JPWO2019004413A1 - Indoline derivative and cytotoxic agent containing the same - Google Patents

Abstract

The present invention provides 1- (chloromethyl) -5-hydroxy-1,2-dihydro-3H corresponding to the 6-position in the 1,2,9,9a-tetrahydrocyclopropa [c] benz [e] indol-4-one skeleton. An object of the present invention is to provide a novel Duocarmycin derivative having a functional group at the 7-position of the -benzo [e] indole skeleton and exhibiting high cytotoxicity. The present invention provides an indoline derivative represented by the following formula or a pharmaceutically acceptable salt thereof.

Description

  The present invention relates to indoline derivatives and cytotoxic agents containing the same.

  Duocarmycin and its derivatives are one of the natural products having extremely strong cytotoxicity due to alkylation of DNA, and many derivatives have been reported so far (Non-Patent Document 1).

  Among the reported derivatives, 1,2,9,9a-tetrahydrocyclopropa [c] benz [e] indol-4-one skeleton or 1- (chloromethyl) -5-hydroxy-1,2-dihydro that serves as a precursor thereof. It is known that a compound having a -3H-benzo [e] indol skeleton is more easily synthesized than a skeleton of a natural product and has excellent chemical stability while maintaining strong cytotoxicity. (Non-patent document 1).

  The Duocarmycin derivative has a structure for alkylating DNA and a structure for promoting alkylation by interacting with DNA. For example, 1,2,9,9a-tetrahydrocyclopropa [c] benz [ The e] indol-4-one skeleton is a structure that alkylates DNA via a cyclopropane structure.

  Up to now, as the substituents on the 1,2,9,9a-tetrahydrocyclopropa [c] benz [e] indol-4-one skeleton, there have been reports centering on the substituents at the 3-position, 5-position and 7-position. It is known that the substituent at the 7-position has a great influence on cytotoxicity. The reason is that the structure of the substituent at the 7-position allows the cyclopropane structure to be close to the DNA and take a position suitable for alkylation of the DNA. In that case, the substituent at the 7-position is nitrile. It is known that a group and a methoxy group are preferable (Non-Patent Document 1). On the other hand, there has been no report of a derivative having a substituent at the 6-position of 1,2,9,9a-tetrahydrocyclopropa [c] benz [e] indol-4-one skeleton, which is a precursor thereof and has the above skeleton 1- (chloromethyl) -5-hydroxy-1,2-dihydro-3H-benzo [e] indol skeleton corresponding to the 6-position of trimethylsilylethyl ester group at the 7-position has been reported (non-patent document) Reference 2).

Ghosh et al., Current Topics in Medicinal Chemistry, 2009, Volume 9, p. 1494-1524 Tietze et al., International Journal of Molecular Science, 2008, vol. 9, p. 821-837

  However, as described above, 1- (chloromethyl) -5-hydroxy-1,2-dihydrocorresponds to the 6-position in the 1,2,9,9a-tetrahydrocyclopropa [c] benz [e] indol-4-one skeleton. A Duocarmycin derivative having a substituent other than a trimethylsilylethyl ester group at the 7-position of the -3H-benzo [e] indol skeleton and exhibiting high cytotoxicity has not been reported so far.

  A novel Duocarmycin derivative having a substituent other than trimethylsilylethyl ester group at the 7-position of 1- (chloromethyl) -5-hydroxy-1,2-dihydro-3H-benzo [e] indol skeleton and exhibiting high cytotoxicity is , 1,2,9,9a-tetrahydrocyclopropa [c] benz [e] indol-4-one, which is useful for verifying the effect of the substituent at the 6-position on the cytotoxicity on cytotoxicity, and is a new cytotoxic agent Will lead to the creation of.

  Therefore, the present invention relates to 1- (chloromethyl) -5-hydroxy-1,2-dihydro-corresponding to the 6-position in the 1,2,9,9a-tetrahydrocyclopropa [c] benz [e] indol-4-one skeleton. It is an object to provide a novel Duocarmycin derivative having a functional group at the 7-position of the 3H-benzo [e] indol skeleton and exhibiting high cytotoxicity.

  The present inventors have conducted extensive studies to achieve the above object, and as a result, found that a novel indoline derivative having a specific functional group or a pharmaceutically acceptable salt thereof exhibits high cytotoxicity, and the present invention Has been completed.

［式中、Ｘは、−ＣＯ_２Ｍｅ、−ＣＯ_２Ｅｔ、−Ｃ（＝Ｏ）ＮＨ（ＣＨ_２）_２Ｙ又は−ＮＨＣ（＝Ｏ）Ｒ^２を表し、Ｙは、一個の水素原子がＮＨ_２で置換されたフェニル基を表し、Ｒ^１は、水素原子又はメトキシ基を表し、Ｒ^２は、メチル基、エチル基又は−ＣＨ_２ＮＨ_２を表す。］That is, the present invention provides an indoline derivative represented by the following general formula (I) or a pharmaceutically acceptable salt thereof.

[In the _{formula, X, -CO 2 Me, -CO 2 Et} , represent _{-C (= O) NH (CH} 2) 2 Y or ^{-NHC (= O) R 2,} Y is the one hydrogen atom It represents a phenyl group substituted with NH ₂ , R ¹ represents a hydrogen atom or a methoxy group, and R ² represents a methyl group, an ethyl group or —CH ₂ NH ₂ . ]

In the indoline derivative represented by the general formula (I) or the pharmaceutically acceptable salt thereof, R ¹ is preferably a hydrogen atom.

In the indoline derivative represented by the general formula (I) or a pharmaceutically acceptable salt thereof, R ² is preferably a methyl group or —CH ₂ NH ₂ , and is —CH ₂ NH _2. Is more preferable.

In Salts indoline derivative or a pharmacologically acceptable represented by the above general formula (I), X is a _{-C (= O) NH (CH} 2) 2 Y or -NHC (= O) ^{R 2} It is preferable that it is present, and —C (═O) NH (CH ₂ ) ₂ Y or —NHC (═O) CH ₂ NH ₂ is more preferred.

  In the indoline derivative represented by the general formula (I) or the pharmaceutically acceptable salt thereof, Y is preferably a 4-aminophenyl group.

In Salts indoline derivative or a pharmacologically acceptable represented by the above general formula (I), X is a _{-C (= O) NH (CH} 2) 2 Y or -NHC (= O) ^{R 2} And R ² is preferably a methyl group or —CH ₂ NH ₂ .

  In this case, high cytotoxicity can be expected.

In Salts indoline derivative or a pharmacologically acceptable represented by the above general formula (I), X is a _{-C (= O) NH (CH} 2) 2 Y or -NHC (= O) ^{R 2} And Y is a phenyl group in which one hydrogen atom is replaced with NH ₂ , R ¹ is a hydrogen atom or a methoxy group, and R ² is preferably a methyl group or —CH ₂ NH _2. ..

  In this case, high cytotoxicity can be expected.

In the indoline derivative represented by the general formula (I) or a pharmaceutically acceptable salt thereof, X is —CO ₂ Me, —CO ₂ Et, —C (═O) NH (CH ₂ ) ₂ Y. Or —NHC (═O) R ² , Y is a phenyl group in which one hydrogen atom is replaced with NH ₂ , R ¹ is a hydrogen atom, R ² is a methyl group, an ethyl group or it is preferred is -CH ₂ NH _2.

  In this case, high cytotoxicity can be expected.

In Salts indoline derivative or a pharmacologically acceptable represented by the above general formula (I), X is a _{-C (= O) NH (CH} 2) 2 Y or -NHC (= O) ^{R 2} And R ¹ is a hydrogen atom, and R ² is preferably a methyl group or —CH ₂ NH ₂ .

  In this case, high cytotoxicity can be expected.

In indoline derivative or a pharmacologically acceptable salt thereof represented by the above general formula (I), X, -C (= O) NH ( CH 2) 2 Y or -NHC (= O) _CH 2 NH ₂ and Y is preferably a 4-aminophenyl group.

  In this case, high cytotoxicity can be expected.

In indoline derivative or a pharmacologically acceptable salt thereof represented by the above general formula (I), X, -C (= O) NH ( CH 2) 2 Y or -NHC (= O) _CH 2 NH ₂ , Y is a 4-aminophenyl group, and R ¹ is preferably a hydrogen atom or a methoxy group.

  In this case, high cytotoxicity can be expected.

In indoline derivative or a pharmacologically acceptable salt thereof represented by the above general formula (I), X, -C (= O) NH ( CH 2) 2 Y or -NHC (= O) _CH 2 NH ₂ , Y is a 4-aminophenyl group, and R ¹ is preferably a hydrogen atom.

  In this case, high cytotoxicity can be expected.

  The present invention also provides a cytotoxic agent containing the indoline derivative represented by the above general formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient.

  The indoline derivative of the present invention or a pharmacologically acceptable salt thereof has high cytotoxicity to cancer cells, and thus can be used as a cytotoxic agent.

［式中、Ｘは、−ＣＯ_２Ｍｅ、−ＣＯ_２Ｅｔ、−Ｃ（＝Ｏ）ＮＨ（ＣＨ_２）_２Ｙ又は−ＮＨＣ（＝Ｏ）Ｒ^２を表し、Ｙは、一個の水素原子がＮＨ_２で置換されたフェニル基を表し、Ｒ^１は、水素原子又はメトキシ基を表し、Ｒ^２は、メチル基、エチル基又は−ＣＨ_２ＮＨ_２を表す。］The indoline derivative of the present invention is characterized by being represented by the following general formula (I).

[In the _{formula, X, -CO 2 Me, -CO 2 Et} , represent _{-C (= O) NH (CH} 2) 2 Y or ^{-NHC (= O) R 2,} Y is the one hydrogen atom It represents a phenyl group substituted with NH ₂ , R ¹ represents a hydrogen atom or a methoxy group, and R ² represents a methyl group, an ethyl group or —CH ₂ NH ₂ . ]

  The following terms used in this specification have the following definitions unless otherwise specified.

  Me means a methyl group, and Et means an ethyl group.

The "phenyl group which one hydrogen atom is substituted with NH _2" means a 2-aminophenyl group, 3-aminophenyl group or a 4-aminophenyl group.

  Specific examples of preferable compounds of the indoline derivative represented by the above general formula (I) are shown in Table 1, but the present invention is not limited thereto.

  The compounds listed in Table 1 also include their pharmacologically acceptable salts.

  The indoline derivative represented by the above general formula (I) (hereinafter, indoline derivative (I)) contains a conformational isomer, a rotational isomer, a tautomer, an optical isomer, a diastereomer or an epimer. In some cases, either isomer or a mixture is included in the indoline derivative (I). Furthermore, when the indoline derivative (I) has an optical isomer, the optical isomer resolved from the racemate is also included in the indoline derivative (I).

  The indoline derivative (I) of the present invention includes a prodrug of the indoline derivative (I) or a pharmaceutically acceptable salt thereof. The prodrug of the indoline derivative (I) is a compound that is enzymatically or chemically converted into the indoline derivative (I) in vivo. The active substance of the prodrug of the indoline derivative (I) is the indoline derivative (I), but the prodrug itself of the indoline derivative (I) may have activity.

  Examples of the prodrug of the indoline derivative (I) include compounds in which the hydroxy group of the indoline derivative (I) is esterified, carbonated, carbamated, alkylated, phosphorylated or borated. These compounds can be synthesized from the indoline derivative (I) or a synthetic intermediate thereof according to a known method.

  Examples of the prodrug of the indoline derivative (I) include a compound in which the amino group of the indoline derivative (I) is carbamate or amidated. These compounds can be synthesized from the indoline derivative (I) or a synthetic intermediate thereof according to a known method.

  In addition, prodrugs of the indoline derivative (I) have been described in publicly known literature (“Development of Pharmaceuticals”, Hirokawa Shoten, 1990, Vol. 7, pp. 163-198, and Progress in Medicine, Vol. 5, 1985, p. 2157 to 2161), it may be changed to an indoline derivative (I) under physiological conditions.

The indoline derivative (I) may be labeled with an isotope, and examples of the labeled isotope include ² H, ³ H, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁵ O, ¹⁷ O, ¹⁸ O. And / or ¹²⁵ I.

  Examples of the "pharmacologically acceptable salt" of the indoline derivative (I) include inorganic acid salts such as hydrochloride, sulfate, nitrate, hydrobromide, hydroiodide or phosphate, and the like. Oxalate, malonate, citrate, fumarate, lactate, malate, succinate, tartrate, acetate, trifluoroacetate, maleate, gluconate, benzoate, Ascorbate, glutarate, mandelate, phthalate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, camphorsulfonate, aspartate, glutamate or Organic acid salts such as cinnamic acid salts, triethylamine salts, diisopropylethylamine salts, ethanolamine salts, morpholine salts, piperidine salts or dicyclohexylamites With salts with organic bases such as salts, salts with basic amino acids such as arginine or lysine, or with inorganic bases such as lithium salts, sodium salts, potassium salts, magnesium salts, calcium salts, ammonium salts, aluminum salts or zinc salts Salt can be mentioned.

  The indoline derivative (I) or a pharmaceutically acceptable salt thereof may be an anhydride or may form a solvate such as a hydrate. Here, the solvate is preferably a pharmaceutically acceptable solvate. The pharmaceutically acceptable solvate may be either a hydrate or a non-hydrate, but a hydrate is preferred. Examples of the solvent that constitutes the solvate include alcohol solvents such as methanol, ethanol, and n-propanol, N, N-dimethylformamide, dimethylsulfoxide, and water.

  The indoline derivative (I) can be produced by an appropriate method based on the characteristics derived from the basic skeleton and the kinds of substituents. The starting materials and reagents used for the production of these compounds can be generally purchased, or can be produced by a known method or a method analogous thereto.

  The indoline derivative (I) and the intermediates and starting materials used for the production thereof can be isolated and purified by the known means. Known means for isolation and purification include, for example, solvent extraction, recrystallization or chromatography.

  When the indoline derivative (I) contains an optical isomer or a stereoisomer, each isomer can be obtained as a single compound by a known method. Known methods include, for example, crystallization, enzyme resolution or chiral chromatography.

  In each reaction of the production methods described below, when the raw material compound has a hydroxy group, an amino group or a carboxyl group, a protecting group may be introduced into these groups, and the protecting group is optionally removed after the reaction. By protecting, the target compound can be obtained.

  Examples of the hydroxy-protecting group include a trityl group, a tetrahydropyranyl group, an aralkyl group having 7 to 10 carbon atoms (for example, a benzyl group) or a substituted silyl group (for example, a trimethylsilyl group, a triethylsilyl group or tert-butyldimethyl). Silyl group).

  Examples of the amino group-protecting group include an alkylcarbonyl group having 2 to 6 carbon atoms (eg, acetyl group), a trifluoroacetyl group, a benzoyl group, an alkyloxycarbonyl group having 2 to 8 carbon atoms (eg, tert-butoxy group). A carbonyl group or a benzyloxycarbonyl group), an aralkyl group having 7 to 10 carbon atoms (for example, a benzyl group), or a phthaloyl group.

  Examples of the protective group for the carboxyl group include an alkyl group having 1 to 6 carbon atoms (eg, methyl group, ethyl group or tert-butyl group) or an aralkyl group having 7 to 10 carbon atoms (eg, benzyl group).

  Deprotection of the protecting group depends on the type of the protecting group, but is performed according to a known method (for example, Greene, TW, "Greene's Protective Groups in Organic Synthesis", Wiley-Interscience) or a method analogous thereto. be able to.

［式中、Ａは、−ＣＯ_２Ｍｅ、−ＣＯ_２Ｅｔ、−Ｃ（＝Ｏ）ＮＨ（ＣＨ_２）_２Ｙ、−ＮＨＣ（＝Ｏ）Ｍｅ又は−ＮＨＣ（＝Ｏ）Ｅｔを表し、Ｍｅは、メチル基を表し、Ｅｔは、エチル基を表し、Ｂｎは、ベンジル基を表し、それ以外の各記号は、上記定義に同じである。］The indoline derivative (Ia) can be obtained, for example, by a debenzylation reaction of a benzyl-protected indoline derivative (II) as shown in Scheme 1.

[In the _{formula, A, -CO 2 Me, -CO 2 Et} , -C (= O) NH (CH 2) 2 Y, represents a -NHC (= O) Me, or -NHC (= O) Et, Me Represents a methyl group, Et represents an ethyl group, Bn represents a benzyl group, and other symbols are the same as defined above. ]

  The debenzylation reaction can be performed according to a known method (for example, Greene, TW, “Greene's Protective Groups in Organic Synthesis”, Wiley-Interscience) or a method analogous thereto.

［式中、Ｐｒｏは、ベンジル基以外の保護基を表し、それ以外の各記号は、上記定義に同じである。］The indoline derivative (Ib) can be obtained, for example, by a debenzylation reaction of a benzyl-protected indoline derivative (II-a) and a subsequent deprotection reaction of Pro, as shown in Scheme 2.

[In the formula, Pro represents a protective group other than a benzyl group, and other symbols are the same as defined above. ]

  The debenzylation reaction and the deprotection reaction of Pro can be performed according to a known method (for example, Greene, TW, "Greene's Protective Groups in Organic Synthesis", Wiley-Interscience) or a method analogous thereto. ..

［式中、各記号は、上記定義に同じである。］The benzyl-protected indoline derivative (II-a) can be obtained by, for example, an electrophilic reaction of an aniline derivative (III) as shown in Scheme 3.

[In the formula, each symbol is the same as defined above. ]

  Examples of the electrophile used in the electrophilic reaction include acid chlorides or mixed acid anhydrides of glycine in which the amino group is protected by Pro, and mixed acid anhydrides of N- (tert-butoxycarbonyl) glycine are preferable. ..

  The amount of the electrophile used in the electrophilic reaction is preferably 0.5 to 10 equivalents, and more preferably 1 to 3 equivalents with respect to the aniline derivative (III).

  The reaction solvent used in the electrophilic reaction is appropriately selected according to the type of reagent used, but is not particularly limited as long as it does not inhibit the reaction, and examples thereof include N, N-dimethylformamide and N, N-dimethyl. Acetamide, aprotic polar solvent such as N-methyl-2-pyrrolidone or dimethyl sulfoxide, ether solvent such as diethyl ether, tetrahydrofuran, dimethoxyethane or 1,4-dioxane, ester solvent such as ethyl acetate or propyl acetate, Examples of the solvent include chlorine-based solvents such as dichloromethane, chloroform or 1,2-dichloroethane, nitrile-based solvents such as acetonitrile or propionitrile, and mixed solvents thereof, but chlorine-based solvents such as dichloromethane, chloroform or 1,2-dichloroethane or the like. N, - dimethylformamide, N, N- dimethylacetamide, aprotic polar solvents such as N- methyl-2-pyrrolidone or dimethyl sulfoxide is preferable.

  The electrophilic reaction may optionally use a base. Examples of the base to be used include inorganic bases such as sodium hydride, sodium hydrogen carbonate or potassium carbonate, organic bases such as triethylamine, diisopropylethylamine, 4-dimethylaminopyridine or pyridine, or a mixture thereof, but triethylamine, diisopropyl. Organic bases such as ethylamine, 4-dimethylaminopyridine or pyridine or mixtures thereof are preferred.

  The amount of the base used for the electrophilic reaction is preferably 0.5 to 20 equivalents, more preferably 1 to 5 equivalents, based on the aniline derivative (III).

  The reaction temperature of the electrophilic reaction is preferably −40 ° C. to 200 ° C., more preferably −20 ° C. to 150 ° C.

  The reaction time of the electrophilic reaction is appropriately selected according to the conditions such as the reaction temperature, but is preferably 30 minutes to 30 hours.

  The concentration of the aniline derivative (III) used in the electrophilic reaction at the start of the reaction is preferably 1 mmol / L to 1 mol / L.

  The electrophile can be purchased or can be synthesized by a known method or a method analogous thereto.

［式中、各記号は、上記定義に同じである。］The benzyl-protected indoline derivative (II-b) can be obtained, for example, by a condensation reaction between a carboxylic acid derivative (IV) and an amine derivative (V) as shown in Scheme 4.

[In the formula, each symbol is the same as defined above. ]

  The amount of the amine derivative (V) used in the condensation reaction is preferably 0.5 to 10 equivalents, and more preferably 1 to 3 equivalents relative to the carboxylic acid derivative (IV).

  Examples of the condensing agent used in the condensation reaction include ethyl chloroformate, oxalyl chloride, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride, and hexafluorophosphoric acid 2- (7-aza-1H-benzotriazole- 1-yl) -1,1,3,3-tetramethyluronium, benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate, O- (benzotriazol-1-yl) -N, N , N ′, N′-tetramethyluronium hexafluorophosphate, 1H-benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate or 2-methyl-6-nitrobenzoic anhydride. 1-Ethyl-3- (3-dimethylaminopropyl) carbodihydrochloride Mido, 2- (7-aza-1H-benzotriazol-1-yl) -1,1,3,3-tetramethyluronium hexafluorophosphate or benzotriazol-1-yloxytris (dimethyl hexafluorophosphate) Amino) phosphonium is preferred.

  The amount of the condensing agent used in the condensation reaction is preferably 0.1 to 100 equivalents, more preferably 0.3 to 30 equivalents, based on the carboxylic acid derivative (IV).

  The reaction solvent used in the condensation reaction is appropriately selected depending on the type of reagent used, but is not particularly limited as long as it does not inhibit the reaction, and examples thereof include N, N-dimethylformamide and N, N-dimethylacetamide. Aprotic polar solvent such as N-methyl-2-pyrrolidone or dimethyl sulfoxide, ether solvent such as diethyl ether, tetrahydrofuran, dimethoxyethane or 1,4-dioxane, ester solvent such as ethyl acetate or propyl acetate, dichloromethane , A chlorine-based solvent such as chloroform or 1,2-dichloroethane, a nitrile-based solvent such as acetonitrile or propionitrile or a mixed solvent thereof, but a chlorine-based solvent such as dichloromethane, chloroform or 1,2-dichloroethane or N. , N Dimethylformamide, N, N- dimethylacetamide, aprotic polar solvents such as N- methyl-2-pyrrolidone or dimethyl sulfoxide is preferable.

  A base may be used in the condensation reaction, if desired. Examples of the base to be used include inorganic bases such as sodium hydride, sodium hydrogen carbonate or potassium carbonate, organic bases such as triethylamine, diisopropylethylamine, 4-dimethylaminopyridine or pyridine, or a mixture thereof, but triethylamine, diisopropyl. Organic bases such as ethylamine, 4-dimethylaminopyridine or pyridine or mixtures thereof are preferred.

  The amount of the base used in the condensation reaction is preferably 0.5 to 20 equivalents, more preferably 1 to 5 equivalents, based on the carboxylic acid derivative (IV).

  The reaction temperature of the condensation reaction is preferably −40 ° C. to 200 ° C., more preferably −20 ° C. to 150 ° C.

  The reaction time of the condensation reaction is appropriately selected according to the conditions such as the reaction temperature, but is preferably 30 minutes to 30 hours.

  The concentration of the carboxylic acid derivative (IV) used in the condensation reaction at the start of the reaction is preferably 1 mmol / L to 1 mol / L.

  The amine derivative (V) can be purchased or can be synthesized by a known method or a method analogous thereto.

［式中、各記号は、上記定義に同じである。］The aniline derivative (III) can be obtained by, for example, a hydrolysis reaction of a benzyl-protected indoline derivative (VI) as shown in Scheme 5.

[In the formula, each symbol is the same as defined above. ]

  The hydrolysis reaction can be performed according to a known method (for example, Greene, TW, “Greene's Protective Groups in Organic Synthesis”, Wiley-Interscience) or a method analogous thereto.

［式中、Ｂは、−ＣＯ_２Ｍｅ又は−ＣＯ_２Ｅｔを表し、それ以外の各記号は、上記定義に同じである。］The carboxylic acid derivative (IV) can be obtained, for example, by a hydrolysis reaction of a benzyl-protected indoline derivative (II-c) as shown in Scheme 6, respectively.

Wherein, B represents a _-CO 2 Me or _-CO 2 Et, other each symbol is defined the same as above. ]

  The hydrolysis reaction can be performed according to a known method (for example, Greene, TW, “Greene's Protective Groups in Organic Synthesis”, Wiley-Interscience) or a method analogous thereto.

［式中、Ｒ^３は、メチル基、エチル基又はトリフルオロメチル基を表し、それ以外の各記号は、上記定義に同じである。］The benzyl-protected indoline derivative (II-d / VI) can be prepared, for example, by the deprotection reaction of Pro of the protected indoline derivative (VII) and the subsequent condensation reaction with the indole derivative (VIII) as shown in Scheme 7. Obtainable.

[In the formula, R ³ represents a methyl group, an ethyl group or a trifluoromethyl group, and other symbols are the same as defined above. ]

  The deprotection reaction of Pro can be performed according to a known method (for example, Greene, TW, “Greene's Protective Groups in Organic Synthesis”, Wiley-Interscience) or a method analogous thereto.

  The amount of the indole derivative (VIII) used in the condensation reaction is preferably 0.5 to 10 equivalents, more preferably 1 to 3 equivalents, relative to the protected indoline derivative (VII).

  Examples of the condensing agent used in the condensation reaction include ethyl chloroformate, oxalyl chloride, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride, and hexafluorophosphoric acid 2- (7-aza-1H-benzotriazole- 1-yl) -1,1,3,3-tetramethyluronium, benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate, O- (benzotriazol-1-yl) -N, N , N ′, N′-tetramethyluronium hexafluorophosphate, 1H-benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate or 2-methyl-6-nitrobenzoic anhydride. Hydrochloric acid 1-ethyl-3- (3-dimethylaminopropyl) carbodii 2- (7-aza-1H-benzotriazol-1-yl) -1,1,3,3-tetramethyluronium hexafluorophosphate or benzotriazol-1-yloxytris (dimethyl hexafluorophosphate) Amino) phosphonium is preferred.

  The amount of the condensing agent used in the condensation reaction is preferably 0.1 to 100 equivalents, more preferably 0.3 to 30 equivalents, relative to the indole derivative (VIII).

  The reaction solvent used in the condensation reaction is appropriately selected depending on the type of reagent used, but is not particularly limited as long as it does not inhibit the reaction, and examples thereof include N, N-dimethylformamide and N, N-dimethylacetamide. Aprotic polar solvent such as N-methyl-2-pyrrolidone or dimethyl sulfoxide, ether solvent such as diethyl ether, tetrahydrofuran, dimethoxyethane or 1,4-dioxane, ester solvent such as ethyl acetate or propyl acetate, dichloromethane Examples include chlorine-based solvents such as chloroform and 1,2-dichloroethane, nitrile-based solvents such as acetonitrile and propionitrile, and mixed solvents thereof. Chlorine-based solvents such as dichloromethane, chloroform or 1,2-dichloroethane, or N. , N Dimethylformamide, N, N- dimethylacetamide, aprotic polar solvents such as N- methyl-2-pyrrolidone or dimethyl sulfoxide is preferable.

  A base may be used in the condensation reaction, if desired. Examples of the base to be used include inorganic bases such as sodium hydride, sodium hydrogen carbonate or potassium carbonate, organic bases such as triethylamine, diisopropylethylamine, 4-dimethylaminopyridine or pyridine, or a mixture thereof, but triethylamine, diisopropyl. Organic bases such as ethylamine, 4-dimethylaminopyridine or pyridine or mixtures thereof are preferred.

  The amount of the base used in the condensation reaction is preferably 0.5 to 20 equivalents, more preferably 1 to 5 equivalents, relative to the protected indoline derivative (VII).

  The reaction temperature of the condensation reaction is preferably −40 ° C. to 200 ° C., more preferably −20 ° C. to 150 ° C.

  The reaction time of the condensation reaction is appropriately selected according to the conditions such as the reaction temperature, but is preferably 30 minutes to 30 hours.

  The concentration of the protected indoline derivative (VII) used in the condensation reaction at the start of the reaction is preferably 1 mmol / L to 1 mol / L.

［式中、各記号は、上記定義に同じである。］The benzyl-protected indoline derivative (II-c) can be obtained, for example, by the deprotection reaction of Pro of the protected indoline derivative (IX) and the subsequent condensation reaction with the indole derivative (VIII) as shown in Scheme 8. You can

[In the formula, each symbol is the same as defined above. ]

  The deprotection reaction of Pro can be performed according to a known method (for example, Greene, TW, “Greene's Protective Groups in Organic Synthesis”, Wiley-Interscience) or a method analogous thereto.

  The amount of the indole derivative (VIII) used in the condensation reaction is preferably 0.5 to 10 equivalents, more preferably 1 to 3 equivalents relative to the protected indoline derivative (IX).

  Examples of the condensing agent used in the condensation reaction include ethyl chloroformate, oxalyl chloride, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride, and hexafluorophosphoric acid 2- (7-aza-1H-benzotriazole- 1-yl) -1,1,3,3-tetramethyluronium, benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate, O- (benzotriazol-1-yl) -N, N , N ′, N′-tetramethyluronium hexafluorophosphate, 1H-benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate or 2-methyl-6-nitrobenzoic anhydride. Hydrochloric acid 1-ethyl-3- (3-dimethylaminopropyl) carbodii 2- (7-aza-1H-benzotriazol-1-yl) -1,1,3,3-tetramethyluronium hexafluorophosphate or benzotriazol-1-yloxytris (dimethyl hexafluorophosphate) Amino) phosphonium is preferred.

  The amount of the condensing agent used in the condensation reaction is preferably 0.1 to 100 equivalents, more preferably 0.3 to 30 equivalents, relative to the indole derivative (VIII).

  The reaction solvent used in the condensation reaction is appropriately selected depending on the type of reagent used, but is not particularly limited as long as it does not inhibit the reaction, and examples thereof include N, N-dimethylformamide and N, N-dimethylacetamide. Aprotic polar solvent such as N-methyl-2-pyrrolidone or dimethyl sulfoxide, ether solvent such as diethyl ether, tetrahydrofuran, dimethoxyethane or 1,4-dioxane, ester solvent such as ethyl acetate or propyl acetate, dichloromethane , A chlorine-based solvent such as chloroform or 1,2-dichloroethane, a nitrile-based solvent such as acetonitrile or propionitrile or a mixed solvent thereof, but a chlorine-based solvent such as dichloromethane, chloroform or 1,2-dichloroethane or N. , N Dimethylformamide, N, N- dimethylacetamide, aprotic polar solvents such as N- methyl-2-pyrrolidone or dimethyl sulfoxide is preferable.

  A base may be used in the condensation reaction, if desired. Examples of the base to be used include inorganic bases such as sodium hydride, sodium hydrogen carbonate or potassium carbonate, organic bases such as triethylamine, diisopropylethylamine, 4-dimethylaminopyridine or pyridine, or a mixture thereof, but triethylamine, diisopropyl. Organic bases such as ethylamine, 4-dimethylaminopyridine or pyridine or mixtures thereof are preferred.

  The amount of the base used in the condensation reaction is preferably 0.5 to 20 equivalents, more preferably 1 to 5 equivalents, relative to the protected indoline derivative (IX).

  The reaction temperature of the condensation reaction is preferably −40 ° C. to 200 ° C., more preferably −20 ° C. to 150 ° C.

  The reaction time of the condensation reaction is appropriately selected according to the conditions such as the reaction temperature, but is preferably 30 minutes to 30 hours.

  The concentration of the protected indoline derivative (IX) used in the condensation reaction at the start of the reaction is preferably 1 mmol / L to 1 mol / L.

  The protected indoline derivative (IX) can be synthesized by the method described in Examples below, a known method, or a method analogous thereto.

  The indole derivative (VIII) can be purchased or can be synthesized by a known method or a method analogous thereto.

［式中、各記号は、上記定義に同じである。］The protected indoline derivative (VII) can be obtained, for example, by the electrophilic reaction of the aniline derivative (X) as shown in Scheme 9.

[In the formula, each symbol is the same as defined above. ]

  Examples of the electrophile used in the electrophilic reaction include acetic anhydride, acetyl chloride, propionic anhydride, propionyl chloride, trifluoroacetic anhydride, or trifluoroacetyl chloride.

  The amount of the electrophile used in the electrophilic reaction is preferably 0.5 to 10 equivalents, and more preferably 1 to 3 equivalents with respect to the aniline derivative (X).

  The reaction solvent used in the electrophilic reaction is appropriately selected according to the type of reagent used, but is not particularly limited as long as it does not inhibit the reaction, and examples thereof include N, N-dimethylformamide and N, N-dimethyl. Acetamide, aprotic polar solvent such as N-methyl-2-pyrrolidone or dimethyl sulfoxide, ether solvent such as diethyl ether, tetrahydrofuran, dimethoxyethane or 1,4-dioxane, ester solvent such as ethyl acetate or propyl acetate, Examples of the solvent include chlorine-based solvents such as dichloromethane, chloroform or 1,2-dichloroethane, nitrile-based solvents such as acetonitrile or propionitrile, and mixed solvents thereof, but chlorine-based solvents such as dichloromethane, chloroform or 1,2-dichloroethane or the like. N, - dimethylformamide, N, N- dimethylacetamide, aprotic polar solvents such as N- methyl-2-pyrrolidone or dimethyl sulfoxide is preferable.

  The electrophilic reaction may optionally use a base. Examples of the base to be used include inorganic bases such as sodium hydride, sodium hydrogen carbonate or potassium carbonate, organic bases such as triethylamine, diisopropylethylamine, 4-dimethylaminopyridine or pyridine, or a mixture thereof, but triethylamine, diisopropyl. Organic bases such as ethylamine, 4-dimethylaminopyridine or pyridine or mixtures thereof are preferred.

  The amount of the base used for the electrophilic reaction is preferably 0.5 to 20 equivalents, more preferably 1 to 5 equivalents, based on the aniline derivative (X).

  The reaction temperature of the electrophilic reaction is preferably −40 ° C. to 200 ° C., more preferably −20 ° C. to 150 ° C.

  The reaction time of the electrophilic reaction is appropriately selected according to the conditions such as the reaction temperature, but is preferably 30 minutes to 30 hours.

  The concentration of the aniline derivative (X) used in the electrophilic reaction at the start of the reaction is preferably 1 mmol / L to 1 mol / L.

  The electrophile used in the electrophilic reaction can be purchased or can be synthesized by a known method or a method analogous thereto.

  The aniline derivative (X) can be purchased or can be synthesized by a known method or a method analogous thereto.

  Further, the cytotoxic agent of the present invention is characterized by containing an indoline derivative (I) or a pharmacologically acceptable salt thereof as an active ingredient.

  The “cytotoxic agent” means a composition containing a compound that causes cell death by impairing cell function or inhibiting cell growth or a pharmacologically acceptable salt thereof as an active ingredient. The indoline derivative (I) or a pharmacologically acceptable salt thereof is considered to act on the cell division mechanism by the replication of DNA, and is not particularly limited as long as it has the mechanism. For example, cancer cells It is preferably used as a cytotoxic agent against immune cells, bacteria, fungi, pathogenic protozoa, yeasts or viruses.

  The indoline derivative (I) or a pharmaceutically acceptable salt thereof is administered to a mammal (eg, mouse, rat, hamster, rabbit, cat, dog, monkey, cow, sheep or human), particularly human. In some cases, it can be used as a useful cytotoxic agent.

  The indoline derivative (I) or a pharmaceutically acceptable salt thereof may be orally or parenterally administered as a cytotoxic agent to the above mammals as it is or in combination with a pharmaceutically acceptable carrier. You can

  Examples of “parenteral administration” include injection administration, nasal administration, pulmonary administration, transdermal administration, sublingual administration and rectal administration, and injection administration is preferred.

  "Injection administration" means systemically or locally administering a cytotoxic agent by injection or drip, and the administration site includes, for example, intravenous, intramuscular, intraperitoneal, and subcutaneous.

  The cytotoxic agent containing the indoline derivative (I) or a pharmacologically acceptable salt thereof as an active ingredient is a known method generally used in the technical field of formulation (for example, Remington's Pharmaceutical Science, latest edition). , Mark Publishing Company, Easton, USA). The above-mentioned cytotoxic agents are, if necessary, excipients, binders, disintegrating agents, lubricants, sweeteners, surfactants, suspending agents, emulsifiers, which are usually used in the field of formulation. Additives such as colorants, preservatives, fragrances, corrigents, stabilizers, thickeners, buffers, isotonicity agents and fluidity promoters can be added in appropriate amounts. Examples of the pharmacologically acceptable carrier include these additives.

  The cytotoxic agent containing the indoline derivative (I) or a pharmacologically acceptable salt thereof as an active ingredient is a disease in which improvement of pathological condition or amelioration of symptoms can be expected based on the mechanism of action, for example, cancer, autoimmune disease. Alternatively, it can be used as a therapeutic drug for infectious diseases.

  "Cancer" is a disease caused by a cell population that undergoes uncontrolled abnormal growth, and includes, for example, pharyngeal cancer, laryngeal cancer, tongue cancer, non-small cell lung cancer, breast cancer, esophageal cancer, gastric cancer, colon cancer, uterus. Cancer, endometrial cancer, ovarian cancer, liver cancer, pancreatic cancer, gallbladder cancer, cholangiocarcinoma, renal cancer, renal pelvic and ureteral cancer, bladder cancer, prostate cancer, malignant melanoma, thyroid cancer, neuroosteosarcoma, chondrosarcoma, Rhabdomyosarcoma, angiosarcoma, fibrosarcoma, glioma, leukemia, malignant lymphoma, neuroblastoma, myeloma or brain tumor.

  The "cancer cell" is a cell that causes cancer, and includes, for example, cells of pharyngeal cancer, cells of laryngeal cancer, cells of tongue cancer, cells of non-small cell lung cancer, cells of breast cancer, cells of esophageal cancer. , Gastric cancer cells, colon cancer cells, uterine cancer cells, endometrial cancer cells, ovarian cancer cells, liver cancer cells, pancreatic cancer cells, gallbladder cancer cells, cholangiocarcinoma cells, kidney cancer cells Cells, renal pelvic and ureteral cancer cells, bladder cancer cells, prostate cancer cells, malignant melanoma cells, thyroid cancer cells, neuroosteosarcoma cells, chondrosarcoma cells, rhabdomyosarcoma cells, hemangiosarcoma Cells, fibrosarcoma cells, glioma cells, leukemia cells, malignant lymphoma cells, neuroblastoma cells, myeloma cells or brain tumor cells.

  The “immune cell” is a cell that causes an autoimmune disease, and includes, for example, B cells, T cells, dendritic cells, macrophages, NK cells, neutrophils, eosinophils or basophils.

  “Autoimmune disease” is a disease caused by an abnormal immune response to substances or cells in the living body, and examples thereof include active chronic hepatitis, Addison's disease, ankylosing spondylitis, antiphospholipid syndrome, arthritis, atopic disease. Allergy, Behcet's disease, Cardiomyopathy, Celiac disease, Corgan's syndrome, Cold coagulosis, Crohn's disease, Cushing's syndrome, Dermatomyositis, Lupus erythematosus, Erythema, Fibromyalgia, Glomerulonephritis, Goodpasture's syndrome, Graft pair Host disease, Graves' disease, Guillain-Barre syndrome, Hashimoto's disease, idiopathic adrenal atrophy, idiopathic pulmonary fibrosis, IgA nephropathy, inflammatory bowel disease, insulin-dependent diabetes mellitus, juvenile arthritis, Lambert-Eaton myasthenic syndrome, Lichen planus, lupus hepatitis, lupus, lymphopenia, Meniere's disease, mixed connective tissue disease, multiple sclerosis Myasthenia gravis, pernicious anemia, polyendocrine hypofunction syndrome, primary biliary cirrhosis, psoriasis, psoriatic arthritis, Raynaud's phenomenon, Reiter's syndrome, rheumatoid arthritis, Schmidt syndrome, scleroderma, Sjogren's syndrome, Stiffman's syndrome, mating Sensitive ophthalmitis, systemic lupus erythematosus, Takayasu arteritis, temporal arteritis, hyperthyroidism, insulin resistance type B, type I diabetes, ulcerative colitis, uveitis, vitiligo or Wegener's granulomatosis.

  "Infectious disease" is a disease caused by infection with bacteria, fungi, pathogenic protozoa, yeasts or viruses, etc., and as the bacteria, fungi, pathogenic protozoa, yeasts or viruses that cause the above-mentioned infections, For example, Staphylococcus, Streptococcus, Enterococcus, Corynebacterium, Bacillus, Listeria, Peptococcus, Peptostreptococcus, Clostridium, Eubacterium, Propionibacterium, Lactobacillus, Neisseria, Blanchamella, Haemophilus, Bordetella, Escherichia coli, Citrotro Bacter, Salmonella, Shigella, Klebsiella, Enterobacter, Serratia, Hafnia, Proteus, Morganella, Providencia, Yersinia, Campylobacter, Vibrio, Aeromonas, Pseudomonas, Xanthomonas, Acinetobacter, Fu Bobacterium, brucella, legionella, bayronella, bacteroides, fusobacterium, mycoplasma, rickettsia, chlamydia, aspergillus, waliptococcus, candida, mucor, sporotrichum, dermatophyte, malaria parasite, dysentery amoeba, vaginal trichomonas, pneumochicus carini, echinococcus Streptococcus pneumoniae, Haemophilus influenzae, Mycobacterium tuberculosis, Tetanus, Adenovirus, Herpesvirus, Papillomavirus, AIDS virus, Filovirus, Japanese encephalitis virus, Rabies virus, Poliovirus, Rhinovirus, Influenza virus, Hepatitis B virus or C type Hepatitis virus is mentioned.

The cytotoxicity of the indoline derivative (I) or a pharmaceutically acceptable salt thereof can be evaluated using an in vitro test. Examples of in vitro tests include trypan blue dye exclusion method, lactate dehydrogenase (LDH) activity method, [ ³ H] thymidine incorporation method, propidium iodide, which measures the number of dead cells or viable cells after compound treatment. ATP using a nuclear staining method, a method using a dye such as MTT, MTS and WST (Current Protocol in Toxicology, 2000, 2.6.1-2.6.27) or CellTiter-Glow (product of Promega) The amount of viable cells after the compound treatment is measured by the MTS method, and the cytotoxicity value at that time is determined by the 50% action concentration (EC ₅₀ ) is preferable. For the in vitro test, mouse lymphocytic leukemia-derived L1210 cells and the like are used.

  Hereinafter, the present invention will be described in detail with reference to Examples and Reference Examples, but the present invention is not limited thereto.

；
８−（ベンジルオキシ）−２−ナフトエ酸メチル誘導体：

In addition, about the compound used for the synthesis | combination of an Example compound and a synthetic method is not described, the compound synthesize | combined with the commercially available or well-known method or the method according to it was used. The solvent name shown in the NMR data indicates the solvent used for the measurement. The 400 MHz NMR spectrum was measured using a JNM-AL400 type nuclear magnetic resonance apparatus (manufactured by JEOL Ltd.) or a JNM-ECS400 type nuclear magnetic resonance apparatus (manufactured by JEOL Ltd.). The chemical shift is represented by δ (unit: ppm) with respect to tetramethylsilane, and signals are s (single line), d (double line), t (triplet line), q (quad line), and m, respectively. (Multiple line), br (wide), dd (double double line), dt (double triple line), ddd (double double double line), dq (double quadruple line) or tt (triple triple line). Triple line). In the proton NMR spectrum, those that cannot be confirmed broadly such as OH and NH protons are not described in the data. The molecular weight is measured by Agilent Technologies 1200 Series, 6130A (manufactured by Agilent Technologies) (hereinafter, LC / MS-1) or Agilent Technologies 1260 Infinity II Series, 6130B (manufactured by Agilent Technologies) (hereinafter, LC / MS-2). Was measured by an electrospray ionization (hereinafter, ESI) method. The analysis conditions for LC / MS-1 and LC / MS-2 will be described later. The following abbreviations are used in the description of the following examples and reference examples.
Ac: acetyl group; Boc: tert-butoxycarbonyl group; Bn: benzyl group; Me: methyl group;
4- (benzyloxy) -6-bromonaphthalene derivative:

;
Methyl 8- (benzyloxy) -2-naphthoate derivative:

アルゴン雰囲気下、（４−（ベンジルオキシ）−６−ブロモナフタレン−２−イル）カルバミン酸 ｔｅｒｔ−ブチル（１００ｍｇ，０．２３ｍｍｏｌ）のＮ，Ｎ−ジメチルホルムアミド（以下、ＤＭＦ）溶液（１．０ｍＬ）に４−ジメチルアミノピリジン（以下、ＤＭＡＰ）（２８ｍｇ，０．２３ｍｍｏｌ）、二炭酸ジ−ｔｅｒｔ−ブチル（以下、Ｂｏｃ_２Ｏ）（１２５ｍｇ，０．５８ｍｍｏｌ）を加え、６０℃で３時間撹拌した。反応液を濃縮することにより得られた残渣をシリカゲルカラムクロマトグラフィー（酢酸エチル／ｎ−ヘキサン＝１／６）により精製し、表題化合物（以下、参考例１の化合物）を無色固体として得た（１０５ｍｇ，収率８６％）。
^１Ｈ−ＮＭＲ（４００ＭＨｚ，ＣＤＣｌ_３）δ：１．３９（１８Ｈ，ｓ），５．２３（２Ｈ，ｓ），６．６８（１Ｈ，ｄ，Ｊ＝１．６Ｈｚ），７．１９（１Ｈ，ｄ，Ｊ＝０．８Ｈｚ），７．３８−７．５１（５Ｈ，ｍ），７．５８（１Ｈ，ｄｄ，Ｊ＝８．８，１．６Ｈｚ），７．６４（１Ｈ，ｄ，Ｊ＝８．８Ｈｚ），８．４５（１Ｈ，ｄ，Ｊ＝１．６Ｈｚ）．
ＥＳＩ−ＭＳ（ｍ／ｚ）：５２８［Ｍ＋Ｈ］^＋．（ＬＣ／ＭＳ−１）Reference Example 1 Synthesis of 4- (benzyloxy) -6-bromonaphthalene derivative:

Under argon atmosphere, tert-butyl (4- (benzyloxy) -6-bromonaphthalen-2-yl) carbamate (100 mg, 0.23 mmol) in N, N-dimethylformamide (hereinafter, DMF) solution (1.0 mL ) To 4-dimethylaminopyridine (hereinafter, DMAP) (28 mg, 0.23 mmol) and di-tert-butyl dicarbonate (hereinafter, Boc ₂ O) (125 mg, 0.58 mmol), and stirred at 60 ° C. for 3 hours. did. The residue obtained by concentrating the reaction solution was purified by silica gel column chromatography (ethyl acetate / n-hexane = 1/6) to obtain the title compound (hereinafter, compound of Reference Example 1) as a colorless solid ( 105 mg, yield 86%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 1.39 (18 H, s), 5.23 (2 H, s), 6.68 (1 H, d, J = 1.6 Hz), 7.19 (1 H). , D, J = 0.8 Hz), 7.38-7.51 (5H, m), 7.58 (1H, dd, J = 8.8, 1.6 Hz), 7.64 (1H, d, J = 8.8 Hz), 8.45 (1H, d, J = 1.6 Hz).
ESI-MS (m / z): 528 [M + H] ⁺ . (LC / MS-1)

アルゴン雰囲気下、参考例１の化合物（１．１９ｇ，２．２５ｍｍｏｌ）の脱水ＤＭＦ溶液（２０ｍＬ）にＮ−ホルミルサッカリン（１．４３ｇ，６．７５ｍｍｏｌ）、酢酸パラジウム（１５ｍｇ，０．０７ｍｍｏｌ）、Ｘａｎｔｐｈｏｓ（５８ｍｇ，０．１０ｍｍｏｌ）、フッ化カリウム（６５４ｍｇ，１１．３ｍｍｏｌ）を加え、簡易脱気（反応系を真空ポンプで吸引し、泡出たせ、アルゴンガスを吹き込む操作を３回繰り返す操作）し、８０℃で終夜撹拌した。さらにトリエチルアミン（０．４７ｍＬ，３．３８ｍｍｏｌ）、脱水メタノール（１．０ｍＬ）を加え、室温で３時間撹拌した。反応液をセライトろ過し、続いて濃縮することにより得られた残渣をシリカゲルカラムクロマトグラフィー（酢酸エチル／ｎ−ヘキサン＝１／１０）により精製し、表題化合物（以下、参考例２の化合物）を無色アモルファスとして得た（４１８ｍｇ，収率３７％）。
^１Ｈ−ＮＭＲ（４００ＭＨｚ，ＣＤＣｌ_３）δ：１．３８（１８Ｈ，ｓ），３．９８（３Ｈ，ｓ），５．２９（２Ｈ，ｓ），６．６９（１Ｈ，ｄ，Ｊ＝２．０Ｈｚ），７．１９−７．２５（１Ｈ，ｍ），７．３１−７．３７（１Ｈ，ｍ），７．４２（２Ｈ，ｔ，Ｊ＝７．２Ｈｚ），７．５１（２Ｈ，ｄ，Ｊ＝７．２Ｈｚ），７．８１（１Ｈ，ｄ，Ｊ＝８．８Ｈｚ），８．０９（１Ｈ，ｄｄ，Ｊ＝８．８，２．０Ｈｚ），９．０５（１Ｈ，ｄ，Ｊ＝１．２Ｈｚ）．
ＥＳＩ−ＭＳ（ｍ／ｚ）：５０８［Ｍ＋Ｈ］^＋．（ＬＣ／ＭＳ−１）Reference Example 2 Synthesis of methyl 8- (benzyloxy) -2-naphthoate derivative:

N-formylsaccharin (1.43 g, 6.75 mmol), palladium acetate (15 mg, 0.07 mmol) in a dehydrated DMF solution (20 mL) of the compound of Reference Example 1 (1.19 g, 2.25 mmol) under an argon atmosphere. Xantphos (58 mg, 0.10 mmol) and potassium fluoride (654 mg, 11.3 mmol) were added, and simple degassing (operation of sucking the reaction system with a vacuum pump, causing bubbles, and blowing argon gas three times) And stirred at 80 ° C. overnight. Furthermore, triethylamine (0.47 mL, 3.38 mmol) and dehydrated methanol (1.0 mL) were added, and the mixture was stirred at room temperature for 3 hours. The reaction solution is filtered through Celite, and the residue obtained by subsequent concentration is purified by silica gel column chromatography (ethyl acetate / n-hexane = 1/10) to give the title compound (hereinafter, referred to as the compound of Reference Example 2). Obtained as a colorless amorphous substance (418 mg, yield 37%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 1.38 (18 H, s), 3.98 (3 H, s), 5.29 (2 H, s), 6.69 (1 H, d, J = 2). 0.0 Hz), 7.19-7.25 (1H, m), 7.31-7.37 (1H, m), 7.42 (2H, t, J = 7.2Hz), 7.51 (2H). , D, J = 7.2 Hz), 7.81 (1 H, d, J = 8.8 Hz), 8.09 (1 H, dd, J = 8.8, 2.0 Hz), 9.05 (1 H, d, J = 1.2 Hz).
ESI-MS (m / z): 508 [M + H] ⁺ . (LC / MS-1)

アルゴン雰囲気下、参考例２の化合物（７２２ｍｇ，１．４２ｍｍｏｌ）のジクロロメタン溶液（１０ｍＬ）にトリフルオロ酢酸（以下、ＴＦＡ）（０．２１８ｍＬ，２．８４ｍｍｏｌ）を加え、室温で２時間撹拌した。反応液に飽和炭酸水素ナトリウム水溶液を加え、クロロホルムで３回抽出し、無水硫酸ナトリウムで乾燥した。ろ過、続いて濃縮することにより得られた残渣を再沈殿法（クロロホルム／ｎ−ヘキサン）により精製し、表題化合物（以下、参考例３の化合物）を無色固体として得た（４８０ｍｇ，収率８３％）。
^１Ｈ−ＮＭＲ（４００ＭＨｚ，ＣＤＣｌ_３）δ：１．５５（９Ｈ，ｓ），３．９５（３Ｈ，ｓ），５．２８（２Ｈ，ｓ），６．６７（１Ｈ，ｓ），７．０９（１Ｈ，ｄ，Ｊ＝１．６Ｈｚ），７．３６−７．４６（４Ｈ，ｍ），７．５３（２Ｈ，ｄ，Ｊ＝７．２Ｈｚ），７．７０（１Ｈ，ｄ，Ｊ＝８．４Ｈｚ），８．０２（１Ｈ，ｄ，Ｊ＝８．４，１．６Ｈｚ），８．９５（１Ｈ，ｄ，Ｊ＝１．２Ｈｚ）．
ＥＳＩ−ＭＳ（ｍ／ｚ）：４０８［Ｍ＋Ｈ］^＋．（ＬＣ／ＭＳ−１）Reference Example 3 Synthesis of methyl 8- (benzyloxy) -6-((tert-butoxycarbonyl) amino) -2-naphthoate:

Under an argon atmosphere, trifluoroacetic acid (hereinafter, TFA) (0.218 mL, 2.84 mmol) was added to a dichloromethane solution (10 mL) of the compound of Reference Example 2 (722 mg, 1.42 mmol), and the mixture was stirred at room temperature for 2 hours. A saturated aqueous solution of sodium hydrogen carbonate was added to the reaction solution, extracted three times with chloroform, and dried over anhydrous sodium sulfate. The residue obtained by filtration and subsequent concentration was purified by the reprecipitation method (chloroform / n-hexane) to obtain the title compound (hereinafter, compound of Reference Example 3) as a colorless solid (480 mg, yield 83). %).
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 1.55 (9 H, s), 3.95 (3 H, s), 5.28 (2 H, s), 6.67 (1 H, s), 7. 09 (1H, d, J = 1.6Hz), 7.36-7.46 (4H, m), 7.53 (2H, d, J = 7.2Hz), 7.70 (1H, d, J) = 8.4 Hz), 8.02 (1H, d, J = 8.4, 1.6 Hz), 8.95 (1H, d, J = 1.2 Hz).
ESI-MS (m / z): 408 [M + H] ⁺ . (LC / MS-1)

アルゴン雰囲気下、参考例３の化合物（４８０ｍｇ，１．１８ｍｍｏｌ）のテトラヒドロフラン（以下、ＴＨＦ）−メタノール混合溶液（５．０ｍＬ／５．０ｍＬ）にＮ−ヨードスクシンイミド（３９８ｍｇ，１．７７ｍｍｏｌ）、ｐ−トルエンスルホン酸一水和物（１１２ｍｇ，０．５９ｍｍｏｌ）を−７８℃で加え、１時間撹拌し、さらに０℃で３時間撹拌した。反応液に飽和チオ硫酸ナトリウム水溶液を加え、クロロホルムで３回抽出し、無水硫酸ナトリウムで乾燥した。ろ過、続いて濃縮することにより得られた残渣をシリカゲルカラムクロマトグラフィー（クロロホルム／ｎ−ヘキサン＝１／２）により精製し、表題化合物（以下、参考例４の化合物）を無色固体として得た（５７６ｍｇ，収率９２％）。
^１Ｈ−ＮＭＲ（４００ＭＨｚ，ＣＤＣｌ_３）δ：１．５８（９Ｈ，ｓ），３．９６（３Ｈ，ｓ），５．３２（２Ｈ，ｓ），７．３７−７．５８（４Ｈ，ｍ），７．５７（２Ｈ，ｄ，Ｊ＝６．８Ｈｚ），８．０５（１Ｈ，ｄ，Ｊ＝８．８Ｈｚ），８．１０（１Ｈ，ｄｄ，Ｊ＝９．２，２．０Ｈｚ），８．１５（１Ｈ，ｂｒｓ），８．９４（１Ｈ，ｄ，Ｊ＝２．０Ｈｚ）．
ＥＳＩ−ＭＳ（ｍ／ｚ）：５３４［Ｍ＋Ｈ］^＋．（ＬＣ／ＭＳ−１）Reference Example 4 Synthesis of methyl 8- (benzyloxy) -6-((tert-butoxycarbonyl) amino) -5-iodo-2-naphthoate:

Under an argon atmosphere, N-iodosuccinimide (398 mg, 1.77 mmol) was added to a tetrahydrofuran (hereinafter, THF) -methanol mixed solution (5.0 mL / 5.0 mL) of the compound of Reference Example 3 (480 mg, 1.18 mmol), p. -Toluenesulfonic acid monohydrate (112 mg, 0.59 mmol) was added at -78 ° C, stirred for 1 hour, and further stirred at 0 ° C for 3 hours. A saturated aqueous sodium thiosulfate solution was added to the reaction solution, extracted three times with chloroform, and dried over anhydrous sodium sulfate. The residue obtained by filtration and subsequent concentration was purified by silica gel column chromatography (chloroform / n-hexane = 1/2) to obtain the title compound (hereinafter, compound of Reference Example 4) as a colorless solid ( 576 mg, yield 92%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 1.58 (9H, s), 3.96 (3H, s), 5.32 (2H, s), 7.37-7.58 (4H, m). ), 7.57 (2H, d, J = 6.8Hz), 8.05 (1H, d, J = 8.8Hz), 8.10 (1H, dd, J = 9.2, 2.0Hz) , 8.15 (1H, brs), 8.94 (1H, d, J = 2.0 Hz).
ESI-MS (m / z): 534 [M + H] ⁺ . (LC / MS-1)

アルゴン雰囲気下、参考例４の化合物（５７６ｍｇ，１．０８ｍｍｏｌ）の脱水ＤＭＦ溶液（３．０ｍＬ）にヨウ化テトラブチルアンモニウム（２０ｍｇ，０．０５４ｍｍｏｌ）、水素化ナトリウム（５５重量％鉱油分散物）（１１８ｍｇ，２．７０ｍｍｏｌ）を加え、０℃で１時間撹拌した。さらに１，３−ジクロロプロペン（０．２６６ｍＬ，３．２４ｍｍｏｌ）を加え、０℃で３時間撹拌した。さらに室温で終夜撹拌した。反応液に水を加え、クロロホルムで３回抽出し、無水硫酸ナトリウムで乾燥した。ろ過、続いて濃縮することにより得られた残渣をシリカゲルカラムクロマトグラフィー（クロロホルム／ｎ−ヘキサン＝１／１）により精製し、表題化合物（以下、参考例５の化合物）を黄色アモルファスとして得た（５８９ｍｇ，収率９０％）。
^１Ｈ−ＮＭＲ（４００ＭＨｚ，ＣＤＣｌ_３）δ：１．３０（９Ｈ，ｓ），３．７５−３．８１（１Ｈ，ｍ），４．００（３Ｈ，ｓ），４．４５−４．６２（１Ｈ，ｍ），５．２３−５．３４（２Ｈ，ｍ），５．９３−６．１０（２Ｈ，ｍ），７．３５−７．５２（６Ｈ，ｍ），８．１５（１Ｈ，ｄ，Ｊ＝８．８Ｈｚ），８．２５（１Ｈ，ｄｄ，Ｊ＝８．８，２．８Ｈｚ），９．０２−９．０８（１Ｈ，ｍ）．
ＥＳＩ−ＭＳ（ｍ／ｚ）：６０８［Ｍ＋Ｈ］^＋．（ＬＣ／ＭＳ−１）Reference Example 5 Synthesis of methyl 8- (benzyloxy) -6-((tert-butoxycarbonyl) (3-chloroallyl) amino) -5-iodo-2-naphthoate:

Under an argon atmosphere, tetrabutylammonium iodide (20 mg, 0.054 mmol) and sodium hydride (55 wt% mineral oil dispersion) were added to a dehydrated DMF solution (3.0 mL) of the compound of Reference Example 4 (576 mg, 1.08 mmol). (118 mg, 2.70 mmol) was added, and the mixture was stirred at 0 ° C for 1 hr. Further, 1,3-dichloropropene (0.266 mL, 3.24 mmol) was added, and the mixture was stirred at 0 ° C. for 3 hours. Further, the mixture was stirred at room temperature overnight. Water was added to the reaction solution, extracted three times with chloroform, and dried over anhydrous sodium sulfate. The residue obtained by filtration and subsequent concentration was purified by silica gel column chromatography (chloroform / n-hexane = 1/1) to obtain the title compound (hereinafter, compound of Reference Example 5) as a yellow amorphous ( 589 mg, yield 90%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 1.30 (9H, s), 3.75-3.81 (1H, m), 4.00 (3H, s), 4.45-4.62. (1H, m), 5.23-5.34 (2H, m), 5.93-6.10 (2H, m), 7.35-7.52 (6H, m), 8.15 (1H). , D, J = 8.8 Hz), 8.25 (1H, dd, J = 8.8, 2.8 Hz), 9.02-9.08 (1H, m).
ESI-MS (m / z): 608 [M + H] ⁺ . (LC / MS-1)

参考例５の化合物（１４０ｍｇ，０．２３ｍｍｏｌ）の脱水ベンゼン溶液（５．０ｍＬ）をアルゴンガスで３０分間バブリングした。この溶液にトリストリメチルシラン（０．０７２ｍＬ，０．２１ｍｍｏｌ）、アゾビスイソブチロニトリル（以下、ＡＩＢＮ）（９ｍｇ，０．０５２ｍｍｏｌ）を加え、３時間加熱還流した。反応液を濃縮し、得られた残渣をシリカゲルカラムクロマトグラフィー（クロロホルム／ｎ−ヘキサン＝１／１）により精製し、表題化合物（以下、参考例６の化合物）を黄色アモルファスとして得た（９６ｍｇ，収率８３％）。
^１Ｈ−ＮＭＲ（４００ＭＨｚ，ＣＤＣｌ_３）δ：１．５８（９Ｈ，ｓ），３．４５（１Ｈ，ｔ，Ｊ＝１０．８Ｈｚ），３．９０（１Ｈ，ｄｄ，Ｊ＝１０．８，２．４Ｈｚ），３．９５（３Ｈ，ｓ），３．９２−３．９９（１Ｈ，ｍ），４．１１−４．１８（１Ｈ，ｍ），４．２４−４．３０（１Ｈ，ｍ），５．２９（２Ｈ，ｓ），７．３５−７．５０（５Ｈ，ｍ），７．５２−７．５８（１Ｈ，ｍ），７．６４（１Ｈ，ｄ，Ｊ＝９．６Ｈｚ），８．０６（１Ｈ，ｄｄ，Ｊ＝９．６，１．６Ｈｚ），９．００−９．０５（１Ｈ，ｍ）．
ＥＳＩ−ＭＳ（ｍ／ｚ）：４８２［Ｍ＋Ｈ］^＋．（ＬＣ／ＭＳ−１）Reference Example 6 3- (tert-Butyl) 7-methyl-5- (benzyloxy) -1- (chloromethyl) -1,2-dihydro-3H-benzo [e] indole-3,7-dicarboxy Rat synthesis:

A dehydrated benzene solution (5.0 mL) of the compound of Reference Example 5 (140 mg, 0.23 mmol) was bubbled with argon gas for 30 minutes. Tristrimethylsilane (0.072 mL, 0.21 mmol) and azobisisobutyronitrile (hereinafter, AIBN) (9 mg, 0.052 mmol) were added to this solution, and the mixture was heated under reflux for 3 hours. The reaction mixture was concentrated, and the obtained residue was purified by silica gel column chromatography (chloroform / n-hexane = 1/1) to give the title compound (hereinafter, compound of Reference Example 6) as a yellow amorphous substance (96 mg, Yield 83%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 1.58 (9 H, s), 3.45 (1 H, t, J = 10.8 Hz), 3.90 (1 H, dd, J = 10.8, 2.4 Hz), 3.95 (3H, s), 3.92-3.99 (1H, m), 4.11-4.18 (1H, m), 4.24-4.30 (1H, m), 5.29 (2H, s), 7.35-7.50 (5H, m), 7.52-7.58 (1H, m), 7.64 (1H, d, J = 9.m). 6 Hz), 8.06 (1H, dd, J = 9.6, 1.6 Hz), 9.00-9.05 (1H, m).
ESI-MS (m / z): 482 [M + H] ⁺ . (LC / MS-1)

アルゴン雰囲気下、参考例６の化合物（９６ｍｇ，０．２０ｍｍｏｌ）に４Ｎ塩化水素−ジオキサン溶液（３．０ｍＬ）を加え、室温で３時間撹拌した。反応液を濃縮し、得られた残渣にＤＭＦ（３．０ｍＬ）、５，６−ジメトキシ−１Ｈ−インドール−２−カルボン酸（６６ｍｇ，０．３０ｍｍｏｌ）、塩酸１−エチル−３−（３−ジメチルアミノプロピル）カルボジイミド（以下、ＥＤＣＩ）（１１５ｍｇ，０．６０ｍｍｏｌ）を加え、室温で終夜撹拌した。反応液に水を加え、クロロホルムで３回抽出し、無水硫酸ナトリウムで乾燥した。ろ過、続いて濃縮することにより得られた残渣をシリカゲルカラムクロマトグラフィー（クロロホルム／ｎ−ヘキサン＝１／１）により精製し、表題化合物（以下、参考例７の化合物）を黄色アモルファスとして得た（８０ｍｇ，収率６８％）。
^１Ｈ−ＮＭＲ（４００ＭＨｚ，ＣＤＣｌ_３）δ：３．４８（１Ｈ，ｔ，Ｊ＝１０．８Ｈｚ），３．６３−３．６８（１Ｈ，ｍ），３．７８（３Ｈ，ｓ），３．９４（３Ｈ，ｓ），３．９７（３Ｈ，ｓ），４．０８−４．１６（１Ｈ，ｍ），４．６２−４．７０（１Ｈ，ｍ），４．７６−４．８１（１Ｈ，ｍ），５．９２（２Ｈ，ｓ），６．８５（１Ｈ，ｓ），７．０３（１Ｈ，ｄ，Ｊ＝１．２Ｈｚ），７．０９（１Ｈ，ｓ），７．３２−７．４５（３Ｈ，ｍ），７．５５（２Ｈ，ｄ，Ｊ＝７．６Ｈｚ），７．７０（１Ｈ，ｄ，Ｊ＝８．８Ｈｚ），８．１１（１Ｈ，ｄｄ，Ｊ＝８．８，２．０Ｈｚ），８．２５（１Ｈ，ｓ），９．０５（１Ｈ，ｄ，Ｊ＝１．２Ｈｚ），９．５２（１Ｈ，ｓ）．
ＥＳＩ−ＭＳ（ｍ／ｚ）：５８５［Ｍ＋Ｈ］^＋．（ＬＣ／ＭＳ−１）Reference Example 7 5- (Benzyloxy) -1- (chloromethyl) -3- (5,6-dimethoxy-1H-indole-2-carbonyl) -2,3-dihydro-1H-benzo [e] indole Synthesis of methyl-7-carboxylate:

Under an argon atmosphere, a 4N hydrogen chloride-dioxane solution (3.0 mL) was added to the compound of Reference Example 6 (96 mg, 0.20 mmol), and the mixture was stirred at room temperature for 3 hours. The reaction solution was concentrated, and DMF (3.0 mL), 5,6-dimethoxy-1H-indole-2-carboxylic acid (66 mg, 0.30 mmol), and 1-ethyl-3- (3-hydrochloride) were added to the obtained residue. Dimethylaminopropyl) carbodiimide (hereinafter, EDCI) (115 mg, 0.60 mmol) was added, and the mixture was stirred at room temperature overnight. Water was added to the reaction solution, extracted three times with chloroform, and dried over anhydrous sodium sulfate. The residue obtained by filtration and subsequent concentration was purified by silica gel column chromatography (chloroform / n-hexane = 1/1) to obtain the title compound (hereinafter, compound of Reference Example 7) as a yellow amorphous ( 80 mg, yield 68%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 3.48 (1 H, t, J = 10.8 Hz), 3.63-3.68 (1 H, m), 3.78 (3 H, s), 3 .94 (3H, s), 3.97 (3H, s), 4.08-4.16 (1H, m), 4.62-4.70 (1H, m), 4.76-4.81. (1H, m), 5.92 (2H, s), 6.85 (1H, s), 7.03 (1H, d, J = 1.2Hz), 7.09 (1H, s), 7. 32-7.45 (3H, m), 7.55 (2H, d, J = 7.6Hz), 7.70 (1H, d, J = 8.8Hz), 8.11 (1H, dd, J) = 8.8, 2.0 Hz), 8.25 (1H, s), 9.05 (1H, d, J = 1.2 Hz), 9.52 (1H, s).
ESI-MS (m / z): 585 [M + H] ⁺ . (LC / MS-1)

アルゴン雰囲気下、参考例７の化合物（３１ｍｇ，０．０５３ｍｍｏｌ）のＴＨＦ−メタノール混合溶液（６．０ｍＬ／２．０ｍＬ）に１０重量％パラジウム／カーボン（３１ｍｇ、５０重量％含水）、２５重量％ギ酸アンモニウム水溶液（０．４００ｍＬ）を加え、室温で７０分間撹拌した。反応液をセライトろ過し、水を加え、クロロホルムで３回抽出し、無水硫酸ナトリウムで乾燥した。ろ過、続いて濃縮することにより得られた残渣をシリカゲルカラムクロマトグラフィー（メタノール／クロロホルム＝１／１００→１／５０）により精製し、表題化合物（以下、実施例１の化合物）を黄色固体として得た（８ｍｇ，収率３０％）。
^１Ｈ−ＮＭＲ（４００ＭＨｚ，ＤＭＳＯ−ｄ_６）δ：３．７９（３Ｈ，ｓ），３．８１（３Ｈ，ｓ），３．８４−３．９０（１Ｈ，ｍ），３．９１（３Ｈ，ｓ），４．０３（１Ｈ，ｄｄ，Ｊ＝１１．２，２．８Ｈｚ），４．２２−４．２８（１Ｈ，ｍ），４．５６（１Ｈ，ｄｄ，Ｊ＝１１．２，２．０Ｈｚ），４．８０（１Ｈ，ｔ，Ｊ＝１１．２Ｈｚ），６．９６（１Ｈ，ｓ），７．１０（１Ｈ，ｄ，Ｊ＝２．４Ｈｚ），７．１５（１Ｈ，ｓ），７．９３−７．９７（２Ｈ，ｍ），８．０７（１Ｈ，ｓ），８．８１（１Ｈ，ｓ），１０．９０（１Ｈ，ｂｒｓ），１１．５２（１Ｈ，ｂｒｓ）．
ＥＳＩ−ＭＳ（ｍ／ｚ）：４９５［Ｍ＋Ｈ］^＋．（ＬＣ／ＭＳ−１）(Example 1) 1- (chloromethyl) -3- (5,6-dimethoxy-1H-indole-2-carbonyl) -5-hydroxy-2,3-dihydro-1H-benzo [e] indole-7- Synthesis of methyl carboxylate:

Under an argon atmosphere, a THF-methanol mixed solution (6.0 mL / 2.0 mL) of the compound of Reference Example 7 (31 mg, 0.053 mmol) contained 10 wt% palladium / carbon (31 mg, 50 wt% water content), 25 wt%. Aqueous ammonium formate solution (0.400 mL) was added, and the mixture was stirred at room temperature for 70 minutes. The reaction mixture was filtered through Celite, water was added, and the mixture was extracted 3 times with chloroform and dried over anhydrous sodium sulfate. The residue obtained by filtration and subsequent concentration was purified by silica gel column chromatography (methanol / chloroform = 1/100 → 1/50) to obtain the title compound (hereinafter, compound of Example 1) as a yellow solid. (8 mg, yield 30%).
¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 3.79 (3H, s), 3.81 (3H, s), 3.84-3.90 (1H, m), 3.91 (3H). , S), 4.03 (1H, dd, J = 11.2, 2.8 Hz), 4.22-4.28 (1H, m), 4.56 (1H, dd, J = 11.2, 2.0 Hz), 4.80 (1 H, t, J = 11.2 Hz), 6.96 (1 H, s), 7.10 (1 H, d, J = 2.4 Hz), 7.15 (1 H, s), 7.93-7.97 (2H, m), 8.07 (1H, s), 8.81 (1H, s), 10.90 (1H, brs), 11.52 (1H, brs). ).
ESI-MS (m / z): 495 [M + H] ⁺ . (LC / MS-1)

アルゴン雰囲気下、参考例７の化合物（３６ｍｇ，０．０６２ｍｍｏｌ）のメタノール懸濁液（４．０ｍＬ）に、１Ｎ水酸化ナトリウム水溶液（１．０ｍＬ）を加え、６０℃で２日間撹拌した。反応液に１Ｎ塩酸を加え、クロロホルムで３回抽出し、無水硫酸ナトリウムで乾燥した。ろ過、続いて濃縮することにより得られた残渣をシリカゲルカラムクロマトグラフィー（メタノール／クロロホルム＝１／５０）により精製し、表題化合物（以下、参考例８の化合物）を黄色アモルファスとして得た（１４ｍｇ，収率３７％）。
^１Ｈ−ＮＭＲ（４００ＭＨｚ，ＣＤＣｌ_３）δ：３．４９−３．５８（２Ｈ，ｍ），３．９４（３Ｈ，ｓ），３．９５（３Ｈ，ｓ），４．１１−４．１８（２Ｈ，ｍ），５．０２−５．０６（２Ｈ，ｍ），５．３０−５．３９（２Ｈ，ｍ），６．８７（１Ｈ，ｓ），７．００−７．１４（２Ｈ，ｍ），７．３４−７．６０（５Ｈ，ｍ），８．０９−８．２３（２Ｈ，ｍ），８．９４（１Ｈ，ｓ），９．１０（１Ｈ，ｓ），９．５９（１Ｈ，ｂｒｓ）．
ＥＳＩ−ＭＳ（ｍ／ｚ）：５６９［Ｍ−Ｈ］⁻．（ＬＣ／ＭＳ−１）Reference Example 8 5- (Benzyloxy) -1- (chloromethyl) -3- (5,6-dimethoxy-1H-indole-2-carbonyl) -2,3-dihydro-1H-benzo [e] indole Synthesis of -7-carboxylic acid:

Under an argon atmosphere, a 1N aqueous sodium hydroxide solution (1.0 mL) was added to a methanol suspension (4.0 mL) of the compound of Reference Example 7 (36 mg, 0.062 mmol), and the mixture was stirred at 60 ° C for 2 days. 1N Hydrochloric acid was added to the reaction solution, extracted three times with chloroform, and dried over anhydrous sodium sulfate. The residue obtained by filtration and subsequent concentration was purified by silica gel column chromatography (methanol / chloroform = 1/50) to obtain the title compound (hereinafter, compound of Reference Example 8) as a yellow amorphous substance (14 mg, Yield 37%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 3.49-3.58 (2H, m), 3.94 (3H, s), 3.95 (3H, s), 4.11-4.18. (2H, m), 5.02-5.06 (2H, m), 5.30-5.39 (2H, m), 6.87 (1H, s), 7.00-7.14 (2H , M), 7.34-7.60 (5H, m), 8.09-8.23 (2H, m), 8.94 (1H, s), 9.10 (1H, s), 9. 59 (1H, brs).
ESI-MS (m / z): 569 [MH] ⁻ . (LC / MS-1)

アルゴン雰囲気下、参考例８の化合物（１２ｍｇ，０．０２１ｍｍｏｌ）のジクロロメタン溶液（１．０ｍＬ）に、４−（２−アミノエチル）アニリン（４ｍｇ，０．０３２ｍｍｏｌ）、トリエチルアミン（０．００５ｍＬ，０．０３２ｍｍｏｌ）、ヘキサフルオロリン酸ベンゾトリアゾール−１−イルオキシトリス(ジメチルアミノ)ホスホニウム（１４ｍｇ，０．０３２ｍｍｏｌ）を加え、室温で終夜撹拌した。反応液に飽和炭酸水素ナトリウム水溶液を加え、クロロホルムで３回抽出し、無水硫酸ナトリウムで乾燥した。ろ過、続いて濃縮することにより得られた残渣をシリカゲルカラムクロマトグラフィー（メタノール／クロロホルム＝１／５０）により精製し、表題化合物（以下、参考例９の化合物）を茶色アモルファスとして得た（１１ｍｇ，収率４８％）。
^１Ｈ−ＮＭＲ（４００ＭＨｚ，ＣＤＣｌ_３）δ：２．８４（２Ｈ，ｔ，Ｊ＝６．８Ｈｚ），３．４７（２Ｈ，ｔ，Ｊ＝６．８Ｈｚ），３．５８（２Ｈ，ｂｒｓ），３．６５−３．７０（２Ｈ，ｍ），３．９２（３Ｈ，ｓ），３．９５（３Ｈ，ｓ），４．０８−４．１５（１Ｈ，ｍ），４．６５−４．７１（１Ｈ，ｍ），４．７７−４．８２（１Ｈ，ｍ），５．２７−５．３６（２Ｈ，ｍ），６．２９−６．３４（１Ｈ，ｍ），６．５７−６．６１（２Ｈ，ｍ），６．８７（１Ｈ，ｓ），６．９２−７．０５（４Ｈ，ｍ），７．１０（１Ｈ，ｓ），７．３６−７．４７（３Ｈ，ｍ），７．５４（２Ｈ，ｄ，Ｊ＝７．６Ｈｚ），７．７１（１Ｈ，ｄ，Ｊ＝８．４Ｈｚ），７．９０（１Ｈ，ｄｄ，Ｊ＝８．８，１．６Ｈｚ），８．６０（１Ｈ，ｓ），９．４０（１Ｈ，ｂｒｓ）．
ＥＳＩ−ＭＳ（ｍ／ｚ）：６８９［Ｍ＋Ｈ］^＋．（ＬＣ／ＭＳ−１）Reference Example 9 N- (4-aminophenethyl) -5- (benzyloxy) -1- (chloromethyl) -3- (5,6-dimethoxy-1H-indole-2-carbonyl) -2,3- Synthesis of dihydro-1H-benzo [e] indole-7-carboxamide:

Under an argon atmosphere, 4- (2-aminoethyl) aniline (4 mg, 0.032 mmol) and triethylamine (0.005 mL, 0) were added to a dichloromethane solution (1.0 mL) of the compound of Reference Example 8 (12 mg, 0.021 mmol). 0.032 mmol) and benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate (14 mg, 0.032 mmol) were added, and the mixture was stirred at room temperature overnight. A saturated aqueous solution of sodium hydrogen carbonate was added to the reaction solution, extracted three times with chloroform, and dried over anhydrous sodium sulfate. The residue obtained by filtration and subsequent concentration was purified by silica gel column chromatography (methanol / chloroform = 1/50) to obtain the title compound (hereinafter, compound of Reference Example 9) as a brown amorphous substance (11 mg, Yield 48%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 2.84 (2H, t, J = 6.8 Hz), 3.47 (2H, t, J = 6.8 Hz), 3.58 (2H, brs). , 3.65-3.70 (2H, m), 3.92 (3H, s), 3.95 (3H, s), 4.08-4.15 (1H, m), 4.65-4. .71 (1H, m), 4.77-4.82 (1H, m), 5.27-5.36 (2H, m), 6.29-6.34 (1H, m), 6.57. -6.61 (2H, m), 6.87 (1H, s), 6.92-7.05 (4H, m), 7.10 (1H, s), 7.36-7.47 (3H) , M), 7.54 (2H, d, J = 7.6 Hz), 7.71 (1H, d, J = 8.4 Hz), 7.90 (1H, dd, J = 8.8, 1. 6Hz), 8.60 (1H, s , 9.40 (1H, brs).
ESI-MS (m / z): 689 [M + H] ⁺ . (LC / MS-1)

参考例７の化合物の代わりに、参考例９の化合物（７ｍｇ，０．０１０ｍｍｏｌ）を用いて、それ以外は実施例１と同様の方法にて合成を行い、表題化合物（以下、実施例２の化合物）を白色固体（５ｍｇ，収率７３％）として得た。
^１Ｈ−ＮＭＲ（４００ＭＨｚ，ＤＭＳＯ−ｄ_６）δ：２．６３−２．６９（２Ｈ，ｍ），３．３８−３．４８（２Ｈ，ｍ），３．６４−３．６９（１Ｈ，ｍ），３．７８（３Ｈ，ｓ），３．８４（３Ｈ，ｓ），３．８０−３．８６（１Ｈ，ｍ），３．９８−４．０３（１Ｈ，ｍ），４．２２（１Ｈ，ｂｒｓ），４．５１（１Ｈ，ｄ，Ｊ＝１．６Ｈｚ），４．７５−４．８５（２Ｈ，ｍ），６．４５−６．４９（２Ｈ，ｍ），６．８６−７．１１（５Ｈ，ｍ），７．８４−７．９１（２Ｈ，ｍ），８．００（１Ｈ，ｓ），８．６３−８．６８（２Ｈ，ｍ），１０．６９（１Ｈ，ｓ），１１．４７（１Ｈ，ｄ，Ｊ＝１．６Ｈｚ）．
ＥＳＩ−ＭＳ（ｍ／ｚ）：５９９［Ｍ＋Ｈ］^＋．（ＬＣ／ＭＳ−１）(Example 2) N- (4-aminophenethyl) -1- (chloromethyl) -3- (5,6-dimethoxy-1H-indole-2-carbonyl) -5-hydroxy-2,3-dihydro-1H Synthesis of Benzo [e] indole-7-carboxamide:

The compound of Reference Example 9 (7 mg, 0.010 mmol) was used instead of the compound of Reference Example 7, and the synthesis was performed in the same manner as in Example 1 except for that, and the title compound (hereinafter, referred to as Example 2 Compound) was obtained as a white solid (5 mg, yield 73%).
¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 2.63-2.69 (2H, m), 3.38-3.48 (2H, m), 3.64-3.69 (1H, m), 3.78 (3H, s), 3.84 (3H, s), 3.80-3.86 (1H, m), 3.98-4.03 (1H, m), 4.22. (1H, brs), 4.51 (1H, d, J = 1.6Hz), 4.75-4.85 (2H, m), 6.45-6.49 (2H, m), 6.86. -7.11 (5H, m), 7.84-7.91 (2H, m), 8.00 (1H, s), 8.63-8.68 (2H, m), 10.69 (1H) , S), 11.47 (1H, d, J = 1.6 Hz).
ESI-MS (m / z): 599 [M + H] ⁺ . (LC / MS-1)

アルゴン雰囲気下、７−（（（（９Ｈ−フルオレン−９−イル）メトキシ）カルボニル）アミノ）−５−（ベンジルオキシ）−１−（クロロメチル）−１，２−ジヒドロ−３Ｈ−ベンゾ［ｅ］インドール−３−カルボン酸 ｔｅｒｔ−ブチル（１０６ｍｇ，０．１６ｍｍｏｌ）のＴＨＦ溶液（３．０ｍＬ）に、１Ｍのフッ化テトラ−ｎ−ブチルアンモニウム（以下、ＴＢＡＦ）−ＴＨＦ溶液（０．２４７ｍＬ，０．２５ｍｍｏｌ）を加え、室温で３時間撹拌した。反応液に水を加え、クロロホルムで３回抽出し、無水硫酸ナトリウムで乾燥した。ろ過、続いて濃縮することにより得られた残渣をシリカゲルカラムクロマトグラフィー（クロロホルム）により粗精製し、得られた茶色アモルファスにジクロロメタン（２．０ｍＬ）、トリエチルアミン（０．０２７ｍＬ，０．１９ｍｍｏｌ）、無水酢酸（０．０１８ｍＬ，０．１９ｍｍｏｌ）を加え、室温で終夜撹拌した。反応液に飽和炭酸水素ナトリウム水溶液を加え、クロロホルムで３回抽出し、無水硫酸ナトリウムで乾燥した。ろ過、続いて濃縮することにより得られた残渣をシリカゲルカラムクロマトグラフィー（酢酸エチル／ｎ−ヘキサン＝１／４→１／２）により精製し、表題化合物（以下、参考例１０の化合物）を茶色アモルファスとして得た（４２ｍｇ，収率８５％）。
^１Ｈ−ＮＭＲ（４００ＭＨｚ，ＣＤＣｌ_３）δ：１．６０（９Ｈ，ｓ），２，２１（３Ｈ，ｓ），３．４３（１Ｈ，ｔ，Ｊ＝６．８Ｈｚ），３．８８−３．９８（２Ｈ，ｍ），４．０９−４．１５（１Ｈ，ｍ），４．２０−４．２７（１Ｈ，ｍ），５．２３（２Ｈ，ｓ），７．３５−７．６１（７Ｈ，ｍ），７．８６−８．０４（３Ｈ，ｍ）．
ＥＳＩ−ＭＳ（ｍ／ｚ）：４８１［Ｍ＋Ｈ］^＋．（ＬＣ／ＭＳ−１）Reference Example 10 Synthesis of tert-butyl 7-acetamido-5- (benzyloxy) -1- (chloromethyl) -1,2-dihydro-3H-benzo [e] indole-3-carboxylate:

7-((((9H-fluoren-9-yl) methoxy) carbonyl) amino) -5- (benzyloxy) -1- (chloromethyl) -1,2-dihydro-3H-benzo [e under an argon atmosphere. ] To a THF solution (3.0 mL) of tert-butyl indole-3-carboxylate (106 mg, 0.16 mmol), 1M tetra-n-butylammonium fluoride (hereinafter, TBAF) -THF solution (0.247 mL, 0.25 mmol) was added and the mixture was stirred at room temperature for 3 hours. Water was added to the reaction solution, extracted three times with chloroform, and dried over anhydrous sodium sulfate. The residue obtained by filtration and subsequent concentration was roughly purified by silica gel column chromatography (chloroform), and the brown amorphous obtained was dichloromethane (2.0 mL), triethylamine (0.027 mL, 0.19 mmol), anhydrous. Acetic acid (0.018 mL, 0.19 mmol) was added, and the mixture was stirred at room temperature overnight. A saturated aqueous solution of sodium hydrogen carbonate was added to the reaction solution, extracted three times with chloroform, and dried over anhydrous sodium sulfate. The residue obtained by filtration and subsequent concentration was purified by silica gel column chromatography (ethyl acetate / n-hexane = 1/4 → 1/2), and the title compound (compound of Reference Example 10 below) was brown. Obtained as an amorphous substance (42 mg, yield 85%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 1.60 (9 H, s), 2, 21 (3 H, s), 3.43 (1 H, t, J = 6.8 Hz), 3.88-3. .98 (2H, m), 4.09-4.15 (1H, m), 4.20-4.27 (1H, m), 5.23 (2H, s), 7.35-7.61. (7H, m), 7.86-8.04 (3H, m).
ESI-MS (m / z): 481 [M + H] ⁺ . (LC / MS-1)

参考例６の化合物の代わりに、参考例１０の化合物（４２ｍｇ，０．０８６ｍｍｏｌ）を用いて、それ以外は参考例７と同様の方法にて合成を行い、表題化合物（以下、参考例１１の化合物）を白色固体（９ｍｇ，収率１９％）として得た。
^１Ｈ−ＮＭＲ（４００ＭＨｚ，ＣＤＣｌ_３）δ：２．２４（３Ｈ，ｓ），３．４８（１Ｈ，ｔ，Ｊ＝６．８Ｈｚ），３．９８（６Ｈ，ｓ），３．９８−４．０２（１Ｈ，ｍ），４．１０−４．１７（１Ｈ，ｍ），４．６６−４．７４（１Ｈ，ｍ），４．７８−４．８２（１Ｈ，ｍ），５．２６−５．３５（２Ｈ，ｍ），６．９１（１Ｈ，ｓ），７．０５（１Ｈ，ｄ，Ｊ＝２．０Ｈｚ），７．１３（１Ｈ，ｓ），７．３７−７．４８（４Ｈ，ｍ），７．５６（２Ｈ，ｄ，Ｊ＝８．８Ｈｚ），７．７０（１Ｈ，ｄ，Ｊ＝８．８Ｈｚ），８．０３（１Ｈ，ｄｄ，Ｊ＝８．８，１．６Ｈｚ），８．１１（１Ｈ，ｄ，Ｊ＝１．６Ｈｚ），８．１９（１Ｈ，ｓ），９．３１−９．３５（１Ｈ，ｍ）．
ＥＳＩ−ＭＳ（ｍ／ｚ）：５８４［Ｍ＋Ｈ］^＋．（ＬＣ／ＭＳ−１）Reference Example 11 N- (5- (benzyloxy) -1- (chloromethyl) -3- (5,6-dimethoxy-1H-indole-2-carbonyl) -2,3-dihydro-1H-benzo [ e] Synthesis of indol-7-yl) acetamide:

Instead of the compound of Reference Example 6, the compound of Reference Example 10 (42 mg, 0.086 mmol) was used, and synthesis was carried out in the same manner as in Reference Example 7 except for that, to give the title compound (hereinafter referred to as Reference Example 11 Compound) was obtained as a white solid (9 mg, yield 19%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 2.24 (3H, s), 3.48 (1H, t, J = 6.8 Hz), 3.98 (6H, s), 3.98-4. .02 (1H, m), 4.10-4.17 (1H, m), 4.66-4.74 (1H, m), 4.78-4.82 (1H, m), 5.26. -5.35 (2H, m), 6.91 (1H, s), 7.05 (1H, d, J = 2.0Hz), 7.13 (1H, s), 7.37-7.48 (4H, m), 7.56 (2H, d, J = 8.8Hz), 7.70 (1H, d, J = 8.8Hz), 8.03 (1H, dd, J = 8.8,) 1.6 Hz), 8.11 (1H, d, J = 1.6 Hz), 8.19 (1H, s), 9.31-9.35 (1H, m).
ESI-MS (m / z): 584 [M + H] ⁺ . (LC / MS-1)

参考例７の化合物の代わりに、参考例１１の化合物（９ｍｇ，０．０１６ｍｍｏｌ）を用いて、反応時間を３０分間として、それ以外は実施例１と同様の方法にて合成を行い、表題化合物（以下、実施例３の化合物）を白色固体（６ｍｇ，収率７１％）として得た。
^１Ｈ−ＮＭＲ（４００ＭＨｚ，ＤＭＳＯ−ｄ_６）δ：２．１３（３Ｈ，ｓ），３．８２（３Ｈ，ｓ），３．８４（３Ｈ，ｓ），３．８４−３．９０（１Ｈ，ｍ），４．０６（１Ｈ，ｄｄ，Ｊ＝１０．８，２．８Ｈｚ），４．２２（１Ｈ，ｍ），４．５６（１Ｈ，ｄ，Ｊ＝１０．８Ｈｚ），４．７８（１Ｈ，ｔ，Ｊ＝１０．０Ｈｚ），７．００（１Ｈ，ｓ），７．０９（１Ｈ，ｓ），７．１８（１Ｈ，ｓ），７．７２（１Ｈ，ｄｄ，Ｊ＝９．２，２．０Ｈｚ），７．８３（１Ｈ，ｄ，Ｊ＝８．８Ｈｚ），７．９９（１Ｈ，ｓ），８．４６（１Ｈ，ｄ，Ｊ＝２．０Ｈｚ），１０．１２（１Ｈ，ｂｒｓ），１０．３８（１Ｈ，ｂｒｓ），１１．４９（１Ｈ，ｂｒｓ）．
ＥＳＩ−ＭＳ（ｍ／ｚ）：４９４［Ｍ＋Ｈ］^＋．（ＬＣ／ＭＳ−１）Example 3 (N- (1- (chloromethyl) -3- (5,6-dimethoxy-1H-indole-2-carbonyl) -5-hydroxy-2,3-dihydro-1H-benzo [e] Synthesis of indole-7-yl) acetamide:

The compound of Reference Example 11 (9 mg, 0.016 mmol) was used instead of the compound of Reference Example 7, the reaction time was 30 minutes, and synthesis was carried out in the same manner as in Example 1 except for that, and the title compound (Hereinafter, the compound of Example 3) was obtained as a white solid (6 mg, yield 71%).
¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 2.13 (3H, s), 3.82 (3H, s), 3.84 (3H, s), 3.84-3.90 (1H , M), 4.06 (1H, dd, J = 10.8, 2.8Hz), 4.22 (1H, m), 4.56 (1H, d, J = 10.8Hz), 4.78. (1H, t, J = 10.0 Hz), 7.00 (1H, s), 7.09 (1H, s), 7.18 (1H, s), 7.72 (1H, dd, J = 9) .2, 2.0 Hz), 7.83 (1 H, d, J = 8.8 Hz), 7.99 (1 H, s), 8.46 (1 H, d, J = 2.0 Hz), 10.12 (1H, brs), 10.38 (1H, brs), 11.49 (1H, brs).
ESI-MS (m / z): 494 [M + H] ⁺ . (LC / MS-1)

アルゴン雰囲気下、７−（（（（９Ｈ−フルオレン−９−イル）メトキシ）カルボニル）アミノ）−５−（ベンジルオキシ）−１−（クロロメチル）−１，２−ジヒドロ−３Ｈ−ベンゾ［ｅ］インドール−３−カルボン酸 ｔｅｒｔ−ブチル（７０２ｍｇ，１．０６ｍｍｏｌ）のＴＨＦ溶液（３．０ｍＬ）に、１ＭのＴＢＡＦ−ＴＨＦ溶液（１．５９ｍＬ，１．５９ｍｍｏｌ）を加え、室温で３時間撹拌した。反応液に水を加え、クロロホルムで３回抽出し、無水硫酸ナトリウムで乾燥した。ろ過、続いて濃縮することにより得られた残渣にジクロロメタン（３．０ｍＬ）、ピリジン（０．０９４ｍＬ，１．１７ｍｍｏｌ）、無水トリフルオロ酢酸（０．２２４ｍＬ，１．５９ｍｍｏｌ）を加え、室温で終夜撹拌した。反応液に飽和炭酸水素ナトリウム水溶液を加え、クロロホルムで３回抽出し、無水硫酸ナトリウムで乾燥した。ろ過、続いて濃縮することにより得られた残渣をシリカゲルカラムクロマトグラフィー（酢酸エチル／ｎ−ヘキサン＝２／３）により精製し、表題化合物（以下、参考例１２の化合物）を茶色アモルファスとして得た（５０４ｍｇ，収率８９％）。
^１Ｈ−ＮＭＲ（４００ＭＨｚ，ＣＤＣｌ_３）δ：１．６０（９Ｈ，ｓ），３．４５（１Ｈ，ｔ，Ｊ＝６．８Ｈｚ），３．８６−３．９８（２Ｈ，ｍ），４．１０−４．２４（２Ｈ，ｍ），５．３４（２Ｈ，ｓ），７．３６−７．５３（５Ｈ，ｍ），７．６５（１Ｈ，ｄ，Ｊ＝８．８Ｈｚ），７．８８−７．９２（２Ｈ，ｍ），８．１１−８．２２（２Ｈ，ｍ）．
ＥＳＩ−ＭＳ（ｍ／ｚ）：５３５［Ｍ＋Ｈ］^＋．（ＬＣ／ＭＳ−１）Reference Example 12 5- (Benzyloxy) -1- (chloromethyl) -7- (2,2,2-trifluoroacetamido) -1,2-dihydro-3H-benzo [e] indole-3-carvone Synthesis of tert-butyl acidate:

7-((((9H-fluoren-9-yl) methoxy) carbonyl) amino) -5- (benzyloxy) -1- (chloromethyl) -1,2-dihydro-3H-benzo [e under an argon atmosphere. ] 1 M TBAF-THF solution (1.59 mL, 1.59 mmol) was added to a THF solution (3.0 mL) of tert-butyl indole-3-carboxylate (702 mg, 1.06 mmol), and the mixture was stirred at room temperature for 3 hours. did. Water was added to the reaction solution, extracted three times with chloroform, and dried over anhydrous sodium sulfate. Dichloromethane (3.0 mL), pyridine (0.094 mL, 1.17 mmol) and trifluoroacetic anhydride (0.224 mL, 1.59 mmol) were added to the residue obtained by filtration and subsequent concentration, and the mixture was stirred at room temperature overnight. It was stirred. A saturated aqueous solution of sodium hydrogen carbonate was added to the reaction solution, extracted three times with chloroform, and dried over anhydrous sodium sulfate. The residue obtained by filtration and subsequent concentration was purified by silica gel column chromatography (ethyl acetate / n-hexane = 2/3) to obtain the title compound (hereinafter, compound of Reference Example 12) as a brown amorphous substance. (504 mg, yield 89%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 1.60 (9 H, s), 3.45 (1 H, t, J = 6.8 Hz), 3.86-3.98 (2 H, m), 4 10-4.24 (2H, m), 5.34 (2H, s), 7.36-7.53 (5H, m), 7.65 (1H, d, J = 8.8Hz), 7 .88-7.92 (2H, m), 8.11-8.22 (2H, m).
ESI-MS (m / z): 535 [M + H] ⁺ . (LC / MS-1)

参考例６の化合物の代わりに、参考例１２の化合物（２６２ｍｇ，０．４９ｍｍｏｌ）を用いて、それ以外は参考例７と同様の方法にて合成を行い、表題化合物（以下、参考例１３の化合物）を黄色固体（２１５ｍｇ，収率６９％）として得た。
^１Ｈ−ＮＭＲ（４００ＭＨｚ，ＣＤＣｌ_３）δ：３．４８（１Ｈ，ｔ，Ｊ＝６．８Ｈｚ），３．６５（１Ｈ，ｍ），３．７８（１Ｈ，ｍ），３．８７（３Ｈ，ｓ），３．９１（３Ｈ，ｓ），４．６４−４．７０（１Ｈ，ｍ），４．７４−４．８２（１Ｈ，ｍ），５．２３−５．３３（２Ｈ，ｍ），６．８７（１Ｈ，ｓ），７．０３−７．１１（２Ｈ，ｍ），７．３６−７．５５（５Ｈ，ｍ），７．７３（１Ｈ，ｄ，Ｊ＝９．２Ｈｚ），７．９６（１Ｈ，ｄｄ，Ｊ＝９．２，２．０Ｈｚ），８．１８−８．２９（３Ｈ，ｍ），９．３９（１Ｈ，ｂｒｓ）．
ＥＳＩ−ＭＳ（ｍ／ｚ）：６３８［Ｍ＋Ｈ］^＋．（ＬＣ／ＭＳ−１）Reference Example 13 N- (5- (benzyloxy) -1- (chloromethyl) -3- (5,6-dimethoxy-1H-indole-2-carbonyl) -2,3-dihydro-1H-benzo [ e] Synthesis of indol-7-yl) -2,2,2-trifluoroacetamide:

The compound of Reference Example 12 (262 mg, 0.49 mmol) was used in place of the compound of Reference Example 6, and synthesis was performed in the same manner as in Reference Example 7 except for that, and the title compound (hereinafter, referred to as Reference Example 13 Compound) was obtained as a yellow solid (215 mg, yield 69%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 3.48 (1 H, t, J = 6.8 Hz), 3.65 (1 H, m), 3.78 (1 H, m), 3.87 (3 H. , S), 3.91 (3H, s), 4.64-4.70 (1H, m), 4.74-4.82 (1H, m), 5.23-5.33 (2H, m). ), 6.87 (1H, s), 7.03-7.11 (2H, m), 7.36-7.55 (5H, m), 7.73 (1H, d, J = 9.2Hz). ), 7.96 (1H, dd, J = 9.2, 2.0 Hz), 8.18-8.29 (3H, m), 9.39 (1H, brs).
ESI-MS (m / z): 638 [M + H] ⁺ . (LC / MS-1)

アルゴン雰囲気下、参考例１３の化合物（１５ｍｇ，０．０２４ｍｍｏｌ）のＴＨＦ／メタノール／水混合溶液（１．０ｍＬ／１．０ｍＬ／１．０ｍＬ）に、水酸化リチウム一水和物（１１ｍｇ，０．２６ｍｍｏｌ）を加え、室温で終夜撹拌した。反応液に水を加え、クロロホルムで３回抽出し、無水硫酸ナトリウムで乾燥した。ろ過、続いて濃縮することにより得られた残渣をシリカゲルカラムクロマトグラフィー（酢酸エチル／ｎ−ヘキサン＝１／１）により精製し、表題化合物（以下、参考例１４の化合物）を黄色固体として得た（１０ｍｇ，収率９２％）。
^１Ｈ−ＮＭＲ（４００ＭＨｚ，ＤＭＳＯ−ｄ_６）δ：３．７８（３Ｈ，ｓ），３．７９（３Ｈ，ｓ），３．７９−３．８２（１Ｈ，ｍ），４．００（１Ｈ，ｄｄ，Ｊ＝１１．６，２．８Ｈｚ），４．１０−４．１６（１Ｈ，ｍ），４．４６−４．５３（１Ｈ，ｍ），４．６３−４．７１（１Ｈ，ｍ），５．２１（２Ｈ，ｂｒｓ），５．４０（２Ｈ，ｓ），６．９２（１Ｈ，ｓ），６．９６−７．０１（２Ｈ，ｍ），７．１２（１Ｈ，ｓ），７．２０（１Ｈ，ｄ，Ｊ＝２．４Ｈｚ），７．３２−７．４４（３Ｈ，ｍ），７．５２−７．６４（３Ｈ，ｍ），７．９９（１Ｈ，ｂｒｓ），１１．４３（１Ｈ，ｂｒｓ）．
ＥＳＩ−ＭＳ（ｍ／ｚ）：５４２［Ｍ＋Ｈ］^＋．（ＬＣ／ＭＳ−１）Reference Example 14 (7-Amino-5- (benzyloxy) -1- (chloromethyl) -1,2-dihydro-3H-benzo [e] indol-3-yl) (5,6-dimethoxy-1H -Indol-2-yl) methanone synthesis:

Lithium hydroxide monohydrate (11 mg, 0 mL) was added to a THF / methanol / water mixed solution (1.0 mL / 1.0 mL / 1.0 mL) of the compound of Reference Example 13 (15 mg, 0.024 mmol) under an argon atmosphere. .26 mmol) was added and the mixture was stirred at room temperature overnight. Water was added to the reaction solution, extracted three times with chloroform, and dried over anhydrous sodium sulfate. The residue obtained by filtration and subsequent concentration was purified by silica gel column chromatography (ethyl acetate / n-hexane = 1/1) to obtain the title compound (hereinafter, compound of Reference Example 14) as a yellow solid. (10 mg, yield 92%).
¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 3.78 (3H, s), 3.79 (3H, s), 3.79-3.82 (1H, m), 4.00 (1H). , Dd, J = 11.6, 2.8 Hz), 4.10-4.16 (1H, m), 4.46-4.53 (1H, m), 4.63-4.71 (1H, m). m), 5.21 (2H, brs), 5.40 (2H, s), 6.92 (1H, s), 6.96-7.01 (2H, m), 7.12 (1H, s). ), 7.20 (1H, d, J = 2.4 Hz), 7.32-7.44 (3H, m), 7.52-7.64 (3H, m), 7.99 (1H, brs). ), 11.43 (1H, brs).
ESI-MS (m / z): 542 [M + H] ⁺ . (LC / MS-1)

アルゴン雰囲気下、Ｎ−（ｔｅｒｔ−ブトキシカルボニル）グリシン（２３ｍｇ，０．１３ｍｍｏｌ）のジクロロメタン溶液（３．０ｍＬ）にクロロギ酸エチル（０．０１２ｍＬ，０．１３ｍｍｏｌ）、トリエチルアミン（０．０１８ｍＬ）を加え、室温で１時間撹拌した。反応液を濃縮することにより得られた残渣にジクロロメタン（３．０ｍＬ）、参考例１４の化合物（２０ｍｇ，０．０４４ｍｍｏｌ）、トリエチルアミン（０．０１８ｍＬ，０．１３ｍｍｏｌ）、ＤＭＡＰ（６ｍｇ，０．０４４ｍｍｏｌ）を加え、室温で終夜撹拌した。反応液に飽和炭酸水素ナトリウム水溶液を加え、クロロホルムで３回抽出し、無水硫酸ナトリウムで乾燥した。ろ過、続いて濃縮することにより得られた残渣を分取薄層シリカゲルクロマトグラフィー（酢酸エチル／クロロホルム＝１／４）により精製し、表題化合物（以下、参考例１５の化合物）を黄色アモルファスとして得た（１１ｍｇ，収率３５％）。
^１Ｈ−ＮＭＲ（４００ＭＨｚ，ＣＤＣｌ_３）δ：１．５１（９Ｈ，ｓ），３．３０−３．４０（１Ｈ，ｍ），３．７６−３．８８（１Ｈ，ｍ），３．９４（３Ｈ，ｓ），３．９５（３Ｈ，ｓ），３．９５−４．０７（３Ｈ，ｍ），４．６４（１Ｈ，ｔ，Ｊ＝８．０Ｈｚ），４．７５（１Ｈ，ｄｄ，Ｊ＝１１．２，２．０Ｈｚ），５．１５（２Ｈ，ｓ），５．３０−５．３６（１Ｈ，ｍ），６．９２（１Ｈ，ｓ），７．０１（１Ｈ，ｓ），７．１０（１Ｈ，ｓ），７．３５−７．４９（３Ｈ，ｍ），７．４９−７．５５（２Ｈ，ｍ），７．６０（１Ｈ，ｄ，Ｊ＝８．８Ｈｚ），７．９２（１Ｈ，ｄｄ，Ｊ＝８．８，２．０Ｈｚ），８．０２−８．１０（２Ｈ，ｍ），８．３８（１Ｈ，ｂｒｓ），９．５２（１Ｈ，ｂｒｓ）．
ＥＳＩ−ＭＳ（ｍ／ｚ）：６９９［Ｍ＋Ｈ］^＋．（ＬＣ／ＭＳ−２）(Reference Example 15) (2-((5- (benzyloxy) -1- (chloromethyl) -3- (5,6-dimethoxy-1H-indole-2-carbonyl) -2,3-dihydro-1H- Synthesis of tert-butyl benzo [e] indol-7-yl) amino) -2-oxoethyl) carbamate:

Under an argon atmosphere, ethyl chloroformate (0.012 mL, 0.13 mmol) and triethylamine (0.018 mL) were added to a dichloromethane solution (3.0 mL) of N- (tert-butoxycarbonyl) glycine (23 mg, 0.13 mmol). , Stirred at room temperature for 1 hour. Dichloromethane (3.0 mL), the compound of Reference Example 14 (20 mg, 0.044 mmol), triethylamine (0.018 mL, 0.13 mmol), and DMAP (6 mg, 0.044 mmol) were added to the residue obtained by concentrating the reaction solution. ) Was added and the mixture was stirred at room temperature overnight. A saturated aqueous solution of sodium hydrogen carbonate was added to the reaction solution, extracted three times with chloroform, and dried over anhydrous sodium sulfate. The residue obtained by filtration and subsequent concentration was purified by preparative thin-layer silica gel chromatography (ethyl acetate / chloroform = 1/4) to obtain the title compound (hereinafter, compound of Reference Example 15) as a yellow amorphous substance. (11 mg, yield 35%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 1.51 (9H, s), 3.30-3.40 (1H, m), 3.76-3.88 (1H, m), 3.94. (3H, s), 3.95 (3H, s), 3.95-4.07 (3H, m), 4.64 (1H, t, J = 8.0Hz), 4.75 (1H, dd) , J = 11.2, 2.0 Hz), 5.15 (2H, s), 5.30-5.36 (1H, m), 6.92 (1H, s), 7.01 (1H, s). ), 7.10 (1H, s), 7.35-7.49 (3H, m), 7.49-7.55 (2H, m), 7.60 (1H, d, J = 8.8Hz). ), 7.92 (1H, dd, J = 8.8, 2.0 Hz), 8.02-8.10 (2H, m), 8.38 (1H, brs), 9.52 (1H, brs). ).
ESI-MS (m / z): 699 [M + H] ⁺ . (LC / MS-2)

参考例７の化合物の代わりに、参考例１５の化合物（１１ｍｇ，０．０１５ｍｍｏｌ）を用いて、反応時間を４０分間として、それ以外は実施例１と同様の方法にて合成を行い、脱ベンジル体を白色固体として得た。得られた固体に４Ｎ塩化水素／ジオキサン溶液（１．０ｍＬ）を加え、室温で１時間撹拌した。反応液を濃縮することにより、表題化合物（以下、実施例４の化合物）を白色固体として得た（５ｍｇ，収率６６％）。
^１Ｈ−ＮＭＲ（４００ＭＨｚ，ＤＭＳＯ−ｄ_６）δ：３．４０−３．４８（１Ｈ，ｍ），３．６４−３．６９（１Ｈ，ｍ），３．７６（３Ｈ，ｓ），３．７８（３Ｈ，ｓ），３．７８−３．８３（１Ｈ，ｍ），４．０２（１Ｈ，ｄｄ，Ｊ＝１１．２，３．２Ｈｚ），４．１３−４．２０（１Ｈ，ｍ），４．５０（１Ｈ，ｄｄ，Ｊ＝１１．２，２．０Ｈｚ），４．７０−４．８０（１Ｈ，ｍ），６．９３（１Ｈ，ｓ），７．０３（１Ｈ，ｄ，Ｊ＝１．６Ｈｚ），７．１２（１Ｈ，ｓ），７．６９（１Ｈ，ｄｄ，Ｊ＝９．２，２．４Ｈｚ），７．８５（１Ｈ，ｄ，Ｊ＝９．２Ｈｚ），７．９６（１Ｈ，ｓ），８．１９（３Ｈ，ｂｒｓ），８．４１（１Ｈ，ｄ，Ｊ＝２．４Ｈｚ），１０．４２（１Ｈ，ｂｒｓ），１０．７１（１Ｈ，ｂｒｓ），１１．４４（１Ｈ，ｂｒｓ）．
ＥＳＩ−ＭＳ（ｍ／ｚ）：５０９［Ｍ＋Ｈ］^＋．（ＬＣ／ＭＳ−２）Example 4 2-Amino-N- (1- (chloromethyl) -3- (5,6-dimethoxy-1H-indole-2-carbonyl) -5-hydroxy-2,3-dihydro-1H-benzo Synthesis of [e] indol-7-yl) acetamide hydrochloride:

The compound of Reference Example 15 (11 mg, 0.015 mmol) was used instead of the compound of Reference Example 7, the reaction time was 40 minutes, and synthesis was carried out in the same manner as in Example 1 except for that, and debenzylation was performed. The body was obtained as a white solid. 4N Hydrogen chloride / dioxane solution (1.0 mL) was added to the obtained solid, and the mixture was stirred at room temperature for 1 hr. The reaction mixture was concentrated to give the title compound (hereinafter, compound of Example 4) as a white solid (5 mg, yield 66%).
¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 3.40-3.48 (1H, m), 3.64-3.69 (1H, m), 3.76 (3H, s), 3 0.78 (3H, s), 3.78-3.83 (1H, m), 4.02 (1H, dd, J = 11.2, 3.2Hz), 4.13-4.20 (1H, m), 4.50 (1H, dd, J = 11.2, 2.0Hz), 4.70-4.80 (1H, m), 6.93 (1H, s), 7.03 (1H, d, J = 1.6 Hz), 7.12 (1H, s), 7.69 (1H, dd, J = 9.2, 2.4 Hz), 7.85 (1H, d, J = 9.2 Hz) ), 7.96 (1H, s), 8.19 (3H, brs), 8.41 (1H, d, J = 2.4Hz), 10.42 (1H, brs), 10.71 (1H, brs), 1 .44 (1H, brs).
ESI-MS (m / z): 509 [M + H] ⁺ . (LC / MS-2)

アルゴン雰囲気下、参考例８の化合物（７ｍｇ，０．０１２ｍｍｏｌ）のジクロロメタン溶液（２．０ｍＬ）に（２−アミノエチル）カルバミン酸 ｔｅｒｔ−ブチル（０．００３ｍＬ，０．０１８ｍｍｏｌ）、トリエチルアミン（０．００６ｍＬ，０．０４０ｍｍｏｌ）、ＥＤＣＩ（４ｍｇ，０．０１８ｍｍｏｌ）を加え、室温で終夜撹拌した。反応液に飽和炭酸水素ナトリウム水溶液を加え、クロロホルムで３回抽出し、無水硫酸ナトリウムで乾燥した。ろ過、続いて濃縮することにより得られた残渣をシリカゲルカラムクロマトグラフィー（酢酸エチル／ｎ−ヘキサン＝２／１→１／０）により精製し、表題化合物（以下、参考例１６の化合物）を黄色アモルファスとして得た（４ｍｇ，収率５１％）。
^１Ｈ−ＮＭＲ（４００ＭＨｚ，ＣＤＣｌ_３）δ：１．４１（９Ｈ，ｓ），３．４１−３．４８（４Ｈ，ｍ），３．５７−３．６４（２Ｈ，ｍ），３．９２（３Ｈ，ｓ），３．９５（３Ｈ，ｓ），４．０７−４．１５（１Ｈ，ｍ），４．６３−４．６９（１Ｈ，ｍ），４．７５−４．８０（１Ｈ，ｍ），５．０１−５．１０（１Ｈ，ｍ），５．２８−５．３７（２Ｈ，ｍ），６．８６（１Ｈ，ｓ），７．０２（１Ｈ，ｓ），７．１０（１Ｈ，ｓ），７．３３−７．３７（１Ｈ，ｍ），７．４０−７．４６（３Ｈ，ｍ），７．５４−７．５８（２Ｈ，ｍ），７．６６−７．７３（１Ｈ，ｍ），７．９０−７．９８（１Ｈ，ｍ），８．２０（１Ｈ，ｓ），８．７７（１Ｈ，ｓ），９．３６（１Ｈ，ｓ）．
ＥＳＩ−ＭＳ（ｍ／ｚ）：７１３［Ｍ＋Ｈ］^＋．（ＬＣ／ＭＳ−２）Reference Example 16 (2- (5- (benzyloxy) -1- (chloromethyl) -3- (5,6-dimethoxy-1H-indole-2-carbonyl) -2,3-dihydro-1H-benzo [E] Synthesis of indole-7-carboxamido) ethyl) tert-butyl carbamate:

Under an argon atmosphere, tert-butyl (2-aminoethyl) carbamate (0.003 mL, 0.018 mmol) and triethylamine (0. 0) were added to a dichloromethane solution (2.0 mL) of the compound of Reference Example 8 (7 mg, 0.012 mmol). 006 mL, 0.040 mmol) and EDCI (4 mg, 0.018 mmol) were added, and the mixture was stirred at room temperature overnight. A saturated aqueous solution of sodium hydrogen carbonate was added to the reaction solution, extracted three times with chloroform, and dried over anhydrous sodium sulfate. The residue obtained by filtration and subsequent concentration is purified by silica gel column chromatography (ethyl acetate / n-hexane = 2/1 → 1/0), and the title compound (hereinafter, compound of Reference Example 16) is yellow. Obtained as an amorphous substance (4 mg, yield 51%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 1.41 (9H, s), 3.41-3.48 (4H, m), 3.57-3.64 (2H, m), 3.92. (3H, s), 3.95 (3H, s), 4.07-4.15 (1H, m), 4.63-4.69 (1H, m), 4.75-4.80 (1H). , M), 5.01-5.10 (1H, m), 5.28-5.37 (2H, m), 6.86 (1H, s), 7.02 (1H, s), 7. 10 (1H, s), 7.33-7.37 (1H, m), 7.40-7.46 (3H, m), 7.54-7.58 (2H, m), 7.66- 7.73 (1H, m), 7.90-7.98 (1H, m), 8.20 (1H, s), 8.77 (1H, s), 9.36 (1H, s).
ESI-MS (m / z): 713 [M + H] ⁺ . (LC / MS-2)

参考例７の化合物の代わりに、参考例１６の化合物（４ｍｇ，０．００６ｍｍｏｌ）を用いて、反応時間を４０分として、それ以外は実施例１と同様の方法にて合成を行い、脱ベンジル体を白色固体として得た。得られた脱ベンジル体に４Ｎ塩化水素／ジオキサン溶液（２．０ｍＬ）を加え、室温で１時間撹拌した。反応液を濃縮することにより、表題化合物（以下、参考例１７の化合物）を無色固体として得た（３ｍｇ，収率９２％）。
^１Ｈ−ＮＭＲ（４００ＭＨｚ，ＤＭＳＯ−ｄ_６）δ：３．２２−３．４８（６Ｈ，ｍ），３．５５（３Ｈ，ｓ），３．５７（３Ｈ，ｓ），３．６３−３．６７（１Ｈ，ｍ），３．７５−３．８０（１Ｈ，ｍ），３．９５−４．０４（１Ｈ，ｍ），４．３３（１Ｈ，ｄｄ，Ｊ＝１０．８，２．０Ｈｚ），４．４１（１Ｈ，ｔ，Ｊ＝５．２Ｈｚ），４．５３−４．６０（１Ｈ，ｍ），６．７３（１Ｈ，ｓ），６．８６（１Ｈ，ｄ，Ｊ＝１．６Ｈｚ），６．９１（１Ｈ，ｓ），７．４２−７．６９（１Ｈ，ｍ），７．６９−７．７４（２Ｈ，ｍ），７．８２（１Ｈ，ｓ），８．４８（１Ｈ，ｄ，Ｊ＝１．２Ｈｚ），８．５０−８．５８（１Ｈ，ｍ），１１．２７（１Ｈ，ｄ，Ｊ＝２．０Ｈｚ）．
ＥＳＩ−ＭＳ（ｍ／ｚ）：５２３［Ｍ＋Ｈ］^＋．（ＬＣ／ＭＳ−２）Reference Example 17 N- (2-aminoethyl) -1- (chloromethyl) -3- (5,6-dimethoxy-1H-indole-2-carbonyl) -5-hydroxy-2,3-dihydro-1H -Synthesis of Benzo [e] indole-7-carboxamide hydrochloride:

The compound of Reference Example 16 (4 mg, 0.006 mmol) was used in place of the compound of Reference Example 7, the reaction time was 40 minutes, and synthesis was carried out in the same manner as in Example 1 except for that, and debenzylation was performed. The body was obtained as a white solid. A 4N hydrogen chloride / dioxane solution (2.0 mL) was added to the obtained debenzylated product, and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated to give the title compound (hereinafter, compound of Reference Example 17) as a colorless solid (3 mg, yield 92%).
¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 3.22-3.48 (6H, m), 3.55 (3H, s), 3.57 (3H, s), 3.63-3. .67 (1H, m), 3.75-3.80 (1H, m), 3.95-4.04 (1H, m), 4.33 (1H, dd, J = 10.8, 2. 0 Hz), 4.41 (1H, t, J = 5.2 Hz), 4.53-4.60 (1H, m), 6.73 (1H, s), 6.86 (1H, d, J = 1.6 Hz), 6.91 (1H, s), 7.42-7.69 (1H, m), 7.69-7.74 (2H, m), 7.82 (1H, s), 8 .48 (1H, d, J = 1.2 Hz), 8.50-8.58 (1H, m), 11.27 (1H, d, J = 2.0 Hz).
ESI-MS (m / z): 523 [M + H] ⁺ . (LC / MS-2)

The analysis conditions of LC / MS-1 are as follows.
(LC / MS-1)
Liquid chromatograph system: LC1200 (Agilent Technology)
Mass spectrometer: 6130A (manufactured by Agilent Technologies)
Analytical conditions Column: Rapid resolution HT cartridge Mobile phase A: 0.1 vol% formic acid-distilled water, HPLC mobile phase B: 0.1 vol% formic acid-acetonitrile, HPLC gradient conditions: Mobile layer B vol%
0-1.5 min: 20% -95%
1.5-3.0 min: 95%
Flow rate: 0.5 mL / min
Injection volume: 2.0 μL
Column temperature: 40 ° C

The analysis conditions of LC / MS-2 are as follows.
(LC / MS-2)
Liquid chromatograph system: LC1260 Infinity II (manufactured by Agilent Technologies)
Mass spectrometer: 6130B (manufactured by Agilent Technologies)
Analytical conditions Column: Rapid resolution HT cartridge Mobile phase A: 0.1 vol% formic acid-distilled water, HPLC mobile phase B: 0.1 vol% formic acid-acetonitrile, HPLC gradient conditions: Mobile layer B vol%
0-1.5 min: 20% -95%
1.5-3.0 min: 95%
Flow rate: 0.5 mL / min
Injection volume: 2.0 μL
Column temperature: 40 ° C

(Example 5) In vitro cytotoxicity evaluation of indoline derivative (I) or a pharmacologically acceptable salt thereof:
Using the compounds of Examples 1 to 4 and the compound of Reference Example 17 as test substances, cytotoxicity against L1210 cells was measured using the MTS method.

Table 2 shows the cytotoxicity of each test substance against L1210 cells. In Table 2, EC ₅₀ means 50% effective concentration, represents the concentration showing 50% of the maximum response from the minimum value of the pharmacological action of the drug, and L1210 represents L1210 cells.

  As is clear from the results in Table 2, it was shown that the indoline derivative (I) or a pharmaceutically acceptable salt thereof has high cytotoxicity to cancer cells.

The indoline derivative (I) of the present invention or a pharmacologically acceptable salt thereof has high cytotoxicity, and thus can be used as a cytotoxic agent.

Claims (5)

An indoline derivative represented by the following general formula (I) or a pharmaceutically acceptable salt thereof.

[In the _{formula, X, -CO 2 Me, -CO 2 Et} , represent _{-C (= O) NH (CH} 2) 2 Y or ^{-NHC (= O) R 2,} Y is the one hydrogen atom It represents a phenyl group substituted with NH ₂ , R ¹ represents a hydrogen atom or a methoxy group, and R ² represents a methyl group, an ethyl group or —CH ₂ NH ₂ . ] X is -C (= O) a NH _{(CH 2)} 2 Y or ^{-NHC (= O) R 2,} R 2 is a methyl group or _-CH 2 NH _2, indoline derivative of claim 1, wherein Or a pharmaceutically acceptable salt thereof. The indoline derivative or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein R ¹ is a hydrogen atom. X is, -C (= O) NH ( CH 2) 2 Y or -NHC (= O) is _CH 2 NH _2, Y is 4-aminophenyl group, any one of claims 1 to 3 one The indoline derivative or the pharmaceutically acceptable salt thereof according to the item 1.   A cytotoxic agent comprising the indoline derivative according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof as an active ingredient.