JP6580030B2 - Polymer compound in which camptothecin derivative and HSP90 inhibitor are combined and use thereof - Google Patents

Description

  The present invention relates to a polymer compound in which a camptothecin derivative and an HSP90 inhibitor are bound to the same molecule. The polymer compound of the present invention relates to a drug delivery system for simultaneously delivering a camptothecin derivative and an HSP90 inhibitor to an affected area, and also relates to a pharmaceutical compound exhibiting excellent antitumor activity.

Camptothecin is a plant alkaloid extracted from the Chinese plant "Yuki" and is a type I topoisomerase inhibitor. This selectively binds to type I topoisomerase complexed with DNA and stabilizes its structure. As a result, the cleaved DNA cannot be recombined, and is a drug that induces cell death by stopping DNA synthesis.
Camptothecin showed a high antitumor effect and was developed as an anticancer drug in the 1960s. However, clinical trials were canceled due to myelosuppression and hemorrhagic cystitis as strong toxicity.

  Later, topotecan and irinotecan were developed as derivatives that are more soluble in water than camptothecin and have stronger antitumor activity but lower toxicity. Topotecan exerts an antitumor effect without undergoing metabolism, and diarrhea with side effects is mild because 20-40% of the dose is renal excretion. Although irinotecan itself has an antitumor effect, it is metabolized in vivo by carboxylesterase to the active metabolite 7-ethyl-10-hydroxycamptothecin (hereinafter sometimes referred to as EHC) and is stronger. Demonstrate antitumor effect. In addition, irinotecan and EHC are characterized by being present in plasma as a lactone type that is more biologically active than topotecan and having a longer half-life.

  In clinical practice, camptothecin derivatives are used in many cancer types. Topotecan has been approved for use in small cell lung cancer and ovarian cancer that has been pre-treated with cancer chemotherapeutic agents. On the other hand, irinotecan is small cell lung cancer, non-small cell lung cancer, cervical cancer, ovarian cancer, gastric cancer (inoperable or recurrent), colorectal cancer (inoperable or recurrent), breast cancer (inoperable or recurrent), spiny cells. Approved with broad indications for cancer and malignant lymphoma (non-Hodgkin lymphoma).

  In order to efficiently extract the pharmacological activity of an antitumor agent, a technique using a polymer carrier is known, and a polymer-camptothecin derivative conjugate using a camptothecin derivative has been reported. Patent Document 1 describes a polymerized camptothecin derivative prepared by esterifying a camptothecin derivative having a phenolic hydroxyl group with a copolymer in which polyethylene glycols and a polymer having a carboxy group in the side chain are linked. . In this polymerized derivative, it is described that a camptothecin derivative is bound by a phenyl ester bond that is easily chemically cleaved, and a camptothecin derivative having pharmacological activity is slowly released after administration in vivo. . Further, the polymerized camptothecin derivative has a self-association property and forms a micelle-like aggregate to exhibit a high distribution in the tumor tissue, thereby selectively exerting a medicinal effect on the tumor tissue and causing side effects. It is described that there are few antitumor agents. This high molecular weight camptothecin derivative is considered to have an advantage that the release of EHC, which is a medicinal component, does not depend on the enzyme, and thus is hardly affected by individual differences in the therapeutic effect.

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.
HSP is known to interact with various proteins involved in intracellular signal transduction systems. Molecules to which HSP binds are called client proteins, and about 200 molecules of HSP90 client proteins have been reported. That is, HSP90 is often necessary for the functional expression of various proteins, and the mechanism of action is that HSP90 specifically recognizes a client protein in an unstable folded state and binds to it to form a complex. Based on biochemical properties that form. Various proteins (steroid receptors, Raf serine kinases, tyrosine kinases) involved in cancer-related signal transduction depend on the structure of HSP90, and HSP90 regulates the cell cycle, cell canceration / proliferation / survival It is clear that it is deeply involved in the signal.
Human tumors are abnormal in the regulation of many signal molecules, and HSP90 is required to maintain the function of these signal molecules. In recent years, it has been reported that a compound that acts on HSP90 and inhibits its function causes degradation of various client proteins involved in cancer-related signal transduction and can suppress the growth of cancer cells. That is, the HSP90 inhibitor has an action of changing the configuration of a chaperone complex containing a client protein and HSP90. Thereafter, the client protein released from the complex is decomposed mainly in the ubiquitin / proteasome system. As a result, the amount of HSP90 client protein decreases, and accordingly, downstream signal transduction is blocked and cancer cell growth is suppressed, thereby providing an antitumor effect. 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.

Clinical applications of HSP90 inhibitors have been performed previously with geldanamycin derivatives. The geldanamycin derivative 17-AAG has been clinically tested for hematopoietic tumors, breast cancer, melanoma, and gastrointestinal stromal tumor (GIST). There was a problem of developing. Accordingly, the search for a novel low-molecular HSP90 inhibitor and a structure optimization study that can overcome the problems of the geldanamycin derivative were advanced, and as a result, several heterocyclic HSP90 inhibitors were found.
For example, Patent Documents 2 to 4 report 1,2,4-triazol-3-one substituted resorcin derivatives useful as HSP90 inhibitors. Patent Document 5 discloses polymerized 1,2 which is a polymerized HSP90 inhibitor aimed at improving the pharmacokinetics of 1,2,4-triazol-3-one-substituted resorcin derivatives and the sustained release of drugs. 1,4-triazol-3-one substituted resorcin derivatives have been described. In the polymerized HSP90 inhibitor, a 1,2,4-triazol-3-one substituted resorcin derivative, which is an HSP90 inhibitor, was bound to a block copolymer having a polyethylene glycol segment and a polymer segment having a carboxy group. It is a polymerized drug-binding compound. This is an antitumor agent that has self-association properties, shows micelle-like aggregates, and shows high distribution in the tumor tissue, so that the drug is selectively acted on the tumor tissue and has a medicinal effect, and has few side effects It is described that. That is, when the polymerized HSP90 inhibitor is administered to a living body, the 1,2,4-triazol-3-one substituted resorcin derivative having a high concentration in the tumor tissue and having HSP90 inhibitory activity is gradually released to produce an antitumor agent. It is described as an antitumor agent that exerts an effect.

The anti-tumor effect is expected to be enhanced by combining the HSP90 inhibitor with a cell-killing antitumor agent, and the development of cancer treatment using a combination of the HSP90 inhibitor and another anti-tumor agent has been promoted. Yes.
Patent Document 6 describes the combined use of a 1,2,4-triazol-3-one-substituted resorcin derivative that is an HSP90 inhibitor and camptothecin that is a type I topoisomerase inhibitor. Non-Patent Document 1 reports a phase I clinical trial using a combination of 17-AAG, which is an HSP90 inhibitor, and irinotecan.

  Co-administration of a plurality of drugs having different pharmacological activity mechanisms may have a synergistic pharmacological activity enhancing effect, and various combinations are being studied in order to obtain a high therapeutic effect. However, in order for the synergistic pharmacological activity enhancing effect at the cellular level to be effective against an actual disease, it is necessary to cause a plurality of drugs to act on the diseased affected area at a desired concentration and sensitization time. There is a need to develop drug delivery systems for delivering drugs. Patent Document 7 and Non-Patent Document 2 report a polymer compound retaining doxorubicin and another anticancer agent. However, there are no reports on drug delivery compounds that take into account optimal drug delivery of two drugs in combination therapy of camptothecin derivatives and HSP90 inhibitors.

International Publication WO2004 / 039869 Pamphlet International Publication WO2006 / 055760 Pamphlet International Publication WO2006 / 087077 Pamphlet International Publication WO2006 / 095783 Pamphlet Dating published in WO2008 / 041610 pamphlet International Publication WO2013 / 067162 Pamphlet Special table 2010-519305 gazette

Tse A. N. et al. , Clin. Cancer Res. 14:20, 6704-6711 (2008) Younso Bae & Kazunori Kataoka, Adv. Drug Deliv. Rev. 61,768-784 (2009)

  It is an object of the present invention to efficiently deliver a camptothecin derivative and an HSP90 inhibitor, particularly a 1,2,4-triazol-3-one substituted resorcin derivative, simultaneously to a diseased site to release a drug, and to achieve a desired concentration and sensitization. By providing a drug delivery system (DDS) that acts over time, it is an object of the present invention to provide a pharmaceutical compound that suppresses cell damage to normal cells, reduces side effects, and provides an excellent antitumor effect.

［式中、Ｒ_１は水素原子又は置換基を有していてもよい（Ｃ１〜Ｃ４）アルキル基を示し、ｔは４５〜４５０の整数を示し、Ａは（Ｃ１〜Ｃ６）アルキレン基であり、ｄ＋ｅ＋ｆ＋ｇ＋ｈは６〜６０の整数を示し、ｄ＋ｅ＋ｆ＋ｇ＋ｈに対するｄの割合が５〜５０％、ｅの割合が５〜９０％、ｆの割合が０〜９０％、ｇの割合が０〜９０％、ｈの割合が０〜９０％であり、Ｒ_２は水素原子または（Ｃ１〜Ｃ４）アシル基を示し、Ｒ_３はＨＳＰ９０阻害剤の結合残基またはＨＳＰ９０阻害剤が結合したアスパラギン酸残基を示し、Ｒ_４はアスパラギン酸残基またはアスパラギン酸イミド残基を示し、Ｒ_５は−Ｎ（Ｒ_６）ＣＯＮＨ（Ｒ_７）を示し、Ｒ_６及びＲ_７は同一でも異なってもいてもよく、分岐若しくは環状の（Ｃ３〜Ｃ６）アルキル基または三級アミノ基で置換されていてもよい分岐若しくは直鎖の（Ｃ１〜Ｃ５）アルキル基を示し、前記ポリグルタミン酸セグメントは、側鎖カルボキシ基に７−エチル−１０−ヒドロキシカンプトテシンが結合したグルタミン酸単位、側鎖カルボキシ基にＲ_３基が結合したグルタミン酸単位、側鎖カルボキシ基にＲ_４基が結合したグルタミン酸単位、側鎖カルボキシ基にＲ_５基が結合したグルタミン酸単位及び側鎖カルボキシ基が遊離カルボキシ基であるグルタミン酸単位が、それぞれ独立して、ランダムに配列したポリグルタミン酸構造である。］で表される高分子化合物。The present inventors have created a polymer compound in which a camptothecin derivative and an HSP90 inhibitor, particularly a 1,2,4-triazol-3-one substituted resorcin derivative having HSP90 inhibitory activity, are bonded to the same molecule, The present inventors have found that a compound can provide a satisfactory therapeutic effect as a chemotherapeutic agent, and have reached the present invention.
That is, the gist of the present invention is the following configurations 1) to 8).
1) A polymer compound in which a camptothecin derivative and an HSP90 inhibitor are bound to a block copolymer in which a polyethylene glycol segment and a polyglutamic acid segment are linked, and the polymer compound has the general formula (1)

[Wherein R ₁ represents a hydrogen atom or an optionally substituted (C1-C4) alkyl group, t represents an integer of 45-450, and A represents a (C1-C6) alkylene group. , D + e + f + g + h represents an integer of 6-60, the ratio of d to d + e + f + g + h is 5-50%, the ratio of e is 5-90%, the ratio of f is 0-90%, the ratio of g is 0-90%, h the ratio is 0 to 90%, _{R 2} represents a hydrogen atom or a (C1 -C4) acyl group, _{R 3} represents aspartic acid residue bound binding residue or HSP90 inhibitors of HSP90 inhibitors, R ₄ represents an aspartic acid residue or an aspartic imide residue, R ₅ represents —N (R ₆ ) CONH (R ₇ ), and R ₆ and R ₇ may be the same or different, branched or Cyclic (C3-C6) Al A branched or straight-chain (C1-C5) alkyl group which may be substituted with a ruthenium group or a tertiary amino group, and the polyglutamic acid segment has 7-ethyl-10-hydroxycamptothecin bonded to a side chain carboxy group Glutamic acid unit, glutamic acid unit in which R ₃ group is bonded to the side chain carboxy group, glutamic acid unit in which R ₄ group is bonded to the side chain carboxy group, glutamic acid unit in which R ₅ group is bonded to the side chain carboxy group, and side chain carboxy group Is a polyglutamic acid structure in which the glutamic acid units, each of which is a free carboxy group, are independently and randomly arranged. ] The high molecular compound represented.

［式中、Ｘ_１は直鎖若しくは分岐の（Ｃ１〜Ｃ６）のアルキル基または−ＣＯＮＲ_１１Ｒ_１２を示し、前記Ｒ_１１及びＲ_１２は同一でも異なっていてもよい（Ｃ１〜Ｃ６）の直鎖状、分岐状または環状アルキル基であり、Ｘ_２は置換基を有していてもよいアリール基を示す。］で示される１，２，４−トリアゾール−３−オン置換レゾルシン誘導体であり、レゾルシン部分の水酸基がエステル結合したＨＳＰ９０阻害剤の結合残基である、前記１）に記載の高分子化合物。2) R ₃ is a binding residue of an HSP90 inhibitor, and the HSP90 inhibitor is represented by the general formula (2)

[Wherein, X ₁ represents a linear or branched (C 1 to C 6) alkyl group or —CONR ₁₁ R ₁₂ , and R ₁₁ and R ₁₂ may be the same or different (C 1 to C 6) It is a chain, branched or cyclic alkyl group, and X ₂ represents an aryl group which may have a substituent. The polymer compound according to 1) above, which is a 1,2,4-triazol-3-one-substituted resorcin derivative represented by formula (I), wherein the hydroxyl group of the resorcin moiety is a binding residue of an HSP90 inhibitor having an ester bond.

［式中、Ｒ_８及びＲ_９はそれぞれ独立して水素原子または（Ｃ１〜Ｃ６）アルキル基を示し、Ｒ_１０は水素原子、（Ｃ１〜Ｃ４０）アルキル基、（Ｃ１〜Ｃ４０）アラルキル基、芳香族基及びカルボキシ基が保護されたアミノ酸残基からなる群から選択される１種以上の置換基を示し、ＣＸ−ＣＹはＣＨ−ＣＨ若しくはＣ＝Ｃ（二重結合）を示し、ＤはＨＳＰ９０阻害剤の結合残基を示す。］である、前記１）に記載の高分子化合物。3) R ₃ is an aspartic acid residue to which an HSP90 inhibitor is bound, and the aspartic acid residue is represented by the general formula (3) or (4)

[Wherein R ₈ and R ₉ each independently represent a hydrogen atom or a (C1-C6) alkyl group, and R ₁₀ represents a hydrogen atom, a (C1-C40) alkyl group, a (C1-C40) aralkyl group, an aromatic group. One or more substituents selected from the group consisting of amino acid residues in which a group group and a carboxy group are protected, CX-CY represents CH-CH or C = C (double bond), and D represents HSP90. The binding residues of the inhibitor are indicated. ] The polymer compound according to 1) above.

［式中、Ｘ_１は直鎖若しくは分岐の（Ｃ１〜Ｃ６）のアルキル基または−ＣＯＮＲ_１１Ｒ_１２を示し、前記Ｒ_１１及びＲ_１２は同一でも異なっていてもよい直鎖状、分岐状または環状の（Ｃ１〜Ｃ６）アルキル基であり、Ｘ_２は置換基を有していてもよいアリール基を示す。］で示される１，２，４−トリアゾール−３−オン置換レゾルシン誘導体であり、レゾルシン部分の水酸基がエステル結合したＨＳＰ９０阻害剤の結合残基である、前記３）に記載の高分子化合物。4) The HSP90 inhibitor of the binding residue of the HSP90 inhibitor of D is represented by the general formula (2)

[Wherein, X ₁ represents a linear or branched (C1-C6) alkyl group or —CONR ₁₁ R ₁₂ , wherein R ₁₁ and R ₁₂ may be the same or different, and may be linear, branched or an annular (C1 -C6) alkyl group, X ₂ represents an aryl group which may have a substituent. The polymer compound according to 3) above, which is a 1,2,4-triazol-3-one-substituted resorcin derivative represented by formula (I), wherein the hydroxyl group of the resorcin moiety is a binding residue of an HSP90 inhibitor having an ester bond.

5) The polymer compound according to any one of 1) to 4), wherein a ratio of d to e in the general formula (1) is d: e = 1: 0.5 to 15.
6) R ₁ is a methyl group or ethyl group, A is an ethylene group or propylene group, R ₂ is an acetyl group or propionyl group, R ₆ and R ₇ are both cyclohexyl groups or both isopropyl groups, and the ratio of d to d + e + f + g + h is 5) to 40%, 5 to 80% of e, 0 to 60% of f, 5 to 40% of g, and 0 to 30% of h, 1) to 5) above The high molecular compound as described in any one.
7) An antitumor agent comprising the polymer compound according to any one of 1) to 6) as an active ingredient.
8) The antitumor agent according to 7) above, which is prescribed for a malignant tumor treated with an anticancer treatment with a camptothecin derivative in the previous treatment.

  In the polymer compound of the present invention, a camptothecin derivative that is a key drug in cancer chemotherapy is bound to an HSP90 inhibitor, particularly a 1,2,4-triazol-3-one substituted resorcin derivative having HSP90 inhibitory activity. It is possible to provide a drug delivery system that can deliver two drugs, that is, a camptothecin derivative and an HSP90 inhibitor, to the same site in an optimal amount and in an optimal time. The drug delivery system of the present invention can deliver two drugs to the affected area at the same time, and can act on the drug at an ideal drug concentration and drug sensitization time, thus producing a synergistic effect of the two drugs. Enables theoretical combination therapy and can achieve enhanced antitumor effects and reduced side effects. Thus, efficient and safe cancer chemotherapy can be achieved.

The ratio of the released amount of EHC and compound 5 to the total amount of bound drug at 37 ° C. in PBS solution of compound 1 (phosphate buffered saline; pH 7.1) is shown. The ratio of the released amount of EHC and Compound 5 to the total amount of bound drug at 37 ° C. in PBS solution of Compound 2 (phosphate buffered saline; pH 7.1) is shown. The ratio of the released amounts of EHC and compound 5 to the total amount of bound drug at 37 ° C. in PBS solution of compound 3 (phosphate buffered saline; pH 7.1) is shown. The change of the relative tumor volume from the administration start date when the high molecular compound (compound 1) of this invention and the control drugs 1 and 2 are administered to the mouse | mouth colon cancer Colon38 transplantation mouse | mouth is shown.

［式中、Ｒ_１は水素原子又は置換基を有していてもよい（Ｃ１〜Ｃ４）アルキル基を示し、ｔは４５〜４５０の整数を示し、Ａは（Ｃ１〜Ｃ６）アルキレン基であり、ｄ＋ｅ＋ｆ＋ｇ＋ｈは６〜６０の整数を示し、ｄ＋ｅ＋ｆ＋ｇ＋ｈに対するｄの割合が５〜５０％、ｅの割合が５〜９０％、ｆの割合が０〜９０％、ｇの割合が０〜９０％、ｈの割合が０〜９０％であり、Ｒ_２は水素原子または（Ｃ１〜Ｃ４）アシル基を示し、Ｒ_３はＨＳＰ９０阻害剤の結合残基またはＨＳＰ９０阻害剤が結合したアスパラギン酸残基を示し、Ｒ_４はアスパラギン酸残基またはアスパラギン酸イミド残基を示し、Ｒ_５は−Ｎ（Ｒ_６）ＣＯＮＨ（Ｒ_７）を示し、Ｒ_６、Ｒ_７は同一でも異なってもいてもよく、（Ｃ３〜Ｃ６）分岐若しくは環状アルキル基または三級アミノ基で置換されていてもよい（Ｃ１〜Ｃ５）分岐若しくは直鎖アルキル基を示し、前記ポリグルタミン酸セグメントは、側鎖カルボキシ基に７−エチル−１０−ヒドロキシカンプトテシンが結合したグルタミン酸単位、側鎖カルボキシ基にＲ_３基が結合したグルタミン酸単位、側鎖カルボキシ基にＲ_４基が結合したグルタミン酸単位、側鎖カルボキシ基にＲ_５基が結合したグルタミン酸単位及び側鎖カルボキシ基が遊離カルボキシル基であるグルタミン酸単位がそれぞれ独立して、ランダムに配列したポリグルタミン酸構造である。］で表される。The present invention is a block copolymer in which a polyethylene glycol segment and a polyglutamic acid segment in which a camptothecin derivative and an HSP90 inhibitor are bonded to the same molecule are linked, and the general formula (1)

[Wherein R ₁ represents a hydrogen atom or an optionally substituted (C1-C4) alkyl group, t represents an integer of 45-450, and A represents a (C1-C6) alkylene group. , D + e + f + g + h represents an integer of 6-60, the ratio of d to d + e + f + g + h is 5-50%, the ratio of e is 5-90%, the ratio of f is 0-90%, the ratio of g is 0-90%, h the ratio is 0 to 90%, _{R 2} represents a hydrogen atom or a (C1 -C4) acyl group, _{R 3} represents aspartic acid residue bound binding residue or HSP90 inhibitors of HSP90 inhibitors, R ₄ represents an aspartic acid residue or an aspartic imide residue, R ₅ represents —N (R ₆ ) CONH (R ₇ ), and R ₆ and R ₇ may be the same or different, (C3 To C6) branched or cyclic alkyl Or a (C1-C5) branched or straight chain alkyl group optionally substituted with a tertiary amino group, wherein the polyglutamic acid segment is a glutamic acid unit in which 7-ethyl-10-hydroxycamptothecin is bound to a side chain carboxy group A glutamic acid unit having an R ₃ group bonded to a side chain carboxy group; a glutamic acid unit having an R ₄ group bonded to a side chain carboxy group; a glutamic acid unit having an R ₅ group bonded to a side chain carboxy group; It is a polyglutamic acid structure in which glutamic acid units as groups are independently arranged at random. ].

In the general formula (1), R ₁ is a terminal modification group of the polyethylene glycol segment, and is a hydrogen atom or a (C1-C4) alkyl group which may have a substituent.
The (C1-C4) alkyl group which may have a substituent in R ₁ is a linear or branched (C1-C4) alkyl group. Specific examples are methyl group, ethyl group, propyl group, isopropyl group, n-butyl group, isobutyl group, sec-butyl group or tert-butyl group. Examples of the substituent of the (C1 to C4) alkyl group which may have a substituent include an amino group, a dialkylamino group, an alkyloxy group, and a carboxyl group. R ₁ is preferably a methyl group or an ethyl group, and particularly preferably a methyl group.

In the polyethylene glycol segment in the general formula (1), t in the general formula (1) indicating the number of unit repeating structures of an ethyleneoxy group; (OCH ₂ CH ₂ ) group is 45 to 450, preferably 90 to 340. is there. That is, the molecular weight of the polyethylene glycol moiety is preferably 2,000 to 20,000, more preferably 4,000 to 15,000.

  A in the general formula (1) is a linking group that connects the polyethylene glycol segment and the polyglutamic acid segment. A in the general formula (1) is an alkylene group of (C1 to C6), and examples thereof include a methylene group, an ethylene group, a trimethylene group, a tetramethylene group, and a hexamethylene group. Among them, an ethylene group or a trimethylene group is preferable, and a trimethylene group is particularly preferable.

  The polyglutamic acid segment of the polymer compound of the present invention represented by the general formula (1) is a structure in which glutamic acid units are amide-bonded. The amide bond is mainly a structure bonded with an α-amide bond type, but may include a structure bonded with a γ-amide bond type. The glutamic acid of each glutamic acid unit may be L-type or D-type. The total number of glutamic acid units (number of polyglutamic acid polymerizations) in the general formula (1) is represented by d + e + f + g + h and is 6 to 60. Preferably d + e + f + g + h is 8-40. Therefore, the average molecular weight of the polyglutamic acid segment is about 600 to 15,000, preferably about 800 to 10,000.

As R < ₂ > in General formula (1), a hydrogen atom or a (C1-C4) acyl group is mentioned. R ₂ is preferably a (C1 to C4) acyl group, and examples thereof include a formyl group, an acetyl group, and a propionyl group. Among them, an acetyl group or a propionyl group is preferable, and an acetyl group is particularly preferable.

R ₅ in the general formula (1) is a urea structure substituent represented by —N (R ₆ ) CONH (R ₇ ), and R ₆ and R ₇ may be the same or different, and may be branched or cyclic (C 3 -C6) A branched or linear (C1-C5) alkyl group optionally substituted with an alkyl group or a tertiary amino group. Examples of the branched or cyclic (C3 to C6) alkyl group include isopropyl group, isobutyl group, sec-butyl group, tert-butyl group, 1-methylbutyl group, 2-methylbutyl group, neopentyl group, cyclohexyl group and the like. More preferred are isopropyl group and cyclohexyl group. Examples of the branched or straight chain (C1-C5) alkyl group which may be substituted with the tertiary amino group include an ethyl group and a dimethylaminopropyl group. As R ₆ and R _7, it is preferable that R ₆ and R ₇ are both an isopropyl group or a cyclohexyl group, or R ₆ and R ₇ are a combination of an ethyl group and a dimethylaminopropyl group. A substituent in which R ₆ and R ₇ are both an isopropyl group or a cyclohexyl group is particularly preferable.

  As shown in the general formula (1), 7-ethyl-10-hydroxycamptothecin (EHC), which is a camptothecin derivative, is included in the polyglutamic acid segment of the polymer compound of the present invention. The chain carboxy group is ester linked.

［式中、Ｘ_１は直鎖若しくは分岐の（Ｃ１〜Ｃ６）アルキル基または−ＣＯＮＲ_１１Ｒ_１２を示し、前記Ｒ_１１及びＲ_１２は同一でも異なってもいてもよい直鎖状、分岐状または環状の（Ｃ１〜Ｃ６）アルキル基であり、Ｘ_２は置換基を有していてもよいアリール基を示す。］で示される１，２，４−トリアゾール−３−オン置換レゾルシン誘導体を挙げることができる。R ₃ in the general formula (1) is a binding residue of the HSP90 inhibitor or an aspartic acid residue to which the HSP90 inhibitor is bound. In the present invention, the HSP90 inhibitor is a drug that inhibits the binding of HSP90 to the client protein by binding to the ATP binding site of HSP90, and finally suppresses cell proliferation, such as a geldanamycin derivative, There are several developed compounds such as substituted resorcin derivatives.
Particularly preferred is a substituted resorcin derivative having a hydroxyl group that can be bonded to the polyglutamic acid segment by an ester bond. Examples of the substituted resorcin derivative include those represented by the general formula (2)

[Wherein, X ₁ represents a linear or branched (C ₁ -C 6) alkyl group or —CONR ₁₁ R ₁₂ , and R ₁₁ and R ₁₂ may be the same or different, and may be linear, branched or an annular (C1 -C6) alkyl group, X ₂ represents an aryl group which may have a substituent. And a 1,2,4-triazol-3-one substituted resorcin derivative represented by the formula:

X ₁ in the general formula (2) represents a linear or branched (C1 to C6) alkyl group or —CONR ₁₁ R ₁₂ , and the R ₁₁ and R ₁₂ may be the same or different, It is a branched or cyclic (C1-C6) alkyl group. Examples of the linear or branched (C1-C6) alkyl group in X ₁ include a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, a sec-butyl group, an isobutyl group, and a tert-butyl group. 1-pentyl group, 1-ethyl-propyl group, 1-hexyl group and the like. Examples of the linear, branched or cyclic (C1-C6) alkyl group which may be the same or different in R ₁₁ and R ₁₂ include a methyl group, an ethyl group, an n-propyl group, an isopropyl group, and an n-butyl group. Group, sec-butyl group, isobutyl group, tert-butyl group, 1-pentyl group, 1-ethyl-propyl group, 1-hexyl group, cyclohexyl group and the like. X ₁ is an isopropyl group, sec-butyl group, isobutyl group, tert-butyl group or —CONR ₁₁ R ₁₂ , preferably R ₁₁ is a methyl group and R ₁₂ is an n-butyl group.

The X ₂ group in the general formula (2) is an aryl group which may have a substituent. Examples of the aryl group which may have a substituent include a carbocyclic aryl group which may have a substituent and a heterocyclic aryl group which may have a substituent.

In the carbocyclic aryl group which may have a substituent for X _2, examples of the carbocyclic aryl group include a phenyl group and a naphthyl group. The carbocyclic aryl group which may have a substituent includes a carbocyclic aryl group having an alkyl group as a substituent, a carbocyclic aryl group having a halogen atom as a substituent, and a carbocyclic aryl having an alkoxy group as a substituent. And a carbocyclic aryl group having an amino group as a substituent, or a carbocyclic aryl group having an aminoalkyl group as a substituent.

The carbocyclic aryl group having an alkyl group as the substituent is a carbocyclic aryl group in which at least one linear, branched, or cyclic (C1-C30) alkyl group is substituted. Preferably it is a phenyl group substituted by a linear or branched (C1-C8) alkyl group. For example, 2-methylphenyl group, 4-methylphenyl group, 3-isopropylphenyl group, 4-isopropylphenyl group, n-hexylphenyl group, n-octylphenyl group, 1-ethylpropylphenyl group, 2-methylpropylphenyl Groups and the like.
The carbocyclic aryl group having a halogen atom as the substituent is a carbocyclic aryl group in which at least one halogen atom is substituted. A monohalogen-substituted phenyl group or a dihalogen-substituted phenyl group is preferable. Examples include 2-chlorophenyl group, 4-chlorophenyl group, 2,4-dichlorophenyl group, 2-bromophenyl group, 4-bromophenyl group, 2,4-dibromophenyl group, and the like.
The carbocyclic aryl group having an alkoxy group as the substituent is a carbocyclic aryl group having at least one primary, secondary, or tertiary (C1-C10) alkoxy group. For example, 4-methoxyphenyl group, 3-methoxyphenyl group, 3,4-dimethoxyphenyl group, 3,4-methylenedioxyphenyl group can be mentioned. Of these, a 4-methoxyphenyl group is preferable.

The carbocyclic aryl group having an amino group as a substituent in X ₂ includes, as the amino group, an unsubstituted amino group, an acyclic primary or secondary amino group, or a cyclic secondary amino group as a substituent. And a carbocyclic aryl group provided.
In the amino group provided as a substituent, examples of the non-cyclic primary amino group include a linear, branched or cyclic (C1-C10) alkyl group, or an amino group substituted with an aryl group. For example, methylamino group, isopropylamino group, neopentylamino group, n-hexylamino group, cyclohexylamino group, n-octylamino group, phenylamino group and the like can be mentioned.
The acyclic secondary amino group may be the same or different, and is a linear, branched or cyclic (C1-C10) alkyl group, or an amino group in which an aryl group is N, N-disubstituted. It is a group. Examples thereof include a dimethylamino group, a diisopropylamino group, an N-methyl-N-cyclohexylamino group, an N-methyl-N-phenylamino group, an N-methyl-N-pyridylamino group, and a diphenylamino group.
Examples of the cyclic secondary amino group include a morpholino group, a piperazin-1-yl group, a 4-methylpiperazin-1-yl group, a piperidin-1-yl group, and a pyrrolidin-1-yl group.

The amino group of the substituent is preferably an acyclic aliphatic primary or acyclic secondary amino group or a cyclic aliphatic secondary amino group. That is, preferred examples of the acyclic aliphatic primary amino group include a methylamino group, isopropylamino group, neopentylamino group, n-hexylamino group, cyclohexylamino group, and n-octylamino group. .
Preferred examples of the acyclic aliphatic secondary amino group include a dimethylamino group, a diisopropylamino group, and an N-methyl-N-cyclohexylamino group.
Preferred examples of the cyclic aliphatic secondary amino group include a morpholino group, a piperazin-1-yl group, a 4-methylpiperazin-1-yl group, a piperidin-1-yl group, and a pyrrolidin-1-yl group. Can do.
Examples of the carbocyclic aryl group in the carbocyclic aryl group having an amino group as a substituent include a phenyl group and a naphthyl group. The carbocyclic aryl group is preferably a phenyl group. The substitution position of the amino group on the phenyl group is not particularly limited, and may be any substitution product at the 2-6 position. A substituted amino group at the 3-position or 4-position is preferred.
In the carbocyclic aryl group having an amino group as a substituent, a preferred embodiment is a phenyl group in which an acyclic aliphatic secondary amino group is substituted at the 4-position, or a cyclic aliphatic secondary amino group at the 4-position. Examples include substituted phenyl groups. In particular, 4-dimethylaminophenyl group, 4- (morpholino) phenyl group, or 4- (4-methylpiperazin-1-yl) phenyl group is preferable.

The carbocyclic aryl group having an aminoalkyl group as a substituent for X ₂ includes, as the amino group, an unsubstituted amino group, an acyclic primary or acyclic secondary amino group, or a cyclic secondary amino group And a carbocyclic aryl group substituted with a (C1-C8) alkyl group having a substituent.
Examples of the non-cyclic primary amino group include a linear, branched or cyclic (C1-C10) alkyl group, or an amino group substituted with an aryl group. For example, methylamino group, isopropylamino group, neopentylamino group, n-hexylamino group, cyclohexylamino group, n-octylamino group, phenylamino group and the like can be mentioned.
The acyclic secondary amino group may be the same or different, and is a linear, branched or cyclic (C1-C10) alkyl group, or an amino group in which an aryl group is N, N-disubstituted. It is a group. Examples thereof include a dimethylamino group, a diisopropylamino group, an N-methyl-N-cyclohexylamino group, an N-methyl-N-phenylamino group, an N-methyl-N-pyridylamino group, and a diphenylamino group.
Examples of the cyclic secondary amino group include a morpholino group, a piperazin-1-yl group, a 4-methylpiperazin-1-yl group, a piperidin-1-yl group, and a pyrrolidin-1-yl group.

The amino group of the substituent is preferably an acyclic aliphatic primary or acyclic secondary amino group or a cyclic aliphatic secondary amino group. That is, preferred examples of the acyclic aliphatic primary amino group include a methylamino group, isopropylamino group, neopentylamino group, n-hexylamino group, cyclohexylamino group, and n-octylamino group. .
Preferred examples of the acyclic aliphatic secondary amino group include a dimethylamino group, a diisopropylamino group, and an N-methyl-N-cyclohexylamino group.
Preferred examples of the cyclic aliphatic secondary amino group include a morpholino group, a piperazin-1-yl group, a 4-methylpiperazin-1-yl group, a piperidin-1-yl group, and a pyrrolidin-1-yl group. Can do.
Examples of the (C1-C8) alkyl group having the amino group as a substituent include a methyl group, an ethyl group, a propyl group, a butyl group, a pentyl group, and an octyl group.
Examples of the carbocyclic aryl group having an aminoalkyl group as a substituent include a phenyl group and a naphthyl group. The carbocyclic aryl group is preferably a phenyl group. The substitution position of the aminoalkyl group on the phenyl group is not particularly limited, and may be any substitution product at the 2-6 position. A substituted amino group at the 3-position or 4-position is preferred.
In the carbocyclic aryl group having an aminoalkyl group as a substituent, as a preferred embodiment, a cyclic aliphatic secondary amino group is substituted at the terminal position of the (C1-C5) alkyl group, and the other terminal group is a phenyl group. In which the 4-position is substituted. In particular, 4- (morpholinomethyl) phenyl group, 4- (4-methylpiperazin-1-ylmethyl) phenyl group, 4- (2-morpholinoethyl) phenyl group, 4- [2- (4-methylpiperazine- A 1-yl) ethyl] phenyl group, a 4- (4-morpholinobutyl) phenyl group, and a 4- [5- (4-methylpiperazin-1-yl) pentyl] phenyl group are preferred. A more preferred embodiment is a 4- (morpholinomethyl) phenyl group or a 4- (4-methylpiperazin-1-ylmethyl) phenyl group.

The heterocyclic aryl group which may have a substituent for X ₂ includes a pyridyl group which may have a substituent, a pyrimidinyl group which may have a substituent, and a substituent. May have a quinolyl group, a quinazolinyl group which may have a substituent, a naphthyridinyl group which may have a substituent, a furyl group which may have a substituent, or a substituent A pyrrolyl group, an indolyl group which may have a substituent, an imidazolyl group which may have a substituent, a pyrazolyl group which may have a substituent, an oxazolyl group which may have a substituent , An isoxazolyl group which may have a substituent, a triazolyl group which may have a substituent, and the like. Preferably, a pyridyl group which may have a substituent, a pyrimidinyl group which may have a substituent, an indolyl group which may have a substituent, an imidazolyl group which may have a substituent Can be mentioned. Particularly preferred are a pyridyl group which may have a substituent, a pyrimidinyl group which may have a substituent, and an indolyl group which may have a substituent.

That is, as a preferable aspect of the heterocyclic aryl group which may have a substituent in X ₂ in the present invention, a pyridyl group, a pyridyl group having an alkyl group as a substituent, and a pyridyl group having an amino group as a substituent A pyrimidinyl group having an alkyl group as a substituent, a pyrimidinyl group having an amino group as a substituent, an indolyl group having an alkyl group as a substituent, and an indolyl group having an amino group as a substituent.
Examples of the pyridyl group having an alkyl group as a substituent include a 6-methylpyridin-3-yl group and a 5-methylpyridin-2-yl group. Examples of the pyridyl group having an amino group as a substituent include a 5-dimethylaminopyridin-2-yl group. Examples of the pyrimidinyl group having an alkyl group as a substituent include 2-methylpyrimidin-4-yl group and 2-methylpyrimidin-3-yl group. Examples of the pyrimidinyl group having an amino group as a substituent include 2-dimethylaminopyrimidin-5-yl group, 2-morpholinopyrimidin-5-yl group, and 2- (4-methylpiperazin-1-yl) pyrimidine-5 Yl group and the like. Examples of the indolyl group having an alkyl group as a substituent include a 1-methylindol-5-yl group and a 1-ethylindol-5-yl group.

In a 1,2,4-triazol-3-one substituted resorcin derivative preferable as an HSP90 inhibitor in the present invention, X ₁ is an isopropyl group or —CONR ₁₁ R ₁₂ , R ₁₁ is a methyl group, and R ₁₂ is n-butyl. It is a group. X ₂ is selected from the group consisting of 2-methylphenyl group, 4- (morpholinomethyl) phenyl group, 4- (4-methylpiperazin-1-ylmethyl) phenyl group, and 1-methylindol-5-yl group. The substituent compounds are preferred.
As an HSP inhibitor that binds to the block copolymer, preferably, 5- (2,4-dihydroxy-5-isopropyl, wherein X ₁ is an isopropyl group and X ₂ is a 1-methylindol-5-yl group. Phenyl) -4- (1-methyl-1H-indol-5-yl) -2,4-dihydro- [1,2,4] triazol-3-one (the following general formula (5); compound 5); X 5- (2,4-dihydroxy-5- (N-methyl), wherein ₁ is -CONR ₁₁ R ₁₂ , R ₁₁ is a methyl group, R ₁₂ is an n-butyl group, and X ₂ is a 2-methylphenyl group -N-butyramide))-4- (2-methylphenyl) -2,4-dihydro- [1,2,4] triazol-3-one (the following general formula (6); compound 6); or X ₁ There an isopropyl group, _{X 2} is 4- 5- (2,4-dihydroxy-5-isopropylphenyl) -4- [4- (4-methylpiperazin-1-ylmethyl) phenyl] -2,4 which is a 4-methylpiperazin-1-ylmethyl) phenyl group -Dihydro- [1,2,4] triazol-3-one dihydrochloride (the following general formula (7); compound 7) and the like can be mentioned.
The compound serving as a substituent for R ₃ may be of a single type or a plurality of types within the same polymer or between molecules of the compound represented by formula (1), but it must be a single type. Is preferred.

In the 1,2,4-triazol-3-one-substituted resorcin derivative represented by the general formula (2), each compound of the combination of the X ₁ group and the X ₂ group is synthesized based on known literature. be able to. The following documents can be cited as known documents for the synthesis of the 1,2,4-triazol-3-one substituted resorcin derivatives.
The compound 5 represented by the general formula (5) is a known compound, and can be produced, for example, according to the description in International Publication No. 2007/139552.
The compound 6 represented by the general formula (6) is a known compound, and can be produced, for example, according to the description in International Publication No. 2008/086857.
Compound 7 represented by the general formula (7) is a known compound, and can be produced, for example, according to the description in International Publication No. 2006/095783.

  The polymer compound in the present invention has a structure in which an HSP90 inhibitor is bound to a polyglutamic acid segment. That is, the HSP90 inhibitor is bound using the carboxylic acid side chain of the polyglutamic acid segment. The binding mode is not particularly limited, and the carboxylic acid side chain of the polyglutamic acid segment and the substituent of the HSP90 inhibitor may be chemically bonded in an appropriate binding mode. Preferably, the polyglutamic acid segment and the HSP90 inhibitor are preferably bonded via an ester bond. For example, when the HSP90 inhibitor has a phenolic hydroxyl group in the molecule, the phenolic hydroxyl group and the side chain carboxy group of the polyglutamic acid segment may be bound by an ester bond. When the polymer compound according to the present invention having an ester bond using a phenolic hydroxyl group is administered in vivo, the ester bond is gradually cleaved non-enzymatically. That is, the release of the HSP90 inhibitor from the polymer compound of the present invention can be controlled to cause sustained release. Since the 1,2,4-triazol-3-one-substituted resorcin derivative preferable as the HSP90 inhibitor has a phenolic hydroxyl group in the resorcin moiety, the resorcin hydroxyl group and the side chain carboxy group of the polyglutamic acid segment are ester-bonded. A mode of bonding by the above can be used.

  As another embodiment, when the HSP90 inhibitor does not have a phenolic hydroxyl group in the molecule, the polyglutamic acid segment is bound to the HSP90 inhibitor according to another embodiment. When the HSP90 inhibitor has a primary or secondary alcoholic hydroxyl group in the molecule, an aspartic acid derivative is used as a linker between the HSP90 inhibitor and the side chain carboxy group of the polyglutamic acid segment. A method of bonding can be mentioned. That is, the aspartic acid derivative serving as a linker is a derivative in which one carboxy group is amidated, and the amino group of the aspartic acid derivative and the side chain carboxy group of the polyglutamic acid segment of the block copolymer are bonded by an amide bond. A bonded group. The HSP90 inhibitor can be bound to the polyglutamic acid segment by ester-linking the remaining carboxy group of this aspartic acid derivative and the alcoholic hydroxyl group of the HSP90 inhibitor. When the HSP90 inhibitor conjugate using the aspartic acid derivative as a linker is administered in vivo, the HSP90 inhibitor dissociates non-enzymatically, and the drug can be released slowly.

［式中、Ｒ_８、Ｒ_９はそれぞれ独立して水素原子または（Ｃ１〜Ｃ６）アルキル基を示し、Ｒ_１０は水素原子、（Ｃ１〜Ｃ４０）アルキル基、（Ｃ１〜Ｃ４０）アラルキル基、芳香族基及びカルボキシ基が保護されたアミノ酸残基からなる群から選択される１種以上の置換基を示し、ＣＸ−ＣＹはＣＨ−ＣＨ若しくはＣ＝Ｃ（二重結合）を示し、ＤはＨＳＰ９０阻害剤の結合残基を示す。］で表される基が挙げられる。すなわち、本発明においてリンカーとするアスパラギン酸誘導体としては、一般式（３）または一般式（４）で表される基であることが好ましい。ここで、ＣＸ−ＣＹがＣ＝Ｃ（二重結合）の場合はＥ配置が好ましい。また、ＤがＨＳＰ９０阻害剤の結合残基であり、アスパラギン酸誘導体の末端カルボキシル基とＨＳＰ９０阻害剤とがエステル結合により結合している態様が好ましい。When R ₃ is an aspartic acid residue to which an HSP90 inhibitor is bound, examples of the aspartic acid residue include the following general formula (3) or general formula (4).

[Wherein R ₈ and R ₉ each independently represent a hydrogen atom or a (C1 to C6) alkyl group, and R ₁₀ represents a hydrogen atom, a (C1 to C40) alkyl group, a (C1 to C40) aralkyl group, an aromatic group. One or more substituents selected from the group consisting of amino acid residues in which a group group and a carboxy group are protected, CX-CY represents CH-CH or C = C (double bond), and D represents HSP90. The binding residues of the inhibitor are indicated. ] Is represented. That is, the aspartic acid derivative used as a linker in the present invention is preferably a group represented by the general formula (3) or the general formula (4). Here, when CX-CY is C = C (double bond), the E configuration is preferable. Further, it is preferable that D is a binding residue of the HSP90 inhibitor, and the terminal carboxyl group of the aspartic acid derivative and the HSP90 inhibitor are bonded by an ester bond.

Examples of the (C1-C6) alkyl group in R ₈ and R ₉ include, for example, methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, s-butyl group, isobutyl group, n-pentyl group, n -A hexyl group etc. are mentioned. R ₈ and R ₉ are preferably hydrogen atoms.

The (C1-C40) alkyl group in R ₁₀ is a (C1-C40) alkyl group which may have a substituent, and examples thereof include a methyl group, an ethyl group, an n-propyl group, an isopropyl group, and n-butyl. Group, s-butyl group, isobutyl group, n-pentyl group, n-hexyl group, n-stearyl group and the like. Examples of the substituent include a phenyl group, a naphthyl group, a methoxy group, an ethoxy group, and a dimethylamino group.
The (C1-C40) aralkyl group is an optionally substituted (C1-C40) aralkyl group, and examples thereof include a benzyl group, a naphthylmethyl group, a phenethyl group, and a 4-phenylbutyl group. . Examples of the substituent include a methyl group, an ethyl group, a nitro group, a chlorine atom, a bromine atom, and a dimethylamino group.
The aromatic group is an aromatic group that may have a substituent, and examples thereof include a phenyl group, a 4-nitrophenyl group, a 4-chlorophenyl group, a naphthyl group, and a fluorenyl group.
In addition, the substitution position of the substituent in each group of R ₈ , R ₉ , and R ₁₀ is not particularly limited as long as substitution is possible, and the number of substituents is not particularly limited.

In addition, the amino acid of the amino acid residue in which the carboxy group in R ₁₀ is protected includes an amino acid in which the carboxy group used in normal peptide synthesis is protected, and the carboxy group of the amino acid is protected by an ester or an amide. Are preferred.
The amino acid protected with an ester is, for example, an (C1-C12) alkyl ester optionally having an amino acid substituent, for example, a (C1-C12) alkyl ester of alanine, α of aspartic acid or β- (C1-C12) alkyl ester, α- or γ- (C1-C12) alkyl ester of glutamic acid, (C1-C12) alkyl ester of phenylalanine, (C1-C12) alkyl ester of cysteine, (C1-C12 of glycine) ) Alkyl ester, (C1-C12) alkyl ester of leucine, (C1-C12) alkyl ester of isoleucine, (C1-C12) alkyl ester of histidine, (C1-C12) alkyl ester of proline, (C1-C12 of serine) ) Alkyl esters, (Cl -C 12) alkyl esters of threonine, valine (Cl -C 12) alkyl ester, tryptophan (Cl -C 12) alkyl esters, tyrosine (Cl -C 12) alkyl esters.
Particularly preferred are phenylalanine methyl ester, glycine methyl ester, glycine (4-phenyl-1-butanol) ester, leucine methyl ester, phenylalanine benzyl ester, phenylalanine (4-phenyl-1-butanol) ester and the like. The amino acid may be D-form, L-form or a mixture thereof.

R ₁₀ is particularly preferably an n-butyl group, a 4-phenylbutyl group, and the like, and among them, a phenylbutyl group is particularly preferable. Moreover, it is preferable that CX-CY is CH-CH.

［式中、Ｘ_１は直鎖若しくは分岐の（Ｃ１〜Ｃ６）アルキル基または−ＣＯＮＲ_１１Ｒ_１２を示し、前記Ｒ_１１及びＲ_１２は同一でも異なってもいてもよい直鎖状、分岐状または環状の（Ｃ１〜Ｃ６）アルキル基であり、Ｘ_２は置換基を有していてもよいアリール基を示す］で示される１，２，４−トリアゾール−３−オン置換レゾルシン誘導体を挙げることができる。なお、一般式（２）におけるＸ_１、Ｘ_２、Ｒ_１１、Ｒ_１２における置換基の定義は、前述と同義である。In General Formula (3) or General Formula (4), D is a binding residue of the HSP90 inhibitor. The embodiment in which the HSP90 inhibitor is bonded to the terminal carboxyl group of the aspartic acid derivative by an ester bond is preferable. Examples of the HSP90 inhibitor include several developed compounds such as geldanamycin derivatives and substituted resorcin derivatives.
As the HSP90 inhibitor used for D, a substituted resorcin derivative having a hydroxyl group that can be bonded to the polyglutamic acid segment by an ester bond is particularly preferable. Examples of the substituted resorcin derivative include those represented by the general formula (2)

[Wherein, X ₁ represents a linear or branched (C ₁ -C 6) alkyl group or —CONR ₁₁ R ₁₂ , and R ₁₁ and R ₁₂ may be the same or different, and may be linear, branched or A cyclic (C1 to C6) alkyl group, and X ₂ represents an aryl group which may have a substituent]], 1,2,4-triazol-3-one substituted resorcin derivatives it can. In addition, the definition of the substituent in X < ₁ >, X < ₂ >, R <_11> , R < ₁₂ > in General formula (2) is synonymous with the above-mentioned.

一般式（８）、（９）及び（１０）において、Ｒ_８、Ｒ_９、Ｒ_１０、ＣＸ−ＣＹは上記一般式（３）及び（４）において定義された置換基群と同義である置換基群から選ばれる置換基が好ましい。これらの置換基が、高分子化合物中に存在する場合にはこれらの基が混在していてもよい。
ここでＲ_８、Ｒ_９、Ｒ_１０、ＣＸ−ＣＹは前記Ｒ_３におけるＲ_８、Ｒ_９、Ｒ_１０、ＣＸ−ＣＹと同様であり、好ましい基も同様である。R ₄ in the general formula (1) is an aspartic acid residue represented by the following general formula (8) or general formula (9) and / or an aspartic imide residue represented by the following general formula (10).

In the general formulas (8), (9) and (10), R ₈ , R ₉ , R _{10 and} CX-CY are the same as the substituent groups defined in the general formulas (3) and (4). A substituent selected from the group of groups is preferred. When these substituents are present in the polymer compound, these groups may be mixed.
Wherein _{R 8, R 9, R 10} , CX-CY is the same as _{R 8, R 9, R 10} , CX-CY in the _{R 3,} it is the same preferable groups.

In the polyglutamic acid segment in the general formula (1), a glutamic acid moiety to which any of EHC, R ₃ , R ₄ and R ₅ is not bonded may be present. In the general formula (1), a glutamic acid residue having a free acid type carboxy group is shown, but the carboxy group may be an alkali metal salt or an alkaline earth metal salt, and a glutamic acid moiety containing such a carboxyl group is included. The polymer compound of the general formula (1) having the formula is also included in the present invention. Examples of the alkali metal salt or alkaline earth metal salt include lithium salt, sodium salt, potassium salt, magnesium salt, calcium salt and the like. Furthermore, when it is provided parenterally as an anticancer agent, it is prepared as a solution in a solution, but it may be a glutamate based on a salt of the pH buffer of the solution with a cation.

In the polyglutamic acid segment in the present invention, a glutamic acid unit (containing d is the number of glutamic acid units) in which EHC is bonded to the side chain carboxy group, and a glutamic acid unit (including the number of units containing the R ₃ is bonded to the side chain carboxy group). e), a glutamic acid unit in which R ₄ is bonded to the side chain carboxy group (containing f), and a glutamic acid unit in which R ₅ is bonded to the side chain carboxy group (containing unit is g). ), The side chain carboxy group has a glutamic acid unit (containing unit is h) which is a free carboxy group or a salt thereof. Among these, the glutamic acid unit to which EHC is bound and the glutamic acid unit to which R ₃ which is a binding residue of the HSP90 inhibitor is bound are essential constituent units. On the other hand, the glutamic acid unit to which R ₄ is bonded, the glutamic acid unit to which R ₅ is bonded, and the glutamic acid unit which is a free carboxy group or a salt thereof are arbitrary constituent units. Each of these glutamic acid units is independently and randomly arranged.

The total number of glutamic acid units in the polyglutamic acid segment of the general formula (1) is represented by d + e + f + g + h. The ratio of d is 5 to 50%, preferably 5 to 40%, the ratio of e is 5 to 90%, preferably 5 to 80%, and the ratio of f is 0 to 90%, preferably 0 to 60%. The proportion is 0 to 90%, preferably 5 to 40%, and the proportion of h is 0 to 90%, preferably 0 to 30%.
In the present invention, the ratio of the drug content of EHC, which is a camptothecin derivative, and an HSP90 inhibitor is not particularly limited, and can be arbitrarily set at a mixture ratio that provides a synergistic effect enhancing action by the combined use of EHC and HSP90 inhibitor. can do. In the present invention, the mixing ratio of EHC and HSP90 inhibitor is indicated by the amount of each drug bound in the polyglutamic acid segment, and is set by the d value and e value representing the ratio of each unit of the polyglutamic acid segment. In the polymer compound of the present invention, it is considered that EHC and HSP90 inhibitor are released from the block copolymer and have a medicinal effect. For this reason, the binding amount of each drug should be set to a combined concentration ratio that exhibits a synergistic effect enhancing action in consideration of the dissociation rate of the drug from the block copolymer. Preferably, the mixing ratio of EHC and HSP90 inhibitor, d: e, is in the range of d: e = 1: 0.5-15. More preferably, it is 1: 1-10.
When the 1,2,4-triazol-3-one substituted resorcin derivative represented by the general formula (2) is used as the HSP90 inhibitor, a mixture of EHC and 1,2,4-triazol-3-one substituted resorcin derivative The ratio d: e is in the range of d: e = 1: 0.5 to 15 and more preferably d: e = 1: 1 to 10. In the case where a compound selected from the group consisting of general formula (5); compound 5, general formula (6); compound 6 and general formula (7); compound 7 is used as an HSP90 inhibitor, d: e Is in the range of d: e = 1: 0.5 to 15, more preferably d: e = 1: 1 to 10.

The polymer compound represented by the general formula (1) of the present invention may form a micelle having a polyethylene glycol segment as an outer shell and a polyglutamic acid segment to which a drug or the like is bound as an inner shell in water. That is, the polymer compound of the present invention is an antitumor agent using a block copolymer in which polyethylene glycol and polyglutamic acid are linked as a carrier, but becomes a polymer having a higher molecular weight due to the formation of a self-aggregate. The self-aggregate is preferably a high molecular weight body having a molecular weight of about 10 ^{6, and} has a pharmacokinetics depending on the molecular weight. That is, the drug is expected to have desirable pharmacokinetic properties for improving the efficacy of antitumor agents such as an increase in half-life, improved permeability to tumor tissues, and accumulation in tumor tissues.

  Next, a method for producing the polymer compound of the present invention will be described. In addition, the manufacturing method of the high molecular compound of this invention is not limited to the manufacturing method described here, and the method as described in the Example and reference example of a postscript.

  The method for constructing the main chain of the block copolymer having a polyethylene glycol segment and a polyglutamic acid segment, which is a polymer compound of the present invention represented by the general formula (1), is a method of binding a polyethylene glycol segment and a polyglutamic acid segment, Any method of sequentially polymerizing glutamic acid structural units to polyethylene glycol segments may be used.

As the latter method, in accordance with the method described in International Publication No. 2006/120914 pamphlet or the like, for example, polyethylene glycol modified with a modifying group such as a methyl group at one end and an aminopropyl group at the other end, N -The method of constructing a polyglutamic acid segment by sequentially reacting carbonyl glutamic anhydride. In this case, the N-carbonylglutamic acid anhydride may be a derivative of EHC in which the carboxy group of the glutamic acid side chain may be modified with an appropriate carboxylic acid protecting group, and is a derivative having a predetermined carboxy group substituent in advance. Conjugates, conjugates of HSP90 inhibitors according to R ₃ of general formula (1), conjugates of R ₄ of general formula (1), conjugates of R ₅ of general formula (1), free carboxylic acids, etc. There may be. As the N-carbonylglutamic acid anhydride, it is preferable to use an N-carbonylglutamic acid anhydride whose side chain carboxy group is modified with an appropriate carboxylic acid protecting group.
The carboxylic acid protecting group is not particularly limited, but an ester protecting group is preferable. That is, for the side chain carboxy group, an N-carbonylglutamic acid anhydride modified with an appropriate carboxylic acid protecting group is used, and the N-carbonylglutamic acid anhydride is reacted with a polyethylene glycol compound having an amino group modified at one end. Are sequentially polymerized to construct a block copolymer in which polyethylene glycol and a polyglutamic acid derivative are linked, to which EHC, an HSP90 inhibitor corresponding to R _{3 according} to the general formula (1), and a general formula (1 It is preferable to use a method of introducing substituents corresponding to R ₄ and R _{5 according} to). Prior to deprotection, the terminal amino group of the polyglutamic acid segment of the block copolymer may be acylated.

  More specifically, a polyethylene glycol compound modified with a methoxy group at one end and an amino group at the other end and γ-benzyl-N-carbonylglutamic anhydride are sequentially reacted, and then by subsequent polymerization, polyethylene glycol and poly (ethylene glycol) are reacted. A block copolymer linked with glutamic acid benzyl ester is prepared. Thereafter, by deprotecting the benzyl group of polyglutamic acid by a suitable method, a block copolymer in which the polyethylene glycol and polyglutamic acid are linked can be prepared. Examples of the deprotection reaction of the benzyl group include a hydrolysis reaction and a hydrogenation reduction reaction under alkaline conditions.

Next, a method for binding an EHC, HSP90 inhibitor or the like to the side chain carboxy group of the polyglutamic acid segment in the preparation of the polymer compound represented by the general formula (1) will be described.
A reaction in which EHC, an HSP90 inhibitor, an aspartic acid derivative to which an HSP90 inhibitor is bound, or the like is bound to the block copolymer in which the polyethylene glycol segment and the polyglutamic acid segment are linked is performed. The binding method is not particularly limited, and the ESP is bound first, and then the HSP90 inhibitor or the aspartic acid derivative to which the HSP90 inhibitor is bound may be bound simultaneously or in the reverse order. Also good.
Preferably, a block copolymer in which a polyethylene glycol segment and a polyglutamic acid segment are linked to an aspartic acid derivative to which an EHC and HSP90 inhibitor or an HSP90 inhibitor is bonded is subjected to a dehydration condensation reaction in the presence of a carbodiimide dehydration condensation agent. Good. According to this production method, a —N (R ₆ ) CONH (R ₇ ) group corresponding to R ₄ can be introduced into the block copolymer simultaneously with EHC and HSP90 inhibitors and the like.
As the carbodiimide dehydrating condensing agent, dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIPCI), 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (WSC) and the like can be used. In the dehydration condensation reaction, a reaction aid such as N, N-dimethylaminopyridine (DMAP) may be used.
In the present invention, the binding amount of EHC and HSP90 inhibitor can be adjusted by appropriately increasing or decreasing the amount of each drug charged in the dehydration condensation reaction. In addition, when dicyclohexyl carbodiimide (DCC) is used as a carbodiimide condensing agent, R ₆ and R ₇ of —N (R ₆ ) CONH (R ₇ ) are cyclohexyl groups. When a condensation reaction is performed using diisopropylcarbodiimide (DIPCI), R ₆ and R ₇ are isopropyl groups. When 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (WSC) is used, R ₆ and R ₇ of —N (R ₆ ) CONH (R ₇ ) are an ethyl group and 3-dimethylaminopropyl. It becomes a mixed substitution product.
After completion of the reaction, the polymer compound of the present invention can be produced via an optional purification step.

The polymer compound of the present invention gradually dissociates and releases EHC and HSP90 inhibitors, which are antitumor active ingredients, in a phosphate buffered saline (PBS) solution. This indicates that even when administered in vivo, it has physical properties to release the EHC and HSP90 inhibitors gradually.
It is known that high molecular weight compounds and low molecular weight compounds have greatly different pharmacokinetic behaviors in the body and their distribution in the body. From this, the high molecular compound of the present invention is different in drug expression characteristics and side effect expression characteristics from low molecular weight drugs, and can provide a new therapeutic method using a camptothecin derivative and an HSP90 inhibitor.

  The polymer compound of the present invention can be used as an anticancer agent, and the use of the anticancer agent is also included in the present invention. The anticancer agent can be used in commonly used preparations such as injections, drops, tablets, capsules, powders and the like. For formulation, commonly used pharmaceutically acceptable carriers and additives such as binders, lubricants, disintegrants, solvents, excipients, solubilizers, dispersants, stabilizers, suspensions Agents, preservatives, soothing agents, pigments, fragrances and the like can be used. Among them, it is preferably used as an injection or a drip infusion, water, physiological saline, 5% glucose or mannitol solution, water-soluble organic solvent (for example, glycerol, ethanol, dimethyl sulfoxide, N-methylpyrrolidone, polyethylene glycol, cremophor, etc. And a mixture thereof, and a mixture of water and the water-soluble organic solvent are preferred.

The dose of the polymer compound represented by the general formula (1) is appropriately set in consideration of the patient's sex, age, physique, physiological condition, disease state, therapeutic effect and the like. For example, the dose per body surface area of a patient is 0.01 to 500 mg / m ² , preferably 1 to 200 mg / m ² parenterally, in terms of EHC, which is an active ingredient, per day for an adult. Moreover, in terms of HSP90 inhibitor, the dose per patient body surface area is 0.01 to 500 mg / m ² , preferably 1 to 200 mg / m ² .

  The polymer compound of the present invention can be used in combination with other antitumor agents. Other antitumor agents are not particularly limited, and pharmaceuticals approved as antitumor agents and malignant tumor therapeutic agents can be used. That is, alkylating agents such as cyclophosphamide, ifosfamide and mitomycin C, platinum complexes such as cisplatin, carboplatin and oxaliplatin, anthracycline antitumor agents such as doxorubicin, epirubicin, pirarubicin and amrubicin, etoposide, etoposide phosphate, teniposide Etoposides, paclitaxel, docetaxel and other taxanes, vincristine, vinblastine, vindesine, vinca albinoids such as vinorelbine, 5-fluorouracil, tegafur, tegafur / uracil combination (UFT), tegafur / gimeracil / oteracil calcium (TS-1), nucleic acid antimetabolites such as flutulon, capecitabine, gemcitabine, cytosine arabinoside, methotrexate, pemetrexed Antifolate antifolates, steroids such as prednisolone and dexamethasone, anti-EGFR antibodies such as trastuzumab and cetuximab, EGFR tyrosine kinase inhibitors such as erlotinib, gefitinib and lapatinib, VEGFR tyrosine kinases such as sorafenib and sunitinib, everolimus and temsirolimus An anti-VEGF antibody such as an mTOR inhibitor or bevacizumab can be used.

  The polymer compound of the present invention is used for the treatment of malignant tumor diseases. The malignant tumor applied to the treatment with the polymer compound of 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 carcinoma, Prostate cancer, hepatocellular carcinoma, gastric cancer, pancreatic cancer, soft tissue sarcoma, Kaposi sarcoma, carcinoid carcinoma, head and neck cancer, melanoma, ovarian cancer, bile duct cancer, mesothelioma, and multiple myeloma, etc. widely It can be applied to general cancer treatment. In particular, non-small cell lung cancer, cervical cancer, ovarian cancer, gastric cancer (inoperable or recurrent), colorectal cancer (inoperable or recurrent), breast cancer (inoperable or recurrent) for which camptothecin derivatives are being treated Suitable for the treatment of squamous cell carcinoma, malignant lymphoma (non-Hodgkin lymphoma). In particular, high therapeutic effects can be expected in chemotherapy for various cancers that have undergone pretreatment with a camptothecin derivative and have decreased sensitivity to the camptothecin derivative in the pretreatment.

The present invention will be further described below with reference to examples. However, the present invention is not limited to these examples. The present invention products Gaussian distribution analysis showing the size of the particles constituting in aqueous solution (particle size) were performed at ^{Zeta Potential / Particlesizer NICOMP TM 380ZLS (} Particle Sizing Systems , Inc.).

[Synthesis Example 1]
5- (2,4-dihydroxy-5-isopropylphenyl) -4- (1-methyl-1H-indol-5-yl) -2,4-dihydro- [1,2,4] triazol-3-one ( Synthesis of Compound 5 (Compound represented by General Formula (5))) Compound 5 was synthesized based on the description of International Publication WO2007 / 13952. ¹ H-NMR of the obtained compound 5 is shown below.
¹ H-NMR (400 MHz, DMSO-d ₆ , ppm): 11.841 (s, 1H), 9.507 (s, 1H), 9.407 (s, 1H), 7.417-7.366 ( m, 3H), 6.921 (dd, 1H), 6.761 (s, 1H), 6.406 (dd, 1H), 6.205 (s, 1H), 3.777 (s, 3H), 2.911 (hept, 1H), 0.866 (d, 6H)

[Example 1]
Polymer compound in which compound 5 is bonded as EHC and R ₃ (compound 1: in general formula (1), R ₁ = methyl group, R ₂ = acetyl group, A = trimethylene group, no substituent corresponding to R ₄ ( f = 0), R ₆ = R ₇ = Isopropyl group, d + e + g + h = 23, t = 273, d + e + g + h is 10%, e is 35%, g is 20%, h is 35 The methoxypolyethylene glycol-polyglutamic acid block copolymer (with a molecular weight of 12,000 at one end is a methyl group and the other end is at aminopropyl) A methoxypolyethylene glycol segment as a group and a polyglutamic acid segment (terminal N acetyl group) having a polymerization number of 23. Binding groups was prepared as block copolymers) trimethylene group.
621 mg of the methoxypolyethylene glycol-polyglutamic acid block copolymer, 38 mg of 7-ethyl-10-hydroxycamptothecin (EHC) and 180 mg of the compound 5 obtained in Synthesis Example 1 were dissolved in 11.2 mL of DMF, and the mixture was dissolved at 35 ° C. for 15 minutes. After stirring, the mixture was stirred at 20 ° C. for 1 hour. Thereafter, 267 μL of diisopropylcarbodiimide (DIPCI) and 10.9 mg of N, N-dimethylaminopyridine (DMAP) were added, and the mixture was stirred at 20 ° C. for 21 hours. To this, 134 μL of DIPCI was added, and the mixture was further stirred for 2 hours. The reaction solution was slowly added dropwise to a mixed solution of 11.2 mL of ethyl acetate, 11.2 mL of ethanol and 90 mL of diisopropyl ether and stirred at room temperature for 1 hour. The precipitate was collected by filtration and washed with ethyl acetate. The obtained precipitate was dissolved in 46 mL of acetonitrile and 4.6 mL of water, added with an ion exchange resin (Dow Chemical Dowex 50 (H ⁺ ), 9.2 mL), stirred and filtered. Acetonitrile in the obtained filtrate was distilled off under reduced pressure, and then lyophilized to obtain Compound 1 (681 mg).

The content of EHC and compound 5 bound to compound 1 was analyzed by HPLC (high performance liquid chromatography) after adding 1N sodium hydroxide aqueous solution to compound 1 and stirring for 1 hour at 37 ° C. And calculated from the amount obtained from the calibration curve obtained with EHC and Compound 5. As a result, the contents of bound EHC and compound 5 were 3.4% (w / w) and 15.4% (w / w), respectively. Further, by ¹ H-NMR method, the bond content of —N (R ₆ ) CONH (R ₇ ) (R ₆ and R ₇ are isopropyl groups) corresponding to R ₅ according to the general formula (1) is 3.0 % (W / w).
When Gaussian distribution analysis was performed using an aqueous solution of Compound 1 (1 mg / mL), the obtained numerical value was 22 nm (volume weighting). Therefore, it was considered that Compound 1 self-associates in water to form a micelle-like aggregate.

[Example 2]
Polymer compound in which EHC and compound 5 are combined (compound 2: in general formula (1), R ₁ = methyl group, R ₂ = acetyl group, A = trimethylene group, and there is no substituent corresponding to R ₄ (f = 0 ), R ₆ = R ₇ = Isopropyl group, d + e + g + h = 23, t = 273, d + e + g + h is 30%, e is 20%, g is 20%, h is 30%) Synthesis A methoxypolyethyleneglycol-polyglutamic acid block copolymer (methoxyl having a molecular weight of 12,000 on one end with a methyl group and the other end with an aminopropyl group) by a method according to the method described in International Publication No. 2006/120914 It consists of a polyethylene glycol segment and a polyglutamic acid segment (terminal N acetyl group) having a polymerization number of 23, and the binding group is A block copolymer which is a rimethylene group) was prepared.
The methoxypolyethylene glycol-polyglutamic acid block copolymer 389 mg, EHC 70 mg, and Compound 5 70 mg obtained in Synthesis Example 1 were dissolved in 7.0 mL of DMF, stirred at 35 ° C. for 30 minutes, and then stirred at 20 ° C. for 2 hours. did. To this was added 167 μL of DIPCI and 6.9 mg of DMAP, and the mixture was stirred at 20 ° C. for 21 hours. Thereafter, 84 μL of DIPCI was added, and the mixture was further stirred for 2 hours. The reaction solution was slowly added dropwise to a mixed solution of 7 mL of ethyl acetate, 7 mL of ethanol and 56 mL of diisopropyl ether, and stirred at room temperature for 1 hour. The precipitate was collected by filtration and washed with ethyl acetate. The resulting precipitate was dissolved in 29 mL of acetonitrile and 2.9 mL of water, ion exchange resin (Dow Chemical Dowex 50 (H ⁺ ), 7.6 mL) was added, and the mixture was stirred and filtered. Acetonitrile in the obtained filtrate was distilled off under reduced pressure, and then lyophilized to obtain Compound 2 (450 mg).

The content of EHC and compound 5 bound to compound 2 was determined by adding 1N sodium hydroxide aqueous solution to compound 1 and stirring at 37 ° C. for 1 hour, and then measuring the content of free EHC and compound 5 by HPLC (high performance liquid chromatography). And calculated from the amount obtained from the calibration curve obtained with EHC and Compound 5. As a result, the contents of bound EHC and compound 5 were 10.9% (w / w) and 9.0% (w / w), respectively. In addition, the bond content of —N (R ₆ ) CONH (R ₇ ) (where R ₆ and R ₇ are isopropyl groups) corresponding to R ₅ according to the general formula (1) is 2.9% by ¹ H-NMR method. (W / w).
When Gaussian distribution analysis was performed using an aqueous solution of Compound 2 (1 mg / mL), the obtained value was 29 nm (volume weighting). Therefore, it was considered that Compound 2 self-associates in water to form a micelle-like aggregate.

[Example 3]
Polymer compound in which EHC and compound 5 are combined (compound 3: in general formula (1), R ₁ = methyl group, R ₂ is acetyl group, A = propylene group, and there is no substituent corresponding to R ₄ (f = 0 ), R ₆ = R ₇ = isopropyl group, d + e + g + h = 10, t = 114, d + e + g + h ratio of d is 10%, e ratio is 40%, g ratio is 20%, h ratio is 30%) Synthesis A methoxypolyethyleneglycol-polyglutamic acid block copolymer (polyethylene having a molecular weight of 5,000 at one end with a methyl group and the other end with an aminopropyl group) by a method according to the method described in the pamphlet of International Publication No. 2006/120914 A block copolymer comprising a glycol segment and a polyglutamic acid segment (terminal acetyl group) having a polymerization number of 10 .
The methoxypolyethylene glycol-polyglutamic acid block copolymer 473 mg, EHC 32 mg, and Compound 5 150 mg obtained in Synthesis Example 1 were dissolved in DMF 10 mL, stirred at 35 ° C. for 25 minutes, and then stirred at 25 ° C. for 2 hours. Then, 235 μL of DIPCI and 9 mg of DMAP were added and stirred at 25 ° C. for 20 hours. Thereafter, 118 μL of DIPCI was added, and the mixture was further stirred for 5 hours. The reaction solution was slowly added dropwise to a mixed solution of 30 mL of ethyl acetate and 120 mL of diisopropyl ether, and stirred at room temperature for 2 hours. The precipitate was collected by filtration and washed with ethyl acetate. The obtained precipitate was dissolved in 11.0 mL of acetonitrile and 2.0 mL of water, an ion exchange resin (Dow Chemical Dowex 50 (H ⁺ ), 5.0 mL) was added, and the mixture was stirred and filtered. The obtained filtrate was transferred to a dialysis membrane and dialyzed overnight at 5 ° C., and then the solution in the dialysis membrane was recovered. Acetonitrile in the obtained solution was distilled off under reduced pressure, and then freeze-dried to obtain Compound 3 (500 mg).

The content of EHC and compound 5 bound to compound 3 was determined by adding 1N sodium hydroxide aqueous solution to compound 1 and stirring at 37 ° C. for 1 hour, and then measuring the content of free EHC and compound 5 by HPLC (high performance liquid chromatography). And calculated from the amount obtained from the calibration curve obtained with EHC and Compound 5. As a result, the contents of bound EHC and compound 5 were 3.6% (w / w) and 16.6% (w / w), respectively. In addition, the bond content of —N (R ₆ ) CONH (R ₇ ) (R ₆ and R ₇ are isopropyl groups) corresponding to R ₅ according to the general formula (1) is 3.4% by ¹ H-NMR method. (W / w).
When Gaussian distribution analysis was performed using an aqueous solution of Compound 3 (1 mg / mL), the obtained value was 21 nm (volume weighting). Therefore, it was considered that Compound 3 self-associates in water to form a micelle-like aggregate.

[Synthesis Example 2]
EHC-linked polyethylene glycol-polyglutamic acid block copolymer (control drug 1; in general formula (1), R ₁ = methyl group, R ₂ = acetyl group, A = trimethylene group, no substituent corresponding to R ₃ (e = 0), no substituent corresponding to R ₄ (f = 0), R ₆ = R ₇ = isopropyl group, d + g + h = 23, t = 273, the ratio of d to d + g + h is 50%, the ratio of g is 40 %, The ratio of h is 10%) Based on the description of International Publication WO 2004/39869, a methoxypolyethylene glycol-polyglutamic acid block copolymer (one end of a molecular weight of 12,000 is a methyl group and the other end is aminopropyl) Methoxypolyethyleneglycol structure part which is a group and a polyglutamic acid structure part having a polymerization number of 23 having an N-terminal modified with an acetyl group A block copolymer in which the bonding group is a trimethylene group) and 7-ethyl-10-hydroxycamptothecin (EHC) are reacted in the presence of diisopropylcarbodiimide (DIPCI) and N, N-dimethylaminopyridine (DMAP), Subsequently, the control medicine 1 was obtained by processing by ion-exchange resin (Dow Chemical Dowex 50 (H ^<+> )).
The obtained control drug 1 was hydrolyzed at 1N-sodium hydroxide aqueous solution at room temperature for 10 minutes, and then the liberated EHC was quantitatively analyzed by the HPLC method, and the EHC content was determined to be 21% (w / w )Met. In addition, the bond content of —N (R ₆ ) CONH (R ₇ ) (R ₆ and R ₇ are isopropyl groups) corresponding to R ₅ according to the general formula (1) by ¹ H-NMR method is 5.6%. (W / w).

[Synthesis Example 3]
HSP90 inhibitor-conjugated polyethylene glycol-polyglutamic acid block copolymer (control drug 2; in general formula (1), R ₁ = methyl group, R ₂ = acetyl group, A = trimethylene group, no EHC-bound glutamic acid unit (d = 0), R ₃ is a binding residue of compound 5 represented by the general formula (5), there is no substituent corresponding to R ₄ (f = 0), R ₆ = R ₇ = isopropyl group, d + g + h = 23 , T = 273, e + g + h synthesis 40%, g 20%, h 40%) Methoxypolyethylene glycol prepared according to the method described in International Publication WO 2006/120914 Pamphlet -Polyglutamic acid block copolymer (methoxy having a molecular weight of 12,000, one end is a methyl group and the other end is an aminopropyl group 519 mg of a block copolymer in which a polyethylene glycol segment and a polyglutamic acid segment having a polymerization number of 23 (terminal N acetyl group) and a linking group is a trimethylene group) Compound 5 obtained by Synthesis Example 1 (Formula (5)) 150 mg) was dissolved in 10 mL of DMF, stirred at 35 ° C. for 15 minutes, and then stirred at 25 ° C. for 1 hour. Thereafter, 234 μL of diisopropylcarbodiimide (DIPCI) and 13.7 mg of N, N-dimethylaminopyridine (DMAP) were added and stirred at 25 ° C. for 16 hours. To this, 117 μL of DIPCI was added, and the mixture was further stirred for 4 hours. The reaction solution was slowly added dropwise to a mixed solution of 20 mL of ethanol and 80 mL of diisopropyl ether, stirred at room temperature for 1 hour, and the precipitate was collected by filtration and washed with diisopropyl ether. The resulting precipitate was dissolved in 5.0 mL of acetonitrile and 5.0 mL of water, and then an ion exchange resin (Dow Chemical Dowex 50 (H ⁺ ), 2.0 mL) was added and stirred and filtered. Acetonitrile in the obtained filtrate was distilled off under reduced pressure, and then freeze-dried to obtain Control 2 (525 mg).
The content of Compound 5 bound to Control Drug 2 was analyzed by HPLC (high performance liquid chromatography) after adding 1N sodium hydroxide aqueous solution to Control Drug 2 and stirring for 1 hour at 37 ° C. The amount calculated from the calibration curve obtained with Compound 5 was calculated. As a result, the content of bound compound 5 was 17.2% (w / w). Moreover, the bond content of the group which is —N (R ₆ ) CONH (R ₇ ) (R ₆ and R ₇ are isopropyl groups) corresponding to R ₅ according to the general formula (1) by ¹ H-NMR method is 3 0.0% (w / w).

[Test Example 1] Release of EHC and Compound 5 from Compounds 1 to 3 in the absence of enzyme Compound 1, Compound 2 and Compound 3 were each 1 mg / mL in PBS (phosphate buffered saline; pH 7.1). And was incubated at 37 ° C. The amount of EHC and compound 5 released from each compound into the PBS solution was analyzed over time by HPLC and quantified using a standard curve. The amounts of EHC and compound 5 detected in the solution are shown in FIGS. 1 to 3 as a ratio with respect to the total drug amount determined from the drug contents of compounds 1 to 3.

  As is apparent from FIGS. 1 to 3, the polymer compounds (compounds 1 to 3) of the present invention dissociate and release both EHC and compound 5 slowly in a PBS solution free from hydrolase. I was able to. The drug release rates of EHC and Compound 5 varied depending on the composition of the block copolymer in which polyethylene glycol and polyglutamic acid were linked and the amount of each drug bound. It has been shown that the release rate of EHC and compound 5 can be controlled.

[Test Example 2] Antitumor effect test Mouse colon cancer Colon 38 subcultured subcutaneously in mice was made into a block of about 2 mm square, and transplanted subcutaneously on the dorsal side of hairy mice using a trocar. On the 16th day after tumor transplantation, the polymer compound (Compound 1) of the present invention, the control drug 1 and the control drug 2 were administered once intravenously at the doses shown in Table 1. Compound 1 and Controls 1 and 2 were used after being dissolved in a 5% glucose solution or physiological saline.

Tumor volume after the start of administration and after the start of administration, the major axis (L (mm)) and minor axis (W (mm)) of the tumor were measured over time with calipers, and the formula (L × W ² ) / 2 was used. The tumor volume was calculated, and the change in relative tumor volume with respect to the tumor volume on the administration start date in each administration group is shown in FIG.

  As shown in FIG. 4, no antitumor activity was observed in Control Drug 2 to which a 1,2,4-triazol-3-one substituted resorcin derivative having HSP90 inhibitory activity was bound. On the other hand, in the control drug 1 to which EHC, which is a camptothecin derivative, was bound, a tumor shrinking effect was observed until the fifth day of administration, but thereafter tumor regrowth was observed. On the other hand, when Compound 1, which is the polymer compound of the present invention, was administered, a high tumor reduction effect was observed over a long period of time despite single administration. From the above results, it was shown that the polymer compound of the present invention can be an antitumor agent exhibiting two characteristics of a strong tumor shrinking activity and a tumor growth inhibitory activity exerted over a long period of time.

Claims (4)

A method for producing a polymer compound in which a camptothecin derivative and an HSP90 inhibitor are bound to a block copolymer in which a polyethylene glycol segment and a polyglutamic acid segment are linked, wherein the polymer compound has the general formula (1)

[Wherein R ₁ represents a hydrogen atom or an optionally substituted (C1-C4) alkyl group, t represents an integer of 45-450, and A represents a (C1-C6) alkylene group. The ratio of d to e is d: e = 1: 1 to 10 , d + e + f + g + h represents an integer of 6 to 60, the ratio of d to d + e + f + g + h is 5 to 50%, the ratio of e is 5 to 90%, f 0 to 90%, g to 0 to 90%, h to 0 to 90%, R ₂ represents a hydrogen atom or a (C1 to C4) acyl group, and R ₃ represents an HSP90 inhibitor. Or an aspartic acid residue to which an HSP90 inhibitor is bound, wherein the HSP90 inhibitor has the general formula (2)

[Wherein, X ₁ represents a linear or branched (C1 to C6) alkyl group or —CONR ₁₁ R ₁₂ , and R ₁₁ and R ₁₂ may be the same or different (C1 to C6). A linear, branched or cyclic alkyl group, and X ₂ represents an aryl group which may have a substituent. Wherein R ₄ is an aspartic acid residue or an aspartic acid imide, wherein the hydroxyl group of the resorcin moiety is an ester bond to an HSP90 inhibitor; Represents a residue, R ₅ represents —N (R ₆ ) CONH (R ₇ ), R ₆ and R ₇ may be the same or different, a branched or cyclic (C 3 -C 6) alkyl group or three A branched or straight chain (C1-C5) alkyl group optionally substituted with a secondary amino group, wherein the polyglutamic acid segment is a glutamic acid unit in which 7-ethyl-10-hydroxycamptothecin is bound to a side chain carboxy group; A glutamic acid unit having an R ₃ group bonded to a side chain carboxy group, a glutamic acid unit having an R ₄ group bonded to a side chain carboxy group, A glutamic acid unit in which an R ₅ group is bonded to a ruxoxy group and a glutamic acid unit in which a side-chain carboxy group is a free carboxy group are each independently a polyglutamic acid structure arranged randomly. ] Is a polymer compound represented by,
A method for producing a polymer compound, comprising reacting a block copolymer in which a polyethylene glycol segment and a polyglutamic acid segment are linked with a camptothecin derivative, an HSP90 inhibitor, or an aspartic acid derivative to which an HSP90 inhibitor is bound . R ₃ is an aspartic acid residue to which an HSP90 inhibitor is bound, and the aspartic acid residue is represented by general formula (3) or general formula (4)

[Wherein R ₈ and R ₉ each independently represent a hydrogen atom or a (C1-C6) alkyl group, and R ₁₀ represents a hydrogen atom, a (C1-C40) alkyl group, a (C1-C40) aralkyl group, an aromatic group. One or more substituents selected from the group consisting of amino acid residues in which a group group and a carboxy group are protected, CX-CY represents CH-CH or C = C (double bond), and D represents HSP90. The binding residues of the inhibitor are indicated. ] Der is,
The aspartic acid residue, the amino group of the aspartic acid derivative, side-chain carboxy group of the polyglutamic acid segment of the block copolymer, Ru coupling groups der linked by an amide bond, the manufacturing method according to claim 1. The HSP90 inhibitor of the binding residue of the HSP90 inhibitor of D is represented by the general formula (2)

[Wherein, X ₁ represents a linear or branched (C1-C6) alkyl group or —CONR ₁₁ R ₁₂ , and R ₁₁ and R ₁₂ may be the same or different, and may be a linear or branched group. or cyclic (C1 -C6) alkyl group, X ₂ represents an aryl group which may have a substituent. The production method according to claim 2, which is a 1,2,4-triazol-3-one-substituted resorcin derivative represented by formula (I), wherein the hydroxyl group of the resorcin moiety is a binding residue of an HSP90 inhibitor having an ester bond. R ₁ is a methyl group or ethyl group, A is an ethylene group or propylene group, R ₂ is an acetyl group or propionyl group, R ₆ and R ₇ are both cyclohexyl groups or both isopropyl groups, and the ratio of d to d + e + f + g + h is 5 to 5 The ratio of e is 5 to 80%, the ratio of f is 0 to 60%, the ratio of g is 5 to 40%, and the ratio of h is 0 to 30%. The production method according to item.

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