JPH0920778A - Pyrazoloindolium derivative - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new pyrazoloindolium derivative highly inhibiting topoisomerase I and topoisomerase II, having proliferation-inhibitory activity on oncocytes, thus useful as an antineoplastic agent. SOLUTION: This compound is expressed by formula I [Q is CH or N; R<1> and R<2> are H, a halogen, 1-6C alkyl(amino), etc.; R<3> is a (halogen etc.-substituted) 1-6C alkyl; R<4> is H, a (halogen etc.-substituted) 1-6C alkyl, (halogen etc.,- substituted)-phenyl, (halogen etc.,-substituted) phenyl-substituted 1-6C alkyl, etc.; R<5> is the same as R<4> except H; X<-> is a base-forming anion and may be absent when one of substituents R<1> to R<5> is a proton-donating group], e.g. 1- methyl-4-(Z-(p-dimethylaminophenylmethylene)-2-phenyl-4H-pyrazolo[1,5- a]indolium trifluoromethanesulfonate. The compound of formula I is obtained by oxidizing and quaternizing a compound of formula II to obtain a compound of formula III, which is then condensed with a compound of the formula R<5> -CHO.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel compound having an antitumor activity and a medicine containing the compound as an active ingredient. More specifically, the present invention relates to a pyrazoloindolium derivative having an exomethylene group or an imino group and having an antitumor effect, and an antitumor agent containing the compound as an active ingredient.

[0002]

Various antitumor compounds are clinically used as cancer chemotherapeutic agents. However, antitumor compounds generally have strong cytotoxicity against normal cells, and it is known that nausea, hair loss, leukopenia, etc. occur during cancer chemotherapy. It is also known that tumor cells acquire resistance to antitumor agents. Under such circumstances, development of a cancer chemotherapeutic drug based on a new mechanism of action is required.

DNA topoisomerase is an enzyme that catalyzes the conversion of a higher-order structure (supercoil) of DNA, and is known to be an enzyme essential for cell growth accompanied by a DNA replication process in eukaryotes and prokaryotes. There is. Eukaryotic cells and prokaryotic cells have two types of DNA topoisomerases, namely topoisomerase I,
Type I topoisomerases: Topo I) and topoisomerases
II (topoisomerase II, type II topoisomerase: Topo I
The existence of I) is clarified. Prokaryotic topoisomerase II is sometimes called gyrase.

The functions of these two enzymes in eukaryotes have not been fully elucidated, but topoisomerase II
Is the action of forming an energy-labile negative supercoiled DNA from double-stranded DNA and supercoiled DNA.
Has the effect of forming relaxed DNA that is energetically stable from ATP.
It is known that the energy supply from In addition, topoisomerase I is a supercoiled DN
It has the effect of forming relaxed DNA from A, and this effect has also been revealed to be non-auxotrophic for ATP and Mg ²⁺ ions.

[0005] Among substances that inhibit the action of DNA topoisomerase existing in mammals, it is known that certain substances show a selective growth inhibitory action on mammalian tumor cells and are useful as antitumor agents. Has been. For example, topotesin, which inhibits topoisomerase I, and etoposide, which inhibits topoisomerase II (Topo II), are used clinically. However, topoisomerase
A substance having an inhibitory effect on both I and topoisomerase II has not yet been developed as an antitumor agent.
A substance having such an action does not show cross resistance even to a tumor cell that has been made resistant to an existing antitumor agent, and can be expected to be useful as an antitumor agent.

On the other hand, compounds having a pyrazoloindole skeleton are described, for example, in Katayama et al. (Katayama, H., et al., Chem.
Pharm. Bull., 38, pp. 1129-1135, 1990); Katayama et al. (Kata
yama, H., et al., Chem. Pharm. Bull., 40, pp.2267-
2269, 1992); Shen et al. (Shen, JK, et al., J. Chem.
Soc. Perkin Trans.1, pp.2087-2097, 1993); and Shen et al. (Shen, JK, et al., J. Chem. Soc. Perkin Tr.
ans.1, pp.1871-1877,1994), but these publications do not suggest or teach the DNA topoisomerase inhibitory action of compounds having a pyrazoloindole skeleton.

[0007]

DISCLOSURE OF THE INVENTION The object of the present invention is to provide DNA
It is to provide a novel compound having an inhibitory effect on topoisomerase, and specifically to provide a novel compound having an inhibitory effect on both topoisomerase I and topoisomerase II. Also, the present invention
It is an object of the present invention to provide a compound having a DNA topoisomerase inhibitory action and useful as an active ingredient of an antitumor agent, and further topoisomerase I and topoisomerase.
It is also an object of the present invention to provide a compound having an inhibitory effect on both of II and useful as an active ingredient of an antitumor agent.

[0008]

The inventors of the present invention have made diligent efforts to solve the above-mentioned problems, and as a result, a novel pyrazoloindolium derivative represented by the following general formula (I) has been identified as topoisomerase I and topoisomerase II. It was found that they have strong inhibitory activity against any of them and are extremely useful as an active ingredient of an antitumor agent based on a new mechanism of action. The present invention has been completed based on the above findings.

That is, the present invention has the following general formula (I):

Embedded image [In the formula, Q represents CH or a nitrogen atom; R ¹ and R ² are each independently a hydrogen atom, a halogen atom, a C _1-6 alkyl group,
Amino group, C _1-6 alkylamino group, C _1-6 alkylcarbonylamino group, hydroxy group, C _1-6 alkoxy group, thiol group, C _1-6 alkylthio group, cyano group, carbamoyl group, nitro group, acyl A group or an acyloxy group;
R ³ is a C _1-6 alkyl group (C _1-6 alkyl group is a halogen atom, amino group, C _1-6 alkylamino group, C _1-6 alkylcarbonylamino group, hydroxy group, C _1-6 alkoxy group ,
A thiol group, a C _1-6 alkylthio group, a cyano group, a carbamoyl group, a nitro group, an acyl group and an acyloxy group, which may have one or more substituents); R ⁴ is a hydrogen atom, a C _1-6 alkyl group, a phenyl group, a phenyl-substituted C _1-6 alkyl group, or a phenyl-substituted C _2-6 alkenyl group (C _1-6 alkyl group, phenyl group,
The phenyl group portion of the phenyl-substituted C _1-6 alkyl group, or the phenyl group portion of the phenyl-substituted C _2-6 alkenyl group is a halogen atom, a C _1-6 alkyl group, an amino group, a C _1-6 alkylamino group, C _1-6 alkylcarbonylamino group, hydroxy group, C _1-6 alkoxy group, thiol group, C _1-6 alkylthio group, cyano group, carbamoyl group, nitro group, acyl group,
And a substituent selected from the group consisting of acyloxy groups
Or may have two or more); R ⁵ is a C _1-6 alkyl group, a phenyl group, a phenyl-substituted C _1-6 alkyl group or a phenyl-substituted C _2-6 alkenyl group (C _1-6 The alkyl group, the phenyl group, the phenyl group portion of the phenyl-substituted C _1-6 alkyl group, or the phenyl group portion of the phenyl-substituted C _2-6 alkenyl group is a halogen atom, a C _1-6 alkyl group, an amino group, C _{1- 6}
Alkylamino group, C _1-6 alkylcarbonylamino group,
Hydroxy group, C _1-6 alkoxy group, thiol group, C _1-6 alkylthio group, cyano group, carbamoyl group, nitro group,
1 or 2 or more substituents selected from the group consisting of an acyl group and an acyloxy group are shown);
X ⁻ represents a salt-forming anion (provided that any one of the substituents R ¹ to R ⁵ is a proton donating group, X ⁻ does not need to be present). To provide various compounds.

According to another aspect of the present invention, a DNA topoisomerase inhibitor comprising the above compound; a DNA topoisomerase inhibitor for eukaryotic cells comprising the above compound; topoisomerase I and topoisomerase II inhibitors comprising the above compounds; Eukaryotic topoisomerase I and topoisomerase II inhibitors consisting of compounds; medicines consisting of the above compounds; medicines containing the above compounds as active ingredients; each of the above medicines used as antitumor agents; pharmaceutical compositions containing the above compounds as active ingredients And a pharmaceutical composition as described above for use as an antitumor agent.

[0011]

BEST MODE FOR CARRYING OUT THE INVENTION In the present specification, an alkyl group,
As the alkoxy group and the alkenyl group, either a straight chain or a branched chain may be used. The alkenyl group may have one or more unsaturated bonds, and each unsaturated bond is an E-
Or in the Z-configuration. Also, for example, C
_When described as _1-6 alkyl group, it means an alkyl group having 1 to 6 carbon atoms. As the C _1-6 alkyl group, for example, methyl group, ethyl group, n-propyl group, isopropyl group, n-
A butyl group, sec-butyl group, tert-butyl group, n-pentyl group, n-hexyl group and the like can be used, and as the C _1-6 alkoxy group, for example, a methoxy group, an ethoxy group, n-
Propoxy group, isopropoxy group, n-butoxy group, sec-
A butoxy group, a tert-butoxy group, an n-pentoxy group, an n-hexoxy group and the like can be used. In addition, in the present specification, as the halogen atom, any of a fluorine atom, a chlorine atom, a bromine atom, or an iodine atom may be used.

In the above general formula, R ¹ and R ² are each independently a hydrogen atom, a halogen atom, a C _1-6 alkyl group, an amino group, a C _1-6 alkylamino group, a C _1-6 alkylcarbonylamino group. , Hydroxy group, C _1-6 alkoxy group, thiol group, C _1-6 alkylthio group, cyano group, carbamoyl group,
A nitro group, an acyl group, or an acyloxy group is shown. C _1-6
As the alkylamino group, one C _1-6 alkyl group or an amino group substituted with the same or different two C _1-6 alkyl groups can be used, and examples thereof include a methylamino group, an ethylamino group, A methylethylamino group, a dimethylamino group, an n-propylamino group, an n-butylamino group or the like can be used. As the C _1-6 alkylcarbonylamino group, for example, an acetylamino group, a propionylamino group or the like can be used.

The acyl group (R'-CO-) is an aldehyde group
Any of (R '= hydrogen atom), alkyl-substituted carbonyl group (R' = alkyl group), and aryl-substituted carbonyl group (R '= aryl group) may be used. The alkyl group substituted with a carbonyl group in the alkyl-substituted carbonyl group is preferably C _1-6 alkyl, and the aryl group substituted with a carbonyl group in the aryl-substituted carbonyl group is an aryl group such as a phenyl group or a naphthyl group, preferably a phenyl group Can be mentioned. The acyloxy group means a group (R'-CO-O-) formed by bonding an oxygen atom to the carbonyl carbon of the above acyl group.

R ¹ and R ² can be substituted at any two positions on the 5- to 8-position of the compound represented by the above formula, and the respective substitution positions are not particularly limited. When R ¹ represents a hydrogen atom, R ² is preferably a 6-position substituent. In this case, R ² is an amino group, C _1-6 alkylamino group, hydroxy group, a halogen atom, or more preferably an acyloxy group, R ² is an amino group, hydroxy group,
Most preferably, it is a chlorine atom, a fluorine atom, or an acetoxy group. A compound in which R ¹ and R ² simultaneously represent hydrogen atoms is the most preferred embodiment of the present invention.

R ³ represents a C _1-6 alkyl group which substitutes on the 1-position nitrogen atom to form a quaternary salt. The alkyl group represented by R ³ is a halogen atom, an amino group, a C _1-6 alkylamino group,
C _1-6 alkylcarbonylamino group, hydroxy group, C _1-6
Having one or more substituents selected from the group consisting of an alkoxy group, a thiol group, a C _1-6 alkylthio group, a cyano group, a carbamoyl group, a nitro group, an acyl group, and an acyloxy group at any position May be. When it has two or more substituents, the respective substituents may be the same or different and each may be substituted on the same or different carbon of the C _1-6 alkyl group. For example, as R ³ , an unsubstituted C _1-6 alkyl group, an aralkyl group such as a benzyl group or a phenethyl group, a trifluoromethyl group or 2,2,2-
A halogenated alkyl group such as a trifluoroethyl group is preferable. Of these, an unsubstituted linear C _1-4 alkyl group is more preferable, and a methyl group is particularly preferable.

R ⁴ is a hydrogen atom, a C _1-6 alkyl group, a phenyl group, a phenyl-substituted C _1-6 alkyl group, or a phenyl-substituted C
_2-6 represents an alkenyl group, R ⁵ is a C _1-6 alkyl group, a phenyl group, a phenyl-substituted C _1-6 alkyl group or a phenyl-substituted C
_2-6 shows an alkenyl group. In these groups, a C _1-6 alkyl group, a phenyl group, a phenyl group portion of a phenyl-substituted C _1-6 alkyl group, or a phenyl group portion of a phenyl-substituted C _2-6 alkenyl group is a halogen atom, C _1-6 Alkyl group, amino group, C _1-6 alkylamino group, C _1-6 alkylcarbonylamino group, hydroxy group, C _1-6 alkoxy group, thiol group, C _1-6 alkylthio group, cyano group, carbamoyl group, nitro It may have one or more substituents selected from the group consisting of a group, an acyl group, and an acyloxy group at any position. When it has two or more substituents, each substituent may be the same or different,
Each can be substituted on the same or different carbon of the C _1-6 alkyl group or on any different carbon of the phenyl ring.

R ⁴ is a hydrogen atom, a C _1-6 alkyl group,
A phenyl group, a phenyl-substituted C _1-6 alkyl group, or a phenyl-substituted C _2-6 alkenyl group is preferable. Further, one substituent selected from a C _1-6 alkylamino group, a hydroxy group, an acyloxy group, or a nitro group may be substituted at any position, preferably
A phenyl group at the p-position can also be preferably used as R ⁴ .
As R ⁴ , more preferably, a hydrogen atom, a linear C _1-4 alkyl group, a phenyl group, a phenyl-substituted linear C _1-4 alkyl group such as a benzyl group or a phenethyl group, a 2-phenylethenyl group or
Phenyl-substituted linear C _2-4 such as 3-phenyl-1-propenyl group
Use an alkenyl group, pC _1-4 dialkylaminophenyl group such as p-dimethylamino group, p-hydroxyphenyl group, pC _1-4 alkylcarbonyloxyphenyl group such as p-acetoxyphenyl group, or p-nitrophenyl group be able to. For example, R ⁴ is preferably a C _1-6 alkyl group, a phenyl group, a phenyl-substituted C _1-6 alkyl group, or a phenyl-substituted C _2-6 alkenyl group, more preferably a hydrogen atom, a methyl group, Phenyl group, phenethyl group and 2-phenylethenyl group, and methyl group, phenyl group and 2-phenylethenyl group are the most preferable groups.

R ⁵ is preferably a phenyl group or a phenyl group having one substituent selected from a C _1-6 alkylamino group, a hydroxy group, an acyloxy group, or a nitro group at the p-position. More preferably, phenyl group, p-aminophenyl group, pC _1-6 alkylaminophenyl group, p-hydroxyphenyl group, p-acyloxyphenyl group, p-nitrophenyl group, pC _1-6 alkoxyphenyl group, or pC _{A 1-6} alkylcarbonylaminophenyl group can be used. More preferably, pC _1-4 mono- or dialkylaminophenyl group such as phenyl group, p-aminophenyl group, p-methylaminophenyl group and p-dimethylaminophenyl group, p-hydroxyphenyl group, acetylaminophenyl group, etc. The pC _1-4 linear alkylcarbonylaminophenyl group, p-nitrophenyl group, pC _1-4 linear alkoxyphenyl group, or pC _1-4 linear alkylcarbonyloxyphenyl group such as acetoxyphenyl group may be used. it can. Particularly preferred groups are phenyl group, p-diC _1-4 linear alkylaminophenyl group, p-nitrophenyl group, p-hydroxyphenyl group, pC _1-4 linear alkylcarbonyloxyphenyl group, and phenyl group , P-dimethylaminophenyl group, p-nitrophenyl group, p-hydroxyphenyl group and p-acetoxyphenyl group are most preferable.

Q represents CH or a nitrogen atom. More specifically, when Q is CH, = QR ⁵ is an exomethylene group (=
CH-R ⁵ ), and when Q is CH, = QR ⁵ is an imino group (= N-
R ⁵ ) is shown. X ⁻ represents a salt-forming anion, and when any one of the substituents R ¹ to R ⁵ is a proton-donating group, the positive charge of the 1-position nitrogen atom and the proton-donating group There is a case where a molecule having a neutral charge (so-called Zwitter ion type molecule) is formed in balance with the negative charge of. Therefore, it goes without saying that a Zwitter-ion type compound having no X ⁻ is also included in the scope of the present invention.
Examples of such a case include the case where R ¹ is a hydroxyl group.

Salt-forming anions are especially those which are capable of pairing with the quaternary ammonium cation formed by the 1-position nitrogen atom to give a molecule of substantially neutral charge. There is no limitation, and for example, a hydroxy anion; a halogen anion such as a chloro anion, a bromo anion, an iodo anion, a fluoro anion; and an acid anion can be used. The acid anion broadly means an atomic group remaining after removing a hydrogen atom from a proton donating molecule, and more specifically, examples thereof include an organic acid residue and an inorganic acid residue.

Examples of the organic acid residue include lower fatty acids such as acetic acid, propionic acid, butyric acid, trifluoroacetic acid and trichloroacetic acid; substituted or unsubstituted benzoic acid such as benzoic acid and p-nitrobenzoic acid; methanesulfone. Acid, (halo) lower alkyl sulfonic acid such as trifluoromethane sulfonic acid; benzene sulfonic acid, p-nitrobenzene sulfonic acid, p-bromobenzene sulfonic acid, toluene sulfonic acid, 2,4,6-triisopropylbenzene sulfonic acid, etc. Substituted or unsubstituted aryl sulfonic acids; and the remaining atomic groups obtained by removing hydrogen atoms from organic phosphoric acid such as diphenyl phosphoric acid. Examples of the inorganic residue include nitrous acid, nitric acid, sulfuric acid or perchloric acid, or a remaining atomic group obtained by removing a hydrogen atom from hydrohalic acid such as borofluoric acid.

When the compound of the present invention has a basic substituent such as an aminophenyl group or a dimethylaminophenyl group, the compound of the present invention may exist in a free form or an acid addition salt form with respect to these groups. is there. In such a case, as the acid addition salt, for example, a salt of the above organic acid or inorganic acid may be used. In the compound represented by the general formula (I), the substituted exomethylene group or the substituted imino group represented by = QR ⁵ has Z-configuration or
Although there are compounds with the E-configuration of the conformation, any of these pure forms of the compounds or mixtures in any proportion are within the scope of the invention. Further, the compound of the present invention may have one or two or more asymmetric carbons depending on the kind of the substituent, but any optical isomer based on one or two or more asymmetric carbons, or 2 Any diastereoisomer based on one or more asymmetric carbons, or a pure form of the above isomer, a mixture of any two or more of the above isomers at any ratio, or a racemate is also within the scope of the present invention. Included. Any hydrate or any solvate of the compounds of the present invention in the various forms described above are also included in the scope of the present invention.

The preferred compounds of the present invention are shown in Table 1 below, but the scope of the present invention is not limited to these compounds. In the table, for example, notations such as 6-NH ₂ and p- (CH ₃ ) ₂ NC ₆ H ₅ indicate an amino group and a p-dimethylaminophenyl group substituted at the 6-position, respectively, and OAc represents an acetoxy group. . TFMS is CF ₃ SO ₃ ^- shows the (trifluoromethanesulfonic acid anion), also exemplified compound numbers in the table corresponds to the compound numbers in the examples, particularly preferred compounds of these compounds the invention Is.

[0024]

[Table 1] ─────────────────────────────────── R ¹ R ² R ³ R ⁴ R ⁵ QX ─────────────────────────────────── HH CH ₃ C ₆ H ₅ C ₆ H ₅ CH TFMS (Example 2) HH CH ₃ C ₆ H ₅ p- (CH ₃ ) ₂ NC ₆ H ₅ CH TFMS (Example 1) HH CH ₃ C ₆ H ₅ p-NO ₂ -C ₆ H ₅ CH TFMS (Example 3) HH CH ₃ C ₆ H ₅ p-OH-C ₆ H ₅ CH TFMS (Example 4) HH CH ₃ C ₆ H ₅ p-OCH ₃ -C ₆ H ₅ CH TFMS HH CH ₃ C ₆ H ₅ p-OAc-C ₆ H ₅ CH TFMS (Example 5) HH CH ₃ C ₆ H ₅ p-NH ₂ -C ₆ H ₅ CH TFMS HH CH ₃ C ₆ H ₅ p-NHCH ₃ -C ₆ H ₅ CH TFMS HH CH ₃ C ₆ H ₅ p-NHAc-C ₆ H ₅ CH TFMS HH CH ₃ C ₆ H ₅ C ₆ H ₅ N TFMS HH CH ₃ C ₆ H ₅ p- (CH ₃ ) ₂ NC ₆ H ₅ N TFMS (Example 8) HH CH ₃ C ₆ H ₅ p-NO ₂ -C ₆ H ₅ N TFMS HH CH ₃ C ₆ H ₅ p-OH-C ₆ H ₅ N TFMS HH CH ₃ C ₆ H ₅ p-OAc-C ₆ H ₅ N TFMS HH CH ₃ C ₆ H ₅ -CH = CH- C ₆ H ₅ CH TFMS HH CH ₃ C ₆ H ₅ -CH ₂ CH ₂ -C ₆ H ₅ CH TFMS HH CH ₃ CH ₃ C ₆ H ₅ CH TFMS HH CH ₃ HC ₆ H ₅ CH TFMS HH CH ₃ p- (CH ₃ ) ₂ NC ₆ H ₅ C ₆ H ₅ CH TFMS HH CH ₃ p-OH-C ₆ H ₅ C ₆ H ₅ CH TFMS HH CH ₃ p-AcO-C ₆ H ₅ C ₆ H ₅ CH TFMS HH CH ₃ p-NO ₂ -C ₆ H ₅ C ₆ H ₅ CH TFMS HH CH ₃ C ₆ H ₅ -CH = CH- p- (CH ₃ ) ₂ NC ₆ H ₅ CH TFMS (Example 6) HH CH ₃ C ₆ H ₅ -CH ₂ CH ₂ -p -(CH ₃ ) ₂ NC ₆ H ₅ CH TFMS HH CH ₃ CH ₃ p- (CH ₃ ) ₂ NC ₆ H ₅ CH TFMS (Example 7) HH CH ₃ H p- (CH ₃ ) ₂ NC ₆ H ₅ CH TFMS HH CH ₃ p- (CH ₃ ) ₂ NC ₆ H ₅ p- (CH ₃ ) ₂ NC ₆ H ₅ CH TFMS HH CH ₃ p-OH-C ₆ H ₅ p- (CH ₃ ) ₂ NC ₆ H ₅ CH TFMS HH CH ₃ p-AcO-C ₆ H ₅ p- (CH ₃ ) ₂ NC ₆ H ₅ CH TFMS HH CH ₃ p-NO ₂ -C ₆ H ₅ p- (CH ₃ ) ₂ NC ₆ H ₅ CH TFMS H 6-NH ₂ CH ₃ C ₆ H ₅ C ₆ H ₅ CH TFMS H 6-NH ₂ CH ₃ C ₆ H ₅ p- (CH ₃ ) ₂ NC ₆ H ₅ CH TFMS H 6-NH ₂ CH ₃ C ₆ H ₅ p-NO ₂ -C ₆ H ₅ CH TFMS H 6-NH ₂ CH ₃ C ₆ H ₅ p-OH-C ₆ H ₅ CH TFMS H 6-NH ₂ CH ₃ C ₆ H ₅ p-OCH ₃ -C ₆ H ₅ CH TFMS H 6-NH ₂ CH ₃ C ₆ H ₅ p-OAc-C ₆ H ₅ CH TFMS H 6-NH ₂ CH ₃ C ₆ H ₅ p-NH ₂ -C ₆ H ₅ CH TFMS H 6-NH ₂ CH ₃ C ₆ H ₅ p-NHCH ₃ -C ₆ H ₅ CH TFMS H 6-NH ₂ CH ₃ C ₆ H ₅ p-NHAc-C ₆ H ₅ CH TFMS H 6-NH ₂ CH ₃ C ₆ H ₅ -CH = CH- C ₆ H ₅ CH TFM S H 6-NH ₂ CH ₃ C ₆ H ₅ -CH ₂ CH ₂ -C ₆ H ₅ CH TFMS H 6-NH ₂ CH ₃ CH ₃ C ₆ H ₅ CH TFMS H 6-NH ₂ CH ₃ HC ₆ H ₅ CH TFMS H 6-NH ₂ CH ₃ p- (CH ₃ ) ₂ NC ₆ H ₅ C ₆ H ₅ CH TFMS H 6-NH ₂ CH ₃ p-OH-C ₆ H ₅ C ₆ H ₅ CH TFMS H 6-NH ₂ CH ₃ p-AcO-C ₆ H ₅ C ₆ H ₅ CH TFMS H 6-NH ₂ CH ₃ p-NO ₂ -C ₆ H ₅ C ₆ H ₅ CH TFMS H 6-NH ₂ CH ₃ C ₆ H ₅ p- (CH ₃ ) ₂ NC ₆ H ₅ N TFMS H 6-NH ₂ CH ₃ C ₆ H ₅ -CH = CH- p- (CH ₃ ) ₂ NC ₆ H ₅ CH TFMS H 6-NH ₂ CH ₃ CH ₃ p- (CH ₃ ) ₂ NC ₆ H ₅ CH TFMS H 6-OH CH ₃ C ₆ H ₅ C ₆ H ₅ CH TFMS H 6-OH CH ₃ C ₆ H ₅ p- (CH ₃ ) ₂ NC ₆ H ₅ CH TFMS H 6-OH CH ₃ C ₆ H ₅ p-NO ₂ -C ₆ H ₅ CH TFMS H 6-OH CH ₃ C ₆ H ₅ p-OH-C ₆ H ₅ CH TFMS H 6-OH CH ₃ C ₆ H ₅ p-OCH ₃ -C ₆ H ₅ CH TFMS H 6-OH CH ₃ C ₆ H ₅ p-OAc-C ₆ H ₅ CH TFMS H 6-OH CH ₃ C ₆ H ₅ p-NH _2- C ₆ H ₅ CH TFMS H 6-OH CH ₃ C ₆ H ₅ p-NHCH ₃ -C ₆ H ₅ CH TFMS H 6-OH CH ₃ C ₆ H ₅ p-NHAc-C ₆ H ₅ CH TFMS H 6- OH CH ₃ C ₆ H ₅ -CH = CH- C ₆ H ₅ CH TFMS H 6-OH CH ₃ C ₆ H ₅ -CH ₂ CH ₂ -C ₆ H ₅ CH TFMS H 6-OH CH ₃ CH ₃ C ₆ H ₅ CH TFMS H 6-OH CH ₃ HC ₆ H ₅ CH TFMS H 6-OH CH ₃ p- (CH ₃ ) ₂ NC ₆ H ₅ C ₆ H ₅ CH TFMS H 6-OH CH ₃ p-OH-C ₆ H ₅ C ₆ H ₅ CH TFMS H 6-OH CH ₃ p-AcO-C ₆ H ₅ C ₆ H ₅ CH TFMS H 6-OH CH ₃ p-NO ₂ -C ₆ H ₅ C ₆ H ₅ CH TFMS H 6-OH CH ₃ C ₆ H ₅ p- (CH ₃ ) ₂ NC ₆ H ₅ N TFMS H 6-OH CH ₃ C ₆ H ₅ -CH = CH- p- (CH ₃ ) ₂ NC ₆ H ₅ CH TFMS H 6-OH CH ₃ CH ₃ p- (CH ₃ ) ₂ NC ₆ H ₅ CH TFMS H 6-AcO CH ₃ C ₆ H ₅ C ₆ H ₅ CH TFMS H 6-AcO CH ₃ C ₆ H ₅ p- (CH ₃ ) ₂ NC ₆ H ₅ CH TFMS H 6-AcO CH ₃ C ₆ H ₅ p-NO ₂ -C ₆ H ₅ CH TFMS H 6-AcO CH ₃ C ₆ H ₅ p-OH-C ₆ H ₅ CH TFMS H 6-AcO CH ₃ C ₆ H ₅ p-OCH ₃ -C ₆ H ₅ CH TFMS H 6-AcO CH ₃ C ₆ H ₅ p-OAc-C ₆ H ₅ CH TFMS H 6-AcO CH ₃ C ₆ H ₅ p-NH ₂ -C ₆ H ₅ CH TFMS H 6-AcO CH ₃ C ₆ H ₅ p-NHCH ₃ -C ₆ H ₅ CH TFMS H 6-AcO CH ₃ C ₆ H ₅ p-NHAc-C ₆ H ₅ CH TFMS H 6-AcO CH ₃ C ₆ H ₅ -CH = CH- C ₆ H ₅ CH TFMS H 6-AcO CH ₃ C ₆ H ₅ -CH ₂ CH ₂ -C ₆ H ₅ CH TFMS H 6-AcO CH ₃ CH ₃ C ₆ H ₅ CH TFMS H 6- AcO CH ₃ HC ₆ H ₅ CH TFMS H 6-AcO CH ₃ p- (CH ₃ ) ₂ NC ₆ H ₅ C ₆ H ₅ CH TFMS H 6-AcO CH ₃ p-OH-C ₆ H ₅ C ₆ H ₅ CH TFMS H 6-AcO CH ₃ p-AcO -C ₆ H ₅ C ₆ H ₅ CH TFMS H 6-AcO CH ₃ p-NO ₂ -C ₆ H ₅ C ₆ H ₅ CH TFMS H 6-AcO CH ₃ C ₆ H ₅ p- (CH ₃ ) ₂ NC ₆ H ₅ N TFMS H 6-AcO CH ₃ C ₆ H ₅ -CH = CH- p- (CH ₃ ) ₂ NC ₆ H ₅ CH TFMS H 6-AcO CH ₃ CH ₃ p- (CH ₃ ) ₂ NC ₆ H ₅ CH TFMS ───────────────────────────────────

The compound of the present invention can be produced by various methods, and a typical production method thereof is shown in the following scheme as an example. However, the method for producing the compound of the present invention is not limited to these methods, and the scope of the compound of the present invention is not limited to those produced by this production method.

Embedded image

A compound represented by the formula (V) in the scheme (hereinafter referred to as "compound (V)"; the same applies hereinafter) and R ⁵ -C.
The desired compound (I) of the present invention can be obtained by condensing the aldehyde derivative represented by HO or the nitroso derivative represented by R ⁵ -NO under acidic conditions. This condensation reaction gives isomers of E-configuration or Z-configuration with respect to the exomethylene or imine moieties, but generally, when the reaction time of condensation is short, isomers of E-configuration and isomers of Z-configuration are obtained. When the reaction time of the condensation is long, the Z-configuration isomer is obtained as the main product when almost the same amount of the product is produced.

Compound (V) is a known compound (for example, J. C.
hem. Soc. Perkin Trans. 1.pp.2087-2097, 1993; J.
Chem. Soc. Perkin Trans. 1. pp.1871-1877, 1994; and Chem. Pharm. Bull., 40, pp.2267-2269, 1992), or according to a method commonly used in this field,
It is a compound that can be easily produced by those skilled in the art from readily available raw materials. For example, compound (V) is compound (I) obtained by oxidation of compound (II) or dehydration of compound (III).
It can be produced by subjecting V) to a quaternization reaction. Representative compounds of compounds (II) and (III) are described in publicly known publications (for compound (II) Chem.
Pharm. Bull., 38, pp. 1129-1135, 1990, etc.
II.) J. Chem. Soc. Perkin Trans. 1., pp.
2087-2097, 1993 etc.).

The reaction shown in the above scheme will be described in more detail. However, the production conditions of the compound (1) of the present invention are not limited to the specific details of these explanations, and those skilled in the art can understand these. It goes without saying that it is possible to make appropriate changes and modifications to the conditions.

Compound (IV) can be obtained by treating compound (II) with an oxidizing agent in the presence of a solvent. Examples of the solvent include halogenated hydrocarbon solvents such as chloroform, dichloromethane, carbon tetrachloride, etc .; ether solvents such as diethyl ether, tetrahydrofuran, dioxane; aromatic hydrocarbon solvents such as benzene, toluene, xylene; N, N An amide solvent such as dimethylformamide, acetamide, dimethylacetamide; an alcohol solvent such as methanol, ethanol, propanol; or a mixture thereof can be used. As oxidizing agents,
Examples thereof include 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) and chloranil. In the usual case, the reaction may be carried out in the temperature range of 0 to 200 ° C., preferably 10 to 100 ° C. for 5 minutes to 72 hours, preferably 20 minutes to 8 hours.

The corresponding compound (IV) can be obtained by treating the compound (III) with tosyl chloride or mesyl chloride in the presence of a base in a solvent. Compound (IV) can also be produced by treating compound (III) in a solvent in the presence of a dehydrating agent. Examples of the solvent include halogenated hydrocarbon solvents such as chloroform, dichloromethane, carbon tetrachloride, etc .; ether solvents such as diethyl ether, tetrahydrofuran, dioxane; aromatic hydrocarbon solvents such as benzene, toluene, xylene; N, N-dimethylformamide, acetamide,
An amide solvent such as dimethylacetamide; or a mixed solvent thereof can be used. As the base, triethylamine, sodium hydride, potassium t-butoxy, sodium carbonate or the like can be used, and as the dehydrating agent, p-toluenesulfonic acid, hydrochloric acid, sulfuric acid, hydrobromic acid, methanesulfonic acid, oxalic acid, etc. , Potassium hydrogen sulfate,
Alumina, n-butyllithium, phenyllithium, 1,8-
Diazabicyclo [5.4.0] undes-7-ene (DBU) and the like can be used. Usually, the reaction is -20 to 200 ℃,
It is preferably carried out in the temperature range of 0 to 100 ° C. for 5 minutes to 72 hours, preferably about 20 minutes to 8 hours.

Compound (V) can be obtained by treating compound (IV) with an alkylating reagent in the presence or absence of a solvent. Further, this reaction may be carried out by adding a base if desired. Examples of the solvent include halogenated hydrocarbon solvents such as chloroform, dichloromethane, carbon tetrachloride, etc .; ether solvents such as diethyl ether, tetrahydrofuran, dioxane; aromatic hydrocarbon solvents such as benzene, toluene, xylene; N, An amide-based solvent such as N-dimethylformamide, acetamide, dimethylacetamide; acetonitrile; or a mixed solvent thereof can be used. As the alkylating agent, trifluoromethanesulfonic acid methyl ester, methyl iodide,
Examples thereof include ethyl iodide, chloroacetonitrile, chloroacetamide, dimethyl sulfate and the like. When a base is used, sodium hydride, potassium t-butoxide, n-butyllithium, sodium carbonate, potassium carbonate or the like may be used. Usually, the reaction is -20 to 200 ℃,
It is preferably carried out in the range of 0 to 100 ° C. for 5 minutes to 72 hours, preferably about 20 minutes to 24 hours.

The compound (I) of the present invention can be obtained by treating the compound (V) with various aldehyde derivatives or various nitroso derivatives with an acidic solvent or with an acidic catalyst in the presence of a solvent. As an acidic solvent,
For example, acetic acid, formic acid and the like can be used, and as the acidic catalyst, acetic acid, formic acid, hydrochloric acid, sulfuric acid, p-toluenesulfonic acid, alumina and the like can be used. As the solvent when the acidic catalyst is used, other than acidic solvents such as acetic acid and formic acid, halogenated hydrocarbon solvents such as chloroform, dichloromethane, carbon tetrachloride and the like; ether solvents such as diethyl ether, tetrahydrofuran and dioxane; benzene, Aromatic hydrocarbon solvents such as toluene and xylene;
Amide solvents such as N, N-dimethylformamide, acetamide, dimethylacetamide; acetonitrile; or a mixed solvent thereof can be used. Usually, the reaction is carried out in the temperature range of 0 to 200 ° C, preferably 70 to 150 ° C.
It may be carried out for about minutes to 120 hours, preferably about 20 minutes to 72 hours.

The compound of the present invention has antitumor activity against various mammalian tumors and is useful as an active ingredient of antitumor agents. Without being bound to any particular theory, the compounds of the present invention have a strong inhibitory effect on both topoisomerase I and topoisomerase II present in mammals, and inhibit these DNA topoisomerases. Therefore, it is considered that the antitumor activity is selectively exerted on tumor cells. Therefore,
It is characterized in that it does not show cross resistance to tumor cells that have become resistant to existing antitumor agents, and has strong antitumor activity. The compounds of the present invention are also useful as inhibitors of DNA topoisomerase.

When the compound of the present invention is used as an antitumor agent, one or more compounds of the present invention may be administered orally or parenterally, but in general, the present invention is used. It is preferred to prepare a pharmaceutical composition containing one or more compounds of the above-mentioned as an active ingredient and use them orally or parenterally. Examples of the pharmaceutical composition include tablets, powders, granules, fine granules, capsules, solutions, syrups, elixirs, and preparations for oral administration such as oily or aqueous suspensions; or Examples thereof include injections for intravenous injection, intramuscular injection, or subcutaneous injection, infusions, transdermal or transmucosal absorption agents, and preparations for parenteral administration such as suppositories. Of these, aqueous injections and drops for intravenous administration, and preparations for oral administration are preferable. The form of the compound of the present invention, which is an active ingredient of a pharmaceutical composition, is not particularly limited, and hydrates, acid addition salts with pharmaceutically acceptable acids, and the like may be used. In particular, it is preferable to use a compound in the form of an acid addition salt for the production of an aqueous injection or drip.

For the production of a pharmaceutical composition, a pharmacologically and pharmaceutically acceptable pharmaceutical additive and a coating agent which are available to those skilled in the art, more specifically, an excipient, a disintegrant or Disintegration aids, binders, lubricants, coatings, dyes, diluents, bases, solubilizers or solubilizers, isotonic agents, pH adjusters, stabilizers, propellants, adhesives, etc. Can be used. For the production of a pharmaceutical composition suitable for oral administration, for example, an excipient such as glucose, lactose, D-mannitol, starch, or crystalline cellulose; a disintegrant or a disintegration aid such as carboxymethylcellulose, starch, or carboxymethylcellulose calcium. Agents; binders such as hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, or gelatin; lubricants such as magnesium stearate or talc;
A coating agent such as hydroxypropylmethyl cellulose, sucrose, polyethylene glycol or titanium oxide; a base such as petrolatum, liquid paraffin, polyethylene glycol, gelatin, kaolin, glycerin, purified water, or hard fat can be used.

For producing a pharmaceutical composition suitable for transdermal or transmucosal administration, propellants such as CFC, diethyl ether, or compressed gas; sodium polyacrylate, polyvinyl alcohol, methyl cellulose, polyisobutylene, polybutene, etc. Adhesives such as cotton cloth or base cloth such as plastic sheet can be used, and for preparations suitable for injection or infusion, aqueous solutions such as distilled water for injection, physiological saline, propylene glycol, etc. Agents or solubilizers that can constitute agents; isotonic agents such as glucose, sodium chloride, D-mannitol, and glycerin; pharmaceutical additives such as pH adjusting agents such as inorganic acids, organic acids, inorganic bases or organic bases A thing can be used. However, the additive for formulation and the coating agent used for producing the pharmaceutical composition are not limited to these, and should be appropriately selected depending on the type of the pharmaceutical composition to be produced and the therapeutic purpose.

The type of tumor to be treated by the pharmaceutical composition of the present invention is not particularly limited, and includes brain tumor, esophageal cancer, tongue cancer, gastric cancer, lung cancer, kidney cancer, liver cancer, pancreatic cancer, duodenum or small intestine cancer, A solid cancer such as colorectal cancer, skin cancer, uterine cancer, breast cancer, or prostate cancer, or a non-solid cancer such as leukemia or acute lymphoma can be treated. The dose of the above-mentioned pharmaceutical composition is in the range of 1 mg to 2 g, preferably about 5 mg to 1 g per day for an adult, as the weight of the compound of the present invention as an active ingredient. It should be adjusted according to the type of tumor to be treated, the age and condition of the patient, etc. Hereinafter, the present invention will be described more specifically with reference to Examples, but the scope of the present invention is not limited to these Examples.

[0038]

【Example】

Example 1: 1-methyl-4- (Z)-(p-dimethylaminophenylmethyi
Len) -2-phenyl-4H-pyrazolo [1,5-a] indolium ・
Trifluoromethanesulfonate 1-methyl-2-phenyl-4H-pyrazolo [1,5-a] indolium trifluoromethanesulfonate (hereinafter, this compound is referred to as “compound (1)” in Examples) ( 0.30
g, 0.82 mmol) and p-dimethylaminobenzaldehyde (0.14 g, 0.94 mmol) were added to glacial acetic acid (30 ml), and the mixture was heated under reflux for 20 hours under a nitrogen stream. The reaction mixture was concentrated and recrystallized from ethanol to give the title compound (0.32g, 86%). Melting point: 237.0-237.5 ° C. IR (KBr) cm ^-1 : 1583 (s), 1529, 1464, 1360, 1325, 1
188, 1161, 1095 (s). ¹ H-NMR (DMSO-d ₆ ) δ: 3.11 (6H, s), 4.43 (3H, s), 6.9
3 (2H, d, J = 9.0Hz), 7.45 (1H, s), 7.56 (2H, m), 7.70
(3H, m), 7.82 (2H, m), 7.84 (2H, d, J = 9.0Hz), 8.14 (1
H, m), 8.22 (1H, m), 8.30 (1H, s). Elemental analysis calculated for C ₂₇ H ₂₄ F ₃ N ₃ O ₃ S (%) C, 61.47;
H, 4.59; N, 7.97. Actual value (%) C, 61.44; H, 4.66; N,
7.98. In the above reaction, under the condition of heating and stirring for 1.5 hours, E-
The isomer was obtained in almost the same amount as the Z-isomer. ¹ H-NMR (DMSO-d ₆ ) δ: 3.10 (s), 4.43 (s), 6.52 (d, J = 9H
z), 7.49 (s), 7.54-7.58 (m), 7.68-7.19 (m), 7.81 (d, J
= 9Hz), 7.81-7.82 (m), and 8.28 (s).

Example 2: 1-Methyl-2-phenyl-4- (Z) -phen
Nilmethylene-4H-pyrazolo [1,5-a] indolium tri
Fluoromethanesulfonate compound (1) (0.50 g, 1.26 mmol) and benzaldehyde
(0.17 ml, 1.67 mmol) was added to acetic acid (50 ml), and the mixture was heated with stirring for 20 hours. The reaction solution was concentrated, and the obtained residue was purified by silica gel column chromatography (dichloromethane-
Methanol = 9: 1) to give the title compound (0.61 g, 99%)
. Melting point: 233-235 ° C. IR (KBr) cm ^-1 : 1627, 1544, 1467, 1383, 1263, 122
6, 1157, 1034, 766, 700, 641. ¹ H-NMR (DMSO-d ₆ ) δ: 4.46 (3H, s), 7.39 (1H, s), 7.59
-7.81 (H, m), 7.90 (2H, d, J = 7Hz), 8.18 (1H, d, J = 8H
z), 8.18 (1H, d, J = 8Hz), 8.34 (1H, d, J = 7Hz), 8.55 (1
H, s). Elemental analysis calculated for C ₂₅ H ₁₉ F ₃ N ₂ O ₃ S (%) C, 62.07;
H, 3.95; N, 5.78. Actual value (%) C, 62.07; H, 4.01; N,
5.82.

Example 3: 1-Methyl-4- (Z)-(p-nitrophenyl
Methylene) -2-phenyl-4H-pyrazolo [1,5-a] indriu
Trifluoromethanesulfonate compound (1) (0.50 g, 1.37 mmol) and p-nitrobenzaldehyde (0.25 g, 1.26 mmol) were added to glacial acetic acid (50 ml).
The mixture was heated under reflux for 10 hours. The reaction solution was concentrated and recrystallized from methanol-isopropyl ether to obtain the title compound (0.53
g, 79%). Melting point: 188-190 ° C. IR (KBr) cm ^-1 : 1518, 1338, 1262 (s), 1153, 1028. ¹ H-NMR (DMSO-d ₆ ) δ: 4.48 (1H, s), 7.45 (1H , s), 7.6
1-7.81 (7H, m), 8.09 (2H, d, J = 8.8Hz), 8.19 (1H, d, J
= 7.8Hz), 8.33 (1H, d, J = 7.1Hz), 8.45 (2H, d, J = 8.8H
z), 8.61 (1H, s). Elemental analysis calculated for C ₂₅ H ₁₈ F ₃ N ₃ O ₅ S (%) C, 56.71;
H, 3.43; N, 7.94. Actual value (%) C, 56.57; H, 3.47; N,
7.88.

Example 4: 1-Methyl-4- (Z)-(p-hydroxyphene
Nylmethylene) -2-phenyl-4H-pyrazolo [1,5-a] India
The lithium trifluoromethanesulfonate compound (1) (100 mg, 0.25 mmol) and p-hydroxybenzaldehyde (37 mg, 0.30 mmol) were added to acetic acid (10 ml), and the mixture was heated with stirring under a nitrogen stream for 22.5 hours. The reaction solution was concentrated, and the obtained residue was purified by silica gel column chromatography (dichloromethane-methanol = 19: 1) to obtain the title compound (87 mg, 69%). Melting point: 113-115 ° C. IR (KBr) cm ^-1 : 3242, 1599, 1580, 1465, 1280, 126
8, 1243, 1224. 1162, 1029. ¹ H-NMR (DMSO-d ₆ ) δ: 4.45 (3H, s), 7.03 (2H, d, J = 8.
5Hz), 7.52 (1H, s), 7.52-7.72 (7H, m), 7.83 (2H, d, J
= 8.5Hz), 8.16 (1H, d, J = 8.5Hz), 8.26-8.30 (1H, m), 8.
40 (1H, s).

Example 5: 1-Methyl-4- (Z)-(p-acetoxyphe
Nylmethylene) -2-phenyl-4H-pyrazolo [1,5-a] India
The lithium trifluoromethanesulfonate compound (1) (100 mg, 0.25 mmol) and p-acetoxybenzaldehyde (0.05 ml, 0.35 mmol) were added to acetic acid (10 ml), and the mixture was heated and stirred for 14 hours under a nitrogen stream. The reaction solution was concentrated, and the obtained residue was purified by silica gel column chromatography (dichloromethane-methanol = 9: 1) to obtain the title compound (110 mg, 80%). Melting point: 197-198 ° C. IR (KBr) cm ^-1 : 1764, 1467, 1276, 1264, 1229, 119
9, 1162, 1031, 764, 638. 1 H-NMR (DMSO-d 6) δ: 2.23 (3
H, s), 4.46 (3H, s), 7.42 (2H, d, J = 8.6Hz), 7.43 (1H,
s), 7.59-7.82 (7H, m), 7.96 (2H, d, J = 8.6Hz), 8.18
(1H, d, J = 7.3Hz), 8.32 (1H, d, J = 7.6Hz), 8.52 (1H,
s). Elemental analysis C ₂₇ H ₂₁ F ₃ N ₂ O ₅ S ・ Calculated value (%) for 1 / 2H ₂ O
C, 58.79; H, 4.02; N, 5.08. Actual value (%) C, 59.03; H,
3.79; N, 5.05.

The compound of Example 5 was also obtained by the following method. The compound 1-methyl-4- (Z)-(p-hydroxyphenylmethylene) -2-phenyl-4H-pyrazolo [1,5-a] indolium (250 mg, 0.50 mmol) obtained in Example 4 was replaced with pyridine (10 ml) and acetic anhydride (0.47 ml), and the mixture was stirred at room temperature for 5 hours. The reaction solution was concentrated, water was added, and the mixture was extracted with dichloromethane,
The extract was washed with 1N-hydrochloric acid and saturated brine, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (dichloromethane-methanol = 9: 1).
This gave the title compound (137 mg, 51%).

Example 6: 1-Methyl-4- (Z)-(p-dimethylamino
Phenylmethylene) -2-styryl-4H-pyrazolo [1,5-a] i
Indolium trifluoromethanesulfonate 2- (E) -styryl- 3-hydroxy-3a, 4-dihydro-3H-pyrazolo [1,5-a] indole (0.63 g, 2.1 mmol) in anhydrous dichloromethane (38 ml) The mixture was dissolved in water, methanesulfonyl chloride (0.25 ml, 3.1 mmol) and triethylamine (0.60 ml, 4.3 mmol) were added under ice cooling, and the mixture was stirred for 4 hours. After diluting the reaction solution with water and adjusting to alkaline with sodium carbonate,
The organic layer was extracted, washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated. The residue was purified by silica gel column chromatography (hexane-ethyl acetate = 5:
1) to give 2-styryl-4H-pyrazolo [1,5-a] indole (0.48 g, 91%). Melting point: 148-149 ° C. IR (KBr) cm ^-1 : 2908, 1619, 1591, 1480, 1401, 1306
^{. 1 H-NMR (DMSO-} d 6) δ: 3.86 (2H, s), 6.50 (1H, s), 7.
11-7.55 (10H, m), 7.62 (1H, br d, J = 8.0Hz).

The compound obtained in the above reaction (1.0 g, 3.87 m
Anhydrous dichloromethane (20 ml) trifluoromethanesulfonic acid methyl ester (0.88 ml, 7.74 mmol) was added to (mol) and the mixture was stirred at room temperature for 48 hours under a nitrogen stream. The reaction solution was concentrated and recrystallized from methanol-ethyl acetate to give 1-methyl-2-
Styryl-4H-pyrazolo [1,5-a] indolium trifluoromethanesulfonate was obtained (1.35 g, 82%). Melting point: 223.5-225.0 ° C. IR (KBr) cm ^-1 : 1539, 1468, 1291, 1247, 1224, 115
4, 1027. ¹ H-NMR (DMSO-d ₆ ) δ: 4.38 (2H, s), 4.49 (3H, s), 7.
43-7.86 (11H, m), 8.10 (1H, br.d, J = 7.81Hz). Elemental analysis C ₂₂ H ₂₂ F ₃ N ₃ O ₃ S Calculated value (%) C, 56.86;
H, 4.06; N, 6.63. Actual value (%) C, 56.98; H, 3.97; N,
6.63.

The compound obtained in the above reaction (500 mg, 1.18
mmol) and p-dimethylaminobenzaldehyde (229 mg,
1.53 mmol) was added to glacial acetic acid (50 ml), and the mixture was heated under reflux for 12 hours. The reaction solution was concentrated and recrystallized from ethanol to obtain the title compound (336 mg, 51%). Melting point: 144.5-146.5 ° C (decomposition). IR (KBr) cm ^-1 : 1579, 1529, 1369, 1261, 1159, 103
^{1, 638. 1 H-NMR (} DMSO-d 6) δ: 3.13 (6H, s), 4.59 (3H, s), 6.9
6 (2H, d, J = 8.79Hz), 7.45-7.99 (12H, m), 8.07-8.12 (1
H, m), 8.15-8.20 (1H, m), 8.24 (1H, s). Elemental analysis calculated for C ₂₉ H ₂₆ F ₃ N ₃ O ₃ S (%) C, 62.92;
H, 4.73; N, 7.59. Actual value (%) C, 62.46; H, 4.65; N,
7.62.

Example 7: 1,2-dimethyl-4- (Z)-(p-dimethyla
Minophenylmethylene) -4H-pyrazolo [1,5-a] indori
Umium trifluoromethanesulfonate 2-methyl-2,3,3a, 4-tetrahydro-1H-pyrazolo [1,5-a]
Indole (2.23 g, 12.8 mmol) was added to anhydrous dichloromethane
Dissolve in (80 ml) 2,3-dichloro-5,6-dicyano-1,4-
Benzoquinone (6.39 g, 28.2 mmol) was added, and the mixture was stirred at room temperature for 2 hr. The reaction mixture was diluted with ether (30 ml), the insoluble material was filtered off, the filtrate was washed with 10% sodium hydroxide solution and saturated brine, dried over anhydrous sodium sulfate, and concentrated.
The residue was recrystallized from ether-n-hexane to give 2-methyl-4H-
Pyrazolo [1,5-a] indole was obtained (1.0 g, 46%). Melting point: 66.0-67.5 ° C. IR (KBr) cm ^-1 : 1619, 1592, 1565, 1545, 1472, 140
2, 1332, 1301, 1199, 1129, 1097, 1003. ¹ H-NMR (DMSO-d ₆ ) δ: 2.42 (3H, s), 3.81 (2H, s), 6.0
6 (1H, s), 7.09-7.17 (1H, m), 7.32-7.43 (2H, m), 7.56
(1H, d).

Compound obtained in the above reaction (0.5 g, 2.9 mm
ol) was dissolved in anhydrous dichloromethane (20 ml) and trifluoromethanesulfonic acid methyl ester (2.66 ml, 23.5 m
(mol) was added in two portions. After 42 hours, the precipitate was collected by filtration and recrystallized from methanol to give 1,2-dimethyl-4H-pyrazolo [1,5-
a] Indolium trifluoromethanesulfonate was obtained (0.45 g, 46%). Melting point: 210-212 ° C. IR (KBr) cm ^-1 : 1545, 1485, 1398, 1267, 1227, 115
7, 1029. ¹ H-NMR (DMSO-d ₆ ) δ: 2.58 (3H, s), 4.31 (2H, s), 4.3
7 (3H, s), 6.91 (1H, s), 7.48 (1H, t), 7.59 (1H, t), 7.
75 (1H, d), 8.06 (1H, d). Elemental analysis calculated for C ₁₃ H ₁₃ F ₃ N ₂ O ₃ S (%) C, 46.71;
H, 3.92; N, 8.38. Actual value (%) C, 46.41; H, 3.91; N,
8.21.

Compound obtained in the above reaction (500 mg, 1.50
mmol) and p-dimethylaminobenzaldehyde (291 mg,
1.95 mmol) was added to acetic acid (50 ml), and the mixture was heated with stirring for 32 hours. The reaction solution was concentrated and recrystallized from methanol to obtain the title compound (576 mg, 85%). Melting point: 249-251 ° C. IR (KBr) cm ^-1 : 1585, 1532, 1378, 1362, 1266, 119
^{2, 1164, 1142, 1031. 1} H-NMR (DMSO-d 6) δ: 2.58 (3H, s), 3.02 (6H, s), 4.3
6 (3H, s), 6.88 (2H, ABtype, J = 8.78Hz), 7.20 (1H, s),
7.44-7.54 (2H, m), 7.72 (2H, AB type, J = 8.8Hz), 8.0
0-8.04 (1H, m), 8.01-8.15 (1H, m), 8.17 (1H, s). Elemental analysis C ₂₂ H ₂₂ F ₃ N ₃ O ₃ S calculated value (%) C, 56.76;
H, 4.77; N, 9.03. Actual value (%) C, 56.57; H, 4.64; N,
9.03.

Example 8: 1-Methyl-4- (Z)-(p-dimethylamino)
Phenylimino) -2-phenyl-4H-pyrazolo [1,5-a] in
Dorium / trifluoromethanesulfonate compound (1) (0.1 g, 0.27 mmol) and N, N-dimethyl-p-nitrosoaniline (0.05 ml, 0.25 mmol) were added to acetic acid (10 ml) under a nitrogen stream at 1.5. The mixture was heated and stirred for an hour. The reaction mixture was concentrated, and the obtained residue was purified by silica gel column chromatography (dichloromethane-methanol = 95: 5) to give the title compound (0.12 g, 90%). Melting point: 204-206 ° C. IR (KBr) cm ^-1 : 1618, 1551, 1364, 1261 (s), 1151
^{(s), 1030. 1 H-} NMR (DMSO-d 6) δ: 3.08 (6H, s), 4.45 (3H, s), 6.9
0 (2H, d, J = 9Hz), 7.23 (1H, s), 7.45 (2H, d, J = 9.0H
z), 7.61 (1H, t, J = 7.6Hz), 7.65-7.77 (8H, m), 8.08 (1
H, d, J = 7.4Hz), 8.15 (1H, d, J = 8.0Hz). Elemental analysis C ₂₆ H ₂₃ F ₃ N ₄ O ₃ S Calculated value (%) C, 59.08;
H, 4.39; N, 10.60. Actual value (%) C, 60.02; H, 4.27; N,
10.10.

Example 9: Antitumor effect of the compound of the present invention Two types of tumor cells, P388 murine leukemia cells and PC-6 (lung cancer) cells, P388 was 5.0 × 10 ² cells / 150 μl / well, P388
C-6 was seeded on a 96-well microplate at 5.0 × 10 ³ cells / μl / well, P388 was for 2 hours, and PC-6 was
After 24 hours, 50 μl / well of the sample was added. Then, the cells were cultured for 3 days, and MTT [3- (4,5-dimethylthiazol-2-yl)-
2,5-diphenyl-2H-tetrazolium bromide] 5 mg /
20 μl / well of ml solution was added. After 4 hours, remove the culture solution and add dimethyl sulfoxide 150 μl / well to obtain 540n.
Absorbance was measured at m. The antitumor effect was determined by setting the GI ₅₀ value (μg / μg
ml).

Example 10: DNA topoisomers of compounds of the invention
Inhibitory action 25 mM Tris-HCl (pH 7.5), 50 mM KCl, 5 mM MgCl ₂ , 0.25
mM EDTA 2Na salt, 0.25 mM DTT (dithiothreitol), 15
DNA topoisomerase (0.5 μg SV was added to the reaction buffer consisting of μg / ml bovine serum albumin and 5% glycerol).
40 Minimal amount of topoisomerase I that completely relaxes DNA
(Topo I) or topoisomerase II (Topo II) and
0.5 μl of test substance or solvent (DMSO) was added. Topo
For II, an additional 10 mM ATP was added. 10 at 37 ° C
After allowing to stand for 0.5 minutes, 0.5 μg of SV40 DNA was added, and the mixture was reacted at the same temperature for 10 minutes. Stop solution (1% SDS, 20 mM EDTA 2Na
Salt, 0.5 mg / ml proteinase K) 5 μl, and
The reaction was stopped by treating at 7 ° C for 30 minutes.

10 mM Na ₂ HPO ₄ , 31.3% sucrose and 0.3%
After mixing with 5 μl of buffer containing bromophenol blue, 2 μg / ml chloroquine, 10 mM EDTA, 30 mM NaH ₂ PO ₄
Electrophoresis (50 mA, 20V, overnight) on 0.8% agarose gel in the presence of 36 mM Tris-HCl (pH 7.8), and then the relaxed supercoil (form Ir) DNA and supercoiled (form I) DNA And open circular (form II) DNA were separated. After electrophoresis, the gel was stained with 0.05% ethidium bromide and exposed to UV light (302
(nm) and the photograph was recorded. The amount of DNA was quantified by a densitometer. The inhibition rate of Topo I or Topo II was calculated according to the following formula. In the formula, Flrt and Flrc are the ratio of Form Ir DNA to the total DNA treated with Topo I and Topo II in the presence and absence of the test substance, respectively. Also, Flr is Topo I and Topo II untreated.
It is the ratio of Form Ir DNA mixed in DNA. The concentration (IC ₅₀ ) of each test substance giving 50% inhibition was determined from the dose-response curve. Inhibition rate = [1- (Flrt-Flr) / (Flrc-Flr)] x 100

The results obtained from the above tests are shown in Table 2 (the numbers in the table are IC ₅₀ values: μg / ml). As a control,
The results of the same test using 7-ethyl-10-hydroxycamptothecin (control drug A) having an inhibitory activity on topoisomerase I and etoposide (control drug B) inhibiting topoisomerase II are also shown. From these results, it is clear that the compound of the present invention has a strong inhibitory activity against both topoisomerase I and topoisomerase II and exhibits a strong cytostatic effect on tumor cells.

[0055]

[Table 2] ────────────────────────── Test compound P388 PC-6 TopoI inhibition TopoII inhibition ───────── ───────────────── Example 1 0.0086 0.037 0.54 0.38 Example 2 0.197 0.211 ＞ 10 3.98 Example 3 0.668 0.828 4.76 3.50 Example 4 1.350 1.350 0.58 0.63 Example 5 0.928 0.492 4.94 0.84 Example 6 0.011 0.021 5.52 0.54 Example 7 0.010 0.035 1.46 4.74 Example 8 0.293 0.164 0.50 0.39 Control agent A 0.0031 0.0007 8.9 NT Control agent B 0.015 0.144 NT 6.99 ───────────────────── ────── NT Not tested

[0056]

The compound of the present invention potently inhibits topoisomerase I and topoisomerase II and has a growth inhibitory action on tumor cells. Therefore, it is extremely useful as an active ingredient of an antitumor agent.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI technical display area C07D 209: 00 231: 00)

Claims (24)

[Claims] 1. The following general formula (I): [In the formula, Q represents CH or a nitrogen atom; R ¹ and R ² are each independently a hydrogen atom, a halogen atom, a C _1-6 alkyl group,
Amino group, C _1-6 alkylamino group, C _1-6 alkylcarbonylamino group, hydroxy group, C _1-6 alkoxy group, thiol group, C _1-6 alkylthio group, cyano group, carbamoyl group, nitro group, acyl A group or an acyloxy group;
R ³ is a C _1-6 alkyl group (C _1-6 alkyl group is a halogen atom, amino group, C _1-6 alkylamino group, C _1-6 alkylcarbonylamino group, hydroxy group, C _1-6 alkoxy group ,
A thiol group, a C _1-6 alkylthio group, a cyano group, a carbamoyl group, a nitro group, an acyl group and an acyloxy group, which may have one or more substituents); R ⁴ is a hydrogen atom, a C _1-6 alkyl group, a phenyl group, a phenyl-substituted C _1-6 alkyl group, or a phenyl-substituted C _2-6 alkenyl group (C _1-6 alkyl group, phenyl group,
The phenyl group portion of the phenyl-substituted C _1-6 alkyl group, or the phenyl group portion of the phenyl-substituted C _2-6 alkenyl group is a halogen atom, a C _1-6 alkyl group, an amino group, a C _1-6 alkylamino group, C _1-6 alkylcarbonylamino group, hydroxy group, C _1-6 alkoxy group, thiol group, C _1-6 alkylthio group, cyano group, carbamoyl group, nitro group, acyl group,
And a substituent selected from the group consisting of acyloxy groups
Or may have two or more); R ⁵ is a C _1-6 alkyl group, a phenyl group, a phenyl-substituted C _1-6 alkyl group or a phenyl-substituted C _2-6 alkenyl group (C _1-6 The alkyl group, the phenyl group, the phenyl group portion of the phenyl-substituted C _1-6 alkyl group, or the phenyl group portion of the phenyl-substituted C _2-6 alkenyl group is a halogen atom, a C _1-6 alkyl group, an amino group, C _{1- 6}
Alkylamino group, C _1-6 alkylcarbonylamino group,
Hydroxy group, C _1-6 alkoxy group, thiol group, C _1-6 alkylthio group, cyano group, carbamoyl group, nitro group,
1 or 2 or more substituents selected from the group consisting of an acyl group and an acyloxy group are shown);
And X ^- represents a salt-forming anion (wherein, X is when any one of the substituents of from R ¹ R ⁵ is a proton-donating group ^- may not be present) represented by] Compound. 2. The compound according to claim 1, wherein R ³ is a C _1-6 alkyl group. 3. The compound according to claim 1, wherein R ³ is a methyl group. 4. The compound according to claim 1, wherein R ¹ and R ² are simultaneously hydrogen atoms. 5. R ¹ is a hydrogen atom, R ² is substituted at the 6-position, and R ² is an amino group, a C _1-6 alkylamino group, a hydroxy group, a halogen atom, or an acyloxy group. Item 4. The compound according to any one of items 1 to 3. 6. The compound according to claim 5, wherein R ² is an amino group, a hydroxy group, a chlorine atom, a fluorine atom or an acetoxy group. 7. R ⁴ is a hydrogen atom, a C _1-6 alkyl group, a phenyl group, a phenyl-substituted C _1-6 alkyl group, or a phenyl-substituted group.
It is a C _2-6 alkenyl group, or one substituent selected from the group consisting of a C _1-6 alkylamino group, a hydroxy group, an acyloxy group, and a nitro group is a phenyl group substituted at the p-position. A compound according to any one of claims 1 to 6. 8. The compound according to claim 7, wherein R ⁴ is a C _1-6 alkyl group, a phenyl group, a phenyl-substituted C _1-6 alkyl group, or a phenyl-substituted C _2-6 alkenyl group. 9. The compound according to claim 7, wherein R ⁴ is a methyl group, a phenyl group, a phenethyl group, or a 2-phenylethenyl group. 10. The compound according to any one of claims 1 to 9, wherein R ⁵ is a phenyl group or a p-substituted phenyl group. 11. The p-position substituent is an amino group, a C _1-6 alkylamino group, a hydroxy group, an acyloxy group, a nitro group,
_11. The compound according to claim 10, which is a C _1-6 alkoxy group or a C _1-6 alkylcarbonylamino group. 12. The method according to claim 10, wherein R ⁵ is a phenyl group, a p-diC _1-4 alkylaminophenyl group, a p-nitrophenyl group, a p-hydroxyphenyl group or a pC _1-4 alkylcarbonyloxyphenyl group. The described compound. 13. The compound according to claim 10, wherein R ⁵ is a phenyl group, a p-dimethylaminophenyl group, a p-nitrophenyl group, a p-hydroxyphenyl group or a p-acetoxyphenyl group. 14. The compound according to any one of claims 1 to 13, wherein Q is CH. 15. The compound according to claim 1, wherein X ⁻ is a trifluoromethanesulfonic acid anion. 16. The following compound: 1-methyl-4- (Z)-(p-dimethylaminophenylmethylene) -2-phenyl-4H-pyrazolo [1,5-a] indolium trifluoromethanesulfonate. 1-Methyl-2-phenyl-4- (Z) -phenylmethylene
-4H-Pyrazolo [1,5-a] indolium trifluoromethanesulfonate; 1-methyl-4- (Z)-(p-nitrophenylmethylene) -2-phenyl-4H-pyrazolo [1,5- a] Indolium trifluoromethanesulfonate; 1-methyl-4- (Z)-
(p-Hydroxyphenylmethylene) -2-phenyl-4H-pyrazolo [1,5-a] indolium trifluoromethanesulfonate; 1-methyl-4- (Z)-(p-acetoxyphenylmethylene) -2 -Phenyl-4H-pyrazolo [1,5-a] indolium trifluoromethanesulfonate; 1-methyl-4- (Z)-(p-dimethylaminophenylmethylene) -2-styryl-4H-pyrazolo [1 , 5-a] Indolium trifluoromethanesulfonate; 1,2-dimethyl-4- (Z)-(p-dimethylaminophenylmethylene) -4H-pyrazolo [1,5-a] indolium trifluoromethane Sulfonate; and 1-methyl-4- (Z)-(p-
Dimethylaminophenylimino) -2-phenyl-4H-pyrazolo [1,5-a] A compound selected from the group consisting of indolium trifluoromethanesulfonate. 17. A DNA topoisomerase inhibitor comprising the compound according to any one of claims 1 to 16. 18. A DNA topoisomerase inhibitor of eukaryotic cells, which comprises the compound according to any one of claims 1 to 16. 19. The inhibitor according to claim 17 or 18, which has an inhibitory effect on topoisomerase I and topoisomerase II. 20. A medicine comprising the compound according to any one of claims 1 to 16. 21. A medicine comprising the compound according to any one of claims 1 to 16 as an active ingredient. 22. The use as an antitumor agent according to claim 20 or 21.
The medicament according to item 1. 23. A pharmaceutical composition comprising the compound according to any one of claims 1 to 16 as an active ingredient. 24. The pharmaceutical composition according to claim 23, which is used as an antitumor agent.