Novel NLRP3 inhibitor compound

Abstract

The present invention relates to a heteroaryl derivative compound and a use thereof. A heteroaryl derivative according to the present invention exhibits excellent inhibitory activity against NLRP3, and thus can be effectively used for the treatment or prevention of NLRP3-related diseases.

Description

Novel NLRP3 inhibitor compounds

[0001] The present invention relates to heteroaryl derivative compounds and pharmaceutical uses thereof, and more particularly, to heteroaryl derivative compounds having NLRP3 inhibitory activity and pharmaceutical uses thereof.

[0002] NLRP3 (NOD-, LRR-, and pyrin domain-containing protein 3) is an intracellular sensor capable of detecting a wide range of microbial motifs, endogenous danger signals, and environmental stimuli. The NLRP3 protein is structurally composed of PYD (amino-terminal pyrin domain), NACHT (domain present int NAIP, CIITA, HET-E, and TP1), and LRR (leucine-rich repeat) domains, of which the NACHT domain possesses ATPase activity essential for NLRP3 self-assembly and function.

[0003] When NLRP3 proteins are stimulated within a cell, NLRP3 proteins interact with each other through the NACHT domains within NLRP3 to form oligomers, recruiting ASC protein to form ASC specks, and subsequently recruiting inactive Caspase-1 protein. As a result, a large protein complex called NLRP3 inflammasome is formed, consisting of NLRP3 protein (sensor function), ASC protein (adapter function), and Caspase 1 protein (effector function). Then, Caspase-1 is self-cleaved from inactive Caspase-1, and activated Caspase-1 releases potent proinflammatory cytokines IL-1β and IL-18 outside the cell, and pyroptosis cell death mediated by GSDMD (Gasdermin D) is induced (reviewed in [Swanson et al. Nature Reviews Immunology 19.8 (2019): 477-489.]). As a result, it has been reported that various NLRP3-related diseases, such as Alzheimer's disease, multiple sclerosis, atherosclerosis, and inflammatory bowel disease, are induced or worsened.

[0004] Accordingly, the NLRP3 protein has been considered as a drug target for NLRP3-related diseases, and compounds with various structures, such as MCC950, CY-09, Oridonin, Tranilast, MNS, OLT1177 (dapansutrile), BAY 11-7082, BOT-4-one, Parthenolide, and INF39, have been studied and developed as NLRP3 inflammasome inhibitors.

[0005] However, small molecule compounds developed as NLRP3 protein inhibitors to date have limitations in inhibiting the characteristic enzyme activity in the target protein due to the structural characteristics of the NLRP3 protein, and clinical problems such as off-target side effects, short blood half-life, and hepatotoxicity have been reported during the development process (literature [Mangan, Matthew SJ, et al. Nature reviews Drug discovery 17.8 (2018): 588-606.]).

[0006] Therefore, the development of novel small-molecule compounds is required to develop treatments for NLRP3-related diseases.

[0007] One object of the present invention is to provide a novel compound having NLRP3 inhibitory activity, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0008] Another object of the present invention is to provide a method for producing a novel compound having NLRP3 inhibitory activity.

[0009] Another object of the present invention is to provide a pharmaceutical use of the novel compound, and specifically, to provide a pharmaceutical composition for treating or preventing a NLRP3-related disease, which comprises the compound as an active ingredient.

[0010] Another object of the present invention is to provide a method for treating or preventing an NLRP3-related disease using the compound or a method for treating or preventing an NLRP3-related disease comprising a step of administering the compound.

[0011] In order to achieve the above purpose, the inventors of the present invention completed the present invention by newly synthesizing a heteroaryl derivative compound represented by the chemical formula 1 mentioned below and confirming the NLRP3 inhibitory activity of the compounds as a result of research efforts.

[0012] NLRP3 inhibitory activity compounds

[0013] The present invention provides a compound represented by the following chemical formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0014] [Chemical Formula 1]

[0015]

[0016] In the above chemical formula 1,

[0017] R1 is -H, -C 1-4 Alkyl, -C 1-4 Haloalkyl, or -halo;

[0018] X1 to X3 are each independently CR X or N;

[0019] R X is -H, -C 1-4 Alkyl, -C 1-4 Haloalkyl, or -halo;

[0020] R2 is -H, -C 1-4 Alkyl, -OH, -OC 1-4 Alkyl, -O-(3-6 membered cycloalkyl), or -halo;

[0021] Z1 to Z5 are each independently CR Z or N;

[0022] R Z is -H, -C 1-4 Alkyl, -C 1-4 Haloalkyl, -OH, or -halo;

[0023] Y is -azetidine or -CH2-W, wherein at least one H of the azetidine ring is -C 1-3 may be substituted with alkyl;

[0024] W is -(4-7 membered heterocycloalkyl), wherein said -(4-7 membered heterocycloalkyl) contains at least one N or O atom, and at least one H of said -(4-7 membered heterocycloalkyl) ring is -C 1-3alkyl, -(3-6 membered cycloalkyl), -(4-7 membered heterocycloalkyl), -phenyl, or -(5-6 membered heteroaryl), wherein at least one H of the -phenyl or -(5-6 membered heteroaryl) ring is -C 1-3 Alkyl, -O-R1 or -NR1R2;

[0025] R1 and R2 are each independently -H, -C 1-3 Alkyl, -C(=O)H or -C(=O)-C 1-3 It's alkyl.

[0026] According to one specific example of the present invention,

[0027] R1 is -H;

[0028] X1 is CR X and;

[0029] X2 is CR X or N;

[0030] X3 is CR X and;

[0031] R X is -H, -C 1-4 Alkyl, -C 1-4 It can be haloalkyl, or -halo.

[0032] According to one specific example of the present invention,

[0033] R2 is -OC 1-4 It can be alkyl or -O-(3-6 membered cycloalkyl).

[0034] According to one specific example of the present invention,

[0035] Z1, Z2, Z4, or Z5 are each independently CR Z and;

[0036] Z3 is CR Z or N;

[0037] R Z is -H, -C 1-4 Alkyl, -C 1-4 It can be haloalkyl, or -halo.

[0038] According to one specific example of the present invention,

[0039] Y is -azetidine or -CH2-W, wherein at least one H of the azetidine ring is -C 1-3 may be substituted with alkyl;

[0040] W is , , , , , or and;

[0041] R A is -H, -C 1-3 which may be substituted with alkyl, -(3-6 membered cycloalkyl), or -phenyl, wherein one or more H of the -phenyl ring may be substituted with -NR1R2;

[0042] R1 and R2 are each independently -H, -C 1-3 Alkyl, or -C(=O)-C 1-3 It could be an alkyl.

[0043] In the present invention, "alkyl" may mean a straight or branched chain, cyclic or saturated hydrocarbon or a combination thereof, unless otherwise specified. For example, "C 1-6 "Alkyl" may mean alkyl having 1 to 6 carbon atoms. Acyclic alkyl may include, but is not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, isopentyl, and 2-methylpentyl, for example. As used herein, a residue obtained by removing one hydrogen atom from the "alkyl" is referred to as "alkylene." Cyclic alkyl may be used interchangeably with "cycloalkyl" as used herein, and may include, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or cyclooctyl, for example.

[0044] In the present invention, "alkenyl" and "alkynyl" may mean a straight or branched chain acyclic, cyclic or unsaturated hydrocarbon group combined therewith. For example, "C 2-6 "Alkenyl" may mean an unsaturated hydrocarbon having 2 to 6 carbon atoms with one or more double bonds, and "C 2-6 "Alkynyl" may mean an unsaturated hydrocarbon having 2 to 6 carbon atoms with one or more triple bonds.

[0045] In the present invention, "alkoxy" may mean an alkyl ether group -(R'-OR"), where R' is a single bond and C 1-6 may be selected from the group consisting of alkyl, and R" is C 1-6 It may be alkyl. Here, alkyl is as defined above. For example, "C 1-6 "Alkoxy" of C 1-6 Alkoxy containing alkyl, i.e., -(OC 1-6 alkyl) or -(C 1-6 Alkyl-OC 1-6 Alkoxy may mean, but is not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, or tert-butoxy.

[0046] In the present invention, “halo” may be F, Cl, Br, or I.

[0047] In the present invention, "haloalkyl" may mean a straight or branched chain alkyl (hydrocarbon) having carbon atoms substituted with one or more halo groups as defined herein. Examples of such haloalkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, or tert-butyl, each independently substituted with one or more halogens, such as F, Cl, Br, or I.

[0048] In the present invention, "hydroxyalkyl" may mean a straight or branched chain alkyl (hydrocarbon) having a carbon atom substituted with hydroxy (OH). Examples of such hydroxyalkyl include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, or tert-butyl independently substituted with -OH.

[0049] In the present invention, "alkylamino" or "aminoalkyl" may mean -(NR'R"), wherein R' and R" are each independently hydrogen, C 1-6 alkyl, and N protecting groups (e.g., Boc), wherein the selected R' and R" may each be independently substituted or unsubstituted. In addition, "C 0-6 "Alkylamino" is amino (-NH2) or C that does not contain alkyl. 1-6 Amino containing alkyl, i.e., -NH(C 1-6 alkyl) or -N(C 1-6 alkyl)2, and may include, but is not limited to, dimethylamino, diethylamino, methylethylamino, methylpropylamino, or ethylpropylamino.

[0050] In the present invention, “cyanoalkyl” may mean a straight or branched chain alkyl (hydrocarbon) having a carbon atom substituted with cyano (CN).

[0051] In the present invention, "alkylsulfonyl" may mean -(R'-S(=O)2-R"), where R' is a single bond and C 1-6 may be selected from the group consisting of alkyl, and R" is hydroxy and C 1-6 It can be selected from the group consisting of alkyl. The selected R' and R" can be independently substituted or unsubstituted. In addition, "C 0-6 "Alkylsulfonyl" is a sulfonic acid group (-S(=O)2OH) or C that does not contain alkyl. 1-6 A sulfonyl group containing an alkyl group, i.e., -S(=O)2-(C 1-6 alkyl) or -(C 1-6 alkyl)-S(=O)2-(C 1-6 alkyl) and may include, but is not limited to, methylsulfonyl, (methylsulfonyl)methyl, (methylsulfonyl)ethyl, ethylsulfonyl, (ethylsulfonyl)methyl, and (ethylsulfonyl)ethyl.

[0052] In the present invention, "alkylcarbonyl" may mean -(R'-C(=O)-R"), where R' is a single bond and C 1-6 may be selected from the group consisting of alkyl, and R" is hydrogen and C 1-6 It can be selected from the group consisting of alkyl. The selected R' and R" can be independently substituted or unsubstituted. In addition, "C 0-6 "Alkylcarbonyl" is an aldehyde group (-C(=O)H) or C that does not contain alkyl. 1-6 A ketone group containing alkyl, i.e., -C(=O)-(C 1-6 alkyl) or -(C 1-6 alkyl)-C(=O)-(C 1-6 It can mean alkyl.

[0053] In the present invention, "cycloalkyl" may mean a hydrocarbon ring that does not contain a heteroatom (such as N, O, P, P(=O), or S) within the ring, and may be saturated or partially unsaturated. Here, if unsaturated, it may be referred to as a cycloalkene. Unless otherwise stated, a cycloalkyl may be a single ring or multiple rings such as a spiro ring, a bridged ring, or a fused ring.

[0054] In the present invention, "heterocycloalkyl" may mean a ring containing one or more selected from N, O, P, P(=O), and S within the ring, and may be saturated or partially unsaturated. Here, when unsaturated, it may be referred to as heterocycloalkene. Unless otherwise stated, a heterocycloalkyl may be a single ring or multiple rings such as a spiro ring, a bridged ring, or a fused ring. Additionally, "heterocycloalkyl having 3 to 12 atoms" may mean a heterocycloalkyl having 3 to 12 ring-forming atoms, and as an example, the heterocycloalkyl may be pyrrolidine, piperidine, imidazolidine, pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin, dioxolane, pyrimidine-2,4(1H,3H)-dione, 1,4-dioxane, morpholine, thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, piperazine, pyran, pyridone, 3-pyrroline, thiopyran, tetrahydrofuran, tetrahydrothiophene, quinuclidine, tropane, 2-azaspiro[3.3]heptane, It may include, but is not limited to, (1r,5s)-3-azabicyclo[3.2.1]octane, (1s,4s)-2-azabicyclo[2.2.2]octane, or (1r,4r)-2-oxa-5-azabicyclo[2.2.2]octane.

[0055] In the present invention, "heterobicycloalkyl" may mean multiple rings such as a spiro ring, a bridged ring or a fused ring containing at least one selected from N, O, P, P(=O), and S within the ring, and may be saturated or partially unsaturated. Here, when unsaturated, it may be referred to as heterobicycloalkene. Examples of heterobicycloalkyl may include, but are not limited to, quinuclidine, tropane, 2-azaspiro[3.3]heptane, (1r,5s)-3-azabicyclo[3.2.1]octane, (1s,4s)-2-azabicyclo[2.2.2]octane, or (1r,4r)-2-oxa-5-azabicyclo[2.2.2]octane.

[0056] In the present invention, "arene" may mean an aromatic hydrocarbon ring. The arene may be a monocyclic arene or a polycyclic arene. The number of ring-forming carbon atoms of the arene may be 5 to 30, 5 to 20, or 5 to 15. Examples of arenes include, but are not limited to, benzene, naphthalene, fluorene, anthracene, phenanthrene, bibenzene, terbenzene, quaternary benzene, quincbenzene, sexibenzene, triphenylene, pyrene, benzofluoranthene, chrysene, and the like. In the present specification, a residue obtained by removing one hydrogen atom from the "arene" is referred to as "aryl."

[0057] In the present invention, "heteroarene" may be a ring containing at least one of O, N, P, Si, and S as a heteroatom. The number of ring-forming atoms of the heteroarene may be 3 or more and 30 or less, 3 or more and 20 or less, or 3 or more and 15 or less. The heteroarene may be a monocyclic heteroarene or a polycyclic heteroarene. The polycyclic heteroarene may have, for example, a two-ring or three-ring structure. Examples of heteroarenes include thiophene, purine, pyrrole, pyrazole, imidazole, thiazole, oxazole, isothiazole, oxadiazole, triazole, pyridine, pyridin-2-one, pyridin-3-one, pyridin-4-one, bipyridine, triazine, acridyl, pyridazine, pyrazine, quinoline, quinazoline, quinoxaline, phenoxazine, phthalazine, pyrimidine, pyridopyrimidine, pyridopyrazine, pyrazinopyrazine, isoquinoline, indole, carbazole, imidazopyridazine, imidazopyridine, imidazopyrimidine, pyrazolopyrimidine, imidazopyrazine or Pyrazolopyridine, N-arylcarbazole, N-heteroarylcarbazole, N-alkylcarbazole, benzoxazole, benzimidazole, benzocarbazole, benzothiophene, dibenzothiophene, thienothiophene, benzofuran, phenanthroline, isoxazole, thiadiazole, benzothiazole, tetrazole, phenothiazine, dibenzosilole, and dibenzofuran, but are not limited thereto. In one embodiment of the present invention, the heteroarene may also include a bicyclic heterocycloarene comprising an arene ring fused to a heterocycloalkyl ring or a heteroarene fused to a cycloalkyl ring. As used herein, a residue obtained by removing one hydrogen atom from the above "heteroarene" is referred to as a "heteroaryl".

[0058] In the present invention, “hydroarene” or “hydroaryl” is an aromatic hydrocarbon ring in which at least one double bond is saturated.

[0059] In the present invention, "heterohydroarene" may mean a multi-ring (bi- to tetra-ring) containing 1 to 5 heteroatoms selected from N, O, and S as ring-forming atoms, and at least one of the multi-rings may be a saturated or partially unsaturated ring, and at least another may have an aromatic ring. In the present specification, a residue obtained by removing one hydrogen atom from the "heterohydroarene" is referred to as a "heterohydroaryl."

[0060] In the present invention, the "ring" may be a single ring or a multi-ring. The multi-ring may be a spiro ring, a bridged ring, or a fused ring.

[0061] In the present invention, "stereoisomer" means a compound having the same chemical formula or molecular formula but being sterically different. In the present specification, stereoisomers include optical isomers, enantiomers, diastereomers, cis / trans isomers, rotamers, and atropisomers, and each of these isomers, racemates, and mixtures thereof are also included in the scope of the present invention. For example, the compound represented by Chemical Formula 1 of the present invention may include the above stereoisomers of Chemical Formula 1 because the stereochemical structure is not specified. Unless otherwise stated, solid bonds connecting asymmetric carbon atoms A wedge-shaped solid line combination representing the absolute arrangement of the three-dimensional centers or wedge-shaped dotted line combination may include.

[0062] The compound represented by Chemical Formula 1 of the present invention may exist in the form of a "pharmaceutically acceptable salt." Accordingly, the category of the compound of the present invention includes a pharmaceutically acceptable salt of the compound represented by Chemical Formula 1. The term "pharmaceutically acceptable salt" of the present invention means any organic or inorganic acid addition salt of the compound, which has a relatively non-toxic and harmless effective effect in a patient at a concentration, and wherein the side effects caused by the salt do not reduce the beneficial effects of the compound represented by Chemical Formula 1.

[0063] In particular, the pharmaceutically acceptable salt may be an acid addition salt formed by a free acid. Here, the acid addition salt can be obtained from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid, etc.; non-toxic organic acids such as aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, hydroxyalkanoates and alkanedioates, aromatic acids, aliphatic and aromatic sulfonic acids, etc.; organic acids such as trifluoroacetic acid, acetate, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid, etc.

[0064] Such pharmaceutically acceptable salts may include sulfate, sulfite, nitrate, phosphate, pyrophosphate, chloride, bromide, iodide, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, benzoate, phthalate, benzenesulfonate, toluenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, glycolate, malate, tartrate, mandelate, and the like.

[0065] The above acid addition salt can be prepared by a conventional method, for example, by dissolving the derivative of chemical formula 1 in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile, etc., adding an organic acid or inorganic acid, filtering and drying the resulting precipitate, or by distilling the solvent and an excess acid under reduced pressure, drying, and crystallizing in an organic solvent.

[0066] In addition, the pharmaceutically acceptable salt may be a salt or metal salt obtained using a base. As an example of a metal salt, an alkali metal or alkaline earth metal salt can be obtained by dissolving a compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering out the undissolved compound salt, and evaporating and drying the filtrate. As an alkali metal salt, sodium, potassium, or calcium salts may be pharmaceutically suitable. In addition, a corresponding salt can be obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (e.g., silver nitrate), and can be prepared through a salt preparation method known in the art.

[0067] Uses of compounds with NLRP3 inhibitory activity

[0068] The present invention provides a use of a compound represented by the following chemical formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0069] [Chemical Formula 1]

[0070]

[0071] The above chemical formula 1 is as defined above.

[0072] The present invention provides uses of compounds, stereoisomers thereof, or pharmaceutically acceptable salts thereof.

[0073] The compound of the present invention, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof exhibits inhibitory activity against NLRP3.

[0074] According to one specific example of the present invention, the compound of the present invention exhibits excellent inhibitory activity against NLRP3, and thus can be usefully used for the treatment or prevention of NLRP3-related diseases.

[0075] In the present invention, the term "NLRP3-associated disease" refers to a disease or condition that can be mediated by an inflammatory pathway activated by the NLRP3 inflammasome, and can be treated, alleviated, delayed, inhibited, or prevented by degradation of the NLRP3 protein. The specific names and scope of NLRP3-associated diseases are known in the technical field to which the present invention pertains (e.g., literature [Swanson et al. Nature Reviews Immunology 19.8 (2019): 477-489.]).

[0076] For example, NLRP3-associated diseases include central nervous system diseases (e.g., Alzheimer's disease, multiple sclerosis, amyotrophic lateral sclerosis, or Parkinson's disease), metabolic disorders (e.g., type 1 diabetes, type 2 diabetes, hypertension, atherosclerosis, obesity, or gout), cardiovascular diseases (e.g., myocardial infarction, giant cell arteritis), respiratory diseases (e.g., asthma, COPD, silicosis, pulmonary diuretic fibrosis, allergic airway inflammation), liver diseases (e.g., nonalcoholic steatohepatitis [NASH], viral hepatitis, or cirrhosis), pancreatic diseases (e.g., acute pancreatitis or chronic pancreatitis), kidney diseases (e.g., nephropathy, acute kidney injury, or chronic kidney injury), intestinal diseases (e.g., inflammatory bowel diseases such as Crohn's disease or ulcerative colitis), skin diseases (e.g., psoriasis), musculoskeletal diseases (e.g., scleroderma), bone diseases (e.g., Osteoarthritis, osteoporosis, osteopetrosis), eye diseases (e.g., glaucoma, macular degeneration), inflammation after viral infections (e.g., inflammation after HIV, influenza, chikungunya, or COVID-19 virus infection), autoimmune diseases (e.g., rheumatoid arthritis, systemic lupus erythematosus, graft-versus-host disease [GVHD], autoimmune thyroiditis, autoimmune encephalitis), cancer or tumors (e.g., myelodysplastic syndrome, non-small cell lung cancer, metastatic lung cancer, gastric cancer, acute lymphoblastic leukemia [ALL], acute myeloid leukemia [AML], chronic myeloid leukemia [CML], promyelocytic leukemia, Langerhans cell histopathy, multiple myeloma), and other inflammatory diseases (e.g., contact hypersensitivity, traumatic brain injury, cryopyrin-associated periodic syndromes [CAPS]) Including, but not limited to, the following:

[0077] In one aspect, the present invention provides a pharmaceutical composition for treating or preventing an NLRP3-related disease, comprising a compound represented by Chemical Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0078] The pharmaceutical composition of the present invention may further include one or more active ingredients exhibiting the same or similar efficacy in addition to the compound represented by the chemical formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0079] The pharmaceutical composition of the present invention can be used for clinical administration and can be prepared so that it can be administered in various oral and parenteral dosage forms.

[0080] In addition, according to one specific example of the present invention, the present invention provides the use of a compound represented by the above chemical formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention or treatment of a NLRP3-related disease. The compound of the present invention for the manufacture of a medicament may be mixed with acceptable adjuvants, diluents, carriers, etc., and may be manufactured into a composite formulation together with other active agents to have a synergistic effect of the active ingredients.

[0081] In addition, according to one specific example of the present invention, the present invention provides the use of a compound represented by the above chemical formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, for use in the manufacture of a drug for the prevention or treatment of cancer. The type of cancer is as mentioned above.

[0082] In addition, according to one specific example of the present invention, the present invention provides a method for treating or preventing an NLRP3-related disease, comprising administering a therapeutically effective amount of a compound represented by the above chemical formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof to a subject in need thereof. The subject may be a mammal including a human.

[0083] In addition, according to one specific example of the present invention, the present invention provides a method for inhibiting NLRP3, comprising a step of administering a therapeutically effective amount of a compound represented by the above chemical formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof to a subject in need thereof.

[0084] The term "therapeutically effective amount" as used herein refers to the amount of the compound represented by the above chemical formula 1 that is effective in treating or preventing a NLRP3-related disease. Specifically, a "therapeutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dosage level can be determined according to factors including the subject type and severity, age, sex, type of disease, activity of the drug, sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field. The pharmaceutical composition of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with commercially available therapeutic agents. And it can be administered singly or in multiple doses. It is important to administer an amount that can achieve the maximum effect with the minimum amount without side effects by taking all of the above factors into consideration, and can be easily determined by those skilled in the art. The dosage of the pharmaceutical composition of the present invention can be determined by an expert according to various factors such as the patient's condition, age, sex, and complications. Since the effective ingredient of the pharmaceutical composition of the present invention has excellent safety, it can be used in amounts exceeding the determined dosage.

[0085] As used herein, “prevention” means any action that inhibits or delays the occurrence, spread, and recurrence of the disease by administering the compound, and “treatment” means any action that improves or beneficially changes the symptoms of the disease by administering the compound.

[0086] In addition, according to one specific embodiment of the present invention, the pharmaceutical composition may further comprise a pharmaceutically acceptable carrier, diluent, or excipient. In one embodiment, the present invention provides a pharmaceutical composition comprising a compound represented by Chemical Formula 1, or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable additive.

[0087] Examples of additives used in the above pharmaceutical composition may include sweeteners, binders, solvents, solubilizers, wetting agents, emulsifiers, isotonic agents, absorbents, disintegrants, antioxidants, preservatives, lubricants, fillers, flavoring agents, and the like. For example, the additives may include lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, silica, talc, stearic acid, stearin, magnesium stearate, magnesium aluminosilicate, starch, gelatin, gum tragacanth, alginic acid, sodium alginate, methylcellulose, sodium carboxymethylcellulose, agar, water, ethanol, polyethylene glycol, polyvinylpyrrolidone, sodium chloride, calcium chloride, orange essence, strawberry essence, vanilla flavoring, and the like.

[0088] The above pharmaceutical composition may be formulated in various formulations for oral administration (e.g., tablets, pills, powders, capsules, syrups or emulsions) or parenteral administration (e.g., intramuscular, intravenous or subcutaneous injection).

[0089] For example, the pharmaceutical composition may be formulated as a preparation for oral administration, and the additives used in this case may 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. Specifically, solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations may be formulated by mixing at least one excipient, for example, starch, calcium carbonate, sucrose, lactose, gelatin, etc., into the composition. In addition to simple excipients, lubricants such as magnesium stearate and talc may be used. In addition, liquid preparations for oral administration may include suspensions, emulsions, syrups, etc., and may include various excipients such as wetting agents, sweeteners, fragrances, and preservatives in addition to commonly used simple diluents such as water and liquid paraffin.

[0090] Additionally, preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solutions and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases may include withepsol, macrogol, Tween 61, cacao butter, laurin, and glycerogelatin. Meanwhile, injections may include conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers, and preservatives.

[0091] Additionally, it can be manufactured as a compound preparation with other active ingredients to have a synergistic effect of the active ingredients.

[0092] The matters mentioned in the uses, compositions, and treatment methods of the present invention apply equally unless they are contradictory.

[0093] The novel compound of the present invention, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof exhibits excellent inhibitory activity against NLRP3 and can therefore be usefully used for the treatment or prevention of NLRP3-related diseases, such as autoimmune diseases, cancer or tumors, and inflammatory diseases.

[0094] Hereinafter, the composition and effects of the present invention will be described in more detail through examples and experimental examples. These examples and experimental examples are intended solely to illustrate the present invention and are not intended to limit the scope of the present invention. All references cited throughout this application are expressly incorporated herein by reference in their entirety.

[0095] The compound of the present invention was purified and its structure was analyzed according to the following method.

[0096] analytical instrument

[0097] LCMS: Agilent Single Quad system (1260), SHIMADZU LCMS-2020

[0098] NMR: BRUKER Advance Nanobay 400MHz, BRUKER AVANCE III / 400

[0099] HPLC: Agilent 1260 II LC, Agilent 1200 / G6410B

[0100] Prep HPLC: Agilent 1290 Infinity II Prep LC

[0101] LCMS analysis method

[0102] LCMS data were recorded on an Agilent Single Quad system (1260) or SHIMADZU LCMS-2020 equipped with an electron spray ionization device. 0.0375% TFA in water (solvent A) and 0.01875% TFA in acetonitrile (solvent B) were used as mobile phases. The columns used were Poroshell 120 EC-C18 (2.1*50) mm, 2.7 μm, Kinetex® EVO C18 2.1 x 30 mm 5 μm, or HALO C18 3.0 x 30 mm, 2.7 μm.

[0103] HPLC analysis method

[0104] HPLC was performed on an Agilent 1260 II LC or Agilent 1200 / G6410B, with mobile phases of 0.0375% TFA in water (solvent A) and 0.01875% TFA in acetonitrile (solvent B). The columns used were Zobrax Eclipse Plus C18 (4.6*150) mm, 3.5 μm or YMC ODS A 4.6*150 mm 3 μm.

[0105] NMR analysis method

[0106] 1 H NMR spectra were recorded using a BRUKER Advance Nanobay 400 MHz / 5 mm Probe (BBO) or BRUKER AVANCE ² 400 MHz / 5 mm Probe (BBO).

[0107] Prep-HPLC separation method

[0108] Prep-HPLC was performed using an Agilent 1290 infinity II, with 0.1% FA in water (solvent A) and 0.1% FA in acetonitrile (solvent B) as mobile phases. The column used was an Agilent 10 Prep-C18 150*30 mm, 10 μm.

[0109] Example: Preparation of the compound of the present invention

[0110] Example 1. Preparation of N-((1H-indazol-7-yl)methyl)-2-ethoxy-3'-methyl-N-((tetrahydrofuran-3-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 1)

[0111]

[0112] [Step 1] Synthesis of 1-(tetrahydrofuran-3-yl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)methanamine (2)

[0113] 1-((2-(Trimethylsilyl)ethoxy)methyl)-1H-indazole-7-carbaldehyde (50 mg, 0.181 mmol, 1 eq) was dissolved in DCM (1 mL), and (tetrahydrofuran-3-yl)methanamine (0.055 mL, 0.543 mmol, 3 eq) was added. The reaction solution was cooled to 0°C, and TEA (0.126 mL, 0.904 mmol, 5 eq) was added. The mixture was stirred at room temperature for 3 h, and after the temperature was lowered to 0°C, STAB (115 mg, 0.543 mmol, 3 eq) was added, and the mixture was stirred at room temperature for 1 h. The main peak of the desired mass was detected by LCMS. The residue was filtered through celite and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (MeOH:DCM=1:20 to 1:10) to obtain the title compound (336 mg, 114% yield). MS[M+H] + = 362.2

[0114] [Step 2] Synthesis of 2-ethoxy-3'-methyl-N-((tetrahydrofuran-3-yl)methyl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (4)

[0115] To a solution of 2-ethoxy-3'-methyl-[1,1'-biphenyl]-4-carboxylic acid (48.2 mg, 0.167 mmol, 1.2 eq) in DMF (1.3 mL) were added HATU (106 mg, 0.279 mmol, 2 eq) and TEA (0.058 mL, 0.418 mmol, 3 eq), and the mixture was stirred at room temperature for 10 min. To the reaction solution was added 1-(tetrahydrofuran-3-yl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)methanamine (50.4 mg, 0.139 mmol, 1 eq) obtained in Step 1, and the mixture was stirred at room temperature for 1 h. The main peak of the desired mass was confirmed by LCMS, and the reaction solution was extracted with EA and brine. The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EA:Hex=70:30 to 50:50) to obtain the title compound (40.4 mg, 48.3% yield). MS[M+H] + = 600.3

[0116] [Step 3] Synthesis of N-((1H-indazol-7-yl)methyl)-2-ethoxy-3'-methyl-N-((tetrahydrofuran-3-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 1)

[0117] 2-Ethoxy-3'-methyl-N-((tetrahydrofuran-3-yl)methyl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (40.4 mg, 0.0674 mmol, 1 eq) obtained in step 2 was dissolved in ACN (0.5 mL), and the temperature of the solution was cooled to 0°C, and BF3·OEt2 (84.6 μL, 0.674 mmol, 10 eq) was added. The mixture was stirred at room temperature for 1 h. The main peak of the desired mass was confirmed by LCMS, and the residue was purified by prep-HPLC to obtain the title compound (12.1 mg, 36.9% yield, 96.5% purity). MS[M+H]+ = 470.2

[0118] 1 H NMR (400 MHz, CDCl3) δ = 11.80 (s, 1H), 8.14 (s, 1H), 7.80 (d,J= 8.0 Hz, 1H), 7.41-7.27 (m, 5H), 7.20-7.11 (m, 2H), 7.01-6.94 (m, 2H), 5.09 (s, 2H), 4.10-3.99 (m, 1H), 3.69-3.62 (m, 4H), 3.37 (d,J= 7.6 Hz, 4H), 2.65-2.59 (m, 1H), 2.42 (s, 3H), 1.83 (m, 1H), 1.36 (t,J= 7.0 Hz, 3H).

[0119] Example 2. Preparation of N-((1H-indazol-7-yl)methyl)-2-ethoxy-3'-methyl-N-((tetrahydrofuran-2-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 2)

[0120]

[0121] [Step 1] Synthesis of 1-(tetrahydrofuran-2-yl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)methanamine (2)

[0122] The title compound (65.6 mg, crude) was obtained by synthesis in a similar manner to step 1 of Example 1. MS[M+H] + = 362.2

[0123] [Step 2] Synthesis of 2-ethoxy-3'-methyl-N-((tetrahydrofuran-2-yl)methyl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (4)

[0124] The title compound (36.7 mg, 33.7% yield) was obtained by synthesis in a similar manner to step 2 of Example 1. MS[M+H] + = 600.4

[0125] [Step 3] Synthesis of N-((1H-indazol-7-yl)methyl)-2-ethoxy-3'-methyl-N-((tetrahydrofuran-2-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 2)

[0126] The title compound (7.8 mg, 27.1% yield, 99.7% purity) was obtained by a similar method to step 3 of Example 1. MS[M+H] + = 470.2

[0127] 1 H NMR (400 MHz, CDCl3) δ = 11.91 (s, 1H), 8.13 (s, 1H), 7.77 (d,J= 8.1 Hz, 1H), 7.40-7.29 (m, 5H), 7.20-7.10 (m, 2H), 7.07-7.01 (m, 2H), 5.24 (s, 2H), 4.29-4.25 (m, 1H), 4.03 (q,J= 7.4 Hz, 2H), 3.92-3.88 (m, 2H), 3.34-3.29 (m, 2H), 2.41 (s, 3H), 1.93-1.88 (m, 4H), 1.37 (t,J= 7.0 Hz, 3H).

[0128] Example 3. Preparation of N-((1H-indazol-7-yl)methyl)-2-ethoxy-3'-methyl-N-((tetrahydro-2H-pyran-4-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 3)

[0129]

[0130] [Step 1] Synthesis of 1-(tetrahydro-2H-pyran-4-yl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)methanamine (2)

[0131] The title compound (63.9 mg, crude) was obtained by synthesis in a similar manner to step 1 of Example 1. MS[M+H] + = 376.2

[0132] [Step 2] Synthesis of 2-ethoxy-3'-methyl-N-((tetrahydro-2H-pyran-4-yl)methyl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (4)

[0133] The title compound (36.4 mg, 34.9% yield) was obtained by synthesis in a similar manner to step 2 of Example 1. MS[M+H] + = 614.5

[0134] [Step 3] Synthesis of N-((1H-indazol-7-yl)methyl)-2-ethoxy-3'-methyl-N-((tetrahydro-2H-pyran-4-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 3)

[0135] The title compound (16.2 mg, 56.4% yield, 99.85% purity) was obtained by a similar method as step 3 of Example 1. MS[M+H] + = 484.2

[0136] 1 H NMR (400 MHz, CDCl3) δ = 11.80 (s, 1H), 8.14 (s, 1H), 7.80 (d,J= 8.1 Hz, 1H), 7.39-7.27 (m, 5H), 7.21-7.11 (m, 2H), 6.96-6.90 (m, 2H), 5.08 (s, 2H), 4.02 (q,J= 6.9 Hz, 2H), 3.86 (d,J= 11.7 Hz, 2H), 3.28 (d,J= 7.5 Hz, 2H), 3.19 (t,J= 11.6 Hz, 2H), 2.42 (s, 3H), 1.89 (s, 1H), 1.36 (t,J= 6.9 Hz, 3H), 1.28 (s, 2H), 1.13-0.99 (m, 2H).

[0137] Example 4. Preparation of N-((1H-indazol-7-yl)methyl)-N-(azetidin-3-yl)-2-ethoxy-3'-methyl-[1,1'-biphenyl]-4-carboxamide (Compound 4)

[0138]

[0139] [Step 1] Synthesis of tert-butyl 3-(((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)amino)azetidine-1-carboxylate (2)

[0140] The title compound (161 mg, crude) was obtained by synthesis in a similar manner to step 1 of Example 1. MS[M+H] + = 433.2

[0141] [Step 2] Synthesis of tert-butyl 3-(2-ethoxy-3'-methyl-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamide)azetidine-1-carboxylate (4)

[0142] The title compound (191 mg, 76.6% yield) was obtained by synthesis in a similar manner to step 2 of Example 1. MS[M+H] + = 671.1

[0143] [Step 3] Synthesis of N-((1H-indazol-7-yl)methyl)-N-(azetidin-3-yl)-2-ethoxy-3'-methyl-[1,1'-biphenyl]-4-carboxamide (Compound 4)

[0144] The title compound (37.1 mg, 29.6% yield, 99.5% purity) was obtained by a similar method as step 3 of Example 1. MS[M+H] + = 441.2

[0145] 1 H NMR (400 MHz, CDCl3) δ = 8.58 (s, 1H), 8.11 (d,J= 8.1 Hz, 1H), 7.75 (t,J= 8.9 Hz, 1H), 7.34 (d,J= 8.8 Hz, 5H), 7.20-7.09 (m, 3H), 5.32 (s, 2H), 4.99-4.91 (m, 1H), 4.11-4.02 (m, 4H), 4.01-3.95 (m, 2H), 2.41 (s, 3H), 1.40-1.25 (m, 3H).

[0146] Example 5. Preparation of N-((1H-indazol-7-yl)methyl)-N-(azetidin-3-ylmethyl)-2-ethoxy-3'-methyl-[1,1'-biphenyl]-4-carboxamide (Compound 5)

[0147]

[0148] [Step 1] Synthesis of tert-butyl 3-((((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)amino)methyl)azetidine-1-carboxylate (2)

[0149] The title compound (178 mg, crude) was obtained by synthesis in a similar manner to step 1 of Example 1. MS[M+H] + = 447.2

[0150] [Step 2] Synthesis of tert-butyl 3-((2-ethoxy-3'-methyl-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamido)methyl)azetidine-1-carboxylate (4)

[0151] The title compound (188 mg, 69.0% yield) was obtained by synthesis in a similar manner to step 2 of Example 1. MS[M+H] + = 685.2

[0152] [Step 3] Synthesis of N-((1H-indazol-7-yl)methyl)-N-(azetidin-3-ylmethyl)-2-ethoxy-3'-methyl-[1,1'-biphenyl]-4-carboxamide (Compound 5)

[0153] The title compound (70.8 mg, 56.6% yield, 96.9% purity) was obtained by a similar method to step 3 of Example 1. MS[M+H] + = 455.2

[0154] 1H NMR (400 MHz, CDCl3) δ = 8.54 (s, 1H), 8.15-7.94 (m, 1H), 7.71-7.67 (m, 1H), 7.40-7.19 (m, 4H), 7.16-6.93 (m, 2H), 7.02 (s, 2H), 5.01 (s, 2H), 4.15-3.75 (m, 4H), 3.69 (s, 3H), 3.47 (s, 2H), 2.34 (s, 3H), 1.41-1.11 (m, 3H).

[0155] Example 6. Preparation of N-((1H-indazol-7-yl)methyl)-2-ethoxy-3'-methyl-N-(pyrrolidin-3-ylmethyl)-[1,1'-biphenyl]-4-carboxamide (Compound 6)

[0156]

[0157] [Step 1] Synthesis of tert-butyl 3-((((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)amino)methyl)pyrrolidine-1-carboxylate (2)

[0158] The title compound (128 mg, crude) was obtained by synthesis in a similar manner to step 1 of Example 1. MS[M+H] + = 461.2

[0159] [Step 2] Synthesis of tert-butyl 3-((2-ethoxy-3'-methyl-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamido)methyl)pyrrolidine-1-carboxylate (4)

[0160] The title compound (199 mg, 102% yield) was obtained by synthesis in a similar manner to step 2 of Example 1. MS[M+H] + = 699.2

[0161] [Step 3] Synthesis of N-((1H-indazol-7-yl)methyl)-2-ethoxy-3'-methyl-N-(pyrrolidin-3-ylmethyl)-[1,1'-biphenyl]-4-carboxamide (Compound 6)

[0162] The title compound (95.5 mg, 65.1% yield, 90.9% purity) was obtained by a similar method as step 3 of Example 1. MS[M+H] + = 469.2

[0163] 1 H NMR (400 MHz, CDCl3) δ = 8.51 (s, 1H), 8.12-8.08 (m, 1H), 7.78-7.73 (m, 1H), 7.40-7.29 (m, 4H), 7.18-6.90 (m, 4H), 5.10 (q,J= 15.3 Hz, 2H), 4.00 (s, 2H), 3.56-3.52 (m, 1H), 3.46-3.36 (m, 1H), 3.31-2.95 (m, 5H), 2.77-2.47 (m, 1H), 2.39 (s, 3H), 1.93-1.69 (m, 1H), 1.43-1.19 (m, 3H).

[0164] Example 7. Preparation of N-((1H-indazol-7-yl)methyl)-2-ethoxy-3'-methyl-N-((1-methylpyrrolidin-3-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 7)

[0165]

[0166] N-((1H-indazol-7-yl)methyl)-2-ethoxy-3'-methyl-N-(pyrrolidin-3-ylmethyl)-[1,1'-biphenyl]-4-carboxamide (11.1 mg, 0.0237 mmol, 1 eq) was dissolved in DCM (0.5 mL), HCHO (5.3 μL, 0.0711 mmol, 3 eq) was added, and the reaction solution was cooled to 0 °C, followed by the addition of TEA (0.0165 mL, 0.118 mmol, 5 eq). The mixture was stirred at room temperature for 30 min, and after the reaction solution was cooled to 0 °C, STAB (15.1 mg, 0.0711 mmol, 3 eq) was added and stirred at room temperature for 12 h. The main peak of the desired mass was confirmed by LCMS, filtered through Celite, and concentrated under reduced pressure. The residue was purified by prep-HPLC to obtain the title compound (5.4 mg, 46.2% yield, 97.7% purity). MS[M+H] + = 483.2

[0167] 1 H NMR (400 MHz, CDCl3) δ = 8.13 (s, 1H), 7.81-7.74 (m, 1H), 7.45-7.26 (m, 5H), 7.21-7.11 (m, 2H), 7.02-6.97 (m, 2H), 5.11 (s, 2H), 4.04 (q,J= 6.9 Hz, 2H), 3.42 (s, 2H), 2.66 (s, 3H), 2.47-1.98 (m, 8H), 1.95-1.76 (m, 1H), 1.50-1.03 (m, 4H).

[0168] Example 8. Preparation of N-((1H-indazol-7-yl)methyl)-2-ethoxy-N-((1-isopropylpyrrolidin-3-yl)methyl)-3'-methyl-[1,1'-biphenyl]-4-carboxamide (Compound 8)

[0169]

[0170] [Step 1] Synthesis of 1-(1-isopropylpyrrolidin-3-yl)-N-((1-((2-trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)methanamine (2)

[0171] The title compound (96.6 mg, 66.3% yield) was obtained by synthesis in a similar manner to step 1 of Example 1. MS[M+H] + = 403.25

[0172] [Step 2] Synthesis of 2-ethoxy-N-((1-isopropylpyrrolidin-3-yl)methyl)-3'-methyl-N―((1-((2-trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (4)

[0173] The title compound (56.4 mg, 37.5% yield) was obtained by synthesis in a similar manner to step 2 of Example 1. MS[M+H] + = 641.30

[0174] [Step 3] Synthesis of N-((1H-indazol-7-yl)methyl)-2-ethoxy-N-((1-isopropylpyrrolidin-3-yl)methyl)-3'-methyl-[1,1'-biphenyl]-4-carboxamide (Compound 8)

[0175] The title compound (16.3 mg, 34.1% yield, 97.04% purity) was obtained by a similar method as step 3 of Example 1. MS[M+H] + = 511.30

[0176] 1H NMR (400 MHz, MeOD) δ = 8.55 (s, 1H), 8.14 (s, 1H), 7.80 (d,J= 8.1 Hz, 1H), 7.40 (s, 1H), 7.34-7.21 (m, 4H), 7.14 (d,J= 7.8 Hz, 2H), 7.00 (s, 1H), 5.00 (s, 2H), 3.95-3.82 (m, 1H), 3.73 (s, 2H), 3.62-3.53 (m, 2H), 3.24-3.11 (m, 1H) 3.01-2.77 (m, 2H), 2.36 (s, 4H), 2.29-2.15 (m, 1H), 1.91 (s, 1H), 1.36 (s, 6H), 1.11 (s, 4H).

[0177] Example 9. Preparation of N-((1H-indazol-7-yl)methyl)-2-ethoxy-3'-methyl-N-(piperidin-4-ylmethyl)-[1,1'-biphenyl]-4-carboxamide (Compound 9)

[0178]

[0179] [Step 1] Synthesis of benzyl 4-(((tert-butoxycarbonyl)amino)methyl)piperidine-1-carboxylate (2)

[0180] tert-Butyl (piperidin-4-ylmethyl)carbamate (1.0 g, 4.67 mmol, 1.0 eq) and KOH (785 mg, 14 mmol, 3.0 eq) were placed in a reaction vessel and dissolved in THF (20 mL). CbzCl (0.866 mL, 6.07 mmol, 1.3 eq) was added dropwise, and the mixture was stirred at room temperature for 3 h. After removing the solvent by concentration under reduced pressure, the organic layer was extracted with H2O (60 mL) and EA (20 mL x 3), dried over MgSO4, filtered, and concentrated under reduced pressure to obtain the residual compound. The residue was purified by silica gel column chromatography (EA / Hex=25 / 75) to obtain the title compound (1.13 g, 69.5% yield) as a white solid. MS [M+Na] + = 371.2

[0181] [Step 2] Synthesis of benzyl 4-(aminomethyl)piperidine-1-carboxylate hydrochloride (3)

[0182] Benzyl 4-(((tert-butoxycarbonyl)amino)methyl)piperidine-1-carboxylate (1.10 g, 3.16 mmol, 1.0 eq) obtained in step 1 was dissolved in DCM (3 mL), 4N HCl / dioxane (2.37 mL, 9.47 mmol, 3.0 eq) was added, and the mixture was stirred at room temperature for 16 h. The solvent was removed by concentration under reduced pressure, and ether was added. The resulting solid was filtered to obtain the title compound (780 mg, 86.8% yield) as a white solid. MS[M+H] + = 249.2

[0183] [Step 3] Synthesis of benzyl 4-((((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)amino)methyl)piperidine-1-carboxylate (4)

[0184] Benzyl 4-(aminomethyl)piperidine-1-carboxylate hydrochloride (824 mg, 2.89 mmol, 2.0 eq) obtained in step 2, 1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazole-7-carbaldehyde (400 mg, 1.45 mmol, 1.0 eq), and molecular sieve (400 mg) were placed in a reaction vessel and dissolved in DCM (4 mL). TEA (1.01 mL, 7.24 mmol, 5.0 eq) was added dropwise, and the mixture was stirred at room temperature for 3 h. STAB (920 mg, 4.34 mmol, 3.0 eq) was added, and the mixture was stirred at room temperature for 1 h. The reaction solution was filtered to remove the molecular sieve, and concentrated under reduced pressure to remove the solvent. The title compound (530 mg, 72.0% yield) was obtained as a white solid by purification using silica gel column chromatography (MeOH / MC=10 / 90). MS[M+H] + = 509.3

[0185] [Step 4] Synthesis of benzyl 4-((2-ethoxy-3'-methyl-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamido)methyl)piperidine-1-carboxylate (5)

[0186] 2-Ethoxy-3'-methyl-[1,1'-biphenyl]-4-carboxylic acid (401 mg, 1.56 mmol, 1.50 eq) and HATU (792 mg, 2.08 mmol, 2.0 eq) were dissolved in DMF (3 mL), and DIPEA (0.558 mL, 3.13 mmol, 3.0 eq) was added, and the mixture was stirred at room temperature for 5 min. Benzyl 4-((((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)amino)methyl)piperidine-1-carboxylate (530 mg, 1.04 mmol, 1.0 eq) obtained in step 3 was added to the mixture, and the mixture was stirred at room temperature for 16 h. After confirming the mass of the target compound by LCMS, H2O (70 mL) and EA (20 ml x 2) were added to extract the organic layer, dried over MgSO4, and concentrated under reduced pressure to remove the solvent. The residue was purified by silica gel column chromatography (EA / Hex=25 / 75) to obtain the title compound (330 mg, 42.4% yield) as a yellow solid. MS[M+H] + = 747.2

[0187] [Step 5] Synthesis of 2-ethoxy-3'-methyl-N-(piperidin-4-ylmethyl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (6)

[0188] Benzyl 4-((2-ethoxy-3'-methyl-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamido)methyl)piperidine-1-carboxylate (320 mg, 0.0428 mmol, 1.0 eq) obtained in step 4 and Pd / C (400 mg) were dissolved in MeOH (10 mL) in a reaction vessel. After adding H2, the reaction solution was stirred at room temperature for 2 h while bubbling gas. After removing the Pd reagent using Celite filtration, the mixture was concentrated under reduced pressure to obtain the title compound (230 mg, 87.6% yield) as a yellow solid. MS[M+H] + = 613.2

[0189] [Step 6] Synthesis of N-((1H-indazol-7-yl)methyl)-2-ethoxy-3'-methyl-N-(piperidin-4-ylmethyl)-[1,1'-biphenyl]-4-carboxamide (Compound 9)

[0190] 2-Ethoxy-3'-methyl-N-(piperidin-4-ylmethyl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (10 mg, 0.0140 mmol, 1.0 eq) obtained in step 5 was dissolved in ACN (0.3 mL), and after cooling to 0°C, BF3OEt2 was added. The temperature was then raised to room temperature and stirred for 1 hour. After confirming the mass of the target compound by LCMS, the title compound (5.7 mg, 84.2% yield, 95.31% purity) was obtained as a white solid by preparative HPLC and lyophilization. MS[M+H] + = 483.2

[0191] 1H NMR (400 MHz, DMSO-d6) δ = 13.08 (s, 1H), 8.11 (s, 1H), 7.71 (d,J= 12 Hz, 1H) 7.35-6.96 (m, 9H), 4.93 (d,J= 64 Hz, 2H), 4.10 (s, 1H), 3.66-3.40 (m, 4H), 2.36-2.26 (m, 4H), 2.10 (s, 1H), 1.78-1.75 (m, 2H), 1.56-1.43 (m, 1H), 1.39-1.19 (m, 4H), 1.10-1.00 (s, 2H), 0.93-0.84 (m, 1H).

[0192] Example 10. Preparation of N-((1H-indazol-7-yl)methyl)-2-ethoxy-3'-methyl-N-((1-methylpiperidin-4-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 10)

[0193]

[0194] [Step 1] Synthesis of 2-ethoxy-3'-methyl-N-((1-methylpiperidin-4-yl)methyl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (2)

[0195] 2-Ethoxy-3'-methyl-N-(piperidin-4-ylmethyl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (30 mg, 0.0489 mmol, 1.0 eq), molecular sieve (20 mg), and 37% HCHO aqueous solution (0.007 mL, 0.0734 mmol, 1.5 eq) were placed in a reaction vessel, dissolved in DCM (0.5 mL), and cooled to 0°C. TEA (0.034 mL, 0.245 mmol, 5.0 eq) was added dropwise, and the mixture was stirred at room temperature for 3 h. STAB (0.031 g, 0.147 mmol, 3.0 eq) was added and stirred at room temperature for 1 hour. The reaction mixture was filtered to remove molecular sieves, concentrated under reduced pressure, and purified by silica gel column chromatography (MeOH / MC=15 / 85) to obtain the title compound (28 mg, 91.2% yield) as a white solid. MS[M+H] + = 627.3

[0196] [Step 2] Synthesis of N-((1H-indazol-7-yl)methyl)-2-ethoxy-3'-methyl-N-((1-methylpiperidin-4-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 10)

[0197] 2-Ethoxy-3'-methyl-N-((1-methylpiperidin-4-yl)methyl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (28 mg, 0.0447 mmol, 1.0 eq) obtained in step 1 was dissolved in ACN (1.0 mL). After cooling the reaction solution to 0°C, BF3OEt2 (0.055 mL, 0.447 mmol, 10.0 eq) was added, and the mixture was stirred at room temperature for 2 h. The resulting mixture was purified by prep-HPLC to obtain the title compound (5 mg, 22.5% yield, 97.75% purity) as a white solid. MS[M+H] + = 497.3

[0198] 1H NMR (400 MHz, DMSO-d6) δ = 13.05 (s, 1H), 8.11 (s, 1H), 7.71 (d,J= 12 Hz, 1H) 7.35-6.96 (m, 9H), 4.94 (d,J= 64 Hz, 2H), 4.10 (s, 1H), 3.66-3.40 (m, 4H), 2.36-2.26 (m, 4H), 2.10 (s, 1H), 1.86-1.68 (m, 3H), 1.65-1.41 (m, 2H), 1.38-1.21 (m, 4H), 1.10-1.00 (s, 2H), 0.93-0.84 (m, 1H).

[0199] Example 11. Preparation of 2-ethoxy-3'-fluoro-N-((4-fluoro-1H-indazol-7-yl)methyl)-N-((1-methylpiperidin-4-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 11)

[0200]

[0201] [Step 1] Synthesis of 1-(4-fluoro-1H-indazol-7-yl)-N-((1-methylpiperidin-4-yl)methyl)methanamine (3)

[0202] 4-Fluoro-1H-indazole-7-carbaldehyde (300 mg, 1.83 mmol) and (1-methylpiperidin-4-yl)methanamine (468.69 mg, 3.66 mmol) were dissolved in DCM (5 mL), and HOAc (109.76 mg, 1.83 mmol, 104.63 μL) was added. The mixture was stirred at 15 °C for 1 h, and NaBH(OAc)3 (774.75 mg, 3.66 mmol) was added and stirred for an additional 11 h. LCMS confirmed complete consumption of the starting material and confirmed that the desired mass was detected. The reaction mixture was diluted with saturated aqueous Na2CO3 solution (10 mL) at 0 °C and extracted with DCM (25 mL Х 2). The collected organic layer was washed with brine (15 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel column chromatography (ISCO®; 12 g SepaFlash® silica flash column, 0–80% MeOH / EA gradient @ 80 mL / min) to obtain the title compound (180 mg, 586.21 μmol, 32.07% yield, 90% purity) as a yellow oil. MS[M+H] + = 277.3

[0203] [Step 2] Synthesis of 2-ethoxy-3'-fluoro-N-((4-fluoro-1H-indazol-7-yl)methyl)-N-((1-methylpiperidin-4-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 11)

[0204] To a solution of 2-ethoxy-3'-fluoro-[1,1'-biphenyl]-4-carboxylic acid (50 mg, 192.12 μmol) and 1-(4-fluoro-1H-indazol-7-yl)-N-((1-methylpiperidin-4-yl)methyl)methanamine (53.09 mg, 192.12 μmol) obtained in Step 1 were added EDCl (55.24 mg, 288.17 μmol), HOBt (31.15 mg, 230.54 μmol), and DIPEA (74.49 mg, 576.35 μmol, 100.39 μL), and the mixture was stirred at 15°C for 16 h. Complete consumption of the starting material was confirmed by LCMS, and a single peak of the desired mass (57%) was detected. The reaction mixture was diluted with H2O (10 mL) at 0°C and extracted with EtOAc (20 mL X 2). The combined organic layer was washed with brine (15 mL X 2), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by prep-HPLC (column: Waters XBridge 150*25 mm* 5 μm; mobile phase: [water (NH4HCO3) -ACN]; gradient: 50%-70% B over 10 min), and the eluate was lyophilized to obtain the title compound (9.75 mg, 18.05 μmol, 9.39% yield, 96% purity) as a white solid. MS[M+H] + = 519.3.

[0205] 1H NMR (400 MHz, DMSO-d6) δ = 13.31-13.07 (m, 1H), 8.19 (s, 1H), 7.50-7.20 (m, 5H), 7.17-7.10 (m, 1H), 7.06 (br d,J= 7.8 Hz, 2H), 6.91 (dd,J= 7.9, 10.1 Hz, 1H), 4.91 (br s, 2H), 4.06-3.84 (m, 2H), 3.37 (br d,J= 5.6 Hz, 2H), 2.98-2.77 (m, 2H), 2.32 (br s, 3H), 2.24-1.99 (m, 2H), 1.86-1.72 (m, 1H), 1.71-1.54 (m, 2H), 1.31-1.12 (m, 5H).

[0206] Example 12. Preparation of N-((1H-indazol-7-yl)methyl)-2-ethoxy-N-((1-isopropylpiperidin-4-yl)methyl)-3'-methyl-[1,1'-biphenyl]-4-carboxamide (Compound 12)

[0207]

[0208] 2-Ethoxy-3'-methyl-N-(piperidin-4-ylmethyl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (30 mg, 0.0489 mmol, 1.0 eq) and K2CO3 (0.0169 mg, 0.122 mmol, 2.5 eq) were dissolved in DMF (1 mL), 2-iodopropane (0.011 mL, 0.108 mmol, 2.2 eq) was added to the reaction solution, heated to 80 °C, and stirred for 1 h. The organic layer of the mixture was extracted with H2O (60 mL) and EA (20 mL x 2), dried over MgSO4, filtered, and concentrated under reduced pressure to obtain the residual compound. After dissolving in ACN (1.0 mL), BF3OEt2 (0.03 mL, 0.245 mmol, 5.0 eq) was added and stirred at room temperature for 16 h. After confirming the mass of the target compound by LCMS, it was separated and purified using prep-HPLC and lyophilized to obtain the title compound (5.0 mg, 19.1% yield, 98.1% purity) as a white solid. MS[M+H] + = 525.3

[0209] 1 H NMR (400 MHz, CDCl3) δ = 11.83 (s, 1H), 8.14 (s, 1H), 7.79 (d,J= 8.1 Hz, 1H), 7.38-7.29 (m, 5H), 7.20-7.11 (m, 2H), 6.92 (d,J= 9.4 Hz, 2H), 5.07 (s, 2H), 4.02 (q,J= 6.9 Hz, 2H), 3.31 (d,J= 7.4 Hz, 2H), 2.97 (d,J= 11.2 Hz, 3H), 2.42 (s, 3H), 2.14 (t,J= 11.5 Hz, 2H), 1.54 (d,J= 13.4 Hz, 2H), 1.37-1.28 (m, 6H), 1.11-1.09 (m, 6H)

[0210] Example 13. Preparation of N-((1H-indazol-7-yl)methyl)-2-ethoxy-N-((1-ethylpiperidin-4-yl)methyl)-3'-methyl-[1,1'-biphenyl]-4-carboxamide (Compound 13)

[0211]

[0212] The title compound (4 mg, 24% yield, 97.83% purity) was obtained as a white solid by a similar method to step 1 of Example 12. MS[M+H] + = 511.3

[0213] 1 H NMR (400 MHz, CDCl3) δ = 11.81 (s, 1H), 8.13 (s, 1H), 7.79 (d,J= 8.1 Hz, 1H), 7.37-7.27 (m, 5H), 7.20-7.13 (m, 2H), 6.91 (d,J= 5.9 Hz, 2H), 5.07 (s, 2H), 4.01 (q,J= 7.0 Hz, 2H), 3.29 (d,J= 7.5 Hz, 2H), 2.92 (d,J= 11.0 Hz, 2H), 2.42 (s, 3H), 1.85-1.6(m, 5H), 1.49 (d,J=13.1 Hz, 2H), 1.35 (t,J= 6.9 Hz, 3H), 1.28 (s, 1H), 1.12-1.06 (m, 4H).

[0214] Example 14.2 Preparation of ethoxy-N-((4-fluoro-1H-indazol-7-yl)methyl)-N-((1-isopropylpiperidin-4-yl)methyl)-3'-methyl-[1,1'-biphenyl]-4-carboxamide (Compound 14)

[0215]

[0216] [Step 1] Synthesis of 1-(4-fluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)-N-((1-isopropylpiperidin-4-yl)methyl)methanamine (2)

[0217] The title compound (145.0 mg, 30.7% yield) was obtained by synthesis in a similar manner to step 1 of Example 1. MS[M+H] + = 435.20

[0218] [Step 2] Synthesis of 2-ethoxy-N-((4-fluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-N-((1-isopropylpiperidin-4-yl)methyl)-3'-methyl-[1,1'-biphenyl]-4-carboxamide (4)

[0219] The title compound (42.2 mg, 72.7% yield) was obtained by synthesis in a similar manner to step 2 of Example 1. MS[M+H] + = 673.25

[0220] [Step 3] Synthesis of 2-ethoxy-N-((4-fluoro-1H-indazol-7-yl)methyl)-N-((1-isopropylpiperidin-4-yl)methyl)-3'-methyl-[1,1'-biphenyl]-4-carboxamide (Compound 14)

[0221] The title compound (6.50 mg, 17.4% yield, 91.30% purity) was obtained by a similar method to step 3 of Example 1. MS[M+H] + = 543.30

[0222] 1 H NMR (400 MHz, DMSO) δ = 13.41 (s, 1H), 8.24 (s, 1H), 7.42 - 6.92 (m, 9H), 4.89 (d,J= 61.5 Hz, 2H), 4.09 (s, 1H), 2.75 - 2.59 (m, 6H), 2.40 - 2.26 (m, 4H), 2.14 (s, 1H), 1.94 (s, 1H), 1.69 (s, 1H), 1.45 (s, 1H), 1.26 (d,J= 13.6 Hz, 3H), 1.08 (s, 2H), 0.93 (d,J= 38.7 Hz, 6H).

[0223] Example 15. Preparation of 2-ethoxy-3'-fluoro-N-((4-fluoro-1H-indazol-7-yl)methyl)-N-((1-isopropylpiperidin-4-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 15)

[0224]

[0225] [Step 1] Synthesis of 2-ethoxy-3'-fluoro-N-((4-fluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-N-((1-isopropylpiperidin-4-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (2)

[0226] The title compound (41.6 mg, 71.2% yield) was obtained by synthesis in a similar manner to step 2 of Example 1. MS[M+H] + = 677.20

[0227] [Step 2] Synthesis of 2-ethoxy-3'-fluoro-N-((4-fluoro-1H-indazol-7-yl)methyl)-N-((1-isopropylpiperidin-4-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 15)

[0228] The title compound (7.60 mg, 20.4% yield, 90.29% purity) was obtained by a similar method as step 3 of Example 1. MS[M+H] + = 547.30

[0229] 1 H NMR (400 MHz, DMSO) δ = 13.42 (s, 1H), 8.22 (s, 1H), 7.50 - 6.93 (m, 9H), 4.88 (d,J= 64.3 Hz, 2H), 3.77 (s, 2H), 2.75 (s, 1H), 2.63 (s, 3H), 2.25 - 2.08 (m, 2H), 2.03 - 1.82 (m, 2H), 1.76 - 1.53 (m, 2H), 1.46 (s, 1H), 1.34 - 1.21 (m, 3H), 1.10 (s, 1H), 0.93 (d,J= 42.1 Hz, 6H).

[0230] Example 16. Preparation of 2-ethoxy-3',5'-difluoro-N-((4-fluoro-1H-indazol-7-yl)methyl)-N-((1-isopropylpiperidin-4-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 16)

[0231]

[0232] [Step 1] Synthesis of 2-ethoxy-3',5'-difluoro-N-((4-fluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-N-((1-isopropylpiperidin-4-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (2)

[0233] The title compound (34.0 mg, 42.8% yield) was obtained by synthesis in a similar manner to step 2 of Example 1. MS[M+H] + = 695.30

[0234] [Step 2] Synthesis of 2-ethoxy-3',5'-difluoro-N-((4-fluoro-1H-indazol-7-yl)methyl)-N-((1-isopropylpiperidin-4-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 16)

[0235] The title compound (6.50 mg, 22.9% yield, 97.41% purity) was obtained by a similar method to step 3 of Example 1. MS[M+H] + = 565.20

[0236] 1 H NMR (400 MHz, DMSO) δ = 13.42 (s, 1H), 8.21 (s,J= 7.2 Hz, 1H), 7.56-6.91 (m, 8H), 4.88 (d,J= 67.8 Hz, 2H), 3.80 (s, 2H), 2.66 (s, 3H), 2.36-2.13 (m, 2H), 2.06-1.80 (m, 2H), 1.67 (d,J= 42.1 Hz, 2H), 1.47 (s, 1H), 1.35-1.22 (m, 3H), 1.12 (s, 2H), 0.95 (d,J= 46.5 Hz, 6H).

[0237] Example 17. Preparation of N-((4-chloro-1H-pyrazolo[4,3-c]pyridin-7-yl)methyl)-2-ethoxy-3'-fluoro-N-((1-isopropylpiperidin-4-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 17)

[0238]

[0239] [Step 1] Synthesis of 1-(4-chloro-1H-pyrazolo[4,3-c]pyridin-7-yl)-N-((1-isopropylpiperidin-4-yl)methyl)methanamine (2)

[0240] 4-Chloro-1H-pyrazolo[4,3-c]pyridine-7-carbaldehyde (130 mg, 0.716 mmol, 1.0 eq) and (1-isopropylpiperidin-4-yl)methanamine dihydrochloride (168 mg, 0.73 mmol, 1.01 eq) were dissolved in DCM, and TEA (0.5 mL, 3.58 mmol, 5.0 eq) was added, and the mixture was stirred at room temperature for 2 h. STAB (760 mg, 3.58 mmol, 5.0 eq) was added to the reaction mixture, and the mixture was stirred at room temperature for an additional 2 h. After confirming the reaction by TLC, the residue was purified by amine column chromatography (MeOH:DCM=1:20) to obtain the title compound (160 mg, 69.4% yield). MS[M+H] + = 322.10

[0241] [Step 2] Synthesis of N-((4-chloro-1H-pyrazolo[4,3-c]pyridin-7-yl)methyl)-2-ethoxy-3'-fluoro-N-((1-isopropylpiperidin-4-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 17)

[0242] 1-(4-chloro-1H-pyrazolo[4,3-c]pyridin-7-yl)-N-((1-isopropylpiperidin-4-yl)methyl)methanamine (148 mg, 0.461 mmol, 1.0 eq) obtained in step 1, 2-ethoxy-3'-fluoro-[1,1'-biphenyl]-4-carboxylic acid (120 mg, 0.461 mmol, 1.0 eq), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) (133 mg, 0.692 mmol, 1.5 eq), 1-hydroxybenzotriazole hydrate (HOBt) (85 mg, 0.553 mmol, 1.2 eq) were dissolved in DMF (6 mL), and then DIPEA (0.247 mL, 1.38 mmol, 3.0 eq) was added and stirred at room temperature for 14 h. The reaction mixture was treated with EA / NaHCO3(aq), and the organic layer was extracted, dried over MgSO4, and concentrated under reduced pressure. The title compound (100 mg, 37.5% yield, 97.5% purity) was obtained as a white solid by preparative HPLC. MS[M+H] + = 564.2

[0243] 1 H NMR (400 MHz, CDCl3) δ = 12.28 (s, 1H), 8.27 (s, 1H), 8.11 (s, 1H), 7.41-7.31 (m, 4H), 7.06 (t,J= 8.4 Hz, 1H), 6.94-6.86 (m, 2H), 5.00 (s, 2H), 4.09-4.01 (m, 2H), 3.35 (d,J= 7.5 Hz, 2H), 3.03-2.91 (m, 3H), 2.16 (t,J= 11.8 Hz, 2H), 1.57 (d,J= 13.1 Hz, 2H), 1.80-1.70(m, 2H) 1.38-1.28 (m, 4H), 1.11 (d,J= 6.6 Hz, 6H).

[0244] Example 18. Preparation of N-((1H-indazol-7-yl)methyl)-3-ethoxy-N-((1-isopropylpiperidin-4-yl)methyl)-4-(2-methylpyridin-4-yl)benzamide (Compound 18)

[0245]

[0246] [Step 1] Synthesis of 1-(1-isopropylpiperidin-4-yl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)methanamine (2)

[0247] 1-((2-(Trimethylsilyl)ethoxy)methyl)-1H-indazole-7-carbaldehyde (500 mg, 1.81 mmol, 1 eq) was dissolved in DCM (30 mL), and (1-isopropylpiperidin-4-yl)methanamine dihydrochloride (1.24 g, 5.43 mmol, 3 eq) was added. The reaction solution was cooled to 0°C, TEA (1.26 mL, 9.04 mmol, 5 eq) was added, and the mixture was stirred at room temperature for 3 h. The reaction solution was cooled to 0°C, STAB (767 mg, 3.62 mmol, 2 eq) was added, and the mixture was stirred at room temperature for 12 h. The main peak of the desired mass was confirmed by LCMS, and the mixture was filtered through Celite and concentrated under reduced pressure to obtain the title compound (2.8 g, crude). MS[M+H] + = 417.2

[0248] [Step 2] Synthesis of 4-bromo-3-ethoxy-N-((1-isopropylpiperidin-4-yl)methyl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)benzamide (3)

[0249] 4-Bromo-3-ethoxybenzoic acid (532 mg, 2.17 mmol, 1.2 eq) was dissolved in DMF (15 mL), and HATU (1.38 g, 3.62 mmol, 2 eq) and TEA (0.757 mL, 5.43 mmol, 3 eq) were added, and the mixture was stirred at room temperature for 10 min. 1-(1-isopropylpiperidin-4-yl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)methanamine (754 mg, 1.81 mmol, 1 eq) obtained in Step 1 was added to the reaction solution, and the mixture was stirred at room temperature for 1 h. The main peak of the desired mass was confirmed by LCMS, and the reaction solution was extracted with EA and brine. The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EA:Hex=70:30 to 50:50) to obtain the title compound (450 mg, 38.6% yield, 99.0% purity). MS[M+H] + = 643.1

[0250] [Step 3] Synthesis of 3-ethoxy-N-((1-isopropylpiperidin-4-yl)methyl)-4-(2-methylpyridin-4-yl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)benzamide (4)

[0251] 4-Bromo-3-ethoxy-N-((1-isopropylpiperidin-4-yl)methyl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)benzamide (50 mg, 0.062 mmol, 1 eq) obtained in step 2 was dissolved in 1,4-dioxane (1 mL), and then 2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (20.4 mg, 0.093 mmol, 1.5 eq) and Cs2CO3 (60.7 mg, 0.186 mmol, 3.0 eq) dissolved in H2O (0.3 mL) were sequentially added, and after removing the gas, Pd(dppf)Cl2·DCM in N2 (5.1 mg, 0.0062 mmol, 0.1 eq) was added. The reaction solution was heated to 100°C and stirred for 12 hours. The main peak of the desired mass was confirmed by LCMS, and the reaction product was filtered through Celite, extracted with EA and brine. The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (MeOH:DCM=1:20 to 1:10) to obtain the title compound (13.0 mg, 15.9% yield, 50.0% purity). MS[M+H] + = 656.2

[0252] [Step 4] Synthesis of N-((1H-indazol-7-yl)methyl)-3-ethoxy-N-((1-isopropylpiperidin-4-yl)methyl)-4-(2-methylpyridin-4-yl)benzamide (Compound 18)

[0253] 3-Ethoxy-N-((1-isopropylpiperidin-4-yl)methyl)-4-(2-methylpyridin-4-yl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)benzamide (13 mg, 50% purity, 0.00991 mmol, 1 eq) obtained in step 3 was dissolved in ACN (1 mL), and after cooling to 0°C, BF3·OEt2 (12.4 μL, 0.0991 mmol, 10 eq) was added. The mixture was stirred at room temperature for 1 h, and the main peak of the desired mass was confirmed by LCMS. The residue was purified by prep-HPLC to give the title compound (0.9 mg, 17.1% yield, 99.14% purity). MS[M+H] + = 526.2

[0254] 1 H NMR (400 MHz, CDCl3) δ = 8.54 (d,J= 5.4 Hz, 1H), 8.45 (s, 1H), 8.14 (s, 1H), 7.80 (d,J= 8.1 Hz, 1H), 7.39-7.32 (m, 2H), 7.31 (t,J= 5.3 Hz, 1H), 7.15 (t,J= 7.6 Hz, 1H), 6.98-6.89 (m, 2H), 5.07 (s, 2H), 4.05 (q,J= 6.9 Hz, 2H), 3.57-3.38 (m, 1H), 3.31 (d,J= 7.3 Hz, 2H), 3.20-3.06 (m, 5H), 2.62 (s, 3H), 2.23 (t,J= 11.8 Hz, 2H), 1.59-1.51 (m, 2H), 1.37 (t,J= 6.8 Hz, 3H), 1.17 (d,J= 6.5 Hz, 6H).

[0255] Example 19. Preparation of 2-cyclopropoxy-3'-fluoro-N-((4-fluoro-1H-indazol-7-yl)methyl)-N-((1-isopropylpiperidin-4-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 19)

[0256]

[0257] [Step 1] Synthesis of 1-(4-fluoro-1H-indazol-7-yl)-N-((1-isopropylpiperidin-4-yl)methyl)methanamine (2)

[0258] 4-Fluoro-1H-indazole-7-carbaldehyde (1 g, 6.09 mmol, 1.2 eq) and (1-isopropylpiperidin-4-yl)methanamine dihydrochloride (1.14 g, 4.97 mmol, 1.0 eq) were dissolved in DCM (20 mL), and TEA (2.55 mL, 18.3 mmol, 3.7 eq) was added. The mixture was stirred at room temperature for 3 h. Then, STAB (5.16 g, 24.4 mmol, 5.0 eq) was added and stirred at room temperature for an additional 3 h. The reaction mixture was filtered through Celite with methanol and concentrated under reduced pressure to obtain the title compound (1.20 g, crude). MS[M+H] + = 305.3

[0259] [Step 2] Synthesis of 2-cyclopropoxy-3'-fluoro-N-((4-fluoro-1H-indazol-7-yl)methyl)-N-((1-isopropylpiperidin-4-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 19)

[0260] 1-(4-Fluoro-1H-indazol-7-yl)-N-((1-isopropylpiperidin-4-yl)methyl)methanamine (2) (15 mg, 0.0493 mmol, 1.0 eq) obtained in step 1, 2-cyclopropoxy-3'-fluoro-[1,1'-biphenyl]-4-carboxylic acid (13.4 mg, 0.049 mmol, 1.0 eq), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) (14.2 mg, 0.0739 mmol, 1.5 eq), 1-hydroxybenzotriazole hydrate (HOBt) (9.1 mg, 0.0591 mmol, 1.2 eq) were dissolved in DMF (1 mL), and then DIPEA (0.0264 mL, 0.148 mmol, 3.0 eq) was added and stirred at room temperature for 19 hours. After syringe filtration, the mixture was separated and purified by prep-HPLC to obtain the title compound (6 mg, 20.8% yield, 95.6% purity) as a white solid. MS[M+H] + = 559.3

[0261] 1 H NMR (400 MHz, CDCl3) δ = 12.0(s, 1H), 8.20 (s, 1H), 7.42-7.28 (m, 3H), 7.23 (dd,J= 13.4, 8.4 Hz, 3H), 7.05 (d,J= 8.6 Hz, 1H), 6.94 (d,J= 7.8 Hz, 1H), 6.82-6.73 (m, 1H), 5.04 (s, 2H), 3.76 (s, 1H), 3.31 (d,J= 7.4 Hz, 2H), 2.99 (d,J= 4.2 Hz, 3H), 2.17 (t,J= 11.5 Hz, 1H), 1.57 (d,J= 13.3 Hz, 2H), 1.33 (dd,J= 27.4, 12.9 Hz, 4H), 1.11 (d,J= 6.6 Hz, 6H), 0.77 (s, 4H).

[0262] Example 20. Preparation of N-((1H-indazol-7-yl)methyl)-3-ethoxy-N-((1-isopropylpiperidin-4-yl)methyl)-4-(2-(trifluoromethyl)pyridin-4-yl)benzamide (Compound 20)

[0263]

[0264] [Step 1] Synthesis of 1-(1-isopropylpiperidin-4-yl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)methanamine (2)

[0265] 1-((2-(Trimethylsilyl)ethoxy)methyl)-1H-indazole-7-carbaldehyde (500 mg, 1.81 mmol, 1.0 eq) was dissolved in DCM (30 mL), cooled to 0°C, and TEA (1.26 mL, 9.04 mmol, 5.0 eq) and (1-isopropylpiperidin-4-yl)methanamine dihydrochloride (1.24 g, 5.41 mmol, 3.0 eq) were slowly added. The mixture was heated to room temperature and stirred for 3 h. After lowering the temperature of the reaction vessel to 0°C, STAB (767 mg, 3.62 mmol, 2.0 eq) was added and stirred at room temperature for 14 h. The reaction was confirmed by TLC (EA / Hx=1 / 2), and the title compound (2.80 g, crude) was obtained by filtration through Celite and concentration under reduced pressure. MS[M+H] + = 417.3

[0266] [Step 2] Synthesis of 4-bromo-3-ethoxy-N-((1-isopropylpiperidin-4-yl)methyl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)benzamide (3)

[0267] 1-(1-Isopropylpiperidin-4-yl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)methanamine (754 mg, 1.81 mmol, 1.0 eq), HATU (1.38 g, 3.62 mmol, 2.0 eq), and TEA (0.757 ml, 5.43 mmol, 3.0 eq) obtained in Step 1 were dissolved in DMF (15 mL) and stirred at room temperature for 10 minutes. Then, 4-bromo-3-ethoxybenzoic acid (532 mg, 2.17 mmol, 1.2 eq) was added to the reaction solution and stirred at room temperature for 1 hour. The reaction was confirmed by TLC, and the organic layer was extracted after treatment with EA / NaCl(aq), dried over MgSO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (MeOH:DCM=1:20 ~ 1:10) to obtain the title compound (450 mg, 38.6% yield). MS[M+H] + = 643.1

[0268] [Step 3] Synthesis of 3-ethoxy-N-((1-isopropylpiperidin-4-yl)methyl)-4-(2-(trifluoromethyl)pyridin-4-yl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)benzamide (4)

[0269] 4-Bromo-3-ethoxy-N-((1-isopropylpiperidin-4-yl)methyl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)benzamide (100 mg, 0.0932 mmol, 1.0 eq) obtained in step 2, 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-(trifluoromethyl)pyridine (38 mg, 0.14 mmol, 1.5 eq), Cs2CO3 (91 mg, 0.28 mmol, 3.0 eq) were dissolved in 1,4-dioxane (1.3 mL) and water (0.5 mL). After degassing with nitrogen, Pd(dppf)Cl2 .DCM (7.6 mg, 0.009 mmol, 0.1 eq) was added, heated to 100°C, and stirred for 11 h. The reaction was confirmed by TLC, treated with EA / NaCl(aq), and the organic layer was extracted, dried over MgSO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (MeOH:DCM=1:20 ~ 1:10) to obtain the title compound (14 mg, 15.9% yield, 50% purity). MS[M+H] + = 710.30

[0270] [Step 4] Synthesis of N-((1H-indazol-7-yl)methyl)-3-ethoxy-N-((1-isopropylpiperidin-4-yl)methyl)-4-(2-(trifluoromethyl)pyridin-4-yl)benzamide (Compound 20)

[0271] 3-Ethoxy-N-((1-isopropylpiperidin-4-yl)methyl)-4-(2-(trifluoromethyl)pyridin-4-yl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)benzamide (4) (14 mg, 0.0197 mmol, 1.0 eq) obtained in step 3 was dissolved in ACN (1 mL), and BF3OEt2 (0.0073 mL, 0.0592 mmol, 3.0 eq) was added, and the mixture was stirred at room temperature for 1 h. After syringe filtration, the title compound (3.2 mg, 28.0% yield, 98.28% purity) was obtained by prep-HPLC. MS[M+H] + = 580.2.

[0272] 1H NMR (400 MHz, CDCl3) δ = 8.78 (d,J= 5.3 Hz, 1H), 8.15 (s, 1H), 7.95 (s, 1H), 7.81 (d,J= 8.0 Hz, 1H), 7.68 (d,J= 5.3 Hz, 1H), 7.39 (d,J= 7.9 Hz, 2H), 7.16 (t,J= 7.5 Hz, 1H), 7.00-6.91 (m, 2H), 5.08 (s, 2H), 4.07 (d,J= 7.1 Hz, 2H), 3.28 (d,J= 7.6 Hz, 2H), 3.01-2.94 (m, 2H), 2.13 (t,J= 12.0 Hz, 1H), 1.54 (d,J= 12.9 Hz, 4H), 1.39 (t,J= 7.0 Hz, 3H), 1.28 (s, 3H), 1.11 (d,J= 6.9 Hz, 6H).

[0273] Example 21. Preparation of N-((1H-indazol-7-yl)methyl)-N-((1-cyclopropylpiperidin-4-yl)methyl)-2-ethoxy-3'-methyl-[1,1'-biphenyl]-4-carboxamide (Compound 21)

[0274]

[0275] 2-Ethoxy-3'-methyl-N-(piperidin-4-ylmethyl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (30 mg, 0.0489 mmol, 1.0 eq), K2CO3 (0.065 mg, 0.392 mmol, 8.0 eq), KI (0.065 g, 0.392 mmol, 8.0 eq) were dissolved in DMSO (2 mL), and bromocyclopropane (0.055 mL, 0.685 mmol, 14 eq) was added and stirred at 180 oThe mixture was heated to C and stirred for 16 h. The organic layer was extracted with H2O (60 mL) and EA (20 mL x 2), dried over MgSO4, filtered, and concentrated under reduced pressure to obtain the residual compound. After that, ACN (2.0 mL) was added to dissolve it, and BF3OEt2 (0.181 mL, 1.47 mmol, 30.0 eq) was added, and the mixture was stirred at room temperature for 2 h. The mass of the target compound was confirmed by LCMS, and the title compound (2.0 mg, 7.82% yield, 94.80% purity) was obtained as a white solid by preparative HPLC and lyophilization. MS[M+H] + = 523.3

[0276] 1 H NMR (400 MHz, CDCl3) δ = 11.81 (s, 1H), 8.13 (d,J= 11.6 Hz, 1H), 7.79 (t,J= 10.4 Hz, 1H), 7.40-7.32 (m, 5H), 7.30-7.17 (m, 2H), 6.90 (d,J= 10.8 Hz, 2H), 5.06 (d,J= 11.2 Hz, 2H), 4.06-3.96 (m, 2H), 3.30 (s, 2H), 3.07-2.96 (m, 3H), 2.41 (d,J= 12.0 Hz, 3H), 1.90-1.52 (m, 5H), 1.50-1.47 (m, 3H), 1.37-1.28 (m, 4H), 1.28-1.10 (m, 2H).

[0277] Example 22. Preparation of N-((4-chloro-1H-pyrazolo[4,3-c]pyridin-7-yl)methyl)-2-ethoxy-3'-methyl-N-((tetrahydro-2H-pyran-4-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 22)

[0278]

[0279] [Step 1] Synthesis of 1-(4-chloro-1H-pyrazolo[4,3-c]pyridin-7-yl)-N-((tetrahydro-2H-pyran-4-yl)methyl)methanamine (2)

[0280] 4-Chloro-1H-pyrazolo[4,3-c]pyridine-7-carbaldehyde (1) (80 mg, 0.441 mmol, 1.0 eq) and (tetrahydro-2H-pyran-4-yl)methanamine (0.11 mL, 0.881 mmol, 2.0 eq) were dissolved in DCM (3 mL), and TEA (0.184 mL, 1.32 mmol, 3.0 eq) was added and stirred at room temperature for 3 h. Then, STAB (0.467 g, 2.22 mmol, 5.0 eq) was added and stirred at room temperature for an additional 2 h. The title compound (100 mg, 80% yield) was obtained by purification by amine column chromatography (MeOH:DCM=1:20 ~ 1:10). MS[M+H] + = 281.10

[0281] [Step 2] Synthesis of N-((4-chloro-1H-pyrazolo[4,3-c]pyridin-7-yl)methyl)-2-ethoxy-3'-methyl-N-((tetrahydro-2H-pyran-4-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 22)

[0282] The title compound (20.6 mg, 20.3% yield, 99.64% purity) was obtained by synthesis in a similar manner to step 2 of Example 1. MS[M+H] + = 519.10

[0283] 1 H NMR (400 MHz, DMSO) δ = 13.93 (s, 1H), 8.37 (s, 1H), 8.07 (d,J= 86.3 Hz, 1H), 7.31 (d,J= 35.5 Hz, 4H), 7.19-6.98 (m, 3H), 4.93 (d,J= 44.9 Hz, 2H), 4.09 (s, 1H), 3.90-3.76 (m, 3H), 3.17 (s, 1H), 2.34 (s, 3H), 2.09-1.89 (m, 1H), 1.62 (s, 1H), 1.44 (s, 1H), 1.26 (d,J=14.0 Hz, 4H), 1.18-1.08 (m, 2H), 0.93 (s, 2H).

[0284] Example 23. Preparation of N-((4-chloro-1H-pyrazolo[4,3-c]pyridin-7-yl)methyl)-2-ethoxy-3'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 23)

[0285]

[0286] [Step 1] Synthesis of 1-(4-chloro-1H-pyrazolo[4,3-c]pyridin-7-yl)-N-((tetrahydro-2H-pyran-4-yl)methyl)methanamine (2)

[0287] 4-Chloro-1H-pyrazolo[4,3-c]pyridine-7-carbaldehyde (80 mg, 0.441 mmol, 1.0 eq) and (tetrahydro-2H-pyran-4-yl)methanamine (0.11 mL, 0.881 mmol, 2.0 eq) were dissolved in DCM (3 mL), and TEA (0.184 mL, 1.32 mmol, 3.0 eq) was added. The mixture was stirred at room temperature for 3 h. Then, STAB (0.467 g, 2.22 mmol, 5.0 eq) was added and stirred at room temperature for 2 h. The reaction mixture was purified by amine column chromatography (MeOH:DCM=1:20 ~ 1:10) to obtain the title compound (100 mg, 80% yield). MS[M+H] + = 281.10

[0288] [Step 2] Synthesis of N-((4-chloro-1H-pyrazolo[4,3-c]pyridin-7-yl)methyl)-2-ethoxy-3'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (Compound 23)

[0289] The title compound (13.0 mg, 12.3% yield, 96.61% purity) was obtained by synthesis in a similar manner to step 2 of Example 1. MS[M+H] + = 523.10

[0290] 1H NMR (400 MHz, DMSO) δ = 13.93 (s, 1H), 8.37 (s, 1H), 8.07 (d,J= 85.5 Hz, 1H), 7.36 (d,J= 52.9 Hz, 4H), 7.21-6.98 (m,J= 53.5 Hz, 3H), 4.93 (d,J= 49.1 Hz, 2H), 4.12 (s, 1H), 3.81 (d,J= 42.4 Hz, 3H), 3.16 (s, 1H), 2.21-1.98 (m, 1H), 1.96-1.87 (m, 1H), 1.62 (s, 1H), 1.43 (s, 1H), 1.27 (d,J= 21.1 Hz, 4H), 1.21-1.11 (m, 2H), 0.94 (s, 1H).

[0291] Example 24. Preparation of N-((1H-indazol-7-yl)methyl)-N-((1-(4-aminophenyl)piperidin-4-yl)methyl)-2-ethoxy-3'-methyl-[1,1'-biphenyl]-4-carboxamide (Compound 24)

[0292]

[0293] [Step 1] Synthesis of tert-butyl ((1-(4-nitrophenyl)piperidin-4-yl)methyl)carbamate (2)

[0294] tert-Butyl (piperidin-4-ylmethyl)carbamate (1.5 g, 7.00 mmol, 1.5 eq) was dissolved in DMF (5 mL), 1-fluoro-4-nitro-benzene (0.45 mL, 4.67 mmol, 1.0 eq) and DIPEA (1.67 mL, 9.33 mmol, 2 eq) were added, and the mixture was heated to 100°C and stirred for 1 h. The mass of the desired compound was confirmed by LCMS, and the mixture was cooled to room temperature. The organic layer was extracted with H2O (50 mL) and EA (30 mL x 2). The extracted organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure to obtain the title compound (1.5 g, crude) as a yellow oil. MS[M+H] + = 336.1

[0295] [Step 2] Synthesis of (1-(4-nitrophenyl)piperidin-4-yl)methanamine hydrochloride (3)

[0296] Tert-Butyl ((1-(4-nitrophenyl)piperidin-4-yl)methyl)carbamate (1.5 g, 4.47 mmol, 1.0 eq) obtained in step 1 was dissolved in DCM (10 mL), 4N HCl / dioxane (5.59 mL, 22.4 mmol, 5.0 eq) was added, and the mixture was stirred at room temperature for 16 h and concentrated under reduced pressure. After MeOH (3 mL) was added to dissolve the mixture, EA (70 mL) was added, and the resulting solid was filtered to obtain the title compound (1.53 g, crude) as a white solid. MS[M+H] + = 236.1

[0297] [Step 3] Synthesis of 1-(1-(4-nitrophenyl)piperidin-4-yl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)methanamine (4)

[0298] 1-((2-(Trimethylsilyl)ethoxy)methyl)-1H-indazole-7-carbaldehyde (1.0 g, 3.62 mmol, 1.0 eq) and (1-(4-nitrophenyl)piperidin-4-yl)methanamine hydrochloride (1.18 g, 4.34 mmol, 1.2 eq) obtained in step 2 were placed in a reaction vessel, dissolved in DCM (12 mL), and TEA (2.52 mL, 18.1 mmol, 5.0 eq) was added dropwise, and stirred at room temperature for 3 h. Then, STAB (2.3 g, 10.9 mmol, 3.0 eq) was added, and stirred for an additional 1 h at room temperature. The compound of the desired mass was confirmed by LCMS, the solvent was removed to obtain the residue, and the residue was purified by silica gel column chromatography (MeOH / MC=5 / 95) to obtain the title compound (1.70 g, 94.8% yield). MS[M+H] + = 496.3

[0299] [Step 4] Synthesis of 2-ethoxy-3'-methyl-N-((1-(4-nitrophenyl)piperidin-4-yl)methyl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (5)

[0300] 2-Ethoxy-3'-methyl-[1,1'-biphenyl]-4-carboxylic acid (1.05 g, 4.12 mmol, 1.2 eq) and HATU (1.96 g, 5.14 mmol, 1.5 eq) were dissolved in DMF (10 mL), and DIPEA (1.84 mL, 10.3 mmol, 3.0 eq) was added, and the mixture was stirred at room temperature for 5 min. 1-(1-(4-nitrophenyl)piperidin-4-yl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)methanamine (1.7 g, 3.43 mmol, 1.0 eq) obtained in Step 3 was added to the mixture, and the mixture was stirred at room temperature for 16 h. Afterwards, the organic layer was extracted with H2O (100 mL) and EA (40 mL x 2). The extracted organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure to obtain the residue, which was then purified by silica gel column chromatography (MeOH / MC=10 / 90) to obtain the title compound (700 mg, 27.8% yield) as a yellow solid. MS[M+H] + = 734.3

[0301] [Step 5] Synthesis of N-((1-(4-aminophenyl)piperidin-4-yl)methyl)-2-ethoxy-3'-methyl-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (6)

[0302] 2-Ethoxy-3'-methyl-N-((1-(4-nitrophenyl)piperidin-4-yl)methyl)-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (450 mg, 0.613 mmol, 1.0 eq) obtained in step 4 and Pd / C (1.0 g) were dissolved in MeOH (10 mL) and DCM (6 mL). After adding H2, the reaction solution was stirred at room temperature for 2 h while injecting gas. After removing the Pd reagent using Celite filtration, the mixture was concentrated under reduced pressure to obtain the title compound (413 mg, 95.7% yield) as a dark blue solid. MS[M+H] + = 704.4

[0303] [Step 6] Synthesis of N-((1H-indazol-7-yl)methyl)-N-((1-(4-aminophenyl)piperidin-4-yl)methyl)-2-ethoxy-3'-methyl-[1,1'-biphenyl]-4-carboxamide (Compound 24)

[0304] N-((1-(4-aminophenyl)piperidin-4-yl)methyl)-2-ethoxy-3'-methyl-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (20 mg, 0.0284 mmol, 1.0 eq) obtained in step 5 was dissolved in ACN (2 mL), and the reaction solution was cooled to 0°C, BF3OEt2 (0.035 mL, 0.284 mmol, 10 eq) was added, and the mixture was stirred at room temperature for 2 h. The molecular weight of the desired compound was confirmed by LCMS, and the mixture was separated and purified using prep-HPLC after syringe filtration and lyophilized to obtain the title compound (1.4 mg, 8.6% yield) as a white solid. MS[M+H] + = 574.3

[0305] 1H NMR (400 MHz, CDCl3) δ = 11.83 (br s, 1H), 8.14 (s, 1H), 7.79 (d,J= 8.1 Hz, 1H), 7.40-7.26 (m, 5H), 7.15 (dd,J= 11.7, 7.3 Hz, 2H), 6.94 (d,J= 7.3 Hz, 2H), 6.79 (d,J= 8.2 Hz, 2H), 6.63 (d,J= 8.2 Hz, 2H), 5.10 (s, 2H), 4.02 (q,J= 7.0 Hz, 2H), 3.33 (d,J= 7.9 Hz, 4H), 2.42 (s, 4H), 1.76 (s, 2H), 1.57 (d,J= 12.8 Hz, 2H), 1.35 (t,J= 6.9 Hz, 4H), 1.28-1.19 (m, 3H)

[0306] Example 25. Preparation of N-((1H-indazol-7-yl)methyl)-N-((1-(4-(dimethylamino)phenyl)piperidin-4-yl)methyl)-2-ethoxy-3'-methyl-[1,1'-biphenyl]-4-carboxamide (Compound 25)

[0307]

[0308] N-((1-(4-aminophenyl)piperidin-4-yl)methyl)-2-ethoxy-3'-methyl-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (20 mg, 0.0284 mmol, 1.0 eq) and molecular sieve 0.3 nm (10 mg) were added to a reaction vessel and dissolved in DCM (1 mL). Then, 37% HCHO aqueous solution (2.3 mg, 0.0284 mmol, 1.0 eq) and TEA (0.0198 mL, 0.142 mmol, 5.0 eq) were added dropwise and stirred at room temperature for 3 h. After that, STAB (0.0181 g, 0.0852 mmol, 3.0 eq) was added and stirred at room temperature for an additional 1 h. The mixture was extracted with saturated aqueous NaHCO3 (60 mL) and EA (20 mL x 2) to extract the organic layer, dried over MgSO4, filtered, and concentrated under reduced pressure to obtain the residual compound. After dissolving in ACN (2 mL), BF3OEt2 (0.00351 mL, 0.0284 mmol, 1.0 eq) was added dropwise, and stirred at room temperature for 1 h. The mass of the target compound was confirmed by LCMS, and the title compound (4.0 mg, 23.4% yield) was obtained as a white solid by separation and purification using prep-HPLC and freeze-drying. MS[M+H] + = 602.3

[0309] 1H NMR (400 MHz, DMSO-d6) δ = 13.06 (s, 1H), 8.12 (d,J= 14.0 Hz, 1H), 7.71 (d,J= 7.9 Hz, 1H), 7.42-7.33 (m, 2H), 7.30-7.10 (m, 6H), 7.10-6.54 (m, 5H), 5.05-4.89 (d,J= 64.8 Hz 2H), 4.11 (s, 1H), 3.66-3.60 (m, 4H), 2.76-2.67 (m, 4H), 1.91-1.73 (m, 1H), 1.46-1.29(m, 2H), 1.28-1.25 (m, 11H), 1.05-0.85 (m, 2H).

[0310] Example 26. Preparation of N-((1H-indazol-7-yl)methyl)-N-((1-(4-acetamidophenyl)piperidin-4-yl)methyl)-2-ethoxy-3'-methyl-[1,1'-biphenyl]-4-carboxamide (Compound 26)

[0311]

[0312] N-((1-(4-aminophenyl)piperidin-4-yl)methyl)-2-ethoxy-3'-methyl-N-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-indazol-7-yl)methyl)-[1,1'-biphenyl]-4-carboxamide (20 mg, 0.0284 mmol, 1.0 eq) and acetic acid (0.00162 mL, 0.0284 mmol, 1.0 eq) were dissolved in DCM (1 mL), and T3P (50% w / w in DCM, 0.0846 mL, 0.142 mmol, 5.0 eq) and DIPEA (0.0254 mL, 0.142 mmol, 5.0 eq) were added dropwise, and the mixture was stirred at room temperature for 16 h. The organic layer was extracted with saturated aqueous NaHCO3 solution (60 mL) and EA (20 mL x 3) from the mixture, dried over MgSO4, filtered, and concentrated under reduced pressure. After dissolving the compound in ACN (1 mL), BF3OEt2 (0.0526 ml, 0.426 mmol, 15 eq) was added, and the mixture was stirred at room temperature for 1 h. The mass of the target compound was confirmed by LCMS, and the mixture was separated and purified using prep-HPLC, and lyophilized to obtain the title compound (8.3 mg, 47.4% yield, 0.2FA salt) as a blue solid. MS[M+H] + = 616.3

[0313] 1H NMR (400 MHz, DMSO-d6) δ = 13.06 (s, 1H), 9.66 (s, 1H), 8.12 (d,J= 14.7 Hz, 1H), 7.71 (d,J= 7.9 Hz, 1H), 7.43 - 7.33 (m, 2H), 7.33 - 7.17 (m, 5H), 7.17 - 6.97 (m, 3H), 6.94 - 6.56 (m, 3H), 5.05 (d,J= 65.6 Hz, 2H), 4.10 (s, 1H), 3.66 - 3.60 (m, 4H), 2.33 - 2.30 (m, 3H), 1.98 (s, 3H), 1.79 (d,J= 12.4 Hz, 2H), 1.57 (d,J= 11.2 Hz, 2H), 1.40 (d,J= 12.1 Hz, 2H), 1.26 (m, 2H), 1.05 (s, 3H).

[0314] <Experimental Example 1> Measurement of IL-1β activity using ELISA

[0315] 1. Culturing of THP-1 cell line

[0316] The THP-1 cell line was purchased from the Korea Cell Line Bank. The cultured cells were maintained at passage 25.

[0317] For cell counting, a Hirschmann hematocytometer (Cat. No. 8100204) and 0.4% trypan blue solution (DyneBio, Cat. No. 15140-122; Lot. No. 20240731) were used.

[0318] For cell culture, RPMI1640 (Gibco, Cat. No. A10491-01; Lot. No. 3062369), FBS (Gibco, Cat. No. 16000-044; Lot. No. U3109271P), penicillin / streptomycin (PS) (Gibco, Cat. No. 15140-122; Lot. No. 227766), 175T cell culture flask (SPL, Cat. No. 71175), 96-well culture plate (SPL, Cat. No. 70075), and PBS pH7.4 (Gibco, Cat. No. 10010-023; Lot. No. 2085080) were used.

[0319] 2. Treatment of the compound of the present invention

[0320] 1 × 10 in each well of a 96-well culture plate 5 The cells were seeded and cultured in a total culture medium volume of 0.15 mL.

[0321] Prior to compound treatment, LPSO111 (Sigma, Cat. L4391; Lot No. 0000297995) was treated to a final concentration of 1 μg / ml and incubated at 37°C for 4 hours to perform the priming process.

[0322] The compounds were completely dissolved in DMSO (Sigma, Cat. No. D2438; Lot. No. RNBJ9566) and used in the experiment. The compounds were diluted three-fold starting from a maximum concentration of 1000 nM, and the concentration of DMSO used in each well was uniformly set to 0.3%.

[0323] Finally, 1 hour after compound treatment, activation was performed by incubating with nigericin sodium salt (Sigma-Aldrich, Cat. No. N7143; Lot. No. 0000308066) at a concentration of 10 μg / ml for 1 hour at 37°C.

[0324] After separating the cells and medium using a centrifuge, only the medium was stored and used in later ELISA experiments.

[0325] 3. Enzyme-linked immunosorbent assay (ELISA)

[0326] To perform the enzyme-linked immunosorbent assay, antibodies were first coated onto an immunoplate (SPL, Cat. No. 32296; Lot. No. BA9E20A32296). The reagents required for each step were from the Human IL-1β ELISA Kit (R&D systems, Cat. No. DY201; Lot. No. P408403). Antibodies were diluted to 4 μg / ml, added to each well, and incubated at room temperature for at least 18 hours.

[0327] After washing each well three times with 0.05% Tween20 (Biorad, Cat. No. 1610781; Lot. No. 64466515) and PBS (Biosesang, Cat. No. PR4007-100-00; Lot. No. P07-122O22DU) buffer, 0.2 ml of 1% BSA (GenDEPOT, Cat. No. A0100-010; Lot. No. 06282420) and PBS buffer was added to each well of each immunoplate, and the blocking process was performed by incubating for 1 hour at room temperature. After removing the buffer, the stored medium was diluted 1 / 30, added to each well, and incubated for 2 hours at room temperature.

[0328] After removing the medium, the wells were washed three times with 0.05% Tween20 and PBS buffer, and then the detection antibody was diluted to 75 ng / ml and added to each well and incubated at room temperature for 2 hours. After washing again with 0.05% Tween20 & PBS buffer, Streptavidin-HRP containing the substrate was diluted 1 / 40 and added to each well, and incubated for 20 minutes at room temperature in a light-shielded manner.

[0329] Finally, after washing three times with 0.05% Tween20 and PBS buffer, 0.1 ml of ultra TMB (Thermo, Cat. No. 34028; Lot. No. AA4536041) was added to each well and incubated for 15 minutes.

[0330] For analysis of the results, a microplate reader image (BMG Labtech, CLARIOstar Plus ) was used to measure the luminescence, and the final measurement values ​​were organized into an Excel file and then displayed as a graph using the Prism-GraphPad program (ver. 9) and IC 50 The values ​​were measured. The results are shown in Table 1 below.

[0331] [Table 1]

[0332]

[0333] As shown in Table 1 above, when the example compounds of the present invention were treated on THP-1 cells, it was confirmed that the activity of IL-1β was effectively reduced, thereby confirming that the example compounds of the present invention are effective for NLRP3-related diseases.

[0334] While the present invention has been described in detail through preferred embodiments and experimental examples, the scope of the present invention is not limited to the specific examples and should be interpreted in accordance with the appended claims. Furthermore, those skilled in the art will appreciate that numerous modifications and variations are possible without departing from the scope of the present invention.

Claims

1. A compound represented by the following chemical formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] In the above chemical formula 1, R1 is -H, -C 1-4 Alkyl, -C 1-4 Haloalkyl, or -halo; X1 to X3 are each independently CR X or N; R X is -H, -C 1-4 Alkyl, -C 1-4 Haloalkyl, or -halo; R2 is -H, -C 1-4 Alkyl, -OH, -OC 1-4 Alkyl, -O-(3-6 membered cycloalkyl), or -halo; Z1 to Z5 are each independently CR Z or N; R Z is -H, -C 1-4 Alkyl, -C 1-4 Haloalkyl, -OH, or -halo; Y is -azetidine or -CH2-W, wherein at least one H of the azetidine ring is -C 1-3 may be substituted with alkyl; W is -(4-7 membered heterocycloalkyl), wherein said -(4-7 membered heterocycloalkyl) contains at least one N or O atom, and at least one H of said -(4-7 membered heterocycloalkyl) ring is -C 1-3 alkyl, -(3-6 membered cycloalkyl), -(4-7 membered heterocycloalkyl), -phenyl, or -(5-6 membered heteroaryl), wherein at least one H of the -phenyl or -(5-6 membered heteroaryl) ring is -C 1-3 Alkyl, -O-R1 or -NR1R2; R1 and R2 are each independently -H, -C 1-3 Alkyl, -C(=O)H or -C(=O)-C 1-3 It's alkyl.

2. In paragraph 1, R1 is -H; X1 is CR X and; X2 is CR X or N; X3 is CR X and; R X is -H, -C 1-4 Alkyl, -C 1-4 Haloalkyl, or -haloine; A compound represented by chemical formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

3. In paragraph 1, R2 is -OC 1-4 Alkyl or -O-(3-6 membered cycloalkyl); A compound represented by chemical formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

4. In paragraph 1, Z1, Z2, Z4, or Z5 are each independently CR Z and; Z3 is CR Z or N; R Z is -H, -C 1-4 Alkyl, -C 1-4 Haloalkyl, or -haloine; A compound represented by chemical formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

5. In paragraph 1, Y is -azetidine or -CH2-W, wherein at least one H of the azetidine ring is -C 1-3 may be substituted with alkyl; W is , , , , , or and; R A is -H, -C 1-3 which may be substituted with alkyl, -(3-6 membered cycloalkyl), or -phenyl, wherein one or more H of the -phenyl ring may be substituted with -NR1R2; R1 and R2 are each independently -H, -C 1-3 Alkyl, or -C(=O)-C 1-3 alkyl; A compound represented by chemical formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

6. A compound selected from the group consisting of the following compounds, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof: .

7. A pharmaceutical composition comprising a compound according to any one of claims 1 to 6, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable additive.

8. A pharmaceutical composition for preventing or treating a disease related to NLRP3, comprising a compound according to any one of claims 1 to 6, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

9. In paragraph 8, A pharmaceutical composition wherein the above NLRP3-related disease is at least one selected from the group consisting of central nervous system disease, metabolic disorder, cardiovascular disease, respiratory disease, liver disease, pancreatic disease, kidney disease, intestinal disease, skin disease, musculoskeletal disease, bone disease, eye disease, inflammation after viral infection, autoimmune disease, cancer or tumor, and other inflammatory diseases.

10. Use of a compound according to any one of claims 1 to 6, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, for use in the manufacture of a medicament for use in the treatment or prevention of a disease associated with NLRP3.

11. A method for treating or preventing an NLRP3-related disease, comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of claims 1 to 6, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

12. A method for treating or preventing at least one disease selected from the group consisting of central nervous system diseases, metabolic disorders, cardiovascular diseases, respiratory diseases, liver diseases, pancreatic diseases, kidney diseases, intestinal diseases, skin diseases, musculoskeletal diseases, bone diseases, eye diseases, inflammation after viral infection, autoimmune diseases, cancer or tumors, and other inflammatory diseases, comprising a step of administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of claims 1 to 6, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

Images