Patent Application
20100249089
The present invention relates to a breast cancer resistance protein (BCRP/ABCG2) inhibitor.
Serious problems associated with cancer chemotherapy include intrinsic resistance to an anticancer agent, which invalidates the effect of the anticancer agent from the beginning of cancer therapy, and development of acquired resistance to an anticancer agent (i.e., reduction of the effect of the drug, which is caused by long-term continuous administration thereof). Overcoming such resistance to anticancer agents has been envisaged to lead to improvement in the performance of cancer chemotherapy, and thus attempts have been made to elucidate various resistance mechanisms. In particular, expression of a drug transport protein, which actively transports an anticancer agent out of cancer cells, thereby reducing the amount of intracellular accumulation of the drug, is considered to play an important role in such a resistance mechanism.
P-glycoprotein, in particular, which is a drug transport protein discovered in the 1970s and is encoded by an MDR1 gene, has been considered a potent target molecule of a multidrug-resistance-overcoming agent, since this protein causes cross-resistance to a plurality of anticancer agents having different chemical structures and action mechanisms. However, it has been gradually elucidated that the anticancer agent resistance mechanism cannot be analyzed on the sole basis of P-glycoprotein, and demand has arisen for development of a resistance-overcoming agent which targets another drug transport protein.
Under such circumstances, there was discovered, in 1998, breast cancer resistance protein (BCRP, also called ABCG2, MXR, or ABCP), which is a drug transport protein belonging to a group which is called “ATP-binding cassette (ABC) transporter superfamily” to which P-glycoprotein also belongs (see Non-Patent Document 1). BCRP has a structure including only one ATP-binding cassette, which differs from that of P-glycoprotein or another ABC transporter, which has two ATP-binding cassettes. BCRP is involved in the mechanism of resistance to a topoisomerase I inhibitor (e.g., irinotecan hydrochloride (CPT-11) or topotecan), to a topoisomerase II inhibitor (e.g., mitoxantrone), or to a molecule-targeting therapeutic drug (e.g., gefitinib and imatinib). Meanwhile, BCRP has been elucidated to exhibit substrate specificity different from that of P-glycoprotein, since BCRP does not act on, for example, paclitaxel or vincristine, which is excreted by P-glycoprotein, and BCRP is involved in excretion of a camptothecin derivative (e.g., CPT-11 or 7-ethyl-10-hydroxycamptothecin (SN-38: active metabolite of CPT-11), which is barely excreted extracellularly by P-glycoprotein (see Non-Patent Document 2). In addition, BCRP has been suggested to be involved in the limitation of the bioavailability of an orally administered anticancer agent (see Non-Patent document 3). In view of the foregoing, demand has arisen for development of a BCRP inhibitor, which is envisaged to exhibit the effect of overcoming anticancer agent resistance that is not overcome by a conventional resistance-overcoming agent, and to improve the bioavailability of an anticancer agent.
Hitherto, in an attempt to overcome resistance to anticancer agents, a variety of P-glycoprotein inhibitors have been developed. In contrast, only a few reports have been given for BCRP inhibitors, and the reported inhibitory action is not satisfactory. Therefore, continuous efforts have been made to develop more effective BCRP inhibitors. Examples of the compounds exhibiting BCRP inhibitory action which have heretofore reported include an FTC (Fumitremorgin C) derivative (see Non-Patent Document 4), estrogen and anti-estorgen (see Non-Patent Document 5), and novobiocin (see Non-Patent Document 6). The present inventors also found that a flavonoid (see Patent Document 1), a diphenylacrylonitrile derivative (see Patent Document 2), and an acrylonitrile derivative having a heterocyclic ring (see Patent Document 3) have BCRP inhibitory action. However, the effects of these compounds are unsatisfactory, and problems in use as pharmaceuticals such as solubility and toxicity have not yet been solved.
Patent Document 1: WO 2004/069233, pamphlet
Patent Document 2: WO 2004/069243, pamphlet
Patent Document 3: WO 2006/106778, pamphlet
Non-Patent Document 1: Proc. Natl. Acad. Sci. USA, 1998, 95: 15665-15670
Non-Patent Document 3: J. Clin. Oncol., 2002, 20: 2943-2950
Non-Patent Document 4: Mol. Cancer. Ther., 2002, 1: 417-425
Non-Patent Document 5: Mol. Cancer. Ther., 2003, 2: 105-112
Non-Patent Document 6: Int. J. Cancer, 2004, 108: 146-151
An object of the present invention is to provide a novel drug which has excellent breast cancer resistance protein (BCRP) inhibitory action and improved solubility.
In an attempt to solve the aforementioned problems, the present inventors have carried out screening of compounds by use of cancer cells which have acquired anticancer drug resistance through BCRP expression, and have found that acrylonitrile derivatives represented by the following formula (1) exhibit potent BCRP inhibitory action.
Accordingly, the present invention provides an acrylonitrile derivative represented by formula (1):
(wherein A represents an optionally substituted 3- to 8-membered heterocyclic ring) or a salt thereof.
The present invention also provides a drug containing, as an active ingredient, the acrylonitrile derivative or a salt thereof as described above.
The present invention also provides a BCRP inhibitor containing, as an active ingredient, the acrylonitrile derivative or a salt thereof as described above.
The present invention also provides an agent for overcoming anticancer agent resistance or an agent for potentiating anticancer agent effect, which agent contains, as an active ingredient, the acrylonitrile derivative or a salt thereof as described above.
The present invention also provides a pharmaceutical composition comprising the acrylonitrile derivative or a salt thereof as described above and a pharmaceutically acceptable carrier.
The present invention also provides an anticancer agent composition comprising the acrylonitrile derivative or a salt thereof as described above and an anticancer agent which serves as a BCRP substrate.
The present invention also provides use of the acrylonitrile derivative or a salt thereof as described above for producing an agent for overcoming anticancer agent resistance or an agent for potentiating anticancer agent effect.
The present invention also provides use of the acrylonitrile derivative or a salt thereof as described above for producing an anticancer agent composition comprising an anticancer agent which serves as a BCRP substrate.
The present invention also provides a method for treatment of cancer which has acquired drug resistance by the mediation of BCRP, the method comprising administering, to a subject in need thereof, the acrylonitrile derivative or a salt thereof as described above.
The present invention also provides a method for inhibiting BCRP, the method comprising administering, to a subject in need thereof, the acrylonitrile derivative or a salt thereof as described above.
The present invention also provides a method for overcoming anticancer agent resistance or for potentiating anticancer agent effect, the method comprising administering, to a subject in need thereof, the acrylonitrile derivative or a salt thereof as described above.
The present invention also provides a method for treatment of cancer, the method comprising administering, to a subject in need thereof, the acrylonitrile derivative or a salt thereof as described above and an anticancer agent which serves as a BCRP substrate.
According to the present invention, the BCRP inhibitory effect of the acrylonitrile derivative or a salt thereof can overcome BCRP-related resistance to an anticancer agent. In addition, the effect of an anticancer agent with respect to cancer cells in which BCRP is intrinsically expressed can be potentiated. Furthermore, according to the present invention, bioavailability of an anticancer agent is envisaged to be enhanced, leading to improvement in the performance of cancer chemotherapy.
The acrylonitrile derivative or a salt thereof of the present invention, which has excellent solubility, is a very useful raw material for producing drugs.
In formula (1), examples of the heterocyclic ring A include 3- to 8-membered heterocyclic rings. Of these, 4- to 8-membered heterocyclic rings are preferred. Specific examples of the heterocyclic ring A include aziridine, azetidine, pyrrolidine, piperidine, piperazine, thiomorpholine, morpholine, azepane, and azocane. Among them, the heterocyclic rings represented by formulas (2) to (9) are preferred, with piperidine represented by formula (5) being particularly preferred.
In formula (1), the heterocyclic ring A may further have one or more substituents. Examples of such substituents include a hydroxyl group, a lower alkyl group, a lower alkoxy group, a lower acyloxy group, a nitro group, an amino group, a halogen atom, a lower hydroxyalkyl group, and a lower alkoxycarbonyl group. When a plurality of substituents are present, those may be identical to or different from one another. Among them, heterocyclic rings having a hydroxyl group are particularly preferred.
Examples of the lower alkyl group include C1 to C6 alkyl groups. Specific examples include methyl, ethyl, n-propyl, and isobutyl.
Examples of the lower alkoxy group include C1 to C6 alkoxy groups. Specific examples include methoxy and ethoxy.
Examples of the lower acyloxy group include C1 to C6 acyloxy groups. Specific examples include formyloxy, acetoxy, and propionyloxy.
Examples of the lower hydroxyalkyl group include C1 to C6 hydroxyalkyl groups. Specific examples include hydroxymethyl and hydroxyethyl.
Examples of the lower alkoxycarbonyl group include C1 to C6 alkoxycarbonyl groups. Specific examples include methoxycarbonyl and ethoxycarbonyl.
Examples of the halogen atom include chlorine, bromine, fluorine, and iodine.
The acrylonitrile derivatives of the present invention may form pharmaceutically acceptable salts thereof, and these salts also fall within the scope of the present invention. Examples of the salts include inorganic salts such as hydrochlorides, sulfates, nitrates, and phosphates; alkali metal salts such as sodium salts and potassium salts; alkaline earth metal salts such as calcium salts and magnesium salts; and organic acid salts such as p-toluenesulfonates, methanesulfonates, fumarates, succinates, and lactates. The compounds of the present invention maybe present in the form of solvate (hydrate). The hydrates also fall within the scope of the present invention. The acrylonitrile derivatives of the present invention may include isomers thereof, and each of these isomers and mixtures of the isomers also fall within the scope of the present invention.
Of these, particularly preferred are the following compounds:
The acrylonitrile derivatives of the present invention and salts thereof may be produced through, for example, the following reaction scheme (A).
Specifically, through condensation between 5-(4-hydroxypiperidin-1-yl)-thiophene-2-carboxaldehyde (X) and 3,4-dimethoxybenzyl cyanide (Y), (Z)-2-(3,4-dimethoxy-phenyl)-3-[5-(4-hydroxy-piperidin-1-yl)-thiophen-2-yl]-acrylonitrile (Z) is produced.
Further reaction of (Z) with bromoacetyl bromide (XX) yields bromo-acetic acid 1-[5-[(Z)-2-cyano-2-(3,4-dimethoxy-phenyl)-vinyl]-thiophen-2-yl]-piperidin-4-yl ester (XY). Reaction of bromo-acetic acid 1-[5-[(Z)-2-cyano-2-(3,4-dimethoxy-phenyl)-vinyl]-thiophen-2-yl]-piperidin-4-yl ester (XY) with a secondary amine of interest (XZ, wherein A has the same meaning as defined above) yields an acrylonitrile derivative of the present invention represented by formula (1) (wherein A has the same meaning as defined above).
No particular limitation is imposed on the secondary amine (XZ, wherein A has the same meaning as defined above). Specific examples thereof include piperidine, 4-hydroxypiperidine, aziridine-2-carboxylic acid methyl ester, azetidine, pyrrolidine, 2-methyl-piperidine, 3-hydroxymethyl-piperidine, azepane (hexamethyleneimine), azocane (heptamethyleneimine), 4-methyl-piperazine, morpholine, and thiomorpholine. Examples of the acrylonitrile derivative produced by use of the secondary amine in the above reaction scheme (A) include:
The condensation reaction is preferably carried out in the presence of a base such as sodium alkoxide, sodium hydroxide, or potassium hydroxide. When sodium alkoxide is employed, the condensation reaction is performed in an alcoholic solvent, such as methanol or ethanol, at between ice cooling temperature and reflux temperature, whereas when sodium hydroxide is employed, the condensation reaction is performed in a solvent mixture of water and an inert solvent, such as methylene chloride or chloroform, with a quaternary ammonium salt or a similar compound being added thereto.
The acrylonitrile derivatives each having a heterocyclic ring of the present invention or salts thereof may be administered as is. Alternatively, the derivatives or salts thereof may be mixed with a pharmaceutically acceptable carrier such as a dispersing aid or an excipient, and may be used in the form of an injection or a peroral preparation such as powder, solution, capsules, suspension, emulsion, syrup, elixir, granules, pills, tablets, troches, or lemonade. These products may be prepared through a conventional method.
Examples of such a carrier include water-soluble monosaccharides, oligosaccharides, and polysaccharides, such as mannitol, lactose, and dextran; gel-forming or water-soluble celluloses, such as hydroxypropyl cellulose, hydroxypropylmethyl cellulose, and methyl cellulose; water-absorbing and poorly water-soluble celluloses, such as crystalline cellulose, α-cellulose, cross-linked carboxymethylcellulose sodium, and derivatives thereof; water-absorbing and poorly water-soluble polysaccharides, such as hydroxypropyl starch, carboxymethyl starch, cross-linked starch, amylose, amylopectin, pectin, and derivatives thereof; water-absorbing and poorly water-soluble gums, such as gum arabic, tragacanth gum, glucomannan, and derivatives thereof; cross-linked vinyl polymers, such as polyvinyl pyrrolidone, cross-linked polyacrylic acid and salts thereof, cross-linked polyvinyl alcohol, polyhydroxyethyl methacrylate, and derivatives thereof; and molecular aggregate (e.g., liposome)-forming lipids, such as phospholipid and cholesterol.
The solubility of the compound of the present invention may further be enhanced through solubilization. Examples of the solubilization technique include techniques which are generally applicable to drugs, such as a technique in which a surfactant (e.g., a polyoxyethylene alcohol ether, a polyoxyethylene acyl ester, a sorbitan acyl ester, or a polyoxyethylene sorbitan acyl ester) is added, and a technique employing a water-soluble polymer (e.g., polyethylene glycol). If desired, there may be employed, for example, a technique for forming a soluble salt of the compound, or a technique for forming a clathrate compound by use of cyclodextrin or a similar material. A solubilization technique may be appropriately selected in accordance with the target acrylonitrile derivative or a salt thereof.
By virtue of a potent BCRP inhibitory effect, the compound of the present invention can be employed as a BCRP inhibitor, an agent for overcoming anticancer agent resistance, and an agent for potentiating anticancer agent effect. The compound of the invention can also be used in an anticancer agent composition in combination with another anticancer agent which serves as a BCRP substrate. The compound may be employed as an agent for overcoming anticancer agent resistance for a cancer which has acquired BCRP-associated resistance through administration of an anticancer drug. Meanwhile, the compound may be employed as an agent for potentiating anticancer agent effect for a cancer which originally expresses BCRP and exhibits low sensitivity to an anticancer drug. No particular limitation is imposed on the target anticancer drug on which the agent for overcoming anticancer agent resistance or agent for potentiating anticancer agent effect containing, as an active ingredient, the BCRP inhibitor of the present invention, acts, so long as the anticancer drug can serve as a substrate for BCRP or an analog thereof. Examples of such an anticancer drug include topoisomerase I inhibitors such as irinotecan hydrochloride/CPT-11 (active form: SN-38) and topotecan; topoisomerase II inhibitors such as mitoxantrone, doxorubicin, daunorubicin, bisanthrene, and etoposide; antifolates such as methotrexate; and molecule-targeting therapeutic drugs such as gefitinib and imatinib. Notably, no particular limitation is imposed on the BCRP analog, so long as it has the same actions on anticancer resistance and sensitivity to anticancer agents as those of BCRP.
The dose of the BCRP inhibitor of the present invention may be appropriately determined in accordance with, for example, the administration method or the symptom of a patient. The daily dose for an adult is preferably 1 mg to 10 g, more preferably 100 mg to 10 g, particularly preferably 500 mg to 10 g. No particular limitation is imposed on the ratio between an anticancer drug and the BCRP inhibitor, and the preferred ratio varies in accordance with, for example, the type of an anticancer drug or inhibitor to be employed. When, for example, irinotecan hydrochloride is employed as an anticancer drug, the ratio by weight of the anticancer drug to the BCRP inhibitor is preferably 1:1 to 1:500, particularly preferably 1:1 to 1:100, more preferably 1:1 to 1:10.
The present invention will next be described in more detail by way of examples, which should not be construed as limiting the invention thereto.
5-Bromothiophene-2-carboxaldehyde was placed in a reactor, and water was added thereto. 4-Hydroxypiperidine (3 eq) was added to the reactor, and the mixture was stirred for tens of minutes or overnight under reflux. Immediately after completion of reaction, the reaction mixture was filtered through filter paper, and the filtrate was cooled for tens of minutes with a flow of water and then for several hours with ice. The precipitated crystals were recovered through filtration under suction and washed with cold water. The crystals were dried and dissolved in chloroform. The chloroform solution was dried over sodium sulfate anhydrate, and the dried solution was filtered through a silica gel pad. The filtrate was washed with chloroform until the color thereof became faint. The filtrate was concentrated under reduced pressure until the start of crystallization. n-Hexane was added to the mixture, and stirring was performed overnight at room temperature. The formed crystals were recovered through filtration, washed with n-hexane, and dried under reduced pressure, to thereby yield 5-(4-hydroxypiperidin-1-yl)-thiophene-2-carboxaldehyde.
5-(4-Hydroxypiperidin-1-yl)-thiophene-2-carboxaldehyde and 3,4-dimethoxybenzyl cyanide were placed in a reactor and dissolved in ethanol. Sodium ethoxide was added thereto, and the mixture was stirred under reflux. After completion of reaction, the reaction mixture was cooled for tens of minutes with a flow of water. Water was added to the cooled mixture, and stirring was performed for tens of minutes. The precipitated crystals were recovered through filtration and washed sequentially with water, ethanol, and hexane, followed by drying under reduced pressure, to thereby yield (Z)-2-(3,4-dimethoxy-phenyl)-3-[5-(4-hydroxy-piperidin-1-yl)-thiophen-2-yl]-acrylonitrile.
Under argon, (Z)-2-(3,4-dimethoxy-phenyl)-3-[5-(4-hydroxy-piperidin-1-yl)-thiophen-2-yl]-acrylonitrile was placed in a reactor and dissolved in chloroform. Pyridine was added to the solution, and the mixture was stirred in an ice bath for tens of minutes. Bromoacetyl bromide was added to the reaction mixture, and stirring was performed in an ice bath for several hours and overnight at room temperature. After completion of reaction, chloroform and water were added to the reaction mixture for phase separation. The recovered organic layer was washed with water and dried over magnesium sulfate anhydrate. The solvent was evaporated under reduced pressure. The residue was purified through silica gel column chromatography, to thereby yield bromo-acetic acid 1-[5-[(Z)-2-cyano-2-(3,4-dimethoxy-phenyl)-vinyl]-thiophen-2-yl]-piperidin-4-yl ester.
(Z)-2-(3,4-dimethoxy-phenyl)-3-[5-(4-hydroxy-piperidin-1-yl)-thiophen-2-yl]-acrylonitrile was placed in a reactor and dissolved in tetrahydrofuran, followed by cooling with ice. Sodium hydride was added to the solution, and the mixture was stirred in an ice bath for tens of minutes. Bromoacetyl chloride was added to the reaction mixture, and stirring was performed for several hours under reflux. After allowing the mixture to stand for cooling, chloroform and water were added thereto for phase separation. The recovered organic layer was washed with water and dried over sodium sulfate anhydrate. The solvent was evaporated under reduced pressure. The residue was purified through silica gel column chromatography, to thereby yield bromo-acetic acid 1-[5-[(Z)-2-cyano-2-(3,4-dimethoxy-phenyl)-vinyl]-thiophen-2-yl]-piperidin-4-yl ester.
Under argon, bromo-acetic acid 1-[5-[(Z)-2-cyano-2-(3,4-dimethoxy-phenyl)-vinyl]-thiophen-2-yl]-piperidin-4-yl ester was placed in a reactor and dissolved in chloroform. The mixture was stirred in an ice bath for tens of minutes. A secondary amine of interest and a tertiary amine of interest (e.g., triethylamine) were added thereto, and stirring was performed in an ice bath for several hours and overnight at room temperature. After completion of reaction, the reaction mixture was washed with water, and the organic layer was dried over sodium sulfate anhydrate. The solvent was evaporated under reduced pressure. The residue was purified through silica gel column chromatography, to thereby yield an acrylonitrile derivative of interest.
Specific examples of production of various derivatives and analytical results of the produced derivatives will next be described.
By use of 5-bromothiophene-2-carboxaldehyde (42.30 g) and 4-hydroxypiperidine (67.30 g), amine incorporation was performed according to Production Step 1, to thereby yield 5-(4-hydroxy-piperidin-1-yl)-thiophene-2-carboxaldehyde (yield: 33.00 g, 71%). The thus-produced 5-(4-hydroxy-piperidin-1-yl)-thiophene-2-carboxaldehyde (10.56 g) and 3,4-dimethoxybenzyl cyanide (8.86 g) were subjected to condensation according to Production Step 2, to thereby yield (Z)-2-(3,4-dimethoxy-phenyl)-3-[5-(4-hydroxy-piperidin-1-yl)-thiophen-2-yl]-acrylonitrile (yield: 13.50 g, 73%). The thus-produced (Z)-2-(3,4-dimethoxy-phenyl)-3-[5-(4-hydroxy-piperidin-1-yl)-thiophen-2-yl]-acrylonitrile (20.00 g) was dissolved in chloroform (650 mL), and the solution was reacted with pyridine (6.41 g) and bromoacetyl bromide (14.13 g) according to Production Step 3 (Method A), to thereby yield bromo-acetic acid 1-[5-[(Z)-2-cyano-2-(3,4-dimethoxy-phenyl)-vinyl]-thiophen-2-yl]-piperidin-4-yl ester (yield: 23.00 g, 87%). The thus-produced bromo-acetic acid 1-[5-[(Z)-2-cyano-2-(3,4-dimethoxy-phenyl)-vinyl]-thiophen-2-yl]-piperidin-4-yl ester (2.30 g) was dissolved in chloroform (100 mL), and the solution was reacted with piperidine (533 mg) and triethylamine (658 mg) according to Production Step 4, to thereby yield the title compound (yield: 1.40 g, 60%).
Bromo-acetic acid 1-[5-[(Z)-2-cyano-2-(3,4-dimethoxy-phenyl)-vinyl]-thiophen-2-yl]-piperidin-4-yl ester (910 mg), produced during the production of Compound 1, was dissolved in chloroform (50 mL), and the solution was reacted with 4-hydroxypiperidine (233 mg) and triethylamine (233 mg) according to Production Step 4, to thereby yield the title compound (yield: 622 mg, 64%).
1H-NMR (CDCl3) δ: 7.37(1H, s), 7.22(1H, d, J=4.4), 7.13(1H, dd, J=8.5, 2.2), 7.04(1H, d, J=2.2), 6.87(1H, d, J=8.5), 6.05(1H, d, J=4.4), 5.08-5.02 (1H, m), 3.94(3H, s), 3.90(3H, s), 3.77-3.70 (1H, m), 3.58-3.52 (2H, m), 3.31-3.27 (2H, m), 3.25(2H, s), 2.87-2.82 (2H, m), 2.42-2.36 (2H, m), 2.43-2.35 (2H, m), 1.95-1.82 (4H, m), 1.70-1.60 (2H, m)
6-mol/L Hydrochloric acid (352 μL), 2-propanol (25 mL), and ethanol (10 mL) were added to Compound 1 (950 mg), and the mixture was heated at about 80° C. for dissolution. After allowing the solution to stand for cooling, the solvent was evaporated under reduced pressure, and the residue was recrystallized from 2-propanol-methanol, to thereby yield the title compound (yield: 869 mg, 85%).
1H-NMR (DMSO-d6) δ: 10.01(1H, br s), 7.94 (1H, s), 7.42(1H, d, J=4.4), 7.17(1H, d, J=2.2), 7.09(1H, dd, J=8.8, 2.2), 7.01(1H, d, J=8.8), 6.32(1H, d, J=4.4), 5.13-5.09 (1H, m), 4.23(2H, br s), 3.83 (3H, s), 3.78(3H, s), 3.56-3.42 (4H, m), 3.40-3.30 (4H, m), 3.05-2.95 (2H, m), 2.08-2.02 (2H, m), 1.85-1.65 (6H, m)
6-mol/L Hydrochloric acid (342 μL), 2-propanol (30 mL), and methanol (30 mL) were added to Compound 2 (956 mg), and the mixture was heated at about 80° C. for dissolution. After allowing the solution to stand for cooling, the solvent was evaporated under reduced pressure, and the residue was recrystallized from 2-propanol, to thereby yield the title compound (yield: 947 mg, 93%).
1H-NMR (DMSO-d6) δ: 10.14(1H, br s), 7.94 (1H, s), 7.42(1H, d, J=4.4), 7.17(1H, d, J=2.0), 7.09(1H, dd, J=8.4, 2.0), 7.01(1H, d, J=8.4), 6.32(1H, d, J=4.4), 5.13-5.07 (1H, m), 4.29-4.23 (2H, m), 3.83(3H, s), 3.78(3H, s), 3.58-3.46 (3H, m), 3.40-3.26 (6H, m), 2.07-1.90 (4H, m), 1.85-1.65 (4H, m)
Methanesulfonic acid (137 μL), 2-propanol (20 mL), and methanol (5 mL) were added to Compound 1 (951 mg), and the mixture was heated at about 80° C. for dissolution. After allowing the solution to stand for cooling, the solvent was evaporated under reduced pressure, and the residue was recrystallized from 2-propanol-methanol, to thereby yield the title compound (yield: 1,078 mg, 95%).
1H-NMR (DMSO-d6) δ: 9.78(1H, br s), 7.93 (1H, s), 7.42(1H, d, J=4.4), 7.17(1H, d, J=2.2), 7.09(1H, dd, J=8.8, 2.2), 7.01(1H, d, J=8.8), 6.32(1H, d, J=4.4), 5.13-5.09(1H, m), 4.24(2H, br s), 3.83 (3H, s), 3.78(3H, s), 3.55-3.40 (4H, m), 3.38-3.30 (4H, m), 3.06-2.94 (2H, m), 2.29(3H, s), 2.08-2.02 (2H, m), 1.85-1.65 (6H, m)
Methanesulfonic acid (144 μL), 2-propanol (20 mL), and methanol (10 mL) were added to Compound 2 (1,029 mg), and the mixture was heated at about 80° C. for dissolution. After allowing the solution to stand for cooling, the solvent was evaporated under reduced pressure, and the residue was recrystallized from 2-propanol-methanol, to thereby yield the title compound (yield: 696 mg, 57%).
1H-NMR (CDCl3) δ: 11.32(1H, br s), 7.37 (1H, s), 7.22(1H, d, J=4.4), 7.13(1H, dd, J=8.5, 2.2), 7.04(1H, d, J=2.2), 6.88(1H, d, J=8.5), 6.06(1H, d, J=4.4), 5.18-5.12 (1H, m), 4.29(1H, br s), 3.95 (3H, s), 3.93(2H, br s), 3.91 (3H, s), 3.65-3.52 (4H, m), 3.50-3.38 (2H, m), 3.33-3.25 (2H, m), 2.78(3H, s), 2.48-2.30 (2H, m), 2.15-2.05 (2H, m), 2.00-1.87 (4H, m), 1.65-1.55 (2H, m)
In a manner similar to that employed in the production of Compound 2, reaction was performed by use of azetidine and triethylamine according to Production Step 4, to thereby yield the title compound (yield: 74%).
1H-NMR (CDCl3) δ: 7.37(1H, s), 7.22(1H, d, J=4.1), 7.13(1H, dd, J=8.3, 2.2), 7.04(1H, d, J=2.2), 6.87(1H, d, J=8.3), 6.05(1H, d, J=4.1), 5.07-4.98 (1H, m), 3.95(3H, s), 3.91(3H, s), 3.59-3.50 (2H, m), 3.35(4H, t-like, J=7.1), 3.31-3.22 (2H, m), 3.26(2H, s), 2.14(2H, quint-like, J=7.1), 2.08-1.97 (2H, m), 1.92-1.79 (2H, m)
Compound 7 (300 mg) was suspended in methanol (2 mL) and dissolved by adding 1-mol/L methanesulfonic acid (in methanol) (630 μL) to the suspension. The solvent was evaporated under reduced pressure, and ether was added the residue to form powder thereof, to thereby yield the title compound (yield: 345 mg, 95%).
MS (FAB) m/z: 468[M+H]+
1H-NMR (CDCl3) δ: 11.80(1H, br s), 7.37 (1H, s), 7.21(1H, d, J=4.1), 7.13(1H, dd, J=8.5, 2.2), 7.03(1H, d, J=2.2), 6.87(1H, d, J=8.5), 6.06(1H, d, J=4.1), 5.17-5.08 (1H, m), 4.63-4.52 (2H, m), 4.15-4.02 (4H, m), 3.94(3H, s), 3.90(3H, s), 3.60-3.50 (2H, m), 3.34-3.24(2H, m), 2.87-2.75 (1H, m), 2.81(3H, s), 2.56-2.41 (1H, m), 2.12-2.02 (2H, m), 1.97-1.85 (2H, m)
In a manner similar to that employed in the production of Compound 2, reaction was performed by use of pyrrolidine and triethylamine according to Production Step 4, to thereby yield the title compound (yield: 74%).
1H-NMR (CDCl3) δ: 7.37(1H, s), 7.22(1H, d, J=4.4), 7.13(1H, dd, J=8.3, 2.2), 7.04(1H, d, J=2.2), 6.87(1H, d, J=8.3), 6.05(1H, d, J=4.4), 5.10-5.02 (1H, m), 3.95(3H, s), 3.91(3H, s), 3.62-3.50 (2H, m), 3.37(2H, s), 3.32-3.22 (2H, m), 2.72-2.58 (4H, m), 2.10-1.99 (2H, m), 1.94-1.78 (6H, m)
Compound 9 (300 mg) was suspended in methanol (3 mL) and dissolved by adding 1-mol/L methanesulfonic acid (in methanol) (620 μl) to the suspension. The solution was added to a mixture of ether (50 mL) and 2-propanol (10 mL) under stirring and cooling with ice, to thereby yield the title compound (yield: 342 mg, 95%).
MS (FAB) m/z: 482[M+H]+
1H-NMR (CDCl3) δ: 11.58(1H, br s), 7.37 (1H, s), 7.21(1H, d, J=4.4), 7.12(1H, dd, J=8.5, 2.2), 7.04(1H, d, J=2.2), 6.87(1H, d, J=8.5), 6.06(1H, d, J=4.4), 5.18-5.08 (1H, m), 4.11(2H, s), 4.05-3.91 (2H, br m), 3.94(3H, s), 3.90(3H, s), 3.61-3.49 (2H, m), 3.35-3.22 (2H, m), 3.18-3.01 (2H, br), 2.77(3H, s), 2.27-1.85 (8H, m)
In a manner similar to that employed in the production of Compound 2, reaction was performed by use of 2-methyl-piperidine and triethylamine according to Production Step 4, to thereby yield the title compound (yield: 73%).
1H-NMR (CDCl3) δ: 7.36(1H, s), 7.22(1H, d, J=4.4), 7.13(1H, dd, J=8.3, 2.2), 7.04(1H, d, J=2.2), 6.87(1H, d, J=8.3), 6.05(1H, d, J=4.4), 5.09-5.00 (1H, m), 3.94(3H, s), 3.90(3H, s), 3.60-3.48 (2H, m), 3.41(2H, d, J=4.1), 3.34-3.22 (2H, m), 2.91-2.82 (1H, m), 2.62-2.47 (2H, m), 2.09-1.98 (2H, m), 1.81-1.80 (2H, m), 1.77-1.50 (4H, m), 1.38-1.21 (2H, m), 1.08(3H, d, J=6.3)
Compound 11 (312 mg) was suspended in methanol (2 mL) and dissolved by adding 1-mol/L methanesulfonic acid (in methanol) (610 μL) to the suspension. The solvent was evaporated under reduced pressure, and ether was added the residue to form powder thereof, to thereby yield the title compound (yield: 320 mg, 86%).
MS (FAB) m/z: 510[M+H]
1H-NMR (CDCl3) δ: 10.84(1H, br s), 7.38 (1H, s), 7.22(1H, d, J=4.1), 7.14(1H, dd, J=8.3, 2.0), 7.05(1H, d, J=2.0), 6.88(1H, d, J=8.3), 6.08(1H, d, J=4.1), 5.15-5.06 (1H, m), 4.24(1H, d, J=18.3), 4.09(1H, d, J=18.3), 3.94(3H, s), 3.90 (3H, s), 3.75-3.64 (1H, m), 3.63-3.43 (4H, m), 3.35-3.24 (2H, m), 2.83(3H, s), 2.15-2.03 (3H, m), 1.98-1.79 (6H, m), 1.62-1.48 (1H, m), 1.43(3H, d, J=6.3)
In a manner similar to that employed in the production of Compound 2, reaction was performed by use of 3-hydroxymethyl-piperidine and triethylamine according to Production Step 4, to thereby yield the title compound (yield: 90%).
1H-NMR (CDCl3) δ: 7.36(1H, d, J=0.5), 7.22(1H, dd, J=4.4, 0.5), 7.13(1H, dd, J=8.5, 2.2), 7.04 (1H, d, J=2.2), 6.87(1H, d, J=8.5), 6.05(1H, d, J=4.4), 5.09-5.00 (1H, m), 3.94(3H, s), 3.90(3H, s), 3.67-3.49 (4H, m), 3.33-3.22 (2H, m), 3.23(2H, d, J=5.1), 2.90-2.80 (1H, m), 2.78-2.67 (1H, m), 2.39-2.17 (2H, m), 2.10-1.97 (2H, m), 1.93-1.55 (7H, m), 1.22-1.08 (1H, m)
Compound 13 (330 mg) was suspended in methanol (3 mL) and dissolved by adding 1-mol/L methanesulfonic acid (in methanol) (630 μl) to the suspension. The solvent was evaporated under reduced pressure, and ether was added the residue to form powder thereof, to thereby yield the title compound (yield: 349 mg, 89%).
MS (FAB) m/z: 526[M+H]
1H-NMR (CDCl3) δ: 10.42(1H, br s), 7.37 (1H, s), 7.21(1H, d, J=4.1), 7.12(1H, dd, J=8.5, 2.0), 7.04(1H, d, J=2.0), 6.87(1H, d, J=8.5), 6.10(1H, d, J=4.1), 5.19-5.10 (1H, m), 4.12(1H, d, J=16.8), 4.01(1H, d, J=16.8), 3.94(3H, s), 3.90 (3H, s), 3.81-3.65 (2H, m), 3.61-3.50 (2H, m), 3.47-3.38 (1H, m), 3.35-3.25 (2H, m), 3.20-3.05 (4H, m), 3.02-2.89 (2H, m), 2.80(3H, s), 2.14-1.78 (7H, m)
In a manner similar to that employed in the production of Compound 2, reaction was performed by use of hexamethyleneimine and triethylamine according to Production Step 4, to thereby yield the title compound (yield: 75%).
1H-NMR (CDCl3) δ: 7.36(1H, s), 7.22(1H, d, J=4.4), 7.13(1H, dd, J=8.5, 2.2), 7.04(1H, d, J=2.2), 6.87(1H, d, J=8.5), 6.05(1H, d, J=4.4), 5.08-5.00 (1H, m), 3.94(3H, s), 3.90(3H, s), 3.60-3.50 (2H, m), 3.41(2H, s), 3.23-3.22 (2H, m), 2.82-2.71 (4H, m), 2.09-1.98 (2H, m), 1.92-1.80 (2H, m), 1.72-1.55 (8H, m)
Compound 15 (300 mg) was suspended in methanol (2 mL) and dissolved by adding 1-mol/L methanesulfonic acid (in methanol) (590 μL) to the suspension. The solvent was evaporated under reduced pressure, and ether was added the residue to form powder thereof, to thereby yield the title compound (yield: 327 mg, 92%).
MS (FAB) m/z: 510[M+H]
1H-NMR (CDCl3) δ: 11.10(1H, br s), 7.37 (1H, s), 7.22(1H, d, J=4.1), 7.13(1H, dd, J=8.5, 2.2), 7.04(1H, d, J=2.2), 6.88(1H, d, J=8.5), 6.07(1H, d, J=4.1), 5.17-5.09 (1H, m), 4.11(2H, d, J=3.2), 3.94(3H, s), 3.90(3H, s), 3.67-3.50 (4H, m), 3.42-3.24 (4H, m), 2.82(3H, s), 2.20-2.03 (4H, m), 1.98-1.82 (6H, m), 1.74-1.61 (2H, m)
In a manner similar to that employed in the production of Compound 2, reaction was performed by use of heptamethyleneimine and triethylamine according to Production Step 4, to thereby yield the title compound (yield: 71%).
1H-NMR (CDCl3) δ: 7.37(1H, s), 7.22(1H, d, J=4.1), 7.13(1H, dd, J=8.5, 2.2), 7.04(1H, d, J=2.2), 6.87(1H, d, J=8.5), 6.05(1H, d, J=4.1), 5.08-5.00 (1H, m), 3.95(3H, s), 3.91(3H, s), 3.60-3.50 (2H, m), 3.39(2H, s), 3.34-3.24 (2H, m), 2.77-2.65 (4H, br), 2.09-1.99 (2H, m), 1.92-1.80 (2H, m), 1.68-1.50 (10H, m)
Compound 17 (300 mg) was suspended in methanol (2 mL) and dissolved by adding 1-mol/L methanesulfonic acid (in methanol) (570 μL) to the suspension. The solvent was evaporated under reduced pressure, and ether was added the residue to form powder thereof, to thereby yield the title compound (yield: 327 mg, 92%).
MS (FAB) m/z: 524 [M+H]+
1H-NMR (CDCl3) δ: 10.60(1H, br s), 7.37 (1H, s), 7.22(1H, d, J=4.1), 7.13(1H, dd, J=8.5, 2.2), 7.04(1H, d, J=2.2), 6.87(1H, d, J=8.5), 6.07(1H, d, J=4.1), 5.18-5.09 (1H, m), 4.12(2H, d, J=2.9), 3.94(3H, s), 3.90(3H, s), 3.69-3.50 (4H, m), 3.44-3.22 (4H, m), 2.80(3H, s), 2.18-1.82 (10H, m), 1.78-1.53 (4H, m)
In a manner similar to that employed in the production of Compound 2, reaction was performed by use of 4-methyl-piperazine and triethylamine according to Production Step 4, to thereby yield the title compound (yield: 69%).
1H-NMR (CDCl3) δ: 7.37(1H, s), 7.22(1H, d, J=4.1), 7.13(1H, dd, J=8.5, 2.2), 7.04(1H, d, J=2.2), 6.88(1H, d, J=8.5), 6.05(1H, d, J=4.1), 5.08-5.00 (1H, m), 3.95(3H, s), 3.91(3H, s), 3.60-3.50 (2H, m), 3.33-3.21 (2H, m), 3.24(2H, s), 2.75-2.36 (8H, br m), 2.30(3H, s), 2.09-1.99 (2H, m), 1.92-1.81 (2H, m)
Compound 19 (270 mg) was suspended in methanol (2 mL) and dissolved by adding 1-mol/L methanesulfonic acid (in methanol) (530 μL) to the suspension. The solvent was evaporated under reduced pressure, and ether was added the residue to form powder thereof, to thereby yield the title compound (yield: 285 mg, 89%).
MS (FAB) m/z: 511[M+H]
1H-NMR (CDCl3) δ: 11.60-10.80 (1H, br), 7.37(1H, s), 7.22(1H, d, J=4.1), 7.13(1H, dd, J=8.5, 2.2), 7.04(1H, d, J=2.2), 6.88(1H, d, J=8.5), 6.06(1H, d, J=4.1), 5.10-5.00 (1H, m), 3.94(3H, s), 3.90(3H, s), 3.62-3.49 (4H, m), 3.41(2H, s), 3.34-3.21 (2H, m), 3.18-3.00 (6H, m), 2.90(3H, s), 2.82(3H, s), 2.10-1.99 (2H, m), 1.93-1.81 (2H, m)
In a manner similar to that employed in the production of Compound 2, reaction was performed by use of morpholine and triethylamine according to Production Step 4, to thereby yield the title compound (yield: 73%).
1H-NMR (CDCl3) δ: 7.37(1H, s), 7.22(1H, d, J=4.1), 7.13(1H, dd, J=8.5, 2.2), 7.04(1H, d, J=2.2), 6.87(1H, d, J=8.5), 6.05(1H, d, J=4.1), 5.10-5.00 (1H, m), 3.94(3H, s), 3.90(3H, s), 3.80-3.70 (4H, m), 3.60-3.50 (2H, m), 3.34-3.20 (2H, m), 3.24(2H, s), 2.65-2.54 (4H, m), 2.10-1.98 (2H, m), 1.93-1.80 (2H, m)
Compound 21 (300 mg) was suspended in methanol (2 mL) and dissolved by adding 1-mol/L methanesulfonic acid (in methanol) (600 μL) to the suspension. The solution was added to ether (50 mL) under stirring and cooling with ice, to thereby yield the title compound (yield: 340 mg, 95%).
MS (FAB) m/z: 498 [M+H]
1H-NMR (CDCl3) δ: 13.00-11.00 (1H, br), 7.37(1H, s), 7.21(1H, d, J=4.1), 7.12(1H, dd, J=8.5, 2.2), 7.04(1H, d, J=2.2), 6.87(1H, d, J=8.5), 6.06(1H, d, J=4.1), 5.19-5.10 (1H, m), 4.19-3.97 (4H, m), 4.04(2H, s), 3.94(3H, s), 3.90(3H, s), 3.80-3.62 (2H, br), 3.58-3.49 (2H, m), 3.38-3.17 (4H, m), 2.78(3H, s), 2.13-2.03 (2H, m), 1.98-1.86 (2H, m)
To an inguinal region of each of 6-week-old BALB/c male nude mice (5 mice/group), BCRP-gene-transfected human colon cancer HCT116 cells (HCT116/BCRP cells) (obtained from Dr. Yoshikazu Sugimoto, The Cancer Chemotherapy Center of Japanese Foundation for Cancer Research) were subcutaneously transplanted (2×106 cells/0.1 mL/mouse). When the tumor volume as estimated from (1/2)ab2 (a: longer tumor diameter, b: shorter tumor diameter) reached about 100 to 150 mm3, each of the compounds of the present invention shown in Tables 1 to 4 and a positive control compound (i.e., Compound 14 ((Z)-2-(3,4-dimethoxy-phenyl)-3-{5-[4-(2-hydroxy-ethyl)-piperazin-1-yl]-thiophen-2-yl}-acrylonitrile hydrochloride)), disclosed in Patent Document 3 (WO 2006/106778)) (hereinafter referred to as Earlier Application Compound 14) was orally administered once a day for nine days (9 times in total). Separately, CPT-11 (45 mg/kg/day) was intravenously administered once a day to the mice on day 1, day 5, and day 9 (3 times in total). The dose of each of the compounds of the present invention was adjusted to 0.20 or 0.40 mmol/kg/day, and that of the positive control compound was adjusted to 0.23 or 0.46 mmol/kg/day. For administration, Compound 3 or 5 of the present invention was dissolved in 5% glucose solution, and Compound 4 or 6 was dissolved in Britton-Robinson buffer. Earlier Application Compound 14 was dissolved in 0.1% Tween 80 solution, and CPT-11 was dissolved in physiological saline. A solvent was solely administered to a control group. On day 21 from start of administration, a tumor was extirpated from each mouse and weighed, and the tumor growth inhibition ratio IR (%) was derived from the following equation:
Tumor growth inhibition ratio IR (%)=(1−average tumor weight of each administration group/average tumor weight of control group)×100.
The results are shown in Tables 1 to 4.
As is clear from Tables 1 to 3, the acrylonitrile derivatives of the present invention potently inhibit BCRP also in vivo, exhibiting a remarkably excellent effect of overcoming resistance to anticancer agent.
As compared with Earlier Application Compound 14 (see; Table 4), which was previously confirmed to exhibit an excellent effect of overcoming resistance to anticancer agent in vivo, the compounds of the present invention were found to exhibit further more excellent effect of overcoming resistance to an anticancer agent.
Each (10 mg) of the compounds of the present invention and Earlier Application Compound 14 shown in Table 5 was weighed and placed in a sealable container, and water or 5% glucose solution was added to the container in such an amount as to adjust the final concentration to 100 mg/mL. The contents of the container were stirred at room temperature for about 30 minutes, followed by centrifugation. The compound concentration of the supernatant recovered through centrifugation was quantified through high-performance liquid chromatography. Table 5 shows the results. Some compounds falling within the scope of the present invention (Compounds 5, 12, 14, and 20), and Earlier Application Compound 14 were analyzed in terms of solubility in physiological saline, in a manner similar to that employed in quantification of the compound concentration of the supernatant, except that physiological saline was used as a solvent instead of water or glucose.
As compared with Earlier Application Compound 14, the compounds of the present invention (Compounds 5, 8, 10, 12, 14, 16, 18, 20, and 22) exhibited remarkably high solubility in water and 5% glucose solution. Earlier Application Compound 14 was found to have poor solubility in physiological saline (<1 mg/mL), whereas the compounds of the present invention (Compounds 5, 12, 14, and 20) were found to have high solubility (100 mg/mL) not depending on the composition of the solvent.
Thus, the compounds falling within the scope of the present invention were found to have advantageous characteristics in that the compounds have considerably high solubility, and that use thereof is not limited by the composition of transfusion, in particular, upon administration to patients.
The following ingredients were mixed and tabletted.
| Number | Date | Country | Kind |
|---|---|---|---|
| 2007-312176 | Dec 2007 | JP | national |
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/JP2008/003557 | 12/2/2008 | WO | 00 | 6/3/2010 |
