WO2023018155A1 - Shp2 단백질 분해활성을 갖는 화합물 및 이들의 의약 용도 - Google Patents

Shp2 단백질 분해활성을 갖는 화합물 및 이들의 의약 용도 Download PDF

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WO2023018155A1
WO2023018155A1 PCT/KR2022/011797 KR2022011797W WO2023018155A1 WO 2023018155 A1 WO2023018155 A1 WO 2023018155A1 KR 2022011797 W KR2022011797 W KR 2022011797W WO 2023018155 A1 WO2023018155 A1 WO 2023018155A1
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carboxamido
alkyl
methyl
methylthiazol
dimethylbutanoyl
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French (fr)
Inventor
이송희
류제호
이지혜
안정민
배온누리
한정화
임예슬
김지영
이수현
박지윤
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Ubix Therapeutics Inc
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Ubix Therapeutics Inc
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Priority to EP22856152.8A priority Critical patent/EP4385991A4/en
Priority to US18/682,471 priority patent/US20250230154A1/en
Priority to JP2024506263A priority patent/JP2024528949A/ja
Priority to KR1020247007800A priority patent/KR20240050360A/ko
Priority to CN202280055537.0A priority patent/CN117881678A/zh
Publication of WO2023018155A1 publication Critical patent/WO2023018155A1/ko
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
    • 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
    • A61K31/4965—Non-condensed pyrazines
    • A61K31/497—Non-condensed pyrazines containing further heterocyclic rings
    • 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
    • A61K31/4995—Pyrazines or piperazines forming part of bridged ring systems
    • 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
    • A61K31/50—Pyridazines; Hydrogenated pyridazines
    • A61K31/501—Pyridazines; Hydrogenated pyridazines not condensed and containing further heterocyclic rings
    • 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
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—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
    • 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
    • C07D471/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/08—Bridged systems

Definitions

  • the present disclosure relates to a group of compounds having SHP2 proteolytic activity.
  • the present disclosure relates to a group of compounds having a specific structure and having an excellent activity of degrading SHP2 protein.
  • the present disclosure also relates to useful methods of treating diseases associated with the SHP2 protein using such compounds. That is, the present disclosure relates to the pharmaceutical use of the compounds according to the present disclosure for treating or preventing SHP2 protein related diseases.
  • Src homology 2 domain-containing phosphatase is a protein tyrosine phosphatase. Mutations in SHP2 are prevalent in Noonan syndrome and LEOPARD syndrome. Activated mutations of SHP2 have also been identified in pediatric myelomonocytic leukemia, myelodysplastic syndrome, B-cell acute lymphoblastic leukemia and acute myelogenous leukemia.
  • Somatic activating mutations in SHP2 are associated with several types of solid tumors, including lung adenocarcinoma, colon cancer, neuroblastoma, glioblastoma, melanoma, hepatocellular carcinoma, prostate cancer and breast cancer (Bentires-Alj et al., Cancer. Res. 2004, 64, 8816-8820).
  • SHP2 is involved in various signaling processes such as the RAS-ERK, JAK-STAT, PI3K-AKT, NF- ⁇ B and mTOR pathways within cancer cells.
  • SHP2 acts as an upstream positive regulator to promote the RAS-RAF-MEK-ERK kinase cascade signaling.
  • SHP2 inhibition inhibits the dephosphorylation of ERK and the oncogenic function of the RAS-RAF-MEK-ERK pathway, resulting in cell growth inhibition and apoptosis in cancer cells.
  • SHP2 also participates in programmed cell death pathways (PD-1/PD-L1) and contributes to immune evasion by inhibiting T cell activation.
  • PD-1/PD-L1 programmed cell death pathways
  • SHP2 is a very attractive cancer therapeutic target.
  • the SHP2 inhibitor SHP099 has been shown to selectively block SHP2 phosphatase activity and inhibit cancer cell growth in vitro and tumor growth in a xenograft mouse model (Chen et al., Nature 2016, 535, 148-152; Garcia Fortanet et al., J. Med. Chem. 2016, 59, 7773-7782).
  • Allosteric SHP2 inhibitors have been shown to be effective in a preclinical model of Kirsten rat sarcoma (KRAS)-mutant human cancer. Deletion of the SHP2 protein may also provide an alternative and possibly even more effective strategy for inhibiting SHP2 activity. For example, it has been demonstrated that SHP2 inactivation by CRISPR-Cas9 induces senescence and inhibits tumor growth in a xenograft model of KRAS-mutant tumors. These findings provide evidence that ablation of the SHP2 protein in tumor cells can be an effective therapeutic strategy for human cancers, particularly those with KRAS-mutations.
  • the problems to be solved by the present invention are compounds having Src homology 2 domain-containing phosphatase (SHP2) degradation activity, pharmaceutical compositions containing them as active ingredients, and their SHP2-related diseases ( Preferably, it is to provide a pharmaceutical use for cancer or tumor) treatment or prevention.
  • SHP2 Src homology 2 domain-containing phosphatase
  • Another problem to be solved by the present invention is that the compound according to the present invention, characterized in that it degrades SHP2 and consequently lowers SHP2 activity, is administered to patients in need of treatment, improvement or prevention of SHP2-related diseases, characterized in that SHP2 It is to provide a method for treating or improving a related disease (preferably, cancer or tumor).
  • a related disease preferably, cancer or tumor.
  • the present invention provides a compound of Formula 1 below or a pharmaceutically acceptable salt thereof.
  • R 1a , R 1b , R 1c , and R 1d are each independently H or C 1-6 alkyl (preferably methyl),
  • R 2 is H, C 1-6 alkyl, or -NH 2 ;
  • R 3 is H, C 1-6 alkyl, or C 1-6 hydroxyalkyl
  • R 4 is H, C 1-6 alkyl, halogen (preferably Cl), or C 1-6 haloalkyl;
  • R 5 is H, C 1-6 alkyl, C 1-6 alkynyl, halogen, -CN, or heteroaryl (preferably thiazole), wherein heteroaryl is optionally one or more hydrogens in the ring are C 1-6 alkyl ( preferably methyl), halogen, or C 1-3 haloalkyl,
  • R 6 is -NHC(O)R 7 or heteroaryl, wherein heteroaryl optionally has one or more hydrogens in the ring substituted with C 1-6 alkyl, halogen, or C 1-3 haloalkyl;
  • R 7 is C 1-6 alkyl (preferably methyl), C 1-6 haloalkyl, C 3-10 cycloalkyl (preferably cyclopropyl), or a heterocycle, wherein the cycloalkyl and heterocycle are independently of each other While optionally, one or more hydrogens in the ring are substituted with C 1-6 alkyl, halogen (preferably F), or -CN,
  • X is CH or N
  • n 1 and n 2 are each independently 0, 1, or 2;
  • n 0 or 1
  • a 1 , A 2 , and A 3 are, independently of each other, a direct bond, -O-, -N(R 9 )- -C(O)-, -C(O)N(R 9 )-, or -N( R 9 )C(O)-;
  • heterocycle preferably azetidine, pyrrolidine, piperidine, piperazine, azaspiro[3.5]nonane, diazabicyclo[3.2.1 ]octane
  • aryl, or heteroaryl preferably pyridine,
  • R 8 and R 8 ' are independently H or C 1-6 alkyl, or R 8 and R 8 'are linked to each other to form C 3-10 cycloalkyl (preferably cyclopropyl);
  • R 9 is hydrogen or C 1-6 alkyl (preferably methyl);
  • q 1 , q 2 , q 3 , q 4 , and q 5 are independently integers from 0 to 10;
  • the compound according to the present invention is a protac of 'SHP2 binding ligand-linker-E3 ligase ligand' ('left moiety based on L-linker (L) -right moiety based on L' in Formula 1), PROTAC (Proteolysis- targeting chimera) compound.
  • the present inventors have introduced various E3 ligase ligands such as CRBN ligand, VHL ligand, and IAP ligand to the SHP2 binding moiety of Formula 1 of the present invention, and among these ligands, only the VHL ligand such as Formula 1 of the present invention has the effect. was excellent.
  • the present invention is based on these unique and surprising findings, the present invention is not limited to these theoretical mechanisms.
  • the present inventors evaluated various substituents, and in the case of a structure such as Formula 1 of the present disclosure, in particular, in the case of a preferred structure described later, excellent compounds in various aspects including SHP2 decomposition activity could be obtained. That is, the present invention provides a new compound with excellent SHP2 decomposition activity, (metabolism) stability, etc., and excellent physicochemical properties (cLogP value, water solubility, cell membrane permeability) as a medicinal ingredient.
  • substituent may be (1) unsubstituted or (2) substituted with one or more of the defined substituents. If a substitutable position is unsubstituted, the default substituent is hydrogen.
  • alkyl refers to a saturated straight-chain or branched non-cyclic hydrocarbon having 1 to 10 carbon atoms (unless the number of carbon atoms is specifically limited). "Lower alkyl” means a straight chain or branched alkyl having 1 to 4 carbon atoms.
  • saturated straight-chain alkyls are -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl, -n-hexyl, -n-heptyl, -n-octyl, -n-nonyl and -n- decyl, while saturated branched alkyl is -isopropyl, -sec-butyl, -isobutyl, -tert-butyl, isopentyl, 2-methylhexyl, 3-methylbutyl, 2-methylpentyl, 3- Methylpentyl, 4-methylpentyl, 2-methylhexyl, 3-methylhexyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2-methylhexyl, 3-methylhexyl, 4-methylpentyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-
  • alkynyl refers to a straight-chain or branched non-cyclic hydrocarbon having 2 to 10 carbon atoms and containing at least one carbon-carbon triple bond (if the number of carbon atoms is not particularly limited). do.
  • Representative straight-chain or branched (C 2 -C 10 )alkynyls are -acetyrenyl, -propynyl, -1-butynyl, -2-butynyl, -1-pentynyl, -2-pentynyl, - 3-methyl-1-butynyl, -4-pentynyl, -1-hexynyl, -2-hexynyl, -5-hexynyl, -1-heptynyl, -2-heptynyl, -6-heptynyl , -1-octynyl, -2-octynyl, -7-octynyl, -1-nonynyl, -2-nonynyl, -8-nonynyl, -1-decynyl, -2-decynyl, and -9-decyny
  • C 1-6 alkyl means an alkyl having 1 to 6 carbon atoms.
  • haloalkyl refers to an alkyl group in which each of one or more hydrogen atoms is replaced with a halogen atom.
  • haloalkyl is -CF 3 , -CHF 2 , -CH 2 F, -CBr 3 , -CHBr 2 , -CH 2 Br, -CC1 3 , -CHC1 2 , -CH 2 CI, -CI 3 , -CHI 2 , -CH 2 I, -CH 2 -CF 3 , -CH 2 -CHF 2 , -CH 2 -CH 2 F, -CH 2 -CBr 3 , -CH 2 -CHBr 2 , -CH 2 -CH 2 Br, -CH 2 -CC1 3 , -CH 2 -CHC1 2 , -CH 2 -CH 2 CI, -CH 2 -CI 3 , -CH 2 -CHI 2 ,
  • hydroxyalkyl refers to a linear or branched C 1-10 alkyl substituted with one or more hydroxy groups, examples of hydroxyalkyl groups include hydroxymethyl, hydroxyethyl, hydroxypropyl and hydroxybutyl, but is not limited thereto.
  • cycloalkyl means a monocyclic or polycyclic saturated ring having carbon and hydrogen atoms and not having carbon-carbon multiple bonds.
  • monocyclic rings include, but are not limited to, (C 3 -C 7 )cycloalkyl (eg, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl).
  • polycyclic rings examples include fused bicyclic rings such as octahydropentalene, decahydronaphthalene, and the like; spiro rings such as spiro[3.3]heptane, spiro[3.4]octane, spiro[3.5]nonane, spiro[4.4]nonane, spiro[4.5]decane, spiro[5.5]undecane and the like; and bridged bicyclic rings such as bicyclo[2.1.1]hexane, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, and the like. Cycloalkyl groups may be optionally substituted. In one embodiment, a cycloalkyl group is a monocyclic ring (ring). In one preferred aspect of the invention, cycloalkyl is cyclopropyl.
  • a “heterocycle” or “heterocycloalkyl” is a saturated 5- to 7-membered monocyclic ring containing 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur. , or a 7- to 12-membered bicyclic ring (ring) in which nitrogen and sulfur heteroatoms can be selectively oxidized and nitrogen heteroatoms can be selectively quaternized.
  • heterocycles include oxiran, oxetan, tetrahydrofuran, tetrahydropyran, 1,4-dioxane, aziridine, azetidine, pyrrolidine, piperidine, piperazine, pyrrolidinone, hydantoine, valerolactam, thiirane ), thietane, tetrahydrothiophene, tetrahydrothiopyra, morpholine, tetrahydropyridine, tetrahydropyrimidine, and the like.
  • Heterocycles include bicyclic rings in which a portion of the heterocycle is fused to a benzene or cyclopenta-1,3-diene ring. Heterocycles may be attached by heteroatoms or carbon atoms. Heterocycles also include fused bicyclic rings, spiro rings and bridged bicyclic rings in which one or more carbon atoms of the aforementioned polycyclic rings are replaced with nitrogen, oxygen or sulfur atoms. ring is included.
  • heterobicyclics such as octahydrocyclopenta[c]pyrrole, octahydropyrrolo[3,4-c]pyrrole, decahydroisoquinoline, decahydro-2,6-naphthyridine, etc.
  • the heterocycle is any one selected from azetidine, diazetidine, piperidine, piperazine, azaspiro[3.5]nonane, diazabicyclo[3.2.1]octane, and the like.
  • aryl refers to a carbocyclic aromatic group containing 5 to 10 ring atoms. Representative examples include phenyl, tolyl, xylyl, naphthyl, tetrahydronaphthyl, anthracenyl, fluorenyl, indenyl, azulenyl, and the like. Including, but not limited to. Carbocyclic aromatic groups may be optionally substituted.
  • heteroaryl has at least one heteroatom selected from the group consisting of nitrogen, oxygen, and sulfur, and contains from 5 to 10 carbon atoms containing at least one carbon atom, including mono- and bicyclic ring systems. It is an aromatic heterocycle ring of a member.
  • heteroaryls include furan, 4H-pyran, pyrrole, imidazole, pyrazole, triazole, tetrazole, pyridine, pyrimidine, pyridazine, pyrazine, triazine, thiophene, ozaxole, isoxazole, thiazole, isothiazole, oxadiazole, benzofuran, benzothiophene, quinoline, indole, These include benzoxazole, benzimidazole, benzothiazole, cinnoline, phthalazine, quinazoline, and 1H-azepine.
  • heteroaryl is thiazole, pyridine, or pyrazine.
  • a preferred aspect of the present invention is in Formula 1 above,
  • R 1a , R 1b , R 1c , and R 1d are each independently H or C 1-6 alkyl (preferably methyl),
  • R 2 is H, C 1-6 alkyl, or -NH 2 ;
  • R 3 is H, C 1-6 alkyl, or C 1-6 hydroxyalkyl
  • R 4 is H, C 1-6 alkyl, or halogen (preferably Cl);
  • R 5 is heteroaryl (preferably thiazole), wherein heteroaryl optionally has one or more hydrogens in the ring substituted with C 1-6 alkyl (preferably methyl) or halogen,
  • R 6 is -NHC(O)R 7 ;
  • R 7 is C 1-6 alkyl (preferably methyl) or C 3-10 cycloalkyl (preferably cyclopropyl), wherein cycloalkyl optionally one or more hydrogens in the ring is C 1-6 alkyl, halogen ( preferably F), or -CN,
  • X is CH or N
  • n 1 and n 2 are each independently 0 or 1;
  • n 0 or 1
  • a 1 , A 2 , and A 3 are, independently of each other, a direct bond, -O-, -N(R 9 )- -C(O)-, -C(O)N(R 9 )-, or -N( R 9 )C(O)-;
  • B 1 and B 2 independently represent a direct bond, a heterocycle (preferably azetidine, pyrrolidine, piperidine, piperazine, azaspiro[3.5]nonane, diazabicyclo[3.2.1]octane), or heteroaryl.
  • a heterocycle preferably azetidine, pyrrolidine, piperidine, piperazine, azaspiro[3.5]nonane, diazabicyclo[3.2.1]octane
  • heteroaryl preferably pyridine, pyrimidine, pyridazine, pyrazine
  • R 8 and R 8 ' are independently H or C 1-6 alkyl, or R 8 and R 8 'are linked to each other to form C 3-10 cycloalkyl (preferably cyclopropyl);
  • R 9 is hydrogen or C 1-6 alkyl (preferably methyl);
  • q 1 , q 2 , q 3 , q 4 , and q 5 are independently integers from 0 to 10;
  • a compound or a pharmaceutically acceptable salt thereof is provided.
  • R 1a , R 1b , R 1c , and R 1d are each independently H or C 1-6 alkyl (preferably methyl),
  • R 2 is H or -NH 2 ;
  • R 3 is H
  • R 4 is H or halogen (preferably Cl);
  • R 5 is thiazole, wherein thiazole optionally has one or more hydrogens in the ring substituted with C 1-6 alkyl (preferably methyl);
  • R 6 is -NHC(O)R 7 ;
  • R 7 is C 1-6 alkyl (preferably methyl) or cyclopropyl, wherein cyclopropyl optionally has one or more hydrogens in the ring replaced by halogen (preferably F) or -CN;
  • X is CH
  • n 1 and n 2 are each independently 0 or 1;
  • n 0 or 1
  • a 1 , A 2 , and A 3 are, independently of each other, a direct bond, -O-, -N(R 9 )- -C(O)-, -C(O)N(R 9 )-, or -N( R 9 )C(O)-;
  • B 1 and B 2 are, independently of each other, a direct bond, azetidine, pyrrolidine, piperidine, piperazine, azaspiro[3.5]nonane, diazabicyclo[3.2.1]octane, pyridine, pyrimidine, pyridazine, or pyrazine.
  • B 1 and B 2 is azetidine, pyrrolidine, piperidine, piperazine, azaspiro[3.5]nonane, diazabicyclo[3.2.1]octane, pyridine, pyrimidine, pyridazine, or pyrazine;
  • R 8 and R 8 ' are independently H or C 1-6 alkyl, or R 8 and R 8 'are linked to each other to form C 3-10 cycloalkyl (preferably cyclopropyl);
  • R 9 is hydrogen or C 1-6 alkyl (preferably methyl);
  • q 1 , q 2 , q 3 , q 4 , and q 5 are independently integers from 0 to 10;
  • a compound or a pharmaceutically acceptable salt thereof is provided.
  • the linker (L) is any one of the following linkers.
  • SHP2 degradation activity is excellent, and is more suitable for various purposes of the present invention.
  • Non-limiting examples of compounds of Formula 1 according to the present disclosure are the compounds prepared in the Examples described below. Each example number corresponds to a compound number. For example, the number of the final compound prepared in Example 10 is compound 10.
  • the compounds in Table 1 below were particularly preferred in terms of various aspects such as SHP2 degradation activity, cancer cell line cytotoxicity, (metabolism) stability, and physicochemical properties, particularly in terms of SHP2 degradation activity.
  • “pharmaceutically acceptable salts” include salts of active compounds prepared with relatively non-toxic acids and bases depending on the specific substituents found in the compounds mentioned herein.
  • base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either pure or in a suitable inert solvent.
  • pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino or magnesium salts or similar salts.
  • acidic addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either pure or in a suitable inert solvent.
  • Examples of pharmaceutically acceptable acid addition salts are acetic acid, propionic acid, isobutyric acid, oxalic acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid ( fumaric), mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric acid, tartaric acid, methanesulfonic, and their analogues.
  • Hydrogen chloride hydrogen bromide, nitric acid, carbonic acid, monohydrogencarbonic, phosphoric, monohydrogenphosphate, dihydrogenphosphate, sulfuric acid, monohydrosulfuric acid, hydrogen iodide or phosphorous acid ( phosphorous acid) and its analogues.
  • salts of amino acids such as alginate and analogs thereof and analogs of organic acids such as glucuronic or galactunoric acids and analogs thereof.
  • Some particular compounds of the present invention have both basic and acidic functionality which allows them to be converted into basic or acidic addition salts.
  • Other examples of salts are well known from the literature known in the art to which this invention pertains.
  • the term "compound of the present invention” is meant to include not only each compound of Formula 1, but also clathrates, hydrates, solvates, or polymorphs thereof.
  • the term “compound of the present invention” is meant to include pharmaceutically acceptable salts of the compounds of the present invention when pharmaceutically acceptable salts thereof are not mentioned.
  • the compounds of the present invention are stereomerically pure compounds (e.g., substantially free of other stereoisomers (e.g., greater than 85% ee, greater than 90% ee, greater than 95% ee, 97% ee or more, or 99% ee or more))).
  • the compound of Formula 1 or a salt thereof according to the present invention is a tautomeric isomer and/or a stereoisomer (eg, geometrical isomer and conformational isomers), the separated isomer thereof and mixtures each are also included within the scope of the compounds of the present invention.
  • the compounds of the present invention or their salts have an asymmetric carbon in their structure, their optically active compounds and racemic mixtures are also included in the scope of the compounds of the present invention.
  • polymorph refers to a solid crystal form of a compound of the present invention or a complex thereof. Different polymorphs of the same compound exhibit different physical, chemical and/or spectral properties. Differences in terms of physical properties include, but are not limited to, stability (e.g. heat or light stability), compressibility and density (important for formulation and product manufacturing), and dissolution rate (which may affect bioavailability). It doesn't. Differences in stability may be due to chemical reactivity changes (e.g. differential oxidation such as faster discoloration when composed of one polymorph than when composed of another polymorph) or mechanical properties (e.g.
  • kinetically Tablet fragments stored as the preferred polymorph may be thermodynamically converted to a more stable polymorph) or both (tablets of one polymorph are more susceptible to degradation at high humidity).
  • Other physical properties of polymorphs can affect their processing. For example, one polymorph may be more likely to form solvates, or may be more difficult to filter or wash, than another polymorph, eg, due to its shape or particle size distribution.
  • solvent compound refers to a compound of the present invention or a pharmaceutically acceptable salt thereof comprising a stoichiometric or non-stoichiometric amount of a solvent bound by non-covalent intermolecular forces.
  • Preferred solvents are volatile, non-toxic, and can be administered in very small amounts to humans.
  • hydrate refers to a compound of the present invention, or a pharmaceutically acceptable salt thereof, containing a stoichiometric or non-stoichiometric amount of water bound by non-covalent intermolecular forces.
  • clathrate refers to a compound of the present invention in the form of a crystal lattice containing spaces (eg, channels) in which guest molecules (eg, solvent or water) are confined. or a salt thereof.
  • the term "purified" means that when isolated, the isolate is at least 90% pure, in one embodiment at least 95% pure, in another embodiment at least 99% pure, and In other embodiments, at least 99.9% pure.
  • the invention further provides a method of treating a disease or condition in a subject having or susceptible to having one or more of the following diseases or conditions by administering to the subject a therapeutically effective amount of one or more such compounds.
  • the treatment is a preventative treatment.
  • the treatment is a palliative treatment.
  • the treatment is a restorative treatment.
  • the compounds for degrading SHP2 of the present invention are useful for various therapeutic or prophylactic applications (eg, cancer). These compounds can be used to lower SHP2 activity by degrading SHP2, and can also be used to treat or prevent aggravation of SHP2-related diseases.
  • the present invention provides a method for degrading intracellular SHP2.
  • the cells are contacted with an effective amount of a compound of the present invention.
  • the cell is within a subject.
  • the method of the present invention comprises administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically or prophylactically effective amount of a compound according to the present invention.
  • the present invention provides a method for degrading SHP2 in cells of a SHP2-associated disease.
  • the present invention can be used to degrade SHP2 in the cells of a subject with a SHP2-related disease described below, thereby lowering SHP2 activity.
  • the invention relates to cancer, particularly leukemia, lymphoma, lung cancer, head and neck cancer, esophageal cancer, stomach cancer, colorectal cancer, pancreatic cancer, liver cancer, breast cancer, ovarian cancer, cervical cancer, bladder cancer, melanoma, neuroblastoma, neuroblastoma It can be used to degrade SHP2 in cells of glioma, sarcoma, etc.
  • the present invention provides a method of treating a SHP2-related disease comprising administering to a subject a therapeutically effective amount of a compound of Formula 1 or a pharmaceutically acceptable salt thereof.
  • Such methods include administering to a subject in need thereof an amount of a compound of the present invention sufficient to degrade SHP2, i.e., a therapeutically effective amount.
  • a compound of the present invention can be administered to the subject in the form of a pharmaceutical composition described herein.
  • the SHP2 related disease is cancer.
  • it may be any one of the cancers listed in Table 2 of International Patent Application Publication WO 2021/236775, but is not limited thereto.
  • the description of Table 2 of WO 2021/236775 is fully incorporated herein by reference.
  • the cancer is a solid tumor.
  • the cancer is a hematological cancer. Exemplary hematological cancers include, but are not limited to, cancers listed in Table 3 of WO 2021/236775. The description of Table 3 of WO 2021/236775 is fully incorporated herein by reference.
  • the hematological cancer is acute lymphocytic leukemia, chronic lymphocytic leukemia (including B-cell chronic lymphocytic leukemia), or acute myeloid leukemia.
  • the cancer is a leukemia, eg, a leukemia selected from acute monocytic leukemia, acute myelogenous leukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia and mixed lineage leukemia (MLL).
  • MMLL mixed lineage leukemia
  • the cancer is NUT-midline carcinoma.
  • the cancer is multiple myeloma.
  • the cancer is lung cancer, such as small cell lung cancer (SCLC).
  • SCLC small cell lung cancer
  • the cancer is a neuroblastoma.
  • the cancer is Burkitt's lymphoma. In another embodiment, the cancer is cervical cancer. In another embodiment, the cancer is head and neck cancer. In another embodiment, the cancer is esophageal cancer. In another embodiment, the cancer is gastric cancer. In another embodiment, the cancer is pancreatic cancer. In another embodiment, the cancer is liver cancer. In another embodiment the cancer is melanoma. In another embodiment, the cancer is ovarian cancer. In another embodiment, the cancer is colorectal cancer. In another embodiment, the cancer is prostate cancer. In another aspect, the cancer is breast cancer.
  • the cancer according to the present invention is acute monocytic leukemia, acute myeloid leukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia, mixed lineage leukemia, lymphoma, NUT-midline carcinoma, multiple myeloma, small cell lung cancer, non-small cell lung cancer , neuroblastoma, Burkitt's lymphoma, cervical cancer, head and neck cancer, esophageal cancer, gastric cancer, pancreatic cancer, liver cancer, melanoma, ovarian cancer, colon cancer, prostate cancer, breast cancer, bladder cancer, glioma, sarcoma, esophageal squamous cell carcinoma, and papillary thyroid carcinoma is selected from the group consisting of
  • the present invention provides a pharmaceutical use for treating or preventing the above diseases of the compound of Formula 1 or a pharmaceutically acceptable salt thereof.
  • Suitable subjects to be treated according to the present invention include mammalian subjects.
  • Mammals according to the present invention include, but are not limited to, humans, canines, felines, bovines, caprines, equines, ovines, and pigs. (porcine), rodents (rodents), lagomorphs (lagomorphs), primates (primates) and the like, including mammals in utero ( in utero ).
  • a suitable subject to be treated according to the present invention is a human.
  • a compound of the present invention is generally administered in a therapeutically effective amount.
  • an “effective amount” refers to an amount of a compound of the present invention sufficient to slow or minimize the progression of a SHP2-related disease or to provide therapeutic benefit in the treatment or management of a SHP2-related disease. “Effective amount” also refers to an amount sufficient to inhibit or reduce SHP2 activity, either in vitro or in vivo .
  • the compounds of the present invention can be administered by any suitable route, in the form of a pharmaceutical composition suitable for such route, and in a dosage effective for the intended treatment.
  • An effective dosage is generally from about 0.001 to about 100 mg/kg of body weight/day, preferably from about 0.01 to about 50 mg/kg/day, in single or divided administration. Dosage levels below the lower end of this range may be suitable depending on the age, species, and disease or condition being treated. In other cases, still larger doses can be used without detrimental side effects. Larger doses may be divided into several smaller doses for administration throughout the day.
  • the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of Formula 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or additive.
  • the use of the pharmaceutical composition is for the treatment or prevention of SHP2-related diseases, preferably cancer, which will be described later.
  • pharmaceutically acceptable means suitable for use as a pharmaceutical preparation, generally considered safe for such use, and officially approved for such use by a national regulatory agency or approved by the Korean Pharmacopoeia or the United States Means in the pharmacopeia list.
  • the compound described herein, or a pharmaceutically acceptable salt thereof can be administered as follows.
  • the compound of the present invention can be administered orally, and oral is a concept including swallowing.
  • Oral administration allows the compounds of the present invention to enter the gastrointestinal tract or be directly absorbed from the mouth into the bloodstream, eg, by buccal or sublingual administration.
  • compositions suitable for oral administration may be in solid, liquid, gel, or powder form, and may have formulations such as tablets, lozenges, capsules, granules, powders, and the like. .
  • compositions for oral administration may optionally be enteric coated and exhibit delayed or sustained release through the enteric coating. That is, the composition for oral administration according to the present invention may be a formulation having an immediate or modified release pattern.
  • Liquid formulations may include solutions, syrups and suspensions, and such liquid compositions may be contained in soft or hard capsules.
  • Such formulations may include a pharmaceutically acceptable carrier such as water, ethanol, polyethylene glycol, cellulose, or oil.
  • the formulation may also contain one or more emulsifying and/or suspending agents.
  • the amount of active ingredient drug may be present from about 0.05% to about 95% by weight, more typically from about 2% to about 50% by weight of the total weight of the tablet.
  • Tablets may also contain a disintegrant comprising from about 0.5% to about 35% by weight, more usually from about 2% to about 25% by weight of the dosage form.
  • disintegrants include, but are not limited to, lactose, starch, sodium starch glycolate, crospovidone, croscarmellose sodium, maltodextrin, or mixtures thereof.
  • Suitable glidants included for making into tablets may be present in amounts from about 0.1% to about 5% by weight, and include talc, silicon dioxide, stearic acid, calcium, zinc or magnesium stearate, sodium stearyl fumarate, and the like. It can be used as a lubricant, but the present invention is not limited to these types of additives.
  • Gelatin polyethylene glycol, sugar, gum, starch, polyvinylpyrrolidone, hydroxypropylcellulose, hydroxypropylmethylcellulose, etc. may be used as a binder for preparing tablets.
  • Mannitol, xylitol, lactose, dextrose, sucrose, sorbitol, starch, microcrystalline cellulose, etc. may be used as suitable diluents for preparing tablets, but the present invention is not limited to these types of additives. .
  • the solubilizing agent that may be included in the tablet may be used in an amount of about 0.1% to about 3% by weight based on the total weight of the tablet, for example, polysorbate, sodium lauryl sulfate, sodium dodecyl sulfate, propylene carbonate, Diethylene glycol monoethyl ether, dimethylisosorbide, polyoxyethylene glycolated natural or hydrogenated castor oil, HCOR TM (Nikkol), oleyl ester, Gelucire TM , caprylic/caprylic mono/ Diglyceride, sorbitan fatty acid ester, Solutol HS TM and the like can be used in the pharmaceutical composition according to the present invention, but the present invention is not limited to the specific types of these solubilizing agents.
  • Compounds of the present invention may be administered directly into the bloodstream, muscle, or intestine.
  • Suitable methods for parenteral administration include intravenous, intramuscular, subcutaneous intraarterial, intraperitoneal, intrathecal, intracranial injection, and the like.
  • Suitable devices for parenteral administration include injectors (including needle and needleless syringes) and infusion methods.
  • compositions for parenteral administration may be formulations with an immediate or modified release pattern, and the modified release pattern may be a delayed or sustained release pattern.
  • parenteral formulations are liquid compositions, and these liquid compositions are aqueous solutions containing the active ingredient, salt, buffer, tonicity agent and the like according to the present invention.
  • Parenteral formulations may also be prepared in dried form (eg lyophilized) or as sterile non-aqueous solutions. These formulations may be used with a suitable vehicle such as sterile water. Solubility-enhancing agents may also be used in the preparation of parenteral solutions.
  • the compounds of the present invention may be administered topically to the skin or transdermally.
  • Formulations for this topical administration include lotions, solutions, creams, gels, hydrogels, ointments, foams, implants, patches, and the like.
  • Pharmaceutically acceptable carriers for topical formulations may include water, alcohol, mineral oil, glycerin, polyethylene glycol, and the like. Topical administration can also be performed by electroporation, iontophoresis, phonophoresis, and the like.
  • compositions for topical administration may be formulations with an immediate or modified release pattern, and the modified release pattern may be a delayed or sustained release pattern.
  • the present disclosure provides compounds capable of exhibiting various pharmacological activities due to their excellent SHP2 degradation activity, pharmaceutical compositions containing them as active ingredients, their medicinal uses (particularly, cancer or tumor), and their administration to subjects in need of treatment or prevention. It provides a treatment method that includes.
  • the compounds according to the present invention or pharmaceutically acceptable salts thereof are excellent in various aspects such as SHP2 degrading activity, (metabolic) stability, and physicochemical properties, and in particular, have much better SHP2 degrading activity compared to compounds having similar structures.
  • Step 4 Synthesis of tert-butyl (1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)carbamate
  • Step 1 Synthesis of (2S,4R)-methyl 1-((S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxylate
  • reaction solution was washed with a saturated aqueous solution of NH 4 Cl (300 ml) and a saturated aqueous solution of NaHCO 3 (300 ml), and then extracted with EtOAc (150 ml x 2).
  • EtOAc 150 ml x 2
  • the organic layer was washed with brine (150 ml x 2) and dried over anhydrous sodium sulfate. Thereafter, the residue obtained by filtration and concentration under reduced pressure was subjected to MPLC (0-5% MeOH/DCM) to obtain 11 g (71%) of an ivory solid.
  • Step 2 Synthesis of (2S,4R)-1-((S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxylic acid
  • Step 5 tert-butyl ((S)-1-((2S,4R)-4-hydroxy-2-((2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1 Synthesis of -yl)-3,3-dimethyl-1-oxobutan-2-yl)carbamate
  • Step 7 (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(2-hydroxy-4-(4 Synthesis of -methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide
  • Step 2 (9H-fluoren-9-yl)methyl 4-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2- Synthesis of yl)thio)-2-chlorophenyl)carbamoyl)piperidine-1-carboxylate
  • Step 3 tert-butyl ((1-(5-((2-chloro-3-(piperidine-4-carboxamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate synthesis of
  • Step 4 tert-butyl ((1-(5-((2-chloro-3-(1-(3-(2-(((2S,4R)-1-((S)-2-(1- fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)propyl)piperidine-4-carboxamido)phenyl)thio) Synthesis of pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate
  • Step 5 N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-1-(3-(2-( ((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol Synthesis of -5-yl)phenoxy)propyl)piperidine-4-carboxamide
  • Example 6 was synthesized in the same manner as in Example 1, using 2-(piperidin-4-yl)acetic acid instead of piperidine-4-carboxylic acid.
  • Example 14 was synthesized in the same manner as in Example 1, using 3-(piperidin-4-yl)propanoic acid instead of piperidine-4-carboxylic acid.
  • Example 20 ((2S,4R)-N-(2-(3-(4-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin- 2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)-4-hydroxypiperidin-1-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S) -2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
  • Step 1 Synthesis of 2-(1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-hydroxypiperidin-4-yl)acetic acid
  • Step 2 (9H-fluoren-9-yl)methyl 4-(2-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin Synthesis of -2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)-4-hydroxypiperidine-1-carboxylate
  • Step 3 tert-butyl ((1-(5-((2-chloro-3-(2-(4-hydroxypiperidin-4-yl)acetamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin- Synthesis of 4-yl)methyl)carbamate
  • Step 4 tert-butyl ((1-(5-((2-chloro-3-(2-(1-(3-(2-(((2S,4R)-1-((S)-2- (1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)propyl)-4-hydroxypiperidin-4- Synthesis of yl)acetamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate
  • Step 5 ((2S,4R)-N-(2-(3-(4-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2 -yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)-4-hydroxypiperidin-1-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)- Synthesis of 2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
  • Example 21 N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-1-(4-(2- (((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4- methylthiazol-5-yl)phenoxy)butanoyl)piperidine-4-carboxamide
  • Step 1 ethyl 4-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido Synthesis of )methyl)-5-(4-methylthiazol-5-yl)phenoxy)butanoate
  • Step 2 4-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido) Synthesis of methyl)-5-(4-methylthiazol-5-yl)phenoxy)butanoic acid
  • Step 3 tert-butyl ((1-(5-((2-chloro-3-(1-(4-(2-(((2S,4R)-1-((S)-2-(1- fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)butanoyl)piperidine-4-carboxamido)phenyl)thio) Synthesis of pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate
  • Step 4 N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-1-(4-(2-( ((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol Synthesis of -5-yl)phenoxy)butanoyl)piperidine-4-carboxamide
  • Ethyl 2-bromoacetate was used instead of ethyl 4-bromobutyrate, tert-butyl ((1-(5-((2-chloro-3-(piperidine-4-carboxamido)phenyl)thio)pyrazin-2-yl)-4 tert-butyl ((1-(5-((2-chloro-3-(2-(piperidin-4-yl)acetamido)phenyl)thio)pyrazin-2-yl instead of -methylpiperidin-4-yl)methyl)carbamate
  • Example 22 was synthesized in the same manner as in Example 21 using )-4-methylpiperidin-4-yl)methyl)carbamate.
  • Example 26 was synthesized in the same manner as in Example 21 using )-4-methylpiperidin-4-yl)methyl)carbamate.
  • Step 1 ethyl 2-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl) Synthesis of amino)-2-oxoacetate
  • Step 2 2-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino )-2-oxoacetic acid synthesis
  • Step 3 Synthesis of tert-butyl 4-(2-hydroxyethyl)piperidine-1-carboxylate
  • Step 4 Synthesis of tert-butyl 4-(2-(tosyloxy)ethyl)piperidine-1-carboxylate
  • Step 5 tert-butyl 4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4- Synthesis of hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperidine-1-carboxylate
  • Step 6 (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5 Synthesis of -yl)-2-(2-(piperidin-4-yl)ethoxy)benzyl)pyrrolidine-2-carboxamide
  • Step 7 tert-butyl ((1-(5-((2-chloro-3-(2-(4-(2-(2-(((2S,4R)-1-((S)-2- (1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperidin-1-yl)- Synthesis of 2-oxoacetamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate
  • Step 8 (2S,4R)-N-(2-(2-(1-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2- yl)thio)-2-chlorophenyl)amino)-2-oxoacetyl)piperidin-4-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1 Synthesis of -fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
  • Example 28 was synthesized in the same manner as in Example 27, using 3-(piperidin-4-yl)propan-1-ol instead of 2-(piperidin-4-yl)ethan-1-ol.
  • tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (intermediate 3) instead of tert-butyl (1 -(6-amino-5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)carbamate (intermediate 2) was used, and ethyl 3-
  • Example 30 was synthesized in the same manner as in Example 27, using chloro-3-oxopropanoate and piperidin-4-ylmethanol instead of 2-(piperidin-4-yl)ethan-1-ol.
  • Example 31 was synthesized in the same manner as in Example 27, using ethyl 3-chloro-3-oxopropanoate instead of ethyl chloroglyoxylate and piperidin-4-ylmethanol instead of 2-(piperidin-4-yl)ethan-1-ol. was synthesized.
  • Example 32 was synthesized in the same manner as in Example 27, using ethyl 3-chloro-3-oxopropanoate and piperidin-4-ylmethanol instead of 2-(piperidin-4-yl)ethan-1-ol. .
  • Example 34 was synthesized in the same manner as in Example 27 using -3-oxopropanoate.
  • Example 36 was synthesized in the same manner as in Example 27, using ethyl 3-chloro-3-oxopropanoate instead of ethyl chloroglyoxylate.
  • Example 37 was synthesized in the same manner as in Example 27 using ethyl 3-chloro-3-oxopropanoate.
  • tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (intermediate 3) instead of tert-butyl (( Use 1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (intermediate 1), and use ethyl 3-chloro instead of ethyl chloroglyoxylate -3-oxopropanoate was used, and 3-(piperidin-4-yl)propan-1-ol was used instead of 2-(piperidin-4-yl)ethan-1-ol, and the synthesis was carried out in the same manner as in Example 27.
  • Example 39 was synthesized.
  • Example 27 using ethyl 3-chloro-3-oxopropanoate instead of ethyl chloroglyoxylate and using 3-(piperidin-4-yl)propan-1-ol instead of 2-(piperidin-4-yl)ethan-1-ol
  • Example 41 was synthesized in the same manner as the synthesis method of.
  • Example 43 was synthesized in the same manner as in Example 27 using -(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (Intermediate 9).
  • Example 44 was synthesized in the same manner as in Example 27, using methyl 4-chloro-4-oxobutanoate instead of ethyl chloroglyoxylate and piperidin-4-ylmethanol instead of 2-(piperidin-4-yl)ethan-1-ol. was synthesized.
  • Example 45 was synthesized in the same manner as in Example 27, using methyl 4-chloro-4-oxobutanoate instead of ethyl chloroglyoxylate.
  • Example 46 was synthesized.
  • Example 47 was synthesized in the same manner as in Example 27 using methyl 1-(chlorocarbonyl)cyclopropane-1-carboxylate instead of ethyl chloroglyoxylate.
  • Example 48 was synthesized in the same manner as in Example 27.
  • Example 49 was synthesized in the same manner as in Example 27 using methyl 3-chloro-2,2-dimethyl-3-oxopropanoate instead of ethyl chloroglyoxylate.
  • Step 1 ethyl 3-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl) Synthesis of amino)-3-oxopropanoate
  • Step 2 3-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino )-3-oxopropanoic acid synthesis
  • Step 3 Synthesis of tert-butyl 2-(((2-nitrophenyl)sulfonyl)oxy)-7-azaspiro[3.5]nonane-7-carboxylate
  • Step 4 tert-butyl 2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2 Synthesis of -carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)-7-azaspiro[3.5]nonane-7-carboxylate
  • Step 5 (2S,4R)-N-(2-((7-azaspiro[3.5]nonan-2-yl)oxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S Synthesis of )-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
  • Step 6 tert-butyl ((1-(5-((2-chloro-3-(3-(2-(2-(((2S,4R)-1-((S)-2-(1- fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)-7-azaspiro[3.5]nonan-7-yl Synthesis of )-3-oxopropanamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate
  • Step 7 (2S,4R)-N-(2-((7-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl) thio)-2-chlorophenyl)amino)-3-oxopropanoyl)-7-azaspiro[3.5]nonan-2-yl)oxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S) Synthesis of -2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
  • Step 1 ethyl 3-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl) Synthesis of amino)-3-oxopropanoate
  • tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (intermediate 3, 240 mg, 0.52 mmol) was suspended in DCM (3.2 ml), and ethyl malonyl chloride (0.08 ml, 0.62 mmol) was slowly added at 0 °C, followed by stirring for 30 minutes. A saturated aqueous solution of NaHCO 3 (10 ml) was added to the reaction solution, followed by extraction with DCM (10 ml x 2).
  • Step 2 3-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino )-3-oxopropanoic acid synthesis
  • Step 3 tert-butyl 4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4- Synthesis of hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-1-carboxylate
  • (2S,4R)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)- 3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (intermediate 10, 100 mg, 0.16 mmol), tert-butyl piperazine-1-carboxylate (38 mg, 0.2 mmol), potassium carbonate (70 mg, 0.5 mmol) , and sodium iodide (26 mg, 0.16 mmol) were suspended in acetonitrile (1 ml) and stirred at 60 °C for 16 hours.
  • Step 4 (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5 Synthesis of -yl)-2-(2-(piperazin-1-yl)ethoxy)benzyl)pyrrolidine-2-carboxamide
  • Step 5 tert-butyl ((1-(5-((2-chloro-3-(3-(4-(2-(2-(((2S,4R)-1-((S)-2- (1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazin-1-yl)- Synthesis of 3-oxopropanamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate
  • Step 6 (2S,4R)-N-(2-(2-(4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2- yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperazin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1 Synthesis of -fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
  • Example 58 was synthesized in the same manner as in Example 52 using ethyl 3-chloro-2-methyl-3-oxopropanoate instead of ethyl malonyl chloride.
  • Example 59 was synthesized in the same manner as in Example 52 using methyl 3-chloro-2,2-dimethyl-3-oxopropanoate instead of ethyl malonyl chloride.
  • Example 62 was synthesized in the same manner as in Example 52, using tert-butyl (R) -2-methylpiperazine-1-carboxylate instead of tert-butyl piperazine-1-carboxylate.
  • Example 63 was synthesized in the same manner as Example 52, using tert-butyl (S) -2-methylpiperazine-1-carboxylate instead of tert-butyl piperazine-1-carboxylate.
  • Example 65 ((2S,4R)-N-(2-(3-((R)-4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin- 2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)-3-methylpiperazin-1-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S) -2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
  • Example 66 (S)-methyl 4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3 -oxopropanoyl)-1-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine- 2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-2-carboxylate
  • Example 66 was synthesized in the same manner as in Example 52, using (S) -1-tert-butyl 3-methyl piperazine-1,3-dicarboxylate instead of tert-butyl piperazine-1-carboxylate.
  • Example 67 methyl (R)-4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl) amino)-3-oxopropanoyl)-1-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)- 4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-2-carboxylate
  • Example 67 was synthesized in the same manner as Example 52, using (R)-1-(tert-butyl) 3-methyl piperazine-1,3-dicarboxylate instead of tert-butyl piperazine-1-carboxylate.
  • Example 68 was synthesized in the same manner as in Example 52, using tert-butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate instead of tert-butyl piperazine-1-carboxylate.
  • Step 1 1-(tert-butyl) 3-methyl (S)-4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido Synthesis of )-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-1,3-dicarboxylate
  • Step 2 methyl (S)-1-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)- Synthesis of 4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-2-carboxylate
  • Step 3 (S)-1-(2-(2-((((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4 Synthesis of -hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-2-carboxylic acid
  • Step 4 (S)-4-(3-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio )-2-chlorophenyl)amino)-3-oxopropanoyl)-1-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3 Synthesis of ,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-2-carboxylic acid
  • Step 5 (S)-4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino) -3-oxopropanoyl)-1-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4- Synthesis of hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-2-carboxylic acid
  • Example 70 (R)-4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino )-3-oxopropanoyl)-1-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4 -hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-2-carboxylic acid
  • Example 69 using 1-(tert-butyl) 3-methyl (R)-piperazine-1,3-dicarboxylate instead of 1-(tert-butyl) 3-methyl (S)-piperazine-1,3-dicarboxylate
  • Example 70 was synthesized in the same manner as in the synthesis method.
  • Step 1 Synthesis of tert-butyl ((1-(5-((3-(2-bromoacetamido)-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate
  • Step 2 tert-butyl ((1-(5-((2-chloro-3-(2-(4-(3-(2-(((2S,4R)-1-((S)-2- (1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)propyl)piperidin-1-yl)acetamido Synthesis of )phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate
  • Step 3 (2S,4R)-N-(2-(3-(1-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2- yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)piperidin-4-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1 Synthesis of -fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
  • Step 1 Synthesis of tert-butyl ((1-(5-((3-acrylamido-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate
  • Step 2 tert-butyl ((1-(5-((2-chloro-3-(3-(4-(2-(2-(((2S,4R)-1-((S)-2- (1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperidin-1-yl)propanamido Synthesis of )phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate
  • Step 3 (2S,4R)-N-(2-(2-(1-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2- yl)thio)-2-chlorophenyl)amino)-3-oxopropyl)piperidin-4-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1 Synthesis of -fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
  • Step 1 Synthesis of tert-butyl 4-(2-(tert-butoxy)-2-oxoethyl)-3-oxopiperazine-1-carboxylate
  • Step 3 Synthesis of 2-(4-(((9H-fluoren-9-yl)methoxy)carbonyl)-2-oxopiperazin-1-yl)acetic acid
  • Step 4 (9H-fluoren-9-yl)methyl 4-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2- Synthesis of yl)thio)-2-chlorophenyl)carbamoyl)piperidine-1-carboxylate
  • Step 5 tert-butyl ((1-(5-((2-chloro-3-(2-(2-oxopiperazin-1-yl)acetamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin- Synthesis of 4-yl)methyl)carbamate
  • Step 6 tert-butyl ((1-(5-((2-chloro-3-(2-(4-(2-(2-(((2S,4R)-1-((S)-2- (1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)-2-oxopiperazin-1- Synthesis of yl)acetamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate
  • Step 7 (2S,4R)-N-(2-(2-(4-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2- yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)-3-oxopiperazin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2 Synthesis of -(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
  • Example 81 N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-5-(4-(2- (2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5- (4-methylthiazol-5-yl)phenoxy)ethyl)piperazin-1-yl)picolinamide
  • Methyl 5-fluoropicolinate 500 mg, 3.22 mmol was suspended in THF (5 ml), and 2N NaOH aqueous solution (4.8 ml, 9.67 mmol) was added thereto, followed by stirring at room temperature for 1 hour.
  • the organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 320 mg (70%) of a white solid.
  • Step 2 Synthesis of tert-butyl ((1-(5-((2-chloro-3-(5-fluoropicolinamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate
  • Step 3 tert-butyl ((1-(5-((2-chloro-3-(5-(4-(2-(2-(((2S,4R)-1-((S)-2- (1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazin-1-yl)picolinamido Synthesis of )phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate
  • Step 4 N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-5-(4-(2-( 2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-( Synthesis of 4-methylthiazol-5-yl)phenoxy)ethyl)piperazin-1-yl)picolinamide
  • Example 82 N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-6-(4-(2- (2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5- (4-methylthiazol-5-yl)phenoxy)ethyl)piperazin-1-yl)nicotinamide
  • Example 82 was synthesized in the same manner as in Example 81 using methyl 6-fluoropicolinate instead of methyl 5-fluoropicolinate.
  • Example 84 was synthesized in the same manner as in Example 81 using 5-chloropyrimidine-2-carboxylic acid instead of 5-fluoropicolinic acid.
  • Example 84 N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-2-(4-(2- (2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5- (4-methylthiazol-5-yl)phenoxy)ethyl)piperazin-1-yl)pyrimidine-5-carboxamide
  • Example 84 was synthesized in the same manner as in Example 81 using 2-chloropyrimidine-5-carboxylic acid instead of 5-fluoropicolinic acid.
  • Example 85 was synthesized in the same manner as in Example 81 using methyl 6-chloropyridazine-3-carboxylate instead of methyl 5-fluoropicolinate.
  • Example 86 N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-2-(4-(2- (2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5- (4-methylthiazol-5-yl)phenoxy)ethyl)piperazin-1-yl)pyrimidine-4-carboxamide
  • Example 86 was synthesized in the same manner as in Example 81 using 2-chloropyrimidine-4-carboxylic acid instead of 5-fluoropicolinic acid.
  • Example 87 N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-5-(2-(2-(((2S ,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5- yl)phenoxy)-7-azaspiro[3.5]nonan-7-yl)pyrimidine-2-carboxamide
  • Example 88 N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-6-(2-(2- (((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4- methylthiazol-5-yl)phenoxy)-7-azaspiro[3.5]nonan-7-yl)pyridazine-3-carboxamide
  • Step 1 tert-butyl (2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine- Synthesis of 2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)carbamate
  • Step 3 methyl 2-(4-(4-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio Synthesis of )-2-chlorophenyl)amino)-4-oxobutyl)piperidin-1-yl)pyrimidine-5-carboxylate
  • Step 4 2-(4-(4-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio) Synthesis of -2-chlorophenyl)amino)-4-oxobutyl)piperidin-1-yl)pyrimidine-5-carboxylic acid
  • Step 5 tert-butyl ((1-(5-((2-chloro-3-(4-(1-(5-((2-(2-(((2S,4R)-1-((S )-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)carbamoyl)pyrimidin Synthesis of -2-yl)piperidin-4-yl)butanamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate
  • Step 6 2-(4-(4-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)- 4-oxobutyl)piperidin-1-yl)-N-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3- Synthesis of dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)pyrimidine-5-carboxamide
  • Step 1 tert-butyl ((1-(5-((2-chloro-3-(4-(1-(5-hydroxypyrimidin-2-yl)piperidin-4-yl)butanamido)phenyl)thio)pyrazin- Synthesis of 2-yl)-4-methylpiperidin-4-yl)methyl)carbamate
  • Step 2 tert-butyl ((1-(5-((2-chloro-3-(4-(1-(5-(2-(2-(((2S,4R)-1-((S) -2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethoxy)pyrimidin-2- Synthesis of yl)piperidin-4-yl)butanamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate
  • Step 3 (2S,4R)-N-(2-(2-((2-(4-(4-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl) pyrazin-2-yl)thio)-2-chlorophenyl)amino)-4-oxobutyl)piperidin-1-yl)pyrimidin-5-yl)oxy)ethoxy)-4-(4-methylthiazol-5-yl)benzyl) -1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
  • KYSE-520 cells were treated with the synthesized example compounds, and the amount of SHP2 protein present in the cells was measured using western blotting detection.
  • the protocol of the experiment using KYSE-520 cells is as follows.
  • KYSE-520 cells were resuspended in RPMI1640 MEDIUM (Hyclone, SH30027.01) containing 5% FBS (Hyclone, SV30207.02) and 1% Penicillin-streptomycin (Welgene, LS 202-02) to form 2.5 x 10 cells. After seeding 2 mL each in a 6-well plate at 5 /well, incubate for 2 days.
  • Iysis buffer is prepared as follows. Add 60 mL of RIPA buffer (Biosesang, RC2038-050-00) + 0.5 mM PMSF (SIGMA, P7626) + 1x Protease/Phosphatase Inhibitor (Cell signaling, 5872S) per well and place on ice for 30 minutes (at 0 and 30 minutes) respectively vortexing). After that, proceed with sonication (10 sec pulse, 30 sec rest, 5 cycles, 70% amplification), centrifuge (15000 g, 15 min, 4°C), and collect only the supernatant and transfer to a new microtube.
  • RIPA buffer Biosesang, RC2038-050-00
  • PMSF SIGMA, P7626
  • 1x Protease/Phosphatase Inhibitor Cell signaling, 5872S
  • Anti SHPTP2 Mouse (1:500 in 5% Skim milk/TBST, size: 70 kDa, Santacruz sc-7384) was incubated overnight at 4°C, and Anti GAPDH Rabbit (1:2,000 in 5% BSA/0.2% TBST, size : 37 kDa, CST #2118) is incubated at room temperature for 1 hour 30 minutes or 3 hours. Wash 3 times for 5 minutes each with 0.2% TBST washing buffer.
  • MV-4-11 cells were treated with the synthesized example compounds, and the amount of SHP2 protein present in the cells was measured using western blotting detection.
  • the protocol of the experiment using MV-4-11 cells is as follows.
  • MV-4-11 cells were resuspended in IMDM MEDIUM (Hyclone, SH30228.01, 5% FBS (Hyclone, SV30207.02), 1% Penicillin-streptomycin (Welgene, LS 202-02)) media and 1x Seed 1 mL each in a 12-well plate at 10 6 /mL.
  • Isis buffer is prepared as follows. Add 50 mL of RIPA buffer (Biosesang, RC2038-050-00) + 0.5 mM PMSF (SIGMA, P7626) + 1x Protease/Phosphatase Inhibitor (Cell signaling, 5872S) per well and leave it on ice for 30 minutes (0, 30 minutes) on each vortexing). After that, proceed with sonication (10 sec pulse, 30 sec rest, 5 cycles, 70% amplification), centrifuge (15000 g, 15 min, 4°C), and collect only the supernatant and transfer to a microtube.
  • RIPA buffer Biosesang, RC2038-050-00
  • PMSF SIGMA, P7626
  • 1x Protease/Phosphatase Inhibitor Cell signaling, 5872S
  • Anti-SHPTP2 Mouse (1:500 in 2% BSA/0.2% TBST, size: 70 kDa, Santacruz sc-7384) was incubated overnight at 4°C, and Anti-GAPDH Rabbit (1:2,000 in 2% BSA/0.2% TBST, size: 37 kDa, CST #2118) is incubated at room temperature for 1 hour 30 minutes or 3 hours.
  • SHP2 protein degrading ability evaluation results of the compounds of the present invention are summarized in Table 4 below.
  • compounds 1-7, 13-19, and 21-26 are the results in KYSE-520 cells, and the others are results in MV-4-11 cells.
  • the compounds according to the present invention showed excellent activity in degrading SHP2 protein.
  • compounds 20, 29-45, 50, 52-58, 60-64, 66-68, 76-80, and 83-85 of the present invention exhibit superior SHP2 proteolytic activity than other compounds having similar structures.
  • the compounds 29, 31, 33, 36-38, 43, 50, 52-58, 61-63, 66, 68, 76-80, and 83 of the present invention showed significantly better SHP2 proteolytic activity than other compounds. showed up

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Abstract

본 개시는 SHP2를 분해하는 측면에서 우수한 활성을 갖는 특정 화학구조의 화합물 또는 이의 약학적으로 허용 가능한 염을 제공한다. 본 개시는 또한 이러한 화합물 또는 이의 약학적으로 허용 가능한 염을 포함하는 조성물을 제공한다. 본 개시는 또한 본 개시에 따른 화합물, 이의 염 및 이들을 포함하는 조성물의 SHP2 관련 질환(예를 들어, 암)의 치료 또는 예방용 의약 용도를 제공한다. 본 개시는 또한 본 개시에 따른 화합물, 이의 염 또는 이들을 포함하는 조성물의 유효한 양을 치료가 필요한 개체에게 투여하는 것을 포함하는 SHP2 관련 질환(예를 들어, 암)의 치료 또는 예방 방법을 제공한다.

Description

SHP2 단백질 분해활성을 갖는 화합물 및 이들의 의약 용도
본 개시는 SHP2 단백질 분해 활성을 가지는 일 군의 화합물에 관한 것이다. 특히, 본 개시는 특정 구조를 가져 SHP2 단백질을 분해하는 활성이 매우 우수한 일 군의 화합물에 관한 것이다. 본 개시는 또한 이러한 화합물을 이용하여, SHP2 단백질 관련 질환을 치료하는 유용한 방법에 관한 것이다. 즉, 본 개시는 본 개시에 따른 화합물들의 SHP2 단백질 관련 질환을 치료 또는 예방하기 위한 의약 용도에 관한 것이다.
Src 상동성 2 도메인-함유 포스파타제(Src homology 2 domain-containing phosphatase, SHP2)는 단백질 티로신 포스파타제이다. SHP2의 돌연변이는 누난(Noonan) 신드롬 및 LEOPARD 신드롬에 널리 퍼져있다. 또 SHP2의 활성화된 돌연변이는 소아 골수단구성 백혈병, 골수이형성 증후군, B-세포 급성 림프모구성 백혈병 및 급성 골수성 백혈병에서도 확인되었다. SHP2의 체세포 활성화 돌연변이는 폐 선암종, 결장암, 신경모세포종, 교모세포종, 흑색종, 간세포 암종, 전립선암 및 유방암을 비롯한 여러 유형의 고형 종양과 관련이 있다 (Bentires-Alj et al., Cancer. Res. 2004, 64, 8816-8820).
SHP2가 암 세포 내에서 RAS-ERK, JAK-STAT, PI3K-AKT, NF-κB 및 mTOR 경로와 같은 다양한 신호 전달 프로세스에 관여한다는 것은 입증되었다. RAS-ERK 경로에서 SHP2는 RAS-RAF-MEK-ERK 키나제 캐스케이드 신호 전달을 촉진하기 위해 업스트림에서 양성 조절자 역할을 한다. 따라서 SHP2 억제는 ERK의 탈인산화 및 RAS-RAF-MEK-ERK 경로의 발암성 기능을 저해하여, 결과적으로 암세포에서 세포 성장 억제 및 세포자멸사를 유도한다. SHP2는 또한 프로그램된 세포 사멸 경로(PD-1/PD-L1)에 참여하고 T 세포 활성화를 억제하여 면역 회피에 기여한다. 요약하면, SHP2는 매우 매력적인 암 치료 표적이다. 예를 들어, SHP2 억제제인 SHP099는 SHP2 포스파타제 활성을 선택적으로 차단하고 시험관 내 암세포 성장과 이종이식 마우스 모델에서 종양 성장을 억제하는 것으로 나타났다 (Chen et al., Nature 2016, 535, 148-152; Garcia Fortanet et al., J. Med. Chem. 2016, 59, 7773-7782).
알로스테릭 SHP2 억제제는 Kirsten 래트 육종(KRAS)-돌연변이 인간 암의 전임상 모델에서 효과적인 것으로 나타났다. SHP2 단백질의 제거는 또한 SHP2 활성 억제를 위한 대안적이고 아마도 훨씬 더 효과적인 전략을 제공할 수 있다. 예를 들어, CRISPR-Cas9에 의한 SHP2 비활성화는 KRAS-돌연변이 종양의 이종이식 모델에서 노화를 유도하고 종양의 성장을 억제한다는 것이 입증되었다. 이러한 발견은 종양 세포에서 SHP2 단백질의 제거가 인간 암, 특히 KRAS-돌연변이를 갖는 암에 대한 효과적인 치료 전략이 될 수 있다는 증거를 제공한다.
따라서 본 발명이 해결하고자 하는 과제는 Src 상동성 2 도메인-함유 포스파타제(Src homology 2 domain-containing phosphatase, SHP2) 분해 활성을 가지는 화합물, 이들을 유효성분으로 포함하는 약학 조성물, 및 이들의 SHP2 관련 질환 (바람직하게는, 암 또는 종양) 치료 또는 예방용 의약 용도를 제공하는 것이다.
본 발명이 해결하고자 하는 다른 과제는 SHP2를 분해하여 결과적으로 SHP2 활성을 낮추는 것을 특징으로 하는, 본 발명에 따른 화합물을 SHP2 관련 질환의 치료, 개선 또는 예방이 필요한 환자에게 투여하는 것을 특징으로 하는 SHP2 관련 질환(바람직하게는, 암 또는 종양)의 치료 또는 개선 방법을 제공하는 것이다.
본 발명 화합물
상기 해결하고자 하는 과제를 달성하기 위하여, 일 양태에서, 본 발명은 하기 화학식 1의 화합물 또는 이의 약학적으로 허용 가능한 염을 제공한다.
Figure PCTKR2022011797-appb-img-000001
상기 화학식 1에서,
R1a, R1b, R1c, 및 R1d는 서로 독립적으로 H 또는 C1-6alkyl (바람직하게는 메틸)이고,
R2는 H, C1-6alkyl, 또는 -NH2이고,
R3는 H, C1-6alkyl, 또는 C1-6hydroxyalkyl이고,
R4는 H, C1-6alkyl, halogen (바람직하게는 Cl), 또는 C1-6haloalkyl이고,
R5는 H, C1-6alkyl, C1-6alkynyl, halogen, -CN, 또는 heteroaryl (바람직하게는 thiazole)이며, 여기에서 헤테로아릴은 임의로 고리 내 하나 이상의 수소가 C1-6alkyl (바람직하게는 메틸), halogen, 또는 C1-3haloalkyl로 치환되며,
R6는 -NHC(O)R7 또는 heteroaryl이고, 여기에서 헤테로아릴은 임의로 고리 내 하나 이상의 수소가 C1-6alkyl, halogen, 또는 C1-3haloalkyl로 치환되며,
R7는 C1-6alkyl (바람직하게는 메틸), C1-6haloalkyl, C3-10사이클로알킬 (바람직하게는 사이클로프로필), 또는 헤테로사이클이고, 여기에서 사이클로알킬 및 헤테로사이클은 서로 독립적이면서 임의로 고리 내 하나 이상의 수소가 C1-6alkyl, halogen (바람직하게는 F), 또는 -CN로 치환되며,
X는 CH 또는 N이고,
n1 및 n2는 서로 독립적으로 0, 1, 또는 2이고,
m은 0 또는 1이며,
L은 하기 화학식 2이며,
Figure PCTKR2022011797-appb-img-000002
화학식 2에서,
A1, A2, 및 A3은 서로 독립적으로 직접 결합, -O-, -N(R9)- -C(O)-, -C(O)N(R9)-, 또는 -N(R9)C(O)-이고,
B1 및 B2는 서로 독립적으로 직접 결합, C3-10사이클로알킬, 헤테로사이클 (바람직하게는 아제티딘, 피롤리딘, 피페리딘, 피페라진, azaspiro[3.5]nonane, diazabicyclo[3.2.1]octane), 아릴, 또는 헤테로아릴 (바람직하게는 피리딘, 피리미딘, 피리다진, 피라진)이며, 여기에서 사이클로알킬, 헤테로사이클, 아릴, 또는 헤테로아릴은 임의로 고리 내 하나 이상의 수소가 C1-6alkyl (바람직하게는 메틸), halogen, C1-6haloalkyl, -OH, =O, 또는 -CO2R9로 치환되며,
R8 및 R8'는 서로 독립적으로 H 또는 C1-6alkyl이거나, R8 및 R8'는 서로 연결되어 C3-10사이클로알킬 (바람직하게는 사이클로프로필)을 형성하고,
R9는 수소 또는 C1-6alkyl (바람직하게는 메틸)이고,
q1, q2, q3, q4, 및 q5는 서로 독립적으로 0 내지 10의 정수임.
본 발명에 따른 화합물은 'SHP2 결합 리간드-링커-E3 리가아제 리간드'(화학식 1에서 각각 'L 기준 왼쪽 모이어티-링커(L)-L 기준 오른쪽 모이어티')의 프로탁(PROTAC, Proteolysis-targeting chimera) 화합물이다. 본 발명자들은 본 발명의 화학식 1의 SHP2 결합 모이어티에 CRBN 리간드, VHL 리간드, IAP 리간드 등 다양한 E3 리가아제 리간드를 도입해 보았으며, 이들 리간드 중에서도 특이하게 본 발명의 화학식 1과 같은 VHL 리간드만 그 효과가 우수하였다. 또한 VHL 리간드를 사용하는 경우에 있어서도 링커(화학식 1의 L)가 VHL 리간드 내 페닐 기에 연결되는 경우에 특이하게도 그 효과가 탁월하였다. 본 발명은 이러한 특이하고도 놀라운 발견에 기초하나, 본 발명은 이러한 이론적 기전에 한정되는 것은 아니다.
본 발명자들은 다양한 치환기들을 평가하였으며, 본 개시의 화학식 1과 같은 구조일 경우, 특히, 후술하는 바람직한 구조일 경우 SHP2 분해 활성을 비롯한 여러 측면에서 우수한 화합물을 얻을 수 있었다. 즉, 본 발명은 SHP2 분해 활성, (대사)안정성 등이 우수하고, 약효 성분으로서의 물리화학적 성질(cLogP value, 수용해성, 세포막 투과성) 등이 우수한 새로운 화합물을 제공한다.
본 명세서에서 용어 "치환기(substituent)", "라디칼(radical)", "기(group)", "모이어티(moiety)", 및 "절편(fragment)"은 서로 바꾸어 사용할 수 있다.
만약 치환기가 "임의로 치환된" 또는 “선택적으로 치환된”으로 설명된다면, 상기 치환기는 (1) 치환되지 않거나 (2) 또는 정의된 치환기들 중 하나 이상으로 치환될 수 있다. 만약 치환 가능한 위치가 치환되지 않은 경우 기본(default) 치환기는 수소이다.
본 명세서에서 사용된 용어 "알킬"은 (탄소수가 특별히 한정되지 않은 경우) 탄소수 1 내지 10을 가진 포화된 직쇄상 또는 분지상의 비-고리(cyclic) 탄화수소를 의미한다. "저급 알킬"은 탄소수가 1 내지 4인 직쇄상 또는 분지상 알킬을 의미한다. 대표적인 포화 직쇄상 알킬은 -메틸, -에틸, -n-프로필, -n-부틸, -n-펜틸, -n-헥실, -n-헵틸, -n-옥틸, -n-노닐 과 -n-데실을 포함하고, 반면에 포화 분지상 알킬은 -이소프로필, -sec-부틸, -이소부틸, -tert-부틸, 이소펜틸, 2-메틸헥실, 3-메틸부틸, 2-메틸펜틸, 3-메틸펜틸, 4-메틸펜틸, 2-메틸헥실, 3-메틸헥실, 2-메틸펜틸, 3-메틸펜틸, 4-메틸펜틸, 2-메틸헥실, 3-메틸헥실, 4-메틸헥실, 5- 메틸헥실, 2,3-디메틸부틸, 2,3-디메틸펜틸, 2,4-디메틸펜틸, 2,3-디메틸헥실, 2,4-디메틸헥실, 2,5-디메틸헥실, 2,2-디메틸펜틸, 2,2-디메틸헥실, 3,3-디메틸펜틸, 3,3-디메틸헥실, 4,4-디메틸헥실, 2-에틸펜틸, 3-에틸펜틸, 2-데틸헥실, 3-에틸헥실, 4-에틸헥실, 2-메틸-2-에틸펜틸, 2-메틸-3-에틸펜틸, 2-메틸-4-에틸펜틸, 2-메틸-2-에틸헥실, 2-메틸-3-에틸헥실, 2-메틸-4-에틸헥실, 2,2-디에틸펜틸, 3,3-디에틸헥실, 2,2-디에틸헥실, 및 3,3-디에틸헥실을 포함한다. 본 발명의 바람직한 일 양태에서, 알킬은 메틸이다.
본 명세서에서 사용된 용어 "알키닐"은 (탄소수가 특별히 한정되지 않은 경우) 2 내지 10개의 탄소원자를 가지고 있고, 적어도 하나의 탄소-탄소 삼중결합을 포함한 직쇄상 또는 곁가지의 비-고리 탄화수소를 의미한다. 대표적인 직쇄상 또는 분지상 (C2-C10)알키닐은 -아세티레닐, -프로피닐, -1-부티닐, -2-부티닐, -1-펜티닐, -2-펜티닐, -3-메틸-1-부티닐, -4-펜티닐, -1-헥시닐, -2-헥시닐, -5-헥시닐, -1-헵티닐, -2-헵티닐, -6-헵티닐, -1-옥티닐, -2-옥티닐, -7-옥티닐, -1-노니닐, -2-노니닐, -8-노니닐, -1-데시닐, -2-데시닐, 및 -9-데시닐을 포함한다. 이러한 알키닐 그룹은 선택적으로 치환될 수 있다.
본 명세서에서 “C1-6”, "C1-6", 또는 "C1-C6"와 같이 기재될 경우 이는 탄소수가 1 내지 6개임을 의미한다. 예를 들어, C1-6알킬은 탄소수가 1 내지 6인 알킬을 의미한다.
본 명세서에서 사용된 용어 "할로겐" 및 "할로"는 플루오린, 클로린, 브로민 또는 아이오딘을 의미한다. 본 발명의 바람직한 일 양태에 있어, 할로겐은 클로린 또는 플루오린이다.
본 명세서에서 사용된 용어 "할로알킬"은 각각 하나 이상의 수소 원자가 할로겐 원자로 치환된 알킬 그룹을 의미한다. 예를 들어, 할로알킬은 -CF3, -CHF2, -CH2F, -CBr3, -CHBr2, -CH2Br, -CC13, -CHC12, -CH2CI, -CI3, -CHI2, -CH2I, -CH2-CF3, -CH2-CHF2, -CH2-CH2F, -CH2-CBr3, -CH2-CHBr2, -CH2-CH2Br, -CH2-CC13, -CH2-CHC12, -CH2-CH2CI, -CH2-CI3, -CH2-CHI2, -CH2-CH2I, 및 이와 유사한 것을 포함한다. 본 발명의 바람직한 일 양태에서, 할로알킬은 CF3이다. 여기에서 알킬 및 할로겐은 위에서 정의된 것과 같다.
본 명세서에서 사용한 용어 "하이드록시알킬"은 하나 이상의 하이드록시 그룹으로 치환된 선형 또는 분지형 C1-10알킬을 나타내며, 하이드록시알킬 그룹의 예에는 하이드록시메틸, 하이드록시에틸, 하이드록시프로필 및 하이드록시부틸이 포함되지만, 이에 한정되는 것은 아니다.
본 명세서에서 사용된 용어 "사이클로알킬(cycloalkyl)"은 탄소 및 수소 원자를 가지며 탄소-탄소 다중 결합을 가지지 않는 모노사이클릭 또는 폴리사이클릭 포화 고리(ring)를 의미한다. 모노사이클릭 고리의 예는 (C3-C7)사이클로알킬 (예를 들어, 사이클로프로필, 사이클로부틸, 사이클로펜틸, 사이클로헥실 및 사이클로헵틸)을 포함하나 이에 한정되는 것은 아니다. 폴리사이클릭 고리의 예는 octahydropentalene, decahydronaphthalene 등과 같은 융합된(fused) 바이사이클릭(bicyclic) 고리; spiro[3.3]heptane, spiro[3.4]octane, spiro[3.5]nonane, spiro[4.4]nonane, spiro[4.5]decane, spiro[5.5]undecane 등과 같은 스피로 고리; 및 bicyclo[2.1.1]hexane, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane 등과 같은 가교된(bridged) 바이사이클릭 고리를 포함하나 이에 한정되는 것은 아니다. 사이클로알킬 그룹은 선택적으로 치환될 수 있다. 일 실시예에서, 사이클로알킬 그룹은 모노사이클릭 링(고리)이다. 본 발명의 바람직한 일 양태에서, 사이클로알킬은 사이클로프로필이다.
본 명세서에서 사용된 "헤테로사이클(헤테로고리)" 또는 “헤테로사이클로알킬”은 질소, 산소 및 황으로부터 독립적으로 선택된 1 내지 4의 헤테로원자를 함유하는 포화된 5- 내지 7-멤버의 모노사이클릭, 또는 7- 내지 12-멤버의 바이사이클릭 링(고리)를 의미하며, 여기에서 질소 및 황 헤테로원자는 선택적으로 산화될 수 있고, 질소 헤테로원자는 선택적으로 사가화(quaternized)될 수 있다. 대표적인 헤테로고리는 옥시란(oxiran), 옥세탄(oxetan), 테트라하이드로퓨란(tetrahydrofuran), 테트라하이드로피란(tetrahydropyran), 1,4-디옥산(1,4-dioxane), 아지리딘(aziridine), 아제티딘(azetidine), 피롤리딘(pyrrolidine), 피페리딘(piperidine), 피페라진(piperazine), 피롤리디논(pyrrolidinone), 히단토인(hydantoine), 발레롤락탐(valerolactam), 티이란(thiirane), 티에탄(thietane), 테트라하이드로티오펜(tetrahydrothiophene), 테트라하이드로티오피란(tetrahydrothiopyra), 모포린(morpholine), 테트라하이드로피리딘(tetrahydropyridine), 테트라하이드로피리미딘(tetrahydropyrimidine) 등을 포함한다. 헤테로사이클에는 헤테로고리 중 일부가 벤젠 또는 cyclopenta-1,3-diene 고리에 융합한 바이사이클릭 링이 포함된다. 헤테로고리는 헤테로원자 또는 탄소 원자에 의하여 부착될 수 있다. 또한, 헤테로사이클에는 앞서 언급된 폴리사이클릭 고리의 1개 이상의 탄소 원자가 질소, 산소 또는 황 원자로 치한된 융합된(fused) 바이사이클릭(bicyclic) 고리, 스피로 고리 및 가교된(bridged) 바이사이클릭 고리가 포함된다. 이러한 예로는, 예를 들어, 헤테로원자가 질소일 경우 octahydrocyclopenta[c]pyrrole, octahydropyrrolo[3,4-c]pyrrole, decahydroisoquinoline, decahydro-2,6-naphthyridine 등과 같은 융합된(fused) 헤테로바이사이클릭(bicyclic) 고리; 2-azaspiro[3.3]heptane, 2,6-diazaspiro[3.3]heptane, 2-azaspiro[3.4]octane, 2,6-diazaspiro[3.4]octane, 2-azaspiro[3.5]nonane, 2,7-diazaspiro[3.5]nonane, 2-azaspiro[4.4]nonane, 2,7-diazaspiro[4.4]nonane, 8-azaspiro[4.5]decane, 2,8-diazaspiro[4.5]decane, 3-azaspiro[5.5]undecane, 3,9-diazaspiro[5.5]undecane 등과 같은 스피로 고리; 및 2-azabicyclo[2.1.1]hexane, 2-azabicyclo[2.2.1]heptane, 2,5-diazabicyclo[2.2.1]heptane, 2-azabicyclo[2.2.2]octane, 2,5-diazabicyclo[2.2.2]octane 등과 같은 가교된(bridged) 헤테로바이사이클릭 고리를 포함하나 이에 한정되는 것은 아니다. 본 발명의 바람직한 일 양태에서, 헤테로사이클은 아제티딘, 디아제티딘, 피페리딘, 피페라진, azaspiro[3.5]nonane, diazabicyclo[3.2.1]octane 등에서 선택된 어느 하나이다.
본 명세서에서 사용된 용어 "아릴"은 5 내지 10의 고리 원자를 함유하는 탄소고리 방향족 그룹을 의미한다. 대표적인 예는 페닐, 톨일(tolyl), 자이릴(xylyl), 나프틸, 테트라하이드로나프틸, 안트라세닐(anthracenyl), 플루오레닐(fluorenyl), 인데닐(indenyl), 아주레닐(azulenyl) 등을 포함하나 이에 한정되는 것은 아니다. 탄소고리 방향족 그룹은 선택적으로 치환될 수 있다.
본 명세서에서 사용된 "헤테로아릴"은 질소, 산소 및 황으로 구성된 군으로부터 선택된 적어도 하나의 헤테로원자를 가지고, 모노- 및 바이사이클릭 링 시스템을 포함하는 적어도 하나의 탄소 원자를 포함하는 5 내지 10 멤버의 방향족 헤테로고리(heterocycle) 링이다. 대표적인 헤테로아릴은 furan, 4H-pyran, pyrrole, imidazole, pyrazole, triazole, tetrazole, pyridine, pyrimidine, pyridazine, pyrazine, triazine, thiophene, ozaxole, isoxazole, thiazole, isothiazole, oxadiazole, benzofuran, benzothiophene, quinoline, indole, benzoxazole, benzimidazole, benzothiazole, cinnoline, phthalazine, quinazoline, 1H-azepine 등이다. 본 발명의 바람직한 일 양태에서, 헤테로아릴은 thiazole, 피리딘, 또는 피라진이다.
본 명세서에 있어, * 또는
Figure PCTKR2022011797-appb-img-000003
은 다른 moiety와 연결되어 있는 것을 의미한다.
바람직하게, SHP2 분해 활성, (대사)안정성, 물리화학적 특성 등 여러 측면에서, 본 발명의 바람직한 일 양태는 상기 화학식 1에서,
R1a, R1b, R1c, 및 R1d는 서로 독립적으로 H 또는 C1-6alkyl (바람직하게는 메틸)이고,
R2는 H, C1-6alkyl, 또는 -NH2이고,
R3는 H, C1-6alkyl, 또는 C1-6hydroxyalkyl이고,
R4는 H, C1-6alkyl, 또는 halogen (바람직하게는 Cl)이고,
R5는 heteroaryl (바람직하게는 thiazole)이며, 여기에서 헤테로아릴은 임의로 고리 내 하나 이상의 수소가 C1-6alkyl (바람직하게는 메틸) 또는 halogen으로 치환되며,
R6는 -NHC(O)R7이고,
R7는 C1-6alkyl (바람직하게는 메틸) 또는 C3-10사이클로알킬 (바람직하게는 사이클로프로필)이고, 여기에서 사이클로알킬은 임의로 고리 내 하나 이상의 수소가 C1-6alkyl, halogen (바람직하게는 F), 또는 -CN로 치환되며,
X는 CH 또는 N이고,
n1 및 n2는 서로 독립적으로 0 또는 1이고,
m은 0 또는 1이며,
L은 하기 화학식 2이며,
[화학식 2]
Figure PCTKR2022011797-appb-img-000004
화학식 2에서,
A1, A2, 및 A3은 서로 독립적으로 직접 결합, -O-, -N(R9)- -C(O)-, -C(O)N(R9)-, 또는 -N(R9)C(O)-이고,
B1 및 B2는 서로 독립적으로 직접 결합, 헤테로사이클 (바람직하게는 아제티딘, 피롤리딘, 피페리딘, 피페라진, azaspiro[3.5]nonane, diazabicyclo[3.2.1]octane), 또는 헤테로아릴 (바람직하게는 피리딘, 피리미딘, 피리다진, 피라진)이며, 여기에서 헤테로사이클 또는 헤테로아릴은 임의로 고리 내 하나 이상의 수소가 C1-6alkyl (바람직하게는 메틸), halogen, -OH, =O, 또는 -CO2R9로 치환되며,
R8 및 R8'는 서로 독립적으로 H 또는 C1-6alkyl이거나, R8 및 R8'는 서로 연결되어 C3-10사이클로알킬 (바람직하게는 사이클로프로필)을 형성하고,
R9는 수소 또는 C1-6alkyl (바람직하게는 메틸)이고,
q1, q2, q3, q4, 및 q5는 서로 독립적으로 0 내지 10의 정수인,
화합물 또는 이의 약학적으로 허용 가능한 염을 제공한다.
보다 바람직하게, SHP2 분해 활성, (대사)안정성, 물리화학적 특성 등 여러 측면에서, 특히, SHP2 분해 활성 측면에서 본 발명의 더욱 바람직한 일 양태는 상기 화학식 1에서,
R1a, R1b, R1c, 및 R1d는 서로 독립적으로 H 또는 C1-6alkyl (바람직하게는 메틸)이고,
R2는 H 또는 -NH2이고,
R3는 H이고,
R4는 H 또는 halogen (바람직하게는 Cl)이고,
R5는 thiazole이며, 여기에서 thiazole은 임의로 고리 내 하나 이상의 수소가 C1-6alkyl (바람직하게는 메틸)으로 치환되며,
R6는 -NHC(O)R7이고,
R7는 C1-6alkyl (바람직하게는 메틸) 또는 사이클로프로필이고, 여기에서 사이클로프로필은 임의로 고리 내 하나 이상의 수소가 halogen (바람직하게는 F) 또는 -CN으로 치환되며,
X는 CH이고,
n1 및 n2는 서로 독립적으로 0 또는 1이고,
m은 0 또는 1이며,
L은 하기 화학식 2이며,
[화학식 2]
Figure PCTKR2022011797-appb-img-000005
화학식 2에서,
A1, A2, 및 A3은 서로 독립적으로 직접 결합, -O-, -N(R9)- -C(O)-, -C(O)N(R9)-, 또는 -N(R9)C(O)-이고,
B1 및 B2는 서로 독립적으로 직접 결합, 아제티딘, 피롤리딘, 피페리딘, 피페라진, azaspiro[3.5]nonane, diazabicyclo[3.2.1]octane, 피리딘, 피리미딘, 피리다진, 또는 피라진이며, 여기에서 아제티딘, 피롤리딘, 피페리딘, 피페라진, azaspiro[3.5]nonane, diazabicyclo[3.2.1]octane, 피리딘, 피리미딘, 피리다진, 또는 피라진은 임의로 고리 내 하나 이상의 수소가 C1-6alkyl (바람직하게는 메틸), -OH, =O, 또는 -CO2R9로 치환되며,
B1 및 B2 중 어느 하나 이상은 아제티딘, 피롤리딘, 피페리딘, 피페라진, azaspiro[3.5]nonane, diazabicyclo[3.2.1]octane, 피리딘, 피리미딘, 피리다진, 또는 피라진이고,
R8 및 R8'는 서로 독립적으로 H 또는 C1-6alkyl이거나, R8 및 R8'는 서로 연결되어 C3-10사이클로알킬 (바람직하게는 사이클로프로필)을 형성하고,
R9는 수소 또는 C1-6alkyl (바람직하게는 메틸)이고,
q1, q2, q3, q4, 및 q5는 서로 독립적으로 0 내지 10의 정수인,
화합물 또는 이의 약학적으로 허용 가능한 염을 제공한다.
바람직하게, 본 발명에 따른 SHP2 프로탁 화합물에 있어, 링커(L)은 하기 링커들 중 어느 하나이다. 하기와 같은 링커를 가질 경우 SHP2 분해 활성이 우수하며, 본 발명의 다양한 목적에 더욱 적합하다.
Figure PCTKR2022011797-appb-img-000006
Figure PCTKR2022011797-appb-img-000007
비-한정적인, 본 개시에 따른 화학식 1의 화합물의 예는 후술하는 실시예들에서 제조된 화합물들이다. 각 실시예 번호는 화합물 번호에 대응한다. 예를 들어, 실시예 10에서 제조된 최종 화합물의 번호는 화합물 10이다.
상기 화합물들 중에서도, 특히 하기 표 1의 화합물들이 SHP2 분해 활성, 암 세포주 세포독성, (대사)안정성, 물리화학적 성질 등 다양한 측면에서, 특히, SHP2 분해 활성 측면에서 더욱 바람직하였다.
Figure PCTKR2022011797-appb-img-000008
Figure PCTKR2022011797-appb-img-000009
Figure PCTKR2022011797-appb-img-000010
본 발명에 있어 "약학적으로 허용 가능한 염"은 여기서 언급한 화합물들에서 발견되는 특정 치환체에 의존하는 비교적 비독성 산 및 염기로 제조된 활성 화합물의 염들을 포함한다. 본 발명의 화합물들은 상대적으로 산성 기능성을 포함할 때, 염기(base) 부가 염들은 충분한 양의 원하는 염기, 순수한 또는 적당한 비활성(inert) 용매로 그러한 화합물들의 중성 형태를 접촉하여 얻을 수 있다. 약학적으로 허용 가능한 염기 부가 염의 예들은 나트륨, 칼륨, 칼슘, 암모늄, 유기 아미노 또는 마그네슘 염 또는 유사한 염을 포함한다. 본 발명의 화합물들은 상대적으로 염기성 기능성을 포함할 때, 산성 부가 염들은 충분한 양의 원하는 산, 순수한 또는 적당한 비활성(inert) 용매로 그러한 화합물들의 중성 형태를 접촉하여 얻을 수 있다. 약학적으로 허용 가능한 산성 부가 염의 예들은 초산, 프로피온산, 이소부틸산, 옥살릭산(oxalic), 마레익(maleic), 말로닉(malonic), 안식향성, 숙신산, 수버릭(suberic), 푸마릭(fumaric), 만데릭(mandelic), 프탈릭(phthalic), 벤젠설포닉(benzenesulfonic), p-토릴설포닉(tolylsulfonic), 구연산, 주석산, 메탄솔포닉(methanesulfonic), 및 그 유사체를 포함하는 상대적으로 비독성 유기산에서 유래한 염들 뿐만 아니라, 염화수소, 브롬화 수소, 질산, 탄산, 일수소탄산(monohydrogencarbonic), 인산(phosphoric), 일수소인산, 이수소인산, 황산, 일수소황산, 요오드화수소 또는 아인산(phosphorous acid) 및 그 유사체를 포함한다. 또한 알긴네이트(arginate)와 그 유사체와 같은 아미노산의 염 및 글루쿠로닉(glucuronic) 또는 갈락투노릭(galactunoric) 산들과 그 유사체와 같은 유기산의 유사체를 포함한다. 본 발명의 일부 특정한 화합물들은 화합물들을 염기성 또는 산성 부가(addition) 염들로 전환하게 하는 염기성 및 산성 기능성 모두를 갖는다. 염들의 다른 예들은 본 발명이 속한 분야에서 공지된 문헌들을 통해 잘 알려져 있다.
본 명세서에서 사용된 용어인 "본 발명의 화합물"은 화학식 1 각각의 화합물들뿐만 아니라, 이들의 클라드레이트(clathrates), 수화물, 용매화물, 또는 다형체를 포함하는 의미이다. 또한 용어 “본 발명의 화합물”은 이의 약학적으로 허용 가능한 염이 언급되지 않을 경우 본 발명 화합물의 약학적으로 허용 가능한 염도 포함하는 의미이다. 일 실시예에 본 발명의 화합물은 입체이성질체적으로 순수한 화합물들(예를 들어, 다른 입체이성질체가 실질적으로 없는(예를 들어, 85% ee 이상, 90% ee 이상, 95% ee 이상, 97% ee 이상, 또는 99% ee 이상))로 존재할 수 있다. 즉, 본 발명에 따른 화학식 1의 화합물 또는 그의 염이 호변이성적(tautomeric) 이성질체 및/또는 입체이성질체(예를 들어, 기하이성질체(geometrical isomer) 및 배좌 이성질체(conformational isomers))일 경우 그들의 분리된 이성질체 및 혼합물 각각 또한 본 발명의 화합물의 범주에 포함된다. 본 발명의 화합물 또는 그의 염이 구조 내에 비대칭 탄소(asymmetric carbon)를 가지고 있는 경우에, 그들의 광학 활성 화합물 및 라세믹 혼합물들 또한 본 발명의 화합물의 범위에 포함된다.
본 명세서에서 사용될 경우, 용어 "결정다형(polymorph)"은 본 발명의 화합물의 고체 결정 형태 또는 그것의 복합체를 의미한다. 같은 화합물의 다른 결정다형은 다른 물리적, 화학적 그리고/또는 스펙트럼적 특성을 보인다. 물리적 특성 측면의 차이점으로는 안정성(예를 들어, 열 또는 빛 안정성), 압축성과 밀도(제제화 및 생산물 제조에 중요함), 그리고 용해율(생물학적 이용률에 영향을 줄 수 있음)을 포함하나, 이에 한정되지 아니한다. 안정성에서 차이는 화학반응성 변화들(예를 들어, 또 다른 다형으로 구성되었을 때보다 하나의 다형으로 구성되었을 때 더 빠르게 변색이 되는 것 같은 차별적 산화) 또는 기계적인 특징들(예를 들어 동역학적으로 선호된 다형체로서 저장된 정제 파편들이 열역학 적으로 더 안정된 다형으로 변환) 또는 둘 다(하나의 다형의 정제는 높은 습도에서 더 분해에 예민)를 야기한다. 결정다형의 다른 물리적 성질들은 그들의 가공에 영향을 줄 수 있다. 예를 들어, 한 결정다형은 또 다른 결정다형에 비하여, 예를 들어, 그것의 형태 또는 입자의 크기 분포에 기인하여 용매화합물을 형성할 가능성이 많을 수 있거나, 여과 또는 세척이 더 어려울 수 있다.
본 명세서에서 사용된 용어 "용매 화합물"은 비공유 분자간의 힘에 의해 결합된 화학량론적 또는 비-화학량론적인 양의 용매를 포함하는 본 발명의 화합물 또는 이의 약학적으로 허용 가능한 염을 의미한다. 바람직한 용매들은 휘발성이고, 비독성이며, 인간에게 극소량 투여될 수 있다.
본 명세서에서 사용된 용어 "수화물(hydrate)"은 비공유 분자간의 힘에 의해 결합된 화학량론적 또는 비-화학량론적인 양의 물을 포함하는 본 발명의 화합물 또는 이의 약학적으로 허용 가능한 염을 의미한다.
본 명세서에서 사용된 용어 "클라드레이트(clathrate)"은 게스트 분자(예를 들어, 용매 또는 물)를 가두어 놓은 공간(예를 들어, 채널(channel))을 포함한 결정 격자의 형태의 본 발명의 화합물 또는 그것의 염을 의미한다.
본 명세서에서 사용된 용어 "정제된(purified)"은 분리될 때, 분리체는 90% 이상 순수한 것을 의미하며, 일 실시예에서는 95% 이상 순수하고, 다른 실시 예에서는 99% 이상 순수하고, 또 다른 실시예에서는 99.9% 이상 순수한 것을 의미한다.
본 발명 화합물의 의약 용도 및 치료 방법
본 발명은 하나 이상의 상기와 같은 화합물의 치료적으로 유효한 양을 개체에게 투여함으로써 하기 질병 또는 상태(condition)를 갖거나 갖기 쉬운 개체에서 하기 질병 또는 상태(condition)를 치료하는 방법을 더 제공한다. 일 양태에서, 상기 치료는 예방 치료(preventative treatment)이다. 또 다른 양태에서, 상기 치료는 완화 치료(palliative treatment)이다. 또 다른 양태에서, 상기 치료는 회복 치료(restorative treatment)이다.
1. 질병 또는 상태(Condition)
본 발명의 SHP2 분해용 화합물들은 다양한 치료학적 또는 예방학적 용도(예를 들어, 암)에 유용하다. 이러한 화합물들은 SHP2을 분해하여 SHP2 활성을 낮추기 위해 사용될 수 있으며, 또 SHP2 관련 질환의 치료를 위해서 또는 이러한 질병의 악화를 방지하기 위하여 사용될 수 있다. 따라서 본 발명은 세포 내 SHP2을 분해하는 방법을 제공한다. 이러한 방법에서 상기 세포는 본 발명의 화합물의 유효한 양과 접촉하게 된다. 일 실시예에서, 상기 세포는 개체 내에 존재한다. 본 발명의 방법은 치료 또는 예방이 필요한 개체에게 치료적으로 또는 예방학적으로 유효한 양의 본 발명에 따른 화합물을 포함하는 약학 조성물을 투여하는 것을 포함한다.
일 양태에서, 본 발명은 SHP2 관련 질환의 세포 내에서 SHP2을 분해하는 방법을 제공한다. 예를 들어, 본 발명은 후술하는 SHP2 관련 질환을 가진 개체의 세포 내에서 SHP2을 분해하여 결과적으로 SHP2 활성을 낮추기 위하여 이용될 수 있다. 본 발명의 다른 양태에서, 본 발명은 암, 특히 백혈병, 림프종, 폐암, 두경부암, 식도암, 위암, 대장암, 췌장암, 간암, 유방암, 난소암, 자궁경부암, 방광암, 흑색종, 신경모세포종, 신경교종, 육종 등의 세포 내에서 SHP2를 분해하기 위하여 이용될 수 있다.
다른 양태에서, 본 발명은 화학식 1의 화합물 또는 이의 약학적으로 허용 가능한 염의 치료적으로 유효한 양을 개체에게 투여하는 단계를 포함하는 SHP2 관련 질환을 치료하는 방법을 제공한다. 이러한 방법은 SHP2를 분해하기 위해 충분한 양, 즉, 치료학적으로 유효한 양의 본 발명 화합물을 치료가 필요한 개체에게 투여하는 단계를 포함한다. 이러한 방법에 있어, 본 발명의 화합물은 본 명세서에서 설명되는 약학 조성물의 형태로 상기 개체에 투여될 수 있다.
본 발명에 있어, SHP2 관련 질환은 암이다. 예를 들어, 국제특허출원 공개공보 WO 2021/236775의 표 2에 기재된 암들 중 어느 하나 일 수 있지만 이에 제한되지 않는다. WO 2021/236775의 표 2의 기재는 인용에 의해 본 명세서에 모두 포함된다. 또 다른 양태에서, 암은 고형 종양이다. 또 다른 양태에서, 암은 혈액암이다. 예시적인 혈액암은 WO 2021/236775의 표 3에 열거된 암을 포함하지만 이에 제한되지 않는다. WO 2021/236775의 표 3의 기재는 인용에 의해 본 명세서에 모두 포함된다. 또 다른 양태에서, 혈액암은 급성 림프구성 백혈병, 만성 림프구성 백혈병(B-세포 만성 림프구성 백혈병 포함), 또는 급성 골수성 백혈병이다. 또 다른 양태에서, 암은 백혈병, 예를 들어 급성 단핵구 백혈병, 급성 골수성 백혈병, 만성 골수성 백혈병, 만성 림프구성 백혈병 및 혼합 계통 백혈병(MLL)으로부터 선택된 백혈병이다. 또 다른 양태에서, 암은 NUT-정중선 암종이다. 또 다른 양태에서, 암은 다발성 골수종이다. 또 다른 양태에서, 암은 소세포 폐암(SCLC)과 같은 폐암이다. 또 다른 양태에서, 암은 신경모세포종이다. 또 다른 양태에서, 암은 버킷 림프종이다. 또 다른 양태에서, 암은 자궁경부암이다. 또 다른 양태에서, 암은 두경부암이다. 또 다른 양태에서, 암은 식도암이다. 또 다른 양태에서, 암은 위암이다. 또 다른 양태에서, 암은 췌장암이다. 또 다른 양태에서, 암은 간암이다. 또 다른 양태에서, 암은 흑색종이다. 또 다른 양태에서, 암은 난소암이다. 또 다른 양태에서, 암은 대장암이다. 또 다른 양태에서, 암은 전립선암이다. 또 다른 양태에서, 암은 유방암이다.
또 다른 양태에서, 본 발명에 따른 암은 급성 단핵구성 백혈병, 급성 골수성 백혈병, 만성 골수성 백혈병, 만성 림프구성 백혈병, 혼합 계통 백혈병, 림프종, NUT-중간선 암종, 다발성 골수종, 소세포 폐암, 비소세포 폐암, 신경모세포종, 버킷 림프종, 자궁경부암, 두경부암, 식도암, 위암, 췌장암, 간암, 흑색종, 난소암, 대장암, 전립선암, 유방암, 방광암, 신경교종, 육종, 식도 편평 세포 암종 및 유두 갑상선 암종으로 이루어진 군으로부터 선택된다.
즉, 본 발명은 화학식 1의 화합물 또는 이의 약학적으로 허용 가능한 염의 상기 질병을 치료 또는 예방하기 위한 의약 용도를 제공한다.
2. 개체 (Subjects)
본 발명에 따라 치료될 적합한 개체는 포유동물 개체를 포함한다. 본 발명에 따른 포유동물은, 이에 한정되는 것은 아니지만, 인간, 개(canine), 고양잇과동물(feline), 소(bovine), 염소(caprine), 말(equine), 양(ovine), 돼지(porcine), 설치류(rodents), 토끼목(lagomorphs), 영장류(primates) 등을 포함하고, 자궁 내의(in utero) 포유동물을 포함한다.
일 양태에서, 본 발명에 따른 치료될 적합한 개체는 인간이다.
3. 투여 및 투여량 (Administration and Dosing)
본 발명의 화합물은 일반적으로 치료적으로 유효한 양이 투여된다.
본 명세서에서 사용된 "유효량"은 SHP2 관련 질환의 진행을 늦추거나 또는 최소화하거나, SHP2 관련 질환의 치료 또는 관리에서 치료상 이점을 제공하기에 충분한 본 발명의 화합물의 양을 말한다. "유효량"은 또한 생체외(in vitro) 또는 생체내(in vivo) 어떤 쪽이든 SHP2 활성을 억제 또는 줄이기에 충분한 양을 말한다.
본 발명의 화합물은 임의의 적합한 경로에 의하여 이러한 경로에 적당한 약학 조성물의 형태, 그리고 의도된 치료를 위하여 효과적인 투여량으로 투여될 수 있다. 효과적인 투여량은 단일 또는 분할 투여로 일반적으로 약 0.001 내지 약 100 mg/체중kg/일이고, 바람직하게는 약 0.01 내지 약 50 mg/kg/일이다. 나이, 종, 및 치료될 질병 또는 상태(condition)에 따라 이 범위의 하한 미만의 투여량 수준이 적합할 수 있다. 다른 경우에는, 여전히 더 큰 투여량이 해로운 부작용없이 사용될 수 있다. 더 큰 투여량은 하루 동안 투여를 위하여, 여러 작은 투여량으로 분할될 수 있다.
본 발명 화합물의 약학 조성물
다른 양태에서, 본 발명은 화학식 1의 화합물 또는 이의 약학적으로 허용 가능한 염, 및 약학적으로 허용 가능한 담체 또는 첨가제를 포함하는 약학 조성물이 제공한다. 본 발명의 일 양태에 있어, 상기 약학 조성물의 용도는 후술하는 SHP2 관련 질환, 바람직하게는 암의 치료 또는 예방 용도이다.
용어 "약학적으로 허용 가능한"은 약학적 제제로 사용하기에 적합한 것을 의미하며, 일반적으로 이러한 사용을 위하여 안전한 것으로 간주되며, 이러한 사용을 위하여 국가의 관리 기관에 의하여 공식적으로 승인되거나 한국 약전 또는 미국 약전의 명단에 있는 것을 의미한다.
약학 조성물, 제형 및 투여 경로
상기 설명된 질병 또는 상태(condition)의 치료를 위하여, 본 명세서에서 설명된 상기 화합물 또는 이의 약학적으로 허용 가능한 염은 다음과 같이 투여될 수 있다.
구강 투여(Oral administration)
본 발명의 화합물은 구강으로 투여될 수 있으며, 구강은 연하(swallowing)를 포함하는 개념이다. 구강 투여에 의하여 본 발명의 화합물이 위장관(gastrointestinal tract)에 들어가거나, 예를 들어, 구강(buccal) 또는 설하(sublingual) 투여와 같이, 입으로부터 혈류로 직접적으로 흡수될 수 있다.
구강 투여를 위한 적합한 조성물은 고형상, 액상, 겔(gel), 또는 파우더 형상일 수 있으며, 정제(tablet), 로젠지(lozenge), 캡슐(capsule), 과립제, 산제 등의 제형을 가질 수 있다.
구강 투여를 위한 조성물은 선택적으로 장용 코팅(enteric coating)될 수 있으며, 장용 코팅을 통하여 지연된(delayed) 또는 지속된(sustained) 방출을 나타낼 수 있다. 즉, 본 발명에 따른 구강 투여를 위한 조성물은 즉시 또는 변형된(modified) 방출 패턴을 가진 제형일 수 있다.
액체 제형은 용액, 시럽 및 현탁액을 포함할 수 있으며, 이러한 액상 조성물은 연질 또는 경질 캡슐 내에 함유된 형태일 수 있다. 이러한 제형은 약학적으로 허용 가능한 담체, 예를 들어, 물, 에탄올, 폴리에틸렌글리콜, 셀룰로오스, 또는 오일(oil)을 포함할 수 있다. 상기 제형은 또한 하나 이상의 유화제 및/또는 현탁제를 포함할 수 있다.
정제(tablet) 제형에서, 활성 성분인 약물의 양은 정제 총 중량 대비 약 0.05 중량% 내지 약 95 중량%, 더욱 일반적으로 제형의 약 2 중량% 내지 약 50 중량%로 존재할 수 있다. 또한, 정제는 약 0.5 중량% 내지 약 35 중량%, 더욱 일반적으로 제형의 약 2 중량% 내지 약 25 중량%를 포함하는 붕해제를 함유할 수 있다. 붕해제의 예로는 유당, 전분, 소디움스타치글리콜레이트, 크로스포비돈, 크로스카멜로스소디움(croscarmellose sodium), 말토덱스트린 또는 이들의 혼합물이 사용될 수 있으나 이에 한정되는 것은 아니다.
정제로 제조하기 위해 포함되는 적합한 활택제는 약 0.1 중량% 내지 약 5 중량% 양으로 존재할 수 있고, 탈크(talc), 이산화규소, 스테아린산, 칼슘, 아연 또는 마그네슘 스테아레이트, 소듐 스테아릴 푸마레이트 등이 활택제로 사용될 수 있으나, 본 발명은 이러한 첨가제들의 종류에 한정되는 것은 아니다.
정제로 제조하기 위한 결합제(binder)로는 젤라틴, 폴리에틸렌글리콜, 당(sugar), 검(gum), 녹말(starch), 폴리비닐피롤리돈, 하이드록시프로필셀룰로오스, 하이드록시프로필메틸셀룰로오스 등이 사용될 수 있으며, 정제로 제조하기 위한 적합한 희석제로는 만니톨, 자일리톨, 락토오스, 덱스트로오스, 수크로오스, 솔비톨, 녹말(starch), 미결정셀룰로오스 등이 사용될 수 있으나, 본 발명은 이러한 첨가제들의 종류에 한정되는 것은 아니다.
선택적으로 정제에 포함될 수 있는 가용화제는 정제 총 중량 대비 약 0.1 중량% 내지 약 3 중량% 양이 사용될 수 있고, 예를 들어, 폴리소르베이트, 소디움 라우릴설페이트, 소디움 도데실설페이트, 프로필렌 카보네이트, 디에틸렌글리콜모노에틸에테르, 디메틸이소소르비드, 폴리옥시에틸렌글리콜화된 천연 또는 수소화 피마자유, HCORTM(Nikkol), 올레일에스테르, 젤루시어(GelucireTM), 카프릴릭/카프릴산 모노/디글리세리드, 소르비탄지방산에스테르, 솔루톨HSTM 등이 본 발명에 따른 약학 조성물에 사용될 수 있으나, 본 발명은 이러한 가용화제의 구체적 종류에 한정되는 것은 아니다.
비경구 투여(Parenteral Administration)
본 발명의 화합물은 혈류, 근육, 또는 내장 내로 직접 투여될 수 있다. 비경구 투여를 위한 적합한 방법은 정맥내(intravenous), 근육내(intra-muscular), 피하 동맥내(subcutaneous intraarterial), 복강내(intraperitoneal), 척추강내(intrathecal), 두개내(intracranial) 주사 등을 포함한다. 비경구 투여를 위한 적합한 장치는 (바늘 및 바늘 없는 주사기를 포함하는) 주사기(injector) 및 주입 방법(infusion method)을 포함한다.
비경구 투여를 위한 조성물은 즉시 또는 변형된 방출 패턴을 가진 제형일 수 있으며, 변형된 방출 패턴은 지연된(delayed) 또는 지속된(sustained) 방출 패턴일 수 있다.
대부분의 비경구 제형은 액상 조성물이며, 이러한 액상 조성물은 본 발명에 따른 약효 성분, 염, 완충제, 등장화제 등을 포함하는 수용액이다.
비경구 제형은 또한 건조된 형태(예를 들어, 동결 건조) 또는 멸균 비-수용액으로서 제조될 수 있다. 이들 제형은 멸균수(sterile water)와 같은 적합한 비히클(vehicle)과 함께 사용될 수 있다. 용해도 증강제(solubility-enhancing agents) 또한 비경구 용액의 제조에 사용될 수 있다.
국소 투여(Topical Administration)
본 발명의 화합물은 피부 또는 경피로 국소적으로 투여될 수 있다. 이 국소 투여를 위한 제형은 로션, 용액, 크림, 젤, 하이드로젤, 연고, 폼(foam), 임플란트(implant), 패치 등을 포함한다. 국소 투여 제형을 위한 약학적으로 허용 가능한 담체는 물, 알코올, 미네랄 오일, 글리세린, 폴리에틸렌글리콜 등을 포함할 수 있다. 국소 투여는 또한 전기천공법(electroporation), 이온도입법(iontophoresis), 음파영동(phonophoresis) 등에 의하여 수행될 수 있다.
국소 투여를 위한 조성물은 즉시 또는 변형된 방출 패턴을 가진 제형일 수 있으며, 변형된 방출 패턴은 지연된(delayed) 또는 지속된(sustained) 방출 패턴일 수 있다.
본 개시는 SHP2 분해활성이 매우 우수하여 다양한 약리 활성을 나타낼 수 있는 화합물, 이들을 유효 성분으로 포함하는 약학 조성물, 이들의 의약 용도(특히, 암 또는 종양) 및 이들을 치료 또는 예방이 필요한 개체에게 투여하는 것을 포함하는 치료 방법을 제공한다. 본 발명에 따른 화합물 또는 이들의 약학적으로 허용 가능한 염은 SHP2 분해활성, (대사)안정성, 물리화학적 성질 등의 다양한 측면에서 우수하며, 특히 유사한 구조를 가진 화합물들 대비 SHP2 분해활성이 훨씬 뛰어나다.
이하, 본 발명의 이해를 돕기 위하여 실시예 등을 들어 상세하게 설명하기로 한다. 그러나, 본 발명에 따른 실시예들은 여러 가지 다른 형태로 변형될 수 있으며, 본 발명의 범위가 하기 실시예들에 한정되는 것으로 해석되어서는 안 된다. 본 발명의 실시예들은 본 발명이 속한 분야에서 평균적인 지식을 가진 자에게 본 발명을 보다 완전하게 설명하기 위해 제공되는 것이다.
본 발명 화합물들의 제조
이하, 본 발명 일부 화합물들의 합성 과정을 기재하며, 하기 언급되지 않은 화합물들의 경우 출발 물질, 중간체 및/또는 반응 물질을 대체하여 유사한 방법으로 제조될 수 있다.
중간체 1. Tert-butyl (1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)carbamate
Figure PCTKR2022011797-appb-img-000011
단계 1: 3-(tert-butylthio)-2-chloroaniline의 합성
2-chloro-3-fluoroaniline (3 g, 20.60 mmol), 2-methylpropane-2-thiol (7 ml, 61.81 mmol), 및 cesium carbonate (13 g, 41.20 mmol)를 DMF (40 ml)에 현탁시킨 후 120 ℃에서 16시간 동안 교반하였다. 반응액을 EtOAc (400 ml)로 희석한 뒤 증류수 (150 ml x 3) 및 brine (150 ml x 3)으로 씻었다. 유기층을 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 갈색 오일 4.44 g (quant.)을 수득하였다.
단계 2: 3-amino-2-chlorobenzenethiol의 합성
3-(tert-butylthio)-2-chloroaniline (4.44 g, 20.58 mmol)을 conc. HCl (38 ml)에 현탁시킨 후 90 ℃에서 6시간 동안 교반하였다. 반응액을 냉각시킨 후 생성된 고체를 여과, 건조하여 상아색 고체 2.39 g (73%)을 수득하였다.
단계 3: 2-chloro-3-((5-chloropyrazin-2-yl)thio)aniline의 합성
3-amino-2-chlorobenzenethiol (1.34 g, 8.39 mmol), 2-bromo-5-chloropyrazine (1.6 g, 8.39 mmol), Pd2(dba)3 (77 mg, 0.08 mmol), Xantphos (97 mg, 0.17 mmol), 및 DIPEA (2.9 mg, 16.79 mmol)를 1,4-dioxane (15 ml)에 현탁시킨 후 95 ℃에서 1시간 동안 교반하였다. 반응액을 여과한 후 여과액을 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (3-15% EtOAc/Hexane)하여 상아색 고체 1.24 g (54%)을 수득하였다.
단계 4: tert-butyl (1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)carbamate의 합성
2-chloro-3-((5-chloropyrazin-2-yl)thio)aniline (120 mg, 0.44 mmol), tert-butyl (4-methylpiperidin-4-yl)carbamate (113 mg, 0.53 mmol), 및 DIPEA (0.35 ml, 1.99 mmol)를 NMP (0.48 ml)에 현탁시킨 후 130 ℃에서 16시간 동안 교반하였다. 반응액에 증류수 (15 ml)를 가한 뒤 EtOAc (15 ml)로 추출하고 유기층을 brine (15 ml)으로 씻었다. 이후 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-33% EtOAc/Hexane)하여 상아색 고체 175 mg (88%)를 수득하였다.
중간체 2. Tert-butyl (1-(6-amino-5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)carbamate
Figure PCTKR2022011797-appb-img-000012
2-bromo-5-chloropyrazine 대신 3-bromo-6-chloropyrazin-2-amine을 사용하여 중간체 1의 합성법과 동일한 방법으로 중간체 2를 합성하였다.
중간체 3. Tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate
Figure PCTKR2022011797-appb-img-000013
Tert-butyl (4-methylpiperidin-4-yl)carbamate 대신 tert-butyl ((4-methylpiperidin-4-yl)methyl)carbamate를 사용하여 중간체 1의 합성법과 동일한 방법으로 중간체 3을 합성하였다.
중간체 4. Tert-butyl ((1-(6-amino-5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate
Figure PCTKR2022011797-appb-img-000014
2-bromo-5-chloropyrazine 대신 3-bromo-6-chloropyrazin-2-amine을 사용하고, tert-butyl (4-methylpiperidin-4-yl)carbamate 대신 tert-butyl ((4-methylpiperidin-4-yl)methyl)carbamate를 사용하여 중간체 1의 합성법과 동일한 방법으로 중간체 4를 합성하였다.
중간체 5. Tert-butyl ((1-(5-((2-chloro-3-(methylamino)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate
Figure PCTKR2022011797-appb-img-000015
tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3, 100 mg, 0.22 mmol), 및 37% formaldehyde (87 μl, 1.08 mmol)를 DCM (2 ml)에 현탁시켰다. 이후 acetic acid (66 μl, 0.86 mmol), 및 sodium cyanoborohydride (27 mg, 0.43 mmol)를 가하고 상온에서 1시간 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (10 ml)을 가하고 DCM (10 ml x 2)로 추출하였다. 유기층을 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (1-30% EtOAc/Hexane)하여 상아색 고체 65.9 mg (64%)을 수득하였다.
중간체 6. Tert-butyl (R)-((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)pyrrolidin-3-yl)methyl)carbamate
Figure PCTKR2022011797-appb-img-000016
tert-butyl (4-methylpiperidin-4-yl)carbamate 대신 tert-butyl (S)-(pyrrolidin-3-ylmethyl)carbamate를 사용하여 중간체 1의 합성법과 동일한 방법으로 중간체 6을 합성하였다.
중간체 7. (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000017
단계 1: (2S,4R)-methyl 1-((S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxylate의 합성
(2S,4R)-methyl 4-hydroxypyrrolidine-2-carboxylate hydrochloride (7.85 g 43.23 mmol), (S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanoic acid (10 g, 43.23 mmol), EDCI (16.57 g, 86.46 mmol), HOBt (7 g, 86.46 mmol), 및 DIPEA (45 ml, 259.38 mmol)를 DMF (216 ml)에 현탁시킨 후 상온에서 16시간 동안 교반하였다. 반응액을 NH4Cl 포화수용액 (300 ml), 및 NaHCO3 포화수용액 (300 ml)로 씻은 후 EtOAc (150 ml x 2)로 추출하였다. 유기층을 brine (150 ml x 2)으로 씻고 무수 황산 나트륨으로 건조하였다. 이후 여과, 감압 농축하여 얻어진 잔사를 MPLC (0-5% MeOH/DCM)하여 상아색 고체 11 g (71%)을 수득하였다.
단계 2: (2S,4R)-1-((S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxylic acid의 합성
(2S,4R)-methyl 1-((S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxylate (781 mg, 2.18 mmol), 및 lithium hydroxide (366 mg, 8.72 mmol)를 water : THF = 1 : 1 (10 ml)에 현탁시킨 후 상온에서 3 시간 동안 교반하였다. 반응액을 1N HCl 수용액을 이용하여 pH 1-2로 조정한 뒤 EtOAc (15 ml x 2)로 추출하였다. 이후 유기층을 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 흰색 고체 680 mg (91%)를 수득하였다.
단계 3: 2-hydroxy-4-(4-methylthiazol-5-yl)benzonitrile의 합성
4-bromo-2-hydroxybenzonitrile (2 g, 10.15 mmol), 및 4-methylthiazole (1.83 ml, 20.20 mmol)을 DMAC (40 ml)에 현탁시킨 후 potassium acetate (1.98 g, 20.20 mmol), 및 Pd(OAc)2 (45 mg, 0.20 mmol)을 가하고 16시간 동안 가열 환류 하였다. 반응액에 증류수 (100 ml)를 가한 뒤 EtOAc (50 ml x 2)로 추출하였다. 유기층을 brine (50 ml x 2)로 씻고 무수 황산 마그네슘으로 건조하였다. 이후 여과, 감압 농축하여 얻어진 잔사를 MPLC (0-50% EtOAc/Hexane)하여 상아색 고체 980 mg (45%)을 수득하였다.
단계 4: 2-(aminomethyl)-5-(4-methylthiazol-5-yl)phenol의 합성
2-hydroxy-4-(4-methylthiazol-5-yl)benzonitrile (864 mg, 3.99 mmol)를 THF (22 ml)에 현탁시켰다. 이후 LiAlH4 (2.0 M in THF, 8 ml, 15.98 mmol)를 0 ℃에서 서서히 가하고 3 시간 동안 가열 환류 하였다. 반응액의 온도를 0 ℃로 낮춘 후 증류수 (0.5 ml)를 천천히 가하고 Rochelle solution (20 ml)로 중화하였다. 그 다음 celite filter로 여과하였다. 여과액을 MPLC (DCM : MeOH : NH4OH = 15 : 1 : 0.1)하여 노란색 고체 416 mg (47%)을 수득하였다.
단계 5: tert-butyl ((S)-1-((2S,4R)-4-hydroxy-2-((2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)carbamate의 합성
2-(aminomethyl)-5-(4-methylthiazol-5-yl)phenol (416 mg 1.89 mmol), (2S,4R)-1-((S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxylic acid (650 mg, 1.89 mmol), HATU (789 mg, 2.08 mmol), 및 DIPEA (0.98 ml, 5.66 mmol)를 DMF (6 ml)에 현탁시킨 후 상온에서 16시간 동안 교반하였다. 반응액에 증류수 (25 ml)로 가한 후 EtOAc (15 ml x 2)로 추출하였다. 유기층을 brine (15 ml x 2)로 씻고 무수 황산 나트륨으로 건조하였다. 이후 여과, 감압 농축하여 얻어진 잔사를 MPLC (0-20% MeOH/DCM)하여 연한 노란색 고체 898 mg (87%)을 수득하였다.
단계 6: (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide의 합성
tert-butyl ((S)-1-((2S,4R)-4-hydroxy-2-((2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)carbamate (885 mg, 1.62 mmol)를 DCM (30 ml)에 현탁시킨 후 4 M HCl in dioxane (8 ml)를 가하고 상온에서 2 시간 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (20 ml)을 가하여 중화한 뒤 DCM (10 ml x 2)로 추출하였다. 유기층을 brine (10 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 얻어진 잔사를 MPLC (amine-silica, 0-10% MeOH/DCM)하여 노란색 고체 522 mg (72%)을 수득하였다.
단계 7: (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide의 합성
(2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (399 mg 0.89 mmol), 1-fluorocyclopropane-1-carboxylic acid (92.6 mg, 0.89 mmol), HATU (381 mg, 0.89 mmol), 및 DIPEA (0.8 ml, 4.47 mmol)를 DMF (4 ml)에 현탁시킨 후 상온에서 16시간 동안 교반하였다. 반응액에 증류수 (10 ml)를 가하고 EtOAc (11 ml x 2)로 추출하였다. 유기층을 brine (10 ml x 2)로 씻고 무수황산나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-10% MeOH/DCM)하여 흰색 고체 377 mg (79%)을 수득하였다.
중간체 8. (2S,4R)-1-((S)-2-(cyclopropanecarboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000018
1-fluorocyclopropane-1-carboxylic acid 대신 cyclopropanecarboxylic acid를 사용하여 중간체 7의 합성법과 동일한 방법으로 중간체 8을 합성하였다.
중간체 9. (2S,4R)-1-((S)-2-acetamido-3,3-dimethylbutanoyl)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000019
1-fluorocyclopropane-1-carboxylic acid 대신 acetic acid를 사용하여 중간체 7의 합성법과 동일한 방법으로 중간체 9를 합성하였다.
중간체 10. (2S,4R)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000020
(2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (중간체 7, 350 mg, 0.657 mmol), 및 1-bromo-2-chloroethane (0.2 ml, 2.628 mmol)을 DMF (2 ml)에 현탁시킨 후 potassium carbonate (272 mg, 1.971 mmol), 및 potassium iodide (218 mg, 1.314 mmol)을 가하고 70 ℃에서 16시간 동안 교반하였다. 반응액에 증류수 (20 ml)를 가한 뒤 EtOAc (20 ml x 2)로 추출하였다. 유기층을 brine (20 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-5% MeOH/DCM)하여 상아색 고체 234 mg (60%)을 수득하였다.
중간체 10의 합성법과 동일한 방법으로 하기 표 2의 중간체 11-18을 합성하였다.
Figure PCTKR2022011797-appb-img-000021
Figure PCTKR2022011797-appb-img-000022
실시예 1. N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-1-(3-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)propyl)piperidine-4-carboxamide
Figure PCTKR2022011797-appb-img-000023
단계 1: 1-(((9H-fluoren-9-yl)methoxy)carbonyl)piperidine-4-carboxylic acid의 합성
4-piperidinecarboxylic acid (1 g, 7.74 mmol), 및 Na2CO3 (2.5 g, 23.22 mmol)를 water : 1,4-dioxane = 1 : 1 (64 ml)에 현탁시킨 후 0 ℃에서 10분 동안 교반하였다. Fmoc-Cl (2 g, 7.74 mmol)를 가한 후 상온에서 16시간 동안 교반하였다. 반응액을 1N HCl 수용액을 이용하여 pH = 6으로 조정한 뒤 EtOAc (30 ml x 2)로 추출하였다. 유기층을 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-10% MeOH/DCM)하여 흰색 고체 1.91 g (70%)를 수득하였다.
단계2: (9H-fluoren-9-yl)methyl 4-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)carbamoyl)piperidine-1-carboxylate의 합성
1-(((9H-fluoren-9-yl)methoxy)carbonyl)piperidine-4-carboxylic acid (758 mg, 2.16 mmol)를 DCM (40 ml)에 현탁시킨 후 oxalyl chloride (0.24 ml, 2.80 mmol), 및 DMF (3 drops)를 가하고 상온에서 2.5 시간 동안 교반하였다. 반응액을 농축하여 노란색 오일을 얻었다. 얻은 노란색 오일을 DCM (15 ml)에 녹인 후 tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3, 500 mg, 1.08 mmol), 및 DIPEA (0.75 ml, 4.31 mmol)를 가하고 상온에서 1시간 동안 교반하였다. 반응액에 증류수 (20 ml)를 가한 뒤 DCM (15 ml x 2)로 추출하였다. 얻어진 유기층을 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-5% MeOH/DCM)하여 상아색 고체 908 mg (quant.)를 수득하였다.
단계 3: tert-butyl ((1-(5-((2-chloro-3-(piperidine-4-carboxamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate의 합성
(9H-fluoren-9-yl)methyl 4-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)carbamoyl)piperidine-1-carboxylate (908 mg, 1.14 mmol)을 DMF (7.2 ml)에 녹인 후 piperidine (1.9 ml, 19.36 mmol)을 천천히 가하고 상온에서 1시간 동안 교반하였다. 반응액에 증류수 (20 ml)를 가한 뒤 EtOAc (20 ml x 2)로 추출하고 유기층을 무수 황산 나트륨으로 건조하였다. 이후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (amine-silica, 0-100% EtOAc/Hexane, 0-5% MeOH/DCM)하여 상아색 고체 462 mg (71%)을 수득하였다.
단계 4: tert-butyl ((1-(5-((2-chloro-3-(1-(3-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)propyl)piperidine-4-carboxamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate의 합성
tert-butyl ((1-(5-((2-chloro-3-(piperidine-4-carboxamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (16 mg, 0.03 mmol), 및 (2S,4R)-N-(2-(3-chloropropoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 11, 18.6 mg, 0.03 mmol)을 DMF (0.04 ml)에 현탁시킨 후 potassium carbonate (12 mg, 0.08 mmol), 및 potassium iodide (9 mg, 0.06 mmol)을 가하고 70 ℃에서 16시간 동안 교반하였다. 반응액에 증류수 (1 ml)를 가한 뒤 EtOAc (2 ml x 2)로 추출하였다. 유기층을 brine (1 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-10% MeOH/DCM)하여 상아색 고체 20.8 mg (65%)을 수득하였다.
단계 5: N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-1-(3-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)propyl)piperidine-4-carboxamide의 합성
tert-butyl ((1-(5-((2-chloro-3-(1-(3-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)propyl)piperidine-4-carboxamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (20.8 mg, 0.02 mmol)를 DCM (0.9 ml)에 현탁시킨 후 4 M HCl in dioxane (0.04 ml)를 가하고 상온에서 1 시간 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (10 ml)를 가하여 중화한 뒤 DCM (5 ml x 2)로 추출하였다. 유기층을 brine (5 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (amine-silica, 0-5% MeOH/DCM)하여 상아색 고체 15.4 mg (81%)을 수득하였다.
실시예 2. N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-1-(4-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)butyl)piperidine-4-carboxamide
Figure PCTKR2022011797-appb-img-000024
(2S,4R)-N-(2-(3-chloropropoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 11) 대신 (2S,4R)-N-(2-(4-chlorobutoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 12)를 사용하여 실시예 1의 합성법과 동일한 방법으로 실시예 2를 합성하였다.
실시예 3. N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-1-(6-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)hexyl)piperidine-4-carboxamide
Figure PCTKR2022011797-appb-img-000025
(2S,4R)-N-(2-(3-chloropropoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 11) 대신 (2S,4R)-N-(2-((6-chlorohexyl)oxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 13)를 사용하여 실시예 1의 합성법과 동일한 방법으로 실시예 3을 합성하였다.
실시예 4. N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-1-(8-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)octyl)piperidine-4-carboxamide
Figure PCTKR2022011797-appb-img-000026
(2S,4R)-N-(2-(3-chloropropoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 11) 대신 (2S,4R)-N-(2-((8-chlorooctyl)oxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 14)를 사용하여 실시예 1의 합성법과 동일한 방법으로 실시예 4를 합성하였다.
실시예 5. (2S,4R)-N-(2-(2-(4-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)piperidin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000027
Piperidine-4-carboxylic acid 대신 2-(piperidin-4-yl)acetic acid를 사용하고, (2S,4R)-N-(2-(3-chloropropoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 11) 대신 (2S,4R)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 10)를 사용하여 실시예 1의 합성법과 동일한 방법으로 실시예 5를 합성하였다.
실시예 6. (2S,4R)-N-(2-(3-(4-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)piperidin-1-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000028
piperidine-4-carboxylic acid 대신 2-(piperidin-4-yl)acetic acid를 사용하여 실시예 1의 합성법과 동일한 방법으로 실시예 6을 합성하였다.
실시예 7. (2S,4R)-N-(2-(4-(4-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)piperidin-1-yl)butoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000029
Piperidine-4-carboxylic acid 대신 2-(piperidin-4-yl)acetic acid를 사용하고, (2S,4R)-N-(2-(3-chloropropoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 11) 대신 (2S,4R)-N-(2-(4-chlorobutoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 12)를 사용하여 실시예 1의 합성법과 동일한 방법으로 실시예 7을 합성하였다.
실시예 8. (2S,4R)-N-(2-(3-(4-(2-((3-((5-(4-amino-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)piperidin-1-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000030
Piperidine-4-carboxylic acid 대신 2-(piperidin-4-yl)acetic acid를 사용하고, tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3) 대신 tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 1)를 사용하여 실시예 1의 합성법과 동일한 방법으로 실시예 8을 합성하였다.
실시예 9. (2S,4R)-N-(2-(3-(4-(2-((3-((3-amino-5-(4-amino-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)piperidin-1-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000031
Piperidine-4-carboxylic acid 대신 2-(piperidin-4-yl)acetic acid를 사용하고, tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3) 대신 tert-butyl (1-(6-amino-5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)carbamate (중간체 2)를 사용하여 실시예 1의 합성법과 동일한 방법으로 실시예 9를 합성하였다.
실시예 10. (2S,4R)-N-(2-(3-(4-(2-((3-((3-amino-5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)piperidin-1-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000032
Piperidine-4-carboxylic acid 대신 2-(piperidin-4-yl)acetic acid를 사용하고, tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3) 대신 tert-butyl ((1-(6-amino-5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 4)를 사용하여 실시예 1의 합성법과 동일한 방법으로 실시예 10을 합성하였다.
실시예 11. (2S,4R)-N-(2-(3-(4-(2-((3-((5-((R)-3-(aminomethyl)pyrrolidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)piperidin-1-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000033
Piperidine-4-carboxylic acid 대신 2-(piperidin-4-yl)acetic acid를 사용하고, tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3) 대신 tert-butyl (R)-((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)pyrrolidin-3-yl)methyl)carbamate (중간체 6)를 사용하여 실시예 1의 합성법과 동일한 방법으로 실시예 11을 합성하였다.
실시예 12. (2S,4R)-1-((S)-2-acetamido-3,3-dimethylbutanoyl)-N-(2-(3-(4-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)piperidin-1-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000034
Piperidine-4-carboxylic acid 대신 2-(piperidin-4-yl)acetic acid를 사용하고, (2S,4R)-N-(2-(3-chloropropoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 11) 대신 (2S,4R)-1-((S)-2-acetamido-3,3-dimethylbutanoyl)-N-(2-(3-chloropropoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 18)를 사용하여 실시예 1의 합성법과 동일한 방법으로 실시예 12를 합성하였다.
실시예 13. (2S,4R)-N-(2-(2-(4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropyl)piperidin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000035
Piperidine-4-carboxylic acid 대신 3-(piperidin-4-yl)propanoic acid를 사용하고, (2S,4R)-N-(2-(3-chloropropoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 11) 대신 (2S,4R)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 10)을 사용하여 실시예 1의 합성법과 동일한 방법으로 실시예 13을 합성하였다.
실시예 14. (2S,4R)-N-(2-(3-(4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropyl)piperidin-1-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000036
Piperidine-4-carboxylic acid 대신 3-(piperidin-4-yl)propanoic acid를 사용하여 실시예 1의 합성법과 동일한 방법으로 실시예 14를 합성하였다.
실시예 15. (2S,4R)-N-(2-(4-(4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropyl)piperidin-1-yl)butoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000037
Piperidine-4-carboxylic acid 대신 3-(piperidin-4-yl)propanoic acid를 사용하고, (2S,4R)-N-(2-(3-chloropropoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 11) 대신 (2S,4R)-N-(2-(4-chlorobutoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 12)를 사용하여 실시예 1의 합성법과 동일한 방법으로 실시예 15를 합성하였다.
실시예 16. (2S,4R)-N-(2-(2-(4-(4-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-4-oxobutyl)piperidin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000038
Piperidine-4-carboxylic acid 대신 4-(piperidin-4-yl)butanoic acid를 사용하고, (2S,4R)-N-(2-(3-chloropropoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 11) 대신 (2S,4R)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 10)을 사용하여 실시예 1의 합성법과 동일한 방법으로 실시예 16을 합성하였다.
실시예 17. (2S,4R)-N-(2-(4-(4-(4-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-4-oxobutyl)piperidin-1-yl)butoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000039
Piperidine-4-carboxylic acid 대신 4-(piperidin-4-yl)butanoic acid를 사용하고, (2S,4R)-N-(2-(3-chloropropoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 11) 대신 (2S,4R)-N-(2-(4-chlorobutoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 12)를 사용하여 실시예 1의 합성법과 동일한 방법으로 실시예 17을 합성하였다.
실시예 18. (2S,4R)-N-(2-((6-(4-(4-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-4-oxobutyl)piperidin-1-yl)hexyl)oxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000040
Piperidine-4-carboxylic acid 대신 4-(piperidin-4-yl)butanoic acid를 사용하고, (2S,4R)-N-(2-(3-chloropropoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 11) 대신 (2S,4R)-N-(2-((6-chlorohexyl)oxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 13)를 사용하여 실시예 1의 합성법과 동일한 방법으로 실시예 18을 합성하였다.
실시예 19. (2S,4R)-N-(2-((6-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)carbamoyl)azetidin-1-yl)hexyl)oxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000041
Piperidine-4-carboxylic acid 대신 azetidine-3-carboxylic acid를 사용하고, (2S,4R)-N-(2-(3-chloropropoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 11) 대신 (2S,4R)-N-(2-((6-chlorohexyl)oxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 13)를 사용하여 실시예 1의 합성법과 동일한 방법으로 실시예 19를 합성하였다.
실시예 20. ((2S,4R)-N-(2-(3-(4-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)-4-hydroxypiperidin-1-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000042
단계 1: 2-(1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-hydroxypiperidin-4-yl)acetic acid의 합성
2-(4-hydroxypiperidin-4-yl)acetic acid (38 mg, 0.19 mmol), Na2CO3 (61 mg, 0.58 mmol)를 water : 1,4-dioxane = 1 : 1 (1.6 ml)에 현탁시킨 후 0 ℃에서 10분 동안 교반하였다. Fmoc-Cl (50 mg, 0.19 mmol)를 가한 후 상온에서 16시간 동안 교반하였다. 반응액을 1N HCl 수용액을 이용하여 pH = 3으로 조정한 뒤 EtOAc (10 ml)로 추출하였다. 유기층을 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-3% MeOH/DCM)하여 흰색 고체 48.6 mg (66%)를 수득하였다.
단계 2: (9H-fluoren-9-yl)methyl 4-(2-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)-4-hydroxypiperidine-1-carboxylate의 합성
2-(1-(((9H-fluoren-9-yl)methoxy)carbonyl)-4-hydroxypiperidin-4-yl)acetic acid (25 mg, 0.06 mmol), tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3, 30 mg, 0.06 mmol), TCFH (108 mg, 0.38 mmol), 및 NMI (0.051 ml, 0.65 mmol)를 ACN : THF = 1 : 1 (0.5 ml)에 현탁시킨 후 상온에서 5 시간 동안 교반하였다. 반응액에 증류수 (7 ml)를 가한 뒤 EtOAc (10 ml)로 추출하였다. 유기층을 brine (7 ml)로 씻고 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (3-50% EtOAc/Hexane)하여 상아색 고체 11.7 mg (23%)을 수득하였다.
단계 3: tert-butyl ((1-(5-((2-chloro-3-(2-(4-hydroxypiperidin-4-yl)acetamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate의 합성
(9H-fluoren-9-yl)methyl 4-(2-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)-4-hydroxypiperidine-1-carboxylate (40 mg, 0.04 mmol)을 DMF (0.2 ml)에 녹인 후 piperidine (0.01 ml, 0.97 mmol)을 천천히 가하고 상온에서 2시간 동안 교반하였다. 반응액에 증류수 (7 ml)를 가한 뒤 EtOAc (10 ml)로 추출하였다. 유기층을 brine (7 ml)로 씻고 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (amine silica, 0-4% MeOH/DCM)하여 상아색 고체 9.7 mg (37%)을 수득하였다.
단계 4: tert-butyl ((1-(5-((2-chloro-3-(2-(1-(3-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)propyl)-4-hydroxypiperidin-4-yl)acetamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate의 합성
tert-butyl ((1-(5-((2-chloro-3-(2-(4-hydroxypiperidin-4-yl)acetamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (9.7 mg, 0.02 mmol), 및 (2S,4R)-N-(2-(3-chloropropoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 11, 12 mg, 0.02 mmol)를 DMF (0.05 ml)에 현탁시킨 후 potassium carbonate (6.6 mg, 0.05 mmol)를 가하고 70 ℃에서 16시간 동안 교반하였다. 반응액에 증류수 (3 ml)를 가한 뒤 EtOAc (5 ml)로 추출하였다. 유기층을 brine (3 ml)로 씻고 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-10% MeOH/DCM)하여 상아색 고체 8.1 mg (43%)을 수득하였다.
단계 5: ((2S,4R)-N-(2-(3-(4-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)-4-hydroxypiperidin-1-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide의 합성
tert-butyl ((1-(5-((2-chloro-3-(2-(1-(3-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)propyl)-4-hydroxypiperidin-4-yl)acetamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (8.1 mg, 0.007 mmol)를 DCM (0.7 ml)에 현탁시킨 후 4 M HCl in dioxane (0.017 ml)를 가하고 상온에서 1 시간 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (5 ml)를 가하여 중화한 뒤 DCM (10 ml x 2)로 추출하였다. 유기층을 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (amine-silica, 0-3% MeOH/DCM)하여 상아색 고체 5.8 mg (78%)을 수득하였다.
실시예 21. N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-1-(4-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)butanoyl)piperidine-4-carboxamide
Figure PCTKR2022011797-appb-img-000043
단계 1: ethyl 4-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)butanoate의 합성
(2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (중간체 7, 97 mg, 0.18 mmol), 및 ethyl 4-bromobutyrate (0.05 ml, 0.36 mmol)을 DMF (0.3 ml)에 현탁시킨 후 potassium carbonate (75 mg, 0.55 mmol)를 가하고 70 ℃에서 16시간 동안 교반하였다. 반응액에 증류수 (10 ml)를 가한 뒤 EtOAc (10 ml x 2)로 추출하였다. 유기층을 brine (10 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-10% MeOH/DCM)하여 상아색 고체 104 mg (88%)을 수득하였다.
단계 2: 4-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)butanoic acid의 합성
ethyl 4-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)butanoate (102 mg, 0.16 mmol), 및 lithium hydroxide (16 mg, 0.37 mmol)를 water : THF = 2 : 1 (1.8 ml)에 현탁시킨 후 상온에서 4시간 동안 교반하였다. 반응액을 1N HCl 수용액을 이용하여 pH = 2로 조정한 뒤 EtOAc (10 ml x 2)로 추출하였다. 유기층을 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 상아색 고체 60 mg (61%)를 수득하였다.
단계 3: tert-butyl ((1-(5-((2-chloro-3-(1-(4-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)butanoyl)piperidine-4-carboxamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate의 합성
4-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)butanoic acid (19 mg 0.03 mmol), tert-butyl ((1-(5-((2-chloro-3-(piperidine-4-carboxamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (18 mg, 0.03 mmol), EDCI (7.2 mg, 0.04 mmol), HOBt (5 mg, 0.04 mmol), 및 DIPEA (0.02 mL, 0.13 mmol)를 DMF (0.5 ml)에 현탁시킨 후 상온에서 16시간 동안 교반하였다. 반응액에 증류수 (2 ml)를 가한 뒤 EtOAc (2 ml x 2)로 추출하였다. 유기층을 brine (2 ml x 2)로 씻고 유기층을 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (10% MeOH/DCM)하여 상아색 고체 12.2 mg (33%)을 수득하였다.
단계 4: N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-1-(4-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)butanoyl)piperidine-4-carboxamide의 합성
tert-butyl ((1-(5-((2-chloro-3-(1-(4-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)butanoyl)piperidine-4-carboxamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (12.2 mg, 0.01 mmol)를 DCM (0.5 ml)에 현탁시킨 후 4 M HCl in dioxane (0.03 ml)를 가하고 상온에서 1 시간 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (2 ml)을 가한 뒤 10% MeOH/DCM (5 ml x 2)로 추출하였다. 유기층을 brine (5 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 잔사를 MPLC (amine-silica, 0-5% MeOH/DCM)하여 상아색 고체 7.8 mg (70%)을 수득하였다.
실시예 22. (2S,4R)-N-(2-(2-(4-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)piperidin-1-yl)-2-oxoethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000044
Ethyl 4-bromobutyrate 대신 ethyl 2-bromoacetate를 사용하고, tert-butyl ((1-(5-((2-chloro-3-(piperidine-4-carboxamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate 대신 tert-butyl ((1-(5-((2-chloro-3-(2-(piperidin-4-yl)acetamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate를 사용하여 실시예 21의 합성법과 동일한 방법으로 실시예 22를 합성하였다.
실시예 23. (2S,4R)-N-(2-(4-(4-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)piperidin-1-yl)-4-oxobutoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000045
Tert-butyl ((1-(5-((2-chloro-3-(piperidine-4-carboxamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate 대신 tert-butyl ((1-(5-((2-chloro-3-(2-(piperidin-4-yl)acetamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate를 사용하여 실시예 21의 합성법과 동일한 방법으로 실시예 23을 합성하였다.
실시예 24. (2S,4R)-N-(2-(4-(4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropyl)piperidin-1-yl)-4-oxobutoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000046
Tert-butyl ((1-(5-((2-chloro-3-(piperidine-4-carboxamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate 대신 tert-butyl ((1-(5-((2-chloro-3-(3-(piperidin-4-yl)propanamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate를 사용하여 실시예 21의 합성법과 동일한 방법으로 실시예 24를 합성하였다.
실시예 25. (2S,4R)-N-(2-(4-(4-(4-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-4-oxobutyl)piperidin-1-yl)-4-oxobutoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000047
Tert-butyl ((1-(5-((2-chloro-3-(piperidine-4-carboxamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate 대신 tert-butyl ((1-(5-((2-chloro-3-(4-(piperidin-4-yl)butanamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)mthyl)carbamate를 사용하여 실시예 21의 합성법과 동일한 방법으로 실시예 25를 합성하였다.
실시예 26. (2S,4R)-N-(2-((6-(4-(4-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-4-oxobutyl)piperidin-1-yl)-6-oxohexyl)oxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000048
Ethyl 4-bromobutyrate 대신 methyl 6-bromohexanoate를 사용하고, tert-butyl ((1-(5-((2-chloro-3-(piperidine-4-carboxamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate 대신 tert-butyl ((1-(5-((2-chloro-3-(4-(piperidin-4-yl)butanamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate를 사용하여 실시예 21의 합성법과 동일한 방법으로 실시예 26을 합성하였다.
실시예 27. (2S,4R)-N-(2-(2-(1-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoacetyl)piperidin-4-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000049
단계 1: ethyl 2-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoacetate의 합성
Tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3, 100 mg, 0.22 mmol)를 DCM (2 ml)에 현탁시킨 후 DCM (1 ml)에 녹인 ethyl chloroglyoxylate (27.9 mg, 0.20 mmol)를 0 ℃에서 천천히 가하고 30분 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (2 ml)을 가하고 10% MeOH/DCM (20 ml x 2)로 추출하였다. 유기층을 brine (5 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 잔사를 MPLC (amine-silica, 30-40% EtOAc/Hexane)하여 상아색 고체 116 mg (95%)을 수득하였다.
단계 2: 2-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoacetic acid의 합성
Ethyl 2-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoacetate (116 mg, 0.21 mmol), lithium hydroxide (17 mg, 0.41 mmol)를 THF : MeOH : H2O = 3 : 2 : 1 (0.74 ml)에 현탁시킨 후 상온에서 2시간 동안 교반하였다. 반응액을 1N HCl 수용액을 이용하여 pH = 3으로 조정한 뒤 EtOAc (10 ml x 2)로 추출하였다. 유기층을 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 상아색 고체 100 mg (91%)를 수득하였다.
단계 3: tert-butyl 4-(2-hydroxyethyl)piperidine-1-carboxylate의 합성
2-(piperidin-4-yl)ethan-1-ol (1 g, 7.74 mmol), triethylamine (3.2 ml, 23.22 mmol)를 chloroform (16 ml)에 현탁시킨 후 0 ℃에서 di-tert-butyl dicarbonate (1.9 ml, 8.05 mmol)를 천천히 가하고 1시간 동안 교반하였다. 반응액에 NH4Cl 포화수용액 (45 ml)을 가하고 DCM (20 ml x 2)로 추출하였다. 유기층을 brine (20 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 무색 오일 2.12 g (quant.)을 수득하였다.
단계 4: tert-butyl 4-(2-(tosyloxy)ethyl)piperidine-1-carboxylate의 합성
Tert-butyl 4-(2-hydroxyethyl)piperidine-1-carboxylate (2.1 g, 9.11 mmol), triethylamine (1.4 ml, 10.02 mmol)를 DCM (13 ml)에 현탁시킨 후 4-toluenesulfonyl chloride (1.74 g, 9.11 mmol)를 가하고 상온에서 16시간 동안 교반하였다. 반응액에 증류수 (10 ml)를 가한 뒤 DCM (10 ml x 2)로 추출하였다. 유기층을 brine (10 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 잔사를 MPLC (0-50% EtOAc/Hexane)하여 무색 오일 2.41 g (69%)을 수득하였다.
단계 5: tert-butyl 4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperidine-1-carboxylate의 합성
(2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (중간체 7, 261 mg, 0.49 mmol), 및 tert-butyl 4-(2-(tosyloxy)ethyl)piperidine-1-carboxylate (376 mg, 0.98 mmol)을 DMF (3.2 ml)에 현탁시킨 후 potassium carbonate (203 mg, 1.47 mmol), 및 potassium iodide (163 mg, 0.98 mmol)를 가하고 70 ℃에서 16시간 동안 교반하였다. 반응액에 증류수 (10 ml)를 가한 뒤 EtOAc (10 ml x 2)로 추출하였다. 유기층을 brine (10 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-10% MeOH/DCM)하여 연한 노란색 고체 246 mg (68%)을 수득하였다.
단계 6: (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)-2-(2-(piperidin-4-yl)ethoxy)benzyl)pyrrolidine-2-carboxamide의 합성
tert-butyl 4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperidine-1-carboxylate (245 mg, 0.33 mmol)를 DCM (3.3 ml)에 현탁시킨 후 4 M HCl in dioxane (1.65 ml)를 가하고 상온에서 1 시간 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (10 ml)을 가하고 DCM (5 ml x 2)로 추출하였다. 유기층을 brine (5 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 노란색 고체 129 mg (61%)을 수득하였다.
단계 7: tert-butyl ((1-(5-((2-chloro-3-(2-(4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperidin-1-yl)-2-oxoacetamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate의 합성
2-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoacetic acid (25 mg 0.05 mmol), (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)-2-(2-(piperidin-4-yl)ethoxy)benzyl)pyrrolidine-2-carboxamide (30 mg, 0.05 mmol), EDCI (11 mg, 0.06 mmol), HOBt (8 mg, 0.06 mmol), 및 DIPEA (0.03 ml, 0.18 mmol)를 DMF (0.75 ml)에 현탁시킨 후 상온에서 16시간 동안 교반하였다. 반응액에 증류수 (3 ml)를 가한 뒤 EtOAc (3 ml x 2)로 추출하였다. 유기층을 brine (3 ml x 2)로 씻고 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 얻어진 잔사를 MPLC (0-7% MeOH/DCM)하여 상아색 고체 6.9 mg (13%)을 수득하였다.
단계 8: (2S,4R)-N-(2-(2-(1-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoacetyl)piperidin-4-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide의 합성
tert-butyl ((1-(5-((2-chloro-3-(2-(4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperidin-1-yl)-2-oxoacetamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (6.9 mg, 0.005 mmol)를 DCM (0.6 ml)에 현탁시킨 후 4 M HCl in dioxane (0.014 ml)를 가하고 상온에서 2 시간 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (1 ml)을 가하고 10% MeOH/DCM (5 ml x 2)로 추출하였다. 유기층을 brine (1 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (amine-silica, 0-4% MeOH/DCM)하여 상아색 고체 4.2 mg (67%)을 수득하였다.
실시예 28. (2S,4R)-N-(2-(3-(1-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoacetyl)piperidin-4-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000050
2-(piperidin-4-yl)ethan-1-ol 대신 3-(piperidin-4-yl)propan-1-ol를 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 28을 합성하였다.
실시예 29. (2S,4R)-N-(2-((1-(3-((3-((5-(4-amino-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperidin-4-yl)methoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000051
Tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3) 대신 tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 1)를 사용하고, ethyl chloroglyoxylate 대신 ethyl 3-chloro-3-oxopropanoate를 사용하고, 2-(piperidin-4-yl)ethan-1-ol 대신 piperidin-4-ylmethanol를 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 29를 합성하였다.
실시예 30. (2S,4R)-N-(2-((1-(3-((3-((3-amino-5-(4-amino-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperidin-4-yl)methoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000052
tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3) 대신 tert-butyl (1-(6-amino-5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)carbamate (중간체 2)를 사용하고, ethyl chloroglyoxylate 대신 ethyl 3-chloro-3-oxopropanoate를 사용하고, 2-(piperidin-4-yl)ethan-1-ol 대신 piperidin-4-ylmethanol를 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 30을 합성하였다.
실시예 31. (2S,4R)-N-(2-((1-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperidin-4-yl)methoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000053
ethyl chloroglyoxylate 대신 ethyl 3-chloro-3-oxopropanoate를 사용하고, 2-(piperidin-4-yl)ethan-1-ol 대신 piperidin-4-ylmethanol를 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 31을 합성하였다.
실시예 32. (2S,4R)-N-(2-((1-(3-((3-((3-amino-5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperidin-4-yl)methoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000054
tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3) 대신 tert-butyl ((1-(6-amino-5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 4)를 사용하고, ethyl chloroglyoxylate 대신 ethyl 3-chloro-3-oxopropanoate를 사용하고, 2-(piperidin-4-yl)ethan-1-ol 대신 piperidin-4-ylmethanol를 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 32를 합성하였다.
실시예 33. (2S,4R)-1-((S)-2-acetamido-3,3-dimethylbutanoyl)-N-(2-((1-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperidin-4-yl)methoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000055
ethyl chloroglyoxylate 대신 ethyl 3-chloro-3-oxopropanoate를 사용하고, 2-(piperidin-4-yl)ethan-1-ol 대신 piperidin-4-ylmethanol를 사용하고, (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (중간체 7) 대신 (2S,4R)-1-((S)-2-acetamido-3,3-dimethylbutanoyl)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (중간체 9)를 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 33을 합성하였다.
실시예 34. (2S,4R)-N-(2-(2-(1-(3-((3-((5-(4-amino-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperidin-4-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000056
tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3) 대신 tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 1)를 사용하고, ethyl chloroglyoxylate 대신 ethyl 3-chloro-3-oxopropanoate를 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 34를 합성하였다.
실시예 35. (2S,4R)-N-(2-(2-(1-(3-((3-((3-amino-5-(4-amino-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperidin-4-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000057
tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3) 대신 tert-butyl (1-(6-amino-5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)carbamate (중간체 2)를 사용하고, ethyl chloroglyoxylate 대신 ethyl 3-chloro-3-oxopropanoate를 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 35를 합성하였다.
실시예 36. (2S,4R)-N-(2-(2-(1-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperidin-4-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000058
ethyl chloroglyoxylate 대신 ethyl 3-chloro-3-oxopropanoate를 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 36을 합성하였다.
실시예 37. (2S,4R)-N-(2-(2-(1-(3-((3-((3-amino-5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperidin-4-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000059
tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3) 대신 tert-butyl ((1-(6-amino-5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 4)를 사용하고, ethyl chloroglyoxylate 대신 ethyl 3-chloro-3-oxopropanoate를 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 37을 합성하였다.
실시예 38. (2S,4R)-1-((S)-2-acetamido-3,3-dimethylbutanoyl)-N-(2-(2-(1-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperidin-4-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000060
ethyl chloroglyoxylate 대신 ethyl 3-chloro-3-oxopropanoate를 사용하고, (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (중간체 7) 대신 (2S,4R)-1-((S)-2-acetamido-3,3-dimethylbutanoyl)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (중간체 9)를 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 38을 합성하였다.
실시예 39. (2S,4R)-N-(2-((1-(3-((3-((5-(4-amino-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperidin-4-yl)methoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000061
tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3) 대신 tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 1)를 사용하고, ethyl chloroglyoxylate 대신 ethyl 3-chloro-3-oxopropanoate를 사용하고, 2-(piperidin-4-yl)ethan-1-ol 대신 3-(piperidin-4-yl)propan-1-ol를 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 39를 합성하였다.
실시예 40. (2S,4R)-N-(2-(3-(1-(3-((3-((3-amino-5-(4-amino-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperidin-4-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000062
tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3) 대신 tert-butyl (1-(6-amino-5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)carbamate (중간체 2)를 사용하고, ethyl chloroglyoxylate 대신 ethyl 3-chloro-3-oxopropanoate를 사용하고, 2-(piperidin-4-yl)ethan-1-ol 대신 3-(piperidin-4-yl)propan-1-ol를 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 40을 합성하였다.
실시예 41. (2S,4R)-N-(2-(3-(1-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperidin-4-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000063
ethyl chloroglyoxylate 대신 ethyl 3-chloro-3-oxopropanoate를 사용하고, 2-(piperidin-4-yl)ethan-1-ol 대신 3-(piperidin-4-yl)propan-1-ol를 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 41을 합성하였다.
실시예 42. (2S,4R)-N-(2-(3-(1-(3-((3-((3-amino-5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperidin-4-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000064
tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3) 대신 tert-butyl ((1-(6-amino-5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 4)를 사용하고, ethyl chloroglyoxylate 대신 ethyl 3-chloro-3-oxopropanoate를 사용하고, 2-(piperidin-4-yl)ethan-1-ol 대신 3-(piperidin-4-yl)propan-1-ol를 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 42를 합성하였다.
실시예 43. (2S,4R)-1-((S)-2-acetamido-3,3-dimethylbutanoyl)-N-(2-(3-(1-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperidin-4-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000065
ethyl chloroglyoxylate 대신 ethyl 3-chloro-3-oxopropanoate를 사용하고, 2-(piperidin-4-yl)ethan-1-ol 대신 3-(piperidin-4-yl)propan-1-ol를 사용하고, (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (중간체 7) 대신 (2S,4R)-1-((S)-2-acetamido-3,3-dimethylbutanoyl)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (중간체 9)를 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 43을 합성하였다.
실시예 44. (2S,4R)-N-(2-((1-(4-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-4-oxobutanoyl)piperidin-4-yl)methoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000066
ethyl chloroglyoxylate 대신 methyl 4-chloro-4-oxobutanoate를 사용하고, 2-(piperidin-4-yl)ethan-1-ol 대신 piperidin-4-ylmethanol를 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 44를 합성하였다.
실시예 45. (2S,4R)-N-(2-(2-(1-(4-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-4-oxobutanoyl)piperidin-4-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000067
ethyl chloroglyoxylate 대신 methyl 4-chloro-4-oxobutanoate를 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 45를 합성하였다.
실시예 46. (2S,4R)-N-(2-((1-(1-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)carbamoyl)cyclopropane-1-carbonyl)piperidin-4-yl)methoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000068
ethyl chloroglyoxylate 대신 methyl 1-(chlorocarbonyl)cyclopropane-1-carboxylate를 사용하고, 2-(piperidin-4-yl)ethan-1-ol 대신 piperidin-4-ylmethanol를 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 46을 합성하였다.
실시예 47. (2S,4R)-N-(2-(2-(1-(1-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)carbamoyl)cyclopropane-1-carbonyl)piperidin-4-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000069
ethyl chloroglyoxylate 대신 methyl 1-(chlorocarbonyl)cyclopropane-1-carboxylate를 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 47을 합성하였다.
실시예 48. (2S,4R)-N-(2-(3-(1-(1-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)carbamoyl)cyclopropane-1-carbonyl)piperidin-4-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000070
ethyl chloroglyoxylate 대신 methyl 1-(chlorocarbonyl)cyclopropane-1-carboxylate를 사용하고, 2-(piperidin-4-yl)ethan-1-ol 대신 3-(piperidin-4-yl)propan-1-ol을 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 48을 합성하였다.
실시예 49. (2S,4R)-N-(2-(2-(1-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2,2-dimethyl-3-oxopropanoyl)piperidin-4-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000071
ethyl chloroglyoxylate 대신 methyl 3-chloro-2,2-dimethyl-3-oxopropanoate를 사용하여 실시예 27의 합성법과 동일한 방법으로 실시예 49를 합성하였다.
실시예 50. (2S,4R)-N-(2-((7-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)-7-azaspiro[3.5]nonan-2-yl)oxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000072
Figure PCTKR2022011797-appb-img-000073
단계 1: ethyl 3-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoate의 합성
tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3, 240 mg, 0.52 mmol)를 DCM (3.2 ml)에 현탁시킨 후 ethyl malonyl chloride (0.08 ml, 0.62 mmol)를 0 ℃에서 천천히 가하고 30분 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (10 ml)을 가하고 DCM (10 ml x 2)으로 추출하였다. 유기층을 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 얻어진 잔사를 MPLC (3-35% EtOAc/Hexane)하여 상아색 고체 206 mg (69%)을 수득하였다.
단계 2: 3-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoic acid의 합성
ethyl 3-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoate (50 mg, 0.09 mmol), lithium hydroxide (11 mg, 0.26 mmol)를 THF : MeOH : H2O = 3 : 2 : 1 (0.34 ml)에 현탁시킨 후 상온에서 2시간 동안 교반하였다. 반응액을 1N HCl 수용액을 이용하여 pH = 2로 조정한 뒤 EtOAc (10 ml x 2)로 추출하였다. 유기층을 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 상아색 고체 46 mg (97%)를 수득하였다.
단계 3: tert-butyl 2-(((2-nitrophenyl)sulfonyl)oxy)-7-azaspiro[3.5]nonane-7-carboxylate의 합성
tert-butyl 2-hydroxy-7-azaspiro[3.5]nonane-7-carboxylate (2 g, 8.29 mmol), triethylamine (2.3 ml, 16.58 mmol)를 DCM (16 ml)에 현탁시킨 후 2-nitrobenzenesulfonyl chloride (1.84 g, 8.29 mmol)를 가하고 상온에서 2시간 동안 교반하였다. 반응액에 증류수 (30 ml)를 가한 뒤 DCM (50 ml)로 추출하였다. 유기층을 brine (30 ml)로 씻고 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 얻어진 잔사를 MPLC (1-40% EtOAc/Hexane)하여 무색 오일 3.3 g (93%)을 수득하였다.
단계 4: tert-butyl 2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)-7-azaspiro[3.5]nonane-7-carboxylate의 합성
(2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (중간체 7, 500 mg, 0.494 mmol), tert-butyl 2-(((2-nitrophenyl)sulfonyl)oxy)-7-azaspiro[3.5]nonane-7-carboxylate (1.2 g, 2.82 mmol)을 DMF (1.5 ml)에 현탁시킨 후 cesium carbonate (612 mg, 1.88 mmol)를 가하고 50 ℃에서 16시간 동안 교반하였다. 반응액에 증류수 (20 ml)를 가한 뒤 EtOAc (30 ml)로 추출하였다. 유기층을 brine (20 ml)로 씻고 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 얻어진 잔사를 MPLC (0-4% MeOH/DCM)하여 상아색 고체 675 mg (95%)을 수득하였다.
단계 5: (2S,4R)-N-(2-((7-azaspiro[3.5]nonan-2-yl)oxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide의 합성
tert-butyl 2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)-7-azaspiro[3.5]nonane-7-carboxylate (675 mg, 0.89 mmol)를 DCM (18 ml)에 현탁시킨 후 4 M HCl in dioxane (4.4 ml)를 가하고 상온에서 1 시간 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (10 ml)을 가하고 DCM (15 ml x 2)로 추출하였다. 유기층을 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 상아색 고체 483 mg (82%)를 수득하였다.
단계 6: tert-butyl ((1-(5-((2-chloro-3-(3-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)-7-azaspiro[3.5]nonan-7-yl)-3-oxopropanamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate의 합성
3-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoic acid (16.5 mg 0.03 mmol), (2S,4R)-N-(2-((7-azaspiro[3.5]nonan-2-yl)oxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (19.4 mg, 0.03 mmol), EDCI (6.8 mg, 0.04 mmol), HOBt (4.8 mg, 0.04 mmol), DIPEA (0.02 ml, 0.12 mmol)를 DMF (0.1 ml)에 현탁시킨 후 상온에서 16시간 동안 교반하였다. 반응액에 증류수 (7 ml)를 가한 뒤 EtOAc (10 ml)로 추출하였다. 유기층을 brine (7 ml)로 씻고 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 얻어진 잔사를 MPLC (0-5% MeOH/DCM)하여 상아색 고체 20 mg (42%)을 수득하였다.
단계 7: (2S,4R)-N-(2-((7-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)-7-azaspiro[3.5]nonan-2-yl)oxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide의 합성
tert-butyl ((1-(5-((2-chloro-3-(3-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)-7-azaspiro[3.5]nonan-7-yl)-3-oxopropanamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (20 mg, 0.017 mmol)를 DCM (1.7 ml)에 현탁시킨 후 4 M HCl in dioxane (0.04 ml)를 가하고 상온에서 2 시간 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (7 ml)을 가하고 DCM (10 ml x 3)로 추출하였다. 유기층을 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 얻어진 잔사를 MPLC (amine-silica, 0-3% MeOH/DCM)하여 상아색 고체 16 mg (87%)을 수득하였다.
실시예 51. (2S,4R)-N-(2-((7-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)(methyl)amino)-3-oxopropanoyl)-7-azaspiro[3.5]nonan-2-yl)oxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000074
tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3) 대신 tert-butyl ((1-(5-((2-chloro-3-(methylamino)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 5)를 사용하여 실시예 50의 합성법과 동일한 방법으로 실시예 51을 합성하였다.
실시예 52. (2S,4R)-N-(2-(2-(4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperazin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000075
단계 1: ethyl 3-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoate의 합성
tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3, 240 mg, 0.52 mmol)를 DCM (3.2 ml)에 현탁시킨 후 ethyl malonyl chloride (0.08 ml, 0.62 mmol)를 0 ℃에서 천천히 가하고 30분 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (10 ml)을 가하고 DCM (10 ml x 2)으로 추출하였다. 유기층을 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (3-35% EtOAc/Hexane)하여 상아색 고체 206 mg (69%)을 수득하였다.
단계 2: 3-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoic acid의 합성
ethyl 3-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoate (50 mg, 0.09 mmol), lithium hydroxide (11 mg, 0.26 mmol)를 THF : MeOH : H2O = 3 : 2 : 1 (0.34 ml)에 현탁시킨 후 상온에서 2시간 동안 교반하였다. 반응액을 1N HCl 수용액을 이용하여 pH = 2로 조정한 뒤 EtOAc (10 ml x 2)로 추출하였다. 유기층을 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 상아색 고체 46 mg (97%)를 수득하였다.
단계 3. tert-butyl 4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-1-carboxylate의 합성
(2S,4R)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 10, 100 mg, 0.16 mmol), tert-butyl piperazine-1-carboxylate (38 mg, 0.2 mmol), potassium carbonate (70 mg, 0.5 mmol), 및 sodium iodide (26 mg, 0.16 mmol)를 acetonitrile (1 ml)에 현탁시킨 후 60 ℃에서 16시간 동안 교반하였다. 반응액에 증류수 (10 ml)를 가한 뒤 EtOAc (10 ml x 2)로 추출하였다. 유기층을 brine (10 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-10% MeOH/DCM)하여 흰색 고체 89 mg (72%)을 수득하였다.
단계 4: (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)-2-(2-(piperazin-1-yl)ethoxy)benzyl)pyrrolidine-2-carboxamide의 합성
tert-butyl 4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-1-carboxylate (86 mg, 0.12 mmol)를 DCM (1.15 ml)에 현탁시킨 후 4 M HCl in dioxane (0.58 ml)를 가하고 상온에서 1 시간 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (15 ml)을 가하고 10% MeOH/DCM (15 ml x 2)로 추출하였다. 유기층을 brine (15 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 상아색 고체 69 mg (93%)을 수득하였다.
단계 5: tert-butyl ((1-(5-((2-chloro-3-(3-(4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazin-1-yl)-3-oxopropanamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate의 합성
3-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoic acid (25 mg 0.05 mmol), (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)-2-(2-(piperazin-1-yl)ethoxy)benzyl)pyrrolidine-2-carboxamide (29 mg, 0.05 mmol), EDCI (10 mg, 0.05 mmol), HOBt (7 mg, 0.05 mmol), 및 DIPEA (0.02 ml, 0.02 mmol)를 DMF (0.15 ml)에 현탁시킨 후 상온에서 16시간 동안 교반하였다. 반응액에 증류수 (10 ml)를 가한 뒤 EtOAc (5 ml x 2)로 추출하였다. 유기층을 brine (5 ml x 2)로 씻고 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (10% MeOH/DCM)하여 상아색 고체 19.3 mg (36%)을 수득하였다.
단계 6: (2S,4R)-N-(2-(2-(4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperazin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide의 합성
tert-butyl ((1-(5-((2-chloro-3-(3-(4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazin-1-yl)-3-oxopropanamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (17.1 mg, 0.01 mmol)를 DCM (0.3 ml)에 현탁시킨 후 4 M HCl in dioxane (0.07 ml)를 가하고 상온에서 1 시간 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (10 ml)을 가하고 10% MeOH/DCM (5 ml x 2)로 추출하였다. 유기층을 brine (5 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (amine-silica, 0-5% MeOH/DCM)하여 흰색 고체 13.5 mg (86%)을 수득하였다.
실시예 53. (2S,4R)-N-(2-(2-(4-(3-((3-((5-(4-amino-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperazin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000076
tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3) 대신 tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 1)를 사용하여 실시예 52의 합성법과 동일한 방법으로 실시예 53을 합성하였다.
실시예 54. (2S,4R)-N-(2-(2-(4-(3-((3-((3-amino-5-(4-amino-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperazin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000077
tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3) 대신 tert-butyl (1-(6-amino-5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)carbamate (중간체 2)를 사용하여 실시예 52의 합성법과 동일한 방법으로 실시예 54를 합성하였다.
실시예 55. (2S,4R)-N-(2-(2-(4-(3-((3-((3-amino-5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperazin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000078
tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3) 대신 tert-butyl ((1-(6-amino-5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 4)를 사용하여 실시예 52의 합성법과 동일한 방법으로 실시예 55를 합성하였다.
실시예 56. (2S,4R)-N-(2-(2-(4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperazin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(cyclopropanecarboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000079
(2S,4R)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 10) 대신 (2S,4R)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(cyclopropanecarboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 15)를 사용하여 실시예 52의 합성법과 동일한 방법으로 실시예 56을 합성하였다.
실시예 57. (2S,4R)-1-((S)-2-acetamido-3,3-dimethylbutanoyl)-N-(2-(2-(4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperazin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000080
(2S,4R)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 10) 대신 ((2S,4R)-1-((S)-2-acetamido-3,3-dimethylbutanoyl)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 17)을 사용하여 실시예 52의 합성법과 동일한 방법으로 실시예 57을 합성하였다.
실시예 58. (2S,4R)-N-(2-(2-(4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-methyl-3-oxopropanoyl)piperazin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000081
Ethyl malonyl chloride대신 ethyl 3-chloro-2-methyl-3-oxopropanoate를 사용하여 실시예 52의 합성법과 동일한 방법으로 실시예 58을 합성하였다.
실시예 59. (2S,4R)-N-(2-(2-(4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2,2-dimethyl-3-oxopropanoyl)piperazin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000082
Ethyl malonyl chloride 대신 methyl 3-chloro-2,2-dimethyl-3-oxopropanoate를 사용하여 실시예 52의 합성법과 동일한 방법으로 실시예 59를 합성하였다.
실시예 60. (2S,4R)-N-(2-(3-(4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperazin-1-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000083
(2S,4R)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 10) 대신 (2S,4R)-N-(2-(3-chloropropoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 11)를 사용하여 실시예 52의 합성법과 동일한 방법으로 실시예 60을 합성하였다.
실시예 61. (2S,4R)-1-((S)-2-acetamido-3,3-dimethylbutanoyl)-N-(2-(3-(4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)piperazin-1-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000084
(2S,4R)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 10) 대신 (2S,4R)-1-((S)-2-acetamido-3,3-dimethylbutanoyl)-N-(2-(3-chloropropoxy)-4-(4-methylthiazol-5-yl)benzyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 18)를 사용하여 실시예 52의 합성법과 동일한 방법으로 실시예 61을 합성하였다.
실시예 62. (2S,4R)-N-(2-(2-((R)-4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)-3-methylpiperazin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000085
tert-butyl piperazine-1-carboxylate 대신 tert-butyl (R)-2-methylpiperazine-1-carboxylate를 사용하여 실시예 52의 합성법과 동일한 방법으로 실시예 62를 합성하였다.
실시예 63. (2S,4R)-N-(2-(2-((S)-4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)-3-methylpiperazin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000086
tert-butyl piperazine-1-carboxylate 대신 tert-butyl (S)-2-methylpiperazine-1-carboxylate를 사용하여 실시예 52의 합성법과 동일한 방법으로 실시예 63을 합성하였다.
실시예 64. (2S,4R)-N-(2-(3-((S)-4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)-3-methylpiperazin-1-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000087
(2S,4R)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 10) 대신 (2S,4R)-N-(2-(3-chloropropoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 11)를 사용하고, tert-butyl piperazine-1-carboxylate 대신 tert-butyl (S)-2-methylpiperazine-1-carboxylate를 사용하여 실시예 52의 합성법과 동일한 방법으로 실시예 64를 합성하였다.
실시예 65. ((2S,4R)-N-(2-(3-((R)-4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)-3-methylpiperazin-1-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000088
(2S,4R)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 10) 대신 (2S,4R)-N-(2-(3-chloropropoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 11)를 사용하고, tert-butyl piperazine-1-carboxylate 대신 tert-butyl (R)-2-methylpiperazine-1-carboxylate를 사용하여 실시예 52의 합성법과 동일한 방법으로 실시예 65를 합성하였다.
실시예 66. (S)-methyl 4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)-1-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-2-carboxylate
Figure PCTKR2022011797-appb-img-000089
Tert-butyl piperazine-1-carboxylate 대신 (S)-1-tert-butyl 3-methyl piperazine-1,3-dicarboxylate를 사용하여 실시예 52의 합성법과 동일한 방법으로 실시예 66을 합성하였다.
실시예 67. methyl (R)-4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)-1-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-2-carboxylate
Figure PCTKR2022011797-appb-img-000090
Tert-butyl piperazine-1-carboxylate 대신 (R)-1-(tert-butyl) 3-methyl piperazine-1,3-dicarboxylate를 사용하여 실시예 52의 합성법과 동일한 방법으로 실시예 67을 합성하였다.
실시예 68. (2S,4R)-N-(2-(2-(8-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000091
Tert-butyl piperazine-1-carboxylate 대신 tert-butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate를 사용하여 실시예 52의 합성법과 동일한 방법으로 실시예 68을 합성하였다.
실시예 69. (S)-4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)-1-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-2-carboxylic acid
Figure PCTKR2022011797-appb-img-000092
Figure PCTKR2022011797-appb-img-000093
단계 1. 1-(tert-butyl) 3-methyl (S)-4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-1,3-dicarboxylate의 합성
(2S,4R)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 10, 100 mg, 0.16 mmol), 1-(tert-butyl) 3-methyl (S)-piperazine-1,3-dicarboxylate (82 mg, 0.34 mmol), potassium carbonate (70 mg, 0.50 mmol), 및 sodium iodide (56 mg, 0.34 mmol)를 DMF (0.6 ml)에 현탁시킨 후 70 ℃에서 16시간 동안 교반하였다. 반응액에 증류수 (10 ml)를 가한 뒤 EtOAc (15 ml)로 추출하였다. 유기층을 brine (10 ml)로 씻고 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-4% MeOH/DCM)하여 흰색 고체 91 mg (67%)을 수득하였다.
단계 2: methyl (S)-1-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-2-carboxylate의 합성
1-(tert-butyl) 3-methyl (S)-4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-1,3-dicarboxylate (90.7 mg, 0.11 mmol)를 DCM (1.1 ml)에 현탁시킨 후 4 M HCl in dioxane (0.28 ml)를 가하고 상온에서 1 시간 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (10 ml)을 가하고 DCM (20 ml x 2)로 추출하였다. 유기층을 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (amine silica, 0-3% MeOH/DCM)하여 상아색 고체 64 mg (80%)을 수득하였다.
단계 3: (S)-1-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-2-carboxylic acid의 합성
methyl (S)-1-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-2-carboxylate (64 mg 0.05 mmol), 및 lithium hydroxide (8 mg, 0.18 mmol)를 THF : MeOH : H2O = 3 : 2 : 1 (0.65 ml)에 현탁시킨 후 상온에서 16시간 동안 교반하였다. 반응액을 1N HCl 수용액을 이용하여 pH = 2로 조정한 뒤 감압 농축하여 상아색 고체 82 mg (quant.)를 수득하였다.
단계 4: (S)-4-(3-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)-1-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-2-carboxylic acid의 합성
3-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoic acid (20 mg, 0.04 mmol), NHS (4.3 mg, 0.04 mmol), 및 DCC (8 mg, 0.04 mmol)를 THF (0.4 ml)에 현탁시킨 후 상온에서 30분 동안 교반하였다. 반응액에 (S)-1-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-2-carboxylic acid (33.6 mg, 0.04 mmol), DIPEA (0.03 ml, 0.15 mmol), 및 DMF (0.4 ml)를 가한 뒤 상온에서 16시간 동안 교반하였다. 반응액에 증류수 (5 ml)를 가한 뒤 DCM (10 ml x 2)로 추출하였다. 유기층을 brine (5 ml)로 씻고 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-15% MeOH/DCM -> C18 reversed phase silica, 10-50% ACN/water)하여 흰색 고체 16.2 mg (36%)을 수득하였다.
단계 5: (S)-4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)-1-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-2-carboxylic acid의 합성
(S)-4-(3-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)-1-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-2-carboxylic acid (14 mg, 0.01 mmol)를 DCM (1.1 ml)에 현탁시킨 후 4 M HCl in dioxane (0.03 ml)를 가하고 상온에서 2 시간 동안 교반하였다. 반응액을 감압 농축하고 진공 건조하여 상아색 고체 7.5 mg (55%)을 수득하였다.
실시예 70. (R)-4-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropanoyl)-1-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazine-2-carboxylic acid
Figure PCTKR2022011797-appb-img-000094
1-(tert-butyl) 3-methyl (S)-piperazine-1,3-dicarboxylate 대신 1-(tert-butyl) 3-methyl (R)-piperazine-1,3-dicarboxylate를 사용하여 실시예 69의 합성법과 동일한 방법으로 실시예 70을 합성하였다.
실시예 71. (2S,4R)-N-(2-(3-(1-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)piperidin-4-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000095
단계 1: tert-butyl ((1-(5-((3-(2-bromoacetamido)-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate의 합성
tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3, 20 mg, 0.04 mmol), triethylamine (0.018 ml, 0.13 mmol)를 DCM (0.2 ml)에 현탁시킨 후 0 ℃에서 bromoacetyl chloride (7.5 mg, 0.05 mmol)를 천천히 가하고 30분 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (2 ml)을 가하고 DCM (5 ml x 2)로 추출하였다. 유기층을 brine (5 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (5-40% EtOAc/Hexane)하여 상아색 고체 10 mg (40%)을 수득하였다.
단계 2: tert-butyl ((1-(5-((2-chloro-3-(2-(4-(3-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)propyl)piperidin-1-yl)acetamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate의 합성
Tert-butyl ((1-(5-((3-(2-bromoacetamido)-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (35 mg, 0.06 mmol), (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)-2-(3-(piperidin-4-yl)propoxy)benzyl)pyrrolidine-2-carboxamide (20 mg, 0.03 mmol), potassium carbonate (13 mg, 0.09 mmol), 및 potassium iodide (10 mg, 0.06 mmol)를 DMF (0.2 ml)에 현탁시킨 후 70 ℃에서 16시간 동안 교반하였다. 반응액에 증류수 (10 ml)를 가한 뒤 EtOAc (5 ml x 2)로 추출하였다. 유기층을 brine (5 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-10% MeOH/DCM)하여 흰색 고체 22.3 mg (63%)을 수득하였다.
단계 3: (2S,4R)-N-(2-(3-(1-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)piperidin-4-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide의 합성
Tert-butyl ((1-(5-((2-chloro-3-(2-(4-(3-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)propyl)piperidin-1-yl)acetamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (22.3 mg, 0.02 mmol)를 DCM (1.9 ml)에 현탁시킨 후 4 M HCl in dioxane (0.05 ml)를 가하고 상온에서 1 시간 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (1 ml)을 가하고 10% MeOH/DCM (2 ml x 2)로 추출하였다. 유기층을 brine (1 ml x 2)로 씻고 무수 황산 마그네슘로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (amine-silica, 0-5% MeOH/DCM)하여 흰색 고체 16.2 mg (79%)을 수득하였다.
실시예 72. (2S,4R)-N-(2-(2-(4-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)piperazin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000096
(2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)-2-(3-(piperidin-4-yl)propoxy)benzyl)pyrrolidine-2-carboxamide 대신 (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)-2-(2-(piperazin-1-yl)ethoxy)benzyl)pyrrolidine-2-carboxamide를 사용하여 실시예 71의 합성법과 동일한 방법으로 실시예 72를 합성하였다.
실시예 73. (2S,4R)-N-(2-(3-(4-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)piperazin-1-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000097
(2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)-2-(3-(piperidin-4-yl)propoxy)benzyl)pyrrolidine-2-carboxamide 대신 (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)-2-(3-(piperazin-1-yl)propoxy)benzyl)pyrrolidine-2-carboxamide를 사용하여 실시예 71의 합성법과 동일한 방법으로 실시예 73을 합성하였다.
실시예 74. (2S,4R)-N-(2-(2-(1-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropyl)piperidin-4-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000098
단계 1: tert-butyl ((1-(5-((3-acrylamido-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate의 합성
tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3, 50 mg, 0.11 mmol), triethylamine (0.090 ml, 0.65 mmol)을 DCM (2 ml)에 현탁시킨 후 0 ℃에서 3-bromopropanoyl chloride (65 mg, 0.38 mmol)를 천천히 가하고 1시간 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (10 ml)을 가하고 DCM (20 ml)로 추출하였다. 유기층을 brine (10 ml)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (5-40% EtOAc/Hexane)하여 상아색 고체 36 mg (64%)을 수득하였다.
단계 2: tert-butyl ((1-(5-((2-chloro-3-(3-(4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperidin-1-yl)propanamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate의 합성
tert-butyl ((1-(5-((3-acrylamido-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (18 mg, 0.03 mmol), (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)-2-(2-(piperidin-4-yl)ethoxy)benzyl)pyrrolidine-2-carboxamide (25 mg, 0.04 mmol), 및 triethylamine (0.01 ml, 0.07 mmol)을 DCM : MeOH = 1 : 1 (0.2 ml)에 현탁시킨 후 60 ℃에서 16시간 동안 교반하였다. 반응액에 증류수 (3 ml)를 가한 뒤 DCM (5 ml)로 추출하고 유기층을 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-10% MeOH/DCM)하여 상아색 고체 21.3 mg (53%)을 수득하였다.
단계 3: (2S,4R)-N-(2-(2-(1-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropyl)piperidin-4-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide의 합성
tert-butyl ((1-(5-((2-chloro-3-(3-(4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperidin-1-yl)propanamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (21.3 mg, 0.02 mmol)를 DCM (1.8 ml)에 현탁시킨 후 4 M HCl in dioxane (0.05 ml)를 가하고 상온에서 1 시간 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (1 ml)을 가하고 DCM (5 ml x 3)로 추출하였다. 유기층을 brine (1 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (amine-silica, 0-4% MeOH/DCM)하여 흰색 고체 16.3 mg (84%)을 수득하였다.
실시예 75. (2S,4R)-N-(2-(3-(1-(3-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-3-oxopropyl)piperidin-4-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000099
(2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)-2-(2-(piperidin-4-yl)ethoxy)benzyl)pyrrolidine-2-carboxamide 대신 (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)-2-(3-(piperidin-4-yl)propoxy)benzyl)pyrrolidine-2-carboxamide를 사용하여 실시예 74의 합성법과 동일한 방법으로 실시예 75를 합성하였다.
실시예 76. (2S,4R)-N-(2-(2-(4-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)-3-oxopiperazin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000100
Figure PCTKR2022011797-appb-img-000101
단계 1: tert-butyl 4-(2-(tert-butoxy)-2-oxoethyl)-3-oxopiperazine-1-carboxylate의 합성
tert-butyl 3-oxopiperazine-1-carboxylate (200 mg, 1.00 mmol)을 DMF (1 ml)에 현탁시킨 후 0 ℃에서 sodium hydride (48 mg, 1.20 mmol)를 천천히 가하고 10분 동안 교반하였다. 반응액에 tert-butyl bromoacetate (0.16 ml, 1.10 mmol)를 가하고 1시간 동안 교반하였다. 반응액에 증류수 (10 ml)를 가하고 EtOAc (15 ml)로 추출하였다. 유기층을 brine (10 ml)로 씻고 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (3-50% EtOAc/Hexane)하여 무색 오일 286 mg (91%)을 수득하였다.
단계 2: 2-(2-oxopiperazin-1-yl)acetic acid의 합성
tert-butyl 4-(2-(tert-butoxy)-2-oxoethyl)-3-oxopiperazine-1-carboxylate (286 mg, 0.91 mmol)를 DCM (3 ml)에 현탁시킨 후 4 M HCl in dioxane (4.5 ml)를 가하고 상온에서 4 시간 동안 교반하였다. 반응액을 감압 농축하여 흰색 고체 173 mg (98%)을 수득하였다.
단계3: 2-(4-(((9H-fluoren-9-yl)methoxy)carbonyl)-2-oxopiperazin-1-yl)acetic acid의 합성
2-(2-oxopiperazin-1-yl)acetic acid (173 mg, 0.89 mmol), Na2CO3 (282 mg, 2.67 mmol)를 water : 1,4-dioxane = 1 : 1 (7.4 ml)에 현탁시킨 후 0 ℃에서 10분 동안 교반하였다. Fmoc-Cl (230 mg, 0.89 mmol)를 가한 후 상온에서 16시간 동안 교반하였다. 반응액을 1N HCl 수용액을 이용하여 pH = 3으로 조정한 뒤 EtOAc (15 ml)로 추출하였다. 유기층을 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-5% MeOH/DCM)하여 흰색 고체 278 mg (82%)를 수득하였다.
단계4: (9H-fluoren-9-yl)methyl 4-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)carbamoyl)piperidine-1-carboxylate의 합성
2-(4-(((9H-fluoren-9-yl)methoxy)carbonyl)-2-oxopiperazin-1-yl)acetic acid (80 mg, 0.22 mmol), tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3, 85 mg, 0.17 mmol), TCFH (145 mg, 0.52 mmol), NMI (0.068 ml, 0.86 mmol)를 ACN : THF = 1 : 1 (1.2 ml)에 현탁시킨 후 상온에서 16 시간 동안 교반하였다. 반응액에 증류수 (10 ml)를 가한 뒤 EtOAc (15 ml)로 추출하였다. 유기층을 brine (10 ml)로 씻고 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (3-70% EtOAc/Hexane)하여 상아색 고체 156 mg (quant.)을 수득하였다.
단계 5: tert-butyl ((1-(5-((2-chloro-3-(2-(2-oxopiperazin-1-yl)acetamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate의 합성
((9H-fluoren-9-yl)methyl 4-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)carbamoyl)piperidine-1-carboxylate (156 mg, 0.17 mmol)을 DMF (0.8 ml)에 녹인 후 piperidine (0.34 ml, 3.45 mmol)을 천천히 가하고 상온에서 2시간 동안 교반하였다. 반응액에 증류수 (10 ml)를 가한 뒤 EtOAc (20 ml)로 추출하였다. 유기층을 brine (10 ml x 2)로 씻고 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (amine-silica, 0-4 % MeOH/DCM)하여 상아색 고체 64.5 mg (57%)을 수득하였다.
단계 6: tert-butyl ((1-(5-((2-chloro-3-(2-(4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)-2-oxopiperazin-1-yl)acetamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate의 합성
Tert-butyl ((1-(5-((2-chloro-3-(2-(2-oxopiperazin-1-yl)acetamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (20 mg, 0.03 mmol), 및 (2S,4R)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 10, 25 mg, 0.04 mmol)을 DMF (0.01 ml)에 현탁시킨 후 potassium carbonate (14 mg, 0.10 mmol), potassium iodide (11 mg, 0.07 mmol)을 가하고 70 ℃에서 16시간 동안 교반하였다. 반응액에 증류수 (7 ml)를 가한 뒤 EtOAc (10 ml)로 추출하였다. 유기층을 brine (7 ml x 2)로 씻고 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-5% MeOH/DCM)하여 상아색 고체 27 mg (70%)을 수득하였다.
단계 7: (2S,4R)-N-(2-(2-(4-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)-3-oxopiperazin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide의 합성
Tert-butyl ((1-(5-((2-chloro-3-(2-(4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)-2-oxopiperazin-1-yl)acetamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (27 mg, 0.02 mmol)를 DCM (2.3 ml)에 현탁시킨 후 4 M HCl in dioxane (0.06 ml)를 가하고 상온에서 1 시간 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (5 ml)를 가하여 중화한 뒤 DCM (10 ml x 2)로 추출하였다. 유기층을 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (amine-silica, 0-3% MeOH/DCM)하여 상아색 고체 15 mg (61%)을 수득하였다.
실시예 77. (2S,4R)-N-(2-(3-(4-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)-3-oxopiperazin-1-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000102
(2S,4R)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 10) 대신 (2S,4R)-N-(2-(3-chloropropoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 11)를 사용하여 실시예 76의 합성법과 동일한 방법으로 실시예 77을 합성하였다.
실시예 78. (2S,4R)-N-(2-(4-(4-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)-3-oxopiperazin-1-yl)butoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000103
(2S,4R)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 10) 대신 (2S,4R)-N-(2-(4-chlorobutoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 12)를 사용하여 실시예 76의 합성법과 동일한 방법으로 실시예 78을 합성하였다.
실시예 79. (2S,4R)-N-(2-(2-(4-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)-3-oxopiperazin-1-yl)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(cyclopropanecarboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000104
(2S,4R)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 10) 대신 (2S,4R)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(cyclopropanecarboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 15)를 사용하여 실시예 76의 합성법과 동일한 방법으로 실시예 79를 합성하였다.
실시예 80. (2S,4R)-N-(2-(3-(4-(2-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-2-oxoethyl)-3-oxopiperazin-1-yl)propoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(cyclopropanecarboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000105
(2S,4R)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 10) 대신 (2S,4R)-N-(2-(3-chloropropoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(cyclopropanecarboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 16)을 사용하여 실시예 76의 합성법과 동일한 방법으로 실시예 80을 합성하였다.
실시예 81. N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-5-(4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazin-1-yl)picolinamide
Figure PCTKR2022011797-appb-img-000106
단계 1: 5-fluoropicolinic acid의 합성
Methyl 5-fluoropicolinate (500 mg, 3.22 mmol)를 THF (5 ml)에 현탁시킨 후 2N NaOH 수용액 (4.8 ml, 9.67 mmol)을 가하고 상온에서 1시간 동안 교반하였다. 반응액을 1N HCl 수용액을 이용하여 pH = 2로 조정한 뒤 EtOAc (20 ml x 2)로 추출하였다. 유기층을 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 흰색 고체 320 mg (70%)를 수득하였다.
단계2: tert-butyl ((1-(5-((2-chloro-3-(5-fluoropicolinamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate의 합성
5-fluoropicolinic acid (30 mg, 0.214 mmol)를 DCM (3 ml)에 현탁시킨 후 oxalyl chloride (0.05 ml, 0.56 mmol), 및 DMF (3 drops)를 가하고 상온에서 1 시간 동안 교반하였다. 반응액을 농축하여 얻은 노란색 오일을 DCM (1 ml)에 녹인 후 tert-butyl ((1-(5-((3-amino-2-chlorophenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (중간체 3, 50 mg, 0.11 mmol), 및 DIPEA (0.9 ml, 0.53 mmol)를 가하고 상온에서 1시간 동안 교반하였다. 반응액에 증류수 (10 ml)를 가한 뒤 DCM (10 ml x 2)로 추출하고 유기층을 무수황산나트륨으로 건조한 후 여과, 감압 농축하여 얻어진 잔사를 MPLC (0-100% EtOAc/Hexane)하여 노란색 고체 33 mg (53%)를 수득하였다.
단계 3: tert-butyl ((1-(5-((2-chloro-3-(5-(4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazin-1-yl)picolinamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate의 합성
tert-butyl ((1-(5-((2-chloro-3-(5-fluoropicolinamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (32 mg, 0.06 mmol), 및 (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)-2-(2-(piperazin-1-yl)ethoxy)benzyl)pyrrolidine-2-carboxamide (39 mg, 0.06 mmol)를 DMSO (0.2 ml)에 현탁시킨 후 DIPEA (0.03 ml, 0.17 mmol)를 가하고 75 ℃에서 16시간 동안 교반하였다. 반응액에 증류수 (10 ml)를 가한 뒤 EtOAc (10 ml x 2)로 추출하였다. 유기층을 brine (10 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-5% MeOH/DCM)하여 상아색 고체 36 mg (54%)을 수득하였다.
단계 4: N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-5-(4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazin-1-yl)picolinamide의 합성
tert-butyl ((1-(5-((2-chloro-3-(5-(4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazin-1-yl)picolinamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (33 mg, 0.03 mmol)를 DCM (2 ml)에 현탁시킨 후 4 M HCl in dioxane (0.3 ml)를 가하고 상온에서 1 시간 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (10 ml)을 가한 뒤 10% MeOH/DCM (10 ml x 2)로 추출하였다. 유기층을 brine (10 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (amine-silica, 0-5% MeOH/DCM)하여 상아색 고체 23 mg (77%)을 수득하였다.
실시예 82. N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-6-(4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazin-1-yl)nicotinamide
Figure PCTKR2022011797-appb-img-000107
methyl 5-fluoropicolinate 대신 methyl 6-fluoropicolinate를 사용하여 실시예 81의 합성법과 동일한 방법으로 실시예 82를 합성하였다.
실시예 83. N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-5-(4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazin-1-yl)pyrimidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000108
5-fluoropicolinic acid 대신 5-chloropyrimidine-2-carboxylic acid를 사용하여 실시예 81의 합성법과 동일한 방법으로 실시예 84를 합성하였다.
실시예 84. N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-2-(4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazin-1-yl)pyrimidine-5-carboxamide
Figure PCTKR2022011797-appb-img-000109
5-fluoropicolinic acid 대신 2-chloropyrimidine-5-carboxylic acid를 사용하여 실시예 81의 합성법과 동일한 방법으로 실시예 84를 합성하였다.
실시예 85. N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-6-(4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazin-1-yl)pyridazine-3-carboxamide
Figure PCTKR2022011797-appb-img-000110
methyl 5-fluoropicolinate 대신 methyl 6-chloropyridazine-3-carboxylate를 사용하여 실시예 81의 합성법과 동일한 방법으로 실시예 85를 합성하였다.
실시예 86. N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-2-(4-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)piperazin-1-yl)pyrimidine-4-carboxamide
Figure PCTKR2022011797-appb-img-000111
5-fluoropicolinic acid 대신 2-chloropyrimidine-4-carboxylic acid를 사용하여 실시예 81의 합성법과 동일한 방법으로 실시예 86을 합성하였다.
실시예 87. N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-5-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)-7-azaspiro[3.5]nonan-7-yl)pyrimidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000112
5-fluoropicolinic acid 대신 5-chloropyrimidine-2-carboxylic acid를 사용하고, (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)-2-(2-(piperazin-1-yl)ethoxy)benzyl)pyrrolidine-2-carboxamide 대신 (2S,4R)-N-(2-(7-azaspiro[3.5]nonan-2-yloxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide를 사용하여 실시예 81의 합성법과 동일한 방법으로 실시예 87을 합성하였다.
실시예 88. N-(3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)-6-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)-7-azaspiro[3.5]nonan-7-yl)pyridazine-3-carboxamide
Figure PCTKR2022011797-appb-img-000113
Methyl 5-fluoropicolinate 대신 methyl 6-chloropyridazine-3-carboxylate를 사용하고, (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)-2-(2-(piperazin-1-yl)ethoxy)benzyl)pyrrolidine-2-carboxamide 대신 (2S,4R)-N-(2-((7-azaspiro[3.5]nonan-2-yl)oxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide를 사용하여 실시예 81의 합성법과 동일한 방법으로 실시예 88을 합성하였다.
실시예 89. 2-(4-(4-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-4-oxobutyl)piperidin-1-yl)-N-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)pyrimidine-5-carboxamide
Figure PCTKR2022011797-appb-img-000114
Figure PCTKR2022011797-appb-img-000115
단계 1: tert-butyl (2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)carbamate의 합성
(2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (중간체 7, 50 mg, 0.09 mmol), 및 tert-butyl (2-bromoethyl)carbamate (122 mg, 0.56 mmol)을 DMF (0.2 ml)에 현탁시킨 후 potassium carbonate (39 mg, 0.28 mmol)을 가하고 70 ℃에서 16시간 동안 교반하였다. 반응액에 증류수 (10 ml)를 가한 뒤 EtOAc (5 ml x 2)로 추출하였다. 유기층을 brine (5 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-20% MeOH/DCM)하여 상아색 고체 73 mg (quant.)을 수득하였다.
단계 2: (2S,4R)-N-(2-(2-aminoethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide의 합성
tert-butyl (2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)carbamate (73 mg, 0.11 mmol)를 DCM (3 ml)에 현탁시킨 후 4 M HCl in dioxane (1 ml)를 가하고 상온에서 4 시간 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (5 ml)을 가하고 10% MeOH/DCM (5 ml x 2)로 추출하였다. 유기층을 brine (5 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (amine-silica, 0-5% MeOH/DCM)하여 흰색 고체 22 mg (35%)을 수득하였다.
단계 3: methyl 2-(4-(4-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-4-oxobutyl)piperidin-1-yl)pyrimidine-5-carboxylate의 합성
tert-butyl ((1-(5-((2-chloro-3-(3-(piperidin-4-yl)butanamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (100 mg, 0.16 mmol), methyl 2-chloropyrimidine-5-carboxylate (28 mg, 0.16 mmol), 및 DIPEA (0.08 ml, 0.48 mmol)를 DCM (0.6 ml)에 현탁시킨 후 상온에서 16시간 동안 교반하였다. 반응액에 증류수 (10 ml)를 가한 뒤 EtOAc (5 ml x 2)로 추출하였다. 유기층을 brine (5 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-100% EtOAc/Hexane)하여 상아색 고체 111 mg (91%)을 수득하였다.
단계 4: 2-(4-(4-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-4-oxobutyl)piperidin-1-yl)pyrimidine-5-carboxylic acid의 합성
Methyl 2-(4-(4-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-4-oxobutyl)piperidin-1-yl)pyrimidine-5-carboxylate (110 mg, 0.13 mmol), lithium hydroxide (20 mg, 0.53 mmol)를 water : THF = 1 : 1 (0.8 ml)에 현탁시킨 후 60 ℃에서 5시간 동안 교반하였다. 반응액을 1N HCl 수용액을 이용하여 pH = 2로 조정한 뒤 EtOAc (10 ml x 2)로 추출하고 유기층을 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 상아색 고체 92 mg (85%)를 수득하였다.
단계 5: tert-butyl ((1-(5-((2-chloro-3-(4-(1-(5-((2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)carbamoyl)pyrimidin-2-yl)piperidin-4-yl)butanamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate의 합성
2-(4-(4-((3-((5-(4-(((tert-butoxycarbonyl)amino)methyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-4-oxobutyl)piperidin-1-yl)pyrimidine-5-carboxylic acid (31 mg 0.04 mmol), (2S,4R)-N-(2-(2-aminoethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (20 mg, 0.03 mmol), EDCI (8 mg, 0.04 mmol), HOBt (6 mg, 0.04 mmol), 및 DIPEA (0.03 ml, 0.17 mmol)를 DMF (0.1 ml)에 현탁시킨 후 상온에서 16시간 동안 교반하였다. 반응액에 증류수 (10 ml)를 가한 뒤 EtOAc (5 ml x 2)로 추출하였다. 유기층을 brine (5 ml x 2)로 씻고 무수 황산 나트륨으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (10% MeOH/DCM)하여 상아색 고체 36 mg (68%)을 수득하였다.
단계 6: 2-(4-(4-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-4-oxobutyl)piperidin-1-yl)-N-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)pyrimidine-5-carboxamide의 합성
tert-butyl ((1-(5-((2-chloro-3-(4-(1-(5-((2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethyl)carbamoyl)pyrimidin-2-yl)piperidin-4-yl)butanamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (34.4 mg, 0.03 mmol)를 DCM (1 ml)에 현탁시킨 후 4 M HCl in dioxane (0.3 ml)를 가하고 상온에서 4 시간 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (5 ml)을 가하고 10% MeOH/DCM (5 ml x 2)로 추출하였다. 유기층을 brine (5 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (amine-silica, 0-5% MeOH/DCM)하여 상아색 고체 12 mg (39%)을 수득하였다.
실시예 90. (2S,4R)-N-(2-(2-((2-(4-(4-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-4-oxobutyl)piperidin-1-yl)pyrimidin-5-yl)oxy)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
Figure PCTKR2022011797-appb-img-000116
단계 1: tert-butyl ((1-(5-((2-chloro-3-(4-(1-(5-hydroxypyrimidin-2-yl)piperidin-4-yl)butanamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate의 합성
tert-butyl ((1-(5-((2-chloro-3-(3-(piperidin-4-yl)butanamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (65 mg, 0.11 mmol), 및 2-chloro-5-hydroxypyrimidine (21 mg, 0.16 mmol)을 DMF (0.3 ml)에 현탁시킨 후 potassium carbonate (34 mg, 0.24 mmol)을 가하고 마이크로웨이브에서 180 ℃로 16시간 동안 교반하였다. 반응액에 증류수 (10 ml)를 가한 뒤 EtOAc (5 ml x 2)로 추출하였다. 유기층을 brine (5 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-100% EtOAc/Hexane)하여 상아색 고체 20 mg (27%)을 수득하였다.
단계 2: tert-butyl ((1-(5-((2-chloro-3-(4-(1-(5-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethoxy)pyrimidin-2-yl)piperidin-4-yl)butanamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate의 합성
tert-butyl ((1-(5-((2-chloro-3-(4-(1-(5-hydroxypyrimidin-2-yl)piperidin-4-yl)butanamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (20 mg, 0.03 mmol), 및 (2S,4R)-N-(2-(2-chloroethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (중간체 10, 20 mg, 0.03 mmol)을 DMF (0.05 ml)에 현탁시킨 후 potassium carbonate (12 mg, 0.08 mmol), 및 potassium iodide (9 mg, 0.06 mmol)을 가하고 70 ℃에서 16시간 동안 교반하였다. 반응액에 증류수 (10 ml)를 가한 뒤 EtOAc (5 ml x 2)로 추출하였다. 유기층을 brine (5 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 MPLC (0-100% EtOAc/Hexane)하여 상아색 고체 35 mg (98%)을 수득하였다.
단계 3: (2S,4R)-N-(2-(2-((2-(4-(4-((3-((5-(4-(aminomethyl)-4-methylpiperidin-1-yl)pyrazin-2-yl)thio)-2-chlorophenyl)amino)-4-oxobutyl)piperidin-1-yl)pyrimidin-5-yl)oxy)ethoxy)-4-(4-methylthiazol-5-yl)benzyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide
tert-butyl ((1-(5-((2-chloro-3-(4-(1-(5-(2-(2-(((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)methyl)-5-(4-methylthiazol-5-yl)phenoxy)ethoxy)pyrimidin-2-yl)piperidin-4-yl)butanamido)phenyl)thio)pyrazin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate (33 mg, 0.03 mmol)를 DCM (1 ml)에 현탁시킨 후 4 M HCl in dioxane (0.3 ml)를 가하고 상온에서 4 시간 동안 교반하였다. 반응액에 NaHCO3 포화수용액 (5 ml)을 가하고 10% MeOH/DCM (5 ml x 2)로 추출하였다. 유기층을 brine (5 ml x 2)로 씻고 무수 황산 마그네슘으로 건조한 후 여과, 감압 농축하여 잔사를 얻었다. 얻어진 잔사를 PTLC (DCM : MeOH : NH4OH = 15 : 1 : 0.1)하여 상아색 고체 7.3 mg (24%)을 수득하였다.
상기 실시예 1-90에서 얻은 화합물들의 NMR 및 mass 측정 결과를 종합하여 하기 표 3에 나타내었다.
Figure PCTKR2022011797-appb-img-000117
Figure PCTKR2022011797-appb-img-000118
Figure PCTKR2022011797-appb-img-000119
Figure PCTKR2022011797-appb-img-000120
Figure PCTKR2022011797-appb-img-000121
Figure PCTKR2022011797-appb-img-000122
Figure PCTKR2022011797-appb-img-000123
Figure PCTKR2022011797-appb-img-000124
Figure PCTKR2022011797-appb-img-000125
Figure PCTKR2022011797-appb-img-000126
Figure PCTKR2022011797-appb-img-000127
본 발명 화합물들의 평가
실험예 1: KYSE-520 세포에서 SHP2 단백질 분해능 측정
합성된 실시예 화합물을 KYSE-520 세포에 처리하고 세포내에 존재하는 SHP2 단백질의 양을 western blotting 검출법을 사용하여 측정하였다. KYSE-520 세포를 사용하여 진행된 실험의 protocol은 다음과 같다.
[배양] KYSE-520 세포를 5% FBS (Hyclone, SV30207.02) 및 1% Penicillin-streptomycin (Welgene, LS 202-02)를 포함하는 RPMI1640 MEDIUM (Hyclone, SH30027.01)에 resuspend 하여 2.5 x 105/well로 6-well plate에 2 mL씩 seeding 후 2일 동안 배양한다.
[화합물 처리] 10 mM stock을 DMSO로 1/10씩 serial dilution하여 (3 mL + 27 mL DMSO) 최종 농도 0.1, 1, 10 mM 또는 0.001, 0.01, 0.1 mM로 화합물을 세포에 처리하고 24 시간 후 harvest한다. 음성대조군에는 media에 1/10로 희석한 DMSO를 넣어준다 (3 mL DMSO + 27 mL media).
[세포 용해 및 단백질 정량] Iysis buffer는 다음과 같이 준비한다. RIPA buffer (Biosesang, RC2038-050-00) + 0.5 mM PMSF (SIGMA, P7626) + 1x Protease/Phosphatase Inhibitor (Cell signaling, 5872S)를 well 당 60 mL씩 넣고 30분간 얼음위에 둔다 (0, 30분에 각각 vortexing). 이후 sonication 과정을 (10초 pulse, 30초 rest, 5 cycles, 70% amplification) 진행하고 centrifuge (15000 g, 15 min, 4℃)한 후 상층액만 취하여 새 microtube로 옮긴다. 96-well plate에 RIPA buffer로 샘플 1/4 희석해서 넣고, BCA Protein Assay Kit (iNtRON, 21071)의 A:B = 50:1로 섞어 200 mL씩 넣어 37℃에서 30분 동안 두고 10분간 식혀준다. 이후 BioTek 사의 SYNERGY H1 microplate reader로 562 nm에서 absorbance를 측정한다. 측정값으로 단백질을 정량하여 샘플을 만든 후 70℃에서 10분간 boiling한다. 이때 사용되는 Sample buffer는 사용할 gel에 맞추어 NuPAGE 혹은 Bolt 4x sample buffer (Invitrogen)와 각각의 10 x sample reducing agent를 섞어서 사용하며 단백질 희석은 RIPA buffer를 이용한다.
[Western blotting 검출법] NuPAGE 혹은 Bolt Bis-tris 4-12% gradient gel에 같은 양의 샘플을 loading 하여 running한다 (200 V, 35분). Trans-blot Turbo (BIO-RAD)로 0.2 mm NC membrane에 transfer 한다 (1.3 A constant, 25 V limit, 15분). Skim milk 혹은 Intercept Blocking Buffer (LI-COR, 927-60001):0.1% TBST = 1:1 로 1시간 동안 상온에서 blocking 한다. Anti SHPTP2 Mouse (1:500 in 5% Skim milk/TBST, size: 70 kDa, Santacruz sc-7384)은 4℃에서 overnight incubation 하고, Anti GAPDH Rabbit (1:2,000 in 5% BSA/0.2% TBST, size: 37 kDa, CST #2118)은 상온에서 1시간 30분 혹은 3시간 동안 incubation 한다. 0.2% TBST washing buffer로 각 5분씩 3회 washing 해준다. 2차 antibody인 Anti-Rabbit IgG (1:5,000 in 5% Skim milk/0.2% TBST, CST), 및 IRDye® 680RD Goat anti-Rabbit IgG Secondary Antibody (1:10,000 in 5% BSA/0.2% TBST, LI-COR 926-68071)를 상온의 rocker 위에서 45분간 incubation 한다. 0.2% TBST washing buffer로 각 5분씩 5회 washing 해주고 LI-COR사의 Odyssey로 detection 한다. 이때 SHPTP2은 SuperSignal West Pico PLUS Chemiluminescent Substrate를 이용해 detection하고 house-keeping gene들도 함께 detection한다.
실험예 2: MV-4-11 세포에서 SHP2 단백질 분해능 측정
합성된 실시예 화합물을 MV-4-11 세포에 처리하고 세포내에 존재하는 SHP2 단백질의 양을 western blotting 검출법을 사용하여 측정하였다. MV-4-11 세포를 사용하여 진행된 실험의 protocol은 다음과 같다.
[배양] MV-4-11 세포를 IMDM MEDIUM (Hyclone, SH30228.01, 5% FBS (Hyclone, SV30207.02), 1% Penicillin-streptomycin (Welgene, LS 202-02)) media에 resuspend 하여 1 x 106/mL로 12-well plate에 1 mL씩 seeding 한다.
[화합물 처리] 12-well plate에 다음과 같이 처리한다. 10 mM stock을 DMSO 로 1/100씩 serial dilution하여 (3 mL + 297 mL DMSO) 최종 농도 0.01, 0.1, 1 mM 또는 0.001, 0.01, 0.1 mM로 화합물을 세포에 처리하고 24 시간 후 harvest 한다. 음성대조군에는 media에 1/100으로 희석한 DMSO를 넣어준다 (3 mL DMSO + 297 mL media).
[Harvest] 각 well을 1 mL pipette으로 pipetting 하여 1.5 mL microtube에 모아 centrifuge (500 g, 5 min, 4℃)한다. 상층액을 제거하고 PBS washing하여 다시 centrifuge (500 g, 5 min, 4℃)한다. 이후 상층액을 제거하여 pellet을 준비한다.
[Cell lysis 및 Sample 준비] Iysis buffer는 다음과 같이 준비한다. RIPA buffer (Biosesang, RC2038-050-00) + 0.5 mM PMSF (SIGMA, P7626) + 1x Protease/Phosphatase Inhibitor (Cell signaling, 5872S)를 well 당 50 mL씩 넣고 30분간 얼음위에 방치한다 (0, 30분에 각각 vortexing). 이후 sonication 과정을 (10초 pulse, 30초 rest, 5 cycles, 70% amplification) 진행하고 centrifuge (15000 g, 15 min, 4℃)한 후 상층액만 취하여 microtube로 옮긴다. 96-well plate에 RIPA buffer로 샘플 1/4 희석해서 넣고 BCA Protein Assay Kit (iNtRON, 21071)의 A:B = 50:1로 섞어 200 mL씩 넣어 37℃에서 30분 동안 두고 10분간 방치한다. 이후 BioTek 사의 SYNERGY H1 microplate reader로 562 nm에서 absorbance를 측정한다. 측정값으로 단백질을 정량하여 샘플을 만든 후 70℃에서 10분간 boiling 한다. 이때 사용되는 Sample buffer는 사용할 gel에 맞추어 NuPAGE 혹은 Bolt 4x sample buffer (Invitrogen)와 각각의 10x sample reducing agent를 섞어서 사용하며 단백질 희석은 RIPA buffer를 이용한다.
[Western blotting 검출법] NuPAGE 혹은 Bolt Bis-tris 4-12% gradient gel에 같은 양의 샘플을 loading하여 running 한다 (200 V, 35분). Trans-blot Turbo (BIO-RAD)로 0.2 mm NC membrane에 transfer 한다 (1.3 A constant, 25 V limit, 15분). Skim milk 혹은 Intercept Blocking Buffer (LI-COR, 927-60001):0.1% TBST = 1:1 로 1시간 동안 상온에서 blocking 한다. Anti-SHPTP2 Mouse (1:500 in 2% BSA/0.2% TBST, size: 70 kDa, Santacruz sc-7384)은 4℃에서 overnight incubation 하고, Anti-GAPDH Rabbit (1:2,000 in 2% BSA/0.2% TBST, size: 37 kDa, CST #2118)은 상온에서 1시간 30분 혹은 3시간 동안 incubation한다. 0.2% TBST washing buffer로 각 5분씩 3회 washing해주고 2차 antibody인 Anti-Rabbit IgG (1:5,000 in 2% BSA/0.2% TBST, CST), IRDye® 680RD Goat anti-Rabbit IgG Secondary Antibody (1:10,000 in 2% BSA/0.2% TBST, LI-COR 926-68071)를 넣어준 후 상온의 rocker 위에서 45분간 incubation한다. 0.2% TBST washing buffer로 각 5분씩 5회 washing 해주고 LI-COR사의 Odyssey로 detection해준다. 이때 SHPTP2은 SuperSignal West Pico PLUS Chemiluminescent Substrate를 이용해 detection하고 house-keeping gene들도 함께 detection한다.
본 발명의 화합물들 SHP2 단백질 분해능 평가 결과를 하기 표 4에 종합하여 나타내었다. 하기 표 4의 결과에서 화합물 1-7, 13-19, 및 21-26은 KYSE-520 세포에서의 결과이며, 나머지들은 MV-4-11 세포에서의 결과이다.
Figure PCTKR2022011797-appb-img-000128
Figure PCTKR2022011797-appb-img-000129
Figure PCTKR2022011797-appb-img-000130
상기 표 4에 나타나는 바와 같이, 본 발명에 따른 화합물들은 SHP2 단백질 분해에 있어 우수한 활성을 나타내었다.
특히, 본 발명의 화합물 20, 29-45, 50, 52-58, 60-64, 66-68, 76-80, 및 83-85는 유사한 구조를 가진 다른 화합물들보다 우수한 SHP2 단백질 분해 활성을 나타내었으며, 본 발명의 화합물 29, 31, 33, 36-38, 43, 50, 52-58, 61-63, 66, 68, 76-80, 및 83는 다른 화합물들보다 월등히 우수한 SHP2 단백질 분해 활성을 나타내었다.
한편, 본 발명의 화학식 1에서
Figure PCTKR2022011797-appb-img-000131
부분에 포함된 malonamide 구조의 두개의 carbonyl group 사이에 존재하는 탄소에 두개의 치환기가 있거나 사이클로프로필로 치환된 경우(예를 들어, 화합물 46, 47, 48, 49, 59)에는 분해 활성이 다소 떨어졌다. 따라서, 본 발명의 더욱 더 바람직한 양태에 있어, 본 발명의 화학식 1에서
Figure PCTKR2022011797-appb-img-000132
부분에 포함된 malonamide 구조의 두개의 carbonyl group 사이에 존재하는 탄소에 두개의 치환기가 있거나 사이클로프로필로 치환된 화합물들은 제외된다.

Claims (13)

  1. 하기 화학식 1의 화합물 또는 이의 약학적으로 허용 가능한 염.
    [화학식 1]
    Figure PCTKR2022011797-appb-img-000133
    상기 화학식 1에서,
    R1a, R1b, R1c, 및 R1d는 서로 독립적으로 H 또는 C1-6alkyl이고,
    R2는 H, C1-6alkyl, 또는 -NH2이고,
    R3는 H, C1-6alkyl, 또는 C1-6hydroxyalkyl이고,
    R4는 H, C1-6alkyl, halogen, 또는 C1-6haloalkyl이고,
    R5는 H, C1-6alkyl, C1-6alkynyl, halogen, -CN, 또는 heteroaryl이며, 여기에서 헤테로아릴은 임의로 고리 내 하나 이상의 수소가 C1-6alkyl, halogen, 또는 C1-3haloalkyl로 치환되며,
    R6는 -NHC(O)R7 또는 heteroaryl이고, 여기에서 헤테로아릴은 임의로 고리 내 하나 이상의 수소가 C1-6alkyl, halogen, 또는 C1-3haloalkyl로 치환되며,
    R7는 C1-6alkyl, C1-6haloalkyl, C3-10사이클로알킬, 또는 헤테로사이클이고, 여기에서 사이클로알킬 및 헤테로사이클은 서로 독립적이면서 임의로 고리 내 하나 이상의 수소가 C1-6alkyl, halogen, 또는 -CN로 치환되며,
    X는 CH 또는 N이고,
    n1 및 n2는 서로 독립적으로 0, 1, 또는 2이고,
    m은 0 또는 1이며,
    L은 하기 화학식 2이며,
    [화학식 2]
    Figure PCTKR2022011797-appb-img-000134
    화학식 2에서,
    A1, A2, 및 A3은 서로 독립적으로 직접 결합, -O-, -N(R9)- -C(O)-, -C(O)N(R9)-, 또는 -N(R9)C(O)-이고,
    B1 및 B2는 서로 독립적으로 직접 결합, C3-10사이클로알킬, 헤테로사이클, 아릴, 또는 헤테로아릴이며, 여기에서 사이클로알킬, 헤테로사이클, 아릴, 또는 헤테로아릴은 임의로 고리 내 하나 이상의 수소가 C1-6alkyl, halogen, C1-6haloalkyl, -OH, =O, 또는 -CO2R9로 치환되며,
    R8 및 R8'는 서로 독립적으로 H 또는 C1-6alkyl이거나, R8 및 R8'는 서로 연결되어 C3-10사이클로알킬을 형성하고,
    R9는 수소 또는 C1-6alkyl이고,
    q1, q2, q3, q4, 및 q5는 서로 독립적으로 0 내지 10의 정수임.
  2. 제1항에 있어서, 상기 화학식 1에서,
    R1a, R1b, R1c, 및 R1d는 서로 독립적으로 H 또는 C1-6alkyl이고,
    R2는 H, C1-6alkyl, 또는 -NH2이고,
    R3는 H, C1-6alkyl, 또는 C1-6hydroxyalkyl이고,
    R4는 H, C1-6alkyl, 또는 halogen이고,
    R5는 heteroaryl이며, 여기에서 헤테로아릴은 임의로 고리 내 하나 이상의 수소가 C1-6alkyl 또는 halogen으로 치환되며,
    R6는 -NHC(O)R7이고,
    R7는 C1-6alkyl 또는 C3-10사이클로알킬이고, 여기에서 사이클로알킬은 임의로 고리 내 하나 이상의 수소가 C1-6alkyl, halogen, 또는 -CN로 치환되며,
    X는 CH 또는 N이고,
    n1 및 n2는 서로 독립적으로 0 또는 1이고,
    m은 0 또는 1이며,
    L은 하기 화학식 2이며,
    [화학식 2]
    Figure PCTKR2022011797-appb-img-000135
    화학식 2에서,
    A1, A2, 및 A3은 서로 독립적으로 직접 결합, -O-, -N(R9)- -C(O)-, -C(O)N(R9)-, 또는 -N(R9)C(O)-이고,
    B1 및 B2는 서로 독립적으로 직접 결합, 헤테로사이클, 또는 헤테로아릴이며, 여기에서 헤테로사이클 또는 헤테로아릴은 임의로 고리 내 하나 이상의 수소가 C1-6alkyl, halogen, -OH, =O, 또는 -CO2R9로 치환되며,
    R8 및 R8'는 서로 독립적으로 H 또는 C1-6alkyl이거나, R8 및 R8'는 서로 연결되어 C3-10사이클로알킬을 형성하고,
    R9는 수소 또는 C1-6alkyl이고,
    q1, q2, q3, q4, 및 q5는 서로 독립적으로 0 내지 10의 정수인,
    화학식 1의 화합물 또는 이의 약학적으로 허용 가능한 염.
  3. 제1항에 있어서, 상기 화학식 1에서,
    R1a, R1b, R1c, 및 R1d는 서로 독립적으로 H 또는 C1-6alkyl이고,
    R2는 H 또는 -NH2이고,
    R3는 H이고,
    R4는 H 또는 halogen이고,
    R5는 thiazole이며, 여기에서 thiazole은 임의로 고리 내 하나 이상의 수소가 C1-6alkyl로 치환되며,
    R6는 -NHC(O)R7이고,
    R7는 C1-6alkyl 또는 사이클로프로필이고, 여기에서 사이클로프로필은 임의로 고리 내 하나 이상의 수소가 halogen 또는 -CN으로 치환되며,
    X는 CH이고,
    n1 및 n2는 서로 독립적으로 0 또는 1이고,
    m은 0 또는 1이며,
    L은 하기 화학식 2이며,
    [화학식 2]
    Figure PCTKR2022011797-appb-img-000136
    화학식 2에서,
    A1, A2, 및 A3은 서로 독립적으로 직접 결합, -O-, -N(R9)- -C(O)-, -C(O)N(R9)-, 또는 -N(R9)C(O)-이고,
    B1 및 B2는 서로 독립적으로 직접 결합, 아제티딘, 피롤리딘, 피페리딘, 피페라진, azaspiro[3.5]nonane, diazabicyclo[3.2.1]octane, 피리딘, 피리미딘, 피리다진, 또는 피라진이며, 여기에서 아제티딘, 피롤리딘, 피페리딘, 피페라진, azaspiro[3.5]nonane, diazabicyclo[3.2.1]octane, 피리딘, 피리미딘, 피리다진, 또는 피라진은 임의로 고리 내 하나 이상의 수소가 C1-6alkyl, -OH, =O, 또는 -CO2R9로 치환되며,
    B1 및 B2 중 어느 하나 이상은 아제티딘, 피롤리딘, 피페리딘, 피페라진, azaspiro[3.5]nonane, diazabicyclo[3.2.1]octane, 피리딘, 피리미딘, 피리다진, 또는 피라진이고,
    R8 및 R8'는 서로 독립적으로 H 또는 C1-6alkyl이거나, R8 및 R8'는 서로 연결되어 C3-10사이클로알킬을 형성하고,
    R9는 수소 또는 C1-6alkyl이고,
    q1, q2, q3, q4, 및 q5는 서로 독립적으로 0 내지 10의 정수인,
    화학식 1의 화합물 또는 이의 약학적으로 허용 가능한 염.
  4. 제1항에 있어서, 상기 L은 하기들 중 어느 하나인, 화학식 1의 화합물 또는 이의 약학적으로 허용 가능한 염.
    Figure PCTKR2022011797-appb-img-000137
    Figure PCTKR2022011797-appb-img-000138
  5. 제1항에 있어서, 상기 화합물은 하기 화합물들 중 어느 하나인 화합물 또는 이의 약학적으로 허용 가능한 염.
    Figure PCTKR2022011797-appb-img-000139
    Figure PCTKR2022011797-appb-img-000140
    Figure PCTKR2022011797-appb-img-000141
    Figure PCTKR2022011797-appb-img-000142
    Figure PCTKR2022011797-appb-img-000143
    Figure PCTKR2022011797-appb-img-000144
  6. 제5항에 있어서, 상기 화합물은 하기 화합물들 중 어느 하나인 화합물 또는 이의 약학적으로 허용 가능한 염.
    Figure PCTKR2022011797-appb-img-000145
    Figure PCTKR2022011797-appb-img-000146
    Figure PCTKR2022011797-appb-img-000147
  7. 제1항 내지 제6항 중 어느 한 항의 화합물 또는 이의 약학적으로 허용 가능한 염, 및 약학적으로 허용 가능한 담체를 포함하는 조성물.
  8. 유효 성분으로 제1항 내지 제4항 중 어느 한 항의 화합물 또는 이의 약학적으로 허용 가능한 염을 포함하는, Src 상동성 2 도메인-함유 포스파타제(Src homology 2 domain-containing phosphatase, SHP2)의 억제 또는 분해를 위한 약학 조성물.
  9. 제8항에 있어서, 상기 약학 조성물은 암의 치료 또는 예방용인, 약학 조성물.
  10. 제9항에 있어서, 상기 암은 백혈병, 림프종, 폐암, 두경부암, 식도암, 위암, 대장암, 췌장암, 간암, 유방암, 난소암, 자궁경부암, 방광암, 흑색종, 신경모세포종, 신경교종, 또는 육종인, 약학 조성물.
  11. 제1항 내지 제6항 중 어느 한 항의 화합물 또는 이의 약학적으로 허용 가능한 염의 치료적으로 유효한 양을 이를 필요로 하는 개체에게 투여하는 단계를 포함하는, Src 상동성 2 도메인-함유 포스파타제 관련 질환의 치료 또는 예방 방법.
  12. 제11항에 있어서, 상기 Src 상동성 2 도메인-함유 포스파타제 관련 질환은 암인 치료 또는 예방 방법.
  13. 제12항에 있어서, 상기 암은 백혈병, 림프종, 폐암, 두경부암, 식도암, 위암, 대장암, 췌장암, 간암, 유방암, 난소암, 자궁경부암, 방광암, 흑색종, 신경모세포종, 신경교종, 또는 육종인, 치료 또는 예방 방법.
PCT/KR2022/011797 2021-08-09 2022-08-08 Shp2 단백질 분해활성을 갖는 화합물 및 이들의 의약 용도 Ceased WO2023018155A1 (ko)

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