WO2010086964A1 - Combination therapy for treating cancer - Google Patents

Abstract

Disclosed is a combination therapy for potentiating the effect of inhibiting tumor growth and/or metastasis wherein an inhibitor of plasminogen activator inhibitor-1 is combined with a cytotoxic agent or a cell growth inhibitor.

Description

Combination therapy for cancer treatment

The present invention relates to a medicament comprising a combination of a plasminogen activator inhibitor-1 (PAI-1) inhibitor and a cytotoxic agent or cytostatic agent, particularly a medicament for inhibiting tumor growth and / or metastasis. Use of the combination for the manufacture of a medicament for inhibiting tumor growth and / or metastasis, a pharmaceutical kit comprising a PAI-1 inhibitor and a cytotoxic or cytostatic agent, in particular tumor growth and Pharmaceutical kit for suppressing metastasis, method of suppressing tumor growth and / or metastasis administered by combining PAI-1 inhibitor and cytotoxic agent or cytostatic agent, and PAI-1 inhibitor The present invention relates to a medicament for enhancing the effect of inhibiting tumor growth and / or metastasis of a cytotoxic agent or cytostatic agent as an active ingredient.

Major antitumor drugs used in the chemotherapy area of malignant tumors include fluorinated pyrimidine preparations, DNA topoisomerase inhibitors, taxane drugs, molecular targeted drugs, platinum drugs and anticancer antibiotics. It is done.
The mechanism of action related to anticancer activity varies, for example, 5-fluorouracil, a fluoropyrimidine preparation, a typical antitumor drug used in gastrointestinal cancer, inhibits the DNA synthesis pathway by thymidylate synthase Antitumor effect is manifested by things. The major DNA topoisomerase inhibitor irinotecan hydrochloride is a semisynthetic derivative of camptothecin. DNA synthesis is inhibited by SN-38, an active metabolite of irinotecan, inhibiting type I DNA topoisomerase. Taxane drugs are drugs that act on microtubules during cell division, and include docetaxel hydrate and paclitaxel. Typical examples of molecular target drugs include bevacizumab, which is a vascular endothelial growth factor (VEGF) inhibitor, and cetuximab, panituzumab, which are epidermal growth factor receptor (EGFR) inhibitors. Vascular endothelial growth factor is a secreted form of angiogenesis-inducing factor and plays a central role in the formation of tumor blood vessels. Epidermal growth factor receptors play an important role in the control of metabolic processes such as cell differentiation, proliferation, invasion, apoptosis and angiogenesis. Platinum-based drugs are drugs that specifically bind to DNA and exert an antitumor effect by inhibiting DNA replication during cell division. Examples of platinum-based drugs include cisplatin, nedaplatin, and oxaliplatin. Anticancer antibiotics include actinomycins, mitomycins, anthracyclines and bleomycins, which act as DNA or RNA inhibitors and inhibit the growth and proliferation of cancer cells.

However, these chemotherapeutic agents have useful anti-tumor effects, but bone marrow suppression, interstitial pneumonia, hair loss, heart failure, DIC, intestinal perforation, hair loss, peripheral neuropathy, cardiotoxicity, kidney damage, hearing and nerves Side effects such as disability, edema, shock, stomatitis, allergic symptoms, and hemolytic anemia often occur, and this is a great burden for patients in cancer treatment.
Reducing the side effects of these chemotherapeutic agents is important in the treatment of cancer. As one of the measures for reducing the side effects, there can be considered a combination therapy that enables the dose of a chemotherapeutic agent having a strong side effect to be reduced by improving the anticancer action by the combined effect. To date, various combination therapies that combine multiple anticancer agents have been tried.

For example, for gastrointestinal cancer chemotherapy, a combination of 5-fluorouracil and leucovorin and oxaliplatin, irinotecan and 5-fluorouracil and leucovorin, folic acid and 5-fluorouracil and irinotecan, irinotecan and folic acid and 5-fluorouracil, oxaliplatin and irinotecan Multi-drug combination therapy is performed by such means. In chemotherapy for lung cancer, combination therapy using an anticancer drug such as irinotecan, paclitaxel, docetaxel, vinorelbine, gemcitabine, amrubicin and a platinum-based drug for advanced non-small cell lung cancer is performed. In ovarian cancer chemotherapy, a combination of taxane taxol and a platinum preparation, and a combination of adriamycin and a platinum preparation cisplatin as a treatment for advanced / recurrent endometrial cancer are being studied. In the chemotherapy of bladder cancer and prostate cancer, combination therapy of methotrexate, vinblastine, adriamycin, and cisplatin and gemcitabine and cisplatin is being investigated for urothelial cancer.

In pancreatic cancer chemotherapy, gemcitabine is currently recognized worldwide as a first-line drug. Pancreatic cancer is one of the most difficult cancers to treat and its results are not satisfactory. Combination therapy with gemcitabine and other drugs has been actively attempted for patients with pancreatic cancer.
For example, many combinations such as combination therapy of 5-fluorouracil and gemcitabine (Non-patent Document 1), combination therapy of oxaliplatin and gemcitabine (Non-patent document 2), combination therapy of irinotecan hydrochloride and gemcitabine (Non-patent document 3) The results of trials with therapy have been reported, but none of them has been shown to have a significant increase in survival.

As a study on the combined effect of other combinations for cancer, for example, a report on the combined effect of a combination of a raf kinase inhibitor and irinotecan or gemcitabine (Patent Document 1), an EGFR inhibitor and irinotecan Report on combined use (Patent Document 2), Report on combined use of EGFR inhibitor and gemcitabine (Patent Document 3), Report on combined use of serine / threonine kinase PLK-1 inhibitor and irinotecan or gemcitabine (Patent Document 4), and NF There are reports on the combined use of a protease inhibitor having -κB inhibitory activity and gemcitabine (Patent Document 5), etc., all of which are under development.

Reports on the antitumor effects of PAI-1 inhibitors are disclosed in Non-Patent Document 4, Patent Documents 6-10, and the like. However, to date, there is no known document regarding the combined effect of a drug combining a PAI-1 inhibitor and an existing anticancer drug (cytotoxic agent or cytostatic agent). In the case of multi-drug combination, not only the useful effects of each drug are synergistically amplified, but it is also possible that minor effects such as toxicity and side effects increase. Therefore, the effectiveness and safety of each drug used as a concomitant drug cannot be predicted in advance from the effectiveness and safety of the drug. There is a need for drug development based on solid data on safety and efficacy in combination administration.
Journal of Clinical Oncology, 20 (15), 3270-3275 (2002) Journal of Clinical Oncology, 23 (15), 3509-3516 (2005) Journal of Clinical Oncology, 22 (18), 3776-3783 (2004) Carcinogenesis vol.29 no.4 p.824-829 (2008) International Publication No. 2003/047579 Pamphlet International Publication No. 2005/117877 Pamphlet International Publication No. 2005/117878 Pamphlet International Publication No. 2006/018182 Pamphlet International Publication No. 2007/013696 Pamphlet International Publication No. 2005/030750 Pamphlet International Publication No. 2006/023864 Pamphlet International Publication No. 2006/023865 Pamphlet International Publication No. 2006/023866 Pamphlet International Publication No. 2007/138705 Pamphlet

The problem to be solved by the present invention is to provide a combination therapy for suppressing the growth and / or metastasis of tumors by the combined use of individual drugs, in which the effectiveness and safety of each drug are secured and enhanced. There is.

The present inventors have conducted intensive research on a combination therapy combining an existing anticancer agent (cytotoxic agent or cytostatic agent) with another agent for the purpose of suppressing tumor growth and / or metastasis. As a result of overlapping, it has been found that an antitumor effect possessed by a cytotoxic agent or cytostatic agent is improved by an unprecedented combination of a PAI-1 inhibitor and a cytotoxic agent or cytostatic agent. It came to make.

Therefore, the present invention relates to a pharmaceutical comprising a combination of a plasminogen activator inhibitor-1 (PAI-1) inhibitor and a cytotoxic agent or cytostatic agent, particularly to suppress tumor growth and / or metastasis. It is related with the medicine for.
The present invention further relates to the use of a combination of a PAI-1 inhibitor and a cytotoxin or cytostatic agent for the manufacture of a medicament for inhibiting tumor growth and / or metastasis. .
Furthermore, the present invention relates to a pharmaceutical kit comprising a PAI-1 inhibitor and a cytotoxin or cytostatic agent, in particular, a pharmaceutical kit for suppressing tumor growth and / or metastasis.
Furthermore, the present invention relates to a method for suppressing tumor growth and / or metastasis, comprising administering a PAI-1 inhibitor and a cytotoxic agent or cytostatic agent to a mammal including a human.
Furthermore, the present invention relates to a medicament for enhancing the tumor growth and / or metastasis-suppressing effect of a cytotoxic agent or cytostatic agent containing a PAI-1 inhibitor as an active ingredient.

Combination therapy combining a PAI-1 inhibitor with a cytotoxic agent or cytostatic agent is an antitumor agent that is completely different from conventional combination therapy, and is used as a prophylactic, therapeutic agent for various malignant tumors, or for metastasis. It can be used as a prophylactic agent, and exhibits a high tumor growth inhibitory effect on pancreatic cancer cells, particularly in combination with gemcitabine. In addition, the combined effect can reduce the amount of cytotoxic agents and cytostatics having strong side effects, which is also effective from the viewpoint of the patient's quality of life (QOL).

〔式（１）において環Ａは、ベンゾオキサゾール、ベンゾチアゾール又はピリミジンを表し；Ｒ_１は、水酸基、炭素数１－６個の直鎖もしくは分岐状のアルキル基、炭素数１－４個の直鎖もしくは分岐状の低級アルコキシ基、シクロヘキシルメトキシ基、ベンジルオキシ基（ここで、ベンジルオキシ基のフェニルは、水酸基、炭素数１－４個の直鎖もしくは分岐状の低級アルキル基、炭素数１－４個の直鎖もしくは分岐状の低級アルコキシ基、トリフルオロメチル基、ニトロ基、－Ｎ（Ｒ_４、Ｒ_５）基（ここで、Ｒ_４、Ｒ_５は、それぞれ独立して水素原子、炭素数１－６個の直鎖もしくは分岐状のアルキル基又はベンジル基を意味する。）及びハロゲン原子から選ばれる置換基で置換されていてもよい。）、トリフルオロメチル基、ニトロ基、－Ｎ（Ｒ_４、Ｒ_５）基（ここで、Ｒ_４、Ｒ_５は、前記定義と同じである。）、ハロゲン原子、フェニル基（ここで、フェニル基は水酸基、炭素数１－４個の直鎖もしくは分岐状の低級アルキル基、炭素数１－４個の直鎖もしくは分岐状の低級アルコキシ基、トリフルオロメチル基、ニトロ基、－Ｎ（Ｒ_４、Ｒ_５）基（ここで、Ｒ_４、Ｒ_５は、前記定義と同じである。）及びハロゲン原子から選ばれる置換基で置換されていてもよい。）を表し；Ｒ_２及びＲ_３は、それぞれ独立して水素原子又は炭素数１－４個の直鎖もしくは分岐状の低級アルキル基を表し；ｍは、０又は１から４の整数を表し、ｍ個のＲ_１は、それぞれ同一又は異なってもよく；Ｗは、１から６の整数を表す。〕
で表されるプロパン二酸誘導体、又は薬学的に許容されるその塩が好ましい。 Hereinafter, the present invention will be described in detail.
The PAI-1 inhibitor used in the present invention includes the following formula (1)

[In the formula (1), ring A represents benzoxazole, benzothiazole or pyrimidine; R ₁ represents a hydroxyl group, a linear or branched alkyl group having 1 to 6 carbon atoms, and a straight chain having 1 to 4 carbon atoms. A chain or branched lower alkoxy group, a cyclohexylmethoxy group, a benzyloxy group (wherein phenyl of the benzyloxy group is a hydroxyl group, a linear or branched lower alkyl group having 1 to 4 carbon atoms, a 1- 4 linear or branched lower alkoxy groups, trifluoromethyl group, nitro group, —N (R ₄ , R ₅ ) group (where R ₄ and R ₅ are each independently hydrogen atom, carbon Represents a straight-chain or branched alkyl group or benzyl group having a number of 1-6, and may be substituted with a substituent selected from a halogen atom), a trifluoromethyl group, B group, -N _(R 4, _{R 5)} group _(wherein, R 4, _{R 5} are the same as defined above.), A halogen atom, a phenyl group (wherein the phenyl group is hydroxyl, C 1 -C -4 linear or branched lower alkyl groups, linear or branched lower alkoxy groups having 1 to 4 carbon atoms, trifluoromethyl group, nitro group, -N (R ₄ , R ₅ ) group ( Wherein R ₄ and R ₅ are the same as defined above) and may be substituted with a substituent selected from a halogen atom; and R ₂ and R ₃ are each independently hydrogen. Represents an atom or a linear or branched lower alkyl group having 1 to 4 carbon atoms; m represents 0 or an integer of 1 to 4, and m R _{1 s} may be the same or different; Represents an integer of 1 to 6. ]
Is preferred, or a pharmaceutically acceptable salt thereof.

Here, as the “linear or branched alkyl group having 1 to 6 carbon atoms”, specifically, methyl group, ethyl group, propyl group, isopropyl group, butyl group, isobutyl group, sec-butyl group, Examples thereof include a tert-butyl group, a pentyl group, an isopentyl group, a neopentyl group, a tert-pentyl group, a hexyl group, an isohexyl group, and a neohexyl group. Specific examples of the “linear or branched lower alkyl group having 1 to 4 carbon atoms” include methyl group, ethyl group, propyl group, isopropyl group, butyl group, isobutyl group, sec-butyl group, tert- A butyl group etc. are mentioned. Specific examples of the “C 1-4 straight or branched lower alkoxy group” include methoxy group, ethoxy group, propoxy group, isopropoxy group, butoxy group, isobutoxy group, sec-butoxy group, tert -Butoxy group and the like. A methoxy group and an ethoxy group are preferable. Specific examples of the “halogen atom” include a chlorine atom, a bromine atom, and a fluorine atom.

The propanedioic acid derivative represented by the formula (1) may be a pharmaceutically acceptable salt. As used herein, “pharmaceutically acceptable salt” refers to a salt prepared from a pharmaceutically acceptable non-toxic base or acid. As a PAI-1 inhibitor, the salt of the propanedioic acid derivative represented by the formula (1) preferably used in the present invention is prepared from a pharmaceutically acceptable non-toxic base including an inorganic base and an organic base. I can do things. Salts derived from inorganic bases include potassium salts, sodium salts, calcium salts, magnesium salts, aluminum salts, ammonium salts, and the like. Particularly preferred are potassium salt and sodium salt.
Salts derived from pharmaceutically acceptable organic non-toxic bases include primary amine salts, secondary amine salts, tertiary amine salts, cyclic amine salts and the like.
Further, when the structure of the propanedioic acid derivative represented by the formula (1) has a basic site, a salt can be prepared from a pharmaceutically acceptable non-toxic acid including an inorganic acid and an organic acid. . Examples of non-toxic acids include acetic acid, citric acid, succinic acid, tartaric acid, maleic acid, methane sulfonic acid, benzene sulfonic acid, toluene sulfonic acid, hydrochloric acid, sulfuric acid, phosphoric acid and the like.

In the present invention, the propanedioic acid derivative represented by the formula (1) that is preferably used as a PAI-1 inhibitor is disclosed in Non-Patent Document 4 and Patent Document 10, and described in Patent Document 10 It can be manufactured by a method.

In the present invention, as the PAI-1 inhibitor, in particular, ring A is benzoxazole or pyrimidine; R ₁ is a linear or branched alkyl group having ₁ to 6 carbon atoms, cyclohexylmethoxy group, benzyl An oxy group (wherein the phenyl of the benzyloxy group may be substituted with a halogen atom) or a phenyl group (wherein the phenyl group is a straight or branched lower alkoxy having 1 to 4 carbon atoms) R ₂ and R ₃ are hydrogen atoms; m is an integer of 1 or 2, and two R _{1 s} may be the same or different from each other; W is an integer of 3 to 5, and the propanedioic acid derivative represented by the formula (1) or a pharmaceutically acceptable salt thereof is more preferable.

Particularly preferred examples of the propanedioic acid derivative represented by the formula (1) include the following compounds.
(1). [3-[[6- (5-phenyl-2-benzoxazolyl) -2-naphthalenyl] oxy] propyl] propanedioic acid,
(2). [5-[[6- [5- (1,1-dimethylethyl) -2-benzoxazolyl] -2-naphthalenyl] oxy] pentyl] propanedioic acid,
(3). [5-[[6- [6- (cyclohexylmethoxy) -2-benzoxazolyl] -2-naphthalenyl] oxy] pentyl] propanedioic acid,
(4). [5-[[6- [6- (phenylmethoxy) -2-benzoxazolyl] -2-naphthalenyl] oxy] pentyl] propanedioic acid,
(5). [3-[[6- [4-[(2-fluorophenyl) methoxy] -6- (4-methoxyphenyl) -2-pyrimidinyl] -2-naphthalenyl] oxy] propyl] propanedioic acid,
(6). [5-[[6- [4-Phenyl-6- (phenylmethoxy) -2-pyrimidinyl] -2-naphthalenyl] oxy] pentyl] propanedioic acid,
(7). [5-[[6- [4-cyclohexylmethoxy-6- (1,1-dimethylethyl) -2-pyrimidinyl] -2-naphthalenyl] oxy] pentyl] propanedioic acid,
(8). [5-[[6- [4- (1,1-dimethylethyl) -6-[(4-fluorophenyl) methoxy] -2-pyrimidinyl] -2-naphthalenyl] oxy] pentyl] propanedioic acid and (9 ). [5-[[6- [4-[(2-Chlorophenyl) methoxy] -6- (1,1-dimethylethyl) -2-pyrimidinyl] -2-naphthalenyl] oxy] pentyl] propanedioic acid.

Table 1 shows the structural formulas of the above-described compounds.

Among these, in particular, [5-[[6- [4-phenyl-6- (phenylmethoxy) -2-pyrimidinyl] -2-naphthalenyl] oxy] pentyl] propanedioic acid, and [5-[[6 -[5- (1,1-dimethylethyl) -2-benzoxazolyl] -2-naphthalenyl] oxy] pentyl] propanedioic acid is preferred.

As used herein, “cytotoxic agent” refers to a drug administered to kill or kill cancer cells, and “cytostatic agent” refers to a drug administered to suppress tumor growth. . The cytotoxic agent or cytostatic agent used in the present invention may form a pharmaceutically acceptable non-toxic salt.

Preferred examples of cytotoxins or cytostatics include DNA synthesis inhibitors, DNA topoisomerase type I inhibitors, DNA topoisomerase type II inhibitors, microtubule agents, vascular endothelial growth factor (VEGF) inhibitors, epithelium Examples include growth factor receptor (EGFR) inhibitors, alkylating agents, platinum complexes, anticancer antibiotics, hormone therapy agents, etc. In the present invention, one or more of these are combined with PAI-1 inhibitors. Have been used.
More preferred specific examples of the cytotoxic agent or cytostatic agent in the present invention include gemcitabine, capecitabine, 5-fluorouracil, methotrexate, irinotecan, etoposide, doxorubicin, vinorelbine, paclitaxel, vincristine, vinblastine, docetaxel, bevacizumab, gefititib, Examples include trastuzumab, sorafenib, busulfan, melphalan, cyclophosphamide, cisplatin, carboplatin, oxaliplatin, mitomycin C, actinomycin D, tamoxifen, and anastrozole.

A preferred embodiment of the present invention includes a combination of a PAI-1 inhibitor and gemcitabine. Other preferred specific examples include a combination of a PAI-1 inhibitor and irinotecan.
Gemcitabine is metabolized in cells by deoxycytidine kinase to the active nucleotides diphosphate and triphosphate, and is thought to directly and indirectly inhibit DNA synthesis. Gemcitabine has broad antitumor activity against solid tumors such as pancreatic cancer, lung cancer, prostate cancer, ovarian cancer and breast cancer.
Irinotecan is believed to inhibit DNA synthesis by inhibiting DNA topoisomerase (type I), which performs single-strand breaks and recombination of DNA. That is, type I DNA topoisomerase binds to DNA and then cleaves one of the double-stranded DNA. At this time, irinotecan or its active metabolite SN-38 forms a complex with type I DNA topoisomerase. It is considered that recombination of DNA is inhibited by stabilizing the complex. Irinotecan has antitumor activity against solid tumors such as small cell lung cancer, non-small cell lung cancer, cervical cancer, ovarian cancer, gastric cancer and colorectal cancer.

In the present invention, tumor growth and / or metastasis can be effectively suppressed by a combination of a PAI-1 inhibitor and a cytotoxic agent or cytostatic agent.
The tumors targeted in the present invention include lung cancer, non-small cell lung cancer, breast cancer, pancreatic cancer, liver cancer, kidney cancer, bile duct cancer, esophageal cancer, stomach cancer, duodenal cancer, small intestine cancer, large intestine. (Colon, rectal) cancer, uterine cancer, ovarian cancer, bladder cancer, prostate cancer, thyroid cancer, parathyroid cancer, mesothelioma, leukemia, malignant lymphoma, melanoma, multiple myeloma Examples include malignant tumors and colon or rectal precancerous polyps, and precancerous lesions such as gastric epithelial malformations. Preferably, digestive tract cancer and digestive adenocarcinoma such as pancreatic cancer, liver cancer, bile duct cancer, esophageal cancer, stomach cancer, duodenal cancer, small intestine cancer, large intestine (colon, rectum) cancer It is said. Particularly preferred are pancreatic cancer and colon (colon, rectal) cancer.

In the present invention, the PAI-1 inhibitor and the cytotoxic agent or cytostatic agent may be administered simultaneously, or may be administered separately. When administering a PAI-1 inhibitor and a cytotoxic agent or cytostatic agent, it is administered in the form of a commonly used formulation together with a pharmaceutically acceptable carrier.
The pharmaceutically acceptable carrier can be a solid, liquid or gas, and the solid carrier includes gum arabic, agar, gelatin, talc, lactose and the like. Examples of the liquid carrier include water, olive oil, sugar syrup, and the like, and examples of the gas carrier include nitrogen.

PAI-1 inhibitors are usually administered to mammals including humans orally, parenterally, transdermally, by injection, by inhalation or spray, sublingually, rectally, Or it can be administered vaginally.

A PAI-1 inhibitor preparation for oral administration is prepared by adding a pharmacologically and pharmaceutically acceptable additive, etc., if necessary, together with the above-mentioned pharmaceutically acceptable carrier, and by a conventional method. Or it can manufacture as a capsule. Moreover, it can also be set as the tablet or capsule containing both a PAI-1 inhibitor and a cytotoxic agent or a cytostatic.
Oral preparations include, for example, magnesium metasilicate aluminate, magnesium silicate, magnesium carbonate, calcium hydrogenphosphate, various starches, excipients such as dextrin, ethylcellulose (EC), hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose ( Additives such as binders such as HPC), sodium alginate, gelatin, and corrigents such as mannitol, citric acid, sodium citrate, and sugar, and other additives such as disintegrants, wetting agents, and coating agents can be used. Oral solutions may be in the form of aqueous or oily suspensions, solutions, emulsions, syrups, etc., for example suspensions such as methylcellulose, carboxymethylcellulose, sodium alginate, gelatin and propylene glycol, sucrose or Sweeteners such as saccharin, or emulsifiers such as gelatin, macrogol (PEG), and propylene glycol fatty acid esters can be included.

Administration by injection means intravenous, intraarterial, intramuscular and subcutaneous injection. An injection of a PAI-1 inhibitor can be prepared as a sterile aqueous solution. Furthermore, it can be in the form of a sterile powder for preparation into an injectable solution. Moreover, it can also be set as the injection containing both a PAI-1 inhibitor and a cytotoxic agent or a cytostatic agent.
As a carrier for injection, for example, water can be used as a solubilizing agent, and ethanol, glycerol, propylene glycol or the like can be used as a solubilizing agent. In addition, pharmaceutical ingredients such as a pH adjuster, an isotonic agent, and a stabilizer can also be used.
Suitable forms for transdermal administration can be, for example, creams, ointments, lotions and the like.
It can also be made into suppositories for rectal administration. Suppositories should be based on an appropriate substance such as cacao butter, lauric fat, macrogol, glycerogelatin, sodium stearate or mixtures thereof, and if necessary, add emulsifiers, suspending agents, preservatives, etc. Can do.

In the present invention, a kit containing both a PAI-1 inhibitor and a cytotoxic agent or a cytostatic agent may be used. The kit can be provided in the form of a single container or a plurality of containers. In the kit, each of the PAI-1 inhibitor and the cytotoxic agent or cytostatic agent can be contained in the form of a preparation as described above.

The propanedioic acid derivative having the PAI-1 inhibitory activity represented by the formula (1) can be administered by any of the administration routes described above, and the dose is 0.1 to 300 mg / kg to the patient. Can be administered. Furthermore, the PAI-1 inhibitor can be administered one or more times per day.
Cytotoxic agents or cytostatic agents can be administered orally, rectally, by inhalation or by injection or infusion. Administration by injection means intravenous, intramuscular and subcutaneous. The dose of the cytotoxic agent or cytostatic agent may be administered at a dose that is usually employed, or may be administered at a dose lower than the dose.

The cytotoxin or cytostatic agent can be administered by any of the administration routes described above in combination with the PAI-1 inhibitor. The PAI-1 inhibitor can be administered simultaneously with the cytotoxic agent or cytostatic agent. This can be done by administering a single formulation containing both a PAI-1 inhibitor and a cytotoxic or cytostatic agent, or by co-administering separate formulations.
In addition, it is not always necessary to simultaneously administer the PAI-1 inhibitor and the cytotoxic agent or cytostatic agent, whereas the PAI-1 inhibitor is administered once or more per day, whereas the cytotoxic agent or cell proliferation The inhibitor can be administered repeatedly after a certain period of time.
Also, the PAI-1 inhibitor and cytotoxic agent or cytostatic agent may be administered by different routes. Cytotoxic agents or cytostatic agents may be administered parenterally and PAI-1 inhibitors may be administered orally. In this case as well, the administration conditions can be arbitrarily determined according to the prescribed formulation of the cytotoxic agent or cytostatic agent.

In the present invention, gemcitabine, which is one of the preferred examples of the cytotoxic agent used in combination with the PAI-1 inhibitor, is marketed as an injectable hydrochloride in the form of hydrochloride, and is administered to the patient by intravenous injection or an appropriate infusion method. To be administered. Another preferred example, irinotecan, is also marketed in hydrochloride as an injectable drug and is administered to patients by intravenous injection or other suitable infusion method.

In the present invention, a drug containing a PAI-1 inhibitor as an active ingredient can be used to enhance the effect of suppressing tumor growth and / or metastasis of a cytotoxin or cytostatic agent. In this case as well, the PAI-1 inhibitor can be administered simultaneously or separately with the cytotoxic agent or cytostatic agent in the above-mentioned preparation form, administration route and dosage.

Hereinafter, the present invention will be described in more detail with reference to test examples and the like, but the present invention is not limited to these test examples and the like.

Materials and Methods For animal testing, BALB / cAJc1-nu, SPF, female mice (CLEA Japan, Inc.) were used. The mouse is a strain commonly used as a cancer-bearing model animal, and a 6-week-old mouse was purchased and bred for 1 week, and then used for the test.
Mice were raised under a 12-hour light / dark cycle, temperature of 21 to 25 ° C., and humidity of 40 to 75%, and were allowed to freely ingest powdered food and water for breeding sterile animals.

2. Selection of tumor cells and pancreat-1 of human pancreatic cancer cells was selected to test the combined effect of the cultured PAI-1 inhibitor and the cytotoxic agent gemcitabine. In addition, HCT116 of human colon cancer cells was selected for the test of the combined effect with irinotecan. Panc-1 cells and HCT116 cells were obtained from DS Pharma Biomedical Co., Ltd.
Panc-1 cells and HCT116 cells were subcultured according to a conventional method using Dulbecco's Modified Eagle's medium containing 10% fetal bovine serum (FBS) at 37 ° C. in 95% air-5% CO 2. The cell concentration for transplantation into mice was adjusted to 1 × 10 ⁷ cells / 0.1 mL serum-free medium at the time of use.

3. Preparation of Compound and Test Substance Mixed Feed Gemcitabine used was a lyophilized preparation of gemcitabine hydrochloride (200 mg as gemcitabine in one vial) sold by Eli Lilly Japan. As irinotecan, a lyophilized preparation of irinotecan hydrochloride (100 mg of irinotecan in one vial) manufactured by Yakult Honsha Co., Ltd. was used.
PAI-1 inhibitors include [5-[[6- [4-phenyl-6- (phenylmethoxy) -2-pyrimidinyl] -2-naphthalenyl] oxy] pentyl] propanedioic acid, 2Na salt (SK-116) and [5-[[6- [5- (1,1-dimethylethyl) -2-benzoxazolyl] -2-naphthalenyl] oxy] pentyl] propanedioic acid, 2Na salt (SK-216) Company Shizuoka Caffeine Industry) was used. SK-116 and SK-216 are white dry powders. SK-116 and SK-216 were mixed with sterile animal breeding powder feed CL-2 (Claire Japan) so that the content was 50 ppm, 100 ppm and 150 ppm according to the test plan.

4). Tumor xenograft test mice were inoculated with 0.1 mL of tumor cell Kendaku solution (1 × 10 ⁷ cells) per mouse on the right flank. The tumors were allowed to grow for about 3 weeks and then divided into 10 groups.
Tumor volume was calculated using the formula (L × W ² ) / 2. Here, L represents the major axis of the tumor, and W represents the minor axis of the tumor.
During the test period, once a day, all mice were confirmed to be alive or dead, and the general condition was observed. Food intake and body weight were measured once a week. Tumor diameter was measured twice a week and the tumor volume was calculated.

5). Evaluation Method of Antitumor Effect The tumor volume over time was shown in a graph, and the effect of treatment was evaluated based on the inhibition rate of tumor growth obtained by the following formula.
Tumor growth inhibition rate (%) = 100− [100 × (T−T ₀ ) / (C−C ₀ )]
Here, T is the average tumor volume of the treatment group on the drug administration end date, and T ₀ is the average tumor volume of the same group on the drug administration start date.
C is the mean tumor volume of the control group medication end date, C ₀ is the mean tumor volume of the control group medication start date.

6). Test plan Table 2 shows the test plan of the combined effect of PAI-1 inhibitor and gemcitabine using Panc-1 pancreatic cancer cells and the combined effect of PAI-1 inhibitor and irinotecan using HCT116 colon cancer cells Table 3 shows. During the test period, the control group was allowed to freely ingest a feed containing no test substance.

Test example 1
Method Using the materials described in detail in the above “Materials and Methods”, the following tests were performed according to each method.
Each nude mouse was inoculated with 0.1 mL of tumor cell kendaku solution (1 × 10 ⁷ cells) subcutaneously on the right flank. Three weeks later, the animals were grouped as 10 animals per group. The average tumor volume for each group was 138 ± 15 mm ³ .
In this test, the gemcitabine administration group was administered intraperitoneally with 80 mg / kg of gemcitabine once every 4 days × 3 schedule. In the PAI-1 inhibitor administration group, a feed prepared by mixing SK-216 at a concentration of 100 ppm was prepared, and the SK-216 combination feed was freely consumed for 44 days from the start to the end of the test. In the combination group of PAI-1 inhibitor and gemcitabine, gemcitabine was administered intraperitoneally at 80 mg / kg once every 4 days × 3 schedule, and for 44 days from the start to the end of administration, SK-216 was 50 ppm or Each feed mixed at a concentration of 100 ppm was freely ingested. In the control group, a feed containing no test substance was ingested freely.

2. Results During the test period, once a day, all nude mice were confirmed to be alive or dead, and the general condition was observed. During the study period, there were no deaths in each group and the health status was good.
Food intake and body weight were measured once a week. As shown in FIG. 1, there was no statistically significant difference in the body weight (average weight) of each group of mice throughout the test period, and no adverse events were observed due to the combined administration of PAI-1 inhibitor and cytotoxic agent. It was.
The tumor diameter was measured twice a week, and the tumor volume of all mice was calculated from the formula of (L × W ² ) / 2. The change with time of the tumor volume (average value of each group) is shown in FIG. The inhibition rate of tumor growth was calculated based on the tumor volume of each group. The tumor growth inhibition rate was 33.5% in the gemcitabine 80 mg / kg single administration group, whereas the “SK-216 (100 ppm) + gemcitabine 80 mg / kg” combination group was about 50%, and PAI-1 inhibition It was confirmed that gemcitabine potentiated the tumor growth inhibitory effect of the combination of drugs.
From the above results, it was confirmed that the combined use of PAI-1 inhibitor and gemcitabine did not cause any adverse events, was highly safe, and the combined use of PAI-1 inhibitor enhanced the tumor growth inhibitory effect of gemcitabine It was done.

Test example 2
Method Using the materials described in detail in the above “Materials and Methods”, the following tests were performed according to each method.
Each nude mouse was inoculated with 0.1 mL of tumor cell kendaku solution (1 × 10 ⁷ cells) subcutaneously on the right flank. Three weeks later, the animals were grouped as 10 animals per group. The average tumor volume for each group was 226 ± 20 mm ³ .
In this test, the irinotecan-administered group was administered 40 mg / kg of irinotecan intravenously via the tail vein once every 4 days × 3 schedules.
The PAI-1 inhibitor administration group prepared a feed in which SK-116 was mixed at a concentration of 100 ppm, and freely ingested the SK-116 combined feed for 44 days from the start to the end of the test. In the combination group of PAI-1 inhibitor and irinotecan, irinotecan 40 mg / kg was administered into the tail vein once every 4 days × 3 schedule, and SK-116 was administered at 100 ppm or 150 ppm for 36 days from the start to the end of administration. Each of the feeds mixed at a concentration of 1 was freely ingested. In the control group, a feed containing no test substance was ingested freely.

2. Results During the test period, once a day, all nude mice were confirmed to be alive or dead, and the general condition was observed. During the study period, there were no deaths in each group and the health status was good.
Food intake and body weight were measured once a week. As shown in FIG. 3, there was no statistically significant difference in the body weight (average weight) of the mice in each group throughout the test period, and no adverse events were observed due to the combined administration of the PAI-1 inhibitor and irinotecan.
The tumor diameter was measured twice a week, and the tumor volume of all mice was calculated from the formula of (L × W ² ) / 2. The change with time of the tumor volume (average value of each group) is shown in FIG. The inhibition rate of tumor growth was calculated based on the tumor volume of each group. The tumor growth inhibition rate of the irinotecan 40 mg / kg single administration group was 51.1%, whereas the “SK-116 (100 ppm) + irinotecan 40 mg / kg” combination group was 66.3%, and PAI-1 It was confirmed that irinotecan enhanced the tumor growth-suppressing effect by the combined use of an inhibitor.
From the above results, it was confirmed that the combined use of the PAI-1 inhibitor and irinotecan did not cause any adverse events, was highly safe, and the combined use of the PAI-1 inhibitor enhanced the tumor growth inhibitory effect of irinotecan. It was done.

A combination of a PAI-1 inhibitor and a cytotoxic agent or cytostatic agent is an antitumor agent completely different from conventional combination therapies, and is used as a prophylactic, therapeutic agent for various malignant tumors, or a prophylactic agent for metastasis In particular, it is extremely useful industrially because it exhibits a high tumor growth inhibitory effect on pancreatic cancer cells when used in combination with gemcitabine. In addition, the combined effect can reduce the amount of a cytotoxic agent or cytostatic agent having strong side effects, which is beneficial from the viewpoint of the patient's “Quality of Life” (QOL).

FIG. 1 is a graph showing the weight fluctuation of nude mice during the test period in Test Example 1. FIG. 2 shows the effect of gemcitabine (Gemzar) alone and in combination with a PAI-1 inhibitor (SK-216). Nude mice were transplanted with Panc-1 pancreatic cancer tumor cells. Three weeks later, each group was divided into 10 animals so that the average tumor volume of each group was 138 ± 15 mm ³ . Each group was administered test substance and concomitant drug gemcitabine as set in the test plan. The graph shows the change over time in the tumor volume (mean value) of nude mice in each group. FIG. 3 is a graph showing the weight fluctuation of nude mice during the test period in Test Example 2. FIG. 4 shows the effect of irinotecan (CPT-11) alone and in combination with a PAI-1 inhibitor (SK-116). Nude mice were transplanted with HCT116 colon cancer tumor cells. Three weeks later, each group was divided into 10 animals so that the average tumor volume of each group was 226 ± 20 mm ³ . Each group was administered test substance and concomitant drug irinotecan as set in the test plan. The graph shows the change over time in the tumor volume (mean value) of nude mice in each group.

Claims (34)

A pharmaceutical comprising a combination of a plasminogen activator inhibitor-1 (PAI-1) inhibitor and a cytotoxic agent or cytostatic agent. A PAI-1 inhibitor is represented by the following formula (1):

[In the formula (1), ring A represents benzoxazole, benzothiazole or pyrimidine; R ₁ represents a hydroxyl group, a linear or branched alkyl group having 1 to 6 carbon atoms, and a straight chain having 1 to 4 carbon atoms. A chain or branched lower alkoxy group, a cyclohexylmethoxy group, a benzyloxy group (wherein phenyl of the benzyloxy group is a hydroxyl group, a linear or branched lower alkyl group having 1 to 4 carbon atoms, a 1- 4 linear or branched lower alkoxy groups, trifluoromethyl group, nitro group, —N (R ₄ , R ₅ ) group (where R ₄ and R ₅ are each independently hydrogen atom, carbon Represents a straight-chain or branched alkyl group or benzyl group having a number of 1-6, and may be substituted with a substituent selected from a halogen atom), a trifluoromethyl group, B group, -N _(R 4, _{R 5)} group _(wherein, R 4, _{R 5} are the same as defined above.), A halogen atom, a phenyl group (wherein the phenyl group is hydroxyl, C 1 -C -4 linear or branched lower alkyl groups, linear or branched lower alkoxy groups having 1 to 4 carbon atoms, trifluoromethyl group, nitro group, -N (R ₄ , R ₅ ) group ( Wherein R ₄ and R ₅ are the same as defined above) and may be substituted with a substituent selected from a halogen atom; and R ₂ and R ₃ are each independently hydrogen. Represents an atom or a linear or branched lower alkyl group having 1 to 4 carbon atoms; m represents 0 or an integer of 1 to 4, and m R _{1 s} may be the same or different; Represents an integer of 1 to 6. ]
The pharmaceutical according to claim 1, which is a propanedioic acid derivative represented by the formula: or a pharmaceutically acceptable salt thereof. In the PAI-1 inhibitor, ring A is benzoxazole or pyrimidine; R ₁ is a linear or branched alkyl group having _{1 to} 6 carbon atoms, a cyclohexylmethoxy group, a benzyloxy group (here, benzyloxy group) The phenyl of the oxy group may be substituted with a halogen atom) or a phenyl group (wherein the phenyl group may be substituted with a linear or branched lower alkoxy group having 1 to 4 carbon atoms). R ₂ and R ₃ are hydrogen atoms; m is an integer of 1 or 2; two R _{1 s} may be the same or different; and W is 3 to 5 The pharmaceutical according to claim 1 or 2, which is an propanedioic acid derivative represented by formula (1), or a pharmaceutically acceptable salt thereof, which is an integer. The medicament according to any one of claims 1 to 3, wherein the PAI-1 inhibitor is a compound selected from the following group, or a pharmaceutically acceptable salt thereof.
[3-[[6- (5-phenyl-2-benzoxazolyl) -2-naphthalenyl] oxy] propyl] propanedioic acid,
[5-[[6- [5- (1,1-dimethylethyl) -2-benzoxazolyl] -2-naphthalenyl] oxy] pentyl] propanedioic acid,
[5-[[6- [6- (cyclohexylmethoxy) -2-benzoxazolyl] -2-naphthalenyl] oxy] pentyl] propanedioic acid,
[5-[[6- [6- (phenylmethoxy) -2-benzoxazolyl] -2-naphthalenyl] oxy] pentyl] propanedioic acid,
[3-[[6- [4-[(2-fluorophenyl) methoxy] -6- (4-methoxyphenyl) -2-pyrimidinyl] -2-naphthalenyl] oxy] propyl] propanedioic acid,
[5-[[6- [4-Phenyl-6- (phenylmethoxy) -2-pyrimidinyl] -2-naphthalenyl] oxy] pentyl] propanedioic acid,
[5-[[6- [4-cyclohexylmethoxy-6- (1,1-dimethylethyl) -2-pyrimidinyl] -2-naphthalenyl] oxy] pentyl] propanediacid,
[5-[[6- [4- (1,1-dimethylethyl) -6-[(4-fluorophenyl) methoxy] -2-pyrimidinyl] -2-naphthalenyl] oxy] pentyl] propanedioic acid and [5 -[[6- [4-[(2-chlorophenyl) methoxy] -6- (1,1-dimethylethyl) -2-pyrimidinyl] -2-naphthalenyl] oxy] pentyl] propanedioic acid. The PAI-1 inhibitor is [5-[[6- [5- (1,1-dimethylethyl) -2-benzoxazolyl] -2-naphthalenyl] oxy] pentyl] propanedioic acid, or pharmaceutically The medicine according to any one of claims 1 to 4, which is an acceptable salt thereof. PAI-1 inhibitors are [5-[[6- [4-phenyl-6- (phenylmethoxy) -2-pyrimidinyl] -2-naphthalenyl] oxy] pentyl] propanedioic acid, or pharmaceutically acceptable The pharmaceutical according to any one of claims 1 to 4, which is a salt thereof. A cytotoxin or cytostatic agent is a DNA synthesis inhibitor, a DNA topoisomerase type I inhibitor, a DNA topoisomerase type II inhibitor, a microtubule agent, a vascular endothelial growth factor inhibitor, an epidermal growth factor receptor inhibitor, The medicine according to any one of claims 1 to 6, which is an alkylating agent, a platinum complex, an anticancer antibiotic, or a hormone therapeutic agent. Cytotoxins or cytostatics are gemcitabine, capecitabine, 5-fluorouracil, methotrexate, irinotecan, etoposide, doxorubicin, vinorelbine, paclitaxel, vincristine, vinblastine, docetaxel, bevacizumab, gefitinib trasfumab, erlotizibras The medicine according to any one of claims 1 to 7, which is cyclophosphamide, cisplatin, carboplatin, oxaliplatin, mitomycin C, actinomycin D, tamoxifen, or anastrozole. The pharmaceutical according to any one of claims 1 to 8, wherein the cytotoxic agent is gemcitabine. The pharmaceutical according to any one of claims 1 to 8, wherein the cytotoxic agent is irinotecan. The medicament according to any one of claims 1 to 10, wherein a PAI-1 inhibitor and a cytotoxic agent or a cytostatic agent are administered together. The medicament according to any one of claims 1 to 10, wherein a PAI-1 inhibitor and a cytotoxic agent or a cytostatic agent are administered separately. The medicament according to any one of claims 1 to 12, for suppressing tumor growth and / or metastasis. Tumor is lung cancer, breast cancer, pancreatic cancer, liver cancer, bile duct cancer, esophageal cancer, stomach cancer, duodenal cancer, small intestine cancer, large intestine (colon, rectum) cancer, uterine cancer, ovarian cancer, The medicament according to claim 13, which is prostate cancer, leukemia, melanoma, or malignant lymphoma. The medicament according to claim 13 or 14, wherein the tumor is pancreatic cancer or colon (colon, rectal) cancer. Use of a combination of a PAI-1 inhibitor and a cytotoxin or cytostatic agent to produce a medicament for inhibiting tumor growth and / or metastasis. The use according to claim 16, which is a medicament for administering a PAI-1 inhibitor and a cytotoxic agent or cytostatic agent together or separately. The use according to claim 16 or 17, wherein the PAI-1 inhibitor is the PAI-1 inhibitor according to any one of claims 2 to 6. The use according to any one of claims 16 to 18, wherein the cytotoxic agent or cytostatic agent is the cytotoxic agent or cytostatic agent according to any one of claims 7 to 10. A pharmaceutical kit comprising a PAI-1 inhibitor and a cytotoxic agent or a cytostatic agent. 21. The kit according to claim 20, for administering the PAI-1 inhibitor and the cytotoxic agent or cytostatic agent together or separately. The kit according to claim 20 or 21, wherein the PAI-1 inhibitor is the PAI-1 inhibitor according to any one of claims 2 to 6. The kit according to any one of claims 20 to 22, wherein the cytotoxic agent or the cell growth inhibitor is the cytotoxic agent or the cell growth inhibitor according to any one of claims 7 to 10. 24. The kit according to any one of claims 20 to 23 for suppressing tumor growth and / or metastasis. The kit according to claim 24, wherein the tumor is the tumor according to claim 14 or 15. A method for suppressing tumor growth and / or metastasis, comprising administering a PAI-1 inhibitor and a cytotoxic agent or cytostatic agent to mammals including humans. 27. The method of claim 26, wherein the PAI-1 inhibitor and the cytotoxic agent or cytostatic agent are administered together or separately. The method according to claim 26 or 27, wherein the PAI-1 inhibitor is the PAI-1 inhibitor according to any one of claims 2 to 6. The method according to any one of claims 26 to 28, wherein the cytotoxic agent or cytostatic agent is the cytotoxic agent or cytostatic agent according to any one of claims 7 to 10. The method according to any one of claims 26 to 29, wherein the tumor is the tumor according to claim 14 or 15. A pharmaceutical agent containing a PAI-1 inhibitor as an active ingredient for enhancing the tumor growth and / or metastasis-suppressing effect of a cytotoxin or cytostatic agent. The medicament according to claim 31, wherein the PAI-1 inhibitor is the PAI-1 inhibitor according to any one of claims 2 to 6. The pharmaceutical agent according to claim 31 or 32, wherein the cytotoxic agent or cytostatic agent is the cytotoxic agent or cytostatic agent according to any one of claims 7 to 10. The medicament according to any one of claims 31 to 33, wherein the tumor is the tumor according to claim 14 or 15.

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