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WO2021221444A1 - Ubr 박스 도메인 리간드로의 화합물 - Google Patents

Ubr 박스 도메인 리간드로의 화합물 Download PDF

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Publication number
WO2021221444A1
WO2021221444A1 PCT/KR2021/005335 KR2021005335W WO2021221444A1 WO 2021221444 A1 WO2021221444 A1 WO 2021221444A1 KR 2021005335 W KR2021005335 W KR 2021005335W WO 2021221444 A1 WO2021221444 A1 WO 2021221444A1
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Prior art keywords
ubr
compound
protein
box domain
domain
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PCT/KR2021/005335
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English (en)
French (fr)
Inventor
권용태
김현태
나정은
서유진
지창훈
최하림
이지은
허아정
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Autotac Inc
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Autotac Inc
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Priority to EP21797495.5A priority Critical patent/EP4144720B1/en
Priority to JP2022563901A priority patent/JP7626347B2/ja
Priority to CN202180044509.4A priority patent/CN115768744B/zh
Priority to KR1020227037601A priority patent/KR102891700B1/ko
Priority to US17/921,688 priority patent/US12509420B2/en
Publication of WO2021221444A1 publication Critical patent/WO2021221444A1/ko
Anticipated expiration legal-status Critical
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    • C07—ORGANIC CHEMISTRY
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    • C07C311/48—Amides of sulfonic acids, i.e. compounds having singly-bound oxygen atoms of sulfo groups replaced by nitrogen atoms, not being part of nitro or nitroso groups having nitrogen atoms of sulfonamide groups further bound to another hetero atom
    • C07C311/49—Amides of sulfonic acids, i.e. compounds having singly-bound oxygen atoms of sulfo groups replaced by nitrogen atoms, not being part of nitro or nitroso groups having nitrogen atoms of sulfonamide groups further bound to another hetero atom to nitrogen atoms
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    • A61K31/16—Amides, e.g. hydroxamic acids
    • A61K31/165—Amides, e.g. hydroxamic acids having aromatic rings, e.g. colchicine, atenolol, progabide
    • A61K31/166—Amides, e.g. hydroxamic acids having aromatic rings, e.g. colchicine, atenolol, progabide having the carbon of a carboxamide group directly attached to the aromatic ring, e.g. procainamide, procarbazine, metoclopramide, labetalol
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    • A61K31/167—Amides, e.g. hydroxamic acids having aromatic rings, e.g. colchicine, atenolol, progabide having the nitrogen of a carboxamide group directly attached to the aromatic ring, e.g. lidocaine, paracetamol
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    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
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    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/451—Non condensed piperidines, e.g. piperocaine having a carbocyclic group directly attached to the heterocyclic ring, e.g. glutethimide, meperidine, loperamide, phencyclidine, piminodine
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    • C07D211/10—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms with radicals containing only carbon and hydrogen atoms attached to ring carbon atoms
    • C07D211/14—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms with radicals containing only carbon and hydrogen atoms attached to ring carbon atoms with hydrocarbon or substituted hydrocarbon radicals attached to the ring nitrogen atom
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    • C07D295/12—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly or doubly bound nitrogen atoms
    • C07D295/135—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly or doubly bound nitrogen atoms with the ring nitrogen atoms and the substituent nitrogen atoms separated by carbocyclic rings or by carbon chains interrupted by carbocyclic rings
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    • C07D295/16—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms
    • C07D295/20—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms by radicals derived from carbonic acid, or sulfur or nitrogen analogues thereof
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    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
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    • C07C2602/04—One of the condensed rings being a six-membered aromatic ring
    • C07C2602/08—One of the condensed rings being a six-membered aromatic ring the other ring being five-membered, e.g. indane

Definitions

  • the disclosure herein relates to compounds as UBR box domain ligands.
  • the UBR box domain is a domain commonly present in UBR (Ubiquitin protein ligase E3 component n-recognin) protein of the N-end rule pathway.
  • UBR box domain is known as a domain to which a substrate binds. It is known that the UBR box domain is essential for forming a multi-ubiquitin chain on the substrate by binding to the N-terminal residue of the substrate, and the substrate is degraded through this process.
  • the present specification relates to a compound that acts as a ligand that binds to the UBR box domain.
  • the protein in vivo may be degraded depending on the sequence of the N-terminal residue, and this degradation pathway is known as the N-end rule pathway. That is, the N-terminal pathway law is a protein degradation system that uses the N-terminus of a specific protein as a degradation signal.
  • the N-terminal pathway rule may include the following proteolytic process.
  • N-recognin recognizes an N-terminal degradation signal of a protein, and the N-recognin can degrade the protein by defective ubiquitin in the protein to be degraded.
  • the N-terminal degradation signal is a residue having a positive charge at the N-terminus (type 1; eg, arginine, lysine, histidine) or a large hydrophobic residue (type 2; phenylalanine, leucine, tryptophan, isoleucine, tyrosine) having a may include
  • type 1 eg, arginine, lysine, histidine
  • type 2 phenylalanine, leucine, tryptophan, isoleucine, tyrosine
  • the present inventors first discovered or cloned N-recognins UBR1, UBR2, UBR3, and UBR5, and revealed that they have a UBR box domain as a substrate recognition domain (Tasaki et al.
  • the ubiquitinated substrate made by binding N-recognin with the N-end rule ligand is delivered to the proteasome and degraded into short peptides.
  • certain N-terminal residues Nt-Arg, Nt-His, Nt-Lys, Nt-Trp, Nt-Phe, Nt-Tyr, Nt-Leu, Nt-Leu
  • it is an essential determinant of bonding (Sriram and Kwon, 2010).
  • the UBR is an abbreviation of Ubiquitin protein ligase E3 component n-recognin, and UBR is an N-recognin that recognizes an N-terminal degradation signal of a protein. It is known that at least seven types of UBRs 1 to 7 exist in mammals.
  • the UBR box domain common to UBRs is a zinc finger motif having a size of about 70 residues, and is known as a highly conserved substrate-binding domain. [Kwon et al., 1998; Xie and Varshavsky, 1999; Kwak et al., 2004; Varshavsky, 1996; Varshavsky, 1997; Kwon et al., 2011; and Zenker et al., 2014].
  • UBR is an N-recognin associated with the N-terminal pathway law, which is a proteolytic pathway
  • the UBR box domain in UBR is a substrate binding domain.
  • UBR1, UBR2, UBR3 and UBR5 act as ubiquitin protein ligase E3 and are known to have a RING domain or a HECT domain.
  • the N-terminal law substrate binding to the UBR is degraded by the ubiquitin proteasome pathway.
  • the UBR box domain in the UBR recognizes the N-terminal amino acid of the substrate and degrades the substrate through the proteasome pathway by ubiquitination of the substrate through the RING domain or the HECT domain.
  • proteins that are misfolded in cells are degraded through the ubiquitin proteasome pathway because if they are left for a long time, they aggregate and block the proteasome or reduce other cellular functions. (Ji and Kwon, 2017).
  • the UBR box domain plays an important role in the intracellular proteolytic pathway through the recognition of the N-terminal degradation signal.
  • ligands that bind to the UBR box domain can affect intracellular proteolytic pathways.
  • the present specification relates to compounds as ligands that bind to the UBR box domain associated with intracellular proteolytic pathways, as described above.
  • the present specification provides a small molecule compound that binds to the UBR box domain.
  • the UBR box domain includes UBR box domains in UBRs 1 to 7.
  • the small molecule compound may function as a ligand suitable for binding to the UBR box domain.
  • the present specification provides a composition for inhibition of UBR box domain substrate binding comprising a ligand compound binding to the UBR box domain.
  • the present specification provides a pharmaceutical composition for treating UBR-related diseases, including a ligand compound that binds to a UBR box domain, and uses thereof.
  • the present specification includes a ligand compound that binds to the UBR box domain, muscle loss caused by muscular atrophy (Becker, Congennital, Duchenne, Distal, Emery-Dreifuss, Facioscapulohumeral, Limb-girdle, myotonic) , liposarcoma caused by excessive protein breakdown, as well as diseases mediated by muscle loss or degradation, including ocuophargyngeal (muscular dystrophy), sarcopenia or cancer cachexia; UBR box and UBR protein, such as cystic fibrosis, Johanson-Blizzard syndrome, urethral obstruction sequence, autoimmune pancreatitis or Usher syndrome Provided are pharmaceutical compositions and uses thereof for treating diseases known as related diseases.
  • muscular atrophy Becker, Congennital, Duchenne, Distal, Emery-Dreifuss, Facioscapulohumeral, Limb-girdle, myotonic
  • liposarcoma caused by excessive protein breakdown, as
  • the present specification provides a compound having the structure of Formula 1 or a salt thereof.
  • a 1 is CH 3 or NH 2 .
  • a 1 is CH 3 or NH 2 It provides a composition for inhibiting UBR box domain substrate binding comprising a compound or a salt thereof.
  • A1 is CH 3 or NH 2
  • a pharmaceutical composition for treating a UBR-related disease comprising a compound or a pharmaceutically acceptable salt thereof, and a method for treating a UBR-related disease using the same.
  • the UBR-related disease is muscle loss (Becker, Congennital, Duchenne, Distal, Emery-Dreifuss, Facioscapulohumeral, Limb-girdle, myotonic, ocuophargyngeal) (muscular dystrophy), sarcopenia or cancer cachexia caused by muscular atrophy.
  • muscle wasting diseases including cancer cachexia, liposarcoma, cystic fibrosis, and Johanson-Blizzard syndrome caused by excessive protein breakdown ), urethral obstruction sequence, autoimmune pancreatitis, or Usher syndrome.
  • One invention disclosed herein is to provide a ligand compound having a higher binding affinity to the UBR box domain.
  • UBR box domain substrate Binding of a UBR box domain substrate can be inhibited through a UBR box domain ligand compound, and various uses utilizing this property can be provided.
  • UBR-related diseases eg, muscle loss, etc.
  • UBR box domain ligand compound can be treated via a UBR box domain ligand compound.
  • UBR Ubiquitin protein ligase E3 component n-recognin
  • the term UBR refers to an abbreviation of Ubiquitin protein ligase E3 component n-recognin.
  • the UBR is an N-recognin that recognizes an N-terminal residue of a protein, and it is known that at least seven types of UBR 1 to 7 exist in mammals.
  • the UBR is N-recognin, and is related to the N-terminal law pathway, which is an in vivo proteolytic pathway. Specifically, the UBR recognizes the N-terminal degradation signal (N-degron) of the protein, and is involved in the degradation of the substrate protein through the ubiquitin proteasome pathway.
  • N-degron N-terminal degradation signal
  • UBR box domain is a domain present in a UBR protein and is a zinc finger motif.
  • the UBR protein includes UBR 1 to 7 proteins.
  • the UBR box domain is known as a domain to which a substrate protein binds.
  • the compounds disclosed herein as UBR box domain ligands can bind to the UBR box domain and inhibit the binding of a UBR box domain substrate.
  • compounds as UBR box domain ligands disclosed herein may affect intracellular proteolytic pathways.
  • RING domain is known to be present in UBR 1, 2 and 3 proteins.
  • the RING domain may also be used as the term RING ubiquitination domain.
  • the RING domain is a domain present in a protein and is a zinc finger motif.
  • the RING domain is a domain that plays an important role in the process in which ubiquitin in E2 is moved to a substrate protein, and the RING domain serves to cause the process in which the ubiquitin is transferred to a substrate protein occurs in one-step.
  • HECT domain is known to be present in the UBR 5 protein.
  • the HECT domain may also be used as the term HECT ubiquitination domain.
  • the HECT domain is a domain that plays an important role in the process by which ubiquitin in E2 is transferred to a substrate protein. Ubiquitin in E2 is delivered to the HECT domain and then translocated to the matrix protein. That is, the HECT domain serves to cause the transfer of the ubiquitin to the substrate protein in two steps.
  • zinc finger motif refers to a protein structure motif in which one or more zinc (Zinc) ions are present in order to stabilize the structure of the protein.
  • Zinc zinc ions
  • the term ligand refers to a substance that specifically binds to a protein.
  • the protein includes an enzyme or a receptor, and when the protein is an enzyme, the ligand may refer to a substrate binding to an enzyme, and when the protein is a receptor, the ligand may refer to a hormone binding to a receptor.
  • a compound as a UBR box domain ligand provided herein refers to a compound that binds to a UBR box domain.
  • the compound refers to a compound that binds to the UBR box domain in the UBR protein.
  • the compound refers to a compound that binds to the UBR box domain present in one or more proteins of UBR1 to 7.
  • the present invention is not limited thereto.
  • the compounds provided herein as UBR box domain ligands can act competitively with substrates of the UBR box domain. That is, the compound can inhibit the binding of the substrate of the UBR box domain. In addition, the compound may inhibit the decomposition of the substrate by inhibiting the binding of the substrate.
  • UBR box domain ligands bind to the UBR box domain.
  • the UBR box domain is known as a domain to which an N-terminal residue sequence or an N-terminal degradation signal binds. This domain is involved in the degradation of proteins by the law of the N-terminal pathway. Thus, the compound may affect the proteolytic process via the N-terminal law pathway.
  • N-degron is a degradation signal of the protein.
  • protein degradation is regulated depending on the N-terminal residue sequence of the protein, and the protein degradation signals present at the N-terminus are collectively referred to as N-degron.
  • the N-degron includes those having a positively charged residue (eg, arginine, lysine, histidine) or a large hydrophobic residue (phenylalanine, leucine, tryptophan, isoleucine, tyrosine) at the N-terminus.
  • the term N-end rule was used based on the association that the half-life of a protein is determined by what amino acid residues exist at the N-terminus of the protein.
  • N-degron is recognized by N-recognin
  • UBR Ubiquitin protein ligase E3 component n-recognin
  • the protein degradation process by the UBR may include the following.
  • the UBR box domain recognizes a substrate having an N-terminal degradation signal, ubiquitin is bound to the substrate, and the substrate to which ubiquitin is bound can be degraded by the proteasome. That is, a substrate having an N-terminal degradation signal can be degraded by the ubiquitin proteasome system (UPS).
  • UPS ubiquitin proteasome system
  • the compounds of the present specification reflect the structure of the UBR box domain and the binding properties of the N-terminal pathway substrate.
  • the compounds as UBR box domain ligands disclosed herein were designed in consideration of the structure of the UBR box domain and the binding form between the UBR box domain and the N-terminal pathway substrate.
  • Various amino acids present in the UBR box domain interact and bind with amino acids of the N-terminal pathway substrate through ionic interactions, hydrogen bonds, hydrophobic interactions, and the like.
  • a small molecule compound capable of forming a suitable binding mode with the UBR box domain is synthesized and provided.
  • a compound represented by Formula 1 is provided below.
  • the compound of the present specification has a structure that binds well to the UBR box domain
  • a 1 is CH 3 or NH 2 .
  • the compound of Formula 1 disclosed herein may exist in the form of a stereoisomer or salt thereof, and the isomer or salt form of such a compound is included in the scope of the present specification.
  • the UBR box domain ligand compounds disclosed herein may be selected from:
  • the compound may be considered in the form of one of its possible isomers or a mixture thereof.
  • all stereoisomers or mixtures thereof eg racemic mixtures, including enantiomers and diastereomers, are contemplated.
  • salts include pharmaceutically acceptable salts.
  • Salts disclosed herein include acid addition salts or basic addition salts.
  • Exemplary acids that form the salts include hydrochloric acid, sulfuric acid, phosphoric acid, glycolic acid, lactic acid, pyruvic acid, citric acid, succinic acid, glutaric acid, and the like
  • exemplary bases that form salts include lithium, sodium, potassium, calcium, magnesium. , methylamine, trimethylamine, and the like.
  • the present invention is not limited thereto and may be easily selected by those skilled in the art.
  • compositions for inhibiting binding of UBR box domain substrates can be used to prepare compositions for inhibiting binding of UBR box domain substrates.
  • a composition comprising the compound disclosed herein may be used to inhibit binding of a UBR box domain substrate by binding to a UBR box domain.
  • the composition including the compound may be used for preventing degradation of a substrate that is bound to the UBR box domain and degraded.
  • the composition including the compound may be used for preventing the substrate from being bound to the UBR box domain and degraded by the ubiquitin-proteasome pathway.
  • the composition comprising the compound disclosed herein may be used for inhibiting the binding of a substrate having an N-terminal residue binding to the UBR box domain.
  • N- such as arginine (Arg), lysine (Lys), histidine (His), tryptophan (Trp), phenylalanine (Phe), tyrosine (Tyr), leucine (Leu), and isoleucine (Ile) It can be used for the purpose of inhibiting the binding of a substrate having a terminal residue.
  • the present invention is not limited thereto, and may be used for inhibiting the binding of a substance known as a substrate of the UBR box domain in the art.
  • the compound of the present specification or a salt thereof has a property of binding to the UBR box domain. That is, it is a compound that functions as a ligand that binds to the UBR box domain. Therefore, using such a compound, it is possible to inhibit the degradation of proteins that are degraded through binding to the UBR box domain in the body, and UBR-related diseases can be treated using this mechanism.
  • the compounds disclosed herein can be used to prepare pharmaceutical compositions for treating a subject in need thereof.
  • the treatment includes improving the symptoms of a specific medical condition or having the effect of delaying the progression of a disease.
  • the subject includes humans and non-human animals.
  • the pharmaceutical composition may include a pharmaceutically acceptable carrier, excipient and/or additive together with the compound.
  • the pharmaceutically acceptable carriers, excipients and/or additives include, but are not limited to, water, saline, glycol, glycerol, animal and vegetable fats, oils, starches, and the like, pharmaceutically acceptable carriers, excipients known in the art. and/or additives.
  • a method of treatment comprising administering to a subject in need thereof a compound disclosed herein, or a pharmaceutically acceptable salt thereof.
  • the administration of the compound or a pharmaceutically acceptable salt thereof may have the effect of improving symptoms of a specific medical condition or delaying the progression of a disease compared to a subject who has not received the administration.
  • the subject includes humans and non-human animals.
  • the present specification provides a treatment method comprising administering the compound or a pharmaceutically acceptable salt thereof to a subject having a UBR-related disease. That is, the compound disclosed herein or a pharmaceutically acceptable salt thereof may be used to treat a UBR-related disease. In a specific embodiment, the compound or a pharmaceutically acceptable salt thereof may be used to treat a specific disease that can be treated by inhibiting the degradation of a protein that is degraded through binding to the UBR box domain.
  • the specific disease is muscle loss caused by muscle atrophy (Becker, Congennital, Duchenne, Distal, Emery-Dreifuss, Facioscapulohumeral, Limb-girdle, myotonic, ocuophargyngeal) (muscular dystrophy) or sarcopenia or cancer cachexia cachexia), along with muscle wasting diseases, including liposarcoma, cystic fibrosis, and Johanson-Blizzard syndrome, caused by excessive protein breakdown , urethral obstruction sequence, autoimmune pancreatitis, and Usher syndrome.
  • the compound or a pharmaceutically acceptable salt thereof may be used to treat muscle loss mediated by UBR.
  • the rapid loss of muscle mass accompanying disease states such as cancer, sepsis, and hyperthyroidism is associated with increased protein breakdown in muscle, which is known to be associated with activation of the ubiquitin proteasome system.
  • ubiquitin binding is increased and consequently muscle loss occurs through activation of the N-terminal law pathway in particular [ALFRED L. GOLDBERG et al. 1998, 1999].
  • a compound disclosed herein, or a pharmaceutically acceptable salt thereof can be used to treat the disease by preventing activation of the muscle loss pathway through binding to the UBR box domain.
  • the specific disease includes all diseases known as diseases related to UBR in the art.
  • a rat muscle-derived cell After culturing the L6 cell line, a rat muscle-derived cell, using DMEM medium containing 10% FBS and 1% streptomycin/penicillin in an incubator maintained with 5% carbon dioxide, each cell was seeded in a 12-well plate. Additional incubation was carried out for 24 hours so that the cells were completely attached to the surface of the plate. To determine whether MG132 increases UBR1 binding, cells were collected after treatment with MG132 (10 uM) alone for 24 hours.
  • lysis buffer (20 mM Tris, pH 7.4, 150 mM NaCl, 1% triton-X-100, 2 mM NaF, 2 mM EDTA, 2 mM b-glycerophosphate, 5 mM sodium orthovanadate, 1 mM PMSF, leupeptin, aprotenin
  • a sample buffer was added to each sample and reacted at 100 °C for 5 minutes.
  • 5 uL of the reaction sample was taken and dispensed into each well of an acrylamide gel, followed by immunoblotting, and the experimental results are shown in FIG. 1 .
  • the immunoblotting method is schematically representative from three or more independent experiments.
  • the levels of ACTA1, ACTC1, and ACTG2 are further increased by MG132 than in the control group.
  • the level of ACTA1 and ACTG2 increased when the UBR protein was knocked down. That is, it was confirmed that muscle actin is an Arg/N-degron pathway substrate.
  • the TnT® Quick Coupled Transcription/Translation System kit was used.
  • Transcend Biotin-Lysyl-tRNA , Methionine, Bestatin, TnT quick Master mix and DHFR-Ub-R-nsP4 Plasmid were used to make a Pre-mix and then mixed with the compound (1 uM). After incubating each sample at 30 °C for 40 minutes, 5X SDS loading dye was added. After reacting at 95 °C for 2 minutes, 5 uL was taken and dispensed into each well of acrylamide gel, followed by immunoblotting, and the results of the experiment are shown in [Figure 2].
  • the in vitro transcriptional translation method is schematically representative from at least three independent experiments.
  • the level of R-nsP4 is further increased by compounds 1 and 2 compared to the control group. That is, when the compound according to the present invention was treated, it was confirmed that the R-nsP4 level was increased through binding to UBR1.
  • the L6 cell line a rat muscle-derived cell
  • DMEM medium containing 10% FBS and 1% streptomycin/penicillin
  • each cell was seeded in a 12-well plate. An additional 24 hours of incubation was performed so that the cells were completely attached to the surface of the plate.
  • the cells were collected after treatment with the compound (5 uM) alone for 24 hours.
  • lysis buffer (20 mM Tris, pH 7.4, 150 mM NaCl, 1% triton-X-100, 2 mM NaF, 2 mM EDTA, 2 mM b-glycerophosphate, 5 mM sodium orthovanadate, 1 mM PMSF, leupeptin, aprotenin
  • a sample buffer was added to each sample and reacted at 100 °C for 5 minutes.
  • 5 uL of the reaction sample was taken and dispensed into each well of acrylamide gel, followed by immunoblotting, and the results of the experiment are shown in [Fig. 3].
  • the immunoblotting method was schematically representative from three or more independent experiments.
  • the level of ACTA1 was further increased by compounds 1 and 2 compared to the control group. That is, it can be confirmed that when the compound according to the present invention is treated, it inhibits the degradation of ACTA1, an intramuscular protein, by binding to UBR1.
  • Gln97-Pro168 which corresponds to the UBR box of Human UBR1 (UniProt ID: Q8IWV7), was cloned into a modified expression vector and expressed in E. coli. After affinity chromatography was used to remove the tag with protease, Gly-His-Met was added to the N-terminus. After performing ion chromatography, the final UBR box protein of UBR1 was purified by gel filtration chromatography in a buffer composition of 10 mM NaCl, 20 mM Tris-HCl, 2 mM beta-mercaptoethanol, pH 7.5.
  • the dye of The Monolith Protein Labeling Kit RED-NHS 2nd generation (Cat# MO-L011) has an NHS-ester group that forms a covalent bond with primary amines (lysine residues). This dye is optimized for monolith-series instruments with RED detectors. UBR1 UBR box protein purified using this kit was labeled according to the presented protocol.
  • Thermophoresis refers to the movement of particles by a temperature gradient. Particles in the high-temperature region have greater kinetic energy than particles in the low-temperature region and collide more frequently with surrounding particles with greater energy. As a result, the particles move from the high-temperature region to the low-temperature region.
  • Thermophoresis of proteins is typically different from thermophoresis of protein-ligand complexes. This is because the size, charge, and solvation energy change as the ligand binds. Even though ligand binding does not significantly change the size and charge of the protein, MST can detect changes in the solvation entropy of protein molecules due to ligand binding. Therefore, the binding between the UBR1 UBR box protein and the ligand compound was measured using MST, and it was confirmed that the presented ligand binds to the UBR1 UBR box.

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Abstract

본 명세서는 UBR 박스 도메인 리간드로의 화합물에 관한 것이다. 본 명세서에서는 UBR 박스 도메인에 결합하는 저분자 화합물을 제공한다. 또한, 본 명세서는 UBR 박스 도메인에 결합하는 리간드 화합물을 포함하는 UBR 박스 도메인 기질 결합 억제용 조성물, UBR 관련 질환 치료용 약학적 조성물 및 이의 용도를 제공한다.

Description

UBR 박스 도메인 리간드로의 화합물
[관련 출원에 대한 상호참조]
본 출원은 2020년 4월 27일에 출원된 미국 가특허출원 일련번호 63/015,945의 혜택과 우선권을 주장하며, 그 전체가 여기에 참조로 포함된다.
[기술분야]
본 명세서에서 개시하는 내용은 UBR 박스 도메인 리간드로의 화합물에 관한 것이다. 상기 UBR 박스 도메인은 N-말단 경로 법칙(N-end rule pathway)의 UBR(Ubiquitin protein ligase E3 component n-recognin)단백질 내에 공통적으로 존재하는 도메인이다. 이 때 상기 UBR 박스 도메인은 기질(substrate)이 결합하는 도메인으로 알려져 있다. 상기 UBR 박스 도메인은 기질의 N-말단 잔기에 결합하여 기질에 멀티 유비퀴틴 사슬을 형성하는데 필수적이며, 기질은 이 과정을 통해 분해된다고 알려져 있다.
본 명세서는 상기 UBR 박스 도메인에 결합하는 리간드로 작용하는 화합물에 관한 것이다.
세포는 단백질 분해를 통해 생체 내 단백질의 양과 기능을 조절한다. 이 때, 생체내의 단백질은 N-말단 잔기 서열에 의존적으로 분해될 수 있으며, 이러한 분해 경로는 N-말단 경로 법칙(N-end rule pathway)이라고 알려져 있다. 즉, N-말단 경로 법칙은 특정 단백질 N-말단을 분해신호로 사용하는 단백질 분해 시스템이다. 상기 N-말단 경로 법칙은 아래와 같은 단백질 분해 과정을 포함할 수 있다.
진핵 생물의 경우, N-레코그닌(recognin)이 단백질의 N-말단 분해 신호를 인식하고, 상기 N-레코그닌은 분해 대상 단백질에 유비퀴틴을 결함시킴으로써 단백질을 분해시킬 수 있다. 이때 상기 N-말단 분해 신호는 N-말단에 양전하를 갖는 잔기(타입 1; 예, 아르기닌, 라이신, 히스티딘) 또는 크기가 큰 소수성 잔기(타입 2; 페닐알라닌, 류신, 트립토판, 이소류신, 티로신)를 가지는 것을 포함할 수 있다. 본 발명자들은 N-레코그닌인 UBR 1, UBR2, UBR3, 및 UBR5를 최초로 발견하거나 클로닝 하였으며, 이들이 기질인식 도메인으로 UBR 박스 도메인을 가지고 있음을 밝힌 바 있다(Tasaki et al. 2005). 이 때, N-레코그닌이 N-end rule 리간드와 결합함으로서 만들어진 유비퀴틴화된 기질은 프로테아좀에 전달되어 짧은 펩타이드로 분해된다. 이러한 과정에서 특정 N-말단 잔기(Nt-Arg, Nt-His, Nt-Lys, Nt-Trp, Nt-Phe, Nt-Tyr, Nt-Leu, Nt-Leu)는 N-레코그닌이 N-end rule 기질을 타겟팅할 때 필요한 수소결합(hydrogen bond)를 대부분 제공하기 때문에 결합에 꼭 필요한 결정인자이다(Sriram and Kwon, 2010).
상기 UBR은 Ubiquitin protein ligase E3 component n-recognin의 약자로, UBR은 단백질의 N-말단 분해 신호를 인식하는 N-레코그닌(recognin)이다. 상기 UBR은 포유류 내에 UBR 1 내지 7의 적어도 7가지 종류가 존재한다고 알려져 있다. 또한, UBR이 공통적으로 가지고 있는 UBR 박스 도메인은 약 70 잔기 정도의 크기를 가지는 징크 핑거 모티프(zinc finger motif)이며, 고도로 보존된 기질-결합 도메인으로 알려져 있다. [Kwon et al., 1998; Xie and Varshavsky, 1999; Kwak et al., 2004; Varshavsky, 1996; Varshavsky,1997; Kwon et al., 2011; and Zenker et al., 2014].
즉, UBR은 단백질 분해 경로인 N-말단 경로 법칙과 연관되어 있는 N-레코그닌이며, UBR 내의 UBR 박스 도메인은 기질 결합 도메인이다. 특별히, 상기 UBR 1 내지 7 중에서 UBR1, UBR2, UBR3 및 UBR5는 유비퀴틴 단백질 리가아제 E3 로 작용하며, RING 도메인 또는 HECT 도메인을 가지고 있다고 알려져 있다. 상기 UBR에 결합하는 N-말단 법칙의 기질은 유비퀴틴 프로테아좀 경로에 의해 분해된다. 구체적으로, 상기 UBR 내의 UBR 박스 도메인은 기질의 N-말단 아미노산을 인식하고 RING 도메인이나 HECT 도메인을 통해 기질을 유비퀴틴화 시킴으로써 기질을 프로테아좀 경로를 통해 분해시킨다. 예를 들어, 세포에서 폴딩이 제대로 되지 않은(misfolded) 단백질들이 장시간 방치될 경우 응고되어(aggregated) 프로테아좀을 막거나 다른 세포기능을 저하할 수 있기 때문에, 유비퀴틴 프로테아좀 경로를 통해 분해된다(Ji and Kwon, 2017).
즉, UBR 박스 도메인은 N-말단 분해 신호 인식을 통해, 세포 내 단백질 분해 경로에 중요한 역할을 한다. 따라서, UBR 박스 도메인에 결합하는 리간드는 세포 내 단백질 분해 경로에 영향을 줄 수 있다.
본 명세서는 위에 기재한 바와 같이, 세포 내 단백질 분해 경로와 연관되어 있는 UBR 박스 도메인에 결합하는 리간드로의 화합물에 관한 것이다.
본 명세서는 UBR 박스 도메인에 결합하는 저분자 화합물을 제공한다. 이때, 상기 UBR 박스 도메인은 UBR 1 내지 7 내의 UBR 박스 도메인을 포함한다. 상기 저분자 화합물은 UBR 박스 도메인 결합에 적합한 리간드로 기능할 수 있다.
일 구현예로, 본 명세서에서는 UBR 박스 도메인에 결합하는 리간드 화합물을 포함하는 UBR 박스 도메인 기질 결합 억제용 조성물(a composition for inhibition of UBR box domain substrate binding)을 제공한다.
구체적인 일 구현예로, 본 명세서는 UBR 박스 도메인에 결합하는 리간드 화합물을 포함하는, UBR 관련 질환 치료용 약학적 조성물 및 이의 용도를 제공한다.
보다 구체적인 일 구현예로, 본 명세서는 UBR 박스 도메인에 결합하는 리간드화합물을 포함하는, 근육 위축증으로 발병되는 근육 손실증(Becker, Congennital, Duchenne, Distal, Emery-Dreifuss, Facioscapulohumeral, Limb-girdle, myotonic, ocuophargyngeal)(muscular dystrophy), 근육감소증(sarcopenia)이나 암 악액질(cancer cachexia)을 포함한 근육 손실이나 분해로 매개되는 질병(muscle wasting disease)들과 더불어 과다한 단백질 분해로 발병되는 지방육종(liposarcoma), 낭포성섬유증(cystic fibrosis), 요한슨-블리자드 질병(Johanson-Blizzard syndrome), 폐쇄성 요로질환(urethral obstruction sequence), 자가면역 췌장염(autoimmune pancreatitis) 또는 어셔 질병(Usher syndrome)과 같은 UBR box와 UBR 단백질에 관련된 질환으로 공지된 질환들을 포함하는 치료용 약학적 조성물 및 이의 용도를 제공한다.
본 명세서에서는 화학식 1의 구조를 가지는 화합물 또는 이의 염을 제공한다.
[화학식1]
Figure PCTKR2021005335-appb-I000001
이때, A1은 CH3 또는 NH2이다.
이때, 일 실시예로 상기 A1은 CH3 또는 NH2인 화합물 또는 이의 염을 포함하는 UBR 박스 도메인 기질 결합 억제용 조성물을 제공한다.
이때, 일 실시예로 상기 A1은 CH3 또는 NH2인 화합물 또는 이의 제약상 허용되는 염을 포함하는 UBR 관련 질환 치료용 약학적 조성물 및 이를 이용하는 UBR 관련 질환 치료 방법을 제공한다.
이 때, 일 실시예로, 상기 UBR 관련 질환은 근육 위축증으로 발병되는 근육 손실증(Becker, Congennital, Duchenne, Distal, Emery-Dreifuss, Facioscapulohumeral, Limb-girdle, myotonic, ocuophargyngeal)(muscular dystrophy), 근육감소증(sarcopenia)이나 암 악액질(cancer cachexia)을 포함한 근육 손실이나 분해로 매개되는 질병(muscle wasting disease)들과 더불어 과다한 단백질 분해로 발병되는 지방육종(liposarcoma), 낭포성섬유증(cystic fibrosis), 요한슨-블리자드 질병(Johanson-Blizzard syndrome), 폐쇄성 요로질환(urethral obstruction sequence), 자가면역 췌장염(autoimmune pancreatitis) 또는 어셔 질병(Usher syndrome)에서 선택될 수 있다.
본 명세서에서 개시하고 있는 일 발명은, UBR 박스 도메인에 대해 보다 결합력이 높은 리간드 화합물을 제공하는 것이다.
UBR 박스 도메인 리간드 화합물을 통해 UBR 박스 도메인 기질의 결합이 억제될 수 있고, 이러한 특성을 이용하는 다양한 용도를 제공할 수 있다. 예를 들어, UBR 박스 도메인 리간드 화합물을 통해 UBR 관련 질환(예, 근육 손실증 등)이 치료될 수 있다.
도1은 면역블로팅법을 이용하여 머슬 액틴(muscle actin)이 Arg/N-데그론(degron) 경로 기질인지 여부를 확인한 실험 결과이다.
도2는 시험관 내 전사 번역 방법을 이용하여 화합물(화합물 1, 2)이 UBR1에 결합함에 따른 R-nsP4 발현 여부를 확인한 실험 결과이다.
도3은 면역블로팅법을 이용하여 화합물(화합물 1,2)이 근육 세포 내 액틴 분해를 억제하는지 확인한 실험 결과이다.
도 4 내지 5는 MST에 의한 화합물1과 화합물2의 결합측정 결과이다.
이하, 첨부된 도면을 참조하여, 발명의 내용을 특정한 구현예와 예시들을 통해 더욱 상세하게 설명한다. 상기 첨부된 도면은 발명의 일부 구현예를 포함하지만, 모든 구현예를 포함하고 있지는 않다는 점에 유의해야 한다. 본 명세서에 의해 개시되는 발명의 내용은 다양하게 구현될 수 있으며, 여기에 설명되는 특정 구현예로 제한되지 않는다. 본 명세서에 개시된 발명이 속한 기술분야에 있어 통상의 기술자라면, 본 명세서에 개시된 발명의 내용에 대한 많은 변형 및 다른 구현예들을 떠올릴 수 있을 것이다. 따라서, 본 명세서에서 개시된 발명의 내용은 여기에 기재된 특정 구현예로 제한되지 않으며, 이에 대한 변형 및 다른 구현예들도 청구범위 내에 포함되는 것으로 이해되어야 한다.
용어 정의
본 명세서에서 사용하는 주요 용어에 대한 정의는 아래와 같다.
UBR (Ubiquitin protein ligase E3 component n-recognin)
본 명세서에서 사용되는 용어 UBR은 Ubiquitin protein ligase E3 component n-recognin의 약자를 의미한다. 상기 UBR은 단백질의 N-말단 잔기를 인식하는 N-레코그닌이며, 포유류 내에 UBR 1 내지 7의 적어도 7가지 종류가 존재한다고 알려져 있다. 상기 UBR은 N-레코그닌으로, 생체 내 단백질 분해 경로인 N-말단 법칙 경로에 연관되어 있다. 구체적으로, 상기 UBR은 단백질의 N-말단 분해 신호(N-degron)를 인식하며, 기질 단백질이 유비퀴틴 프로테아좀 경로를 통해 분해되는 과정에 연관되어 있다.
UBR 박스 도메인(UBR box domain)
본 명세서에서 사용되는 용어 UBR 박스 도메인은 UBR 단백질 내에 존재하는 도메인으로 징크 핑거 모티프이다. 상기 UBR 단백질은 UBR 1 내지 7 단백질을 포함한다. 상기 UBR 박스 도메인은 기질(substrate) 단백질이 결합하는 도메인으로 알려져 있다. 본 명세서에 개시되는 UBR 박스 도메인 리간드로의 화합물은 상기 UBR 박스 도메인에 결합하여 UBR 박스 도메인 기질의 결합을 억제할 수 있다. 더 나아가, 본 명세서에서 개시되는 UBR 박스 도메인 리간드로의 화합물은 세포 내 단백질 분해 경로에 영향을 줄 수 있다.
RING 도메인(RING domain)
본 명세서에서 사용되는 용어 RING 도메인은 UBR 1, 2 및 3 단백질 내에 존재한다고 알려져 있다. 상기 RING 도메인은 RING 유비퀴틴화 도메인이라는 용어로도 사용될 수 있다. 상기 RING 도메인은 단백질 내에 존재하는 도메인으로 징크 핑거 모티프이다. 상기 RING 도메인은 E2에 있는 유비퀴틴이 기질 단백질로 이동되는 과정에 중요한 역할을 하는 도메인으로, RING 도메인은 상기 유비퀴틴이 기질 단백질로 이동되는 과정이 한 단계(one-step)로 일어나게 하는 역할을 한다.
HECT 도메인(HECT domain)
본 명세서에서 사용되는 용어 HECT 도메인은 UBR 5 단백질 내에 존재한다고 알려져 있다. 상기 HECT 도메인은 HECT 유비퀴틴화 도메인이라는 용어로도 사용될 수 있다. 상기 HECT 도메인은 E2에 있는 유비퀴틴이 기질 단백질로 이동되는 과정에 중요한 역할을 하는 도메인이다. E2에 있는 유비퀴틴은 HECT 도메인에 전달되고, 그 후 기질 단백질로 이동되게 된다. 즉, HECT 도메인은 상기 유비퀴틴이 기질 단백질로 이동되는 과정이 두 단계(two step)로 일어나게 하는 역할을 한다.
징크 핑거 모티프(zinc finger motif)
본 명세서에서 사용되는 용어 징크 핑거 모티프는 단백질의 구조를 안정시키기 위해 하나 이상의 아연(Zinc) 이온이 존재하는 단백질 구조 모티프를 의미한다. 본 명세서의 UBR 박스 도메인 및 RING 도메인은 징크 핑거 모티프이다.
리간드(ligand)
본 명세서에서 사용되는 용어 리간드는 단백질에 특이적으로 결합하는 물질을 의미한다. 상기 단백질은 효소 또는 수용체를 포함하며, 상기 단백질이 효소인 경우 리간드는 효소에 결합하는 기질 등을 의미할 수 있고, 상기 단백질이 수용체인 경우 리간드는 수용체에 결합하는 호르몬 등을 의미할 수 있다.
본 명세서에서 제공하는 UBR 박스 도메인 리간드로의 화합물은 UBR 박스 도메인에 결합하는 화합물을 의미한다. 일 실시예로, 상기 화합물은 UBR 단백질 내의 UBR 박스 도메인에 결합하는 화합물을 의미한다. 구체적인 일 실시예로, 상기 화합물은 UBR1 내지 7 중 하나 이상의 단백질 내에 존재하는 UBR 박스 도메인에 결합하는 화합물을 의미한다. 다만, 이에 제한되지 않는다.
본 명세서에서 제공하는 UBR 박스 도메인 리간드로의 화합물은 UBR 박스 도메인의 기질과 경쟁적으로 작용할 수 있다. 즉, 상기 화합물은 UBR 박스 도메인의 기질이 결합하는 것을 억제할 수 있다. 또한, 상기 화합물은 상기 기질의 결합을 억제하여 기질의 분해를 저해할 수 있다.
달리 정의되지 않는 한, 본 명세서에서 사용되는 모든 기술적 및 과학적 용어는 본 발명이 속하는 기술분야의 당업자에 의해 통상적으로 이해되는 것과 동일한 의미를 가진다. 본 명세서에서 언급된 모든 간행물, 특허 및 기타 다른 참고문헌은 전체가 참고로 포함된다.
이하, 발명의 구체적인 내용을 개시한다.
Ⅰ. UBR 박스 도메인
1.개요
본 명세서에서 제공하는 UBR 박스 도메인 리간드로의 화합물은 UBR 박스 도메인에 결합한다. 상기 UBR 박스 도메인은 N-말단 잔기 서열 또는 N-말단 분해 신호가 결합하는 도메인으로 알려져 있다. 상기 도메인은 N-말단 경로 법칙에 의해 단백질이 분해되는 과정에 연관되어 있다. 따라서, 상기 화합물은 N-말단 법칙 경로를 통한 단백질 분해 과정에 영향을 미칠 수 있다.
2.N-말단 법칙 경로(N-end rule pathway)
세포들은 단백질 분해를 통해 단백질의 양을 조절한다. 이 때, 단백질의 분해 과정은 단백질의 분해 신호인 데그론(degron)을 인식하는 과정을 통해 진행된다고 알려져 있다. 구체적으로, 단백질의 N-말단 잔기서열에 의존적으로 단백질 분해가 조절되고, N-말단에 존재하는 단백질 분해 신호들을 통틀어 N-데그론(N-degron)이라고 한다. 상기 N-데그론은 N-말단에 양전하를 갖는 잔기(예, 아르기닌, 라이신, 히스티딘) 또는 크기가 큰 소수성 잔기(페닐알라닌, 류신, 트립토판, 이소류신, 티로신)를 가지는 것을 포함한다. 이와 같이 단백질의 반감기는 그 단백질의 N-말단에 존재하는 아미노산 잔기가 무엇인가에 의해 결정된다는 연관성을 기초로 N-말단 법칙(N-end rule)이라는 용어가 사용되었다.
3.UBR 박스 도메인
상기 N-말단 법칙 경로에서 N-데그론은 N-레코그닌에 의해 인식되는데, N-레코그닌으로 UBR(Ubiquitin protein ligase E3 component n-recognin)이 발견되었다. 상기 UBR은 UBR 박스 도메인을 통해 N-말단 잔기 서열 또는 N-말단 분해 신호를 인식한다고 알려져 있다. 즉, UBR이 UBR 박스 도메인을 통해 단백질 분해 신호를 인식하고, 이를 통해 단백질의 분해 과정이 진행된다.
상기 UBR에 의한 단백질 분해 과정은 아래의 내용을 포함할 수 있다. UBR 박스 도메인이 N-말단 분해 신호를 가진 기질을 인식하고, 상기 기질에 유비퀴틴이 결합되고, 유비퀴틴이 결합된 기질은 프로테아좀에 의해 분해될 수 있다. 즉, N-말단 분해 신호를 가진 기질이 유비퀴틴 프로테아좀 시스템(Ubiquitin proteasome system, UPS)에 의해 분해될 수 있다.
Ⅱ. UBR 박스 도메인 리간드
1.본 명세서의 화합물은 UBR 박스 도메인의 구조 및 N-말단 경로 기질과의 결합 특성을 반영한다
본 명세서에서 개시하는 UBR 박스 도메인 리간드로의 화합물은 UBR 박스 도메인의 구조 및 UBR 박스 도메인과 N-말단 경로 기질과의 결합 형태를 고려하여 설계되었다.
UBR 박스 도메인에 존재하는 다양한 아미노산들은 N-말단 경로 기질의 아미노산과 이온 상호작용(ionic interaction), 수소 결합(hydrogen bond), 소수성 작용(hydrophobic interaction) 등으로 상호작용하며 결합한다. 이러한 결합 모드를 분석하여 본 명세서에서는 UBR 박스 도메인과 적합한 결합 모드를 형성할 수 있는 저분자 화합물이 합성되어 제공된다. 나아가, 본 명세서에서는 이하 화학식 1에 의한 화합물이 제공된다.
2.본 명세서의 화합물은 UBR 박스 도메인에 잘 결합하는 구조를 가진다
1)화학식 1
Figure PCTKR2021005335-appb-I000002
[화학식 1]
상기 화학식 1에서 A1은 CH3 또는 NH2이다.
이때, 본 명세서에서 개시되는 화학식 1의 화합물은 이의 입체 이성질체 또는 이의 염의 형태로 존재할 수 있으며, 이와 같은 화합물의 이성질체 또는 염의 형태가 본 명세서의 범위에 포함된다.
2)화합물의 구체적인 예시
i)화합물
아래의 내용은 본 명세서에서 개시되는 화합물의 구체적인 실시예들을 나타낸다.
일 실시예로, 본 명세서에서 개시하는 UBR 박스 도메인 리간드 화합물은 아래로부터 선택될 수 있다:
[화합물 1]
Figure PCTKR2021005335-appb-I000003
,
[화합물 2]
Figure PCTKR2021005335-appb-I000004
.
이때, 상기 화합물은 이의 가능한 이성질체 중 하나의 형태 또는 이의 혼합물의 형태가 고려될 수 있다. 예를 들어, 거울상 이성질체(enantiomer) 및 부분 입체 이성질체(diastereomer)를 포함하는 모든 입체 이성질체(stereoisomer) 또는 이의 혼합물(예, 라세미 혼합물)이 고려될 수 있다.
ii)상기 화합물의 염
본 명세서에서 개시되는 화합물은 이의 염의 형태가 고려될 수 있다. 이때, 염은 제약상 허용되는 염을 포함한다. 본 명세서에서 개시되는 염은 산 부가염(acid addition salt) 또는 염기 부가염(basic addition salt)을 포함한다. 상기 염을 형성하는 예시적인 산은 염산, 황산, 인산, 글리콜산, 락트산, 피루브산, 시트르산, 숙신산, 글루타르산 등을 포함하고, 염을 형성하는 예시적인 염기는 리튬, 나트륨, 칼륨, 칼슘, 마그네슘, 메틸아민, 트리메틸 아민 등을 포함한다. 다만, 이에 제한되지 않고 당업자에 의해 쉽게 선택될 수 있다.
Ⅲ. 화합물의 용도
1.UBR 박스 도메인 기질 결합 억제
UBR 박스 도메인 기질 결합 억제용 조성물(A composition for inhibition of UBR box domain substrate binding)
본 명세서에 개시되는 화합물은 UBR 박스 도메인 기질의 결합 억제를 위한 조성물 제조에 사용될 수 있다. 일 실시예로, 본 명세서에 개시되는 상기 화합물을 포함하는 조성물은 UBR 박스 도메인에 결합하여 UBR 박스 도메인 기질의 결합을 억제하는데 사용될 수 있다. 또 다른 실시예로, 상기 화합물을 포함하는 조성물은 UBR 박스 도메인에 결합되어 분해되는 기질이 분해되는 것을 방지하기 위한 용도로 사용될 수 있다. 구체적인 일 실시예로, 상기 화합물을 포함하는 조성물은 기질이 UBR 박스 도메인에 결합되어 유비퀴틴-프로테아좀 경로에 의해 분해되는 것을 방지하기 위한 용도로 사용될 수 있다.
구체적인 일 실시예로, 본 명세서에 개시되는 화합물을 포함하는 조성물은 UBR 박스 도메인에 결합하는 N-말단 잔기를 가지는 기질의 결합을 억제하는 용도로 사용될 수 있다. 구체적인 일 실시예로, 아르기닌(Arg), 라이신(Lys), 히스티딘(His), 트립토판(Trp), 페닐알라닌(Phe), 티로신(Tyr), 류신(Leu), 및 이소류신(Ile) 등의 N-말단 잔기를 가지는 기질의 결합을 억제하는 용도로 사용될 수 있다. 다만 이에 제한되지 않으며, 당업계에 UBR 박스 도메인의 기질로 공지되어 있는 물질의 결합을 억제하는 용도로 사용될 수 있다.
실시예를 참고하면, 본 명세서에서 개시하는 화합물은 UBR에 결합함을 통해 기질의 분해를 억제하는 것을 확인할 수 있다(도 1, 2 및 3 참고).
2.UBR 관련 질환의 치료
본 명세서의 화합물 또는 이의 염은 UBR 박스 도메인에 결합하는 특성을 가진다. 즉, UBR 박스 도메인에 결합하는 리간드로서 기능하는 화합물이다. 따라서, 이러한 화합물을 이용하여, 체내에서 UBR 박스 도메인에 결합됨을 통해 분해되는 단백질의 분해를 억제할 수 있고, 이러한 메커니즘을 이용하여 UBR 관련 질환을 치료할 수 있다.
1)약학적 조성물
본 명세서에 개시되는 화합물은 이를 필요로 하는 대상을 치료하기 위한 약학적 조성물 제조에 이용될 수 있다.
이때, 상기 치료는 특정한 의학적 상태의 증상이 개선되거나, 질병의 진행이 지연되는 효과를 가지는 것을 포함한다. 이때, 상기 대상은 인간, 비인간 동물을 포함한다. 이때, 상기 약학적 조성물은 상기 화합물과 함께 제약상 허용되는 담체, 부형제 및/또는 첨가제를 포함할 수 있다. 상기 제약상 허용되는 담체, 부형제 및 또는 첨가제는 물, 식염수, 글리콜, 글리세롤, 동물성 및 식물성 지방, 오일, 전분 등을 포함하지만, 이에 제한되지 않고 당업계에 공지되어 있는 제약상 허용되는 담체, 부형제 및/또는 첨가제를 모두 포함한다.
2)치료 방법
본 명세서에서는 본 명세서에 개시되는 화합물 또는 이의 제약상 허용되는 염을 이를 필요로 하는 대상에게 투여하는 것을 포함하는 치료방법을 제공한다. 이 때, 상기 화합물 또는 이의 제약상 허용되는 염의 투여는 투여 받지 않은 대상에 비해 특정한 의학적 상태의 증상이 개선되거나, 질병의 진행이 지연되는 효과를 가질 수 있다. 이때, 상기 대상은 인간, 비인간 동물을 포함한다.
-UBR 관련 질환
일 실시예로, 본 명세서에서는 상기 화합물 또는 이의 제약상 허용되는 염을 UBR 관련 질환을 가지는 대상에게 투여하는 것을 포함하는 치료 방법을 제공한다. 즉, 본 명세서에서 개시하는 화합물 또는 이의 제약상 허용되는 염은 UBR 관련 질환을 치료하는데 사용될 수 있다. 구체적인 일 실시예로, 상기 화합물 또는 이의 제약상 허용되는 염은 UBR 박스 도메인에 결합됨을 통해 분해되는 단백질의 분해를 억제함으로써 치료될 수 있는 특정 질환을 치료하는데 사용될 수 있다.
상기 특정 질환은 근육 위축증으로 발병되는 근육 손실증(Becker, Congennital, Duchenne, Distal, Emery-Dreifuss, Facioscapulohumeral, Limb-girdle, myotonic, ocuophargyngeal)(muscular dystrophy)이나 근육감소증(sarcopenia)이나 암 악액질(cancer cachexia)을 포함한 근육 손실이나 분해로 매개되는 질병(muscle wasting disease)들과 더불어 과다한 단백질 분해로 발병되는 지방육종(liposarcoma), 낭포성섬유증(cystic fibrosis), 요한슨-블리자드 질병(Johanson-Blizzard syndrome), 폐쇄성 요로질환(urethral obstruction sequence), 자가면역 췌장염(autoimmune pancreatitis), 어셔 질병(Usher syndrome)을 포함한다. 일 실시예로, 상기 화합물 또는 이의 제약상 허용되는 염은 UBR에 의해 매개되는 근육 손실을 치료하는데 사용될 수 있다. 예를 들어, 암, 패혈증, 갑상선 기능 항진증 등과 같은 질병 상태에서 수반되는 급격한 근육량의 손실은 근육 내 단백질의 분해가 증가하는 것과 관련이 있고, 이는 유비퀴틴 프로테아좀 시스템의 활성화와 연관되어 있다고 알려져 있다. 이 때, 특별히 N-말단 법칙 경로가 활성화됨을 통해 유비퀴틴 결합이 증가되고 결과적으로 근육 손실이 일어난다고 알려져있다[ALFRED L. GOLDBERG et al. 1998, 1999]. 이에 따라, 본 명세서에 개시되는 화합물 또는 이의 제약상 허용되는 염은 UBR 박스 도메인에 결합함을 통해 근육 손실 경로가 활성화되는 것을 방지하여 상기 질환을 치료하는데 사용될 수 있다. 다만, 이에 제한되지 않고, 상기 특정 질환은 당업계에서 UBR에 관련된 질환으로 공지된 질환들을 모두 포함한다.
Ⅳ. 실시예
실시예 1. 화합물 합성
화합물 목록
화합물 번호 화합물 명칭
1
Figure PCTKR2021005335-appb-I000005
N-(5-아세틸-2-클로로페닐)-5-메톡시펜탄아미드
2
Figure PCTKR2021005335-appb-I000006
4-클로로-3-(5-메톡시펜탄아미도)벤즈아미드
1H NMR 스펙트럼은 Bruker Avance III 400 MHz 및 Bruker Fourier 300 MHz에서 기록되었으며 TMS는 내부 표준으로 사용되었다.LCMS는 Agilent 1260HPLC 및 6120MSD에서 사중극자 질량 분석기(quadrupole Mass Spectrometer)에서 측정되었다. (ES (+) 또는 (-) 이온화모드에서 작동하는 컬럼: C18 (50 Х 4.6mm, 5μm); T = 30oC; 유속 = 1.5 mL/min; 감지된 파장: 220nm, 254nm)
실시예 1-1. 화합물1 (N-(5-아세틸-2-클로로페닐)-5-메톡시펜탄아미드)의 제조
Figure PCTKR2021005335-appb-I000007
단계 1) B2의 합성
MeOH (100 mL) 내 B1 (테트라히드로-2H-피란-2-온, 5.00 g, 50 mmol, 1.0 eq) 및 트리메틸 오르토포르메이트(10.6 g, 100 mmol, 2.0 eq)의 혼합물에 conc. H2SO4(3mL)가 25℃에서 적가되었다. 혼합물이 65℃에서 16시간 동안 교반되었다. 용매가 감압하에 농축되었다. 8 M 수산화 나트륨 수용액(10 mL)이 첨가되고, 혼합물이 80℃에서 2시간 동안 교반되었다. 10℃로 냉각된 후, 반응 혼합물이 4N HCl로 산성화되고 EA(에틸 아세테이트)로 추출되었다 (50 mL x 3). 합한 유기층이 염수로 세척되고, Na2SO4로 건조되고 농축되어 B2 (5-메톡시펜탄산, 4.0 g, yield 60%)가 무색 오일로 얻어졌다.
1HNMR (CDCl3, 400 MHz): δ 3.41 (t, J = 6.0 Hz, 2H), 3.34 (s, 3H), 2.39 (t, J = 7.6 Hz, 2H), 1.62-1.74 (m, 4H).
단계 2) 화합물1의 합성
DMF (5 mL) 내 B2 (5-메톡시펜탄산, 0.2 g, 1.52 mmol, 1.0 eq.) 및 HATU (0.69 g, 1.82 mmol, 1.2 eq.)의 혼합물이 25℃에서 30분 동안 교반되었다. 이어서 1-(3-아미노-4-클로로페닐)에탄-1-온 (0.26 g, 1.52 mmol, 1.0 eq.) 및 트리에틸아민(0.31 g, 0.30 mmol, 2.0 eq)이 첨가되었다. 혼합물이 50℃에서 48시간 동안 교반되었다. 혼합물이 물(50 mL)에 부어지고 EA(에틸 아세테이트)로 추출되었다(50 mL x 3). 합한 유기층이 염수로 세척되고, Na2SO4로 건조되고 농축되어 조생성물이 얻어졌고, prep-HPLC로 정제되었다. 수집된 분획이 농축되어 대부분의 CH3CN가 제거되었다. 잔류 분획이 동결 건조되어 화합물1 (N-(5-아세틸-2-클로로페닐)-5-메톡시펜탄아미드, 38 mg, yield 8.8 %)이 백색 고체로 얻어졌다.
1HNMR (CDCl3, 400 MHz): δ 9.03 (s, 1H), 7.84 (s, 1H), 7.67 (dd, J = 8.4 Hz, 2 Hz, 1H), 7.46 (d, J = 8.4 Hz, 1H), 3.45 (t, J = 6.4 Hz, 2H), 3.35 (s, 3H), 2.61 (s, 3H), 2.52 (t, J = 7.2 Hz, 2H), 1.85-1.88 (m, 2H), 1.69-1.73 (m, 2H).
LCMS ; Mass Calcd.:283.7; MS Found: 283.9.
실시예 1-2. 화합물2 (4-클로로-3-(5-메톡시펜탄아미도) 벤즈아미드)의 제조
Figure PCTKR2021005335-appb-I000008
DMF (5 mL) 내 B2 (5-메톡시펜탄산, 0.2 g, 1.52 mmol, 1.0 eq) 및 HATU (0.69 g, 1.82 mmol, 1.2 eq)의 혼합물이 25℃에서 30분 동안 교반되었다. 이어서 3-아미노-4-클로로벤즈아미드(0.26 g, 1.52 mmol, 1.0 eq) 및 트리에틸아민(0.31 g, 0.30 mmol, 2.0 eq)이 첨가되었다. 혼합물이 50℃에서 48시간 동안 교반되었다. 혼합물이 물(50 mL)에 부어지고 EA(에틸 아세테이트)로 추출되었다(50 mL x 3). 합한 유기층이 염수로 세척되고, Na2SO4로 건조되고 농축되어 조생성물이 얻어졌고, prep-HPLC로 정제되었다. 수집된 분획이 농축되어 대부분의 CH3CN이 제거되었다. 잔류 분획이 동결-건조되어 화합물2 (4-클로로-3-(5-메톡시펜탄아미도)벤즈아미드, 48 mg, yield 11.1%)가 노란색 고체로 얻어졌다.
1HNMR (CDCl3, 400 MHz): δ 8.82 (s, 1H), 7.89 (s, 1H), 7.64 (dd, J = 8.4 Hz, 2 Hz, 1H), 7.46 (d, J = 8.4 Hz, 1H), 5.93-6.38 (m, 2H), 3.45 (t, J = 6.0 Hz, 2H), 3.36 (s, 3H), 3.52 (t, J = 7.2 Hz, 2H), 1.81-1.88 (m, 2H), 1.66-1.73 (m, 2H).
LCMS ; Mass Calcd.:284.7; MS Found: 284.9.
실시예 2. Binding Assay 실험
실시예 2-1 면역 블로팅법을 통해 머슬 액틴(muscle actin)이 Arg/N-데그론(degron) 경로 기질인지 여부 확인
랫드 근육 유래 세포인 L6 세포주를 5% 이산화탄소가 유지되는 배양기내에서 10% FBS와 1% 스트렙토마이신/페니실린을 함유한 DMEM 배지를 사용하여 배양 후 12웰 플레이트에 각각의 세포를 분주하였다. 세포가 플레이트의 표면에 완전히 부착되도록 24시간이 추가적인 배양을 하였다. MG132가 UBR1 결합을 증가시키는지 확인하기 위하여 24시간 동안 MG132(10 uM)을 단독으로 처리한 후 세포를 취합하였다. 취합된 세포로부터 단백질을 추출하기 위해 각 샘플에 50 uL의 용해버퍼(20 mM Tris, pH 7.4, 150 mM NaCl, 1% triton-X-100, 2mM NaF, 2 mM EDTA, 2 mM b-glycerophosphate, 5 mM sodium orthovanadate, 1 mM PMSF, leupeptin, aprotenin)을 주입하고 세포를 용해시켰다. 측정된 총 단백질의 농도를 기반으로 각 샘플에 샘플버퍼를 추가하여 100°C에서 5분간 반응시켰다. 반응을 마친 샘플로부터 5 uL을 취하여 아크릴아마이드 젤의 각 웰에 분주한 후 면역블로팅법을 실시하였고, 실험의 결과는 [도 1]에 나타내었다. 면역블로팅법은 3회 이상의 독립적인 실험으로부터 대표적인 것을 도식 화하였다.
도1을 참고하면, 대조군(Control) 보다 MG132에 의해 ACTA1, ACTC1, ACTG2의 레벨이 더 증가함을 확인할 수 있다. 또한, UBR 단백질을 넉다운(knock down) 하였을 때 ACTA1, ACTG2 레벨이 증가함을 확인할 수 있었다. 즉, 머슬 액틴(Muscle actin)이 Arg/N-데그론 경로 기질인 것을 확인할 수 있었다.
실시예 2-2 시험관 내 전사 번역 방법을 통한 R-nsP4 분해 억제 확인
화합물들의 R-nsP4 발현을 확인하기 위해 TnT® Quick Coupled Transcription/Translation System kit를 사용하였다. Transcend Biotin-Lysyl-tRNA , Methionine, Bestatin, TnT quick Master mix와 DHFR-Ub-R-nsP4 Plasmid를 사용해 Pre-mix를 만든 후 화합물(1 uM)과 섞어주었다. 각 샘플을 30°C에서 40분간 반응시킨 후 5X SDS loading dye를 넣어주었다. 95°C에서 2분간 반응시킨 후 5 uL을 취하여 아크릴아마이드 젤의 각 웰에 분주한 후 면역블로팅법을 실시하였고, 실험의 결과는 [도 2]에 나타내었다. 시험관 내 전사 번역 방법은 3회 이상의 독립적인 실험으로부터 대표적인 것을 도식화하였다.
도2를 참고하면, 대조군(Control)에 비해 화합물 1, 2에 의해 R-nsP4의 레벨이 더 증가함을 확인할 수 있다. 즉, 본 발명에 따른 화합물을 처리한 경우 UBR1에 결합함을 통해 R-nsP4 레벨이 증가함을 확인할 수 있었다.
실시예 2-3 면역블로팅법을 통한 근육 세포 액틴 분해 억제 평가
화합물들이 근육 세포내 액틴 분해를 평가하기 위해 랫드 근육 유래 세포인 L6 세포주를 5% 이산화탄소가 유지되는 배양기내에서 10% FBS와 1% 스트렙토마이신/페니실린을 함유한 DMEM 배지를 사용하여 배양하였다. 본 화합물들 중 선택된 대표 화합물의 처리에 따른 UBR1 결합력을 측정하기 위하여, 12웰 플레이트에 각각의 세포를 분주하였다. 세포가 플레이트의 표면에 완전히 부착되도록 24시간이 추가적인 배양을 하였다. 화합물이 UBR1 결합을 증가시키는지 확인하기 위하여 24시간 동안 화합물(5 uM)을 단독으로 처리한 후 세포를 취합하였다. 취합된 세포로부터 단백질을 추출하기 위해 각 샘플에 50 uL의 용해버퍼(20 mM Tris, pH 7.4, 150 mM NaCl, 1% triton-X-100, 2mM NaF, 2 mM EDTA, 2 mM b-glycerophosphate, 5 mM sodium orthovanadate, 1 mM PMSF, leupeptin, aprotenin)을 주입하고 세포를 용해시켰다. 측정된 총 단백질의 농도를 기반으로 각 샘플에 샘플버퍼를 추가하여 100°C에서 5분간 반응시켰다. 반응을 마친 샘플로부터 5 uL을 취하여 아크릴아마이드 젤의 각 웰에 분주한 후 면역블로팅법을 실시하였고, 실험의 결과는 [도 3]에 나타내었다. 면역블로팅법은 3회 이상의 독립적인 실험으로부터 대표적인 것을 도식 화하였다.
도3을 참고하면, 대조군(Control)에 비해 화합물 1, 2에 의해 ACTA1의 레벨이 더 증가함을 확인할 수 있었다. 즉, 본 발명에 따른 화합물을 처리한 경우 UBR1에 결합함을 통해 근육 내 단백질인 ACTA1 분해 억제함을 확인할 수 있다.
실시예 2-4 MST(Microscale thermophoresis)를 이용한 UBR1과 화합물의 결합여부 측정
1) UBR1 단백질 준비
Human UBR1 (UniProt ID : Q8IWV7)의 UBR 박스에 해당하는 Gln97-Pro168부분을 modified expression vector에 클로닝한 후 E.coli에서 발현하였다. 친화 크로마토그래피를 사용 후 protease로 tag을 제거한 후 N-말단에 Gly-His-Met이 추가되었다. 이온 크로마토그래피를 수행한 후, 10 mM NaCl, 20 mM Tris-HCl, 2 mM beta-mercaptoethanol, pH 7.5의 버퍼 조성에서 겔 여과 크로마토그래피를 사용하여 최종 UBR1의 UBR 박스 단백질을 정제하였다.
2) UBR1 UBR 박스 단백질 레이블링
The Monolith protein labeling Kit RED-NHS 2nd generation (Cat# MO-L011)의 dye는 primary amines (lysine residues)과 공유결합을 형성하는 NHS-ester group을 가지고 있다. 이 dye는 RED detector를 가진 Monolith-series 장비에 최적화 되어있다. 이 kit를 이용하여 정제된 UBR1 UBR 박스 단백질은 제시된 프로토콜을 따라 단밸질을 표지했다.
3) MST를 이용하여 UBR1과 ligand 결합여부 측정
열 영동(thermophoresis)은 온도 구배에 의해 입자가 이동하는 현상을 말한다. 높은 온도 영역에 있는 입자는 낮은 온도 영역에 있는 입자보다 더 큰 운동에너지를 가지고 주변 입자들과 더 큰 에너지로 더 자주 충동하게 된다. 그 결과 입자들은 높은 온도 영역에서 낮은 온도 영역으로 이동하게 된다.
단백질의 열 영동은 단백질-ligand 복합체의 열 영동과는 전형적으로 다르다. 리간드가 결합함으로써 크기, 전하 그리고 용매화 에너지(solvation energy)가 변하기 때문이다. 심지어 리간드 결합이 단백질의 크기, 전하를 중요하게 변하지 않게 하더라도, MST는 리간드 결합에 의한 단백질 분자의 용매화 엔트로피(solvation entropy) 변화를 감지할 수 있다. 따라서 UBR1 UBR 박스 단백질과 리간드 화합물과의 결합여부를 MST를 이용하여 측정하였고 제시된 리간드는 UBR1 UBR 박스와 결합함을 확인하였다.

Claims (4)

  1. 화학식 1의 구조를 가지는 화합물:
    [화학식 1]
    Figure PCTKR2021005335-appb-I000009
    또는 이의 염,
    이때 A1은 CH3 또는 NH2인 화합물 또는 이의 염.
  2. 제1항에 있어서,
    상기 A1은 CH3 또는 NH2인 화합물 또는 이의 염을 포함하는 UBR 박스 도메인 기질 결합 억제용 조성물.
  3. 제1항에 있어서,
    상기 A1은 CH3 또는 NH2인 화합물 또는 이의 제약상 허용되는 염을 포함하는 UBR 관련 질환 치료용 약학적 조성물.
  4. 제3항에 있어서,
    상기 UBR 관련 질환은 근육 위축증으로 발병되는 근육 손실증(Becker, Congennital, Duchenne, Distal, Emery-Dreifuss, Facioscapulohumeral, Limb-girdle, myotonic, ocuophargyngeal)(muscular dystrophy), 근육감소증(sarcopenia)이나 암 악액질(cancer cachexia)을 포함한 근육 손실이나 분해로 매개되는 질병(muscle wasting disease)들과 더불어 과다한 단백질 분해로 발병되는 지방육종(liposarcoma), 낭포성섬유증(cystic fibrosis), 요한슨-블리자드 질병(Johanson-Blizzard syndrome), 폐쇄성 요로질환(urethral obstruction sequence), 자가면역 췌장염(autoimmune pancreatitis) 또는 어셔 질병(Usher syndrome)과 같은 UBR box와 UBR 단백질에 관련된 질환으로 공지된 질환들을 포함하는 치료용 약학적 조성물.
PCT/KR2021/005335 2020-04-27 2021-04-27 Ubr 박스 도메인 리간드로의 화합물 Ceased WO2021221444A1 (ko)

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CN202180044509.4A CN115768744B (zh) 2020-04-27 2021-04-27 作为ubr盒结构域配体的化合物
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023074937A1 (ko) * 2021-10-27 2023-05-04 주식회사 오토텍바이오 Ubr 박스 도메인 리간드로의 화합물

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007056464A1 (en) * 2005-11-09 2007-05-18 Proteolix, Inc. Compounds for enzyme inhibition
KR20150080706A (ko) * 2014-01-02 2015-07-10 서울대학교산학협력단 N-말단 법칙 경로의 저해용 조성물 및 방법
JP2015526433A (ja) * 2012-08-14 2015-09-10 フルオリノブ ファーマ インコーポレイテッド フッ素化エポキシケトン系化合物、およびプロテアソーム阻害物質としてのその使用
US20160185785A1 (en) * 2014-12-30 2016-06-30 Forma Therapeutics, Inc. Pyrrolo and pyrazolopyrimidines as ubiquitin-specific protease 7 inhibitors

Family Cites Families (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5464738A (en) 1995-01-06 1995-11-07 Minnesota Mining And Manufacturing Company Sulfonyl hydrazide developers for photothermographic and thermographic elements
DE60112272T2 (de) * 2000-04-27 2006-05-24 Astellas Pharma Inc. Imidazopyridin-derivate
CN1339485A (zh) * 2000-08-23 2002-03-13 上海博德基因开发有限公司 一种新的多肽——人泛素蛋白相关蛋白46.64和编码这种多肽的多核苷酸
MXPA02010231A (es) * 2001-11-27 2004-12-13 Warner Lambert Co Inhibidores aminotransferasa dependientes de aminoacidos de cadena ramificada y su uso en el tratamiento de las enfermedades neurodegenerativas.
CA2892042C (en) * 2012-11-29 2022-06-14 Chemocentryx, Inc. Cxcr7 antagonists
GB201510019D0 (en) * 2015-06-09 2015-07-22 Cancer Therapeutics Crc Pty Ltd Compounds
KR101890421B1 (ko) * 2016-10-06 2018-08-21 한국유나이티드제약 주식회사 Plk1 저해 활성을 갖는 신규 하이드라진 유도체 및 이의 용도
WO2018102419A1 (en) * 2016-11-29 2018-06-07 Epizyme, Inc. Compounds containing a sulfonic group as kat inhibitors
JP2020503363A (ja) * 2017-01-06 2020-01-30 ビヨンドスプリング ファーマシューティカルズ，インコーポレイテッド チューブリン結合化合物およびその治療的使用
PT3652178T (pt) * 2017-07-14 2024-03-18 H Hoffnabb La Roche Ag Compostos bicíclicos de cetona e métodos de utilização dos mesmos
WO2019084030A1 (en) * 2017-10-24 2019-05-02 Genentech, Inc. (4-HYDROXYPYRROLIDIN-2-YL) -HYDROXAMATE COMPOUNDS AND METHODS OF USE
CN112457255B (zh) * 2017-11-08 2022-08-19 北京嘉林药业股份有限公司 化合物及其治疗癌症的用途
BR112020010628A2 (pt) * 2017-11-29 2020-11-10 Epizyme, Inc. composto, composição, método de inibição da atividade de histona acetiltransferases, método para inibir seletivamente a atividade de acetiltransferase, método de tratamento de uma doença ou distúrbio, método de tratamento de um tumor e composto ou composição para uso

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007056464A1 (en) * 2005-11-09 2007-05-18 Proteolix, Inc. Compounds for enzyme inhibition
JP2015526433A (ja) * 2012-08-14 2015-09-10 フルオリノブ ファーマ インコーポレイテッド フッ素化エポキシケトン系化合物、およびプロテアソーム阻害物質としてのその使用
KR20150080706A (ko) * 2014-01-02 2015-07-10 서울대학교산학협력단 N-말단 법칙 경로의 저해용 조성물 및 방법
US20160185785A1 (en) * 2014-12-30 2016-06-30 Forma Therapeutics, Inc. Pyrrolo and pyrazolopyrimidines as ubiquitin-specific protease 7 inhibitors

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
DATABASE REGISTRY CHEMICAL ABSTRACTS SERVICE, COLUMBUS, OHIO, US; 9 July 2015 (2015-07-09), "Pentanamide, N-(5-acetyl-2-chlorophenyl)-5-methoxy- (", XP055865435, retrieved from STN Database accession no. RN: 1797947-97-7 *
See also references of EP4144720A4 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023074937A1 (ko) * 2021-10-27 2023-05-04 주식회사 오토텍바이오 Ubr 박스 도메인 리간드로의 화합물

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