Selective inhibitors of ROCK2 protein kinase and their use

Abstract

The present invention relates to novel substituted bicyclic derivatives capable of selectively inhibiting the phosphorylation of myosin light chain phosphate via ROCK2 and / or Rho kinase, compositions comprising the derivatives, methods for preparing the derivatives and / or compositions, and methods for using the derivatives and / or compositions.

Description

References to related applications

[0001] This application claims priority to U.S. Provisional Application No. 63 / 496,815, filed on 18 April 2023, which is incorporated herein by reference in its entirety. [Technical Field]

[0002] The present invention relates to novel substituted bicyclic derivatives capable of selectively inhibiting the phosphorylation of myosin light chain phosphate via ROCK2 and / or Rho kinase, methods for preparing the same, formulation thereof, and use thereof for the preparation of pharmaceuticals, as well as pharmacodynamically acceptable salts, diastereomers, enantiomers, and racemates. [Background technology]

[0003] Rho-related coiled-coil protein kinases (ROCKs / Rho kinases / Rho-associated kinases) are downstream effectors of the small GTPase Rho (Rho A, Rho B, Rho C, and Rho E) and belong to the serine / threonine kinase family. Active GTP-bound Rho mediates several biological functions via ROCKs, including smooth muscle contraction, cell motility, and cell division. ROCK proteins were identified in 1996 as proteins that bind to Rho GTPases. Two proteins were isolated independently as p160 and p164. These were later recognized as ROCK-1 and ROCK-2, respectively, isoforms of Rho-associated kinases. The two isoforms, ROCK-1 (ROCK-b / p160) and ROCK-2 (ROCK-a / p164), share 92% homology in their amino acid sequences. These structures have a catalytic kinase domain located at the N-terminus, followed by a coiled-coil region (600 amino acids), and p164 shares 92% homology in its amino acid sequence. Its structure includes a catalytic kinase domain located at the N-terminus, a coiled-coil region (600 amino acids) containing a Rho-binding domain, and a plextrin homology (PH) domain at the C-terminus. The two exhibit different localizations, and different physiological roles have been identified for each. ROCK-1 transcript (gene on chromosome 18) is universally present and is more prominently expressed in the liver, kidney, spleen, testes, thymus, and blood cells. On the other hand, ROCK-2 mRNA (chromosome 2) is more highly expressed in skeletal muscle and the brain, suggesting that it plays a specific role in these regions.

[0004] ROCK inhibitors have been investigated for use in numerous diseases, including cerebral ischemia, hypertension, erectile dysfunction, glaucoma, osteoporosis, cardiac hypertrophy, diabetic cardiomyopathy, retinopathy, pulmonary hypertension, and arteriosclerosis. However, their practical application is limited due to a lack of understanding regarding the involvement of specific ROCK isoforms. It remains unclear whether isoform-specific targeting or combined ROCK inhibition yields superior therapeutic effects. Some nonspecific ROCK2 inhibitors have shown promising results in certain conditions such as glaucoma and hypertension, although their specificity is incomplete not only for ROCK isoforms but also for other serine / threonine kinases such as PRK2, PKC, cAMP-dependent protein kinases, and citron kinases. However, while multiple study-based studies have resolved many ambiguities regarding the specific functions of ROCK-1 and ROCK-2, further research is needed before isoform-selective ROCK2 inhibitors become clinically useful. Because ROCK plays a central role in actin cytoskeleton formation, complete inhibition of both isoforms is expected to cause adverse events in patients. Therefore, selective ROCK2 inhibition may have a higher specific therapeutic index than dual inhibition of ROCK1 and ROCK2. Thus, understanding the individual functions of each isoform in specific diseases can contribute to improving the safety and specificity of ROCK inhibitors, as well as expanding their therapeutic applications. [Overview of the project] Means for solving the invention

[0005] In one embodiment, a novel chemical compound represented by the following formula (1) is provided, which has the ability to inhibit Rho-related coiled-coil forming protein serine / threonine kinase (ROCK).

[0006] [ka] Here, X, Y, Z 1 , Z 2 , Z 3 , R 1 , R 2 , R 3 and R 4is defined in the detailed description of the invention below.

[0007] In other embodiments, the present invention provides pharmaceutically acceptable salts, diastereomers, enantiomers, racemates, hydrates, solvates, and prodrugs of the novel compounds. The present invention also provides pharmaceutically acceptable salts, hydrates or solvates of diastereomers, enantiomers, or racemates.

[0008] In yet other embodiments, the present invention provides pharmaceutical compositions comprising a compound, a salt, a diastereomer, an enantiomer, a racemate, a hydrate, a solvate, a prodrug, or a combination thereof.

[0009] In yet other embodiments, the present invention provides methods of treating or alleviating a particular ROCK-mediated disease or disorder using a compound, a salt, a diastereomer, an enantiomer, a racemate, a hydrate, a solvate, a prodrug, or a composition. Non-limiting examples of diseases or disorders include cardiovascular, pulmonary, inflammatory, nervous, or proliferative diseases or disorders.

[0010] In yet other embodiments, the present invention provides methods of preparing a compound, a salt, a diastereomer, an enantiomer, a racemate, a hydrate, a solvate, and a prodrug.

BRIEF DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0011] 1. Compound In one embodiment, the present invention provides a compound represented by formula (1), its pharmaceutically acceptable salt, diastereomer, enantiomer, racemate, hydrate, solvate, or prodrug.

[0012]

CHEMICAL FORMULA

[0013] Here, X is CH2 or O, Y is N or CH, Z 1 Z, 2 and Z 3 are each independently N or CH, n is 0, 1, 2 or 3, R 1 is H, C2-C6 alkyl, or CH2C3-C8 cycloalkyl, and C2-C6 alkyl or CH2C3-C8 cycloalkyl may be optionally substituted with one or more suitable substituents such as C1-C3 alkyl, hydroxyl, or C1-C3 alkoxy, R 2 is 5-6 member heteroaryl or bicyclic heteroaryl, and the heteroaryl or bicyclic heteroaryl may be substituted with one or more suitable substituents such as halogen, CN, CHF2, CF3, C1-C3 alkyl, or amino, and the 5-6 member heteroaryl or bicyclic heteroaryl has 1-3 heteroatoms selected from the group consisting of oxygen, nitrogen and combinations thereof, and R 3 and R 4 are each independently H, Cl, F, OH, OCD 3 3, C1-C3 alkyl, or C1-C3 alkoxy, and C1-C3 alkyl or C1-C3 alkoxy may be optionally substituted with one or more suitable substituents such as OH or OMe, OEt, or OPr.

[0014] In some embodiments, R 1 may be H, C2-C6 alkyl, or C3-C8 cycloalkyl, and C2-C6 alkyl and C3-C8 cycloalkyl may be optionally substituted with one or more suitable substituents such as C1-C3 alkyl, hydroxyl, or C1-C3 alkoxy.

[0015] In some embodiments, R 2 may be any of the following groups.

Chemical formula

[0016] In some embodiments, R 3 or R 4 These are independently H, Cl, F, OH, methoxy, and CD. 3 It may be ethoxy or isopropoxy.

[0017] In this disclosure, the articles "a" and "an" are used to indicate that the grammatical object of the article is one or more (i.e., at least one). For example, "an element" means one or more elements.

[0018] In this disclosure, the term “and / or” means either “and” or “or” unless otherwise specified. The use of the term “or” means “and / or” unless it explicitly refers to the options only or the options are mutually exclusive, but this disclosure supports both the definitions of options and “and / or” only.

[0019] "Optional" or "optionally" means that the events or situations described below may or may not occur, and that the descriptions include both cases in which the events or situations occur and cases in which they do not. For example, "optionally substituted aryl" encompasses both "aryl" and "substituted aryl" as defined herein. Those skilled in the art will understand that with respect to any group containing one or more substituents, such groups are not intended to introduce substitutions or substitution patterns that are sterically unrealistic, synthetically impossible, and / or inherently unstable.

[0020] Unless otherwise specified, the term “optionally substituted” means that a group is either unsubstituted or may be substituted with one or more substituents listed for that group (e.g., 0, 1, 2, 3, 4, or 5 or more, or any range derivable therefrom), and that these substituents may be the same or different. In one embodiment, an optionally substituted group has one substituent. In another embodiment, an optionally substituted group has two substituents. In another embodiment, an optionally substituted group has three substituents. In another embodiment, an optionally substituted group has four substituents. In another embodiment, an optionally substituted group has five substituents. For example, an optionally substituted alkyl group may be a fully saturated alkyl chain (i.e., a pure hydrocarbon). Alternatively, the same optionally substituted alkyl group may have substituents other than hydrogen. For example, an alkyl group may be bonded to a halogen atom, a hydroxyl group, or any other substituent listed herein at any position on the chain. Thus, the term “optionally substituted” means that a particular chemical substance may contain other functional groups, but does not necessarily have other functional groups.

[0021] The term "alkyl," used alone or as part of a larger group such as "arylalkyl" or "cycloalkyl," refers to a linear or branched hydrocarbon group having 1 to 10 carbon atoms, or 1 to 8 carbon atoms, or 1 to 6 carbon atoms, or 1 to 4 carbon atoms (unless otherwise specified), including, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, iso-pentyl, n-hexyl, etc. Alkyls may be unsubstituted or substituted with one or more suitable substituents.

[0022] The term "cycloalkyl" refers to monocyclic or polycyclic hydrocarbon ring groups having 3 to 10 or 3 to 7 carbon atoms in the hydrocarbon ring, unless otherwise specified, and includes, for example, cyclopropyl, cycloheptyl, cyclooctyl, cyclodecyl, cyclobutyl, adamantyl, norpinanyl, dekalinyl, norbornyl, cyclohexyl, and cyclopentyl. Cycloalkyl groups may be unsubstituted or substituted with one or more suitable substituents.

[0023] The term "hetero" means that at least one carbon atom in the ring system is substituted with at least one heteroatom, such as nitrogen, sulfur, sulfoxide, sulfone, or oxygen.

[0024] In this specification, the term "halo" includes fluorine, chlorine, bromine, and iodine. In this specification, the term "alkoxy" refers to the alkyl groups described above that are linked via oxygen, examples of which include methoxy, ethoxy, isopropoxy, and tert-butoxy. Furthermore, alkoxy also refers to polyethers such as -O-(CH2)2-O-CH3. The alkoxy may be unsubstituted or substituted with one or more suitable substituents.

[0025] In this specification, the term "aryl" refers to an unsubstituted or substituted aromatic monocyclic or polycyclic group, including, for example, carbocyclic aromatic groups such as phenyl and naphthyl, and heteroaromatic groups such as pyridyl, furanyl, and thiophenyl. The term "aryl" also includes aromatic rings fused to nonaromatic carbocyclic or heterocyclic rings (e.g., phenyl or pyridyl rings). The term "aryl" can be used interchangeably with "aryl ring," "aromatic group," and "aromatic ring." A heteroaryl group has 4 to 14 atoms in a heteroaromatic ring, of which 1 to 9 are independently selected from the group consisting of oxygen, sulfur, and nitrogen. A heteroaryl group has 1 to 3 heteroatoms in an aromatic ring of 5 to 8 members. An aryl group or heteroaryl group can be a monocyclic or bicyclic aromatic group. Typical aryl and heteroaryl groups include, for example, phenyl, quinolinyl, indazoyl, indolyl, dihydrobenzodioxynyl, 3-chlorophenyl, 2,6-dibromophenyl, pyridyl, pyrimidinyl, 3-methylpyridyl, benzothienyl, 2,4,6-tribromophenyl, 4-ethylbenzothienyl, furanyl, 3,4-diethylfuranyl, naphthyl, 4,7-dichloronaphthyl, pyrrole, pyrazole, imidazole, and thiazole groups. The aryl or heteroaryl may be unsubstituted or substituted with one or more suitable substituents.

[0026] In this specification, the terms "hydroxyl" or "hydroxy" refer to -OH. In this specification, the term "amino" refers to -NH2.

[0027] In this specification, the term "hydroxyalkyl" refers to any hydroxyl group derived from an alkyl group. The term "hydroxyalkyl" includes any alkyl group in which one or more hydrogen atoms are substituted with hydroxyl groups. In this specification, the term "arylalkyl" includes any alkyl group in which one or more hydrogen atoms are substituted with an aryl group, such as a benzyl group, a phenethyl group, and so on.

[0028] In this specification, “substituent” refers to a molecular portion covalently bonded to an atom within the molecule in question. For example, a ring substituent may be a halogen, alkyl group, haloalkyl group, or other group covalently bonded to an atom constituting the ring (preferably a carbon or nitrogen atom). Substituents of aromatic groups are generally covalently bonded to the ring carbon atom. The term “substitution” refers to the substitution of a hydrogen atom in the molecular structure with a substituent, so as to obtain a chemically stable compound (i.e., a compound that can be isolated, characterized, and tested for biological activity) that does not exceed the valence of the specified atom.

[0029] As described above, certain groups may be unsubstituted or substituted with one or more suitable substituents (identical or different groups) other than hydrogen at available positions, typically positions 1, 2, 3, 4, or 5. If a particular group is substituted, it is substituted with a substituent at positions 1, 2, 3, or 4, independently selected. Suitable substituents include halogens, alkyls, haloalkyls, aryls, hydroxys, alkoxys, hydroxyalkyls, and aminos.

[0030] In this specification, the term “pharmaceutically acceptable” means a substance, such as a carrier or diluent, that does not inhibit the biological activity or properties of the compounds described herein. Such a substance is administered to an individual without causing undesirable biological effects or harmful interactions with any component of the composition in which it is contained.

[0031] In this specification, the term "pharmaceutically acceptable salt" refers to a formulation of a compound that does not cause significant irritation to the organism to which it is administered and does not inhibit the biological activity and properties of the compounds described herein.

[0032] Pharmaceutically acceptable salt forms include pharmaceutically acceptable acidic / anionic salts or basic / cationic salts (Reference: UK Journal of Pharmaceutical and Biosciences Vol. 2(4), 01-04, 2014, incorporated herein by reference). Pharmaceutically acceptable acidic / anionic salts include acetates, benzenesulfonates, benzoates, bicarbonates, bicarbonates, tartrates, bromides, calcium edetate, camusylates, carbonates, chlorides, citrates, dihydrochlorides, edetates, edisylates, estruates, esylates, fumarates, glyceptates, glucons, glutamates, glycolyl arsanilates, hexylresorcinates, hydrobroms, hydrochlorides, hydroxynaphthoates, These include iodide, isethionate, lactate, lactobionate, malate, maleate, malonate, mandelate, mesylate, methylsulfate, mucinate, napsylate, nitrate, pamoate, pantothenate, phosphate / diphosphate, polygalacturonate, salicylate, stearate, acetate, succinate, sulfate, bisulfate, tannate, tartrate, theoclate, tosylate, and triethiodide salts. Pharmaceutically acceptable basic / cationic salts include sodium, potassium, calcium, magnesium, diethanolamine, N-methyl-D-glucamine, L-lysine, L-arginine, ammonium, ethanolamine, piperazine, and triethanolamine salts.

[0033] Pharmaceutically acceptable salts are formed by the reaction of the free base form of the compound of Formula 1 with a suitable inorganic or organic acid, including but not limited to hydrobromic acid, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, succinic acid, maleic acid, formic acid, acetic acid, propionic acid, fumaric acid, citric acid, tartaric acid, lactic acid, benzoic acid, salicylic acid, glutamic acid, aspartic acid, p-toluenesulfonic acid, benzenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, naphthalenesulfonic acid such as 2-naphthalenesulfonic acid, or hexanoic acid. Acid addition salts of compounds of Formula 1 that are acceptable as pharmaceuticals include, for example, hydrobromide, hydrochloride, sulfate, nitrate, phosphate, succinate, maleate, formate, acetate, propionate, fumarate, citrate, tartrate, lactate, benzoate, salicylate, glutamate, aspartate, p-toluenesulfonate, benzenesulfonate, methanesulfonate, ethanesulfonate, naphthalenesulfonate (e.g., 2-naphthalenesulfonate) or hexanoate.

[0034] The free acid or free base form of the compound of the present invention is obtained by methods known to those skilled in the art (for example, see the Encyclopedia by L.D. Bigley, S.M. Berg, and D.C. Monkhouse for details). See "of Pharmaceutical Technology," edited by J. Swarbrick and JC Boylam, Vol. 13, Marcel Dekker, Inc., 1995, pp. 453-499; the entire contents of the reference are incorporated herein by reference.) These compounds can be prepared from the corresponding base addition salt or acid addition salt forms. For example, the acid addition salt form of the present invention can be converted to the corresponding free base form by treatment with a suitable base (e.g., ammonium hydroxide solution, sodium hydroxide, etc.). The base addition salt form of the present invention can be converted to the corresponding free acid by treatment with a suitable acid (e.g., hydrochloric acid, etc.).

[0035] Prodrug derivatives of the compounds of the present invention can be obtained by methods known to those skilled in the art (for example, for details, see: Saulnier et al. Bioorg. Med. Chem. Letters, 1994, 4, 1985; Daniela See Hartmann Jornada et al. Al., Molecules 2016, 21, 42. The entire contents of the literature are incorporated herein by reference.) Protected derivatives of the compounds of the present invention can be prepared by methods known to those skilled in the art. A detailed description of the techniques applicable to the creation and removal of protecting groups is given in the literature: TW Greene, “Green's Protective Groups in Organic Chemistry,” 4th This information is contained in the edition, John Wiley and Sons, Inc., 2006, and its entirety is incorporated herein by reference.

[0036] The compounds of the present invention can be prepared as individual stereoisomers by reacting their racemic mixture with an optically active separation agent to form a pair of diastereomer compounds, separating the diastereomers, and recovering the optically pure enantiomers. Separation of the enantiomers can be performed using covalent diastereomer derivatives of the compounds of the present invention or dissociable complexes (e.g., crystalline diastereomer salts). Each diastereomer has different physical properties (e.g., melting point, boiling point, solubility, reactivity, etc.), and these differences can be easily utilized for separation. The diastereomers can be separated by chromatography or separation / decomposition techniques based on solubility differences. The optically pure enantiomers are then recovered together with the separation agent by any practical means that does not cause racemization. For a more detailed description of the techniques for separating the stereoisomers of compounds from a racemic mixture, see: Jean Jacques, Andre Collet, and Samuel H. This information is found in "Enantiomers, Racemates and Resolutions" by Wilen (John Wiley And Sons, Inc., 1981), and its entirety is incorporated herein by reference.

[0037] In this specification, the term “solvate” refers to a stoichiometrically variable complex formed by a solute (in this invention, the compound of Formula 1 or a pharmaceutically acceptable salt thereof) and a solvent. For the purposes of this invention, such a solvent must not interfere with the biological activity of the solute. Non-limiting examples of suitable solvents include water, acetone, methanol, ethanol, and acetic acid. Preferably, the solvent used is a pharmaceutically acceptable solvent. Non-limiting examples of suitable pharmaceutically acceptable solvents include water, ethanol, and acetic acid.

[0038] 2. Composition In other embodiments, the present invention provides pharmaceutical compositions comprising compounds, salts, diastereomers, enantiomers, racemates, hydrates, solvates, prodrugs, or pharmaceutically active combinations thereof. The compositions may further contain additional components. Non-limiting examples of additional components include pharmaceutically acceptable carriers, diluents, excipients, and combinations thereof.

[0039] In this specification, “pharmaceutical composition” means a mixture of the compounds described herein with other chemical components such as carriers, stabilizers, diluents, dispersants, suspending agents, thickeners, and / or excipients. In this specification, the term "pharmaceutical combination" means a product obtained by mixing or combining multiple active ingredients.

[0040] In this specification, the term “acceptable” as used with respect to a formulation, composition, or component means that it does not cause a lasting adverse effect on the overall health of the subject being treated.

[0041] In this specification, the term "carrier" refers to a chemical compound or agent that facilitates the uptake of the compounds described herein into cells or tissues. In this specification, the term “diluent” refers to a chemical compound used to dilute the compounds described herein before administration. Diluents may also be used to stabilize the compounds described herein.

[0042] Suitable carriers, diluents, auxiliaries, or excipients that are acceptable as pharmaceuticals for use in the pharmaceutical compositions of the present invention include, for example, tablets (coated tablets), capsules (gelatin), solutions (aqueous solutions or aqueous ethanol solutions), syrups, emulsions, or inhalation powders containing the active substance (various sugars such as lactose and glucose, salts, and mixtures of these excipients), and aerosols (inhalation solutions with or without propellants).

[0043] Suitable additives include, for example, carriers such as water, paraffin (e.g., petroleum fraction), vegetable oil (e.g., peanut oil or sesame oil), monofunctional or polyfunctional alcohols (e.g., ethanol or glycerol), natural mineral powders (e.g., kaolin, clay, talc, chalk), synthetic mineral powders (e.g., highly dispersible silicic acid and silicates), sugars (e.g., sucrose, lactose, and glucose), emulsifiers (e.g., lignin, spent sulfite solution, methylcellulose, starch, and polyvinylpyrrolidone), and pharmaceutically acceptable organic solvents such as lubricants (e.g., magnesium stearate, talc, stearic acid, sodium lauryl sulfate).

[0044] 3. Method of using the compound or composition In other embodiments, the present invention provides methods for treating or alleviating specific protein kinase-mediated diseases or conditions by administering a therapeutically effective amount of a compound, salt, diastereomer, enantiomer, racemate, hydrate, solvate, prodrug, or composition to a subject or patient. In some embodiments, the present invention provides methods for treating or alleviating specific ROCK-mediated diseases or disorders by administering a therapeutically effective amount of a compound, salt, diastereomer, enantiomer, racemate, hydrate, solvate, prodrug, or composition to a subject or patient. In some embodiments, the present invention provides methods for treating or alleviating diseases or disorders in which ROCK is known to be involved.

[0045] In yet another embodiment, the present invention provides a method for inhibiting enzyme activity, particularly ROCK1, ROCK2, PKCδ, PKCθ, PRK1, GSK3, PRK2, NEK1, and NEK4 kinase activity, by administering a therapeutically effective amount of a compound, salt, diastereomer, enantiomer, racemate, hydrate, solvate, prodrug, or composition to a subject or patient.

[0046] In yet another embodiment, the present invention provides a method for inhibiting protein kinase activity (e.g., ROCK kinase activity) in a biological sample by contacting the biological sample with a compound, salt, diastereomer, enantiomer, racemate, hydrate, solvate, prodrug, or composition.

[0047] In this specification, the term "inhibitor" refers to a compound that inhibits one or more kinases described herein. For example, the term "ROCK inhibitor" refers to a compound that inhibits the ROCK receptor or reduces its signaling effect.

[0048] In this specification, the terms “protein kinase-mediated disease” or “disorder, disease, or condition mediated by inappropriate protein kinase activity” refer to any disease condition mediated or regulated by protein kinases as described herein. Such disease conditions include fibrotic diseases; pulmonary fibrosis including cystic pulmonary fibrosis and idiopathic pulmonary fibrosis; radiation-induced lung injury; hepatic fibrosis including cirrhosis; cardiac fibrosis including arterial fibrosis; endocardial fibrosis; former myocardial infarction; arteriosclerosis; atherosclerosis; restenosis; arthritis fibrosis; Crohn's disease; myelofibrosis; Peyronie's disease; nephrogenic systemic fibrosis; progressive extensive fibrosis; retroperitoneal fibrosis; scleroderma / systemic sclerosis; mediastinal fibrosis; keloids and hypertrophy. Coagulant scarring, glial scarring, or renal fibrosis; cardiovascular diseases or disorders such as cerebral vasospasm, hypertension, atherosclerosis, angina pectoris, myocardial infarction, ischemia / reperfusion injury, stroke, and bronchial asthma; glaucoma, premature birth, erectile dysfunction, or renal diseases such as chronic renal failure, chronic nephritis, diabetic nephropathy, and IgA nephropathy; and proliferative disorders including, but not limited to, retinopathy, fibrosis, or invasive / metastatic cancer. These cancers include adenocarcinoma, adrenocortical carcinoma, bladder cancer, bone cancer, brain tumor, breast cancer, oral cancer, cervical cancer, colon cancer, colorectal cancer, endometrial or uterine cancer, squamous cell carcinoma, esophageal cancer, eye cancer, follicular carcinoma, gallbladder cancer, digestive tract cancer, genitourinary tract cancer, glioblastoma, hairy cell carcinoma, head and neck cancer, liver cancer, hepatocellular carcinoma, Hodgkin's disease, keratogenic echinoderm, kidney cancer, large cell carcinoma, colorectal cancer, laryngeal cancer, liver cancer, lung cancer such as lung adenocarcinoma, small cell lung cancer, lung squamous cell carcinoma, non-small cell lung cancer, melanoma, myeloproliferative disorders, neuroblastoma, ovarian cancer, papillary carcinoma, pancreatic cancer, peritoneal cancer, prostate cancer, rectal cancer, salivary gland cancer, sarcoma, squamous cell carcinoma, small cell carcinoma, small intestine cancer, stomach cancer, testicular cancer, thyroid cancer, and vulvar cancer. In certain embodiments, the cancers being treated are melanoma, breast cancer, colon cancer, or pancreatic cancer.

[0049] In this specification, the terms “to treat,” “in treatment,” or “treatment” refer to methods of preventing, reducing, or improving symptoms of a disease or condition, preventing additional symptoms, improving or preventing the underlying metabolic cause of symptoms, suppressing a disease or condition, halting the progression of a disease or condition, alleviating a disease or condition, causing regression of a disease or condition, alleviating a condition caused by a disease or condition, or preventing and / or halting symptoms of a disease or condition.

[0050] In this specification, the terms “subject” or “patient” include mammals and non-mammals. Examples of mammals include, but are not limited to, humans, chimpanzees, apes, cattle, horses, sheep, goats, pigs, rabbits, dogs, cats, rats, mice, and guinea pigs. Examples of non-mammals include, but are not limited to, birds and fish.

[0051] In this specification, the terms “administer” or “to administer” the subject compound refer to providing the compound and / or its prodrug to a subject in need of treatment.

[0052] In this specification, the terms “effective dose” or “therapeutically effective dose” refer to the administration of an amount of the compound disclosed herein that is sufficient to alleviate, to some extent, one or more symptoms of the disease or condition being treated. This results in a reduction and / or alleviation of the signs, symptoms, or causes of the disease, or other desirable changes in the biological system. For example, “effective dose” in therapeutic use refers to the amount of a composition containing the compound disclosed herein that is necessary to produce a clinically significant reduction in the symptoms of the disease. The appropriate “effective” dose in individual cases can be determined using techniques such as dose escalation studies. For example, therapeutically effective doses of the compounds of the present invention are, for example, about 0.01 mg / kg / day to about 1000 mg / kg / day, about 0.1 mg / kg / day to about 500 mg / kg / day, and about 0.1 mg (x2) / kg / day to about 500 mg (x2) / kg / day.

[0053] The compounds of the present invention were screened against a kinase panel and inhibited the activity of at least one kinase on the panel. Examples of kinases include, but are not limited to, ROCK1 and ROCK2.

[0054] The compounds described herein are inhibitors of ROCK kinase activity and have therapeutic effects in the treatment of diseases associated with inappropriate kinase activity, particularly in the treatment and prevention of kinase-mediated disease conditions, including ROCK kinase. Accordingly, the present invention provides methods for modulating, and in particular inhibiting, kinase-mediated signaling cascades. These methods generally involve administering, or contacting, the compounds, salts, diastereomers, enantiomers, racemates, hydrates, solvates, prodrugs, and / or compositions thereof described in the specification to a subject or to cells expressing the kinase, in order to modulate or inhibit the signaling cascade. These methods are also used to modulate, and in particular inhibit, downstream processes or cellular responses induced by the activation of specific kinase signaling cascades. These methods are also carried out in vitro or in vivo as therapeutic approaches toward the treatment or prevention of diseases characterized, caused, or associated with the activation of kinase-dependent signaling cascades.

[0055] In therapeutic applications of the compounds described herein, including the compound of Formula 1, its salts, diastereomers, enantiomers, racemates, hydrates, solvates, or prodrugs, these compounds are administered alone or as part of a pharmaceutical composition in therapeutically effective amounts. Accordingly, this specification provides pharmaceutical compositions comprising at least one of the compounds described herein, including the compound of Formula 1, its pharmaceutically acceptable salts, diastereomers, enantiomers, racemates, hydrates, solvates, or prodrugs, and one or more pharmaceutically acceptable carriers, diluents, auxiliaries, or excipients.

[0056] Furthermore, these compounds and compositions are administered alone or in combination with one or more additional therapeutic agents. Non-limiting examples of additional therapeutic agents include immune checkpoint inhibitors and immunogenic cell death (ICD) inducing chemotherapeutic agents. Non-limiting examples of immune checkpoint inhibitors include PD-1 inhibitors, PD-L1 inhibitors, and CTLA-4 inhibitors. Non-limiting examples of immunogenic cell death (ICD) inducing chemotherapeutic agents include doxorubicin, idarubicin, mitoxantrone, tautomycin, calculin A, salvulinal, oxaliplatin, bleomycin, and cyclophosphamide. Methods of administration of these compounds and compositions include, but are not limited to, intravenous administration, inhalation, oral administration, rectal administration, parenteral administration, intravitreal administration, subcutaneous administration, intramuscular administration, nasal administration, transdermal administration, topical administration, intraocular administration, oral administration, intratracheal administration, bronchial administration, sublingual administration, or ocular administration. The compounds provided herein are administered by known pharmaceutical formulations, including tablets, capsules, elixirs for oral administration, suppositories for rectal administration, sterile solutions or suspensions for parenteral or intramuscular administration, and transdermal formulations such as lotions, gels, ointments, or creams. In some embodiments, such pharmaceutical compositions are formulated in the form of tablets, pills, capsules, liquids, inhalants, nasal sprays, suppositories, solutions, gels, emulsions, ointments, eye drops, or ear drops.

[0057] The therapeutically effective dose varies depending particularly on the indicated disease, the severity of the disease, the age and relative health status of the patient, the potency of the compound administered, the method of administration, and the desired therapeutic effect. The required dose also varies depending on the method of administration, the specific condition being treated, and the desired effect.

[0058] The compound in Formula 1 inhibits one or more protein kinases and is useful in treating protein kinase-mediated diseases and disorders, such as cancer, autoimmune diseases, fibrotic diseases, cardiovascular diseases, and neurodegenerative diseases.

[0059] In this specification, “biological sample” means a sample present outside the body of an animal, and includes, but is not limited to, cell cultures or their extracts, biopsy material or its extracts taken from an animal, blood, saliva, urine, feces, semen, tears, or other bodily fluids or extracts thereof. Inhibition of kinase activity in biological samples, particularly ROCK kinase activity, is useful for a variety of purposes known to those skilled in the art. Examples of such purposes include, but are not limited to, the preservation of biological specimens and biological assays.

[0060] As used herein, the terms “ROCK-mediated disease” or “condition” mean any disease or other adverse condition in which ROCK is known to be involved. ROCK is involved in a variety of important physiological functions in the vascular system, including smooth muscle contraction, cell proliferation such as the proliferation of vascular smooth muscle cells, cell adhesion and migration (Hu & Lee, Expert Opin. Ther. Targets, 9(4): 715-36, 2005; Shimokawa & Takeshita, Arterioscler. Thromb. Vase. Biol. 25(9):1767-75, ROCK is also involved in inflammatory responses caused by leukocyte migration, such as autoimmune diseases and allergic reactions (Wettschureck). (et al., J. Mol. Med. 80:629-38, 2002). Abnormal activation of the Rho / ROCK pathway has been observed in various central nervous system diseases (see Mueller et al., Nature Rev., 4:387-98, 2005). Furthermore, ROCK has been shown to be involved in tumor cell migration and invasion (Riento & Ridley, Nature Rev. 4: 446-56, 2004) and osteoporosis (Ohnaka et al., Biochem. Biophys. Res. Commun. 287(2):337-4, 2001).

[0061] Specifically, the present invention relates to methods for treating or reducing the severity of cardiovascular diseases or disorders such as cerebral vasospasm, hypertension, atherosclerosis, angina pectoris, myocardial infarction, ischemia / reperfusion injury, stroke, bronchial asthma, glaucoma, premature birth, erectile dysfunction, or renal diseases such as chronic renal failure, chronic nephritis, diabetic nephropathy, and IgA nephropathy, as well as proliferative disorders such as retinopathy, fibrosis, or invasive / metastatic cancer. These cancers include adenocarcinoma, adrenocortical carcinoma, bladder cancer, bone cancer, brain cancer, breast cancer, oral cancer, cervical cancer, colon cancer, colorectal cancer, endometrial or uterine cancer, squamous cell carcinoma, esophageal cancer, eye cancer, follicular carcinoma, gallbladder cancer, gastrointestinal cancer, genitourinary cancer, glioblastoma, hairy cell carcinoma, head and neck cancer, liver cancer, hepatocellular carcinoma, Hodgkin's disease, keratosacral cell carcinoma, kidney cancer, large cell carcinoma, colorectal cancer, laryngeal cancer, liver cancer, lung cancer such as lung adenocarcinoma, small cell lung cancer, lung squamous cell carcinoma, non-small cell lung cancer, malignant melanoma, myeloproliferative disorders, neuroblastoma, ovarian cancer, papillary carcinoma, pancreatic cancer, peritoneal cancer, prostate cancer, rectal cancer, salivary gland cancer, sarcoma, squamous cell carcinoma, small cell carcinoma, small intestine cancer, gastric cancer, testicular cancer, thyroid cancer, and vulvar cancer. In certain embodiments, the cancers targeted for treatment are melanoma, breast cancer, colon cancer, or pancreatic cancer.

[0062] In other embodiments, the present invention provides a method for treating a fibrous disease in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound of Formula 1. Non-limiting examples of fibrous diseases include pulmonary fibrosis, including cystic pulmonary fibrosis and idiopathic pulmonary fibrosis; radiation-induced lung injury; hepatic fibrosis, including cirrhosis; cardiac fibrosis, including arterial fibrosis; endocardial myocardial fibrosis; former myocardial infarction; arteriosclerosis; atherosclerosis; restenosis; arthritis fibrosis; Crohn's disease; myelofibrosis; Peyronie's disease; nephrogenic systemic fibrosis; progressive extensive fibrosis; retroperitoneal fibrosis; scleroderma / systemic sclerosis; mediastinal fibrosis; keloids and hypertrophic scars; glial scars; or renal fibrosis.

[0063] In one embodiment, the present invention provides a method for treating a cell proliferation disorder or condition such as cancer, comprising administering to a subject in need of treatment a therapeutically effective amount of a compound of Formula 1, a pharmaceutically acceptable salt thereof, a diastereomer, an enantiomer, a racemate, a hydrate, a solvate, a prodrug, or a pharmaceutical composition or drug thereof, such as lymphoma, osteosarcoma, melanoma, breast cancer, kidney cancer, prostate cancer, colorectal cancer, thyroid cancer, ovarian cancer, pancreatic cancer, nerve cancer, lung cancer, uterine cancer, or gastrointestinal cancer. In one embodiment, the present invention provides a method for inhibiting the proliferation of cancer cells using a compound, a salt, a diastereomer, an enantiomer, a racemate, a hydrate, a solvate, a prodrug, or any combination thereof, or a composition thereof.

[0064] In certain embodiments, the protein kinase-mediated disease or condition is an inflammatory disease or condition, a respiratory disease or autoimmune disease or condition, such as asthma, chronic obstructive pulmonary disease (COPD), adult respiratory distress syndrome (ARDS), ulcerative colitis, Crohn's disease, bronchitis, dermatitis, allergic rhinitis, psoriasis, scleroderma, urticaria, rheumatoid arthritis, multiple sclerosis, cancer, breast cancer, HIV-related disease, or lupus.

[0065] In other embodiments, the present invention provides a method for treating cardiovascular disease by administering to a subject a therapeutically effective amount of a compound, salt thereof, diastereomer, enantiomer, racemate, hydrate, solvate, prodrug, or any combination thereof, or a composition thereof, as described herein. Such cardiovascular diseases affect the heart or blood vessels and include, for example, atherosclerosis, arrhythmia, angina pectoris, myocardial ischemia, myocardial infarction, aneurysm of the heart or blood vessels, vasculitis, stroke, peripheral occlusive arteriovenous disease of the limbs, organs, or tissues, post-ischemia reperfusion injury of organs or tissues, endotoxin shock, surgical shock, or traumatic shock, hypertension, valvular heart disease, heart failure, abnormal blood pressure, vasoconstriction, vascular abnormalities, or inflammation.

[0066] In other aspects, the present invention provides a method for treating cancer, comprising administering to a subject requiring treatment a composition comprising at least one therapeutically effective amount of a compound, salt thereof, diastereomer, enantiomer, racemate, hydrate, solvate, or prodrug described herein and at least one therapeutically effective amount of an immune checkpoint inhibitor, wherein cancer is adenocarcinoma, adrenocortical carcinoma, bladder cancer, bone cancer, brain cancer, breast cancer, oral cancer, cervical cancer, colon cancer, colorectal cancer, endometrial cancer, or uterine cancer. Squamous cell carcinoma, esophageal cancer, eye cancer, follicular carcinoma, gallbladder cancer, gastrointestinal cancer, genitourinary cancer, glioblastoma, hairy cell carcinoma, head and neck cancer, liver cancer, hepatocellular carcinoma, Hodgkin's disease, keratosacral cell carcinoma, kidney cancer, large cell carcinoma, colorectal cancer, laryngeal cancer, liver cancer, lung adenocarcinoma, small cell lung cancer, lung squamous cell carcinoma, non-small cell lung cancer, melanoma, myeloproliferative disorders, neuroblastoma, ovarian cancer, papillary carcinoma, pancreatic cancer, peritoneal cancer, prostate cancer, rectal cancer, salivary gland cancer, sarcoma, squamous cell carcinoma, small cell carcinoma, small intestine cancer, gastric cancer, testicular cancer, thyroid cancer, vulvar cancer, or any combination thereof. Examples of checkpoint inhibitors include, but are not limited to, PD-1 inhibitors, PD-L1 inhibitors, and CTLA-4 inhibitors.

[0067] In other aspects, the present invention provides a method for treating cancer, comprising administering to a subject requiring treatment a composition comprising at least one therapeutically effective amount of a compound, salt thereof, diastereomer, enantiomer, racemate, hydrate, solvate, or prodrug described herein and at least one immunogenic cell death (ICD)-inducing chemotherapeutic agent, wherein cancer is adenocarcinoma, adrenocortical carcinoma, bladder cancer, bone cancer, brain cancer, breast cancer, oral cancer, cervical cancer, colon cancer, colorectal cancer, endometrial cancer, or uterine cancer. Cancer of the cervix, squamous cell carcinoma, esophageal cancer, eye cancer, follicular carcinoma, gallbladder cancer, gastrointestinal cancer, genitourinary cancer, glioblastoma, hairy cell carcinoma, head and neck cancer, liver cancer, hepatocellular carcinoma, Hodgkin's disease, keratospermatoma, kidney cancer, large cell carcinoma, colorectal cancer, laryngeal cancer, liver cancer, lung adenocarcinoma, small cell lung cancer, lung squamous cell carcinoma, non-small cell lung cancer, melanoma, myeloproliferative disorder, neuroblastoma, ovarian cancer, papillary carcinoma, pancreatic cancer, peritoneal cancer, prostate cancer, rectal cancer, salivary gland cancer, sarcoma, squamous cell carcinoma, small cell carcinoma, small intestine cancer, stomach cancer, testicular cancer, thyroid cancer, vulvar cancer, or any combination thereof. Examples of immunogenic cell death (ICD)-inducing chemotherapeutic agents include, but are not limited to, doxycycline, idarubicin, mitoxantrone, tautomycin, calculin A, salvulinal, oxaliplatin, bleomycin, and cyclophosphamide.

[0068] In the above-described method using the compounds of the present invention, the compounds described herein, their salts, diastereomers, enantiomers, racemates, hydrates, solvates, or prodrugs are administered to a system including cells or tissues. In certain embodiments, the compounds described herein, their salts, diastereomers, enantiomers, racemates, hydrates, solvates, prodrugs, or any combination thereof are administered to human or animal subjects. In even more specific embodiments, a pharmaceutical composition or drug comprising at least one of the compounds, salts, diastereomers, enantiomers, racemates, hydrates, solvates, prodrugs, or any combination thereof is administered to human or animal subjects.

[0069] 4. Method for producing the compound In other embodiments, the present invention provides methods for preparing compounds, salts, diastereomers, enantiomers, racemates, hydrates, solvates, and prodrugs. In some embodiments, compounds, salts, diastereomers, enantiomers, racemates, hydrates, solvates, or prodrugs can be prepared by methods including, but not limited to, one or more of the following: (a) If necessary, convert the compound of the present invention into a pharmaceutically acceptable salt. (b) Convert the salt form of the compound of the present invention to the non-salt form as necessary. (c) If necessary, convert the non-oxidized form of the compound of the present invention to a pharmaceutically acceptable N-oxide. (d) If necessary, separate the individual isomers of the compound of the present invention from a mixture of stereoisomers. (e) If necessary, convert the non-derivative compound of the present invention into a pharmaceutically acceptable prodrug derivative, and, (f) If necessary, convert the prodrug derivative of the compound of the present invention to its non-derivative form.

[0070] Exemplary methods for preparing the compounds of the present invention are described herein, including examples that are described in detail below. Some embodiments of the present invention provide methods for preparing the compounds of the present invention, as shown in Method 1 or Method 2 below.

[0071] [ka]

[0072] A bicyclic acid (2) was reacted with thionyl chloride or oxalyl chloride in dichloromethane in the presence of a catalytic amount of GMF to obtain acylchloride (3). The obtained acylchloride (3) was coupled with a bicyclic heteroaryl compound (4) by a Friedel-Crafts acylation reaction in an organic solvent (dichloromethane or dichloroethane) in the presence of a Lewis acid (aluminum chloride, titanium chloride, or tin chloride) to obtain ketone (5). The obtained ketone (5) was reacted with an alkyl bromide, alkyl chloride, or alkyl epoxide in an organic solvent (DMF, acetone, dioxane, acetonitrile, dichloromethane, or dichloroethane) using an inorganic base (NaHCO3, Na2CO3, K2CO3, Cs2CO3, K2HPO4, or K3PO4) at room temperature (RT) to 150°C for 5 to 24 hours to obtain an N-alkylated compound (6).

[0073] The N-alkylated compound (6) could also be prepared by starting with the bicyclic heteroaryl compound (4), performing N-alkylation under the above reaction conditions to obtain (7), and then carrying out a Friedel-Crafts acylation reaction using acylchloride (3).

[0074] The N-alkylated compound (6) was reacted with a boronic acid or boron ester under Suzuki-Hannō conditions using a palladium catalyst (Pd(OAc)2, Pd2(dba)3, Pd(PPh3)4, or Pd(dppf)Cl2·DCM) and reacted at room temperature (RT) to 150°C for 5 to 24 hours in the presence of an inorganic base (NaHCO3, Na2CO3, K2CO3, Cs2CO3, CsF, K2HPO4, or K3PO4) to obtain compound (1).

[0075] The N-alkylated compound (6) was alternatively prepared starting from the bicyclic heteroaryl compound (4) (Method 2).

[0076] [ka]

[0077] The bicyclic heteroaryl compound (4) was reacted with N-iodosuccinimide (NIS) in an organic solvent (DMF, acetone, dioxane, acetonitrile, dichloromethane, or dichloroethane) at room temperature (RT) for 2 to 24 hours to obtain the iodized bicyclic heteroaryl compound (8). The iodized bicyclic heteroaryl compound (8) was reacted with an alkyl bromide, alkyl chloride, or alkyl epoxide, and the reaction was carried out in an organic solvent (DMF, acetone, dioxane, acetonitrile, dichloromethane, or dichloroethane) using an inorganic base (NaHCO3, Na2CO3, K2CO3, Cs2CO3, K2HPO4, or K3PO4) at room temperature (RT) to 150°C for 5 to 24 hours to obtain the N-alkylated compound (10). The N-alkylated compound (10) was alternatively prepared by N-alkylating the bicyclic heteroaryl compound (4) to (9), followed by iodizing (9) with N-iodosuccinimide (NIS) as described above. The bicyclic acid (2) was coupled with N-methoxymethanamine hydrochloride in an organic solvent (DMF, tetrahydrofuran, dichloromethane, or dichloroethane) at room temperature (RT) for 2 to 24 hours using a coupling reagent (DCC, EDCI, HATU, HBTU, PyBop, or PyBrop) to obtain Weinrebamide (11). Weinrebamide (11) was reacted with the iodized bicyclic heteroaryl compound (10) in THF at low temperature (-78 to 0°C) in the presence of a Grignard reagent to convert it to a ketone (6).

[0078] Non-limiting examples of compounds described herein include the following: (6-chlorochroman-3-yl)-[1-(2-hydroxyethyl)-6-(3-methoxy-1H-pyrazole-4-yl)pyrrolo[3,2-c]pyridine-3-yl]methanone; (6-chloro-2H-chromen-3-yl)-[6-(3-chloro-1H-pyrazole-4-yl)-1-(2-hydroxyethyl)pyrrolo[3,2-c]pyridine-3-yl]methanone; (6-(3-chloro-1H-pyrazole-4-yl)-1-(2-hydroxyethyl)-1H-pyrrolo[2,3-b]pyridine-3-yl)(6-fluorochroman-3-yl)methanone; (6-chlorochroman-3-yl)-[6-(3-chloro-1H-pyrazole-4-yl)-1-(2-hydroxyethyl)pyrrolo[2,3-b]pyridine-3-yl]methanone; (6-chlorochroman-3-yl)-[6-(3-chloro-1H-pyrazole-4-yl)-1-(2-hydroxyethyl)pyrrolo[3,2-b]pyridine-3-yl]methanone; (6-chlorochroman-3-yl)-[6-(3-chloro-1H-pyrazole-4-yl)-1-(2-hydroxypropyl)pyrrolo[3,2-c]pyridine-3-yl]methanone; (6-chlorochroman-3-yl)-[6-(5-chloro-1H-pyrazole-4-yl)-1-[(2S)-2-hydroxypropyl]pyrrolo[3,2-c]pyridine-3-yl]methanone; (6-chlorochroman-3-yl)-[6-(3-chloro-1H-pyrazole-4-yl)-1-[(2R)-2-hydroxypropyl]pyrrolo[3,2-c]pyridine-3-yl]methanone; (6-(5-chloro-1H-pyrazole-4-yl)-1-((S)-2-hydroxypropyl)-1H-pyrrolo[3,2-c]pyridine-3-yl)((S)-6-chlorochroman-3-yl)methanone; (6-(5-chloro-1H-pyrazole-4-yl)-1-((S)-2-hydroxypropyl)-1H-pyrrolo[3,2-c]pyridine-3-yl)((R)-6-chlorochroman-3-yl)methanone; (6-(5-chloro-1H-pyrazole-4-yl)-1-((S)-2-hydroxypropyl)-1H-pyrrolo[3,2-c]pyridine-3-yl)((S)-6-chlorochroman-3-yl)methanomethanesulfonic acid; [6-(3-chloro-1H-pyrazole-4-yl)-1-(2-hydroxyethyl)pyrrolo[3,2-c]pyridine-3-yl]-(6-fluorochroman-3-yl)methanone; [6-(3-chloro-1H-pyrazole-4-yl)-1-(2-hydroxypropyl)pyrrolo[3,2-c]pyridine-3-yl]-(6-fluorochroman-3-yl)methanone; (6-Fluorochroman-3-yl)-[1-(2-hydroxyethyl)-6-(3-methyl-1H-pyrazole-4-yl)pyrrolo[2,3-b]pyridine-3-yl]methanone; (6-chlorochroman-3-yl)-[6-(3-chloro-1H-pyrazole-4-yl)-1-(2-hydroxy-2-methyl-propyl)pyrrolo[3,2-c]pyridine-3-yl]methanone; (6-chlorochroman-3-yl)-[6-(3-chloro-1H-pyrazole-4-yl)-1-(2-hydroxy-2-methylpropyl)pyrrolo[3,2-c]pyridine-3-yl]methanone; (6-chlorochroman-3-yl)-[6-(3-chloro-1H-pyrazole-4-yl)-1-(2-hydroxy-2-methylpropyl)pyrrolo[3,2-c]pyridine-3-yl]methanone; (6-chlorochroman-3-yl)-[6-(3-chloro-1H-pyrazole-4-yl)-1-(2-hydroxy-2-methyl-propyl)pyrrolo[3,2-c]pyridine-3-yl]methanone hydrochloride; [6-(3-chloro-1H-pyrazole-4-yl)-1-[(2S)-2-hydroxypropyl]pyrrolo[3,2-c]pyridine-3-yl]-(6-fluorochroman-3-yl)methanone; [[6-(3-chloro-1H-pyrazole-4-yl)-1-[(2R)-2-hydroxypropyl]pyrrolo[3,2-c]pyridine-3-yl]-(6-fluorochroman-3-yl)methanone; [6-(3-chloro-1H-pyrazole-4-yl)-1-(2-hydroxyethyl)pyrrolo[2,3-b]pyridine-3-yl]-(6-methoxychroman-3-yl)methanone; [6-(3-chloro-1H-pyrazole-4-yl)-1-(2-hydroxyethyl)pyrrolo[3,2-c]pyridine-3-yl]-(6-methoxychroman-3-yl)methanone; [6-(5-fluoro-1H-pyrazole-4-yl)-1-(2-hydroxyethyl)pyrrolo[3,2-c]pyridine-3-yl]-(6-methoxychroman-3-yl)methanone; [6-(5-fluoro-1H-pyrazole-4-yl)-1-[(2S)-2-hydroxypropyl]pyrrolo[3,2-c]pyridine-3-yl]-(6-methoxychroman-3-yl)methanone; [6-(5-fluoro-1H-pyrazole-4-yl)-1-[(2S)-2-hydroxypropyl]pyrrolo[3,2-c]pyridine-3-yl]-(6-methoxychroman-3-yl)methanone; [6-(5-chloro-1H-pyrazole-4-yl)-1-[(2S)-2-hydroxypropyl]pyrrolo[3,2-c]pyridine-3-yl]-(6-methoxychroman-3-yl)methanone; [6-(5-fluoro-1H-pyrazole-4-yl)-1-(2-hydroxy-2-methyl-propyl)pyrrolo[3,2-c]pyridine-3-yl]-(6-methoxychroman-3-yl)methanone; [6-(5-chloro-1H-pyrazole-4-yl)-1-(2-hydroxy-2-methyl-propyl)pyrrolo[3,2-c]pyridine-3-yl]-(6-methoxychroman-3-yl)methanone; [(3S)-6-chlorochroman-3-yl]-[6-(5-fluoro-1H-pyrazole-4-yl)-1-[(2S)-2-hydroxypropyl]pyrrolo[3,2-c]pyridine-3-yl]methanone; [(3S)-6-chlorochroman-3-yl]-[6-(5-fluoro-1H-pyrazole-4-yl)-1-(2-hydroxy-2-methylpropyl)pyrrolo[3,2-c]pyridine-3-yl]methanone; [(3S)-6-chlorochroman-3-yl]-[6-(5-chloro-1H-pyrazole-4-yl)-1-(2-hydroxy-2-methylpropyl)pyrrolo[3,2-c]pyridine-3-yl]methanone; [(3S)-6-chlorochroman-3-yl]-[1-[(2S)-2-hydroxypropyl]-6-(5-methoxy-1H-pyrazole-4-yl)pyrrolo[3,2-c]pyridine-3-yl]methanone; [(3R)-6-chlorochroman-3-yl]-[1-[(2R)-2-hydroxypropyl]-6-(5-methoxy-1H-pyrazole-4-yl)pyrrolo[3,2-c]pyridine-3-yl]methanone; (6-chlorochroman-3-yl)-[6-(5-chloro-1H-pyrazole-4-yl)-1-[(2S)-2-hydroxybutyl]pyrrolo[3,2-c]pyridine-3-yl]methanone; (6-chlorochroman-3-yl)-[6-(5-fluoro-1H-pyrazole-4-yl)-1-[(2S)-2-hydroxybutyl]pyrrolo[3,2-c]pyridine-3-yl]methanone; (6-chlorochroman-3-yl)-[6-(5-fluoro-1H-pyrazole-4-yl)-1-[(3-hydroxycyclobutyl)methyl]pyrrolo[3,2-c]pyridine-3-yl]methanone; (6-chlorochroman-3-yl)-[6-(5-chloro-1H-pyrazole-4-yl)-1-[(3-hydroxycyclobutyl)methyl]pyrrolo[3,2-c]pyridine-3-yl]methanone; (6-chlorochroman-3-yl)-[6-(3-chloro-1H-pyrazole-4-yl)-1-(2-hydroxypropyl)indole-3-yl]methanone; (6-(3-chloro-1H-pyrazole-4-yl)-1-(2-hydroxypropyl)-1H-indole-3-yl)((S)-6-chlorochroman-3-yl)methanone; (6-(3-chloro-1H-pyrazole-4-yl)-1-(2-hydroxypropyl)-1H-indole-3-yl)((S)-6-chlorochroman-3-yl)methanone; ((S)-6-chlorochroman-3-yl)(1-(2-hydroxypropyl)-6-(3-methoxy-1H-pyrazole-4-yl)-1H-indole-3-yl)methanone; (6-chlorochroman-3-yl)-[1-[(2S)-2-hydroxypropyl]-6-(3-methoxy-1H-pyrazole-4-yl)indole-3-yl]methanone; (6-chlorochroman-3-yl)-[1-[(2R)-2-hydroxypropyl]-6-(3-methoxy-1H-pyrazole-4-yl)indole-3-yl]methanone; (6-chlorochroman-3-yl)-[2-(5-chloro-1H-pyrazole-4-yl)-7-[(2S)-2-hydroxypropyl]pyrrolo[2,3-d]pyrimidine-5-yl]methanone; (2-(5-chloro-1H-pyrazole-4-yl)-7-((S)-2-hydroxypropyl)-7H-pyrrolo[2,3-d]pyrimidine-5-yl)((S)-6-chlorochroman-3-yl)methanone; (6-chlorochroman-3-yl)-[2-(5-fluoro-1H-pyrazole-4-yl)-7-[(2S)-2-hydroxypropyl]pyrrolo[2,3-d]pyrimidine-5-yl]methanone; (2-(5-fluoro-1H-pyrazole-4-yl)-7-((R)-2-hydroxypropyl)-7H-pyrrolo[2,3-d]pyrimidine-5-yl)(6-fluorochroman-3-yl)methanone; (2-(5-fluoro-1H-pyrazole-4-yl)-7-((R)-2-hydroxypropyl)-7H-pyrrolo[2,3-d]pyrimidine-5-yl)((S)-6-fluorochroman-3-yl)methanone; (2-(5-fluoro-1H-pyrazole-4-yl)-7-((R)-2-hydroxypropyl)-7H-pyrrolo[2,3-d]pyrimidine-5-yl)((S)-6-fluorochroman-3-yl)methanone hydrochloride; (2-(5-fluoro-1H-pyrazole-4-yl)-7-((R)-2-hydroxypropyl)-7H-pyrrolo[2,3-d]pyrimidine-5-yl)((S)-6-fluorochroman-3-yl)methanomethanesulfonic acid; (6-Fluorochroman-3-yl)(7-(2-hydroxy-2-methylpropyl)-2-(5-methoxy-1H-pyrazole-4-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-yl)methanone; (6-chlorochroman-3-yl)-[7-[(2S)-2-hydroxypropyl]-2-(5-methoxy-1H-pyrazole-4-yl)pyrrolo[2,3-d]pyrimidine-5-yl]methanone; (6-chlorochroman-3-yl)-[2-(5-fluoro-1H-pyrazole-4-yl)-7-(2-hydroxy-2-methylpropyl)pyrrolo[2,3-d]pyrimidine-5-yl]methanone; (S)-(6-chlorochroman-3-yl)(2-(5-fluoro-1H-pyrazole-4-yl)-7-(2-hydroxy-2-methylpropyl)-7H-pyrrolo[2,3-d]pyrimidine-5-yl)methanone; (S)-(6-chlorochroman-3-yl)(2-(5-fluoro-1H-pyrazole-4-yl)-7-(2-hydroxy-2-methylpropyl)-7H-pyrrolo[2,3-d]pyrimidine-5-yl)methanomethanesulfonic acid; (6-chlorochroman-3-yl)-[2-(5-chloro-1H-pyrazole-4-yl)-7-(2-hydroxy-2-methylpropyl)pyrrolo[2,3-d]pyrimidine-5-yl]methanone; (6-chlorochroman-3-yl)-[7-(2-hydroxy-2-methyl-propyl)-2-(5-methoxy-1H-pyrazole-4-yl)pyrrolo[2,3-d]pyrimidine-5-yl]methanone; (6-chlorochroman-3-yl)(1-(2-hydroxyethyl)-6-(5-methoxy-1H-pyrazole-4-yl)-1H-pyrrolo[3,2-c]pyridine-3-yl)methanone hydrochloride; (6-chlorochroman-3-yl)(1-(2-hydroxyethyl)-6-(5-methoxy-1H-pyrazole-4-yl)-1H-pyrrolo[3,2-c]pyridine-3-yl)methanomethanesulfonic acid; (2,3-dihydrobenzo[b][1,4]dioxin-2-yl)(1-(2-hydroxyethyl)-6-(1H-pyrazole-4-yl)-1H-indole-3-yl)methanone; Croman-3-yl-[1-(2-hydroxyethyl)-6-(1H-pyrazole-4-yl)indole-3-yl]methanone; 2,3-dihydro-1,4-benzodioxin-3-yl-[1-(2-hydroxyethyl)-6-(1H-pyrrolo[2,3-b]pyridine-3-yl)indole-3-yl]methanone; [6-(3-chloro-1H-pyrazole-4-yl)-1-(2-hydroxyethyl)indole-3-yl]-(2,3-dihydro-1,4-benzodioxin-3-yl)methanone; Chroman-3-yl-[1-(2-hydroxyethyl)-6-(1H-pyrrolo[2,3-b]pyridine-3-yl)indole-3-yl]methanone; [6-(3-chloro-1H-pyrazole-4-yl)-1-(2-hydroxyethyl)indole-3-yl]-chroman-3-yl-methanone; (6-Fluorochroman-3-yl)-[1-(2-hydroxyethyl)-6-(1H-pyrazole-4-yl)indole-3-yl]methanone; [1-(2-hydroxyethyl)-6-(1H-pyrazole-4-yl)indole-3-yl]-(7-methoxychroman-3-yl)methanone; (6-fluoro-2,3-dihydro-1,4-benzodioxin-3-yl)-[1-(2-hydroxyethyl)-6-(1H-pyrazole-4-yl)indole-3-yl]methanone; (6-chloro-2,3-dihydro-1,4-benzodioxin-2-yl)-[1-(2-hydroxyethyl)-6-(1H-pyrazole-4-yl)indole-3-yl]methanone; (6-fluoro-2,3-dihydro-1,4-benzodioxin-2-yl)-[1-(2-hydroxyethyl)-6-(1H-pyrazole-4-yl)indole-3-yl]methanone; (6,7-dichloro-2,3-dihydro-1,4-benzodioxin-3-yl)-[1-(2-hydroxyethyl)-6-(1H-pyrazole-4-yl)indole-3-yl]methanone; (6-chloro-2,3-dihydro-1,4-benzodioxin-3-yl)-[1-(2-hydroxyethyl)-6-(1H-pyrazole-4-yl)indole-3-yl]methaneone; [1-(2-hydroxyethyl)-6-(1H-pyrazole-4-yl)indole-3-yl]-(6-methoxy-2,3-dihydro-1,4-benzodioxin-3-yl)methaneone; [6-(3-chloro-1H-pyrazole-4-yl)-1-(2-hydroxyethyl)indole-3-yl]-(6-methoxychroman-3-yl)methanone; (6-chlorochroman-3-yl)(6-(3-fluoro-1H-pyrazole-4-yl)-1-(2-hydroxyethyl)-1H-indole-3-yl)methanone; (6-chlorochroman-3-yl)(1-(2-hydroxyethyl)-6-(3-methoxy-1H-pyrazole-4-yl)-1H-indole-3-yl)methanone; (S)-(6-chlorochroman-3-yl)(1-(2-hydroxyethyl)-6-(3-methoxy-1H-pyrazole-4-yl)-1H-indole-3-yl)methanone; (R)-(6-chlorochroman-3-yl)(1-(2-hydroxyethyl)-6-(3-methoxy-1H-pyrazole-4-yl)-1H-indole-3-yl)methanone; (6-Fluorochroman-3-yl)(1-(2-hydroxyethyl)-6-(3-methoxy-1H-pyrazole-4-yl)-1H-indole-3-yl)methanone; (S)-(6-fluorochroman-3-yl)(1-(2-hydroxyethyl)-6-(3-methoxy-1H-pyrazole-4-yl)-1H-indole-3-yl)methanone; (R)-(6-fluorochroman-3-yl)(1-(2-hydroxyethyl)-6-(3-methoxy-1H-pyrazole-4-yl)-1H-indole-3-yl)methanone; Croman-3-yl(1-(2-hydroxyethyl)-6-(1H-pyrazole-4-yl)-1H-indole-3-yl)methanone; Croman-3-yl(1-(2-methoxyethyl)-6-(1H-pyrazole-4-yl)-1H-indazole-3-yl)methanone; (S)-(2,3-dihydrobenzo[b][1,4]dioxin-2-yl)(1-(2-methoxyethyl)-6-(1H-pyrazole-4-yl)-1H-indazole-3-yl)methanone; (6-Fluorochroman-3-yl)(1-(2-hydroxyethyl)-6-(3-(trifluoromethyl)-1H-pyrazole-4-yl)-1H-indazole-3-yl)methanone; (6-Fluorochroman-3-yl)(1-(2-hydroxyethyl)-6-(3-methoxy-1H-pyrazole-4-yl)-1H-indazole-3-yl)methanone; (6-Fluorochroman-3-yl)(1-(2-hydroxyethyl)-6-(3-isopropyl-1H-pyrazole-4-yl)-1H-indazole-3-yl)methanone; 2-(3-(6-chlorochroman-3-carbonyl)-6-(3-methoxy-1H-pyrazole-4-yl)-1H-indazole-1-yl)acetonitrile; (2,3-dihydrobenzofuran-2-yl)(1-(2-hydroxyethyl)-6-(3-methoxy-1H-pyrazole-4-yl)-1H-indazole-3-yl)methanone; (1-(2-hydroxyethyl)-6-(3-methoxy-1H-pyrazole-4-yl)-1H-indazole-3-yl)(2-methyl-2,3-dihydrobenzo[b][1,4]dioxin-2-yl)methanone; Benzofuran-3-yl(1-(2-hydroxyethyl)-6-(3-methoxy-1H-pyrazole-4-yl)-1H-indazole-3-yl)methanone; (6-chlorochroman-3-yl)(1-(2-hydroxypropyl)-6-(3-methoxy-1H-pyrazole-4-yl)-1H-indazole-3-yl)methanone; (6-chlorochroman-3-yl)(1-((R)-2-hydroxypropyl)-6-(3-methoxy-1H-pyrazole-4-yl)-1H-indazole-3-yl)methanone; (6-chlorochroman-3-yl)(1-((S)-2-hydroxypropyl)-6-(3-methoxy-1H-pyrazole-4-yl)-1H-indazole-3-yl)methanone; (6-Fluorochroman-3-yl)(1-(2-hydroxyethyl)-6-(1H-pyrazole-4-yl)-1H-indazole-3-yl)methanone; (Croman-3-yl)(1-(2-hydroxyethyl)-6-(1H-pyrazole-4-yl)-1H-indazole-3-yl)methanone; (7-Fluoro-2,3-dihydrobenzo[b][1,4]dioxin-2-yl)(1-(2-hydroxyethyl)-6-(1H-pyrazole-4-yl)-1H-indazole-3-yl)methanone; and (7-chloro-2,3-dihydrobenzo[b][1,4]dioxin-2-yl)(1-(2-hydroxyethyl)-6-(1H-pyrazole-4-yl)-1H-indazole-3-yl)methanone.

[0079] (Examples) The present invention is further embodied by the following examples illustrating the preparation method according to the present invention of the compound shown in Figure 1. These examples are for illustrative purposes only and are not intended to limit the present invention in any way, nor should they be interpreted as limiting it. Those skilled in the art will understand that modifications and changes can be made without altering the scope of the present invention.

[0080] The nuclear magnetic resonance (NMR) and mass spectrometry (MS) spectra obtained for the following examples and the compounds described herein were consistent with the spectra of compounds having the formulas described herein.

[0081] Liquid chromatography-mass spectrometry (LC-MS) method: 1. The sample was collected using an Agilent Zorbax Eclipse XDB-C18 (3.5 μm) reverse-phase column (4.6 × 50 mm). The analysis was performed using the Technologies 6120 MSD system at room temperature with a flow rate of 1.5 mL / min. 2. The mobile phase consisted of solvent A (water / 0.1% formic acid) and solvent B (acetonitrile / 0.1% formic acid), which were used for 5 minutes, ranging from 95% / 5% to 0% / 100% (A / B). 1. The mass spectrum (m / z) was recorded using electrospray ionization (ESI). 2. Ionization data was rounded to the nearest integer.

[0082] Proton NMR spectrum: Unless otherwise specified, all 1 ¹H NMR spectra were measured using a Varian series Mercury 400 or 500 MHz. All observed protons were reported as ppm values ​​from tetramethylsilane to the lower field side, using abbreviations commonly used to denote major peaks, e.g., s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), brs (broad singlet).

[0083] (Example 1: Preparation of the compound) [ka]

[0084] Compound 1-1: [ka]

[0085] Step 1: A mixture of 6-chlorochroman-3-carboxylic acid (6.0 g, 28.2 mmol), thionyl chloride (20.98 mL, 282.18 mmol), and DMF (0.1 mL, catalyst) was heated under reflux in toluene (60 mL) with stirring for 3 hours. The mixture was cooled to room temperature and concentrated to remove volatile organic solvents. The crude product was dissolved in acetonitrile (100 mL) and concentrated to obtain 1a as a light brown oily substance (crude product). This was used in the next step without purification.

[0086] Step 2: A mixture of 1a (5.4 g, 23 mmol) and 6-bromo-1H-pyrrolo[3,2-c]pyridine (3.7 g, 19 mmol) dissolved in dichloromethane (60 mL) was cooled in an ice bath, and aluminum chloride (9.4 g, 70 mmol) was added. The mixture was stirred at 0°C for 1 hour. The mixture was quenched at 0°C with saturated NH4Cl solution (100 mL) to produce a solid precipitate. The mixture was then warmed to room temperature and stirred for a further 2 hours. The solid was collected by filtration and washed with methanol. The obtained solid was dried under high vacuum to obtain 1b as a white solid in 80% yield (7.4 g). LC / MS yielded 391.0 and 393.0 [M+H] + This was shown.

[0087] Step 3: A mixture of 1b (800 mg, 2.1 mmol), potassium carbonate (1.76 g, 1.3 mmol, 770 μL), and 2-chloro-N,N-dimethylethanamine hydrochloride (690 mg, 6.4 mmol) was heated to 70°C in DMF (20 mL) and stirred overnight. The mixture was poured into water (100 mL) and extracted with RINKAN (100 mL). The organic layer was separated, washed with saturated brine (100 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by flash column chromatography using a silica gel column (20-100% RINKAN / hexane gradient elution) to obtain a white solid 1c (202 mg) in 20% yield. LC / MS measurements yielded 463.1 and 465.0 [M+H]. + This was shown.

[0088] Step 4: Intermediate 1c (202 mg, 430 μmol), [3-methoxy-1-(p-tolylsulfonyl)pyrazole-4-yl]boronic acid (190 mg, 650 μmol), potassium carbonate (2 M aqueous solution, 540 μL), and tetrakis(triphenylphosphine)palladium (25.0 mg, 22 μmol) were dissolved in 1,4-dioxane (10 mL), degassed with argon gas for 5 minutes, and the reaction mixture was heated to 70°C and shaken overnight. The mixture was cooled to room temperature, concentrated, poured into water (50 mL), and extracted twice with ELISA (50 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated to obtain 1d, which was used in the next step without purification.

[0089] Step 5a:1d (crude product, 230 mg) of the tosyl (Ts) protected pyrazole compound was dissolved in THF (1 mL), cooled on an ice bath, and treated with 2N sodium hydroxide solution (2.0 mL). The solution was stirred at 0°C for 30 minutes, neutralized with 1N hydrochloric acid solution (4.0 mL), and diluted with  (30 mL) and saturated Na2CO3 solution (30 mL). The aqueous phase was separated and extracted with Â. The organic layers were combined, dried over Na2SO4, filtered, treated with silica gel, and evaporated under reduced pressure. The resulting substance was purified by silica gel column chromatography to obtain compound 1-1(6-chlorochroman-3-yl)-[1-[2-(dimethylamino)ethyl]-6-(3-methoxy-1H-pyrazole-4-yl)pyrrolo[3,2-c]pyridine-3-yl]methanone (101 mg, 210 μmol, 58% yield of steps 4 and 5a) as a white solid. The LC / MS result was 480.2 [M+H] + That was the case.

[0090] Step 6a: To a solution of ((6-chlorochroman-3-yl)(1-(2-(dimethylamino)ethyl)-6-(5-methoxy-1H-pyrazole-4-yl)-1H-pyrrolo[3,2-c]pyridine-3-yl)methanone hydrochloride):1-1 (50 mg, 104 mmol) in acetone (1 mL), 4 M HCl dioxane solution (55 mL) was slowly added at room temperature to form a solid precipitate. After stirring for a further 1 hour at room temperature, ter-butyl ethyl ether (2 mL) was added to completely precipitate the mixture. The solid was collected by filtration under a nitrogen atmosphere, washed with ter-butyl ethyl ether, and dried under reduced pressure to obtain 1-59 (47.5 mg, 82%) as a white powder. The LC / MS result was 480.2 [M+H]. + That was the case.

[0091] Step 6a: To a 1 mL solution of ((6-chlorochroman-3-yl)(1-(2-(dimethylamino)ethyl)-6-(5-methoxy-1H-pyrazole-4-yl)-1H-pyrrolo[3,2-c]pyridine-3-yl)methanomethanesulfonic acid):1-1 (50 mg, 104 mmol) in acetone (1 mL), a 220 mL solution of 1 M methanesulfonic acid was slowly added at room temperature to form a solid precipitate. After stirring for a further 1 hour at room temperature, 2 mL of ter-butyl ethyl ether was added to obtain a complete precipitate. The solid was collected by filtration under a nitrogen atmosphere, washed with ter-butyl ethyl ether, and dried under reduced pressure to obtain 1-60 (60.3 mg, 90%) as a white powder. The LC / MS result was 480.2 [M + H]. + That was the case.

[0092] Step 5b of the Boc-protected pyrazole compound: Crude tert-butyl 3-chloro-4-(3-(6-chloro-2H-chromen-3-carbonyl)-1-(2-hydroxyethyl)-1H-pyrrolo[3,2-c]pyridine-6-yl)-1H-pyrazole-1-carboxylate (40.7 mg) was dissolved in DCM (1.0 mL), and the solution was treated with TFA (0.5 mL) and stirred at room temperature. After stirring at room temperature for 2 hours, the reaction mixture was concentrated under reduced pressure to obtain the residue. The residue was dissolved in toluene (10 mL) and washed with 2 M Na2CO3 solution (10 mL). The aqueous layer was separated and extracted with toluene (5 mL), and the combined organic matter was dried over Na2SO4, filtered, treated with silica gel, and evaporated under reduced pressure. The obtained substance was purified by silica gel column chromatography to obtain compound 1-1(6-chloro-2H-chromen-3-yl)-[6-(3-chloro-1H-pyrazole-4-yl)-1-(2-hydroxyethyl)pyrrolo[3,2-c]pyridine-3-yl]methanone (33.0 mg, 41.8 μmol; 55% yield from steps 4 and 5b) as a light brown solid. The LC / MS result was 458.1 [M+H]. + That was the case.

[0093] The following compounds 1-2 to 1-60 shown in Tables 1-1 to 1-9 are suitable boronic acid / ester (R 2 ), and appropriate R 1 Compound 1-1 was prepared using a suitable substituent (4) by the same method (general formula 1) as described for the preparation of compound 1-1.

[0094] [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Table 1-10] [Table 1-11] [Table 1-12] [Table 1-13] [Table 1-14]

[0095] General formula 2 [ka] Compound 2-1: 2,3-dihydro-1,4-benzodioxin-3-yl-[1-(2-hydroxyethyl)-6-(1H-pyrazole-4-yl)indole-3-yl]methanone

[0096] formula 2A [ka]

[0097] Step 1: A DMSO (20 mL) solution of 6-bromo-1H-indole (2.00 g, 10.20 mmol) was treated with pulverized potassium hydroxide (865 mg, 15.42 mmol) and stirred at room temperature for 50 minutes. A DMSO (2.0 mL) solution of 2-bromoethanol (1.53 g, 12.24 mmol) was added dropwise (for 5 minutes) and stirred for 2 days. The solution was diluted with RINKAN (400 mL), washed sequentially with water (3 × 400 mL) and saturated saline (400 mL), dried (Na₂SO₄), filtered, and evaporated under reduced pressure. The residue was purified by silica gel column chromatography to obtain 2a, 2-(6-bromoindole-1-yl)ethanol as an oily substance (1.78 g, 7.41 mmol, yield 73%). LC / MS results were 240.1, +242.1 [M + H]. + That was the case.

[0098] Step 2: A solution of 2-(6-bromoindole-1-yl)ethanol (2a, 1.78 g, 7.41 mmol) in DMF (70 mL) was cooled in an ice bath and treated with N-iodosuccinimide (NIS, 1.75 g, 7.78 mmol). The ice bath was removed and the mixture was stirred at room temperature for 90 minutes. The solution was diluted with siRNA (500 mL), washed sequentially with water (3 × 500 mL) and saturated saline (500 mL), dried (Na₂SO₄), filtered, treated with silica gel, and evaporated under reduced pressure. The resulting substance was purified by silica gel column chromatography to obtain 2b, 2-(6-bromo-3-iodoindole-1-yl)ethanol (2.17 g, 5.93 mmol, yield 80%) as a pale yellow solid. LC / MS results were 365.0, 367.0 [M+H]. + That was the case.

[0099] Step 3: 2-(6-bromo-3-iodoindol-1-yl)ethanol (2b, 249.9 mg, 682.8 μmol) was placed in a 40 mL vial under a nitrogen atmosphere, and THF (9 mL) was added. The solution was cooled in a dry ice / acetonitrile bath (approximately -42°C), and isopropylmagnesium chloride (2 M THF solution, 0.72 mL) was added dropwise (for 2 minutes). After 2 minutes, the acetonitrile bath was replaced with an ice bath, and the sample was stirred for 30 minutes. The reaction mixture was cooled again to -42°C, and a solution of N-methoxy-N-methyl-2,3-dihydro-1,4-benzodioxin-3-carboxamide (4-2,183.6 mg, 822.5 μmol) dissolved in THF (2.5 mL) was added dropwise (for 3 minutes). After stirring the reaction mixture for 2 hours, it was removed from the cold bath, and the reaction was stopped by adding 12% NH4Cl aqueous solution (2 mL). THF was removed under reduced pressure. The aqueous residue was diluted to 20 mL with aqueous ammonium hydroxide solution and extracted with siRNA (40 mL). The organic layer was washed with saturated brine (40 mL), dried (Na₂SO₄), and evaporated under reduced pressure. The residue was purified by silica gel column chromatography to obtain the target [6-bromo-1-(2-hydroxyethyl)indole-3-yl]-(2,3-dihydro-1,4-benzodioxin-3-yl)methanone (2c, 56.6 mg, 140.7 μmol, yield 21%) as an oil. LC / MS results were 402.1, 404.1 [M+H]. + That was the case.

[0100] Step 4: [6-bromo-1-(2-hydroxyethyl)indole-3-yl]-(2,3-dihydro-1,4-benzodioxin-3-yl)methanone (2c, 53.4 mg, 132.8 μmol), tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole-1-carboxylate (60.7 mg, 206.4 μmol), PdCl2(dppf)-CH2Cl2 (16.6 mg, 20.33 μmol), and tribasic potassium phosphate (88.6 mg, 417.4 μmol) were dissolved in dioxane (2 mL) and water (0.24 mL). The suspension was bubbling with nitrogen for 6 minutes, and then shaken overnight at 65°C. The solution was evaporated under reduced pressure, the residue was mixed with water (10 mL), and extracted with butyl (3 × 10 mL). The combined organic matter was dried (Na2SO4), filtered, and evaporated under reduced pressure to obtain tert-butyl 4-[3-(2,3-dihydro-1,4-benzodioxin-3-carbonyl)-1-(2-hydroxyethyl)indole-6-yl]pyrazole-1-carboxylate (2d, 95.8 mg). The residue was used without further purification. The LC / MS result was 490.3 [M+H]. + That was the case.

[0101] Step 5: A solution of crude tert-butyl 4-[3-(2,3-dihydro-1,4-benzodioxin-3-carbonyl)-1-(2-hydroxyethyl)indole-6-yl]pyrazole-1-carboxylate (2d, 95.8 mg) in DCM (1 mL) and methanol (0.05 mL) was treated with TFA (0.5 mL) and shaken for 15 minutes. The solvent was evaporated under reduced pressure. The residue was dissolved in methanol (10 mL) and evaporated under reduced pressure. The residue was dissolved in DCM (10 mL), washed sequentially with ammonium hydroxide (1 M) and saturated saline (10 mL each), dried (Na2SO4), treated with silica gel, and evaporated under reduced pressure. The residue was purified by silica gel column chromatography to obtain 2,3-dihydro-1,4-benzodioxin-3-yl-[1-(2-hydroxyethyl)-6-(1H-pyrazole-4-yl)indole-3-yl]methanone 2-1 (16.2 mg, 41.6 μmol; 31% yield in steps 4 and 5) as an orange solid. LC / MS results were 390.2 [M+H] + That was the case.

[0102] Compound 2-2: (6-chlorochroman-3-yl)-[1-(2-hydroxyethyl)-6-(1H-pyrazole-4-yl)indole-3-yl]methanone

[0103] [ka]

[0104] Step 1: A solution of 2-(6-bromo-3-iodoindol-1-yl)ethanol (2b, 598.1 mg, 1.63 mmol), imidazole (225.0 mg, 3.31 mmol), and DMAP (6.7 mg, 54.84 μmol) in DMF (4 mL) was treated with TBSCl (397.6 mg, 2.64 mmol) and stirred at room temperature for 16 hours. The solution was diluted with  (40 mL), washed sequentially with water and saturated saline (40 mL each), dried (Na2SO4), treated with silica gel, and evaporated under reduced pressure. The residue was purified by silica gel column chromatography to obtain 2-(6-bromo-3-iodoindol-1-yl)ethoxy-tert-butyldimethylsilane (2f, 0.63 g, 1.31 mmol, yield 80%). The LC / MS results were 479.9, 481.9 [M+H]. + That was the case.

[0105] Step 2: A solution of 2-(6-bromo-3-iodoindol-1-yl)ethoxy-tert-butyldimethylsilane (2f, 449.2 mg, 935.3 μmol) in anhydrous THF (13 mL) was cooled in a dry ice / acetonitrile bath (approximately -42°C), and isopropylmagnesium chloride (2 M THF solution, 0.65 mL) was added dropwise (for 3 minutes). After 3 minutes, the acetonitrile bath was replaced with an ice bath, and the reaction mixture was stirred for 45 minutes. The reaction mixture was cooled again to -42°C, and a solution of 6-chloro-N-methoxy-N-methylchroman-3-carboxamide (4-1,363.8 mg, 1.42 mmol) in THF (3.6 mL) was added dropwise (for 3 minutes). After 3 minutes, the dry ice bath was replaced with an ice water bath, and the mixture was stirred for 2.5 hours. The reaction was stopped with water (2 mL), and the THF was evaporated under reduced pressure. The residual aqueous solution was diluted with toluene (40 mL), washed sequentially with water and saturated saline (40 mL each), dried (Na₂SO₄), filtered, treated with silica gel, and evaporated under reduced pressure. The residue was purified by silica gel column chromatography to obtain [6-bromo-1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]indole-3-yl]-(6-chlorochroman-3-yl)methanone (2 g, 194.5 mg, 354.30 μmol, yield 38%) as a clear yellow oil. LC / MS results were 548.3, 550.3 [M+H]. + That was the case.

[0106] Step 3: [6-bromo-1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]indole-3-yl]-(6-chlorochroman-3-yl)methanone (2 g, 63.9 mg, 116.4 μmol), tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole-1-carboxylate (52.4 mg, 178.1 μmol), PdCl2(dppf)-CH2Cl2 (15.1 mg, 18.5 μmol), and tribasic potassium phosphate (79.0 mg, 372.2 μmol) were dissolved in dioxane (2.1 mL) and water (0.25 mL). The suspension was bubbling with nitrogen for 5 minutes, and then shaken overnight at 65°C. The solution was evaporated under reduced pressure, the residue was dissolved in toluene (10 mL), and washed sequentially with water and saturated saline (10 mL each). After drying (Na2SO4) and filtering, the mixture was evaporated under reduced pressure to obtain crude tert-butyl 4-[1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-3-(6-chlorochroman-3-carbonyl)indole-6-yl]pyrazole-1-carboxylate (2h, 99.3 mg) as a red oil. The residue was used without purification. The LC / MS result was 636.2[M+H]. + That was the case.

[0107] Step 4: A solution of crude tert-butyl 4-[1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-3-(6-chlorochroman-3-carbonyl)indole-6-yl]pyrazole-1-carboxylate (2h, 99.3 mg) in DCM (1.0 mL) and MeOH (0.05 mL) was treated with TFA (1.0 mL) and stirred at room temperature. After about 2 hours, the solvent was evaporated under reduced pressure to obtain a red residue. This substance was dissolved in DCM (10 mL) and washed with 1 M ammonium hydroxide. The aqueous layer was extracted with DCM (10 mL), and the combined organic layer was treated with methanol to obtain a clear red solution. This solution was treated with silica gel and evaporated under reduced pressure. The residue was purified by silica gel column chromatography to obtain (6-chlorochroman-3-yl)-[1-(2-hydroxyethyl)-6-(1H-pyrazole-4-yl)indole-3-yl]methanone (2-2, 26.8 mg, 63.5 μmol; 55% yield in steps 3 and 4) as a brown solid. LC / MS results were 422.2 [M+H]. + That was the case.

[0108] Compound 2-24: Croman-3-yl-[1-(2-methoxyethyl)-6-(1H-pyrazole-4-yl)indazole-3-yl]methanone

[0109] [ka]

[0110] Step 1: A suspension of 6-bromo-3-iodo-1H-indazole (2.00 g, 6.19 mmol) and potassium carbonate (2.57 g, 18.58 mmol) in DMF (30 mL) was stirred at room temperature for 75 minutes. 1-chloro-2-methoxyethane (0.70 mL, 7.70 mmol) was added, and the suspension was stirred at 60°C for 9 hours. The mixture was diluted with siRNA (225 mL), sequentially washed with water (3 × 225 mL) and saturated saline (225 mL), dried (Na₂SO₄), filtered, and evaporated under reduced pressure. The residue was purified by silica gel column chromatography to obtain 6-bromo-3-iodo-1-(2-methoxyethyl)indazole (3a, 1.70 g, 4.47 mmol, yield 72%) as a fluffy white solid. LC / MS results were 381.0, 383.0 [M+H]. + That was the case.

[0111] Step 2: To a solution of 6-bromo-3-iodo-1-(2-methoxyethyl)indazole (3a, 485.8 mg, 1.3 mmol) in THF (20 mL), isopropyl magnesium chloride (2 M THF solution, 0.76 mL) was added dropwise at -78 °C. After stirring at -78 °C for 35 minutes, a solution of N-methoxy-N-methylchroman-3-carboxamide (4-3,443.2 mg, 2.00 mmol) in THF (5 mL) was added dropwise. After 1 hour and 45 minutes, water (3 mL) was added dropwise to stop the reaction, and the mixture was concentrated under reduced pressure. The residue was diluted with  (40 mL) and washed sequentially with aqueous ammonium chloride (12%, 20 mL) and saturated saline (40 mL). The organic layer was dried (Na₂SO₄), separated, and then evaporated under reduced pressure. The residue was purified by silica gel column chromatography to obtain [6-bromo-1-(2-methoxyethyl)indazole-3-yl]chroman-3-ylmethanone (3b, 50.7 mg, 122.1 μmol, yield 10%) as an oil. LC / MS results were 415.1,417.1[M+H]. + That was the case.

[0112] Step 3: A suspension of dioxane (1.85 mL) and water (0.21 mL) containing [6-bromo-1-(2-methoxyethyl)indazole-3-yl]-chroman-3-yl-methanone (3b, 50.7 mg, 122.1 μmol), tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole-1-carboxylate (71.8 mg, 244.1 μmol), PdCl2(dppf)-CH2Cl2 (15.2 mg, 18.61 μmol), and tribasic potassium phosphate (79.1 mg, 372.64 μmol) was bubbling with nitrogen for 5 minutes and shaken overnight at 65°C. The residue was dissolved in Â(10 mL), washed with saturated saline (10 mL), dried (Na₂SO₄), filtered, and evaporated under reduced pressure to obtain impure tert-butyl 4-[3-(chroman-3-carbonyl)-1-(2-methoxyethyl)indazole-6-yl]pyrazole-1-carboxylate (3c, 119.6 mg). This substance was used without further purification. The LC / MS result was 503.2[M+H]. + That was the case.

[0113] Step 4: Crude tert-butyl 4-[3-(chroman-3-carbonyl)-1-(2-methoxyethyl)indazole-6-yl]pyrazole-1-carboxylate (3c, 119.6 mg) was treated with DCM (1.0 mL) and MeOH (0.05 mL) solutions and shaken with TFA (0.5 mL) for 15 minutes. The solvent was evaporated under reduced pressure. The residue was dissolved in  (10 mL), washed sequentially with 1 M NH4OH and saturated saline (10 mL each), dried (Na2SO4), separated, treated with silica gel, and evaporated under reduced pressure. The residue was purified by silica gel column chromatography to obtain chroman-3-yl-[1-(2-methoxyethyl)-6-(1H-pyrazole-4-yl)indazole-3-yl]methanone (2-24, 28.3 mg, 70.32 μmol; 58% yield from Steps 3 and 4) as a pink solid. The LC / MS result was 403.2[M+H] + That was the case.

[0114] Compounds 2-3 to 2-35, shown in Table 2, were prepared using appropriate boronic acid / ester and winelevamide in the same manner as described for the preparation of compounds 2-1, 2-2, or 2-24.

[0115] [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6] [Table 2-7] [Table 2-8] [Table 2-9] [Table 2-10] [Table 2-11] [Table 2-12]

[0116] Alternatively, a racemic mixture of compounds was separated by the following supercritical fluid chromatography.

[0117] Compound 2-18: [(3S)-6-chlorochroman-3-yl]-[6-(5-methoxy-1H-pyrazole-4-yl)-1-(2-oxydanylethyl)indole-3-yl]methanone and Compound 2-19: [(3R)-6-chlorochroman-3-yl]-[6-(5-methoxy-1H-pyrazole-4-yl)-1-(2-oxydanylethyl)indole-3-yl]methanone

[0118] [ka]

[0119] A sample of racemic compound 2-17 was separated by supercritical fluid chromatography under the following conditions: Column: ChiralPak IC-H 21x250mm; Mobile phase: 40% methanol (CO2 solution); Flow rate: 70 mL / min; Sample: 19.4 mg dissolved in 2.0 mL of methanol and 2.0 mL of dichloromethane; Injection volume: 0.75 mL; Detection wavelength: 220 nm.

[0120] Compound 2-18: [(3S)-6-chlorochroman-3-yl]-[6-(5-methoxy-1H-pyrazole-4-yl)-1-(2-oxydanylethyl)indole-3-yl]methanone: First elution peak: 8.4 mg, purity 100%, 97.9% ee. LC / MS result: 452.2 [M+H] + That was the case.

[0121] Compound 2-19: [(3R)-6-chlorochroman-3-yl]-[6-(5-methoxy-1H-pyrazole-4-yl)-1-(2-oxydanylethyl)indole-3-yl]methanone: Second elution peak: 8.7 mg, purity 100%, 96.4% ee. LC / MS result: 452.2 [M+H] + That was the case.

[0122] Compound 2-21: [(3S)-6-fluorochroman-3-yl]-[1-(2-hydroxyethyl)-6-(5-methoxy-1H-pyrazole-4-yl)indole-3-yl]methanone and Compound 2-22: [(3R)-6-fluorochroman-3-yl]-[1-(2-hydroxyethyl)-6-(5-methoxy-1H-pyrazole-4-yl)indole-3-yl]methanone

[0123] [ka]

[0124] A sample of racemic compound 2-20 was separated by supercritical fluid chromatography under the following conditions: Column: ChiralPak IC-H 21 x 250 mm; Mobile phase: 35% methanol (CO2); Flow rate: 70 mL / min; Sample: 25.5 mg dissolved in 2.0 mL of methanol and 2.0 mL of dichloromethane; Injection volume: 1.0 mL; Detection wavelength: 254 nm.

[0125] Compound 2-21: [(3S)-6-fluorochroman-3-yl]-[1-(2-hydroxyethyl)-6-(5-methoxy-1H-pyrazole-4-yl)indole-3-yl]methanone: First elution peak, 10.9 mg, 100% purity, 98.5% ee. LC / MS result: 436.2 [M+H] + That was the case.

[0126] Compound 2-22: [(3R)-6-fluorochroman-3-yl]-[1-(2-hydroxyethyl)-6-(5-methoxy-1H-pyrazole-4-yl)indole-3-yl]methanone: Second elution peak: 10.8 mg, purity 100%, 97.7% ee. LC / MS result: 436.2 [M+H] + That was the case.

[0127] Compound 3-1: Croman-3-yl-[1-(2-methoxyethyl)-6-(1H-pyrazole-4-yl)indazole-3-yl]methanone

[0128] [ka]

[0129] Step 1: A mixture of 6-bromo-1H-indazole (985 mg, 5.00 mmol) and 1-tetrahydropyran-2-yl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole (2.09 g, 7.50 mmol), tris(dibenzylideneacetone)dipalladium, tri-t-butylphosphonium tetrafluoroborate (4 mol%), and Na2CO3 (3 equivalents) was dissolved in a 1:1 mixture of ethanol / water (10 mL), degassed by aeration with nitrogen d for 5 minutes, heated to 100°C, and stirred overnight. The mixture was cooled to room temperature, poured into water, and extracted with ethyl acetate. The combined organic layer was washed with saturated brine, dried over MgSO4, filtered, and concentrated. The residue was purified by silica gel column chromatography with 50-100% siRNA in heptane to obtain 4a,6-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-4-yl)-1H-indazole as a white foamy solid in 90% yield (4a, 1.21 g, 4.51 mmol). LC / MS results were 269.2 [M+H]. + That was the case.

[0130] Step 2: Potassium carbonate (1.04 g, 7.53 mmol) was added to a 10 mL solution of 6-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-4-yl)-1H-indazole (4a, 1.01 g, 3.76 mmol) in DMF (10 mL), followed by N-iodosuccinimide (1.02 g, 4.52 mmol). The mixture was stirred overnight at room temperature and then poured into a 10% NaHSO3 aqueous solution. The mixture was extracted with siRNA, the combined organic layer was washed with water and saturated brine, dried over Na2SO4, filtered, and concentrated to obtain 3-iodo-6-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-4-yl)-1H-indazole as a white solid in 86% yield (4b, 1.27 g, 3.22 mmol). LC / MS result: 395.1 [M+H]+ The crude product was used without further purification.

[0131] Step 3: Potassium carbonate (888 mg, 6.43 mmol) was added to a 32 mL solution of 3-iodo-6-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-4-yl)-1H-indazole (4b, 1.27 g, 3.22 mmol) in acetonitrile (32 mL), followed by the addition of ethylene oxide (2.9 M in THF, 2.2 mL, 6.4 mmol). The mixture was heated overnight at 80°C with stirring. The mixture was cooled to room temperature, poured into water, and extracted with siRNA. The combined organic layers were washed with saturated brine, dried over Na2SO4, filtered, and then concentrated. The residue was purified by silica gel column chromatography using a gradient of 50-100% siRNA in heptane to obtain 4c,2-(3-iodo-6-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-4-yl)-1H-indazole-1-yl)ethane-1-ol as a solid in 32% yield (450 mg, 1.03 mmol). LC / MS results were 439.1 [M+H]. + That was the case.

[0132] Step 4: A solution of 2-(3-iodo-6-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-4-yl)-1H-indazole-1-yl)ethane-1-ol (4c, 437 mg, 1.00 mmol) and imidazole (140 mg, 2.06 mmol) in DMF (4 mL) was cooled to 0°C, and tert-butyldimethylchlorosilane (170 mg, 1.13 mmol) was added. The mixture was warmed to room temperature and stirred overnight. The mixture was poured into water and extracted with 1:1 siRNA / hexane. The combined organic layers were washed with water (3 times), washed with saturated brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography using a gradient of 1-10% siRNA in hexane to obtain 1-(2-((tert-butyldimethylsilyl)oxy)ethyl)-3-iodo-6-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-4-yl)-1H-indazole as a white solid in 88% yield (4d, 487 mg, 0.88 mmol). LC / MS results were 553.3[M+H]. + That was the case.

[0133] Step 5: A solution of 1-(2-((tert-butyldimethylsilyl)oxy)ethyl)-3-iodo-6-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-4-yl)-1H-indazole (4d, 85 mg, 0.15 mmol) in dry THF (2 mL) was cooled to -40°C, and isopropyl magnesium chloride (2.0 M in THF, 0.11 mL, 0.22 mmol) was added dropwise. After 5 minutes, the mixture was warmed to 0°C and stirred at this temperature for 45 minutes. The mixture was cooled again to -40°C, and 6-fluoroanyl-N-methoxy-N-methylchroman-3-carboxamide (62 mg, 0.26 mmol) dissolved in dry THF (1 mL) was added dropwise (1 mL). The mixture was reheated to 0°C and stirred for 2 hours. The reaction was stopped with water, and the mixture was extracted with ELISA. The bonded organic layer was washed with a 10% aqueous ammonium hydroxide solution, then with saturated brine, dried over Na2SO4, filtered, concentrated, and dried. The residue was purified by silica gel column chromatography with a gradient of 1-10% siRNA in heptane to obtain (1-(2-((tert-butyldimethylsilyl)oxy)ethyl)-6-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-4-yl)-1H-indazole-3-yl)(6-fluorochroman-3-yl)methanone as a white solid in 38% yield (4e, 35 mg, 59 μmol).

[0134] Step 6: Add 11 mg of p-toluenesulfonic acid monohydrate to a 1:1 mixture (2 mL) of (1-(2-((tert-butyldimethylsilyl)oxy)ethyl)-6-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-4-yl)-1H-indazole-3-yl)(6-fluorochroman-3-yl)methanone (4e, 35 mg, 59 μmol) in THF / water, and stir the mixture overnight while heating it to 60°C. Cool the mixture to room temperature, pour it into water, and extract with ethyl acetate. Wash the combined organic layers with saturated brine, dry over Na2SO4, filter, and concentrate. The residue was purified by silica gel column chromatography using a gradient of 1-10% CH3OH in CH2Cl2, yielding (6-fluorochroman-3-yl)(1-(2-hydroxyethyl)-6-(1H-pyrazole-4-yl)-1H-indazole-3-yl)methanone as a white solid in 24% yield (3-1, 5.7 mg, 14 μmol). LC / MS results were 407.1 [M+H]. + That was the case.

[0135] Compounds 3-2 to 3-4 shown in Table 3 were prepared using the same method as for compound 3-1, with appropriate winerebamide and appropriate R 2 It was prepared using boronic acid esters.

[0136] [Table 3]

[0137] [ka] Intermediate 4-1:6-chloro-N-methoxy-N-methylchroman-3-carboxyamide [ka]

[0138] A suspension of 6-chlorochroman-3-carboxylic acid (964.5 mg, 4.54 mmol), N,O-dimethylhydroxylamine hydrochloride (665.3 mg, 6.82 mmol), and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate and N-[(dimethylamino)-1H-1,2,3-triazolo[4,5-b]pyridine-1-ylmethylene]-N-methylmethananium hexafluorophosphate N-oxide (HATU) (2.15 g, 5.67 mmol) in DMF (33 mL) was treated with N,N-diisopropylethylamine (18.4 mmol, 3.2 mL). The solution was stirred overnight at room temperature. The solution was diluted with toluene (300 mL), washed sequentially with water (3 × 300 mL) and saturated saline (300 mL), dried (Na₂SO₄), filtered, and evaporated under reduced pressure. The resulting substance was purified by silica gel column chromatography to obtain 6-chloro-N-methoxy-N-methylchroman-3-carboxamide (Weinlev intermediate 4-1; 1.06 g, 4.15 mmol, yield 91%) as a clear, colorless oil. The LC / MS result was 256.0 [M + H]. + That was the case.

[0139] The following wine rebuamide intermediates 4-2 to 4-23, shown in Table 4, were prepared using the same method as for the preparation of intermediate 4-1, with appropriate carboxylic acids.

[0140] [Table 4-1] [Table 4-2] [Table 4-3]

[0141] Example 2: ROCK Inhibition assay Kinase IC 50The activity was measured by an in vitro assay based on Perkin Elmer's LANCE Ultra TR-FRET (time-resolved fluorescence resonance energy transfer) homogeneous technique. Recombinant ROCK1 (amino acids 1-477) and ROCK2 (amino acids 5-554) proteins were purchased from Karna Biosciences and Signalchem. Compound activity was measured by Envision and IC50. 50 The following was calculated. The assay was performed on a white LUMITRAC® 2009 6-well half-area microplate from Greiner Bio One. The kinase reaction buffer consisted of 50 mM HEPES, pH 7.5, 1 mM EGTA, 10 mM MgCl2, 2 mM DTT, and 0.01% Tween-20. The kinase was incubated with 50 nM (ULight-CREBtide) substrate in the presence of 1 mM (ROCK1) or 1 mM (ROCK2) ATP. The kinase reaction was carried out for 1 hour, after which a reaction stop buffer was added, and the solution consisted of 10 mM EDTA and 0.6 nM LANCE UltraEuropium-labeled antiphosphorylated CREB (Ser133) antibody (PerkinElmerTRF0200) in LANCE detection buffer. All assay incubations were performed at room temperature, during which the microplates were sealed with polyester film. After incubating the reaction solution for 1 hour, the signal was measured using Envision in TR-FRET mode (excitation wavelength 320 nm, fluorescence wavelength 615 / 665 nm).

[0142] The compound of formula (1) exhibited useful pharmacological properties. In this specification, the method for expressing the inhibitory activity (nM) is the 50% inhibitory activity value (IC2). 50 ) The results are shown in Table 5 below. Here, IC 50 If it is less than 10 nM, it is designated as "A", IC 50 When the value is between 11nM and 50nM, it is designated as "B", IC 50 When the value is between 51nM and 500nM, it is designated as "C", and IC 50 A value exceeding 501 nM is defined as "D". Table 5 shows the inhibition of ROCK1 and ROCK2 by representative compounds of formula (1).

[0143] Table 5. Inhibitory activity of ROCK1 and ROCK2 in ATP Km values [Table 5-1] [Table 5-2]

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