Deuterated Compounds as ROCK Inhibitors

Abstract

To provide a compound for treating intraocular pressure rise.SOLUTION: The solution is a compound selected from the following or a pharmaceutically acceptable salt thereof.SELECTED DRAWING: None

Description

Detailed Description of the Invention

[0001] [1] This application claims priority to U.S. Provisional Application Nos. 62 / 647,581 and 62 / 711,375, each of which is incorporated by reference in its entirety.

[0002] [Field of the Invention] [2] Certain compounds, or pharma- ceutically acceptable salts thereof, are provided that are capable of inhibiting the enzymatic activity of Rho-associated kinase (ROCK) and may be useful in the treatment of diseases and disorders mediated by ROCK.

[0003] [Background of the invention] [3] Glaucoma, as an optic neuropathy, is one of the leading causes of irreversible blindness affecting more than 60.5 million people worldwide, and is expected to affect nearly 80 million people by 2020. In glaucoma, progressive death of retinal ganglion cells leads to optic nerve damage and, ultimately, visual field loss. It is now widely accepted that elevated intraocular pressure (IOP) is the main modifiable risk factor for glaucoma, and lowering IOP is the only therapeutic intervention. In healthy eyes, aqueous humor (AH) drains primarily through the trabecular meshwork (TM), maintaining IOP in the normal range. In glaucomatous eyes, elevated IOP occurs, in part, as a result of TM dysfunction, including increased contractile tone and stiffness of the TM, altered extracellular matrix deposition, and altered permeability of the inner wall of Schlemm's canal.

[0004] [4] Novel glaucoma therapeutics are urgently needed to efficiently target the diseased trabecular pathway. Inhibitors of Rho-associated kinase (ROCK) represent a new class of targeted therapy to treat glaucoma, such as the recently FDA-approved Rhopressa (netarsudil eye drop). ROCK includes two isoforms, ROCK1 and ROCK2, which are serine / threonine kinases and downstream effectors of the small GTP-binding protein Rho. ROCK is widely expressed in various types of tissues, including TM. ROCK drives the assembly of actin stress fibers and focal adhesions, and pleiotropically regulates important cellular functions, including cell contraction, motility, morphology, polarity, cell division, and gene expression. Activation of the Rho / ROCK signaling pathway in TM and Schlemm's canal cells leads to actomyosin contraction, enhanced extracellular matrix production, and increased cell stiffness, resulting in decreased AH excretion and elevated IOP. Conversely, selective ROCK inhibitors attenuate cellular contraction and stiffness and decrease the expression levels of fibrosis-associated proteins, leading to relaxation of the TM, increased AH drainage, and ultimately reduced IOP.

[0005] [5] Therefore, compounds with inhibitory activity against ROCK would be a good therapeutic approach for glaucoma patients. Although ROCK inhibitors have been disclosed in the art, new ROCK inhibitors are needed that have at least one advantageous property selected from potency, stability, selectivity, toxicity, pharmacodynamic properties and pharmacokinetic properties. In this regard, a novel class of ROCK inhibitors is provided herein.

[0006] [Disclosure of the Invention] [6] Disclosed herein are certain novel deuterated compounds, pharma- ceutically acceptable salts thereof, and pharmaceutical compositions thereof, as well as their medicinal uses.

[0007] [7] In one aspect, disclosed herein is a compound of formula (I) or a pharma- ceutically acceptable salt thereof: [ka] (In the formula, R 1 is CH 3 , C.H. 2 D, C.H.D. 2 and CDs 3 Selected from; R 2 is CH 3 , C.H. 2 D, C.H.D. 2 and CDs 3 Selected from; Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , Y 6 , Y 7 , Y 9 , Y 10 , Y 11 , Y 12 , Y 14 , Y 15 and Y 16 are each independently selected from hydrogen and deuterium; Each Y 8 are independently selected from hydrogen and deuterium, with the proviso that both Y 8 is the same; Each Y 13 are independently selected from hydrogen and deuterium, with the proviso that both Y 13 are the same, However, the compound of formula (I) contains at least one deuterium atom.

[0008] [8] In another aspect, disclosed herein is a pharmaceutical composition comprising a compound of formula (I) or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable excipient.

[0009] [9] In yet another aspect, disclosed herein is a method of modulating ROCK, comprising administering to a system or subject in need thereof a therapeutically effective amount of a compound of formula (I) or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof, thereby modulating said ROCK.

[0010]

[10] In yet another aspect, a method of treating, ameliorating or preventing a condition responsive to inhibition of ROCK is disclosed, comprising administering to a system or subject in need of such treatment an effective amount of a compound of formula (I) or a pharma- ceutically acceptable salt thereof or a pharmaceutical composition thereof, optionally in combination with a second therapeutic agent, thereby treating said condition.

[0011]

[11] Alternatively, disclosed herein is the use of a compound of formula (I) or a pharma- ceutically acceptable salt thereof in the manufacture of a medicament for treating a condition mediated by ROCK. In certain embodiments, the compounds of the present disclosure can be used alone or in combination with a second therapeutic agent to treat a condition mediated by ROCK.

[0012]

[12] Alternatively, there is disclosed a compound of formula (I) or a pharma- ceutically acceptable salt for treating a condition mediated by ROCK.

[0013]

[13] Specifically, the conditions herein include, but are not limited to, ocular disorders such as glaucoma, ocular hypertension, or neurodegenerative eye diseases.

[0014]

[14] Further, the present disclosure discloses a method of treating elevated intraocular pressure comprising administering to a system or subject in need of such treatment an effective amount of a compound of formula (I) or a pharma- ceutically acceptable salt thereof or a pharmaceutical composition thereof, optionally in combination with a second therapeutic agent, thereby treating said condition.

[0015]

[15] Alternatively, the present disclosure provides the use of a compound of formula (I) or a pharma- ceutically acceptable salt thereof in the manufacture of a medicament for treating elevated intraocular pressure. In certain examples, the compounds of the present disclosure may be used alone or in combination with a chemotherapeutic agent to treat eye disorders.

[0016]

[16] Specifically, the eye disorders disclosed herein include, but are not limited to, glaucoma, ocular hypertension, and neurodegenerative eye diseases.

[0017]

[17] In the above methods of using the compounds of the present disclosure, a compound of formula (I) or a pharma- ceutically acceptable salt thereof may be administered to a system including a cell or tissue, or to a subject, including a mammalian subject, such as a human or animal subject.

[0018]

[18] Certain terms 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 claimed subject matter belongs. All patents, patent applications, and published materials referenced throughout this disclosure are incorporated by reference in their entirety unless otherwise noted. In the event that there is a plurality of definitions for a term in this specification, the definitions in this section shall prevail.

[0019]

[19] It is to be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not intended to be limiting of any claimed subject matter. In this application, the use of the singular includes the plural unless expressly stated otherwise. It should be noted that as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. It should also be noted that the use of "or" means "and / or" unless expressly stated otherwise. Furthermore, the use of the term "including" and other forms such as "include," "includes," and "included" are not limiting. Similarly, the use of the term "comprising" and other forms such as "comprise," "comprises," and "comprised" are not limiting.

[0020]

[20] Definitions of standard chemical terms may be found in references including Carey and Sundberg “ADVANCED ORGANIC CHEMISTRY 4th ED.” Vols. A(2000) and B(2001), Plenum Press, New York. Conventional methods of mass spectrometry, NMR, HPLC, IR and UV / Vis spectroscopy and pharmacology within the skill of the art are used unless otherwise indicated. Unless specific definitions are provided, the nomenclature used in connection with analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein, as well as the laboratory procedures and techniques thereof, are those known in the art. Standard techniques can be used for chemical synthesis, chemical analysis, pharmaceutical preparation, formulation, and delivery, and treatment of patients. Reactions and purification techniques can be performed, for example, using manufacturer's specification kits or as commonly accomplished in the art or as described herein. The techniques and procedures described above can generally be performed in a conventional manner known in the art and as described in the various general and more specific references cited and discussed throughout this specification. Throughout the specification, groups and substituents thereof can be chosen by one skilled in the art to provide stable moieties and compounds.

[0021]

[21] Where substituents are designated by conventional chemical formulas written from left to right, they equally encompass the chemically identical substituents that result from writing the structure from right to left. 2 O is for OCH 2 is equivalent to

[0022]

[22] As used herein, the term "optionally substituted" means unsubstituted or substituted. The term "substituted" means that a hydrogen atom has been removed and replaced by a substituent. It should be understood that substitution at a given atom is limited by valence. Throughout the definitions, "C i~j" denotes a range inclusive of the endpoints, where i and j are integers and indicate the number of carbon atoms. Examples include 1~4 , C 1~10 , C 3~10 etc.

[0023]

[23] The terms "deuterium" or "D" refer to an isotope of hydrogen whose nucleus contains one proton and one neutron. When a particular position is designated as having deuterium, it is understood that the abundance of deuterium at that position is greater than the natural abundance of deuterium.

[0024]

[24] The term "deuterium substituted" refers to the replacement of one or more hydrogen atoms with the corresponding number of deuterium atoms.

[0025]

[25] The term "alkyl", used alone or in combination with other terms, refers to both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. Unless otherwise specified, "alkyl" refers to C 1~10 Refers to alkyl. For example, "C 1~6 C such as "alkyl" 1~6 is defined to include groups having 1, 2, 3, 4, 5 or 6 carbons in a linear or branched arrangement. For example, "C 1~8 "Alkyl" includes, but is not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, t-butyl, i-butyl, pentyl, hexyl, heptyl and octyl.

[0026]

[26] The term "cycloalkyl", used alone or in combination with other terms, refers to a monocyclic or bridged hydrocarbon ring system. A monocyclic cycloalkyl is a carbocyclic ring system containing 3 to 10 carbon atoms, 0 heteroatoms and 0 double bonds. Examples of monocyclic ring systems include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. A monocyclic ring may contain one or two alkylene bridges, each consisting of one, two or three carbon atoms, respectively, connecting two non-adjacent carbon atoms of the ring system. Representative examples of such bridged cycloalkyl ring systems include, but are not limited to, bicyclo[3.1.1]heptane, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, bicyclo[3.2.2]nonane, bicyclo[3.3.1]nonane, bicyclo[4.2.1]nonane, tricyclo[3.3.1.03,7]nonane, and tricyclo[3.3.1.13,7]decane (adamantane). Monocyclic and bridged cycloalkyls can be attached to the parent molecular moiety through any substitutable atom contained within the ring system.

[0027]

[27] The term "halogen" (or "halo") refers to fluorine, chlorine, bromine, and iodine.

[0028]

[28] The term "alkoxy," used alone or in combination with other terms, refers to an alkyl group singly bonded to an oxygen atom. The point of attachment of the alkoxy group to the molecule is through the oxygen atom. An alkoxy group can be represented as -O-alkyl. 1~10 The term "alkoxy" refers to alkoxy groups containing from 1 to 10 carbon atoms and having straight or branched moieties. Alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, hexyloxy and the like.

[0029]

[29] The term "cycloalkoxy", used alone or in combination with other terms, refers to a cycloalkyl group that is singly bonded to an oxygen atom. The point of attachment of the cycloalkoxy group to the molecule is through the oxygen atom. A cycloalkoxy group can be represented as -O-cycloalkyl. 3~10 "Cycloalkoxy" refers to a cycloalkoxy group containing 3 to 10 carbon atoms. Cycloalkoxy groups include, but are not limited to, cyclopropoxy, cyclobutoxy, cyclohexyloxy, and the like.

[0030]

[30] The term "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids, including inorganic or organic bases and inorganic or organic acids. Salts derived from inorganic bases may be selected from, for example, aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic, manganous, potassium, sodium, and zinc salts. Further, for example, pharmaceutically acceptable salts derived from inorganic bases may be selected from ammonium, calcium, magnesium, potassium, and sodium salts. Salts in solid form may exist in one or more crystal structures and may also be in the form of hydrates. Salts derived from pharma- ceutically acceptable organic non-toxic bases may be selected from, for example, salts of primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethyl-morpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, and tripropylamine, tromethamine.

[0031]

[31] When the compounds disclosed herein are basic, salts can be prepared using at least one pharma- ceutically acceptable non-toxic acid selected from inorganic and organic acids. Such acids can be selected from, for example, acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, and p-toluenesulfonic acid. In some embodiments, such acids can be selected from, for example, citric acid, hydrobromic acid, hydrochloric acid, maleic acid, phosphoric acid, sulfuric acid, fumaric acid, and tartaric acid.

[0032]

[32] The terms "administration of" and / or "administering" a compound or a pharma- ceutically acceptable salt should be understood to mean providing a compound or a pharma- ceutically acceptable salt thereof to an individual in need of treatment.

[0033]

[33] The term "effective amount" means that amount of a compound or pharma- ceutically acceptable salt that elicits the biological or medical response in a tissue, system, animal or human that is desired by a researcher, veterinarian, medical doctor or other clinician.

[0034]

[34] The term "composition" as used herein is intended to encompass products containing specified amounts of specified ingredients, and any products resulting directly or indirectly from the combination of specified amounts of specified ingredients. Such terms, in reference to pharmaceutical compositions, are intended to encompass products containing one or more active ingredients and one or more inactive ingredients that constitute a carrier, as well as any products resulting directly or indirectly from the combination, complexation or aggregation of any two or more of the ingredients, or from any other type of reaction or interaction of one or more of the ingredients.

[0035]

[35] The term "pharmaceutical acceptable" means compatible with the other ingredients of the formulation and not unacceptably deleterious to the recipient thereof.

[0036]

[36] The term "subject" as used herein with respect to an individual suffering from a disorder, condition, etc., encompasses mammals and non-mammals. Examples of mammals include, but are not limited to, any member of the class Mammalia: humans, non-human primates such as chimpanzees, and other ape and monkey species; domestic animals such as cows, horses, sheep, goats, pigs; domestic animals such as rabbits, dogs, and cats; laboratory animals including rodents such as rats, mice, and guinea pigs, and the like. Examples of non-mammals include, but are not limited to, birds, fish, and the like. In one embodiment of the methods and compositions provided herein, the mammal is a human.

[0037]

[37] As used herein, the terms "treat," "treating," or "treatment," and other grammatical equivalents, are intended to include alleviating, reducing, or ameliorating a disease or condition, preventing additional symptoms, improving or preventing the underlying metabolic cause of a symptom, inhibiting a disease or condition, halting the onset of a disease or condition, relieving a disease or condition, causing regression of a disease or condition, alleviating a condition caused by a disease or condition, or halting the symptoms of a disease or condition, including prevention. The term further includes achieving a therapeutic benefit and / or a prophylactic benefit. A therapeutic benefit refers to the eradication or amelioration of the underlying disorder being treated. Also, a therapeutic effect is achieved by the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder, such that an improvement is observed in the patient, although the patient may still suffer from the underlying disorder. For a prophylactic benefit, the composition may be administered to a patient at risk of developing a particular disease, or to a patient reporting one or more of the physiological symptoms of the disease, even if the patient has not been diagnosed with the disease.

[0038]

[38] The term "protecting group" or "Pg" refers to a substituent that can be commonly used to block or protect certain functional groups while reacting other functional groups of a compound. For example, an "amino protecting group" is a substituent attached to an amino group that blocks or protects the amino functionality of a compound. Suitable amino protecting groups include, but are not limited to, acetyl, trifluoroacetyl, t-butoxycarbonyl (BOC), benzyloxycarbonyl (CBZ), and 9-fluorenylmethyleneoxycarbonyl (Fmoc). Similarly, a "hydroxy protecting group" refers to a substituent of a hydroxy group that blocks or protects the hydroxy functionality. Suitable protecting groups include, but are not limited to, acetyl and silyl. A "carboxy protecting group" refers to a substituent of a carboxy group that blocks or protects the carboxy functionality. Common carboxy protecting groups include -CH 2 CH 2 SO 2 Protective groups include Ph, cyanoethyl, 2-(trimethylsilyl)ethyl, 2-(trimethylsilyl)ethoxymethyl, 2-(p-toluenesulfonyl)ethyl, 2-(p-nitrophenylsulfenyl)ethyl, 2-(diphenylphosphino)-ethyl, nitroethyl, etc. For a general description of protecting groups and their uses, see TW Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, New York, 1991.

[0039]

[39] As used herein, the term "NH protecting group" includes, but is not limited to, trichloroethoxycarbonyl, tribromoethoxycarbonyl, benzyloxycarbonyl, para-nitrobenzylcarbonyl, ortho-bromobenzyloxycarbonyl, chloroacetyl, dichloroacetyl, trichloroacetyl, trifluoroacetyl, phenylacetyl, formyl, acetyl, benzoyl, tert-amyloxycarbonyl, tert-butoxycarbonyl, para-methoxybenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 4-(phenylazo)-benzyloxycarbonyl, 2-furfuryloxycarbonyl, diphenylmethoxycarbonyl, 1,1-dimethylpropoxycarbonyl, isopropoxycarbonyl, phthaloyl, succinyl, alanyl, leucyl, 1 -adamantyloxycarbonyl, 8-quinolyloxycarbonyl, benzyl, diphenylmethyl, triphenylmethyl, 2-nitrophenylthio, methanesulfonyl, paratoluenesulfonyl, N,N-dimethylaminomethylene, benzylidene, 2-hydroxybenzylidene, 2-hydroxy-5-chlorobenzylidene, 2-hydroxy-1-naphthylmethylene, 3-hydroxy-4-pyridylmethylene, cyclohexylidene, 2-ethoxycarbonylcyclohexylidene, 2-ethoxycarbonylcyclopentylidene, 2-acetylcyclohexylidene, 3,3-dimethyl-5-oxycyclo-hexylidene, diphenylphosphoryl, dibenzylphosphoryl, 5-methyl-2-oxo-2H-1,3-dioxol-4-yl-methyl, trimethylsilyl, triethylsilyl and triphenylsilyl.

[0040]

[40] As used herein, the term "C(O)OH protecting group" includes, but is not limited to, methyl, ethyl, n-propyl, isopropyl, 1,1-dimethylpropyl, n-butyl, tert-butyl, phenyl, naphthyl, benzyl, diphenylmethyl, triphenylmethyl, para-nitrobenzyl, para-methoxybenzyl, bis(para-methoxyphenyl)methyl, acetylmethyl, benzoylmethyl, para-nitrobenzoylmethyl, para-bromobenzoylmethyl, para-methanesulfonylbenzoylmethyl, 2-tetrahydropyranyl, 2-tetrahydrofuranyl, 2,2,2-trichloro-ethyl, 2-(trimethylsilyl)ethyl, acetoxymethyl, proline, ethyl ether ... These include pionyloxymethyl, pivaloyloxymethyl, phthalimidomethyl, succinimidomethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methoxymethyl, methoxyethoxymethyl, 2-(trimethylsilyl)ethoxymethyl, benzyloxymethyl, methylthiomethyl, 2-methylthioethyl, phenylthiomethyl, 1,1-dimethyl-2-propenyl, 3-methyl-3-butenyl, allyl, trimethylsilyl, triethylsilyl, triisopropylsilyl, diethylisopropylsilyl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, diphenylmethylsilyl and tert-butylmethoxyphenylsilyl.

[0041]

[41] As used herein, the term "OH or SH protecting group" includes, but is not limited to, benzyloxycarbonyl, 4-nitrobenzyloxycarbonyl, 4-bromobenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl, 1,1-dimethylpropoxycarbonyl, isopropoxycarbonyl, isobutyloxycarbonyl, diphenylmethoxycarbonyl, nyl, 2,2,2-trichloroethoxycarbonyl, 2,2,2-tribromoethoxycarbonyl, 2-(trimethylsilyl)ethoxycarbonyl, 2-(phenylsulfonyl)ethoxycarbonyl, 2-(triphenylphosphonio)ethoxycarbonyl, 2-furfuryloxycarbonyl, 1-adamantyloxycarbonyl, vinyloxycarbonyl, allyloxycarbonyl, 4-ethoxy-1-naphthyloxycarbonyl, 8-quinolyloxycarbonyl, acetyl, formyl, chloroacetyl, dichloroacetyl, These include, for example, dichloroacetyl, trichloroacetyl, trifluoroacetyl, methoxyacetyl, phenoxyacetyl, pivaloyl, benzoyl, methyl, tert-butyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, 1,1-dimethyl-2-propenyl, 3-methyl-3-butenyl, allyl, benzyl (phenylmethyl), para-methoxybenzyl, 3,4-dimethoxybenzyl, diphenylmethyl, triphenylmethyl, tetrahydrofuryl, tetrahydropyranyl, tetrahydrothiopyranyl, methoxymethyl, methylthiomethyl, benzyloxymethyl, 2-methoxyethoxymethyl, 2,2,2-trichloroethoxymethyl, 2-(trimethylsilyl)ethoxymethyl, 1-ethoxyethyl, methanesulfonyl, para-toluenesulfonyl, trimethylsilyl, triethylsilyl, triisopropylsilyl, diethylisopropylsilyl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, diphenylmethylsilyl, and tert-butylmethoxyphenylsilyl.

[0042]

[42] The compounds may exist as geometric isomers. The compounds of the invention may contain carbon-carbon or carbon-nitrogen double bonds in the E or Z configuration, with the term "E" representing the higher order substituents on opposite sides of the carbon-carbon or carbon-nitrogen double bond and the term "Z" representing the higher order substituents on the same side of the carbon-carbon or carbon-nitrogen double bond, as determined by the Cahn-Ingold-Prelog precedence rules. The compounds of the invention may also exist as mixtures of "E" and "Z" isomers. Substituents around a cycloalkyl or heterocycloalkyl are designated to be in the cis or trans configuration. Additionally, the invention contemplates various isomers and mixtures thereof resulting from the arrangement of substituents around the adamantane ring system. Two substituents around a single ring within the adamantane ring system are designated as Z or E relative configuration. See, for example, CD Jones, M. Kaselj, RN Salvatore, WJ le Noble J. Org. Chem.1998, 63, 2758-2760.

[0043]

[43] The compounds of the invention can contain asymmetrically substituted carbon atoms in the R or S configuration, the terms "R" and "S" being as defined by IUPAC 1974 Recommendations for Section E, Fundamental Stereochemistry, Pure Appl. Chem. (1976) 45, 13-10. Compounds having asymmetrically substituted carbon atoms with equal amounts of R and S configurations are racemic at those carbon atoms. Atoms in excess of one configuration over the other are assigned the configuration that is present in the greater amount, preferably about 85-90% excess, more preferably about 95-99% excess, even more preferably greater than about 99% excess. Thus, the invention includes racemic mixtures, relative and absolute stereoisomers, and mixtures of relative and absolute stereoisomers.

[0044]

[44] Isotopically enriched or labeled compounds The compounds of the present invention can be present in isotopically labeled or enriched form, containing one or more atoms with atomic mass or mass number different from the atomic mass or mass number most abundantly found in nature. The isotope can be a radioactive or non-radioactive isotope. The isotopes of atoms such as hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine and iodine include, but are not limited to, 2 H, 3 H, 13 C. 14 C. 15 N, 18 O. 32 P, 35 S, 18 F, 36 Cl and 125 I. Compounds that contain other isotopes of these and / or other atoms are within the scope of this invention.

[0045]

[45] In another embodiment, the isotopically labeled compound is isotopically labeled with deuterium ( 2 H), tritium ( 3 H) or 14 C isotope. Thus, when a substituent is described as hydrogen, the substituent also incorporates isotopic equivalents such as deuterium and tritium, particularly deuterium (D). Isotopically labeled compounds of the present invention can be prepared by common methods well known to those skilled in the art. Such isotopically labeled compounds can be conveniently prepared by carrying out the procedures disclosed in the examples and schemes disclosed herein by substituting readily available isotopically labeled reagents for non-labeled reagents. In some cases, the compound can be treated with an isotopically labeled reagent to exchange a normal atom for its isotope, e.g., hydrogen for D 2 SO 4 / D 2Deuterium can be exchanged by the action of a deuterated acid such as O. In addition to the above, relevant procedures and intermediates are described, for example, in Lizondo, J et al, Drugs Fut, 21(11), 1116(1996); Brickner, SJ et al., J Med Chem, 39(3), 673(1996); Mallesham, B et al, Org Lett, 5(7), 963 (2003); PCT Publication, WO 1997010223, WO 2005099353, WO 1995007271, WO 2006008754; U.S. Patent Nos. 7538189; 7534814; 7531685; 7528131; 7521421; 7514068; 7511013; and U.S. Patent Application Publication No. 20090137457. Nos. 20090131485; 20090131363; 20090118238; 20090111840; 20090105338; 20090105307; 20090105147; 20090093422; 20090088416; and 20090082471, the methods of which are incorporated herein by reference.

[0046]

[46] The isotope-labeled compounds of the present invention can be used as standards to determine the efficacy of STAT3 inhibitors in binding assays.Isotopically-containing compounds have been used in pharmaceutical research to investigate the in vivo metabolic fate of compounds by evaluating the mechanism of action and metabolic pathways of non-isotopically-labeled parent compounds (Blake et al. J.Pharm.Sci.64,3,367-391(1975)). Such metabolic studies are important in the design of safe and effective therapeutic agents, as metabolites produced from in vivo active compounds or parent compounds administered to patients may prove to be toxic or carcinogenic (Foster et al., Advances in Drug Research Vol. 14, pp. 2-36, Academicpress, London, 1985; Kato et al, J. Labelled Comp. Radiopharmaceut., 36(10): 927-932(1995);Kushner et al., Can. J. Physiol. Pharmacol, 77, 79-88(1999)).

[0047]

[47] Additionally, non-radioactive isotope-containing drugs, such as deuterated drugs, referred to as "heavy drugs," can be used to treat diseases and conditions associated with STAT3 activity. Increasing the amount of an isotope present in a compound above its natural abundance is called enrichment. Examples of amounts of enrichment include about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 16, 21, 25, 29, 33, 37, 42, 46, 50, 54, 58, 63, 67, 71, 75, 79, 84, 88, 92, 96 to about 100 mol%. In mammals, including rodents and dogs, replacement of up to about 15% of normal atoms with heavier isotopes has been observed for periods of days to weeks with minimal adverse effects (Czajka DM and Finkel AJ, Ann. NY Acad. Sci. 1960 84: 770; Thomson J F, Ann. New York Acad. Sci 1960 84: 736; Czakja DM et al., Am. J. Physiol. 1961201: 357). Acute replacement of human body fluids with deuterium at 15%-23% has been shown to cause no toxicity (Blagojevic N et al. in “Dosimetry&Treatment Planning for Neutron Capture Therapy”, Zamenhof R, Solares G and Harling O Eds. 1994. Advanced Medical Publishing, Madison Wis. pp.125-134; Diabetes Metab. 23: 251(1997)).

[0048]

[48] ​​Stable isotope labeling of drugs can alter physicochemical properties such as pKa and lipid solubility. If the isotope substitution affects regions involved in ligand-receptor interactions, these effects and changes can affect the pharmacodynamic response of the drug molecule. Some of the physical properties of stable isotope-labeled molecules differ from non-labeled molecules, while the chemical and biological properties are the same, with one important exception: due to the increased mass of the heavy isotope, a bond involving the heavy isotope and another atom will be stronger than the same bond between that atom and a light isotope. Thus, incorporation of an isotope at a site of metabolism or enzymatic conversion can slow down said reactions and alter the pharmacokinetic profile or efficacy compared to the non-isotopic compound.

[0049]

[49] In one embodiment, the present invention provides a method for producing [ka] [ka] and a pharma- ceutically acceptable salt thereof.

[0050]

[50] In embodiment (1), the present invention provides a compound of formula (I) or a pharma- ceutically acceptable salt thereof: [ka] (In the formula, R 1 is CH 3 , C.H. 2 D, C.H.D. 2 and CDs 3 is selected from R 2 is CH 3 , C.H. 2 D, C.H.D. 2 and CDs 3 is selected from Y 1 , Y 2 , Y 3 , Y4 , Y 5 , Y 6 , Y 7 , Y 9 , Y 10 , Y 11 , Y 12 , Y 14 , Y 15 and Y 16 are each independently selected from hydrogen and deuterium; Each Y 8 are independently selected from hydrogen and deuterium, with the proviso that both Y 8 are the same, Each Y 13 are independently selected from hydrogen and deuterium, with the proviso that both Y 13 are the same, However, the compound of formula (I) contains at least one deuterium atom.

[0051]

[51] In another embodiment (2), the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis, comprising the steps of: 1 is deuterium.

[0052]

[52] In another embodiment (3), the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis, comprising the steps of: 2 The compound according to any one of embodiments (1)-(2), or a pharma- ceutically acceptable salt thereof, is provided, wherein is deuterium.

[0053]

[53] In another embodiment (4), the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis, comprising the steps of: 3 The compound of any one of embodiments (1)-(3), or a pharma- ceutically acceptable salt thereof, is provided, wherein is deuterium.

[0054]

[54] In another embodiment (5), the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis, comprising the steps of: 7 The compound according to any one of embodiments (1)-(4), or a pharma- ceutically acceptable salt thereof, is provided, wherein is deuterium.

[0055]

[55] In another embodiment (6), the present invention provides a method for producing a medicament comprising the steps of: 8 The compound according to any one of embodiments (1)-(5), or a pharma- ceutically acceptable salt thereof, is provided, wherein is deuterium.

[0056]

[56] In another embodiment (7), the present invention provides both Y 13 The compound of any one of embodiments (1)-(6), or a pharma- ceutically acceptable salt thereof, is provided, wherein is deuterium.

[0057]

[57] In another embodiment (8), the present invention provides a compound comprising R 1 CH 3 and CDs 3 or a pharma- ceutically acceptable salt thereof.

[0058]

[58] In another embodiment (9), the present invention provides a compound comprising R 2 CH 3 and CDs 3 or a pharma- ceutically acceptable salt thereof.

[0059]

[59] In another embodiment (10), the present invention provides a method for producing a composition comprising: [ka] and a pharma- ceutically acceptable salt thereof.

[0060]

[60] In another embodiment (11), the present invention provides the compound of embodiment (10) or a pharma- ceutically acceptable salt thereof, [ka] and pharma- ceutically acceptable salts thereof.

[0061]

[61] In another embodiment (12), the present invention provides the compound of embodiment (11) or a pharma- ceutically acceptable salt thereof, [ka] and pharma- ceutically acceptable salts thereof.

[0062]

[62] In another embodiment (13), the present invention provides a compound of any one of embodiments (1) to (12) or a pharma- ceutically acceptable salt thereof, which is the hydrochloride salt.

[0063]

[63] In another embodiment (14), the present invention provides a compound of embodiment (13) or a pharma- ceutically acceptable salt thereof, which is the dihydrochloride salt.

[0064]

[64] In another embodiment (15), the present invention relates to a compound of embodiment (14) or [ka] JPEG0007652865000009.jpg73149 and a pharma- ceutically acceptable salt thereof selected from:

[0065]

[65] In another embodiment (16), the present invention provides a pharmaceutical composition comprising a compound of any one of embodiments (1) to (15) or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable carrier.

[0066]

[66] In another embodiment (17), the present invention provides a method for treating, ameliorating, or preventing a condition responsive to inhibition of ROCK, comprising the step of administering to a subject in need of such treatment an effective amount of a compound of any one of embodiments (1)-(15) or a pharma- ceutically acceptable salt thereof or a pharmaceutical composition thereof, optionally in combination with a second therapeutic agent.

[0067]

[67] In another embodiment (18), the present invention provides the use of a compound of any one of embodiments (1) to (16), or a pharma- ceutically acceptable salt thereof, in the preparation of a medicament for treating elevated intraocular pressure.

[0068]

[68] In yet another of its aspects, a kit is provided that includes a compound disclosed herein, or a pharma- ceutically acceptable salt thereof; and instructions that include one or more forms of information selected from the group consisting of instructions for the condition for which the composition is to be administered, storage information for the composition, dosing information, and instructions regarding how to administer the composition. In one particular form, the kit includes a multiple dose form of the compound.

[0069]

[69] In yet another of its aspects, an article of manufacture is provided that includes a compound disclosed herein or a pharma- ceutically acceptable salt thereof; and packaging material. In one variation, the packaging material includes a container for housing the compound. In one particular variation, the container includes a label that indicates one or more members of the group consisting of a medical condition for which the compound is to be administered, storage information, dosing information, and / or instructions regarding how to administer the compound. In another variation, the article of manufacture includes the compound in a multiple dose form.

[0070]

[70] In a further aspect thereof, there is provided a method of treatment comprising administering a compound disclosed herein or a pharma- ceutically acceptable salt thereof.

[0071]

[71] In another of its aspects, there is provided a method of inhibiting ROCK, comprising contacting ROCK with a compound disclosed herein, or a pharma- ceutically acceptable salt thereof.

[0072]

[72] In yet another of its aspects, there is provided a method of inhibiting ROCK, comprising the step of causing a compound disclosed herein, or a pharma- ceutically acceptable salt thereof, to be present in a subject, so as to inhibit ROCK in vivo.

[0073]

[73] In a further aspect thereof, there is provided a method of inhibiting ROCK comprising administering to a subject a first compound that is converted in vivo to a second compound, wherein the second compound inhibits ROCK in vivo, and the second compound is a compound according to any one of the above embodiments and variations.

[0074]

[74] In another of its aspects, there is provided a method of treating a condition in which ROCK has activity that contributes to the pathology and / or symptomatology of the condition, comprising the step of presenting to a subject a compound disclosed herein, or a pharma- ceutically acceptable salt thereof, in an amount therapeutically effective for the condition.

[0075]

[75] In a further aspect thereof, there is provided a method of treating a condition in which ROCK has activity that contributes to the pathology and / or symptomatology of the condition, comprising administering to a subject a first compound that is converted in vivo to a second compound, where the second compound inhibits ROCK in vivo. Note that a compound of the invention can be the first or second compound.

[0076]

[76] In one variation of each of the above methods, the condition is selected from an ocular disorder such as glaucoma, ocular hypertension, or a neurodegenerative eye disease.

[0077]

[77] In another of its aspects, a method is provided for treating a condition in which a mutation in a ROCK gene contributes to the pathology and / or symptomatology of the condition, including, for example, glaucoma, ocular hypertension, or neurodegenerative eye diseases.

[0078]

[78] In yet another of its aspects, the invention relates to the use of a compound of any of the above embodiments and variations as a medicament. In yet another of its aspects, the invention relates to the use of a compound according to any one of the above embodiments and variations in the manufacture of a medicament for inhibiting ROCK.

[0079]

[79] In a further of its aspects, the invention relates to the use of a compound according to any one of the above embodiments and variations in the manufacture of a medicament for treating a condition in which ROCK has activity that contributes to the pathology and / or symptomatology of the condition.

[0080]

[80] Administration and Pharmaceutical Compositions Generally, the compound of the present disclosure is administered in a therapeutically effective amount by itself or in combination with one or more therapeutic agents through any of the usual acceptable methods known in the art.The therapeutically effective amount can vary widely depending on the severity of disease, the age and relative health of the subject, the potency of the compound used, and other factors known to those skilled in the art.For example, for the treatment of neoplastic disease and immune system disorder, the required dosage will also vary depending on the mode of administration, the specific condition to be treated and the desired effect.

[0081]

[81] In general, satisfactory results have been shown to be obtained systemically at daily doses of about 0.001 to about 100 mg / kg body weight, or in particular about 0.03 to 2.5 mg / kg body weight. The daily dose indicated for larger mammals, e.g., humans, is suitably administered in the range of about 0.5 mg to about 2000 mg, more particularly about 0.5 mg to about 1000 mg, and may be administered, for example, in divided doses up to four times a day or in delayed form. Unit dosage forms suitable for oral administration contain about 1 to 50 mg of active ingredient.

[0082]

[82] The compounds of the present disclosure may be administered as a pharmaceutical composition by any conventional route; for example, enterally, for example, orally, for example, in the form of tablets or capsules; parenterally, for example, in the form of injectable solutions or suspensions; or topically, for example, in the form of eye drops, lotions, gels, ointments or creams, or in the nasal or suppository form.

[0083]

[83] Pharmaceutical compositions comprising the compounds of the present disclosure in free form or in pharma- ceutically acceptable salt form together with at least one pharma- ceutically acceptable carrier or diluent can be prepared in a conventional manner by mixing, granulating, coating, dissolving or lyophilizing processes. For example, pharmaceutical compositions comprising the compounds of the present disclosure together with at least one pharma- ceutically acceptable carrier or diluent can be prepared in a conventional manner by mixing with a pharma- ceutically acceptable carrier or diluent. A unit dosage form for oral administration contains, for example, about 0.1 mg to about 500 mg of active substance.

[0084]

[84] In one embodiment, the pharmaceutical composition is a solution of the active ingredient, including a suspension or dispersion, such as an isotonic aqueous solution. In the case of a lyophilized composition, including the active ingredient alone or together with a carrier, such as mannitol, a dispersion or suspension can be prepared before use. The pharmaceutical composition may be sterilized and / or may contain adjuvants, such as preservatives, stabilizers, wetting agents or emulsifiers, dissolution enhancers, salts for adjusting osmotic pressure and / or buffers. Suitable preservatives include, but are not limited to, antioxidants, such as ascorbic acid, or microbicides, such as sorbic acid or benzoic acid. The solution or suspension may further include a thickener, including, but not limited to, sodium carboxymethylcellulose, carboxymethylcellulose, dextran, polyvinylpyrrolidone, gelatin, or a solubilizer, for example, Tween 80 (polyoxyethylene (20) sorbitan monooleate).

[0085]

[85] Suspensions in oil may contain as the oil component a vegetable oil, synthetic oil, or semi-synthetic oil conventional for injection purposes. Examples include liquid fatty acid esters containing as the acid component a long-chain fatty acid having 8 to 22 carbon atoms, or in some embodiments 12 to 22 carbon atoms. Suitable liquid fatty acid esters include, but are not limited to, lauric acid, tridecylic acid, myristic acid, pentadecylic acid, palmitic acid, margaric acid, stearic acid, arachidic acid, behenic acid or the corresponding unsaturated acids, such as oleic acid, elaidic acid, erucic acid, brassidic acid and linoleic acid, and may contain, if desired, an antioxidant, such as vitamin E, 3-carotene or 3,5-di-tert-butyl-hydroxytoluene. The alcohol component of these fatty acid esters may have 6 carbon atoms and may be a monohydric or polyhydric, e.g., monohydric, dihydric or trihydric alcohol. Suitable alcohol components include, but are not limited to, methanol, ethanol, propanol, butanol or pentanol or their isomers; glycol and glycerol.

[0086]

[86] Other suitable fatty acid esters include, but are not limited to, ethyl oleate, isopropyl myristate, isopropyl palmitate, LABRAFIL® M 2375, (polyoxyethylene glycerol), LABRAFIL® M 1944 CS (unsaturated polyglycolized glycerides including glycerides and polyethylene glycol esters prepared by alcoholysis of apricot kernel oil), LABRASOL™ (saturated polyglycolized glycerides including glycerides and polyethylene glycol esters prepared by alcoholysis of TCM, all available from GaKefosse, France), and / or MIGLYOL® 812 (triglycerides of saturated fatty acids with chain lengths of C8 to C12 manufactured by Huls AG, Germany), as well as vegetable oils such as cottonseed oil, almond oil, olive oil, castor oil, sesame oil, soybean oil, or peanut oil.

[0087]

[87] Pharmaceutical compositions for oral administration can be obtained, for example, by combining the active ingredient with one or more solid carriers, optionally granulating the resulting mixture, and processing the mixture or granules by including additional excipients to form tablets or tablet cores.

[0088]

[88] Suitable carriers include, but are not limited to, sugars such as lactose, saccharose, mannitol or sorbitol, cellulose preparations, and / or fillers such as calcium phosphates, e.g. tricalcium phosphate or calcium hydrogen phosphate, and also binders such as starches, e.g. corn, wheat, rice or potato starch, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidone, and / or disintegrating agents, if desired, such as the above-mentioned starches, carboxymethyl starch, cross-linked polyvinylpyrrolidone, alginic acid or a salt thereof, e.g. sodium alginate. Additional excipients include flow conditioners and lubricants, e.g. silicic acid, talc, stearic acid or a salt thereof, e.g. magnesium stearate or calcium stearate, and / or polyethylene glycol, or derivatives thereof.

[0089]

[89] The tablet cores may be provided with a suitable, optionally enteric, coating through the use of concentrated sugar solutions, which may contain, inter alia, gum arable, talc, polyvinylpyrrolidone, polyethylene glycol and / or titanium dioxide, or coating solutions in suitable organic solvents or solvent mixtures, or to prepare an enteric coating, a solution of a suitable cellulose preparation, for example acetylcellulose phthalate or hydroxypropylmethylcellulose phthalate. Dyes or pigments may be added to the tablets or tablet coatings, for example for identification purposes or to indicate different doses of active ingredient.

[0090]

[90] Pharmaceutical compositions for oral administration may also include hard capsules containing gelatin or soft sealed capsules containing gelatin and a plasticizer such as glycerol or sorbitol. Hard capsules may contain the active ingredient in the form of granules, for example, mixed with fillers such as corn starch, binders, and / or lubricants such as talc or magnesium stearate, and optionally stabilizers. In soft capsules, the active ingredient is dissolved or suspended in a suitable liquid excipient, such as fatty oils, paraffin oil or liquid polyethylene glycols or fatty acid esters of ethylene or propylene glycol, to which stabilizers and surfactants, for example polyoxyethylene sorbitan fatty acid ester types, may be added.

[0091]

[91] Pharmaceutical compositions suitable for rectal administration are, for example, suppositories that contain a combination of the active ingredient and a suppository base. Suitable suppository bases are, for example, natural or synthetic triglycerides, paraffin hydrocarbons, polyethylene glycols or higher alkanols.

[0092]

[92] Pharmaceutical compositions suitable for parenteral administration may comprise aqueous solutions of the active ingredient in water-soluble form, e.g., as a water-soluble salt, or aqueous injection suspensions containing viscosity-increasing substances, e.g., sodium carboxymethylcellulose, sorbitol and / or dextran, and, if desired, stabilizers. The active ingredient, optionally together with excipients, may be in the form of a lyophilizate and made into a solution before parenteral administration by the addition of suitable solvents. Solutions such as those used for parenteral administration, for example, may also be used as infusion solutions. The manufacture of preparations for injection is usually carried out under aseptic conditions, as is, for example, the filling into ampoules or vials and the sealing of the containers.

[0093]

[93] The present disclosure also provides pharmaceutical combinations, e.g., kits, comprising: a) a first agent that is a compound of the present disclosure disclosed herein, in free form or in a pharma- ceutically acceptable salt form; and b) at least one co-agent. The kit can include instructions for administration.

[0094]

[94] Combination therapy The compounds or pharma- ceutically acceptable salts of the present disclosure may be administered as monotherapy or in combination with one or more other therapeutic agents.

[0095]

[95] For example, the therapeutic effectiveness of one of the compounds described herein may be enhanced by administration of an adjuvant (i.e., the adjuvant itself may have minimal therapeutic benefit, but in combination with another therapeutic agent, the overall therapeutic benefit to the individual is enhanced). Alternatively, by way of example only, the benefit experienced by an individual may be increased by administering one of the compounds described herein with another therapeutic agent that also has a therapeutic benefit. By way of example only, in the treatment of gout involving administration of one of the compounds described herein, an increased therapeutic benefit may be provided by providing the individual with another therapeutic agent for gout as well. Alternatively, by way of example only, if one of the side effects experienced by an individual upon receiving one of the compounds described herein is nausea, it may be appropriate to administer an anti-nausea agent in combination with the compound. Alternatively, one or more additional therapies may include, but are not limited to, physical therapy, psychotherapy, radiation therapy, application of compresses to the affected area, rest, dietary changes, and the like. Regardless of the disease, disorder, or condition being treated, the overall benefit experienced by the individual may be additive of the two therapies, or the individual may experience a synergistic benefit.

[0096]

[96] When the compounds described herein are administered in combination with other therapeutic agents, the compounds described herein may be administered in the same pharmaceutical composition as the other therapeutic agents, or may be administered by different routes due to different physical and chemical properties. For example, the compounds described herein may be administered orally to generate and maintain good blood levels, while the other therapeutic agents are administered intravenously. Thus, the compounds described herein may be administered simultaneously, sequentially, or separately with the other therapeutic agents. EXAMPLES

[0097]

[97] Various methods can be developed to synthesize the compound of formula (I) or its pharma- ceutically acceptable salt. Representative methods for synthesizing the compound of formula (I) or its pharma- ceutically acceptable salt are provided in the examples. However, it should be noted that the compound of formula (I) or its pharma- ceutically acceptable salt can also be synthesized by other synthetic routes that others may devise.

[0098]

[98] It will be readily recognized that certain compounds of formula (I) have atoms with bonds to other atoms that confer a particular stereochemistry to the compound (e.g., chiral centers). It will be recognized that synthesis of compounds of formula (I) or pharma- ceutically acceptable salts thereof may result in the production of mixtures of different stereoisomers (enantiomers, diastereomers). Unless a particular stereochemistry is specified, recitation of the compound is intended to encompass all of the various possible stereoisomers.

[0099]

[99] The compounds of formula (I) can also be prepared as pharma- ceutically acceptable acid addition salts, for example, by reacting the free base form of at least one compound with a pharma- ceutically acceptable inorganic or organic acid. Alternatively, pharma- ceutically acceptable base addition salts of at least one compound of formula (I) can be prepared, for example, by reacting the free acid form of at least one compound with a pharma- ceutically acceptable inorganic or organic base. Inorganic and organic acids and bases suitable for preparing pharma- ceutical acceptable salts of compounds of formula (I) are set forth in the definitions section of this application. Alternatively, salt forms of compounds of formula (I) can be prepared using salts of starting materials or intermediates.

[0100]

[0100] The free acid or free base form of the compound of formula (I) can be prepared from the corresponding base addition salt or acid addition salt form. For example, the compound of formula (I) in the form of an acid addition salt can be converted to its corresponding free base by treating with a suitable base (e.g., ammonium hydroxide solution, sodium hydroxide, etc.). The compound of formula (I) in the form of a base addition salt can be converted to its corresponding free acid by treating with a suitable acid (e.g., hydrochloric acid, etc.).

[0101]

[0101] The N-oxide of the compound of formula (I) or a pharma- ceutically acceptable salt thereof can be prepared by methods known to those skilled in the art. For example, the N-oxide can be prepared by treating the non-oxidized form of the compound of formula (I) with an oxidizing agent (e.g., trifluoroperacetic acid, permaleic acid, perbenzoic acid, peracetic acid, meta-chloroperbenzoic acid, etc.) in a suitable inert organic solvent (e.g., a halogenated hydrocarbon such as dichloromethane) at about 0-80°C. Alternatively, the N-oxide of the compound of formula (I) can be prepared from the N-oxide of a suitable starting material.

[0102]

[0102] The non-oxidized form of the compound of formula (I) can be prepared from the N-oxide of the compound of formula (I), for example, by treatment with a reducing agent (e.g., sulfur, sulfur dioxide, triphenylphosphine, lithium borohydride, sodium borohydride, phosphorus trichloride, phosphorus tribromide, etc.) in a suitable inert organic solvent (e.g., acetonitrile, ethanol, aqueous dioxane, etc.) at 0-80°C.

[0103]

[0103] Protected derivatives of compounds of formula (I) can be prepared by methods known to those skilled in the art. A detailed description of the techniques applicable to the creation of protecting groups and their removal can be found in TW Greene, Protecting Groups in Organic Synthesis, 3rd edition, John Wiley & Sons, Inc. 1999.

[0104]

[0104] As used herein, the symbols and notations used in these processes, schemes, and examples are consistent with those used in the contemporary scientific literature, e.g., the Journal of the American Chemical Society or the Journal of Biological Chemistry. Standard one-letter or three-letter abbreviations are generally used to designate amino acid residues, which are assumed to be in the L-configuration unless otherwise noted. Unless otherwise noted, all starting materials were obtained from commercial suppliers and used without further purification. For example, the following abbreviations may be used in the examples and throughout the specification: g (grams); mg (milligrams); L (liters); mL (milliliters); μL (microliters); psi (pounds per square inch); M (molar); mM (millimolar); iv (intravenous); Hz (hertz); MHz (megahertz); mol (mole); mmol (millimolar); RT (room temperature); min (minutes); h (hours); mp (melting point); TLC (thin layer chromatography); Rt (retention time); RP (reverse phase); MeOH (methanol); i-PrOH (isopropanol); TEA (triethylamine); ;TFA(trifluoroacetic acid);TFAA(trifluoroacetic anhydride);THF(tetrahydrofuran);DMSO(dimethylsulfoxide);EtOAc(ethyl acetate);DME(1,2-dimethoxyethane);DCM(dichloromethane);DCE(dichloroethane);DMF(N,N-dimethylformamide);DMPU(N,N-dimethylpropyleneurea);CDI(1,1-carbonyldiimidazole);IBCF(isobutyl chloroformate);HOAc(acetic acid);HOSu(N-hydroxysuccinimide);HOBT(1-hydroxybenzotriazole);Et 2O (diethyl ether); EDCI (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride); BOC (tert-butyloxycarbonyl); FMOC (9-fluorenylmethoxycarbonyl); DCC (dicyclohexylcarbodiimide); CBZ (benzyloxycarbonyl); Ac (acetyl); atm (atmosphere); TMSE (2-(trimethylsilyl)ethyl); TMS (trimethylsilyl); TIPS (triisopropylsilyl); TBS (t-butyldimethylsilyl); DMAP (4-dimethylaminopyridine); Me (methyl); OMe (methoxy); Et (ethyl); tBu (tert-butyl); HPLC (high pressure liquid chromatography); BOP (bis(2-oxo-3-oxazolidinyl)phosphinic chloride); TBAF (tetra-n-butylammonium fluoride); m-CPBA (metachloroperbenzoic acid).

[0105] Ether or Et 2 References to O are references to diethyl ether; brine refers to a saturated aqueous solution of NaCl. All temperatures are in °C (degrees Celsius) unless otherwise indicated. All reactions were conducted under an inert atmosphere at RT unless otherwise noted.

[0106]

[0106] 1 H NMR spectra were recorded on a Varian Mercury Plus 400. Chemical shifts are expressed in parts per million (ppm). Coupling constants are in Hertz (Hz). Splitting patterns describe the apparent multiplicity and are designated as s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), and br (broad).

[0107] Low-resolution mass spectra (MS) and compound purity data were acquired on a Shimadzu LC / MS single quadrupole system equipped with an electrospray ionization (ESI) source, UV detector (220 and 254 nm), and evaporative light scattering detector (ELSD). Thin-layer chromatography was performed on 0.25 mm Superchemgroup silica gel plates (60F-254) and visualized with UV light, 5% ethanolic phosphomolybdic acid, ninhydrin, or p-anisaldehyde solution. Flash column chromatography was performed on silica gel (200-300 mesh, branch of Qingdao Haiyang Chemical Co., Ltd).

[0108] Synthesis scheme The compound of formula (I) or a pharma- ceutically acceptable salt thereof can be synthesized according to various reaction schemes. Some exemplary schemes are provided below and in the Examples. Other reaction schemes can be easily devised by those skilled in the art in view of this disclosure.

[0109] In the reactions described below, reactive functional groups, such as hydroxyl, amino, imino, thio or carboxyl groups, may need to be protected to avoid undesired participation in the reaction, if these are desired in the final product. Conventional protecting groups can be used according to standard practice, see, for example, TW Greene and PGM Wuts in "Protective Groups in Organic Chemistry" John Wiley and Sons, 1991.

[0110]

[0110] Synthetic methods for preparing the compounds of the invention are illustrated in the following schemes and examples. Starting materials are commercially available or can be prepared according to procedures known in the art or as illustrated herein.

[0111]

[0111] The intermediates depicted in the following schemes can be prepared by a variety of methods known in the literature or familiar to those skilled in the art.

[0112]

[0112] As an example, one synthesis of the compound of formula I of the present disclosure is outlined in Scheme 1. As shown in the scheme, the compound of formula I can be resolved into intermediates II and III, which can be prepared by various methods known in the literature or familiar to those skilled in the art. In this route, intermediate II can be obtained via a three-step sequence including protection of the free carboxyl group of IV, coupling with V, and cleavage of the protecting group. Finally, coupling of the carboxylic acid II with an aminoarene, such as the aminoarene of formula III, via a condensation reaction, gives the compound of formula I. [ka]

[0113] As an illustration of the preparation of intermediates of formula IV, one synthetic route to intermediate IVa is shown in Scheme 2. Starting from commercially available IVa-A, IVa-C can be prepared by carbonyl insertion reaction and TBS protection of the free hydroxyl group of IVa-B. LiAlD 4 Reduction of ester IVa-C with IVa-D gives IVa-D, which can be further converted to IVa-E through condensation with benzoic acid VI. Finally, intermediates of formula IVa can be obtained from IVa-E through cleavage of the protecting groups on the successive oxygens and oxidation of the resulting alcohol. [ka]

[0114] As a further illustration of the preparation of intermediates of formula IV, one synthetic route to intermediate IVb is provided in Scheme 3. Commercially available dimethyl terephthalate IVb-A is selected as the starting material. LiAlD 4Reduction of IVb-A with IVb-B gives IVb-B, which can be further converted to IVb-D through selective TBDPS protection of the hydroxyl group and iodination. Nucleophilic displacement of the iodide of IVb-D with TMSCN gives intermediate IVb-E. Conversion of the cyano group of IVb-E to a methyl ester group gives intermediate of formula IVb-G. Finally, compounds of formula IVb can be prepared by condensation with benzoic acid VI followed by hydrolysis of IVb-H. [ka]

[0115] Another illustration of the preparation of intermediates of formula IV is shown in Scheme 4, which illustrates the preparation of compounds of formula IVc. Starting from commercially available 2,4-dimethylbenzoic acid IVc-A, IVc-A can be prepared by the addition of t-BuOK and DMSO-d 6 The intermediate 2,4-bis(methyl-d3)benzoic acid IVc-B can be prepared by treating with 2,4-bis(methyl-d 3 ) Coupling of benzoic acid IVc-B with intermediate of formula IVc-C via condensation reaction gives compound of formula IVc-D. Finally, hydrolysis of IVc-D gives compound of formula IVc. [ka]

[0116] As an illustration of the preparation of intermediates of formula III, one synthetic route to compound IIIa is shown in Scheme 5. Starting from commercially available 1,4-dibromobenzene IIIa-A, IIIa-A can be synthesized by the addition of n-BuLi and DMF-d 7 Aldehyde IIIa-B can be prepared by treatment with 2,2-dimethoxyethan-1-amine and NaBD 4 Reductive amination using AlCl followed by protection of the amino group of IIIa-D can give intermediate IIIa-E. 3Cyclization of IIIa-E promoted by a Lewis acid such as IIIa-F affords isoquinolines of formula IIIa-F, which can be further converted to IIIa-G via palladium-catalyzed coupling with benzophenone imine. Finally, deprotection of IIIa-G affords compounds of formula IIIa. [ka]

[0117]

[0117] As a further illustration of the preparation of intermediates of formula III, one synthetic route to compound IIIb is shown in Scheme 6. Starting from commercially available 2-oxoacetic acid IIIb-A, intermediate IIIb-B can be prepared by treating IIIb-A with trimethoxymethane and p-TsOH. Ammonolysis of IIIb-B gives compound of formula IIIb-C, which can be further purified by cycloaddition of LiAlD 4 The intermediate IIIb-D can be further converted to a compound of formula IIIb-D via reduction of the amide with 4-bromobenzaldehyde and NaBH 4 Through reductive amination using AlCl followed by protection of the amino group of IIIb-F, intermediate IIIb-G can be obtained. 3 Cyclization of IIIb-G promoted by a Lewis acid such as IIIb-H gives the isoquinoline of formula IIIb-H, which can be further converted to IIIb-I via palladium-catalyzed coupling with benzophenone imine. Finally, deprotection of IIIb-I gives the compound of formula IIIb. [ka]

[0118] As an illustration of the preparation of intermediates of formula V, one synthetic route to compound Va is shown in Scheme 7. Starting from commercially available 1H-benzo[d][1,2,3]triazole Va-A, Va-A can be reacted with formaldehyde-d 2The intermediate Va-B can be prepared by treating with . The hydroxyl group of Va-B can then be converted to a Boc-protected amino group with tert-butyl carbamate and 4-methylbenzenesulfonic acid to give the compound of formula Va. [ka]

[0119]

[0119] In some cases, the order of carrying out the above reaction schemes can be varied to facilitate the reaction or to avoid unwanted reaction products. The following examples are provided so that the present invention might be more fully understood. These examples are merely illustrative and should not be construed as limiting the invention in any way.

[0120] Intermediate A Isoquinolin-1-d-6-amine (A) [ka]

[0121] (4-Bromophenyl)-deuterio-methanone (A1)

[0122]

[0122] To a solution of 1,4-dibromobenzene (2.50 g, 10.6 mmol) in THF (5 mL) was added n-BuLi (1.6 M, 7 mL, 11.7 mmol) dropwise at -78°C, and the resulting mixture was stirred at -78°C for 0.5 h. Then, DMF-d 7 (1.08 mL, 12.7 mmol) was added dropwise to the mixture at -78 °C and the resulting mixture was stirred at -78 °C to -30 °C for 1.5 h. The reaction was diluted with saturated NH 4 The mixture was quenched with aqueous Cl (2 mL) at -30°C. The mixture was diluted with water and extracted with EtOAc (3x50 mL). The extracts were washed with brine and Na 2 SO 4Drying over and concentration afforded the crude product of the title compound (4-bromophenyl)-deuterio-methanone (A1), which was used in the next step without further purification.

[0123] N-((4-bromophenyl)methyl-d 2 )-2,2-Dimethoxyethan-1-amine (A2)

[0124] (4-Bromophenyl)-deuterio-methanone (A1) (2.00 g, 10.8 mmol) in toluene (30 mL), p-TsOH H 2 A mixture of 2,2-dimethoxyethan-1-amine (1.05 g, 10.0 mmol) and 2,2-dimethoxyethan-1-amine (0.210 g, 1.08 mmol) was stirred at 138 °C for 3.5 h. The mixture was diluted with saturated NaHCO 3 Pour into aqueous solution (8 mL) of NaHCO 3 The mixture was washed with aqueous solution (2×100 mL) and water (1×100 mL), and the aqueous phase was extracted with toluene (2×100 mL). The combined organic phase was diluted with Na 2 SO 4 The residue was dissolved in EtOH (20 mL) and then diluted with NaBD 4 (220 mg, 5.50 mmol) was added to the mixture at RT and the resulting mixture was stirred at RT for 2 h. Then, HOAc (0.2 mL) was added to the mixture. The reaction mixture was diluted with water and extracted with EtOAc. The extract was washed with brine and Na 2 SO 4 Dry and concentrate to give the title compound N-((4-bromophenyl)methyl-d 2 The crude product of 2,2-dimethoxyethan-1-amine (A2) was obtained. MS-ESI (m / z): 276 [M+1] + .

[0125] N-((4-bromophenyl)methyl-d 2 )-N-(2,2-dimethoxyethyl)-4-methylbenzenesulfonamide (A3)

[0126] N-((4-bromophenyl)methyl-d 2 To a solution of N-((4-bromophenyl)methyl-d)-2,2-dimethoxyethan-1-amine (A2) (2.5 g, 9.1 mmol) in DCM (16 mL) and pyridine (2 mL) was added TsCl (3.10 g, 16.3 mmol). The mixture was stirred at RT for 2 h. The mixture was washed with 1N HCl (3x50 mL), dried and concentrated. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (20:1 to 5:1) to give the title compound N-((4-bromophenyl)methyl-d)- 2 )-N-(2,2-dimethoxyethyl)-4-methylbenzenesulfonamide (A3) was obtained. MS-ESI (m / z): 430 [M+1] + .

[0127] 6-Bromoisoquinoline-1-d (A4)

[0128]

[0128] AlCl in DCM (10 mL) 3 (2.95 g, 22.0 mmol) of N-((4-bromophenyl)methyl-d 2 A solution of 1.9 g (4.4 mmol) of N-(2,2-dimethoxyethyl)-4-methylbenzenesulfonamide (A3) in 10 mL of DCM was added dropwise. After stirring at RT for 4 h, the mixture was poured onto ice (50 g) and extracted with DCM (3×100 mL). The organic phase was washed with 1N NaOH (2×100 mL), NaHCO 3 It was washed successively with aqueous solution (1×100 mL) and brine (100 mL), dried and concentrated. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (5:1) to give the title compound 6-bromoisoquinoline-1-d (A4). MS-ESI (m / z): 209 [M+1] + .

[0129] Isoquinolin-1-d-6-amine (A)

[0130] To a mixture of 6-bromoisoquinoline-1-d (A4) (455 mg, 2.19 mmol), diphenylmethanimine (441 μl, 2.62 mmol) and t-BuONa (631 mg, 6.57 mmol) in toluene (10 mL), BINAP (954 mg, 1.53 mmol) and Pd 2 (dba) 3 (401 mg, 0.438 mmol) was added at RT. The mixture was stirred at 100° C. for 3 h. The mixture was concentrated. The residue was dissolved in MeOH (20 mL) and then NH 2 OH·HCl (241 mg, 3.50 mmol) and NaOAc (312 mg, 3.80 mmol) were added to the mixture at RT, and the resulting mixture was stirred at RT for 1.5 h. The mixture was concentrated, and the residue was purified by silica gel column chromatography eluting with PE / EtOAc (1:1) to give the title compound isoquinolin-1-d-6-amine (A). MS-ESI (m / z): 146 [M+1] + .

[0131]

[0131] Intermediate B Isoquinolin-3-d-6-amine (Ba) and isoquinolin-3,4-d2-6-amine (Bb) [ka]

[0132] 2,2-Dimethoxyethane-1,1-d 2 -1-amine (B1)

[0133]

[0133] A solution of 2,2-dimethoxyacetamide (0.10 g, 0.84 mmol) in THF (3 mL) was diluted with LiAlD 4 (42 mg, 1.0 mmol) was added at RT and the mixture was stirred at 65 °C for 1.5 h. Then, NaSO 4 10H 2 O was added to the mixture under ice-water bath. After stirring for 20 min, the mixture was filtered. The filter cake was washed with THF, and the solution was evaporated to give 2,2-dimethoxyethane-1,1-d 2The crude product of -1-amine (B1) was obtained and used in the next step without further purification. MS-ESI (m / z): 108 [M+1] + .

[0134] N-(4-bromobenzyl)-2,2-dimethoxyethane-1,1-d 2 -1-amine (B2)

[0135] 2,2-Dimethoxyethane-1,1-d in toluene (50 mL) 2 -1-Amine (B1) (100 mg, 0.920 mmol), p-TsOH H 2 A mixture of 2H2O (17.5 mg, 0.0920 mmol) and 4-bromobenzaldehyde (170 mg, 0.920 mmol) was stirred at 135 °C for 4.5 h. After concentration, the residue was dissolved in EtOH (30 mL) and then added NaBH 4 (300 mg, 7.90 mmol) was added to the mixture at RT. The resulting mixture was stirred at RT for 1 h. Then, HOAc (0.2 mL) was added to the mixture. The solvent was concentrated, and the residue was diluted with water and extracted with EtOAc. The extract was washed with brine and Na 2 SO 4 Dry and concentrate to give the title compound N-(4-bromobenzyl)-2,2-dimethoxyethane-1,1-d 2 The crude product of -1-amine (B2) was obtained. MS-ESI (m / z): 276 [M+1] + .

[0136]

[0136] Isoquinoline-3-d-6-amine (Ba) and isoquinoline-3,4-d 2 -6-Amine (Bb)

[0137] The title compound isoquinoline-3-d-6-amine (Ba) (30%) and isoquinoline-3,4-d 2 -6-amine (Bb) (70%) was reacted with N-((4-bromophenyl)methyl-d 2)-2,2-dimethoxyethane-1-amine (A2) to N-(4-bromobenzyl)-2,2-dimethoxyethane-1,1-d 2 Prepared according to the synthesis method of A by replacing with -1-amine (B2). MS-ESI (m / z) Ba: 146 [M+1] + and Bb:147[M+1] +

[0138]

[0138] Intermediate C 2,4-bis(methyl-d 3 )benzoic acid (C) [ka]

[0139]

[0139] 2,4-bis(methyl-d 3 )benzoic acid (C)

[0140] 2,4-Dimethylbenzoic acid (1.0 g, 6.1 mmol) in DMSO-d 6 (10 mL) solution was added t-BuOK (3.40 g, 30.5 mmol). The mixture was then purified by N 2 The mixture was stirred at 120° C. for 3 h under reduced pressure. After cooling to RT, the mixture was diluted with water. The pH was adjusted to 2-3 and extracted with EtOAc (3×). The extract was washed with water, brine, and added with Na 2 SO 4 After drying and concentration, a solid was obtained. This solid was treated twice as described above to give the title compound 2,4-bis(methyl-d 3 ) to give benzoic acid (C).

[0141]

[0141] Intermediate D (4-(2-((tert-butyldimethylsilyl)oxy)ethyl)phenyl)methane-d 2 -Oar (D) [ka]

[0142] Ethyl 4-(2-hydroxyethyl)benzoate (D1)

[0143] 2-(4-Bromophenyl)ethan-1-ol (1.0 g, 5.0 mmol), TEA (0.83 ml, 6.0 mmol) and Pd(dppf)Cl in EtOH (35 mL) and DMSO (22 mL). 2 (365 mg, 0.500 mmol) was stirred at 80° C. under CO atmosphere for 3.5 h. The reaction mixture was cooled to RT and saturated NH 4 The mixture was quenched with aqueous Cl and extracted with EtOAc. The extract was washed with brine (30 mL) and 2 SO 4 The mixture was dried over 100 ml and concentrated. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (5:1 to 4:1) to obtain the title compound ethyl 4-(2-hydroxyethyl)benzoate (D1). MS-ESI (m / z): 195 [M+1] + .

[0144] Ethyl 4-(2-((tert-butyldimethylsilyl)oxy)ethyl)benzoate (D2)

[0145] To a solution of ethyl 4-(2-hydroxyethyl)benzoate (D1) (0.96 g, 4.9 mmol) in DMF (20 mL) was added imidazole (0.84 g, 12 mmol). The mixture was cooled to 0° C., and then TBSCl (0.894 g, 5.91 mmol) was added. The mixture was stirred at RT for 30 min, diluted with water, and extracted with EtOAc. The extract was washed with brine (30 mL) and diluted with Na 2 SO 4 The mixture was dried over 100 ml and concentrated. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (50:1) to give the title compound ethyl 4-(2-((tert-butyldimethylsilyl)oxy)ethyl)benzoate (D2). MS-ESI (m / z): 309 [M+1] + .

[0146] (4-(2-((tert-butyldimethylsilyl)oxy)ethyl)phenyl)methane-d 2 -Oar (D)

[0147] The title compound (4-(2-((tert-butyldimethylsilyl)oxy)ethyl)phenyl)methane-d 2 2,2-Dimethoxyethane-1,1-d-ol (D) can be prepared by replacing 2,2-dimethoxyacetamide with ethyl 4-(2-((tert-butyldimethylsilyl)oxy)ethyl)-benzoate (D2). 2 Prepared according to the synthesis method of -1-amine (B1). MS-ESI (m / z): 269 [M+1] + .

[0148] Intermediate E Methyl 2-(4-(hydroxymethyl-d 2 )Phenyl)acetate-d 2 (E) [ka]

[0149]

[0149] 1,4-phenylenebis(methane-d 2 -All) (E1)

[0150]

[0150] A solution of dimethyl terephthalate (5.9 g, 30 mmol) in THF (100 mL) was added to LiAlD 4 (3.5 g, 59 mmol) was added in portions. The mixture was stirred at 65° C. for 1 h. After cooling to 0° C., the mixture was diluted with Na 2 SO 4 10H 2 The mixture was carefully quenched with O and filtered. The filtrate was concentrated to give the title compound, 1,4-phenylenebis(methane-d 2 -all) (E1).

[0151]

[0151] (4-((((tert-butyldimethylsilyl)oxy)methyl-d 2)Phenyl)methane-d 2 -Oar (E2)

[0152] To a solution of 1,4-phenylenebis(methane-d2-ol) (E1) (4.3 g, 30 mmol) and 1H-imidazole (2.2 g, 33 mmol) in DMF (50 mL) was added tert-butylchlorodimethylsilane (8.3 g, 30 mmol). After stirring at RT for 18 h, the mixture was diluted with H 2 The extracts were poured into H2O and extracted with EtOAc (3x). 2 Wash with NaO, brine, 2 SO 4 Dry and concentrate to give the title compound (4-(((tert-butyldimethylsilyl)oxy)methyl-d 2 )Phenyl)methane-d 2 -ol (E2) was obtained.

[0153] tert-Butyl ((4-(iodomethyl-d 2 )phenyl)methoxy-d 2 )Dimethylsilane (E3)

[0154] (4-(((tert-butyldimethylsilyl)oxy)methyl-d in DCM (60 mL) 2 )Phenyl)methane-d 2 A mixture of 1H-imidazole (E2) (3.8 g, 10 mmol), 1H-imidazole (1.05 g, 15.0 mmol), iodine (3.3 g, 13 mmol) and triphenylphosphane (3.9 g, 15 mmol) was reacted with N 2 The mixture was stirred at RT under atmospheric pressure for 5 h. The mixture was diluted with DCM and 2 S 2 O 3 Wash with aqueous solution and brine, then add Na 2 SO 4 The residue was purified by silica gel column chromatography eluting with PE / EtOAc (10:1) to give the title compound tert-butyl ((4-(iodomethyl-d 2 )phenyl)methoxy-d 2) Dimethylsilane (E3) was obtained.

[0155] 2-(4-(((tert-butyldimethylsilyl)oxy)methyl-d 2 )Phenyl)acetonitrile-d 2 (E4)

[0156] tert-Butyl ((4-(iodomethyl-d 2 )phenyl)methoxy-d 2 ) dimethylsilane (E4) (3.6 g, 7.3 mmol) and K 2 CO 3 (1.8 g, 13 mmol) was added with TMSCN (1.6 mL, 13 mmol). After refluxing for 20 h, the mixture was cooled to RT and filtered. The filtrate was concentrated. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (10:1) to give the title compound 2-(4-(((tert-butyldimethylsilyl)oxy)methyl-d 2 )Phenyl)acetonitrile-d 2 (E5) was obtained.

[0157] 2-(4-(hydroxymethyl-d 2 )Phenyl)acetic acid-2,2-d 2 (E6)

[0158]

[0158] NaH (5.00 g, 120 mmol) D 2 To a solution in 20 mL of 2-(4-(((tert-butyldimethylsilyl)oxy)methyl-d 2 )Phenyl)acetonitrile-d 2 (E5) (2.5 g, 6.0 mmol) was added. The mixture was stirred at 80° C. for 15 h. After cooling to RT, the mixture was 2 The mixture was diluted with 2×O (30 ml), the pH adjusted to 2-3, and extracted with EtOAc (3×). The extracts were washed with brine and 2 SO 4 The solvent was concentrated to give the title compound 2-(4-(hydroxymethyl-d2 )Phenyl)acetic acid-2,2-d 2 (E6) was obtained. MS-ESI (m / z): 169 [M-1] - .

[0159] Methyl 2-(4-(hydroxymethyl)-d 2 )Phenyl)acetate-d 2 (E)

[0160] 2-(4-(hydroxymethyl-d 2 )Phenyl)acetic acid-2,2-d 2 A solution of (E6) (1.35 g, 7.94 mmol) in MeOH (13 mL) was added to concentrated H 2 SO 4 (1.3 mL) was added dropwise. The mixture was stirred at RT overnight. The mixture was poured into water and extracted with EtOAc (3x). The extracts were washed with saturated NaHCO 3 (aqueous) and brine, and then washed with Na 2 SO 4 Dry and concentrate to obtain the title compound methyl 2-(4-(hydroxymethyl-d 2 )Phenyl)acetate-d 2 (E) was obtained. MS-ESI (m / z): 185 [M+1] + .

[0161] Intermediate F tert-Butyl ((1H-benzo[d]1,2,3]triazol-1-yl)methyl-d2)carbamate (F) [ka]

[0162] (1H-benzo[d][1,2,3]triazol-1-yl)methane-d 2 -Oar (F1)

[0163]

[0163] H of 1H-benzo[d][1,2,3]triazole (625 mg, 5.25 mmol) 2 Formaldehyde-d2 (D 2 20% in 0) (840 mg, 5.25 mmol). The mixture was stirred at RT for 3 h. The solid was collected by filtration and dried to give the title compound (1H-benzo[d][1,2,3]triazol-1-yl)methane-d 2 -ol (F1) was obtained. MS-ESI (m / z): 152 [M+1] + .

[0164]

[0164] tert-Butyl ((1H-benzo[d][1,2,3]triazol-1-yl)methyl-d 2 ) Carbamate (F)

[0165] (1H-benzo[d][1,2,3]triazol-1-yl)methane-d in toluene (50 mL) 2 A mixture of tert-butyl((1H-benzo[d][1,2,3]triazol-1-yl)methyl-d-ol (F1) (650 mg, 4.28 mmol), tert-butyl carbamate (500 mg, 4.28 mmol) and 4-methylbenzenesulfonic acid (26.7 mg, 0.155 mmol) was refluxed for 1 h using a Dean-Stark trap. After cooling to RT, the mixture was concentrated to ¼ volume. The solid was then collected by filtration and dried to give the title compound tert-butyl((1H-benzo[d][1,2,3]triazol-1-yl)methyl-d 2 ) Carbamate (F) was obtained. MS-ESI (m / z): 251 [M+1] + .

[0166] Example 1 (S)-4-(3-amino-1-((isoquinolin-6-yl-1-d)amino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (1) [ka]

[0167] (S)-3-((tert-butoxycarbonyl)amino)-2-(4-(((2,4-dimethylbenzoyl)oxy)methyl)phenyl)-propanoic acid (1a)

[0168] (S)-3-((tert-butoxycarbonyl)amino)-2-(4-(((2,4-dimethylbenzoyl)oxy)methyl)phenyl)propanoic acid (1a) was prepared according to the method described in WO 2017 / 086941.

[0169] (S)-4-(3-((tert-butoxycarbonyl)amino)-1-((isoquinolin-6-yl-1-d)amino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (1b)

[0170] To a mixture of (S)-3-((tert-butoxycarbonyl)amino)-2-(4-(((2,4-dimethylbenzoyl)oxy)methyl)phenyl)propanoic acid (1a) (56 mg, 0.13 mmol), isoquinolin-1-d-6-amine (A) (25 mg, 0.17 mmol) and 2,4-lutidine (18 mg, 0.17 mmol) in DMF (1 mL) was added 1,1,1-trichloro-2-methylpropan-2-yl carbonochloridate (41 mg, 0.17 mmol) and stirred at RT for 3 h. The mixture was poured into water (20 mL), extracted with EtOAc (3x50 mL), washed with brine (15 mL) and diluted with Na. 2 SO 4 The residue was purified by preparative TLC eluting with DCM / MeOH (15:1) to give (S)-4-(3-((tert-butoxycarbonyl)amino)-1-((isoquinolin-6-yl-1-d)amino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (1b). MS-ESI (m / z): 555 [M+1] + .

[0171] (S)-4-(3-amino-1-((isoquinolin-6-yl-1-d)amino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (1)

[0172] To a mixture of (S)-4-(3-((tert-butoxycarbonyl)amino)-1-((isoquinolin-6-yl-1-d)-amino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (1b) (25 mg, 0.050 mmol) in DCM (5 mL) was added TFA (1 mL) dropwise. The mixture was stirred at RT for 1 h. The mixture was diluted with saturated NaHCO 3 The organic layer was washed with brine and extracted with DCM (3x30 mL). 2 SO 4 The residue was dissolved in NH in DCM / MeOH and concentrated. 3 Purification by preparative TLC eluting with 1,2-hexanediaminetetraacetate (20:1) gave (S)-4-(3-amino-1-((isoquinolin-6-yl-1-d)amino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (1). MS-ESI (m / z): 455 [M+1] + .

[0173] Example 2 (S)-4-(3-amino-1-((isoquinolin-6-yl-3-d)amino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (2-A) and (S)-4-(3-amino-1-((isoquinolin-6-yl-3,4-d 2 )Amino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (2-B) [ka]

[0174] (S)-4-(3-amino-1-((isoquinolin-6-yl-3-d)amino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (2)

[0175] The title compound (S)-4-(3-amino-1-((isoquinolin-6-yl-3-d)amino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (2-A) (30%) and (S)-4-(3-amino-1-((isoquinolin-6-yl-3,4-d 2 )amino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (2-B) (70%), isoquinolin-1-d-6-amine (A) and isoquinolin-3-d-6-amine (Ba) and isoquinoline-3,4-d 2 -6(Bb) was replaced by (S)-4-(3-amino-1-((isoquinolin-6-yl-1-d)amino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (1) according to the synthesis method. MS-ESI(m / z):2-A:455[M+1] + and 2-B: 456[M+1] +

[0176] Example 3 (S)-(4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenyl)methyl-d 2 2,4-Dimethylbenzoate (3) [ka]

[0177] (4-(2-((tert-butyldimethylsilyl)oxy)ethyl)phenyl)methyl-d 2 2,4-Dimethylbenzoate (3a)

[0178] To a solution of 2,4-dimethylbenzoic acid (31.0 mg, 0.204 mmol) in DCM (1 mL) was added EDCI (65.0 mg, 0.335 mmol). The mixture was stirred at RT for 1 h and then (4-(2-((tert-butyldimethylsilyl)oxy)ethyl)phenyl)methane-d 2A solution of .alpha.-ol (D) (50.0 mg, 0.186 mmol) in DCM and DMAP (5.0 mg, 0.0372 mmol) were added. The mixture was stirred at RT overnight. The mixture was diluted with water and extracted with DCM (3.times.25 mL). The extract was washed with brine (15 mL) and diluted with Na. 2 SO 4 The mixture was dried over and concentrated. The residue was purified by silica gel column chromatography eluting with PE / EtOAc (50:1) to give the title compound (4-(2-((tert-butyldimethylsilyl)oxy)ethyl)phenyl)methyl-d 2 2,4-Dimethylbenzoate (3a) was obtained. MS-ESI (m / z): 401 [M+1] + .

[0179] (4-(2-hydroxyethyl)phenyl)methyl-d 2 2,4-Dimethylbenzoate (3b)

[0180] (4-(2-((tert-butyldimethylsilyl)oxy)ethyl)phenyl)methyl-d 2 To a solution of 2,4-dimethylbenzoate (3a) (4.100 g, 10.25 mmol) in MeOH (20 mL) was added 1N HCl (2 mL). The mixture was stirred at RT for 1 h. The mixture was diluted with water and extracted with DCM (3x25 mL). The extract was washed with brine and diluted with Na 2 SO 4 Dry and concentrate to give the title compound (4-(2-hydroxyethyl)phenyl)methyl-d 2 2,4-Dimethylbenzoate (3b) was obtained. MS-ESI (m / z): 287 [M+1] + .

[0181] (4-(2-oxoethyl)phenyl)methyl-d 2 2,4-Dimethylbenzoate (3c)

[0182] (4-(2-hydroxyethyl)phenyl)methyl-d 2To a solution of 2,4-dimethylbenzoate (3b) (2.90 g, 10.3 mmol) in DCM (100 mL) was added DMP (5.20 g, 12.3 mmol). The mixture was stirred at RT for 30 min. The reaction was diluted with saturated NaHCO 3 The mixture was quenched with aqueous solution (100 mL) and extracted with DCM. The organic phase was washed with brine (20 mL) and added Na 2 SO 4 The residue was purified by silica gel column chromatography eluting with PE / EtOAc (10:1) to give the title compound (4-(2-oxoethyl)phenyl)methyl-d 2 2.4-Dimethylbenzoate (3c) was obtained. MS-ESI (m / z): 285 [M+1] + .

[0183] 2-(4-(((2,4-dimethylbenzoyl)oxy)methyl-d 2 )Phenyl)acetic acid (3d)

[0184] (4-(2-oxoethyl)phenyl)methyl-d 2 To a solution of 2,4-dimethylbenzoate (3c) (50 mg, 0.18 mmol) in DMF (1 mL) was added oxone (36 mg, 0.21 mmol). The mixture was stirred at RT for 4 h. The mixture was poured into water (20 mL). The precipitated solid was collected by filtration, washed with water (30 mL), dried and purified to give 2-(4-(((2,4-dimethylbenzoyl)oxy)-methyl-d 2 The crude product of 3d)phenyl)acetic acid was obtained and used in the next step without further purification. MS-ESI (m / z): 301 [M+1] + .

[0185] (4-(2-chloro-2-oxoethyl)phenyl)methyl-d 2 2,4-Dimethylbenzoate (3e)

[0186] 2-(4-(((2,4-dimethylbenzoyl)oxy)-methyl-d 2To a solution of (4-(2-chloro-2-oxoethyl)phenyl)acetic acid (3d) (1.9 g, 6.2 mmol) in DCM (40 mL) was added oxalyl chloride (0.79 ml, 7.4 mmol). The mixture was stirred at RT overnight. The mixture was concentrated to give (4-(2-chloro-2-oxoethyl)phenyl)methyl-d 2 The crude product of 2,4-dimethylbenzoate (3e) was obtained and used in the next step without further purification.

[0187]

[0187] (R)-(4-(2-(4-benzoyl-2-oxooxazolidin-3-yl)-2-oxoethyl)phenyl)methyl-d 2 2,4-Dimethylbenzoate (3f)

[0188] To a solution of (R)-4-benzyloxazolidin-2-one (1.1 g, 6.2 mmol) in THF (40 mL) was added n-BuLi (1.6 M, 4.3 mL, 6.8 mmol) dropwise at −70° C., followed by the addition of (4-(2-chloro-2-oxoethyl)phenyl)methyl-d 2 A solution of 2,4-dimethylbenzoate (3e) (2.0 g, 6.2 mmol) in THF was added dropwise and the resulting mixture was stirred at −70° C. for 1 h. The reaction was washed with saturated NH 4 The mixture was quenched with aqueous Cl and extracted with EtOAc (2x50 mL). The extracts were washed with brine and 2 SO 4 The residue was purified by silica gel column chromatography eluting with PE / EtOAc (10:1 to 5:1) to give the title compound (R)-(4-(2-(4-benzyl-2-oxooxazolidin-3-yl)-2-oxoethyl)phenyl)methyl-d 2 2,4-Dimethyl-benzoate (3f) was obtained. MS-ESI (m / z): 460 [M+1] + .

[0189]

[0189] (4-((S)-1-((R)-4-benzyl-2-oxooxazolidin-3-yl)-3-((tert-butoxycarbonyl)amino)-1-oxopropan-2-yl)phenyl)methyl-d 22,4-Dimethylbenzoate (3g)

[0190] (R)-(4-(2-(4-benzyl-2-oxooxazolidin-3-yl)-2-oxoethyl)phenyl)-methyl-d 2 To a solution of 2,4-dimethylbenzoate (3f) (1.0 g, 2.2 mmol) in THF (20 mL) was added dropwise KHMDS (1.0 M, 2.6 mL, 2.61 mmol) at -70°C. The mixture was stirred at -70°C for 30 min, then a solution of tert-butyl ((1H-benzo[d][1,2,3]triazol-1-yl)methyl)carbamate (0.65 g, 2.6 mmol) (WO 2017 / 086941) in THF (3 mL) was added dropwise at -70°C. The mixture was stirred at 0°C for 1 h. The reaction was purified with saturated NH 4 The mixture was quenched with aqueous Cl and extracted with EtOAc (3x50 mL). The extracts were washed with brine and 2 SO 4 The residue was purified by silica gel column chromatography eluting with PE / EtOAc (10:1 to 5:1) to give the title compound (4-((S)-1-((R)-4-benzyl-2-oxooxazolidin-3-yl)-3-((tert-butoxycarbonyl)amino)-1-oxopropan-2-yl)phenyl)methyl-d 2 2,4-Dimethylbenzoate (3g) was obtained. MS-ESI(m / z): 589[M+1] + .

[0191] (S)-3-((tert-butoxycarbonyl)amino)-2-(4-(((2,4-dimethylbenzoyl)oxy)methyl-d 2 )Phenyl)propanoic acid (3h)

[0192]

[0192] (4-((S)-1-((R)-4-benzyl-2-oxooxazolidin-3-yl)-3-((tert-butoxycarbonyl)amino)-1-oxopropan-2-yl)phenyl)methyl-d 2 2,4-Dimethylbenzoate (3g) (1.1 g, 1.9 mmol) in THF / H2 HO (22 ml / 7 mL) solution at 0°C. 2 O 2 (30%, 0.85 g, 7.5 mmol) and LiOH H 2 O (78 mg, 1.9 mmol) was added sequentially. The mixture was stirred at 0° C. for 10 min. The reaction was diluted with 10% Na 2 SO 3 The mixture was quenched with aqueous EtOAc (3×50 mL). The mixture was extracted with EtOAc (3×50 mL). The extract was washed with brine and 2 SO 4 The residue was purified by silica gel column chromatography eluting with DCM / MeOH (100:1 to 20:1) to give the title compound (S)-3-((tert-butoxycarbonyl)amino)-2-(4-(((2,4-dimethylbenzoyl)oxy)methyl-d 2 )phenyl)propanoic acid (3h). MS-ESI (m / z): 430 [M+1] + .

[0193] (S)-(4-(3-((tert-butoxycarbonyl)amino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenyl)methyl-d 2 2,4-Dimethylbenzoate (3i)

[0194] Isoquinolin-6-amine (23 mg, 0.16 mmol) and (S)-3-((tert-butoxycarbonyl)amino)-2-(4-(((2,4-dimethylbenzoyl)oxy)methyl-d 2 To a solution of phenyl(propanoic acid (3h) (43 mg, 0.10 mmol) in DMF (1 mL) was added EDCI (39 mg, 0.21 mmol), TEA (33 mg, 0.30 mmol) and HOBT (27 mg, 0.21 mmol). The mixture was stirred at RT overnight. The mixture was diluted with water and extracted with EtOAc (3x5 mL). The extract was washed with brine and diluted with Na 2 SO 4The residue was purified by silica gel column chromatography eluting with DCM / MeOH (15:1) to give the title compound (S)-(4-(3-((tert-butoxycarbonyl)amino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenyl)methyl-d 2 2,4-Dimethylbenzoate (3i) was obtained. MS-ESI (m / z): 556 [M+1] + .

[0195] (S)-(4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenyl)methyl-d 2 2,4-Dimethylbenzoate (3)

[0196] The title compound (S)-(4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenyl)methyl-d 2 2,4-Dimethylbenzoate (3) was converted to (S)-4-(3-((tert-butoxycarbonyl)amino)-1-((isoquinolin-6-yl-1-d)amino)-1-oxopropan-2-yl)benzyl 2,4-Dimethylbenzoate (1b) was converted to (S)-(4-(3-((tert-butoxycarbonyl)amino)-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)phenyl)methyl-d 2 Prepared according to the synthesis method of (S)-4-(3-amino-1-((isoquinolin-6-yl-1-d)amino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (1) by replacing with 2,4-dimethylbenzoate (3i). MS-ESI(m / z): 456[M+1] + .

[0197] Example 4 (S)-4-(3-amino-1-((isoquinolin-6-yl-1-d)amino)-1-oxopropan-2-yl-3,3-d 2 ) Benzyl 2,4-dimethylbenzoate dihydrochloride (4) [ka]

[0198] (R)-4-(2-(4-benzyl-2-oxooxazolidin-3-yl)-2-oxoethyl)benzyl 2,4-dimethylbenzoate (4a)

[0199] The title compound (R)-4-(2-(4-benzyl-2-oxooxazolidin-3-yl)-2-oxoethyl)benzyl 2,4-dimethylbenzoate (4a) was prepared according to the method described in WO 2017 / 086941. MS-ESI (m / z): 458 [M+1] + .

[0200]

[0200] 4-((S)-1-((R)-4-benzyl-2-oxooxazolidin-3-yl)-3-((tert-butoxycarbonyl)amino)-1-oxopropan-2-yl-3,3-d 2 ) 2,4-Dimethylbenzoate (4b)

[0201] To a solution of the title compound (R)-4-(2-(4-benzyl-2-oxooxazolidin-3-yl)-2-oxoethyl)benzyl 2,4-dimethylbenzoate (4a) (1.0 g, 2.2 mmol) in THF (20 mL) was added dropwise KHMDS (1.0 M, 2.6 mL, 2.61 mmol) at −70° C. The mixture was stirred at −70° C. for 30 minutes, and then tert-butyl((1H-benzo[d][1,2,3]triazol-1-yl)methyl-d 2 A solution of carbamate (F) (0.65 g, 2.6 mmol) in THF (3 mL) was added dropwise at -70 °C. The mixture was stirred at 0 °C for 1 h. The reaction was diluted with saturated NH 4 The mixture was quenched with aqueous Cl and extracted with EtOAc (3x50 mL). The extracts were washed with brine and 2 SO 4The residue was purified by silica gel column chromatography eluting with PE / EtOAc (10:1 to 5:1) to obtain the title compound 4-((S)-1-((R)-4-benzyl-2-oxooxazolidin-3-yl)-3-((tert-butoxycarbonyl)amino)-1-oxopropan-2-yl-3,3-d 2 ) Benzyl 2,4-dimethylbenzoate (4b) was obtained. MS-ESI (m / z): 589 [M+1] + .

[0202]

[0202] (S)-3-((tert-butoxycarbonyl)amino)-2-(4-(((2,4-dimethylbenzoyl)oxy)methyl)phenyl)propane-3,3-d 2 acid(4c)

[0203]

[0203] The title compound (S)-3-((tert-butoxycarbonyl)amino)-2-(4-(((2,4-dimethylbenzoyl)oxy)methyl)phenyl)propane-3,3-d 2 Acid 4c was reacted with (4-((S)-1-((R)-4-benzyl-2-oxooxazolidin-3-yl)-3-((tert-butoxycarbonyl)amino)-1-oxopropan-2-yl)phenyl)methyl-d 2 2,4-Dimethylbenzoate (3g) to 4-((S)-1-((R)-4-benzyl-2-oxooxazolidin-3-yl)-3-((tert-butoxycarbonyl)amino)-1-oxopropan-2-yl-3,3-d 2 ) benzyl 2,4-dimethylbenzoate (4b) to give (S)-3-((tert-butoxycarbonyl)amino)-2-(4-(((2,4-dimethylbenzoyl)oxy)methyl-d 2 Prepared according to the synthesis method of 3h. MS-ESI (m / z): 430 [M+1] +

[0204] (S)-4-(3-amino-1-(((isoquinolin-6-yl-1-d)amino)-1-oxopropan-2-yl-3,3-d 2 ) Benzyl 2,4-dimethylbenzoate (4d)

[0205] The title compound (S)-4-(3-amino-1-((isoquinolin-6-yl-1-d)amino)-1-oxopropan-2-yl-3,3-d 2 ) benzyl 2,4-dimethylbenzoate (4d), (S)-3-((tert-butoxycarbonyl)amino)-2-(4-(((2,4-dimethylbenzoyl)oxy)methyl)phenyl)propanoic acid (1a) and (S)-3-((tert-butoxycarbonyl)amino)-2-(4-(((2,4-dimethylbenzoyl)oxy)methyl)phenyl)propane-3,3-d 2 Prepared according to the synthesis method of (S)-4-(3-amino-1-((isoquinolin-6-yl-1-d)amino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate (1) by replacing with acid (4c). MS-ESI (m / z): 457 [M+1] + .

[0206] (S)-4-(3-amino-1-((isoquinolin-6-yl-1-d)amino)-1-oxopropan-2-yl-3,3-d 2 ) Benzyl 2,4-dimethylbenzoate dihydrochloride (4)

[0207] (S)-4-(3-amino-1-((isoquinolin-6-yl-1-d)amino)-1-oxopropan-2-yl-3,3-d 2 To a solution of benzyl 2,4-dimethylbenzoate (4d) (22 mg, 0.048 mmol) in DCM (1 mL) and MeOH (0.1 mL) was added HCl (30% in EtOH, 0.108 mmol). After stirring for 10 min, the mixture was concentrated to give the title compound (S)-4-(3-amino-1-((isoquinolin-6-yl-1-d)amino)-1-oxopropan-2-yl-3,3-d 2) Benzyl 2,4-dimethylbenzoate dihydrochloride (4) was obtained. MS-ESI (m / z): 457 [M+1] + .

[0208] Example 5 (S)-(4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl-2,3,3-d 3 )phenyl)methyl-d 2 2,4-bis(methyl-d 3 ) Benzoate dihydrochloride (5) [ka]

[0209] (4-(2-Methoxy-2-oxoethyl-1,1-d2)phenyl)methyl-d 2 2,4-bis(methyl-d 3 ) Benzoate (5a)

[0210] Methyl 2-(4-hydroxymethyl-d 2 )Phenyl)acetate-d 2 (E) (610 mg, 3.30 mmol), 2,4-bis(methyl-d 3 A mixture of benzoic acid (C) (517 mg, 3.30 mmol), EDCI (1.2 g, 6.0 mmol) and DMAP (81 mg, 0.66 mmol) was stirred at RT overnight. The mixture was diluted with DCM and 2 Wash with NaCl and brine. 2 SO 4 The residue was purified by silica gel column chromatography eluting with PE / EtOAc (10:1) to give the title compound (4-(2-methoxy-2-oxoethyl-1,1-d 2 )phenyl)methyl-d 2 2,4-bis(methyl-d 3 ) benzoate (5a). MS-ESI (m / z): 323 [M+1] + .

[0211]

[0211] 2-(4-(((2,4-bis(methyl-d 3 )benzoyl)oxy)methyl-d 2 )Phenyl)acetic acid-2,2-d 2 (5b)

[0212]

[0212] (4-(2-methoxy-2-oxoethyl-1,1-d 2 )phenyl)methyl-d 2 2,4-bis(methyl-d 3 ) Benzoate (5a) (752 mg, 2.33 mmol) D 2 To a solution of LiOH.H2O (15 mL) and THF (10 mL) 2 HO (108 mg, 2.6 mmol) was added. The mixture was stirred at RT for 2 h. The mixture was acidified to pH = 2-3 and diluted with H 2 The mixture was diluted with O and extracted with EtOAc (3x). The combined EtOAc phase was washed with H 2 Wash with NaCl and brine. 2 SO 4 Dry and concentrate to give the title compound 2-(4-(((2,4-bis(methyl-d 3 )benzoyl)oxy)methyl-d 2 )Phenyl)acetic acid-2,2-d 2 (5b) was obtained. MS-ESI (m / z): 307 [M-1] - .

[0213] (R)-(4-(2-(4-benzyl-2-oxooxazolidin-3-yl)-2-oxoethyl-1,1-d 2 )phenyl)methyl-d 2 2,4-bis(methyl-d 3 ) Benzoate (5c)

[0214] The title compound (R)-(4-(2-(4-benzyl-2-oxooxazolidin-3-yl)-2-oxoethyl-1,1-d 2 )phenyl)methyl-d 2 2,4-bis(methyl-d 3) benzoate (5c) to 2-(4-(((2,4-dimethylbenzoyl)oxy)-methyl-d 2 )phenyl)acetic acid (3d) to 2-(4-(((2,4-bis(methyl-d 3 )benzoyl)oxy)methyl-d 2 )Phenyl)acetic acid-2,2-d 2 Prepared according to the synthetic method of 3f by replacing with (5b). MS-ESI (m / z): 468 [M+1] + .

[0215] (S)-(4-(1-((R)-4-benzyl-2-oxooxazolidin-3-yl)-3-((tert-butoxycarbonyl)amino)-1-oxopropan-2-yl-2,3,3-d 3 )phenyl)methyl-d 2 2,4-bis(methyl-d 3 ) Benzoate (5d)

[0216] The title compound (S)-(4-(1-((R)-4-benzyl-2-oxooxazolidin-3-yl)-3-((tert-butoxycarbonyl)amino)-1-oxopropan-2-yl-2,3,3-d 3 ) Phenyl(methyl-d 2 2,4-bis(methyl-d 3 ) benzoate (5d) was converted to (R)-(4-(2-(4-benzyl-2-oxooxazolidin-3-yl)-2-oxoethyl)phenyl)-methyl-d 2 2,4-Dimethylbenzoate (3f) and tert-butyl ((1H-benzo[d][1,2,3]triazol-1-yl)methyl)carbamate were reacted with (R)-(4-(2-(4-benzyl-2-oxooxazolidin-3-yl)-2-oxoethyl-1,1-d 2 )phenyl)methyl-d 2 2,4-bis(methyl-d 3 ) benzoate (5c) and tert-butyl((1H-benzo[d][1,2,3]triazol-1-yl)methyl-d 2Prepared according to the synthesis method of 3g by replacing with carbamate (F). MS-ESI (m / z): 598 [M+1] + .

[0217]

[0217] (S)-2-(4-(((2,4-bis(methyl-d 3 )benzoyl)oxy)methyl-d2)phenyl)-3-((tert-butoxycarbonyl)amino)propane-2,3,3-d 3 Acid (5e)

[0218] (S)-(4-(1-((R)-4-benzyl-2-oxooxazolidin-3-yl)-3-((tert-butoxycarbonyl)amino)-1-oxopropan-2-yl-2,3,3-d 3 )phenyl)methyl-d 2 2,4-bis(methyl-d 3 ) Benzoate (5d) (538 mg, 0.904 mmol) D 2 To a solution in 20O (5 mL) and THF (10 mL) was added H 2 O 2 (0.41 ml, 3.61 mmol), followed by LiOH.H 2 O (42 mg, 0.99 mmol) was added. The mixture was stirred at 0° C. for 0.5 h and then NaHSO 3 (10% aqueous solution). The mixture was acidified to pH=2-3 and extracted with EtOAc (3x50 mL). The extracts were washed with brine and 2 SCO 4 The mixture was dried over 100 ml and concentrated. The residue was purified by silica gel column chromatography eluting with DCM / MeOH (100:1 to 10:1) to give the title compound (S)-2-(4-(((2,4-bis(methyl-d 3 )benzoyl)oxy)-methyl-d 2 )phenyl)-3-((tert-butoxycarbonyl)amino)propane-2,3,3-d 3 Acid (5e) was obtained. MS-ESI (m / z): 439 [M-1] - .

[0219] (S)-(4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl-2,3,3-d 3 )phenyl)methyl-d 2 2,4-bis(methyl-d 3 ) Benzoate (5f)

[0220] The title compound (S)-(4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl-2,3,3-d 3 )phenyl)methyl-d 2 2,4-bis(methyl-d 3 ) benzoate (5f) was converted to (S)-3-((tert-butoxycarbonyl)amino)-2-(4-(((2,4-dimethylbenzoyl)oxy)methyl-d 2 (S)-2-(4-(((2,4-bis(methyl-d 3 )benzoyl)oxy)methyl-d 2 )phenyl)-3-((tert-butoxycarbonyl)amino)propane-2,3,3-d 3 Prepared according to the synthesis method of 3 by substituting the acid (5e). MS-ESI (m / z): 465 [M+1] + .

[0221] (S)-(4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl-2,3,3-d3)phenyl)methyl-d 2 2,4-bis(methyl-d 3 ) Benzoate dihydrochloride (5)

[0222] (S)-(4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl-2,3,3-d 3 )phenyl)methyl-d 2 2,4-bis(methyl-d 3To a solution of benzoate (5f) (20 mg, 0.043 mmol) in DCM (1 mL) and MeOH (0.1 mL) was added HCl (30% in EtOH, 0.108 mmol). After stirring for 10 min, the mixture was concentrated to give the title compound (S)-(4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl-2,3,3-d 3 )phenyl)methyl-d 2 2,4-bis(methyl-d 3 ) to give the benzoate dihydrochloride (5). MS-ESI (m / z): 465 [M+1] + .

[0223] Example 6 (S)-(4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl-2-d)phenyl)methyl-d 2 2,4-bis(methyl-d 3 ) Benzoate dihydrochloride (6) [ka]

[0224] The title compound (S)-(4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl-2-d)phenyl)methyl-d 2 2,4-bis(methyl-d 3 ) benzoate dihydrochloride (6) was reacted with tert-butyl((1H-benzo[d][1,2,3]triazol-1-yl)methyl-d 2 Prepared according to the synthesis method of 5 by replacing tert-butyl ((1H-benzo[d][1,2,3]triazol-1-yl)methyl)carbamate (F) with tert-butyl ((1H-benzo[d][1,2,3]triazol-1-yl)methyl)carbamate. MS-ESI (m / z): 463 [M+1] + .

[0225]

[0225] Reference compound 1 (S)-4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate dihydrochloride (Reference Compound 1)

[0226] (S)-4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate has been disclosed and was prepared essentially according to the same procedure outlined on page 21 of WO 2017 / 086941. Reference compound 1 was prepared by the procedure outlined on page 21 of WO 2017 / 086941, as follows: (S)-(4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl-2,3,3-dimethylbenzoate 3 )phenyl)methyl-d 2 2,4-bis(methyl-d 3 This compound was prepared according to the synthetic method of (5) by replacing (S)-4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate with (S)-4-(3-amino-1-(isoquinolin-6-ylamino)-1-oxopropan-2-yl)benzyl 2,4-dimethylbenzoate.

[0227] Following essentially the same procedures as described for Examples 1-6, Examples 7-24 listed in Table 1 were prepared from appropriate starting materials that were commercially available or known in the literature. The structures and names of Examples 7-24 are shown in Table 1. [Table 1] JPEG0007652865000030.jpg159149

[0228] Cell proliferation assay ROCK-I(h) in vitro inhibitory activity - Method A ROCK-I(h) kinase assays were performed using the KinaseProfiler™ service of Eurofins Pharma Discovery Services UK Limited. ROCK-I(h) was incubated in 8 mM MOPS pH 7.0 (VWR catalogue no. 441644J), 0.2 mM EDTA (Sigma catalogue no. E5134), 30 μM KEAKEKRQEQIAKRRRLSSLRASTSKSGGSQK (Pepceuticals), 10 mM magnesium acetate (Merck Millipore catalogue no. 105819) and [γ- 33 The wells were incubated with test compounds in assay buffer containing 100% DMSO (Calbiochem catalog number 317275) and 100% DMSO (Calbiochem catalog number 317275) prior to spotting into 96-well assay plates using a Mosquito XI (TTP Labtech) before adding the reaction mix. The reaction was started by adding Mg / ATP mix using a Matrix Wellmate. After 40 minutes of incubation at room temperature, the reaction was stopped by adding 3% phosphoric acid solution (Fisher Scientific catalog number O / 0500 / PB17) using a Matrix Wellmate. An aliquot of the reaction was then spotted onto a filtermat, washed with phosphoric acid, followed by rinsing with methanol, before drying and scintillation counting in a Trilux Wallac Microbeta detector (Perkin Elmer). Results were expressed relative to a control containing only DMSO instead of the test compound. Where applicable, IC50 curve analysis was performed using XLFit version 5.3 (ID Business Solutions). Nonlinear regression analysis is used to fit sigmoidal dose-response (variable slope) curves.

[0229]

[0229] The ROCK-II(h) kinase assay was performed using the KinaseProfiler™ service of Eurofins Pharma Discovery Services UK Limited. ROCK-II(h) was incubated in 50 mM Tris pH 7.5 (VWR Catalog No. 103157P), 0.1 mM EGTA (VWR Catalog No. 20308), 30 μM KEAKEKRQEQIAKRRRLSSLRASTSKSGGSQK (Pepceuticals), 10 mM magnesium acetate and [γ- 33 96-well assay plates were first spotted with test compounds in 100% DMSO (Calbiochem catalogue no. 317275) using a Mosquito XI (TTP Labtech) before adding the reaction mix. The reaction was started by adding the Mg / ATP mix using a Matrix Wellmate. After 40 minutes of incubation at room temperature, the reaction was stopped by adding a 3% phosphoric acid solution (Fisher Scientific catalogue no. O / 0500 / PB17) using a Matrix Wellmate. An aliquot of the reaction was then spotted onto a filtermat, washed with phosphoric acid, followed by rinsing with methanol, before drying and scintillation counting in a Trilux Wallac Microbeta detector (Perkin Elmer). Results were expressed relative to a control containing only DMSO instead of the test compound. Where applicable, IC was calculated using XLFit version 5.3 (ID Business Solutions). 50 Curve analysis was performed: nonlinear regression analysis was used to fit sigmoidal dose-response (variable slope) curves.

[0230]

[0230] Selected compounds prepared as above were assayed according to the biological procedures described in Method A. The results are shown in Table 2. [Table 2]

[0231] ROCK-I(h) In Vitro Inhibitory Activity - Method B ROCK-I (Invitrogen, PV3691) kinase assay was performed using CISBIO's HTRF® KinEASE-STK kit service. Transfer 10nl of compound dilution to each well of the assay plate (784075, Greiner) using an Echo 550. Centrifuge the compound plate at 1000g for 1 minute. Seal the assay plate. Prepare 5X ROCK-I (Invitrogen, PV 3691) in 1x kinase buffer (1x volume of enzyme buffer 5X and 4x volumes of distilled water; 5mM MgCl 2 ; 1 mM DTT). Add 2 μl of 5X ROCK-I to a 384-well assay plate (784075, Greiner). Add 4 μl of 1x kinase buffer to each well of the assay plate and centrifuge the plate at 1000g for 30 sec and RT for 10 min. Prepare 5x TK-substrate-biotin (5 μM) in kinase buffer and 5x ATP (100 μM) in kinase buffer. Start the reaction by adding 2 μl STK-substrate-biotin and 2 μl ATP. Centrifuge the plate at 1000g for 30 sec. Seal the assay plate at RT for 20 min. Prepare 4X Sa-XL 665 (250 nM) and TK-antibody-cryptate in HTRF detection buffer. Add 5 μl Sa-XL 665 and 5 μl TK-antibody-cryptate to each well of the assay plate. Plates are centrifuged at 1000 g for 30 s at RT for 1 h. Fluorescence signals are read at 615 nm (Cryptate) and 665 nm (XL665) on an Envision 2104 plate reader (Perkin Elmer). The ratio (665 / 615 nm) is calculated for each well. IC values ​​are calculated by fitting the % inhibition values ​​and the logarithm of compound concentrations to a nonlinear regression (dose response-variable slope) using GraphPad 6.0. 50 Calculate.

[0232]

[0232] Selected compounds prepared as above were assayed according to the biological procedures described in Method B. The results are shown in Table 3. [Table 3]

[0233] ROCK In Vivo Inhibitory Activity - Method C Oxybuprocaine (Santen Co. JP lot: B2031); carbonate suppressant (Harvard apparatus); TONOVET Plus (Finland icare); Chinchilla rabbits: 2-8 months old and weighing 2.23-4.41 kg (Dongfang Breeding Co., Ltd., Pizhou, License: SCXK(su)2014-0005, Approval Number: 201814493, 201826314).

[0234] Eighteen male rabbits were randomly assigned to three groups (6 rabbits / group): Vehicle control, Rhopressa @ (0.02% netarsudil, equivalent to 0.2mg / ml netarsudil) or 0.2mg / mL of Example 11 was topically administered once a day for 3 days to the right eye (30μL / eye / administration), and the other left eye was treated with saline as an internal control. Clinical observations and ophthalmic examinations were performed once a day. IOP was measured by TONOVET Plus before the first administration, and 0.5, 2, 4, 6 and 24 hours after the first administration on the first and third days.

[0235]

[0235] The following formula was used to calculate the % change in IOP: % change in IOP (ΔIOP%)=[IOP(time)-IOP(control)] / IOP(control)*100).

[0236]

[0236] Selected compounds prepared as above were assayed according to the biological procedures described in Method C. The results of the 24 hour dosing on day 3 are shown in Table 4. [Table 4]

Claims

【Request 1】 【Chemistry 1】 or a pharma- ceutically acceptable salt thereof.

2. The compound is 【Chemistry 2】 2. The compound of claim 1, selected from:

3. The compound is 【Chemistry 3】 2. The compound of claim 1, selected from:

4. The compound or a pharma- ceutically acceptable salt thereof according to any one of claims 1 to 3, wherein the pharma- ceutically acceptable salt is a hydrochloride salt.

5. 5. The compound according to claim 4, or a pharma- ceutically acceptable salt thereof, wherein the pharma- ceutically acceptable salt is a dihydrochloride salt.

6. A pharmaceutical composition comprising a compound according to any one of claims 1 to 5 or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable carrier.

7. 13. Use of a compound according to any one of claims 1 to 5, or a pharma- ceutically acceptable salt thereof, in the preparation of a medicament for treating elevated intraocular pressure.

8. A pharmaceutical composition for use in treating ocular hypertension comprising a compound according to any one of claims 1 to 5 or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable carrier.

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