Analgesic drugs using Nr4a1 antagonists

Abstract

The present invention pertains to: a compound represented by formula (I) (in the formula, R1 represents a hydroxyl group, a mercapto group, a C1-6-alkoxy group, a C1-6-haloalkoxy group, an amino group, a C1-6-alkyl amino group, or a di-C1-6-alkyl amino group, R2 represents a hydroxyl group, a mercapto group, a C1-6-alkoxy group, a C1-6-haloalkoxy group, an amino group, a C1-6-alkyl amino group, or a di-C1-6-alkyl amino group, R3 represents a halogen atom, a nitro group, a carboxyl group, a C1-6-alkoxy-carbonyl group, or a carboxy-C1-6-alkyl group, and R4 represents a substituted or unsubstituted C3-12-hydrocarbon group) or a salt thereof, or a solvate thereof; and an analgesic drug that contains the compound or a salt thereof, or a solvate thereof.

Description

[Technical field]

[0001] The present invention relates to an analgesic drug using an Nr4a1 antagonist. [Background technology]

[0002] Cancer has been the leading cause of death among Japanese people since 1981. The number of cancer patients increases with age, and it is said that one in three to four people over the age of 70 is a cancer patient.

[0003] Common side effects of anticancer drugs, such as bone marrow suppression, vomiting, and hair loss, cause cancer patients to suffer. Some anticancer drugs, such as taxanes such as paclitaxel and platinum drugs such as oxaliplatin, cause peripheral neuropathic pain, such as pain, numbness, and coldness in the hands, feet, and lips. Furthermore, physical pain, so-called "cancer pain," is a major problem for cancer patients in addition to mental and social pain. Cancer pain here refers to pain from the primary cancer lesion, pain from bone metastasis, and neuropathic pain associated with the infiltration of cancer cells into the nerves.

[0004] Nearly 70% of cancer patients experience pain. Cancer pain is treated according to the three-step pain relief ladder defined by the WHO, mainly by combining non-opioid analgesics such as nonsteroidal anti-inflammatory drugs (NSAIDs) and acetaminophen with opioid analgesics such as codeine, morphine, and fentanyl. While many patients can achieve pain relief for 80% of cancer pain with the use of appropriate analgesics, opioids are not effective against neuropathic pain caused by the infiltration of cancer cells into the nerves, bone metastasis pain, and gastrointestinal obstruction. In addition, side effects such as constipation, vomiting, dependence, and tolerance caused by opioids further plague patients. Therefore, the development of analgesics that are effective against these types of pain is desired.

[0005] Thus, at present, the administration of two types of drugs, anticancer drugs and opioid analgesics, for cancer and cancer pain, and the strong side effects that accompany them, result in suffering for cancer patients. Therefore, the development of an "anticancer analgesic drug" that has both analgesic and anticancer effects in one drug and has fewer side effects than conventional analgesics and anticancer drugs, could be a groundbreaking treatment that greatly improves the quality of life for cancer patients. However, at present, there is no drug that has both analgesic and anticancer effects in one drug.

[0006] On the other hand, for "Nuclear receptor subfamily 4, group A member 1, Nr4a1," which is one of the nuclear receptors, Patent Document 1 discloses a method for identifying an active substance that regulates the activity and expression of Nr4a1. Patent Document 1 describes the inhibition of cancer cell growth, but does not disclose any analgesic effect.

[0007] The present inventors previously aimed to obtain an Nr4a1 antagonist, and performed multi-step in silico screening consisting of "narrowing down based on 3D pharmacophore", "narrowing down by molecular docking simulation", and "molecular similarity analysis based on chemical properties" based on the complex structure (PDB; 3V3Q) of Nr4a1 protein and known antagonist TMPA, and identified 10 compounds from an existing compound database as novel Nr4a1 antagonist candidate compounds, and reported that one of them, NRA-8, suppresses pain caused by PACAP (Non-Patent Document 1). However, Non-Patent Document 1 does not disclose the structure of any of the compounds, and does not mention anything about their anticancer effects. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] Special Publication No. 2018-536384 [Non-patent literature]

[0009] [Non-Patent Document 1] Serina Iso, Motoyuki Takahara, Hiroaki Goda, Atsuro Miyata, Takashi Kurihara, Naoki Toyooka, Ichiro ▲Taka▼▲Saki (Poster presentation); Creation of nuclear receptor Nr4a1 antagonists for the development of novel analgesics; 90th Annual Meeting of the Japanese Pharmacological Society, March 15-17, 2017, Nagasaki (Nagasaki Brick Hall and others) Summary of the Invention [Problem to be solved by the invention]

[0010] An objective of the present invention is to provide a pain treatment drug that has a mechanism of action different from that of NSAIDs and opioids and has both analgesic and anticancer effects in one drug. [Means for solving the problem]

[0011] In order to solve the above problems, the present inventors searched for molecules common to chronic pain and the proliferation and invasion of cancer cells in order to select target molecules, and identified Nr4a1, a nuclear receptor, as a candidate molecule. Specifically, by comprehensively analyzing gene expression in the dorsal horn of the spinal cord of neuropathic pain model mice and gene expression in various cancer cell lines using the GeneChip microarray method, we were able to identify Nr4a1 mRNA as a gene whose expression is increased in both pain model mice and cancer cells.

[0012] Therefore, the present inventors have developed a compound having the following formula: [ka] Since the Nr4a1 inhibitory effect of NRA-8 shown in is equal to or stronger than the previously reported Nr4a1 antagonists TMPA and DIM-C-pPhOH, we carried out structural development based on the structure of NRA-8 in order to obtain compounds that exhibit stronger activity than NRA-8. In screening using the NBRE-Luc reporter vector (NBRE; Nr4a1 binding DNA domain), we succeeded in obtaining many 1,2,3,4-tetrahydroquinazoline derivatives that exhibit efficacy equal to or greater than that of the original compound NRA-8, thus completing the present invention.

[0013] That is, the gist of the present invention is as follows. (1) The following formula (I): [ka] (In the formula, R 1 is a hydroxyl group, a mercapto group, C 1-6 -alkoxy group, C 1-6 -Haloalkoxy group, amino group, C 1-6 -Alkylamino group or diC 1-6 - an alkylamino group; R 2 is a hydroxyl group, a mercapto group, C 1-6 -alkoxy group, C 1-6 -Haloalkoxy group, amino group, C 1-6 -Alkylamino group or diC 1-6 - an alkylamino group; R 3 is a halogen atom, a nitro group, a carboxyl group, C 1-6 -Alkoxy-carbonyl group or carboxy-C 1-6 -alkyl group; R 4 is a substituted or unsubstituted C 3-12 - is a hydrocarbon group. or a salt thereof, or a solvate thereof. (2) In the formula (I), R 1 and R 2 The compound or salt thereof, or a solvate of the compound or salt according to (1) above, wherein (3) In the formula (I), R 4is substituted or unsubstituted C 3-12 -alkyl group, or substituted or unsubstituted C 7-12 -aralkyl group, or a salt thereof, or a solvate thereof according to (1) or (2) above. (4) An analgesic comprising the compound according to any one of (1) to (3) above, or a salt thereof, or a solvate thereof. (5) The analgesic according to (4) above, which is applied to cancer patients. (6) Following formula (II): [ka] (In the formula, R 1 , R 2 and R 3 is as defined in formula (I) of claim 1; R 4’ is a substituted or unsubstituted C 2-12 - is a hydrocarbon group. 2. An analgesic comprising a compound represented by the formula: (7) The analgesic according to (6) above, which is applied to cancer patients. Effect of the Invention

[0014] The compound of the present invention is a compound having a phenyl group substituted with a halogen atom at the 2-position of a 1,2,3,4-tetrahydroquinazolin-4-one skeleton and a substituted or unsubstituted hydrocarbon group at the 3-position, and is useful as an analgesic, which is a new pain treatment drug with a mechanism of action different from that of NSAIDs and opioids.

[0015] In addition, the compound of the present invention has both analgesic and anticancer effects, making it possible to treat cancer without side effects such as bone marrow suppression caused by conventional anticancer drugs. It also makes it possible to treat pain without adverse effects such as peptic ulcers caused by NSAIDs and constipation caused by opioids. Since it is expected to be effective in treating both cancer and pain, it reduces the need for multiple drug combinations, which can avoid the occurrence of unexpected adverse effects, and is expected to improve the quality of life of patients with intractable cancer pain. [Brief description of the drawings]

[0016] [Figure 1] FIG. 1 shows the results of the reporter assay and the structure of NRA-8. [Diagram 2] FIG. 2 shows the effect of compounds on PACAP-induced long-term allodynic response. [Diagram 3] FIG. 3 shows the effect of NRA-8 on allodynia in a neuropathic pain model mouse. [Figure 4] FIG. 4 shows the effect of NRA-8 on allodynia in a bone cancer pain model mouse. [Diagram 5] FIG. 5 shows the effect of NRA-8 on the proliferation, survival and migration of Panc1 cells. [Figure 6] FIG. 6 shows the effect of existing analgesics on the proliferation of Panc1 cells. [Figure 7] FIG. 7 shows the evaluation of the compounds of the present invention by reporter assay. [Figure 8] FIG. 8 shows the analgesic evaluation of the compounds of the present invention. [Figure 9] FIG. 9 shows the anticancer activity evaluation (cell lines) of the compounds of the present invention. [Figure 10] FIG. 10 shows the anticancer activity evaluation (transplant model) of the compound of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0017] The present invention will be described in detail below.

[0018] In the above formulas (I) and (II), R 1 or R 2 Represented by C 1-6 Examples of the alkoxy group include a methoxy group, an ethoxy group, a propoxy group, an isopropoxy group, a butoxy group, an isobutoxy group, a sec-butoxy group, a tert-butoxy group, a pentyloxy group, an isopentyloxy group, a hexyloxy group, a cyclopropyloxy group, a cyclobutyloxy group, a cyclopentyloxy group, and a cyclohexyloxy group.

[0019] In the above formulas (I) and (II), R 1 or R 2 Represented by C 1-6 An example of a -haloalkoxy group is a trifluoromethoxy group.

[0020] In the above formulas (I) and (II), R 1 or R 2 Represented by C 1-6 Examples of the -alkylamino group include a methylamino group, an ethylamino group, a propylamino group, an isopropylamino group, a butylamino group, an isobutylamino group, a sec-butylamino group, a tert-butylamino group, a pentylamino group, an isopentylamino group, a hexylamino group, a cyclopropylamino group, a cyclobutylamino group, a cyclopentylamino group, and a cyclohexylamino group.

[0021] In the above formulas (I) and (II), R 1 or R 2 Represented by diC 1-6 Examples of the alkylamino group include a dimethylamino group, a diethylamino group, a dipropylamino group, a diisopropylamino group, and a methylethylamino group.

[0022] In the above formulas (I) and (II), R 3 Examples of the halogen atom represented by the formula (I) include a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom.

[0023] In the above formulas (I) and (II), R 3 Represented by C 1-6Examples of the -alkoxy-carbonyl group include a methoxycarbonyl group, an ethoxycarbonyl group, a propoxycarbonyl group, an isopropoxycarbonyl group, a butoxycarbonyl group, an isobutoxycarbonyl group, a sec-butoxycarbonyl group, a tert-butoxycarbonyl group, a pentyloxycarbonyl group, an isopentyloxycarbonyl group, a hexyloxycarbonyl group, a cyclopropyloxycarbonyl group, a cyclobutyloxycarbonyl group, a cyclopentyloxycarbonyl group, and a cyclohexyloxycarbonyl group.

[0024] In the above formulas (I) and (II), R 3 Carboxy-C represented by 1-6 Examples of the alkyl group include a carboxymethyl group, a carboxyethyl group, and a carboxypropyl group.

[0025] In the formula (I), R 4 Represented by C 3-12 Examples of the hydrocarbon group include linear or branched C groups such as a propyl group, an isopropyl group, a butyl group, an isobutyl group, a sec-butyl group, a tert-butyl group, a pentyl group, an isopentyl group, a hexyl group, a heptyl group, an octyl group, a nonyl group, a decyl group, an undecyl group, and a dodecyl group. 3-12 -Alkyl group; cyclopropyl group, cyclobutyl group, cyclopentyl group, cyclohexyl group, cycloheptyl group, etc. 3-12 -Cycloalkyl group; 1-propenyl group, allyl group, 1-butenyl group, 2-butenyl group, pentenyl group, hexenyl group, oleyl group, etc. 3-12 -Alkenyl groups; 1-propynyl, 2-propynyl (propargyl), 3-butynyl, pentynyl, hexynyl, and other C 3-12 -Alkynyl group; C6H5(CH2) n - (where n is an integer from 1 to 6), naphthylmethyl group, naphthylethyl group, etc. 7-12 -aralkyl groups; aromatic hydrocarbon groups such as phenyl and naphthyl groups; bridged ring hydrocarbon groups such as adamantyl groups; and spiro hydrocarbon groups.

[0026] In the formula (II), R 4’ Represented by C 2-12 - The hydrocarbon group is the above-mentioned C 3-12 -Examples include those that contain an ethyl group in addition to the hydrocarbon group.

[0027] The above C 3-12 -Hydrocarbon radicals and C 2-12 The hydrocarbon group is, for example, C 1-6 -alkoxy groups (e.g., methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, isopentyloxy, hexyloxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy), methylenedioxy, C 2-6 -alkenyloxy group, aralkyloxy group (e.g., benzyloxy group, 4-methylbenzyloxy group, 3-methylbenzyloxy group, 2-methylbenzyloxy group, 4-fluorobenzyloxy group, 3-fluorobenzyloxy group, 4-chlorobenzyloxy group, 3-chlorobenzyloxy group), halogen atom (fluorine atom, chlorine atom, bromine atom, iodine atom), C 1-6 -Haloalkyl group, C 1-6 -haloalkoxy groups, acyl groups (e.g., formyl groups, acetyl groups, propanoyl groups, butanoyl groups, pentanoyl groups, hexanoyl groups, etc.) 1-6 -Aliphatic acyl groups; aroyl groups such as benzoyl and toluoyl groups), acyloxy groups (e.g., formyloxy, acetoxy, propanoyloxy, butanoyloxy, pentanoyloxy, and hexanoyloxy groups, etc. 1-6 -Aliphatic acyloxy groups; aroyloxy groups such as benzoyloxy and toluoyloxy groups), which may be substituted with one or more substituents selected from a hydroxyl group, a carboxyl group, an acetamide group, a carbamoyl group, a cyano group, a nitro group, and the like.

[0028] In the above formula (I) or (II), R 4 or R 4’Examples of the substituted hydrocarbon group represented by the formula (I) include a 2-hydroxyethyl group, a 2-methoxyethyl group, a 3-hydroxypropyl group, a 3-methoxypropyl group, a 4-hydroxybutyl group, a 4-methoxybutyl group, a 3-(p-methoxyphenyl)propyl group, a 3-(p-fluorophenyl)propyl group, a 3-(p-chlorophenyl)propyl group, and the like, but are not limited to these.

[0029] The compound represented by the formula (I) or (II) includes R 1 and R 2 is a hydroxyl group, R 3 is a halogen atom, e.g., a chlorine atom, R 4 is substituted or unsubstituted C 3-8 -alkyl group, or substituted or unsubstituted C 7-11 -aralkyl groups are preferred.

[0030] The compound represented by the formula (I) is a novel compound, and the compound represented by the formula (II) includes known compounds in addition to the novel compound represented by the formula (I).

[0031] The salt of the compound represented by formula (I) or (II) is preferably a pharma- ceutically acceptable salt, and examples thereof include salts with inorganic acids such as hydrochloric acid, sulfuric acid, phosphoric acid, hydrobromic acid, hydroiodic acid, nitric acid, pyrosulfuric acid, and metaphosphoric acid, and salts with organic acids such as citric acid, benzoic acid, acetic acid, propionic acid, fumaric acid, maleic acid, and sulfonic acids (e.g., methanesulfonic acid, p-toluenesulfonic acid, and naphthalenesulfonic acid).

[0032] The solvates of the compound represented by the above formula (I) or (II) or a salt thereof include, for example, hydrates.

[0033] In the formula (I), R 1 and R 2 is a hydroxyl group, R 3 A compound in which is a halogen atom, for example a chlorine atom, can be prepared, for example, as shown below. [ka] (In the formula, R 4 has the same meaning as in formula (I).

[0034] That is, the amine compound (R 4 -NH2) and o-nitrobenzoic acid to convert it to o-nitrobenzoic acid amide, which is then reduced and reacted with 3,4-di(benzyloxy)-5-chlorobenzaldehyde to obtain the benzyl form, which can then be debenzylated to obtain the desired product.

[0035] R 1 and R 2 C 1-6 -alkoxy group or C 1-6 When protection with a benzyl group is not required, such as with a -haloalkoxy group, 3,4-di(C 1-6 -Alkoxy or C 1-6 The target compound can be obtained directly by using (-haloalkoxy)-5-chlorobenzaldehyde.

[0036] The compound represented by the above formula (II) can also be obtained in the same manner as above.

[0037] The product obtained as described above can be purified by a commonly used method, such as column chromatography using silica gel or the like as a carrier, or a recrystallization method using methanol, ethanol, chloroform, dimethyl sulfoxide, n-hexane-ethyl acetate, water, etc. Examples of elution solvents for column chromatography include methanol, ethanol, chloroform, acetone, hexane, dichloromethane, ethyl acetate, and mixed solvents thereof.

[0038] The compound can be formulated as an analgesic and / or anticancer drug by combining with a conventional pharmaceutical carrier. The dosage form is not particularly limited and can be appropriately selected and used as needed, and includes oral preparations such as tablets, capsules, granules, fine granules, powders, sustained release preparations, liquids, suspensions, emulsions, syrups, and elixirs, and parenteral preparations such as injections and suppositories.

[0039] Oral preparations are produced by conventional methods using, for example, starch, lactose, sucrose, mannitol, carboxymethylcellulose, inorganic salts, etc. In addition to these, binders, disintegrants, surfactants, lubricants, flow enhancers, flavorings, colorants, fragrances, etc. may be added as appropriate.

[0040] Examples of binders include starch, dextrin, gum arabic, gelatin, hydroxypropyl starch, methyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl cellulose, crystalline cellulose, ethyl cellulose, polyvinylpyrrolidone, and macrogol.

[0041] Examples of disintegrants include starch, hydroxypropyl starch, sodium carboxymethylcellulose, calcium carboxymethylcellulose, carboxymethylcellulose, low-substituted hydroxypropylcellulose, and the like.

[0042] Examples of the surfactant include sodium lauryl sulfate, soybean lecithin, sucrose fatty acid ester, polysorbate 80, and the like.

[0043] Examples of the lubricant include talc, waxes, hydrogenated vegetable oil, sucrose fatty acid ester, magnesium stearate, calcium stearate, aluminum stearate, polyethylene glycol, and the like.

[0044] Examples of the flow enhancer include light anhydrous silicic acid, dried aluminum hydroxide gel, synthetic aluminum silicate, magnesium silicate, and the like.

[0045] Injections are produced according to conventional methods, and diluents that can be used are generally distilled water for injection, physiological saline, glucose aqueous solution, olive oil, sesame oil, peanut oil, soybean oil, corn oil, propylene glycol, polyethylene glycol, etc. Furthermore, bactericides, preservatives, stabilizers, isotonicity agents, soothing agents, etc. may be added as necessary. From the viewpoint of stability, the injections can be frozen after being filled into vials, etc., and water removed by a conventional freeze-drying technique, and a liquid can be reconstituted from the freeze-dried product just before use. The ratio of the compound of formula (I) or (II) in the injection can vary between 5 and 50% by weight, but is not limited thereto.

[0046] Other parenteral preparations include suppositories for rectal administration, which are prepared according to conventional methods.

[0047] The formulated analgesic and / or anticancer drug may be administered, for example, 1 to 4 times a day for a period of 1 week to 3 months, although this varies depending on the dosage form, route of administration, etc.

[0048] In order for an oral preparation to exert the desired effect, it is appropriate for an adult to take, for example, 0.1 to 1000 mg, preferably 1 to 500 mg, of the compound of formula (I) or (II) in divided doses several times a day, although this varies depending on the age, weight, and severity of the disease of the patient.

[0049] In order for a parenteral preparation to exert the desired effect, it is appropriate to administer, for example, 0.1 to 1000 mg, preferably 1 to 500 mg, of the compound of formula (I) or (II) to an adult patient by intravenous injection, intravenous drip infusion, subcutaneous injection, or intramuscular injection, although this varies depending on the age, body weight, and severity of the disease of the patient.

[0050] The compound of the present invention has both analgesic and anticancer effects, and the difference between the doses required for the analgesic and anticancer effects is small. Therefore, when applied to cancer patients, the need for multiple drug combinations is reduced, and the occurrence of unexpected adverse effects can be avoided, and further, the quality of life of patients with intractable cancer pain can be expected to improve. The compound of the present invention exhibited analgesic effects in neuropathic pain models and bone cancer pain models, and also inhibited the proliferation, survival, and migration of various cancer cells, so that it is expected that it will have therapeutic effects as an analgesic drug for intractable chronic pain and as an anticancer drug for various cancers, even when used alone. In particular, it showed a good analgesic effect on the pain response of bone cancer pain model mice (Figure 8). It has already been reported that opioids are not very effective against the pain response of this model mouse (Minami et al., J Pharmacol Sci., 111(1), 60-72, 2009), and opioids are also said to be ineffective against bone metastasis pain of cancer cells in clinical practice. Therefore, the compound of the present invention is expected to have a therapeutic effect against pain associated with metastatic bone tumors, for which opioids are resistant.

[0051] The 5-year survival rate after cancer diagnosis is 8.9% for pancreatic cancer, 29.5% for lung cancer, and 34.1% for brain and central nervous system cancer, which are significantly lower than other tumors (according to statistics from the National Cancer Research Center). This is due to "recurrence" in which tumors that were not completely removed during surgery grow again, or "metastasis" in which cancer is found in another site, and the low 5-year survival rate indicates that drug treatment for the recurrence and metastasis of these tumors is currently incomplete. The compound of the present invention was able to strongly suppress the proliferation, survival, and migration of various cancer cells, including pancreatic cancer cell Panc1, lung cancer cell line A549, and brain tumor (glioma) cells U87 and U251 (Figures 5 and 9). Furthermore, the compound of the present invention showed a tumor proliferation inhibitory effect when administered orally in a xenograft model (Figure 10). From the above, it is expected that the compound can be used to prevent cancer recurrence and metastasis after tumor removal during surgery, which is expected to lead to an improvement in the 5-year survival rate.

[0052] In recent years, it has been reported that the nuclear receptor Nr4a1, which is the target molecule of the present invention, is involved in "PD-L1 expression" (Karki et al., Cancer Res 80(5), 1011-1023, 2020) and "T cell exhaustion" (Liu et al., Nature 567, 525-529, 2019; Chen et al., Nature 567, 530-534, 2019). It is expected that inhibiting Nr4a1 with the compound of the present invention may enhance cancer immunotherapy through "reduced expression of PD-L1 in cancer cells" and "activation of T cells."

[0053] This specification includes the contents disclosed in the specification and drawings of Japanese Patent Application No. 2020-120069, which is the priority basis of this application. EXAMPLES

[0054] The present invention will be described in more detail below with reference to examples, but the scope of the present invention is not limited to these examples.

[0055] [Example 1] Synthesis of 3-substituted-2-phenyl-1,2,3,4-tetrahydroquinazolin-4-one derivatives [ka]

[0056] Under Ar atmosphere, commercially available amine 2a-i (0.60mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (149mg, 0.78mmol), 1-hydroxybenzotriazole (92mg, 0.60mmol), and triethylamine (0.08mL, 0.60mmol) were added to a dichloromethane solution (3mL) of commercially available o-nitrobenzoic acid (100mg, 0.60mmol) in ice-cooled water, and the mixture was stirred at room temperature for 24 hours. Then, 10% HCl aqueous solution (3mL) was added, and the mixture was extracted with dichloromethane (1mL x 3 times). The organic layers were combined and dried over anhydrous sodium sulfate, and the solvent was removed using a rotary evaporator. The residue was purified by silica gel column chromatography (n-hexane:acetone = 3:1 to 1:1) to obtain amide 3a-i.

[0057] 10% palladium activated carbon (5 mg) was added to an ethyl acetate solution (3 mL) of amide 3a-i (0.48 mmol) at room temperature, and the mixture was then substituted with hydrogen and stirred at room temperature for 2 hours. The catalyst was then removed by celite filtration, and the solvent was removed using a rotary evaporator to obtain aniline. The obtained aniline was used in the next reaction without purification. Under an Ar atmosphere, 5-chloro-3,4-dibenzyloxybenzaldehyde (168 mg, 0.48 mmol) and cyanuric acid chloride (18 mg, 0.10 mmol) were added in sequence at room temperature to an acetonitrile solution (3 mL) of the obtained aniline, and the mixture was stirred at room temperature for 24 hours. The solvent was then removed using a rotary evaporator, and the obtained residue was purified by silica gel column chromatography (n-hexane:acetone = 2:1 to 1:1) to obtain 1,2,3,4-tetrahydroquinazolin-4-one derivative 4a-i.

[0058] 20% palladium hydroxide on activated carbon (3 mg) was added to a methanol solution (3 mL) of 1,2,3,4-tetrahydroquinazolin-4-one derivative 4a-i at room temperature, followed by hydrogen substitution and stirring at room temperature for 4 hours. The catalyst was then removed by celite filtration, and the solvent was distilled off using a rotary evaporator. The resulting residue was purified by silica gel column chromatography (dichloromethane:methanol = 50:1 to 10:1) to obtain the desired NRA-807 to NRA-815.

[0059] 3-Butyl-2-(5-chloro-3,4-dihydroxyphenyl)-1,2,3,4-tetrahydroquinazolin-4-one (NRA-807) 1 H-NMR (400 MHz, DMSO-d6) δ: 0.87 (3H, t, J = 7.4 Hz), 1.32 (2H, sext, J = 7.4 Hz), 1.46-1.64 (2H, m), 2.75-2.82 (1H, m), 3.94-4.04 (1H, m), 5.78 (1H, d, J = 2.4 Hz), 6.41 (1H, s), 6.65 (1H, d, J = 7.6 Hz), 6.70 (1H, td, J = 7.6, 1.3 Hz), 6.82 (1H, d, J = 2.4 Hz), 6.90 (1H, d, J = 2.4 Hz), 7.19 (1H, td, J = 7.6, 1.3 Hz), 7.75 (1H, dd, J = 7.6, 1.3 Hz), 8.49 (1H, s)

[0060] 2-(5-chloro-3,4-dihydroxyphenyl)-3-isopropyl-1,2,3,4-tetrahydroquinazolin-4-one (NRA-808) 1H-NMR (400 MHz, DMSO-d6) δ: 1.00 (3H, d, J = 6.9 Hz), 1.26 (3H, d, J = 6.9 Hz), 4.73 (1H, sept, J = 6.9 Hz), 5.82 (1H, d, J = 2.0 Hz), 6.40 (1H, s), 6.58 (1H, d, J = 7.6 Hz), 6.73 (1H, t, J = 7.6 Hz), 6.81 (1H, d, J = 2.0 Hz), 6.89 (1H, d, J = 2.0 Hz), 7.14 (1H, td, J = 7.6, 2.0 Hz), 7.43 (1H, dd, J = 7.6, 2.0 Hz), 8.43 (1H, br) 3-Benzyl-2-(5-chloro-3,4-dihydroxyphenyl)-1,2,3,4-tetrahydroquinazolin-4-one (NRA-809) 1 H-NMR (400 MHz, DMSO-d6) δ: 3.78 (1H, d, J = 15.6 Hz), 5.48 (1H, d, J = 15.6 Hz), 5.63 (1H, d, J = 1.8 Hz), 6.37 (1H, s), 6.67 (1H, d, J = 7.2 Hz), 6.75 (1H, t, J = 7.2 Hz), 6.79 (1H, d, J = 1.8 Hz), 6.83 (1H, d, J = 1.8 Hz), 7.20-7.32 (6H, m), 7.84 (1H, dd, J = 7.2, 1.8 Hz), 8.48 (1H, br)

[0061] 2-(5-chloro-3,4-dihydroxyphenyl)-3-phenethyl-1,2,3,4-tetrahydroquinazolin-4-one (NRA-810) 1H-NMR (400 MHz, DMSO-d6) δ: 2.76-2.79 (1H, m), 2.92-3.07 (2H, m), 4.05-4.11 (1H, m), 5.71 (1H, d, J = 2.0 Hz), 6.34 (1H, s), 6.66 (1H, d, J = 7.5 Hz), 6.72 (1H, t, J = 7.5 Hz), 6.82 (1H, d, J = 2.0 Hz), 6.89 (1H, d, J = 2.0 Hz), 7.15-7.28 (6H, m), 7.79 (1H, dd, J = 7.5, 2.0 Hz), 8.54 (1H, br)

[0062] 2-(5-chloro-3,4-dihydroxyphenyl)-3-(3-phenylpropyl)-1,2,3,4-tetrahydroquinazolin-4-one (NRA-811) 1 H-NMR (400 MHz, DMSO-d6) δ: 1.81-2.00 (2H, m), 2.62 (2H, t, J = 7.6 Hz), 2.84-2.91 (2H, m), 3.92-3.99 (1H, m), 5.78 (1H, d, J = 1.8 Hz), 6.66 (1H, d, J = 7.7 Hz), 6.71 (1H, t, J = 7.7 Hz), 7.13 (1H, t, J = 7.7 Hz), 7.17-7.26 (5H, m), 7.78 (1H, d, J = 7.7 Hz), 8.56 (1H, br)

[0063] 2-(5-chloro-3,4-dihydroxyphenyl)-3-(4-phenylbutyl)-1,2,3,4-tetrahydroquinazolin-4-one (NRA-812) 1H-NMR (400 MHz, DMSO-d6) δ: 1.62-1.65 (5H, m), 2.03-2.08 (1H, m), 2.60-2.61 (1H, m), 3.98-4.05 (1H, m), 5.75 (1H, J = 2.7 Hz), 6.64 (1H, J = 7.7 Hz), 6.79 (1H, d, J = 1.8 Hz), 6.87 (1H, d, J = 1.8 Hz), 7.10-7.25 (6H, m), 7.77 (1H, dd, J = 7.7, 1.8 Hz), 8.52 (2H, br)

[0064] 2-(5-chloro-3,4-dihydroxyphenyl)-3-(3-(p-methoxyphenyl)propyl)-1,2,3,4-tetrahydroquinazolin-4-one (NRA-813) 1 H-NMR (400 MHz, Acetone-d6) δ: 1.79-1.98 (2H, m), 2.55 (2H, t, J = 8.0 Hz), 2.82-2.90 (1H, m), 3.73 (3H, s), 3.88-3.96 (1H, m), 5.77 (1H, d, J = 1.6 Hz), 6.66 (1H, d, J = 8.0 Hz), 6.70 (1H, t, J = 8.0 Hz), 6.78-6.82 (3H, m), 6.87 (1H, d, J = 2.4 Hz), 7.08 (1H, d, J = 8.0 Hz), 7.20 (1H, td, J = 1.6 Hz), 7.77 (1H, dd, J = 8.0, 1.6 Hz)

[0065] 2-(5-chloro-3,4-dihydroxyphenyl)-3-(3-(p-fluorophenyl)propyl)-1,2,3,4-tetrahydroquinazolin-4-one (NRA-814) 1H-NMR (400 MHz, Acetone-d6) δ: 1.80-2.00 (2H, m), 2.63 (2H, t, J = 8.0 Hz), 2.82-2.92 (1H, m), 3.91-3.98 (1H, m), 5.78 (1H, d, J = 2.4 Hz), 6.66 (1H, d, J = 8.0 Hz), 6.72 (1H, t, J = 8.0 Hz), 6.81 (1H, d, J = 2.4 Hz), 6.88 (1H, d, J = 2.4 Hz), 7.00 (2H, t, J = 8.0 Hz), 7.19-7.24 (3H, m), 7.79 (1H, dd, J = 8.0, 1.2 Hz), 8.12 (1H, s), 8.76 (1H, s)

[0066] 2-(5-chloro-3,4-dihydroxyphenyl)-3-(3-(p-chlorophenyl)propyl)-1,2,3,4-tetrahydroquinazolin-4-one (NRA-815) 1 H-NMR (400 MHz, Acetone-d6) δ: 1.80-2.00 (2H, m), 2.63 (2H, t, J = 8.0 Hz), 2.83-2.94 (1H, m), 3.88-3.96 (1H, m), 5.79 (1H, d, J = 1.6 Hz), 6.40 (1H, s), 6.67 (1H, t, J = 8.0 Hz), 6.72 (1H, t, J = 8.0 Hz), 6.82 (1H, d, J = 2.4 Hz), 6.88 (1H, d, J = 2.4 Hz), 7.18-7.28 (5H, m), 7.78 (1H, dd, J = 8.0, 1.2 Hz), 8.56 (1H, br)

[0067] [Example 2] Identification of novel Nr4a1 antagonist candidate compounds Aiming to obtain a novel Nr4a1 antagonist, the inventors performed a multi-step in silico screening consisting of "narrowing down based on 3D pharmacophore," "narrowing down by molecular docking simulation," and "molecular similarity analysis based on chemical properties" based on the complex structure (PDB; 3V3Q) of Nr4a1 protein and known antagonist TMPA, and identified 10 compounds (purchased from Namiki Shoji Co., Ltd.) as novel Nr4a1 antagonist candidate compounds from an existing compound database. Then, a primary screening was performed using the NBRE-Luc reporter vector (NBRE; Nr4a1 binding DNA domain). Specifically, 1 × 10 CHO cells were plated in a 96-well plate. 5 The cells were seeded with 10 compounds and incubated for 24 hours. The NBRE-Luc reporter vector and Nr4a1 expression vector were transfected and incubated for 6 hours. After changing the medium, candidate compounds (10 compounds) and previously reported Nr4a1 antagonists TMPA and DIM-C-pPhOH were added at 20 μM and incubated for 18 hours. Luciferin was then added and luminescence was measured using a microplate reader. In this reporter assay, one compound was identified as having antagonistic properties (named NRA-8). The Nr4a1 inhibitory effect of NRA-8 was equal to or stronger than that of previously reported Nr4a1 antagonists TMPA and DIM-C-pPhOH. The results of the reporter assay and the structure of NRA-8 are shown in Figure 1.

[0068] [Example 3] Drug efficacy evaluation using a mouse pain model 1 (mechanical allodynia induced by intrathecal administration of PACAP) A single intrathecal (it) administration of PACAP (100 pmol / 5 μL) to mice caused a long-term (at least 3 months after administration) hypersensitivity to mechanical stimuli (mechanical allodynia) (Yokai et al. Mol. Pain 2016. 12, 1-13.). The onset of this PACAP-induced long-term mechanical hypersensitivity was almost completely suppressed by simultaneous administration of NRA-8 (1 nmol) (Figure 2). The known Nr4a1 antagonists TMPA and DIM-C-pPhOH used as controls also suppressed the onset, but NRA-8 was the most potent (Figure 2). When the solvent (VEH: artificial cerebrospinal fluid containing 0.2% DMSO) was used instead of the above compound, the PACAP-induced mechanical hypersensitivity was not suppressed.

[0069] Male ddY mice (6-12 weeks old) were used, and the threshold for mechanical stimulation was evaluated by the von Frey test. That is, the 50% withdrawal reflex threshold was calculated according to the method of Chaplan et al. (Chaplan et al., J. Neurosci. Meth. 53, 55-63, 1994). Compounds were dissolved in 99.7% DMSO and then diluted with artificial cerebrospinal fluid (final concentration of DMSO was 0.2%).

[0070] [Example 4] Drug efficacy evaluation using mouse pain model 2 (peripheral neuropathic pain model: mechanical allodynia induced by spinal nerve injury) A spinal nerve ligation (SNL) model was used in which the fourth lumbar spinal nerve of male ddY mice (6-12 weeks old) was ligated with silk thread. Drug efficacy was evaluated 14 days after nerve ligation. It administration of NRA-8 (100 pmol and 1 nmol) dose-dependently suppressed mechanical allodynia induced by nerve ligation (Figure 3). On the other hand, when the vehicle (VEH: artificial cerebrospinal fluid containing 0.2% DMSO) was used, mechanical allodynia was not suppressed.

[0071] [Example 5] Drug efficacy evaluation using mouse pain model 3 (bone cancer pain model: mechanical allodynia induced by transplantation of cancer cells into the femur) We investigated the effect of drugs on mechanical allodynia induced by transplanting NCTC2472 cells into the femoral bone marrow (at the knee joint site) of male C3H / HeN mice (6-12 weeks old). NRA-8 was orally administered on the 10th day after cancer cell transplantation. Oral administration of NRA-8 (10 and 30 mg / kg) dose-dependently suppressed mechanical allodynia induced by cancer cell transplantation (Figure 4). On the other hand, when the vehicle (VEH: distilled water containing 0.1% DMSO) was used, mechanical allodynia was not suppressed.

[0072] [Example 6] Anticancer activity against human pancreatic cancer cell line Panc1 When the anticancer activity against the human pancreatic cancer cell line Panc1 was examined, the novel Nr4a1 antagonist NRA-8 inhibited the proliferation, survival, and migration of Panc1 cells (Figure 5). On the other hand, no anticancer activity (inhibitory effect on cell proliferation) was observed in existing analgesics (aspirin (acetylsalicylic acid), diclofenac, and pregabalin) (Figure 6). NRA-8 also showed similar anticancer activity in various cancer cell lines, including A549 cells (human lung cancer), U87 and U251 (human brain tumors), LNCaP (human prostate cancer), and MCF7 (human breast cancer).

[0073] Furthermore, the present inventors carried out structure expansion based on the structure of NRA-8, aiming to obtain a compound showing stronger activity than NRA-8. In screening using the NBRE-Luc reporter vector (NBRE; Nr4a1 binding DNA domain), they succeeded in obtaining NRA-811 as a compound showing efficacy equal to or higher than that of the original compound NRA-8. In addition, they carried out structure expansion of compounds with stronger activity based on the structure of NRA-811, and performed a reporter assay, and succeeded in obtaining compounds NRA-813, 814 and 815 with stronger activity than NRA-811.

[0074] The evaluation of the compounds of the present invention by reporter assay is shown in FIG.

[0075] It showed stronger analgesic effects than NRA-8 in the PACAP-induced long-term pain model, the sciatic nerve ligation neuropathic pain model, and the bone cancer pain model (Figure 8).

[0076] In addition, NRA-811 and 815 suppressed cell proliferation, survival, and migration in the human pancreatic cancer cell line Panc1, and the effects were stronger than those of the parent compound (NRA-8) (Figure 9). NRA-811 and 815 also showed similar anticancer activity in various cancer cell lines, including A549 cells (human lung cancer), U87 and U251 (human brain tumor), LNCaP (human prostate cancer), and MCF7 (human breast cancer).

[0077] [Example 7] Anticancer activity in xenograft model (cancer cell transplant model) Male nude mice (Balb / c nu / nu, 6 weeks old) were used, and the human pancreatic cancer cell line Panc1 (1 × 10 6 The NRA compound was suspended in water containing 0.3% methylcellulose and orally administered once a day for 7 days starting from the 7th day after cancer cell transplantation. The tumor size was measured using a vernier caliper to measure the long and short diameters (mm) of the tumor, and the volume was calculated as long diameter (mm) x short diameter (mm) x short diameter (mm).

[0078] The tumor growth inhibitory effect of NRA-8 was not strong in the xenograft model, but the compounds of the present invention, NRA-811 and 815, exhibited a stronger tumor growth inhibitory effect than NRA-8 (FIG. 10).

[0079] All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety.

Claims

1. The following formula (I): 【Chemistry 1】 (In the formula, R １ is a hydroxyl group, a mercapto group, C １－６ -alkoxy group, C １－６ -Haloalkoxy group, amino group, C １－６ -Alkylamino group or diC １－６ - an alkylamino group; R ２ is a hydroxyl group, a mercapto group, C １－６ -alkoxy group, C １－６ -Haloalkoxy group, amino group, C １－６ -Alkylamino group or diC １－６ - an alkylamino group; R ３ is a halogen atom, a nitro group, a carboxyl group, C １－６ -alkoxy-carbonyl group or carboxy-C １－６ - is an alkyl group; R ４ is a substituted or unsubstituted C ３－１２ - a hydrocarbon group (excluding propyl, isobutyl and allyl groups). or a salt thereof, or a solvate thereof.

2. In the formula (I), R １ and R ２ The compound according to claim 1, or a salt thereof, or a solvate thereof, wherein

3. In the formula (I), R ４ is substituted or unsubstituted C ３－１２ -Alkyl group, or substituted or unsubstituted C ７－１２ 3. The compound according to claim 1 or 2, wherein -aralkyl group, or a salt thereof, or a solvate of the compound or the salt.

4. 4. An analgesic comprising the compound according to any one of claims 1 to 3, or a salt thereof, or a solvate thereof.

5. The analgesic according to claim 4, which is used for cancer patients.

6. The following formula (II): 【Chemistry 2】 (In the formula, R １ , R ２ and R ３ is as defined in formula (I) of claim 1; R ４’ is a substituted or unsubstituted C ２－１２ - is a hydrocarbon group. An analgesic comprising a compound represented by the following formula (I):

7. The analgesic according to claim 6, which is applied to cancer patients.

8. The following formula (I): 【Chemistry 3】 (In the formula, R 1 is a hydroxyl group, a mercapto group, a C 1-6 -alkoxy group, a C 1-6 -haloalkoxy group, an amino group, a C 1-6 -alkylamino group or a diC 1-6 -alkylamino group; R 2 is a hydroxyl group, a mercapto group, a C 1-6 -alkoxy group, a C 1-6 -haloalkoxy group, an amino group, a C 1-6 -alkylamino group or a diC 1-6 -alkylamino group; R 3 is a halogen atom, a nitro group, a carboxyl group, a C 1-6 -alkoxy-carbonyl group or a carboxy-C 1-6 -alkyl group; and R 4 is a substituted or unsubstituted C 3-12 -hydrocarbon group.) An analgesic comprising a compound represented by the following formula (I):

9. The following formula (I): 【Chemistry 4】 (In the formula, R 1 is a hydroxyl group, a mercapto group, a C 1-6 -alkoxy group, a C 1-6 -haloalkoxy group, an amino group, a C 1-6 -alkylamino group or a diC 1-6 -alkylamino group; R 2 is a hydroxyl group, a mercapto group, a C 1-6 -alkoxy group, a C 1-6 -haloalkoxy group, an amino group, a C 1-6 -alkylamino group or a diC 1-6 -alkylamino group; R 3 is a halogen atom, a nitro group, a carboxyl group, a C 1-6 -alkoxy-carbonyl group or a carboxy-C 1-6 -alkyl group; and R 4 is a substituted or unsubstituted C 3-12 -hydrocarbon group.) An anticancer drug comprising a compound represented by the formula:

Images