JP2008208074A - Anticancer agent containing pyrazine derivative as active ingredient - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a pyrazine derivative having a treating effect against cancer. <P>SOLUTION: This anticancer agent contains a compound expressed by formula I [wherein, A is a specific heterocyclic group; and X is N or C] as an active ingredient. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to an anticancer agent containing a pyrazine derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

  In recent years, cancer has been the leading cause of death among Japanese people, and today, about 1 in 3 deaths occur in cancer.

  Cancer includes esophageal cancer, stomach cancer, colon cancer, liver cancer, gallbladder cancer, pancreatic cancer, breast cancer, gingival cancer, tongue cancer, head and neck cancer, ovarian cancer, uterine cancer, kidney Cancer, bladder cancer, prostate cancer, lung cancer, bone / soft tissue tumor, skin cancer, malignant melanoma, brain tumor, leukemia, malignant lymphoma, malignant pleural mesothelioma.

  Today's cancer treatment is frequently used as an effective treatment for early-stage cancer, as well as chemotherapy, hormone therapy, and radiation therapy, and for cancer that has metastasized and recurred, chemotherapy is often used as an effective treatment. Yes. Currently, cancer chemotherapy uses alkylating agents, nucleic acid antimetabolites, antitumor antibiotics, microtubule inhibitors, platinum preparations, topoisomerase inhibitors, and molecular targeted therapies. It is difficult to sufficiently treat cancer with cancer drugs, and there are currently no drugs that can be fully satisfied as anticancer drugs. In addition, these anticancer drugs are considered to have serious side effects such as digestive symptoms accompanied by nausea and vomiting, bone marrow suppression, liver dysfunction, renal dysfunction, interstitial pneumonia, cardiotoxicity, and hair loss. Yes. Therefore, development of an anticancer agent having few side effects and exhibiting a remarkable anticancer action is eagerly desired.

On the other hand, Patent Document 1 is disclosed as a pyrazine derivative having an anticancer action, but it cannot be said that the effect of inhibiting tumor growth at the individual level is sufficient. Although the pyrazine derivative disclosed in Patent Document 2 has been described as having a possibility of being applied to cancer, it has only been shown to have an inhibitory activity against a specific phosphorylase, and the cancer cell growth inhibitory action is It is not disclosed at all. In addition, the pyrazine derivative of Patent Document 3 shows a hydrogen atom, a methyl group, a halogen, a hydroxyl group, a methoxy group, a cyano group, an amino group, etc. as substituents of indole and indazole bonded to pyrazine. The amino group is not included and is different from the pyrazine derivative which is an active ingredient of the present invention. And although the pyrazine derivative of patent document 3 is disclosed as a nephritis therapeutic agent, it is not reported that the effective effect with respect to cancer is shown.
WO02 / 24681 WO03 / 93297 JP2006-045119

  It is difficult to sufficiently treat cancer with conventional anticancer agents, and serious side effects are a problem. Therefore, development of an anticancer agent having few side effects and exhibiting a remarkable anticancer action is eagerly desired.

  As a result of intensive studies, the present inventors have found that a specific pyrazine derivative or a pharmaceutically acceptable salt thereof is effective for the treatment of cancer, and have reached the present invention.

  That is, the present invention includes the following inventions.

  (1) Formula I

[Wherein, n represents an integer of 0-2, R ¹ represents a hydrogen atom or a C ₁ -C ₄ linear alkyl group, A represents an indolediyl group, a pyrrolediyl group, a furandiyl group, a thiophene group, Diyl group, pyrazole diyl group, isoxazole diyl group, isothiazole diyl group, imidazole diyl group, oxazole diyl group, thiazole diyl group or triazole diyl group, X is a nitrogen atom or CH, R ² and R ³ May be the same or different, and R ² and R ³ do not become hydrogen at the same time, C ₁ -C ₈ linear alkyl group, C ₃ -C ₈ branched alkyl group, C _3- A cyclic alkyl group of C ₈ (these alkyl groups may be substituted by a hydroxyl group, an amino group, a saturated or unsaturated 5- or 6-membered heterocyclic group), -C (O) R ⁴ , -SO It represents ₂ R ^4, R ⁴ is hydrogen, straight-chain alkyl group of C ₁ -C _3, C Branched alkyl group having ₃ -C _5, represents a phenyl group. R ² and R ³ may include a nitrogen atom to which they are bonded to form a saturated or unsaturated 5- or 6-membered heterocyclic group, and this heterocyclic group is one or more nitrogen, oxygen And may further contain a sulfur atom. In this heterocyclic group, one or two hydrogen atoms of a C ₁ -C ₄ alkyl group, a hydroxyl group, a C ₁ -C ₄ alkoxy group, an amino group, or an amino group which may be substituted with a hydroxyl group are each independently C ₁ linear alkyl group -C _3, may be substituted by an amino group which is substituted by branched alkyl groups of C ₃ -C _5. The nitrogen atom to which R ² and R ³ are bonded may be bonded to any of the 3, 4, 5, 6 and 7 positions of Formula I. ]
The anticancer agent which uses the pyrazine derivative represented by these, or its pharmaceutically acceptable salt as an active ingredient.

  (2) The anticancer agent which uses the pyrazine derivative or its pharmaceutically acceptable salt as described in said (1) whose n is 0 or 1 in Formula I as an active ingredient.

  (3) In the formula I, A is an indolediyl group, pyrrolediyl group, frangiyl group, pyrazolediyl group, and the active ingredient is the pyrazine derivative or the pharmaceutically acceptable salt thereof according to (2). Narcotic.

  (4) The anticancer agent which uses as an active ingredient the pyrazine derivative as described in said (3) whose X is a nitrogen atom in Formula I, or its pharmaceutically acceptable salt.

  (5) The pyrazine derivative or the pharmaceutically acceptable salt thereof according to any one of (1) to (4) above, wherein the cancer is colon cancer, lung cancer, prostate cancer, liver cancer, breast cancer or leukemia. Anticancer drugs.

  ADVANTAGE OF THE INVENTION According to this invention, the anticancer agent which shows sufficient therapeutic effect with respect to cancer and has few side effects can be provided.

  The pyrazine derivative which is an active ingredient of the anticancer agent of the present invention is represented by the above formula I.

  In the formula I, n represents an integer of 0-2.

R ¹ represents a hydrogen atom or a C ₁ -C ₄ linear alkyl group, preferably a hydrogen atom, a methyl group or an ethyl group, and more preferably a hydrogen atom or a methyl group.

  A represents an indole diyl group, a pyrrole diyl group, a furandiyl group, a thiophene diyl group, a pyrazole diyl group, an isoxazole diyl group, an isothiazole diyl group, an imidazole diyl group, an oxazole diyl group, a thiazole diyl group or a triazole diyl group; Preferably, it represents an indole diyl group, a pyrrole diyl group, a furandiyl group, a pyrazole diyl group, an isoxazole diyl group, an imidazole diyl group, an oxazole diyl group, or a triazole diyl group, and more preferably an indole diyl group, a pyrrole diyl group, a flange An yl group, a pyrazole diyl group, an isoxazole diyl group, or an imidazole diyl group;

The “C ₁ -C ₈ linear alkyl group” in R ² and R ³ preferably represents a C ₁ -C ₆ linear alkyl group, more preferably a C ₁ -C ₄ linear alkyl group. The “C ₃ -C ₈ branched alkyl group” preferably represents a C ₃ -C ₇ branched alkyl group, more preferably a C ₃ -C ₅ branched alkyl group. The “C ₃ -C ₈ cyclic alkyl group” preferably represents a C ₄ -C ₇ cyclic alkyl group, more preferably a C ₅ -C ₆ cyclic alkyl group. “Saturated or unsaturated 5- or 6-membered heterocyclic group” means pyrrolyl group, pyrazolyl group, imidazolyl group, thiazolyl group, oxazolyl group, isoxazolyl group, pyrrolidyl group, piperidyl group, piperidino group, piperazinyl group, piperazino group, pyridyl group Group, pyridazyl group, pyrimidyl group, morpholinyl group, morpholino group, thiomorpholinyl group, thiomorpholino group, preferably pyrrolyl group, pyrazolyl group, imidazolyl group, thiazolyl group, pyrrolidyl group, piperidyl group, piperidino group, piperazinyl group, Represents a piperazino group, a pyridyl group, a pyridazyl group, a pyrimidyl group, a morpholinyl group, a morpholino group, more preferably a pyrrolyl group, a pyrazolyl group, an imidazolyl group, a pyrrolidyl group, a piperidyl group, a piperidino group, a piperazinyl group, a piperazino group, Lysyl group, a morpholinyl group, a morpholino group.

“Substituent introduced into a saturated or unsaturated 5- or 6-membered heterocyclic group” may be a C ₁ -C ₄ alkyl group, hydroxyl group, C ₁ -C ₄ alkoxy group, amino group optionally substituted by a hydroxyl group Represents an amino group in which one or two hydrogen atoms of the amino group are each independently substituted by a C ₁ -C ₃ linear alkyl group or a C ₃ -C ₅ branched alkyl group, preferably by a hydroxyl group An optionally substituted C ₁ -C ₄ alkyl group, a C ₁ -C ₄ alkoxy group, one or two hydrogen atoms of the amino group are each independently a C ₁ -C ₃ linear alkyl group, C ₃ -C ₅ Represents an amino group substituted by a branched alkyl group, and more preferably, a C ₁ -C ₄ alkoxy group, a linear alkyl group in which one or two hydrogen atoms of the amino group are each independently C ₁ -C ₃ , an amino group which is substituted by branched alkyl groups of C ₃ -C _5.

R ⁴ represents a hydrogen atom, a C ₁ -C ₃ linear alkyl group, a C ₃ -C ₅ branched alkyl group, or a phenyl group, preferably a hydrogen atom, a C ₁ -C ₃ linear alkyl group, C _It represents a ₃ -C ₅ branched alkyl group, more preferably a C ₁ -C ₃ linear alkyl group, or a C ₃ -C ₅ branched alkyl group.

  Preferable specific examples of the compound represented by the formula I include compounds described in the following Tables 1 to 14, pharmaceutically acceptable salts thereof, and reference examples described in Examples. However, the present invention is not limited to them.

  The production method of the compound represented by Formula I (hereinafter, for example, “the compound represented by Formula I” may be simply represented as “I”) is shown below. It is not limited to. In various production methods, reaction conditions are appropriately selected from those described below.

Among the compounds represented by formula I, II in which R ¹ is hydrogen is a mixture of I in an alcoholic solvent such as methanol, tetrahydrofuran, dimethoxyethane, 1,4-dioxane, sodium hydroxide aqueous solution, hydroxide It can be produced by hydrolysis with a base such as an aqueous lithium solution or an aqueous barium hydroxide solution. Although reaction temperature is not specifically limited, Usually, it is about 0 degreeC-room temperature, and reaction time is about 1 hour-48 hours. The amount of the base added is usually about 1 to 5 equivalents relative to I.

(Wherein A, X and R ² and R ³ are the same as defined above).

  I can be produced by the following steps (in the present specification, “step” is described as “step” during the chemical reaction).

(Wherein R ⁵ represents chlorine, bromine or iodine, and A, X and R ² and R ³ are the same as defined above).

  When the 2-position of the pyrazine ring is bonded to the carbon atom of A, Step 1 can be performed by the method described in “Palladium in Heterocyclic Chemistry vol. 20, 2000, PERGAMON” and its cited references. That is, in a solvent such as dimethylformamide, dioxane, tetrahydrofuran, dimethoxyethane, III and V in the presence of 3 mol% to 20 mol% of a palladium catalyst with respect to III and 1 to 4 equivalents of a tertiary amine or inorganic base with respect to III. Can be carried out by reacting. Palladium catalysts include tetrakis (triphenylphosphine) palladium, palladium acetate, [1,1'-bis (diphenylphosphino) ferrocene] palladium (II) dichloride dichloromethane complex, bis (methyldiphenylphosphine) palladium (II) dichloride, bis (2,4-pentanedionate) palladium (II), bis (triphenylphosphine) palladium (II) dichloride, and the like are used, but not limited thereto. Further, triphenylphosphine, tri-o-tolylphosphine, tetrabutylammonium chloride and the like may be added as appropriate. Examples of the tertiary amine include, but are not limited to, triethylamine and diisopropylethylamine. Examples of the inorganic base include potassium carbonate, potassium phosphate, sodium carbonate, cesium carbonate, but are not limited thereto. Although reaction temperature is not specifically limited, It is about 60 to 140 degreeC, and reaction time is about 2 to 12 hours.

Further, it can also be produced by “Palladium in Heterocyclic Chemistry vol. 20, 2000, PERGAMON” and the following method (Suzuki coupling) described in the cited reference. That is, in a solvent such as dimethylformamide, dioxane, tetrahydrofuran, dimethoxyethane, III and R in the presence of 3 mol% to 20 mol% of a palladium catalyst with respect to III and 1 to 4 equivalents of a tertiary amine or inorganic base with respect to III. ¹ OOC— (CH ₂ ) _n —AB (OH) ₂ (R ₁ and A are as defined above) can be produced by reacting. Palladium catalysts include tetrakis (triphenylphosphine) palladium, palladium acetate, [1,1'-bis (diphenylphosphino) ferrocene] palladium (II) dichloride dichloromethane complex, bis (methyldiphenylphosphine) palladium (II) dichloride, bis (2,4-pentanedionate) palladium (II), bis (triphenylphosphine) palladium (II) dichloride, and the like are used, but not limited thereto. Examples of tertiary amines and inorganic bases include triethylamine, diisopropylethylamine, potassium carbonate, potassium phosphate, sodium carbonate, and cesium carbonate, but are not limited thereto. Although reaction temperature is not specifically limited, Usually, it is about 60 to 140 degreeC, and reaction time is about 2 to 12 hours.

When the 2-position of the pyrazine ring is bonded to the nitrogen atom of A, step 1 can be performed by the method described in “Angew. Chem. Int. Ed. 2003, 42, 5400-5449” and the cited references. . That is, it can be carried out by reacting III and V in the presence of a catalytic amount of copper, organic amine and base in a solvent such as toluene, benzene, dimethylformamide, dimethyl sulfoxide, tetrahydrofuran or dioxane. Examples of copper include, but are not limited to, Cu, CuI, CuCl, CuSCN, Cu ₂ O, CuCl ₂ , CuSO ₄ , Cu (OAc) ₂ , and Cu (acac) ₂ . Organic amines include ethylenediamine, N, N'-dimethylethylenediamine, N-methyl-N'-dimethylethylenediamine, N, N-dimethylethylenediamine, Nn-butylethylenediamine, N-methylethylenediamine, cyclohexane 1,2-diamine, N, N′-dimethylcyclohexane-1,2-diamine and the like are used, but not limited thereto. Further, as the base, potassium phosphate, potassium carbonate, cesium carbonate, sodium t-butoxide and the like are used, but not limited thereto. The amount of copper used is about 5 mol% to 20 mol% with respect to III, and the amount of organic amine is about 5 mol% to 40 mol% with respect to III. The base is used in an amount of about 1 to 4 equivalents with respect to III. Although reaction temperature is not specifically limited, Usually, it is about 70 to 140 degreeC, and reaction time is about 12 to 48 hours.

  Step 2 can be performed in the same manner as in step 1.

  In V, XI in which A is pyrrole, n is 1, and the substitution position is the 3-position can be produced by the following steps.

(Wherein R ₁ is the same as defined above).

  Step 1 is a step of protecting the nitrogen atom of pyrrole with a triisopropylsilyl group (TIPS). In a solvent such as dimethylformamide, tetrahydrofuran, or dimethoxyethane, VII is reacted with a base such as sodium hydride or potassium t-butoxide with VII, and then 1 equivalent of triisopropylsilyl chloride with VII. It can be carried out by reacting. The amount of the base and triisopropylsilyl chloride can be appropriately increased. The reaction temperature is not particularly limited, but is about 0 ° C. to 50 ° C., and the reaction time is about 1 hour to 5 hours. In addition to triisopropylsilyl chloride, t-butyldiphenylsilyl chloride can also be used to protect with a bulky protecting group.

  In step 2, in a solvent such as dichloromethane or chloroform, about 1 to 3 equivalents of alkyloxyoxalyl chloride and VIII are added in the presence of about 1 to 3 equivalents of a tertiary amine such as pyridine, triethylamine, or diisopropylethylamine. It can be carried out by reacting. The reaction temperature is not particularly limited, but is about -20 ° C to 50 ° C, and the reaction time is about 2 hours to 48 hours.

  Step 3 is a step of deprotecting the triisopropylsilyl group. It can be carried out by reacting IX with 1 to 3 equivalents of tetra-n-butylammonium fluoride with respect to IX in a solvent such as tetrahydrofuran or dimethoxyethane. The reaction temperature is not particularly limited, but is about 0 ° C. to 50 ° C., and the reaction time is about 1 hour to 5 hours.

  In step 4, in the presence of a catalytic amount of an acid catalyst such as p-toluenesulfonic acid in an alcoholic solvent such as methanol or ethanol, 1 to 3 equivalents of p-toluenesulfonylhydrazide and X are reacted. It can be carried out by reduction with 1 to 3 equivalents of sodium cyanoborohydride with respect to X under acidic conditions. In order to make it acidic, dilute hydrochloric acid or dilute sulfuric acid is usually used, but it is not limited thereto. The reaction temperature between p-toluenesulfonyl hydrazide and X is not particularly limited, but is about 50 ° C. to 100 ° C., and the reaction time is about 1 to 6 hours. The reduction reaction temperature with sodium cyanoborohydride is not particularly limited, but is about room temperature to about 100 ° C., and the reaction time is about 2 hours to 6 hours.

In VI, XV in which X is a nitrogen atom and has a substituent —NR ² R ³ can be produced by the following steps.

(Wherein R ² , R ³ and R ⁵ are the same as defined above).

Step 1 is a step of coupling indazole having a substituent R ^{5 in} which indazole 1-position is protected with a tetrahydropyranyl group (THP) and amine HNR ² R ³ .

  That is, in a solvent such as dimethylformamide, dioxane, tetrahydrofuran, dimethoxyethane, toluene, benzene, 3 mol% to 20 mol% of palladium catalyst with respect to XII and 1 to 4 equivalents of tertiary amine, inorganic base or organic with respect to XII The reaction can be carried out by reacting XII with 1 to 4 equivalents of XIII in the presence of a base. Palladium catalysts include tetrakis (triphenylphosphine) palladium, palladium acetate, tris (dibenzylideneacetone) dipalladium, [1,1'-bis (diphenylphosphino) ferrocene] palladium (II) dichloride dichloromethane complex, bis (methyldiphenyl) Phosphine) palladium (II) dichloride, bis (2,4-pentanedionate) palladium (II), bis (triphenylphosphine) palladium (II) dichloride and the like are used, but not limited thereto. Further, triphenylphosphine, tri-o-tolylphosphine, BINAP, tetrabutylammonium chloride and the like may be added as appropriate. Examples of the tertiary amine include, but are not limited to, triethylamine and diisopropylethylamine. Examples of the inorganic base include potassium carbonate, potassium phosphate, sodium carbonate, cesium carbonate, and examples of the organic base include sodium methoxide, sodium tert-butoxide, and the like, but are not limited thereto. Although reaction temperature is not specifically limited, It is about 60 to 140 degreeC, and reaction time is about 2 to 12 hours.

  Step 2 is a step of deprotecting the tetrahydropyranyl group. It can be carried out by reacting 1 to 3 equivalents of an aqueous hydrochloric acid solution with respect to XIV in a solvent such as tetrahydrofuran, dimethoxyethane, dioxane and the like. In order to make it acidic, dilute hydrochloric acid or dilute sulfuric acid is usually used, but it is not limited thereto. The reaction temperature is not particularly limited, but is about 0 ° C. to 80 ° C., and the reaction time is about 2 hours to 12 hours.

XVIII having X as a nitrogen atom and a substituent —NHR ² can be produced by the following steps.

(Wherein R ¹ and A are the same as defined above).

Step 1 is an aldehyde having 1 to 3 equivalents of the substituent R ⁶ relative to XVI or a ketone XVII having the substituents R ⁷ and R ⁸ under acidic conditions in a solvent such as methanol, ethanol, dioxane, dimethylformamide and the like. After reacting XVI, the resulting imine can be reduced by 1-3 equivalents of sodium cyanoborohydride, sodium borohydride, sodium triacetoxyborohydride or solid phase supported sodium borohydride . In order to make it acidic, acetic acid, dilute hydrochloric acid, dilute sulfuric acid and the like are usually used, but not limited thereto. The reaction temperature is not particularly limited, but is about room temperature to 80 ° C., and the reaction time is about 2 hours to 24 hours.

  The reaction products obtained by the respective production methods are isolated and purified as various solvates such as a free compound, a salt or a hydrate thereof. The salt can be produced by subjecting it to a normal salt formation treatment. Isolation and purification are performed by adapting ordinary operations such as extraction, concentration, distillation, crystallization, filtration, recrystallization, and various chromatography.

  Examples of the pharmaceutically acceptable salt of the compound represented by the formula I include ammonium salts, alkali metal salts (for example, sodium salts and potassium salts), alkaline earth metal salts (for example, calcium salts, and magnesium salts). ) Or other organic bases such as dicyclohexylamine, N-methyl-D-glucamine, ethanolamine, diethanolamine, triethanolamine, diisopropanolamine, tris (hydroxymethyl) aminomethane Amino acid salts such as salt, lysine salt and arginine salt, hydrohalide such as hydrofluoride, hydrochloride, hydrobromide, hydroiodide, nitrate, perchlorate, sulfate Inorganic acids such as phosphate, methanesulfonate, trifluoromethanesulfonate, ethanesulfonate Aryl sulfonates such as alkyl sulfonate, benzene sulfonate, p-toluene sulfonate, fumarate, succinate, citrate, tartrate, oxalate, maleate, acetate, Examples thereof include organic acid salts such as malate, lactate, and ascorbate.

  Effective cancers of the anticancer agent of the present invention include esophageal cancer, stomach cancer, colon cancer, liver cancer, gallbladder cancer, pancreatic cancer, breast cancer, gingival cancer, tongue cancer, and head and neck. Cancer, ovarian cancer, uterine cancer, kidney cancer, bladder cancer, prostate cancer, lung cancer, bone / soft tissue tumor, skin cancer, malignant melanoma, brain tumor, leukemia, malignant lymphoma, malignant pleural mesothelioma, etc. And various types of cancers, including but not limited to, colon cancer, lung cancer, prostate cancer, liver cancer, breast cancer or leukemia, and more preferably colon cancer.

  The anticancer effect of the anticancer agent of the present invention can be evaluated by a growth inhibitory action against various cultured cancer cell lines, and a tumor growth inhibitory action or a survival days extending action in various cancer animals. it can. However, the evaluation method is not limited to these.

  The compound represented by the above formula I, which is an active ingredient of the anticancer agent of the present invention, and a pharmaceutically acceptable salt thereof are used as mammals (for example, as a powder or as a pharmaceutical composition in an appropriate dosage form). , Mice, rats, hamsters, rabbits, dogs, monkeys, cows, sheep, humans, etc.) or orally (eg, transdermal administration, intravenous administration, rectal administration, inhalation administration, nasal administration, ophthalmic administration) Administration).

  Specific examples of the dosage form for administration include tablets, powders, pills, capsules, granules, syrups, solutions, injections, emulsions, suspensions, suppositories and the like. Such dosage forms are produced by a method known per se and contain various carriers usually used in the pharmaceutical field. Examples include excipients, lubricants, binders, disintegrants in solid preparations; solvents, solubilizers, suspending agents, soothing agents, etc. in liquid preparations. If necessary, additives such as preservatives, antioxidants, colorants, sweeteners, adsorbents, wetting agents and the like can also be used. Also included are known sustained-release preparations.

  Examples of the excipient include lactose, D-mannitol, starch, sucrose, corn starch, crystalline cellulose, and light anhydrous silicic acid. Examples of the lubricant include magnesium stearate, calcium stearate, talc, colloidal silica and the like.

  Examples of the binder include crystalline cellulose, D-mannitol, dextrin, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, starch, sucrose, gelatin, methylcellulose, sodium carboxymethylcellulose, and the like.

  Examples of the disintegrant include starch, carboxymethyl cellulose, carboxymethyl cellulose calcium, croscarmellose sodium, carboxymethyl starch sodium, L-hydroxypropyl cellulose, and the like. Examples of the solvent include water for injection, alcohol, propylene glycol, macrogol, sesame oil, corn oil and the like.

  Examples of the solubilizer include polyethylene glycol, propylene glycol, D-mannitol, benzyl benzoate, ethanol, cholesterol, triethanolamine, sodium carbonate, sodium citrate and the like.

  Examples of the suspending agent include surfactants such as stearyltriethanolamine, sodium lauryl sulfate, laurylaminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, and glyceryl monostearate, or polyvinyl alcohol, polyvinylpyrrolidone, and methylcellulose. , Hydrophilic polymers such as hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, and the like.

  Examples of isotonic agents include glucose, sodium chloride, D-sorbitol, D-mannitol and the like.

  Examples of the buffer include buffer solutions of phosphate, acetate, carbonate, citrate and the like. Examples of soothing agents include benzyl alcohol.

  Examples of the preservative include p-hydroxybenzoates, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, sorbic acid and the like.

  Examples of the antioxidant include sulfite and ascorbic acid.

  The anticancer agent of the present invention desirably contains the above components in an amount of 0.001 to 99% by weight, more preferably 0.01 to 99% by weight. The effective dose and frequency of administration of the compound represented by Formula I, which is the active ingredient of the present invention, or a pharmaceutically acceptable salt thereof are the dosage form, patient age, body weight, nature or severity of symptoms to be treated. Although it varies depending on the dose, it is usually possible to administer 1 to 1000 mg, preferably 1 to 300 mg per day for an adult, in one or several divided doses.

  The anticancer agent of the present invention can also be used in combination with or in combination with other drugs in order to supplement or enhance the therapeutic effect of the anticancer agent of the present invention or to reduce the dose. It can be used in combination with drugs (hereinafter abbreviated as concomitant drugs) such as therapeutic agents, immunotherapeutic agents, cachexia improving drugs and the like. At this time, the administration time of the anticancer agent and the concomitant drug of the present invention is not limited, and these may be administered simultaneously to the administration subject, or may be administered with a time difference. The concomitant drug may be a low molecular compound, a high molecular protein, a polypeptide, an antibody, or a vaccine. The dose of the concomitant drug can be appropriately selected based on the clinically used dose.

  The mixing ratio of the anticancer agent of the present invention and the concomitant drug can be appropriately selected depending on the administration subject, administration route, target disease, symptom, combination and the like. For example, when the administration subject is a human, a concomitant drug may be used in an amount of 0.01 to 100 for the active ingredient 1 of the anticancer agent of the present invention.

  As chemotherapeutic agents, alkylating agents such as melphalan, cyclophosphamide, ifosfamide, busulfan, nimustine, ranimustine, temozolomide, methotrexate, fluorouracil, tegafur, carmofur, doxyfluridine, capecitabine, cytarabine, ancitabine, enocitabine, Nucleic acid metabolism antagonists such as Pharto, gemcitabine, mercaptopurine, fludarabine, doxorubicin, daunorubicin, pirarubicin, epirubicin, idarubicin, mitoxantrone, mitomycin C, bleomycin, pepromycin and other antitumor antibiotics, vincristine, vinblastine, vindesine, vinorelbine , Microtubule inhibitors such as paclitaxel and docetaxel, cisplatin, carbop Examples include, but are not limited to, platinum preparations such as latin and nedaplatin, topoisomerase inhibitors such as irinotecan, nogitecan, and etoposide, and molecular targeted therapeutics such as trastuzumab, rituximab, and imatinib.

  Examples of immunotherapeutic agents include muramyl dipeptide derivatives, lentinan, schizophyllan, ubenimex, pisibanil, krestin, interferon, interleukin (IL), granulocyte colony stimulating factor, erythropoietin and the like.

  As cachexia-improving drugs, cyclooxygenase inhibitors such as indomethacin and diclofenac, progesterone derivatives such as megesterol acetate, carbohydrate steroids such as dexamethasone, metoclopramide drugs, tetrahydrocannabinol drugs, and fat metabolism improvement such as eicosapentaenoic acid And antibodies against growth agents, growth hormones, IGF-1, or cachexia-inducing factors TNF-α, LIF, IL-6, and oncostatin M.

  EXAMPLES The present invention will be specifically described below with reference to examples, but the present invention is not limited to the following examples.

Reference example 1
Ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate

Under argon atmosphere, trans-N, N'-dimethyl-1,2-cyclohexanediamine 780 mg, copper iodide 260 mg, ethyl 2-pyrrol-3-yl acetate 4.2 g, tripotassium phosphate n-hydrate 15.3 g, 10 g of 2,6-diiodopyrazine was suspended in 30 mL of toluene and stirred at 110 ° C. for 24 hours. The reaction mixture was filtered through silica gel (ethyl acetate) and concentrated. The residue was purified by silica gel column chromatography (n-hexane / ethyl acetate = 10/2) to obtain 5.9 g of ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate. (Yield 49%).
MS (ESI): 358 (M + H) ^+
1 H-NMR (300 MHz, CDCl ₃ ) δ: 1.28 (3H, t, J = 7.1), 3.53 (2H, s), 4.18 (2H, q, J = 7.1), 6.37 (1H, s), 7.40- 7.46 (2H, m), 8.34 (1H, s), 8.57 (1H, s).

Reference example 2
Ethyl 2- [1- (6- {6- [isopropyl (methyl) amino] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetate

Under argon atmosphere, trans-N, N'-dimethyl-1,2-cyclohexanediamine 10 mg, copper iodide 4 mg, N-isopropyl-N-methyl-1H-indazol-6-amine 76 mg, tripotassium phosphate / water 162 mg of the Japanese product and 130 mg of ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate were suspended in 1 mL of toluene and stirred at 110 ° C. for 24 hours. The reaction mixture was filtered through silica gel (ethyl acetate) and concentrated. The residue was purified by silica gel column chromatography (n-hexane / ethyl acetate = 1/1) and 139 mg of ethyl 2- [1- (6- {6- [isopropyl (methyl) amino] -1H-indazole-1- Yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetate was obtained (yield 91%).
MS (ESI): 419 (M + H) ^+
1 H-NMR (400 MHz, CDCl ₃ ) δ: 1.28 (6H, d, J = 6.6), 1.31 (3H, t, J = 7.1), 2.93 (3H, s), 3.58 (2H, s), 4.21 ( 2H, q, J = 7.1), 4.28-4.33 (1H, m), 6.43 (1H, brs), 6.95 (1H, dd, J = 2.2,9.1), 7.57-7.62 (3H, m), 8.05 (1H , brs), 8.07 (1H, s), 8.47 (1H, s), 9.17 (1H, s).

Reference example 3
2- [1- (6- {6- [Isopropyl (methyl) amino] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetic acid

Ethyl 2- [1- (6- {6- [isopropyl (methyl) amino] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetate 40 mg It melt | dissolved in dioxane 2mL, 1M sodium hydroxide aqueous solution 0.5mL was added, and it stirred at room temperature for 2 hours. The reaction solution was adjusted to neutrality by adding 1M hydrochloric acid, and extracted with ethyl acetate. The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and 26 mg of 2- [1- (6- {6- [isopropyl (methyl) amino] -1H-indazole-1- Yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetic acid was obtained (yield 67%).
MS (ESI): 391 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 1.25 (6H, d, J = 6.3), 2.91 (3H, s), 3.50 (2H, s), 4.28-4.33 (1H, m), 6.42 ( 1H, brs), 7.05 (1H, d, J = 9.3), 7.69-7.72 (2H, m), 7.79 (1H, brs), 7.95 (1H, s), 8.32 (1H, s), 8.89 (1H, s), 9.06 (1H, s).

Reference example 4
Ethyl 2- (1- {6- [6- (pyrrolidin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate

Under argon atmosphere, ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate 39 mg, copper iodide 1.0 mg, tripotassium phosphate n-hydrate 62 mg, trans-N, N′-dimethyl-1,2-cyclohexanediamine 3.2 mg and 6- (pyrrolidin-1-yl) -1H-indazole 25 mg were suspended in 0.89 mL of 1,4-dioxane and stirred at 110 ° C. for 24 hours. The reaction mixture was filtered through celite bed (THF), and the solvent was distilled off under reduced pressure. The residue obtained was purified by column chromatography (NH silica, ethyl acetate / cyclohexane = 1/4) to give ethyl 2- (1- {6- [6- (pyrrolidin-1-yl) -1H-indazole]. 20 mg of 1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate was obtained as a yellow solid (42% yield).
MS (ESI): 417 (M + H) ^+
1 H-NMR (400 MHz, CDCl ₃ ) δ: 9.12 (1H, s), 8.42 (1H, s), 8.04 (1H, d, J = 0.7), 7.75 (1H, s), 7.56-7.54 (3H, m), 6.67 (1H, dd, J = 8.8,2.2), 6.41 (1H, dd, J = 2.9,1.7), 4.19 (2H, q, J = 7.1), 3.55 (2H, s), 3.46-3.43 (4H, m), 2.11-2.08 (4H, m), 1.29 (3H, t, J = 7.1).

Reference Example 5
2- (1- {6- [6- (Pyrrolidin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetic acid

Ethyl 2- (1- {6- [6- (pyrrolidin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrole- dissolved in 1.5 mL of 1,4-dioxane To 19 mg of 3-yl) acetate, 0.45 mL of 1N aqueous sodium hydroxide solution was added, and the mixture was stirred at room temperature for 17 hours. 1N hydrochloric acid was added to the reaction solution. The precipitated precipitate was collected by filtration and washed with water to give 2- (1- {6- [6- (pyrrolidin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H 18 mg of (pyrrol-3-yl) acetic acid was obtained as a pale yellow solid (quantitative).
MS (ESI): 389 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 12.29 (1H, brs), 8.99 (1H, s), 8.83 (1H, s), 8.26 (1H, s), 7.75 (1H, dd, J = 2.6, 2.6), 7.72 (1H, brs), 7.67-7.65 (2H, m), 6.75 (1H, dd, J = 8.9, 1.8), 6.38 (1H, s), 3.46 (2H, s), 3.39 ( 4H, t, J = 6.3), 2.03 (4H, t, J = 6.3).

Reference Example 6
Ethyl 2- (1- {6- [6- (dimethylamino) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate

Under argon atmosphere, ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate 37 mg, copper iodide 1.0 mg, tripotassium phosphate n-hydrate 58 mg, trans-N , N′-dimethyl-1,2-cyclohexanediamine 2.9 mg and N, N-dimethyl-1H-indazol-6-amine 20 mg were suspended in 0.83 mL of 1,4-dioxane and stirred at 110 ° C. for 24 hours. The reaction mixture was filtered through celite bed (THF), and the solvent was distilled off under reduced pressure. The obtained residue was purified by column chromatography (NH silica, ethyl acetate / cyclohexane = 1/4) to give ethyl 2- (1- {6- [6- (dimethylamino) -1H-indazole-1- 22 mg of (yl) pyrazin-2-yl} -1H-pyrrol-3-yl) acetate was obtained as a yellow solid (55% yield).
MS (ESI): 389 (M + H) ^+
1 H-NMR (400 MHz, CDCl ₃ ) δ: 9.14 (1H, s), 8.44 (1H, s), 8.06 (1H, s), 7.93 (1H, d, J = 2.0), 7.59-7.55 (3H, m), 6.85 (1H, dd, J = 8.7, 2.2), 6.41-6.40 (1H, m), 4.19 (2H, q, J = 7.1), 3.55 (2H, s), 3.14 (6H, s), 1.29 (3H, t, J = 7.1).

Reference Example 7
2- (1- {6- [6- (Dimethylamino) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetic acid

Ethyl 2- (1- {6- [6- (dimethylamino) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl dissolved in 1.5 mL of 1,4-dioxane ) 0.56 mL of 1N aqueous sodium hydroxide solution was added to 22 mg of acetate, and the mixture was stirred at room temperature for 17 hours. 1N hydrochloric acid was added to the reaction solution. The precipitated precipitate was collected by filtration and washed with water to give 2- (1- {6- [6- (dimethylamino) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrole- 17 mg of 3-yl) acetic acid was obtained as a pale yellow solid (yield 86%).
MS (ESI): 362 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 12.29 (1H, brs), 9.02 (1H, s), 8.86 (1H, s), 8.30 (1H, s), 7.82 (1H, d, J = 1.7), 7.76 (1H, dd, J = 2.9,2.4), 7.72 (1H, brs), 7.69 (1H, d, J = 8.8), 6.95 (1H, dd, J = 8.8,2.4), 6.38-6.37 (1H, m), 3.46 (2H, s), 3.10 (6H, s).

Reference Example 8
Ethyl 2- (1- {6- [6- (piperidin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate

Under argon atmosphere, ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate 74 mg, copper iodide 2.0 mg, tripotassium phosphate n-hydrate 116 mg, trans-N , N′-dimethyl-1,2-cyclohexanediamine 5.8 mg and 6- (piperidin-1-yl) -1H-indazole 50 mg were suspended in 1.65 mL of 1,4-dioxane and stirred at 110 ° C. for 24 hours. The reaction mixture was filtered through celite bed (THF), and the solvent was distilled off under reduced pressure. The residue obtained was purified by column chromatography (NH silica, ethyl acetate / cyclohexane = 1/5) to give ethyl 2- (1- {6- [6- (piperidin-1-yl) -1H-indazole]. 39 mg of (-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate was obtained as a yellow solid (45% yield).
MS (ESI): 431 (M + H) ^+
1 H-NMR (400 MHz, CDCl ₃ ) δ: 9.14 (1H, s), 8.46 (1H, s), 8.19 (1H, s), 8.07 (1H, s), 7.60-7.54 (3H, m), 7.05 (1H, dd, J = 8.8,2.1), 6.43-6.42 (1H, m), 4.20 (2H, q, J = 7.1), 3.56 (2H, s), 3.39-3.36 (4H, m), 1.88- 1.66 (6H, m), 1.29 (3H, t, J = 7.1).

Reference Example 9
2- (1- {6- [6- (Piperidin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetic acid

Ethyl 2- (1- {6- [6- (piperidin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H- dissolved in 1,4-dioxane (1.5 mL) A 1N aqueous sodium hydroxide solution (0.91 mL) was added to 39 mg of pyrrol-3-yl) acetate, and the mixture was stirred at room temperature for 17 hours. 1N hydrochloric acid was added to the reaction solution. The precipitated precipitate was collected by filtration and washed with water to give 2- (1- {6- [6- (piperidin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H 32 mg of (pyrrol-3-yl) acetic acid was obtained as a pale yellow solid (89% yield).
MS (ESI): 402 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 12.28 (1H, brs), 9.02 (1H, s), 8.87 (1H, s), 8.32 (1H, s), 8.08 (1H, brs), 7.76 (1H, dd, J = 2.7,2.7), 7.71-7.69 (2H, m), 7.16 (1H, dd, J = 9.1,2.2), 6.40-6.38 (1H, m), 3.46 (2H, s), 3.38-3.35 (4H, m), 1.69-1.61 (6H, m).

Reference Example 10
Ethyl 2- {1- [6- (6-morpholino-1H-indazol-1-yl) pyrazin-2-yl] -1H-pyrrol-3-yl} acetate

Under argon atmosphere, ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate 71 mg, copper iodide 1.9 mg, tripotassium phosphate n-hydrate 111 mg, trans-N, N′-dimethyl-1,2-cyclohexanediamine (5.7 mg) and 6-morpholino-1H-indazole (49 mg) were suspended in 1.65 mL of 1,4-dioxane and stirred at 110 ° C. for 24 hours. The reaction mixture was filtered through celite bed (THF), and the solvent was distilled off under reduced pressure. By purifying the obtained residue by column chromatography (NH silica, ethyl acetate / cyclohexane = 1/5), ethyl 2- {1- [6- (6-morpholino-1H-indazol-1-yl) pyrazine 54 mg of -2-yl] -1H-pyrrol-3-yl} acetate was obtained as a pale yellow solid (63% yield).
MS (ESI): 433 (M + H) ^+
1 H-NMR (400 MHz, CDCl ₃ ) δ: 9.17 (1H, s), 8.49 (1H, s), 8.22 (1H, s), 8.13 (1H, s), 7.66 (1H, d, J = 8.8) , 7.58 (1H, brs), 7.53-7.52 (1H, m), 7.05 (1H, dd, J = 8.8, 2.2), 6.43-6.41 (1H, m), 4.20 (2H, q, J = 7.1), 3.95-3.93 (4H, m), 3.56 (2H, s), 3.38-3.35 (4H, m), 1.30 (3H, t, J = 7.1).

Reference Example 11
2- {1- [6- (6-morpholino-1H-indazol-1-yl) pyrazin-2-yl] -1H-pyrrol-3-yl} acetic acid

52 mg of ethyl 2- {1- [6- (6-morpholino-1H-indazol-1-yl) pyrazin-2-yl] -1H-pyrrol-3-yl} acetate dissolved in 1.5 mL of 1,4-dioxane To the mixture, 1.2 mL of 1N aqueous sodium hydroxide solution was added and stirred at room temperature for 17 hours. 1N hydrochloric acid was added to the reaction solution. The precipitated precipitate was collected by filtration and washed with water to give 2- {1- [6- (6-morpholino-1H-indazol-1-yl) pyrazin-2-yl] -1H-pyrrol-3-yl } Obtained 48 mg acetic acid as a cream colored solid (quantitative).
MS (ESI): 404 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 12.31 (1H, brs), 9.03 (1H, s), 8.89 (1H, s), 8.36 (1H, s), 8.07 (1H, brs), 7.76 -7.72 (3H, m), 7.20 (1H, dd, J = 8.8,2.0), 6.40-6.39 (1H, m), 3.83 (4H, t, J = 4.7), 3.48 (2H, s), 3.33- 3.30 (4H, m).

Reference Example 12
Ethyl 2- (1- {6- [6- (4-methylpiperidin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate

Under an argon atmosphere, ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate 101 mg, copper iodide 2.7 mg, tripotassium phosphate n-hydrate 158 mg, trans-N, N'-dimethyl-1,2-cyclohexanediamine 8.0 mg and 6- (4-methylpiperidin-1-yl) -1H-indazole 73 mg were suspended in 1.26 mL of 1,4-dioxane, and 110 ° C. for 24 hours. Stir. The reaction mixture was filtered through celite bed (THF), and the solvent was distilled off under reduced pressure. The obtained residue was purified by column chromatography (silica gel, ethyl acetate / cyclohexane = 1/5) to give ethyl 2- (1- {6- [6- (4-methylpiperidin-1-yl) -1H 99 mg of -indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate was obtained as a yellow solid (yield 80%).
MS (ESI): 444 (M + H) ^+
1 H-NMR (400 MHz, CDCl ₃ ) δ: 9.14 (1H, s), 8.46 (1H, s), 8.17 (1H, d, J = 2.0), 8.07 (1H, s), 7.59-7.54 (3H, m), 7.06-7.03 (1H, m), 6.43-6.40 (1H, m), 4.20 (2H, q, J = 7.1), 3.89-3.86 (2H, m), 3.57 (2H, s), 2.93 2.87 (2H, m), 1.81-1.79 (2H, m), 1.64-1.58 (1H, m), 1.47-1.37 (2H, m), 1.30 (3H, t, J = 7.1), 1.00 (3H, d , J = 5.6).

Reference Example 13
2- (1- {6- [6- (4-Methylpiperidin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetic acid

Ethyl 2- (1- {6- [6- (4-methylpiperidin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H dissolved in 4.0 mL of 1,4-dioxane -Pyrrol-3-yl) acetate (98 mg) was added with 1N aqueous sodium hydroxide (0.44 mL) and stirred at room temperature for 48 hours. 1N hydrochloric acid was added to the reaction solution. The precipitated precipitate was collected by filtration and washed with water to give 2- (1- {6- [6- (4-methylpiperidin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl } -1H-pyrrol-3-yl) acetic acid 54 mg was obtained as a cream solid (yield 59%).
MS (ESI): 416 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 12.31 (1H, brs), 9.03 (1H, s), 8.88 (1H, s), 8.33 (1H, s), 8.08 (1H, brs), 7.76 (1H, dd, J = 2.6,2.6), 7.71-7.69 (2H, m), 7.16 (1H, dd, J = 9.0,2.0), 6.40-6.39 (1H, m), 3.88 (2H, d, J = 12.0), 3.47 (2H, s), 2.97-2.93 (2H, m), 1.77-1.74 (2H, m), 1.61 (1H, m), 1.31 (2H, dq, J = 12.0, 3.4), 0.96 (3H, d, J = 6.6).

Reference Example 14
Ethyl 2- (1- {6- [6- (4-methylpiperazin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate

Under argon atmosphere, ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate 22 mg, copper iodide 1.0 mg, tripotassium phosphate n-hydrate 34 mg, trans-N, N'-dimethyl-1,2-cyclohexanediamine 1.7 mg and 6- (4-methylpiperazin-1-yl) -1H-indazole 16 mg are suspended in 0.62 mL of 1,4-dioxane and stirred at 110 ° C. for 24 hours. did. The reaction mixture was filtered through a celite bed, and the solvent was distilled off under reduced pressure. The obtained residue was purified by column chromatography (NH silica, ethyl acetate / cyclohexane = 2/1) to give ethyl 2- (1- {6- [6- (4-methylpiperazin-1-yl)- 15 mg of 1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate was obtained as a white solid (57% yield).
MS (ESI): 446 (M + H) ^+
1 H-NMR (400 MHz, CDCl ₃ ) δ: 9.16 (1H, s), 8.48 (1H, s), 8.20 (1H, s), 8.10 (1H, s), 7.63 (1H, d, J = 8.8) , 7.56-7.55 (2H, m), 7.07 (1H, dd, J = 9.1,2,1), 6.42 (1H, m), 4.20 (2H, q, J = 7.1), 3.57 (2H, s), 3.43-3.41 (4H, m), 2.66-2.64 (4H, m), 2.39 (3H, s), 1.30 (3H, t, J = 7.1).

Reference Example 15
2- (1- {6- [6- (4-Methylpiperazin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetic acid

Ethyl 2- (1- {6- [6- (4-methylpiperazin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H dissolved in 2.0 mL of 1,4-dioxane -Pyrrol-3-yl) acetate (17 mg) was added with 1N aqueous sodium hydroxide (0.40 mL) and stirred at room temperature for 21 hours. 1N Hydrochloric acid was added to the reaction mixture, and the mixture was concentrated under reduced pressure. The obtained residue was purified by column chromatography (reverse phase, water to methanol) to give 2- (1- {6- [6- (4-methylpiperazin-1-yl) -1H-indazol-1-yl ] 11 mg of pyrazin-2-yl} -1H-pyrrol-3-yl) acetic acid was obtained as a cream colored solid (68% yield).
MS (ESI): 418 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.01 (1H, s), 8.87 (1H, s), 8.35 (1H, s), 8.10 (1H, s), 7.74-7.68 (3H, m) , 7.19 (1H, dd, J = 8.8,2.1), 6.38 (1H, m), 3.37-3.33 (4H, m), 3.16 (2H, s), 2.67-2.50 (5H, m), 1.88 (3H, s).

Reference Example 16
Ethyl 2- (1- {6- [6- (4-methoxypiperidin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate

Under an argon atmosphere, ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate 100 mg, copper iodide 2.7 mg, tripotassium phosphate n-hydrate 156 mg, trans-N, N'-dimethyl-1,2-cyclohexanediamine 7.9 mg and 6- (4-methoxypiperidin-1-yl) -1H-indazole 78 mg were suspended in 1.11 mL of 1,4-dioxane and stirred at 110 ° C. for 24 hours. did. The reaction mixture was filtered through a celite bed, and the solvent was distilled off under reduced pressure. The obtained residue was purified by column chromatography (silica gel, ethyl acetate / cyclohexane = 1/5) to give ethyl 2- (1- {6- [6- (4-methoxypiperidin-1-yl) -1H -Indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate was obtained as a white solid, 50 mg (39% yield).
MS (ESI): 461 (M + H) ^+
1 H-NMR (400 MHz, CDCl ₃ ) δ: 9.11 (1H, s), 8.43 (1H, s), 8.14 (1H, s), 8.04 (1H, s), 7.57-7.51 (3H, m), 7.02 (1H, dd, J = 8.8,2.0), 6.42-6.41 (1H, m), 4.20 (2H, q, J = 7.1), 3.72-3.67 (2H, m), 3.56 (2H, s), 3.46- 3.43 (1H, m), 3.41 (3H, s), 3.17-3.11 (2H, m), 2.09-2.04 (2H, m), 1.81-1.74 (2H, m), 1.30 (3H, t, J = 7.1 ) ..

Reference Example 17
2- (1- {6- [6- (4-Methoxypiperidin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetic acid

Ethyl 2- (1- {6- [6- (4-methoxypiperidin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H dissolved in 2.0 mL of 1,4-dioxane -Pyrrol-3-yl) acetate (50 mg) was added with 1N aqueous sodium hydroxide solution (0.5 mL) and stirred at room temperature for 17 hours. 1N hydrochloric acid was added to the reaction solution. The precipitated precipitate was collected by filtration and washed with water to give 2- (1- {6- [6- (4-methoxypiperidin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl } -1H-pyrrol-3-yl) acetic acid (37 mg) was obtained as a white solid (yield 78%).
MS (ESI): 433 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.01 (1H, s), 8.87 (1H, s), 8.33 (1H, s), 8.09 (1H, brs), 7.75-7.69 (3H, m) , 7.17 (1H, dd, J = 8.8,1.8), 6.39-6.38 (1H, m), 3.69-3.66 (2H, m), 3.45-3.42 (3H, m), 3.28 (3H, s), 3.15- 3.10 (2H, m), 2.02-1.99 (2H, m), 1.62-1.57 (2H, m).

Reference Example 18
Ethyl 2- [1- (6- {6- [4- (hydroxymethyl) piperidin-1-yl] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetate

Under argon atmosphere, ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate 104 mg, tris (dibenzylideneacetone) dipalladium 13 mg, Xantphos 25 mg, cesium carbonate 227 mg, [1- 74 mg of (1H-indazol-6-yl) piperidin-4-yl] methanol was suspended in 2.32 mL of 1,4-dioxane and stirred at 110 ° C. for 24 hours. The reaction mixture was filtered through celite bed (THF), and the solvent was distilled off under reduced pressure. The obtained residue was purified by column chromatography (silica gel, ethyl acetate / cyclohexane = 1 / 2-2 / 1) to give ethyl 2- [1- (6- {6- [4- (hydroxymethyl) piperidine]. 1-yl] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetate was obtained as a pale yellow solid (81 mg, 61% yield).
MS (ESI): 461 (M + H) ^+
1 H-NMR (400 MHz, CDCl ₃ ) δ: 9.16 (1H, s), 8.48 (1H, s), 8.22 (1H, s), 8.10 (1H, s), 7.62 (1H, d, J = 8.8) , 7.57-7.54 (2H, m), 7.07 (1H, dd, J = 8.7,2.2), 6.44-6.43 (1H, m), 4.20 (2H, q, J = 7.1Hz), 3.96-3.93 (2H, m), 3.60-3.57 (4H, m), 2.97 (2H, ddd, J = 12.2,2.6,2.6), 1.94-1.91 (2H, m), 1.80 (1H, m), 1.52-1.45 (2H, m ), 1.29 (3H, t, J = 7.1).

Reference Example 19
2- [1- (6- {6- [4- (Hydroxymethyl) piperidin-1-yl] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetic acid

Ethyl 2- [1- (6- {6- [4- (hydroxymethyl) piperidin-1-yl] -1H-indazol-1-yl} pyrazin-2-yl dissolved in 3.0 mL of 1,4-dioxane ) -1H-pyrrol-3-yl] acetate (40 mg) was added 1N aqueous sodium hydroxide solution (1.0 mL), and the mixture was stirred at room temperature for 2 hr. 1N hydrochloric acid was added to the reaction solution. The precipitated precipitate was collected by filtration and washed with water to give 2- [1- (6- {6- [4- (hydroxymethyl) piperidin-1-yl] -1H-indazol-1-yl} pyrazine- 23 mg of 2-yl) -1H-pyrrol-3-yl] acetic acid was obtained as a pale yellow solid (62% yield).
MS (ESI): 433 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.02 (1H, s), 8.87 (1H, s), 8.32 (1H, s), 8.07 (1H, brs), 7.76-7.75 (1H, m) , 7.07 (1H, s), 7.69 (1H, d, J = 8.8), 7.16 (1H, dd, J = 9.0,2.0), 6.39-6.38 (1H, m), 3.93-3.90 (2H, m), 3.55 (3H, s), 3.45 (2H, s), 2.92-2.88 (2H, m), 1.80-1.77 (2H, m), 1.62 (1H, m), 1.36-1.27 (2H, m).

Reference Example 20
Ethyl 2- (1- {6- [6- (ethylamino) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate

Ethyl 2- {1- [6- (6-amino-1H-indazolyl) pyrazin-2-yl] pyrrol-3-yl} acetate (Reference Example 58 compound) 50 mg, sodium cyanoborohydride 43 mg, acetaldehyde 0.039 mL The residue was dissolved in 0.2 mL of acetic acid and 2 mL of N, N′-dimethylformamide and stirred at 50 ° C. for 18 hours. Saturated aqueous sodium hydrogen carbonate was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (n-hexane / ethyl acetate = 4/6) to give 27 mg of ethyl 2- (1- {6- [6- (Ethylamino) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate was obtained (yield 50%).
MS (ESI): 391 (M + H) ^<+> .

Reference Example 21
2- (1- {6- [6- (Ethylamino) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetic acid

Ethyl 2- (1- {6- [6- (ethylamino) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate 27 mg to 1,4-dioxane 2 mL After dissolution, 0.5 mL of 1M aqueous sodium hydroxide solution was added, and the mixture was stirred at room temperature for 2 hours. The reaction solution was adjusted to neutrality by adding 1M hydrochloric acid, and extracted with ethyl acetate. The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and 10 mg of 2- (1- {6- [6- (ethylamino) -1H-indazol-1-yl] pyrazine -2-yl} -1H-pyrrol-3-yl) acetic acid was obtained (38% yield).
MS (ESI): 363 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 1.29 (3H, t, J = 7.0), 3.21-3.24 (2H, m), 3.48 (2H, s), 6.40 (1H, brs), 6.43 ( 1H, brs), 6.76 (1H, d, J = 8.8), 7.57 (1H, d, J = 8.8), 7.65 (1H, s), 7.73 (1H, s), 7.78 (1H, s), 8.24 ( 1H, s), 8.86 (1H, s), 9.02 (1H, s).

Reference Example 22
Ethyl 2- (1- {6- [6- (hexylamino) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate

Ethyl 2- {1- [6- (6-amino-1H-indazolyl) pyrazin-2-yl] pyrrol-3-yl} acetate (Reference Example 58 compound) 50 mg, sodium cyanoborohydride 43 mg, 1-hexanal 69 mg Was dissolved in 0.2 mL of acetic acid and 2 mL of N, N′-dimethylformamide and stirred at 50 ° C. for 18 hours. Saturated aqueous sodium hydrogen carbonate was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (n-hexane / ethyl acetate = 7/3) to give 58 mg of ethyl 2- (1- {6- [6- (hexylamino) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate was obtained (95% yield).
MS (ESI): 447 (M + H) ^<+> .

Reference Example 23
2- (1- {6- [6- (hexylamino) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetic acid

58 mg of ethyl 2- (1- {6- [6- (hexylamino) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate in 2 mL of 1,4-dioxane After dissolution, 0.5 mL of 1M aqueous sodium hydroxide solution was added, and the mixture was stirred at room temperature for 2 hours. The reaction solution was adjusted to neutrality by adding 1M hydrochloric acid, and extracted with ethyl acetate. The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated and 48 mg of 2- (1- {6- [6- (hexylamino) -1H-indazol-1-yl] pyrazine -2-yl} -1H-pyrrol-3-yl) acetic acid was obtained (yield 88%).
MS (ESI): 419 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 0.87 (3H, t, J = 7.1), 1.28-1.36 (4H, m), 1.40-1.46 (2H, m), 1.65-1.72 (2H, m ), 3.15-3.22 (2H, m), 3.48 (2H, s), 6.38 (1H, brs), 6.43 (1H, brs), 6.78 (1H, dd, J = 1.9, 8.8), 7.56 (1H, d , J = 8.5), 7.63 (1H, s), 7.73 (1H, s), 7.78 (1H, t, J = 2.4), 8.23 (1H, s), 8.86 (1H, s), 9.02 (1H, s ) ..

Reference Example 24
Ethyl 2- (1- {6- [6- (2,6-dimethylmorpholin-4-yl) indazoyl] pyrazin-2-yl} pyrrol-3-yl) acetate

Under an argon atmosphere, trans-N, N′-dimethyl-1,2-cyclohexanediamine 8 mg, copper iodide 5 mg, ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate 65 mg 178 mg of tripotassium phosphate n-hydrate and 100 mg of 6- (2,6-dimethylmorpholin-4-yl) indazole were suspended in 1.4 mL of toluene and stirred at 110 ° C. for 24 hours. The reaction mixture was filtered through silica gel (ethyl acetate) and concentrated. The residue was purified by silica gel column chromatography (n-hexane / ethyl acetate = 1/1), and 30 mg of ethyl 2- (1- {6- [6- (2,6-dimethylmorpholin-4-yl) indazoyl] Pyrazin-2-yl} pyrrol-3-yl) acetate was obtained (23% yield).
MS (ESI): 460 (M + H) ^+
1 H-NMR (400 MHz, CDCl ₃ ) δ: 1.20-1.40 (9H, m), 2.45-2.64 (2H, m), 3.51 (1H, d, J = 11.2), 3.64 (1H, s), 3.66 ( 1H, d, J = 10.8), 3.80-3.95 (2H, m), 4.17-4.23 (2H, m), 6.41 (1H, s), 6.81 (1H, s), 6.85-6.94 (1H, m), 7.55-7.68 (3H.m), 7.93 (1H, s), 8.14 (1H, s), 8.49 (1H, s), 8.18 (1H, s).

Reference Example 25
2- (1- {6- [6- (2,6-Dimethylmorpholin-4-yl) indazoyl] pyrazin-2-yl} pyrrol-3-yl) acetic acid

Dissolve 30 mg of ethyl 2- (1- {6- [6- (2,6-dimethylmorpholin-4-yl) indazoyl] pyrazin-2-yl} pyrrol-3-yl) acetate in 1.0 mL of 1,4-dioxane Then, 0.3 mL of 1M aqueous sodium hydroxide solution was added, and the mixture was stirred overnight at room temperature. To the reaction solution was added 1M hydrochloric acid, and the mixture was extracted with ethyl acetate. The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and 19 mg of 2- (1- {6- [6- (2,6-dimethylmorpholin-4-yl) indazoyl] Pyrazin-2-yl} pyrrol-3-yl) acetic acid was obtained (68% yield).
MS (ESI): 432 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 1.19 (6H, d, J = 6.0), 2.30-2.50 (2H, s), 3.20-3.42 (2H, s), 3.46 (2H, s), 3.70-3.82 (2H, s), 6.36 (1H, s), 7.21 (1H, d, J = 8.8), 7.74-7.76 (3H, m), 8.03 (1H, s), 8.34 (1H, s), 8.86 (1H, s), 9.03 (1H, s).

Reference Example 26
Ethyl 2- (1- {6- [6- (3,5-dimethylpiperidin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate

Under an argon atmosphere, ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate 51 mg, copper iodide 1.4 mg, tripotassium phosphate n-hydrate 79 mg, trans-N, N′-dimethyl-1,2-cyclohexanediamine 4.0 mg, 6- (3,5-dimethylpiperidin-1-yl) -1H-indazole 39 mg was suspended in 1,4-dioxane 1.13 mL, and the mixture was stirred at 110 ° C. for 24 hours. Stir for hours. The reaction mixture was filtered through celite bed (THF), and the solvent was distilled off under reduced pressure. The obtained residue was purified by column chromatography (silica gel, ethyl acetate / cyclohexane = 1/4) to give ethyl 2- (1- {6- [6- (3,5-dimethylpiperidin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate was obtained as a yellow solid, 52 mg (yield 80%).
MS (ESI): 458 (M + H) ⁺ .
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 9.16 (1H, s), 8.46 (1H, s), 8.16 (1H, d, J = 2.0), 8.07 (1H, s), 7.64 (1H, dd, J = 2.9,2.9), 7.58 (1H, d, J = 9.0), 7.54-7.53 (1H, m), 7.47 (1H, m), 7.07-7.03 (1H, m), 6.40-6.40 (1H, m ), 4.20 (2H, q, J = 7.1), 3.82-3.80 (2H, m), 3.58 (2H, s), 2.38 (2H, t, J = 12.0), 1.88-1.86 (3H, m), 1.29 (3H, t, J = 7.1), 1.00 (6H, d, J = 4.7).

Reference Example 27
2- (1- {6- [6- (3,5-Dimethylpiperidin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetic acid

Ethyl 2- (1- {6- [6- (3,5-dimethylpiperidin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} dissolved in 4.0 mL of 1,4-dioxane} -1H-pyrrol-3-yl) acetate (50 mg) was added with 1N aqueous sodium hydroxide solution (0.21 mL), and the mixture was stirred at room temperature for 48 hours. 1N hydrochloric acid was added to the reaction solution. The precipitated precipitate was collected by filtration and washed with water to give 2- (1- {6- [6- (3,5-dimethylpiperidin-1-yl) -1H-indazol-1-yl] pyrazin-2 -Il} -1H-pyrrol-3-yl) acetic acid was obtained as a cream solid, 28 mg (yield 60%).
MS (ESI): 430 (M + H) ⁺ .
¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 12.34 (1H, brs), 9.03 (1H, s), 8.87 (1H, s), 8.32 (1H, s), 8.04 (1H, brs), 7.76 -7.68 (3H, s), 7.18 (1H, d, J = 8.8), 6.36 (1H, brs), 3.86-3.82 (2H, m), 3.48 (2H, s), 2.36 (2H, t, J = 12.0), 1.82-1.78 (3H, m), 0.94 (6H, d, J = 6.4), 0.77 (1H, q, J = 12.0).

Reference Example 28
Benzyl 4- (1- {6- [3- (2-ethoxy-2-oxoethyl) -1H-pyrrol-1-yl] pyrazin-2-yl} -1H-indazol-6-yl) piperazine-1-carboxy rate

Under argon atmosphere, ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate 300 mg, tris (dibenzylideneacetone) dipalladium 15 mg, Xantphos 29 mg, cesium carbonate 384 mg, benzyl 4- (1H -Indazol-6-yl) piperazine-1-carboxylate (340 mg) was suspended in 1,4-dioxane (6.80 mL) and stirred at 110 ° C. for 24 hours. The reaction mixture was filtered through a celite bed, and the solvent was distilled off under reduced pressure. Ethyl acetate was added to the resulting residue and purified by a reprecipitation method to obtain benzyl 4- (1- {6- [3- (2-ethoxy-2-oxoethyl) -1H-pyrrol-1-yl] pyrazine. 356 mg of 2-yl} -1H-indazol-6-yl) piperazine-1-carboxylate as a beige solid was obtained (yield 75%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 9.17 (1H, s), 8.50 (1H, s), 8.22 (1H, s), 8.12 (1H, s), 7.66 (1H, d, J = 9.1) 7.57 (1H, brs), 7.53-7.52 (1H, m), 7.39-7.33 (5H, m), 7.05 (1H, dd, J = 8.8,2.1), 6.42-6.41 (1H, m), 5.19 ( 2H, s), 4.17 (2H, q, J = 7.1), 3.77-3.74 (4H, m), 3.55 (2H, s), 3.38 (4H, brs), 1.27 (3H, t, J = 7.1). .

Reference Example 29
Ethyl 2- (1- {6- [6- (piperazin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate

Benzyl 4- (1- {6- [3- (2-ethoxy-2-oxoethyl) -1H-pyrrol-1-yl] pyrazin-2-yl} -1H dissolved in a mixed solvent of 25 mL of ethanol and 10 mL of tetrahydrofuran -Indazol-6-yl) piperazine-1-carboxylate (100 mg) was mixed with ammonium formate (55 mg) and 10% palladium / carbon (20 mg), and heated under reflux for 30 minutes. The reaction mixture was filtered through celite bed (THF), and the solvent was distilled off under reduced pressure. The obtained residue was purified by column chromatography (silica gel, chloroform / methanol = 300/1 to 10/1) to give ethyl 2- (1- {6- [6- (piperazin-1-yl) -1H -Indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate was obtained in an amount of 70 mg (93% yield).
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 9.16 (1H, s), 8.48 (1H, s), 8.20 (1H, s), 8.10 (1H, s), 7.63 (1H, d, J = 7.0) , 7.55-7.54 (2H, m), 7.06 (1H, dd, J = 8.8,2.1), 6.42 (1H, m), 4.20 (2H, q, J = 7.1), 3.56 (2H, s), 3.37- 3.34 (4H, m), 3.13-3.10 (4H, m), 1.29 (3H, t, J = 7.1).

Reference Example 30
Ethyl 2- (1- {6- [6- (4-isopropylpiperazin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate

Ethyl 2- (1- {6- [6- (piperazin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate dissolved in 1.0 mL DMF To 50 mg, 0.043 mL of acetone, 0.1 mL of acetic acid and 36 mg of sodium cyanoborohydride were added and stirred at 60 ° C. for 9 hours. Saturated aqueous sodium hydrogen carbonate was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The solvent was distilled off under reduced pressure, and the resulting residue was purified by column chromatography (amine silica, ethyl acetate / cyclohexane = 1/3) to give ethyl 2- (1- {6- [6- (4-isopropylpiperazine). 36 mg of 1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate was obtained as a white solid (67% yield).
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 9.16 (1H, s), 8.48 (1H, s), 8.20 (1H, s), 8.10 (1H, s), 7.63 (1H, d, J = 8.8) 7.56-7.56 (2H, m), 7.07 (1H, dd, J = 8.8,2.1), 6.43-6.42 (1H, m), 4.20 (2H, q, J = 7.1), 3.57 (2H, s), 3.43-3.41 (4H, m), 2.77-2.75 (4H, m), 1.29 (3H, t, J = 7.1), 1.12 (6H, d, J = 4.6).

Reference Example 31
2- (1- {6- [6- (4-Isopropylpiperazin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetic acid

Ethyl 2- (1- {6- [6- (4-isopropylpiperazin-1-yl) -1H-indazol-1-yl] pyrazin-2-yl} -1H dissolved in 2.0 mL of 1,4-dioxane -Pyrrol-3-yl) acetate (35 mg) was added with 1 N aqueous sodium hydroxide (1.0 mL) and stirred at room temperature for 7 hours. 1N Hydrochloric acid was added to the reaction mixture, and the mixture was concentrated under reduced pressure. The obtained residue was purified by column chromatography (reverse phase, water to methanol) to give 2- (1- {6- [6- (4-isopropylpiperazin-1-yl) -1H-indazol-1-yl ] 32 mg of pyrazin-2-yl} -1H-pyrrol-3-yl) acetic acid was obtained as a pale yellow solid (quantitative).
¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.00 (1H, s), 8.86 (1H, s), 8.33 (1H, s), 8.08 (1H, s), 7.74-7.70 (3H, m) , 7.16 (1H, d, J = 9.0), 6.38 (1H, brs), 3.43 (2H, s), 3.32 (4H, m), 2.74-2.71 (1H, m), 2.67 (4H, m), 1.03 (6H, d, J = 6.5).

Reference Example 32
Ethyl 2- [1- (6- {6- [4- (2-methoxyethyl) piperazin-1-yl] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl ]acetate

Under argon atmosphere, ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate 91 mg, tris (dibenzylideneacetone) dipalladium 4.6 mg, Xantphos 8.9 mg, cesium carbonate 116 mg, 6 80 mg of [4- (2-methoxyethyl) piperazin-1-yl] -1H-indazole was suspended in 1.02 mL of 1,4-dioxane, and stirred at 110 ° C. for 24 hours. The reaction mixture was filtered through celite bed (THF), and the solvent was distilled off under reduced pressure. The obtained residue was purified by column chromatography (silica gel, ethyl acetate) to give ethyl 2- [1- (6- {6- [4- (2-methoxyethyl) piperazin-1-yl] -1H- 102 mg of indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetate was obtained as a pale yellow solid (81% yield).
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 9.16 (1H, s), 8.48 (1H, s), 8.20 (1H, s), 8.10 (1H, s), 7.63 (1H, d, J = 9.0) , 7.57-7.54 (2H, m), 7.06 (1H, dd, J = 8.8,2.0), 6.42 (1H, m), 4.20 (2H, q, J = 7.1), 3.59 (2H, t, J = 5.4 ), 3.57 (2H, s), 3.45-3.43 (4H, m), 3.39 (3H, s), 2.75-2.73 (4H, m), 2.68 (2H, t, J = 5.4), 1.30 (3H, t , J = 7.1).

Reference Example 33
2- [1- (6- {6- [4- (2-methoxyethyl) piperazin-1-yl] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] Acetic acid

Ethyl 2- [1- (6- {6- [4- (2-methoxyethyl) piperazin-1-yl] -1H-indazol-1-yl) dissolved in 4.0 mL 1,4-dioxane and 2.0 mL methanol } To 100 mg of pyrazin-2-yl) -1H-pyrrol-3-yl] acetate was added 1.0 mL of 1N aqueous sodium hydroxide solution, and the mixture was stirred at room temperature for 3 hours. 1N hydrochloric acid was added to the reaction solution. The precipitated precipitate was collected by filtration and washed with water to give 2- [1- (6- {6- [4- (2-methoxyethyl) piperazin-1-yl] -1H-indazol-1-yl} 85 mg of pyrazin-2-yl) -1H-pyrrol-3-yl] acetic acid was obtained (yield 90%).
¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.01 (1H, s), 8.87 (1H, s), 8.33 (1H, s), 8.06 (1H, brs), 7.75-7.72 (2H, m) , 7.70 (1H, s), 7.16 (1H, d, J = 8.8), 6.39-6.38 (1H, m), 3.49 (2H, t, J = 5.7), 3.45 (2H, s), 3.32 (4H, m), 3.25 (3H, s), 2.63 (4H, m), 2.55 (2H, t, J = 5.7).

Reference Example 34
Ethyl 2- [1- (6- {6- [4- (2-methoxyethoxy) piperidin-1-yl] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl ]acetate

Under argon atmosphere, ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate 47 mg, tris (dibenzylideneacetone) dipalladium 6.0 mg, Xantphos 11 mg, cesium carbonate 60 mg, 6- [ 40 mg of 4- (2-methoxyethoxy) piperidin-1-yl] -1H-indazole was suspended in 1.30 mL of 1,4-dioxane and stirred at 110 ° C. for 24 hours. The reaction mixture was filtered through celite bed (THF), and the solvent was distilled off under reduced pressure. The obtained residue was purified by column chromatography (silica gel, ethyl acetate / cyclohexane = 1/3 to 1/1) to give ethyl 2- [1- (6- {6- [4- (2-methoxyethoxy ) Piperidin-1-yl] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetate was obtained as a pale yellow solid (81 mg, yield 61%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 9.15 (1H, s), 8.47 (1H, s), 8.20 (1H, brs), 8.08 (1H, s), 7.60 (1H, d, J = 9.1) 7.55-7.54 (2H, m), 7.06 (1H, dd, J = 8.8,2.0), 6.43-6.42 (1H, m), 4.20 (2H, q, J = 7.1), 3.78-3.75 (2H, m ), 3.70-3.67 (2H, m), 3.60-3.57 (5H, m), 3.40 (3H, s), 3.16-3.10 (2H, m), 2.13-2.06 (2H, m), 1.86-1.78 (2H) , m), 1.30 (3H, t, J = 7.1).

Reference Example 35
2- [1- (6- {6- [4- (2-methoxyethoxy) piperidin-1-yl] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] Acetic acid

Ethyl 2- [1- (6- {6- [4- (2-methoxyethoxy) piperidin-1-yl] -1H-indazol-1-yl} pyrazin-2 dissolved in 4.0 mL of 1,4-dioxane -Yl) -1H-pyrrol-3-yl] acetate (57 mg) was added with 1N aqueous sodium hydroxide (4.0 mL) and stirred at room temperature for 20 hours. 1N Hydrochloric acid was added to the reaction mixture, and the aqueous layer was extracted with chloroform / methanol (4/1). The organic layer was dried over anhydrous sodium sulfate and the solvent was distilled off under reduced pressure to give 2- [1- (6- {6- [4- (2-methoxyethoxy) piperidin-1-yl] -1H-indazole-1 -Il} pyrazin-2-yl) -1H-pyrrol-3-yl] acetic acid 54 mg was obtained (quantitative).
MS (ESI): 476 (M + H) ⁺ .
¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.00 (1H, s), 8.85 (1H, s), 8.30 (1H, s), 8.06 (1H, s), 7.74 (1H, dd, J = 2.7, 2.4), 7.69-7.67 (2H, m), 7.15 (1H, dd, J = 8.8, 2.1), 6.39-6.38 (1H, m), 3.80-3.67 (3H, m), 3.58-3.52 (3H , m), 3.24 (3H, s), 3.11-3.06 (2H, m), 2.00-1.55 (6H, m).

Reference Example 36
Ethyl 2- [1- (6- {6- [4- (methoxymethyl) piperidin-1-yl] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetate

Under argon atmosphere, ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate 95 mg, tris (dibenzylideneacetone) dipalladium 12 mg, Xantphos 23 mg, cesium carbonate 122 mg, 6- [4- 72 mg of (methoxymethyl) piperidin-1-yl] -1H-indazole was suspended in 2.60 mL of 1,4-dioxane and stirred at 110 ° C. for 24 hours. The reaction mixture was filtered through celite bed (THF), and the solvent was distilled off under reduced pressure. The obtained residue was purified by column chromatography (silica gel, ethyl acetate / cyclohexane = 1/4) to give ethyl 2- [1- (6- {6- [4- (methoxymethyl) piperidin-1-yl]. ] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetate was obtained as an ocherous solid (113 mg, yield 90%).
¹ H-NMR (400 MHz, CDCl ₃ ) δ: 9.16 (1H, s), 8.47 (1H, s), 8.20 (1H, s), 8.09 (1H, s), 7.61 (1H, d, J = 9.0) 7.57-7.55 (2H, m), 7.07 (1H, dd, J = 8.8,2.1), 6.43-6.42 (1H, m), 4.20 (2H, q, J = 7.1), 3.94-3.91 (2H, m ), 3.57 (2H, s), 3.37 (3H, s), 3.30 (2H, d, J = 6.4), 2.96-2.90 (2H, m), 1.92-181 (3H, m), 1.56 (3H, s) ), 1.53-1.42 (1H, m), 1.30 (3H, t, J = 7.1).

Reference Example 37
2- [1- (6- {6- [4- (Methoxymethyl) piperidin-1-yl] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetic acid

Ethyl 2- [1- (6- {6- [4- (methoxymethyl) piperidin-1-yl] -1H-indazol-1-yl} pyrazin-2-yl) dissolved in 5.0 mL of 1,4-dioxane ) -1H-pyrrol-3-yl] acetate (110 mg) was added 1N aqueous sodium hydroxide solution (5.0 mL), and the mixture was stirred at room temperature for 26 hr. 1N hydrochloric acid was added to the reaction solution. The precipitated precipitate was collected by filtration and washed with water to give 2- [1- (6- {6- [4- (methoxymethyl) piperidin-1-yl] -1H-indazol-1-yl} pyrazine- 105 mg of 2-yl) -1H-pyrrol-3-yl] acetic acid were obtained (quantitative).
¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 9.02 (1H, s), 8.87 (1H, s), 8.32 (1H, s), 8.07 (1H, brs), 7.75 (1H, dd, J = 2.7, 2.7), 7.70-7.68 (2H, m), 7.15 (1H, dd, J = 8.8, 2.2), 6.39-6.37 (1H, m), 3.92-3.88 (2H, m), 3.45 (2H, s ), 3.23 (3H, s), 3.23-3.21 (2H, m), 2.94-2.87 (2H, m), 1.79-1.76 (3H, m), 1.39-1.31 (2H, m).

Reference Example 38
Ethyl 2- [1- (6- {6- [N- (2-morpholinoethyl) methane-6-ylsulfonamido] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrole-3 -Il] acetate

Under argon atmosphere, trans-N, N'-dimethyl-1,2-cyclohexanediamine 12 mg, copper iodide 4 mg, N- (1H-indazol-6-yl) -N- (2-morpholinoethyl) methanesulfonamide 150 mg , 187 mg of tripotassium phosphate n-hydrate and 150 mg of ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate were suspended in 1 mL of toluene and stirred at 110 ° C. for 24 hours. did. The reaction mixture was filtered through silica gel (ethyl acetate) and concentrated. The residue was purified by silica gel column chromatography (NH silica gel, n-hexane / ethyl acetate = 1/1), and 202 mg of ethyl 2- [1- (6- {6- [N- (2-morpholinoethyl) methane- 6-ylsulfonamido] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetate was obtained (yield 87%).
MS (ESI): 554 (M + H) ^+
1 H-NMR (400 MHz, CDCl ₃ ) δ: 1.31 (3H, t, J = 7.3), 2.40-2.44 (4H, m), 2.53 (2H, t, J = 6.6), 3.07 (3H, s), 3.56-3.62 (6H, m), 3.97 (2H, t, J = 6.6), 4.21 (2H, q, J = 7.3), 6.44 (1H, brs), 7.38 (1H, d, J = 8.5), 7.54 (1H, brs), 7.61 (1H, s), 7.84 (1H, d, J = 8.5), 8.29 (1H, s), 8.56 (1H, s), 8.82 (1H, s), 9.18 (1H, s ) ..

Reference Example 39
2- [1- (6- {6- [N- (2-morpholinoethyl) methane-6-ylsulfonamido] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrole-3- Il] acetic acid

Ethyl 2- [1- (6- {6- [N- (2-morpholinoethyl) methane-6-ylsulfonamido] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrole-3 -Ill] acetate (202 mg) was dissolved in 1,4-dioxane (3 mL), 1 M aqueous sodium hydroxide solution (1.5 mL) was added, and the mixture was stirred at room temperature for 2 hr. The reaction solution was adjusted to neutrality by adding 1M hydrochloric acid, and extracted with ethyl acetate. The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and 118 mg of 2- [1- (6- {6- [N- (2-morpholinoethyl) methane-6-yl]. Sulfonamido] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetic acid was obtained (yield 62%).
MS (ESI): 526 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 2.30 (4H, brs), 2.41 (2H, t, J = 6.6), 3.13 (4H, brs), 3.48 (2H, s), 3.91 (2H, t, J = 6.6), 6.40 (1H, brs), 7.49 (1H, d, J = 8.9), 7.75 (1H, brs), 7.79 (1H, s), 8.02 (1H, d, J = 8.5), 8.62 (1H, s), 8.68 (1H, s), 8.97 (1H, s), 9.07 (1H, s).

Reference Example 40
Ethyl 2- (1- {6- [6- (methylsulfonamido) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate

Ethyl 2- {1- [6- (6-amino-1H-indazolyl) pyrazin-2-yl] pyrrol-3-yl} acetate (Compound of Reference Example 58) 50 mg is dissolved in 3 mL of methylene chloride, and methanesulfonyl chloride 17 mg 4- (dimethylamino) pyridine (21 mg) was added, and the mixture was stirred at room temperature for 2 hours. 1M Hydrochloric acid was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (n-hexane / ethyl acetate = 3/1), and 47 mg of ethyl 2- (1- {6- [6- (methylsulfonamido) -1H-indazol-1-yl] Pyrazin-2-yl} -1H-pyrrol-3-yl) acetate was obtained (78% yield).
MS (ESI): 441 (M + H) ^+
1 H-NMR (400 MHz, CDCl ₃ ) δ: 1.33 (3H, t, J = 7.3), 3.11 (3H, s), 3.67 (2H, s), 4.30 (2H, q, J = 7.3), 6.40 ( 1H, brs), 7.30-7.33 (2H, m), 7.46 (1H, t, J = 2.7), 7.78 (1H, d, J = 8.5), 7.99 (1H, s), 8.23 (1H, s), 8.53 (1H, s), 8.74 (1H, s), 9.16 (1H, s).

Reference Example 41
2- (1- {6- [6- (Methylsulfonamido) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetic acid

Ethyl 2- (1- {6- [6- (methylsulfonamido) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate 48 mg, 1,4-dioxane 2 mL 1M sodium hydroxide aqueous solution 0.5mL was added, and it stirred at room temperature for 2 hours. 1M Hydrochloric acid was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and 11 mg of 2- (1- {6- [6- (methylsulfonamido) -1H-indazol-1-yl] Pyrazin-2-yl} -1H-pyrrol-3-yl) acetic acid was obtained (yield 25%).
MS (ESI): 413 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 3.11 (3H, s), 3.49 (2H, s), 6.39 (1H, brs), 7.18 (1H, d, J = 8.5), 7.85-7.91 ( 3H, m), 8.50 (1H, s), 8.66 (1H, s), 8.92 (1H, s), 9.07 (1H, s).

Reference Example 42
Ethyl 2- (1- {6- [6- (N-ethylmethane-6-ylsulfonamido) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate

Under argon atmosphere, trans-N, N'-dimethyl-1,2-cyclohexanediamine 13 mg, copper iodide 4 mg, N-ethyl-N- (1H-indazol-6-yl) methanesulfonamide 118 mg, tripotassium phosphate -200 mg of n hydrate and 160 mg of ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate were suspended in 1 mL of toluene and stirred at 110 ° C. for 24 hours. The reaction mixture was filtered through silica gel (ethyl acetate) and concentrated. The residue was purified by silica gel column chromatography (n-hexane / ethyl acetate = 1/1), and 152 mg of ethyl 2- (1- {6- [6- (N-ethylmethane-6-ylsulfonamide) -1H- Indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate was obtained (72% yield).
MS (ESI): 469 (M + H) ^+
1 H-NMR (400 MHz, CDCl ₃ ) δ: 1.23 (3H, t, J = 7.1) 1.32 (3H, t, J = 7.1), 2.99 (3H, s), 3.61 (2H, s), 3.07 (3H , s), 3.92 (2H, q, J = 7.1), 4.22 (2H, q, J = 7.3), 6.43 (1H, brs), 7.37 (1H, dd, J = 1.7, 8.5), 7.55 (1H, t, J = 2.69), 7.63 (1H, brs), 7.85 (1H, d, J = 8.5), 8.30 (1H, s), 8.57 (1H, s), 8.81 (1H, s), 9.19 (1H, s).

Reference Example 43
2- (1- {6- [6- (N-Ethylmethane-6-ylsulfonamido) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetic acid

152 mg of ethyl 2- (1- {6- [6- (N-ethylmethane-6-ylsulfonamido) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetate After dissolving in 4 mL of 1,4-dioxane, 1.5 mL of 1M aqueous sodium hydroxide solution was added, and the mixture was stirred at room temperature for 2 hours. 1M Hydrochloric acid was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and 129 mg of 2- (1- {6- [6- (N-ethylmethane-6-ylsulfonamido) -1H- Indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetic acid was obtained (90% yield).
MS (ESI): 441 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 1.10 (3H, t, J = 7.1) 3.09 (3H, s), 3.44 (sH, s), 3.85 (2H, q, J = 7.1), 6.40 (1H, brs), 7.45 (1H, dd, J = 1.7,8.5), 7.73 (1H, brs), 7.77 (1H, t, J = 2.9), 8.02 (1H, d, J = 8.5), 8.62 ( 1H, s), 8.65 (1H, s), 8.96 (1H, s), 9.06 (1H, s).

Reference Example 44
Ethyl 2- [1- (6- {6- [cyclopentyl (methyl) amino] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetate

Under argon atmosphere, trans-N, N'-dimethyl-1,2-cyclohexanediamine 12 mg, copper iodide 4 mg, N-cyclopentyl-N-methyl-1H-indazol-6-amine 118 mg, tripotassium phosphate / water 187 mg of a Japanese product and 150 mg of ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate were suspended in 1 mL of toluene and stirred at 110 ° C. for 24 hours. The reaction mixture was filtered through silica gel (ethyl acetate) and concentrated. The residue was purified by silica gel column chromatography (n-hexane / ethyl acetate = 3/1), and 155 mg of ethyl 2- [1- (6- {6- [cyclopentyl (methyl) amino] -1H-indazole-1- IL} pyrazin-2-yl) -1H-pyrrol-3-yl] acetate was obtained (yield 82%).
MS (ESI): 445 (M + H) ^+
1 H-NMR (400 MHz, CDCl ₃ ) δ: 1.31 (3H, t, J = 7.1), 1.54-1.60 (2H, m), 1.63-1.73 (2H, m), 1.76-1.84 (2H, m), 1.96-2.04 (2H, m), 2.98 (3H, s), 3.58 (2H, s), 4.21 (2H, q, J = 7.1), 4.38-4.43 (1H, m), 6.42 (1H, brs), 6.96 (1H, dd, J = 2.2,9.0), 7.53 (1H, s), 7.57-7.61 (2H, m), 8.05 (1H, s), 8.07 (1H, s), 8.47 (1H, s), 9.17 (1H, s).

Reference Example 45
2- [1- (6- {6- [Cyclopentyl (methyl) amino] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetic acid

Ethyl 2- (1- (6- {6- (cyclopentyl (methyl) amino) -1H-indazol-1-yl) pyrazin-2-yl) -1H-pyrrol-3-yl) acetate (154 mg) Dissolved in 2 mL of dioxane, 1.5 mL of 1M aqueous sodium hydroxide solution was added, and the mixture was stirred at room temperature for 2 hours. 1M Hydrochloric acid was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and 29 mg of 2- [1- (6- {6- [cyclopentyl (methyl) amino] -1H-indazole-1- Yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetic acid was obtained (yield 20%).
MS (ESI): 417 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 1.56-1.76 (6H, m), 1.85-1.95 (2H, m) 2.91 (3H, s), 3.46 (2H, s), 4.38-4.43 (1H , m), 6.37 (1H, brs), 7.03 (1H, d, J = 9.0), 7.66 (1H, d, J = 8.8), 7.70 (1H, m), 7.74 (1H, s), 7.91 (1H , s), 8.27 (1H, s), 8.84 (1H, s), 9.01 (1H, s).

Reference Example 46
Ethyl 2- [1- (6- {6- [3- (diethylamino) pyrrolidin-1-yl] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetate

Under an argon atmosphere, ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate 75 mg, tris (dibenzylideneacetone) dipalladium 5.8 mg, Xantphos 7.3 mg, cesium carbonate 96 mg, N, 60 mg of N-diethyl-1- (1H-indazol-6-yl) pyrrolidin-3-amine was suspended in 1.0 mL of 1,4-dioxane and stirred at 110 ° C. for 24 hours. The reaction mixture was filtered through celite, and the solvent was distilled off under reduced pressure. The residue obtained was purified by column chromatography (NH silica gel, n-hexane / ethyl acetate = 3/1) to give ethyl 2- [1- (6- {6- [3- (diethylamino) pyrrolidine-1 102 mg of (-yl) -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetate were obtained (yield 100%).
MS (ESI): 488 (M + H) ^+
1 H-NMR (400 MHz, CDCl ₃ ) δ: 1.09 (6H, t, J = 7.3), 1.30 (3H, t, J = 7.1), 1.98-2.05 (1H, m), 2.26-2.33 (1H, m ), 2.74 (1H, q, J = 7.3), 3.32 (1H, t, J = 8.5), 3.43-3.51 (2H, m), 3.57 (2H, s), 3.59-3.68 (2H, m), 4.20 (2H, q, J = 7.1), 6.39 (1H, brs), 6.69 (1H, dd, J = 1.7,9.0), 7.51 (1H, s), 7.59 (1H, d, J = 8.8), 7.62 ( 1H, t, J = 2.4), 7.78 (1H, s), 8.07 (1H, s), 8.45 (1H, s), 9.15 (1H, s).

Reference Example 47
2- [1- (6- {6- [3- (Diethylamino) pyrrolidin-1-yl] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetic acid

Ethyl 2- [1- (6- {6- [3- (diethylamino) pyrrolidin-1-yl] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetate 102 mg Was dissolved in 2 mL of 1,4-dioxane, 0.8 mL of 1M aqueous sodium hydroxide solution was added, and the mixture was stirred at room temperature for 2 hours. 1M Hydrochloric acid was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and 68 mg of 2- [1- (6- {6- [3- (diethylamino) pyrrolidin-1-yl] -1H -Indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetic acid was obtained (yield 71%).
MS (ESI): 460 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 1.09 (3H, t, J = 7.3), 2.02-2.08 (1Hm), 2.20-2.28 (1H, m), 2.65-2.85 (4H, m), 3.32-3.40 (4H, m), 3.57-3.65 (2H, m), 3.72-3.76 (1H, m), 6.32 (1H, brs), 6.77 (1H, d, J = 8.3), 7.65-7.70 (3H m), 7.75 (1H, s), 8.29 (1H, s), 8.84 (1H, s), 9.01 (1H, s).

Reference Example 48
Ethyl 2- [1- (6- {6- [3- (dimethylamino) pyrrolidin-1-yl] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetate

Under argon atmosphere, ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate 130mg, tris (dibenzylideneacetone) dipalladium 10mg, Xantphos 12.6mg, cesium carbonate 166mg, N, N 88-mg of dimethyl-1- (1H-indazol-6-yl) pyrrolidin-3-amine was suspended in 1.5 mL of 1,4-dioxane and stirred at 110 ° C. for 24 hours. The reaction mixture was filtered through celite, and the solvent was distilled off under reduced pressure. The residue obtained was purified by column chromatography (NH silica gel, n-hexane / ethyl acetate = 3/1) to give ethyl 2- [1- (6- {6- [3- (dimethylamino) pyrrolidine- 142 mg of 1-yl] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetate was obtained (yield 85%).
MS (ESI): 460 (M + H) ^+
1 H-NMR (400 MHz, CDCl ₃ ) δ: 1.30 (3H, t, J = 7.1), 1.67-1.74 (2H, m), 1.95-2.05 (1H, m), 2.28-2.32 (1H, m), 2.37 (6H, s), 2.88-2.95 (1H, m), 3.32 (1H, t, J = 8.3), 3.46-3.52 (1H, m), 3.56 (2H, s), 3.60-3.67 (1H, s ), 4.20 (2H, q, J = 7.1), 6.38 (1H, brs), 6.67 (1H, d, J = 8.8), 7.50 (1H, s), 7.57 (1H, d, J = 8.8), 7.61 (1H, t, J = 2.7), 7.76 (1H, s), 8.06 (1H, s), 8.44 (1H, s), 9.14 (1H, s).

Reference Example 49
2- [1- (6- {6- [3- (Dimethylamino) pyrrolidin-1-yl] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetic acid

Ethyl 2- [1- (6- {6- [3- (dimethylamino) pyrrolidin-1-yl] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetate 142 mg was dissolved in 2 mL of 1,4-dioxane, 0.8 mL of 1M aqueous sodium hydroxide solution was added, and the mixture was stirred at room temperature for 2 hours. 1M Hydrochloric acid was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and 116 mg of 2- [1- (6- {6- [3- (dimethylamino) pyrrolidin-1-yl]- 1H-Indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetic acid was obtained (88% yield).
MS (ESI): 432 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 2.01-2.10 (1H, m), 2.20-2.28 (1H, m), 2.36 (6H, s), 3.37-3.47 (7H, m), 6.32 ( 1H, brs), 6.78 (1H, dd, J = 2.2,8.8), 7.66-7.70 (3H, m), 7.77 (1H, d, J = 2.2), 8.30 (1H, s), 8.86 (1H, s ), 9.01 (1H, s).

Reference Example 50
Ethyl 2- [1- (6- {6- [4- (dimethylamino) piperidin-1-yl] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetate

Under an argon atmosphere, ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate 100 mg, tris (dibenzylideneacetone) dipalladium 7.7 mg, Xantphos 9.7 mg, cesium carbonate 128 mg, 1- 72 mg of (1H-indazol-6-yl) -N, N-dimethylpiperidin-4-amine was suspended in 1.5 mL of 1,4-dioxane and stirred at 110 ° C. for 24 hours. The reaction mixture was filtered through celite, and the solvent was distilled off under reduced pressure. The residue obtained was purified by column chromatography (NH silica gel, n-hexane / ethyl acetate = 1/1) to give ethyl 2- [1- (6- {6- [4- (dimethylamino) piperidine- 75 mg of 1-yl] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetate was obtained (57% yield).
MS (ESI): 474 (M + H) ^+
1 H-NMR (400 MHz, CDCl ₃ ) δ: 1.30 (3H, t, J = 7.1), 1.70-1.76 (2H, m), 1.97-2.05 (2H, m), 2.33-2.36 (7H, m), 2.94 (2H, t, J = 10.2), 3.57 (2H, s), 3.95 (2H, brs), 4.20 (2H, q, J = 7.1), 6.44 (1H, brs), 7.07 (1H, dd, J = 2.0,8.8), 7.55-7.58 (2H, m), 7.62 (1H, d, J = 9.0), 8.09 (1H, s), 8.21 (1H, s), 8.48 (1H, s), 9.16 (1H , s) ..

Reference Example 51
2- [1- (6- {6- [4- (Dimethylamino) piperidin-1-yl] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetic acid

Ethyl 2- [1- (6- {6- [4- (dimethylamino) piperidin-1-yl] -1H-indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetate 75 mg was dissolved in 2 mL of 1,4-dioxane, 0.5 mL of 1M aqueous sodium hydroxide solution was added, and the mixture was stirred at room temperature for 2 hours. 1M Hydrochloric acid was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and 59 mg of 2- [1- (6- {6- [4- (dimethylamino) piperidin-1-yl]- 1H-Indazol-1-yl} pyrazin-2-yl) -1H-pyrrol-3-yl] acetic acid was obtained (yield 84%).
MS (ESI): 446 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 1.58-1.64 (2H, m), 1.94-2.00 (2H, m), 2.35 (6H, s), 2.50-2.54 (1H, m), 2.92 ( 2H, t, J = 11.7), 3.47 (2H, s), 3.90-3.95 (2H, m), 6.40 (1H, brs), 7.17 (1H, dd, J = 2.0,8.8), 7.70-7.73 (2H , m), 7.76 (1H, t, J = 2.9), 8.09 (1H, s), 8.34 (1H, s), 8.88 (1H, s), 9.02 (1H, s).

Reference Example 52
Ethyl 2- (1- {6- [6- (5,6-dihydropyridin-1-yl) indazol-1-yl] pyrazin-2-yl} -pyrrol-3-yl) acetate

Under argon atmosphere, trans-N, N'-dimethyl-1,2-cyclohexanediamine 9 mg, copper iodide 6 mg, 6- (5,6-dihydropyridin-1-yl) indazole 66 mg, tripotassium phosphate / n hydrate 212 mg of the product and 119 mg of ethyl 2- [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate were suspended in 1.5 mL of toluene and stirred at 110 ° C. for 24 hours. The reaction mixture was filtered through silica gel (ethyl acetate) and concentrated. The residue was purified by silica gel column chromatography (n-hexane / ethyl acetate = 8/1), and 21 mg of ethyl 2- (1- {6- [6- (5,6-dihydropyridin-1-yl) indazole-1 -Il] pyrazin-2-yl} -pyrrol-3-yl) acetate was obtained (yield 15%).
MS (ESI): 429 (M + H) ^+
1 H-NMR (400 MHz, CDCl ₃ ) δ: 1.30 (3H, t, J = 7.1), 2.32-2.43 (2H, m), 3.52-3.61 (4H, m), 3.88-3.93 (2H, m), 4.20 (2H, q, J = 7.1), 5.50-6.00 (2H, m), 6.43-6.44 (1H, m), 7.07 (1H, dd, J = 2.2,9.0), 7.56-7.64 (3H, m) 8.10 (1H, s), 8.18 (1H, s), 8.48 (1H, s), 9.17 (1H, s).

Reference Example 53
2- (1- {6- [6- (5,6-Dihydropyridin-1-yl) indazol-1-yl] pyrazin-2-yl} -pyrrol-3-yl) acetic acid

Ethyl 2- (1- {6- [6- (5,6-dihydropyridin-1-yl) indazol-1-yl] pyrazin-2-yl} -pyrrol-3-yl) acetate 21 mg was added to 1,4-dioxane It melt | dissolved in 2 mL, 0.2 mL of 1M sodium hydroxide aqueous solution was added, and it stirred at room temperature for 2 hours. To the reaction solution was added 1M hydrochloric acid, and the mixture was extracted with ethyl acetate. The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and 4 mg of 2- (1- {6- [6- (5,6-dihydropyridin-1-yl) indazole-1 -Yl] pyrazin-2-yl} -pyrrol-3-yl) acetic acid was obtained (yield 23%).
MS (ESI): 401 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 2.23-2.33 (2H, m), 3.45-3.60 (4H, m), 3.80-3.88 (2H, m), 5.85-5.96 (2H, m), 6.37 (1H, s), 7.15-7.17 (1H, m), 7.67-7.72 (3H, m), 8.04 (1H, s), 8.31 (1H, s), 8.34 (1H, s), 8.99 (1H, s).

Reference Example 54
Ethyl 2- (1- {6- [6- (pyrrol-1-yl) -indazol-1-yl] pyrazin-2-yl} -pyrrol-3-yl) acetate

Under an argon atmosphere, trans-N, N'-dimethyl-1,2-cyclohexanediamine 5 mg, copper iodide 3 mg, 6-pyrrol-1-yl-indazole 30 mg, tripotassium phosphate n-hydrate 103 mg, ethyl 2 58 mg of [1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate was suspended in 1 mL of toluene and stirred at 110 ° C. for 24 hours. The reaction mixture was filtered through silica gel (ethyl acetate) and concentrated. The residue was purified by silica gel column chromatography (n-hexane / ethyl acetate = 10/1), and 56 mg of ethyl 2- (1- {6- [6- (pyrrol-1-yl) -indazol-1-yl] Pyrazin-2-yl} -pyrrol-3-yl) acetate was obtained (85% yield).
MS (ESI): 413 (M + H) ^+
1 H-NMR (400 MHz, CDCl ₃ ) δ: 1.30 (3H, t, J = 7.1), 3.60 (2H, s), 4.21 (2H, q, J = 7.1), 6.43-6.46 (3H, m), 7.23-7.32 (2H, m), 7.45-7.47 (1H, m), 7.84 (1H, d, J = 8.5), 8.27 (1H, s), 8.55 (1H, s), 8.81 (1H, s), 9.19 (1H, s).

Reference Example 55
2- (1- {6- [6- (pyrrol-1-yl) -indazol-1-yl] pyrazin-2-yl} -pyrrol-3-yl) acetic acid

56 mg of ethyl 2- (1- {6- [6- (pyrrol-1-yl) -indazol-1-yl] pyrazin-2-yl} -pyrrol-3-yl) acetate in 3 mL of methanol, 1,4-dioxane It melt | dissolved in 10 mL, 0.6 mL of 1M sodium hydroxide aqueous solution was added, and it stirred at room temperature for 2 hours. To the reaction solution was added 1M hydrochloric acid, and the mixture was extracted with ethyl acetate. The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated and 40 mg of 2- (1- {6- [6- (pyrrol-1-yl) -indazol-1-yl] Pyrazin-2-yl} -pyrrol-3-yl) acetic acid was obtained (yield 78%).
MS (ESI): 385 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 3.48 (2H, s), 6.31-6.40 (3H, m), 7.46-7.47 (2H, m), 7.65-7.68 (1H, m), 7.73 7.77 (2H, m), 8.02 (1H, d, J = 8.8), 8.57 (1H, s), 8.70 (1H, s), 8.93 (1H, s), 9.05 (1H, s), 12.30 (1H, s).

Reference Example 56
Ethyl 2- (1- {6- [4- (piperidin-1-yl) -indazol-1-yl] pyrazin-2-yl} -pyrrol-3-yl) acetate

Under argon atmosphere, trans-N, N'-dimethyl-1,2-cyclohexanediamine 8 mg, copper iodide 5 mg, 4-piperidin-1-yl-indazole 56 mg, tripotassium phosphate n-hydrate 178 mg, ethyl 2 100 mg of-[1- (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate was suspended in 1 mL of toluene and stirred at 110 ° C. for 24 hours. The reaction mixture was filtered through silica gel (ethyl acetate) and concentrated. The residue was purified by silica gel column chromatography (n-hexane / ethyl acetate = 10/1), and 110 mg of ethyl 2- (1- {6- [4- (piperidin-1-yl) -indazol-1-yl] Pyrazin-2-yl} -pyrrol-3-yl) acetate was obtained (91% yield).
MS (ESI): 431 (M + H) ^+
1 H-NMR (400 MHz, CDCl ₃ ) δ: 1.31 (3H, t, J = 7.1), 1.64-1.73 (2H, m), 1.79-1.90 (4H, m), 3.24-3.33 (4H, m), 3.60 (2H, s), 4.21 (2H, q, J = 7.1), 6.42-6.47 (1H, m), 6.72 (1H, d, J = 7.6), 7.45 (1H, t, J = 7.6), 7.48 -7.53 (1H, m), 7.55-7.58 (1H, m), 8.22 (1H, d, J = 8.3), 8.27 (1H, s), 8.50 (1H, s), 9.17 (1H, s).

Reference Example 57
2- (1- {6- [4- (piperidin-1-yl) -indazol-1-yl] pyrazin-2-yl} -pyrrol-3-yl) acetic acid

Dissolve 110 mg of ethyl 2- (1- {6- [4- (piperidin-1-yl) -indazol-1-yl] pyrazin-2-yl} -pyrrol-3-yl) acetate in 10 mL of 1,4-dioxane Then, 1.3 mL of 1M aqueous sodium hydroxide solution was added, and the mixture was stirred at room temperature for 2 hours. To the reaction solution was added 1M hydrochloric acid, and the mixture was extracted with ethyl acetate. The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and 92 mg of 2- (1- {6- [4- (piperidin-1-yl) -indazol-1-yl] Pyrazin-2-yl} -pyrrol-3-yl) acetic acid was obtained (90% yield).
MS (ESI): 403 (M + H) ^+
1 H-NMR (400 MHz, DMSO-d ₆ ) δ: 1.55-1.66 (2H, m), 1.67-1.82 (4H, m), 3.20-3.29 (4H, m), 3.50 (1H, s), 6.38 ( 1H, s), 6.75 (1H, d, J = 7.8), 7.50 (1H, t, J = 7.8), 7.72 (1H, s), 7.75 (1H, s), 8.14 (1H, d, J = 8.3 ), 8.53 (1H, s), 8.91 (1H, s), 9.05 (1H, s), 12.29 (1H, s).

Reference Example 58
Ethyl 2- {1- [6- (6-amino-1H-indazolyl) pyrazin-2-yl] pyrrol-3-yl} acetate

Under an argon atmosphere, trans-N, N′-dimethyl-1,2-cyclohexanediamine 16 mg, copper iodide 5 mg, 6-aminoindazole 89 mg, tripotassium phosphate · n hydrate 313 mg, ethyl-2- [1- 200 mg of (6-iodopyrazin-2-yl) pyrrol-3-yl] acetate was suspended in 1 mL of toluene and stirred at 110 ° C. for 24 hours. The reaction mixture was filtered through silica gel (ethyl acetate) and concentrated. The residue was purified by silica gel column chromatography (n-hexane / ethyl acetate = 4/6), and 105 mg of ethyl 2- {1- [6- (6-amino-1H-indazolyl) pyrazin-2-yl] pyrrole- 3-yl} acetate was obtained (52% yield).
MS (ESI): 363 (M + H) ^+
1 H-NMR (300 MHz, CDCl ₃ ) δ: 1.31 (3H, t, J = 7.1), 3.63 (2H, s), 4.22 (2H, q, J = 7.1), 6.40 (1H, brs), 6.72 ( 1H, dd, J = 2.0,8.5), 7.49 (1H, t, J = 2.0), 7.54 (1H, d, J = 8.5), 7.67 (1H, brs), 7.97 (1H, brs), 8.07 (1H , s), 8.45 (1H, s), 9.12 (1H, s).

Reference Example 59
2- {1- [6- (6-Amino-1H-indazolyl) pyrazin-2-yl] pyrrol-3-yl} acetic acid

50 mg of ethyl 2- {1- [6- (6-amino-1H-indazolyl) pyrazin-2-yl] pyrrol-3-yl} acetate is dissolved in 1 mL of methanol and 1 mL of 1,4-dioxane, and 1M sodium hydroxide 0.5 mL of an aqueous solution was added and stirred at room temperature for 2 hours. To the reaction solution was added 1M hydrochloric acid, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated and 40 mg of 2- {1- [6- (6-amino-1H-indazolyl) pyrazin-2-yl] pyrrole- 3-yl} acetic acid was obtained (87% yield).
MS (ESI): 335 (M + H) ^+
1 H-NMR (300 MHz, DMSO-d ₆ ) δ: 3.52 (2H, s), 5.90 (2H, brs), 6.37 (1H, brs), 6.70 (1H, d, J = 8.5), 7.53 (1H, d, J = 8.5), 7.68 (1H, s), 7.74 (1H, s), 7.81 (1H, t, J = 2.7), 8.21 (1H, s), 8.82 (1H, s), 8.99 (1H, s).

Reference Example 60-80
General synthesis of 2- (1- {6- [6- (alkylamino) -1H-indazol-1-yl] pyrazin-2-yl} -1H-pyrrol-3-yl) acetic acid compounds.

For synthesis of the compound, MiniBlock ^™ manufactured by Bohdan was used. 21 In all reaction vessels 2- {1- [6- (6-amino-1H-indazolyl) pyrazin-2-yl] pyrrol-3-yl} acetic acid (59 compounds of Reference Example) 15 mg, (Polystyrylmethyl) trimethylammonium borohydride ( Novabiochem, 4.1 mmol / g) 27 mg was added, 0.11 mmol of 21 aldehydes or ketones were added to each reaction vessel, suspended in 0.5 ml of 10% acetic acid-DMF solution, and stirred at 50 ° C. for 20 hours. All reaction mixtures were filtered and washed with 0.3 ml DMF. The filtrate was Agilent System 1100 (column: Waters Xterra ^™ Prep MS C ₁₈ 10 mm × 50 mm, developing phase: A = 0.1% formic acid-acetonitrile, B = 0.1% formic acid-water, gradient: A / B = 10/90. → 90/10 (6 minutes), flow rate: 4 ml / min) to obtain 21 target compounds (see Table 15).

2- (1- (6- (6- (Isopropylamino) -1H-indazol-1-yl) pyrazin-2-yl) -1H-pyrrol-3-yl) acetic acid (Reference Example 65 compound)
¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 1.27 (6H, d, J = 6.4), 3.47 (2H, s), 3.65 to 3.73 (1H, m), 6.29 (1H, d, J = 7.2 ), 6.40 (1H, brs), 6.74 (1H, d, J = 8.8), 7.54 (1H, d, J = 8.5), 7.65 (1H, s), 7.70 (1H, s), 7.77 (1H, brs) ), 8.22 (1H, s), 8.85 (1H, s), 9.02 (1H, s).
2- (1- (6- (6- (cyclopentylamino) -1H-indazol-1-yl) pyrazin-2-yl) -1H-pyrrol-3-yl) acetic acid (Compound of Reference Example 72)
¹ H-NMR (400 MHz, DMSO-d ₆ ) δ: 1.57-1.64 (4H, m), 1.70-1.75 (2H, m), 2.00-2.07 (2H, m), 3.47 (2H, s), 3.85 3.91 (1H, m), 6.39 (1H, brs), 6.43 (1H, d, J = 5.9), 6.76 (1H, d, J = 8.8), 7.54 (1H, d, J = 8.5), 7.64 (1H , s), 7.73 (1H, s), 7.78 (1H, brs), 8.23 (1H, s), 8.86 (1H, s), 9.03 (1H, s).

Example 1
Antiproliferative action of pyrazine derivatives in various cancer cells The above-mentioned Reference Examples 9, 31, 33, 47, 51, 61, 65, 72 and 73 compounds as test substances, leukemia-derived cell lines as cancer cell lines (Jurkat, L1210) ), Prostate cancer-derived cell line (PC-3), colon cancer-derived cell line (Colon26), lung cancer-derived cell line (PC10, LLC), breast cancer-derived cell line (SKBR3, Ehrlich) The effect of pyrazine derivatives on the growth inhibition of various cancer cells was examined. Culture media include Jurkat, L1210, PC-3, Colon26, PC10, LLC and Ehrlich for RPMI1640 (invitrogen) and SKBR3 for McCoy's 5A (invitrogen) with penicillin G at 100 U / mL, streptomycin at 0.1 mg / mL and fetal bovine Serum (FCS, high clone) supplemented to 10% was used. First, each cancer cell line was seeded on a 96-well flat bottom plate and cultured in a serum-free medium for 24-48 hours. The test substance was dissolved in dimethyl sulfoxide to prepare solutions of 50 mM, 10 mM and 2 mM. These solutions were diluted with a medium supplemented with 5% FCS to prepare solutions of 50 μM, 10 μM and 2 μM (dimethylsulfoxide final concentration 0.1%). This was dispensed on a plate and cultured under conditions of 5% CO ₂ and 37 ° C. for 24-48 hours. After completion of the culture, a WST-1 solution (Premix WST-1 Cell Proliferation Assay System, TaKaRa) was added and incubated at 37 ° C. for 30 minutes. After incubation, the absorbance at 450 nm based on the formazan dye produced by living cells was measured using a plate reader (BioRad). The percentage of cell growth inhibition (%) was calculated based on the following formula.

  Percentage of cell growth inhibition (%) = (absorbance of compound-added / serum-added well−absorbance of well without serum) / (absorbance of compound-free / serum-added well−absorbance of well without serum) × 100.

The IC ₅₀ value as an index of the cell growth inhibitory action was determined by preparing a graph plotting the logarithm of the test substance concentration and the cell growth inhibition percentage as the concentration at which the cell growth inhibition percentage was reduced by 50%.
As is apparent from Table 16, each test substance clearly inhibited the growth of various cancer cells of humans and mice and was shown to have an anticancer effect.

Example 2
Taking the sodium salt of the compound of Reference Example 72, which is a preferred example of a pyrazine derivative, the tumor growth inhibitory effect by administration of the anticancer agent of the present invention was examined using a mouse colon cancer model.

A tumor derived from Colon 26 cell, a mouse colon cancer cell line, was implanted subcutaneously in the back of a mouse (BALB / c strain, male, 6 weeks old, SLC Japan). As the tumor to be transplanted, a non-necrotic part extracted from a cancer-bearing mouse was cut into about 2 mm square. Cancer-bearing mice were prepared by intradermal injection of Colon26 cell suspension (4 × 10 ⁶ cells / 200 μL RPMI 1640) into BALB / c mice. Tumors were maintained by serial transplantation in BALB / c mice. On the 4th day after transplantation, body weight and tumor diameter were measured, and divided into a control group (n = 7) and a reference example 72 compound administration group (n = 7) based on the estimated tumor volume. The estimated tumor volume was calculated from the formula: 1/2 x tumor major axis x tumor minor axis ² . Reference Example 72 The compound is dissolved in physiological saline (Otsuka), and the dose is 20 mg / kg body weight, and is intravenously administered once a day from 0 to 3 days, starting on the day of grouping (day 0). Administered. Tumor diameter was measured on day 3 to calculate estimated tumor volume, and body weight was measured on day 4.

  The estimated tumor volume on the 3rd day after transplantation was suppressed by 29% in the reference example 72 compound-administered group compared with the control group (FIG. 1). In addition, regarding the body weight, there was no difference between the reference example 72 compound administration group and the control group (control group: 22.9 ± 0.5 g, reference example 72 compound administration group: 22.7 ± 0.5 g). From the above results, it was clarified that the compound of Reference Example 72 did not exhibit side effects even at the individual level, and suppressed tumor growth and exhibited anticancer activity.

It is a graph which shows the effect of the reference example 72 compound with respect to the estimated tumor volume in a mouse | mouth colon cancer model.

Claims (5)

Formula I

[Wherein, n represents an integer of 0-2, R ¹ represents a hydrogen atom or a C ₁ -C ₄ linear alkyl group, A represents an indolediyl group, a pyrrolediyl group, a furandiyl group, a thiophene group, Diyl group, pyrazole diyl group, isoxazole diyl group, isothiazole diyl group, imidazole diyl group, oxazole diyl group, thiazole diyl group or triazole diyl group, X is a nitrogen atom or CH, R ² and R ³ May be the same or different, and R ² and R ³ do not become hydrogen at the same time, C ₁ -C ₈ linear alkyl group, C ₃ -C ₈ branched alkyl group, C _3- A cyclic alkyl group of C ₈ (these alkyl groups may be substituted by a hydroxyl group, an amino group, a saturated or unsaturated 5- or 6-membered heterocyclic group), -C (O) R ⁴ , -SO It represents ₂ R ^4, R ⁴ is hydrogen, straight-chain alkyl group of C ₁ -C _3, C Branched alkyl group having ₃ -C _5, represents a phenyl group. R ² and R ³ may include a nitrogen atom to which they are bonded to form a saturated or unsaturated 5- or 6-membered heterocyclic group, and this heterocyclic group is one or more nitrogen, oxygen And may further contain a sulfur atom. In this heterocyclic group, one or two hydrogen atoms of a C ₁ -C ₄ alkyl group, a hydroxyl group, a C ₁ -C ₄ alkoxy group, an amino group, or an amino group which may be substituted with a hydroxyl group are each independently C ₁ linear alkyl group -C _3, may be substituted by an amino group which is substituted by branched alkyl groups of C ₃ -C _5. The nitrogen atom to which R ² and R ³ are bonded may be bonded to any of the 3, 4, 5, 6 and 7 positions of Formula I. ]
The anticancer agent which uses the pyrazine derivative represented by these, or its pharmaceutically acceptable salt as an active ingredient.   2. The anticancer agent comprising a pyrazine derivative or a pharmaceutically acceptable salt thereof according to claim 1, wherein n is 0 or 1 in formula I.   An anticancer agent comprising the pyrazine derivative or a pharmaceutically acceptable salt thereof as an active ingredient according to claim 2, wherein A is an indolediyl group, a pyrrolediyl group, a furandiyl group, or a pyrazolediyl group.   In formula I, X is a nitrogen atom, The anticancer agent which uses the pyrazine derivative or its pharmaceutically acceptable salt of Claim 3 as an active ingredient.   The anticancer comprising the pyrazine derivative or the pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein the cancer is colon cancer, lung cancer, prostate cancer, liver cancer, breast cancer or leukemia. Agent.

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