ROCK kinase inhibitors

Abstract

This invention relates to compounds that inhibit rock activity. In particular, this invention relates to compounds, pharmaceutical compositions, and methods of use, such as methods for inhibiting rock activity and methods for treating, for example, cerebral cavernous syndrome (CCM) and cardiovascular diseases using the compounds and pharmaceutical compositions of this invention.

Description

Field of the Invention

[0001] The present invention relates to compounds that are Rho-associated protein kinase (ROCK) inhibitors. In particular, the present invention relates to compounds that selectively inhibit ROCK activity, pharmaceutical compositions containing the compounds, and methods of use thereof. Background Art

[0002] ROCK is an important RhoA effector that regulates multiple cellular signaling pathways through the phosphorylation of multiple substrates, including myosin light chain (MLC), LIM kinase, MYPT1, and CPI-17. ROCK substrates are involved in smooth muscle contractility, vascular permeability, actin filament organization, cell adhesion, cell migration, growth control, and cytokinesis. The direct involvement of ROCK in smooth muscle contraction is due to increased phosphorylation of MLC (which increases contractility after phosphorylation) and an increase in MLC phosphatases (which are inhibited by phosphorylation), which further increases MLC phosphorylation ROCK activation, resulting in increased contractility of smooth muscle.

[0003] Literature reports indicate that ROCK activity is associated with a number of cardiovascular diseases associated with vascular dysfunction, including hypertension, atherosclerosis, ischemic stroke, coronary artery spasm, cerebral vasospasm, angina pectoris, erectile dysfunction, and kidney disease, as well as other smooth muscle hyperreactivity such as glaucoma and asthma.

[0004] Brain cavernous malformation (CCM) syndrome is a disease associated with endothelial junction defects, primarily in the brain, that puts patients at lifelong risk for epilepsy, hemorrhagic stroke, and other neurological deficits. Brain cavernous malformations (CCMs) are vascular malformations found mostly within the central nervous system, such as the brainstem and spinal cord, and can occur in a sporadic or autosomal dominant manner, the latter mapping to three loci KRIT-1 / CCM1, MGC4607 / OSM / CCM2, and PDCD10 / CCM3 (e.g., see Glading et al., (2007) J. Cell Biol. 179: 247-254).

[0005] Each of the CCM1, CCM2, and CCM3 proteins interacts with a number of proteins, including Rap1, a RhoA-GTPase. Rap1 plays a role in regulating the actin cytoskeleton by stimulating the formation of actin fibers. Inactivating mutations in CCM1, CCM2, or CCM3 are sufficient to induce CCM vasculopathy by inhibiting endothelial cell vascular tube formation and invasion of the extracellular matrix, leading to weakened cell-cell connections and increased vascular leakage. Loss of expression of CCM1, -2, or -3 proteins results in significantly higher expression and increased activation of Rap1. Increased Rap1 activation is associated with increased Rho kinase-dependent (ROCK) phosphorylation and short hairpin RNA knockdown of ROCK that rescues endothelial cells from CCM pathology (e.g., see Borikova et al., (2010) J. Biol. Chem. 285: 11760-11764).

[0006] Unfortunately, patients with CCM have limited treatment options, and there are no approved drugs for the treatment of CCM in the United States. Fasudil hydrochloride, an isoquinoline sulfonamide derivative, is a potent ROCK inhibitor that has been approved outside the United States for the treatment of cerebral vasospasm following subarachnoid hemorrhage and symptomatic cerebral ischemia. Fasudil has been used off-label by physicians to treat CCM, presumably by inhibiting ROCK kinase activity; however, fasudil has poor selectivity for other AGC family kinases and also exhibits a number of serious toxic side effects and drawbacks, including mandatory short-course therapy, low oral bioavailability, cytotoxicity, and blood pressure fluctuations (e.g., see Xin et al., (2015) BioSci Rep. 35:1-13).

[0007] There is a significant unmet medical need for the treatment of patients with CCM, and more broadly, for the treatment of a number of other diseases involving elevated smooth muscle contractility, such as cardiovascular disease, with predominant evidence suggesting that selective ROCK kinase inhibitors may be a promising pharmacological therapeutic intervention. Coupled with this, there is a need to develop novel ROCK kinase inhibitors with improved duration of treatment, improved oral bioavailability, reduced cytotoxicity, and reduced off-target activity, particularly relative to AGC family kinase members. Summary of the invention

[0008] The present inventors have recognized the need to develop novel ROCK kinase inhibitors that exhibit improved cellular potency, kinome selectivity, efficacy, stability and safety. The compounds and compositions of the present invention advantageously overcome one or more of these disadvantages by providing potent, selective and orally active ROCK inhibitors.

[0009] In one aspect of the present invention, a compound that inhibits ROCK kinase activity is provided. In certain embodiments, the compound is represented by formula (Ia), formula (Ib), formula (IIa) or formula (IIb):

[0010]

[0011]

[0012] or a pharmaceutically acceptable salt thereof:

[0013] in:

[0014] X is a partially saturated nitrogen-containing heteroaryl group;

[0015] Y is N or CH;

[0016] Each R 1 is cyano, hydroxy, hydroxyalkyl, halogen, haloalkyl, alkoxy, C-C1-C3 alkyl, -N(R 4 )2, -NR 4 C(O)C1-C3 alkyl or cycloalkyl;

[0017] Q is a bond, O or N;

[0018] Each R 2 is hydrogen or halogen;

[0019] Each R 3 is hydrogen, halogen or C1-C3 alkyl;

[0020] Each R 4 is hydrogen or C1-C3 alkyl; and

[0021] m is 0, 1, 2 or 3.

[0022] In another aspect of the present invention, a pharmaceutical composition is provided, which comprises a therapeutically effective amount of a compound of Formula (Ia), Formula (Ib), Formula (IIa) or Formula (IIb) and a pharmaceutically acceptable excipient, salt, solvate, carrier and / or diluent.

[0023] In another aspect, the present invention relates to a method for treating cardiovascular diseases associated with increased vascular tone, the method comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula (Ia), formula (Ib), formula (IIa) or formula (IIb) or a pharmaceutical composition comprising the compound. In one embodiment, the cardiovascular disease is hypertension, atherosclerosis, ischemic stroke, coronary artery spasm, cerebral vasospasm, angina pectoris and erectile dysfunction.

[0024] On the other hand, the present invention relates to a method for treating a disease involving increased contractility of non-vascular smooth muscle, the method comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula (Ia), formula (Ib), formula (IIa) or formula (IIb) or a pharmaceutical composition comprising the compound. In one embodiment, the disease involves asthma and glaucoma with increased contractility of non-vascular smooth muscle.

[0025] In another aspect of the present invention, a method for inhibiting ROCK activity in a cell is provided, the method comprising contacting the cell with a compound of formula (Ia), (Ib), (IIa) or (IIb). A method for treating CCM in a patient is also provided, the method comprising administering a therapeutically effective amount of a compound of formula (Ia), (Ib), (IIa) or (IIb) or a pharmaceutical composition comprising the compound to a patient in need. DETAILED DESCRIPTION

[0026] The present invention relates to ROCK kinase inhibitors. In particular, the present invention relates to compounds that selectively inhibit ROCK kinase activity, pharmaceutical compositions comprising therapeutically effective amounts of the compounds, and methods of use thereof, such as methods for treating cerebral cavernous malformation syndrome (CCM) and cardiovascular disease.

[0027] definition

[0028] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the invention belongs. All patents, patent applications and publications cited herein are incorporated herein by reference to the extent consistent with the present disclosure. Unless otherwise expressly defined, terms and scopes are generally defined as such.

[0029] For simplicity, chemical groups are defined and are primarily referred to throughout the text as monovalent chemical groups (e.g., alkyl, aryl, etc.). However, these terms may also be used to convey corresponding multivalent groups in the context of appropriate structures that are clear to one skilled in the art. All atoms are understood to have normal bond valences (i.e., 4 for carbon, 3 for N, 2 for O, and 2, 4, or 6 for S, depending on the oxidation state of S).

[0030] As used herein, "ROCK kinase" refers to members of the Rho-associated protein kinase ("ROCK") family, including ROCK1 and ROCK2 kinases.

[0031] As used herein, "ROCK inhibitors" refer to compounds of the present invention represented by formula (I) or (II) as described herein. These compounds are capable of negatively regulating or inhibiting all or part of the enzymatic activity of ROCK kinase.

[0032] As used herein, the term "alkyl" refers to straight and branched chain aliphatic groups having 1 to 12 carbon atoms. Thus, "alkyl" encompasses C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 and C 12 Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, and hexyl.

[0033] The term "cyano" refers to -CN.

[0034] The term "cycloalkyl" as used herein refers to saturated and partially unsaturated cyclic hydrocarbon groups containing 3 to 12 carbon atoms. Thus, "cycloalkyl" includes C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 and C 12 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl.

[0035] As used herein, the term "heteroaryl" refers to a group having 5 to 14 ring atoms, preferably 5, 6, 10, 13 or 14 ring atoms; having 6, 10 or 14 pi electrons shared in a cyclic array; and having, in addition to carbon atoms, 1 to 3 heteroatoms each independently being N, O or S. "Heteroaryl" also includes fused polycyclic (e.g., bicyclic) ring systems in which one or more of the fused rings is non-aromatic, provided that at least one ring is aromatic and at least one ring contains N, O or S ring atoms.

[0036] Examples of heteroaryl groups include acridinyl, azoalkynyl, benzimidazolyl, benzofuranyl, benzo[d]oxazol-2(3H)-one, 2H-benzo[b][1,4]oxazin-3(4H)-one, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzo[d]oxazol-2(3H)-one, 2H-benzo[b][1,4]oxazin-3(4H)-one, olyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, romenyl, cinnolinyl, furanyl, furazanyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indazolyl, isobenzofuranyl, isochroman yl), isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,3-thiadiazolyl, 1,2, 4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl and xanthenyl.

[0037] The term "nitrogen-containing heteroheteroaryl" refers to a heteroaryl group containing nitrogen atoms in place of carbon atoms in a monocyclic, bicyclic, polycyclic or fused group having 5 to 14 ring atoms, preferably 5, 6, 10, 13 or 14 ring atoms; there are 6, 10 or 14 pi electrons in the cyclic array. "Nitrogen-containing heteroheteroaryl" also includes fused polycyclic (e.g., bicyclic) ring systems in which one or more of the fused rings is non-aromatic, provided that at least one ring is aromatic and at least one ring contains N.

[0038] Exemplary nitrogen-containing heteroaryl groups include, but are not limited to, indolizinyl, indolyl, isoindolyl, 3H-indolyl, indolyl, indazolyl, 1H-indazolyl, benzimidazolyl, benzothiazolyl, purinyl, 4H-quinolizinyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 1,8-naphthyridinyl, dinyl), pteridinyl, quinuclidine, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, 2H-pyrrolyl, pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, oxazolyl, thiazolyl, imidazolyl, 2-imidazolinyl, pyrazolyl, 2-pyrazolyl, isoxazolyl, isothiazolyl, 1,2,3-oxadiazolyl, 1,2,3-triazolyl, 1,3,4-thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl and 1,3,5-triazinyl.

[0039] The term "haloalkyl" refers to an alkyl chain in which one or more hydrogens have been replaced by a halogen. Exemplary haloalkyls are trifluoromethyl, difluoromethyl, fluorochloromethyl and fluoromethyl.

[0040] As used herein, the term "halogen" refers to chlorine, bromine, fluorine or iodine.

[0041] The term "hydroxy" refers to -OH.

[0042] The term "hydroxyalkyl" refers to an -alkyl-OH group.

[0043] As used herein, an "effective amount" of a compound is an amount sufficient to negatively regulate or inhibit the activity of ROCK kinase.

[0044] As used herein, a "therapeutically effective amount" of a compound is sufficient to improve or in some way alleviate symptoms or stop or reverse the process, such as CCM, or negatively regulate or inhibit the activity of ROCK kinase. Such an amount can be administered in a single dose or can be administered according to an effective regimen.

[0045] As used herein, "treat" or "treatment" refers to any manner in which the symptoms or pathology of a patient's condition, disorder or disease are ameliorated or otherwise beneficially altered.

[0046] As used herein, amelioration of the symptoms of a particular condition by administration of a particular compound or pharmaceutical composition refers to any relief, whether permanent or temporary, lasting or transient, attributable to or associated with administration of the composition.

[0047] Compound

[0048] In one aspect of the present invention, there is provided a compound represented by formula (Ia), formula (Ib), formula (IIa) or formula (IIb):

[0049]

[0050] or a pharmaceutically acceptable salt thereof:

[0051] in:

[0052] X is a partially saturated nitrogen-containing heteroaryl group;

[0053] Y is N or CH;

[0054] Each R 1 is cyano, hydroxy, hydroxyalkyl, halogen, haloalkyl, alkoxy, C-C1-C3 alkyl, -N(R 4 )2, -NR 4 C(O)C1-C3 alkyl or cycloalkyl;

[0055] Q is a bond, O or N;

[0056] Each R 2 is hydrogen or halogen;

[0057] Each R 3 is hydrogen, halogen or C1-C3 alkyl;

[0058] Each R 4 is hydrogen or C1-C3 alkyl; and

[0059] m is 0, 1, 2 or 3.

[0060] In one aspect of the invention, X is selected from indolizinyl, indolyl, isoindolyl, 3H-indolyl, indolyl, indazolyl, 1H-indazolyl, benzimidazolyl, benzothiazolyl, purinyl, 4H-quinolizinyl, quinolyl, isoquinolyl, cinnolinyl, phthaloyl, quinazolinyl, quinoxalinyl, 1,8-naphthyridinyl, pteridinyl, quinuclidine, carbazolyl, acridinyl, phenazinyl, phenthiazolyl, In one embodiment, X is R 1 -substituted indolin-1-yl.

[0061] In one embodiment, X is:

[0062]

[0063] where m, Y and R 1 As defined herein.

[0064] In one embodiment, m is 1 and R 1 is cyano, hydroxyl, hydroxyalkyl, halogen, haloalkyl, alkoxy, C-C1-C3 alkyl, -N(R 4 )2, -NR 4 C(O)C1-C3 alkyl or cycloalkyl.

[0065] In one embodiment, R 1 In one embodiment, R 1 In one embodiment, R 1 In one embodiment, halogen is chlorine or fluorine. In one embodiment, R 1 In one embodiment, the haloalkyl group is trifluoromethyl. In one embodiment, R 1 In one embodiment, Q is a bond, and C1-C3 alkyl is methyl, ethyl or isopropyl. In one embodiment, R 1 is Q-C1-C3 alkyl, wherein Q is O and C1-C3 alkyl is methyl, ethyl or isopropyl. 1 is Q-C1-C3 alkyl, wherein Q is N and C1-C3 alkyl is methyl, ethyl or isopropyl. 1 is hydroxyalkyl, wherein the hydroxyalkyl is hydroxymethyl. 1is an alkoxy group. In one embodiment, the alkoxy group is a methoxy group. In one embodiment, R 1 -N(R 4 )2. In one embodiment, each R 4 In one embodiment, each R 4 The group is a C1-C3 alkyl group. 1 For-NR 4 C(O)C1-C3 alkyl. In one embodiment, R 4 The group is hydrogen and the C1-C3 alkyl group is methyl. In one embodiment, R 1 is a cycloalkyl group, wherein the cycloalkyl group is a cyclopropyl group.

[0066] In one embodiment, m is 2, and each R 1 are independently cyano, hydroxy, hydroxyalkyl, halogen, haloalkyl, alkoxy, C1-C3 alkyl, -N(R 4 )2, -NR 4 C(O)C1-C3 alkyl or cycloalkyl. In certain embodiments, each R 1 In one embodiment, one R 1 is cyano, another R 1 is hydroxy, hydroxyalkyl, halogen, haloalkyl, alkoxy, C1-C3 alkyl, -N(R 4 )2, -NR 4 C(O)C1-C3 alkyl or cycloalkyl.

[0067] In one embodiment, X is:

[0068]

[0069] Each R 1 is hydrogen or methyl. In one embodiment, each R 1 It is methyl.

[0070] In one embodiment, R 2 In certain embodiments, the halogen is chlorine.

[0071] In one embodiment, R 3 is halogen or C1-C3 alkyl. In certain embodiments, halogen is fluorine. In other embodiments, R 3 It is a C1-C3 alkyl group, wherein the C1-C3 alkyl group is a methyl group.

[0072] In certain embodiments, the compound of Formula (Ia), Formula (Ib), Formula (IIa) or Formula (IIb) is:

[0073]

[0074]

[0075]

[0076] and pharmaceutically acceptable salts of the above compounds.

[0077] The compounds of Formula (Ia), Formula (Ib), Formula (IIa) or Formula (IIb) may be formulated into pharmaceutical compositions.

[0078] The compounds disclosed herein include all pharmaceutically acceptable isotopically labeled compounds, wherein one or more atoms of the compounds disclosed herein are replaced by an atom having the same atomic number and an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into the compounds disclosed herein include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, such as 2 H, 3 H, 11 C, 13 C, 14 C, 13 N, 15 N, 15 Oh, 17 Oh, 31 P, 32 P, 35 S, 18 F, 36 Cl, 123 I and 125 I. Such radiolabeled compounds can be used to help determine or measure the effectiveness of the compound, for example, by characterizing the binding affinity and kinetics of binding to ROCK and other modes of action. 3 H and carbon-14, 14 When labeled with C, certain compounds of the present disclosure can also be used to characterize the drug distribution and in vivo tissue expression of ROCK.

[0079] When using heavy isotopes such as deuterium, 2 When H-labeled, the compounds disclosed herein may exhibit therapeutic advantages, such as greater metabolic stability, leading to longer in vivo half-lives, which in turn may reduce therapeutic dosage requirements.

[0080] To better characterize substrate receptor occupancy or tissue expression in living samples statically or longitudinally, positron emitting isotopes (e.g. 11 C, 14 F, 15 O and 13N) substitution of atoms of the compounds disclosed herein may allow for positron emission tomography (PET) studies.

[0081] Isotopically-labeled compounds as disclosed herein can be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the relevant Examples and Schemes, by using an appropriate isotopically-labeled reagent in place of the non-labeled reagent previously employed.

[0082] Some compounds disclosed herein may exist in the form of stereoisomers. The compounds disclosed herein include all stereoisomers, either as pure individual stereoisomer preparations or as individually enriched preparations, and racemic mixtures of such stereoisomers as well as individual diastereomers and enantiomers that can be separated according to methods known to those skilled in the art. In addition, the compounds disclosed herein include all individual tautomeric states of the compounds and mixtures thereof.

[0083] Pharmaceutical composition

[0084] On the other hand, the present invention provides a pharmaceutical composition comprising a ROCK kinase inhibitor according to the present invention and a pharmaceutically acceptable carrier, excipient or diluent. The compounds of the present invention can be formulated by any method known in the art and can be prepared for administration by any route, including but not limited to parenteral, oral, sublingual, transdermal, topical, subcutaneous, intranasal, intratracheal or rectal administration. In certain embodiments, the compounds of the present invention are injected intravenously in a hospital setting. In certain other embodiments, administration may preferably be by oral route.

[0085] The characteristics of the carrier will depend on the route of administration. As used herein, the term "pharmaceutically acceptable" refers to a non-toxic substance that is compatible with biological systems such as cells, cell cultures, tissues or organisms, and does not interfere with the effectiveness of the biological activity of its active ingredients. Therefore, the composition of the present invention may also include diluents, fillers, salts, buffers, stabilizers, solubilizers and other materials well known in the art in addition to the inhibitor. The preparation of pharmaceutically acceptable preparations is described in, for example, Remington's Pharmaceutical Sciences, 18th edition, ed. A. Gennaro, Mack Publishing, Easton, Pennsylvania, 1990.

[0086] As used herein, the term pharmaceutically acceptable salt refers to a salt that retains the biological activity required by the above-mentioned compound and exhibits minimal or no undesirable toxicological effects. Examples of such salts include, but are not limited to, acid addition salts formed with inorganic acids (e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, etc.), and salts formed with organic acids such as acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic acid, benzoic acid, tannic acid, palmitic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, naphthalene disulfonic acid and polygalacturonic acid. The compounds can also be administered in the form of pharmaceutically acceptable quaternary salts known to those skilled in the art, specifically including quaternary ammonium salts of the formula -NR+Z-, wherein R is hydrogen, alkyl or benzyl, and Z is a counterion including chloride, bromide, iodide, -O-alkyl, tosylate, methanesulfonate, sulfonate, phosphate or carboxylate (e.g., benzoate, succinate, acetate, glycolate, maleate, malate, citrate, tartrate, ascorbate, benzoate, cinnamate, mandelate, benzyl ester and diphenyl acetate).

[0087] The active compound is contained in a pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver a therapeutically effective amount to the patient without causing severe toxic effects in the treated patient. For all of the above conditions, the dosage of the active compound is about 0.01 to 300 mg / kg per day, preferably 0.1 to 100 mg / kg per day, usually 0.5 to about 25 mg per kilogram of body weight per day of the subject. In a suitable carrier, a typical local dose range is 0.01-3% wt / wt. The effective dose range of the pharmaceutically acceptable derivative can be calculated based on the weight of the parent compound to be delivered. If the derivative itself shows activity, the weight of the derivative can be used as described above or the effective dose can be estimated by other means known to those skilled in the art.

[0088] Pharmaceutical compositions comprising compounds of the invention are useful in the methods described herein.

[0089] How to use

[0090] In another aspect, the present invention provides a method for inhibiting ROCK activity in cells, comprising contacting cells in need of inhibiting ROCK activity with a therapeutically effective amount of a compound of formula (Ia), (Ib), (IIa) or (IIb), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the compound or a pharmaceutically acceptable salt thereof.

[0091] It is believed that the compositions and methods provided herein are particularly useful for inhibiting ROCK activity in cells. In one embodiment, cells that need to inhibit ROCK activity are contacted with an effective amount of a compound of formula (Ia), (Ib), (IIa) or (IIb) to negatively regulate the activity of ROCK kinase. In other embodiments, a therapeutically effective amount of a pharmaceutically acceptable salt or pharmaceutical composition containing a compound of formula (Ia), (Ib), (IIa) or (IIb) may be used. In certain embodiments, contacting cells with an effective amount or a therapeutically effective amount of a compound of formula (Ia), (Ib), (IIa) or (IIb) occurs in vivo. In certain embodiments, contacting cells with an effective amount or a therapeutically effective amount of a compound of formula (Ia), (Ib), (IIa) or (IIb) occurs in vitro. In one embodiment, the therapeutically effective amount of a compound of formula (Ia), (Ib), (IIa) or (IIb) is about 0.01 mg / kg to 300 mg / kg per day. In one embodiment, the therapeutically effective amount of the compound of Formula (Ia), Formula (Ib), Formula (IIa) or Formula (IIb) is about 0.1 mg / kg to 100 mg / kg per day.

[0092] In one embodiment, the compound of formula (Ia), formula (Ib), formula (IIa) or formula (IIb) preferentially inhibits the activity of ROCK2 kinase compared to ROCK1 kinase. In another embodiment, the compound of formula (Ia), formula (Ib), formula (IIa) or formula (IIb) inhibits the activity of ROCK2 kinase and ROCK1 kinase to a similar extent. In one embodiment, the therapeutically effective amount of the compound of formula (Ia), formula (Ib), formula (IIa) or formula (IIb) is about 0.01 mg / kg to 300 mg / kg per day. In one embodiment, the therapeutically effective amount of the compound of formula (Ia), formula (Ib), formula (IIa) or formula (IIb) is about 0.1 mg / kg to 100 mg / kg per day.

[0093] In one embodiment, a method for treating a patient suffering from cerebral cavernous malformation syndrome (CCM) is provided, comprising administering to the patient a therapeutically effective amount of a compound of Formula (Ia), Formula (Ib), Formula (IIa) or Formula (IIb), or a pharmaceutically acceptable salt thereof, alone or in combination with a pharmaceutically acceptable carrier, excipient or diluent.

[0094] In another embodiment, a method for treating a cardiovascular disease associated with increased vascular tone is provided, comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula (Ia), formula (Ib), formula (IIa) or formula (IIb) or a pharmaceutical composition thereof. In one embodiment, the cardiovascular disease is hypertension, atherosclerosis, ischemic stroke, coronary artery spasm, cerebral vasospasm, angina pectoris and erectile dysfunction.

[0095] In an embodiment, a method for treating a disease involving increased non-vascular smooth muscle contractility is provided, comprising administering a therapeutically effective amount of a compound of formula (Ia), formula (Ib), formula (IIa) or formula (IIb) or a pharmaceutical composition thereof to a patient in need thereof. In one embodiment, the disease involves asthma and glaucoma involving increased non-vascular smooth muscle contractility.

[0096] By negatively regulating the activity of ROCK, particularly in the case of cells overexpressing the ROCK enzyme or autosomal dominant mutations that activate the ROCK enzyme, in certain embodiments, the method is designed to modulate the activity of ROCK to inhibit the formation of new CCMs and / or reduce or eradicate existing CCMs in treated patients, or to treat certain cardiovascular diseases or diseases involving elevated contractility of non-vascular smooth muscle. Cells / patients may be contacted with a single dose or multiple doses according to a specific treatment regimen to achieve the desired negative regulation of ROCK. For example, well-known methods including CT scans or MRI scans may be used to monitor the size and number of CCMs to assess the effectiveness of the treatment, and the attending physician may adjust the dose accordingly.

[0097] The concentration and route of administration to the patient may vary depending on the severity of the disease. The compound, its pharmaceutically acceptable salt, and pharmaceutical compositions comprising such compound and salt may also be co-administered with other compounds or used in combination with other treatment methods (e.g., surgical intervention) as a preoperative or postoperative adjuvant. Well-known assays including those in Example A may be used to monitor the extent of ROCK kinase inhibition in patients to obtain the effectiveness of treatment, as well as other cranial imaging techniques and instruments, such as CT scans, MRI, X-rays, etc., and the attending physician may adjust the dosage accordingly.

[0098] Reaction Schemes and Examples

[0099] The compounds of the present invention can be prepared using commercially available reagents, using the synthetic methods and reaction schemes described herein, or using other reagents and conventional methods well known to those skilled in the art.

[0100] For example, compounds of the present invention can be prepared according to general reaction schemes I and II.

[0101] Solution 1

[0102]

[0103] Scheme I illustrates the preparation of compounds of formula (I) of the present invention. 2 Substituted haloisoquinoline A undergoes an aromatic electrophilic substitution reaction with sulfonyl chloride acid at elevated temperature to give halosulfonated intermediate B (step 1). Halosulfonated intermediate B reacts under acidic conditions, such as HCl, at elevated temperature to give sulfonic acid substituted isoquinolinone C (step 2). Sulfonic acid is usually converted to sulfonyl chloride using sulfonyl chloride in DMF at reduced temperature (step 3). 1 Addition of the substituted nitrogen-containing heteroaryl group X is typically performed under basic conditions using a suitable base such as TEA in a solvent such as DCM to afford compounds E of formula (I) (step 4).

[0104]

[0105] Scheme II illustrates the preparation of compounds of formula (II) of the present invention, wherein R 3 In the presence of sulfuric acid, the temperature is lowered (eg, 0° C.), and nitric acid is used to nitrate the halogen-substituted isoquinoline A to obtain a nitro-substituted compound B. Boric acid R 3 B(OH)2 is coupled with intermediate B via Suzuki reaction using a suitable palladium catalyst (e.g. Pd(dddf)Cl2) in the presence of a suitable base (e.g. CsCO3 in dioxane) to generate R 3 -substituted compound C. Nitro-containing compound C is reduced to amine D. Amine D reacts with sodium nitrite under acidic conditions and then treated with copper chloride and SO2 to obtain sulfonyl chloride E. The added R 1 -substituted nitrogen-containing heteroaryl X is typically treated under basic conditions using an appropriate base such as TEA in a solvent such as DCM to give compound F. Reaction with m-CPBA gives N-oxide G. Treatment of N-oxide G with acetic anhydride under heating gives acetate which undergoes spontaneous 2,3-σ rearrangement. Deprotection of the acetate gives 1-oxo compound H.

[0106] The following examples are intended to illustrate certain other embodiments of the present invention, but are not intended to limit the scope of the present invention.

[0107] Example 1

[0108]

[0109] 5-((4-Methylindolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

[0110]

[0111] Compound 1-2: Under N2 atmosphere, 1-chloroisoquinoline (5g, 30.5mmol) was stirred in a solution of sulfonyl chloride (50mL, 750.9mmol) at 150°C for 12h. The reaction mixture was quenched by adding ice, then diluted with H2O (300mL), and extracted with DCM (100mL×3). The combined organic layer was washed with saturated brine (300mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 0: 1 to 10: 1). Compound 1-2 was obtained as a yellow solid, which was 1-chloroisoquinoline-5-sulfonyl chloride (19.1g, 72.9mmol, 47.7% yield).

[0112] Compound 1-3: Under N2 atmosphere, a solution of 1-chloroisoquinoline-5-sulfonyl chloride (2.5 g, 9.54 mmol) in aqueous HCl (30 mL, 9N, 276.9 mmol) was stirred at 100 ° C for 12 h. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The crude product compound 1-3 was obtained in the form of a yellow solid, which was 1-hydroxyisoquinoline-5-sulfonic acid (2.8 g, crude product), which was used in the next step without further purification.

[0113] Compound 1-4: DMF (273 μL, 3.55 mmol) was added to a solution of 1-hydroxyisoquinoline-5-sulfonic acid (0.8 g, 3.55 mmol) in SOCl2 (40 mL) at 0°C, and the mixture was stirred at 15°C for 30 minutes under N2 atmosphere. The mixture was then stirred at 35°C for 2.5 h under N2 atmosphere. The reaction mixture was diluted with H2O (100 mL) and extracted with DCM (200 mL×3). The combined organic layers were washed with saturated brine (100 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The crude product compound 1-4 was obtained as a yellow solid, which was 1-hydroxyisoquinoline-5-sulfonyl chloride (0.3 g, 1.23 mmol, 34.6% yield). The compound was used in the next step without further purification.

[0114] Example 1: TEA (128.5 μL, 923.4 μmol) and 4-methylindoline (90.1 mg, 677.1 μmol) were added to a solution of 1-hydroxyisoquinolyl-5-sulfonyl chloride (150 mg, 615.6 μmol) in DCM (10 mL). The mixture was stirred at 15 ° C for 5 h under N2 atmosphere. The reaction mixture was diluted with H2O (60 mL) and extracted with DCM (200 mL×2). The combined organic layer was washed with saturated brine (80 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 1: 1).

[0115] Example 1 was obtained as a white solid as 5-(4-methylindol-1-yl)sulfonyl-2H-isoquinolin-1-one (90 mg, 252 μmol, 41.1% yield). LC-MS: [M+1] 340.09.

[0116] 1 H NMR (400MHz, DMSO-d6): δ11.65(br,1H),8.48(d,J=8Hz,1H),8.26(d,J=7.6Hz,HI),7.59(t,J=7.6Hz,1H),7.34( t,J=6Hz,1H),7.20~7.02(m,3H),6.81(d,J=7.6Hz,1H),4.04(t,J=8.0Hz,2H),2.89(t,J=8Hz,2H),2.11(s,3H).

[0117] Example 2

[0118]

[0119] 5-((5-Hydroxyindolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

[0120]

[0121] Compound 2-2: TEA (318 μL, 2.29 mmol) and indolin-5-ol (226.9 mg, 1.68 mmol) were added to a solution of 1-chloroisoquinoline-5-sulfonyl chloride (0.4 g, 1.53 mmol) in DCM (10 mL). The mixture was stirred at 25 ° C for 12 h under N2 atmosphere. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 1: 1). Compound 2-2 was obtained as a yellow solid, which was 1-[(1-chloro-5-isoquinolyl)sulfonyl]indolin-5-ol (350 mg, 970.0 μmol, 63.6% yield).

[0122] Example 2: A solution of 1-[(1-chloro-5-isoquinolyl)sulfonyl]indolin-5-ol (330 mg, 914.6 μmol) in HCl (10 mL, 9N) was stirred at 90 ° C under N2 atmosphere for 12 h. The reaction mixture was quenched by adding saturated NaHCO3 solution until pH = 7, then diluted with H2O (30 mL) and extracted with EtOAc (10 mL×3). The combined organic layer was washed with saturated brine (30 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Waters Xbridge Prep O BD C18 150 mm x 40 mm x 10 μm; mobile phase: [A, water (10 mM NH4HCO3)-B, ACN]; B%: 25%-55%, 11 minutes). Example 2 was obtained as a yellow solid as 5-(5-hydroxyindolin-1-yl)sulfonyl-2H-isoquinolin-1-one (6 mg, 17.1 μmol, 1.87% yield). LC-MS: [M+1] 342.07.

[0123] 1 H NMR (400MHz, CDCl3): δ10.16 (br, 1H), 8.73 (d, J = 8 Hz, 1H), 8.20 (d, J = 8.8 Hz, 1H), 7.56-7.44 (m, 2H), 7.38-7.30 (m, 1H), 6.74 (s, 1H), 6.43 (d, J = 6.4 Hz, 1H), 6.35 (d, J = 8.4 Hz, 1H), 3.54 (t, J = 8.4 Hz, 2H), 2.93 (d, J = 8.4 Hz, 2H). One proton was exchanged.

[0124] Example 3

[0125]

[0126] 5-(Indolin-1-ylsulfonyl)isoquinolin-1(2H)-one

[0127]

[0128] Compound 3-1: TEA (238.9 μL, 1.72 mmol) and indoline (109.1 mg, 915.6 μmol) were added to a solution of 1-chloroisoquinoline-5-sulfonyl chloride (0.3 g, 1.14 mmol) in DCM (8 mL). The mixture was stirred at 15 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (60 mL) and extracted with EtOAc (100 mL×3). The combined organic layer was washed with saturated brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The crude product compound 3-1 was obtained as a yellow solid, which was 1-chloro-5-indoline-1-ylsulfonyl-isoquinoline (0.2 g, crude product). Compound 3-1 can be used in the next step without further purification.

[0129] Example 3: A mixture of 1-chloro-5-indoline-1-ylsulfonyl-isoquinoline (0.2 g, 580.02 μmol, 1 eq) in aqueous HCl (5 mL, 12 M) was stirred at 100 ° C for 12 h under N2 atmosphere. The reaction mixture was quenched by adding ice water (15 mL) at 0 ° C, and then NaHCO3 was added to the mixture to adjust to pH 8 and filtered. The filtrate was concentrated under reduced pressure to obtain a residue. The residue was distributed between H2O (60 mL) and DCM (100 mL×3). The combined organic layer was washed with saturated brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 0: 1). Example 3 was obtained as a white solid as 5-indolyl-1-ylsulfonyl-2H-isoquinolin-1-one (40 mg, 117.5 μmol, 20.2% yield). LC-MS: [M+1] 326.07.

[0130] 1 H NMR (400 MHz, MeOD): δ 8.58 (d, J = 8.4 Hz, 1H), 8.34 (d, J = 8 Hz, 1H), 7.61 (t, J = 8 Hz, 1H), 7.47 (d, J = 8 Hz, 1H), 7.38 (d, J = 7.6 Hz, 1H), 7.25-7.10 (m, 3H), 7.00 (t, J = 8 Hz, 1H), 4.03 (t, J = 8 Hz, 2H), 2.85 (t, J = 8.4 Hz, 2H). One proton was exchanged.

[0131] Example 4

[0132]

[0133] 5-((6-Chloroindol-1-yl)sulfonyl)isoquinolin-1(2H)-one

[0134]

[0135] Example 4: TEA (85.7 μL, 615.6 μmol) and 6-chloroindoline (56.7 mg, 369.4 μmol) were added to a solution of 1-hydroxyisoquinoline-5-sulfonyl chloride (100.0 mg, 410.4 μmol) in DCM (5 mL). The mixture was stirred at 15 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (50 mL) and extracted with DCM (150 mL×2). The combined organic layer was washed with saturated brine (100 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, ether: ethyl acetate = 8: 1 to 0: 1). 40 mg of Example 4 was obtained as a white solid, which is 5-(6-chloroindoline-1-yl)sulfonyl-2H-isoquinolin-1-one. LC-MS: [M+1] 360.03.

[0136] 1 H NMR (400 MHz, MeOD): δ 8.60 (d, J = 8 Hz, 1H), 8.35 (d, J = 8 Hz, 1H), 7.63 (t, J = 8 Hz, 1H), 7.45 (d, J = 2 Hz, 1H), 7.37 (d, J = 7.6 Hz, 1H), 7.26 (d, J = 7.6 Hz, 1H), 7.10 (d, J = 7.6 Hz, 1H), 7.02-6.96 (m, 1H), 4.03 (t, J = 8.4 Hz, 2H), 2.87 (t, J = 8.4 Hz, 2H). One proton was exchanged.

[0137] Example 5

[0138]

[0139] 5-((6-Fluoroindol-1-yl)sulfonyl)isoquinolin-1(2H)-one

[0140]

[0141] Example 5: 6-fluoroindoline (75.9 mg, 554.0 μmol) and TEA (128.5 μL, 923.3 μmol) were added to a solution of 1-hydroxyisoquinoline-5-sulfonyl chloride (150.0 mg, 615.6 μmol) in DCM (5 mL). The mixture was stirred at 15 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (30 mL) and extracted with DCM (100 mL×2). The combined organic layer was washed with saturated brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 0: 1). Example 5 was obtained as a yellow solid, which was 5-(6-fluoroindoline-1-yl)sulfonyl-2H-isoquinolin-1-one (80 mg, 204.3 μmol, 33.2% yield). LC-MS: [M+1] 344.06.

[0142] 1 H NMR (400 MHz, MeOD): δ8.59 (d, J = 8.0 Hz, 1H), 8.36 (d, J = 7.6 Hz, 1H), 7.62 (t, J = 8 Hz, 1H), 7.38 (d, J = 8 Hz, 1H), 7.26 (d, J = 7.6 Hz, 1H), 7.22-7.17 (m, 1H), 7.12-7.06 (m, 1H), 6.75-6.66 (m, 1H), 4.03 (t, J = 8.4 Hz, 2H), 2.87 (t, J = 8.4 Hz, 2H). One proton was exchanged.

[0143] Example 6

[0144]

[0145] 5-((6-Methylindolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

[0146]

[0147] Compound 6-1: TEA (239 μL, 1.72 mmol) and 6-methylindoline (121 mg, 916 μmol) were added to a solution of 1-chloroisoquinoline-5-sulfonyl chloride (0.3 g, 1.14 mmol) in DCM (8 mL). The mixture was stirred at 15 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (60 mL) and extracted with EtOAc (100 mL×3). The combined organic layer was washed with saturated brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The crude product compound 6-1 was obtained as a yellow solid, which was 1-chloro-5-(6-methylindoline-1-yl)sulfonyl-isoquinoline (0.2 g, crude product). The crude product can be used in the next step without further purification.

[0148] Example 6: A mixture of 1-chloro-5-(6-methylindolin-1-yl)sulfonyl-isoquinoline (0.2 g, 557 μmol) in aqueous HCl (5 mL, 12 M) was degassed and purged 3 times with N2, and then the mixture was stirred at 100 ° C for 12 h under N2 atmosphere. The reaction mixture was quenched by adding ice water (15 mL) at 0 ° C, and then NaHCO3 was added to the mixture to adjust to pH = 8 and filtered. The filtrate was concentrated under reduced pressure to obtain a residue. The reaction mixture was diluted with H2O (60 mL) and extracted with DCM (100 mL×3). The combined organic layer was washed with saturated brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 0: 1). Example 6 was obtained as 5-(6-methylindolin-1-yl)sulfonyl-2H-isoquinolin-1-one (40 mg, 107.4 μmol, 19.2% yield) as a white solid. LC-MS: [M+1] 340.09.

[0149] 1 H NMR (400 MHz, MeOD): δ 8.57 (d, J = 8.4 Hz, 1H), 8.32 (d, J = 7.6 Hz, 1H), 7.59 (t, J = 8 Hz, 1H), 7.38 (d, J = 8 Hz, 1H), 7.30 (s, 1H), 7.22 (d, J = 8 Hz, 1H), 6.99 (d, J = 7.6 Hz, 1H), 6.82 (d, J = 7.6 Hz, 1H), 4.01 (t, J = 8 Hz 2H), 2.79 (t, J = 8 Hz 2H), 2.31 (s, 3H). One proton was exchanged.

[0150] Example 7

[0151]

[0152] 5-((4-(Hydroxymethyl)indolin-1-ylsulfonyl)isoquinolin-1(2H)-one

[0153]

[0154] Example 7: TEA (257.0 μL, 1.85 mmol) and indolin-4-ylmethanol (202.0 mg, 1.35 mmol) were added to a solution of 1-oxo-2H-isoquinoline-5-sulfonyl chloride (0.3 g, 1.23 mmol) in DCM (10 mL). The mixture was stirred at 25 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (15 mL) and extracted with DCM (5 mL×3). The combined organic layer was washed with saturated brine (15 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 1: 0 to 0: 1). Example 7 was obtained as a dark brown solid, 5-[4-(hydroxymethyl)indolin-1-yl]sulfonyl-2H-isoquinolin-1-one (35 mg, 91.59 μmol, 7.44% yield). LC-MS: [M+1] 356.08.

[0155] 1 H NMR (400 MHz, MeOD): δ 8.56 (d, J = 8.0 Hz, 1H), 8.34 (d, J = 7.6 Hz, 1H), 7.58 (t, J = 7.6 Hz, 1H), 7.50 (d, J = 8 Hz, 2H), 7.22 (d, J = 8 Hz, 1H), 7.17 (t, J = 8 Hz, HI), 7.04 (d, J = 7.6 Hz, 1H), 4.46 (s, 2H), 4.06 (t, J = 8.4 Hz, 2H), 2.91 (t, J = 8.4 Hz, 2H). Two protons were exchanged.

[0156] Example 8

[0157]

[0158] 5-((4-(Trifluoromethyl)indolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

[0159]

[0160] Example 8: TEA (128.5 μL, 923.3 μmol) and 4-(trifluoromethyl)indoline (103.6 mg, 554.0 μmol) were added to a solution of 1-hydroxyisoquinoline-5-sulfonyl chloride (150 mg, 615 μmol) in DCM (5 mL). The mixture was stirred at 15 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (30 mL) and extracted with DCM (75 mL×2). The combined organic layer was washed with saturated brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 0: 1).

[0161] Example 8 was obtained as a white solid, 5-[4-(trifluoromethyl)indolin-1-yl]sulfonyl-2H-isoquinolin-1-one (20 mg, 44.2 μmol, 7.18% yield). LC-MS: [M+1] 394.06.

[0162] 1 H NMR (400 MHz, MeOD): δ 8.60 (d, J = 8 Hz, 1H), 8.38 (d, J = 7.6 Hz, 1H), 7.70 (d, J = 8.4 Hz, 1H), 7.63 (t, J = 8 Hz, 1H), 7.40-7.20 (m, 4H), 4.10 (t, J = 8.4 Hz, 2H), 3.05 (t, J = 8.4 Hz, 2H). One proton was exchanged.

[0163] Example 9

[0164]

[0165] 5-((4-cyclopropylindolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

[0166]

[0167] Example 9: TEA (128.5 μL, 923.3 μmol) and 4-cyclopropylindoline (88.2 mg, 554.0 μmol) were added to a solution of 1-hydroxyisoquinoline-5-sulfonyl chloride (150 mg, 615.5 μmol) in DCM (5 mL). The mixture was stirred at 15 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (40 mL) and extracted with DCM (100 mL×2). The combined organic layer was washed with saturated brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 0: 1). Example 9 was obtained as a white solid, 5-(4-cyclopropylindolin-1-yl)sulfonyl-2H-isoquinolin-1-one (25 mg, 57.8 μmol, 9.39% yield). LC-MS: [M+1] 366.10.

[0168] 1 H NMR (400MHz, MeOD): δ8.57(d,J=7.6Hz,1H),8.33(d,J=7.6Hz,1H),7.59(t,J=8Hz,1H),7.33(d,J=8Hz,1H),7.26(d,J=8Hz,1H),7.19(d,J=8Hz,1H), 7.07(t,J=7.6Hz,1H),6.56(d,J=7.6Hz,1H),4.05(t,J=8.4Hz,1H),2.93( t,J=8.4Hz,2H),1.73~1.67(m,1H),0.88~0.86(m,2H),0.57~0.55(m,2H).

[0169] Example 10

[0170]

[0171] 5-((4-ethylindolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

[0172]

[0173] Compound 10-1: TEA (128.5 μL, 923.3 μmol) and 4-vinyl indoline (98.3 mg, 677.1 μmol) were added to a solution of 1-hydroxyisoquinoline-5-sulfonyl chloride (150 mg, 615.5 μmol) in DCM (5 mL). The mixture was stirred at 15 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (20 mL) and extracted with DCM (80 mL×2). The combined organic layer was washed with saturated brine (30 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 0: 1). Compound 10-1, 5-(4-vinylindolin-1-yl)sulfonyl-2H-isoquinolin-1-one, was obtained as a yellow solid (50 mg, 141.8 μmol, 23.05% yield).

[0174] Example 10: A mixture of 5-(4-vinylindolyl-1-yl)sulfonyl-2H-isoquinolin-1-one (50 mg, 141.8 μmol), Pd / C (5 mg, 10 wt%) in methanol (5 mL) was degassed and purged with H2 three times, and then the mixture was stirred at 15°C for 12 h under a H2 atmosphere (15 psi). The reaction mixture was filtered and the filter was concentrated. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 5: 1 to 0: 1). The residue was purified by preparative HPLC (chromatographic column: Waters Xbridge 150 mm x 50 mm x 10 μm; mobile phase: [A, water (10 mM NH4HCO3)-B, ACN]; B%: 30%-60%, 12 minutes). Example 10 was obtained as 5-(4-ethylindolin-1-yl)sulfonyl-2H-isoquinolin-1-one (14 mg, 39.4 μmol, 27.7% yield) as a white solid. LC-MS: [M+1] 354.10.

[0175] 1 H NMR (400 MHz, MeOD): δ8.58 (d, J = 7.6 Hz, 1H), 8.34 (d, J = 7.6 Hz, 1H), 7.60 (t, J = 8 Hz, 1H), 7.32 (t, J = 7.2 Hz, 2H), 7.18 (d, J = 4.0 Hz, 1H), 7.13 (t, J = 7.6 Hz, 1H), 6.88 (d, J = 4 Hz, 1H), 4.03 (t, J = 8.4 Hz, 2H), 2.76 (t, J = 8.4 Hz, 2H), 2.50-2.44 (m, 2H), 1.06 (t, J = 7.6 Hz, 3H). One proton was exchanged.

[0176] Embodiment 11

[0177]

[0178] 5-((4-(1-Hydroxyethyl)indolin-1-ylsulfonyl)isoquinolin-1(2H)-one

[0179]

[0180] Example 11: TEA (209.3 μL, 1.50 mmol) and 1-indolin-4-ylethanol (180 mg, 1.10 mmol) were added to a solution of 1-oxo-2H-isoquinoline-5-sulfonyl chloride (244.2 mg, 1.00 mmol) in DCM (5 mL). The mixture was stirred at 25 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (60 mL) and extracted with DCM (150 mL×2). The combined organic layer was washed with saturated brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 0: 1). The residue was purified by preparative HPLC (column: Waters Xbridge 150 mm x 25 mm x 5 μm; mobile phase: [A, water (10 mM NH4HCO3)-B, ACN]; B%: 10%-40%, 10 min). Example 11 was obtained as a white solid as 5-[4-(1-hydroxyethyl)indolin-1-yl]sulfonyl-2H-isoquinolin-1-one (40 mg). LC-MS: [M+1] 370.10.

[0181] 1 H NMR (400 MHz, MeOD): δ 8.57 (d, J = 7.6 Hz, 1H), 8.35 (d, J = 8 Hz, 1H), 7.59 (t, J = 8 Hz, 1H), 7.38 (d, J = 8, 1H), 7.34 (d, J = 8 Hz, 1H), 7.21-7.15 (m, 2H), 7.11 (d, J = 7.6 Hz, 1H), 4.88-4.71 (m, 1H), 4.07-4.01 (m, 2H), 2.89-2.80 (m, 2H), 1.26 (d, J = 6.4 Hz, 3H). Two protons were exchanged.

[0182] Example 12

[0183]

[0184] 5-((3,3-Dimethylindolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

[0185]

[0186] Compound 12-1: TEA (238.9 μL, 1.72 mmol) and 3,3-dimethylindoline (134.8 mg, 915.6 μmol) were added to a solution of 1-chloroisoquinoline-5-sulfonyl chloride (0.3 g, 1.14 mmol) in DCM (8 mL). The mixture was stirred at 15 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (60 mL) and extracted with EtOAc (100 mL×3). The combined organic layer was washed with saturated brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The crude product compound 12-1 was obtained as a yellow solid, which was 1-chloro-5-(3,3-dimethylindoline-1-yl)sulfonyl-isoquinoline (0.2 g, crude product). The crude product can be used in the next step without further purification.

[0187] Example 12: A mixture of 1-chloro-5-(3,3-dimethylindolin-1-yl)sulfonylisoquinoline (0.2 g, 536.3 μmol) in aqueous HCl (5 mL, 12 M) was degassed and purged with N2 three times, and then the mixture was stirred at 100 ° C for 12 h under N2 atmosphere. The reaction mixture was quenched by adding ice water (15 mL) at 0 ° C, and then NaHCO3 was added to the mixture to adjust to pH 8, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was diluted with H2O (60 mL) and extracted with DCM (100 mL×3). The combined organic layer was washed with saturated brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 0: 1). Example 12 was obtained as a white solid, 5-(3,3-dimethylindolin-1-yl)sulfonyl-2H-isoquinolin-1-one (12 mg, 31.5 μmol, 5.88% yield). LC-MS: [M+1] 354.10.

[0188] 1 H NMR (400 MHz, MeOD): δ 8.58, (d, J = 8 Hz, 1H), 8.42 (d, J = 8 Hz, 1H), 7.62 (t, J = 7.6 Hz, 1H), 7.48 (d, J = 8 Hz, 1H), 7.43 (d, J = 7.6 Hz, 1H), 7.23 (d, J = 7.6 Hz, 1H), 7.18 (t, J = 7.6 Hz, 1H), 7.11 (d, J = 7.6 Hz, 1H), 7.02 (t, J = 7.6 Hz, 1H), 3.73 (s, 2H), 1.08 (s, 6H). One proton was exchanged.

[0189] Embodiment 13

[0190]

[0191] 5-((3-Methylindolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

[0192]

[0193] Example 13: TEA (85.6 μL, 615.6 μmol) and 3-methylindoline (55.7 mg, 328.3 μmol) were added to a solution of 1-hydroxyisoquinoline-5-sulfonyl chloride (100 mg, 410.4 μmol) in DCM (5 mL). The mixture was stirred at 15 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (50 mL) and extracted with DCM (150 mL×2). The combined organic layer was washed with saturated brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 0: 1). 70 mg of Example 13 was obtained as a white solid, which is 5-(3-methylindoline-1-yl)sulfonyl-2H-isoquinolin-1-one. LC-MS: [M+1] 340.09.

[0194] 1 H NMR (400MHz, DMSO-d6): δ11.67(s,1H),8.49(d,J=8Hz,1H),8.30(d,J=8Hz,1H),7.61(t,J=7.6Hz,1H),7.31~7.30(m,2H),7 .20~7.13(m,3H),7.01(t,J=7.6Hz,1H),4.19(t,J=9.2Hz,1H),3.57-3.53(m,1H),3.29~3.24(m,1H),1.05(d,J=6.8Hz,3H).

[0195] Embodiment 14

[0196]

[0197] 5-((4-isopropylindolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

[0198]

[0199] Compound 14-1: TEA (71.5 μL, 513.8 μmol) and 4-isopropenyl indoline (60 mg, 376.82 μmol) were added to a solution of 1-hydroxyisoquinoline-5-sulfonyl chloride (83.4 mg, 342.6 μmol) in DCM (5 mL). The mixture was stirred at 15 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (10 mL) and extracted with DCM (50 mL×3). The combined organic layer was washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 1: 1). Compound 14-1, 5-(4-isopropenylindolin-1-yl)sulfonyl-2H-isoquinolin-1-one, was obtained as a yellow solid (50 mg, 136.4 μmol, 39.8% yield).

[0200] Example 14: To a solution of 5-(4-isopropenylindoline-1-yl)sulfonyl-2H-isoquinolin-1-one (50 mg, 136.45 μmol, 1 eq) in methanol (5 mL) was added Pd / C (5 mg, 10% purity) under H2. The mixture was stirred at 15°C under a pressure of 15 psi for 12 h. The reaction mixture was filtered and the filtrate was concentrated. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 5: 1 to 0: 1). The residue was purified by preparative HPLC (chromatographic column: Waters Xbridge 150 mm x 50 mm x 10 um; mobile phase: [A, water (10 mM NH4HCO3)-B, ACN]; B%: 30%-60%, 12 minutes). Example 14 was obtained as 5-(4-isopropylindolin-1-yl)sulfonyl-2H-isoquinolin-1-one (8 mg, 21.50 μmol, 15.7% yield) as a white solid. LC-MS: [M+1] 368.12.

[0201] 1 H NMR (400 MHz, MeOD): δ8.57 (d, J = 8.0 Hz, 1H), 8.34 (d, J = 6.8 Hz, 1H), 7.59 (t, J = 8 Hz, 1H), 7.32-7.26 (m, 2H), 7.15 (t, J = 7.6 Hz, 2H), 6.95 (d, J = 7.6 Hz, 1H), 4.00 (t, J = 8 Hz, 2H), 2.81-2.71 (m, 3H), 1.10 (d, J = 6.8 Hz, 6H). One proton was exchanged.

[0202] Embodiment 15

[0203]

[0204] 1-((1-Oxy-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

[0205]

[0206] Example 15: TEA (173.8 μL, 1.25 mmol) and indoline-6-carbonitrile (120 mg, 832.3 μmol) were added to a solution of 1-hydroxyisoquinoline-5-sulfonyl chloride (202.8 mg, 832.3 μmol) in DCM (5 mL). The mixture was stirred at 25 ° C for 5 h under N2 atmosphere. The reaction mixture was quenched by adding H2O (40 mL), and then the reaction mixture was extracted with DCM (100 mL×2). The combined organic layer was washed with saturated brine 50 (mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 0: 1). Example 15 was obtained as 1-[(1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrile (80 mg, 215.8 μmol, 25.9% yield) as a yellow solid. LC-MS: [M+1] 351.07.

[0207] 1 H NMR (400MHz, DMSO-d6): δ11.68(s,1H),8.51(d,J=7.6Hz,1H),8.37(d,J=6.4Hz,1H),7.64(t,J=8Hz,1 H), 7.58 (s, 1H), 7.47-7.40 (m, 3H), 7.11 (d, J = 7.2Hz, 1H), 4.06 (t, J = 8Hz, 2H), 3.07 (t, J = 8.0Hz, 2H).

[0208] Example 16

[0209]

[0210] 5-((4,6-Dichloroindolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

[0211]

[0212] Example 16: TEA (151.3 μL, 1.09 mmol) and 4,6-dichloroindoline (150 mg, 797.6 μmol) were added to a solution of 1-oxo-2H-isoquinoline-5-sulfonyl chloride (176.6 mg, 725.1 μmol) in DCM (3 mL). The mixture was stirred at 25 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (60 mL) and extracted with DCM (150 mL×2). The combined organic layer was washed with saturated brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 0: 1). The residue was purified by preparative HPLC (column: Waters Xbridge Prep OBD C18 150 mm x 40 mm x 10 μm; mobile phase: [A, water (10 mM NH4HCO3)-B, ACN]; B%: 35%-55%, 11 minutes). Example 16 was obtained as a white solid as 5-(4,6-dichloroindolin-1-yl)sulfonylisoquinolin-1-ol (60 mg, 150.5 μmol, 20.7% yield). LC-MS: [M+1] 393.99.

[0213] 1 H NMR (400MHz, DMSO-d6): δ8.53(d,J=7.6Hz,1H),8.31(d,J=8Hz,2H),7.67(t,J=8Hz,1H),7.39(d,J=7 .6Hz, 1H) 7.23 (d, J = 8.8Hz, 2H), 7.10 (d, J = 7.6Hz, 1H), 4.09 (t, J = 8.4Hz, 2H), 3.03 (t, J = 8.4Hz, 2H).

[0214] Embodiment 17

[0215]

[0216] 4-Cyclopropyl-1-((1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

[0217]

[0218] Example 17: TEA (85.6 μL, 615.6 μmol) and 4-cyclopropylindoline-6-carbonitrile (37.8 mg, 205.2 μmol) were added to a solution of 1-oxo-2H-isoquinoline-5-sulfonyl chloride (50 mg, 205.2 μmol) in DCM (5 mL). The mixture was stirred at 25 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (15 mL) and extracted with DCM (80 mL×2). The combined organic layer was washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (chromatographic column: Xtimate C18 150 mm x 25 mm x 5 μm; mobile phase: [A, water (10 mM NH4HCO3)-B, ACN]; B%: 50%-80%, 10 minutes). Example 17 was obtained as 4-cyclopropyl-1-[(1-hydroxy-5-isoquinolinyl)sulfonyl]indoline-6-carbonitrile as a yellow solid (19 mg, 47.5 μmol, 23.1% yield). LC-MS: [M+1] 391.10.

[0219] 1 H NMR (400 MHz, MeOD): δ8.62 (d, J = 8 Hz, 1H), 8.39 (d, J = 6.8 Hz, 1H), 7.65 (t, J = 7.8 Hz, 1H), 7.54 (s, 1H), 7.35 (d, J = 7.6 Hz, 1H), 7.27 (d, J = 7.6 Hz, 1H), 6.92 (s, 1H), 4.09 (t, J = 8.4 Hz, 2H), 3.07 (t, J = 8.4 Hz, 2H), 1.79-1.77 (m, 1H), 0.98-0.95 (m, 2H), 0.68-0.66 (m, 2H). One proton was exchanged.

[0220] Embodiment 18

[0221]

[0222] 4-Methyl-1-((1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

[0223]

[0224] Example 18: TEA (65.9 μL, 474.0 μmol) and 4-methylindoline-6-carbonitrile (50 mg, 316.0 μmol) were added to a solution of 1-oxo-2H-isoquinoline-5-sulfonyl chloride (100 mg, 410.4 μmol) in DCM (5 mL). The reaction mixture was stirred at 25 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (20 mL) and extracted with DCM (100 mL×2). The combined organic layer was washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 1: 0 to 0: 1). The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 1: 0 to 4: 5). Example 18 was obtained as a white solid as 1-[(1-hydroxy-5-isoquinolyl)sulfonyl]-4-methylindoline-6-carbonitrile (8 mg, 19.9 μmol, 6.30% yield). LC-MS: [M+1] 365.08.

[0225] 1 H NMR (400 MHz, MeOD): δ8.61 (d, J = 8 Hz, 1H), 8.38 (d, J = 8 Hz, 1H), 7.64 (t, J = 8 Hz, 1H), 7.56 (s, 1H), 7.37 (d, J = 7.6 Hz, 1H), 7.28 (d, J = 8 Hz, 1H), 7.21 (s, 1H), 4.07 (t, J = 8.4 Hz, 2H), 2.96 (t, J = 8.4 Hz, 2H), 2.20 (s, 3H). One proton was exchanged.

[0226] Embodiment 19

[0227]

[0228] 7-Chloro-5-((6-chloroindol-1-yl)sulfonyl)isoquinolin-1(2H)-one

[0229]

[0230] Compound 19-2: A mixture of 3,5-dichlorobenzaldehyde (25 g, 142.8 mmol), 2,2-diethoxyethylamine (83.08 mL, 571.3 mmol) in toluene (200 mL) was degassed and purged with N2 three times, and then the mixture was stirred and placed at 110 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (500 mL) and extracted with EtOAc (900 mL×2). The combined organic layer was washed with saturated brine (200 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The crude product compound 19-2 was obtained as a yellow solid, which was (E)-1-(3,5-dichlorophenyl)-N-(2,2-diethoxyethyl)methanamine (35 g, crude product). The crude product can be used in the next step without further purification.

[0231] Compound 19-3: (E)-1-(3,5-dichlorophenyl)-N-(2,2-diethoxyethyl)methamine (30 g, 103.3 mmol) was added dropwise to a solution in H2SO4 (500 mL) in DCM (100 mL) at 125 ° C. The resulting mixture was stirred at 125 ° C for 6 h under N2 atmosphere. The reaction mixture was quenched by adding ice water (200 mL) at 0 ° C, and then an aqueous NaOH solution (200 mL) was added to the mixture to adjust to pH 8. It was extracted with EtOAc (1000 mL×2). The combined organic layer was washed with saturated brine (200 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 20: 1 to 5: 1).

[0232] Compound 19-3 was obtained as a yellow solid as 5,7-dichloroisoquinoline (8 g, 40.3 mmol, 39.0% yield).

[0233] Compound 19-4: a mixture of 5,7-dichloroisoquinoline (100 mg, 504.9 μmol), diphenylmethamine (137.2 mg, 757.3 μmol), (5-diphenylphosphino-9,9-dimethyl-xanthen-4-yl)-diphenylphosphine, methanesulfonate 2-[2-(methylamino)phenyl]phenyl]palladium (485.9 mg, 504.9 μmol), tert-butylbenzene (t-BuONa) (2M, 1.26 mL) in toluene (10 mL) was degassed and purged with N2 three times, and then the mixture was stirred at 100 ° C. under N2 atmosphere for 12 h. The reaction mixture was diluted with H2O (20 mL) and extracted with DCM (80 mL×2). The combined organic layer was washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate=20:1 to 1:1).

[0234] Compound 19-4, N-(7-chloro-5-isoquinolyl)-1,1-diphenylmethanimine (40 mg, 116.6 μmol, 11.5% yield), was obtained as a yellow solid.

[0235] Compound 19-5: HCl (3M, 525.0 μL) was added to a solution of N-(7-chloro-5-isoquinolyl)-1,1-diphenylmethanamine (0.3 g, 875.0 μmol) in THF (10 mL). The mixture was stirred at 25 ° C for 2 h under N2 atmosphere. The reaction mixture was quenched by adding ice water (10 mL) at 0 ° C, after which NaOH (10 mL) was added to the mixture to adjust to pH 8. The reaction mixture was diluted with H2O (10 mL) and extracted with DCM (50 mL×2). The combined organic layer was washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 5: 1 to 1: 1). Compound 19-5 was obtained as a yellow solid, 7-chloroisoquinolin-5-amine (0.1 g, 559.8 μmol, 63.9% yield).

[0236] Compound 19-6: NaNO2 (38.6 mg, 559.8 μmol) and H2O (1 mL) were added to a solution of 7-chloroisoquinolin-5-amine (0.1 g, 559 μmol) in HCl (5 mL) at -5 °C. The mixture was stirred at -5 °C for 1 h. It was then transferred in batches to a solution of AcOH (5 mL) and CuCl (13.8 mg, 139.9 μmol) in H2O (1 mL), and the solution was bubbled with SO2 to reach saturation. The mixture was stirred at 25 °C for 2 h. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was taken up with water (20 mL) and DCM, and extracted with DCM (80 mL×2). The combined organic layers were washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The crude product compound 19-6 was obtained as 7-chloroisoquinoline-5-sulfonyl chloride (0.2 g, crude) as a yellow solid and was used in the next step without further purification.

[0237] Compound 19-7: K2CO3 (158.1 mg, 1.14 mmol) and 6-chloroindoline (128.9 mg, 839.3 μmol) were added to a solution of 7-chloroisoquinoline-5-sulfonyl chloride (0.2 g, 763.0 μmol) in acetone (10 mL). The mixture was stirred at 25 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (20 mL) and extracted with DCM (80 mL×2). The combined organic layer was washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 1: 0 to 1: 1).

[0238] Compound 19-7, 7-chloro-5-(6-chloroindolin-1-yl)sulfonyl-isoquinoline, was obtained as a yellow solid (80 mg, 210.9 μmol, 27.6% yield).

[0239] Compound 19-8: A mixture of 7-chloro-5-(6-chloroindolyl-1-yl)sulfonyl-isoquinoline (80 mg, 210.9 μmol) and m-CPBA (65.7 mg, 316.4 μmol) in DCM (5 mL) was cooled to 0 ° C, and then the mixture was stirred at 25 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (15 mL) and extracted with DCM (25 mL×2). The combined organic layer was washed with 10 mL of saturated brine, dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: EtOAc = 10: 1 to DCM: methanol = 10: 1). Compound 19-8, 7-chloro-5-(6-chloroindolin-1-yl)sulfonyl-2-oxido-isoquinolin-2-ium (50 mg, 126.5 μmol, 59.9% yield), was obtained as a yellow solid.

[0240] Compound 19-9: AC2O A mixture of 7-chloro-5-(6-chloroindol-1-yl)sulfonyl-2-oxoisoquinolin-2-ium (50 mg, 126.5 μmol) in AC2O (3 mL) was heated to 90°C. ， The mixture was then stirred at 90 ° C for 3 h under N2 atmosphere. The reaction mixture was concentrated under reduced pressure to give a residue. The crude product [7-chloro-5-(6-chloroindolyl-1-yl)sulfonyl-1-isoquinolyl]acetate (40 mg, crude) was obtained as a brown solid, which was used in the next step without further purification.

[0241] Example 19: A mixture of [7-chloro-5-(6-chloroindolyl-1-yl)sulfonyl-1-isoquinolyl]acetate (40 mg, 91.4 μmol) and KOH (7.70 mg, 137.2 μmol) in methanol (5 mL) was degassed and purged with N2 three times, and then the mixture was stirred at 25°C for 2 h under N2 atmosphere. The mixture was then stirred at 70°C for 3 h under N2 atmosphere. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (chromatographic column: Xtimate C18 150 mm x 25 mm x 5 μm; mobile phase: [A, water (10 mM NH4HCO3)-B, ACN]; B%: 50%-70%, 10 minutes). Example 19 was obtained as 7-chloro-5-(6-chloroindolin-1-yl)sulfonyl-2H-isoquinolin-1-one as a yellow solid (18 mg, 45.0 μmol, 49.2% yield). LC-MS: [M+1] 393.99.

[0242] 1H NMR (400 MHz, DMSO-d6): δ8.43 (d, J = 4.4 Hz, 1H), 8.21 (d, J = 2.8 Hz, 1H), 7.38 (d, J = 7.6 Hz, 1H), 7.33 (d, J = 1.6 Hz, 1H), 7.23 (d, J = 8 Hz, 1H), 7.08 (t, J = 7.6 Hz, 2H), 4.05 (t, J = 8.4 Hz, 2H), 2.95 (t, J = 8.4 Hz, 2H). One proton was exchanged.

[0243] Embodiment 20

[0244]

[0245] 1-((7-Chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

[0246]

[0247] Compound 20-1: K2CO3 (79.0 mg, 572.2 μmol) and indoline-6-carbonitrile (60.5 mg, 419.6 μmol) were added to a solution of 7-chloroisoquinoline-5-sulfonyl chloride (0.1 g, 381.5 μmol) in acetone (8 mL). The mixture was stirred at 25 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (20 mL) and extracted with DCM (80 mL×2). The combined organic layer was washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 0: 1). Compound 20-1 was obtained as a yellow solid, 1-[(7-chloro-5-isoquinolyl)sulfonyl]indoline-6-carbonitrile (30 mg, 81.1 μmol, 21.2% yield).

[0248] Compound 20-2: A mixture of 1-[(7-chloro-5-isoquinolyl)sulfonyl]indoline-6-carbonitrile (30 mg, 81.1 μmol), m-CPBA (26.2 mg, 121.6 μmol) in DCM (5 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 25 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (15 mL) and extracted with DCM (50 mL×2). The combined organic layer was washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 5: 1 to DCM: McOH = 10: 1). Compound 20-2, 1-(7-chloro-2-oxido-isoquinolin-2-ium-5-yl)sulfonyl indoline-6-carbonitrile (30 mg), was obtained as a yellow solid.

[0249] Compound 20-3: A mixture of 1-(7-chloro-2-oxido-isoquinolin-2-yl-5-yl)sulfonyl indoline-6-carbonitrile (30 mg, 77.7 μmol) in Ac2O (3 mL) was degassed and purged with N2 three times, and then the mixture was stirred at 100 ° C. for 12 h under N2 atmosphere. The reaction mixture was concentrated under reduced pressure to give a residue. The crude product compound 20-3, [7-chloro-5-(6-cyanoindoline-1-yl)sulfonyl-1-isoquinolinyl] acetate (20 mg, crude product) was obtained as a yellow oil, which can be used in the next step without further purification.

[0250] Example 20: A mixture of [7-chloro-5-(6-cyanoindolyl-1-yl)sulfonyl-1-isoquinolyl]acetate (20 mg, 46.7 μmol) and KOH (5.25 mg, 93.4 μmol) in methanol (5 mL) was degassed and purged with N2 three times, and then the mixture was stirred at 25 ° C. for 2 h under N2 atmosphere. The reaction mixture was diluted with H2O (10 mL) and extracted with DCM (50 mL×2). The combined organic layer was washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (chromatographic column: HUAPU C8 Extreme BDS150mmx 30mm x 5um; mobile phase: [A, watcr (10mM NH4HCO3)-B, ACN]; B%: 45%-55%, 10 minutes).

[0251] Example 20 was obtained as 1-[(7-chloro-1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrile as a white solid (2 mg, 4.7 μmol, 10.2% yield). LC-MS: [M+1] 385.03.

[0252] 1 H NMR (400 MHz, DMSO-d6): δ8.44 (d, J = 2.4 Hz, 1H), 8.32 (d, J = 2 Hz, 1H), 7.68 (s, 1H), 7.52 (d, J = 1.2 Hz, 1H), 7.45-7.36 (m, 2H), 7.04 (d, J = 8.8 Hz, 1H), 4.07 (t, J = 8.4 Hz, 2H), 3.07 (t, J = 8.4 Hz, 2H). One proton was exchanged.

[0253] Embodiment 21

[0254]

[0255] 5-((6-Chloroindolyl-1-yl)sulfonyl)-4-fluoroisoquinolin-1-ol

[0256]

[0257] Compound 21-2: At -65 ° C, to a solution of n-BuLi (2.5M, 201.8mL) in THF (100mL), a solution of 4-bromoisoquinoline (30g, 144.1mmol) in THF (100mL) was added. The mixture was stirred at -65 ° C for 30 minutes. A solution of N-(benzenesulfonyl)-N-fluoro-benzenesulfonamide (100.0g, 317.2mmol) in THF (100mL) was added at -65 ° C. The mixture was stirred at -65 ° C for 1h. The mixture was then stirred at 25 ° C for 10.5h under N2 atmosphere. The reaction mixture was diluted with H2O (500mL) and extracted with EtOAc (1500mL×2). The combined organic layer was washed with saturated brine (100mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The crude product compound 21-2 was obtained as brown oil, which was 4-fluoroisoquinoline (30 g, crude). The crude product was used in the next step without further purification.

[0258] Compound 21-3: KNO3 (6.05 g, 59.8 mmol) was added to a solution of 4-fluoroisoquinoline (8 g, 54.3 mmol) in H2SO4 (50 mL). The mixture was stirred at 25 ° C for 12 h under N2 atmosphere. The reaction mixture was quenched by adding ice water (100 mL) at -15 ° C, and then an aqueous NaOH solution (50 mL) was added to the mixture to adjust to pH 8. It was filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 1: 1). Compound 21-3 was obtained as a yellow solid, which was 4-fluoro-5-nitro-isoquinoline (5 g, 26.0 mmol, 47.8% yield).

[0259] Compound 21-4: A mixture of 4-fluoro-5-nitro-isoquinoline (5 g, 26.0 mmol), NH4Cl (11.1 g, 208.1 mmol), Zn (13.6 g, 208.1 mmol) in THF (20 mL) and H2O (10 mL) was degassed and purged with N2 3 times, and then the mixture was stirred at 25 ° C. for 5 h under N2 atmosphere. The reaction mixture was filtered and the filtrate was concentrated. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 1: 1). Compound 21-4 was obtained as a yellow solid, which was 4-fluoroisoquinoline-5-amine (3 g, 18.5 mmol, 71.1% yield).

[0260] Compound 21-5: To a solution of 4-fluoroisoquinoline-5-amine (3 g, 18.5 mmol) in HCl (10 mL) was added NaNO3 (1.57 g, 18.5 mmol) and H2O (2 mL) at -15 ° C. The mixture was stirred at -15 ° C for 1 h under N2 atmosphere. It was then transferred to a solution of AcOH (10 mL) consisting of CuCl (457.8 mg, 4.62 mmol) and H2O (2 mL), which was saturated with bubbling SO2. The mixture was stirred at -5 ° C for 2 h. The reaction mixture was concentrated under reduced pressure to obtain a residue. The crude product compound 21-5 was obtained as a yellow solid, which was 4-fluoroisoquinoline-5-sulfonyl chloride (1.1 g, crude product), which was used in the next step without further purification.

[0261] Compound 21-6: A mixture of 4-fluoroisoquinoline-5-sulfonyl chloride (0.5 g, 2.04 mmol), 6-chloroindoline (250.1 mg, 1.63 mmol), TEA (424.9 μL, 3.05 mmol) in DCM (10 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 25 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (30 mL) and extracted with DCM (100 mL×2). The combined organic layer was washed with saturated brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether ethyl acetate = 10: 1 to 1: 1). Compound 21-6, 5-(6-chloroindolin-1-yl)sulfonyl-4-fluoro-isoquinoline (0.2 g, 551.2 μmol), was obtained as a yellow solid.

[0262] Compound 21-7: A mixture of 5-(6-chloroindolyl-1-yl)sulfonyl-4-fluoro-isoquinoline (0.2 g, 551 μmol) and m-CPBA (142.6 mg, 826.9 μmol) in DCM (10 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 25 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (20 mL) and extracted with DCM (100 mL×2). The combined organic layer was washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 5: 1 to DCM: methanol = 10: 1). Compound 21-7, 5-(6-chloroindolin-1-yl)sulfonyl-4-fluoro-2-oxido-isoquinolin-2-ium (70 mg, 184 μmol, 33.5% yield), was obtained as a white solid.

[0263] Example 21: A mixture of 5-(6-chloroindoline-1-yl)sulfonyl-4-fluoro-2-oxido-isoquinolin-2-ium (70 mg, 184.7 μmol) in Ac2O (10 mL) was degassed and purged with N2 three times, and then the mixture was stirred at 110 ° C for 8 h under N2 atmosphere. The reaction mixture was diluted with H2O (20 mL) and extracted with DCM (80 mL×2). The combined organic layer was washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 0: 1). The residue was purified by preparative HPLC (column: HUAPUC8Extreme BDS 150 mm x 30 mm x 5 μm; mobile phase: [A, water (10 mM NH4HCO3)-B, ACN]; B: 35%-55%, 10 minutes). Example 21 was obtained as a white solid, 5-(6-chloroindolyl-1-yl)sulfonyl-4-fluoro-isoquinolin-1-ol (5 mg, 12.2 μmol, 6.61% yield). LC-MS: [M+1] 378.81.

[0264] 1 H NMR (400 MHz, MeOD): δ9.10 (s, HI), 8.74 (d, J = 7.6 Hz, 1H), 8.23 ​​(d, J = 7.6 Hz, 1H), 7.97 (d, J = 6.8 Hz, 1H), 7.81 (t, J = 7.6 Hz, 1H), 7.35 (d, J = 8 Hz, 1H), 7.12-7.15 (m, 1H), 4.16 (t, J = 8.8 Hz, 2H), 3.23 (t, J = 8.4 Hz, 2H). One proton was exchanged.

[0265] Embodiment 22

[0266]

[0267] 1-((1-Hydroxy-4-methylisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

[0268]

[0269] Compound 22-1: To a mixture of KNO3 (21.3 g, 211.4 mmol) in H2SO4 (600 mL) was added 4-bromoisoquinoline (40 g, 192.2 mmol) at 0°C, and the mixture was stirred at 25°C for 12 h under N2 atmosphere. The reaction mixture was quenched by adding ice water (3000 mL), and then NH3.H2O was added to the mixture to adjust to pH 8. The mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was washed with methanol (500 mL) and then filtered. The solid was dissolved with EtOAc (500 mL) and H2O (1000 mL). The organic layer was concentrated under reduced pressure to obtain a residue.

[0270] Compound 22-1 was obtained as a yellow solid as 4-bromo-5-nitro-isoquinoline (75 g, 296.3 mmol, 77.0% yield).

[0271] Compound 22-2: To a mixture of 4-bromo-5-nitro-isoquinoline (55 g, 217.3 mmol) in dioxane (500 mL) and H2O (50 mL) were added methylboric acid (26.02 g, 434.6 mmol), Cs2CO3 (141.6 g, 434.6 mmol) and Pd(dppf)Cl2 (15.90 g, 21.73 mmol) in batches. The mixture was heated to 110°C and stirred for 12 h. The reaction mixture was filtered. The filtrate was diluted with H2O (1.2 L) and extracted with EtOAc (800 mL×2). The combined organic layers were washed with brine (1.2 L), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 1:0 to 3:1). Compound 22-2 was obtained as a yellow solid as 4-methyl-5-nitro-isoquinoline (36 g, 191.3 mmol, 88.0% yield).

[0272] Compound 22-3: To a solution of 4-methyl-5-nitro-isoquinoline (33 g, 175.3 mmol) in THF (500 mL) and H2O (250 mL) was added Zn (91.7 g, 1.40 mol) and NH4Cl (75.0 g, 1.40 mol). The mixture was stirred at 25 ° C for 12 h. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 1: 0 to 0: 1). Compound 22-3 was obtained as a yellow solid, which was 4-methylisoquinoline-5-amine (20 g, 113.8 mmol, 64.8% yield).

[0273] Compound 22-4: Solution 1: To a mixture of 4-methylisoquinolin-5-amine (3 g, 18.9 mmol) in HCl (30 mL) was added NaNO2 (1.31 g, 18.9 mmol) and H2O (6 mL) in batches at -5°C under N2. The mixture was stirred at -5°C for 1 h. Solution 2: SO2 was bubbled into a solution of CuCl (469.3 mg, 4.74 mmol) in HOAc (30 mL) and H2O (6 mL) at 0°C for 30 minutes. The two solutions were mixed and stirred at 25°C for 1.5 h. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with H2O (300 mL) and extracted with DCM (200 mL×3). The combined organic layers were washed with brine (300 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. Compound 22-4 was obtained as a yellow solid, 4-methylisoquinoline-5-sulfonyl chloride (2 g, crude), which was used directly without purification.

[0274] Compound 22-5: K2CO3 (3.43 g, 24.8 mmol) and indoline-6-carbonitrile (1.19 g, 8.27 mmol) were added to a solution of 4-methylisoquinoline-5-sulfonyl chloride (2 g, 8.27 mmol) in acetone (30 mL). The mixture was stirred at 25 ° C for 12 h. The mixture was heated to 50 ° C and stirred for 2 h. The reaction mixture was diluted with H2O (200 mL) and extracted with DCM (200 mL×2). The combined organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 1: 0 to 0: 1). Compound 22-5 was obtained as a yellow solid, which was 1-[(4-methyl-5-isoquinolyl)sulfonyl]indoline-6-carbonitrile (1.1 g, 2.52 mmol, 30.4% yield).

[0275] Compound 22-6: At 0°C, m-CPBA (1.44 g, 6.93 mmol) was added to a solution of 1-[(4-methyl-5-isoquinolinyl)sulfonyl]indoline-6-carbonitrile (1.1 g, 3.15 mmol) in DCM (30 mL). The mixture was warmed to 25°C and stirred for 12 h. The reaction mixture was directly purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 1: 0, ethyl acetate: EtOH = 1: 1). Compound 22-6 was obtained as a yellow solid, which was 1-(4-methyl-2-oxidized-isoquinolin-2-ium-5-yl)sulfonylindoline-6-carbonitrile (1 g, 2.54 mmol, 80.7% yield).

[0276] Example 22: A mixture of 1-(4-methyl-2-oxido-isoquinolin-2-ium-5-yl)sulfonyl indoline-6-carbonitrile (1 g, 2.74 mmol) in Ac2O (20 mL) was heated to 120°C and stirred for 12 h. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was dissolved in DMF (20 mL) and filtered. The filtrate was purified by preparative HPLC (chromatographic column: WatersXbridge Prep OBD C18 150 mm x 40 mm x 10 μm; mobile phase: [A, water (10 mM NH4HCO3)-B, ACN]; B%: 20%-50%, 11 minutes) and concentrated. The product was added to EtOH (2 mL) and filtered to give 1-[(1-hydroxy-4-methyl-5-isoquinolinyl)sulfonyl]indoline-6-carbonitrile (19 mg) as a light yellow solid. LC-MS: [M+1] 365.08.

[0277] 1 H NMR (400MHz, DMSO-d6): δ11.71(br,1H),8.59(d,J=6.4Hz,1H),7.89(d,J=6.4Hz,1H),7 .59~7.51(m,3H),7.25(s,2H),4.22(t,J=8.8Hz,2H),3.4(t,J=8.4Hz,2H),2.61(s,3H).

[0278] Embodiment 23

[0279]

[0280] 1-((1-Hydroxy-4-methylisoquinolin-5-yl)sulfonyl)-4-methylindoline-6-carbonitrile

[0281]

[0282] Compound 23-1: To a solution of 4-methylisoquinoline-5-sulfonyl chloride (100 mg, 413.7 μmol) in acetone (5 mL) was added K2CO3 (171.5 mg, 1.24 mmol) and 4-methylindoline-6-carbonitrile (52.36 mg, 331.0 μmol). The mixture was stirred at 25 ° C for 12 h. The mixture was heated to 50 ° C and stirred for 6 h. The reaction mixture was diluted with H2O (100 mL) and extracted with DCM (100 mL×2). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 1: 0 to 0: 1).

[0283] Compound 23-1, 4-methyl-1-[(4-methyl-5-isoquinolyl)sulfonyl]indoline-6-carbonitrile (50 mg, crude), was obtained as a yellow solid.

[0284] Compound 23-2: m-CPBA (62.9 mg, 302.7 μmol) was added to a solution of 4-methyl-1-[(4-methyl-5-isoquinolinyl)sulfonyl]indoline-6-carbonitrile (50 mg, 137 μmol) in DCM (10 mL) at 0°C. The mixture was heated to 25°C and stirred for 2 h. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 1:0, ethyl acetate: methanol = 5:1). Compound 23-2 was obtained as a yellow solid, which was 4-methyl-1-(4-methyl-2-oxidized-isoquinolin-2-ium-5-yl)sulfonyl-indoline-6-carbonitrile (40 mg, crude product).

[0285] Example 23: A mixture of 4-methyl-1-(4-methyl-2-oxido-isoquinolin-2-ium-5-yl)sulfonyl-indoline-6-carbonitrile (40 mg, 105.4 μmol) in Ac2O (5 mL) was heated to 100°C and stirred for 12 h. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was purified by preparative HPLC (column: Luna C18 100 mm x 30 mm x 5 μm; mobile phase: [A, water (0.1% TFA)-B, ACN]; B%: 30%-55%, 12 minutes). Example 23 was obtained as a brown solid as 1-[(1-hydroxy-4-methyl-5-isoquinolinyl)sulfonyl]-4-methyl-indoline-6-carbonitrile (1.6 mg, 3.97 μmol, 3.76% yield). LC-MS: [M+1] 379.10.

[0286] 1 H NMR (400 MHz, DMSO-d6): δ11.71 (br, 1H), 8.59 (d, J=8 Hz, 1H), 7.88 (d, J=7.6 Hz, 1H), 7.53 (t, J=8 Hz, 1H), 7.39 (s, 1H), 7.26-7.25 (m, 1H), 7.09 (s, 1H), 4.22 (t, J=8 Hz, 2H) 2.61 (s, 6H) (2H may overlap with the water peak).

[0287] Embodiment 24

[0288]

[0289] 5-((6-Chloroindolyl-1-yl)sulfonyl)-4-methylisoquinolin-1-ol

[0290]

[0291] Compound 24-1: K2CO3 (857.7 mg, 6.21 mmol) and 6-chloroindoline (317.8 mg, 2.07 mmol) were added to a solution of 4-methylisoquinoline-5-sulfonyl chloride (500 mg, 2.07 mmol) in acetone (10 mL). The mixture was stirred at 25 ° C for 12 h. The reaction mixture was filtered and the filtrate was diluted with H2O (200 mL) and extracted with EtOAc (150 mL×2). The combined organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 1: 0 to 0: 1). Compound 24-1 was obtained as a yellow solid, which was 5-(6-chloroindoline-1-yl)sulfonyl-4-methyl-isoquinoline (400 mg, 947.4 μmol, 45.80% yield).

[0292] Compound 24-2: At 0°C, m-CPBA (579.3 mg, 2.79 mmol) was added to a solution of 5-(6-chloroindolyl-1-yl)sulfonyl-4-methyl-isoquinoline (400 mg, 1.11 mmol) in DCM (10 mL). The mixture was heated to 25°C and stirred for 2.5 h. The reaction mixture was directly purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 1:0, ethyl acetate: methanol = 5:1). Compound 24-2 was obtained as a yellow solid, which was 5-(6-chloroindolyl-1-yl)sulfonyl-4-methyl-2-oxidized-isoquinolin-2-ium (300 mg, crude product).

[0293] Example 24: A mixture of 5-(6-chloroindol-1-yl)sulfonyl-4-methyl-2-oxido-isoquinolin-2-ium (300 mg, 800.3 μmol) in Ac2O (10 mL) was heated to 100°C and stirred for 12 h. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was purified by preparative HPLC (chromatographic column: Waters Xbridge Prep OBD C18 150 mm x40 mm x 10 μm; mobile phase: [A, water (10 mM NH4HCO3)-B, ACN]; B%: 30%-55%, 11 minutes). Methanol (2 mL) was added, the mixture was filtered and concentrated under reduced pressure to give compound 24, 5-(6-chloroindol-1-yl)sulfonyl-4-methyl-isoquinolin-1-ol (6 mg, 15.2) μmol, 40.7% yield) as a yellow solid. LC-MS: [M+1] 374.05.

[0294] 1H NMR (400 MHz, DMSO-d6): δ 11.70 (br, 1H), 8.59 (d, J = 4.4 Hz, 1H), 7.86 (d, J = 7.2 Hz, 1H), 7.54 (t, J = 7.6 Hz, 1H), 7.37 (d, J = 8 Hz, 1H), 7.25 (d, J = 4.8 Hz 1H), 7.09 (d, J = 8.8 Hz, 1H), 6.98 (s, 1H), 4.2 (t, J = 8 Hz, 1H), 3.26 (m, 2H, overlapped with solvent peak), 2.61 (s, 3H). One proton was exchanged.

[0295] Embodiment 25

[0296]

[0297] 5-((6-Chloroindol-1-yl)sulfonyl)phthalazin-1(2H)-one

[0298]

[0299] Compound 25-2: A mixture of 4-nitroisoindoline-1,3-dione (25 g, 130.1 mmol), NaBH4 (11.13 g, 294.0 mmol) in methanol (250 mL) was degassed and purged with N2 three times. The mixture was stirred at 25 ° C for 2 h under N2 atmosphere. The solution was acidified with 20% HCl (100 mL), and the reaction mixture was concentrated under reduced pressure to obtain a residue. The dried residue was treated with acetone. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 0: 1). Compound 25-2 was obtained as a yellow solid, which was 3-hydroxy-4-nitro-isoindoline-1-one (15 g, 59% yield).

[0300] Compound 25-3: A mixture of 3-hydroxy-4-nitro-isoindolin-1-one (10 g, 51.5 mmol) in HCl (70 mL, 20% aqueous solution) was degassed and purged with N2 3 times. The mixture was stirred at 90 ° C for 12 h under N2 atmosphere. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 1: 1). Compound 25-3 was obtained as a yellow solid, which was 3-hydroxy-4-nitro-3H-isobenzofuran-1-one (10 g, crude product), which was used without further purification.

[0301] Compound 25-4: A mixture of 3-hydroxy-4-nitro-3H-isobenzofuran-1-one (10 g, 51.2 mmol), N2H4.H2O (9.05 g, 153.7 mmol) in EtOH (200 mL) was degassed and purged with N2 three times. The mixture was stirred at 80 ° C for 5 h under N2 atmosphere. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate to DCM: methanol = 10: 1 to 10: 1).

[0302] Compound 25-4, 5-nitro-2H-phthalazin-1-one, was obtained as a yellow solid (6 g, 31.3 mmol, 61.2% yield).

[0303] Compound 25-5: A mixture of 5-nitro-2H-phthalazin-1-one (6 g, 31.3 mmol), Pd / C (3 g, 10 wt%) in methanol (100 mL) was degassed and purged with H2 three times, and then the mixture was stirred under H2 atmosphere at 25°C and a pressure of 15 psi for 12 h. The reaction mixture was filtered and the filtrate was concentrated. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate to DCM: methanol = 10: 1 to 10: 1).

[0304] Compound 25-5, 5-amino-2H-phthalazin-1-one, was obtained as a yellow solid (3 g, 18.6 mmol, 59.3% yield).

[0305] Compound 25-6: To a solution of 5-amino-2H-phthalazin-1-one (3 g, 18.61 mmol) in HCl (20 mL) was added NaNO2 (1.28 g, 18.61 mmol) and H2O (5 mL) at -5°C. The mixture was stirred at -5°C for 1 h. It was then transferred in batches to a solution of AcOH (15 mL) with CuCl (460.7 mg, 4.65 mmol) and H2O (5 mL), which was saturated by bubbling SO2. The mixture was stirred at 25°C for 2 h under N2 atmosphere. The reaction mixture was concentrated under reduced pressure to give a residue. The crude product of compound 25-6 was obtained as a yellow solid, 1-oxo-2H-phthalazine-5-sulfonyl chloride (1.5 g, 6.13 mmol, 32.9% yield), which was used in the next step without further purification.

[0306] Example 25: TEA (853.3 uL, 6.13 mmol) and 6-chloroindoline (502.2 mg, 3.27 mmol) were added to a solution of 1-oxo-2H-phthalazine-5-sulfonyl chloride (1 g, 4.09 mmol) in DCM (15 mL). The mixture was stirred at 25 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (50 mL) and extracted with DCM (150 mL×2). The combined organic layer was washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 0: 1). Compound 25 was obtained as a yellow solid, which was 5-(6-chloroindoline-1-yl)sulfonyl-2H-phthalazine-1-one (0.15 g, 373.3 μmol, 9.13% yield). LC-MS: [M+1] 361.03.

[0307] 1 H NMR (400MHz, DMSO-d6): δ13.08(s,1H),8.78(s,1H),8.56(d,J=8Hz,1H),8.46(d,J=8Hz,1H),8.02(t,J=8Hz ,1H),7.39(s,1H),7.22(d,J=8.4Hz,1H),7.09(d,J=8.4Hz,1H),4.06(t,J=8Hz,2H),2.90(t,J=8.4Hz,2H).

[0308] Embodiment 26

[0309]

[0310] 1-((1-Oxo-1,2-dihydrophthalazin-5-yl)sulfonyl)indoline-6-carbonitrile

[0311]

[0312] Example 26: TEA (426uL, 3.07mmol) and indoline-6-carbonitrile (235mg, 1.63mmol) were added to a solution of 1-oxo-2H-phthalazine-5-sulfonyl chloride (0.5g, 2.04mmol) in DCM (15mL). The mixture was stirred at 25°C for 12h under N2 atmosphere. The reaction mixture was diluted with H2O (20mL) and extracted with EtOAc (80mL×2). The combined organic layers were washed with saturated brine (20mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 10: 1 to 0: 1). Compound 26 was obtained as a yellow solid, which was 1-[(1-oxo-2H-phthalazine-5-yl)sulfonyl]indoline-6-carbonitrile (30mg, 81.6μmol, 3.99% yield). LC-MS: [M+1] 352.06.

[0313] 1 H NMR (400MHz, DMSO-d6): δ13.09(s,1H),8.81(s,1H),8.56(d,J=8Hz,2H),8.01(t,J=8Hz,1H),7.72 (s, 1H), 7.52 (d, J = 9.2Hz, 1H), 7.41 (d, J = 7.6Hz, 1H), 4.08 (t, J = 8.4Hz, 2H), 3.03 (t, J = 8.4Hz, 2H).

[0314] Embodiment 27

[0315]

[0316] 1-((4-Chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

[0317]

[0318] Compound 27-2. To a mixture of 4-chloroisoquinoline (5g, 30.56mmol, 1eq) in H2SO4 (20mL) was added KNO3 (3.40g, 33.62mmol, 1.1eq) at 0°C and N2. The mixture was stirred at 15°C for 12h. The reaction mixture was added to ice-cold water (100ml), and then a saturated NaOH solution was added until the pH was adjusted to pH = 10. The mixture was extracted with EtOAc (200mL*2). The combined organic layer was washed with brine (200mL), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was washed with a mixed solvent (50mL) of petroleum ether / ethyl acetate = 1 / 1. The mixture was filtered and concentrated under reduced pressure to obtain a residue. Compound 27-2 was obtained as a yellow solid, which was 4-chloro-5-nitro-isoquinoline (4g, 19.18mmol, 62.74% yield).

[0319] Compound 27-3: At 0°C, to a mixture of 4-chloro-5-nitro-isoquinoline (4 g, 19.18 mmol, 1 eq) and NH4Cl (8.21 g, 153.40 mmol, 8 eq) in THF (50 mL) and H2O (25 mL) solution) in Zn (10.03 g, 153.40 mmol, 8 eq) was added, and the resulting mixture was stirred at 15°C for 3 h. The reaction mixture was filtered, and the filtrate was added to H2O (200 mL) and extracted with EtOAc (150 mL*2). The combined organic layer was washed with brine (200 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). Compound 27-3, 4-chloroisoquinolin-5-amine, was obtained as a light yellow solid (1.5 g, 8.40 mmol, 43.79% yield).

[0320] Preparation of solution 1: To a solution of 4-chloroisoquinolin-5-amine (1.5 g, 8.40 mmol, 1 eq) in concentrated hydrochloric acid (15 mL) was added NaNO2 (579.45 mg, 8.40 mmol, 1 eq) portionwise at -5°C under N2. The mixture was stirred at -5°C for 1 h.

[0321] Preparation of solution 2: SO2 was bubbled into a solution of CuCl (207.85 mg, 2.10 mmol, 0.25 eq) in HOAc (15 mL) and H2O (3 mL) at 0°C for 30 min.

[0322] Compound 27-4: Solution 2 was slowly added to solution 1. The mixture was stirred at 15 ° C for 2 h. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with H2O (500 mL) and extracted with DCM (300 mL*3). The combined organic layer was washed with brine (500 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue, which was used without further purification. Compound 27-4 was obtained as a yellow solid, which was 4-chloroisoquinoline-5-sulfonyl chloride (1.5 g, crude product).

[0323] Compound 27-5: K2CO3 (1.58 g, 11.45 mmol, 2 eq) was added in batches to a mixture of 4-chloroisoquinoline 5-sulfonyl chloride (1.5 g, 5.72 mmol, 1 eq) and indoline-6-carbonitrile (825.06 mg, 5.72 mmol, 1 eq) in DMF (20 mL) at 15 ° C and N2. The mixture was stirred at 15 ° C for 12 h. The reaction mixture was diluted with H2O (200 mL) and extracted with EtOAc (150 ml * 3). The combined organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). Compound 27-5 was obtained as a yellow solid, which was 1-[((4-chloro-5-isoquinolyl)sulfonyl]indoline-6-carbonitrile (600 mg, crude product).

[0324] Compound 27-6: To a solution of 1-[(4-chloro-5-isoquinolyl)sulfonyl]indoline-6-carbonitrile (600 mg, 1.62 mmol, 1 eq) in DCM (20 mL) was added m-CPBA (769.93 mg, 3.57 mmol), purity 80%, 2.2 eq). The mixture was stirred at 15 ° C for 12 h. The reaction mixture was directly purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to ethyl acetate: EtOH = 3: 1). The crude product of compound 27-6 obtained was triturated with methanol (50 mL), filtered, and used in the next step without further purification.

[0325] Example 27: A mixture of 1-(4-chloro-2-oxido-isoquinolin-2-ium-5-yl)sulfonyl indoleline-6-carbonitrile (150 mg, 388.78 umol, 1 eq) in Ac2O (15 mL) was heated to 100°C and stirred for 12 h. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was purified by preparative HPLC (chromatographic column: Xtimate C18 150*25mm*5μm; mobile phase: [water (10mM NH4HCO3)-ACN]; B%: 40%-70%, 10 minutes). The residue was then re-purified by preparative HPLC (column: Xtimate C18 150*25mm*5μm; mobile phase: [water (10mM NH4HCO3)-ACN]; B%: 30%-55%, 8 minutes). Example 27 was obtained as a yellow solid as 1-[(4-chloro-1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrile (4 mg, 9.63 umol, 2.48% yield, 92.93% purity). LCMS (ESI+) m / z: 383.0.

[0326] 1 H NMR (400MHz, DMSO-d6): 8.55 (d, J = 8Hz, 1H), 7.98 (d, J = 7.6Hz, 1H), 7.63-7.46 (m, 4H), 7.21 (s, 1H), 4.15 (t, J = 8.6Hz, 2H), 3.34 (t, J = 8.8Hz, 2H).

[0327] Embodiment 28

[0328]

[0329] 1-((1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-2,3-dihydro-1H-pyrrolo[3,2-c]pyridine-6-carbonitrile

[0330]

[0331] Compound 28-2: At 20 ° C, to a solution of 6-chloro-2,3-dihydro-1H-pyrrolo[3,2-c]pyridine (1.6 g, 10.35 mmol, 1 eq) in DCM (20 mL), Boc2O (6.78 g, 31.05 mmol, 7.13 mL, 3 eq), TEA (2.09 g, 20.70 mmol, 2.88 mL, 2 eq) and DMAP (1.26 g, 10.35 mmol, 1 eq) were added. The mixture was heated to 50 ° C and stirred for 12 h. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). Compound 28-2 was obtained as tert-butyl-6-chloro-2,3-dihydropyrrolo[3,2-c]pyridine-1-carboxylate (1.5 g, 5.89 mmol, 56.90% yield) as a colorless oil.

[0332] Compound 28-3: Pd(dppf)Cl2.CH2Cl2 (2.40 g, 2.94 mmol, 0.5 eq) was added to a solution of tert-butyl-6-chloro-2,3-dihydropyrrolo[3,2-c]pyridine-1-carboxylate (1.5 g, 5.89 mmol, 1 eq) in methanol (50 mL) and DIPEA (5 mL) at 15 °C. The suspension was degassed under vacuum and purged with CO several times. The mixture was stirred at 80 °C under CO (50 psi) for 12 h. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). Compound 28-3, O-1-tert-butyl-O-6-methyl-2,3-dihydropyrrolo[3,2-c]pyridine-1,6-dicarboxylate, was obtained as a red solid (1.5 g, 4.31 mmol, 73.22% yield, 80% purity).

[0333] Compound 28-4: NaOH (676.96 mg, 16.92 mmol, 3 eq) was added to a solution of O1-tert-butyl O6-methyl 2,3-dihydropyrrolo[3,2-c]pyridine-1,6-dicarboxylate (1.57 g, 5.64 mmol, 1 eq) in methanol (20 mL) and H2O (4 mL). The mixture was stirred at 15°C for 30 minutes. The reaction mixture was concentrated under reduced pressure to remove methanol. A 12M HCl solution was then added to the remaining solution until pH = 3-4, and the mixture was extracted with EtOAc (100 mL×2). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue that was used without further purification. Compound 28-4 was obtained as a yellow solid, which was 1-tert-butoxycarbonyl-2,3-dihydropyrrolo[3,2-c]pyridine-6-carboxylic acid (700 mg, crude).

[0334] Compound 28-5: HATU (1.40 g, 3.69 mmol, 1.5 eq), NH4Cl (526.24 mg, 9.84 mmol, 4 eq) and DIPEA (1.59 g, 12.30 mmol, 2.14 mL, 5 eq) were added to a solution of 1-tert-butoxycarbonyl-2,3-dihydropyrrolo[3,2-c]pyridine-6-carboxylic acid (650 mg, 2.46 mmol, 1 eq) in DMF (10 mL). The mixture was stirred at 15 ° C for 12 h. The reaction mixture was added to H2O (150 mL) and extracted with EtOAc (80 mL*2). The combined organic layer was washed with brine (150 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). Compound 28-5 was obtained as tert-butyl-6-carbamoyl-2,3-dihydropyrrolo[3,2-c]pyridine-1-carboxylate as a white solid (300 mg, 46% yield).

[0335] Compound 28-6: To a solution of tert-butyl-6-carbamoyl-2,3-dihydropyrrolo[3,2-c]pyridine-1-carboxylate (250 mg, 949.52 μmol, 1 eq) in THF (10 mL), pyridine (195.28 mg, 2.47 mmol, 199.26 uL, 2.6 eq) and TFAA (518.51 mg, 2.47 mmol, 343.38 mL, 2.6 eq) were added. The mixture was stirred at 15 ° C for 2 h. The reaction mixture was added to H2O (100 mL) and extracted with EtOAc (50 mL*2). The combined organic layer was washed with brine (100 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 1:1). Compound 28-6, tert-butyl-6-cyano-2,3-dihydropyrrolo[3,2-c]pyridine-1-carboxylate (250 mg, crude), was obtained as a light yellow solid.

[0336] Compound 28-7: A solution of tert-butyl-6-cyano-2,3-dihydropyrrolo[3,2-c]pyridine-1-carboxylate (250 mg, 1.02 mmol, 1 eq) in HCl / EtOAc (10 mL) was stirred at 15 ° C for 12 h. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was used without further purification. Compound 28-7 was obtained as an off-white solid, which was 2,3-dihydro-1H-pyrrolo[3,2-c]pyridine-6-carbonitrile (160 mg, crude, HCl).

[0337] Compound 28-8: A mixture of 1-hydroxyisoquinoline-5-sulfonic acid (1.00 g, 4.44 mmol, 1 eq) in SOCl2 (10 mL) and DMF (0.1 mL) was heated to 40 ° C and stirred for 1 h. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with H2O (100 mL) and extracted with DCM (70 mL*2). The combined organic layer was washed with brine (100 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). Compound 28-8 was obtained as a yellow solid, which was 1-oxo-2H-isoquinoline-5-sulfonyl chloride (300 mg, crude product), which was used directly in the next step.

[0338] Example 28: To a solution of 2,3-dihydro-1H-pyrrolo[3,2-c]pyridine-6-carbonitrile (37.53 mg, 258.55 μmol, 0.7 eq) in DMF (7 mL) was added NaH (59.10 mg, 1.48 mmol, 78.02 μL, 60% purity, 4 eq) at 15°C. The mixture was stirred at this temperature for 30 minutes, and then 1-oxo-2H-isoquinoline-5-sulfonyl chloride (90 mg, 369.36 μmol, 1 eq) was added at 15°C. The resulting mixture was stirred at 15°C for 1.5 h. The reaction mixture was quenched by adding H2O (20 mL), then diluted with H2O (40 mL) and extracted with EtOAc (20 mL*2). The combined organic layers were washed with brine (60 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (chromatographic column: XtimateC18 150*25mm*5μm; mobile phase: [water (10mM NH4HCO3)-ACN]; B%: 20%-50%, 8 minutes). Example 28 was obtained as a white solid, 1-[(1-oxo-2H-isoquinolin-5-yl)sulfonyl]-2,3-dihydropyrrolo[3,2-c]pyridine-6-carbonitrile (3 mg, 98.76% purity). LCMS (ESI+) / mz: 353.0.

[0339] 1 H NMR (400MHz, DMSO-d6): δ11.74(br,1H),8.56(d,J=8Hz,1H),8.51~8.46(m,2H),7.75(s,1H),7.67(t ,J=8Hz,1H),7.45~7.37(m,1H),7.11(d,J=7.6Hz,1H),4.10(t,J=8.8Hz,2H),3.22(t,J=8.6Hz,2H).

[0340] Embodiment 29

[0341]

[0342] 4-Methoxy-1-((1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

[0343]

[0344] Compound 29-2: To a solution of 4-methoxy-1H-indole-6-carbonitrile (230 mg, 1.34 mmol, 1 eq) in DCM (8 mL), triethylsilane (776.63 mg, 6.68 mmol, 1.07 mL, 5 eq) was added. The reaction was then cooled to 0 ° C, and TFA (4 mL) was added dropwise at 0 ° C. The mixture was heated to 50 ° C and stirred for 12 h. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate=1 / 0 to 0 / 1) to obtain compound 29-2 as a light yellow solid, which was 4-methoxyindoleline-6-carbonitrile (100 mg, 40% yield).

[0345] Example 29: To a solution (10 mL) of 1-oxo-2H-isoquinoline-5-sulfonyl chloride (150 mg, 615.60 μmol, 1 eq) and 4-methoxyindoline-6-carbonitrile (32.17 mg, 184.68 μmol, 0.3 eq) in DCM was added TEA (207.64 mg, 1.23 mmol, 285.61 μL, 60% purity, 2 eq). The mixture was stirred at 15°C for 12 h. The reaction mixture was added to H2O (50 mL) and extracted with EtOAc (30 mL*2). The combined organic layers were concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (chromatographic column: Nano-micro Kromasil C18 100*40 mm*10 μm; mobile phase: [water (0.1% TFA)-ACN]; B%: 30%-56%, 10 minutes). The residue was purified by preparative HPLC (column: Xtimate C18 150*25mm*5μm; mobile phase: [water (10mM NH4HCO3)-ACN]; B%: 25%-55%, 8 minutes). Example 29 was obtained as a white solid, 4-methoxy-1-[(1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrile (2 mg, 96.64% purity). LCMS (ESI+) m / z: 382.0.

[0346] 1H NMR (400MHz, DMSO-d6): δ11.68(br,1H),8.52(d,J=8Hz,1H),8.34(d,J=7.6Hz,1H),7.64(t,J=8Hz,1H),7.38(d,J=7. 6Hz, 1H), 7.26 (s, 1H), 7.21 (s, 1H), 7.13 (d, J = 7.6Hz, 1H), 4.07 (t, J = 8.6Hz, 2H), 3.82 (s, 3H), 2.99 (t, J = 8.6Hz, 2H).

[0347] Examples 30 and 31

[0348]

[0349] N-(1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-6-cyanoindolin-4-yl)acetamide

[0350]

[0351] 4-Amino-1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoleline-6-carbonitrile

[0352]

[0353] Compound 30-2: Under N2, a solution of 4-nitro-1H-indole-6-carbonitrile (1.00 g, 5.34 mmol, 1 eq) in EtOH (20 mL) was added with a solution of Pd / C (1.00 g, 106.86 μmol, 5% wt%). The suspension was degassed under vacuum and purged with H2 several times. At 15 ° C, the mixture was stirred under H2 (15 psi) for 1.5 h. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give a residue, which was used without further purification. Compound 30-2 was obtained as a yellow solid, 4-amino-1H-indole-6-carbonitrile (700 mg, 4.45 mmol, 83.35% yield).

[0354] Compound 30-3: TEA (1.35 g, 13.36 mmol, 1.86 mL, 3 eq) and acetyl chloride (419.54 mg, 5.34 mmol, 381.40 μL, 1.2 eq) were added to a solution of 4-amino-1H-indole-6-carbonitrile (700 mg, 4.45 mmol, 1 eq) in DCM (15 mL). The mixture was stirred at 15 ° C for 2 h. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was purified twice by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). Compound 30-3 was obtained as a light yellow solid, N-(6-cyano-1H-indole-4-yl)acetamide (600 mg, 3.01 mmol, 67.63% yield).

[0355] Compound 30-4: To a solution of N-(6-cyano-1H-indol-4-yl)acetamide (600 mg, 3.01 mmol, 1 eq) in DCM (20 mL) was added TESH (3.50 g, 30.12 mmol, 4.81 mL, 10 eq). The reaction was then cooled to 0 ° C, and TFA (10 mL) was added dropwise at 0 ° C. The resulting mixture was stirred at 15 ° C for 12 h. The mixture was heated to 50 ° C and stirred for 5 h. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate=1 / 0 to 0 / 1).

[0356] Compound 30-4 was obtained as a light yellow solid as N-(6-cyanoindolin-4-yl)acetamide (400 mg, 1.99 mmol, 66.00% yield).

[0357] Example 30: TEA (332.22 mg, 3.28 mmol, 456.98 uL, 4 eq) and N-(6-cyanoindolin-4-yl)acetamide (82.58 mg, 410.40 umol, 0.5 eq) were added to a solution of 1-oxo-2H-isoquinoline-5-sulfonyl chloride (200 mg, 820.79 umol, 1 eq) in DCM (10 mL). The mixture was stirred at 15 ° C for 12 h. The reaction mixture was added to H2O (150 mL) and extracted with EtOAc (80 mL*2). The combined organic layers were concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0, DCM: methanol = 2: 1). The residue was purified by preparative HPLC (column: Xtimate C18 150*25mm*5μm; mobile phase: [water (10mM NH4HCO3)-ACN]; B%: 20%-50%, 8 minutes). Example 30 was obtained as a white solid, N-[6-cyano-1-[(1-oxo-2H-isoquinolin-5-yl)sulfonyl]indolin-4-yl]acetamide (20 mg, 96.9% purity). LCMS (ESI+) m / z: 409.1.

[0358] 1 H NMR (400MHz, DMSO-d6): δ11.67(br,1H),9.58(s,1H),8.49(d,J=8Hz,1H),8.33(d,J=8Hz,1H),7.83(s,1H),7.61( t,J=8Hz,1H),7.36~7.28(m,2H),7.07(d,J=7.2Hz,1H),4.04(t,J=8.4Hz,2H),2.97(t,J=8.4Hz,2H),2.03(s,1H).

[0359] Example 31: A mixture of N-[6-cyano-1-[(1-oxo-2H-isoquinolin-5-yl)sulfonyl]indolin-4-yl]acetamide (40 mg, 97.94 μmol, 1 eq) in concentrated hydrochloric acid (6 mL) and EtOH (6 mL) was heated to 50°C and stirred for 4.5 h. The reaction mixture was added to an aqueous Na2CO3 solution until pH>7, then diluted with H2O (20 mL) and extracted with DCM (50 mL*2). The combined organic layers were concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (chromatographic column: Welch Ultimate AQ-C18 150*30mm*5μm; mobile phase: [water (0.1% TFA)-ACN]; B%: 25%-55%, 12 minutes). Example 31 was obtained as a white solid as 4-amino-1-[(1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrile (14 mg, 98.04% purity, TFA salt). LCMS (ESI+) m / z: 367.1.

[0360] 1 H NMR (400MHz, DMSO-d6): δ11.69(d,J=5.6Hz,1H),8.51(d,J=7.6Hz,1H),8.27(d,J=7.6Hz,1H),7.63(t,J=7.8Hz,1H),7.37(t ,J=6.8Hz,1H),7.13(d,J=7.6Hz,1H),6.78(s,1H),6.61(s,1H),5.65(br,2H),4.05(t,J=8.4Hz,2H),2.77(t,J=8.4Hz,2H).

[0361] Examples 32, 33 and 34

[0362]

[0363] N-(6-cyano-1-((4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indolin-4-yl)acetamide

[0364]

[0365] 4-Amino-1-((4-methyl-1-oxy-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

[0366]

[0367] 4-(Dimethylamino)-1-((4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

[0368]

[0369] Compound 32-2: Three reaction mixtures were prepared in parallel as follows: KNO3 (53.45 g, 528.71 mmol, 1.1 eq) was added to a solution of 4-bromoisoquinoline (100 g, 480.64 mmol, 1 eq) in H2SO4 (250 mL) at 0°C. The mixture was stirred at 25°C for 12 h under N2 atmosphere. The pH was adjusted to 8 at 0°C by adding to ice water (1 L), and then aqueous NaOH solution (800 mL) was added to the mixture. The mixture was filtered and the filter cake was concentrated under reduced pressure to obtain a residue. The residue was triturated with petroleum ether: EtOAc = 1: 1.5 (2.5 L) for 12 hours at 25°C. Filtered, the solid was triturated with H2O (1.5 L) at 25°C for 5 hours. Compound 32-2 was obtained as a yellow solid, which was 4-bromo-5-nitro-isoquinoline (200 g, 790.35 mmol, 54.81% yield). LCMS (ESI+) m / z: 252.9.

[0370] Compound 32-3: Two reaction mixtures were prepared in parallel as follows: a mixture of 4-bromo-5-nitro-isoquinoline (50 g, 197.59 mmol, 1 eq), Cs2CO3 (128.76 g, 395.18 mmol, 2 eq), Pd(DPPF)Cl2 (14.46 grams, 19.76 mmol, 0.1 eq) and MEB(OH)2 (23.66 g, 395.18 mmol, 2 eq) in dioxane (400 mL) and H2O (40 mL) was degassed and purged 3 times with N2, and then the mixture was stirred at 110°C under N2 atmosphere for 12 h. Two batches of the mixture with the same scale (50 g) were run in parallel and post-processed / purified together. The reaction mixture was concentrated under reduced pressure to remove dioxane. The residue was diluted with H2O (1.5 L) and extracted with EtOAc (2 L * 3). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 1:1). Compound 32-3 was obtained as a yellow solid, which is 4-methyl-5-nitro-isoquinoline (40 g, 212.56 mmol, 53.79% yield).

[0371] Compound 32-4: To a solution of 4-methyl-5-nitroisoquinoline (60 g, 318.84 mmol, 1 eq) in DCM (800 mL) was added m-CPBA (151.31 g, 701.44 mmol, 80% purity, 2.2 eq) at 0°C. The mixture was stirred at 25°C for 12 h. The reaction mixture was quenched by the addition of an aqueous solution of Na2SO3 (900 mL). The reaction mixture was diluted with H2O (1 L) and extracted with DCM (2000 mL*4). The combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was triturated with EtOAc (2000 mL) at 25°C for 2 hours, and the solid was dried under reduced pressure. Compound 32-4 was obtained as a yellow solid as 4-methyl-nitro-2-diamino-isoquinolin-2-ium (40 g, 195.90 mmol, 61.44% yield).

[0372] Compound 32-5: POCl3 (40.55 g, 264.47 mmol, 24.58 mL, 1.2 eq) and DMF (8.05 g, 110.20 mmol, 8.48 mL 0.5 eq) were added to a solution of 4-methyl-5-nitro-2-oxido-isoquinolin-2-ium (45 g, 220.39 mmol, 1 eq) in DCM (500 mL) at 0 ° C. The mixture was stirred at 20 ° C for 12 h under N2 atmosphere. The reaction mixture was concentrated under reduced pressure to remove DCM. The residue was quenched with H2O (300 mL) and then filtered. The filter cake was dried under reduced pressure to obtain a residue, which can be used without further purification. The crude product compound 32-5 was obtained as a yellow solid, which was 1-chloro-4-methyl-5-nitro-isoquinoline (27 g, crude product). LCMS (ES1+) m / z: 222.9.

[0373] Compound 32-6: A mixture of 1-chloro-4-methyl-5-nitro-isoquinoline (27 g, 121.28 mmol, 1 eq), NaOMe (13.10 g, 242.56 mmol, 2 eq) in methanol (500 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 80 ° C for 5 h under N2 atmosphere. The reaction mixture was concentrated under reduced pressure to remove methanol. The residue was diluted with H2O (500 mL) and extracted with EtOAc (600 mL*4). The combined organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 5:1). Compound 32-6 was obtained as a yellow solid, which was 1-methoxy-4-methyl-5-nitro-isoquinoline (15 g, 68.74 mmol, 56.68% yield).

[0374] Compound 32-7: To a solution of 1-methoxy-4-methyl-5-nitro-isoquinoline (15 g, 68.74 mmol, 1 eq) in methanol (210 mL) and H2O (70 mL) was added Zn (22.48 g, 343.71 mmol, 5 eq) and NH4Cl (18.39 g, 343.71 mmol, 5 eq) at 0°C. The mixture was stirred at 25°C for 12 h. The mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 1:1). Compound 32-7 was obtained as 1-methoxy-4-methyl-isoquinoline-5-amine (10 g, 53.13 mmol, 77.29% yield) as a brown solid. LCMS (ESI+) m / z: 189.0.

[0375] Compound 32-8: To a solution of 1-methoxy-4-methyl-isoquinolin-5-amine (2 g, 10.63 mmol, 1 eq) in HCl (80 mL) was added a solution of NaNO2 (903.12 mg, 10.63 mmol, 1 eq) in H2O (5 mL) at -5 °C. The mixture was stirred at -5 °C for 1 h and transferred in batches to a solution of AcOH (50 mL) and CuCl (262.98 mg, 2.66 mmol, 63.52 μL, 0.25 eq) in H2O (5 mL), which was saturated with SO2. The mixture was stirred at 20 °C for 2 h. At 0 °C, the reaction mixture was quenched by adding ice water (100 mL), and the mixture was then extracted with DCM (200 mL*3). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue that was used without further purification. Compound 32-8, 1-methoxy-4-methyl-isoquinoline-5-sulfonyl chloride (1.5 g, crude) was obtained as a yellow solid. LCMS (ES1+) m / z: 271.9.

[0376] Compound 32-9: At -5 ° C, to a mixture of N-(6-cyanoindolyl-4-yl)acetamide (888.67 mg, 4.42 mmol, 1.2 eq) and pyridine (1.05 g, 13.25 mmol, 1.07 mL, 3.6 eq) was added dropwise a solution of 1-methoxy-4-methylisoquinoline-5-sulfonyl chloride (780 mg, 2.87 mmol, 0.78 eq) in DCM (10 mL). The mixture was degassed and purged with N2 3 times, and then the mixture was stirred at 15 ° C for 12 h under N2 atmosphere. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue. The crude compound 32-9 was obtained as a brown solid, which was N-(6-cyano-1-((1-methoxy-4-methylisoquinoline-5-yl)sulfonyl)indolyl-4-yl)acetamide (1.2 g, crude product), which was used in the next step without further purification.

[0377] Example 32: A mixture of N-(6-cyano-1-((1-methoxy-4-methylisoquinolin-5-yl)sulfonyl)indolin-4-yl)acetamide (600 mg, 1.37 mmol, 1 eq) and TMSI (1.10 g, 5.50 mmol, 748.43 μL, 4 eq) in ACN (10 mL) was stirred at 15 ° C. for 1 h under N2 atmosphere. At 0 ° C., the reaction mixture was quenched by adding H2O (30 mL), and then extracted with EtOAc (30 mL*2). The combined organic layers were washed with brine (60 mL×1), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative TLC (SiO2, DCM: methanol = 10: 1), and then repurified by preparative HPLC (column: Waters Xbridge BEH C18 100*25mm*5μm; mobile phase: [water (10mM NH4HCO3)-ACN]; B%: 15%-50%, 8 minutes). Example 32 was obtained as a white solid, N-(6-cyano-1-((4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indolin-4-yl)acetamide (4 mg, 9.46umol, 99.9% purity). LCMS (ESI+) m / z: 423.1.

[0378] 1H NMR (400 MHz, DMSO-d6): δ11.69 (br, 1H), 9.74 (s, 1H), 8.59 (d, J = 4 Hz, 1H), 7.98-7.85 (m, 2H), 7.52 (t, J = 7.6 Hz, 1H), 7.24 (s, 1H), 6.99 (s, 1H), 4.22 (t, J = 8.4 Hz, 2H), ~3.31 (2H, mixed with solvent peak), 2.59 (s, 3H), 2.14 (s, 3H).

[0379] Example 33: A solution of N-(6-cyano-1-((4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indolin-4-yl)acetamide (30 mg, 71.01 μmol, 1 eq) in concentrated hydrochloric acid (5 mL) and EtOH (5 mL) was stirred at 50°C for 5 h. At 0°C, saturated Na2CO3 solution was added to the reaction mixture until pH = 8, and then extracted with EtOAc (60 mL*5). The combined organic layers were washed with brine (200 mL*1), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (chromatographic column: Waters Xbridge BEH C18 100*25mm*5μm; mobile phase: [water (10mM NH4HCO3)-ACN]; B%: 20%-50%, 8 minutes). Example 33 was obtained as 4-amino-1-(((4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile (3.8 mg, 9.87 umol, 13.89% yield, 98.77% purity) as a white solid. LCMS (ESI+) m / z: 381.0.

[0380] 1 H NMR (400MHz, DMSO-d6): δ11.66(br,1H),8.57(d,J=7.6Hz,1H),7.81(d,J=7.6Hz,1H),7.53(t,J=7.8Hz,1H),7 .23(s,1H),6.64(s,1H),6.48(s,1H),5.79(s,2H),4.17(t,J=8.4Hz,2H),3.07(t,J=8.4Hz,2H),2.59(s,3H).

[0381] Example 34: To a solution of 4-amino-1-((4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile (20 mg, 52.57 μmol, 1 eq) in DMF (2 mL) was added pyridine (12.48 mg, 157.72 μmol, 12.73 μL, 3 eq) and MeI (74.62 mg, 525.74 μmol, 32.73 μL, 10 eq). The mixture was then stirred at 25 ° C for 12 h. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (chromatographic column: Waters Xbridge BEH C18100*25mm*5μm; mobile phase: [water (10mM NH4HCO3)-ACN]; B%: 30%-60%, 8 minutes). 4-(Dimethylamino)-1-((4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile (2.1 mg, 5.09 umol, 9.68% yield, 99% purity) was obtained as a white solid. LCMS (ESI+) m / z: 409.0.

[0382] 1 H NMR (400 MHz, DMSO-d6): δ11.68 (br, 1H), 8.59 (d, J=7.6 Hz 1H), 7.86 (d, J=7.6 Hz, 1H), 7.53 (t, J=8 Hz, 1H), 7.24 (s, 1H), 6.93 (s, 1H), 6.78 (s, 1H), 4.17 (t, J=8.4 Hz, 2H), ~3.05 (2H, mixed with solvent peak), 2.87 (s, 6H), 2.61 (s, 3H).

[0383] Examples 35, 36 and 37

[0384]

[0385] N-(1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-6-cyanoindolin-4-yl)acetamide

[0386]

[0387] 4-Amino-1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoleline-6-carbonitrile

[0388]

[0389] 1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-4-(dimethylamino)indoline-6-carbonitrile

[0390]

[0391] Compound 35-2: At -5°C, a solution of 4-chloroisoquinoline (40 g, 244.49 mmol, 1 eq) in H2SO4 (110 mL) was slowly added dropwise to a solution of KNO3 (32.13 g, 317.84 mmol, 1.3 eq) in H2SO4 (140 mL). The mixture was stirred at 0°C for 1 h and then at 15°C for 12 h. The mixture was poured into ice water (3 L), and solid Na2CO3 was then added to the mixture until pH = 8. A yellow solid was precipitated, filtered, and the filter cake was washed with H2O (2000 mL*3), and the filter cake was concentrated. The filter cake was dissolved in EtOAc, the EtOAc was filtered, and the filter cake was concentrated under reduced pressure to obtain a residue. The crude compound 35-2 was obtained as a yellow solid as 4-chloro-5-nitroisoquinoline (43 g, 206.14 mmol, 84.31% yield).

[0392] Compound 35-3: At 0°C, m-CPBA (88.93 g, 412.27 mmol, 80% purity, 2 eq) was added to a solution of 4-chloro-5-nitroisoquinoline (43 g, 206.14 mmol, 1 eq) in CHCl3 (500 mL) in multiple batches. The mixture was stirred at 15°C for 12 h. Saturated NaHCO3 (1 L) was added to the mixture at 0°C, and the mixture was extracted with CHCl3 / methanol = 10 / 1 (500 mL*4), and then the organic layer was washed with 15% Na2S2SO4 (500 mL*4) aqueous solution. The combined organic layers were dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 1 to 0 / 1). Compound 35-3 was obtained as a yellow solid as 4-chloro-5-nitroisoquinoline 2-oxide (36 g, 160.28 mmol, 77.76% yield).

[0393] Compound 35-4: POCl3 (49.15 g, 320.57 mmol, 29.79 mL, 2 eq) was slowly added to a solution of 4-chloro-5-nitroisoquinoline 2-oxide (36 g, 160.28 mmol, 1 eq) in DCM (500 mL) at 0 ° C. The mixture was stirred at 60 ° C for 12 h under N2 atmosphere. The mixture was added to water (1000 mL) and extracted with DCM (500 mL*3). The organic layer was washed with saturated NaHCO3 (800 mL*3) and brine (1500 mL), then dried over Na2SO4, filtered and concentrated. Crude compound 35-4 was obtained as a brown solid, which was 1,4-dichloro-5-nitroisoquinoline (26 g, 106.98 mmol, 66.74% yield).

[0394] Compound 35-5: NaOMe (24.45 g, 113.15 mmol, 25% purity, 1.1 eq) was added to a solution of 1,4-dichloro-5-nitroisoquinoline (25 g, 102.86 mmol, 1 eq) in methanol (150 mL). The mixture was stirred at 80 ° C. for 1 h under N2 atmosphere, filtered and the filter cake was concentrated under reduced pressure to obtain compound 36-5 as a brown solid, which is 4-chloro-1-methoxy-5-nitroisoquinoline (16 g, 67.05 mmol, 65.19% yield).

[0395] Compound 35-6: At 0°C, Zn (43.57 g, 666.31 mmol, 10 eq) was slowly added to a solution of 4-chloro-1-methoxy-5-nitroisoquinoline (15.9 g, 66.63 mmol, 1 eq) and NH4Cl (35.64 g, 666.31 mmol, 10 eq) in methanol (200 mL) and H2O (40 mL). The mixture was stirred at 15°C for 2 h, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was added to H2O (50 mL), and the mixture was extracted with EtOAc (500 mL*4). The combined organic layer was washed with brine (2000 mL*1), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was washed with petroleum ether / ethyl acetate = 30:1 (930 mL), filtered, and the filter cake was dried under reduced pressure. The crude compound 36-6 was obtained as 4-chloro-1-methoxyisoquinolin-5-amine (11.2 g, 53.68 mmol, 80.56% yield) as a white solid.

[0396] Preparation of solution A: To a solution of 4-chloro-1-methoxyisoquinolin-5-amine (0.2 g, 958.57 umol, 1 eq) in HCl (8 mL) was added a solution of NaNO2 (99.21 mg, 1.44 mmol, 1.5 eq) in H2O (1 mL) at -5°C. The mixture was stirred at -5°C for 1 h.

[0397] Preparation of Solution B: SO 2 was bubbled at 15 psi for 30 min into a solution of CuCl (23.72 mg, 239.64 mmol, 5.73 uL, 0.25 eq) in AcOH (5 mL) and H 2 O (1 mL) at 0° C.

[0398] Compound 35-7: Solution A was transferred to solution B, and the mixture was stirred at 15 ° C for 2 h. At 0 ° C, the reaction mixture was quenched by adding to ice water (20 mL), and then extracted with DCM (30 mL * 3). The combined organic layer was washed with brine (100 mL * 2), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. Compound 35-7 was obtained as a yellow solid, which was 4-chloro-1-methoxyisoquinoline-5-sulfonyl chloride (300 mg, crude product).

[0399] Compound 35-9: At -5 ° C, to a mixture of N-(6-cyanoindolyl-4-yl)acetamide (compound 35-8; 500 mg, 2.48 mmol, 1.3 eq) and pyridine (453.57 mg, 5.73 mmol, 462.83 μL, 3 eq) was added dropwise a solution of 4-chloro-1-methoxyisoquinoline-5-sulfonyl chloride (1.27 g, 1.91 mmol, 1 eq) in DCM (5 mL). The mixture was degassed and purged with N2 3 times, and the mixture was stirred at 15 ° C for 12 h under N2 atmosphere. The reaction mixture was concentrated under reduced pressure to obtain a residue. Crude compound 35-9 was obtained as a red solid, which was N-(1-((4-chloro-1-methoxyisoquinoline-5-yl)sulfonyl)-6-cyanoindolyl-4-yl)acetamide (1.5 g, crude product). It can be used in the next step without further purification.

[0400] Example 35: A mixture of N-(1-((4-chloro-1-methoxyisoquinolin-5-yl)sulfonyl)-6-cyanoindolin-4-yl)acetamide (100 mg, 218.87 umol, 1 eq) and TMSI (218.96 mg, 1.09 mmol, 148.96 uL, 5 eq) in ACN (10 mL) was stirred at 80 ° C for 2 h under N2 atmosphere. The reaction mixture was quenched by adding to H2O (20 mL) at 0 ° C, and then extracted with EtOAc (40 mL*2). The combined organic layers were washed with brine (80 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative TLC (SiO2, DCM: methanol = 10: 1). The residue was then repurified by preparative HPLC (column: Waters Xbridge BEH C18 100*25mm*5μm; mobile phase: [water (10mM NH4HCO3)-ACN]; B%: 20%-50%, 8 minutes). Example 35 was obtained as a white solid, N-(1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-6-cyanoindolin-4-yl)acetamide (1.9 mg, 4.08umol, 1.86% yield, 95.1% purity). LCMS (ESI+) m / z: 443.0.

[0401] 1 H NMR (400 MHz, DMSO-d6): δ9.73 (s, 1H), 8.57 (d, J = 7.6 Hz, 1H), 8.01 (d, J = 8 Hz, 1H), 7.91 (s, 1H), 7.66 (s, 1H), 7.63 (t, J = 7.6 Hz, 1H), 6.98 (s, 1H), 4.19 (t, J = 8.4 Hz, 2H), ~3.20 (2H, mixed with solvent peak), 2.14 (s, 3H).

[0402] Example 36: A solution of N-(1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-6-cyanoindolin-4-yl)acetamide (16 mg, 36.13 umol, 1 eq) in concentrated hydrochloric acid (3 mL) and EtOH (3 mL) was stirred at 50°C for 5 h. To the mixture was added saturated Na2CO3 solution at 0°C until pH = 8, and the mixture was extracted with EtOAc (40 mL*5). The combined organic layers were washed with brine (200 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (chromatographic column: Waters Xbridge BEH C18 100*25mm*5μm; mobile phase: [water (10mMNH4HCO3)-ACN]; B%: 20%-40%, 8 minutes). Example 36, 4-amino-1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile (1.6 mg, 97.6% purity) was obtained as a white solid. LCMS (ESI+) m / z: 400.9.

[0403] 1 H NMR (400MHz, DMSO-d6): δ12.04(br,1H),8.56(d,J=8Hz,1H),7.92(d,J=7.6Hz,1H),7.65~7.6 1(m,2H),6.63(s,1H),6.45(s,1H),5.77(s,2H),4.15(t,J=8.4Hz,2H),3.05(t,J=8.4Hz,2H).

[0404] Example 37: To a solution of 4-amino-1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile (20 mg, 49.90 μmol, 1 eq) in DMF (2.5 mL) was added pyridine (39.47 mg, 498.95 mmol, 40.27 μL, 10 eq) and MeI (141.64 mg, 997.91 mmol, 62.12 μL, 20 eq). The mixture was stirred at 25 ° C for 12 h. The mixture was concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (chromatographic column: Waters Xbridge BEHC18100*25 mm*5 μm; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 30%-60%, 8 minutes). Example 37 was obtained as 1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-4-(dimethylamino)indoline-6-carbonitrile (3.1 mg, 7.08 μmol, 14.20% yield, 98% purity) as a white solid. LCMS (ESI+) m / z: 429.1.

[0405] 1 HNMR (400 MHz, DMSO-d6): δ12.05 (br, 1H), 8.55 (d, J=8 Hz, 1H), 7.94 (d, J=8 Hz, 1H), 7.65 (s, 1H), 7.63 (t, J=8 Hz, 1H), 6.92 (s, 1H), 6.74 (s, 1H), 4.13 (t, J=8.4 Hz, 2H), ~3.33 (2H, mixed with solvent peak), 2.85 (s, 6H).

[0406] Embodiment 38

[0407]

[0408] 1-((1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine-6-carbonitrile

[0409]

[0410] Example 38: At -5 ° C, to a mixture of 2,3-dihydro-1H-pyrrolo[3,2-b]pyridine-6-carbonitrile (21.45 mg, 147.74 μmol, 1.2 eq) and pyridine (29.22 mg, 369.36 μmol, 29.81 μL, 3 eq) was added dropwise a solution of 1-oxo-2H-isoquinoline-5-sulfonyl chloride (30 mg, 123.12 μmol, 1 eq) in DCM (3 mL). The mixture was degassed and purged with N2 3 times, and the mixture was stirred at 25 ° C for 12 h under N2 atmosphere. The reaction mixture was concentrated under reduced pressure to give a residue. The crude product was absorbed in methanol (2 mL) at 25 ° C for 10 minutes, filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (chromatographic column: Welch Ultimate AQ-C18 150*30mm*5pM; mobile phase: [watcr(0.1%TFA)-ACN]; B%: 35%-65%, 12 minutes). Example 38 was obtained as a white solid, 1-[(1-oxo-2H-isoquinolin-5-yl)sulfonyl]-2,3-dihydropyrrolo[3,2-b]pyridine-6-carbonitrile (15 mg, 41.51 umol, 33.71% yield, 97.5% purity). LCMS (ESI+) m / z: 353.0.

[0411] 1 H NMR (400MHz, DMSO-d6): δ11.71(br,1H),8.59~8.52(m,2H),8.46(d,J=7.6Hz,1H),7.90(d,J=2Hz,1H),7.64 (t, J=7.6Hz, 1H), 7.39 (t, J=7.6Hz, 1H), 7.11 (d, J=7.6Hz, 1H), 4.08 (t, J=8.4Hz, 2H), 3.22 (t, J=8.8Hz, 2H).

[0412] Embodiment 39

[0413]

[0414] 1-((4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine-6-carbonitrile

[0415]

[0416] Compound 39-2: At -5 ° C, to a mixture of 2,3-dihydro-1H-pyrrolo[3,2-b]pyridine-6-carbonitrile (19.23 mg, 132.49 μmol, 1.2 eq) and pyridine (26.20 mg, 331.22 μmol, 26.73 μL, 3 eq) was added dropwise a solution of 1-methoxy-4-methyl-isoquinoline-5-sulfonyl chloride (30 mg, 110.41 μmol, 1 eq) in DCM (2 mL). The mixture was degassed and purged with N2 3 times, and the mixture was stirred at 25 ° C for 12 h under N2 atmosphere. The reaction mixture was concentrated under reduced pressure to remove DCM (2 mL). The residue was purified by preparative TLC (SiO2, petroleum ether / ethyl acetate = 1 / 1). Compound 39-2, 1-[(1-methoxy-4-methyl-5-isoquinolyl)sulfonyl]-2,3-dihydropyrrolo[3,2-b]pyridine-6-carbonitrile (15 mg, 39.43 umol, 35.71% yield) was obtained as a white solid.

[0417] Example 39: A mixture of 1-[(1-methoxy-4-methyl-5-isoquinolyl)sulfonyl]-2,3-dihydropyrrolo[3,2-b]pyridine-6-carbonitrile (15 mg, 39.43 μmol, 1 eq) and TMSI (31.56 mg, 157.72 μmol, 21.47 μL, 4 eq) in ACN (2 mL) was stirred at 25 °C for 1 h under N2 atmosphere. The reaction mixture was quenched by adding H2O (5 mL) at 0 °C, and then extracted with EtOAc (5 mL*2). The combined organic layers were washed with brine (5 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (chromatographic column: Waters Xbridge Prep OBD C18150*40mm*10 μm; mobile phase: [water (0.04% NH 3. H20+10 mM NH4HCO3)-ACN]; B%: 25%-55%, 8 minutes). Example 39 was obtained as a white solid, 1-[(4-methyl-1-oxo-2H-isoquinolin-5-yl)sulfonyl]-2,3-dihydropyrrolo[3,2-b]pyridine-6-carbonitrile (3.4 mg, 9.28 umol, 23.53% yield, 100% purity). LCMS (ESI+) m / z: 367.0.

[0418] 1H NMR (400MHz, DMSO-d6): δ11.72(br,1H),8.62~8.57(m,2H),8.02(d,J=2.4Hz,1H),7.5 9~7.51(m,2H),7.25(s,1H),4.23(t,J=8.8Hz,2H),3.49(t,J=8.8Hz,2H),2.59(s,3H).

[0419] Embodiment 40

[0420]

[0421] 1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine-6-carbonitrile

[0422]

[0423] Compound 40-2: At -5 ° C, to a mixture of 2,3-dihydro-1H-pyrrolo[3,2-b]pyridine-6-carbonitrile (17.89 mg, 123.23 umol, 1.2 eq) and pyridine (24.37 mg, 308.07 μmol, 24.87 μL, 3 eq) was added dropwise a solution of 4-chloro-1-methoxy-isoquinoline-5-sulfonyl chloride (30 mg, 102.69 μmol, 1 eq) in DCM (2 mL). The mixture was degassed and purged with N2 3 times, and the mixture was stirred at 25 ° C for 12 h under N2 atmosphere. LC-MS showed that ~50% of the desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove DCM (2 mL). The residue was purified by preparative TLC (SiO2, PE: EA = 1: 1).

[0424] Compound 40-2, 1-[(4-chloro-1-methoxy-5-isoquinolyl)sulfonyl]-2,3-dihydropyrrolo[3,2-b]pyridine-6-carbonitrile (30 mg, 74.84 umol, 72.88% yield), was obtained as a white solid.

[0425] Example 40: A mixture of 1-[(4-chloro-1-methoxy-5-isoquinolyl)sulfonyl]-2,3-dihydropyrrolo[3,2-b]pyridine-6-carbonitrile (30 mg, 74.84 umol, 1 eq) and TMSI (59.90 mg, 299.37 umol, 40.75 uL, 4 eq) in ACN (2 mL) was stirred at 25 ° C for 1 h under N2 atmosphere. The reaction mixture was quenched by adding H2O (4 mL) at 0 ° C, and then extracted with EtOAc (4 mL*2). The combined organic layers were washed with brine (8 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (chromatographic column: Welch Ultimate AQ-C18 150*30mm*5μm; mobile phase: [water (0.1% TFA)-ACN]; B%: 15%-45%, 12 minutes). Example 40 was obtained as a yellow solid, 1-[(4-chloro-1-oxo-2H-isoquinolin-5-yl)sulfonyl]-2,3-dihydropyrrolo[3,2-b]pyridine-6-carbonitrile (7 mg, 17.55 umol, 23.45% yield, 97% purity). LCMS (ESI+) m / z: 386.9.

[0426] Indoline reagent

[0427] Indoline 2

[0428]

[0429] 6-Chloroindoline

[0430]

[0431] Indoline 2: At 0 ° C, NaBH3CN (6.84 g, 108 mmol) was added to a mixture of 6-chloro-1H-indole (5 g, 32.98 mmol) in AcOH (80 mL), and the mixture was then stirred at 25 ° C for 12 h under N2 atmosphere. At 0 ° C, the reaction mixture was quenched by adding ice water (20 mL), and then a saturated aqueous NaOH solution was added to the mixture to adjust the pH to ~ 8. The mixture was extracted with EtOAc (250 mL × 2). The combined organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 20: 1 to 5: 1). The title compound was obtained as a green oil, which was 6-chloroindoline (3 g, 19.5 mmol, 59.2% yield).

[0432] Indoline 4

[0433]

[0434] Indoline-6-carbonitrile

[0435]

[0436] Indoline 4: At 0 ° C, NaBH3CN (7.29 g, 116.0 mmol) was added to a mixture of 1H-indole-6-carbonitrile (5 g, 35.1 mmol) in AcOH (80 mL), and the mixture was then stirred at 25 ° C for 12 h under N2 atmosphere. At 0 ° C, the reaction mixture was quenched by adding ice water (20 mL). Saturated NaOH aqueous solution was added to the mixture to adjust the pH to ~~8. The mixture was extracted with EtOAc (300 mL×2). The combined organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate=10: 1 to 5: 1). The residue was purified by preparative HPLC (column: Xtimate C18 10μ x 250mm x 50mm; mobile phase: [A, water (10mM NH4HCO3)-B, ACN]; B%: 20%-40%, 25 minutes). The title compound was obtained as a white solid, indoline-6-carbonitrile (1.9 g, 13.1 mmol, 37.4% yield).

[0437] Indoline 6

[0438]

[0439] 1-(1H-Indol-4-yl)ethan-1-one

[0440] Indoline 7

[0441]

[0442] 1-(1H-Indol-4-yl)ethan-1-ol

[0443] Indoline 8

[0444]

[0445] 1-(Indolin-4-yl)ethan-1-ol

[0446]

[0447] Indoline 6: MeLi (1.8M, 41.9mL) was added to a solution of 1H-indole-4-carbonitrile (5g, 35.1mmol) in THF (20mL). The mixture was stirred at 80°C for 8h under N2 atmosphere. The reaction mixture was quenched by adding H2O (100mL) at 0°C. The reaction mixture was extracted with DCM (400mL×2). The combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate=20:1 to 1:1). The title compound was obtained as a yellow solid, 1-(1H-indol-4-yl)ethanone (2.5g, 15.7mmol, 44.6% yield).

[0448] Indoline 7: At 0°C, NaBH4 (2.28 g, 60.3 mmol) was added to a solution of 1-(1H-indol-4-yl)ethanone (2.4 g, 15.0 mmol) in THF (20 mL). The mixture was stirred at 25°C for 12 h under N2 atmosphere. The reaction mixture was quenched by adding H2O (100 mL) at 0°C. The reaction mixture was extracted with DCM (250 mL×2). The combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate=1:1). 1-(1H-indol-4-yl)ethanol (1 g, 6.20 mmol, 41.1% yield) was obtained as a white solid.

[0449] Indoline 8: At 0 ° C, NaBH3CN (1.29 g, 20.4 mmol) was added to a mixture of 1-(1H-indol-4-yl)ethanol (1 g, 6.20 mmol) in AcOH (10 mL), and the mixture was stirred at 25 ° C for 5 h under N2 atmosphere. At 0 ° C, the reaction mixture was quenched by adding ice water (15 mL), and a saturated aqueous NaOH solution was added to the mixture to adjust the pH to ~ 8. The reaction mixture was extracted with DCM (100 mL × 2). The combined organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10: 1 to 1: 1). 1-indol-4-ylethanol (250 mg, 1.53 mmol, 24.6% yield) was obtained as a white solid.

[0450] Indoline 10

[0451]

[0452] 4,6-Dichloroindoline

[0453]

[0454] Indoline 10: NaBH3CN (557 mg, 8.87 mmol) was added to a mixture of 4,6-dichloro-1H-indole (0.5 g, 2.69 mmol) in AcOH (5 mL) at 0 ° C, and the mixture was stirred at 25 ° C for 12 h under N2 atmosphere. At 0 ° C, the reaction mixture was quenched by adding ice water (20 mL), and a saturated aqueous NaOH solution was added to the mixture to adjust the pH to ~8. The reaction mixture was extracted with DCM (100 mL×2). The combined organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate=30:1 to 5:1). 4,6-dichloroindoline (0.1 g, 531.7 μmol, 19.7% yield) was obtained as a yellow oil.

[0455] Indoline 12

[0456]

[0457] 4-Cyclopropyl-1H-indole-6-carbonitrile

[0458] Indoline 13

[0459]

[0460] 4-Cyclopropylindoline-6-carbonitrile

[0461]

[0462] Indoline 12: A mixture of 4-bromo-1H-indole-6-carbonitrile (0.1 g, 452.3 μmol), cyclopropylboronic acid (77.7 mg, 904.7 μmol), Cs2CO3 (294.7 mg, 904.7 μmol), Pd(dppf)Cl2 (33.1 mg, 45.2 μmol) in dioxane (5 mL) and H2O (0.5 mL) was degassed and purged with N2 three times. The mixture was then stirred at 110°C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (20 mL) and extracted with EtOAc (80 mL×2). The combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate=20:1 to 5:1). 4-Cyclopropyl-1H-indole-6-carbonitrile (50 mg, 274.3 μmol, 60.6% yield) was obtained as a yellow solid.

[0463] Indoline 13: Sodium cyanoborohydride (56.90 mg, 905.4 μmol) was added to a solution of 4-cyclopropyl-1H-indole-6-carbonitrile (50 mg, 274.3 μmol) in AcOH (3 mL) at 0 ° C. The mixture was stirred at 25 ° C for 12 h. The reaction mixture was quenched by adding ice water (5 mL) at 0 ° C, and a saturated aqueous NaOH solution was added to the mixture to adjust the pH to ~8. The reaction mixture was extracted with DCM (50 mL×2). The combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative TLC (SiO2, petroleum ether / ethyl acetate=5:1).

[0464] The compound, 4-cyclopropylindoline-6-carbonitrile (20 mg, 108.6 μmol, 39.5% yield) was obtained as a yellow solid.

[0465] Indoline 15

[0466]

[0467] 4-Methyl-1H-indole-6-carbonitrile

[0468] Indoline 16

[0469]

[0470] 4-Methylindoline-6-carbonitrile

[0471]

[0472] Indoline 15: To a mixture of 4-bromo-1H-indole-6-carbonitrile (1 g, 4.52 mmol) in dioxane (15 mL) and H2O (1.5 mL), methylboric acid (541.5 mg, 9.05 mmol), Cs2CO3 (2.95 g, 9.05 mmol) and Pd(dppf)Cl2 (331.0 mg, 452.3 μmol) were added in batches. The mixture was heated to 110°C and stirred for 12 h. The reaction mixture was diluted with H2O (300 mL) and extracted with EtOAc (150 mL×2). The combined organic layers were washed with brine (300 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate=1:0 to 0:1). 4-Methyl-1H-indole-6-carbonitrile (600 mg, 3.34 mmol, 73.8% yield) was obtained as a light yellow solid.

[0473] Indoline 16: NaBH3CN (1.45 g, 23.0 mmol) was added to a solution of 4-methyl-1H-indole-6-carbonitrile (600 mg, 3.84 mmol) in HOAc (15 mL). The mixture was stirred at 25 °C for 12 h. Saturated aqueous Na2CO3 solution was added to the reaction mixture until the pH value was about 8. The mixture was diluted with H2O (300 mL) and extracted with EtOAc (150 mL×3). The combined organic layers were washed with brine (300 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate=1:0 to 0:1). 4-Methylindoline-6-carbonitrile (250 mg, 1.20 mmol, 31.2% yield) was obtained as a light yellow solid.

[0474] Indoline 18

[0475]

[0476] 4-(Propan-1-en-2-yl)indoline

[0477]

[0478] Indoline 18: 4-bromoindoline (0.5 g, 2.52 mmol), 2-isopropenyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (509.0 mg, 3.03 mmol), Cs2CO3 (1.23 g, 3.79 mmol), Pd(dppf)Cl2 (184.7 mg, 252.4 μmol) in dioxane (10 mL) and H2O (2 mL) solution was degassed and purged with N2 three times, and then the mixture was stirred at 110 ° C. for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O (20 mL) and extracted with DCM (60 mL×3). The combined organic layer was washed with saturated brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate=1:0 to 10:1). 4-Isopropenylindoline (60 mg, 376.8 μmol, 14.9% yield) was obtained as a yellow solid.

[0479] Indoline 20

[0480]

[0481] 4-Vinylindoline

[0482]

[0483] Indoline 20: To a mixture of 4-bromoindoline (1 g, 5.05 mmol) and 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (933 mg, 6.06 mmol) in dioxane (10 mL) and H2O (2 mL) was added Pd(dppf)Cl2 (369 mg, 504.9 μmol) and Cs2CO3 (2.47 g, 7.57 mmol) at 15 °C and N2. The mixture was heated to 110 °C and stirred for 12 h. The residue was diluted with H2O (120 mL) and extracted with EtOAc (60 mL×2). The combined organic layers were washed with 120 mL of brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1:0 to 10:1). 4-Vinylindoline (400 mg, 2.45 mmol, 48.6% yield) was obtained as a yellow oil.

[0484] Indoline 22

[0485]

[0486] 4-Cyclopropylindoline

[0487]

[0488] Indoline 22: Cyclopropylboronic acid (433.6 mg, 5.05 mmol), cesium carbonate (1.65 g, 5.05 mmol) and cyclopentyl (diphenyl) phosphine were added to a solution of 4-bromoindoline (0.5 g, 2.52 mmol) in dioxane (10 mL) and H2O (1 mL). Dichloropalladium; iron (184.7 mg, 252.4 μmol) were added. The mixture was stirred at 110 ° C for 12 h under N2 atmosphere. The reaction mixture was diluted with H2O15 mL and extracted with EtOAc (40 mL×2). The combined organic layer was washed with 20 mL of saturated brine, dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1:0 to 10:1). 4-Cyclopropylindoline (0.1 g, 628 μmol, 24.8% yield) was obtained as a yellow oil.

[0489] Embodiment 41

[0490] This example illustrates that exemplary compounds of the invention inhibit the enzymatic activity of ROCK1 and / or ROCK2.

[0491] The dose-response curves of the compounds of the invention were determined by a fluorescence-coupled enzyme assay using purified C-terminally truncated human ROCK1 catalytic domain (amino acids 1-477 of accession number NP_005397.1) fused to CST (Carna BioSciences, catalog #01-109) and purified C-terminally truncated human ROCK2 enzyme kinase domain (amino acids 1-553 of accession number NP_004841.2) fused to GST (Carna BioSciences, catalog #01-110). In addition, full-length cGMP-dependent protein kinase 1 (PKG or PRKG1; amino acids 1-686 of accession number NP_006249.1) fused to GST (Carna BioSciences, catalog number #01-142), full-length PKACα (PRKACA; amino acids 1-351 of accession number NP_002721.1) fused to GST (Carna BioSciences, catalog number #01-127), and the C-terminally truncated human AKT1 catalytic domain (amino acids 104-480 of accession number NP_005154.1) fused to GST (Carna Biosciences, catalog number #01-101) were used as controls to measure any unwanted off-target kinase activity.

[0492] Briefly, the compounds of the invention were dissolved in DMSO and a series of 12, three-fold serial dilutions were performed for each compound in 2.5% DMSO. The initial starting concentration of the serial dilutions of each compound was 10 μM or 100 μM. Control samples without compound, ROCK enzyme or ATP were also prepared and processed in parallel with the compound test samples.

[0493] A 4 μL aliquot of each serial dilution of the test compound diluted with 2.5% DMSO in H2O was added to wells containing 4 μL of 10 nM purified N-truncated ROCK1 (final concentration 4 nM), 10 nM purified N-truncated ROCK2 enzyme (final concentration 4 nM), 1.25 nM AKT1 (final concentration 0.5 nM), 25 pMPKACA (final concentration 10 pM), or 20 nM PKG (final concentration 8 nM) in reaction buffer (containing 50 mM Hepes, pH 7.5, 1 mM EGTA, 10 mM MgCl2, 2 mM DTT, and 0.01% Tween 20 (final concentration)). To start the enzymatic reaction, 2 μL of solution was added to each well to provide a final concentration of 50 nM Ulight-CREBTide peptide substrate (Perkin Elmer, catalog number TRF0107) and 4 pM ATP for ROCK1 and AKT1 assays, 8 pM ATP for ROCK2 assays and 30 pM ATP for PKG assays, or 1 pM ATP for PKACA assays. The reaction mixture was incubated at room temperature for one hour. One hour later, the reaction was terminated by adding 10 μL of europium-labeled phosphorylated CREB (Ser133) antibody (Perkin Elmer, catalog number TRF0200) in LANCE buffer supplemented with 20 mM EDTA. The terminated reaction mixture was incubated at room temperature for 90 minutes. The ratio of TR-FRET 665 / 620 was determined using a Clariostar monochromator plate reader (BMG Laboratory Technology) according to the manufacturer's instructions. The IC of each compound was determined from each dose response curve using GraphPad Prism 7 software with Sigmodial dose response. 50 Table 1 shows the results for exemplary compounds of formula (I) and formula (II). Key: A is IC 50 ≤100nM: Β is 100nM <IC 50 <1000nM; C is 1,000 <IC 50 ≤10,000nM; D is IC 50 >10,000nM

[0494] Table 1

[0495] Inhibition of ROCK activity by exemplary compounds of formula (I)

[0496]

[0497]

[0498] Although the present invention has been described in conjunction with specific embodiments of the present invention, it will be appreciated that the present invention is capable of further modifications, and this application is intended to cover any variation, use or modification of the present invention that follows the principles of the present invention as a whole. The present invention includes the following deviations from the present disclosure that are within the known or customary practice in the art to which the present invention belongs, and that may be applied to the basic features set forth above, and within the scope of the appended claims.

Claims

1. A compound of formula (Ia), formula (Ib), formula (IIa) or formula (IIb): or a pharmaceutically acceptable salt thereof: in: m is 0, 1, 2 or 3; Selected from the group consisting of: Y is N or CH; Each R 1 are independently halogen, -CN, C1-C3 alkyl, haloalkyl, alkyl-OH, -N(R 4 )2, -NR 4 C(O)C1-C3 alkyl, -OH, -O-alkyl or cycloalkyl; R 2 is hydrogen or halogen; R 3 is hydrogen, halogen or C1-C3 alkyl; and Each R 4 are independently hydrogen or C1-C3 alkyl.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein yes 3. The compound according to claim 2 or a pharmaceutically acceptable salt thereof, wherein m is 1.

4. The compound according to claim 1, wherein R 1 It is chlorine or fluorine.

5. The compound according to claim 1, wherein The R 1 It is trifluoromethyl.

6. The compound according to claim 1, wherein The -N(R 4 )2 each R 4 The radical is hydrogen.

7. The compound according to claim 1, wherein The -N(R 4 )2 each R 4 The group is a C1-C3 alkyl group.

8. The compound according to claim 1, wherein R 1 It's hydroxymethyl.

9. The compound according to claim 1, wherein R 1 It is cyclopropyl.

10. The compound according to claim 2, wherein m is 2, and each R 1 is independently cyano, halogen, C1-C3 alkyl or cycloalkyl.

11. The compound according to claim 10, wherein Each R 1 It is a halogen.

12. The compound according to claim 10, wherein One R 1 is cyano, another R 1 is hydroxy, hydroxyalkyl, halogen, haloalkyl, alkoxy, C1-C3 alkyl, -N(R 4 )2, -NR 4 C(O)C1-C3 alkyl or cycloalkyl.

13. The compound according to claim 1, wherein yes And each R 1 It is hydrogen or methyl.

14. The compound according to claim 13, wherein Each R 1 It's methyl.

15. The compound according to any one of claims 1 to 14, wherein R 2 It is a halogen.

16. The compound according to claim 15, wherein Halogen is chlorine.

17. A compound according to any one of claims 1 to 14, wherein R 3 It is halogen or C1-C3 alkyl.

18. The compound according to claim 17, wherein The halogen is fluorine.

19. The compound according to claim 17, wherein The C1-C3 alkyl group is a methyl group.

20. The compound according to claim 1, wherein The compound is selected from the group consisting of: and pharmaceutically acceptable salts thereof.

21. A pharmaceutical composition, characterized in that Comprising an effective amount of a compound according to any one of claims 1 to 14 and a pharmaceutically acceptable excipient.

22. Use of a compound according to claim 1 alone or in combination with a pharmaceutically acceptable carrier, excipient or diluent for the preparation of a medicament for treating a patient suffering from cerebral cavernous malformation syndrome (CCM).

23. Use of the compound according to claim 1 alone or in combination with a pharmaceutically acceptable carrier, excipient or diluent in the preparation of a medicament for treating cardiovascular diseases associated with increased vascular tone.

24. The use according to claim 23, wherein The cardiovascular disease is hypertension, atherosclerosis, ischemic stroke, coronary artery spasm, cerebral vasospasm, angina pectoris or erectile dysfunction.

25. Use of the compound of claim 1 alone or in combination with a pharmaceutically acceptable carrier, excipient or diluent in the preparation of a medicament for treating a disease involving increased contractility of non-vascular smooth muscle.

26. The use according to claim 25, wherein Diseases involving increased contractility of non-vascular smooth muscle are asthma or glaucoma.

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