Cancer treatment drugs

Abstract

To provide means to improve the therapeutic effect of an anticancer drug.SOLUTION: Provided is a therapeutic agent for the drug resistance of cells to microtubule inhibitors, the therapeutic agent containing a κ opioid receptor agonist.SELECTED DRAWING: Figure 12

Description

[Technical Field]

[0001] This invention relates to a therapeutic agent for treating cellular drug resistance to microtubule inhibitors, and to a pharmaceutical composition for cancer treatment. [Background technology]

[0002] Taxane-based anticancer drugs are used in cancer chemotherapy. Examples of taxane-based anticancer drugs include paclitaxel and docetaxel. When paclitaxel and docetaxel are administered to cancer patients, drug resistance tends to develop in the patient's cancer cells, leading to treatment resistance and a decrease in treatment efficacy. Another taxane-based anticancer drug is cabazitaxel. For example, cabazitaxel is effective even in prostate cancer patients with a history of prior treatment including docetaxel. However, drug resistance also tends to develop in cancer cells to cabazitaxel, leading to treatment resistance and a decrease in treatment efficacy (see, for example, Patent Document 1). [Prior art documents] [Patent Documents]

[0003] [Patent Document 1] Patent No. 6735484 [Overview of the Initiative] [Problems that the invention aims to solve]

[0004] There is a need for means to enhance the therapeutic effect of anticancer drugs against all types of cancer, not just prostate cancer. Therefore, one of the objectives of this invention is to provide means to enhance the therapeutic effect of anticancer drugs. [Means for solving the problem]

[0005] According to aspects of the present invention, a therapeutic agent for treating cellular drug resistance to microtubule inhibitors is provided, comprising a κ-opioid receptor agonist.

[0006] The above-mentioned therapeutic agent for treating cellular drug resistance to microtubule inhibitors may contain a κ-opioid receptor agonist as an active ingredient.

[0007] The above-mentioned therapeutic agent for treating cellular drug resistance to microtubule inhibitors may contain an effective amount of a κ-opioid receptor agonist for treating cellular drug resistance to microtubule inhibitors.

[0008] In the therapeutic agent for treating cellular drug resistance to the above-mentioned microtubule inhibitors, the κ-opioid receptor agonist may be at least one selected from the group consisting of spiradrin, nalfurafine, ICI-199,441, LPK-26, BRL-52537, and GR-89696, as well as salts thereof.

[0009] In the therapeutic agent for treating cellular drug resistance to the above-mentioned microtubule inhibitors, the κ-opioid receptor agonist may be at least one of spiradrin and its salts.

[0010] In the therapeutic agent for treating cellular drug resistance to the above-mentioned microtubule inhibitors, the salt may be a physiologically acceptable salt.

[0011] In the above-mentioned therapeutic agent for treating cellular drug resistance to microtubule inhibitors, the microtubule inhibitor may be a taxane-based anticancer agent.

[0012] In the above-mentioned therapeutic agent for treating cellular drug resistance to microtubule inhibitors, the taxane-based anticancer agent may be at least one selected from the group consisting of cabazitaxel, paclitaxel, and docetaxel.

[0013] In the therapeutic agent for treating cellular drug resistance to the above-mentioned microtubule inhibitors, the taxane-based anticancer agent may be cabazitaxel.

[0014] In the therapeutic agent for treating cellular drug resistance to the above-mentioned microtubule inhibitors, the cancer may be at least one selected from the group consisting of prostate cancer, bladder cancer, breast cancer, gastric cancer, and uterine cancer.

[0015] In the therapeutic agent for drug resistance of cells to the above-mentioned microtubule inhibitor, the cancer may be a urinary tract cancer. The urinary tract cancer may be at least one selected from prostate cancer, bladder cancer, kidney cancer, ureteropelvic cancer, testicular cancer, and penile cancer.

[0016] In the therapeutic agent for drug resistance of cells to the above-mentioned microtubule inhibitor, the cancer may be prostate cancer. The cancer may be breast cancer. The cancer may be uterine cancer.

[0017] The above-mentioned therapeutic agent for drug resistance of cells to the microtubule inhibitor may be administered to a cancer patient. The patient may be a human or a non-human animal.

[0018] Also, according to an aspect of the present invention, there is provided the use of a κ-opioid receptor agonist in the production of a therapeutic agent for drug resistance of cells to a microtubule inhibitor.

[0019] In the above-mentioned use, the therapeutic agent for drug resistance of cells to the microtubule inhibitor may contain a κ-opioid receptor agonist as an active ingredient.

[0020] In the above-mentioned use, the therapeutic agent for drug resistance of cells to the microtubule inhibitor may contain an effective amount of a κ-opioid receptor agonist for treating drug resistance of cells to the microtubule inhibitor.

[0021] In the above-mentioned use, the κ-opioid receptor agonist may be at least one selected from the group consisting of spiradoline, nalorphine, ICI-199,441, LPK-26, BRL-52537, and GR-89696, and salts thereof.

[0022] In the above-mentioned use, the κ-opioid receptor agonist may be at least any one of spiradoline and its salts.

[0023] In the above use, the salt may be a physiologically acceptable salt.

[0024] In the above use, the microtubule inhibitor may be a taxane-based anticancer agent.

[0025] In the above use, the taxane-based anticancer agent may be at least one selected from the group consisting of cabazitaxel, paclitaxel, and docetaxel.

[0026] In the above use, the taxane-based anticancer agent may be cabazitaxel.

[0027] In the above use, cancer may be at least one selected from the group consisting of prostate cancer, bladder cancer, breast cancer, stomach cancer, and uterine cancer.

[0028] In the above use, cancer may be a urinary tract cancer. Urinary tract cancer may be at least one selected from prostate cancer, bladder cancer, kidney cancer, renal pelvis and ureteral cancer, testicular cancer, and penile cancer.

[0029] In the above use, cancer may be prostate cancer. Cancer may be breast cancer. Cancer may be uterine cancer.

[0030] In the above use, a therapeutic agent for treating cellular drug resistance to microtubule inhibitors may be administered to a cancer patient. The patient may be a human or a non-human animal.

[0031] Furthermore, according to an aspect of the present invention, a method is provided for treating cellular drug resistance to microtubule inhibitors, comprising administering a κ-opioid receptor agonist to a patient.

[0032] In the above-mentioned method for treating cellular drug resistance to microtubule inhibitors, the patient may be administered a κ-opioid receptor agonist as the active ingredient.

[0033] In the above-described method for treating cellular drug resistance to microtubule inhibitors, the patient may be administered an effective dose of a κ-opioid receptor agonist to treat the cellular drug resistance to the microtubule inhibitor.

[0034] In the above-mentioned method for treating cellular drug resistance to microtubule inhibitors, the κ-opioid receptor agonist may be at least one selected from the group consisting of spiradrin, nalfurafine, ICI-199,441, LPK-26, BRL-52537, and GR-89696, as well as salts thereof.

[0035] In the method for treating cellular drug resistance to the above-mentioned microtubule inhibitors, the κ-opioid receptor agonist may be at least one of spiradrin and its salts.

[0036] In the above-mentioned method for treating cellular drug resistance to microtubule inhibitors, the salt may be a physiologically acceptable salt.

[0037] In the above-mentioned method for treating cellular drug resistance to microtubule inhibitors, the microtubule inhibitor may be a taxane-based anticancer drug.

[0038] In the above-mentioned method for treating drug resistance of cells to microtubule inhibitors, the taxane-based anticancer agent may be at least one selected from the group consisting of cabazitaxel, paclitaxel, and docetaxel.

[0039] In the above-mentioned method for treating drug resistance of cells to microtubule inhibitors, the taxane-based anticancer agent may be cabazitaxel.

[0040] In the above-mentioned method for treating drug resistance of cells to microtubule inhibitors, the cancer may be at least one selected from the group consisting of prostate cancer, bladder cancer, breast cancer, gastric cancer, and uterine cancer.

[0041] In the above-described method for treating drug resistance of cells to microtubule inhibitors, the cancer may be a urinary tract cancer. The urinary tract cancer may be at least one selected from prostate cancer, bladder cancer, kidney cancer, renal pelvis and ureteral cancer, testicular cancer, and penile cancer.

[0042] In the above-mentioned method for treating drug resistance of cells to microtubule inhibitors, the cancer may be prostate cancer, breast cancer, or uterine cancer.

[0043] In the above-mentioned method for treating drug resistance of cells to microtubule inhibitors, the patient may be a cancer patient. The patient may be a human or a non-human animal.

[0044] Furthermore, according to an aspect of the present invention, a pharmaceutical composition for cancer treatment is provided, comprising a κ-opioid receptor agonist and a microtubule inhibitor.

[0045] The above-mentioned cancer treatment drug composition may contain a κ-opioid receptor agonist as an active ingredient for treating cellular drug resistance to microtubule inhibitors.

[0046] The above-described cancer treatment drug composition may contain an effective amount of a κ-opioid receptor agonist for treating cellular drug resistance to microtubule inhibitors.

[0047] In the above-described cancer treatment drug composition, the κ-opioid receptor agonist may be at least one selected from the group consisting of spiradrin, nalfurafine, ICI-199,441, LPK-26, BRL-52537, and GR-89696, and salts thereof.

[0048] In the above-described cancer treatment drug composition, the κ-opioid receptor agonist may be at least one of spiradrine and its salts.

[0049] In the above-described cancer treatment drug composition, the salt may be a physiologically acceptable salt.

[0050] In the above-described cancer treatment drug composition, the microtubule inhibitor may be a taxane-based anticancer agent.

[0051] In the above-described cancer treatment drug composition, the taxane-based anticancer agent may be at least one selected from the group consisting of cabazitaxel, paclitaxel, and docetaxel.

[0052] In the above-described cancer treatment drug composition, the taxane-based anticancer agent may be cabazitaxel.

[0053] In the above-described cancer treatment drug composition, the cancer may be at least one selected from the group consisting of prostate cancer, bladder cancer, breast cancer, stomach cancer, and uterine cancer.

[0054] In the above-described drug composition for cancer treatment, the cancer may be a urinary tract cancer. The urinary tract cancer may be at least one selected from prostate cancer, bladder cancer, kidney cancer, renal pelvis and ureteral cancer, testicular cancer, and penile cancer.

[0055] In the above-described cancer treatment drug composition, the cancer may be prostate cancer. The cancer may be breast cancer. The cancer may be uterine cancer.

[0056] Furthermore, according to aspects of the present invention, the use of a κ-opioid receptor agonist and a microtubule inhibitor in the manufacture of a pharmaceutical composition for cancer treatment is provided.

[0057] In the above use, the cancer treatment drug composition may contain a κ-opioid receptor agonist as an active ingredient for treating cellular drug resistance to microtubule inhibitors.

[0058] In the above use, the cancer treatment drug composition may contain an effective amount of a κ-opioid receptor agonist for treating cellular drug resistance to microtubule inhibitors.

[0059] In the above use, the κ-opioid receptor agonist may be at least one selected from the group consisting of spiradrin, nalfurafine, ICI-199,441, LPK-26, BRL-52537, and GR-89696, and their salts.

[0060] In the above use, the κ-opioid receptor agonist may be at least one of spiradrine and its salts.

[0061] In the above use, the salt may be a physiologically acceptable salt.

[0062] In the above use, the microtubule inhibitor may be a taxane-based anticancer agent.

[0063] In the above use, the taxane-based anticancer agent may be at least one selected from the group consisting of cabazitaxel, paclitaxel, and docetaxel.

[0064] In the above use, the taxane-based anticancer agent may be cabazitaxel.

[0065] In the above use, cancer may be at least one selected from the group consisting of prostate cancer, bladder cancer, breast cancer, stomach cancer, and uterine cancer.

[0066] In the above use, cancer may be a urinary tract cancer. Urinary tract cancer may be at least one selected from prostate cancer, bladder cancer, kidney cancer, renal pelvis and ureteral cancer, testicular cancer, and penile cancer.

[0067] In the above use, cancer may be prostate cancer. Cancer may be breast cancer. Cancer may be uterine cancer.

[0068] Furthermore, according to an aspect of the present invention, a method for treating cancer is provided, comprising administering a κ-opioid receptor agonist and a microtubule inhibitor to a patient.

[0069] In the above-described cancer treatment method, patients may be administered a κ-opioid receptor agonist as an active ingredient to treat cellular drug resistance to microtubule inhibitors.

[0070] In the above-described cancer treatment method, the patient may be administered an effective dose of a κ-opioid receptor agonist to treat the cellular drug resistance to microtubule inhibitors.

[0071] In the above-described method for treating cancer, the κ-opioid receptor agonist may be at least one selected from the group consisting of spiradrin, nalfurafine, ICI-199,441, LPK-26, BRL-52537, and GR-89696, as well as salts thereof.

[0072] In the above-described method for treating cancer, the κ-opioid receptor agonist may be at least one of spiradrin and its salts.

[0073] In the cancer treatment methods described above, the salt may be physiologically acceptable salt.

[0074] In the above-mentioned cancer treatment method, the microtubule inhibitor may be a taxane-based anticancer drug.

[0075] In the above-described method of treating cancer, the taxane-based anticancer drug may be at least one selected from the group consisting of cabazitaxel, paclitaxel, and docetaxel.

[0076] In the above-mentioned cancer treatment method, the taxane-based anticancer drug may be cabazitaxel.

[0077] In the above-described method of treating cancer, the cancer may be at least one selected from the group consisting of prostate cancer, bladder cancer, breast cancer, stomach cancer, and uterine cancer.

[0078] In the above-described cancer treatment method, the cancer may be a urinary tract cancer. The urinary tract cancer may be at least one selected from prostate cancer, bladder cancer, kidney cancer, renal pelvis and ureteral cancer, testicular cancer, and penile cancer.

[0079] In the cancer treatment methods described above, the cancer may be prostate cancer, breast cancer, or uterine cancer.

[0080] In the cancer treatment methods described above, the patient may be a human or a non-human animal. [Effects of the Invention]

[0081] According to the present invention, it is possible to provide a means to enhance the therapeutic effect of anticancer drugs. [Brief explanation of the drawing]

[0082] [Figure 1] Figure 1 is a table showing the combination compounds used in Example 1. [Figure 2] Figure 2 is a graph showing the relationship between the concentration of cabazitaxel and the survival rate of DU145 cells that are not resistant to taxane-based anticancer drugs, in Example 1. [Figure 3] Figure 3 is a graph showing the relationship between the concentration of cabazitaxel and the survival rate of docetaxel-resistant DU145 cells in Example 1. [Figure 4] Figure 4 is a graph showing the relationship between the concentration of cabazitaxel and the survival rate of cabazitaxel-resistant DU145 cells in Example 1. [Figure 5] Figure 5 is a table showing the IC50 of cabazitaxel according to Example 1. [Figure 6] Figure 6 is a graph showing the relationship between the concentration of cabazitaxel and the survival rate of docetaxel-resistant PC3 cells in Example 2. [Figure 7] Figure 7 is a graph showing the relationship between the concentration of cabazitaxel and the survival rate of cabazitaxel-resistant PC3 cells in Example 2. [Figure 8] Figure 8 is a table showing the IC50 of cabazitaxel in Example 2. [Figure 9] Figure 9 is a graph showing the relationship between the concentration of paclitaxel and the survival rate of docetaxel-resistant PC3 cells in Example 3. [Figure 10] Figure 10 is a graph showing the relationship between the concentration of docetaxel and the survival rate of docetaxel-resistant PC3 cells in Example 3. [Figure 11] Figure 11 is a table showing the IC50 of cabazitaxel when used in combination with a κ-opioid receptor agonist, according to Example 4. [Figure 12] Figure 12 is a graph showing the change in tumor size over time in mice according to Example 5. [Figure 13] Figure 13 is a graph showing the relationship between the concentration of paclitaxel and the survival rate of paclitaxel-resistant MCF7-TxR cells in Example 7. [Figure 14] Figure 14 is a graph showing the relationship between the concentration of docetaxel and the survival rate of paclitaxel-resistant MCF7-TxR cells in Example 7. [Figure 15] Figure 15 is a graph showing the relationship between the concentration of paclitaxel and the survival rate of multidrug-resistant MES-SA / Dx5 cells in Example 7. [Figure 16] Figure 16 is a graph showing the relationship between the concentration of docetaxel and the survival rate of multidrug-resistant MES-SA / Dx5 cells in Example 7. [Modes for carrying out the invention]

[0083] A therapeutic agent for treating cellular drug resistance to microtubule inhibitors according to an embodiment of the present invention includes a κ-opioid receptor agonist.

[0084] Microtubule inhibitors inhibit cell division by inhibiting the depolymerization of microtubules in cells. Microtubules play a crucial role in cell division. In cells, microtubules polymerize with each other until metaphase. In anaphase, microtubules depolymerize within the cell. Microtubule inhibitors inhibit cell division by inhibiting the depolymerization of microtubules in cells.

[0085] Cancer cells proliferate by repeatedly dividing. Microtubule inhibitors inhibit the cell division of cancer cells and can therefore be used as anticancer agents. Microtubule inhibitors are, for example, taxane-based anticancer agents. Taxane-based anticancer agents have a taxane ring. Examples of taxane-based anticancer agents include cabazitaxel, paclitaxel, and docetaxel. Cancers targeted by taxane-based anticancer agents include, for example, urinary tract cancers. Examples of urinary tract cancers include prostate cancer, bladder cancer, kidney cancer, renal pelvis and ureteral cancer, testicular cancer, and penile cancer. Cancers targeted by taxane-based anticancer agents are not limited to urinary tract cancers, but may also include breast cancer, uterine cancer, stomach cancer, ovarian cancer, non-small cell lung cancer, gastric cancer, and esophageal cancer, etc.

[0086] When cancer patients are administered microtubule inhibitors, cancer cells tend to develop drug resistance to the inhibitors, leading to treatment resistance and a decrease in the therapeutic effect of the microtubule inhibitors. In contrast, κ-opioid receptor agonists treat the patient's resistance to microtubule inhibitors.

[0087] Examples of κ-opioid receptor agonists include spiradrine, nalfurafine, ICI-199,441, LPK-26, BRL-52537, and GR-89696. κ-opioid receptor agonists may also be physiologically acceptable salts of these.

[0088] The IUPAC name for spiradrin (CAS number: 87151-85-7) is 2-(3,4-dichlorophenyl)-N-methyl-N-[(5R,7S,8S)-7-(pyrrolidin-1-yl)-1-oxaspiro[4.5]decan-8-yl]acetamide. The chemical formula for spiradrin is C 22 H 30 It is Cl2N2O2. The chemical structure of spiradrin is as follows: [ka]

[0089] The IUPAC name for nalfurafine (CAS number: 152658-17-8) is (E)-N-[(4R,4aS,7R,7aR,12bS)-3-(cyclopropylmethyl)-4a,9-dihydroxy-1,2,4,5,6,7,7a,13-octahydro-4,12-methanobenzofuro[3,2-e]isoquinolin-7-yl]-3-(furan-3-yl)-N-methylprop-2-enamide. The chemical formula for nalfurafine is C 28 H 32 It is N2O5. The chemical structure of nalfurafin is as follows: [ka]

[0090] The IUPAC name for ICI-199,441 (CAS number: 115199-84-3) is 2-(3,4-Dichlorophenyl)-N-methyl-N-[(1S)-1-phenyl-2-(1-pyrrolidinyl)ethyl]acetamide. The chemical formula for ICI-199,441 is C 21 H 24 It is Cl2N2O. The chemical structure of ICI-199,441 is as follows: [ka]

[0091] The IUPAC name of LPK-26 (CAS No.: 492451-07-7) is 2-(3,4-dichlorophenyl)-N-[(2S)-1-(2,5-dihydropyrrol-1-yl)-3-methylbutan-2-yl]-N-methylacetamide. The chemical formula of LPK-26 is C 18 H 24 Cl2N2O. The chemical structural formula of LPK-26 is as follows.

Chem.

[0092] The IUPAC name of BRL-52537 (CAS No.: 130497-33-5) is 2-(3,4-dichlorophenyl)-1-[(2S)-2-(pyrrolidin-1-ylmethyl)piperidin-1-yl]ethanone. The chemical formula of BRL-52537 is C 18 H 24 Cl2N2O. The chemical structural formula of BRL-52537 is as follows.

Chem.

[0093] The IUPAC name of GR-89696 (CAS No.: 126766-32-3) is methyl 4-[2-(3,4-dichlorophenyl)acetyl]-3-(pyrrolidin-1-ylmethyl)piperazine-1-carboxylate. The chemical formula of GR-89696 is C 19 H 25 Cl2N3O3. The chemical structural formula of GR-89696 is as follows.

Chem.

[0094] The therapeutic agents for cellular drug resistance to microtubule inhibitors according to embodiments of the present invention can be formulated into pharmaceutically acceptable dosage forms. For example, the therapeutic agents for cellular drug resistance to microtubule inhibitors can be formulated into injections, solutions, suspensions, tablets, capsules, pills, granules, syrups, suppositories, inhalants, and sprays. Injectable agents include, for example, injection solutions, sterile powders for injection, and concentrates for injection.

[0095] Furthermore, the cancer treatment pharmaceutical composition according to the embodiment of the present invention comprises a κ-opioid receptor agonist and a microtubule inhibitor. The κ-opioid receptor agonist and the microtubule inhibitor are as described above. The κ-opioid receptor agonist and the microtubule inhibitor may be administered to cancer patients simultaneously or separately. The κ-opioid receptor agonist and the microtubule inhibitor may be administered to cancer patients at different times and on different days. The κ-opioid receptor agonist and the microtubule inhibitor may be administered to cancer patients the same number of times or at different numbers of times.

[0096] The cancer treatment pharmaceutical composition according to the embodiment of the present invention can be formulated into pharmaceutically acceptable dosage forms. For example, the cancer treatment pharmaceutical composition can be formulated into injections, solutions, suspensions, tablets, capsules, pills, granules, syrups, suppositories, inhalants, and sprays. Injections include, for example, injection solutions, sterile powders for injection, and concentrated solutions for injection. In the cancer treatment pharmaceutical composition according to the embodiment of the present invention, the κ-opioid receptor agonist and the microtubule inhibitor may be formulated separately or together.

[0097] Although the present invention has been described above with reference to embodiments, the descriptions and drawings that constitute part of this disclosure should not be understood as limiting the invention. Various alternative embodiments, examples, and operational techniques should become apparent to those skilled in the art from this disclosure. It should be understood that the present invention encompasses various embodiments and the like that are not described herein.

[0098] (Example 1) We prepared DU145 cells (ATCC), an androgen-independent human prostate cancer cell line lacking androgen receptors, as well as DU145_TxR cells resistant to paclitaxel and docetaxel, and DU145_CxR cells resistant to docetaxel and cabazitaxel. The DU145_TxR and DU145_CxR cells were established by referring to the description in Prostate, 2007, Vol. 67 (No. 9), pp. 955-967.

[0099] Place 1 × 10 cells in each well of a 96-well plate. 4 Cells were seeded at a cell-per-well ratio. DMEM medium supplemented with 10% FBS was used as the culture medium. 24 hours after seeding, the medium was replaced with one containing cabazitaxel at concentrations of 0 nmol / L, 1 nmol / L, 3 nmol / L, 10 nmol / L, 30 nmol / L, or 100 nmol / L, along with one of the combination compounds shown in Figure 1. The concentration of the combination compound was set to a level that resulted in a cell viability of 90% or more when the combination compound was added to cells alone.

[0100] Forty-eight hours after changing the culture medium, a colorimetric quantitative analysis reagent (CellTiter 96 AQueous One Solution Cell Proliferation Assay kit, Promega, G3581) for measuring the number of viable cells was added directly to the culture medium and reacted for one hour. After that, the absorbance at 490 nm was measured. The cell viability under each condition was calculated, with the cell viability under the condition of adding 0 nmol / L cabazitaxel and DMSO set as 100%. The results are shown in Figures 2, 3, and 4.

[0101] As shown in Figure 2, the effect of cabazitaxel on DU145 cells that were not resistant to taxane-based anticancer drugs was the same regardless of the combination compound used. In contrast, as shown in Figures 3 and 4, the effect of cabazitaxel in combination with spiradrin (U-62066) was superior to that of cabazitaxel in combination with other combination compounds on paclitaxel- and docetaxel-resistant DU145_TxR cells, and on DU145_CxR cells that were resistant to both docetaxel and cabazitaxel. The IC of cabazitaxel calculated from the graphs in Figures 2, 3, and 4. 50 Figure 5 shows the results. Compared to DMSO, when used in combination with spiradrin, cabazitaxel IC 50 It decreased significantly.

[0102] (Example 2) PC3 cells (ATCC), an androgen-independent human prostate cancer cell line lacking androgen receptors, paclitaxel and docetaxel-resistant PC3_TxR cells, and docetaxel and cabazitaxel-resistant PC3_CxR cells were prepared. DMSO and spiradrin in concentrations of 0.1 μmol / L, 1 μmol / L, or 10 μmol / L were prepared as concomitant compounds. Cabazitaxel was administered to these cells under the same conditions as in Example 1, except for the use of these cells and concomitant compounds, and the cell viability under each condition was determined. The results are shown in Figures 6 and 7.

[0103] As shown in Figures 6 and 7, the effect of cabazitaxel increased in a spiradlin-dependent manner against paclitaxel and docetaxel-resistant PC3_TxR cells and docetaxel and cabazitaxel-resistant PC3_CxR cells. The IC of cabazitaxel calculated from the graphs in Figures 6 and 7 50 Figure 8 shows the results. Compared to DMSO, when used in combination with spiradrin, cabazitaxel IC 50 It decreased significantly.

[0104] (Example 3) Paclitaxel and docetaxel-resistant PC3_TxR cells were prepared. DMSO and either 3 μmol / L or 10 μmol / L of spiradrin were used as co-administered compounds. Except for the use of these cells and co-administered compounds, the cells were administered paclitaxel or docetaxel under the same conditions as in Example 1, and the cell viability under each condition was determined. The results are shown in Figures 9 and 10.

[0105] As shown in Figures 9 and 10, the effects of paclitaxel and docetaxel were increased in a concentration-dependent manner against paclitaxel and docetaxel-resistant PC3_TxR cells.

[0106] (Example 4) Cabazitaxel-resistant DU145_CxR cells and PC3_CxR cells were prepared. DMSO, along with 10 μmol / L spiradrin, nalfurafine, ICI-199,441, LPK-26, BRL-52537, and GR-89696, were prepared as concomitant compounds. Cabazitaxel was administered to the cells under the same conditions as in Examples 1 and 2, except for the use of these concomitant compounds. The IC50 of cabazitaxel was determined from the cell viability under each condition and is shown in Figure 11.

[0107] As shown in Figure 11, the combined use of spiradrin, nalfurafine, ICI-199,441, LPK-26, BRL-52537, and GR-89696, respectively, enhanced the efficacy of cabazitaxel against cabazitaxel-resistant DU145_CxR cells and PC3_CxR cells.

[0108] (Example 5) Forty five-week-old male mice with severe combined immunodeficiency (SCID) were purchased from CREA Japan (Tokyo, Japan). After an acclimatization period, 3 x 10 6 PC3_CxR cells were mixed with 50% Matrigel (Corning, New York, USA) and transplanted dorsally subcutaneously into SCID mice. After the mouse tumors grew large enough to be measured in length, 36 mice were divided into six groups, excluding four mice with insufficient tumor development, so that the average tumor size in each group was approximately equal.

[0109] The first group was designated as the control group, the second group as receiving only 5 mg / kg of cabazitaxel, the third group as receiving only 2 mg / kg of spiradrine, the fourth group as receiving a combination of 5 mg / kg of cabazitaxel and 2 mg / kg of spiradrine, the fifth group as receiving only 5 mg / kg of spiradrine, and the sixth group as receiving a combination of 5 mg / kg of cabazitaxel and 5 mg / kg of spiradrine.

[0110] In groups 2, 4, and 6, which received cabazitaxel, cabazitaxel dissolved in 20 μL of DMSO was administered intraperitoneally once a week.

[0111] In groups 1, 3, and 5, which did not receive cabazitaxel, 20 μL of DMSO was administered intraperitoneally at the same frequency as in groups 2, 4, and 6, which received cabazitaxel.

[0112] In groups 3, 4, 5, and 6, who received spiradrine, spiradrine was dissolved in 20 μL of distilled water and administered intraperitoneally twice a week.

[0113] As shown in Figure 12, tumor size and mouse body weight were measured simultaneously over time using calipers and a scale, respectively. Tumor size V(cm 3 The tumor's longest diameter was A (cm) and its shortest diameter was I (cm), and the following formula was used to calculate it. V = A × I × I × 0.5 Mice were administered drugs based on their body weight over a three-week period. Afterward, the mouse with the smallest tumor size was removed from each group, and the average tumor size of five mice in each group was calculated, as shown in Figure 12. This protocol was approved by the Animal Experiment Committee of the Graduate School of Medicine, Kanazawa University.

[0114] As shown in Figure 12, the tumor size in mice administered with both spiradrine and cabazitaxel was smaller than that in mice administered with either spiradrine or cabazitaxel alone. Furthermore, the tumor size in mice administered with 5 mg / kg of spiradrine and cabazitaxel was smaller than that in mice administered with 2 mg / kg of spiradrine and cabazitaxel.

[0115] (Example 6) Using the breast cancer cell line MCF-7 as the parent cell line, we initiated the establishment of a paclitaxel-resistant cell line. DMEM medium containing 10% FCS was used as the culture medium. Paclitaxel was added to the medium at a concentration of 1 nmol / L, and the medium was changed every 2-3 days until the cells reached confluence. Once the cells reached confluence, they were subcultured, and the paclitaxel concentration was increased to 3 nmol / L, continuing the cell culture. Subsequently, the paclitaxel concentration was gradually increased, and a portion of the cells were cryopreserved after each subculture. Ultimately, a cell line capable of proliferating even at 12 nmol / L of paclitaxel was established. If cell proliferation ceased at higher concentrations, the cryopreserved cells were used to culture and subculture them with a paclitaxel concentration that ensured proliferation. The established cell line was named MCF7-TxR cells.

[0116] (Example 7) We prepared MCF7-TxR cells established in Example 6, MES-SA / Dx5 cells (a multidrug-resistant uterine cancer cell line containing paclitaxel and docetaxel), MCF-7 cells (the parent cell line of MCF7-TxR cells), and MES-SA cells (the parent cell line of MES-SA / Dx5 cells). For MES-SA / Dx5 cells, Harker WG, Sikic BI. Multidrug (pleiotropic) resistance in doxorubicin-selected variants of the human sarcoma cell line MES-SA. Cancer Res. (1985) 45:4091-4096 and Chen G, Duran GE, Steger KA, Lacayo NJ, Jaffrezou JP, Dumontet C, Sikic. See BI. Multidrug-resistant human sarcoma cells with a mutant P-glycoprotein, altered phenotype, and resistance to cyclosporins. J Biol Chem. (1997) 272:5974-5982.

[0117] 4 x 10 in a 384-well plate 3 Cells were seeded at a cell / well concentration. DMEM medium supplemented with 10% FBS was used for MCF-7 cells and MCF7-TxR cells. RPMI1640 medium supplemented with 10% FBS was used for MES-SA cells and MCF7-TxR cells. Four hours after seeding, the culture medium was changed with a medium containing either 0 nmol / L, 1 nmol / L, 3 nmol / L, 10 nmol / L, 30 nmol / L, or 100 nmol / L of paclitaxel or docetaxel, along with DMSO or 3 μmol / L of spiradrin.

[0118] Forty-eight hours after changing the culture medium, a colorimetric quantitative analysis reagent (CellTiter 96 AQueous One Solution Cell Proliferation Assay kit, Promega, G3581) for measuring the number of viable cells was added directly to the culture medium and reacted for one hour. After that, the absorbance at 490 nm was measured. The cell viability under each condition was calculated, with the cell viability under the condition of adding 0 nmol / L cabazitaxel and DMSO set as 100%.

[0119] As shown in Figure 13, when paclitaxel was added to the culture medium of non-drug-resistant MCF-7 cells, the effect of paclitaxel was almost the same whether DMSO was used or spiradrine was used. When paclitaxel was added to the culture medium of paclitaxel-resistant MCF7-TxR cells, the effect of paclitaxel was higher when spiradrine was used compared to when DMSO was used.

[0120] As shown in Figure 14, when docetaxel was added to the culture medium of non-drug-resistant MCF-7 cells, the effect of docetaxel was almost the same whether it was used in combination with DMSO or with spiradrine. When docetaxel was added to the culture medium of paclitaxel-resistant MCF7-TxR cells, the effect of docetaxel was higher when used in combination with spiradrine compared to when it was used in combination with DMSO. These results indicate that cells maintain drug resistance even when the microtubule inhibitor is changed, and that κ-opioid receptor agonists reduce the resistance of cells resistant to multiple microtubule inhibitors.

[0121] As shown in Figure 15, when paclitaxel was added to the culture medium of non-drug-resistant MES-SA cells, the effect of paclitaxel was almost the same whether it was used in combination with DMSO or with spiradrine. When paclitaxel was added to the culture medium of multidrug-resistant MES-SA / Dx5 cells, the effect of paclitaxel was higher when used in combination with spiradrine compared to when it was used in combination with DMSO.

[0122] As shown in Figure 16, when docetaxel was added to the culture medium of non-drug-resistant MES-SA cells, the effect of docetaxel was almost the same whether it was used in combination with DMSO or with spiradrine. When docetaxel was added to the culture medium of multidrug-resistant MES-SA / Dx5 cells, the effect of docetaxel was higher when used in combination with spiradrine compared to when it was used in combination with DMSO.

Claims

1. A therapeutic agent for treating cellular drug resistance to taxane-based anticancer drugs, which includes a κ-opioid receptor agonist, The κ-opioid receptor agonist is at least one selected from the group consisting of spiradrin, nalfurafine, ICI-199,441, LPK-26, BRL-52537, and GR-89696, and salts thereof. A treatment agent for treating cell resistance to taxane-based anticancer drugs.

2. A therapeutic agent for treating cellular drug resistance to a taxane-based anticancer agent according to claim 1, wherein the κ-opioid receptor agonist is at least one of spiradrin and its salts.

3. A therapeutic agent for treating cellular drug resistance to a taxane-based anticancer agent according to claim 1 or 2, wherein the taxane-based anticancer agent is at least one selected from the group consisting of cabazitaxel, paclitaxel, and docetaxel.

4. A therapeutic agent for treating cellular drug resistance to a taxane-based anticancer agent according to claim 1 or 2, wherein the taxane-based anticancer agent is cabazitaxel.

5. A therapeutic agent for treating cellular drug resistance to a taxane-based anticancer agent according to any one of claims 1 to 4, wherein the cancer is at least one selected from the group consisting of prostate cancer, bladder cancer, breast cancer, gastric cancer, and uterine cancer.

6. A therapeutic agent for treating cellular drug resistance to a taxane-based anticancer agent according to any one of claims 1 to 4, wherein the cancer is prostate cancer.

7. A therapeutic agent for treating cellular drug resistance to a taxane-based anticancer agent according to any one of claims 1 to 4, wherein the cancer is breast cancer.

8. A therapeutic agent for treating cellular drug resistance to a taxane-based anticancer agent according to any one of claims 1 to 4, wherein the cancer is uterine cancer.

9. A cancer treatment drug composition comprising a κ-opioid receptor agonist and a taxane-based anticancer agent, The κ-opioid receptor agonist is at least one selected from the group consisting of spiradrin, nalfurafine, ICI-199,441, LPK-26, BRL-52537, and GR-89696, and salts thereof. A pharmaceutical composition for the treatment of cancer in patients who are resistant to taxane-based anticancer drugs.

10. A pharmaceutical composition for treating cancer in patients resistant to taxane-based anticancer agents according to claim 9, wherein the κ-opioid receptor agonist is at least one of spiradrin and its salts.

11. A pharmaceutical composition for treating cancer in patients resistant to taxane-based anticancer agents according to claim 9 or 10, wherein the taxane-based anticancer agent is at least one selected from the group consisting of cabazitaxel, paclitaxel, and docetaxel.

12. A pharmaceutical composition for treating cancer in patients resistant to the taxane-based anticancer agent according to claim 9 or 10, wherein the taxane-based anticancer agent is cabazitaxel.

13. A pharmaceutical composition for treating cancer in patients resistant to taxane-based anticancer agents according to any one of claims 9 to 12, wherein the cancer is at least one selected from the group consisting of prostate cancer, bladder cancer, breast cancer, gastric cancer, and uterine cancer.

14. A pharmaceutical composition for treating cancer in patients who are resistant to the taxane-based anticancer agent according to any one of claims 9 to 12, wherein the cancer is prostate cancer.

15. A pharmaceutical composition for treating cancer in patients who are resistant to the taxane-based anticancer agent according to any one of claims 9 to 12, wherein the cancer is breast cancer.

16. A pharmaceutical composition for treating cancer in patients who are resistant to the taxane-based anticancer agent according to any one of claims 9 to 12, wherein the cancer is uterine cancer.

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