Novel compound having SOS1 inhibitory activity

Abstract

Provided are a novel compound of chemical formula I and uses thereof for the prevention or treatment of SOS1-related diseases. A novel compound according to an aspect of the present invention is useful for preventing or treating SOS1-mediated diseases such as cancer and RASopathy by inhibiting interaction between SOS1 and a RAS family protein or between SOS1 and RAC1. The compounds of the present invention exhibit optimal DMPK properties and favorable PK exposure, and provide excellent effects due to a reduced risk of drug–drug interactions. In addition, the compound of the present invention exhibits improved tumor inhibitory activity when administered in combination with other anticancer agents, for example, KRAS inhibitors, and thus can be advantageously used for the prevention and treatment of cancer.

Description

Novel compounds with SOS1 inhibitory activity

[0001] The present invention relates to a novel compound having SOS1 inhibitory activity, a solvate, stereoisomer or pharmaceutically acceptable salt thereof, a pharmaceutical composition for preventing or treating a disease comprising the same as an active ingredient, and a pharmaceutical use thereof.

[0002] Mutations in the RAS gene are a major oncogene with a high incidence in human cancer, occurring in 20–30% of human cancers, and are particularly prevalent in lung, colon, rectal, and pancreatic cancers. RAS-family proteins include KRAS, NRAS, and HRAS.

[0003] RAS proteins are small GTPases that exist in cells in either GTP- or GDP-bound states, functioning as molecular switches that cycle between an active GTP-bound state and an inactive GDP-bound state. Mutations in the RAS gene reduce the ability of the GTPase RAS to hydrolyze GTP, thereby maintaining this molecular switch in a constitutively active GTP-bound form, thereby inducing oncogenic downstream signaling pathways (e.g., the Raf-MEK-ERK pathway or the PI3K-PDK1-Akt pathway).

[0004] Meanwhile, binding of GTPase-activating proteins (GAPs), such as NF1, accelerates the weak intrinsic GTPase activity of RAS proteins, downregulating active RAS and reverting it to an inactive form. Conversely, binding of guanine nucleotide exchange factors (GEFs), such as SOS1, promotes GDP release from RAS proteins and increases the GTP-bound active state.

[0005] RAS has long been considered an undruggable target, but recent advances in structure-based drug design have led to rapid progress in drug development. Sotorasib and adagrasib, which specifically bind to the KRAS G12C mutation, have received FDA approval, and KRAS inhibitors targeting other mutations, such as G12D and G12V, as well as pan-KRAS inhibitors, are also in development. Furthermore, to overcome signaling pathway reactivation and resistance that occur with single-target inhibition, combination therapies of KRAS inhibitors with downstream RAS-RAF-MEK pathway inhibitors or inhibitors acting on upstream regulators of the RAS pathway, such as MEK inhibitors and SOS1 inhibitors, are also actively under development.

[0006] Son of Sevenless 1 (SOS1) is a guanine nucleotide exchange factor (GEF) that promotes GDP release from RAS family proteins, allowing GTP binding, thereby regulating RAS family protein signaling. Son of Sevenless (SOS) exists in two isoforms, SOS1 and SOS2, and only SOS1 is phosphorylated by ERK. Growth factor-induced phosphorylation of SOS1 is largely mediated by ERK, which phosphorylates at least four serine residues in the C-terminal domain of SOS1. This suggests that SOS1 plays a crucial role in the negative feedback regulation of the KRAS pathway. The SOS1 protein consists of 1333 amino acids (150 kDa). SOS1 is a multidomain protein with a Dbl homology domain (DH) followed by two tandem N-terminal histone domains (HD), a pleckstrin homology domain (PH), a helical linker (HL), a RAS exchange motif (REM), a CDC25 homology domain, and a C-terminal proline-rich domain (PR). SOS1 has two binding sites for RAS family proteins: a catalytic site that binds GDP-bound RAS family proteins to catalyze the exchange of guanine nucleotides, and an allosteric site that upregulates the catalytic site activity of SOS1 by binding GTP-bound RAS family proteins (J. Med. Chem. 2021, 64, 10, 6569-6580). Selective pharmacological inhibition of SOS1 binding to the catalytic site of RAS family proteins is expected to prevent SOS1-mediated activation of RAS family proteins in the GTP-bound form.

[0007] Therefore, novel SOS1 inhibitor compounds are being developed that bind to the SOS1 catalytic site and prevent binding to and activation of RAS family proteins, as SOS1 inhibitor compounds are expected to inhibit signaling (e.g., ERK phosphorylation) downstream of RAS-family proteins.

[0008] SOS1 has been reported to be crucially involved in mutant KRAS activation and oncogenic signaling in cancer (Current Opinion in Chemical Biology, 2021, 62: 109-118). Depletion of SOS1 levels decreased the survival of tumor cells harboring KRAS mutations, but no such effect was observed in KRAS wild-type cell lines. The SOS1 depletion effect was observed in SOS1 cells with a damaged catalytic site. F929A It cannot be rescued by SOS1 mutants (SOS1L687E / R688A) that are defective in GTP-KRAS binding at the catalytic or allosteric site, suggesting that targeting the catalytic or allosteric site of SOS1 may be a viable option for the treatment of KRAS mutant cancers.

[0009] Furthermore, SOS1 is crucial for the activation of RAS family protein signaling in cancer through mechanisms other than mutations in RAS family proteins. SOS1 interacts with the adaptor protein Grb2 to form the SOS1 / Grb2 complex. This complex binds to activated / phosphorylated receptor tyrosine kinases (e.g., EGFR, ErbB2, ErbB3, ErbB4, PDGFR-A / B, FGFR1 / 2 / 3, IGF1R, INSR, ALK, ROS, TrkA, TrkB, TrkC, RET, c-MET, VEGFR1 / 2 / 3, AXL). SOS1 has also been reported to localize to other phosphorylated cell surface receptors, such as the T-cell receptor (TCR), B-cell receptor (BCR), and monocyte colony-stimulating factor receptor, resulting in the activation of RAS family proteins.

[0010] Furthermore, SOS1 is a GEF for the activation of the GTPase RAC1 (Ras-related C3 botulinum toxin substrate 1). RAC1, like other RAS-family proteins, is known to be involved in the pathogenesis of various cancers and other diseases.

[0011] Currently, BI-3406, BI-1701963, and MRTX0902 are under development as SOS1 inhibitors, and the present inventors are also developing novel compounds with SOS1 inhibitory activity (WO 2023 / 022497 A1). SOS1 inhibitors are primarily being developed as combination drugs with other anticancer agents, such as KRAS inhibitors and EGFR inhibitors. Therefore, reducing the risk of drug interactions is a critical challenge in the development of SOS1 inhibitors.

[0012] Therefore, research is ongoing in the art to develop an SOS1 inhibitor that not only has an excellent inhibitory effect on the interaction between SOS1 and RAS family proteins and / or RAC1, but also has minimal drug interactions and excellent pharmacodynamic characteristics.

[0013] The present inventors developed a compound having SOS1 inhibitory activity (WO 2023 / 022497 A1) and, while continuing research on the compound, completed the present invention by newly developing an SOS1 inhibitor having excellent SOS1 inhibitory activity, a reduced risk of drug interaction, and excellent pharmacodynamic characteristics.

[0014] An object of the present invention is to provide a compound of formula I, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof.

[0015] Another object of the present invention is to provide a pharmaceutical composition comprising a compound of formula I, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof.

[0016] Another object of the present invention is to provide a method for preventing or treating an SOS1-mediated disease by administering a compound of formula I, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof.

[0017] Another object of the present invention is to provide a use of a compound of formula I, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the prevention or treatment of SOS1-mediated diseases.

[0018] Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer, comprising a compound of formula I, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof; and an anticancer agent as active ingredients.

[0019] Each description and embodiment disclosed in this application may also be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in this application fall within the scope of this application. Furthermore, the scope of this application is not limited by the specific descriptions described below.

[0020]

[0021] One aspect of the present invention provides a compound represented by the following chemical formula I, or a solvate, stereoisomer or pharmaceutically acceptable salt thereof.

[0022] [Chemical Formula I]

[0023]

[0024] In the above chemical formula I,

[0025] Z 1 is N or CH,

[0026] A 1 is CF3, CH3-CF2- or CH2F-CF2-,

[0027] A 2 is halogen; or C1-C6 alkyl optionally substituted with one or more deuterium,

[0028] R 1 is H; or C1-C3 alkyl optionally substituted with halogen,

[0029] R 2 is a straight or branched C1-C6 alkyl optionally substituted with one or more deuterium or halogen; a C3-C7 cycloalkyl optionally substituted with one or more deuterium, halogen or C1-C3 alkyl; or a 3- to 7-membered heterocyclyl comprising a heteroatom or heteroatom group selected from N, O, S and SO2.

[0030]

[0031] In the above chemical formula I, Z 1 is N or CH. In one specific example, Z 1 may be CH, in which case the compound of formula I may be represented by the following formula IA:

[0032] [Chemical Formula IA]

[0033]

[0034] (In the above chemical formula IA, A 1 , A 2 , R 1 and R 2 is as described below for chemical formula I.)

[0035] In the above chemical formula I, A 1 is CF3, CH3-CF2- or CH2F-CF2-.

[0036] In the above chemical formula I, A 2 is halogen; or C1-C6 alkyl optionally substituted with one or more deuterium. In one specific embodiment, A 2 may be F, Cl, C1-C3 alkyl optionally substituted with one or more deuterium. For example, A 2 may be, but is not limited to, F, CH3 or CD3.

[0037] In the above chemical formula I, A 1 If this is CF3, A 2may be a halogen, a C1-C6 alkyl, or a C1-C6 alkyl substituted with deuterium. In one specific example, A 1 If this is CF3, A 2 is not F. In one specific example, A 1 If this is CF3, A 2 may be C1-C3 alkyl or C1-C3 alkyl substituted with deuterium.

[0038] In one specific example, can be selected from the following structures:

[0039] , and .

[0040] In the above chemical formula I, R 1 is H; or C1-C3 alkyl optionally substituted with halogen. In one specific embodiment, R 1 may be H, C1-C3 alkyl, or C1-C3 alkyl substituted with one or more F. For example, R 1 Silver H, CH 3, C2H 5, CF 3, CHF 2, or CH2F, but is not limited thereto. For example, R 1 Silver H, CH 3, Or it could be CF3.

[0041] In one specific example of the above chemical formula I, Z 1 If this is CH, R 1 can be H. Alternatively, Z 1 If this is N, then R 1 may be H, C1-C3 alkyl or C1-C3 haloalkyl. Here, C1-C3 haloalkyl may include, but is not limited to, C1-C3 alkyl substituted with one or more F.

[0042] In the above chemical formula I, R 2is a straight or branched C1-C6 alkyl optionally substituted with one or more deuterium or halogen; a C3-C7 cycloalkyl optionally substituted with one or more deuterium, halogen or C1-C3 alkyl; or a 3- to 7-membered heterocyclyl comprising a heteroatom or heteroatom group selected from N, O, S and SO2. In one specific embodiment, R 2 may be a straight or branched chain C1-C6 alkyl optionally substituted with one or more deuterium atoms. In one specific embodiment, R 2 is C3-C7cycloalkyl optionally substituted with one or more halogens or C1-C3alkyl; or may be a 3- to 7-membered heterocyclyl comprising a heteroatom or heteroatom group selected from N, O, S, and SO2. In one specific embodiment, R 2 may be C3-C7cycloalkyl optionally substituted with one or more halogens or C1-C3alkyl. For example, R 2 may be C3-C5 cycloalkyl optionally substituted with one or more halogens or C1-C3 alkyl. For example, R 2 may be a straight or branched chain C1-C3 alkyl (e.g., methyl, ethyl, isopropyl or CD3) optionally substituted with one or more deuterium atoms. For example, R 2 may be C3-C5cycloalkyl (e.g., cyclopropyl or cyclobutyl) optionally substituted with one or more halogens (e.g., F) or methyl. Alternatively, R 2 It may be a heterocyclyl selected from oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, 1,1-dioxidothietanyl and 1,1-dioxidotetrahydrothiophenyl. In this case, the heterocyclyl group may be optionally substituted with deuterium, halogen or C1-C3 alkyl.

[0043] For example, in chemical formula I, R 2 is CH3, CD3, -CH2CH3, -CH(CH3)2, cyclopropyl, cyclobutyl, or It may be, but is not limited to, R. For example, 2 may be CH3, CD3, or cyclopropyl optionally substituted with one or more F. For example, R 2 Is , , or It can be. In particular, R 2 When R is substituted with deuterium, methyl, cyclopropyl, or cyclopropyl substituted with one or more halogens (e.g., F), it may be advantageous in terms of reducing drug interactions and improving pharmacokinetic properties. Preferably, R 2 is CD3, cyclopropyl, , or It could be.

[0044] In one specific example, A of the above chemical formula I 2 is F or Cl, and Z 1 This is N, and R 1 If this is H, then R 2 is not CH3.

[0045] In one specific embodiment, A in formula I 2 , R 2 Or all of these can be C1-C6 alkyl substituted with deuterium. For example, A 2 , R 2 Alternatively, all of these may be C1-C3 alkyl substituted with deuterium, e.g., CD3.

[0046] In some embodiments, the compound of Formula I herein can be a compound represented by Formula IB:

[0047] [Chemical Formula IB]

[0048]

[0049] In the above chemical formula IB,

[0050] Z 1 is N or CH,

[0051] A 2is C1-C6 alkyl optionally substituted with one or more deuterium atoms,

[0052] R 2 is a straight or branched C1-C6 alkyl optionally substituted with one or more deuterium or halogen; a C3-C7 cycloalkyl optionally substituted with one or more deuterium, halogen or C1-C3 alkyl; or a 3- to 7-membered heterocyclyl comprising a heteroatom or heteroatom group selected from N, O, S and SO2.

[0053] In the above chemical formula IB, Z 1 is N or CH.

[0054] In the above chemical formula IB, A 2 is C1-C6 alkyl or C1-C6 alkyl substituted with one or more deuterium. In one specific example, A 2 may be C1-C3 alkyl or C1-C3 alkyl substituted with one or more deuteriums. For example, A 2 may be, but is not limited to, CH3 or CD3.

[0055] In the above chemical formula IB, R 2 is a straight or branched C1-C6 alkyl optionally substituted with one or more deuterium or halogen; a C3-C7 cycloalkyl optionally substituted with one or more deuterium, halogen or C1-C3 alkyl; or a 3- to 7-membered heterocyclyl comprising a heteroatom or heteroatom group selected from N, O, S and SO2. In one specific embodiment, R 2 may be a straight or branched chain C1-C3 alkyl optionally substituted with one or more deuterium atoms. For example, R 2 may be, but is not limited to, CH3 or CD3. In one specific embodiment, R 2 may be C3-C5 cycloalkyl optionally substituted with one or more halogens or C1-C3 alkyl. For example, R 2is cyclopropyl or cyclobutyl, which may be optionally substituted with one or more F or methyl. In one embodiment, R 2 may be a heterocyclyl selected from oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, 1,1-dioxidothietanyl and 1,1-dioxidotetrahydrothiophenyl. For example, R 2 can be oxetanyl, tetrahydrofuranyl, 1,1-deoxydothietanyl. For example, R 2 Is It could be.

[0056] In one specific example of the above chemical formula IB, Z 1 is N or CH, and A 2 and R 2 are each independently C1-C6 alkyl optionally substituted with one or more deuterium atoms, A 2 and R 2 Any one or both of these may be C1-C6 alkyl substituted with one or more deuteriums. For example, A 2 and R 2 Each may independently be a C1-C3 alkyl optionally substituted with deuterium. For example, Z 1 is N or CH, and A 2 and R 2 are each independently CH3 or CD3, A 2 and R 2 Either or both of these may be CD3.

[0057] In some embodiments, the compound of Formula I herein can be a compound represented by Formula IC:

[0058] [Chemical Formula IC]

[0059]

[0060] In the above chemical formula IC,

[0061] Z 1 is N or CH,

[0062] R 1is H; or C1-C3 alkyl optionally substituted with halogen,

[0063] R 2 is a straight or branched chain C1-C6 alkyl optionally substituted with one or more deuterium; or a C3-C7 cycloalkyl optionally substituted with one or more deuterium or halogen.

[0064] In one specific example of the above chemical formula IC, R 1 is C1-C3 alkyl substituted with H or halogen, and R 2 is a straight or branched C1-C3 alkyl optionally substituted with one or more deuteriums; or a C3-C5 cycloalkyl optionally substituted with one or more deuteriums or halogens. In this case, R 2 For example, R may be a straight or branched chain C1-C3 alkyl, a straight or branched chain C1-C3 alkyl substituted with one or more deuteriums, a C3-C5 cycloalkyl, a C3-C5 cycloalkyl substituted with one or more deuteriums, or a C3-C5 cycloalkyl substituted with one or more halogens. For example, R 1 is H or CF3, and R 2 can be CH3, CD3 or cyclopropyl. For example, R 1 is H, and R 2 may be CH3, CD3 or cyclopropyl. In one embodiment, the compound of formula I, formula IA, formula IB or formula IC of the present invention may be a compound selected from the group consisting of:

[0065]

[0066]

[0067]

[0068] In one specific embodiment, the compound of formula I, formula IA, formula IB or formula IC of the present invention may be a compound selected from the group consisting of:

[0069]

[0070] The present invention also includes a compound selected from the following, or a solvate, stereoisomer or pharmaceutically acceptable salt thereof.

[0071]

[0072] For example, the compound of the present invention may be the following compound:

[0073]

[0074] definition

[0075] All technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art, and unless otherwise stated, conventional measuring methods, manufacturing methods, conventional ingredients or materials based on conventional techniques of pharmacology, pharmaceutical manufacturing, mass spectrometry, NMR, HPLC, biochemistry, etc. are used.

[0076] The individual features and components of each embodiment described and illustrated in this specification may be combined with the features and components of any other embodiment without departing from the scope or spirit of the present disclosure.

[0077] Unless otherwise specified, in this specification and the appended claims, "or" and "and" mean "and / or." The terms "comprises" and "comprising" are open-ended, meaning that the compound, composition, or method may include additional features or components in addition to the specific features or components listed. Throughout the description and claims of this specification, the singular includes the plural unless the context requires otherwise. In particular, where the indefinite article "a" is used, it is to be understood that this specification contemplates the plural as well as the singular, unless the context requires otherwise.

[0078] In this specification, a numerical range indicated using the term "within" refers to a range that includes the numerical values ​​described before and after the term "within" as the lower and upper limits, respectively. In this specification, a numerical value indicated using the term "about" may refer to a range of ±20% of the corresponding numerical value, preferably ±10% of the corresponding numerical value.

[0079] As used herein, the terms "optional" or "optionally" mean that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, the term "optionally substituted" means that the occurrences include instances where the occurrences are either substituted or unsubstituted with the specified substituent.

[0080]

[0081] compound

[0082] The term "alkyl" as used herein refers to a fully saturated branched or unbranched (or straight-chain or linear) hydrocarbon. The alkyl may be a substituted or unsubstituted alkyl group. The term "alkyl" as used herein may refer to, for example, C1-C6 alkyl, C1-C3 alkyl, etc. The C1-C6 alkyl may be an alkyl group that is C1 to C6, C1 to C5, C1 to C4, C1 to C3, or C1 to C2. Non-limiting examples of the alkyl may be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, n-pentyl, isopentyl, neopentyl, iso-amyl, or n-hexyl.

[0083] As used herein, the term "cycloalkyl" refers to a saturated hydrocarbon ring having the specified number of carbon atoms as ring elements (i.e., C3-C6 cycloalkyl refers to a cycloalkyl group having 3, 4, 5, or 6 carbon atoms as ring elements). The term "cycloalkyl" as used herein may refer to, for example, C3-C6 cycloalkyl, C3-C5 cycloalkyl, or C3-C4 cycloalkyl. The cycloalkyl may be, for example, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. If the cycloalkyl contains one or more double bonds, it is referred to as a "partially unsaturated" cycloalkyl, but does not include an aryl ring.

[0084] The term "halogen" as used herein refers to an atom belonging to Group 17 of the periodic table. Halogen atoms include fluorine, chlorine, bromine, and iodine, and may be used interchangeably with the term "halo," which refers to a monovalent functional group composed of halogen.

[0085] The term "haloalkyl" as used herein includes branched and straight-chain saturated aliphatic hydrocarbon groups having a specific number of carbon atoms, substituted with one or more halogen atoms. Haloalkyl may include perhaloalkyl, which may refer to a functional group in which all hydrogens of the alkyl group are replaced by halogens (e.g., -CF3, -CF2CF3). The haloalkyl may include two or more halogens, and each halogen may be the same (e.g., CHF2, -CF3) or different (e.g., CF2Cl).

[0086] As used herein, the term "heterocyclyl" may be used interchangeably with "heterocyclic", "heterocycle" or "heterocycloalkyl" and, unless otherwise stated, refers to a monocyclic or polycyclic saturated or partially unsaturated ring system having the specified number of ring elements and containing one or more heteroatoms selected from B, N, O, S, Si and P. That is, a 3- to 7-membered heterocyclyl refers to a heterocyclyl group having 3, 4, 5, 6 or 7 ring elements, including the heteroatoms. One or more N or S atoms in a heterocyclyl group may be oxidized (e.g., N → O-, S(O), SO2), and ring systems containing oxidized N or S atoms may also be included in the heterocyclyl. The heterocyclyl may contain 1 to 4 heteroatoms, 1 to 3 heteroatoms, 1 or 2 heteroatoms, or 1 heteroatom selected from N, O, and S. For example, the heteroatom may be N, O, or S. The heterocyclyl herein may contain a heteroatom or heteroatom group selected from N, O, S, and SO2 within the ring. In addition, the heterocyclyl may contain 5 to 10, 4 to 7, 4, 5, or 6 ring atoms. For example, the heterocyclyl may include, but is not limited to, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, azetidinyl, pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, thietanyl, tetrahydrothiophenyl, 1,1-dioxidothietanyl, 1,1-eioxidotetrahydrothiophenyl, and the like.

[0087] The term "deuterium" as used herein refers to the isotope of hydrogen with a mass number of 2, referred to as heavy hydrogen or deuterium, or by its abbreviation "D". The terms "deuterated" and "substituted with deuterium" are used interchangeably to refer to a moiety or compound in which all or part of the hydrogens contained in the substituent are replaced with deuterium.

[0088] The term "substitution" in the term "substituted or unsubstituted" or "optionally substituted" refers to the introduction of a substituted hydrogen atom in place of another atomic group when forming a derivative by replacing one or more hydrogen atoms in an organic compound, and the term "substituent" refers to the introduced atomic group.

[0089] In this specification, when a combination of substituents is referred to as one group, for example, arylalkyl, cycloalkylalkyl, etc., it generally contains the atom attached to the end of the molecule as the last mentioned group.

[0090] In this specification, "-" is used to indicate the position at which a substituent is bonded to the remaining residue of the compound. For example, when - is displayed at the end of a substituent, it means that the end is bonded to the remaining residue of the compound. Furthermore, when two or more substituents are connected by "-", it means that the substituent immediately before "-" is bonded to the substitutable atom of the substituent immediately after "-".

[0091] The term "solvate" as used herein may refer to a compound of the present invention or a salt thereof that contains a stoichiometric or non-stoichiometric solvent bound by non-covalent intermolecular forces. Preferred solvents for this purpose may be volatile, non-toxic, and / or suitable for human administration. The solvent may be water, in which case the "solvate" is referred to as a "hydrate."

[0092] The term "stereoisomer" as used herein may mean a compound of the present invention or a salt thereof having the same chemical or molecular formula but different optically or sterically, and specifically may be a diastereomer, an enantiomer, or a geometric isomer.

[0093] In some embodiments, the compounds of the present invention may contain one or more asymmetric centers, and may be in the form of racemates, single enantiomers, mixtures of enantiomers, single diastereomers, mixtures of diastereomers, etc. In one embodiment, due to the nature or restricted rotation of the asymmetric center, the compounds of the present invention may exist in the form of enantiomers or diastereomers.

[0094] When two or more asymmetric centers are present in the compounds of the present invention, multiple diastereoisomers and enantiomers of the chemical structures disclosed herein may exist, and all such pure isomers, isolated isomers, partially pure isomers, or racemic mixtures are intended to fall within the scope of the present invention.

[0095] Purification of the above isomers and separation of the isomer mixture can be achieved by standard techniques known in the art. For example, a diastereomeric mixture can be separated into individual diastereoisomers by chromatographic processes or crystallization, and racemates can be separated into individual enantiomers by chiral phase chromatographic processes or resolution.

[0096] The compound of the present invention can be used in the form of a pharmaceutically acceptable salt derived from an inorganic acid or an organic acid, for example, the salt can be a salt derived from hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, acetic acid, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, mandelic acid, tartaric acid, citric acid, ascorbic acid, palmitic acid, maleic acid, hydroxymaleic acid, benzoic acid, hydroxybenzoic acid, phenylacetic acid, cinnamic acid, salicylic acid, methanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, or the like.

[0097] A pharmaceutically acceptable salt of the above compound can be prepared by dissolving the compound of the present invention in a water-miscible organic solvent, such as acetone, methanol, ethanol, or acetonitrile, adding an excess of an organic acid or an aqueous solution of an inorganic acid, and then precipitating or crystallizing the mixture. Subsequently, the solvent or the excess of acid is evaporated from the mixture, followed by drying to obtain an addition salt, or the precipitated salt can be prepared by suction filtration.

[0098] General method for preparing compounds

[0099] The compound according to the present invention can be prepared through chemical modifications well known to those skilled in the art of organic / medicinal chemistry, according to the representative methods illustrated below.

[0100] The following general reaction scheme is a general example of a representative method for preparing a compound of chemical formula I. A person skilled in the art will be able to easily prepare a compound of chemical formula I by appropriately selecting a starting material, reaction temperature, reaction conditions, catalyst, solvent, treatment method, etc. suitable for the desired compound based on the preparation method specifically disclosed in the examples of the present application.

[0101] In the following reaction formula, the designation of each substituent of the compound of formula I and intermediates is the same as the designation of the substituent at the corresponding position in formula I, unless otherwise specified. In addition, in the following reaction formula, R' represents an alkyl group, and Hal represents a halo group (preferably Br or Cl). For the convenience of explanation, each ring structure of the compound of formula I is referred to as the A-part, the B-part, the C-part, and the D-part as follows. The same variables are defined the same, and repeated definitions are omitted.

[0102]

[0103] In one aspect, the compound of formula I can be prepared according to the preparation method of the following reaction scheme I. The preparation method of reaction scheme I is a method of first combining part B and part C, and then sequentially combining part A and part D.

[0104] [Reaction Formula I]

[0105]

[0106] In step 1 of reaction scheme I, intermediate INT-ABC can be prepared by amide coupling reaction of intermediate INT-BC with an A-part intermediate in the presence of an appropriate base (e.g., DIEA). In addition to the coupling reagents (EDCI and HOBt) described in reaction scheme I, the amide coupling reagents can be appropriately changed as needed, and reaction conditions such as an appropriate reaction time and reaction temperature can be selected accordingly. The amide coupling reaction of step 1 can be performed at about 20°C to about 50°C for about 30 minutes to about 6 hours. For example, the amide coupling reaction of step 1 can be performed at about 25°C for about 2 hours to about 5 hours, or at about 40°C for about 30 hours to about 2 hours.

[0107] In step 2 of reaction scheme I, the intermediate INT-ABC and the organotin D-part intermediate can be subjected to Stille coupling in the presence of a suitable solvent (e.g., dioxane) and a Pd catalyst (e.g., Pd(PPh)3Cl2) to prepare the compound of formula I. The reaction of step 2 can be carried out at a temperature of about 80°C to about 120°C or about 90°C to about 100°C for about 5 hours to about 30 hours or about 5 hours to about 20 hours. For example, it can be carried out at about 100°C for about 12 hours.

[0108] The intermediate INT-BC used in reaction scheme I is Z 1 In this case, N can be manufactured, for example, according to the following reaction scheme IA.

[0109] [Reaction Scheme IA]

[0110]

[0111] In step 1 of Scheme IA, the B-part intermediate and the C-part boronic acid ester intermediate can be coupled in the presence of a Cu(II) catalyst (e.g., Cu(OAc)2) using suitable reagents and solvents (e.g., boronic acid, molecular sieves, MeCN). The reaction of step 1 can be carried out at a temperature of about 80°C to about 120°C or about 90°C to about 100°C for about 5 hours to about 30 hours or about 5 hours to about 20 hours. For example, it can be carried out at about 90°C for about 16 hours.

[0112] In step 2 of reaction scheme IA, the ester group of the intermediate obtained in step 1 can be hydrolyzed under appropriate reaction conditions to produce intermediate INT-BC. For example, the hydrolysis reaction can be performed using NaOH in THF and H2O.

[0113] Alternatively, the intermediate INT-BC is Z 1In this case, CH can be manufactured, for example, according to the following reaction scheme IB.

[0114] [Reaction Formula IB]

[0115]

[0116] In step 1 of Scheme IB, a pyranone ester compound can be condensed with a C-part aminopyridine intermediate in the presence of a suitable base and solvent (e.g., pyridine). The reaction in step 1 can be carried out at a temperature of about 50°C to about 100°C or about 60°C to about 90°C for about 5 hours to about 30 hours or about 5 hours to about 20 hours. For example, it can be carried out at about 80°C for about 16 hours.

[0117] In step 2 of reaction scheme IB, intermediate INT-BC can be prepared through a hydrolysis reaction in the same manner as step 2 of reaction scheme IA.

[0118] Alternatively, a compound of formula I, such as R 2 The compound of formula I, which is a C3-C7 cycloalkyl substituted with one or more halogens, can be prepared by the method of the following reaction scheme I-1 using the intermediate INT-ABC.

[0119] [Reaction Scheme I-1]

[0120]

[0121] In Scheme I-1, the compound of formula I can be prepared by coupling the intermediate INT-ABC with the D-part intermediate in the presence of a suitable catalyst (e.g., a Pd catalyst such as Pd(OAc)2 and Xphos), a base (e.g., K2CO3) and a solvent (e.g., DMF). The reaction of Scheme I-1 can be carried out at a temperature of about 90°C to about 140°C or about 100°C to about 120°C for about 10 hours to about 30 hours or about 15 hours to about 20 hours. For example, it can be carried out at about 110°C for about 16 hours.

[0122] Alternatively, a compound of formula I, such as R 2 The compound of formula I, which is a 3- to 7-membered heterocyclyl containing S or SO2, can be prepared using the intermediate INT-ABC according to the method of the following reaction scheme I-2.

[0123] [Reaction Scheme I-2]

[0124]

[0125] Step 1 of Scheme I-2 can be carried out in a similar manner to Step 2 of Scheme I using a triazole compound substituted with S-containing heterocyclyl (ring BB) as a D-part intermediate. According to Step 1 of Scheme I-2, R 2 A compound of formula I, which is a 3- to 7-membered heterocyclyl containing S, can be prepared.

[0126] R of the compound of formula I 2When the compound is a 3- to 7-membered heterocyclyl group containing a sulfone (SO2), the intermediate obtained in step 1 of the above reaction scheme I-2 can be further oxidized to prepare a compound of formula I. Step 2 of the reaction scheme I-2 can be performed, for example, using oxone in an appropriate solvent (e.g., THF and H2O). The reaction of step 2 can be performed at a temperature of about 20°C to about 40°C or about 20°C to about 30°C, for about 5 hours to about 30 hours or about 5 hours to about 20 hours. For example, it can be performed at about 25°C for about 12 hours.

[0127] In another aspect, the compound of formula I can be prepared according to the preparation method of the following reaction scheme II.

[0128] [Reaction Formula II]

[0129]

[0130] The above reaction scheme II is a manufacturing method in which the reaction order is changed in steps 1 and 2 of reaction scheme I, and step 2 of reaction scheme IA or reaction scheme IB. That is, the manufacturing method of reaction scheme II is a method in which B-part and C-part are combined first, and D-part and A-part are combined sequentially.

[0131] Specifically, steps 1 and 3 of Scheme II can be performed in the same or similar manner as steps 2 and 1 of Scheme 1, respectively. Step 2 of Scheme II can be performed in the same or similar manner as step 2 of Scheme IA or IB.

[0132] In another aspect, the compound of formula I can be prepared according to the following reaction scheme III. The preparation method of reaction scheme III is a method of first combining the C-part and the D-part, and then sequentially combining the B-part and the A-part.

[0133] [Reaction Formula III]

[0134]

[0135] In step 1 of the above reaction scheme III, the boronic acid intermediate INT-CD can be coupled with the B-part intermediate using a suitable base (e.g., TEA and pyridine) and a solvent (e.g., MeCN) in the presence of a Cu(II) catalyst (e.g., Cu(OAc)2). The reaction of step 1 can be carried out at a temperature of about 80°C to about 120°C or about 90°C to about 100°C for about 5 hours to about 30 hours or about 5 hours to about 20 hours. For example, it can be carried out at about 90°C for about 16 hours.

[0136] Steps 2 and 3 of Scheme III can be performed in the same manner as steps 2 and 3 of Scheme II.

[0137] The boronic acid intermediate INT-CD used in the above reaction scheme III can be prepared according to the following reaction scheme IIIA.

[0138] [Reaction Scheme IIIA]

[0139]

[0140] In step 1 of the above reaction scheme IIIA, the halogenated pyridine compound and the organotin D-part intermediate can be Stille coupled in a manner similar to step 2 of the reaction scheme I. The reaction of step 1 can be carried out at a temperature of about 80°C to about 120°C or about 90°C to about 100°C for about 5 hours to about 30 hours or about 5 hours to about 20 hours. For example, it can be carried out at about 90°C for about 16 hours.

[0141] In step 2 of the above reaction scheme IIIA, the halogenated pyridine compound obtained in step 1 can be borated using a suitable boronating agent (e.g., bis(pinacolato)diboron) and a Pd(II) catalyst (e.g., Pd(dppf)Cl2) in the presence of a suitable base (e.g., KOAc) and a solvent (e.g., dioxane) to produce a boronic acid intermediate INT-CD. The reaction in step 2 can be carried out at a temperature of about 80°C to about 120°C or about 90°C to about 100°C for about 5 hours to about 30 hours or about 5 hours to about 20 hours. For example, it can be carried out at about 100°C for about 16 hours.

[0142]

[0143] Medicinal uses, pharmaceutical compositions and methods of administration

[0144] The compounds of the present invention, including the compound of the above formula I, stereoisomers, solvates, or pharmaceutically acceptable salts thereof, can be used to prevent or treat SOS1-mediated diseases. The compounds of the present invention, including the compound of the above formula I, stereoisomers, solvates, and pharmaceutically acceptable salts are as described above.

[0145] As used herein, the term "preventing" or "prevention" refers to preventing a disease, condition or disorder, for example, in an individual who may be predisposed to the disease, condition or disorder but who has not yet experienced or exhibited the pathology or signs of the disease.

[0146] As used herein, the term "treating" or "treatment" includes inhibiting a disease, condition or disorder, e.g., inhibiting the disease, condition or disorder in a subject experiencing or exhibiting the pathology or signs of the disease, condition or disorder, i.e., preventing recurrence or further development of the pathology and / or signs after treatment of the pathology and / or signs, or ameliorating a disease, condition or disorder, e.g., ameliorating the disease, condition or disorder in a subject experiencing or exhibiting the pathology or signs of the disease, condition or disorder, i.e., reversing the pathology and / or signs, e.g., reducing disease severity.

[0147] The above SOS1-mediated disease may include a disease that can be prevented or treated by inhibiting the interaction between SOS1 and a RAS family protein, and / or SOS1 and RAC1. The SOS1-mediated disease may include a disease associated with abnormal activity of SOS1 and / or a RAS family protein. The SOS1-mediated disease may be, for example, cancer. The cancer may be, for example, pancreatic cancer, lung cancer, colorectal cancer, cholangiocarcinoma, multiple myeloma, melanoma, uterine cancer, cervical cancer, endometrial cancer, thyroid cancer, chronic lymphocytic leukemia, acute myeloid leukemia, bladder cancer, urothelial cancer, gastric cancer, head and neck squamous cell carcinoma, diffuse large B-cell lymphoma, esophageal cancer, hepatocellular carcinoma, breast cancer, ovarian cancer, prostate cancer, glioblastoma, renal cancer, or sarcoma. In one specific example, the cancer may be pancreatic cancer, lung cancer (e.g., non-small cell lung cancer), biliary tract cancer, or colorectal cancer.

[0148] The cancer may be, for example, a cancer dependent on the RAS family and MAPK signaling pathway. Such cancers may include, for example, cancers exhibiting mutations, gene amplification and / or overexpression of proteins or genes in the RAS family and MAPK signaling pathway (e.g., mutations, amplification or overexpression of RAF, MEK), such as KRAS, NRAS, HRAS, receptor tyrosine kinases (e.g., EGFR, ErbB2, ErbB3, ErbB4, PDGFR-A / B, FGFR1 / 2 / 3, IGF1R, INSR, ALK, ROS, TrkA, TrkB, TrkC, RET, c-MET, VEGFR1 / 2 / 3, AXL), GAP (e.g., NF1) and SOS1. In addition, the cancer may be a RAC1-dependent cancer.

[0149] SOS1-mediated diseases may be, for example, diseases associated with dysregulation of the RAS family protein pathway, i.e., RASopathies. Such RASopathies may include Neurofibromatosis type 1 (NF1), Noonan Syndrome, Noonan Syndrome with Multiple Lentigines (NSML), also called Leopard Syndrome, Capillary Malformation-Arteriovenous Malformation Syndrome (CM-AVM), Costello Syndrome, Cardio-Facio-Cutaneous Syndrome (CFC Syndrome), Legius Syndrome (also called NF1-like syndrome), or Hereditary gingival fibromatosis.

[0150] In one embodiment, the compound of the present invention, including the compound of formula I, can be used to treat diseases associated with abnormal activity of SOS1 or RAS family proteins, or abnormal regulation of pathways of RAS family proteins, by inhibiting the interaction between SOS1 and RAS family proteins, or SOS1 and RAC1.

[0151] The compounds of the present invention, including the compound of formula I, significantly inhibit the interaction between SOS1 and RAS family proteins, or SOS1 and RAC1, and exhibit excellent anticancer efficacy and RAS disease treatment efficacy. In addition, the compounds of the present invention, including the compound of formula I, exhibit optimal drug metabolism and pharmacokinetics (DMPK) characteristics and good PK exposure. Furthermore, they exhibit time-dependent inhibition of CYP3A4 as well as a reduced risk of induction, and thus can be used clinically without concerns about drug interactions.

[0152] In one embodiment, the pharmaceutical composition may include a conventional pharmaceutically acceptable carrier, excipient, or additive. The pharmaceutical composition may be formulated according to a conventional method and may be prepared in various oral dosage forms such as tablets, pills, powders, capsules, syrups, emulsions, microemulsions, etc., or in parenteral dosage forms such as intramuscular, intravenous, or subcutaneous administration. The pharmaceutical composition may be a single composition or separate compositions. The pharmaceutical composition includes a compound, stereoisomer, solvate, or pharmaceutically acceptable salt according to one aspect as an active ingredient of the pharmaceutical composition.

[0153] When the pharmaceutical composition of the present invention is manufactured in the form of an oral dosage form, examples of additives or carriers used include cellulose, calcium silicate, corn starch, lactose, sucrose, dextrose, calcium phosphate, stearic acid, magnesium stearate, calcium stearate, gelatin, talc, surfactants, suspending agents, emulsifiers, diluents, etc. When the pharmaceutical composition of the present invention is manufactured in the form of an injection, examples of additives or carriers include water, saline solution, glucose aqueous solution, pseudo-sugar aqueous solution, alcohol, glycol, ether (e.g., polyethylene glycol 400), oil, fatty acid, fatty acid ester, glyceride, surfactant, suspending agent, emulsifier, etc.

[0154] The dosage of the pharmaceutical composition is an amount effective for the treatment or prevention of an individual or patient, and may be administered orally or parenterally, depending on the purpose. For oral administration, the dosage is 0.01 to 1000 mg, more specifically 0.1 to 300 mg per kg of body weight per day, based on the active ingredient, and for parenteral administration, the dosage is 0.01 to 100 mg, more specifically 0.1 to 50 mg per kg of body weight per day, based on the active ingredient, and may be administered once or in several divided doses. The dosage for a specific individual or patient should be determined in light of various related factors such as the patient's weight, age, sex, health condition, diet, administration time, administration method, and severity of the disease, and it should be understood that it can be appropriately increased or decreased by a specialist, and the above dosage is not intended to limit the scope of the present invention in any way. A physician or veterinarian having ordinary skill in the relevant art can easily determine and prescribe an effective amount of the required pharmaceutical composition. For example, a physician or veterinarian may start the dosage of a compound of the present invention used in a pharmaceutical composition at a level lower than that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.

[0155] In one embodiment, the pharmaceutical composition includes within its scope a pharmaceutical composition comprising, as an active ingredient, a therapeutically effective amount of at least one compound according to one embodiment, alone or in combination with a pharmaceutical carrier. The term "therapeutically effective amount" or "effective amount" means an amount sufficient to produce a beneficial or desired clinical result, e.g., an amount sufficient to alleviate, improve, stabilize, reverse, slow, or delay the progression of a disease.

[0156] Meanwhile, as described below, when the compound disclosed herein is administered in combination with one or more other therapeutic agents, a pharmaceutical composition comprising the compound according to the present invention and the other therapeutic agents is provided herein. Accordingly, in one specific embodiment, a pharmaceutical composition for preventing or treating cancer is provided, comprising a compound of the present invention, including a compound of formula I, or a solvate, stereoisomer, or pharmaceutically acceptable salt thereof; and an anticancer agent as active ingredients. The compound of the present invention, including the compound of formula I, and the anticancer agent may be included in one pharmaceutical composition, or each may be formulated as separate pharmaceutical compositions. The anticancer agent to be used in combination is as described below.

[0157] Combination administration

[0158] Optionally, the compound according to one embodiment may be administered alone, in combination with a compound according to another embodiment, or concurrently, separately, or sequentially with one or more other therapeutic agents, e.g., anticancer agents or other pharmaceutically active substances.

[0159] When used for cancer treatment, the compound of the present invention may be administered in combination with other anticancer agents. Accordingly, one aspect of the present invention provides a pharmaceutical composition for preventing or treating cancer, comprising a compound according to the present invention, a solvate, stereoisomer, or pharmaceutically acceptable salt thereof; and an anticancer agent as an active ingredient. In addition, one aspect of the present invention provides a method for preventing or treating cancer, comprising the steps of administering a compound according to the present invention, a solvate, stereoisomer, or pharmaceutically acceptable salt thereof, to a subject in need thereof; and simultaneously, separately, or sequentially administering an anticancer agent to the subject.

[0160] The anticancer agent that can be administered in combination with the compound of the present invention may be selected from the group consisting of chemotherapeutic agents, targeted anticancer agents, anticancer viruses, antibody therapeutic agents, immunocytotherapy agents, immune checkpoint inhibitors, and combinations thereof.

[0161] In one embodiment, the compound of the present invention is used alone or in combination with other anticancer therapies, such as radiotherapy, taxane derivatives (e.g., paclitaxel, docetaxel), platinum compounds (e.g., cisplatin, carboplatin), antimetabolites (e.g., 5-FU, gemcitabine, cytarabine, 6-thioguanine), CDK4 / 6 inhibitors (e.g., abemaciclib, palbociclib), immunotherapeutic agents (e.g., anti-CTLA4 antibodies, anti-PD1 antibodies), angiogenesis inhibitors (e.g., bevacizumab, nintedanib, regorafenib), topoisomerase inhibitors (e.g., irinotecan, SN-38, doxorubicin), ERK inhibitors (e.g., ulixertinib, lineterkip), MDM2 inhibitors (e.g., alizomadlin), PARP inhibitors (niraparib), MCL-1 inhibitors, mTOR inhibitors (e.g., rapamycin, temsirolimus, INK-128 (sapanisertib), everolimus), BET inhibitors (JQ1), CDK9 inhibitors (e.g., AT-7519), IGF1 / 2 or IGF1-R inhibitors (NVP-ADW742), PIK inhibitors (duvelisib), EGFR inhibitors (e.g., afatinib, osimertinib, cetuximab, lazertinib, zongetinib, gefitinib, neratinib), ErbB2 (HER2) inhibitors (e.g., trastuzumab, irbinitinib, neratinib), ALK inhibitors (e.g., crizotinib, alectinib), MEK inhibitors (e.g., trametinib, cobimetinib), BCR-ABL inhibitors (e.g., imatinib, nilotinib, dasatinib), FGFR1, FGFR2 or FGFR3 inhibitors (e.g., nintedanib), ROS1 inhibitors (e.g., crizotinib, entrectinib, repotrectinib), c-MET inhibitors (e.g., tepotinib), AXL inhibitors (e.g., bemcentinib), NTRK1 inhibitors (e.g., repotrectinib), RET inhibitors (e.g., pralsetinib), KRAS G12C inhibitors (e.g., sotorasib, adagrasib, dibarasib, trametinib, JDQ443), KRAS G12D inhibitors (e.g., MRTX1133), SHP2 inhibitors (e.g., TNO155), mutBRAF inhibitors (e.g., dabrafenib),PI3K inhibitors (e.g., alpelisib, apitolisib), Aurora A inhibitors (e.g., MK-5108), pan Aurora inhibitors (e.g., danucertib, AMG-900, reversine), BTK inhibitors (e.g., ibrutinib), Wee1 inhibitors (e.g., MK-1775), DHFR inhibitors (e.g., methotrexate), HSP90 inhibitors (e.g., BIIB021), A3AR antagonists (e.g., reversine), ubiquitin E1 enzyme inhibitors (e.g., MLN-7243), Bcl-2 inhibitors (e.g., venetoclax, ABT-737), EZH2 inhibitors (e.g., GSK-343), ARID1A inhibitors (e.g., GSK-343), SUMOylation inhibitors (e.g., 2-D08), Chk1 inhibitors (e.g., SCH-900776), HDAC inhibitors (e.g., entinostat), JAK1 / 2 inhibitors (e.g., ruxolitinib), proteasome inhibitors (e.g., carfilzomib), Akt inhibitors (e.g., ipatasertib), GR inhibitors (e.g., prednisolone), PLK1 inhibitors (e.g., volasertib), pan-RAF inhibitors (e.g., sorafenib), RAS-RAF-MEK-ERK signaling inhibitors (e.g., abutometinib), etc.

[0162] The compound of the present invention can be used in combination with other anticancer therapies described above to enhance the effects of the anticancer therapies, thereby significantly inhibiting the expression and activity of target proteins or genes. Specifically, the compound of the present invention not only possesses its own anticancer efficacy, but can also enhance the anticancer efficacy of the target inhibitor. Therefore, when the compound of the present invention is used in combination with the anticancer therapies, excellent anticancer efficacy can be achieved.

[0163] As used herein, the term "chemotherapeutic agent" is also referred to as an antitumor drug or cytotoxic agent. It is a general term for drugs that exhibit anticancer activity primarily by directly acting on DNA to block DNA replication, transcription, and translation processes, interfering with the synthesis of nucleic acid precursors in metabolic pathways, and inhibiting cell division. These antitumor drugs exhibit cytotoxicity not only on tumor cells but also on normal cells. Chemotherapeutic agents can be used for maintenance therapy. Furthermore, the term "maintenance therapy" as used herein refers to a treatment method implemented to prevent or delay cancer recurrence by treating cancer with drugs after initial anticancer treatment.

[0164] Specifically, the chemotherapeutic agent may be any one selected from the group consisting of an alkylating agent, a microtubule inhibitor, an antimetabolite, and a topoisomerase inhibitor. The alkylating agent may be any one selected from the group consisting of mechlorethamine, cyclophosphamide, ifosfamide, melphalan, chlorambucil, thiotepa, altretamine, procarbazine, busulfan, streptozotocin, carmustine, lomustine, dacarbazine, cisplatin, carboplatin, and oxaliplatin. The microtubule inhibitor may be any one selected from the group consisting of docetaxel, paclitaxel, velban, oncovin, and navelbine. The anti-metabolite may be any one selected from the group consisting of Fluorouracil, Capecitabine, Cytarabine, Gemcitabine, Fludarabine, Methotrexate, Pemetrexed, 6-thioguanine, and Mercaptopurine. The topoisomerase inhibitor may be any one selected from the group consisting of Hycamtin, Camptosar, Vepesid, Blenoxane, Adriamycin, SN-38, Doxorubicin, and Cerubidine.

[0165] As used herein, the term "targeted anticancer agent" refers to a therapeutic agent that specifically kills cancer cells by targeting specific proteins or genetic changes that are abundant only in cancer cells and blocking signals involved in cancer growth and development. These agents are categorized as monoclonal antibodies that react outside the cell and small molecules that act inside the cell. Monoclonal antibodies are anticancer agents that block cancer cell-inducing signals transmitted outside the cell, acting on initiating signals related to proliferation and apoptosis, while small molecules act on complex signaling that occurs inside the cell.

[0166] Specifically, the targeted proteins are mTOR, PI3K, EGFR, VEGFR, CD20, CD38, RNAK-L, BTK, Bcr-abl, PDGFR / FGFR family, MEK, KRAS, ERK1 / 2, HER2 / Neu, Ubiquitin, JAK, ALK, PARP, TGFβR, Proteasome, Bcl-2, C-Met, VR1, VR2, VR3, c-kit, AXL, RET, BRAF, pan-RAF, SHP2, SRC, LCK, DNMT, CDK4 / 6, CDK9, BET, MDM2, IGF1 / 2 or IGF1-R, ROS1, NTRK1, PIK, DHFR, pan Aurora, Aurora A, WEE1, HSP90, A3AR, EZH2, ARID1A, Chk1, ATR, HDAC1 / 3, These could be Akt, PLK1, SUMOylation-related proteins, STING, etc.

[0167] The above targeted anticancer drugs are Rapamycin, Sirolimus, Temsilorimus, Everolimus, Ridaforolimus, INK-128, Alpelisib, Cetuximab, Trastuzumab, Pertuzumab, Gefitinib, Erlotinib, Osimertinib, Lazertinib, Zongertinib, Panitumumab, Axitinib, Lenvatinib, Bevacizumab, Ramucirumab, Aflibercept, Rituximab, Obinutuzumab, Daratumumab, Denosumab, Ibrutinib, Dasatinib, Nilotinib, Imatinib, Bosutinib, Galunisertib, Vactosertib, Futibatinib, Nintedanib, Sunitinib, Sorafenib, Avutometinib, Cabozantinib, Regorafenib, Masitinib, Semaxanib, Tivozanib, Vandetanib, Pazopanib, Dabrafenib, Sotorasib, Adagrasib, Divarasib, JDQ443, MRTX1133, Ulixertinib, Afatinib, Lapatinib, Neratinib, Lenalidomide, Ixazomib, Ruxolitinib, Lestaurtinib, Pacritinib, Trametinib, Cobimetinib, Selumetinib, Binimetinib, Alectinib, Lorlatinib, Crizotinib, Venetoclax, Bemcentinib, Gilteritinib, Selpercatinib, Pralcetinib, Encorafenib, Vemurafenib, Belvarafenib, RMC-4630, Batoprotafib, WH-4-023, Olaparib, Talazoparib, Niraparib, Rucaparib, Azacitidine,Decitabine, Guadecitabine, Abemaciclib, Ribociclib, Palbociclib, CDNs, SB11285, Rineterkib, Repotrectinib, Tepotinib, Alrizomadlin, JQ1, NVP-ADW742, Duvelisib, Irbinitinib, Danusertib, MK-1775, AMG-900, BIIB021, Reversine, MLN-7243, ABT-737, MK-5108, GSK-343, 2-D08, SCH-900776, Entinostat, Carfilzomib, Apitolisib, Ipatasertib, Volasertib, AT-7519, Methotrexate, Wortmannin, ERAS-007, PYR-41, MLN4924, RO-5503781, MK-8242, SAR-405838, CGM097, DS3032b, Lactacystin, Disulfiram, Epigallocatechin-3-gallate, Marizomib, Oprozomib, Delanzomib, Epoxomicin, MG132, Beta-hydroxy beta-methylbutyrate, Bortezomib, Navitoclax, Naporafenib, PF-07284892, TNO155, Hesperadin, LY3295668 , Tozasertib, Azenosertib, ZNL-02-096, RP-6306, GSK-1520489A, BIIB028, MPC-3100, PU-H71, Debio093, SNX-5422, AUY922, KF-26777, MRS-545, CAY10498, DZNep, EPZ005687, EI1, GSK126, UNC1999, Tazemetostat, Sinefungin, GSK-343, Davidiin, CID9549553, SRA737, V158411, PF-477736, AZD7762, Prexasertib, Berzosertib, Gartisertib,Ceralasertib, Panobinostat, Mocetinostat, Trichostatin A, CBUD-1001, Abexinostat, VQD-002, Perifosine, Miltefosine, MK-2206, AZD5363, Rigosertib, I-BET 151, I-BET 762, OTX-015, TEN-010, CPI-203, CPI-0610, Olinone, RVX-208, ABBV-744, LY294002, AZD5153, MT-1, MS645, Figitumumab, Mecasermin, rhIGF-1, BI 885578, Buparlisib, Copanlisib, Dactolisib, Idelalisib, Parsaclisib, Paxalisib, Taselisib, Zandelisib, It may be any one selected from the group consisting of Inavolisib, AZD4573, Atuveciclib, VIP152, A-1592668, JSH-150, SLS009, Roscovitine, and DMXAA.

[0168] In one embodiment, the compound of the present disclosure can be administered in combination with a targeted anticancer agent that acts on one or more targets selected from Bcl-2, EGFR, KRAS, MEK, and RAF. For example, the compound of the present disclosure can be administered in combination with a targeted anticancer agent selected from venetoclax, zongetinib, osimertinib, lazertinib, cetuximab, sotorasib, adagrasib, dibarasib, MRTX1133, trametinib, and abutometinib. As used herein, the term "mTOR (mammalian target of rapamycin)" is also called mechanistic target of rapamycin or FRAP1 (FK506 binding protein 12-rapamycin associated protein 1), which is a protein belonging to the PIKK (phosphatidylinositol 3-kinase-related kinase) family. mTOR, encoded by the FRAP1 gene in humans, is a serine / threonine protein kinase that regulates cell growth, proliferation, motility, survival, protein synthesis, and transcription. mTOR inhibitors can suppress tumor survival by inhibiting autophagy, adipogenesis, proliferation, and protein synthesis. Examples of mTOR inhibitors include rapamycin, sirolimus, temsilorimus, everolimus, ridaforolimus, and INK-128 (Sapanisertib, MLN0128, TAK-228).

[0169] In this specification, "PI3K (Phosphoinositide 3-kinase)" is also called phosphatidylinositol 3-kinase, and is an enzyme involved in cellular functions such as cell growth, proliferation, differentiation, motility, survival, and intracellular signal regulation, and is related to cancer. PI3K contains subunits such as p110-α / β / γ / δ (PI3Kα / β / γ / δ). PI3K targeting anticancer drugs may include Alpelisib, Wortmannin, LY294002, Idelalisib, Copanlisib, Duvelisib, Apitolisib (GDC-0980, RG7422, GNE 390), etc.

[0170] As used herein, the term "epidermal growth factor receptor (EGFR)" refers to a cell membrane receptor that regulates cell growth, division, survival, and apoptosis. EGFR expression is increased in tumor tissues of various cancers. Tumor tissues with increased EGFR are known to have high invasion, metastasis, and anticancer drug resistance. As an EGFR inhibitor, a substance that inhibits EGFR may be, for example, Cetuximab, Trastuzumab, Pertuzumab, Gefitinib, Erlotinib, Osimertinib, Lazertinib, Zongertinib, or Panitumumab.

[0171] As used herein, the term "Vascular Endothelial Growth Factor Receptor (VEGFR)" refers to a cell membrane receptor of an angiogenic factor that induces angiogenesis, and a VEGFR inhibitor inhibits angiogenesis, thereby suppressing tumor growth and metastasis. A specific example of a VEGF inhibitor or VEGFR inhibitor may be Axitinib, Lenvatinib, Bevacizumab, Ramucirumab, or Aflibercept.

[0172] In this specification, the term "CD20 (B lymphocyte antigen CD20)" refers to a protein expressed on the surface of B cells and is used as a target protein for treating B cell lymphoma. The CD20 target inhibitor may be Rituximab or Obinutuzumab.

[0173] In this specification, the term "CD38 (Cluster of differentiation 38)" refers to a protein that acts as a signal transduction receptor in immune cells and regulates cell proliferation and death, and an inhibitor targeting this may be Daratumumab.

[0174] As used herein, the term "RNAK-L (Receptor activator of nuclear factor kappa-B ligand)" refers to the RANK receptor expressed on the surface of osteoclasts, which, when activated by binding to a ligand, causes bone destruction. RANK-L inhibitors are primarily used for cancer patients suffering from bone metastasis or osteoporosis, and may be specifically Denosumab.

[0175] In this specification, the term "BTK (Bruton's tyrosine kinase)" refers to an enzyme involved in the proliferation of B cells, which, when overexpressed, can develop into blood cancer. A specific example of a BTK target inhibitor may be Ibrutinib.

[0176] As used herein, the term "Bcr-abl" refers to a fusion protein highly expressed in patients with chronic myeloid leukemia, known to induce abnormal proliferation of blood cells. Specifically, the inhibitor of the protein may be Dasatinib, Nilotinib, Imatinib, or Bosutinib.

[0177] As used herein, the term "tumor growth factor β receptor (TGFβR)" refers to a cell membrane receptor of a tumor growth factor, which regulates the growth, migration, differentiation, and apoptosis of epithelial cells and hematopoietic cells. Examples of the TGFβR target inhibitor include, but are not limited to, Galunisertib or Vactosertib.

[0178] In this specification, the term "PDGFR (Platelet derived growth factor)" refers to a cell membrane receptor of PDGF, which is frequently expressed in cancer cells, and is known to regulate cancer growth, metastasis, and drug resistance by participating in angiogenesis. Fibroblast growth factor receptor (FGFR) is a receptor for fibroblast growth factor (FGF) and regulates various biological processes including cell growth, differentiation, and migration. The FGFR gene is prone to mutation, and such variants are commonly observed in breast cancer, uterine cancer, ovarian cancer, and cervical cancer. Inhibitors targeting PDGFR or FGFR may be Futibatinib, Nintedanib, Sunitinib, Imatinib, Sorafenib, Cabozantinib, Lenvatinib, Regorafenib, Masitinib, Semaxanib, Tivozanib, Vandetanib, Axitinib, or Pazopanib.

[0179] As used herein, the term "MEK (Mitogen-activated protein kinase kinase)" refers to a dual-specificity kinase enzyme that phosphorylates MAPK (mitogen-activated protein kinase), also called MAP2K, MEK, or MAPKK, and inhibition of MEK blocks cell proliferation and induces cell death. The MEK-targeting anticancer agent may be Cobimetinib, Selumetinib, Trametinib, or Binimetinib.

[0180] As used herein, the term "KRAS (Kirsten rat sarcoma virus)" refers to a gene that encodes a protein called K-Ras, which is part of the RAS / MAPK pathway and is an oncogene that directs cell growth, division, proliferation, and differentiation signals. KRAS inhibitors can target mutations such as KRAS G12C and KRAS G12D. KRAS-targeted anticancer agents may be Sotorasib, Adagrasib, Divarasib, JDQ443, or MRTX1133.

[0181] In this specification, the term "ERK1 / 2 (extracellular signal-regulated kinases 1 / 2)" refers to a widely expressed protein kinase intracellular signaling molecule involved in functions including regulation of meiosis, mitosis, and post-mitotic functions in cells, and disruption of the ERK pathway is commonly observed in cancer. The ERK1 / 2 targeted anticancer agent may be Rineterkib, Ulixertinib (BVD-523), or ERAS-007.

[0182] In this specification, the term "HER-2 / neu (Human epidermal growth factor receptor 2) regulates cell proliferation by activating PI3K / AkT. It is overexpressed in metastatic breast cancer and ovarian cancer, etc., and is known to induce anticancer drug resistance. The Her2 / neu targeted anticancer agent may be Trastuzumab, Afatinib, Lapatinib, Irbinitinib (Tucatinib), or Neratinib.

[0183] As used herein, the term "ubiquitin" refers to a protein that binds to other proteins and induces protein degradation by the proteasome, a protein-degrading enzyme (ubiquitin-proteasome system, UPS), thereby maintaining cellular homeostasis. Abnormal expression or activity of the UPS is observed in various tumors, and inhibitors thereof exhibit anticancer activity. For example, ubiquitin E1 enzyme target inhibitors may include MLN-7243 (TAK-243), PYR-41, MLN4924, etc., and MDM2 E3 ubiquitin ligase inhibitors may include RO-5503781 (Idasanutlin), MK-8242, SAR-405838, CGM097, DS3032b, etc.

[0184] As used herein, a "proteasome inhibitor" can treat cancer by blocking the action of the proteasome, a cellular complex that degrades proteins. Proteasome inhibition prevents the degradation of pro-apoptotic factors such as the p53 protein, thereby activating programmed cell death in tumor cells that depends on inhibition of the pro-apoptotic pathway. Proteasome inhibitors may include Lactacystin, Disulfiram, Epigallocatechin-3-gallate, Marizomib (salinosporamide A), Oprozomib (ONX-0912), Delanzomib (CEP-18770), Epoxomicin, MG132, Beta-hydroxy beta-methylbutyrate, Bortezomib, Carfilzomib, Ixazomib, and the like.

[0185] In this specification, the term "JAK (Janus kinase)" refers to a protein above STAT, a transcription factor that regulates cell proliferation, cell survival, cell migration, and immune response, and JAK inhibitors are known to reduce cell proliferation and induce apoptosis by inhibiting the activity of STAT. JAK includes JAK1, JAK2, JAK3, and TYK2 (tyrosine kinase 2). The JAK target inhibitor may be Ruxolitinib, Lestaurtinib, or Pacritinib.

[0186] As used herein, the term "ALK (Anaplastic lymphoma kinase)" refers to a signal transduction mediator that promotes cell proliferation, cell migration, angiogenesis, and inhibits apoptosis, and is overactivated in various cancer tissues. The ALK target inhibitor may be Alectinib, Lorlatinib, or Crizotinib.

[0187] As used herein, the term "Bcl-2" refers to a protein that inhibits cell death and is overexpressed or overactivated in various cancer tissues. Inhibitors targeting Bcl-2 may include Venetoclax, ABT-737, Navitoclax (ABT-263), and the like.

[0188] As used herein, the term "C-Met" refers to a receptor for hepatocyte growth factor (HGF), which activates signaling involved in cell growth, formation, motility, survival, and angiogenesis. The C-Met targeted anticancer agent may be Crizotinib, Tepotinib, or Cabozantinib.

[0189] In this specification, the term "VR (Vanilloid receptor)" is also known as TRPV (Transient receptor potential vanilloid) and exists in the form of VR1, VR2, VR3, VR4, VR5 and VR6. VR is known to regulate the proliferation, death, migration, invasion and angiogenesis of cancer cells at each stage in the cancer progression process.

[0190] As used herein, the term "c-kit," also known as CD117, induces signaling that activates cell survival, proliferation, and differentiation. c-kit is a proto-oncogene, and its overexpression or mutation is associated with cancer development. Specific examples of c-kit-targeting anticancer agents include Imatinib, Dasatinib, and Regorafenib.

[0191] As used herein, the term "AXL (Yyrosine-protein kinase receptor UFO)" refers to a cell surface tyrosine kinase receptor that mediates signal transduction involved in cell proliferation and survival. It is known to be involved in anticancer drug resistance in cancer treatment. A specific example of an AXL-targeting anticancer agent may be Bemcentinib or Gilteritinib.

[0192] In this specification, the term "RET (Rearragned during transfection)" refers to a receptor that mediates signals involved in cell proliferation, apoptosis, and survival, and mutations in RET are known to be involved in the development of cancer. The RET target inhibitor may be, but is not limited to, Selpercatinib or Pralsetinib.

[0193] As used herein, the term "BRAF" refers to a MAPK signaling mediator involved in cell proliferation, cell cycle regulation, cell survival, angiogenesis, cell migration, etc., and genetic mutations are observed in cancer cells. The inhibitor targeting BRAF may be Dabrafenib, Encorafenib (LGX818), or Vemurafenib.

[0194] As used herein, the term "pan-RAF" encompasses RAF family members such as BRAF, ARAF, and CRAF, and inhibitors targeting pan-RAF may be Naporafenib, Belvarafenib, or Sorafenib. Some pan-RAF inhibitors can broadly inhibit the RAS-RAF-MEK-ERK signaling pathway (e.g., Avutometinib).

[0195] In this specification, the term "SHP2 (Src homology region 2 domain-containing phosphatase-2)", also called Tyrosine-protein phosphatase non-receptor type 11 (PTPN11) or Protein-tyrosine phosphatase 1D / 2C (PTP-1D / 2C), is known as a signaling molecule that regulates various cellular processes including cell growth, differentiation, mitotic cycle, and oncogenic transformation. Activating mutations of SHP2 are found in neuroblastoma, melanoma, acute myeloid leukemia, breast cancer, lung cancer, and colon cancer. Inhibitors targeting SHP2 may include, for example, PF-07284892, RMC-4630 (SHP2-IN-7) or Batoprotafib (TNO155).

[0196] As used herein, the term "SRC (Proto-oncogene tyrosine-protein kinase)" refers to a non-receptor tyrosine kinase protein, also known as c-Src, which regulates embryonic development and cell growth, and its elevated activity level is known to be associated with cancer progression. An SRC inhibitor may be, for example, Dasatinib or Bosutinib.

[0197] As used herein, the term "LCK (lymphocyte-specific protein tyrosine kinase)" belongs to the SFK (Src kinase family) and activates T cell receptor signaling. Mutations and dysfunction of LCK impair T cell activation, and cancer, asthma, diabetes mellitus 1, rheumatoid arthritis, psoriasis, systemic lupus erythematosus, and inflammatory bowel diseases (Crohn's disease and ulcerative colitis) are known to be associated with overexpression of LCK. An inhibitor targeting LCK may be, for example, WH-4-023.

[0198] As used herein, the term "PARP (Poly[ADP-ribose]polymerase)" refers to a protein that recognizes damaged DNA in the nucleus, becomes activated, and then activates DNA repair-related proteins. A PARP target inhibitor inhibits the proliferation of cancer cells by inhibiting DNA repair in cancer cells. A specific example of the PARP target inhibitor may be Olaparib, Talazoparib, Niraparib, or Rucaparib.

[0199] As used herein, the term "DNA methyltransferase (DNMT)" refers to an enzyme that attaches a methyl group to the histone protein that wraps DNA, thereby suppressing gene expression. The DMNT target inhibitor exhibits anticancer activity by inhibiting hypermethylation of tumor suppressor genes and inducing normal expression of the tumor suppressor genes. Specific examples of the DNMT target inhibitor may be Azacitidine, Decitabine, or Guadecitabine.

[0200] As used herein, the term "CDK (Cyclin-dependent kinase) 4 / 6" refers to a protein that promotes cell growth by regulating the cell cycle, and is overactive in the development and progression of various malignant tumors. CDK4 / 6 targeted inhibitors exhibit anticancer activity by inhibiting cell proliferation and inducing apoptosis by inhibiting the cell cycle of cancer cells. The CDK4 / 6 targeted inhibitor may be Abemaciclib (LY2835219), Ribociclib, or Palbociclib.

[0201] As used herein, "Aurora kinase" is a serine / threonine kinase essential for cell proliferation and a phosphotransferase enzyme that helps dividing cells distribute their genetic material to daughter cells. Aurora kinases play a crucial role in cell division by controlling chromatid segregation, and defects in segregation can lead to tumorigenesis. Aurora A (Aurora 2) functions during mitotic prophase and is involved in the proper replication and separation of centrosomes (the microtubule-organizing centers of eukaryotic cells). Aurora B (Aurora 1) is responsible for attaching the mitotic spindle to the centromere. Aurora C (AURKC) acts in germ cells. Aurora A-targeting inhibitors may include MK-5108, Hesperadin, LY3295668, etc. Pan-Aurora-targeting inhibitors may include Danusertib, AMG-900, Reversine, Tozasertib (VX-680), etc.

[0202] In this specification, "WEE1" is a 96 kDa nuclear kinase belonging to the Ser / Thr protein kinase family, also called mitosis inhibitor protein kinase Wee1. Mitosis-promoting factor (MPF) regulates apoptosis induced by DNA damage, and negative regulation of MPF by WEE1 induces abnormal mitosis, resulting in resistance to apoptosis induced by DNA damage. A WEE1-targeting inhibitor can reduce the sensitivity to apoptosis induced by DNA damage in cancer cells by modulating WEE1. WEE1-targeting inhibitors may include MK-1775 (Adavosertib), Azenosertib (ZN-C3), ZNL-02-096, etc.

[0203] In this specification, "PKMYT1 (protein kinase, membrane-associated tyrosine / threonine 1)" belongs to the Wee1 protein kinase family and, as a regulator of CDK1 phosphorylation, is a potent therapeutic target for the treatment of certain types of DNA damage-responsive cancers through synthetic lethality of CCNE1 amplification. PKMYT1-targeting inhibitors may include RP-6306, GSK-1520489A, etc.

[0204] As used herein, "HSP90 (heat shock protein 90)" is a chaperone protein that helps other proteins fold properly, stabilizes proteins from heat stress, and assists in protein degradation. It can exhibit anticancer effects by stabilizing numerous proteins necessary for tumor growth. HSP90 inhibitors may include BIIB021, BIIB028, MPC-3100, PU-H71, Debio093, SNX-5422, AUY922, and the like.

[0205] In this specification, "A3AR (adenosine A3 receptor; ADORA3)" is a G protein-coupled receptor that binds Gi / Gq and is involved in various intracellular signaling pathways and physiological functions, is overexpressed in pathological human cells, and can mediate cell proliferation and apoptosis. A3AR-targeting therapeutic agents may include Reversine, KF-26777, MRS-545, CAY10498, and the like.

[0206] As used herein, "EZH2 (enhancer of zeste homolog 2)" refers to a histone-lysine N-methyltransferase enzyme encoded by the EZH2 gene, which participates in histone methylation and ultimately transcriptional repression. EZH2 is an attractive target for anticancer therapy because it promotes cancer cell division and proliferation, and is found in higher amounts in a wide range of cancers, including breast, prostate, bladder, uterine, and renal cancers, as well as melanoma and lymphoma, compared to healthy cells. EZH2-targeting inhibitors may include DZNep, EPZ005687, EI1, GSK126, UNC1999, Tazemetostat, Sinefungin, etc.

[0207] In this specification, "ARID1A (AT-rich interactive domain-containing protein 1A)" is a member of the SWI / SNF family, has helicase and ATPase activities, and regulates the transcription of specific genes by altering the chromatin structure around the gene. The ARID domain is a DNA binding domain that can specifically bind to AT-rich DNA sequences known to be recognized by the SWI / SNF complex in the beta-globin locus, and the C-terminus of the protein can stimulate glucocorticoid receptor-dependent transcriptional activation. This gene is frequently mutated in gastric cancer, ovarian clear cell carcinoma, and pancreatic cancer. Inhibitors targeting EZH2 / ARID1A may include GSK-343, etc.

[0208] As used herein, "SUMOylation" is a post-translational modification that covalently attaches a small ubiquitin-like modifier (SUMO) polypeptide to a lysine residue of a target protein. The enzymatic pathway of SUMOylation is very similar to that of ubiquitination and involves an activating enzyme, a conjugating enzyme, a ligase, and a deconjugating enzyme. Dysregulation of the SUMOylation pathway has been observed in cancer and neurological diseases, with SUMO enzymes being upregulated in many cancers, and SUMO levels directly correlate with prognosis and disease progression. SUMOylation inhibitors may include Davidiin, CID9549553, 2-D08, etc.

[0209] As used herein, "Chk1 (checkpoint kinase 1; CHEK1)" is a serine / threonine-specific protein kinase that mediates the DNA damage response (DDR) and cell cycle checkpoint responses. Activation of Chk1 leads to the initiation of cell cycle checkpoints, cell cycle arrest, DNA repair, and apoptosis, thereby preventing damaged cells from progressing through the cell cycle. Chk1 is overexpressed in numerous tumors, including breast, colon, liver, gastric, and nasopharyngeal cancers, and the positive correlation between Chk1 expression and tumor grade and disease recurrence suggests that Chk1 may promote tumor growth. Chk1-targeting inhibitors may include SCH-900776, SRA737, V158411, PF-477736, AZD7762, and LY2880070 (Prexasertib).

[0210] As used herein, the term "Ataxia telangiectasia mutated (ATM) and RAD3-related kinase (ATR)" refers to a protein kinase involved in the cellular response to certain forms of DNA damage (e.g., double-strand breaks and replication stress). Normal cells repair damaged DNA using the ATM / ATR signaling pathway, which regulates the cellular response to double-strand DNA breaks and replication stress, referred to as the DNA Damage Response ("DDR"). In contrast, many cancer cells exhibit a high dependence on DNA repair proteins, including ATR, due to defects in ATM in the DNA repair process. ATR-targeting inhibitors may be Berzosertib (VX-970), Gartisertib (VX-803), or Ceralasertib (AZD6738).

[0211] As used herein, "HDAC (histone deacetylase)" is an enzyme that removes an acetyl group from the ε-N-acetyl lysine amino acid of histone and non-histone proteins, and regulates DNA expression by acetylation and deacetylation by making histones wrap DNA more tightly. HDACs include subgroups such as class I, such as HDAC1, HDAC2, and HDAC3, and class IIA, such as HDAC4, HDAC5, and HDAC7. HDAC inhibitors have shown anticancer efficacy in studies on pancreatic cancer, esophageal squamous cell carcinoma (ESCC), multiple myeloma, prostate carcinoma, gastric cancer, leukemia, breast cancer, liver cancer, ovarian cancer, lung cancer, Hodgkin's lymphoma, and neuroblastoma. HDAC target inhibitors may include Panobinostat (LBH589), Entinostat, Mocetinostat, Trichostatin A, CBUD-1001, Abexinostat (PCI-24781, CRA-024781), etc.

[0212] As used herein, "Akt (protein kinase B: PKB)" refers to a set of serine / threonine-specific protein kinases that play a key role in various cellular processes, such as glucose metabolism, apoptosis, cell proliferation, transcription, and cell death, and are associated with tumor cell survival, proliferation, and invasiveness. Akt activation is commonly observed in human cancer and tumor cells, and tumor cells depend on Akt for survival. Akt-targeting inhibitors may include VQD-002, Perifosine, Miltefosine, MK-2206, AZD5363, Ipatasertib, and the like.

[0213] In this specification, "PLK1 (Polo-like kinase 1)", also called serine / threonine-protein kinase 1 or serine / threonine-protein kinase 13 (STPK13), is a 66 kDa enzyme consisting of 603 amino acids. Many colon and lung cancers are caused by mutations in K-RAS and are known to be dependent on PLK1. Silencing PLK1 expression by RNA interference in cell culture can selectively kill K-RAS cells without harming normal cells. PLK1-targeting inhibitors may include Volasertib, Rigosertib, etc.

[0214] In this specification, "BET (Bromodomain and extraterminal domain protein)" is a bromodomain composed of approximately 110 amino acid proteins that recognize acetylated lysine residues, including BRD2, BRD3, BRD4, and BRDT, and transduce signals transmitted by acetylated lysine residues and transform them into various normal or abnormal phenotypes. Bromodomains translate dysregulated cellular acetylomes into disease phenotypes, and BET is a target for cancer and multiple sclerosis. BET target inhibitors may include JQ1, I-BET 151 (GSK1210151A), I-BET 762 (GSK525762), OTX-015, TEN-010, CPI-203, CPI-0610, Olinone, RVX-208, ABBV-744, LY294002, AZD5153, MT-1, MS645, etc.

[0215] As used herein, "IFG (insulin-like growth factor)" refers to a protein with high sequence similarity to insulin, which is involved in communication between cells and the physiological environment, and includes IGF1 / 2, IGF-1R, IGF-2R, etc. IGF-1 stimulates the growth of prostate cancer and breast cancer cells, and IGFs are being found to be involved in diseases such as cancer and diabetes. IGF1 / 2 or IGF-1R targeting inhibitors may include NVP-ADW742, Figitumumab, Mecasermin, rhIGF-1, BI 885578, etc.

[0216] As used herein, "PIK (phosphatidylinositol kinase)" consists of phosphatidylinositol 3-kinase (PI3K) and phosphatidylinositol 4-kinase (PI4K). PI3K phosphorylates phosphoinositides at the 3-hydroxyl group of the inositol ring and is involved in cell signaling, and PI4K acts on phosphatidylinositol (PI) to produce the second messenger inositol-1,4,5-trisphosphate, and abnormalities of these are associated with cancer. PIK target inhibitors may include Duvelisib, Buparlisib, Copanlisib, Dactolisib, Idelalisib, Parsaclisib, Paxalisib, Taselisib, Zandelisib, Inavolisib, and the like.

[0217] As used herein, "CDK9 (cyclin-dependent kinase 9)" is a cell cycle regulator that is associated with P-TEFb. CDK9 is involved in several protein-protein interaction networks that are often implicated in transcriptional deregulation in cancer. CDK9-targeting inhibitors may include, for example, AZD4573, atuveciclib, VIP152, A-1592668, JSH-150, SLS009, AT-7519, Roscovitine, and the like.

[0218] As used herein, "DHFR (dihydrofolate reductase)" is an enzyme that reduces dihydrofolate to tetrahydrofolate using NADPH as an electron donor, and is a component of the multiprotein complex TAK / P-TEFb, an elongation factor for transcription and function by RNA polymerase II, by phosphorylating the C-terminal domain of the largest subunit of RNA polymerase II. DHFR is responsible for the level of intracellular tetrahydrofolate, and inhibition of DHFR can limit cell growth and proliferation, which is a characteristic of cancer and bacterial infection. DHFR target inhibitors may include Methotrexate, Pralatrexate, Pemetrexed, Raltitrexed, Trimetrexate, Nolatrexed, Piritrexim, Talotrexin, and the like.

[0219] As used herein, the term "STING (Stimulator of Interferon Genes)" refers to an in vivo sensor that recognizes DNA fragments released from cancer cells, stimulates interferon genes, and activates immune cells in the body, such as dendritic cells. The STING agonist exhibits immune-enhancing and tumor angiogenesis-inhibiting effects. Examples of STING agonists include CDNs, SB11285, and DMXAA.

[0220] The compounds disclosed herein can be used in combination with other target inhibitors described above to significantly inhibit the expression and activity of target proteins or genes by enhancing the action of the target inhibitors. Specifically, the compounds disclosed herein not only possess their own anticancer efficacy, but can also enhance the anticancer efficacy of the target inhibitors. Therefore, when the compounds disclosed herein are used in combination with the target inhibitors, excellent anticancer efficacy can be achieved.

[0221] As an example, the compounds of the present disclosure may exhibit enhanced tumor suppression activity when combined with a targeted anticancer agent that acts on one or more targets selected from Bcl-2, EGFR, KRAS, MEK, and RAF. For example, the compounds of the present disclosure may be useful for treating cancer, such as treating KRAS mutant cancers, in combination with a KRAS inhibitor. For example, KRAS mutant cancers may include, but are not limited to, KRAS G12C mutations, KRAS G12D mutations, and KRAS G12V mutations. For example, KRAS mutant cancers may include, but are not limited to, non-small cell lung cancer, colorectal cancer, ovarian cancer, glioblastoma, pancreatic cancer, biliary tract cancer, endometrial cancer, gastric cancer, breast cancer, melanoma, and the like. For example, KRAS G12C inhibitors that exhibit enhanced tumor suppression activity when combined with the compounds of the present disclosure include, but are not limited to, sotorasib, adagrasib, and dibarasib.

[0222] As used herein, the term "oncolytic virus therapy" refers to a therapeutic agent that kills cancer by inserting a specific gene targeting cancer cells into a viable and infectious virus. The oncolytic virus therapy may be Talimogene Laherparepvec.

[0223] As used herein, the term "antibody therapeutic agent" refers to a therapeutic agent that exhibits an anticancer effect by utilizing an antibody that recognizes a specific protein of a cancer cell as an antigen. Antibody therapeutic agents may be Cetuximab, Trastuzumab, Emtansine, Emtansine, Rituximab, Ibritumomab, Tositumomab, Brentuximab, Ofatumumab, Obinutuzumab, Necitumumab, Bevacizumab, Ramucirumab, Nivolumab, Pembrolizumab, Atezolizumab, Durvalumab, Ipilimumab, etc.

[0224] As used herein, the term "immunotherapy" refers to a treatment that uses immune cells such as dendritic cells, natural killer cells, and T cells to activate the immune response in the body and exhibit an anticancer effect. Immunotherapy is used by extracting immune cells from the body, strengthening them, or genetically modifying them, and then reinjecting them into the body. Representative immunotherapy agents include T cell receptor-modified T cells (TCR-T) and chimeric antigen receptor-modified T cells (CAR-T). Specifically, it may be, but is not limited to, Tisagenlecleucel or Axicabtagene Ciloleucel.

[0225] As used herein, the term "immune checkpoint inhibitor" refers to a substance that inhibits the activity of immune checkpoint proteins that suppress the differentiation, proliferation, and activity of immune cells, and is known to eliminate cancer cells by preventing cancer cells from exercising their function of evading the immune system. The immune checkpoint inhibitor may be any one selected from the group consisting of anti-CTLA-4 antibodies, anti-PD-1 antibodies, anti-PD-L1 antibodies, anti-PD-L2 antibodies, anti-B7-H4 antibodies, anti-HVEM antibodies, anti-TIM3 antibodies, anti-GAL9 antibodies, anti-LAG3 antibodies, anti-VISTA antibodies, anti-KIR antibodies, anti-BTLA antibodies, and anti-TIGIT antibodies. In a specific example, the immune checkpoint inhibitor may be, but is not limited to, Ipilimumab, Pembrolizumab, Nivolumab, Cemiplimab, Atezolizumab, Avelumab, and Durvalumab.

[0226] As used herein, the term "ADC (Antibody Drug Conjugate)" refers to a therapeutic agent that exhibits high anticancer efficacy through targeted delivery by chemically combining an antibody and a cytotoxic drug. Examples thereof include Gemtuzumab-Ozogamicin, Brentuximab-Vedotin, Trastuzumab-Emtansine, Inotuzumab-Ozogamicin, and Eribulin-Mesylate.

[0227] The anticancer agent may include one or more anticancer agents. Specifically, the compound, solvate, stereoisomer, or pharmaceutically acceptable salt thereof may be compatible with two anticancer agents. For example, the anticancer agent and a targeted anticancer agent; the anticancer agent and an anticancer virus; the anticancer agent and an antibody therapeutic agent; the anticancer agent and a cell therapeutic agent; and the anticancer agent and an immune checkpoint inhibitor. Furthermore, the anticancer agent and a targeted anticancer agent; the anticancer agent and an antibody therapeutic agent; the anticancer agent and an immunocytotherapy agent; and the anticancer agent and an immune checkpoint inhibitor. Furthermore, the anticancer agent and an antibody therapeutic agent; the anticancer agent and an immune checkpoint inhibitor; and the anticancer agent and an antibody therapeutic agent; and the anticancer agent and an immune checkpoint inhibitor. Furthermore, the anticancer agent and an antibody therapeutic agent; and an antibody therapeutic agent and an immune checkpoint inhibitor.

[0228] The above compound, solvate, stereoisomer, or pharmaceutically acceptable salt thereof may be used in combination with three anticancer agents. A different anticancer agent may be additionally included in addition to the above two anticancer agents.

[0229] The above compounds, solvates, stereoisomers, or pharmaceutically acceptable salts thereof may be used in combination with four anticancer agents. A different anticancer agent may be additionally included in addition to the above three anticancer agents.

[0230] The above compounds, solvates, stereoisomers, or pharmaceutically acceptable salts thereof may be used in combination with five anticancer agents. Additional anticancer agents may be added to the above four anticancer agents.

[0231] The above compounds, solvates, stereoisomers or pharmaceutically acceptable salts thereof can be used in combination with six anticancer agents.

[0232] The above compounds, solvates, stereoisomers or pharmaceutically acceptable salts thereof can be used in combination with anticancer vaccines.

[0233] As used herein, the term "anticancer vaccine" refers to an active immunotherapy method that enhances the body's immune function and eliminates cancer cells by administering tumor-specific antigens (TSA) produced by cancer cells to cancer patients, thereby activating the immune system. Depending on the type of antigen and delivery method, anticancer vaccines include DNA vaccines, peptide vaccines, and cell vaccines. Currently, cell vaccines and DNA vaccines, which introduce antigens, are the most representative vaccines being developed.

[0234] The above compounds, solvates, stereoisomers, or pharmaceutically acceptable salts thereof may be used in combination with the above anticancer agents and anticancer vaccines. Here, the compounds and anticancer agents are the same as those described above.

[0235] Another aspect provides a method of preventing or treating an SOS1-mediated disease comprising administering to a subject a compound of the present invention comprising a compound of formula I, a solvate, stereoisomer or pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same.

[0236] Any terms or elements mentioned in the description of the above method that are identical to those mentioned above are as described above.

[0237] The above administration may be oral or parenteral. In case of oral administration, the active ingredient may be administered in an amount of 0.01 to 1000 mg, more specifically 0.1 to 300 mg per kg of body weight per day, and in case of parenteral administration, the active ingredient may be administered in an amount of 0.01 to 100 mg, more specifically 0.1 to 50 mg per kg of body weight per day, and may be administered in one or more divided doses. The dosage for a specific individual or patient should be determined in light of various related factors such as the patient's weight, age, sex, health condition, diet, administration time, administration method, and severity of the disease, and may be appropriately increased or decreased by a specialist.

[0238] As used herein, the term "subject" means a subject in need of treatment for a disease, and more specifically, a mammal such as a human or non-human primate, mouse, dog, cat, horse, and cow.

[0239]

[0240] Another aspect provides a method of inhibiting the interaction of SOS1 and a RAS family protein, and / or SOS1 and RAC1, in a sample or cell, comprising administering to the sample or cell a compound of the present invention comprising a compound of formula I, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof.

[0241] Another aspect provides a pharmaceutical use of a compound of the present invention comprising a compound of formula I, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof, for the prevention or treatment of a SOS1-mediated disease; or a use of a compound of the present invention comprising a compound of formula I, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for the prevention or treatment of a SOS1-mediated disease.

[0242] Any terms or elements mentioned in the description of the above method or use that are identical to those already mentioned are as described above.

[0243] The compounds of the present invention, including compounds of formula I, solvates, stereoisomers, or pharmaceutically acceptable salts thereof, exhibit effective inhibitory activity against SOS1. Specifically, they are useful for the prevention or treatment of SOS1-mediated diseases, specifically diseases associated with abnormal activity of SOS1 and / or RAS family proteins, by inhibiting the interaction between SOS1 and RAC1. In addition, the compounds of the present invention exhibit excellent efficacy in that they exhibit optimal DMPK properties and good PK exposure, and the risk of drug interactions is reduced.

[0244] Figure 1 shows the results of confirming the CYP3A4 induction ability of the compounds of Examples 1 and 56 in human hepatocytes.

[0245] Figure 2 shows the results confirming the enhanced tumor suppression effect according to the combined administration of compounds of Examples 1 and 56 and sotorasib in the H358 tumor subcutaneous transplantation model. **** indicates P < 0.0001 or less.

[0246] Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following examples.

[0247] Manufacturing Example 1: (1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethanamine (A-1)

[0248]

[0249] Step 1: Synthesis of 1-bromo-3-(1,1-difluoroethyl)-2-fluorobenzene

[0250] A solution of 1-(3-bromo-2-fluorophenyl)ethan-1-one (4 g, 18.43 mmol) and DAST (23.77 g, 147.44 mmol, 19.48 mL) in DCM (60 mL) was degassed and purged three times with N2, and stirred at 50 °C for 60 h under N2 atmosphere. The reaction mixture was slowly quenched with ice-water (40 mL) and basified (pH = 7-8) with saturated NaHCO3. The aqueous layer was extracted with DCM (50 mL Х 3), and the organic layer was washed with brine (20 mL Х 3). The organic layer was dehydrated over anhydrous Na2SO4 and filtered under reduced pressure to obtain the crude product. This was purified by silica gel column chromatography (0% EA in petroleum ether) to give 1-bromo-3-(1,1-difluoroethyl)-2-fluorobenzene (3.5 g, 79.45% yield) as a colorless oil. 1 H NMR (400 MHz, DMSO-d6) δ ppm 7.89 (br t,J= 7.2 Hz, 1H), 7.60 (br t,J= 7.2 Hz, 1H), 7.29 (t,J= 8.0 Hz, 1H), 2.02 (t,J= 19.2 Hz, 3H)

[0251] Step 2: Synthesis of 1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethanone

[0252] A mixture of 1-bromo-3-(1,1-difluoroethyl)-2-fluorobenzene (3.5 g, 14.64 mmol), 1-tributyl(1-ethoxyvinyl)stannane (5.66 g, 15.67 mmol, 5.29 mL), TEA (2.96 g, 29.28 mmol, 4.08 mL) and Pd(PPh3)2Cl2 (1.03 g, 1.46 mmol) in dioxane (40 mL) was degassed, purged three times with N2, and stirred at 80 °C for 16 h under N2 atmosphere. HCl solution (4 N, 60 mL) was added to the mixture, and stirred at 25 °C for 1 h. Water (30 mL) was added to the reaction mixture, and extracted with EtOAc (50 mL Х 3). The combined organic layers were washed with brine (50 mL X 3), dehydrated over anhydrous Na2SO4, and concentrated under reduced pressure to obtain a residue. This was purified by silica gel column chromatography (3% EtOAc in petroleum ether) to give 1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethanone (2.5 g, 84.45% yield) as a colorless oil. 1 H NMR (400 MHz, DMSO-d6) δ ppm 7.95 (br t,J= 7.2 Hz, 1H), 7.82 (br t,J= 7.2 Hz, 1H), 7.44 (t,J= 7.6 Hz, 1H), 2.62 (d,J= 4.0 Hz, 3H), 2.05 (t,J= 19.2 Hz, 3H).

[0253] Step 3: Synthesis of (NE,S)-N-[1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethylidene]-2-methylpropane-2-sulfinamide

[0254] A mixture of 1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethanone (2.7 g, 13.35 mmol), (S)-2-methylpropane-2-sulfinamide (2.43 g, 20.03 mmol), and Ti(OEt)4 (9.14 g, 40.06 mmol, 8.31 mL) in THF (40 mL) was degassed, purged three times with N2, and stirred at 80 °C for 16 h under N2 atmosphere. The mixture was poured into water (50 mL), filtered, and extracted with EtOAc (60 mL Х 3). The organic layer was washed with brine (40 mL Х 2), dehydrated over anhydrous Na2SO4, and concentrated under reduced pressure to obtain the crude product. This was purified by silica gel column chromatography (5% EtOAc in petroleum ether) to give (NE,S)-N-[1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethylidene]-2-methylpropane-2-sulfinamide (3.4 g, 76.29% yield, 91.5% purity) as a yellow oil. MS (ESI) m / z = 306.2 [M+H] + .

[0255] Step 4: Synthesis of (S)-N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethyl]-2-methylpropane-2-sulfinamide

[0256] To a solution of (NE,S)-N-[1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethylidene]-2-methylpropane-2-sulfinamide (3.4 g, 11.13 mmol) in THF (50 mL) was added L-selectride (1 M, 27.84 mL) at -70 °C, degassed and purged with N2 three times, and the mixture was stirred at -70 °C for 3 h under N2 atmosphere. The reaction mixture was quenched by slowly adding saturated NH4Cl aqueous solution (80 mL) dropwise at -70 °C, diluted with EtOAc (50 mL), and extracted with EtOAc (40 mL Х 5). The organic layer was dried over anhydrous Na2SO4 and concentrated under reduced pressure to obtain the crude product. This was purified by silica gel column chromatography (25% EtOAc in petroleum ether) to give the major product (S)-N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethyl]-2-methylpropane-2-sulfinamide (2.3 g, 65.86% yield, 98% purity, 99.3% enantiomeric excess) as a light yellow oil. 1 H NMR (400 MHz, DMSO-d6) δ ppm ppm 7.63 (t,J= 6.8 Hz, 1H), 7.51 - 7.40 (m, 1H), 7.33 - 7.26 (m, 1H), 5.54 (d,J= 5.6 Hz, 1H), 4.72 (quint,J= 6.8 Hz, 1H), 2.00 (t,J= 19.2 Hz, 3H), 1.49 (d,J= 6.8 Hz, 3H), 1.09 (s, 9H); MS (ESI) m / z = 308.0 [M+H] + .

[0257] Step 5: Synthesis of (1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethanamine

[0258] A solution of (S)-N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethyl]-2-methylpropane-2-sulfinamide (500 mg, 1.63 mmol) in dioxane (2 mL) and HCl / dioxane (4 N, 25 mL) was degassed and purged three times with N2, and the mixture was stirred at 25 °C for 0.5 h under N2 atmosphere. The mixture was concentrated under reduced pressure to give (1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethanamine (A-1,350 mg, 89.78% yield, HCl) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.75 (br s, 3H), 7.88 (t,J= 7.2 Hz, 1H), 7.59 (t,J= 7.2 Hz, 1H), 7.41 (t,J= 7.6 Hz, 1H), 4.66 (br d,J= 5.2 Hz, 1H), 2.02 (t,J= 19.2 Hz, 3H), 1.56 - 1.53 (m, 3H); MS (ESI) m / z = 204.1 [M+H] + .

[0259] Manufacturing Example 2: (1R)-1-[2-fluoro-3-(1,1,2-trifluoroethyl)phenyl]ethanamine (A-2)

[0260]

[0261] Step 1: Synthesis of 1-(2-fluoro-3-iodophenyl)ethanol

[0262] To a solution of 2-fluoro-3-iodo-benzaldehyde (4.0 g, 16.00 mmol) in THF (40 mL) was added MeMgBr (3 M, 8.00 mL) dropwise at -78 °C and stirred for 3 h. The reaction mixture was poured into a saturated aqueous NH4Cl solution (50 mL) and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na2SO4, and concentrated under reduced pressure to obtain the crude product. Purification by silica gel column chromatography (7% EtOAc in petroleum ether) gave 1-(2-fluoro-3-iodophenyl)ethanol (4.45 g, 83.63% yield) as a yellow oil. 1 H NMR (400 MHz, chloroform-d) δ ppm 7.72-7.64 (m, 1H), 7.55-7.45 (m, 1H), 6.93 (t,J= 7.6 Hz, 1H), 5.20 (q,J= 6.4 Hz, 1H), 1.52 (d,J= 6.4 Hz, 3H).

[0263] Step 2: Synthesis of 1-(2-fluoro-3-iodophenyl)ethanone

[0264] To a solution of 1-(2-fluoro-3-iodophenyl)ethanol (4.45 g, 16.73 mmol) in MeCN (50 mL) were added TPAP (Tetrapropylammonium perruthenate; 587.80 mg, 1.67 mmol) and NMO (N-methyl morpholine-N-oxide; 2.94 g, 25.09 mmol), and the mixture was stirred at 20 °C for 2 h. The mixture was filtered and concentrated under reduced pressure to obtain the crude product, which was then purified by silica gel column chromatography (0% EtOAc in petroleum ether) to give 1-(2-fluoro-3-iodophenyl)ethanone (3.8 g, 12.95 mmol, 77.44% yield, 90% purity) as a white solid. 1H NMR (400 MHz, chloroform-d) δ ppm 7.97-7.88 (m, 1H), 7.85-7.78 (dm, 1H), 7.00 (t,J= 7.6 Hz, 1H), 2.65 (d,J= 5.2 Hz, 3H).

[0265] Step 3: Synthesis of ethyl 2-(3-acetyl-2-fluorophenyl)-2,2-difluoroacetate

[0266] To a solution of 1-(2-fluoro-3-iodophenyl)ethanone (3.0 g, 11.36 mmol) and ethyl 2-bromo-2,2-difluoroacetate (6.92 g, 34.09 mmol, 4.38 mL) in DMSO (30 mL) was added Cu (2.17 g, 34.09 mmol), and the mixture was stirred at 80 °C for 12 h. The reaction mixture was poured into water (50 mL) and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na2SO4, and concentrated under reduced pressure to obtain the crude product. Purification by silica gel column chromatography (5% EtOAc in petroleum ether) afforded ethyl 2-(3-acetyl-2-fluorophenyl)-2,2-difluoroacetate (1.8 g, 56.05% yield, 92.07% purity) as a colorless oil. 1 H NMR (400 MHz, chloroform-d) δ ppm 8.08-8.00 (m, 1H), 7.87-7.81 (m, 1H), 7.36 (t,J= 7.6 Hz, 1H), 4.42-4.37 (m, 2H), 2.66 (d,J= 5.2 Hz, 3H), 1.35 (t,J= 7.2 Hz, 3H); MS (ESI) m / z = 261.0 [M+H] + .

[0267] Step 4: Synthesis of ethyl (R,E)-2-(3-(1-((tert-butylsulfinyl)imino)ethyl)-2-fluorophenyl)-2,2-difluoroacetate

[0268] To a solution of ethyl 2-(3-acetyl-2-fluorophenyl)-2,2-difluoroacetate (1.8 g, 6.92 mmol) and (R)-2-methylpropane-2-sulfinamide (1.26 g, 10.38 mmol) in THF (20 mL) was added Ti(OEt)4 (4.73 g, 20.75 mmol), and the mixture was stirred at 80 °C for 16 h. The reaction mixture was poured into water (30 mL) and EtOAc (30 mL), filtered, and the filtrate was extracted with EtOAc. The combined organic layers were washed with brine, dried over Na2SO4, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (6% EtOAc in petroleum ether) to give ethyl (R,E)-2-(3-(1-((tert-butylsulfinyl)imino)ethyl)-2-fluorophenyl)-2,2-difluoroacetate (1.9 g, 74.09% yield, 98.03% purity) as a yellow oil. 1 H NMR (400 MHz, chloroform-d) δ ppm 7.86-7.72 (m, 2H), 7.32 (t, J= 7.6 Hz, 1H), 4.44-4.30 (m, 2H), 2.77 (s, 3H), 1.32 (s, 9H); MS (ESI) m / z = 364.0 [M+H] + .

[0269] Step 5: Synthesis of (R)-N-[(1R)-1-[3-(1,1-difluoro-2-hydroxyethyl)-2-fluorophenyl]ethyl]-2-methylpropane-2-sulfinamide

[0270] To a solution of ethyl (R,E)-2-(3-(1-((tert-butylsulfinyl)imino)ethyl)-2-fluorophenyl)-2,2-difluoroacetate (900 mg, 2.48 mmol) in THF (10 mL) and H2O (0.2 mL) was added NaBH4 (210 mg, 5.55 mmol) at -78 °C, slowly warmed to 10 °C, and stirred at 10 °C for 2 h. The reaction mixture was poured into iced water (30 mL) and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na2SO4, and concentrated under reduced pressure to obtain the crude product. After primary purification by silica gel column chromatography (50% EtOAc in petroleum ether), two diastereoisomers (approximately 3:1 ratio) were separated and purified using preparative HPLC (Xtimate C18 150*40 mm*10 um, mobile phase: [water (NH3H2O)-MeCN]; B%: 25%-55%, 10 min). MeCN was removed under reduced pressure, and the remaining solvent was removed by lyophilization to obtain the major product (R)-N-[(1R)-1-[3-(1,1-difluoro-2-hydroxyethyl)-2-fluorophenyl]ethyl]-2-methylpropane-2-sulfinamide (440 mg, 50.90% yield, 92.65% purity, enantiomeric excess >99%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ ppm 7.70 (t,J= 6.8 Hz, 1H), 7.48-7.39 (m, 1H), 7.35-7.25 (m, 1H), 5.87 (d,J= 7.6 Hz, 1H), 5.70 (t,J= 6.4 Hz, 1H), 4.68 (quint,J= 7.2 Hz, 1H), 3.90 (dt,J= 6.4, 14.4 Hz, 2H), 1.40 (d,J= 6.8 Hz, 3H), 1.10 (s, 9H); MS (ESI) m / z = 324.3 [M+H] + .

[0271] Step 6: Synthesis of (R)-N-[(1R)-1-[2-fluoro-3-(1,1,2-trifluoroethyl)phenyl]ethyl]-2-methylpropane-2-sulfinamide

[0272] To a solution of (R)-N-[(1R)-1-[3-(1,1-difluoro-2-hydroxyethyl)-2-fluorophenyl]ethyl]-2-methylpropane-2-sulfinamide (4 g, 12.37 mmol) and N,N-diethylethanamine trihydrofluoride (5.98 g, 37.11 mmol, 6.05 mL) and TEA (7.51 g, 74.22 mmol, 10.33 mL) in anhydrous THF (60 mL) was added 1,1,2,2,3,3,4,4,4-nonafluorobutane-1-sulfonyl fluoride (PBSF) (11.21 g, 37.11 mmol, 6.53 mL) dropwise at 20 °C and stirred at 50 °C for 24 h. The crude product was obtained by concentration under reduced pressure. Purification by silica gel column chromatography (38% EtOAc in PE) afforded the major product (R)-N-[(1R)-1-[2-fluoro-3-(1,1,2-trifluoroethyl)phenyl]ethyl]-2-methylpropane-2-sulfinamide (4.4 g, 87.46% yield, 80% purity) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ ppm 7.78 (t,J= 7.2 Hz, 1H), 7.55-7.46 (m, 1H), 7.40-7.33 (m, 1H), 5.88 (d,J= 8.0 Hz, 1H), 5.07-4.85 (m, 2H), 4.70 (quin,J= 7.2 Hz, 1H), 1.42 (d,J= 6.8 Hz, 3H), 1.10 (s, 9H); MS (ESI) m / z = 326.2 [M+H] + .

[0273] Step 7: Synthesis of (1R)-1-[2-fluoro-3-(1,1,2-trifluoroethyl)phenyl]ethanamine

[0274] To a solution of (R)-N-[(1R)-1-[2-fluoro-3-(1,1,2-trifluoroethyl)phenyl]ethyl]-2-methylpropane-2-sulfinamide (4.4 g, 13.52 mmol) in dioxane (4 mL) at 0 °C was added 4N HCl / dioxane (20 mL) and stirred at 25 °C for 1 h. The mixture was concentrated under reduced pressure to give (1R)-1-[2-fluoro-3-(1,1,2-trifluoroethyl)phenyl]ethanamine (A-2, 3.8 g, 95.28% yield, 75% purity, HCl salt) as a yellow oil. 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.80 (br s, 3H), 7.97 (t,J= 7.2 Hz, 1H), 7.64 (t,J= 6.8 Hz, 1H), 7.51-7.41 (m, 1H), 5.13-4.88 (m, 2H), 4.71-4.58 (m, 1H), 1.55 (d, J = 6.8 Hz, 3H); MS (ESI) m / z = 222.0 [M+H] + .

[0275] Manufacturing Example 2-1: (1R)-1-[2-methyl-3-(trifluoromethyl)phenyl]ethanamine (A-3)

[0276]

[0277] Step 1: Synthesis of (NE, S)-2-methyl-N-[1-[2-methyl-3-(trifluoromethyl)phenyl]ethylidene]propane-2-sulfinamide

[0278] (NE, S)-2-methyl-N-[1-[2-methyl-3-(trifluoromethyl)phenyl]ethylidene]propane-2-sulfinamide (2.3 g, 50.8% yield) was obtained as a yellow oil using the same method as in step 3 of Preparation Example 1 using commercially available 1-[2-methyl-3-(trifluoromethyl)phenyl]ethanone. 1H NMR (400 MHz, CDCl3) δ ppm 7.63 - 7.74 (m, 1 H), 7.30 - 7.45 (m, 2 H), 2.71 (s, 2 H), 2.48 (br s, 3 H), 1.62 (s, 1 H), 1.25 - 1.34 (m, 9 H); MS (ESI) m / z =306.0 [M+H] + .

[0279] Step 2: Synthesis of (S)-2-methyl-N-[(1R)-1-[2-methyl-3-(trifluoromethyl)phenyl]ethyl]propane-2-sulfinamide

[0280] (S)-2-methyl-N-[(1R)-1-[2-methyl-3-(trifluoromethyl)phenyl]ethylidene]propane-2-sulfinamide was used in the same manner as in step 4 of Preparation Example 1 to obtain (S)-2-methyl-N-[(1R)-1-[2-methyl-3-(trifluoromethyl)phenyl]ethyl]propane-2-sulfinamide (1 g, 42.9% yield, 99.4% purity) as a white oil. 1 H NMR (400 MHz, DMSO-d6) δ ppm 7.72 (d,J= 8.0 Hz, 1H), 7.55 (d,J= 8.0 Hz, 1H), 7.44 - 7.32 (m, 1H), 5.44 (d,J= 5.6 Hz, 1H), 4.75 (quin,J= 6.4 Hz, 1H), 2.40 (s, 3H), 1.45 (d,J= 6.8 Hz, 3H), 1.09 (s, 9H); MS (ESI) m / z =307.9 [M+H] + .

[0281] Step 3: Synthesis of (1R)-1-[2-methyl-3-(trifluoromethyl)phenyl]ethanamine

[0282] (S)-2-methyl-N-[(1R)-1-[2-methyl-3-(trifluoromethyl)phenyl]ethyl]propane-2-sulfinamide was used in the same manner as in step 5 of Preparation Example 1 to obtain (1R)-1-[2-methyl-3-(trifluoromethyl)phenyl]ethanamine (A-3, 1.08 g, crude product, HCl salt) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ ppm 8.52 (br s, 3H), 7.90 (d, J = 8.0 Hz, 1H), 7.71 (d, J = 8.0 Hz, 1H), 7.40 - 7.58 (m, 1H), 4.63 - 4.89 (m, 1H), 2.44 (s, 3H), 1.50 (d, J = 6.4 Hz, 3H); MS (ESI) m / z = 244.9 [M+H+MeCN] + .

[0283] Manufacturing Example 2-2: (1R)-1-[2-fluoro-3-(trifluoromethyl)phenyl]ethanamine (A-4)

[0284]

[0285] Step 1: Synthesis of (NE,S)-N-[1-[2-fluoro-3-(trifluoromethyl)phenyl]ethylidene]-2-methyl-propane-2-sulfinamide

[0286] Using commercially available 1-[2-fluoro-3-(trifluoromethyl)phenyl]ethanone, (NE,S)-N-[1-[2-fluoro-3-(trifluoromethyl)phenyl]ethylidene]-2-methyl-propane-2-sulfinamide (2.08 g, 62.57% yield, 85.77% purity) was obtained as a yellow oil in the same manner as in step 3 of Preparation Example 1. MS (ESI) m / z = 310.1 [M+H] + .

[0287] Step 2: Synthesis of (S)-N-[(1R)-1-[2-fluoro-3-(trifluoromethyl)phenyl]ethyl]-2-methyl-propane-2-sulfinamide

[0288] (NE,S)-N-[1-[2-fluoro-3-(trifluoromethyl)phenyl]ethylidene]-2-methyl-propane-2-sulfinamide was prepared in the same manner as in step 4 of Preparation Example 1, to give (S)-N-[(1R)-1-[2-fluoro-3-(trifluoromethyl)phenyl]ethyl]-2-methyl-propane-2-sulfinamide (1.23 g, 56.49% yield, 92.35% purity, 98.28% ee) as a colorless oil. MS (ESI) m / z = 312.1 [M+H] + .

[0289] Step 3: Synthesis of (1R)-1-[2-fluoro-3-(trifluoromethyl)phenyl]ethanamine

[0290] (S)-N-[(1R)-1-[2-fluoro-3-(trifluoromethyl)phenyl]ethyl]-2-methyl-propane-2-sulfinamide was used in the same manner as in step 5 of Preparation Example 1 to obtain (1R)-1-[2-fluoro-3-(trifluoromethyl)phenyl]ethanamine (A-4,850 mg, 96.04% yield, 97.25% purity, HCl) as a white solid. MS (ESI) m / z = 208.0 [M+H] + .

[0291] Manufacturing Example 2-3: (1R)-1-[2-trideuterio-3-(trifluoromethyl)phenyl]ethanamine (A-5)

[0292]

[0293] Step 1: Synthesis of 2-(trideuterio)-3-(trifluoromethyl)benzoic acid

[0294] A solution of 2,2,6,6-tetramethylpiperidine (70.58 g, 499.69 mmol, 84.83 mL) in THF (250 mL) was degassed, filled with nitrogen gas, cooled to 0 °C, and n-BuLi (1.6 M in hexane, 312.30 mL) was slowly added, and the mixture was stirred for 1 h. Then, 3-(trifluoromethyl)benzoic acid (19 g, 99.94 mmol) in THF (100 mL) was slowly added to the reaction mixture, and the mixture was stirred at 0 °C for 4 h under nitrogen conditions. The reaction mixture was cooled to -78 °C, and trideuterioiodomethane (57.95 g, 399.75 mmol, 24.88 mL) in THF (50 mL) was slowly added, and the mixture was stirred at 25 °C for 1 h. The reaction mixture was quenched with water (100 mL), acidified with 6 M HCl (100 mL) solution, and extracted with EtOAc (200 mLХ3). The combined organic layers were dried over Na2SO4 and concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (10% MeOH in DCM) to obtain the crude product.

[0295] This was purified by preparative HPLC (Column: YMC Triart C18 250*50mm*7um; Mobile phase: [water (0.05%HCl) -MeCN]; Gradient: 20%-68% B for 22 min). MeCN was removed by concentration under reduced pressure, and the residual solvent was removed by lyophilization to obtain 2.1 g, 63.4% pure crude product. This was purified by preparative HPLC (Column: Welch Xtimate C18 250*50mm*10um; Mobile phase: [water (0.225% FA) -MeCN]; Gradient: 0%-30% B for 20 min). MeCN was removed by concentration under reduced pressure, and the residual solvent was removed by lyophilization to obtain 2-(trideuterio)-3-(trifluoromethyl)benzoic acid (500 mg, 2.5% yield, 95.5% purity) as a yellow oil. 1H NMR (400 MHz, DMSO-d6) δ ppm 7.61 (d,J= 7.6 Hz, 1H), 7.57 (d,J= 7.6 Hz, 1H), 7.33 - 7.27 (m, 1H); MS (ESI) m / z = 208.2 [M+H] + .

[0296] Step 2: Synthesis of N-methoxy-N-methyl-2-(trideuterio)-3-(trifluoromethyl)benzamide

[0297] A solution of 2-(trideuterio)-3-(trifluoromethyl)benzoic acid (1.08 g, 5.21 mmol), N,O-dimethylhydroxylamine (559.37 mg, 5.73 mmol), HATU (2.97 g, 7.82 mmol), and DIEA (2.02 g, 15.64 mmol, 2.72 mL) in DMF (8 mL) was stirred at 25 °C for 3 h. The reaction mixture was poured into water (30 mL) and extracted with EtOAc (30 mL Х 3). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure to give the crude product. Purification by silica gel column chromatography (2% MeOH in DCM) gave N-methoxy-N-methyl-2-(trideuterio)-3-(trifluoromethyl)benzamide (1.19 g, 80.82% yield, 88.6% purity) as a yellow oil. 1 H NMR (400 MHz, DMSO-d6) δ ppm 7.75 (br d,J= 7.6 Hz, 1H), 7.59 (d,J= 7.6 Hz, 1H), 7.52 - 7.40 (m, 1H), 2.69 (s, 6H); MS (ESI) m / z = 251.1 [M+H] + .

[0298] Step 3: Synthesis of 1-(2-(trideuterio)-3-(trifluoromethyl)phenyl)ethanone

[0299] To a solution of N-methoxy-N-methyl-2-(trideuterio)-3-(trifluoromethyl)benzamide (1.19 g, 4.76 mmol) in THF (20 mL) was slowly added MeMgBr (3 M, 4.76 mL) at -78 °C and stirred for 3 h. The reaction mixture was poured into a saturated aqueous solution of NH4Cl and extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, and concentrated under reduced pressure to give the crude product. Purification by silica gel column chromatography (2% MeOH in DCM) gave 1-(2-(trideuterio)-3-(trifluoromethyl)phenyl)ethanone (880 mg, 80.05% yield, 88.77% purity) as a yellow oil. 1 H NMR (400 MHz, DMSO-d6) δ ppm 7.93 (d, J= 8.0 Hz, 1H), 7.82 (d, J= 8.0 Hz, 1H), 7.56 - 7.46 (m, 1H), 2.58 (s, 3H); MS (ESI) m / z = 205.9 [M+H] +

[0300] Step 4: Synthesis of (NE,S)-N-[1-[2-(trideuterio)-3-(trifluoromethyl)phenyl]ethylidene]-2-methyl-propane-2-sulfinamide

[0301] Using 1-(2-(trideuterio)-3-(trifluoromethyl)phenyl)ethanone, (NE,S)-N-[1-[2-(trideuterio)-3-(trifluoromethyl)phenyl]ethylidene]-2-methyl-propane-2-sulfinamide (928 mg, 66.41% yield, 94.64% purity) was obtained as a yellow oil in the same manner as in step 3 of Preparation Example 1. 1 H NMR (400 MHz, DMSO-d6) δ ppm 7.75 (br d,J= 7.6 Hz, 1H), 7.62 (br d,J= 7.6 Hz, 1H), 7.51 - 7.47 (m, 1H), 2.64 (s, 3H), 1.19 (s, 9H); MS (ESI) m / z = 309.0 [M+H] + .

[0302] Step 5: Synthesis of (S)-N-[(1R)-1-[2-(trideuterio)-3-(trifluoromethyl)phenyl]ethyl]-2-methyl-propane2-sulfinamide

[0303] (NE,S)-N-[1-[2-(trideuterio)-3-(trifluoromethyl)phenyl]ethylidene]-2-methyl-propane-2-sulfinamide was prepared in the same manner as in step 4 of Preparation Example 1, to give (S)-N-[(1R)-1-[2-(trideuterio)-3-(trifluoromethyl)phenyl]ethyl]-2-methyl-propane2-sulfinamide (700 mg, 58.81% yield, 77.8% purity) as a white solid. MS (ESI) m / z = 311.4 [M+H] + .

[0304] Step 6: Synthesis of (1R)-1-[2-trideuterio-3-(trifluoromethyl)phenyl]ethanamine

[0305] (S)-N-[(1R)-1-[2-(trideuterio)-3-(trifluoromethyl)phenyl]ethyl]-2-methyl-propane2-sulfinamide was used in the same manner as in step 5 of Preparation Example 1 to obtain (1R)-1-[2-trideuterio-3-(trifluoromethyl)phenyl]ethanamine (A-5, 115 mg, 98.06% yield, HCl salt) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.49 (br s, 3H), 7.88 (d,J= 7.6 Hz, 1H), 7.71 (d,J= 7.6 Hz, 1H), 7.57 - 7.47 (m, 1H), 4.78 - 4.68 (m, 1H), 1.50 (d,J=6.8 Hz, 3H); MS (ESI) m / z = 206.9 [M+H] + .

[0306] Manufacturing Example 3: Tributyl-[3-(trideuteriomethyl)triazol-4-yl]stanane (T-1)

[0307]

[0308] Step 1: Synthesis of 1-(trideuteriomethyl)triazole

[0309] To a solution of 1H-triazole (1.99 g, 28.74 mmol, 1.67 mL) in THF (25 mL) were added K2CO3 (7.95 g, 57.49 mmol) and trideuterio(iodo)methane (5 g, 34.49 mmol, 2.15 mL), and the mixture was stirred at 25 °C for 12 h. The reaction mixture was poured into water (20 mL) and extracted with EtOAc (30 mL Х 3). The combined organic layers were washed with brine (30 mL Х 2), dried over Na2SO4, and concentrated under reduced pressure to afford the crude product 1-(trideuteriomethyl)triazole (1.18 g, 47.56% yield) as a yellow oil. 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.06 (s, 1H), 7.70 (s, 1H).

[0310] Step 2: Synthesis of tributyl-[3-(trideuteriomethyl)triazol-4-yl]stannane

[0311] A solution of 1-(trideuteriomethyl)triazole (1 g, 11.61 mmol) in THF (20 mL) was cooled to -78 °C, n-BuLi (2.5 M, 5.20 mL) was slowly added, and the reaction mixture was stirred at -78 °C for 1 h. Subsequently, tributyl(chloro)stannane (5.22 g, 16.03 mmol, 4.31 mL) was slowly added to the reaction mixture, and the mixture was stirred at -78 °C for 1 h under nitrogen. The reaction mixture was quenched by slow addition of aqueous NH4Cl solution (20 mL) at 0 °C, and extracted with EtOAc (40 mLХ3). The combined organic layers were washed with brine (20 mL Х 2), dried over Na2SO4 and concentrated under reduced pressure to give the crude product tributyl-[3-(trideuteriomethyl)triazol-4-yl]stannane (T-1.4 g, 10.66 mmol) as a yellow oil. The obtained product was used in the next reaction without further purification.1 H NMR (400 MHz, DMSO-d6) δ ppm 7.60 (s, 1H), 1.49 (m, 4H), 1.35 - 1.21 (m, 8H), 1.19 - 1.09 (m, 6H), 0.88 - 0.83 (m, 9H).

[0312] Manufacturing Example 4: Tributyl-(3-ethyltriazol-4-yl)stanane (T-2)

[0313]

[0314] Tributyl-(3-ethyltriazol-4-yl)stanane (T-2, 2.4 g, 90.54% yield, 75% purity) was obtained as a pale yellow oil using the same method as in step 2 of Preparation Example 3 using commercially available 1-ethyl-1H-1,2,3-triazole. 1 H NMR (400 MHz, DMSO-d6) δ ppm 7.58 (s, 1H), 4.33 (q, J = 7.2 Hz, 2H), 1.37-1.21 (m, 12H), 1.20-1.13 (m, 6H), 0.86 (d, J = 8.0 Hz, 12H).

[0315] Manufacturing Example 5: Tributyl-(3-cyclopropyltriazol-4-yl)stannane (T-3)

[0316]

[0317] Step 1: Synthesis of 1-cyclopropyltriazole

[0318] To a solution of 4-methylbenzenesulfonohydrazide (5 g, 26.85 mmol) in methanol (50 mL) was added 2,2-dimethoxyacetaldehyde (4.89 g, 28.19 mmol, 4.25 mL). The reaction mixture was stirred at 25 °C for 2 h. Then, acetic acid (1.61 g, 26.85 mmol, 1.54 mL) and cyclopropanamine (1.53 g, 26.85 mmol, 1.86 mL) were added to the reaction mixture, and the mixture was stirred at 75 °C for 14 h. The reaction mixture was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography (10% EtOAc in petroleum ether) to give 1-cyclopropyltriazole (6 g, 49.15% yield, 90% purity) as a yellow oil. 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.13 (s, 1H), 7.69 (d, J = 0.8 Hz, 1H), 4.02 - 3.92 (m, 1H), 1.15 - 1.08 (m, 4H).

[0319] Step 2: Synthesis of tributyl-(3-cyclopropyltriazol-4-yl)stannane

[0320] A solution of 1-cyclopropyltriazole (200 mg, 1.83 mmol) in THF (4 mL) was cooled to -78 °C, n-BuLi (2.5 M, 1.10 mL) was slowly added, and the reaction mixture was stirred at -78 °C for 2 h. Subsequently, tributyl(chloro)stannane (1.14 g, 3.50 mmol, 942.15 μL) was slowly added to the reaction mixture, and the mixture was stirred at -78 °C for 1 h under nitrogen conditions. The reaction mixture was quenched by slowly adding an aqueous ammonium salt solution (10 mL) at 0 °C, and extracted with EtOAc (15 mLХ3). The combined organic layers were washed with brine (10 mL Х 2), dried over Na2SO4 and concentrated under reduced pressure to give tributyl-(3-cyclopropyltriazol-4-yl)stannane (T-3, 1 g, 95.93% yield, 70% purity) as a white oil. 1 H NMR (400 MHz, DMSO-d6) δ ppm 7.59 (s, 1H), 3.71 (tt, J = 3.6, 7.2 Hz, 1H), 1.53 - 1.46 (m, 6H), 1.29 - 1.27 (m, 6H), 1.10 - 1.05 (m, 10H), 0.87 (s, 9H).

[0321] Manufacturing Example 6: Tributyl-(3-isopropyltriazol-4-yl)stanane (T-4)

[0322]

[0323] Tributyl-(3-isopropyltriazol-4-yl)stanane (T-4, 1 g, 97.21% yield, 70% purity) was obtained as a white oil by the same method as step 2 of Preparation Example 3 using commercially available 1-isopropyl-1H-1,2,3-triazole. 1H NMR (400 MHz, DMSO-d6) δ ppm 7.55 (s, 1H), 4.52 (td, J = 6.8, 13.2 Hz, 1H), 1.48 (s, 12H), 1.29 (br d, J = 7.6 Hz, 6H), 1.14 - 1.09 (m, 6H), 0.86 (d, J = 7.6 Hz, 9H).

[0324] Manufacturing Example 7: 1-(2,2-difluorocyclopropyl)triazole (T-5)

[0325]

[0326] (+ / -)-1-(2,2-difluorocyclopropyl)triazole (T-5, 1.63 g, 33.47% yield, 80% purity) was obtained as a white oil using the same method as in step 1 of Preparation Example 5 using commercially available (+ / -)-2,2-difluorocyclopropanamine hydrogen chloride. 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.33 (s, 1H), 7.81 (d, J= 1.2 Hz, 1H), 4.93 - 4.82 (m, 1H), 2.65 - 2.51 (m, 2H); MS (ESI) m / z = 146.0 [M+H] + .

[0327] Manufacturing Example 8: 1-(3,3-difluorocyclobutyl)triazole (T-6)

[0328]

[0329] 1-(3,3-difluorocyclobutyl)triazole (T-6, 250 mg, 11.04% yield, 88.3% purity) was obtained as a white solid using the same method as in step 1 of Preparation Example 5 using commercially available 3,3-difluorocyclobutylamine hydrogen chloride. 1H NMR (400 MHz, DMSO-d6) δ ppm 8.29 (d, J = 0.4 Hz, 1H), 7.79 (d, J = 0.4 Hz, 1H), 5.34 - 5.09 (m, 1H), 3.31 - 3.18 (m, 4H); MS (ESI) m / z = 160.1 [M+H] + .

[0330] Manufacturing Example 9: Tributyl-[3-(1-methylcyclopropyl)triazol-4-yl]stanane (T-7)

[0331]

[0332] Step 1: Synthesis of 1-(1-methylcyclopropyl)-1H-1,2,3-triazole

[0333] 1-(1-methylcyclopropyl)-1H-1,2,3-triazole (50 mg, 0.7% yield) was obtained as a yellow oil using the same method as step 1 of Preparation Example 5 using commercially available 1-methylcyclopropylamine hydrogen chloride. 1 H NMR (400 MHz, CD3OD) δ ppm 8.07 (s, 1H), 7.69 (s, 1H), 1.70 (s, 3H), 1.38 - 1.34 (m, 2H), 1.12 - 1.07 (m, 2H); MS (ESI) m / z = 124.1 [M+H] + .

[0334] Step 2: Synthesis of tributyl-[3-(1-methylcyclopropyl)triazol-4-yl]stannane

[0335] Tributyl-[3-(1-methylcyclopropyl)triazol-4-yl]stanane (T-7, 167 mg, 99.79% yield) was obtained as a yellow oil using the same method as in step 2 of Preparation Example 5 using 1-(1-methylcyclopropyl)-1H-1,2,3-triazole. 1H NMR (400 MHz, DMSO-d6) δ ppm 7.58 (s, 1H), 1.55 (m, 6H), 1.52 (s, 3H), 1.30 - 1.29 (m, 6H), 1.17 (m, 6H), 1.03 (m, 4H), 0.87 - 0.87 (m, 9H).

[0336] Manufacturing Example 9-1: Tributyl-[3-(oxetan-3-yl)triazol-4-yl]stanane (T-8)

[0337]

[0338] Step 1: Synthesis of 1-(oxetan-3-yl)triazole

[0339] 1-(Oxetan-3-yl)triazole (1.4 g, 37.51% yield, 90% purity) was obtained as a yellow oil using the same method as in step 1 of Preparation Example 5 using commercially available oxetan-3-amine. 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.38 (d,J= 0.8 Hz, 1H), 7.82 (d,J= 0.8 Hz, 1H), 5.92 - 5.81 (m, 1H), 5.01 (t,J= 7.2 Hz, 2H), 4.92 - 4.88 (t,J= 7.2 Hz, 2H).

[0340] Step 2: Synthesis of tributyl-[3-(oxetan-3-yl)triazol-4-yl]stanane

[0341] Tributyl-[3-(oxetan-3-yl)triazol-4-yl]stanane (T-8, 600 mg, 90.64% yield) was obtained as a yellow oil using the same method as in step 2 of Preparation Example 5 using 1-(oxetan-3-yl)triazole. 1H NMR (400 MHz, DMSO-d6) δ ppm 7.67 (s, 1H), 5.64 - 5.53 (m, 1H), 5.00 (d, J = 6.8 Hz, 4H), 1.48 - 1.42 (m, 6H), 1.28 (br d, J = 7.6 Hz, 6H), 1.17 - 1.11 (m, 6H), 0.87 - 0.84 (m, 9H).

[0342] Manufacturing Example 9-2: Tributyl-[3-[(3R)-tetrahydrofuran-3-yl]triazol-4-yl]stanane (T-9)

[0343]

[0344] Step 1: Synthesis of 1-[(3R)-tetrahydrofuran-3-yl]triazole

[0345] 1-[(3R)-tetrahydrofuran-3-yl]triazole (140 mg, 2.97% yield, 79.34% purity) was obtained as a yellow oil using the same method as in step 1 of Preparation Example 5 using commercially available (3R)-tetrahydrofuran-3-amine hydrogen chloride. MS (ESI) m / z = 140.2 [M+H] + .

[0346] Step 2: Synthesis of tributyl-[3-[(3R)-tetrahydrofuran-3-yl]triazol-4-yl]stannane

[0347] Tributyl-[3-[(3R)-tetrahydrofuran-3-yl]triazol-4-yl]stanane (T-9,430 mg, 99.81% yield) was obtained as a yellow oil using the same method as step 2 of Manufacturing Example 5. 1 H NMR (400 MHz, DMSO-d6) δ ppm 7.61 (s, 1H), 5.08 - 5.00 (m, 1H), 4.13 - 4.05 (m, 2H), 3.94 - 3.80 (m, 2H), 2.46 - 2.40 (m, 1H), 2.34 - 2.23 (m, 1H), 1.59 (m, 6H), 1.30 (s, 6H), 1.20 - 1.16 (m, 6H), 0.85 (s, 9H).

[0348] Manufacturing Example 9-3: Tributyl-[3-[(3S)-tetrahydrofuran-3-yl]triazol-4-yl]stanane (T-10)

[0349]

[0350] Step 1: Synthesis of (1S)-1-tetrahydrofuran-3-yltriazole

[0351] (1S)-1-Tetrahydrofuran-3-yltriazole (280 mg, 6.94% yield, 92.61% purity) was obtained as a white oil in the same manner as in step 1 of Preparation Example 5 using commercially available (3S)-tetrahydrofuran-3-amine hydrochloride. 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.14 (d,J= 0.8 Hz, 1H), 7.74 (d,J= 0.8 Hz, 1H), 5.38-5.30 (m, 1H), 4.04-3.97 (m, 2H), 3.96-3.90 (m, 1H), 3.84 (dt,J= 5.6, 8.4 Hz, 1H), 2.50-2.44 (m, 1H), 2.34-2.24 (m, 1H); MS (ESI) m / z = 140.2 [M+H] + .

[0352] Step 2: Synthesis of tributyl-[3-[(3S)-tetrahydrofuran-3-yl]triazol-4-yl]stannane

[0353] Tributyl-[3-[(3S)-tetrahydrofuran-3-yl]triazol-4-yl] (T-10,460 mg, 99.66% yield) was obtained as a yellow oil using the same method as step 2 of Manufacturing Example 5. 1H NMR (400 MHz, DMSO-d6) δ ppm 7.61 (s, 1H), 5.08-5.00 (m, 1H), 4.12-4.06 (m, 2H), 3.94-3.84 (m, 2H), 2.47-2.41 (m, 1H), 2.29 (br dd,J= 3.2, 8.0 Hz, 1H), 1.58 (br d,J= 7.6 Hz, 6H), 1.47 (br d,J= 7.6 Hz, 6H), 1.19 (br d,J= 8.4 Hz, 6H), 1.13-1.07 (m, 9H).

[0354] Manufacturing Example 9-4: 1-[(1R, 2S)-2-fluorocyclopropyl]triazole (T-11)

[0355]

[0356] 1-[(1R, 2S)-2-fluorocyclopropyl] triazole (T-11,400 mg, 13.61% yield, 92.9% purity) was obtained as a yellow oil using the same method as in step 1 of Preparation Example 5 using commercially available (1R, 2S)-2-fluorocyclopropylamine hydrogen chloride. 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.21 (s, 1H), 7.75 (d,J= 1.2 Hz, 1H), 5.13 - 4.90 (m, 1H), 4.05 - 3.97 (m, 1H), 2.03 - 1.90 (m, 1H), 1.59 (dtd,J=6.0, 9.2, 16.4 Hz, 1H); MS (ESI) m / z = 128.0 [M+H] + .

[0357] Manufacturing Example 9-5: 1-[(1S, 2R)-2-fluorocyclopropyl]triazole (T-12)

[0358]

[0359] 1-[(1S, 2R)-2-fluorocyclopropyl]triazole (T-12,620 mg, 46.50% yield, 75% purity) was obtained as an oil using the same method as in step 1 of Preparation Example 5 using commercially available (1S, 2R)-2-fluorocyclopropylamine hydrogen chloride. 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.21 (s, 1H), 7.75 (d,J= 0.8 Hz, 1H), 5.14 - 4.90 (m, 1H), 4.06 - 3.96 (m, 1H), 2.03 - 1.90 (m, 1H), 1.66 - 1.51 (m, 1H); MS (ESI) m / z = 128.0 [M+H] + .

[0360] Manufacturing Example 9-6: 1-[(1S, 2S)-2-fluorocyclopropyl]triazole (T-13)

[0361]

[0362] 1-[(1S, 2S)-2-fluorocyclopropyl] triazole (T-13, 40 mg, 1.07% yield, 73.21% purity) was obtained as a yellow oil using the same method as in step 1 of Preparation Example 5 using commercially available (1S, 2S)-2-fluorocyclopropylamine hydrogen chloride. MS (ESI) m / z = 128.2 [M+H] + .

[0363] Manufacturing Example 9-7: 1-[(1R, 2R)-2-fluorocyclopropyl]triazole (T-14)

[0364]

[0365] 1-[(1R, 2R)-2-fluorocyclopropyl]triazole (T-14,800 mg, 26.37% yield, 90% purity) was obtained as a yellow oil using the same method as in step 1 of Preparation Example 5 using commercially available (1R, 2R)-2-fluorocyclopropylamine hydrogen chloride. 1H NMR (400 MHz, DMSO-d6) δ ppm 8.21 (s, 1H), 7.74 (d,J= 0.8 Hz, 1H), 5.37 - 5.15 (m, 1H), 4.59 - 4.46 (m, 1H), 1.92 - 1.79 (m, 1H), 1.78 - 1.68 (m, 1H); MS (ESI) m / z = 128.1 [M+H] + .

[0366] Manufacturing Example 9-8: 1-(thietan-3-yl)triazole (T-15)

[0367]

[0368] 1-(Thietan-3-yl)triazole (T-15, 3.67 g, 34.94% yield, 53.5% purity) was obtained as a yellow solid using the same method as in step 1 of Preparation Example 5 using commercially available thietan-3-amine hydrochloride. 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.33 (s, 1H), 7.77 (s, 1H), 5.98 (quin,J= 8.8 Hz, 1H), 3.95-3.87 (m, 2H), 3.52-3.45 (m, 2H); MS (ESI) m / z = 462.2 [M+H] + .

[0369] Manufacturing Example 10: [5-(3-methyltriazol-4-yl)-3-pyridyl]boronic acid (P-1)

[0370]

[0371] Step 1: Synthesis of 3-bromo-5-(3-methyltriazol-4-yl)pyridine

[0372] A solution of 3,5-dibromopyridine (10.92 g, 46.10 mmol), commercially available tributyl-(3-methyltriazol-4-yl)stannane (22.3 g, 59.92 mmol) and Pd(PPh3)2Cl2 (3.24 g, 4.61 mmol) in toluene (200 mL) was purged with nitrogen three times and stirred at 90 °C under nitrogen for 16 h. Water (150 mL) was poured into the reaction mixture and extracted with EtOAc (200 mL Х 3). The combined organic layers were washed with brine (200 mL Х 3), dried over Na2SO4 and concentrated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (80% EA in petroleum ether) to give 3-bromo-5-(3-methyltriazol-4-yl)pyridine (4.4 g, 39.93% yield) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ ppm8.83-8.83 (m, 1H), 8.40 (t, J= 2.0 Hz, 1H), 8.09 (s, 1H), 7.66-7.49 (m, 1H), 4.13 (s, 3H); MS (ESI) m / z = 153.0 [M+H] + .

[0373] Step 2: Synthesis of [5-(3-methyltriazol-4-yl)-3-pyridyl]boronic acid

[0374] A solution of 3-bromo-5-(3-methyltriazol-4-yl)pyridine (6.76 g, 28.28 mmol), B2pin2 (14.36 g, 56.55 mmol), Pd(dppf)Cl2 (2.31 g, 2.83 mmol), and KOAc (5.55 g, 56.55 mmol) in dioxane (120 mL) was purged with nitrogen three times and stirred at 100 °C under nitrogen for 16 h. The reaction mixture was filtered and concentrated under reduced pressure to give the crude product, which was purified by silica gel column chromatography (3% MeOH in DCM) to give [5-(3-methyltriazol-4-yl)-3-pyridyl]boronic acid (P-1, 6.76 g, 78.52% yield, 67% purity) as a dark brown solid. MS (ESI) m / z = 205.1 [M+H] + .

[0375] Manufacturing Example 11: 1-(5-Bromopyridin-3-yl)-6-oxo-1,6-dihydropyridazine-3-carboxylic acid (3-1)

[0376]

[0377] Step 1: Synthesis of methyl 1-(5-bromopyridin-3-yl)-6-oxo-1,6-dihydropyridazine-3-carboxylate

[0378] A solution of methyl 6-oxo-1H-pyridazine-3-carboxylate (C-1, 5 g, 32.44 mmol), 3-bromo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (9.21 g, 32.44 mmol), Cu(OAc)2 (2.95 g, 16.22 mmol), boric acid (4.01 g, 64.88 mmol) and 4A MS (1 g) in MeCN (120 mL) was replaced with oxygen three times and stirred at 90 °C under oxygen conditions for 16 h. Water (30 mL) was poured into the reaction mixture, and the mixture was extracted with EtOAc (30 mL Х 3). The combined organic layer was washed with brine (50 mL), dried over Na2SO4, and concentrated under reduced pressure to give the crude product, which was purified by silica gel column chromatography (58% DCM in petroleum ether) to give methyl 1-(5-bromopyridin-3-yl)-6-oxo-1,6-dihydropyridazine-3-carboxylate (2-1, 2.2 g, 21.54% yield, 98.49% purity) as a white solid. MS (ESI) m / z = 312.0 [M+H] + .

[0379] Step 2: Synthesis of 1-(5-bromopyridin-3-yl)-6-oxo-1,6-dihydropyridazine-3-carboxylic acid

[0380] A solution of methyl 1-(5-bromopyridin-3-yl)-6-oxo-1,6-dihydropyridazine-3-carboxylate (2-1, 30 g, 96.74 mmol) and NaOH (5.80 g, 145.11 mmol) in THF (600 mL) and H2O (80 mL) was degassed and purged with nitrogen three times, and stirred under nitrogen at 20 °C for 30 min. The reaction mixture was adjusted to pH = 6-7 with 1 N HCl solution and concentrated under reduced pressure to give 1-(5-bromopyridin-3-yl)-6-oxo-1,6-dihydropyridazine-3-carboxylic acid (3-1, 28 g, 90.91% yield, 93% purity) as a yellow solid. MS (ESI) m / z = 295.7 [M+H] + .

[0381] Manufacturing Example 12: 1-(5-Bromopyridin-3-yl)-5-methyl-6-oxo-1,6-dihydropyridazine-3-carboxylic acid (3-2)

[0382]

[0383] Step 1: Synthesis of methyl 1-(5-bromopyridin-3-yl)-5-methyl-6-oxo-1,6-dihydropyridazine-3-carboxylate

[0384] A solution of methyl 5-methyl-6-oxo-1H-pyridazine-3-carboxylate (C-2, 600 mg, 3.57 mmol), 3-bromo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (1.01 g, 3.57 mmol), Cu(OAc)2 (1.30 g, 7.14 mmol), and pyridine (564.50 mg, 7.14 mmol, 576.02 μL) in MeCN (15 mL) was replaced with oxygen three times and stirred at 90 °C under oxygen conditions for 16 h. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (80% EtOAc in petroleum ether) to give methyl 1-(5-bromopyridin-3-yl)-5-methyl-6-oxo-1,6-dihydropyridazine-3-carboxylate (2-2,200 mg, 13.83% yield, 80% purity) as a yellow solid. MS (ESI) m / z = 325.9 [M+H] + .

[0385] Step 2: Synthesis of 1-(5-bromopyridin-3-yl)-5-methyl-6-oxo-1,6-dihydropyridazine-3-carboxylic acid

[0386] Using methyl 1-(5-bromopyridin-3-yl)-5-methyl-6-oxo-1,6-dihydropyridazine-3-carboxylate (2-2), the same method as in step 2 of Preparation Example 11 was used to obtain 1-(5-bromopyridin-3-yl)-5-methyl-6-oxo-1,6-dihydropyridazine-3-carboxylic acid (3-2, 140 mg, 83.74% yield, 95% purity) as a white solid. MS (ESI) m / z = 311.7 [M+H]+.

[0387] Manufacturing Example 13: 6-Oxo-1-[5-[3-(trideuteriomethyl)triazol-4-yl]-3-pyridyl]pyridazine-3-carboxylic acid (5-1)

[0388]

[0389] Step 1: Synthesis of methyl 6-oxo-1-[5-[3-(trideuteriomethyl)triazol-4-yl]-3-pyridyl]pyridazine-3-carboxylate

[0390] A solution of methyl 1-(5-bromopyridin-3-yl)-6-oxo-1,6-dihydropyridazine-3-carboxylate (2-1, 1 g, 3.22 mmol), tributyl-[3-(trideuteriomethyl)triazol-4-yl]stannane (T-1, 3.63 g, 9.67 mmol) and Pd(PPh3)2Cl2 (226.34 mg, 322.47 μmol) in toluene (10 mL) was purged with nitrogen three times and stirred under nitrogen at 100 °C for 16 h. Water (20 mL) was poured into the reaction mixture and extracted with EtOAc (30 mL Х 3). The combined organic layers were washed with brine (20 mL Х 2), dried over Na2SO4 and concentrated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography (5% MeOH in DCM) to give methyl 6-oxo-1-[5-[3-(trideuteriomethyl)triazol-4-yl]-3-pyridyl]pyridazine-3-carboxylate (420 mg, 30.57% yield, 74% purity) as a yellow solid. MS (ESI) m / z = 316.1 [M + H] + .

[0391] Step 2: Synthesis of 6-oxo-1-[5-[3-(trideuteriomethyl)triazol-4-yl]-3-pyridyl]pyridazine-3-carboxylic acid

[0392] To a solution of methyl 6-oxo-1-[5-[3-(trideuteriomethyl)triazol-4-yl]-3-pyridyl]pyridazine-3-carboxylate (420 mg, 1.33 mmol) in THF (8 mL) and H2O (2 mL) was added NaOH (79.92 mg, 2.00 mmol). The solution was purged with nitrogen three times, and the mixture was stirred under nitrogen at 25 °C for 2 h. The mixture was adjusted to pH = 6-7 with 1 N HCl aqueous solution, and then concentrated under reduced pressure to give 6-oxo-1-[5-[3-(trideuteriomethyl)triazol-4-yl]-3-pyridyl]pyridazine-3-carboxylic acid (5-1, 420 mg, 65.93% yield, 63% purity) as a yellow solid. MS (ESI) m / z = 302.1 [M + H] + .

[0393] Manufacturing Example 14: Methyl 5-bromo-1-(5-(1-methyl-1H-1,2,3-triazol-5-yl)pyridin-3-yl)-6-oxo-1,6-dihydropyridazine-3-carboxylate (4-1)

[0394]

[0395] To a solution of [5-(3-methyltriazol-4-yl)-3-pyridyl]boronic acid (P-1,200 mg, 872.58 μmol) in MeCN (10 mL) were added pyridine (414.13 mg, 5.24 mmol, 422.58 μL), TEA (530.29 mg, 5.24 mmol, 730.43 μL), Cu(OAc)2 (158.49 mg, 872.58 μmol), and methyl 5-bromo-6-oxo-1H-pyridazine-3-carboxylate (C-3, 251.54 mg, 1.05 mmol). The reaction mixture was stirred at 90 °C for 16 h under oxygen conditions. The mixture was filtered and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then purified by silica gel column chromatography (2% MeOH in DCM) to obtain methyl 5-bromo-1-(5-(1-methyl-1H-1,2,3-triazol-5-yl)pyridin-3-yl)-6-oxo-1,6-dihydropyridazine-3-carboxylate (4-1, 170 mg, 36.86% yield, 74% purity) as a yellow solid. MS (ESI) m / z = 392.9 [M + H] + .

[0396] Manufacturing Example 15: 1-[5-(3-methyltriazol-4-yl)-3-pyridyl]-6-oxo-5-(trifluoromethyl)pyridazine-3-carboxylic acid (5-2)

[0397]

[0398] Step 1: Synthesis of methyl 1-[5-(3-methyltriazol-4-yl)-3-pyridyl]-6-oxo-5-(trifluoromethyl)pyridazine-3-carboxylate

[0399] To a solution of methyl 5-bromo-1-(5-(1-methyl-1H-1,2,3-triazol-5-yl)pyridin-3-yl)-6-oxo-1,6-dihydropyridazine-3-carboxylate (4-1, 300 mg, 766.91 μmol) in DMF (5 mL) were added CuI (14.61 mg, 76.69 μmol) and methyl 2,2-difluoro-2-fluorosulfonyl-acetate (736.67 mg, 3.83 mmol, 487.86 μL). The mixture was stirred at 80 °C for 3 h. The reaction mixture was concentrated under reduced pressure to give the crude product. This was diluted with EtOAc (30 mL) and washed with brine (15 mL Х 3). The organic layer was dehydrated with anhydrous Na2SO4 and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (1% MeOH in DCM) to obtain methyl 1-[5-(3-methyltriazol-4-yl)-3-pyridyl]-6-oxo-5-(trifluoromethyl)pyridazine-3-carboxylate (4-2, 40 mg, 9.59% yield, 69.9% purity) as a yellow oil. MS (ESI) m / z = 380.9 [M+H] + .

[0400] Step 2: Synthesis of 1-[5-(3-methyltriazol-4-yl)-3-pyridyl]-6-oxo-5-(trifluoromethyl)pyridazine-3-carboxylic acid

[0401] Using methyl 1-[5-(3-methyltriazol-4-yl)-3-pyridyl]-6-oxo-5-(trifluoromethyl)pyridazine-3-carboxylate (4-2), 1-[5-(3-methyltriazol-4-yl)-3-pyridyl]-6-oxo-5-(trifluoromethyl)pyridazine-3-carboxylic acid (5-2, 30 mg, 27.26% yield, 35% purity) was obtained as a yellow solid. MS (ESI) m / z = 366.9 [M+H] + .

[0402] Manufacturing Example 16: 1-[5-(3-Cyclopropyltriazol-4-yl)-3-pyridyl]-6-oxo-pyridazine-3-carboxylic acid (5-3)

[0403]

[0404] Using methyl 1-(5-bromopyridin-3-yl)-6-oxo-1,6-dihydropyridazine-3-carboxylate (2-1) and tributyl-(3-cyclopropyltriazol-4-yl)stannane (T-3), 1-[5-(3-cyclopropyltriazol-4-yl)-3-pyridyl]-6-oxo-pyridazine-3-carboxylic acid (5-3, 35 mg, 88.82% yield, 97.3% purity) was obtained as a yellow solid in the same manner as in Preparation Example 13. 1 H NMR (400 MHz, DMSO-d6) δ ppm 9.03 (d,J= 2.0 Hz, 1H), 8.95 (d,J= 2.4 Hz, 1H), 8.56 (t,J= 2.0 Hz, 1H), 8.17 (s, 1H), 7.96 (d,J= 9.6 Hz, 1H), 7.22 (d,J= 9.6 Hz, 1H), 4.01 (td,J= 3.6, 7.3 Hz, 2H), 1.20 - 1.15 (m, 2H), 1.12 (br d,J= 2.8 Hz, 2H); MS (ESI) m / z = 324.9 [M+H] + .

[0405] Manufacturing Example 16-1: 1-[5-(3-methyltriazol-4-yl)-3-pyridyl]-6-oxo-pyridazine-3-carboxylic acid (5-4)

[0406]

[0407] Step 1: Synthesis of methyl 1-[5-(3-methyltriazol-4-yl)-3-pyridyl]-6-oxo-pyridazine-3-carboxylate

[0408] A solution of methyl 6-oxo-1H-pyridazine-3-carboxylate (C-1, 3.78 g, 24.51 mmol), [5-(3-methyltriazol-4-yl)-3-pyridyl]boronic acid (P-1, 5 g, 24.51 mmol), Cu(OAc)2 (4.45 g, 24.51 mmol), TEA (4.96 g, 49.02 mmol, 6.82 mL), and pyridine (3.88 g, 49.02 mmol, 3.96 mL) in MeCN (100 mL) was replaced with oxygen three times and stirred at 90 °C under oxygen conditions for 16 h. The reaction mixture was filtered and concentrated under reduced pressure to obtain the crude product, which was purified by silica gel column chromatography (3% MeOH in DCM) to obtain methyl 1-[5-(3-methyltriazol-4-yl)-3-pyridyl]-6-oxo-pyridazine-3-carboxylate (1.83 g, 5.27 mmol, 21.51% yield, 89.98% purity) as a dark brown solid. MS (ESI) m / z = 312.9 [M+H] + .

[0409] Step 2: Synthesis of 1-[5-(3-methyltriazol-4-yl)-3-pyridyl]-6-oxo-pyridazine-3-carboxylic acid

[0410] To a solution of methyl 1-[5-(3-methyltriazol-4-yl)-3-pyridyl]-6-oxo-pyridazine-3-carboxylate (1.0 g, 3.20 mmol) in THF (4 mL) and H2O (2 mL) was added NaOH (256.16 mg, 6.40 mmol). After three nitrogen purges, the mixture was stirred at 25 °C under nitrogen for 16 h. The mixture was adjusted to pH = 2-3 with 1 N HCl aqueous solution, and the solid was filtered to obtain 1-[5-(3-methyltriazol-4-yl)-3-pyridyl]-6-oxo-pyridazine-3-carboxylic acid (5-4,725 mg, 74.01% yield, 97.5% purity) as a yellow solid. MS (ESI) m / z = 299.0 [M+H] + .

[0411] Manufacturing Example 17: (R)-5'-Bromo-N-(1-(2-fluoro-3-(1,1,2-trifluoroethyl)phenyl)ethyl)-2-oxo-2H-[1,3'-bipyridine]-5-carboxamide (D-1)

[0412]

[0413] Step 1: Synthesis of methyl 1-(5-bromopyridin-3-yl)-6-oxo-pyridine-3-carboxylate

[0414] A solution of 5-bromopyridin-3-amine (24.70 g, 142.75 mmol) and methyl 6-oxopyran-3-carboxylate (20 g, 129.77 mmol) in pyridine (250 mL) was degassed and filled with nitrogen three times, and the mixture was stirred at 80 °C for 16 h under nitrogen conditions. The mixture was concentrated under reduced pressure and purified by flash silica gel chromatography (ISCO ® ; 120 g SepaFlash ® The crude product (10 g) was purified by silica flash column, 100% DCM gradient elution at 90 mL / min. The crude product was triturated in DCM at 20 °C for 10 min to give methyl 1-(5-bromopyridin-3-yl)-6-oxo-pyridine-3-carboxylate (2-3, 3.2 g, 7.61% yield, 95.45% purity) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.83 (d, J = 2.0 Hz, 1H), 8.71 (d, J = 2.0 Hz, 1H), 8.47 (d, J = 2.4 Hz, 1H), 8.37 (t, J = 2.0 Hz, 1H), 7.92 (dd, J = 2.4, 9.6 Hz, 1H), 6.59 (d, J = 9.6 Hz, 1H), 3.80 (s, 3H); MS (ESI) m / z = 308.9 [M+H] + .

[0415] Step 2: Synthesis of 1-(5-bromopyridin-3-yl)-6-oxo-pyridine-3-carboxylic acid

[0416] 1-(5-bromopyridin-3-yl)-6-oxo-pyridine-3-carboxylic acid (3-3.3 g, crude product) was obtained as a yellow solid using the same method as in step 2 of Preparation Example 13 using methyl 1-(5-bromopyridin-3-yl)-6-oxo-pyridine-3-carboxylate (2-3). MS (ESI) m / z = 297.0 [M+H] + .

[0417] Step 3: Synthesis of (R)-5'-bromo-N-(1-(2-fluoro-3-(1,1,2-trifluoroethyl)phenyl)ethyl)-2-oxo-2H-[1,3'-bipyridine]-5-carboxamide

[0418] A solution of 1-(5-bromopyridin-3-yl)-6-oxo-pyridine-3-carboxylic acid (3-3, 600 mg, 2.03 mmol), (1R)-1-[2-fluoro-3-(1,1,2-trifluoroethyl)phenyl]ethanamine (A-2, 419.11 mg, 1.63 mmol, HCl), HOBt (549.49 mg, 4.07 mmol), EDCI (779.57 mg, 4.07 mmol), and DIEA (788.36 mg, 6.10 mmol, 1.06 mL) in DMF (8 mL) was stirred at 40 °C for 1 h. The reaction mixture was poured into water (30 mL) and extracted with EtOAc (30 mL Х 3). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure to give the crude product. The residue was purified by silica gel column chromatography (2% MeOH in DCM) to give (R)-5'-bromo-N-(1-(2-fluoro-3-(1,1,2-trifluoroethyl)phenyl)ethyl)-2-oxo-2H-[1,3'-bipyridine]-5-carboxamide (D-1, 800 mg, 70.40% yield, 89.16% purity) as a yellow solid. MS (ESI) m / z = 499.9 [M+H] +.

[0419] Manufacturing Example 18: 1-(5-Bromopyridin-3-yl)-N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethyl]-6-oxo-pyridine-3-carboxamide (D-2)

[0420]

[0421] 1-(5-Bromopyridin-3-yl)-6-oxo-pyridine-3-carboxylic acid (3-3) and (1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethanamine hydrogen chloride (A-1) were used in the same manner as in step 3 of Preparation Example 17 to obtain 1-(5-bromopyridin-3-yl)-N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethyl]-6-oxo-pyridine-3-carboxamide (D-2, 1.03 g, 63.68% yield, 96% purity) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ ppm 8.77 (d,J= 1.6 Hz, 1 H), 8.60 (d,J= 2.0 Hz, 1 H), 8.06 (d,J= 2.4 Hz, 1 H), 7.99 (t,J= 2.0 Hz, 1 H), 7.70 (dd,J= 9.6, 2.8 Hz, 1 H), 7.38 - 7.52 (m, 2 H), 7.15 - 7.22 (m, 1 H), 6.68 (d,J= 9.6 Hz, 1 H), 6.35 (br d,J=7.6 Hz, 1 H), 5.45 (quin,J= 7.2 Hz, 1 H), 2.06 (d,J= 0.8 Hz, 0.75 H), 2.01 (d,J= 0.8 Hz, 1.5 H), 1.97 (d,J= 0.8 Hz, 0.75 H), 1.63 (d,J= 7.2 Hz, 3 H); MS (ESI) m / z = 480.1 [M+H] + .

[0422] Manufacturing Example 19: 1-(5-Bromopyridin-3-yl)-N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethyl]-6-oxo-pyridazine-3-carboxamide (D-3)

[0423]

[0424] Using 1-(5-bromopyridin-3-yl)-6-oxo-1,6-dihydropyridazine-3-carboxylic acid (3-1) and (1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethanamine (A-1), 1-(5-bromopyridin-3-yl)-N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethyl]-6-oxo-pyridazine-3-carboxamide (D-3, 590 mg, 32.26% yield, 80% purity) was obtained as a yellow oil. MS (ESI) m / z = 481.0 [M+H] + .

[0425] Manufacturing Example 20: (R)-1-(5-bromopyridin-3-yl)-N-(1-(2-fluoro-3-(1,1,2-trifluoroethyl)phenyl)ethyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide (D-4)

[0426]

[0427] Using 1-(5-bromopyridin-3-yl)-6-oxo-1,6-dihydropyridazine-3-carboxylic acid (3-1) and (1R)-1-[2-fluoro-3-(1,1,2-trifluoroethyl)phenyl]ethanamine (A-2), (R)-1-(5-bromopyridin-3-yl)-N-(1-(2-fluoro-3-(1,1,2-trifluoroethyl)phenyl)ethyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide (D-4, 1.1 g, 70.15% purity, 86.03% yield) was obtained as a yellow oil in the same manner as in step 3 of Preparation Example 17. MS (ESI) m / z = 500.9 [M+H] + .

[0428] Manufacturing Example 21: (R)-1-(5-Bromopyridin-3-yl)-N-(1-(3-(1,1-difluoroethyl)-2-fluorophenyl)ethyl)-5-methyl-6-oxo-1,6-dihydropyridazine-3-carboxamide (D-5)

[0429]

[0430] Using 1-(5-bromopyridin-3-yl)-5-methyl-6-oxo-1,6-dihydropyridazine-3-carboxylic acid (3-2) and (1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethanamine (A-1), (R)-1-(5-bromopyridin-3-yl)-N-(1-(3-(1,1-difluoroethyl)-2-fluorophenyl)ethyl)-5-methyl-6-oxo-1,6-dihydropyridazine-3-carboxamide (D-5, 260 mg, 81.05% yield, 94.6% purity) was obtained as a yellow oil in the same manner as in step 3 of Preparation Example 17. MS (ESI) m / z = 476.8 [M+H] + .

[0431] Manufacturing Example 22: (R)-1-(5-Bromopyridin-3-yl)-N-(1-(2-fluoro-3-(1,1,2-trifluoroethyl)phenyl)ethyl)-5-methyl-6-oxo-1,6-dihydropyridazine-3-carboxamide (D-6)

[0432]

[0433] Using 1-(5-bromopyridin-3-yl)-5-methyl-6-oxo-1,6-dihydropyridazine-3-carboxylic acid (3-2) and (1R)-1-[2-fluoro-3-(1,1,2-trifluoroethyl)phenyl]ethanamine (A-2), (R)-1-(5-bromopyridin-3-yl)-N-(1-(2-fluoro-3-(1,1,2-trifluoroethyl)phenyl)ethyl)-5-methyl-6-oxo-1,6-dihydropyridazine-3-carboxamide (D-6, 117 mg, 33.14% yield, 93.78% purity) was obtained as a yellow oil in the same manner as in step 3 of Preparation Example 17. MS (ESI) m / z = 513.1 [M+H] + .

[0434] Example 1: (R)-1-(5-(1-Cyclopropyl-1H-1,2,3-triazol-5-yl)pyridin-3-yl)-N-(1-(3-(1,1-difluoroethyl)-2-fluorophenyl)ethyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide

[0435]

[0436] A solution of 1-(5-bromopyridin-3-yl)-N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethyl]-6-oxo-pyridazine-3-carboxamide (D-3, 60 mg, 124.67 μmol), tributyl-(3-cyclopropyltriazol-4-yl)stannane (T-3, 99.28 mg, 249.34 μmol) and Pd(PPh3)2Cl2 (8.75 mg, 12.47 μmol) in dioxane (3 mL) was filled with nitrogen three times, and the mixture was stirred at 100 °C for 12 h under nitrogen conditions. The mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The first purification was performed by column chromatography (2% MeOH in DCM), and the residue dissolved in MeOH (5 mL) was purified by preparative HPLC (column: FPrepulite XP tC 18 40*200mm*7um; mobile phase: [water (NH3H2O ​​+ NH4HCO3)-MeCN]; gradient: 22%-62% B for 20 min), and MeCN was removed under reduced pressure. The residual solvent was removed by lyophilization to obtain the compound of Example 1 (21.3 mg, 33.52% yield, 99.95% purity) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ ppm 9.12 (d, J = 2.4 Hz, 1H), 9.07 (d, J = 8.0 Hz, 1H), 9.02 (d, J = 2.0 Hz, 1H), 8.65 (t, J = 2.0 Hz, 1H), 8.17 (s, 1H), 7.92 (d, J = 9.6 Hz, 1H), 7.64 (t, J = 7.2 Hz, 1H), 7.49 - 7.42 (m, 1H), 7.28 (t, J = 7.6 Hz, 1H), 7.22 (d, J = 9.6 Hz, 1H), 5.42 (quin, J = 7.2 Hz, 1H), 4.08 - 3.98 (m, 1H), 2.01 (t, J = 19.2 Hz, 3H), 1.49 (d, J = 7.2 Hz, 3H), 1.19 - 1.12 (m, 4H); MS (ESI) m / z = 510.6 [M+H] + .

[0437] Examples 2 to 31

[0438] The compounds in Table 1 below were prepared through a one-step Stille coupling using intermediates corresponding to the structure of the target compound based on the manufacturing example in the same manner as Example 1.

[0439]

[0440]

[0441]

[0442]

[0443]

[0444]

[0445]

[0446]

[0447] Example 27: (R)-1-(5-(1-(1,1-deoxidothietan-3-yl)-1H-1,2,3-triazol-5-yl)pyridin-3-yl)-N-(1-(2-fluoro-3-(trifluoromethyl)phenyl)ethyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide

[0448]

[0449] Step 1: 1-(5-Bromo-3-pyridyl)-N-[(1R)-1-[2-fluoro-3-(trifluoromethyl)phenyl]ethyl]-6-oxo-pyridazine-3-carboxamide

[0450] Using 1-(5-bromopyridin-3-yl)-6-oxo-1,6-dihydropyridazine-3-carboxylic acid (3-1) and (1R)-1-[2-fluoro-3-(trifluoromethyl)phenyl]ethanamine hydrogen chloride (A-4), 1-(5-bromo-3-pyridyl)-N-[(1R)-1-[2-fluoro-3-(trifluoromethyl)phenyl]ethyl]-6-oxo-pyridazine-3-carboxamide was obtained as a yellow solid in the same manner as in step 3 of Preparation Example 17.

[0451] Step 2: N-[(1R)-1-[2-fluoro-3-(trifluoromethyl)phenyl]ethyl]-6-oxo-1-[5-[3-(thietan-3-yl)triazol-4-yl]-3-pyridyl]pyridazine-3-carboxamide

[0452] A mixture of 1-(5-bromo-3-pyridyl)-N-[(1R)-1-[2-fluoro-3-(trifluoromethyl)phenyl]ethyl]-6-oxo-pyridazine-3-carboxamide (100 mg, 206.09 μmol), 1-(thietan-3-yl)triazole (T-15, 145.49 mg, 1.03 mmol), PPh3 (5.41 mg, 20.61 μmol), Pd(OAc)2 (9.25 mg, 41.22 μmol), Cs2CO3 (201.44 mg, 618.26 μmol), and Cu(OTf)2 (7.45 mg, 20.61 μmol) in toluene (1.5 mL) was degassed and purged with N2 three times, and the mixture was stirred at 110 °C for 12 h. The mixture was stirred under a nitrogen atmosphere. The reaction mixture was added to water (30 mL) and extracted with EtOAc (30 mL Х 3). The combined organic layers were washed with brine (50 mL), dehydrated over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by prep-HPLC (column: F-Prepulite XP tC18 40*200mm*7um; mobile phase: [H2O (0.05% NH3H2O+10mM NH4HCO3)-ACN]; gradient: 26%-66% B, 20.0 min) to obtain N-[(1R)-1-[2-fluoro-3-(trifluoromethyl)phenyl]ethyl]-6-oxo-1-[5-[3-(thietan-3-yl)triazol-4-yl]-3-pyridyl]pyridazine-3-carboxamide (4.7 mg, 4.16% yield, 94.45% purity) as a white solid. 1H NMR (400 MHz, CDCl3) δ ppm 9.03 (d,J=2.4 Hz, 1 H), 8.69 (d,J=2.0 Hz, 1 H), 8.17 (t,J= 2.4 Hz, 1 H), 8.09 (d,J= 9.6 Hz, 1 H), 7.86 (s, 1 H), 7.57 (t,J= 7.2 Hz, 2 H), 7.42 (br d,J= 8.4 Hz, 1 H), 7.22 - 7.26 (m, 1 H), 7.17 (d,J= 9.6 Hz, 1 H), 5.66 - 5.77 (m, 1 H), 5.45 (quin,J= 7.2 Hz, 1 H), 4.34 (td,J= 9.2, 1.6 Hz, 2 H), 3.40 - 3.45 (m, 2 H), 1.66 (d,J= 7.2 Hz, 3 H); MS (ESI) m / z = 546.2 [M+H] + .

[0453] Step 3: (R)-1-(5-(1-(1,1-deoxidothietan-3-yl)-1H-1,2,3-triazol-5-yl)pyridin-3-yl)-N-(1-(2-fluoro-3-(trifluoromethyl)phenyl)ethyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide

[0454] A mixture of N-[(1R)-1-[2-fluoro-3-(trifluoromethyl)phenyl]ethyl]-6-oxo-1-[5-[3-(thietan-3-yl)triazol-4-yl]-3-pyridyl]pyridazine-3-carboxamide (5 mg, 9.17 μmol) and oxone (3.08 mg, 18.33 μmol) in THF (0.5 mL) and H2O (0.15 mL) was degassed and purged three times with N2, and the mixture was stirred at 25 °C for 12 h under a nitrogen atmosphere. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by prep-HPLC (column: F-Prepulite XP tC18 40*200mm*7um; mobile phase: [H2O (0.05% NH3H2O ​​+ 10 mM NH4HCO3) - ACN]; 16%-56% B, 20.0 min) to obtain the compound of Example 27 (2.4 mg, 44.89% yield, 99.0% purity) as a white solid. 1 H NMR (400 MHz, CDCl3)δppm 9.06 (d,J= 2.0 Hz, 1H), 8.71 (s, 1H), 8.21 (s, 1H), 8.10 (d,J= 9.6 Hz, 1H), 7.94 (s, 1H), 7.61 - 7.53 (m, 2H), 7.45 (br d,J= 8.0 Hz, 1H), 7.26 - 7.23 (m, 1H), 7.17 (d,J= 9.6 Hz, 1H), 5.45 (quin,J= 7.2 Hz, 1H), 5.37 - 5.28 (m, 1H), 5.11 - 5.04 (m, 2H), 4.77 - 4.69 (m, 2H), 1.67 (br d,J= 7.2 Hz, 3H); MS (ESI) m / z = 578.1 [M+H] + .

[0455] Example 28: (R)-N-(1-(3-(1,1-difluoroethyl)-2-fluorophenyl)ethyl)-1-(5-(1-(1,1-deoxidothietan-3-yl)-1H-1,2,3-triazol-5-yl)pyridin-3-yl)-6-oxo-1,6-dihydropyridazine-3-carboxamide

[0456]

[0457] Step 1: N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluoro-phenyl]ethyl]-6-oxo-1-[5-[3-(thietan-3-yl)triazol-4-yl]-3-pyridyl]pyridazine-3-carboxamide

[0458] Using 1-(5-bromopyridin-3-yl)-N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethyl]-6-oxo-pyridazine-3-carboxamide (D-3) and 1-(thietan-3-yl)triazole (T-15), N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluoro-phenyl]ethyl]-6-oxo-1-[5-[3-(thietan-3-yl)triazol-4-yl]-3-pyridyl]pyridazine-3-carboxamide (6.1 mg, 12.87% yield, 92.2% purity) was obtained as a white solid in the same manner as in step 2 of Preparation Example 27. 1 H NMR (400 MHz, CDCl3)δppm 9.04 (br d,J= 1.2 Hz, 1H), 8.68 (br s, 1H), 8.17 (s, 1H), 8.09 (d,J= 9.6 Hz, 1H), 7.86 (s, 1H), 7.51 - 7.46 (m, 1H), 7.45 - 7.39 (m, 2H), 7.22 - 7.15 (m, 2H), 5.71 (quin,J= 8.8 Hz, 1H), 5.46 (quin,J= 7.2 Hz, 1H), 4.34 (t,J= 9.2 Hz, 2H), 3.46 - 3.36 (m, 2H), 2.05 - 1.94 (m, 3H), 1.64 (d,J= 7.2 Hz, 3H); MS (ESI) m / z = 542.6 [M+H] + .

[0459] Step 2: (R)-N-(1-(3-(1,1-difluoroethyl)-2-fluorophenyl)ethyl)-1-(5-(1-(1,1-deoxidothietan-3-yl)-1H-1,2,3-triazol-5-yl)pyridin-3-yl)-6-oxo-1,6-dihydropyridazine-3-carboxamide

[0460] Using N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluoro-phenyl]ethyl]-6-oxo-1-[5-[3-(thietan-3-yl)triazol-4-yl]-3-pyridyl]pyridazine-3-carboxamide, the compound of Example 28 (3.0 mg, 62.75% yield, 99.68% purity) was obtained as a white solid in the same manner as in Step 3 of Preparation Example 27. 1 H NMR (400 MHz, CDCl3)δppm 9.06 (d,J= 2.0 Hz, 1H), 8.70 (d,J= 2.0 Hz, 1H), 8.20 (t,J= 2.0 Hz, 1H), 8.11 (d,J= 9.6 Hz, 1H), 7.93 (s, 1H), 7.52 - 7.40 (m, 3H), 7.22 - 7.15 (m, 2H), 5.50 - 5.42 (m, 1H), 5.37 - 5.27 (m, 1H), 5.11 - 5.03 (m, 2H), 4.77 - 4.69 (m, 2H), 2.00 (t,J= 18.8 Hz, 3H), 1.65 (d,J= 7.2 Hz, 3H); MS (ESI) m / z = 574.1 [M+H] + .

[0461] Example 29: (R)-1-(5-(1-(1,1-deoxidothietan-3-yl)-1H-1,2,3-triazol-5-yl)pyridin-3-yl)-N-(1-(2-fluoro-3-(1,1,2-trifluoroethyl)phenyl)ethyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide

[0462]

[0463] (R)-1-(5-(1-(1,1-deoxidothietan-3-yl)-1H-1,2,3-triazol-5-yl)pyridin-3-yl)-N-(1-(2-fluoro-3-(1,1,2-trifluoroethyl)phenyl)ethyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide is (R)-1-(5-bromopyridin-3-yl)-N-(1-(2-fluoro-3-(1,1,2-trifluoroethyl)phenyl)ethyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide (D-4) and

[0464] 1-(thietan-3-yl)triazole (T-15) can be synthesized using the same method as steps 2 and 3 of Example 27.

[0465] Example 30: (R)-1-(5-(1-(1,1-deoxidothietan-3-yl)-1H-1,2,3-triazol-5-yl)pyridin-3-yl)-N-(1-(2-methyl-3-(trifluoromethyl)phenyl)ethyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide

[0466]

[0467] (R)-1-(5-(1-(1,1-Dioxidothietan-3-yl)-1H-1,2,3-triazol-5-yl)pyridin-3-yl)-N-(1-(2-methyl-3-(trifluoromethyl)phenyl)ethyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide can be synthesized in the same manner as steps 1 to 3 of Example 27 using 1-(5-bromopyridin-3-yl)-6-oxo-1,6-dihydropyridazine-3-carboxylic acid (3-1) and (1R)-1-[2-methyl-3-(trifluoromethyl)phenyl]ethanamine hydrogen chloride (A-3).

[0468] Example 31: (R)-1-(5-(1-(1,1-deoxidothietan-3-yl)-1H-1,2,3-triazol-5-yl)pyridin-3-yl)-N-(1-(2-(trideuteriomethyl)-3-(trifluoromethyl)phenyl)ethyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide

[0469]

[0470] (R)-1-(5-(1-(1,1-Dioxidothietan-3-yl)-1H-1,2,3-triazol-5-yl)pyridin-3-yl)-N-(1-(2-(trideuteriomethyl)-3-(trifluoromethyl)phenyl)ethyl)-6-oxo-1,6-dihydropyridazine-3-carboxamide can be synthesized in the same manner as steps 1 to 3 of Example 27 using 1-(5-bromopyridin-3-yl)-6-oxo-1,6-dihydropyridazine-3-carboxylic acid (3-1) and (1R)-1-[2-trideuterio-3-(trifluoromethyl)phenyl]ethanamine (A-5, HCl salt).

[0471] Example 32: 1-[5-[3-(3,3-difluorocyclobutyl)triazol-4-yl]-3-pyridyl]-N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethyl]-6-oxo-pyridine-3-carboxamide

[0472]

[0473] To a solution of 1-(5-bromopyridin-3-yl)-N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethyl]-6-oxo-pyridine-3-carboxamide (D-2,100 mg, 208.21 μmol) and 1-(3,3-difluorocyclobutyl)triazole (T-6,132.54 mg, 832.85 μmol) in DMF (6 mL) were added Pd(OAc)2 (18.70 mg, 83.29 μmol), K2CO3 (57.55 mg, 416.43 μmol), and XPhos (39.70 mg, 83.29 μmol), and then the mixture was degassed, purged with N2 three times, and stirred at 110 °C for 16 h under N2 atmosphere. It was stirred. The reaction mixture was filtered, and the filtrate was diluted with EtOAc (10 mL X 3). The combined organic layers were washed with brine (5 mL X 3), dried over anhydrous Na2SO4, and concentrated under reduced pressure to obtain a crude product, which was purified by preparative HPLC (column: F-Prepulite XP tC 18 40*200mm*7um; mobile phase: [water (NH3H2O+NH4HCO3)-MeCN]; gradient: 12%-52% B for 20 min). MeCN was removed by concentration under reduced pressure, and the residual solvent was removed by lyophilization to obtain the compound of Example 32 (3.1 mg, 19.86% yield, 100% purity) as a white solid. 1H NMR (400 MHz, CDCl3) δ ppm 8.77 (d,J= 2.4 Hz, 1H), 8.69 (d,J= 2.0 Hz, 1H), 8.18 (d,J= 2.4 Hz, 1H), 7.89 (t,J= 2.0 Hz, 1H), 7.86 (s, 1H), 7.73 (dd,J= 2.8, 9.6 Hz, 1H), 7.49 (t,J= 7.2 Hz, 1H), 7.44 (t,J= 7.2 Hz, 1H), 7.22 - 7.17 (m, 1H), 6.72 (d,J= 9.6 Hz, 1H), 6.40 (br d,J=8.0 Hz, 1H), 5.46 (t,J= 7.6 Hz, 1H), 4.84 (dt,J= 4.4, 8.0 Hz, 1H), 3.59 - 3.47 (m, 2H), 3.29 - 3.19 (m, 2H), 2.01 (t,J= 18.8 Hz, 3H), 1.64 (d,J= 7.2 Hz, 3H);MS (ESI) m / z = 559.2 [M+H] + .

[0474] Examples 33 to 51

[0475] The compounds in Table 2 below were prepared through a one-step Heck coupling using intermediates corresponding to the structure of the target compound based on the manufacturing example in the same manner as Example 32.

[0476]

[0477]

[0478]

[0479]

[0480]

[0481]

[0482]

[0483] Example 52: (R)-N-(1-(3-(1,1-difluoroethyl)-2-fluorophenyl)ethyl)-1-(5-(1-methyl-1H-1,2,3-triazol-5-yl)pyridin-3-yl)-6-oxo-5-(trifluoromethyl)-1,6-dihydropyridazine-3-carboxamide

[0484]

[0485] The compound of Example 52 (32.8 mg, 43.51% yield, 99.86% purity) was obtained as a yellow oil in the same manner as in step 3 of Preparation Example 17 using 1-[5-(3-methyltriazol-4-yl)-3-pyridyl]-6-oxo-5-(trifluoromethyl)pyridazine-3-carboxylic acid (5-2) and (1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethanamine (A-1). 1 H NMR (400 MHz, DMSO-d6) δ ppm 9.23 (d, J = 8.0 Hz, 1H), 9.10 (d, J = 2.4 Hz, 1H), 8.97 (d, J = 2.0 Hz, 1H), 8.50 (t, J = 2.0 Hz, 1H), 8.25 (s, 1H), 8.14 (s, 1H), 7.65 (t, J = 7.2 Hz, 1H), 7.47 (t, J = 6.8 Hz, 1H), 7.31 - 7.26 (m, 1H), 5.44 (quin, J = 7.2 Hz, 1H), 4.17 (s, 3H), 2.01 (t, J = 19.2 Hz, 3H), 1.51 (d, J = 7.2 Hz, 3H); MS (ESI) m / z = 552.6 [M+H] + .

[0486] Examples 53 to 61

[0487] The compounds in Table 3 below were prepared through amide coupling using intermediates corresponding to the structure of the target compound based on the manufacturing example in the same manner as Example 52.

[0488]

[0489]

[0490]

[0491]

[0492] Examples 62 and 63: 1-[5-[3-[(1S)-2,2-difluorocyclopropyl]triazol-4-yl]-3-pyridyl]-N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethyl]-6-oxo-pyridazine-3-carboxamide and 1-[5-[3-[(1R)-2,2-difluorocyclopropyl]triazol-4-yl]-3-pyridyl]-N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethyl]-6-oxo-pyridazine-3-carboxamide

[0493]

[0494] Step 1: 1-[5-[3-(2,2-difluorocyclopropyl)triazol-4-yl]-3-pyridyl]-N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethyl]-6-oxo-pyridazine-3-carboxamide

[0495] 1-(5-Bromopyridin-3-yl)-N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethyl]-6-oxo-pyridazine-3-carboxamide (D-3) and 1-(2,2-difluorocyclopropyl)triazole (T-5) were used in the same manner as in Example 14 to obtain 1-[5-[3-(2,2-difluorocyclopropyl)triazol-4-yl]-3-pyridyl]-N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethyl]-6-oxo-pyridazine-3-carboxamide (20 mg, 7.06% yield, 100% purity) as an off-white solid. MS (ESI) m / z = 546.2 [M+H] + .

[0496] Step 2: 1-[5-[3-[(1S)-2,2-difluorocyclopropyl]triazol-4-yl]-3-pyridyl]-N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethyl]-6-oxo-pyridazine-3-carboxamide and 1-[5-[3-[(1R)-2,2-difluorocyclopropyl]triazol-4-yl]-3-pyridyl]-N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethyl]-6-oxo-pyridazine-3-carboxamide

[0497] 1-[5-[3-(2,2-difluorocyclopropyl)triazol-4-yl]-3-pyridyl]-N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethyl]-6-oxo-pyridazine-3-carboxamide (20 mg, 36.67 μmol) was further purified using SFC separation (column: Daicel Chiralpak IBN 250mm*30mm*10um; mobile phase: [CO2-EtOH (0.1% NH3H2O)]; B%: 45%, isocratic elution mode). Two peaks were identified by SFC analysis. The compound identified in the first peak (peak 1, Rt = 1.434 min) was concentrated under reduced pressure. Afterwards, MeCN (5 mL) and H2O (25 mL) were added, and the solvent was removed by lyophilization to obtain 1-[5-[3-[(1S)-2,2-difluorocyclopropyl]triazol-4-yl]-3-pyridyl]-N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethyl]-6-oxo-pyridazine-3-carboxamide (Example 62, 6.1 mg, 30.50% yield, 100% purity, enantiomeric excess 99.90%) as a white solid. The absolute stereochemistry of [2,2-difluorocyclopropyl]triazol-4-yl was arbitrarily chosen as S. 1H NMR (400 MHz, DMSO-d6) δ ppm 9.15 (d, J = 2.4 Hz, 1H), 9.04 (d, J = 8.0 Hz, 1H), 9.00 (d, J = 2.0 Hz, 1H), 8.58 (t, J = 2.0 Hz, 1H), 8.26 (s, 1H), 7.93 (d, J = 9.6 Hz, 1H), 7.63 (t, J = 7.2 Hz, 1H), 7.46 (t, J = 6.8 Hz, 1H), 7.27 (t, J = 7.6 Hz, 1H), 7.23 (d, J = 9.6 Hz, 1H), 5.42 (quin, J = 7.6 Hz, 1H), 5.16 - 5.05 (m, 1H), 2.63 - 2.56 (m, 1H), 2.46 - 2.40 (m, 1H), 2.06 - 1.96 (m, 3H), 1.49 (d, J = 7.2 Hz, 3H); MS (ESI) m / z = 546.3 [M+H] + . The compound identified in the second peak (peak 2, Rt = 1.661 min) was concentrated under reduced pressure. Then, MeCN (5 mL) and H2O (25 mL) were added, and the solvent was removed by lyophilization to obtain 1-[5-[3-[(1R)-2,2-difluorocyclopropyl]triazol-4-yl]-3-pyridyl]-N-[(1R)-1-[3-(1,1-difluoroethyl)-2-fluorophenyl]ethyl]-6-oxo-pyridazine-3-carboxamide (Example 63, 6.3 mg, 31.31% yield, 99.41% purity, 99.54% enantiomeric excess) as a white solid. The absolute stereochemistry of [2,2-difluorocyclopropyl]triazol-4-yl was arbitrarily chosen as R. 1H NMR (400 MHz, DMSO-d6) δ ppm 9.15 (d, J = 2.4 Hz, 1H), 9.04 (d, J = 8.0 Hz, 1H), 9.00 (d, J = 2.0 Hz, 1H), 8.58 (t, J = 2.0 Hz, 1H), 8.26 (s, 1H), 7.93 (d, J = 9.6 Hz, 1H), 7.63 (t, J = 7.2 Hz, 1H), 7.46 (t, J = 6.8 Hz, 1H), 7.27 (t, J = 7.6 Hz, 1H), 7.23 (d, J = 9.6 Hz, 1H), 5.42 (quin, J = 7.6 Hz, 1H), 5.16 - 5.05 (m, 1H), 2.63 - 2.56 (m, 1H), 2.46 - 2.40 (m, 1H), 2.06 - 1.96 (m, 3H), 1.49 (d, J = 7.2 Hz, 3H); MS (ESI) m / z = 546.3 [M+H] + .

[0498] Examples 64 to 71

[0499] The compounds in Table 4 below were prepared through CH activation and SFC separation using intermediates corresponding to the structures of the target compounds based on the manufacturing examples in the same manner as in Examples 62 and 63. In Table 4 below, the absolute stereochemistry of [2,2-difluorocyclopropyl]triazol-4-yl was arbitrarily selected as S or R.

[0500]

[0501]

[0502]

[0503] Experimental Example 1: SOS-catalyzed nucleotide exchange assay

[0504] SOS1 activation is KRAS G12D SOS1-mediated GTP loading was measured by quantification of GTP-loaded KRAS.G12D was detected by HTRF (Homogeneous Time Resolved Fluorescence) using anti-GST terbium as donor (Cisbio, France) and a fluorescent GTP analogue (EDA-GTP-DY-647P1, [2' / 3'-O-(2-aminoethyl-carbamoyl)guanosine-5'-triphosphate labeled with DY-647P1 (Jena bioscience, Germany)]) as acceptor.

[0505] The assay buffer consisted of 10 mM HEPES pH 7.4, 150 mM NaCl, 5 mM MgCl2, 1 mM DTT, 0.05% BSA Fraction V pH 7.0 (Sigma), and 0.0025% (v / v) Igepal (Sigma). KRAS G12D The working solution contained an equal volume of GST-KRAS in assay buffer. G12D (100 nM) and anti-GST-terbium (2 nM; Cisbio, France) were added and incubated on ice for 10 min. SOS1 cat The working solution is an equal volume of HIS-SOS1 in assay buffer. cat (160 nM; Cytoskeleton, US) and EDA-GTP-DY-647P1 (200 nM; Jena bioscience, Germany) were added to prepare the control solution. The control solution was prepared by adding equal volumes of EDA-GTP-DY-647P1 and assay buffer for normalization control to exclude GEF-independent GTP loading and background signal. SOS1 was added to each well for reaction induction. cat 5 uL of working solution or control solution was injected. 5 uL of compounds with concentrations ranging from 128 pM to 10 μM were injected into SOS1 cat It was added to the wells containing the working solution and incubated at room temperature for 10 minutes to allow binding to SOS1.

[0506] 10 uL of KRAS G12DThe solution was added to each well and incubated with shaking at room temperature for 30 minutes. HTRF was measured using a Thermo Varioskan (excitation 334 nm, emission 665 nm). The IC of each inhibitor 50 was analyzed using GraphPad Prism.

[0507] The SOS1 inhibitory activity of the example compound measured by the above method was evaluated based on the following criteria, and the results are shown in Table 5.

[0508]

[0509] As shown in Table 5 below, the compounds of the present invention exhibited excellent SOS1 inhibitory activity.

[0510]

[0511]

[0512] Experimental Example 2: KRAS G12C Protein-protein interaction inhibition assay of SOS1 (KRAS) G12C ::SOS1 protein-protein interaction (PPI) inhibition assay)

[0513] SOS1 and KRAS G12C Protein-protein interactions were detected using the KRAS G12C / SOS1 binding kit (64KRASG12PEH, CISBIO, FRANCE) from CISBIO according to the manufacturer's assay method. Briefly, the inhibitor sample was diluted 7 times by 1 / 3 with a maximum concentration of 3 μM, and a total of 8 concentrations of samples were prepared. 2 μl of the inhibitor sample prepared for each concentration and 5 μl of Tag1-tagged KRAS diluted with the dilution solution provided by the manufacturer were added to a 96-well low-volume white plate dedicated to HTRF (Homogeneous Time-Resolved Fluorescence) (66PL96005, CISBIO, FRANCE). G12CMix the protein and GTP mixture solution, 5 μl of Tag2-labeled SOS1 protein, and 8 μl of anti-Tag1 KRAS mixed in the same ratio. G12C XL665 (HTRF acceptor) labeled antibody and anti-Tag2 SOS1 Terbium cryptate (HTRF donor) labeled antibody dilution solutions were added and incubated at room temperature for 2 hours. Afterwards, the HTRF signal generated from the antibody was measured using a Thermo Varioskan multipurpose plate signal detector, and the result was calculated as the ratio of the signals emitted at 665 nm and 620 nm, as shown below.

[0514] Result = 10 4 X 665 nm signal / 620 nm signal

[0515] IC of each inhibitor 50 The values ​​were analyzed using GraphPad Prism 9.0 software using the results of three replicate experiments.

[0516] SOS1 and KRAS of the example compounds measured by the above method G12C The degree of inhibition of liver cross-linking was evaluated based on the following criteria, and the results are shown in Table 6.

[0517]

[0518]

[0519]

[0520]

[0521] From the results in Table 6 below, the compounds of the present invention inhibit SOS1 and KRAS G12C It was confirmed that the cross-linking between the two was effectively suppressed.

[0522] Experimental Example 3: Confirmation of Pregnane X Receptor (PXR) Activation in a Reporter Assay System

[0523] The main mechanism of cytochrome P450 induction is generally through increased gene transcription, which occurs through nuclear receptor activation. PXR has been identified as the major nuclear receptor mediating drug-induced expression of CYP3A4 (Bertilsson G, et al., Proc Natl Acad Sci USA 95 12208-12213). Based on this CYP3A4 induction pathway, cellular PXR receptor gene analysis has been used to screen novel molecular compounds (NCEs) for drug interactions (Luo G, et al., Drug Metab Dispos. 2002 Jul, 30 (7): 795-804). To evaluate the CYP induction potential of the compounds disclosed herein, a PXR activation assay was performed.

[0524] To assess the potential of test substances to induce human PXR activation, the Human PXR Reporter Assay System provided by Indigo Biosciences (USA) was used. Briefly, this assay is based on cells genetically engineered to express a luciferase reporter gene linked to human PXR and a GAL4 DNA-binding sequence, and when a test substance activates PXR, transcription of the luciferase gene is induced, allowing for the quantitative assessment of PXR activation through the amount of light produced.

[0525] PXR reporter cells were seeded in 384-well plates and cultured for 4–6 hours in a 37 °C incubator supplied with 5% CO2. Each well was then treated with the test substance, positive control (rifampicin), or solvent control (final DMSO concentration 0.4%), and cultured for 22–24 hours under the same conditions. The test substances were dissolved in DMSO and diluted in the medium to final concentrations of 0.16, 0.5, 1.58, 5.01, 15.8, and 50 μM, respectively, and all concentrations were repeated twice (n = 2). The positive control, rifampicin, was treated at concentrations of 0.032, 0.1, 0.32, 1.0, 3.2, and 10 μM. The final DMSO concentration in all experimental groups was maintained at 0.4%. After the culture was completed, the medium was removed, and cell viability was measured by fluorescence using the Live Cell Multiplex Assay Kit from Indigo Biosciences. Luciferase activity was then measured using the Luciferase Detection Reagent included in the PXR Assay Kit, and the amount of luminescence was quantified using a plate reader.

[0526] Luciferase luminescence values ​​were normalized to cell viability, and the activation level at each test substance concentration was calculated as fold induction compared to the solvent control. The dose-response curve for the test substance was log-transformed and then analyzed by nonlinear regression to determine the EC 50 (concentration that induces 50% of the response) and E max (maximum response value) was derived (using GraphPad Prism). However, if the curve is irregular or proper fitting is not possible, EC 50 The output was omitted.

[0527] To compare activities between test substances and ensure consistency between experiments, the results were also analyzed as a relative activation percentage (%) for 10 μM rifampicin, but in this patent, only the fold induction value at a concentration of 15.8 μM is described in Table 7 for convenience of comparison.

[0528]

[0529]

[0530] From the results in Table 7 above, it was confirmed that the compounds of the present invention generally showed an effect less than the threshold value on PXR activation.

[0531] Experimental Example 4: Confirmation of Time-Dependent Inhibition of Cytochrome P450

[0532] Time-dependent inhibition (TDI) is an assay that evaluates the time-dependent inhibition potential of parent drug and metabolites in liver microsomes. The result is converted to the compound's metabolite using NADPH, and the IC 50 , and IC without NADPH 50 The results of measuring the change were then diluted with a fresh experimental buffer containing NADPH and CYP3A4 midazolam or testosterone, and the degree of hydroxylation inhibition was measured by LC / MS / MS. IC 50 If the change is greater than 1.5-fold, time-dependent inhibition, where 30 minutes of pre-incubation results in an increase in efficacy, is considered positive.

[0533] Time-dependent inhibition measurements of cytochrome P450 were performed using human liver microsomes at Wuxi University.

[0534] Prepare test compound and positive control working solutions (100X) in organic solvent, remove microsomes from the -80.0°C freezer, and thaw. Prepare microsome solution and add 147.5 μL to all incubation plate wells. Prepare auxiliary solution and enzyme marker substrate solution. Add 2.50 μL of test compound and positive control working solutions to the corresponding wells, and add 2.50 μL of 1:1 DMSO / MeOH to the NIC wells. Preheat the plate at 37.0°C for approximately 10 minutes. Add 50.0 μL PB to the preincubation wells, add 50.0 μL cofactor, mix, and preincubate in a 37.0°C water bath for 30 minutes. After that, 50.0 μL of the cofactor and substrate mixture was added to the incubation well, and 50.0 μL of the substrate was added and mixed, and incubated in a 37.0°C water bath for 3 minutes for the CYP3A (Midazolm) reaction and 10 minutes for the CYP3A (Testosterone) reaction. When the set time was reached, 250 μL of the chilled IS fortified stop solution was added to all wells to stop the reaction. Centrifuge the incubation plate at 4000 rpm for 20 minutes, take 180 μL of the supernatant, mix it with 180 μL of HPLC water, and shake it for 10 minutes. This completes the sample preparation for injection.

[0535] The test compounds were measured at 0, 0.05, 0.165, 0.5, 1.65, 5, 16.5, and 50 μM, and Verapamil was used as a positive control. The percentage of the control group and the test compound concentration were plotted using XL fit, and nonlinear regression analysis of the data was performed. IC 50 Values ​​were determined using a 3- or 4-parameter logistic equation.

[0536] IC of the example compound measured by the above method 50When the shift ratio was 1.5 or higher, it was classified into the following grades, considering that it was positive for time-dependent inhibition. The results are shown in Table 8.

[0537]

[0538]

[0539] From the results in Table 8 above, it was confirmed that the compound of the present invention was negative for time-dependent inhibition of CYP3A4 Midazolam and Testosterone.

[0540] Experimental Example 5: Confirmation of CYP3A4 induction using human hepatocytes

[0541] Whether the compound of the invention increases the expression of cytochrome P450 enzymes in vivo was evaluated using cryopreserved human hepatocytes (Cyprotex).

[0542] 0.072 Х 10 cryopreserved human hepatocytes from a single donor 6 Cells were seeded into 96-well collagen-coated plates at a density of 10 cells / well (final volume 0.1 mL per well) and cultured at 37 °C, 95% humidity, and 25% CO2 to allow attachment. After at least 4 h, the medium was replaced with pre-warmed serum-free Williams' E medium (containing 100 IU / mL penicillin, 100 μg / mL streptomycin, 10 μg / mL insulin, 2 mM L-glutamine, and 0.1 μM hydrocortisone) and incubated for approximately 24 h, after which it was replaced with assay medium containing the test compound. Test compounds were treated in triplicate at each concentration for 72 h, and the medium was changed every 24 h.

[0543] For mRNA measurement, after 72 hours of exposure, all media from each well was removed, washed once with 200 μL of pre-warmed PBS, and 100 μL of lysis solution (10 mM Tris containing 300 mM sodium chloride, 0.1% IGEPAL, and 1:1000 RNasin® Ribonuclease inhibitor) was added to each well to lyse the cells. Reverse transcription was then performed, and quantitative PCR analysis was performed on the generated cDNA using gene-specific primer probe sets for CYP1A2, CYP2B6, CYP2C8, CYP2C9, and CYP3A4 target cDNAs and an endogenous control. Samples were analyzed using an Applied Biosystems QuantStudio™ 7 Real-Time PCR system.

[0544] Test compounds were measured at concentrations of 0.2, 0.5, 2, 5, 20, and 50 μM, and rifampicin was used as a positive control. Control cultures treated with only solvent (typically 0.1% DMSO in the experimental medium) instead of test compounds were also included.

[0545] Data analysis was performed to determine the critical cycle (C) of the target gene compared to the endogenous control. T ) based on the data to measure relative fold mRNA expression and 2 -ΔΔCT Normalized to the solvent control using the method. Nonlinear regression analysis of the log dose-response curve (using a four-parameter sigmoidal model) yielded EC 50 and E max The values ​​were determined and the results are shown in Table 9 and Figure 1.

[0546]

[0547] From the experimental results in Table 9 and Figure 1, it was confirmed that the compounds of Examples 1 and 56 showed significantly lower induction folds of less than half that of the positive control group, and thus could be used without concern for clinically serious drug interactions.

[0548] Experimental Example 6: Evaluation of tumor growth inhibition efficacy according to compound administration in the H358 tumor subcutaneous transplantation model.

[0549] In a model where human lung cancer cells, H358 cells, were subcutaneously transplanted into BALB / c nude mice, the tumor growth inhibitory effect of combined administration of the example compound and sotorasib was confirmed.

[0550] H358 cells (Korean Cell Line Bank, No. 25807) were cultured in RPMI medium supplemented with 10% heat-inactivated fetal bovine serum, antibiotics, and antimycotics at 37°C and 5% CO2 according to the supplier's culture method. Tumor cells were passaged 2–3 times a week to maintain 70–80% confluency using trypsin-EDTA treatment, and cells in the exponential growth phase before transplantation were harvested and counted.

[0551] Tumor cells were cultured in 0.1 mL of PBS / Matrigel 1:1 mixture for tumor growth. H358 cells (5x10 6 The tumors were subcutaneously implanted into the upper right flank of each mouse to include cells. Approximately one week after tumor implantation, the average tumor size was approximately 226 mm. 3When the target was reached, group separation was performed so that the average tumor size of each group was similar. Tumor size was measured while repeatedly administering the test substance according to the test design in Table 10 below. Vehicle (0.5% medium-chain triglyceride (MCT)) was administered to the negative control group (G1), sotorasib, a clinically approved KRAS inhibitor, was administered as the positive control group (G2), and MRTX0902, an SOS1 inhibitor currently in clinical trials, was administered in combination with sotorasib as the comparison group (G3). The tumor inhibition rate was measured 20 days after group assignment (D20), and administration was carried out for 21 days, with the day of group assignment being designated as Day 0. The measurement results are shown in Table 11 and Fig. 2.

[0552]

[0553]

[0554]

[0555] As can be seen from the results in Table 11 and Fig. 2, the example compounds, when co-administered with sotorasib, exhibited significantly improved tumor inhibition effects compared to sotorasib alone (G2). Furthermore, when compared to the group co-administered with MRTX0902 and sotorasib (G3), it was confirmed that the example compounds exhibited an equally excellent tumor inhibition effect or better even at a dose less than half that of MRTX0902.

Claims

1. A compound represented by the following chemical formula I, or a solvate, stereoisomer or pharmaceutically acceptable salt thereof: [Chemical Formula I] In the above chemical formula I, Z 1 is N or CH, A 1 is CF3, CH3-CF2- or CH2F-CF2-, A 2 is halogen; or C1-C6 alkyl optionally substituted with one or more deuterium, R 1 is H; or C1-C3 alkyl optionally substituted with halogen, R 2 is a straight or branched C1-C6 alkyl optionally substituted with one or more deuterium or halogen; a C3-C7 cycloalkyl optionally substituted with one or more deuterium, halogen or C1-C3 alkyl; or a 3- to 7-membered heterocyclyl comprising a heteroatom or heteroatom group selected from N, O, S and SO2.

2. In paragraph 1, A 2 A compound, or a solvate, stereoisomer or pharmaceutically acceptable salt thereof, wherein the compound is F, CH3 or CD3.

3. In paragraph 1, A compound, or a solvate, stereoisomer or pharmaceutically acceptable salt thereof, selected from the following structures: , and .

4. In paragraph 1, R 2 A compound, or a solvate, stereoisomer or pharmaceutically acceptable salt thereof, wherein the compound is a straight or branched C1-C3 alkyl optionally substituted with one or more deuterium atoms; a C3-C5 cycloalkyl optionally substituted with one or more halogens or methyl; or a heterocyclyl selected from oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, 1,1-dioxidothietanyl and 1,1-dioxidotetrahydrothiophenyl.

5. In paragraph 4, R 2 is CH3, CD3, -CH2CH3, -CH(CH3)2, cyclopropyl, cyclobutyl, , , or A compound, or a solvate, stereoisomer or pharmaceutically acceptable salt thereof.

6. In paragraph 1, R 2 A compound, or a solvate, stereoisomer or pharmaceutically acceptable salt thereof, wherein the compound is C3-C7 cycloalkyl optionally substituted with one or more halogens or C1-C3 alkyl; or a 3- to 7-membered heterocyclyl comprising a heteroatom or heteroatom group selected from N, O, S and SO2.

7. In paragraph 1, R 2 A compound, or a solvate, stereoisomer or pharmaceutically acceptable salt thereof, which is a straight or branched C1-C6 alkyl optionally substituted with one or more deuterium atoms.

8. In paragraph 1, R 1 A compound, or a solvate, stereoisomer or pharmaceutically acceptable salt thereof, wherein said compound is H, CH3 or CF3.

9. In paragraph 1, A compound represented by the following chemical formula IC, or a solvate, stereoisomer or pharmaceutically acceptable salt thereof: [Chemical formula IC] In the above chemical formula IC, Z 1 is N or CH, R 1 is H; or C1-C3 alkyl optionally substituted with halogen, R 2 is a straight or branched chain C1-C6 alkyl optionally substituted with one or more deuterium; or a C3-C7 cycloalkyl optionally substituted with one or more deuterium or halogen.

10. In paragraph 9, R 1 is C1-C3 alkyl substituted with H or halogen, R 2 A compound, or a solvate, stereoisomer or pharmaceutically acceptable salt thereof, wherein the compound is a straight or branched C1-C3 alkyl optionally substituted with one or more deuteriums; or a C3-C5 cycloalkyl optionally substituted with one or more deuteriums or halogens.

11. In paragraph 10, R 1 is H, R 2 is CH3, CD3 or cyclopropyl, A compound, or a solvate, stereoisomer or pharmaceutically acceptable salt thereof.

12. In paragraph 1, A compound selected from the group consisting of: or a solvate, stereoisomer or pharmaceutically acceptable salt thereof; 13. A compound selected from the group consisting of: or a solvate, stereoisomer or pharmaceutically acceptable salt thereof:

14. A pharmaceutical composition comprising a compound of any one of claims 1 to 13, or a solvate, stereoisomer or pharmaceutically acceptable salt thereof, as an active ingredient.

15. In paragraph 14, A pharmaceutical composition for the prevention or treatment of SOS1-mediated diseases.

16. In paragraph 15, A pharmaceutical composition wherein the above SOS1-mediated disease is cancer or RAS disease.

17. In paragraph 16, The above cancer is selected from the group consisting of pancreatic cancer, lung cancer, colorectal cancer, cholangiocarcinoma, multiple myeloma, melanoma, uterine cancer, cervical cancer, endometrial cancer, thyroid cancer, chronic lymphocytic leukemia, acute myeloid leukemia, bladder cancer, urothelial cancer, stomach cancer, head and neck squamous cell carcinoma, diffuse large B-cell lymphoma, esophageal cancer, hepatocellular cancer, breast cancer, ovarian cancer, prostate cancer, glioblastoma, renal cancer, and sarcoma. A pharmaceutical composition, wherein the RAS disease is selected from the group consisting of Neurofibromatosis type 1, Noonan Syndrome, Leopard Syndrome, Capillary Malformation-Arteriovenous Malformation Syndrome, Costello Syndrome, Cardio-Facio-Cutaneous Syndrome, Legius Syndrome, and Hereditary gingival fibromatosis.

18. A pharmaceutical composition for preventing or treating cancer, comprising a compound of any one of claims 1 to 13, or a solvate, stereoisomer, or pharmaceutically acceptable salt thereof; and a targeted anticancer agent as an active ingredient.

19. In paragraph 18, A pharmaceutical composition wherein the targeted anticancer agent acts on one or more targets selected from Bcl-2, EGFR, KRAS, MEK, and RAF.

20. In paragraph 19, A pharmaceutical composition wherein the targeted anticancer agent is selected from venetoclax, zongitinib, osimertinib, lazertinib, cetuximab, sotorasib, adagrasib, dibarasib, MRTX1133, trametinib, and abutometinib.

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