl)-2-hydroxypropoxy}-1H
-Indole a 2 g of 4-hydroxyindole, 2.1 g of epichlorohydrin and 3.1 g of potassium carbonate were mixed with 20 ml of acetone and heated under reflux for 6 hours. After filtering off the insoluble matter, the solvent was distilled off, and the residue was purified by silica gel column chromatography. When eluted with chloroform:methanol=100:1, 1.9 g of the target product, 4-(2,3-epoxy-propoxy)-1H-indole, was obtained as an oily substance. IRνcm ^-1 (1iq.f.): 3400, 3000, 1700, 1620,
1590, 1510, 1360 b 0.5 g of the epoxy compound obtained in step a above and 0.66 g of N-diphenylmethylpiperazine were added to 20 ml of ethanol and heated under reflux for 2 hours. The solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography. When chloroform: methanol = 100:1 flows out, the target substance 4
-{3-(4-diphenylmethylpiperazine-1
-yl)-2-hydroxypropoxy}-1H-
0.85 g of indole was obtained as an amorphous powder. IRνcm ^-1 (KBr): 3420, 2920, 2820, 1620,
1580, 1500, 1450, 1360 NMR δppm ( _CDCl3 ) 2.2~2.8 (m, 10H), 3.3 (br, 1H), 3.9~
4.1 (m, 3H), 4.12 (s, 1H), 6.4~6.6
(m, 2H), 6.8-7.45 (m, 13H), 8.12
(br,7,1H) Example 2 4-{3-(4-benzylpiperazin-1-yl)-2-hydroxypropoxy}-1H-indole 4-(2,3 -epoxy-bropoxy)-1H-indole (1.9 g) and N-benzylpiperazine (1.8 g) in 50 ml of ethanol.
The mixture was dissolved in water and heated under reflux for 2 hours. The solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography. Chloroform: methanol =
If it flows out at a ratio of 100:1, the target material 4-{3-(4
2.7 g of -benzylpiperazin-1-yl)-2-hydroxypropoxy}-1H-indole was obtained as an amorphous powder. IRνcm ^-1 (KBr): 3400, 2920, 2820, 1610,
1580, 1510, 1450, 1360 NMR δppm ( _CDCl3 ) 2.0-2.8 (m, 12H), 3.2-3.7 (m, 3H),
4.05 (s, 3H), 6.3-7.4 (m, 12H), 8.65
(s, 1H) Example 3 4-[3-{4-(9-fluorenyl)-piperazin-1-yl}-2-hydroxypropoxy]-
1H-Indole 1.9 g of 4-(2,3-epoxy-bropoxy)-1H-indole obtained in Example 1 a and 2.5 g of N-(9-fluorenyl)-piperazine were added to 50 ml of ethanol and heated for 2 hours. It refluxed. The solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography. When chloroform: methanol = 100:1 flows out, the target product 4-
[3-{4-(9-fluorenyl)-piperazine-1
2.8 g of -yl}-2-hydroxypropoxy]-1H-indole was obtained as an amorphous powder. IRνcm ^-1 (KBr): 3400, 2920, 2800, 1580,
1500, 1440, 1350, 1240 NMR δppm ( _CDCl3 ) 2.1-2.9 (m, 10H), 3.2 (br, s, 1H),
4.1 (s, 3H), 4.8 (s, 1H), 6.4~6.7 (m,
2H), 6.9-7.8 (m, 11H), 8.2 (s, 1H) Example 4 4-[3-{4-(5-dibenzosuberanyl)-piperazin-1-yl}-2-hydroxypropoxy] -1H-indole 0.92 g of 4-(2,3-epoxy-bropoxy)-1H-indole obtained in Example 1 a and N-
1.35 g of 5-dibenzosuberanyl)-piperazine was mixed with 20 ml of ethanol and heated under reflux for 2 hours.
The solvent was distilled off and the residue was purified by silica gel column chromatography. When flowing out with chloroform:methanol=50:1, the target product 4-[3-{4-(5-dibenzosuberanyl)-
2.05 g of piperazin-1-yl}-2-hydroxypropoxy]-1H-indole was obtained. IRνcm ^-1 (KBr): 3400, 2920, 2800, 1580,
1500, 1450, 1360, 1280, 1240 NMR δppm ( _CDCl3 ) 2.0~3.1 (m, 12H), 3.5 (s, 1H), 3.8~
4.3 (m, 6H), 6.4~6.7 (m, 2H), 6.8~
7.4 (m, 11H), 8.25 (s, 1H) Example 5 4-{3-(4-diphenylmethylpiperazine-
1-yl)-2-hydroxypropoxy}-1-
Methylindole Example 1 4.4 g of the compound was dissolved in 50 ml of dry tetrahydrofuran, and 0.4 g of 60% sodium hydride was slowly added. After stirring at room temperature for 1 hour, 10 ml of dry tetrahydrofuran in which 1.42 g of methyl iodide was dissolved was gradually added dropwise. After stirring at room temperature for 6 hours, the solvent was distilled off under reduced pressure, and the residue was dissolved in chloroform and washed with water. The chloroform layer was dried over anhydrous sodium sulfate and then distilled off, and the residue was purified by silica gel column chromatography. When chloroform:methanol=50:1 flows out, the target product 4-{3-(4-diphenylmethylpiperazin-1-yl)-2-hydroxypropoxy}-1-
3.8 g of methyl indole was obtained as an amorphous powder. IRνcm ^-1 (KBr): 3420, 2920, 2800, 1600,
1575, 1490, 1450, 1250, 1050 NMR δppm ( _CDCl3 ) 2.0-2.9 (m, 10H), 2.9-3.4 (br, s,
1H), 3.7 (s, 3H), 3.9-4.4 (d, 4H),
6.4-6.7 (m, 2H), 6.8-7.6 (m, 13H) Example 6 4-{3-(4-diphenylmethylpiperazine-
1-yl)-2-hydroxypropoxy}-1-
Benzoylindole Example 1 4.4 g of the compound was dissolved in 50 ml of dry tetrahydrofuran, and 0.4 g of 60% sodium hydride was slowly added. After stirring for 1 hour at room temperature, 10 ml of dry tetrahydrofuran containing 1.4 g of benzoyl chloride was slowly added dropwise. The reaction solution was heated at room temperature for 5 minutes.
After stirring for an hour, the solvent was distilled off, and the residue was dissolved in chloroform and washed with water. The chloroform layer was dried over anhydrous sodium sulfate and then distilled off, and the residue was purified by silica gel column chromatography. When chloroform:methanol=50:1 flows out, the target product 4-{3-(4-diphenylmethylpiperazin-1-yl)-2-hydroxypropoxy}-
1-benzoylindole as amorphous powder
3.2g was obtained. IRνcm ^-1 (KBr): 2900, 2800, 1680, 1620,
1490, 1445, 1430, 1340, 1270 NMR δppm ( _CDCl3 ) 2.1-3.1 (m, 12H), 3.1-3.6 (m, 1H),
3.9~4.3 (m, 3H), 6.5~6.8 (m, 2H),
6.9-8.1 (m, 18H) Example 7 4-{3-(4-diphenylmethylhomopiperazin-1-yl)-2-hydroxypropoxy}-
1H-Indole 1.9 g of 4-(2,3-epoxy-bropoxy)-1H-indole obtained in Example 1 a and 2.7 g of N-diphenylmethylhomopiperazine were dissolved in 50 ml of ethanol and heated for 2 hours. It refluxed. The solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography. When chloroform:methanol=100:1 flows out, the target product 4-{3-(4-diphenylmethylhomopiperazin-1-yl)-2-hydroxypropoxy}-
2.8 g of 1H-indole was obtained as an amorphous powder. IRνcm ^-1 (KBr): 3400, 2920, 2820, 1610,
1585, 1510, 1500, 1450, 1360 NMR δppm (CDCl ₃ ) 1.7 (t, 3H), 2.4-3.1 (m, 10H), 3.65
(s, 1H), 4.1 (s, 3H), 4,35 (s,
1H), 6.4-6.7 (m, 2H), 6.8-7.6 (m,
13H), 8.25 (s, 1H) Test Example 1 Effect of enhancing anticancer drug retention in drug-resistant cancer cells Adriamycin-resistant strain 2780AD of human ovarian cancer cell A2780 (AM Rogan et al., Science, Vol. 224, 994)
-996 pages, 1984) containing 5% fetal bovine serum.
Suspend 1×10 ⁶ cells/ml in RPMI-1640 culture medium,
1 in a 24-well multi-well culture plate with a diameter of 16 cm.
Seed 1 ml of cancer cell suspension per well and add 5%
Cultured at 37°C in CO ₂ . After 24 hours, remove the culture medium.
20nM ³ H-vincristine (1×10 ⁴ dpm/
pmol), 5% fetal bovine serum, and 10 nM Hepes buffer.
After dissolving in DMSO, 5μ of the test compound diluted with physiological phosphate buffer was added (the concentration in the reaction solution was 1.0
After continuing to incubate for 2 hours at 37° C. or 10.0 μg/ml) and 5% CO ₂ , the cells were washed with cold physiological phosphate buffer. Add 0.5ml of 0.2N-NaOH to this,
The cells were transferred to a vial and bathed at 56°C for 30 to 60 minutes to lyse the cells. 4ml of Acid Aquasol 2
In addition, the amount of ³ H-vincristine in the cells was measured using a liquid scintillation counter. The effect is expressed as a percentage (%) of the amount of vincristine retained in the drug-treated group, with the amount of vincristine retained in the non-drug-treated control group as 100.
It was expressed as The results are shown in Table 1. Test Example 2 Effect of enhancing the action of anticancer drugs Adriamycin-resistant strain K562/ADM of human myeloid leukemia K562 in RPMI containing 5% fetal bovine serum
-1640 culture medium with 2 x ¹⁰⁴ cells/ml suspended, 12 x 75
2 ml of cancer cell suspension per tube was seeded into mm tubes and cultured at 37° C. in 5% CO ₂ . After 6 hours, vincristine (0 to 3000 ng/ml) and the test compound were added to the reaction mixture at concentrations of 0.3, 1, and 3 μg/ml, and the culture was continued at 37° C. and 5% CO ₂ for 72 hours. The cell suspension was added to 9.5 ml ISOTON, and the number of cells was counted using a Coulter counter, and then the 50% growth inhibition concentration IC ₅₀ (ng/ml) of vincristine was determined. The results of tests conducted on the compounds listed in Table 1 are shown in Table 2 in terms of IC ₅₀ values and degree of effect enhancement (multiply: shown in parentheses in Table 2). Other compounds also showed similar action-enhancing effects.

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Claims (1)

[Claims] 1 General formula () (In the formula, R ₁ represents a hydrogen atom, a lower alkyl group, or an acyl group, and R ₂ represents a benzyl group, a diphenylmethyl group, a fluorenyl group, or a dibenzosuberanyl group. n represents 2 or 3.) Compounds represented and their salts. 2 formulas A compound represented by and its salt. 3 formulas A compound represented by and its salt. 4. An anticancer drug effect enhancer comprising a compound represented by the general formula () or a salt thereof as an active ingredient.