JPH02121966A - Indole derivative and effect-enhancing agent for anticancer agent containing same derivative as active ingredient - Google Patents

Abstract

NEW MATERIAL:A compound expressed by formula I and salt of the same compound (R1 is H, lower alkyl or acyl; R2 is benzyl, diphenylmethyl, fluorenyl or dibenzosuberanyl; n is 2 or 3). EXAMPLE:4-[3-(4-diphenylmethylpiperazine-1-yl)-2-hydroxypropoxy]-1H-in dole. USE:Effect-enhancing agent for anticancer drug exhibiting strong effect-enhancing activity for anticancer in resistance cancer and having low toxicity and low adverse effect. PREPARATION:A compound expressed by formula II is reacted with a compound expressed by formula III (X is halogen) in the presence of base and preferably at a temperature of 0-100 deg.C to obtain a compound expressed by formula IV, then resultant compound is reacted with a compound expressed by formula V at room temperature-boiling point of solvent (organic solvent such as alcohol, acetone, chloroform or DMF) to afford the compound expressed by formula I.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application 1] 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 is the highest in our country.1 Social interest in cancer treatment is high.

Research and development of anticancer drugs for the treatment of cancer has been actively conducted for a long time, and clinically, the most promising anticancer drugs are used for the treatment of cancer. Although its effectiveness is steadily improving year by year, in many cases it is not always satisfactory to completely suppress cancer growth and maintain the survival of cancer patients for a long period of time. At present, many attempts are being made clinically to enhance the effects of anticancer drugs by combining 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 a multilayered anticancer drug or a method for more selectively transporting the anticancer drug to the target organ. These studies are currently being carried out all over the world, but the current situation is that they are becoming increasingly difficult.

-51 Another important method is to try to enhance the effects of existing anticancer drugs. In particular, we believe that the development of agents that enhance the effects of existing anticancer drugs against drug-resistant cancer, which is a serious clinical problem in cancer treatment methods, is a very important new cancer treatment method. The background of clinical resistance to anticancer drugs is not necessarily simple; clinical resistance can be roughly 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 has been elucidated at the molecular level.
Treatment methods for this problem are also being studied. In other words, 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).
-glycopr.

It was revealed that the gene was derived from ``Lein''. 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, this P-
Due to the action of glycoproteins, the anticancer drug that has entered the body is expelled from the cell, keeping the concentration of the drug inside the cancer cell 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 an effect of enhancing the effect of anticancer drugs, and is particularly effective in overcoming resistance.
We believe that it can be used as a new cancer chemotherapeutic agent.

In fact, Tsuruo et al. reported that calcium antagonists such as verapamil block the efflux of anticancer drugs from cancer cells, and therefore can be used in combination against resistant cancers in vitro and in vivo.
It has been found that it has the effect of enhancing the effects of anticancer drugs such as adriamycin and vincristine. but,
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 treatment agent. Therefore, there has been a desire for a drug that has a stronger effect of enhancing anticancer drug effects on resistant cancers and has fewer side effects.

[Means for Solving the Problems] Based on the above-mentioned viewpoint, the inventors of the present invention have conducted extensive studies and have found that:
The present invention was completed based on the discovery that the indole derivative shown below exhibits a strong anticancer drug effect enhancing effect on resistant cancers, and has low toxicity and low side effects.

That is, the present invention relates to the general formula (I) (wherein R1 represents a hydrogen atom, a lower alkyl group, or an acyl group, and R2 represents a benzyl group, a diphenylmethyl group, a fluorenyl group, or a dibenzosuberanyl group, n represents 2 or 3) and its salt (hereinafter referred to as 1 compound of the present invention) j5 and an anticancer drug effect enhancer containing them as active ingredients.

The lower alkyl group in general formula (I) is a methyl group,
It refers to an ethyl group, a propyl group, a butyl group, etc., and an acyl group refers to an acetyl group, a propionyl group, a benzoyl group, etc.

Examples of the salts of the compounds of the present invention include salts with inorganic acids such as hydrochloric acid and sulfuric acid, or organic acids such as acetic acid, oxalic acid, maleic acid, and tartaric acid. Further, since the compound of the present invention has an asymmetric carbon in its structure, optical isomers exist, and one compound of the present invention is assumed to contain all of these.

Compounds structurally similar to the indole derivatives of the compounds of the present invention include JP-A-56-32455.57-146754.
There is a group of compounds described in 6137765 and the like. However, in these documents, there are no examples of the compound of the present invention itself. Furthermore, regarding the compound shown in claim 2,
There are no descriptions that define this compound, such as specific physical property values, in Examples, etc., and there is no description of this compound in any other literature, and the above literature does not describe its use in cardiovascular drugs, especially cardiotonic drugs. However, the anticancer drug effect enhancer described in the present invention is not described at all.

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 shrimp bromohydrin in the presence of a base. (Same as above, X represents halogen) leads to the corresponding epoxy compound. Bases here include sodium hydroxide, sodium hydride,
Inorganic bases such as potassium carbonate, t-butoxypotassium, and sodium carbonate 1; organic bases such as triethylamine and pyridine; as the solvent, water solvents or organic heating media such as alcohol, acetone, THF, and DMF can be used. The reaction temperature is preferably in the range of 0 to 00°C.

Furthermore, the compound of general formula (I) of the present invention can be obtained by thermally reacting a corresponding epoxy compound and a corresponding amine derivative.

(In the formula, R1, R2, 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, DMF, etc. of organic solvents are used.

The anticancer drug effect enhancement effect of the compound of the present invention against resistant cancer is as follows:
Adriamycin-resistant human ovarian cancer cell line 2780AD
Alternatively, 562/ADM is used in an adriamycin-resistant strain of human myeloid leukemia cells, and evidenced by the effect of enhancing the retention of the anticancer drug in the cells and the effect of enhancing the action of the anticancer drug.

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.

In addition, the anticancer agent to which the compound of the present invention or a salt thereof is used is not particularly limited, but preferable ones include non-antimetabolite anthracycline antibiotics such as adriamycin, downmycin, and aclacinomycin A: Actinomycin antibiotics, such as actinomycin C, actinomycin D: Chromomycin antibiotics, such as mithramycin, toyomycin: Vinca alkaloids, such as vincustine, vinplastine: Maytansines: Podophyllotoxin derivatives, such as VP
16-213: homohalintonin: anguwidein: purceantin: neocarzinostatin: anthramycin: mitomycin C: cisplatin derivatives.

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. In other words, the compound of the present invention and its salt 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 will naturally depend on the symptoms of the subject, the properties of the anticancer drug, etc., but 1 to 1000 mg per day for adults.
tablets, granules, powders, suspensions,
It can be administered orally, such as capsules and syrups, or parenterally, such as injections, suppositories, and isotonic solutions for infusion.

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 used as an injection,
It is used as an aqueous solution of the compound, a suspension aqueous solution using cottonseed oil, corn oil, peanut oil, olive oil, etc., or an emulsion using a surfactant 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.

[Effect of the Invention 1] The compound of the present invention strongly inhibits the outflow of anticancer drugs from cancer cells, and has the characteristics of low toxicity and 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 and mice, LD showed low toxicity at 600 mg/kg or more when administered i.p. In the effect test, when 0.1-1 mg/kg of the specimen was administered i and v, there was almost no decrease in blood pressure, increase in heart rate, etc., or very slight effects.

Therefore, the compounds of the present invention are effective against cancer cells that are less sensitive to anticancer drugs and cancer cells that have acquired resistance to anticancer drugs, and provide a new treatment method for cancer chemotherapy, which is currently at a dead end.

[Example] Next, the present invention will be explained in more detail in Examples.

Example 1 4-(3-(4-diphenylmethylbiperazinl-yl)-2-hydroxypropoxy)-IH-indole al 2 g of 4-hydroxyindole, 2.1 g of epichlorohydrin and 3.1 g of potassium carbonate were The mixture was mixed with acetone 20IIIf2 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 flowing out with chloroform:methanol=100:l, the target product 4-(2
, 3-epoxy-propoxy)-1-day-indole was obtained as an oil in an amount of 1.9 g.

IRycm-Mliq, f, l: 3400. 30
00. 1700. 1620°1590, 1510.
1360 b) 0.5 g of the epoxy compound obtained in a) above and 0.66 g of N-diphenylmethylpiperazine were added to ethanol 20I2, and the mixture was 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, 0.85 g of the target product 4-13-(4-diphenylmethylpiperazin-1-yl)-2-hydroxypropoxy)-IH-indole is obtained as an amorphous powder.
Obtained 6IR1/cm-' (KBr): 3420.2920
．． 2820.1620°1580, 1500.1450
．． 136ONMRδppm (CDC1a) 2.2-2.8 (m, 10H), : 1.3 (br, I
Hl, 3.9-4, lfm, 3H1,4,12(s,
IH), 6.4~6.6 (!1.2H1゜6.8~7
．． 45(m, 13H1,8,12(br,s,lH) Example 2 4-(3-(4-benzylpiperazin-1-yl)-
2-Hydroxypropoxy)-IH-indole 1.9 g of 4-(2,3-epoxy-propoxy)-1)]-indole obtained according to a) of Example 1 and 18 g of N-benzylpiperazine were dissolved in 50 mβ of ethanol. The mixture was dissolved 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.

The target product 4-(3-(4-benzylpiperazine-1-
2.7 g of (2-hydroxypropoxy)-IH-indole was obtained as an amorphous powder.

IR1/cn+-'fKBr): 3400.292
0.2820.1610.15801510, 145
0. 136O N14Rδppn+ (CDC13) 2.0~2.8(m, 12H1,3,2~3.7fm,
3H1, 4, 05fs, 3H), 6.3~? , 4f
m, 12H1,8,65fs, lH) Example 3 4-[3-(4-(9-fluorenyl)-piperazin-1-yl)-2-hydroxypropoxy]-IH-indole according to a) of Example 1 1.9 g of the obtained 4-(2,3-epoxy-propoxy)-LH-indole and N-(9
-fluorenyl)-piperazine 2.5 g ethanol 5
0II112 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 = 100.1
When the mixture was flowed out, 2.8 g of the target product 4-[3-(4-(9-fluorenyl)-piperazin-1-yl)-2-hydroxypropoxy]-1F-1-indole was obtained as an amorphous powder. .

IRvam-' (Br): 3400.2920
．． 2800.1580.15001440, 1355.
124O NMR δppn+ (CDC131 2.1-2.9fm, 1OH), 3.2(br, s
, IH), 4. N53H), 4.8 (s, l old,
6.4-6.7fm, 2H1, 6, 9-7, Hm,
111(I,8,Hs,1J(lExample 4 4-[3-(4-(5-dibenzosuberanyl)-piperazin-1-yl)-2-hydroxypropoxy]-
LH-indole 0.92 g of 4-(2,3-epoxy-propoxy)-18-indole obtained in Example 1 a) and N-(5-
1.35 g of dibenzosuberanyl)-piperazine was mixed with 20 n112 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:l, the target product 4-[3-(4
-(5-dibenzosuberanyl)-piperazin-1-yl)-2-hydroxypropoxy] 2.05 g of -18-indole was obtained.

IRvcm-'(KBr): 3400.2920.
2800.1580.15001450, 1360.1
280.124ONMRδ pI)II fcDc1
3) 2.0-3.1 (water, 12H1,3,5(s, lH
l, 3.8-4.3 (m.

6H1, 6, 4~6.7fn+, 2H), 6.8~7
．． 4 (m, 1lH).

8, 25 fs, IHI Example 5 4-(3-(4-diphenylmethylbiperazine-1-
yl)-2-hydroxypropoxy)-1-methylindole Example 1 4.4 g of compound was dissolved in 50 g of dry tetrahydrofuran.
mβ and 0.4 g of 60% sodium hydride was gradually added. After stirring for 1 hour at room temperature, 1Of dry tetrahydrofuran in which 1.42 g of methyl iodide was dissolved.
1112 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.
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-diphenylmethylbiperazin-1-yl)-2-hydroxypropoxy)-1-methylindole is dissolved as an amorphous powder. .8g
Obtained.

IRvcm-'(Brl: 3420.2920
．． 2800.1600.1575+490.1450.
1250.1050NMRδppm fcoct31 2.0-2.9 (m, old IH, 2.9-3.4 (br,
s, IHl: 1.7(s, 3H1,3,9~4.4(d
, 4H1,6,4-6.7 (m.

2H1,6,8-7.6fm, 13 Old Example 6 4- (3-(4-diphenylmethylbiperazine-1-
yl)-2-hydroxypropoxy)-1-benzoylindole Example 1 4.4 g of compound was dissolved in 50 g of dry tetrahydrofuran.
+nJ2 and 0.4 g of 60% sodium hydride was gradually added. After stirring for 1 hour at room temperature, 1.4 g of benzoyl chloride in dry tetrahydrofuran was added.
Omβ was gradually added dropwise. After stirring the reaction solution at room temperature for 5 hours, the solvent was distilled off, and the residue was dissolved in chloroform.
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-diphenylmethylbiperazin-1-yl)-2-hydroxypropoxy 1-1-benzoylindole) is dissolved as an amorphous powder. ．．
2g was obtained.

lRv cm-' (Brl: 2900.280
0.1G80.1620.14901445, 1430
．． 1340.127ONMRδppm (CDC1+1 2.1~3.1(m, 12 old, 3.1~3.6(m,
IH), 3.9-4.3in+, 311), 6.
5-6.8fm, 2H), 6.9-8. Nm.

8H1 Example 7 4-(3-(4-diphenylmethylhomopiperazine-
1.9 g of 4-(2,3-epoxy-propoxy)-18-indole obtained according to a) of Example 1 and N-diphenylmethylhomopiperazine 2 .7g and ethanol5
The solution was dissolved in 0ml 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 = 100:
1, the target product 4-(3-(4-diphenylmethylhomopiperazin-1-yl)-2-hydroxypropoxy 1-IH-indole) is released as an amorphous powder.
．． 8g was obtained.

IRvcm-'(KBrl: 3400.2920
．． 2820.1610.15851510, 1500
．． 1450. 136ONMRδppm f(:D
C13) 1.7it, 3H1,2,4~3.1fm, 1OH),
3.651s, LHl.

4, l (s, 3H), 4.35 (s, l
Old, 6.4-6.7 (m, 2 old.

6.8-7.6 (m, 13H1, 8, 25fs, IH)
Test example 1. Effect of enhancing anticancer drug retention in drug-resistant cancer cells Adriamycin-resistant strain 2 of human ovarian cancer cell A2780
780 A D (A, M. Logan et al., 5cie
nce, vol. 224, pp. 994-996, 1984
2000) was suspended in RPMI-1640 culture medium containing 5% tallow serum and 1 ml per well was placed in a 24-sized multi-well culture plate with a diameter of 16 cm.
A suspension of cancer cells was seeded and cultured at 37°C in 5% CO*. After 24 hours, the culture solution was mixed with RPMI-1 containing 20 nM'H-vincristine (I x lo'dpm/pmol), 5% tallow serum, and 10 nM Hepes buffer.
It was replaced with 0.5 nl of 640 culture solution. After dissolving in DMSO, the test compound diluted in physiological phosphate buffer was added to 5 μβ
(Concentration in reaction solution is 1.0 or IO, 0 μg/+
nl2), After continuing to culture for 2 hours at 37°C in 5% CO2, the cells were washed with chilled physiological phosphate buffer. 0.5 mI2 of 0.2 N-Na011 was added thereto, transferred to a vial, and bathed at 56°C for 30 to 60 minutes to lyse the cells. Add 4mβ of Acid Aquasol 2,
The amount of 3H-vincristine in the cells was measured using a liquid scintillation counter.

The effect was expressed as a percentage (%) of the amount of vincristine retained in the drug-treated group, with the amount of vincristine retained in the drug-untreated control group taken as 100. The results are shown in Table 1.

Test example 2. RPMI-16 containing 5% tallow serum containing 562/ADM on adriamycin-resistant strains of 562 on human myeloid leukemia
Suspend 2 x 10' cells in 7 ml of 40 culture medium,
A cancer cell suspension was seeded at 2m°C per tube in x75mm tubes, and cultured at 37°C in 5% CO□. 6
After hours, vincristine (0-3.000 ng/mf
f) and the test compound at a concentration of 03, 1, 3 gg during the reaction.
/mff and 72°C at 37°C with 5% CO□.
Culture was continued for hours. Cell suspension at 9.5mI21SOT
In addition to ONII, after counting the number of cells with a Coulter counter, the 50% growth inhibitory concentration of vincristine I Cso
(ng/1tj2) was determined.

Table 2 shows the results of tests conducted on the compounds listed in Table 1.
Cs. Value and effect enhancement degree (multiple: shown in parentheses in Table 2)
Indicated by Other compounds also showed similar action-enhancing effects.

Table 1 Procedure Nefu Seisho (White 2 February 7, 1999

Claims (1)

[Claims] 1. General formula (I) ▲ Numerical formulas, chemical formulas, tables, etc. ▼ (I) (In the formula, R_1 represents a hydrogen atom, a lower alkyl group, or an acyl group, and R_2 represents a benzyl group or diphenyl group. (a methyl group, a fluorenyl group, and a dibenzosuberanyl group; n represents 2 or 3) and salts thereof. 2. Compounds and their salts represented by the formula ▲There are mathematical formulas, chemical formulas, tables, etc.▼. 3. Compounds represented by the formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ and their salts. 4. An anticancer drug effect enhancer containing a compound represented by general formula (I) or a salt thereof as an active ingredient.