JP2014034533A - Combination of hsp90 inhibitor and egfr tyrosine kinase inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a combination therapy of an HSP90 inhibitor targeting HSP90 greatly involved in survival of cancer cells and other antitumor agents.SOLUTION: An antitumor agent is obtained by combining a triazole compound (A) represented by general formula (1) with an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (B).

Description

  The present invention relates to an antitumor agent comprising a combination of a triazole compound having HSP90 inhibitory activity and an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor. In addition, an antitumor effect potentiator of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, and epidermal growth factor receptor (EGFR) tyrosine kinase inhibition, characterized by comprising a triazole compound having the HSP90 inhibitory activity The present invention relates to a method for enhancing the antitumor effect of an agent.

  Development of new antitumor chemotherapy is required for the treatment of malignant tumor diseases. In recent years, various functional molecules such as growth factors and growth factor receptors related to signal transduction involved in cell proliferation, and proteins related to signal transduction pathways have been recognized. Thus, molecular target antitumor agents have been developed that target these functional molecules and suppress their functions.

Heat shock protein (HSP) is a molecular chaperone present in cells, and is a functional molecule classified into several families such as HSP90, HSP70, HSP60, HSP40, and small HSPs depending on the molecular weight. Molecular chaperone is a general term for proteins that temporarily form a complex with a target protein in order to promote the formation of a functional higher-order structure of the protein. That is, molecular chaperones have the activity of helping protein folding and association and inhibiting aggregation.
HSP90 is an abundant molecular chaperone that accounts for 1 to 2% of the total soluble protein in the cell, is uniformly distributed in the cytoplasm, and exists mainly as a dimer. HSP90 is responsible for the function of folding proteins that are not in a denatured or folded state, depending on ATP. HSP90 is known to interact with various proteins involved in intracellular signal transduction systems. That is, HSP90 is often required for functional expression of various target proteins, and the mechanism of action is that HSP90 specifically recognizes a protein in an unstable folded state and binds to this to form a complex. Based on biochemical properties that form. The activity of HSP90 alone in protein folding is weak and functions in cooperation with other molecular chaperones (cochaperones) such as HSP70 and p23 having the same folding activity.
Various target proteins (steroid receptors, Raf serine kinases, tyrosine kinases) involved in cancer-related signal transduction depend on HSP90 for structural construction. Therefore, it has been clarified that HSP90 is deeply involved in cell cycle control and cell canceration / proliferation / survival signals.

The regulation of many signal molecules is lost in human tumors, and tumors require HSP90 in order to maintain the function of these signal molecules (Non-patent Document 1). Therefore, compounds that act on HSP90 and inhibit its function are known to cause degradation of various target proteins involved in cancer-related signal transduction and are known to suppress the growth of cancer cells based on it. ing. That is, the HSP90 inhibitor changes the structure of the chaperone complex including the target protein and HSP90, and degrades the target protein released from the complex mainly in the ubiquitin / proteasome system. As a result, the target protein amount of HSP90 is reduced, blocking downstream signal transmission associated therewith and suppressing the growth of cancer cells, thereby providing an antitumor effect.
In particular, in the process of becoming cancerous, a plurality of gene abnormalities are accumulated and protein mutations occur. That is, in cancer cells, many mutant proteins are produced, and more chaperone activity is required than normal cells consisting of normal proteins. For this reason, it is known that the expression level of HSP90 is increasing in many cancer cells. From these facts, HSP90 inhibitors are expected to act selectively on cancer cells rather than normal cells. Furthermore, in addition to the fact that abnormal protein expression is observed, cancer cells are placed in a hypoxic state and a nutrient starvation state, and are under a kind of stress state. Therefore, the dependence on chaperone activity by HSP90 may be high. Conceivable. Therefore, cancer cells are expected to be more sensitive to HSP90 inhibitors than normal cells. Therefore, exploratory studies of HSP90 inhibitors targeting HSP90 as well as verification of their antitumor effects have been made.

  As an exploratory study of HSP90 inhibitors, Patent Document 1 discloses that 5- (2,4-dihydroxyphenyl)-[1,2,4] triazol-3-one derivatives have HSP90 inhibitory activity and cancer cells. It describes that it is a compound having growth inhibitory activity. It has been reported that the compound shows an excellent antitumor effect even in animal experiments and is promising as an anticancer agent. Patent Document 2 or Patent Document 3 also reports triazole compounds having HSP90 inhibitory activity.

The epidermal growth factor receptor (EGFR (erbB1 or HER-1)) belonging to the human epidermal growth factor receptor family is composed of an extracellular ligand binding region, a transmembrane region, and an intracellular tyrosine kinase region. It is a transmembrane receptor having tyrosine kinase activity and is a type I transmembrane glycoprotein having a molecular weight of 170 KDa. EGFR shows homology with other erbB family proteins erbB2 (HER2 / neu), erbB3 (HER3), erbB4 (HER4) in its structure, and EGFR is homo- or heterodimer (dimer) with erbB family proteins. Form. When a ligand such as epidermal growth factor (EGF) or transforming growth factor-α (TGF-α) binds to the EGFR extracellular ligand binding region, EGFR forms a dimer, and each receptor molecule The upper tyrosine residues are autophosphorylated with each other. Subsequent to this autophosphorylation, a downstream intracellular signal transduction system is activated, a signal for cell proliferation is transmitted to the nucleus, and a series of events that stimulate cell proliferation occurs.
EGFR is expressed in normal tissues such as epithelial tissues and mesenchymal tissues, and plays a major role in processes such as cell proliferation, regeneration, differentiation, and development. On the other hand, the expression level of EGFR and the frequency of expression thereof are higher in tumor cells than in normal cells, such as head and neck cancer, breast cancer, lung cancer, colon cancer, prostate cancer, kidney cancer, uterine cancer, ovarian cancer, and bladder cancer. Overexpression in many solid tumors has been reported, and EGFR overexpression is thought to be involved in tumor progression, prognosis, and chemotherapy resistance. Therefore, receptor protein molecules involved in signal transduction including EGFR are considered as one of new target molecules in cancer treatment. In particular, the activation of EGFR tyrosine kinase that triggers signal transmission from EGFR is considered to be involved in tumor growth such as cell cycle enhancement, apoptosis inhibition, invasion, metastasis, and angiogenesis in tumors.
Erlotinib (Tarceva (registered trademark)), lapatinib (Tykerb (registered trademark)), gefitinib (Iressa (registered trademark)), etc. are known as antitumor agents whose main mechanism of action is EGFR tyrosine kinase inhibitory action . These suppress EGFR autophosphorylation by inhibiting EGFR tyrosine kinase, and inhibit cell division by inhibiting cell proliferation signal transduction caused by binding of a ligand to EGFR. Therefore, the cell division of the tumor cell which expresses EGFR is inhibited, and the effect which suppresses tumor growth is shown.

For malignant tumor diseases, the degree of medical satisfaction with pharmacotherapy is not sufficient, and combination chemotherapy using a combination of a plurality of drugs is performed for the purpose of enhancing the therapeutic effect. In other words, combination chemotherapy combining a plurality of drugs with different mechanisms of action and side effects has been developed, which contributes to improvement of treatment results. In particular, in recent years, molecular target antitumor agents that target molecules involved in signal transduction involved in cell proliferation have been developed and put into clinical use. Therefore, there is a demand for the establishment of a combination therapy having a higher therapeutic effect using these molecular target antitumor agents.
Even in the case of HSP90 inhibitors, combination therapy with other antitumor agents has been attempted for the purpose of improving antitumor effects and improving sensitivity to drug-resistant tumors (Non-patent Document 2).
The HSP90 inhibitor exerts a cancer cell growth inhibitory action by inhibiting the complex forming function of HSP90 and the target protein. Therefore, the combined use of an HSP90 inhibitor and an antitumor agent whose action mechanism is the functional inhibition of the target protein (client protein) is expected to enhance the antitumor effect. Since EGFR is known as a client protein of HSP90, combination therapy of an EGFR tyrosine kinase inhibitor that inhibits a signal transduction function from EGFR and an HSP90 inhibitor is being studied. Patent Document 4 describes the combined use of a triazole compound having HSP90 inhibitory activity and erlotinib. Non-Patent Document 3 describes the combined use of IPI-504, known as an HSP90 inhibitor, and lapatinib. Furthermore, Non-Patent Document 4 describes the combined use of 17-AAG known as an HSP90 inhibitor and gefitinib.

Hsp90 inhibitors as novel cancer chemotherapeutic agents. Trends in Molecular Medicine. 2002, 8: 4 (Suppl.), S55-61. Hsp90 inhibitors and drug resistance in cancer: the potential benefits of combina- tion therapies of Hsp90 inhibitors and other anti-cancer. Biochemical Pharmacology 2012, 83: 8, 995-1004. Antitumor efficacy of IPI-504, a selective heat shock protein 90 inhibitor against human epidermal growth factor receptor 2-positive human xenograft models as a single agent and in combination with trastuzumab or lapatinib. Molecular Cancer Therapeutics, 2009, 8, 2131-2141 Cooperative inhibitory effect of ZD1839 (Iressa) in combination with 17-AAG on glioma cell growth. Molecular Carcinogenesis, 2006, 45, 288-301

International Publication WO2006 / 095783 International Publication No. WO2009 / 023211 International Publication WO2007 / 134678 International publication WO2011 / 133520

Since HSP90 depends on the survival of cancer cells, development of antitumor agents using HSP90 as a target molecule has been underway. An object of the present invention is to provide a combination therapy with another antitumor agent as a drug therapy having a higher therapeutic effect using this HSP90 inhibitor. That is, to provide a combination therapy that exhibits an excellent therapeutic effect by using 5- (2,4-dihydroxyphenyl)-[1,2,4] triazol-3-one derivatives and other antitumor agents as HSP90 inhibitors. Is an issue.
Further, according to another viewpoint, 5- (2,4-dihydroxyphenyl)-[1,2,4] triazol-3-one derivatives exert the therapeutic effect enhancing action of existing antitumor agents. The purpose is to provide an antitumor agent as an antitumor effect enhancer.
In yet another aspect, an object of the present invention is to provide a combination therapy that further enhances the therapeutic effect in EGFR tyrosine kinase inhibitor therapy for the treatment of malignant tumors.

［式中、Ｘは直鎖状または分岐状の炭素数１〜８のアルキル基、炭素数２〜１０のアルキニル基、またはハロゲン原子を示し、Ｙは硫黄原子または酸素原子を示し、ｍは０〜４の整数を示し、Ａは置換基を有するアミノ基を示す］で表されるトリアゾール化合物（Ａ）と、上皮増殖因子受容体（ＥＧＦＲ）チロシンキナーゼ阻害剤（Ｂ）を組み合わせた抗腫瘍剤。 As a result of intensive studies, the present inventors combined a 5- (2,4-dihydroxyphenyl)-[1,2,4] triazol-3-one derivative represented by the general formula (1) with an EGFR tyrosine kinase inhibitor. However, it has been found that the antitumor effect is remarkably superior as compared with the antitumor effect of each single use. That is, the gist of the present invention is as follows.
(1) The following general formula (1)

[Wherein, X represents a linear or branched alkyl group having 1 to 8 carbon atoms, an alkynyl group having 2 to 10 carbon atoms, or a halogen atom, Y represents a sulfur atom or an oxygen atom, and m represents 0 An antitumor agent comprising a combination of a triazole compound (A) represented by the following formula: and an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (B) .

［式中、Ｘは直鎖状または分岐状の炭素数１〜８のアルキル基、炭素数２〜１０のアルキニル基、またはハロゲン原子を示し、Ｙは硫黄原子または酸素原子を示し、ｍは０〜４の整数を示し、Ａは置換基を有するアミノ基を示す］で表されるトリアゾール化合物（Ａ）を有効成分として含有する事を特徴とする、上皮増殖因子受容体（ＥＧＦＲ）チロシンキナーゼ阻害剤の抗腫瘍効果を増強させる抗腫瘍効果増強剤。 (2) According to another viewpoint, the following general formula (1)

[Wherein, X represents a linear or branched alkyl group having 1 to 8 carbon atoms, an alkynyl group having 2 to 10 carbon atoms, or a halogen atom, Y represents a sulfur atom or an oxygen atom, and m represents 0 Inhibition of epidermal growth factor receptor (EGFR) tyrosine kinase, comprising as an active ingredient a triazole compound (A) represented by an integer of ˜4, A represents an amino group having a substituent]] An antitumor effect potentiator that enhances the antitumor effect of the agent.

［式中、Ｘは直鎖状または分岐状の炭素数１〜８のアルキル基、炭素数２〜１０のアルキニル基、またはハロゲン原子を示し、Ｙは硫黄原子または酸素原子を示し、ｍは０〜４の整数を示し、Ａは置換基を有するアミノ基を示す］で表されるトリアゾール化合物（Ａ）を併用で投与する事を特徴とする、上皮増殖因子受容体（ＥＧＦＲ）チロシンキナーゼ阻害剤の抗腫瘍効果を増強させる方法、を包含する。 (3) In the administration of an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor in cancer therapy, the following general formula (1)

[Wherein, X represents a linear or branched alkyl group having 1 to 8 carbon atoms, an alkynyl group having 2 to 10 carbon atoms, or a halogen atom, Y represents a sulfur atom or an oxygen atom, and m represents 0 An epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, characterized by administering a triazole compound (A) represented by the formula: A method of enhancing the antitumor effect of

According to the present invention, an antitumor improved in therapeutic effect by using a combination of a 5- (2,4-dihydroxyphenyl)-[1,2,4] triazol-3-one derivative and an EGFR tyrosine kinase inhibitor. Drug therapy can be provided. By applying the triazole derivative to the EGFR tyrosine kinase inhibitor therapy conventionally used in clinical treatment, the antitumor effect of the EGFR tyrosine kinase inhibitor can be enhanced and the therapeutic effect can be improved.
Accordingly, it is possible to achieve further improvement of the antitumor effect by the conventional method, and to sufficiently suppress the growth of malignant tumors with reduced sensitivity to the EGFR tyrosine kinase inhibitor and maintain the therapeutic effect.
Furthermore, in the case of obtaining an antitumor effect equivalent to the use of an antitumor agent alone, the dose of each antitumor agent can be relatively reduced, and the occurrence of undesirable pharmacological effects is adversely affected by the antitumor effect. It can be reduced without affecting.

Erlotinib (B-1), triazole compound (A-1), gemcitabine (C-1) and comparative HSP90 inhibitor (D) monotherapy using mice transplanted subcutaneously with human pancreatic cancer cells BxPC3, and erlotinib (B-1) and triazole compound (A-1) combination therapy, erlotinib (B-1) and comparative HSP90 inhibitor (D) combination therapy, and erlotinib (B-1) and triazole compound (A-1 ) And gemcitabine (C-1) in combination with antitumor effect.

The present invention is characterized in that the triazole compound (A) represented by the general formula (1) is used in combination with an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (B). Furthermore, it is characterized in that gemcitabine is arbitrarily used in combination. The details will be described below.
In the present invention, the triazole compound (A) having HSP90 inhibitory activity is a triazole compound represented by the general formula (1). That is, in the triazole ring, the substitution position of the Y group is the 3-position, the substitution position of the phenyl group having the A group is the 4-position, and the substitution position of the 2,4-dihydroxyphenyl group having the X group is the 5-position It is a [1,2,4] triazole derivative.
The triazole compound represented by the general formula (1) has a keto-enol tautomer and can have a keto isomer structure represented by the following general formula (1K). That is, in the present invention, (1) and (1K) are the same compound, and the triazole compound (A) according to the present invention includes the compound represented by the general formula (1K).

X in the general formula (1) is a linear or branched alkyl group having 1 to 8 carbon atoms which may have a substituent, or an alkynyl having 2 to 10 carbon atoms which may have a substituent. A group, or a halogen atom. X will be described below.
Examples of X in the general formula (1) include a linear or branched alkyl group having 1 to 8 carbon atoms. The alkyl group is preferably a methyl group, ethyl group, n-propyl group, isopropyl group, cyclopropyl group, n-butyl group, sec-butyl group, tert-butyl group, 2,2-dimethylpropyl group, n- Examples include a hexyl group, a cyclohexyl group, an n-heptyl group, a cyclohexylmethyl group, an n-octyl group, a cyclohexylethyl group, and the like.
The alkyl group may have a substituent. As the substituent in the case of having a substituent, for example, a mercapto group, a hydroxyl group, a halogen atom, a nitro group, a cyano group, a carbocyclic or heterocyclic aryl group, a C1-C4 alkylthio group, an arylthio group, a C1-C1 4-alkoxy group, aryloxy group, aliphatic or aromatic amino group, acyl group, carboxyl group and the like can be mentioned. Examples of the alkyl group having a substituent as X include 2,2,2-trichloroethyl group, trifluoromethyl group, pentafluoroethyl group, trifluoromethyl-2,2,2-trifluoroethyl group, N , N-dimethylaminomethyl group, N, N-dimethylaminoethyl group, morpholinylmethyl group, piperidinylmethyl group, hydroxymethyl group, 1-hydroxyethyl group, 2-hydroxyethyl group, 1-hydroxy-1 -Methyl-ethyl group, methoxyethyl group, methoxymethyl group, benzyl group, 2-phenylethyl group, pyridylmethyl group and the like can be mentioned.
As the linear or branched alkyl group having 1 to 8 carbon atoms in X, an ethyl group, an isopropyl group, a tert-butyl group, and a 2,2-dimethylpropyl group are preferable, and an isopropyl group is particularly preferable.

  Examples of X in the general formula (1) include alkynyl groups having 2 to 10 carbon atoms. The alkynyl group is a 1-alkynyl group or a 2-alkynyl group. The alkynyl group may have a substituent. As the substituent in the case of having a substituent, a halogen atom, a carbocyclic or heterocyclic aryl group, a C1-C4 alkyloxy group, a C1-C4 alkylthio group, an aliphatic or aromatic amino group, an acyl group, And alkylsilyl groups. Preferred 1-alkynyl groups include ethynyl group, 3,3-dimethylbut-1-ynyl group, 2-phenyl-ethynyl group, 2-trimethylsilyl-1-ethynyl group, and the like. Examples of the 2-alkynyl group include prop-2-ynyl group, but-2-ynyl group, 3-phenylprop-2-ynyl group, 4,4-dimethylpent-2-ynyl group, and 3-trimethylsilylpropa-2. -Inyl group, etc. are mentioned. A 2-alkynyl group having 2 to 10 carbon atoms is preferable, and a prop-2-ynyl group or a but-2-ynyl group is particularly preferable.

  Examples of X in the general formula (1) include a halogen atom. As a halogen atom, a fluorine atom, a chlorine atom, a bromine atom, or an iodine atom is shown. As the halogen atom, a bromine atom is preferable.

  X in the general formula (1) is preferably a linear or branched alkyl group having 1 to 8 carbon atoms or a 2-alkynyl group having 2 to 10 carbon atoms. The substituent X is particularly preferably an ethyl group, an isopropyl group, a tert-butyl group, a prop-2-ynyl group, or a but-2-ynyl group.

In the general formula (1), A represents an amino group having a substituent. Examples of the amino group having a substituent in A include an aliphatic primary amino group and an aliphatic secondary amino group.
Examples of the aliphatic primary amino group include N-alkylamino groups having a linear, branched or cyclic alkyl group having 1 to 8 carbon atoms. For example, a methylamino group, isopropylamino group, n-butylamino group, cyclohexylamino group, and the like can be mentioned. Examples of the aliphatic secondary amino group include an N, N-dialkylamino group having a linear, branched or cyclic alkyl group having 1 to 8 carbon atoms, or a cyclic secondary amino group. Examples of the N, N-dialkylamino group include dimethylamino group, diisopropylamino group, N-methyl-N-butylamino group, N-methyl-N-cyclohexylamino group, N, N-dicyclohexylamino group, and the like. It is done. Examples of the aliphatic cyclic secondary amino group include a morpholino group, a 4-methylpiperazin-1-yl group, a piperidin-1-yl group, and a pyrrolidin-1-yl group. The aliphatic amino group represented by A is preferably an aliphatic cyclic secondary amino group, and is a morpholino group, 4-methylpiperazin-1-yl group, piperidin-1-yl group, pyrrolidin-1-yl group. Is preferred. Of these, a morpholino group or a 4-methylpiperazin-1-yl group is particularly preferable.
In general formula (1), m represents an integer of 0 to 4. Among these, m is preferably 0 or 1.
That is, the — (CH ₂ ) _m —A group in the general formula (1) is preferably a cyclic secondary amino group in which m is 0 or 1 and A is an aliphatic group, a morpholino group, a methyl-morpholino group, 4 -Methylpiperazin-1-yl group, methyl-4-methylpiperazin-1-yl group, piperidin-1-yl group, methyl-piperidin-1-yl group, pyrrolidin-1-yl group, methyl-pyrrolidin-1- An yl group is particularly preferred.

Particularly preferred examples of the triazole compound (A) in the present invention are given below.
5- (2,4-Dihydroxy-5-isopropyl-phenyl) -4- [4- (4-methyl-piperazin-1-ylmethyl) -phenyl] -2,4-dihydro- [1,2,4] triazole -3-one; (A-1),
4-Isopropyl-6- {5-mercapto-4- [4- (morpholin-4-yl) -phenyl] -4H- [1,2,4] triazol-3-yl} -benzene-1,3-diol ; (A-2),
4-Isopropyl-6- {5-mercapto-4- [4- (morpholin-4-ylmethyl) -phenyl] -4H- [1,2,4] triazol-3-yl} -benzene-1,3-diol ; (A-3),
4- {5-hydroxy-4- [4- (morpholin-4-yl) -phenyl] -4H- [1,2,4] triazol-3-yl} -6-isopropyl-benzene-1,3-diol ; (A-4),
4- {5-hydroxy-4- [4- (morpholin-4-ylmethyl) -phenyl] -4H- [1,2,4] triazol-3-yl} -6-isopropyl-benzene-1,3-diol ; (A-5),
5- [5- (Butyn-2-yl) -2,4-dihydroxy-phenyl] -4- [4- (morpholin-4-ylmethyl) -phenyl] -2,4-dihydro- [1,2,4 ] Triazol-3-one; (A-6),
4- (Butyn-2-yl) -6- {5-mercapto-4- [4- (morpholin-4-ylmethyl) -phenyl] -4H- [1,2,4] triazol-3-yl] -benzene -1,3-diol; (A-7),
4-Bromo-6- {5-mercapto-4- [4- (morpholin-4-yl) -phenyl] -4H- [1,2,4] triazol-3-yl} -benzene-1,3-diol ; (A-8),
5- [2,4-Dihydroxy-5- (propyn-2-nyl) -phenyl] -4- [4- (morpholin-4-ylmethyl) -phenyl] -2,4-dihydro- [1,2,4 ] Triazol-3-one; (A-9)

  The triazole compound (A) according to the general formula (1) of the present invention can be produced by a known production method. For example, it can be synthesized according to the production method disclosed in International Publication WO2006 / 095783. For example, the above 5- (2,4-dihydroxy-5-isopropyl-phenyl) -4- [4- (4-methyl-piperazin-1-ylmethyl) -phenyl] -2,4-dihydro- [1,2, 4] Triazol-3-one; (A-1) has a production method disclosed in Example 2-5 of the patent document, and can be synthesized according to the method described therein. Other derivatives can also be prepared according to the methods disclosed in the aforementioned prior art documents.

The triazole compound (A) of the present invention may form a salt with an acid or a base, and a salt of the compound represented by the general formula (1) may be used. Examples of the salt with an acid include inorganic acid salts such as hydrochloride, hydrobromide and sulfate, and salts with organic acids such as trifluoroacetic acid, methanesulfonic acid and p-toluenesulfonic acid. . As a salt with a base, a sodium salt etc. can be mentioned, for example. These salts can be produced by a conventional method.
The triazole compound (A) of the present invention may be prepared by using a pharmacologically active ingredient as it is, and preparing commonly used preparations such as injections, instillations, tablets, capsules, powders etc. together with preparation bases. You can use it. In formulating, pharmaceutically acceptable carriers usually used as formulation bases, such as binders, lubricants, disintegrants, solvents, excipients, solubilizers, dispersants, stabilizers, suspensions. Turbidizing agents, preservatives, soothing agents, pigments, and fragrances can be used. The triazole compound (A) is formulated into the above-mentioned pharmaceutical preparation and can be administered by a known administration route such as intravenous administration, transarterial administration, subcutaneous administration, oral administration, transmucosal administration and the like.

Next, the epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (B) in the present invention will be described. The EGFR tyrosine kinase inhibitor used in the present invention suppresses EGFR autophosphorylation using EGFR as a target molecule, and inhibits cell proliferation signal transduction caused by binding of a ligand to EGFR, thereby expressing a tumor expressing EGFR Represents an antitumor agent whose main action is to suppress cell growth. In the present invention, the EGFR tyrosine kinase inhibitor is not particularly limited as long as it is a compound that suppresses EGFR phosphorylation. At present, as EGFR tyrosine kinase inhibitors, erlotinib, lapatinib, gefitinib, afatinib (BIBW2992), neratinib (HKI-272), etc. are known, and these are preferred EGFR tyrosine kinase inhibitors (B) according to the present invention. Can be listed.
The EGFR tyrosine kinase inhibitor (B) may be an pharmacologically active ingredient such as erlotib, lapatinib, gefitinib, afatinib, or neratinib as described above, or a free base or other salt that is acceptable as a pharmaceutical. Further, it may be a prodrug that releases these pharmacologically active ingredients in vivo.
As the EGFR tyrosine kinase inhibitor (B) in the present invention, erlotinib, lapatinib and gefitinib are particularly preferable, and erlotinib / hydrochloride, lapatinib tosylate and gefitinib (free base) are preferably used. These EGFR tyrosine kinase inhibitors include erlotinib hydrochloride (Tarceva®), lapatinib tosylate (Tykerb®), gefitinib (Iressa®) and pharmaceutical formulations with excipients, respectively. Therefore, the EGFR tyrosine kinase inhibitor (B) according to the present invention is preferably used as it is, and is preferably administered orally.

In the present invention, the anti-tumor agent in which the triazole compound (A) having an HSP90 inhibitory action and the EGFR tyrosine kinase inhibitor (B) are combined includes the triazole compound (A) and the EGFR tyrosine kinase inhibitor ( B) represents an anti-tumor agent comprising a mode of use administered to one patient. That is, the triazole compound (A) and the EGFR tyrosine kinase inhibitor (B) are intended to be used in combination for a malignant tumor disease patient. That is, it refers to an antitumor agent comprising separately administering the triazole compound (A) and the EGFR tyrosine kinase inhibitor (B) to one patient simultaneously, sequentially, or before and after. Therefore, the aspect of the antitumor agent containing the triazole compound (A) and the EGFR tyrosine kinase inhibitor (B) is not particularly limited, and any form may be used as long as these can be administered in combination. For example, the triazole compound (A) and the EGFR tyrosine kinase inhibitor (B) may be separate preparations. Further, it may be a pharmaceutical preparation prepared by mixing the triazole compound (A) and the EGFR tyrosine kinase inhibitor (B). In another embodiment, the kit may include a preparation type containing the triazole compound (A) and a preparation type containing the EGFR tyrosine kinase inhibitor (B), which are intended to be used in combination. Or the aspect of the kit containing the formulation type | mold containing this triazole compound (A), the formulation type | mold containing this EGFR tyrosine kinase inhibitor (B), and the container containing the said 2 formulation types can be mentioned.
In the present invention, the triazole compound (A) and the EGFR tyrosine kinase inhibitor (B) may be used at the same time, and after the triazole compound (A) is administered, the EGFR tyrosine kinase inhibitor (B) is used. It may be administered or vice versa. The triazole compound (A) may be a single dose or an intermittent dose, and the EGFR tyrosine kinase inhibitor (B) may be a combination of daily doses. The administration of the triazole compound (A) and the EGFR tyrosine kinase inhibitor (B) is a common administration method acceptable for pharmaceuticals such as single administration, intermittent administration, and daily administration for each drug. Is included. For example, the triazole compound (A) may be administered once and the EGFR tyrosine kinase inhibitor (B) may be administered every day. The triazole compound (A) can be administered by intravenous infusion over a relatively short time. The EGFR tyrosine kinase inhibitor (B) may be used by oral administration every day.

In the present invention, the respective doses of the triazole compound (A) and the EGFR tyrosine kinase inhibitor (B) should be determined while confirming the antitumor effect and side effects by an appropriate clinical test. Preferably, the triazole compound (A) is appropriately administered in clinical trials in which the triazole compound (A) is appropriately increased in a setting for daily or complete administration at an optimal acceptable dose. The respective doses of A) and the EGFR tyrosine kinase inhibitor (B) can be set. However, since the present invention can achieve the enhancement of the antitumor effect of the EGFR tyrosine kinase inhibitor, it is possible to reduce the amount of the EGFR tyrosine kinase inhibitor. Therefore, the optimal dose of the EGFR tyrosine kinase inhibitor should be increased or decreased as appropriate. These dose setting methods are test methods obvious to those skilled in the art, and can be directly determined based on the results based on appropriate clinical test methods.
In addition, based on the results of basic antitumor tests using animals, it is preferable to use 0.1 to 1000 mg of the triazole compound (A) per dose. On the other hand, the dose per administration of the EGFR tyrosine kinase inhibitor (B) is preferably 1 to 5000 mg based on known pharmacological activity results, and these can be used in appropriate combination. In particular, when erlotinib is used as the EGFR tyrosine kinase inhibitor (B), the dose per administration is 50 to 300 mg, which is preferably administered every day. Incidentally, for clinical use of erlotinib, it is recommended that 100 mg or 150 mg of erlotinib hydrochloride is orally administered to adults once a day. Moreover, when using lapatinib, the dose per time is 500-3000 mg, and it is preferable to administer every day. In general, lapatinib is orally administered to adults as 1250 mg of lapatinib tosylate once a day, and is used in combination with capecitabine in the treatment of breast cancer. Moreover, when using gefitinib, the dosage per time is 100-500 mg, and it is preferable to administer every day. The recommended dosage regimen for gefitinib is to use 250 mg of gefitinib once daily by oral administration to adults. In the present invention, the EGFR tyrosine kinase inhibitor (B) is preferably used at each clinically recommended dose, and is preferably used by appropriately increasing or decreasing the dose based on the recommended dose.

  The antitumor agent of the present invention is used for the treatment of malignant tumor diseases. The malignant tumor applied to the treatment according to the present invention is not particularly limited, and breast cancer, non-small cell lung cancer, small cell lung cancer, colorectal cancer, non-Hodgkin lymphoma (NHL), renal cell cancer, prostate cancer, liver Cellular cancer, gastric cancer, pancreatic cancer, soft tissue sarcoma, Kaposi sarcoma, carcinoid carcinoma, head and neck cancer, melanoma, ovarian cancer, cholangiocarcinoma, mesothelioma, and multiple myeloma Can be applied to. In particular, it is suitable for the treatment of malignant tumors in which abnormal expression or activation of EGFR contributes as a growth factor. That is, it is a cancer type in which EGFR overexpression or mutant EGFR overexpression is observed, such as breast cancer, non-small cell lung cancer, small cell cancer, bladder cancer, brain tumor, head and neck cancer, gastric cancer, pancreatic cancer, hepatocellular carcinoma, Suitable for treatment of bile duct cancer, colorectal cancer, mesothelioma, and multiple myeloma. In particular, EGFR overexpression and mutation have been observed in some squamous cell carcinomas, breast cancers, and brain tumors, and the antitumor of the present invention is useful in the treatment of brain tumors such as breast cancer, non-small cell lung cancer, malignant glioma, and glioblastoma. Agents can be applied. It can also be applied to breast cancer and non-small cell lung cancer with reduced sensitivity to EGFR tyrosine kinase inhibitors such as erlotinib, lapatinib, and gefitinib.

  The antitumor agent obtained by combining the triazole compound (A) and the EGFR tyrosine kinase inhibitor (B) of the present invention can be used in combination with another antitumor agent (C). Other antitumor agents (C) are not particularly limited, and pharmaceuticals approved as antitumor agents can be used. That is, alkylating agents such as cyclophosphamide, ifosfamide, mitomycin C, platinum complexes such as cisplatin, carboplatin, ogisaliplatin, anthracycline antitumor agents such as doxorubicin, epirubicin, pirarubicin, amrubicin, etoposide, etoposide phosphate, teniposide, etc. Etoposides, camptothecins such as irinotecan and nogitecan, taxanes such as paclitaxel and docetaxel, vinca alkaloids such as vincristine, vinblastine, vindesine and vinorelbine, 5-fluorouracil, tegafur, tegafur and uracil combination (UFT), Nucleic acid charges such as gimeracil / oteracil calcium combination (S-1), flutulon, capecitabine, gemcitabine, cytosine arabinoside Antagonists, methotrexate, folate antimetabolites such as pemetrexed, prednisolone, steroids such as dexamethasone, can be used trastuzumab, anti-EGFR antibody cetuximab such, the anti-VEGF antibody bevacizumab like. In particular, a combination of drugs that have been confirmed to have a combined effect with an EGFR tyrosine kinase inhibitor is preferable, and a combination with at least one selected from capecitabine, gemcitabine, and trastuzumab as the other antitumor agent (C) is preferable.

The combination example of the especially preferable triazole compound (A) of this invention and an EGFR tyrosine kinase inhibitor (B) is given.
The triazole compound (A) is 5- (2,4-dihydroxy-5-isopropyl-phenyl) -4- [4- (4-methyl-piperazin-1-ylmethyl) -phenyl] -2,4-dihydro- [1,2,4] Triazol-3-one; (A-1) is preferably used.
A combination of the above (A-1) and erlotinib (B-1) is preferable, and effective in the treatment of non-small cell lung cancer and pancreatic cancer. That is, it is possible to provide a method for treating non-small cell lung cancer or pancreatic cancer including administering each effective dose of (A-1) and erlotinib (B-1). Furthermore, an antitumor agent obtained by further combining gemcitabine (C-1) with (A-1) and erlotinib (B-1) is particularly preferable because it exhibits a remarkable antitumor effect. By administering the effective doses of (A-1), erlotinib (B-1) and gemcitabine (C-1), an effective treatment method for pancreatic cancer can be provided.
A combination of the above (A-1) and lapatinib (B-2) is also preferable, and effective in the treatment of breast cancer and non-small cell lung cancer. That is, it is possible to provide a method for treating breast cancer or non-small cell lung cancer including administering each effective dose of (A-1) and lapatinib (B-2). Furthermore, a method for treating breast cancer can be provided by administering capecitabine (C-2) to (A-1) and lapatinib (B-2) and administering each effective dose. Alternatively, a method for treating breast cancer can be provided by administering trastuzumab (C-3) in combination with (A-1) and lapatinib (B-2) and administering each effective dose.
A combination of the above (A-1) and gefitinib (B-3) is also preferable and effective in the treatment of non-small cell lung cancer. That is, the therapeutic method of non-small cell lung cancer including administering each effective dose of this (A-1) and gefitinib (B-3) can be provided.

The present application is based on the discovery that the triazole compound (A) exhibits an action of enhancing the antitumor effect of the EGFR tyrosine kinase inhibitor in cancer treatment using an EGFR tyrosine kinase inhibitor. ) Discloses a novel invention for use as an antitumor effect enhancer of an EGFR tyrosine kinase inhibitor.
In still another aspect of the present application, a method for enhancing the antitumor effect of an EGFR tyrosine kinase inhibitor by administering the triazole compound (A) in combination with an EGFR tyrosine kinase inhibitor is disclosed.
The EGFR tyrosine kinase inhibitor whose antitumor effect is enhanced by the triazole compound (A) has the same meaning as described above. Preferred are erlotinib, lapatinib, gefitinib, afatinib, and neratinib. Particularly preferred are erlotinib, lapatinib and gefitinib.

In order to use the triazole compound (A) for the purpose of enhancing the antitumor effect of the EGFR tyrosine kinase inhibitor, the optimal dose of the EGFR tyrosine kinase inhibitor should be administered daily or intermittently. The triazole compound (A) can be achieved by single, intermittent, or continuous administration of the optimal dose of the triazole compound (A).
It is preferable to use the triazole compound (A) at a dose of 0.1 to 1000 mg per adult. It is preferable to administer this once or intermittently.
The dose per administration of the EGFR tyrosine kinase inhibitor is preferably 1 to 5000 mg based on known pharmacological activity results, and these can be used in appropriate combination. In particular, when erlotinib is used as the EGFR inhibitor, the dose per administration is 50 to 300 mg, which is preferably administered every day. When lapatinib is used, the dose per administration is 500 to 3000 mg, and it is preferably administered every day. When gefitinib is used, the dose per administration is 100 to 500 mg, and it is preferably administered every day. In the administration of the EGFR tyrosine kinase inhibitor, the triazole compound (A) is preferably used at a dose of 0.1 to 1000 mg per dose. The dose of the triazole compound (A) may be appropriately increased or decreased after confirming the reactivity of the antitumor effect.

  That is, according to the present invention, in the treatment of malignant tumors with an EGFR tyrosine kinase inhibitor, by using the triazole compound (A) in combination, the antitumor effect is enhanced and a higher therapeutic effect can be achieved. Alternatively, even for a malignant tumor whose drug sensitivity to an EGFR tyrosine kinase inhibitor is reduced, the drug sensitivity can be recovered by combining the triazole compound (A) with the drug treatment of the EGFR tyrosine kinase inhibitor. Drug treatment with tyrosine kinase inhibitors can be continued for a long time. Furthermore, in the case of obtaining an antitumor effect equivalent to the use of an antitumor agent alone, the dose of each antitumor agent can be relatively reduced, and the occurrence of undesirable pharmacological effects is adversely affected by the antitumor effect. It can be reduced without affecting.

The usefulness of using the triazole compound (A) represented by the general formula (1) of the present invention in combination with the EGFR tyrosine kinase inhibitor (B) will be described in Examples.
1. Reagent Agent As the triazole compound (A) represented by the general formula (1) of the present invention, 5- (2,4-dihydroxy-5-isopropyl-phenyl) -4- [4- (4-methyl-piperazine- 1-ylmethyl) -phenyl] -2,4-dihydro- [1,2,4] triazol-3-one; (A-1) was used. The (A-1) was synthesized according to the production method disclosed in Example 2-5 of International Publication WO2006 / 095783. Further, as a comparative example, 5- (2,4-dihydroxy-5-isopropyl-phenyl) -4- (4-morpholin-4-ylmethyl-phenyl) -isoxazole-3-carboxylic acid, which is an existing HSP90 inhibitor compound Ethylamide (Compound D: NVP-AUY-922) was used. The comparative HSP90 inhibitor (compound D: NVP-AUY-922) is described in J. Am. Med. Chem. , 51 (2), 196-218 (2008), and methanesulfonate was prepared by a conventional method and used for the test. Erlotinib (B-1) was employed as the EGFR tyrosine kinase inhibitor (B). As erlotinib (B-1), Erlotinib Hydrochloride (manufactured by LC Laboratories) was used as it was. As gemcitabine (C-1), 200 mg for injection of Gemzar (registered trademark) was used as it was.
Erlotinib (B-1) was dissolved in 6% Captisol (registered trademark, manufactured by Cydex) and used for administration. Gemcitabine (C-1) was dissolved in Japanese pharmacopoeia physiological saline. Triazole (A-1), which is an HSP90 inhibitor, and a comparative HSP90 inhibitor (Compound D: NVP-AUY-922) were dissolved in a Japanese Pharmacopoeia glucose injection and used for administration.

2. Transplanted tumor A human pancreatic cancer cell line, BxPc-3 (ATCC catalog number, CRL-1687) was grown in RPMI-1640 medium supplemented with 10% FBS. About 1 × 10 ⁷ BxPc-3 cells grown were suspended in HBSS and injected subcutaneously into nude mice. After the cells formed a tumor, the tumor was taken out to produce a tumor piece and transplanted subcutaneously into nude mice using a trocar. The transplantation was repeated by the same operation to maintain the tumor, and a transplanted tumor was prepared.

3. Anti-tumor test using subcutaneous tumor transplanted mice BxPc-3, a human pancreatic cancer tumor maintained by passage in nude mice, was transplanted into the dorsal skin of each nude mouse using a trocar. Reagent administration was started when the tumor volume reached approximately 100-150 mm ³ . The tumor was measured twice a week from the start date of administration to the end of the observation period, and the major axis (L) and minor axis (W) of the tumor were measured using calipers. The tumor volume was calculated from the formula (L × W ² × 1/2) using the measured major axis and minor axis of the tumor.

[Test Example 1]
Using human pancreatic cancer tumor BxPc-3 subcutaneously transplanted nude mice as a human pancreatic cancer xenograft model, the combination therapy of the triazole compound (A-1) and erlotinib (B-1) according to the present invention, and the triazole compound ( A-1), erlotinib (B-1) and gemcitabine (C-1) combined with anti-tumor effect by triazole compound (A-1), erlotinib (B-1), gemcitabine (C-1) and comparison The anti-tumor effects of each administration of the HSP90 inhibitor (Compound D: NVP-AUY-922) and the combined administration of the comparative HSP90 inhibitor (D) and erlotinib (B-1) were compared.
As for the dosage and usage of each drug, triazole (A-1) was administered into the tail vein three times once a week at a dose of 20 mg / kg, which is half the maximum tolerated dose. Erlotinib (B-1) was orally administered once daily for 21 days at a dose of 50 mg / kg. Gemcitabine (C-1) was administered into the tail vein three times once a week at a dose of 150 mg / kg. The comparative HSP90 inhibitor (D) was administered into the tail vein three times once a week at a dose of 40 mg / kg, which is half the maximum tolerated dose. Concomitant administration is a combination administration of triazole (A-1) and erlotinib (B-1) according to the present invention. The triazole compound (A-1) is administered at a dose of 20 mg / kg three times a week, tail vein. And erlotinib (B-1) was orally administered once daily for 21 days at a dose of 50 mg / kg. In addition, the combination administration of triazole (A-1), erlotinib (B-1) and gemcitabine (C-1) according to the present invention is carried out once a week at a dose of 20 mg / kg with the triazole compound (A-1). 3 times in the tail vein, erlotinib (B-1) at a dose of 50 mg / kg orally once daily for 21 days, and gemcitabine (C-1) at 150 mg / kg The dose was administered into the tail vein once a week three times. On the other hand, the combined administration of the comparative HSP90 inhibitor (D) and erlotinib (B-1) was carried out by administering the comparative HSP90 inhibitor (D) into the tail vein three times once a week at a dose of 40 mg / kg, Erlotinib (B-1) was orally administered once daily for 21 days at a dose of 50 mg / kg.
The non-administration group (control group) was added to each drug administration group (group 7), and an anti-tumor test was performed on each group of 5 xenograft models. Table 1 shows the tumor volume of each group on the 21st day from the start of administration, and the relative tumor volume (T / C (%)) with the relative tumor volume of the non-administration group as 100. T / C (%) was calculated by the formula (administration group relative tumor volume / non-administration group relative tumor volume × 100). Further, FIG. 1 shows a tumor growth curve over time in which the relative tumor volume was calculated with the tumor volume on the administration start date being 1.

[Table 1] Anti-tumor effect test results of nude mice transplanted with human pancreatic cancer tumor BxPc-3 subcutaneously

As is apparent from Table 1 and FIG. 1, on the 21st day after the start of administration, erlotinib (B-1) caused a 34% tumor growth suppression relative to the non-administration group. Single administration of the triazole compound (A-1) resulted in 51% growth inhibition. On the other hand, when the triazole compound (A-1) and erlotinib (B-1) were administered in combination, an antitumor effect in which the tumor growth inhibitory action was strongly enhanced to 80% was brought about. On the other hand, the single administration of the comparative HSP90 inhibitor (D) showed only 3% growth suppression at 1/2 the same maximum tolerated dose as the (A-1), and the comparative HSP90 inhibitor (D) and The combined administration of erlotinib (B-1) showed a 46% growth inhibitory effect. Therefore, the combined use of the comparative HSP90 inhibitor (D) and erlotinib (B-1) is a result that the growth inhibition rate of erlotinib (B-1) single administration is only a slight addition effect from 34%. The results were inferior to the antitumor effect of the combined administration of the triazole compound (A-1) and erlotinib (B-1). Although the triazole compound (A-1) is a test at half the maximum tolerated dose, the combined administration of the triazole compound (A-1) and erlotinib (B-1) according to the present invention is remarkable. Excellent anti-tumor effect.
From the above results, the drug treatment administered in combination of the triazole compound (A) and the EGFR tyrosine kinase inhibitor (B) according to the present invention is superior to the combination of the existing HSP90 inhibitor and the EGFR tyrosine kinase inhibitor. It became clear that a tumor effect was obtained.
In this study, administration of gemcitabine (C-1) alone resulted in 56% growth inhibition. In contrast, when the triazole compound (A-1), erlotinib (B-1), and gemcitabine (C-1) are administered in combination, the tumor growth inhibitory action is 96%, and the antitumor effect is significantly enhanced. It was. From the above results, it is shown that the drug treatment administered in combination of the triazole compound (A) according to the present invention, the EGFR tyrosine kinase inhibitor (B), and gemcitabine (C-1) can achieve an excellent antitumor effect. It was.

Claims (11)

General formula (1)

[Wherein, X represents a linear or branched alkyl group having 1 to 8 carbon atoms, an alkynyl group having 2 to 10 carbon atoms, or a halogen atom, Y represents a sulfur atom or an oxygen atom, and m represents 0 An antitumor agent comprising a combination of a triazole compound (A) represented by the following formula: and an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (B) . In the said General formula (1), X is an ethyl group, isopropyl group, tert-butyl group, 2,2-dimethylpropyl group, 2-propynyl group, 2-butynyl group, and a halogen atom. Tumor agent.   In the general formula (1), m is 0 or 1, and A is a morpholino group, a 4-methylpiperazin-1-yl group, a piperidin-1-yl group, or a pyrrolidin-1-yl group. The antitumor agent described. The antitumor agent according to claim 1, wherein the epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (B) is one or more selected from erlotinib, lapatinib, and gefitinib.   Furthermore, the antitumor agent of Claim 1 containing gemcitabine. General formula (1)

[Wherein, X represents a linear or branched alkyl group having 1 to 8 carbon atoms, an alkynyl group having 2 to 10 carbon atoms, or a halogen atom, Y represents a sulfur atom or an oxygen atom, and m represents 0 Inhibition of epidermal growth factor receptor (EGFR) tyrosine kinase, comprising as an active ingredient a triazole compound (A) represented by an integer of ˜4, A represents an amino group having a substituent]] An antitumor effect potentiator that enhances the antitumor effect of the agent. In the said General formula (1), X is an ethyl group, isopropyl group, tert-butyl group, 2,2-dimethylpropyl group, 2-propynyl group, 2-butynyl group, and a halogen atom. Tumor effect enhancer.   In the general formula (1), m is 0 or 1, and A is a morpholino group, a 4-methylpiperazin-1-yl group, a piperidin-1-yl group, or a pyrrolidin-1-yl group. The antitumor effect potentiator described. In the administration of an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor in the treatment of cancer, the general formula (1)

[Wherein, X represents a linear or branched alkyl group having 1 to 8 carbon atoms, an alkynyl group having 2 to 10 carbon atoms, or a halogen atom, Y represents a sulfur atom or an oxygen atom, and m represents 0 An epithelial growth factor receptor (EGFR) tyrosine kinase inhibitor, wherein the triazole compound (A) represented by the following formula is administered in combination: Of enhancing the antitumor effect of. In the said General formula (1), X is an ethyl group, an isopropyl group, a tert- butyl group, a 2, 2- dimethylpropyl group, 2-propynyl group, 2-butynyl group, and a halogen atom. A method for enhancing the antitumor effect of an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor.   In the general formula (1), m is 0 or 1, and A is a morpholino group, a 4-methylpiperazin-1-yl group, a piperidin-1-yl group, or a pyrrolidin-1-yl group. A method for enhancing the antitumor effect of the epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor described in 1. above.

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