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WO2025121320A1 - Agent prophylactique ou thérapeutique contre la polykystose rénale - Google Patents

Agent prophylactique ou thérapeutique contre la polykystose rénale Download PDF

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WO2025121320A1
WO2025121320A1 PCT/JP2024/042728 JP2024042728W WO2025121320A1 WO 2025121320 A1 WO2025121320 A1 WO 2025121320A1 JP 2024042728 W JP2024042728 W JP 2024042728W WO 2025121320 A1 WO2025121320 A1 WO 2025121320A1
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Japanese (ja)
Inventor
達雄 宮本
岳志 板橋
知佳 森田
啓司 鈴木
卓 山本
康介 細羽
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Hiroshima University NUC
Nagasaki University NUC
Yamaguchi University NUC
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Hiroshima University NUC
Nagasaki University NUC
Yamaguchi University NUC
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/16—Amides, e.g. hydroxamic acids
    • A61K31/18—Sulfonamides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
    • A61K31/4035—Isoindoles, e.g. phthalimide
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/425—Thiazoles
    • A61K31/426—1,3-Thiazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/12—Drugs for disorders of the urinary system of the kidneys

Definitions

  • the present invention relates to a preventive or therapeutic agent for polycystic kidney disease.
  • Autosomal dominant polycystic kidney disease is a genetic disease caused by a mutation in the polycystin complex (polycystin 1 and polycystin 2 form a tetramer in a ratio of 3:1), which is a mechanosensor calcium ion channel localized in the primary cilia of renal epithelial cells.
  • This autosomal dominant polycystic kidney disease is designated as an intractable disease (designated intractable disease 67), and the number of patients in Japan as of 1994 was estimated to be 31,000. By the age of 70, half of the patients will develop end-stage renal failure and will develop liver cysts and cerebral aneurysms.
  • octreotide a somatostatin analogue
  • mTOR inhibitors mimmalian target of rapamycin
  • venglastat a glucosylceramide synthase inhibitor
  • the present inventors have been conducting research into the promotion of cholesterol transport to the primary cilium, and have disclosed that oxysterol-binding protein (OSBP)-related proteins-3 (ORP3), which is localized in the primary cilium pocket, is responsible for cholesterol transport from peroxisomes to the primary cilium (see Non-Patent Document 3).
  • OSBP oxysterol-binding protein
  • ORP3 oxysterol-binding protein-related proteins-3
  • the objective of the present invention is to provide a new agent for preventing or treating polycystic kidney disease.
  • renal cilia sense the urine flow rate, and the thickness (diameter) of the renal tubule is regulated by a complex of polycystin 1, which is involved in the proliferation and differentiation of renal tubule cells and the sensing of urine flow on the renal cilia, and polycystin 2, which is a calcium channel.
  • polycystin 1 which is involved in the proliferation and differentiation of renal tubule cells and the sensing of urine flow on the renal cilia
  • polycystin 2 which is a calcium channel.
  • the present inventors previously found that when organelle adhesion between primary cilium and peroxisomes is induced by the FRB-FKBP system (rapamycin-dependent heteromolecular adhesion system), cholesterol in primary cilium increases in an ORP3/vesicle-associated membrane protein-associated protein (VAP-A) system-dependent manner. Therefore, we hypothesized that in patients with polycystic kidney disease, in which transport of polycystin complexes to primary cilium is impaired, promoting cholesterol transport to primary cilium by activating the ORP3/VAP-A system would improve the localization of polycystin complexes to primary cilium, and thus the progression of the disease would be suppressed by the recovery of renal function.
  • FRB-FKBP rapamycin-dependent heteromolecular adhesion system
  • a preventive or therapeutic agent for polycystic kidney disease comprising as an active ingredient a compound or a pharmacologic acceptable salt thereof that promotes intermolecular interaction between oxysterol-binding protein-related protein-3 (ORP3) and endoplasmic reticulum protein-related protein-A (VAP-A) in cells and increases cholesterol in the primary cilia of cells.
  • ORP3 oxysterol-binding protein-related protein-3
  • VAP-A endoplasmic reticulum protein-related protein-A
  • R 3A and R 4A are the same or different, Hydrogen atom, Halogen atoms, Nitro group, an optionally substituted C1-6 alkyl group, an optionally substituted C1-6 alkoxy group, or The group represents an optionally substituted phenyl group.
  • R 5A , R 6A , R 7A , R 8A and R 9A are the same or different; Hydrogen atoms, Halogen atoms, Nitro group, an optionally substituted C1-6 alkyl group, or It represents a C1-6 alkoxy group which may have a substituent.
  • the preventive or therapeutic agent for polycystic kidney disease which is a 4-amino-1-phenylpyrazolopyrimidine compound represented by the following formula (I): [4]
  • the compound having the effect of promoting the intermolecular interaction and increasing cholesterol in the primary cilium of the cell A compound represented by the following formula (I-1) (4-methoxypropylamino-1-(2,4-dimethylphenyl)-1H-pyrazolo[3,4-d]pyrimidine), A compound represented by formula (I-2) (4-(4-fluorobenzylamino)-1-(3-methylphenyl)-1H-pyrazolo[3,4-d]pyrimidine), A compound represented by formula (I-3) (4-(3,5-dimethylphenyl)amino-1-(4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidine), A compound represented by formula (I-4) (2-(4-ch
  • R 1B , R 2B , R 3B , R 4B , R 5B , R 7B , R 8B , R 9B and R 10B are the same or different and are Hydrogen atoms, Halogen atoms, Nitro group, an optionally substituted C1-6 alkyl group, an optionally substituted C1-6 alkoxy group, or The group represents an optionally substituted phenyl group.
  • R6B is A hydrogen atom, or It represents a C1-6 alkyl group which may have a substituent.
  • R 11B and R 12B are the same or different; Hydrogen atoms, an optionally substituted C1-6 alkyl group, a C1-6 alkoxy group which may be substituted; an optionally substituted C3-8 cycloalkyl group, an optionally substituted C3-6 cycloalkoxy group, or It represents a phenyl group which may have a substituent.
  • the preventive or therapeutic agent for polycystic kidney disease according to the above-mentioned [1], which is a 5-(sulfamoyl)benzamide compound represented by the following formula: [6]
  • the compound having the effect of promoting the intermolecular interaction and increasing cholesterol in the primary cilium of the cell A compound represented by the following formula (II-1): It is a compound represented by formula (II-2): The preventive or therapeutic agent for polycystic kidney disease described in [1] above.
  • R7C is Hydrogen atom, Halogen atoms, an optionally substituted C1-6 alkyl group, an optionally substituted C1-6 alkoxy group, or It represents a phenyl group which may have a substituent.
  • X represents an oxygen atom or CH2 .
  • the preventive or therapeutic agent for polycystic kidney disease according to the above-mentioned [1] which is a compound represented by the following formula (IV-1): [11]
  • the present invention makes it possible to provide a new agent for preventing or treating polycystic kidney disease that has a different mechanism of action from existing drugs.
  • FIG. 1 shows the results of measuring the luminescence of hit compounds obtained from a compound library using a screening platform constructed in Example 1 by introducing a split-luciferase system that targets the intermolecular interaction between ORP3 and VAPA.
  • FIG. 2A shows the results of observing primary cilia cholesterol using a confocal laser microscope when treated with compound A in the primary cilia cholesterol supply activity in Example 2.
  • Figure 2B shows the results of measuring the fluorescence intensity of Filipin III in primary cilia when treated with compound A in the primary cilia cholesterol supply activity in Example 2.
  • FIG. 3A shows the results of observing primary cilium cholesterol using a confocal laser microscope when treated with compounds A, C, and D in the primary cilium cholesterol supply activity in Example 2.
  • Figure 3B shows the results of measuring the fluorescence intensity of Filipin III in primary cilia when treated with compounds A, C, and D in the primary cilia cholesterol supply activity in Example 2.
  • FIG. 4A shows the results of observing primary cilia cholesterol using a confocal laser microscope when treated with compounds A, AB, AF, AL, AP, R, and V in the primary cilia cholesterol supply activity in Example 2.
  • Figure 4B shows the results of measuring the fluorescence intensity of Filipin III in primary cilia when treated with compounds A, AB, AF, AL, AP, R, and V in the primary cilia cholesterol supply activity in Example 2.
  • FIG. 4A shows the results of observing primary cilia cholesterol using a confocal laser microscope when treated with compounds A, AB, AF, AL, AP, R, and V in the primary cilia cholesterol supply activity in Example 2.
  • Figure 4B shows the results of measuring the fluorescence intensity of Filipin III in primary cilia when treated with compounds A, AB, AF
  • FIG. 5A shows the results of observing primary cilia cholesterol using a confocal laser microscope when treated with compounds A and I in the primary cilia cholesterol supply activity in Example 2.
  • Figure 5B shows the results of measuring the fluorescence intensity of Filipin III in primary cilia when treated with compounds A and I in the primary cilia cholesterol supply activity in Example 2.
  • FIG. 6A shows the results of confocal laser microscope observation of primary cilium cholesterol in the case of treatment with compound C in the recovery of primary cilium localization of polycystin-1 missense protein in Example 3.
  • FIG. 6B shows the results of measuring the fluorescence intensity of polycystin-1 in primary cilia when treated with compound C in the restoration of primary cilium localization of polycystin-1 missense protein in Example 3.
  • FIG. 7A shows the results of observing cysts formed from cells derived from the renal collecting duct of PC1 mutant mice treated with compounds A, C, and R using a confocal laser microscope after staining with Phalloidin (stains F-actin skeleton) and DAPI (stains DNA) in order to inhibit the increase in duct diameter in the cyst culture system in Example 4.
  • FIG. 7B shows the results of observing cysts formed from renal collecting duct-derived cells of PC2 mutant mice treated with compounds A, C, and R using a confocal laser microscope after staining with Phalloidin and DAPI in order to inhibit the increase in duct diameter in the cyst culture system in Example 4.
  • FIG. 7A shows the results of observing cysts formed from cells derived from the renal collecting duct of PC1 mutant mice treated with compounds A, C, and R using a confocal laser microscope after staining with Phalloidin and DAPI in order to inhibit the increase in duct diameter in the cyst culture system in Example 4.
  • FIG. 7A shows
  • FIG. 7C shows the results of determining the inner diameter/outer diameter ratio in merged images of cysts formed when treated with compounds A, C, and R in the inhibitory effect on the increase in ductal diameter in a cyst culture system in Example 4.
  • FIG. 8A shows the results of observing cysts formed from renal collecting duct-derived cells from PC2 mutant mice treated with compounds A, G, and R, and the existing drug tolvaptan, using a confocal laser microscope, after staining them with phalloidin and DAPI, in the inhibitory effect on the increase in ductal diameter in the cyst culture system in Example 5.
  • FIG. 8A shows the results of observing cysts formed from renal collecting duct-derived cells from PC2 mutant mice treated with compounds A, G, and R, and the existing drug tolvaptan, using a confocal laser microscope, after staining them with phalloidin and DAPI, in the inhibitory effect on the increase in ductal diameter in the cyst culture system in Example 5.
  • FIG. 8B shows the results of determining the inner diameter/outer diameter in merge images of cysts formed from renal collecting duct-derived cells from PC2 mutant mice when treated with compounds A, G, and R, and the existing drug tolvaptan, in the inhibitory effect on the increase in ductal diameter in a cyst culture system in Example 5.
  • FIG. 9A shows the results of observing cysts formed from cells derived from the renal collecting duct of PC2 mutant mice treated with compounds A, G, and compound G-OH, a derivative of compound G, using a confocal laser microscope after staining with phalloidin and DAPI in order to inhibit the increase in duct diameter in the cyst culture system in Example 5.
  • FIG. 9A shows the results of observing cysts formed from cells derived from the renal collecting duct of PC2 mutant mice treated with compounds A, G, and compound G-OH, a derivative of compound G, using a confocal laser microscope after staining with phalloidin and DAPI in order to inhibit the increase in duct diameter in the
  • FIG. 9B shows the results of determining the inner diameter/outer diameter in merge images of cysts formed from cells derived from the renal collecting duct of PC2 mutant mice treated with compounds A, G, and compound G-OH, a derivative of compound G, in the inhibitory effect on the increase in ductal diameter in a cyst culture system in Example 5.
  • FIG. 10A shows the results of observing cysts formed from renal collecting duct-derived cells from PC2 mutant mice treated with compounds A, G, B, Q, K, and J using a confocal laser microscope after staining with phalloidin and DAPI in order to inhibit the increase in duct diameter in the cyst culture system in Example 6.
  • FIG. 10A shows the results of observing cysts formed from renal collecting duct-derived cells from PC2 mutant mice treated with compounds A, G, B, Q, K, and J using a confocal laser microscope after staining with phalloidin and DAPI in order to inhibit the increase in duct diameter in the cyst culture system in Example 6.
  • FIG. 10B shows the results of determining the inner diameter/outer diameter in merged images of cysts formed from cells derived from the renal collecting duct of PC2 mutant mice treated with compounds A, G, B, Q, K, and J in the inhibitory effect on the increase in ductal diameter in a cyst culture system in Example 6.
  • FIG. 11A shows the results of observing cysts formed from renal collecting duct-derived cells from PC2 mutant mice treated with compounds A and I using a confocal laser microscope after staining with phalloidin and DAPI in order to inhibit the increase in duct diameter in a cyst culture system in Example 6.
  • FIG. 11A shows the results of observing cysts formed from renal collecting duct-derived cells from PC2 mutant mice treated with compounds A and I using a confocal laser microscope after staining with phalloidin and DAPI in order to inhibit the increase in duct diameter in a cyst culture system in Example 6.
  • FIG. 11B shows the results of determining the inner diameter/outer diameter in merged images of cysts formed from cells derived from the renal collecting duct of PC2 mutant mice treated with compounds A and I in the inhibitory effect on the increase in duct diameter in a cyst culture system in Example 6.
  • FIG. 12A shows the results of confocal laser microscope observation of polycystin-2 protein in the primary cilium of human induced pluripotent stem cells derived from a patient with polycystin-2 disease when treated with compound G-OH and the existing drug tolvaptan in the recovery of primary cilium localization of polycystin-2 protein in Example 7.
  • Figure B shows the results of measuring the amount of polycystin 2 protein in the primary cilium of human induced pluripotent stem cells derived from a patient with polycystin 2 kidney disease when treated with compound G-OH and the existing drug tolvaptan in the restoration of primary cilium localization of polycystin 2 protein (PC2) in Example 7.
  • FIG. 13A shows the results of observing PC1 in primary cilium using a confocal laser microscope when a feline kidney-derived epithelial cell line (CRFK cells) was treated with compound G, in relation to the increasing activity of polycystin-1 protein (PC1) in primary cilium in Example 8.
  • FIG. 13B shows the results of measuring the amount of polycystin-1 protein in the primary cilium of a feline kidney-derived epithelial cell line (CRFK cells) treated with compound G in relation to the increasing activity of polycystin-1 protein (PC1) in the primary cilium in Example 8.
  • the preventive or therapeutic agent for polycystic kidney disease of the present invention is not particularly limited as long as it is an agent for preventing or treating polycystic kidney disease that contains as an active ingredient a compound or a pharma- ceutically acceptable salt thereof that promotes intermolecular interactions between oxysterol-binding protein-related protein-3 (ORP3) and endoplasmic reticulum protein-related protein-A (VAP-A) in cells and increases cholesterol in the primary cilia of cells, and is hereinafter also referred to as "the preventive or therapeutic agent for polycystic kidney disease of this invention.”
  • ORP3 oxysterol-binding protein-related protein-3
  • VAP-A endoplasmic reticulum protein-related protein-A
  • polycystic kidney disease is not particularly limited, and examples thereof include autosomal dominant polycystic kidney disease (ADPKD) and autosomal recessive polycystic kidney disease (ARPKD), with autosomal dominant polycystic kidney disease (ADPKD) being preferred.
  • ADPKD autosomal dominant polycystic kidney disease
  • Autosomal dominant polycystic kidney disease (ADPKD) is caused by gene mutations in polycystic kidney and hepatic disease (PKD)1 on chromosome 16 and/or PKD2 on chromosome 4, which encode polycystin 1 protein (PC1) and polycystin 2 protein (PC2), respectively.
  • autosomal recessive polycystic kidney disease (ARPKD) is caused by a gene mutation in PKHD1 on chromosome 6.
  • the promotion of intermolecular interactions between oxysterol-binding protein-related protein-3 (ORP3) and endoplasmic reticulum protein-associated protein-A (VAP-A) in cells can be examined by known protein-protein interaction analysis methods, such as the split protein system and the GST-pull down method (Marion Weber-Boyvat et al., Experimental Cell Research Volume 331, Issue 2, 2011). e 2, 15 February 2015, Pages 278-291 (especially Figure 1); Wan Ting Saw et al., Methods Volume 90, November 2015, Pages 68-75; Stephanie Cabantous et al., Scientific Reports, volume 3, Article number: 2854 (2013); see Special Table 2003-503067).
  • ORP3 oxysterol-binding protein-related protein-3
  • VAP-A endoplasmic reticulum protein-associated protein-A
  • a fluorescently labeled protein is split so that it does not emit light by itself, and one of the split fragments of the fluorescently labeled protein is fused to ORP3 and the other to VAP-A.
  • a compound to be evaluated is added to cells expressing ORP3 and VAP-A fused with a fragment of the fluorescently labeled protein, and the cells are cultured.
  • the fluorescence intensity is then detected, and it can be evaluated by examining whether the fluorescence level increases when treated with the compound to be evaluated compared with the fluorescence level when not treated with the compound to be evaluated, or when a comparison compound or positive control compound is added.
  • Cells expressing ORP3 and VAP-A fused with a fragment of the fluorescently labeled protein can be adjusted by introducing an expression vector incorporating a polynucleotide encoding one of the fragments of the fluorescently labeled protein and a nucleotide encoding ORP3, and an expression vector incorporating a polynucleotide encoding the other fragment of the fluorescently labeled protein and a nucleotide encoding VAP-A into cells by a known method, and expressing them.
  • An example of a fluorescent label is green fluorescent protein (GFP).
  • evaluation can be performed by using an enzyme instead of the above-mentioned fluorescent label, adding a luminescent substrate as necessary, culturing, detecting the enzyme activity, and examining whether the enzyme activity increases when treated with the compound to be evaluated compared with the enzyme activity when the compound to be evaluated is not added or when a comparison compound is added.
  • enzymes include luciferases. Evaluation can also be performed by gradually changing the concentration of the compound to be evaluated and detecting the fluorescence intensity or enzyme activity in a concentration-dependent manner.
  • the compound to be evaluated When using a split protein system, the compound to be evaluated is added and the fluorescence intensity or enzyme activity is detected.
  • the fluorescence level or enzyme activity when the compound to be evaluated is not treated is set to 1. If the fluorescence level or enzyme activity reaches a predetermined value or more, the compound to be evaluated can be evaluated as having the ability to promote intermolecular interactions between oxysterol-binding protein-related protein-3 (ORP3) and endoplasmic reticulum protein-related protein-A (VAP-A) in cells.
  • ORP3 oxysterol-binding protein-related protein-3
  • VAP-A endoplasmic reticulum protein-related protein-A
  • the fluorescence level or enzyme activity of the treated cells is about 5, so if the relative level is 2.5 times or more, more preferably 2.8 times or more, the compound to be evaluated can be evaluated as having the ability to promote intermolecular interactions between oxysterol-binding protein-related protein-3 (ORP3) and endoplasmic reticulum protein-related protein-A (VAP-A) in cells.
  • ORP3 oxysterol-binding protein-related protein-3
  • VAP-A endoplasmic reticulum protein-related protein-A
  • the increase in cholesterol in the primary cilium of cells in the present specification can be achieved by known methods.
  • cells such as retinal pigment epithelial cells are treated with a compound to be evaluated for 4 hours at 37°C, stained with a cholesterol indicator such as Filipin III that emits fluorescence upon binding with cholesterol, or AcGFP1-D4 (a cholesterol probe in which a fluorescent protein is fused to a cholesterol-binding protein: see Miyamoto et al., Methods Mol Biol, 2022:doi: 10.1007/978-1-0716-1701-4_5.), and a primary cilium marker, and observed under a microscope to observe or detect cholesterol localized in the primary cilium.
  • a cholesterol indicator such as Filipin III that emits fluorescence upon binding with cholesterol
  • AcGFP1-D4 a cholesterol probe in which a fluorescent protein is fused to a cholesterol-binding protein: see Miyamoto et al., Methods Mol Biol, 2022:doi: 10.1007/978-1-0716
  • the compound to be evaluated is added to detect cholesterol localized in the primary cilium, and if the fluorescence level is 1.2 times or more, preferably 1.3 times or more, and more preferably 1.4 times or more in relative terms, relative to the level of 1 when the compound to be evaluated is not treated, the compound to be evaluated can be evaluated as having the effect of increasing cholesterol in the primary cilium of the cell.
  • the cholesterol-increasing effect in the primary cilium of the above-mentioned cells is ORP3/VAPA dependent. Whether or not the cholesterol-increasing effect in the primary cilium of the cells is ORP3/VAPA dependent can be evaluated by using wild-type retinal pigment epithelial cells as cells as well as mutant retinal pigment epithelial cells in which VAP-A or ORP3 is deleted or functionally mutated, and observing or detecting the activity of the compound to be evaluated to increase cholesterol in the primary cilium of the cells in the wild-type retinal pigment epithelial cells but not in the mutant retinal pigment epithelial cells.
  • the compound of the present invention that has the effect of promoting the intermolecular interaction between ORP3 and VAP-A in cells and increasing cholesterol in the primary cilium of cells preferably has the effect of restoring the localization of polycystin-1 to the primary cilium when mouse kidney collecting duct-derived cells (mIMCD3 cells: ATCC cat. no. CRL-2123) expressing polycystin-1 missense protein (a mutation in which cysteine at position 210 in the amino acid sequence of SEQ ID NO:1 is replaced with glycine) are used.
  • mIMCD3 cells ATCC cat. no. CRL-2123
  • polycystin-1 missense protein a mutation in which cysteine at position 210 in the amino acid sequence of SEQ ID NO:1 is replaced with glycine
  • the above-mentioned effect can be confirmed by treating the above-mentioned mutant mIMCD3 cells and, if necessary, wild-type mIMCD3 with the compound to be evaluated for 4 hours at 37°C, staining with polycystin-1 antibody and a primary cilium marker, and observing under a microscope to observe or detect the polycystin-1 protein localized to the primary cilium.
  • the compound of the present invention having the effect of promoting the intermolecular interaction between ORP3 and VAP-A in the cells and increasing cholesterol in the primary cilia of the cells preferably has an effect of suppressing the increase in tube diameter in a cyst culture system in which cyst formation by three-dimensional culture of renal tubular cells is reconstituted.
  • Whether or not the compound has an effect of suppressing the increase in tube diameter in a cyst culture system in which cyst formation by three-dimensional culture of renal tubular cells is reconstituted can be determined, for example, by three-dimensionally culturing mutant mIMCDC3 cells expressing polycystin 1 missense protein or mutant mIMCDC3 cells expressing polycystin 2 missense protein, mixing the compound to be evaluated and Matrigel, transferring to a chamber slide plate, and incubating the mixture at 37 ° C. and 5% CO 2 conditions until it solidifies. Then, a preheated medium is added to the solidified cell-Matrigel layer, and the mixture is incubated at 37 ° C.
  • the formed cyst is fixed, sliced, stained with Phalloidin and DAPI, and observed using a confocal laser microscope. Furthermore, the size of the epithelial lumen formed in the cyst in the merged image of the formed cyst is taken as the inner diameter, and the size of the entire cyst is measured as the outer diameter, and the inner diameter/outer diameter is calculated to evaluate the inhibitory effect on the increase in the duct diameter. Note that mutations in polycystin 1 protein or polycystin 2 protein reduce primary cilia activity, leading to cyst enlargement.
  • the compound of the present invention that promotes the intermolecular interaction between ORP3 and VAP-A in cells and has the effect of increasing cholesterol in the primary cilia of cells may have the effect of increasing cholesterol by directly acting on ORP3 or VAP-A, or may have the effect of increasing cholesterol by indirectly acting on ORP3 or VAP-A via a factor that can act on ORP3 or VAP-A.
  • a factor that can act on ORP3 or VAP-A examples include proteins, nucleic acids, and lipids.
  • the compound to be used as an active ingredient in the present agent for preventing or treating polycystic kidney disease is not particularly limited, and examples thereof include: A compound containing a 4-amino-1-phenylpyrazolopyrimidine skeleton (hereinafter, also referred to as a "4-amino-1-phenylpyrazolopyrimidine compound” or “compound (I)”). A compound containing a 5-(sulfamoyl)benzamide skeleton (hereinafter, also referred to as a “5-(sulfamoyl)benzamide compound” or “compound (II)”).
  • a compound containing an N-thiazolylbenzamide skeleton (hereinafter, also referred to as an “N-thiazolylbenzamide compound” or “compound (III)”)), examples include a compound containing an N-benzylsuccinimide skeleton (hereinafter, also referred to as an "N-benzylsuccinimide compound” or “compound (IV)”).
  • R 1A and R 2A are the same or different and Hydrogen atom, an optionally substituted C1-6 alkyl group, a C1-6 alkoxy group which may be substituted; a C2-6 alkenyl group optionally having a substituent, a C2-6 alkynyl group optionally having a substituent, an optionally substituted phenyl group, an aliphatic ring which may have a substituent, or The heterocycle may have a substituent.
  • R 3A and R 4A are the same or different, Hydrogen atom, Halogen atoms, Nitro group, an optionally substituted C1-6 alkyl group, an optionally substituted C1-6 alkoxy group, or The group represents an optionally substituted phenyl group.
  • R 5A , R 6A , R 7A , R 8A and R 9A are the same or different; Hydrogen atom, Halogen atoms, Nitro group, an optionally substituted C1-6 alkyl group, or It represents a C1-6 alkoxy group which may have a substituent.
  • C1-6 alkyl group there are no particular limitations on the C1-6 alkyl group as long as it is a linear or branched C1-6 alkyl group, and examples of such groups include linear or branched alkyl groups having 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, and isohexyl.
  • n- means normal
  • s- means secondary
  • t- means tertiary.
  • the C1-6 alkoxy group is a group represented by RO- (wherein R is an alkyl group), and there is no particular limitation on this "alkoxy group" as long as it is a linear or branched C1-6 alkoxy group, and examples thereof include linear or branched alkoxy groups having 1 to 6 carbon atoms, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, s-butoxy, t-butoxy, n-pentyloxy, isopentyloxy, neopentyloxy, n-hexyloxy, and isohexyloxy.
  • a C1-4 alkoxy group is preferable, and a methoxy group or a t-butoxy group is more preferable.
  • C2-6 alkenyl groups are not particularly limited, and examples include linear or branched alkenyl groups having 2 to 6 carbon atoms, such as vinyl, 1-propenyl, allyl, isopropenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-methyl-2-propenyl, and 1,3-butadienyl.
  • C2-6 alkynyl groups include, for example, linear or branched alkynyl groups having 2 to 6 carbon atoms, such as ethynyl, 1-propynyl, 2-propynyl, 1-methyl-2-propynyl, 1-butynyl, 2-butynyl, and 3-butynyl groups.
  • the "substituent" in the optionally substituted C1-6 alkyl group, optionally substituted C1-6 alkoxy group, optionally substituted C2-6 alkynyl group, optionally substituted C2-6 alkynyloxy group, optionally substituted phenyl group, optionally substituted aliphatic ring, optionally substituted heterocycle, optionally substituted C3-8 cycloalkyl group, and optionally substituted C3-8 cycloalkoxy group is not particularly limited, and examples thereof include, Halogen atoms, C1-6 alkyl group, C3-8 cycloalkyl group, C cycloalkenyl group, halo C1-6 alkyl group, C1-6 alkoxy group, an optionally substituted phenyl group or a nitro group; 1-acetylazetidin-3-ylmethyl group, C cycloalkylcarbonylamino group, Examples of the C1-6 alkyl group which may have a substituent at any position where it can be further
  • the C1-6 alkyl group which may have the above-mentioned substituent includes a halo C1-6 alkyl group
  • examples of the halo C1-6 alkyl group include a chloromethyl group, a chloroethyl group, a trifluoromethyl group, a 1,2-dichloro-n-propyl group, a 1-fluoro-n-butyl group, a perfluoro-n-pentyl group, and the like.
  • the C1-6 alkyl group which may have a substituent is not limited to these.
  • C3-8 cycloalkyl groups are not particularly limited, and examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups.
  • C3-8 cycloalkenyl groups include cyclopropenyl, cyclopentenyl, cyclohexenyl, and cyclooctenyl groups.
  • the double bond can be in any position.
  • Halogen atoms are not particularly limited, and examples include fluorine atoms, chlorine atoms, bromine atoms, and iodine atoms.
  • R 1A is an optionally substituted C1-6 alkyl group
  • a phenyl group which may have a substituent is preferable, a C1-6 alkyl group having a C1-6 alkoxy group; a C1-6 alkyl group having a phenyl group which may be substituted; a C1-6 alkyl group having a C3-6 cycloalkenyl group, and
  • An optionally substituted phenyl group is more preferred, a C1-6 alkyl group having a methoxy group or a hydroxy group, a C1-6 alkyl group having a phenyl group having a halogen atom at the 4-position, A C1-6 alkyl group having a cyclohexenyl group, and More preferably, the phenyl group has 1 to 3 C1-6 alkyl groups.
  • 4-methoxypropyl group, 4-fluorobenzyl group, 3,5-dimethylphenyl group, 4-chlorophenylethyl group, A (1-cyclohexenyl)ethyl group or a 4-hydroxy group is particularly preferred.
  • R 2A is preferably a hydrogen atom.
  • R 3A is preferably a hydrogen atom.
  • R 4A is preferably a hydrogen atom.
  • R 5A , R 6A , R 7A , R 8A and R 9A are each A hydrogen atom or a C1-6 alkyl group is preferred, A hydrogen atom or a methyl group is more preferred.
  • R 5A is a hydrogen atom
  • R 6A is a C1-6 alkyl group
  • R 7A is a hydrogen atom
  • R 8A is a hydrogen atom
  • R 9A is a hydrogen atom
  • R 5A is a C1-6 alkyl group
  • R 6A is a hydrogen atom
  • R 7A is a C1-6 alkyl group
  • R 8A is a hydrogen atom
  • R 9A is a hydrogen atom
  • R 6A is a hydrogen atom
  • R 7A is a C1-6 alkoxy group
  • R 8A is a hydrogen atom
  • R 9A is a hydrogen atom.
  • preferred compounds include the compound represented by the following formula (I-1) (4-methoxypropylamino-1-(2,4-dimethylphenyl)-1H-pyrazolo[3,4-d]pyrimidine), A compound represented by formula (I-2) (4-(4-fluorobenzylamino)-1-(3-methylphenyl)-1H-pyrazolo[3,4-d]pyrimidine), A compound represented by formula (I-3) (4-(3,5-dimethylphenyl)amino-1-(4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidine), A compound represented by formula (I-4) (2-(4-chlorophenyl)ethylamino-1-(3-methylphenyl)-1H-pyrazolo[3,4-d]pyrimidine), A compound represented by formula (I-5) (2-(1-methoxypropylamino-1-(2,4-dimethylphenyl)-1H-pyrazolo
  • 4-amino-1-phenylpyrazolopyrimidine compounds or salts thereof can be produced by known production methods.
  • the 4-amino-1-phenylpyrazolopyrimidine compound can be produced, for example, by using a known compound or a commercially available product as a raw material. A specific production method is shown in Scheme 1 described later.
  • R 1B , R 2B , R 3B , R 4B , R 5B , R 7B , R 8B , R 9B and R 10B are the same or different and each represents Hydrogen atoms, Halogen atoms, Nitro group, an optionally substituted C1-6 alkyl group, an optionally substituted C1-6 alkoxy group, or The group represents an optionally substituted phenyl group.
  • R6B is A hydrogen atom, or It represents a C1-6 alkyl group which may have a substituent.
  • R 11B and R 12B are the same or different; Hydrogen atoms, an optionally substituted C1-6 alkyl group, a C1-6 alkoxy group which may have a substituent; an optionally substituted C3-8 cycloalkyl group, an optionally substituted C3-6 cycloalkoxy group, or The group represents an optionally substituted phenyl group.
  • the C1-6 alkyl group, C1-6 alkoxy group, C3-8 cycloalkyl group, halogen atom, and substituents in general formula (II) in this specification are the same as the C1-6 alkyl group, C1-6 alkoxy group, halogen atom, and substituents in general formula (I) above.
  • C3-8 cycloalkoxy groups are not particularly limited, and examples include cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, cycloheptyloxy, and cyclooctyloxy.
  • R 1B , R 2B , R 3B , R 4B and R 5B may be a halogen atom, preferably a bromine atom, one to three, preferably two of R 1B , R 2B , R 3B , R 4B and R 5B may be a methyl group, and at least one, preferably both of R 2B and R 4B may be a methyl group.
  • R 11B is a cyclohexyl group or a methoxyphenylmethyl group.
  • R 1B , R 2B , R 3B , R 4B and R 5B are each A hydrogen atom, a halogen atom, or a C1-6 alkyl group is preferred, and a hydrogen atom, a bromine atom, or a methyl group is more preferred.
  • R 1B is a hydrogen atom
  • R 2B is a hydrogen atom
  • R 3B is a halogen atom or a C1-6 alkyl group
  • R 4B is a hydrogen atom
  • R 5B is a hydrogen atom
  • R 1B is a hydrogen atom
  • R 2B is a C1-6 alkyl group
  • R 3B is a hydrogen atom
  • R 4B is a C1-6 alkyl group
  • R 5B is a hydrogen atom
  • R 7B , R 8B , R 9B and R 10B are each A hydrogen atom, a halogen atom, or a C1-6 alkyl group is preferred; A hydrogen atom, a chlorine atom or a fluorine atom is more preferred.
  • R 7B is a halogen atom
  • R 8B is a hydrogen atom
  • R 9B is a halogen atom or a C1-6 alkyl group
  • R 10B is a hydrogen atom
  • R 7B is a chlorine atom or a fluorine atom
  • R 8B is a hydrogen atom
  • R 9B is a hydrogen atom
  • R 10B is a hydrogen atom
  • R 6B is preferably a hydrogen atom.
  • R 11B is preferably a hydrogen atom.
  • R 12B is preferably a C1-6 alkyl group which may have a substituent, or a C1-6 cycloalkyl group which may have a substituent, More preferred are C1-6 alkyl groups having a phenyl group having a C1-6 alkoxy group, and unsubstituted C1-6 cycloalkyl groups.
  • preferred compounds include the following compound (II-1): 2-chloro-N-(4-bromophenyl)-5-(cyclohexylaminosulfamoyl)benzamide, and compound (II-2): 2-fluoro-N-(3,5-dimethylphenyl)-5-[(1-methoxyphenylmethylamino)sulfamoyl)]benzamide.
  • 5-(sulfamoyl)benzamide compounds or salts thereof can be produced by known production methods.
  • the 5-(sulfamoyl)benzamide compound can be produced, for example, by using a known compound or a commercially available product as a raw material. A specific production method is shown in Scheme 2 or Scheme 3 described later.
  • N-thiazolylbenzamide compound represented by the general formula (III) The compound has the effect of promoting intermolecular interactions in the present prophylactic or therapeutic agent for polycystic kidney disease and increasing cholesterol in the primary cilia of cells
  • R 1C , R 2C , R 3C , R 4C , R 5C , R 8C , R 9C , R 10C , R 11C and R 12C are the same or different and each represents Hydrogen atoms, Halogen atoms, Nitro group, an optionally substituted C1-6 alkyl group, an optionally substituted C1-6 alkoxy group, or The group represents an optionally substituted phenyl group.
  • R6C is A hydrogen atom, or It represents a C1-6 alkyl group which may have a substituent.
  • R7C is Hydrogen atoms, Halogen atoms, an optionally substituted C1-6 alkyl group, an optionally substituted C1-6 alkoxy group, or The group represents an optionally substituted phenyl group.
  • the C1-6 alkyl group, C1-6 alkoxy group, halogen atom, and substituents in general formula (III) in this specification are the same as the C1-6 alkyl group, C1-6 alkoxy group, halogen atom, and substituents in general formula (I).
  • R 1C , R 2C , R 3C , R 4C , and R 5C in the formula is a nitro group, and it is more preferable that R 3C is a nitro group.
  • R 8C , R 9C , R 10C , R 11C , and R 12C in the formula is a methyl group, and it is more preferable that R 10C and/or R 11C are a methyl group.
  • R 1C , R 2C , R 4C , R 5C , R 7C , R 8C , R 9C , and R 12C in the formula are hydrogen atoms.
  • R 1C , R 2C , R 3C , R 4C , and R 5C are each A hydrogen atom, a halogen atom, a nitro group, or a C1-6 alkyl group is preferred; A hydrogen atom or a nitro group is more preferred.
  • R 1C is a hydrogen atom
  • R 2C is a hydrogen atom
  • R 3C is a hydrogen atom, a halogen atom, a nitro group, or a C1-6 alkyl group
  • R 4C is a hydrogen atom
  • R 5C is a hydrogen atom
  • R 1C is a hydrogen atom
  • R 2C is a C1-6 alkyl group
  • R 3C is a hydrogen atom
  • R 4C is a C1-6 alkyl group
  • R 5C is a hydrogen atom
  • R 6C is preferably a hydrogen atom or a C1-6 alkyl group; A hydrogen atom is more preferred.
  • R 7C is preferably a hydrogen atom.
  • R 8C , R 9C , R 10C , R 11C and R 12C are each A hydrogen atom or a C1-6 alkyl group is preferred, A hydrogen atom or a methyl group is more preferred.
  • R 8C is a hydrogen atom
  • R 9C is a hydrogen atom
  • R 10C is a hydrogen atom or a C1-6 alkyl group
  • R 11C is a hydrogen atom or a C1-6 alkyl group
  • R 12C is a hydrogen atom
  • R 8C is a hydrogen atom
  • R 9C is a hydrogen atom
  • R 10C is a C1-6 alkyl group
  • R 11C is a C1-6 alkyl group
  • R 12C is a hydrogen atom
  • N-thiazolylbenzamide compound represented by general formula (III) is the following compound (III-1): 4-nitro-N-[4-(2,3-dimethylphenyl)-(2-thiazolyl)benzamide.
  • N-thiazolylbenzamide compounds or salts thereof can be produced by known production methods.
  • the 5-(sulfamoyl)benzamide compound can be produced, for example, by using a known compound or a commercially available product as a raw material. A specific production method is shown in Scheme 4 described later.
  • N-benzylsuccinimide compound represented by the general formula (IV) The compound having the effect of promoting synthesis in the present prophylactic or therapeutic agent for polycystic kidney disease and the effect of increasing cholesterol in the primary cilia of cells is:
  • R 1D , R 2D , R 3D , R 4D and R 5D are the same or different and each represents A hydrogen atom, or It represents a C1-6 alkyl group which may have a substituent.
  • X represents an oxygen atom or CH2 .
  • the C1-6 alkyl group, C1-6 alkoxy group, halogen atom, and substituents in general formula (IV) in this specification are the same as the C1-6 alkyl group, C1-6 alkoxy group, halogen atom, and substituents in general formula (I).
  • R 1D , R 2D , R 3D , R 4D , and R 5D are each A hydrogen atom, a halogen atom, a nitro group, or a C1-6 alkyl group is preferred; A hydrogen atom is more preferred.
  • R 1D is a hydrogen atom
  • R 2D is a hydrogen atom
  • R 3D is a hydrogen atom, a halogen atom, a nitro group, or a C1-6 alkyl group
  • R 4D is a hydrogen atom
  • R 1D is a hydrogen atom
  • R 2C is a hydrogen atom
  • R 3C is a hydrogen atom
  • R 4C is a hydrogen atom
  • R 5C is a hydrogen atom.
  • X is preferably an oxygen atom or CH2 ; CH2 is more preferred.
  • N-benzylsuccinimide compound represented by general formula (IV) is the following compound (IV-1).
  • N-benzylsuccinamide compounds or salts thereof can be produced by known production methods.
  • the N-benzylsuccinimide compound can be produced, for example, by using a known compound or a commercially available product as a raw material. A specific production method is shown in Scheme 5 described later.
  • the salts in this specification are not particularly limited as long as they are medicamentously acceptable salts, and examples thereof include alkali metal salts (sodium salts, potassium salts, etc.), alkaline earth metal salts (calcium salts, magnesium salts, etc.), and salts with organic bases such as trimethylamine, triethylamine, pyridine, picoline, N-methylpyrrolidine, N-methylpiperidine, and N-methylmorpholine.
  • alkali metal salts sodium salts, potassium salts, etc.
  • alkaline earth metal salts calcium salts, magnesium salts, etc.
  • salts with organic bases such as trimethylamine, triethylamine, pyridine, picoline, N-methylpyrrolidine, N-methylpiperidine, and N-methylmorpholine.
  • examples thereof include acid addition salts of mineral acids such as hydrochlorides, hydrobromides, hydroiodides, sulfates, nitrates, and phosphates; and acid addition salts of organic acids such as benzoates, methanesulfonates, ethanesulfonates, benzenesulfonates, p-toluenesulfonates, maleates, fumarates, tartrates, citrates, and acetates.
  • mineral acids such as hydrochlorides, hydrobromides, hydroiodides, sulfates, nitrates, and phosphates
  • organic acids such as benzoates, methanesulfonates, ethanesulfonates, benzenesulfonates, p-toluenesulfonates, maleates, fumarates, tartrates, citrates, and acetates.
  • the compounds and salts thereof of the present invention may have one or more asymmetric centers in their structural formula, and may have two or more optical isomers and diastereomers, and the present invention encompasses all of the optical isomers and mixtures containing them in any ratio.
  • the compounds and salts thereof of the present invention may have two geometric isomers derived from a carbon-carbon double bond in their structural formula, and the present invention encompasses all of the geometric isomers and mixtures containing them in any ratio.
  • the compounds of the present invention and their salts may have multiple tautomers, and the present invention encompasses all of the tautomers and mixtures containing them in any ratio.
  • R 1A , R 2A , R 3A , R 4A , R 5A , R 6A , R 7A , R 8A , and R 9A are as defined above.
  • R 10A represents a halogen atom.
  • the manufacturing method (A-1) is a method for synthesizing a compound represented by general formula (I), and includes a step of reacting a compound represented by general formula (1A) with a compound represented by general formula (2A) in a solvent in the presence of a base.
  • the amount of the compound represented by general formula (1A) is not particularly limited as long as it is at least 1 equivalent relative to the compound represented by general formula (2A) and the desired reaction proceeds, but is usually at least 1 equivalent and no more than 200 equivalents.
  • bases used in this reaction include metal hydrides such as sodium hydride; organic lithiums such as methyllithium, butyllithium, sec-butyllithium, t-butyllithium, and hexyllithium; and metal amides such as lithium diisopropylamide, lithium hexamethyldisilazane, sodium hexamethyldisilazane, and potassium hexamethyldisilazane.
  • metal hydrides such as sodium hydride
  • organic lithiums such as methyllithium, butyllithium, sec-butyllithium, t-butyllithium, and hexyllithium
  • metal amides such as lithium diisopropylamide, lithium hexamethyldisilazane, sodium hexamethyldisilazane, and potassium hexamethyldisilazane.
  • the amount of base used in this reaction may be at least 1 equivalent relative to the compound represented by general formula (1A), and is not particularly limited as long as the desired reaction proceeds, but is preferably at least 1 equivalent and no more than 10 equivalents.
  • the solvent used in this reaction is not particularly limited as long as the desired reaction proceeds, but examples include benzene-based solvents such as benzene, toluene, xylene, mesitylene, chlorobenzene, and dichlorobenzene; and halogen-based solvents such as dichloromethane, dichloroethane, chloroform, and carbon tetrachloride. These solvents can be used alone or in a mixture of two or more in any ratio.
  • benzene-based solvents such as benzene, toluene, xylene, mesitylene, chlorobenzene, and dichlorobenzene
  • halogen-based solvents such as dichloromethane, dichloroethane, chloroform, and carbon tetrachloride.
  • the amount of solvent used in this reaction is not particularly limited as long as the desired reaction proceeds, but is usually 3 to 200 times by weight relative to the compound represented by general formula (1A).
  • the temperature at which this reaction is carried out is not particularly limited as long as the desired reaction proceeds, but is usually between 0°C and 150°C or below the boiling point of the solvent.
  • the time for carrying out this reaction is not particularly limited as long as the desired reaction proceeds, but is usually from 10 minutes to 24 hours.
  • a separation operation can be performed.
  • an aqueous solution any of the following can be used: an acidic aqueous solution in which hydrochloric acid, sulfuric acid, ammonium chloride, etc. are dissolved; an alkaline aqueous solution in which potassium hydroxide, sodium hydroxide, sodium carbonate, potassium carbonate, sodium hydrogen carbonate, potassium hydrogen carbonate, etc. are dissolved; saline solution, etc. can be used.
  • a solvent that is not compatible with water such as a benzene-based solvent such as toluene, xylene, benzene, chlorobenzene, or dichlorobenzene; an ester-based solvent such as ethyl acetate, isopropyl acetate, or butyl acetate; an ether-based solvent such as diethyl ether, diisopropyl ether, or methyl-t-butyl ether; a halogen-based solvent such as dichloromethane, dichloroethane, chloroform, or carbon tetrachloride; or a hydrocarbon-based solvent such as hexane, heptane, cyclohexane, or methylcyclohexane, as necessary.
  • solvents can be used alone or in any combination of two or more types in any ratio. There is no particular limit to the number of separations, and they can be performed according to the desired
  • reaction mixture containing the compound represented by general formula (I) obtained above can be dehydrated with a drying agent such as sodium sulfate or magnesium sulfate, but this is not essential.
  • reaction mixture containing the compound represented by general formula (I) obtained above can be subjected to solvent distillation under reduced pressure as long as the compound does not decompose.
  • reaction mixture containing the compound represented by general formula (I) obtained after distilling off the solvent can be purified by washing with an appropriate solvent, reprecipitation, recrystallization, column chromatography, etc.
  • the compound represented by general formula (II) of the present invention can be produced, for example, by the production method represented by the following scheme 2 (production method (B-1)) or the production method represented by the following scheme 3 (production method (B-2)), but the present invention is not limited thereto.
  • the manufacturing method (B-1) is a method for synthesizing a compound represented by general formula (II), and includes a step of reacting an amine compound represented by general formula (1B) with a compound represented by general formula (2B) in a solvent in the presence of a base.
  • the amount of the compound represented by general formula (1B) is not particularly limited as long as it is at least 1 equivalent relative to the compound represented by general formula (2B) and the desired reaction proceeds, but is usually at least 1 equivalent and no more than 200 equivalents.
  • bases used in this reaction include metal hydrides such as sodium hydride; organic lithiums such as methyllithium, butyllithium, sec-butyllithium, t-butyllithium, and hexyllithium; and metal amides such as lithium diisopropylamide, lithium hexamethyldisilazane, sodium hexamethyldisilazane, and potassium hexamethyldisilazane.
  • metal hydrides such as sodium hydride
  • organic lithiums such as methyllithium, butyllithium, sec-butyllithium, t-butyllithium, and hexyllithium
  • metal amides such as lithium diisopropylamide, lithium hexamethyldisilazane, sodium hexamethyldisilazane, and potassium hexamethyldisilazane.
  • the amount of base used in this reaction may be at least 1 equivalent relative to the compound represented by general formula (1B), and is not particularly limited as long as the desired reaction proceeds, but is preferably at least 1 equivalent and no more than 10 equivalents.
  • the solvent used in this reaction is not particularly limited as long as the desired reaction proceeds, but examples include benzene-based solvents such as benzene, toluene, xylene, mesitylene, chlorobenzene, and dichlorobenzene; and halogen-based solvents such as dichloromethane, dichloroethane, chloroform, and carbon tetrachloride. These solvents can be used alone or in a mixture of two or more in any ratio.
  • benzene-based solvents such as benzene, toluene, xylene, mesitylene, chlorobenzene, and dichlorobenzene
  • halogen-based solvents such as dichloromethane, dichloroethane, chloroform, and carbon tetrachloride.
  • the amount of solvent used in this reaction is not particularly limited as long as the desired reaction proceeds, but is usually 3 to 200 times by weight relative to the compound represented by general formula (1B).
  • the temperature at which this reaction is carried out is not particularly limited as long as the desired reaction proceeds, but is usually between 0°C and 150°C or below the boiling point of the solvent.
  • the time for carrying out this reaction is not particularly limited as long as the desired reaction proceeds, but is usually from 10 minutes to 24 hours.
  • a separation operation can be performed.
  • an aqueous solution any of the following can be used: an acidic aqueous solution in which hydrochloric acid, sulfuric acid, ammonium chloride, etc. are dissolved; an alkaline aqueous solution in which potassium hydroxide, sodium hydroxide, sodium carbonate, potassium carbonate, sodium hydrogen carbonate, potassium hydrogen carbonate, etc. are dissolved; saline solution, etc. can be used.
  • a solvent that is not compatible with water such as a benzene-based solvent such as toluene, xylene, benzene, chlorobenzene, or dichlorobenzene; an ester-based solvent such as ethyl acetate, isopropyl acetate, or butyl acetate; an ether-based solvent such as diethyl ether, diisopropyl ether, or methyl-t-butyl ether; a halogen-based solvent such as dichloromethane, dichloroethane, chloroform, or carbon tetrachloride; or a hydrocarbon-based solvent such as hexane, heptane, cyclohexane, or methylcyclohexane, as necessary.
  • solvents can be used alone or in any combination of two or more types in any ratio. There is no particular limit to the number of separations, and they can be performed according to the desired
  • reaction mixture containing the compound represented by general formula (II) obtained above can be dehydrated with a drying agent such as sodium sulfate or magnesium sulfate, but this is not essential.
  • reaction mixture containing the compound represented by general formula (II) obtained above can be subjected to solvent distillation under reduced pressure as long as the compound does not decompose.
  • reaction mixture containing the compound represented by general formula (II) obtained after distilling off the solvent can be purified by washing with an appropriate solvent, reprecipitation, recrystallization, column chromatography, etc.
  • the manufacturing method (B-2) is a method for synthesizing a compound represented by general formula (II), and includes a step of reacting a sulfonamide compound represented by general formula (3B) with an amine compound represented by general formula (4B) in a solvent in the presence of a base.
  • the amount of the compound represented by general formula (4B) is not particularly limited as long as it is at least 1 equivalent relative to the compound represented by general formula (3B) and the desired reaction proceeds, but is usually at least 1 equivalent and no more than 200 equivalents.
  • bases used in this reaction include metal hydrides such as sodium hydride; organic lithiums such as methyllithium, butyllithium, sec-butyllithium, t-butyllithium, and hexyllithium; and metal amides such as lithium diisopropylamide, lithium hexamethyldisilazane, sodium hexamethyldisilazane, and potassium hexamethyldisilazane.
  • metal hydrides such as sodium hydride
  • organic lithiums such as methyllithium, butyllithium, sec-butyllithium, t-butyllithium, and hexyllithium
  • metal amides such as lithium diisopropylamide, lithium hexamethyldisilazane, sodium hexamethyldisilazane, and potassium hexamethyldisilazane.
  • the amount of base used in this reaction may be at least 1 equivalent relative to the compound represented by general formula (4B), and is not particularly limited as long as the desired reaction proceeds, but is preferably at least 1 equivalent and no more than 10 equivalents.
  • the solvent used in this reaction is not particularly limited as long as the desired reaction proceeds, but examples include benzene-based solvents such as benzene, toluene, xylene, mesitylene, chlorobenzene, and dichlorobenzene; and halogen-based solvents such as dichloromethane, dichloroethane, chloroform, and carbon tetrachloride. These solvents can be used alone or in a mixture of two or more in any ratio.
  • benzene-based solvents such as benzene, toluene, xylene, mesitylene, chlorobenzene, and dichlorobenzene
  • halogen-based solvents such as dichloromethane, dichloroethane, chloroform, and carbon tetrachloride.
  • the amount of solvent used in this reaction is not particularly limited as long as the desired reaction proceeds, but is usually 3 to 200 times by weight relative to the compound represented by general formula (3B).
  • the temperature at which this reaction is carried out is not particularly limited as long as the desired reaction proceeds, but is usually between 0°C and 150°C or below the boiling point of the solvent.
  • the time for carrying out this reaction is not particularly limited as long as the desired reaction proceeds, but is usually from 10 minutes to 24 hours.
  • a separation operation can be performed.
  • an aqueous solution any of the following can be used: an acidic aqueous solution in which hydrochloric acid, sulfuric acid, ammonium chloride, etc. are dissolved; an alkaline aqueous solution in which potassium hydroxide, sodium hydroxide, sodium carbonate, potassium carbonate, sodium hydrogen carbonate, potassium hydrogen carbonate, etc. are dissolved; saline solution, etc. can be used.
  • a solvent that is not compatible with water such as a benzene-based solvent such as toluene, xylene, benzene, chlorobenzene, or dichlorobenzene; an ester-based solvent such as ethyl acetate, isopropyl acetate, or butyl acetate; an ether-based solvent such as diethyl ether, diisopropyl ether, or methyl-t-butyl ether; a halogen-based solvent such as dichloromethane, dichloroethane, chloroform, or carbon tetrachloride; or a hydrocarbon-based solvent such as hexane, heptane, cyclohexane, or methylcyclohexane, as necessary.
  • solvents can be used alone or in any combination of two or more types in any ratio. There is no particular limit to the number of separations, and they can be performed according to the desired
  • reaction mixture containing the compound represented by general formula (II) obtained above can be dehydrated with a drying agent such as sodium sulfate or magnesium sulfate, but this is not essential.
  • reaction mixture containing the compound represented by general formula (II) obtained above can be subjected to solvent distillation under reduced pressure as long as the compound does not decompose.
  • reaction mixture containing the compound represented by general formula (II) obtained after distilling off the solvent can be purified by washing with an appropriate solvent, reprecipitation, recrystallization, column chromatography, etc.
  • the compound of the present invention represented by the general formula (III) can be produced, for example, by a production method represented by the following scheme 4, but the present invention is not limited thereto.
  • Scheme 4> In the formula, R1C , R2C , R3C , R4C , R5C , R6C , R7C , R8C , R9C , R10C , R11C and R12C are as defined above.
  • R 13C represents a halogen atom.
  • the manufacturing method (C) is a method for synthesizing a compound represented by general formula (III), and includes a step of reacting a thiazole compound represented by general formula (1C) with a compound represented by general formula (2C) in a solvent in the presence of a base.
  • the amount of the compound represented by general formula (1C) is not particularly limited as long as it is at least 1 equivalent relative to the compound represented by general formula (2C) and the desired reaction proceeds, but is usually at least 1 equivalent and no more than 200 equivalents.
  • bases used in this reaction include metal hydrides such as sodium hydride; organic lithiums such as methyllithium, butyllithium, sec-butyllithium, t-butyllithium, and hexyllithium; and metal amides such as lithium diisopropylamide, lithium hexamethyldisilazane, sodium hexamethyldisilazane, and potassium hexamethyldisilazane.
  • metal hydrides such as sodium hydride
  • organic lithiums such as methyllithium, butyllithium, sec-butyllithium, t-butyllithium, and hexyllithium
  • metal amides such as lithium diisopropylamide, lithium hexamethyldisilazane, sodium hexamethyldisilazane, and potassium hexamethyldisilazane.
  • the amount of base used in this reaction may be at least 1 equivalent relative to the compound represented by general formula (1C), and is not particularly limited as long as the desired reaction proceeds, but is preferably at least 1 equivalent and no more than 10 equivalents.
  • the solvent used in this reaction is not particularly limited as long as the desired reaction proceeds, but examples include benzene-based solvents such as benzene, toluene, xylene, mesitylene, chlorobenzene, and dichlorobenzene; and halogen-based solvents such as dichloromethane, dichloroethane, chloroform, and carbon tetrachloride. These solvents can be used alone or in a mixture of two or more in any ratio.
  • benzene-based solvents such as benzene, toluene, xylene, mesitylene, chlorobenzene, and dichlorobenzene
  • halogen-based solvents such as dichloromethane, dichloroethane, chloroform, and carbon tetrachloride.
  • the amount of solvent used in this reaction is not particularly limited as long as the desired reaction proceeds, but is usually 3 to 200 times by weight relative to the compound represented by general formula (1C).
  • the temperature at which this reaction is carried out is not particularly limited as long as the desired reaction proceeds, but is usually between 0°C and 150°C or below the boiling point of the solvent.
  • the time for carrying out this reaction is not particularly limited as long as the desired reaction proceeds, but is usually from 10 minutes to 24 hours.
  • a separation operation can be performed.
  • an aqueous solution any of the following can be used: an acidic aqueous solution in which hydrochloric acid, sulfuric acid, ammonium chloride, etc. are dissolved; an alkaline aqueous solution in which potassium hydroxide, sodium hydroxide, sodium carbonate, potassium carbonate, sodium hydrogen carbonate, potassium hydrogen carbonate, etc. are dissolved; saline solution, etc. can be used.
  • a solvent that is not compatible with water such as a benzene-based solvent such as toluene, xylene, benzene, chlorobenzene, or dichlorobenzene; an ester-based solvent such as ethyl acetate, isopropyl acetate, or butyl acetate; an ether-based solvent such as diethyl ether, diisopropyl ether, or methyl-t-butyl ether; a halogen-based solvent such as dichloromethane, dichloroethane, chloroform, or carbon tetrachloride; or a hydrocarbon-based solvent such as hexane, heptane, cyclohexane, or methylcyclohexane, as necessary.
  • solvents can be used alone or in any combination of two or more types in any ratio. There is no particular limit to the number of separations, and they can be performed according to the desired
  • reaction mixture containing the compound represented by general formula (III) obtained above can be dehydrated with a drying agent such as sodium sulfate or magnesium sulfate, but this is not essential.
  • reaction mixture containing the compound represented by general formula (III) obtained above can be subjected to solvent distillation under reduced pressure as long as the compound does not decompose.
  • reaction mixture containing the compound represented by general formula (III) obtained after distilling off the solvent can be purified by washing with an appropriate solvent, reprecipitation, recrystallization, column chromatography, etc.
  • the compound represented by general formula (IV) of the present invention can be produced, for example, by the production method represented by the following scheme 5, but the present invention is not limited thereto.
  • the manufacturing method (D) is a method for synthesizing an N-benzylsuccinimide compound represented by general formula (IV), and includes a step of reacting a compound represented by general formula (1D) with an N-maleimide compound represented by general formula (2D) in the absence of a solvent or in a solvent.
  • the amount of the compound represented by general formula (1D) is not particularly limited as long as it is at least 1 equivalent relative to the compound represented by general formula (2D) and the desired reaction proceeds, but is usually at least 1 equivalent and no more than 200 equivalents.
  • the type of solvent is not particularly limited as long as the desired reaction proceeds, but examples include benzene-based solvents such as benzene, toluene, xylene, mesitylene, chlorobenzene, and dichlorobenzene; and halogen-based solvents such as dichloromethane, dichloroethane, chloroform, and carbon tetrachloride. These solvents can be used alone or in a mixture of two or more types in any ratio.
  • the amount of solvent used in this reaction is not particularly limited as long as the desired reaction proceeds, but is usually 3 to 200 times by weight relative to the compound represented by general formula (1D).
  • Additives such as catalysts can be added to this reaction if necessary.
  • the temperature at which this reaction is carried out is not particularly limited as long as the desired reaction proceeds, but is usually between -78°C and 250°C or below the boiling point of the solvent.
  • the time for carrying out this reaction is not particularly limited as long as the desired reaction proceeds, but is usually from 10 minutes to 24 hours.
  • a separation operation can be performed.
  • an aqueous solution any of the following can be used: an acidic aqueous solution in which hydrochloric acid, sulfuric acid, ammonium chloride, etc. are dissolved; an alkaline aqueous solution in which potassium hydroxide, sodium hydroxide, sodium carbonate, potassium carbonate, sodium hydrogen carbonate, potassium hydrogen carbonate, etc. are dissolved; saline solution, etc. can be used.
  • a solvent that is not compatible with water such as a benzene-based solvent such as toluene, xylene, benzene, chlorobenzene, or dichlorobenzene; an ester-based solvent such as ethyl acetate, isopropyl acetate, or butyl acetate; an ether-based solvent such as diethyl ether, diisopropyl ether, or methyl-t-butyl ether; a halogen-based solvent such as dichloromethane, dichloroethane, chloroform, or carbon tetrachloride; or a hydrocarbon-based solvent such as hexane, heptane, cyclohexane, or methylcyclohexane, as necessary.
  • solvents can be used alone or in any combination of two or more types in any ratio. There is no particular limit to the number of separations, and they can be performed according to the desired
  • reaction mixture containing the compound represented by general formula (IV) obtained above can be dehydrated with a drying agent such as sodium sulfate or magnesium sulfate, but this is not essential.
  • reaction mixture containing the compound represented by general formula (IV) obtained above can be subjected to solvent distillation under reduced pressure as long as the compound does not decompose.
  • reaction mixture containing the compound represented by general formula (IV) obtained after distilling off the solvent can be purified by washing with an appropriate solvent, reprecipitation, recrystallization, column chromatography, etc.
  • the subjects for administration of the present agent for preventing or treating polycystic kidney disease include mammals, specifically humans, cats, dogs, monkeys, cows, horses, mice, rats, hamsters, guinea pigs, rabbits, goats, pigs, and sheep.
  • Subjects to which the present preventive or therapeutic agent for polycystic kidney disease is administered include subjects who have been diagnosed with polycystic kidney disease or subjects who have been diagnosed as needing prevention of polycystic kidney disease.
  • the agent can also be used as a preventive or therapeutic agent for autosomal dominant polycystic kidney disease to be administered to subjects with gene mutations in PKD1 and/or PKD2.
  • the present agent for preventing or treating polycystic kidney disease may contain pharma- ceutical acceptable additives.
  • the additives include saline, buffered saline, dextrose, water for injection, glycerol, ethanol, and combinations thereof, stabilizers, solubilizers and surfactants, buffers and preservatives, isotonicity agents, bulking agents, and lubricants.
  • the dosage of the present agent for preventing or treating polycystic kidney disease to a subject varies depending on age, body weight, symptoms, therapeutic effect, administration method, treatment time, etc., but typically, per adult, the content of the compound as the active ingredient per dose is in the range of 0.1 mg to 1000 mg, preferably 1 mg to 500 mg, and can be administered independently four times, three times, two times or once a day, every other day, every second day, every third day, every fourth day, every fifth day, every seventh day, every eighth day, every ninth day, twice a week, once a month, or twice a month.
  • the method of administration is not particularly limited, but examples include oral administration, intravenous administration, intramuscular administration, subcutaneous administration, transdermal administration, and nasal administration.
  • the agent for preventing or treating polycystic kidney disease may be used in combination with other drugs for kidney diseases, as well as with antihypertensive agents, lipid-lowering agents, hypoglycemic agents, etc.
  • the inventors introduced a split-luciferase system that targets the intermolecular interaction between ORP and VAP-A to construct a screening platform, and searched for compounds that promote the intermolecular interaction between ORP3 and VAPA. Specifically, first, a polynucleotide encoding ORP3 or VAPA was incorporated into an expression vector attached to the NanoBiT (registered trademark) Protein: Protein Interaction System (Promega), and then introduced into HEK293T cells by lipofection.
  • NanoBiT registered trademark
  • Protein Protein Interaction System
  • split-luciferase stably expressing HEK293T cell clone split-luciferase constitutively expressing HEK293T cells that detect the interaction between ORP3 and VAPA
  • HEK293 cells stably expressing split luciferase obtained above or parent HEK293 cells were seeded on a 96-well plate.
  • the compounds of the library and Nano-Glo Live Cell Reagent (Promega), a luminescent substrate for split-luciferase, were added and cultured, and immediately after treatment, the plate was placed in a luminescence image acquisition device (LAS-4000: GE Healthcare) and images were continuously acquired up to 4 hours after the addition of the substrate. The acquired images were analyzed using the Multi gauge software attached to the LAS-4000, and luminescence quantitative data for each well was obtained.
  • the split-luciferase expression system described above was created by fusing a 17.6 kDa Large BiT (LgBiT) to ORP3 and a Small BiT (SmBiT) consisting of 11 amino acids to VAP-A, resulting in a split-luciferase expression system in which luciferase is formed by the interaction between ORP3 and VAP-A.
  • LgBiT Large BiT
  • SmBiT Small BiT
  • Example 2 Increase in cholesterol in primary cilia
  • 10 compounds (compounds A, C, D, I, R, V, AB, AF, AL, and AP) were evaluated for the increase in cholesterol in primary cilia.
  • hTERT-immortalized human retinal pigment epithelial cells (hTERT-RPE1 cells: ATCC CRL-4000) were treated with the hit compounds for 4 hours at 37°C, and then multi-staining was performed with the cholesterol indicator Filipin III (Merck), anti-acetylated-tubulin antibody (Merck) which is a primary cilium marker, and anti-Pericentrin antibody (Bethyl) which is a centrosome marker.
  • Figures 3A and B show that in hTERT-RPE1 cells, treatment with compounds A and C increased cholesterol in the primary cilia in a dose-dependent manner, as revealed by Filipin III and immunostaining. On the other hand, compound D did not show any activity in increasing cholesterol in the primary cilia.
  • Figures 4A and B show that the cholesterol in the primary cilia in hTERT-RPE1 cells was increased in a dose-dependent manner by treatment with compounds A, R, and V, as revealed by Filipin III and immunostaining.
  • Compounds AB, AF, AL, and AP did not show any activity in increasing cholesterol in the primary cilia.
  • Figures 5A and B show that cholesterol in the primary cilia of hTERT-RPE1 cells is increased by treatment with compound I, as revealed by Filipin III and immunostaining.
  • Compounds A, C, I, and R are shown below.
  • Compound A is the above compound (III-1)
  • compound C is the above compound (IV-1)
  • compound I is the above compound (II-2)
  • compound R is the above compound (I-5).
  • Example 3 Restoration of Localization of Polycystin-1 Missense Protein to Primary Cilia Using mouse kidney collecting duct-derived cells (mIMCD3 cells: ATCC cat. no. CRL-2123) expressing polycystin-1 missense protein (a mutation in which cysteine at position 210 in the amino acid sequence of SEQ ID NO:1 is replaced with glycine), it was investigated whether Compound C has an effect of restoring the localization of polycystin-1 to primary cilia.
  • mIMCD3 cells ATCC cat. no. CRL-2123
  • polycystin-1 missense protein a mutation in which cysteine at position 210 in the amino acid sequence of SEQ ID NO:1 is replaced with glycine
  • the above mutant mIMCD3 cells were treated with compound C for 4 hours at 37°C, and then multi-stained with polycystin 1 antibody (Santa Cruz), anti-acetylated-tubulin antibody (Merck), a primary cilium marker, and anti-FGFR10P antibody (Proteintech), a centrosome marker.
  • polycystin 1 antibody Santa Cruz
  • Anti-acetylated-tubulin antibody Merck
  • a primary cilium marker
  • anti-FGFR10P antibody Proteintech
  • Figures 6A and 6B show the results.
  • Non-mutated mIMCD3 cells wild type were used as a control.
  • Figures 6A and B confirm that compound C has the effect of restoring the localization of polycystin-1 to the primary cilium, even when the polycystin-1 protein is mutated.
  • Example 4 Inhibitory effect on duct diameter increase in a cyst culture system reconstituting cyst formation by 3D culture of renal tubular cells - Compounds A, C, R, and V - We used mutant mIMCDC3 cells expressing polycystin 1 missense protein or mutant mIMCDC3 cells expressing polycystin 2 missense protein to examine whether each compound has the effect of normalizing cyst size. Mutations in polycystin 1 protein or polycystin 2 protein reduce primary cilia activity, leading to cyst enlargement.
  • PC1 mutant mouse kidney collecting duct-derived cells PC1 mutant mIMCD3 expressing polycystin 1 missense protein (mutation in which cysteine at position 210 of the amino acid sequence in SEQ ID NO: 1 is replaced with glycine) or PC2 mutant mIMCD3 cells expressing polycystin 2 missense protein (mutation in which tryptophan at position 412 of the amino acid sequence in SEQ ID NO: 2 (corresponding to position 414 of human polycystin 2 missense protein) is replaced with glycine) were prepared in mIMCD3 cells (ATCC cat.
  • Each mutant mIMCDC3 cell was suspended in DMEM/F-12 (L-glutamine) medium supplemented with 10% (vol/vol) FBS and 1% (vol/vol) pen-strep, and compound A, C, R, or V at 20 ⁇ M or cholesterol at 200 ⁇ M was mixed with matrigel and transferred to an 8-well chamber slide plate. The mixture was incubated (15 to 30 minutes) at 37°C under 5% CO2 conditions until solidification. Then, pre-heated medium was added to the solidified cell-matrigel layer, and the mixture was incubated at 37°C under 5% CO2 conditions while exchanging the medium until cyst formation.
  • the formed cysts were fixed and sliced. Furthermore, F-actin was stained with phalloidin and DNA with DAPI, and then the slices were photographed using a fluorescent microscope. Furthermore, the size of the epithelial lumen formed within the cyst in the merged image of the formed cyst was measured as the inner diameter, and the size of the entire cyst was measured as the outer diameter, and the inner diameter/outer diameter was calculated. The results are shown in Figures 7A to C. In Figure 7A, the dotted line in the merged image indicates the width of the inner diameter.
  • Example 5 Inhibitory effect on duct diameter increase in a cyst culture system reconstituting cyst formation by 3D culture of renal tubular cells - Comparison with tolvaptan - Using the above PC2 mutant mIMCDC3 cells, compounds A, G, and R, and an existing drug, tolvaptan, were added at 3.75 ⁇ M, 7.5 ⁇ M, and 15 ⁇ M, respectively, to examine whether each compound has an inhibitory effect on the increase in cystic duct diameter in the same manner as in Example 4. The results are shown in Figures 8A and 8B.
  • FIGS 9A and B confirm that not only compound G but also its derivative compound G-OH has the effect of improving cyst size.
  • Compounds G and G-OH are shown below.
  • Compound G is compound (I-1) above, and compound G-OH is compound (I-6) above.
  • Example 6 Inhibitory effect on duct diameter increase in a cyst culture system reconstituting cyst formation by 3D culture of renal tubular cells - Compounds B, Q, K, J, and I -
  • compounds A, C, and R were used, and compounds B, Q, K, J, and I were also used to similarly examine the effect of suppressing the increase in tube diameter.
  • the above-mentioned PC2 mutant mIMCDC3 cells were used as cells, and compounds A, G, B, Q, K, J, and I were used as compounds.
  • Compounds B, Q, K, and J are shown below.
  • Compound B is the above compound (II-1)
  • compound Q is the above compound (I-4)
  • compound K is the above compound (I-3)
  • compound J is the above compound (I-2).
  • Example 7 Restoration effect of polycystin 2 protein (PC2) on primary cilia localization in human induced pluripotent stem cells derived from patients with polycystic kidney disease - Comparison with tolvaptan Using induced pluripotent stem cells derived from patients with autosomal dominant polycystic kidney disease (HPS2192 cells: RIKEN BRC RRID: CVCL_YS47 and HPS2636 cells: RIKEN BRC RRID: CVCL_YS58: RIKEN BioResource Research Center), the compound G-OH was added at 5 ⁇ M, 10 ⁇ M, or 20 ⁇ M, or the existing drug tolvaptan was added at 20 ⁇ M to investigate whether it has an effect of restoring the localization of polycystin-2 protein to primary cilia.
  • PC2192 cells RIKEN BRC RRID: CVCL_YS47
  • HPS2636 cells RIKEN BRC RRID: CVCL_YS58: RIK
  • compound G-OH has the effect of restoring the localization of polycystin 2 to the primary cilium in human induced pluripotent stem cells derived from patients with polycystin 2, even when polycystin 2 is mutated.
  • concentration dependency of the above-mentioned restorative effect of compound G-OH was examined, immunostaining revealed that polycystin 2 in the primary cilium increased in a dose-dependent manner.
  • the existing drug tolvaptan did not show any activity to increase the amount of polycystin 2 in the primary cilium.
  • Example 8 Effect of Increasing Primary Cilia Localization of Polycystin 1 Protein (PC1) in Feline Cells
  • PC1 in primary cilia of compound G was evaluated.
  • CRFK cells an epithelial cell line derived from feline kidney, were prepared and treated with compound G (final concentration 20 ⁇ M) for 8, 16, or 24 hours at 37° C., and then multi-stained with anti-polycystin 1 antibody (Santa Cruz), anti-acetylated-tubulin antibody (Merck), which is a primary cilium marker, and anti-FGFR1OP antibody (Proteintech), which is a centrosome marker, and PC1 in primary cilia was observed using a confocal laser microscope to measure the fluorescence intensity of PC1 in primary cilia. The results are shown in FIGS. 13A and B.
  • compound G has the effect of increasing PC1 in primary cilia in CRFK cells, an epithelial cell line derived from feline kidney.

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Abstract

La présente invention aborde le problème de la fourniture d'un nouvel agent prophylactique ou thérapeutique contre des maladies rénales. L'invention concerne un agent prophylactique ou thérapeutique contre une maladie rénale polykystique, l'agent comprenant, en tant que principe actif, un composé qui favorise l'interaction intermoléculaire entre une protéine 3 liée à la protéine de liaison à l'oxystérol (ORP3) et une protéine associée à la protéine vésiculaire endoplasmique A (VAP-A) dans une cellule, et présente un effet d'augmentation du cholestérol dans les cils primaires de la cellule, ou un sel pharmaceutiquement acceptable de celui-ci.
PCT/JP2024/042728 2023-12-04 2024-12-03 Agent prophylactique ou thérapeutique contre la polykystose rénale Pending WO2025121320A1 (fr)

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Citations (5)

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JP2011503103A (ja) * 2007-11-07 2011-01-27 フォールドアールエックス ファーマシューティカルズ インコーポレーティッド タンパク質輸送の調節方法
JP2011517674A (ja) * 2008-03-25 2011-06-16 ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア 嚢胞性線維症膜コンダクタンス制御因子の水溶性小分子阻害剤
JP2013511541A (ja) * 2009-11-18 2013-04-04 プレキシコン インコーポレーテッド キナーゼ調節のための化合物および方法、ならびにその適応
JP2022529814A (ja) * 2019-04-24 2022-06-24 ガラパゴス・ナムローゼ・フェンノートシャップ 腎臓病の治療のための新規化合物及びその医薬組成物
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JP2011503103A (ja) * 2007-11-07 2011-01-27 フォールドアールエックス ファーマシューティカルズ インコーポレーティッド タンパク質輸送の調節方法
JP2011517674A (ja) * 2008-03-25 2011-06-16 ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア 嚢胞性線維症膜コンダクタンス制御因子の水溶性小分子阻害剤
JP2013511541A (ja) * 2009-11-18 2013-04-04 プレキシコン インコーポレーテッド キナーゼ調節のための化合物および方法、ならびにその適応
JP2022529814A (ja) * 2019-04-24 2022-06-24 ガラパゴス・ナムローゼ・フェンノートシャップ 腎臓病の治療のための新規化合物及びその医薬組成物
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