Plasma kallikrein inhibitors

Abstract

The present invention provides compounds of Formula (I) and pharmaceutical compositions comprising one or more of the compounds, and methods of using the compounds to treat or prevent one or more conditions that can benefit from the inhibition of plasma kallikrein, including hereditary angioedema, uveitis, posterior uveitis, wet age-related macular edema, diabetic macular edema, diabetic retinopathy, and retinal vein occlusion. The compounds are selective inhibitors of plasma kallikrein.

Description

Background of the Invention

[0002] Plasma kallikrein is the zymogen of a trypsin-like serine protease and is present in plasma. Its gene structure is similar to that of factor XI. Overall, the amino acid sequence of plasma kallikrein shares 58% homology with factor XI. Proteolytic activation of factor XI at the internal I389-R390 bond produces a heavy chain (371 amino acids) and a light chain (248 amino acids). The active site of plasma kallikrein is contained within the light chain. The light chain of plasma kallikrein reacts with protease inhibitors, including α2-macroglobulin and Cl- inhibitors. Interestingly, in the presence of high molecular weight kininogen (HMWK), heparin significantly accelerates the inhibition of plasma kallikrein by antithrombin III. In the blood, most plasma kallikrein circulates in a complex with HMWK. Plasma kallikrein cleaves HMWK to release bradykinin. Bradykinin release leads to increased vascular permeability and vasodilation (for review, Coleman, R., “Contact Activation Pathway”, Hemostasis and Thrombosis, pp. 103-122, Lippincott Williams & Wilkins (2001); Schmaier AH, “Contact Activation”, Thrombosis and Hemorrhage, pp. 105-128 (1998)).

[0003] Patients exhibiting a genetic defect on C1-esterase inhibitors have hereditary angioedema (HAE), a lifelong condition that causes intermittent swelling throughout the body, including the hands, feet, face, throat, genitals, and gastrointestinal tract. Analysis of blisters resulting from acute attacks has confirmed the presence of high levels of plasma kallikrein, and treatment with the protein-based, reversible plasma kallikrein inhibitor ikalatide (Kalbitor) has been approved by the FDA for the treatment of acute attacks of HAE (Schneider, L et al., J. Allergy Clin. Immunol., 120: p. 416 (2007)).

[0004] Furthermore, the plasma kallikrein-kinin system is abnormally abundant in patients diagnosed with advanced diabetic macular edema (DME). Recent publications have confirmed that plasma kallikrein promotes retinal vascular leakage and dysfunction observed in diabetic rodent models (A. Clermont et al., Diabetes, 60:1590 (2011)), and treatment with small molecule plasma kallikrein inhibitors improved the observed retinal vascular permeability and other abnormalities related to retinal blood flow.

[0005] There is a need in this field to develop plasma kallikrein inhibitors that can be used to treat a wide variety of disorders, including hereditary angioedema, diabetic macular edema, and diabetic retinopathy. Invention Overview

[0007] This invention relates to compounds of formula I:

[0008]

[0009] Compounds of Formula I and their pharmaceutically acceptable salts. Compounds of Formula I are inhibitors of plasma kallikrein and therefore can be used to treat, inhibit, or improve one or more conditions that can benefit from inhibition of plasma kallikrein, including hereditary angioedema, uveitis, posterior uveitis, wet age-related macular edema, diabetic macular edema, diabetic retinopathy, and retinal vein occlusion. The compounds of the present invention can also be used in combination with other therapeutically effective agents, including, but not limited to, other drugs that can be used to treat hereditary angioedema, uveitis, posterior uveitis, wet age-related macular edema, diabetic macular edema, diabetic retinopathy, and retinal vein occlusion. Furthermore, the present invention relates to methods for preparing compounds of Formula I, and pharmaceutical compositions comprising compounds of Formula I and their pharmaceutically acceptable salts. Invention Details

[0011] This invention relates to compounds of formula I or pharmaceutically acceptable salts thereof:

[0012]

[0013] Where X is N or CH;

[0014] R 1 Selected from hydrogen, halogen, hydroxyl and C 1-6 alkyl;

[0015] R 2 Selected from hydrogen, halogen, hydroxyl and C 1-6 alkyl;

[0016] R 3 Selected from hydrogen, halogen, hydroxyl and C 1-6 Alkyl group, wherein the alkyl group is optionally substituted with 1 to 4 substituents, the substituents being independently selected from halogens, cyano groups, and OR groups. x ;

[0017] R 4 Selected from hydrogen, halogen, hydroxyl and C 1-6 Alkyl; wherein the alkyl group is optionally substituted with 1-4 substituents, the substituents being independently selected from halogens, cyano groups, and OR. x;

[0018] R 5 It is NR 9 R 10 OR x ;

[0019] Each R 6 Independently selected from hydrogen, halogen, hydroxyl and C 1-6 Alkyl group, wherein the alkyl group is optionally substituted with 1-3 halogens;

[0020] Each R 7 Selected from hydrogen, halogen, hydroxyl and C 1-6 Alkyl group, wherein the alkyl group is optionally substituted with 1-3 halogens;

[0021] Or R 6 and R 7 They can form 3-6 membered cycloalkyl groups together with the carbon atoms to which they are attached, wherein the cycloalkyl groups are optionally substituted with one or two halogens;

[0022] R 8 Selected from hydrogen; halogen; hydroxyl group; R x OR x ; Phenylacetylene; Indene; OR y It can be a monocyclic or bicyclic heteroaryl group; a heterocyclic group; and a C group that can be a monocyclic or bicyclic group. 3-6 Cycloalkyl; wherein the phenyl and heteroaryl groups are optionally substituted with 1-3 substituents, the substituents being independently selected from oxo, halogen, and R-type compounds. x OR x NR 9 R 10 NR 9 (C=O)R x NR 9 (C=O)OR x (C=O)OR x (C=O)NR 9 R y and OR y The cycloalkyl and heterocyclic groups are optionally substituted with 1-3 substituents, which are independently selected from oxo, halogen, and R groups. x and OR x ;

[0023] R 9 Is it hydrogen or C? 1-3 alkyl;

[0024] R 10 Is it hydrogen or C? 1-3 alkyl;

[0025] R x Is it hydrogen or C?1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with 1-3 substituents selected from halogens and hydroxyl groups.

[0026] R y It is phenyl, heterocyclic or C 3-6 Cycloalkyl groups, wherein the phenyl group is optionally substituted with 1-3 halogens, the heterocyclic group is optionally substituted with 1 or 2 oxygen groups, and the cycloalkyl group is optionally C-substituted. 1-6 Alkyl substitution;

[0027] n is an integer between 0 and 2.

[0028] In one embodiment of the invention, X is CH. In another embodiment of the invention, X is N.

[0029] In one embodiment of the invention, R 1 It is a halogen. In one type of embodiment, R 1 It is chlorine.

[0030] In one embodiment of the invention, R 2 It is a halogen. In one type of invention, R 2 It's fluorine.

[0031] In one embodiment of the invention, R 3 It is hydrogen. In another embodiment of the invention, R 3 It is a methyl group.

[0032] In one embodiment of the invention, R 4 It is hydrogen. In another embodiment of the invention, R 3 It is a methyl group.

[0033] In one embodiment of the invention, R 5 It is NH2. In another embodiment of the invention, R 5 It is OH.

[0034] In one embodiment of the invention, R 6 It is hydrogen.

[0035] In one embodiment of the invention, R 7 It is hydrogen.

[0036] In one embodiment of the invention, R 8 It is a phenyl group; wherein the phenyl group is optionally substituted with 1-3 substituents, the substituents being independently selected from halogens, R... x OR x NR 9 R 10 NR 9(C=O)R x NR 9 (C=O)OR x (C=O)NR 9 (C=O)OR x R y and OR y In one type of the described embodiments, R 8 It is a phenyl group; wherein the phenyl group is optionally substituted with 1-3 substituents, the substituents being independently selected from halogens, R... x OR x R y and OR y .

[0037] In one embodiment of the invention, n is 0. In another embodiment of the invention, n is 1. In yet another embodiment of the invention, n is 2.

[0038] The references to the above preferred classes and subclasses are intended to include all combinations of specific and preferred groups, unless otherwise stated.

[0039] Specific embodiments of the present invention include, but are not limited to, the compounds identified herein as Examples 1-133 or their pharmaceutically acceptable salts.

[0040] The scope of this invention also includes pharmaceutical compositions comprising a compound of Formula I as described above and a pharmaceutically acceptable carrier. The invention is also contemplated to cover pharmaceutical compositions comprising a pharmaceutically acceptable carrier and any compound specifically disclosed in this application. These and other aspects of the invention will be apparent from the teachings contained herein.

[0041] This invention includes compositions for treating diseases or conditions involving plasma kallikrein activity. Therefore, this invention includes compositions for treating visual impairment, diabetic retinopathy, diabetic macular edema, retinal vein occlusion, hereditary angioedema, diabetes, pancreatitis, cerebral hemorrhage, nephropathy, cardiomyopathy, neuropathy, inflammatory bowel disease, arthritis, inflammation, septic shock, hypotension, cancer, adult respiratory distress syndrome, disseminated intravascular coagulation, blood clotting during cardiopulmonary bypass surgery, and bleeding from postoperative surgery, comprising compounds of this invention in a pharmaceutically acceptable carrier. One class of this invention includes compositions for treating hereditary angioedema, uveitis, posterior uveitis, wet age-related macular edema, diabetic macular edema, diabetic retinopathy, and retinal vein occlusion. These compositions may optionally include anti-inflammatory agents, anti-VEGF agents, immunosuppressants, anticoagulants, antiplatelet agents, and thrombolytic agents. The composition can be added to blood, blood products, or mammalian organs to achieve the desired inhibition.

[0042] The present invention also includes compositions for the prevention or treatment of retinal vascular permeability associated with diabetic retinopathy and diabetic macular edema in mammals, comprising compounds of the present invention in a pharmaceutically acceptable carrier. These compositions may optionally include anti-inflammatory agents, anti-VEGF agents, immunosuppressants, anticoagulants, antiplatelet agents, and thrombolytic agents.

[0043] The present invention also includes compositions for treating inflammatory conditions of the eye, including, but not limited to, uveitis, posterior uveitis, macular edema, acute macular degeneration, wet age-related macular edema, retinal detachment, retinal vein occlusion, ocular tumors, fungal infections, viral infections, multifocal choroiditis, diabetic uveitis, diabetic macular edema, diabetic retinopathy, proliferative vitreoretinopathy, sympathetic ophthalmia, Vogt-Koyanagi-Harada syndrome, histoplasmosis, and uveitis. These compositions may optionally include anti-inflammatory agents, anti-VEGF agents, immunosuppressants, anticoagulants, antiplatelet agents, and thrombolytic agents.

[0044] The present invention also includes compositions for treating posterior eye diseases, including, but not limited to, uveitis, posterior uveitis, wet age-related macular edema, diabetic macular edema, diabetic retinopathy, and retinal vein occlusion. These compositions may optionally include anti-inflammatory agents, anti-VEGF agents, immunosuppressants, anticoagulants, antiplatelet agents, and thrombolytic agents.

[0045] It should be understood that the present invention relates to compounds of structural formula I described herein, pharmaceutically acceptable salts of compounds of structural formula I, and non-pharmaceutically acceptable salts when used as precursors of free compounds or their pharmaceutically acceptable salts, or in other synthetic operations.

[0046] The compounds of the present invention can be administered in the form of pharmaceutically acceptable salts. The term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic base or acid (including inorganic or organic bases and inorganic or organic acids). Salts of basic compounds included in the term "pharmaceutically acceptable salt" refer to non-toxic salts of the compounds of the present invention, which are typically prepared by reacting a free base with a suitable organic or inorganic acid. Representative salts of the basic compounds of the present invention include, but are not limited to, the following salts: acetate, ascorbate, adipic acid salt, alginate, aspirate, benzenesulfonate, benzoate, bicarbonate, bisulfate, tartrate, borate, bromide, butyrate, camphorate, camphor sulfonate, dextrorotatory camphor sulfonate, carbonate, chloride, clavulanate, citrate, cyclopentanepropionate, diethylacetate, digluconate, dihydrochloride, dodecylsulfonate, edetate, ethanedisulfonate, etopoate, esylate, ethanesulfonate, formate, fumarate, gluceptate, glucoheptanoate, gluconate, glutamate, glycerophosphate, glycolyl arsyl ... ilate), hemisulfate, heptanoate, hexanoate, hexylresorcinol salt, hydrabamine, hydrobromide, hydrochloride, 2-hydroxyethanesulfonate, hydroxynaphthylcarboxylate, iodide, isonicotinate, isothiosulfate, lactate, lactobionate, laurate, malate, maleate, mandelate, methanesulfonate, methyl bromide, methyl nitrate, methyl sulfate, methanesulfonate, mucilage, 2-naphthalenesulfonate, naphthalenesulfonate, nicotinate, nitrate, N 1. Methyl-reduced glucosamine ammonium salts, oleates, oxalates, pyrates (dihydroxynaphthyl salts), palmitates, pantothenates, pectates, persulfates, phosphates / bisphosphates, pimecrolates, phenylpropionates, polygalacturonic acids, propionates, salicylates, stearates, sulfates, basic acetates, succinates, tannates, tartrates, 8-chlorotheophylline salts, thiocyanates, toluenesulfonates, triethyliodide, trifluoroacetate, undeconate, valerates, etc. Furthermore, in cases where the compounds of the present invention carry an acidic moiety, suitable pharmaceutically acceptable salts include, but are not limited to, salts derived from inorganic bases, including aluminum, ammonium, calcium, copper, ferric iron, ferrous iron, lithium, magnesium, ferric manganese, ferrous manganese, potassium, sodium, zinc, etc.Salts derived from pharmaceutically acceptable organic non-toxic alkaloids include salts of the following substances: primary, secondary, and tertiary amines, cyclic amines, dicyclohexylamine, and basic ion exchange resins, such as arginine, betaine, caffeine, choline, N,N-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucosamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc. Furthermore, the included basic nitrogen-containing groups can be quaternized by reagents such as: lower alkyl halides, such as chlorides, bromides, and iodides of methyl, ethyl, propyl, and butyl; dialkyl sulfates such as dimethyl, diethyl, dibutyl, and dipentyl sulfates; long-chain halides, such as chlorides, bromides, and iodides of decyl, lauryl, myristyl, and stearyl; aralkyl halides such as bromides of benzyl and phenethyl, etc.

[0047] These salts can be obtained by known methods, such as by mixing the compounds of the present invention with an equal volume of a solution containing the desired acid, base, etc., and then collecting the desired salt by filtering the salt or distilling off the solvent. The compounds of the present invention and their salts can form solvates with solvents such as water, ethanol, or glycerol. Depending on the type of side-chain substituents, the compounds of the present invention can simultaneously form acid addition salts and base salts.

[0048] If the compound of Formula I contains both acidic and basic groups in its molecule, in addition to the salt forms mentioned, the present invention also includes internal salts or internal ammonium salts (betaines) (zwitterions).

[0049] This invention includes all stereoisomers of compounds of Formula I. Unless a specific stereochemistry is specified, this invention is intended to include all such isomers of these compounds. The asymmetric centers present in compounds of Formula I may each have an independent (R) configuration or (S) configuration. When the bond with the chiral carbon is described as a straight line in the structural formula of this invention, it should be understood that the (R) and (S) configurations of the chiral carbon and therefore each individual enantiomer and mixture thereof are included in the formula. When a particular configuration is described, it means that the enantiomer ((R) or (S), located at the center). Similarly, when the name of a compound is described without specifying the chirality of the chiral carbon, it should be understood that the (R) and (S) configurations of the chiral carbon and therefore each individual enantiomer and mixture thereof are included in the name. The generation of specific stereoisomers or mixtures thereof can be identified in instances where such stereoisomers or mixtures are obtained, but this in no way limits the inclusion of all stereoisomers and mixtures thereof within the scope of this invention.

[0050] Unless a specific enantiomer or diastereomer is specified, this invention encompasses all possible enantiomers and diastereomers, as well as mixtures of two or more stereoisomers in all ratios, such as mixtures of enantiomers and / or diastereomers. Thus, enantiomers in the following forms are the subject of this invention: enantiomerically pure forms, as levorotatory and dextrorotatory enantiomers, racemic forms, and mixtures of two enantiomers in all ratios. In the case of cis / trans isomerism, this invention includes cis and trans forms, as well as mixtures of these forms in all ratios. If desired, the preparation of individual stereoisomers can be carried out by separating the mixture using conventional methods (e.g., chromatography or crystallization), by using stereochemically consistent synthetic starting materials, or by stereoselective synthesis. Optionally, derivatization can be performed prior to the separation of stereoisomers. The separation of stereoisomer mixtures can be carried out as an intermediate step during the synthesis of compounds of Formula I, or on the final racemic product. Absolute stereochemistry can be determined by X-ray crystallography of the crystalline product or intermediate, which, if necessary, is derivatized with a reagent containing a stereocenter of known configuration. Where the compounds of the present invention are tautomerizable, all individual tautomers and mixtures thereof are included within the scope of the present invention. The present invention includes all such isomers, as well as salts, solvates (including hydrates), and solvated salts of such racemates, enantiomers, diastereomers, and tautomers, and mixtures thereof.

[0051] In the compounds of this invention, atoms may exhibit their natural isotopic abundance, or one or more atoms may be artificially enriched with specific isotopes having the same atomic number but different atomic masses or mass numbers from those that are predominantly found in nature. This invention is intended to include all suitable isotopic variants of the compounds specifically and generally described. For example, different isotopic forms of hydrogen (H) include protium (1... H ) and deuterium (2 H Protium is the dominant hydrogen isotope found in nature. Deuterium enrichment can provide certain therapeutic advantages, such as increasing in vivo half-life or reducing dosage requirements, or can provide compounds that can be used as standards for characterizing biological samples. Isotope-enriched compounds can be prepared without excessive experimentation using conventional techniques well known to those skilled in the art, or methods similar to those described in the general methodologies and examples herein, using appropriate isotope-enriching reagents and / or intermediates.

[0052] When any variable (e.g., R) xWhen a substituent (e.g., a variable) appears more than once in any component, its definition for each occurrence is independent of the others. Furthermore, combinations of substituents and variables are only permitted if such combinations produce stable compounds. Lines drawn from substituents into the ring system represent bonds that can be attached to any substituted ring atom. If the ring system is bicyclic, this means the bond is attached to any suitable atom on either ring of the bicyclic portion.

[0053] It should be understood that those skilled in the art can incorporate one or more silicon (Si) atoms into the compounds of the present invention to replace one or more carbon atoms, providing chemically stable compounds that can be readily synthesized from readily available starting materials using techniques known in the art. When comparing similar C and Si element bonds, carbon and silicon have different covalent radii, leading to differences in bond lengths and spatial arrangement. These differences result in subtle variations in the size and shape of silicon-containing compounds compared to carbon. Those skilled in the art will understand that differences in size and shape can lead to subtle or significant variations in potency, solubility, off-target activity, packaging properties, etc. (Diass, JO et al., Organometallics (2006) 5: 1188-1198; Showell, GA et al., Bioorganic & Medicinal Chemistry Letters (2006) 16: 2555-2558).

[0054] It should be understood that those skilled in the art can choose the substituents and substitution patterns of the compounds of the present invention to provide chemically stable compounds that can be readily synthesized from readily available starting materials using techniques known in the art and the methods set forth below. If the substituent itself is substituted by more than one group, it should be understood that these multiple groups may be on the same carbon or on different carbons, as long as a stable structure is produced. The phrase "optionally substituted (by one or more substituents)" should be understood to mean that the group in question is unsubstituted or may be substituted by one or more substituents.

[0055] Furthermore, the compounds of the present invention can exist in amorphous and / or one or more crystalline forms, and therefore all amorphous and crystalline forms of the compounds of Formula I and mixtures thereof are intended to be included within the scope of the present invention. Additionally, some compounds of the present invention can form solvates with water (i.e., hydrates) or common organic solvents. Such solvates and hydrates of the compounds of the present invention (particularly pharmaceutically acceptable solvates and hydrates), as well as non-solventized and anhydrous forms, are also included within the scope of the present invention.

[0056] Furthermore, in the presence of a carboxylic acid (-COOH) or alcohol group in the compounds of the present invention, pharmaceutically acceptable esters of carboxylic acid derivatives, such as methyl, ethyl, or neopentanoyloxymethyl, or acyl derivatives of alcohols, such as O-acetyl, O-neopentyl, O-benzoyl, and O-aminoacyl, may be used. This includes those esters and acyl groups known in the art for altering the solubility or hydrolytic characteristics of formulations used for sustained release or prodrug preparations.

[0057] Any pharmaceutically acceptable prodrug modification of the compounds of the present invention (which results in in vivo conversion to compounds within the scope of the present invention) is also within the scope of the present invention. For example, esters can optionally be prepared by esterification of a available carboxylic acid group or by forming an ester on a available hydroxyl group in the compound. Similarly, unstable amides can be prepared. Pharmaceutically acceptable esters or amides of the compounds of the present invention can be prepared as prodrugs, which can be hydrolyzed back to acid (or -COO). - (depending on the pH of the liquid or tissue in which the transformation occurs) or hydroxyl form, particularly in vivo, and therefore included within the scope of this invention. Examples of pharmaceutically acceptable prodrug modifications include, but are not limited to, -C 1-6 Alkyl esters and -C substituted with phenyl esters 1-6 alkyl.

[0058] Therefore, the compounds in the general structural formulas, embodiments and specific compounds described and claimed herein include their salts, all possible stereoisomers and tautomers, physical forms (e.g., amorphous and crystalline forms), solvates and hydrates, and any combination of these forms, as well as their salts, their prodrug forms and salts of their prodrug forms, provided such forms are possible, unless otherwise stated.

[0059] Unless otherwise stated herein, the terms “alkyl” and “alkylene” are intended to include branched and straight-chain saturated aliphatic hydrocarbon groups having a specified number of carbon atoms. Common abbreviations for alkyl, such as methyl, are used throughout the specification and may be represented by symbols including “Me” or CH3 or as extended bonds (e.g., “…”). The common abbreviations, including ""), are used to represent ethyl, which can be represented by "Et" or CH2CH3; propyl can be represented by "Pr" or CH2CH2CH3; butyl can be represented by "Bu" or CH2CH2CH2CH3, and so on. 1-4 Alkyl (or "C1-C4 alkyl") refers, for example, a straight-chain or branched alkyl group having a specified number of carbon atoms, including all isomers. For example, structure...

[0060]

[0061] They have the same meaning. C 1-4Alkyl groups include n-butyl, isobutyl, sec-butyl, and tert-butyl, n-propyl and isopropyl, ethyl, and methyl. If no number is specified, straight-chain or branched alkyl groups consist of 1 to 4 carbon atoms.

[0062] Unless otherwise specified, the term "cycloalkyl" refers to a monocyclic or bicyclic saturated aliphatic hydrocarbon group having a specified number of carbon atoms. For example, "cycloalkyl" includes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0063] Unless otherwise specified, the term "aryl" as used herein refers to a stable monocyclic or bicyclic ring system having up to 10 carbon atoms in each ring, wherein at least one ring is aromatic. Bicyclic aryl ring systems include fused ring systems in which two rings share two atoms; and spirocyclic systems in which two rings share one atom. Aryl groups within the scope of this definition include, but are not limited to, phenyl, indene, isoindene, naphthalene, and tetrahydronaphthalene.

[0064] Unless otherwise specified, the term "heteroaryl" as used herein refers to a stable monocyclic or bicyclic ring system having up to 10 atoms in each ring, wherein at least one ring is aromatic and at least one ring contains 1 to 4 heteroatoms selected from O, N, and S. Bicyclic heteroaryl ring systems include fused ring systems in which two rings share two atoms; and spirocyclic systems in which two rings share one atom. The heteroaryl groups within the scope of this definition include, but are not limited to: azaindolyl, benzimidazolyl, benzisoxazolyl, benzofuranyl, benzofurazanyl, benzopyrazolyl, benzotriazolyl, benzothiophenyl, benzoxazolyl, carbazoyl, carbaolinyl, cenolinyl, dihydroindenyl, furanyl, indololinyl, indolyl, indazinyl, indazolyl, indazolyl, isobenzofuranyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, naphthyl, naphthidyl, oxadiazolyl, oxazolyl, oxazoline, isoxazoline, pyranyl, pyrazinyl, pyrazolyl, pyrazolopyrimidinyl, pyridazinyl, pyridinyl, pyrimidinyl, pyrroleyl, quinazolinyl, quinolinyl Quinoxolinyl, tetrazolyl, tetrazolopyridyl, thiadiazolyl, thiazolyl, thiophene, triazolyl, dihydrobenzimidazolyl, dihydrobenzofuranyl, dihydrobenzothiophene, dihydrobenzooxazolyl, dihydroindolyl, dihydroquinolinyl, dihydrobenzodioxazinyl, dihydropyrazolyothiazinedioxidyl, methylenedioxybenzene, benzothiazolyl, benzothiaphene, quinolinyl, isoquinolinyl, oxazolyl, tetrahydroquinolinyl, sulfolane, 1,3-benzodioxazopentenyl, and 3-oxo-3,4-dihydro-2N-benzo[b][1,4]thiazine. If a heteroaryl group contains a nitrogen atom, it should be understood that its corresponding N-oxide is also included in this definition.

[0065] As used herein, the term "heterocyclic" or "heterocyclic group" means a stable non-aromatic monocyclic or bicyclic ring system having at most 10 atoms in each ring, and, unless otherwise indicated, containing 1-4 heteroatoms selected from O, N, S, SO, or SO2. Bicyclic heterocyclic systems include fused ring systems, where two rings share two atoms; and spirocyclic systems, where two rings share one atom. Therefore, "heterocyclic group" includes, but is not limited to, the following: azaspirononyl, azaspironoctyl, azacyclobutyl, dioxacyclohexyl, oxadiaspirondecenyl, oxaspironoctyl, oxazolidinone, piperazine, piperidinyl, pyrrolyl, morpholinyl, thiomorpholinyl, tetrahydrofuranyl, tetrahydropyranyl, dihydropiperidinyl, tetrahydrothiophene, etc. If the heterocycle contains nitrogen, it should be understood that its corresponding N-oxide is also included in this definition.

[0066] Unless otherwise specified, the terms “halogen” or “halogenated” refer to fluorine, chlorine, bromine, or iodine.

[0067] (Fluka) diatomaceous earth is diatomaceous earth and can also be called "celite".

[0068] Unless otherwise stated herein, structures containing substituent variables such as the variable "R" below (which is described as not being attached to any particular bicyclic carbon atom) are:

[0069]

[0070] This represents a structure in which the variable can optionally be attached to any bicyclic carbon atom. For example, the variable R shown in the structure above can be attached to any one of the six bicyclic carbon atoms i, ii, iii, iv, v, or vi.

[0071] Unless otherwise stated herein, bicyclic ring systems include fused ring systems in which two rings share two atoms; and spirocyclic systems in which two rings share one atom.

[0072] The present invention also relates to a medicament comprising at least one compound of formula I and / or a pharmaceutically acceptable salt of a compound of formula I and / or optionally a stereoisomer of a compound of formula I or a pharmaceutically acceptable salt of a stereoisomer of a compound of formula I, as well as a pharmaceutically suitable and pharmaceutically acceptable carrier, additive and / or other active substance and adjuvant.

[0073] The term "patient" as used in this article refers to mammals such as primates, humans, sheep, horses, cattle, pigs, dogs, cats, rats, and mice.

[0074] The medicament according to the invention can be administered orally, by inhalation, rectal or transdermal application, or by subcutaneous, intra-articular, intraperitoneal or intravenous injection. Oral administration is preferred. The scaffold and other surfaces in contact with blood in the body can be coated with compounds of Formula I.

[0075] The present invention also relates to a method for producing a pharmaceutical product, comprising preparing at least one compound of formula I into a suitable administration form using a pharmaceutically suitable and pharmaceutically acceptable carrier and optionally other suitable active substances, additives or adjuvants.

[0076] Suitable solid or galenal formulations include, for example, granules, powders, coated tablets, tablets, (micro)capsules, suppositories, syrups, juices, suspensions, emulsions, drops, or injectable solutions, as well as formulations with prolonged release of the active substance, prepared using conventional excipients such as mediators, disintegrants, binders, coating agents, swelling agents, flow aids or lubricants, flavoring agents, sweeteners, and solubilizers. Commonly used excipients that may be mentioned include magnesium carbonate, titanium dioxide, lactose, mannitol and other sugars, talc, gelatin, starch, cellulose and its derivatives, animal and vegetable oils such as cod liver oil, sunflower oil, peanut oil or sesame oil, polyethylene glycol, and solvents such as, for example, sterile water, and monohydric or polyhydric alcohols such as glycerin.

[0077] The dosage regimen for plasma kallikrein inhibitors is selected based on several factors, including patient type, race, age, weight, sex, and medical condition; the severity of the condition to be treated; the route of administration; the patient's renal and hepatic function; and the specific compound or its salt used. A general practitioner or veterinarian can easily determine and prescribe the effective amount of medication needed to prevent, counteract, or halt disease progression.

[0078] When used for a specified effect, the oral dose range of a plasma kallikrein inhibitor is from about 0.01 mg / kg body weight / day (mg / kg / day) to about 30 mg / kg / day, preferably 0.025-7.5 mg / kg / day, more preferably 0.1-2.5 mg / kg / day, and most preferably 0.1-0.5 mg / kg / day (unless otherwise stated, the amount of active ingredient is based on free base). For example, an 80 kg patient would receive about 0.8 mg / day to 2.4 g / day, preferably 2-600 mg / day, more preferably 8-200 mg / day, and most preferably 8-40 mg / kg / day. Therefore, a suitably prepared medicine for once-daily administration contains 0.8 mg to 2.4 g, preferably 2 mg to 600 mg, more preferably 8 mg to 200 mg, and most preferably 8 mg to 40 mg, for example, 8 mg, 10 mg, 20 mg, and 40 mg. Advantageously, plasma kallikrein inhibitors can be administered in fractionated doses twice, three, or four times daily. For twice-daily administration, suitably prepared drugs contain 0.4 mg to 4 g, preferably 1 mg to 300 mg, more preferably 4 mg to 100 mg, and most preferably 4 mg to 20 mg, for example, 4 mg, 5 mg, 10 mg, and 20 mg.

[0079] Intravenous administration delivers an amount sufficient to deliver 0.025-7.5 mg / kg / day, preferably 0.1-2.5 mg / kg / day, and more preferably 0.1-0.5 mg / kg / day of the active ingredient. Such amounts can be administered in a variety of suitable manners, such as large volumes of low-concentration active ingredient administered over a prolonged period or several times daily, or small volumes of high-concentration active ingredient administered over a short period, such as once daily. Typically, conventional intravenous formulations can be prepared containing an active ingredient at a concentration of about 0.01-1.0 mg / mL, such as 0.1 mg / mL, 0.3 mg / mL, and 0.6 mg / mL, and administered daily in amounts ranging from 0.01 mL / kg of patient weight to 10.0 mL / kg of patient weight, such as 0.1 mL / kg, 0.2 mL / kg, and 0.5 mL / kg. In one embodiment, an 80 kg patient receiving 8 mL of an intravenous formulation containing an active ingredient at a concentration of 0.5 mg / mL twice daily receives 8 mg of the active ingredient daily. Glucuronic acid, L-lactic acid, acetic acid, citric acid, or any pharmaceutically acceptable acid / conjugate base with reasonable buffering capacity within an acceptable pH range for intravenous administration can be used as a buffer. Depending on the solubility of the drug to be administered, a person skilled in the art can readily make an appropriate choice of buffer and pH for the formulation.

[0080] The compounds of Formula I can be administered as a monotherapy or in combination with other therapeutic agents, including, but not limited to, anti-inflammatory agents, anti-VEGF agents, immunosuppressants, anticoagulants, antiplatelet agents, and thrombolytic agents.

[0081] "Anti-inflammatory agents" are any medications that, when administered at therapeutically effective levels, directly or indirectly reduce inflammation. "Anti-inflammatory agents" include, but are not limited to, steroidal anti-inflammatory agents and glucocorticoids. Suitable anti-inflammatory agents include, but are not limited to, cortisone, dexamethasone, hydrocortisone, methylprednisolone, prednisolone, prednisolone, and triamcinolone.

[0082] "Anti-VEGF agents" are any drugs that directly or indirectly inhibit the activity of VEGF (vascular endothelial growth factor). Suitable anti-VEGF agents include, but are not limited to, bevacizumab, ranibizumab, and aflibercept.

[0083] "Immunosuppressants" are any medications that directly or indirectly suppress or reduce the strength of the body's immune system. Suitable immunosuppressants include, but are not limited to, corticosteroids (e.g., prednisone, budesonide, prednisolone), janus kinase inhibitors (e.g., tofacitinib), calcineurin inhibitors (e.g., cyclosporine, tacrolimus), mTOR inhibitors (e.g., sirolimus, everolimus), IMDH inhibitors (e.g., azathioprine, leflunomide, mycophenolate mofetil), biologics (e.g., abatacept, adalimumab, anaprole, cetuzumab, etanercept, golimumab, infliximab, ixekizumab, natezilumab, rituximab, secukinumab, tocilizumab, ustekinumab, vedolizumab), and monoclonal antibodies (e.g., baliximab, dacrolimus).

[0084] Suitable anticoagulants include, but are not limited to, factor XIa inhibitors, thrombin inhibitors, thrombin receptor antagonists, factor VIIa inhibitors, factor Xa inhibitors, factor IXa inhibitors, factor XIIa inhibitors, adenosine diphosphate antiplatelet agents (e.g., P2Y12 antagonists), fibrinogen receptor antagonists (e.g., to treat or prevent unstable angina or to prevent restenosis and re-occlusion after angioplasty), other anticoagulants such as aspirin, and thrombolytic agents such as plasminogen activators or streptokinase, to achieve synergistic effects in the treatment of various vascular diseases. Such anticoagulants include, for example, apixaban, dabigatran, cangreloxol, ticagrelor, vorapazol, clopidogrel, edoxaban, mipomisen, prasugrel, rivaroxaban, and sermoheparin. For example, patients with coronary artery disease and those undergoing angioplasty will benefit from the co-administration of fibrinogen receptor antagonists and thrombin inhibitors.

[0085] In some embodiments, the anti-inflammatory agents, anti-VEGF agents, immunosuppressants, anticoagulants, antiplatelet agents, and thrombolytics described herein are used in their conventional dosage ranges and regimens as reported in the art, including, for example, the dosages described in the following literature: Physicians' Desk Reference Various versions, such as version 70 (2016) and earlier. In other embodiments, the anti-inflammatory agents, anti-VEGF agents, immunosuppressants, anticoagulants, antiplatelet agents, and thrombolytic agents described herein are used at doses below their usual dose range.

[0086] Alternatively or additionally, one or more additional pharmacologically active agents may be administered in combination with the compounds of the present invention. The additional active agents (or agents in combination) are defined as pharmaceutically active agents (or agents in combination) that are active in vivo, including prodrugs that are converted to a pharmaceutically active form after administration, which differ from the compounds of the present invention, and, when such a form is commercially available or otherwise chemically possible, also include free acids, free bases, and pharmaceutically acceptable salts of the additional active agents. Generally, any suitable one or more additional active agents (including, but not limited to, antihypertensive agents, additional diuretics, antiatherosclerotic agents such as lipid-regulating compounds, antidiabetic agents, and / or antiobesity agents) may be used in any combination with the compounds of the present invention in a single-dose formulation (fixed-dose drug combination), or may be administered to the patient in one or more separate-dose formulations, which allows for simultaneous or sequential administration of the active agents (co-administration of individual active agents). Examples of other active agents that may be used include, but are not limited to, angiotensin-converting enzyme inhibitors (e.g., alapril, benazepril, captopril, siropril, cilazapril, delapril, enalapril, enalapril, fosinopril, imidapril, lisinopril, muvipril, perindopril, quinapril, ramipril, spiropril, temopril, or quindopril); angiotensin II receptor antagonists, also known as angiotensin receptor blockers or ARBs, which may be in the form of a free base, free acid, salt, or prodrug, such as azilsartan, for example, meazartan potassium. Candesartan, for example, candesartan medoxomil eprosartan, for example, eprosartan mesylate Irbesartan Losartan, for example, losartan potassium Olmesartan, for example, olmesartan medoxomil Telmisartan Valsartan and with thiazide-like diuretics such as hydrochlorothiazide (e.g., DIOVAN ATACAND Any of these drugs used in combination (e.g., milloride, spironolactone, eplerenone, triamterene, each with or without HCTZ); neutral endopeptidase inhibitors (e.g., thiorphan and phosphonyl dipeptide); aldosterone antagonists; aldosterone synthase inhibitors; renin inhibitors; enalkrein; RO 42-5892; A 65317; CP 80794; ES 1005; ES 8891; SQ 34017; Aliskiren (2(S),4(S),5(S),7(S)-N-(2-carbamoyl-2-methylpropyl)-5-amino-4-hydroxy-2,7-diisopropyl-8-[4-methoxy-3-(3-methoxypropoxy)-phenyl]octamido hemifumarate) SPP600, SPP630 and SPP635); endothelin receptor antagonists; vasodilators (e.g. nitroprusside); calcium channel blockers (e.g. amlodipine, nifedipine, verapamil, diltiazem, felodipine, golopamil, niludipine, nimodipine, nicardipine); potassium channel activators (e.g. Examples include nicorandil, pinacidil, simvadin, minoxidil, alpracaline, and cloprazolam; sympathetic nerve blockers; β-adrenergic blockers (e.g., acebutolol, atenolol, betalol, bisoprolol, carvedilol, metoprolol, metoprolol tartrate, nadolol, propranolol, sotalol, and timolol); α-adrenergic blockers (e.g., doxazosin, prazosin, or α-methyldopa); central α-adrenergic agonists; peripheral vasodilators (e.g., hydralazine); and lipid-lowering agents, such as HMG-CoA reductase inhibitors like simvastatin and lovastatin (which are available as lactone prodrugs). Know (Sold and acting as an inhibitor after administration), and pharmaceutically acceptable salts of dihydroxyocyclocarboxylic acid HMG-CoA reductase inhibitors such as atorvastatin (especially in the form of...). calcium salts for sale), rosuvastatin (especially in the form of calcium salts), calcium salts sold), pravastatin (especially with Sodium salts sold) and fluvastatin (especially in the form of sodium salts) and fluvastatin (especially in the form Sodium salts sold in stores; cholesterol absorption inhibitors such as ezetimibe. And with any other lipid-lowering agents such as HMG-CoA reductase inhibitors mentioned above, and especially with simvastatin Ezetimibe in combination with atorvastatin calcium; nicotinic acid in immediate-release or controlled-release forms, and especially in combination with DP antagonists such as laropieren and / or with HMG-CoA reductase inhibitors; nicotinic acid receptor agonists such as acilimus and acyclofuran, and partial nicotinic acid receptor agonists; metabolic modifiers, including insulin sensitizers and related compounds used to treat diabetes such as biguanides (e.g., metformin), chloropicrins (e.g., repaglinide, nateglinide), sulfonylureas (e.g., chlorpropamide, glimepiride, glipizide, glibenclamide, tolazoline, tolbutamide), thiazolidinediones also known as glitazones (e.g., pioglitazone, rosiglitazone), alpha-glucosidase inhibitors (e.g., acarbose, miglitol), dipeptidyl peptidase inhibitors (e.g., sitagliptin) Alogliptin, vildagliptin, saxagliptin, liraliptin, dugliptin, giglitazone), ergot alkaloids (e.g., bromocriptine), and combination drugs such as (Sitagliptin and metformin) and injectable diabetes medications such as exenatide and prulaminide acetate; glucose uptake inhibitors, such as sodium-glucose transporter (SGLT) inhibitors and their various isoforms, such as SGLT-1, SGLT-2 (e.g., ASP-1941, TS-071, BI-10773, tolpagliflozin, LX-4211, canagliflozin, dapagliflozin, elepagliflozin, etagliflozin, repaggliflozin, and soragliflozin) and SGLT-3; or other medications beneficial for the prevention or treatment of the aforementioned diseases, including, but not limited to, diazoxide; and including free acids, free bases, and pharmaceutically acceptable salt forms, prodrug forms, such as, where chemically possible, esters and salts of the prodrugs of the above-mentioned medications. Trademark names of the above-mentioned medications are provided to illustrate the sales forms of the active agents; such medications may be used in single dosage forms to be administered simultaneously or sequentially with the compounds of the present invention, or one or more of the active agents may be used in a fixed-dose combination of medications comprising the compounds of the present invention.

[0087] Typical doses of the plasma kallikrein inhibitors of the present invention, when combined with other suitable agents, may be the same as, or significantly lower than, those doses of plasma kallikrein inhibitors administered without co-administration of other agents, depending on the patient’s treatment needs.

[0088] The compound is administered to mammals in a therapeutically effective amount. A "therapeutically effective amount" means an amount of the compound of the invention that, when administered to mammals alone or in combination with other therapeutic agents, is effective in treating (i.e., preventing, inhibiting, or improving) a disease condition or in treating the progression of disease in the host.

[0089] The compounds of the present invention are preferably administered to mammals alone in a therapeutically effective amount. However, the compounds of the present invention may also be administered to mammals in combination with other therapeutic agents in a therapeutically effective amount, as described below. When administered in combination, the combination of compounds is preferably, but not necessarily, a synergistic combination. A synergistic effect occurs when the effect of the compounds (in this application, inhibition of the desired target) when administered in combination is greater than the additive effect of each compound when administered as a single agent, as described, for example, as Chou and Talalay, Adv. Enzyme Regul. 1984, 22, 27-55. Generally, synergistic effects are most clearly demonstrated at suboptimal concentrations of the compounds. Compared to individual components, synergistic effects can be reduced cytotoxicity, increased anticoagulant activity, or some other beneficial effect of the combination.

[0090] "Combination administration" or "combination therapy" refers to the parallel administration of the compounds of the present invention and one or more other therapeutic agents to a mammal being treated. When administered in combination, each component may be administered simultaneously or sequentially at different times in any order. Thus, each component may be administered individually, but close enough in time to provide the desired therapeutic effect. The administration of each component does not need to be via the same route of administration; for example, one component may be administered orally, and another component may be delivered into the vitreous humor of the eye.

[0091] This invention is not limited in scope to the specific embodiments disclosed in the examples, which are intended to illustrate several aspects of the invention, and any functionally equivalent embodiments are within the scope of this invention. In fact, various modifications to the invention, in addition to those shown and described herein, will become apparent to those skilled in the art and are intended to fall within the scope of the appended claims.

[0092] General methods

[0093] The compounds of the present invention can be prepared using conventional techniques or according to the methods outlined in the general synthetic schemes below. Those skilled in the art can modify the shown procedures and reagents to obtain similar intermediates and / or final compounds.

[0094] NMR spectra were measured on VARIAN or Bruker NMR Systems (400, 500, or 600 MHz). Chemical shifts were reported in ppm low-field and high-field values ​​from tetramethylsilane (TMS), with reference to internal TMS or solvent resonances. 1 H NMR: δ7.27 (for CDCl3), δ2.50 (for (CD3)(CHD2)SO, and 13C18 NMR: δ77.02 (for CDCl3), δ39.51 (for (CD3)2SO). Coupling constants (J) are expressed in Hertz (Hz), and spin multiplicity is expressed as s (singleton), d (doublet), dd (doublet), t (triplet), m (multiplet), and br (broad peak). Chiral resolution was performed on a Waters Thar 80 SFC or Berger MG II preparative SFC system. LC-MS data were recorded using a C18 column with a MeCN gradient in water containing 0.02 to 0.1% TFA on a SHIMADAZU LC-MS-2020, SHIMADAZU LC-MS-2010, or Agilent 1100 series LC-MS, Agilent Prime-1260, or Waters Acquity LC-MS instrument. UV detection was performed at 220 and / or 254 nm, and ESI ionization was used for MS detection.

[0095] When chiral resolution is achieved by chromatography using chiral columns, the chiral columns used for SFC chiral resolution are listed in the table. Some of the chiral columns used are CHIRALPAK AD, CHIRALCEL OJ, CHIRALPAK AS, CHIRALPAK AY, CHIRALPAK IA, CHIRALPAK AD-H, and CHIRALPAK AS-H. These chiral columns will then be indicated by two or three-letter abbreviations. By convention, the fast-eluting isomer from the chiral resolution is always listed first in the table, followed by the slower-eluting isomer from the same resolution. If more than two isomers are separated, they will always be listed in the table in their elution order, such as peak 1, then peak 2, peak 3, and so on. An asterisk (*) near the chiral center in the structure indicates that the chiral center was resolved by chiral resolution, and its stereochemical configuration is not definitively determined.

[0096] Furthermore, TLC is thin-layer chromatography; UV is ultraviolet; W is watts; wt% is weight percentage; xg is a multiple of gravity; α D Specific rotation of polarized light at 589 nm; ℃ is degrees Celsius; %w / v is the percentage of the weight of the former reagent relative to the volume of the latter reagent; Hz is Hertz; cpm is counts per minute; δ H d is chemical shift; dd is doublet; MHz is megahertz; MS is mass spectrometry, and the mass spectrometry obtained by ES-MS can be referred to as "LC-MS" in this paper; m / z is mass-to-charge ratio; n is positive; N is equivalent concentration; nm is nanometer; nM is nanomolar concentration.

[0097] For the purposes of this specification, the following abbreviations have the indicated meanings: [Mes-Acr-Me] + is 9-mesityl-10-methylacridinium tetrafluoroborate; X-PHOS Pd G2 is chloro(2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II).

[0098] Ac acetyl

[0099] ACN acetonitrile

[0100] AcOH acetic acid

[0101] aq. aqueous

[0102] Boc or BOC tert-butoxycarbonyl

[0103] br broad peak

[0104] Bu or n Bu (n-)butyl

[0105] Bz benzoyl

[0106] ℃ degree Celsius

[0107] calcd. calculated

[0108] δ chemical shift

[0109] d doublet

[0110] DAST (diethylamino)sulfur trifluoride

[0111] DBU 1,8-diazabicyclo[5.4.0]undec-7-ene

[0112] DCM dichloromethane

[0113] dd doublet of doublets

[0114] DIEA, DIPEA N,N-diisopropylethylamine or Hünig's base

[0115] DMF dimethylformamide

[0116] DMSO dimethyl sulfoxide

[0117] DPPA diphenylphosphoryl azide

[0118] dqd doublet of double quartets

[0119] DTT dithiothreitol

[0120] EDC 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide

[0121] EDTA (ethylenediaminetetraacetic acid)

[0122] Equivalent

[0123] ESI Electrospray Ionization

[0124] Et Ethyl

[0125] Et2O diethyl ether

[0126] EtOH (ethanol)

[0127] EtOAc (ethyl acetate)

[0128] g gram

[0129] GST glutathione S-transferase

[0130] h hours

[0131] HATU N,N,N′,N′-Tetramethyl-O-(7-azabenzotriazol-1-yl)

[0132] Ureonium hexafluorophosphate

[0133] HPLC (High Performance Liquid Chromatography)

[0134] Hz Hertz

[0135] IPA isopropanol

[0136] i Pr isopropyl

[0137] J Coupling constant

[0138] LC liquid chromatography

[0139] LCMS (Liquid Chromatography-Mass Spectrometry)

[0140] LED Light Emitting Diode

[0141] m multiplet

[0142] M molar concentration

[0143] Me methyl

[0144] MeOH (methanol)

[0145] mg

[0146] MHz

[0147] min minutes

[0148] μL

[0149] mL

[0150] mM millimolecular concentration

[0151] mmol millimole

[0152] MS mass spectrometry

[0153] MTBE (methyl tert-butyl ether)

[0154] N-substituted

[0155] nm nanometer

[0156] nM nanomolar concentration

[0157] NMP 1-methylpyrrolidone

[0158] NMR (Nuclear Magnetic Resonance) spectroscopy

[0159] OAc acetate

[0160] Ph phenyl

[0161] Pr propyl

[0162] q Quadruple Peak

[0163] RAC racemic mixture

[0164] RT or rt refers to room temperature (ambient temperature, approximately 25°C).

[0165] s single peak

[0166] satd. saturated

[0167] SFC Supercritical Fluid Chromatography

[0168] t triple peak

[0169] T3P propanephosphonic anhydride

[0170] TBAF tert-butylammonium fluoride

[0171] TBS or TBDMS tert-butyldimethylsilyl

[0172] TBSCl tert-butyldimethylchlorosilane

[0173] t Bu tert-butyl

[0174] t BuOH tert-butanol

[0175] TCFH Tetramethylchloromethylammonium hexafluorophosphate

[0176] TEA (Triethylamine, Et3N)

[0177] Uncle Tert

[0178] TFA (trifluoroacetic acid)

[0179] THF Tetrahydrofuran

[0180] TLC (Thin Layer Chromatography)

[0181] TMS (trimethylsilyl)

[0182] TMSCl Trimethylchlorosilane

[0183] Tris(hydroxymethyl)aminomethane

[0184] Ts Toluenesulfonyl (Toluyl)

[0185] tt Triple Peak

[0186] X-phos or X-PHOS 2-Dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl

[0187] General methods

[0188] The starting materials used are derived from commercial sources or prepared in other embodiments, unless otherwise indicated. The methods for preparing the compounds of the present invention are exemplified by the following schemes. Unless otherwise stated, all starting materials used are commercially available.

[0189] Chiral separation method

[0190] The general preparation conditions for separating diastereomers or diastereomer mixtures of compounds using chiral SFC are as follows:

[0191]

[0192]

[0193] General Plan

[0194] Option A.

[0195]

[0196] Scheme A illustrates a synthetic sequence for preparing substituted spirocarbamates such as A6 from Boc-protected aniline A1 and ketones such as A2. Directed lithiation of aniline A1 and addition to the heterocyclic ketone A2 occur in the presence of a Lewis acid (e.g., LaCl3). The tertiary alcohol undergoes in-situ cyclization on the carbamate to generate a spirocarbamate derivative such as A3, which can be chirally separated, preferably using supercritical fluid chromatography (SFC) to provide enantiomers A4 and A5. Deprotection of either enantiomer (e.g., A4) yields the secondary amine A6.

[0197] Option B.

[0198]

[0199] Scheme B illustrates the synthetic sequence for preparing alkyl hydrazides such as B4 from carbonyl derivatives such as B1. The condensation of carbonyl B1 with benzoyl hydrazide yields intermediate B2, which is reduced to the protected hydrazine B3. Deprotection under acidic conditions yields alkyl hydrazine B4.

[0200] Option C.

[0201]

[0202] Scheme C illustrates the synthetic sequence for preparing CF3-ethyl hydrazine derivatives such as C4 from carbonyl derivatives such as C1. The condensation of carbonyl C1 with benzoyl hydrazine yields intermediate C2. CF3 is added to C2 with TMSCF3, providing CF3-ethyl intermediate C3, which is then deprotected under acidic conditions to yield the hydrazine derivative C4.

[0203] Option D.

[0204]

[0205] Scheme D illustrates the synthesis of alkyl hydrazine derivatives such as D2 from alkyl halides such as D1. Alkylation of hydrazine with alkyl halide D1 yields alkyl hydrazine D2.

[0206] Option E.

[0207]

[0208] Scheme E illustrates the synthetic sequence for preparing alkyl hydrazine derivatives such as E3 from carboxylic acid derivatives such as E1. Photoredox decarboxylhydrazidelation of carboxylic acid E1 provides a protected hydrazine intermediate E2. Deprotection yields the alkyl hydrazine derivative E3.

[0209] Option F.

[0210]

[0211] Scheme F illustrates a synthetic sequence for preparing alkyl hydrazine derivatives such as F4 from alkyl carboxylic acid derivatives such as F1. The Curtius rearrangement of alkyl carboxylic acid F1 yields a protected amine F2. Oxidation of F2 provides an N-nitroso intermediate F3, which is reduced and deprotected to produce the alkyl hydrazine derivative F4.

[0212] Option G.

[0213]

[0214] Scheme G illustrates the synthesis of arylhydrazines such as G2 from aryl bromides such as G1. Arylhydrazine G2 is prepared by a palladium-catalyzed cross-coupling reaction of aryl bromides such as G1 with hydrazine in the presence of a suitable base.

[0215] Option H.

[0216]

[0217] Scheme H illustrates the synthetic sequence for preparing aminopyrazole derivatives such as H4 from hydrazine derivatives such as H2 and cyanoalkoxyacrylates H1. The condensation of the substituted hydrazine H2 with cyanoethoxyacrylate H1 provides an ester-aminopyrazole derivative such as H3. The saponification of H3 yields a carboxylic acid H4.

[0218] Option I.

[0219]

[0220] Scheme I illustrates the synthetic sequence for preparing hydroxypyrazole derivatives such as I4 from hydrazine derivatives such as I2 and alkoxymethylene malonate I1. The condensation of substituted hydrazine I2 with malonate I1 provides an ester hydroxypyrazole derivative such as I3. Saponification of I3 yields carboxylic acid I4.

[0221] Option J.

[0222]

[0223] Scheme J illustrates the synthetic sequence for preparing N-substituted aminopyrazole derivatives, such as J4, from unsubstituted aminopyrazole derivatives, such as J1. Alkylation of aminopyrazole J1 yields N-substituted aminopyrazole J3. Saponification of J3 yields the carboxylic acid J4 of the N-substituted aminopyrazole.

[0224] Option K.

[0225]

[0226] Scheme K illustrates the preparation of spirocarbamate pyrazole or triazole derivatives such as K1 from spirocarbamate piperidine derivatives such as K1 and carboxylic acid derivatives such as K2. The carboxylic acid K2 is coupled to the spirocarbamate K1 using a peptide coupling agent such as TCFH, EDC, HATU, or T3P to provide amide K3.

[0227] Option L.

[0228]

[0229] Scheme L illustrates a synthetic sequence for preparing spirocarbamate aminopyrazole derivatives such as L5 from spirocarbamate piperidine derivatives such as L1. The coupling of spirocarbamate L1 with cyanoacetic acid provides intermediate L2, which undergoes condensation to generate cyanoacrylate derivatives such as L3. Condensation of L3 with various hydrazides L4 under basic or acidic conditions provides aminopyrazoles such as L5.

[0230] Intermediate A2-1

[0231]

[0232] 3-Cyclopropyl-5-oxoperidin-1-carboxylic acid tert-butyl ester

[0233] Cyclopropylmagnesium bromide (30.0 mL, 15.2 mmol, 0.5 M in THF) was added to a suspension of CuI (1.45 g, 7.61 mmol) in THF (20 mL) at -78 °C under a nitrogen atmosphere. The resulting mixture was warmed to 0 °C and stirred for another 1 h to produce an organocubic reagent. The solution was then cooled to -78 °C, followed by the addition of a THF solution (5 mL) of tert-butyl 3-oxo-3,6-dihydropyridine-1(2H)-carboxylate (1.0 g, 5.1 mmol) and TMSCl (1.30 mL, 10.1 mmol). The reaction mixture was stirred at -78 °C for 1 h and then quenched with MeOH. The mixture was diluted with EtOAc and saturated NH4Cl aqueous solution, and the layers were separated. The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography to provide the title compound. 1 H NMR (600MHz, CDCl3) δ4.12-3.74(m, 3H), 3.18(br s, 1H), 2.66 (dd, J=16.3, 4.6Hz, 1H), 2.33 (dd, J=16.1, 10.5Hz, 1H), 1.49 (s, 9H), 1.32-1.24 (m, 1H), 0.63 (tt, J=8.7, 4.7Hz, 1H), 0.55 (dd, J=12.9, 5.3Hz, 2H), 0.25 (s, 1H), 0.18-0.11 (m, 1H).

[0234] Intermediate A2-2

[0235]

[0236] 3-(difluoromethyl)-5-oxoperidin-1-carboxylic acid tert-butyl ester

[0237] tert-butyl hydroperoxide (1.70 mL, 12.6 mmol, 70% wt / v in H2O) was added dropwise to a solution of tert-butyl 3-oxo-3,6-dihydropyridine-1(2H)-carboxylate (1.1 g, 5.6 mmol) and zinc difluoromethanesulfinate (2.5 g, 8.4 mmol) in trifluorotoluene (22 mL) and H2O (9 mL) at room temperature. The reaction mixture was heated to 35 °C and stirred for 12 h. The flask was then cooled to room temperature, diluted with H2O and DCM, and the layers were separated. The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography (EtOAc: hexane) to provide the title compound. 1 H NMR (600MHz, CDCl3) δ5.80 (t, J=55.5Hz, 1H), 4.12-3.72 (m, 3H), 3.50 (br s, 1H), 2.63 (dd, J=15.7, 5.3Hz, 1H), 2.59-2.53 (m, 1H), 2.50 (dd, J=15.6, 8.7Hz, 1H), 1.47 (s, 9H).

[0238] Intermediate A2-3

[0239]

[0240] tert-butyl 3-oxo-5-(trifluoromethyl)piperidine-1-carboxylate

[0241] To a solution of tert-butyl 3-hydroxy-5-(trifluoromethyl)piperidine-1-carboxylate (8.0 g, 29.7 mmol) in DCM (50 mL), NaHCO3 (7.5 g, 89 mmol) and Desmond oxidant (15.1 g, 35.7 mmol) were added at room temperature. The reaction mixture was stirred at room temperature for 1 h. The reaction was then quenched with H2O and diluted with DCM. The layers were separated and the aqueous phase was extracted with DCM. The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography (EtOAc: hexane) to provide the title compound. 1H NMR (600MHz, CDCl3) δ4.32-3.77 (m, 3H), 3.72-3.20 (m, 1H), 2.83 (d, J=6.9Hz, 1H), 2.73 (dd, J=16.7, 6.1Hz, 1H), 2.57 (dd, J=16.6, 9.4Hz, 1H), 1.47 (s, 9H).

[0242] Intermediate A6-1

[0243]

[0244] 6-Chloro-5-fluoro-5′,5′-dimethylspiro[benzo[d][1,3]oxazine-4,3′-piperidine]-2(1H)-one

[0245] Step 1: 6-Chloro-5-fluoro-5′,5′-dimethyl-2-oxo-1,2-dihydrospiro[benzo[d][1,3]oxazine-4,3′- Piperidine-1′-tert-butyl carboxylate: Under a nitrogen atmosphere, 55 mL of THF was added to a round-bottom flask containing 2.21 g (9.0 mmol) of tert-butyl (4-chloro-3-fluorophenyl)carbamate, and the solution was cooled to -78 °C. The solution was then stirred and added to the mixture over a period of 40 minutes. n BuLi (11.2 mL, 27.9 mmol, 2.5 M in hexane). The reaction mixture was stirred at -78 °C for another 45 minutes, during which time, LaCl3·2LiCl solution (22.5 mL, 13.5 mmol, 0.6 M in THF) and tert-butyl 3,3-dimethyl-5-oxopiperidin-1-carboxylate (3.10 g, 13.5 mmol) were added to the reaction mixture at -78 °C for 40 minutes. The reaction mixture was warmed to room temperature and stirred for 16 h. KO t Bu (5.3 mL, 9.0 mmol, 1.7 M in THF) was added to the reaction mixture, and the solution was heated to 60 °C and maintained for another 3 h. The reaction was cooled to room temperature, quenched with 1 M HCl, and diluted with EtOAc. The layers were separated, and the aqueous phase was extracted with EtOAc. The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography (EtOAc: hexane) to provide the title compound. LCMS [M+Na] + =421.1 (calculated value 421.1).

[0246] Step 2: 6-Chloro-5-fluoro-5′,5′-dimethylspiro[benzo[d]

[13] oxazine-4,3′-piperidine]-2(1H)-one:HCl (25.0 mL, 100 mmol, 4 M in dioxane) was added to a round-bottom flask containing a suspension of tert-butyl 6-chloro-5-fluoro-5′,5′-dimethyl-2-oxo-1,2-dihydrospiro[benzo[d][1,3]oxazine-4,3′-piperidine]-1′-carboxylate (7.98 g, 20.0 mmol) in 30 mL of 1,4-dioxane. The reaction mixture was heated to 90 °C and stirred vigorously for 12 h. The reaction was cooled to room temperature and concentrated to produce the crude title compound, which was used in the next step without further purification. LCMS[M+H] + =299.1 (calculated value 299.1).

[0247] Table A. The following compounds were prepared using appropriate starting materials and a procedure similar to that described with respect to intermediate A6-1.

[0248]

[0249]

[0250] Intermediate A6-15

[0251]

[0252] (rac)-(4R or S, 5′R or S)-6-chloro-5-fluoro-5′-hydroxy-2-oxo-1,2-dihydrospiro[benzo[d][1,3]) [Oxazine-4,3′-piperidine]-1′-tert-butyl carboxylate

[0253] Step 1: 3-((tert-butyldimethylsilyl)oxy)-5-oxopiperidin-1-carboxylic acid tert-butyl ester: A flask containing a solution of tert-butyl 3-hydroxy-5-oxopiperidin-1-carboxylate (22.4 g, 104 mmol) in DMF (224 mL) was cooled to 0 °C. Imidazole (21.2 g, 312 mmol) and TBSCl (18.8 g, 125 mmol) were added, and the reaction mixture was warmed to room temperature and stirred for 16 h. The reaction was quenched with H₂O and extracted with MTBE. The layers were separated, and the combined organic layers were washed with brine. The organic layers were dried over Na₂SO₄, filtered, and concentrated to provide a crude residue, which was purified by silica gel chromatography (EtOAc: petroleum ether) to provide the title compound. LCMS [M-55] + =274.3 (calculated value 274.2).

[0254] Step 2: (rac)-(4R or S, 5′R or S)-5′-((tert-butyldimethylsilyl)oxy)-6-chloro-5-fluoro- 2-Oxo-1,2-dihydrospiro[benzo[d][1,3]oxazine-4,3′-piperidine]-1′-tert-butyl carboxylate and (rac)-(4S or R,5′S) Or R)-5′-((tert-butyldimethylsilyl)oxy)-6-chloro-5-fluoro-2-oxo-1,2-dihydrospiro[benzo[d][1,3] [Oxazine-4,3′-piperidine]-1′-tert-butyl carboxylate: Add 188 mL of THF to a round-bottom flask containing 12.5 g (50.9 mmol) of tert-butyl (4-chloro-3-fluorophenyl)carbamate, and cool the mixture to -78 °C. Add the solution over 1 hour. nBuLi (63.1 mL, 158 mmol, 2.5 M in hexane) was followed by dropwise addition of LaCl3·2LiCl solution (x 102 mL, 61.1 mmol, 0.6 M in THF) and tert-butyl 3-((tert-butyldimethylsilyl)oxy)-5-oxopiperidin-1-carboxylate (20.1 g, 61.1 mmol) over 5 min at -78 °C. The reaction mixture was stirred at -78 °C for 1 h and then warmed to room temperature for another 12 h. The reaction was quenched with a saturated aqueous solution of NH4Cl, poured into an ice-filled flask, and stirred for 15 min. The mixture was diluted with EtOAc, the layers were separated, and the combined organic layers were dried over MgSO4, filtered, and concentrated to provide a crude residue, which was purified by silica gel chromatography (EtOAc: petroleum ether) to provide the title compound as a mixture of diastereomers. The diastereomers were separated by preparative reversed-phase HPLC (ACN / water + 10 mM NH4HCO3). The diastereomer of the title compound was obtained by rapid elution: LCMS[M+Na] + = 523.3 (calculated value 523.2). The slower-eluting diastereomer of the title compound is obtained: LCMS[M+Na] + =523.3 (calculated value 523.2).

[0255] Step 3: (rac)-(4R or S, 5′R or S)-6-chloro-5-fluoro-5′-hydroxyspiro[benzo[d][1,3]oxazine-4,3′- Piperidine-2(1H)-one: The flask containing (rac)-(4S or R, 5′S or R)-5′-((tert-butyldimethylsilyl)oxy)-6-chloro-5-fluoro-2-oxo-1,2-dihydrospiro[benzo[d][1,3]oxazine-4,3′-piperidine]-1′-carboxylic acid tert-butyl ester (from the slower elution peak of the previous step, 1.0 g, 2.0 mmol) in THF (20 mL) was cooled to 0 °C. TBAF (6.0 mL, 6.0 mmol) was added, and the reaction mixture was warmed to 40 °C and maintained for 12 h. The reaction was quenched with ice water and stirred for 15 min. The mixture was diluted with EtOAc, the layers were separated, and the combined organic layers were washed with brine. The organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure to provide a crude product, which was used without further purification. HCl (0.9 mL, 3.5 mmol, 4 M in dioxane) was added to a tubular flask containing a suspension of the crude product (34 mg, 0.090 mmol) in 1,4-dioxane (0.8 mL). The reaction mixture was stirred vigorously at room temperature for 12 h and concentrated to produce the crude title compound. The crude product was used for the next step without further purification. LCMS [M+H] + =287.0 (calculated value 287.1).

[0256] Intermediate A6-16

[0257]

[0258] (rac)-(4R or S, 5′R or S)-6-chloro-5,5′-difluorospiro[benzo[d][1,3]oxazine-4,3′-piperidine]-2 (1H)-keto

[0259] Step 1: (rac)-(4R or S, 5′R or S)-6-chloro-5,5′-difluoro-2-oxo-1,2-dihydrospiro[benzo[d]] [1,3]oxazine-4,3′-piperidine]-1′-tert-butyl carboxylate: A flask containing (rac)-(4S or R, 5′S or R)-5′-((tert-butyldimethylsilyl)oxy)-6-chloro-5-fluoro-2-oxo-1,2-dihydrospiro[benzo[d][1,3]oxazine-4,3′-piperidine]-1′-carboxylic acid tert-butyl ester (from the slower elution peak of step 1 from intermediate A6-15, 1.0 g, 2.0 mmol) in THF (20 mL) was cooled to 0 °C. TBAF (6.0 mL, 6.0 mmol, 1 M THF solution) was added to the flask, and the reaction mixture was warmed to 40 °C and maintained for 12 h. The reaction was quenched with ice water and stirred for 15 min. The mixture was diluted with EtOAc, the layers were separated, and the combined organic layers were washed with brine. The organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure to provide a crude product, which was used without further purification. Crude ethanol (100 mg, 0.260 mmol) in DCM (5 mL) was cooled to -78 °C under a nitrogen atmosphere. DAST (625 mg, 0.390 mmol) was added dropwise to DCM (2 mL), and the reaction mixture was stirred at -78 °C for 1 h. The reaction mixture was then purified directly by silica gel chromatography (EtOAc:DCM) to provide the title compound. LCMS [M+H] + =411.3 (calculated value 411.1).

[0260] Step 2: (rac)-(4R or S, 5′R or S)-6-chloro-5,5′-difluorospiro[benzo[d][1,3]oxazine-4,3′-piperazine [Pyridine]-2(1H)-one: HCl (0.6 mL, 2.3 mmol, 4 M in dioxane) was added to a tubular flask containing (rac)-(4R or S, 5′R or S)-6-chloro-5,5′-difluoro-2-oxo-1,2-dihydrospiro[benzo[d][1,3]oxazine-4,3′-piperidine]-1′-carboxylic acid tert-butyl ester (45 mg, 0.12 mmol) in 1,4-dioxane (0.6 mL), and the reaction mixture was stirred at room temperature for 3 h. The solvent was removed under reduced pressure to produce a crude product, which was used for the next step without further purification. LCMS [M+H] + =289.1 (calculated value 289.1).

[0261] Intermediate A6-17

[0262]

[0263] (4R and S, 6′S)-6-chloro-5-fluoro-6′-methylspiro[benzo[d][1,3]oxazine-4,3′-piperidine]-2(1H)-one

[0264] Step 1: (4R and S, 6′S)-6-chloro-5-fluoro-6′-methyl-2-oxo-1,2-dihydrospiro[benzo[d][1,3]ox [4,3′-piperidine]-1′-carboxylic acid benzyl ester: Add 15 mL of THF to 600 mg (2.44 mmol) of tert-butyl (4-chloro-3-fluorophenyl)carbamate and cool to -78 °C. The reaction time is 40 min. n BuLi (3.0 mL, 7.57 mmol, 2.5 M in hexane) was added, and the resulting mixture was stirred at -78 °C for another 45 min. LaCl3·2LiCl solution (6.1 mL, 3.66 mmol, 0.6 M in THF) and (S)-2-methyl-5-oxopiperidin-1-carboxylic acid benzyl ester (900 mg, 3.66 mmol) were added at -78 °C for 40 min, and the reaction mixture was warmed to room temperature and stirred for 16 h. KO was then added. t Bu (1.40 mL, 2.44 mmol, 1.7 M in THF) was added, and the reaction was heated to 60 °C and maintained for another 3 h. The reaction was cooled to room temperature, quenched with 1 M HCl, and diluted with EtOAc. The layers were separated, and the aqueous layer was extracted with EtOAc. The combined organic layers were dried over MgSO4, filtered, and concentrated to provide a crude residue, which was purified by silica gel chromatography (EtOAc: hexane) to provide the title compound as a mixture of diastereomers. LCMS [M+H] + =419.0 (calculated value 419.1).

[0265] Step 2: (4R and S, 6′S)-6-chloro-5-fluoro-6′-methylspiro[benzo[d][1,3]oxazine-4,3′-piperidine]-2 (1H)-Ketone: HBr (3.9 mL, 71.6 mmol, 33 wt% in AcOH) was added to a tubular flask containing (4R and S, 6′S)-6-chloro-5-fluoro-6′-methyl-2-oxo-1,2-dihydrospiro[benzo[d][1,3]oxazine-4,3′-piperidine]-1′-carboxylate (600 mg, 1.43 mmol). The reaction mixture was heated to 80 °C and maintained for 12 h. The reaction was cooled to room temperature and concentrated to produce the crude title compound, which was used for the next step without further purification. LCMS [M+H] + =285.1 (calculated value 285.1).

[0266] Intermediate B4-1

[0267]

[0268] rac-(1,1,1-trifluorobutane-2-yl)hydrazine

[0269] Step 1: N′-(1,1,1-trifluorobutane-2-ethylenedimethylbenzylhydrazine):Benzoylhydrazine (1.50 g, 11.0 mmol) was added to a solution of 1,1,1-trifluorobutane-2-one (1.39 g, 11.0 mmol) in toluene (10 mL), and the reaction mixture was heated to 110 °C and maintained for 18 h. The reaction was cooled to room temperature, poured into water, and then filtered. The solid was washed with water and further dried to yield the desired crude title compound. LCMS[M+H] + =245.4 (calculated value 245.1).

[0270] Step 2: N′-(1,1,1-trifluorobutane-2-yl)benzoylhydrazine: A solution of N′-(1,1,1-trifluorobutane-2-yl)benzoylhydrazine (500 mg, 2.05 mmol) in THF (6 mL) was cooled to 0 °C and BH3 THF (4.09 mL, 4.09 mmol, 1.0 M THF solution) was added dropwise. The reaction was warmed to room temperature and stirred for 14 h. The reaction was then recooled to 0 °C and quenched with MeOH. The solvent was removed under reduced pressure and dichloromethane was added. The slurry was filtered to remove insoluble matter, and the organic layer was washed with a saturated aqueous solution of NH4Cl, dried over MgSO4, filtered, and concentrated to provide the title compound. LCMS [M+H] + =246.7 (calculated value 247.1).

[0271] Step 3: (1,1,1-trifluorobutane-2-yl)hydrazine: Hydrogen chloride (1.48 mL, 17.8 mmol, 37% aqueous solution) was added to a solution of N′-(1,1,1-trifluorobutan-2-yl)benzoylhydrazine (274 mg, 1.11 mmol) in MeOH (3 mL), and the resulting mixture was heated to 80 °C and maintained for 16 h. The reaction was cooled to room temperature and concentrated under reduced pressure. EtOAc was added, and the precipitate was filtered and washed with EtOAc to provide the title compound. 1 H NMR (400MHz, CD3OD) δ3.34-3.49 (m, 1H), 1.80 (dqd, J=14.7, 7.5, 4.4Hz, 1H), 1.46-1.65 (m, 1H), 0.97-1.27 (m, 3H).

[0272] Table B. Prepare the following compounds using appropriate starting materials and procedures similar to those described in Intermediate B4-1.

[0273]

[0274]

[0275] Intermediate C4-1

[0276]

[0277] (2,2,2-trifluoro-1-(1-fluorocyclopropyl)ethyl)hydrazine

[0278] Step 1: N′-((1-fluorocyclopropyl)methylene)benzoylhydrazine: 1-Fluorocyclopropane-1-carboxaldehyde (176 mg, 2.0 mmol) was added to a solution of benzoylhydrazine (272 mg, 2.0 mmol) in toluene (4 mL), and the reaction mixture was heated to 60 °C and maintained for 1 h. The reaction was cooled to room temperature and concentrated to provide a crude product, which was used in the next step without any purification. LCMS [M+H] + =207.1 (calculated value 207.1).

[0279] Step 2: N′-(2,2,2-trifluoro-1-(1-fluorocyclopropyl)ethyl)benzoylhydrazine: Allyltrimethylsilane (0.48 mL, 3.0 mmol) and BF3·Et2O (0.37 mL, 3.0 mmol) were added sequentially to a suspension of N′-((1-fluorocyclopropyl)methylene)benzoylhydrazine (412 mg, 2.0 mmol) in 1,2-dichloroethane (4.0 mL), and the mixture was refluxed for 5 min. The solvent was evaporated under vacuum, and the resulting residue was dissolved in DMF (4 mL). TMSCF3 (0.60 mL, 4.0 mmol) and NaOAc (660 mg, 8.0 mmol) were added, and the mixture was heated to 55 °C and maintained for 3 h. The reaction was cooled to room temperature and quenched with a saturated aqueous solution of Na2CO3, and stirred for another 5 min. The mixture was diluted with H2O and extracted with EtOAc. The combined organic layers were dried over MgSO4, filtered, and concentrated to provide a crude residue, which was purified by silica gel chromatography (EtOAc: hexane) to provide the title compound. LCMS[M+H] + =277.1 (calculated value 277.1).

[0280] Step 3: (2,2,2-trifluoro-1-(1-fluorocyclopropyl)ethyl)hydrazine: To a solution of N′-(2,2,2-trifluoro-1-(1-fluorocyclopropyl)ethyl)benzoylhydrazine (140 mg, 0.51 mmol) in MeOH (0.75 mL), HCl (0.7 mL, 8.1 mmol, 37% aqueous solution) was added, and the resulting mixture was heated to 80 °C and maintained for 16 h. The reaction was cooled to room temperature and concentrated to provide the crude product, which was azeotropically reacted with toluene to provide the title compound, which was then used without purification. LCMS[M+H] + =173.1 (calculated value 173.1).

[0281] Table C. Prepare the following compounds using appropriate starting materials and procedures similar to those described in intermediate C4-1.

[0282]

[0283] Intermediate D2-1

[0284]

[0285] ((1-Fluorocyclopropyl)methyl)hydrazine

[0286] Hydrazine (0.33 mL, 0.33 mmol, 1.0 M THF solution) was added to a tubular flask containing 1-(bromomethyl)-1-fluorocyclopropane (50 mg, 0.33 mmol) in EtOH (0.3 mL), and the resulting mixture was heated to 70 °C and maintained for 16 h. The reaction was cooled to room temperature and concentrated to provide the crude product, which was then azeotropically reacted with toluene to provide the title compound, which was used without purification. LCMS [M+H] + =105.1 (calculated value 105.1).

[0287] Table D. Prepare the following compounds using appropriate starting materials and procedures similar to those described in Intermediate D2-1.

[0288]

[0289] Intermediate E3-1

[0290]

[0291] (1-(4-fluorophenyl)cyclopropyl)hydrazine

[0292] Step 1: Di-tert-butyl 1-(1-(4-fluorophenyl)cyclopropyl)hydrazine-1,2-dicarboxylate: Add acetonitrile (20 mL) to a mixture containing 1-(4-fluorophenyl)cyclopropane-1-carboxylic acid (360 mg, 2.0 mmol) and [Mes-Acr-Me]. + The photocatalyst (16 mg, 0.04 mmol) was placed in a tubular flask. The solution was degassed with N2 for 5 min. After degassed, DBU (0.06 mL, 0.4 mmol) and di-tert-butyl azodicarbonate (576 mg, 2.5 mmol) were rapidly and continuously added. The tubular flask was placed in front of a 450 nm blue LED (Merck photoreactor) and stirred for 12 h. The solvent was removed under reduced pressure, and the crude mixture was purified by silica gel chromatography (EtOAc: hexane) to provide the title compound. LCMS [M-155] + =211.1 (calculated value 211.2).

[0293] Step 2: (1-(4-fluorophenyl)cyclopropyl)hydrazine:HCl (6.6 mL, 26.2 mmol, 4.0 M dioxane solution) was added to a tubular flask containing di-tert-butyl 1-(1-(4-fluorophenyl)cyclopropyl)hydrazine-1,2-dicarboxylate (640 mg, 1.75 mmol), and the reaction mixture was stirred at room temperature for 16 h. The solvent was removed under reduced pressure, and the mixture was azeotropically reacted with toluene to provide the title compound, which was then used without purification. LCMS [M+H] + =167.0 (calculated value 167.1).

[0294] Table E. Prepare the following compounds using appropriate starting materials and procedures similar to those described in Intermediate E3-1.

[0295]

[0296] Intermediate F4-1

[0297]

[0298] (1-(trifluoromethyl)cyclopropyl)hydrazine

[0299] Step 1: (1-(trifluoromethyl)cyclopropyl)tert-butyl carbamate: 1-(trifluoromethyl)cyclopropanecarboxylic acid (5.00 g, 32.4 mmol) was subjected to... t TEA (5.00 mL, 35.7 mmol) and diphenylphosphinyl azide (11.8 g, 48.7 mmol) were added to a solution of BuOH (5 mL), and the resulting mixture was stirred at room temperature for 0.5 h and then heated to 100 °C and held for 15 h. The reaction mixture was diluted with EtOAc and washed with 5% citric acid, saturated NaHCO3 aqueous solution, and brine. The organic layer was dried over Na2SO4, filtered, and concentrated to provide a crude product, which was purified by silica gel chromatography (EtOAc: petroleum ether) to yield the title compound. 1 H NMR (400MHz, CDCl3) δ4.99-5.12 (m, 1H), 1.45 (s, 9H), 1.26 (br s, 2H), 1.11 (br s, 2H).

[0300] Step 2: tert-butyl nitroso(1-(trifluoromethyl)cyclopropyl)carbamate: Nitros(tetrafluoroborate) (78 mg, 0.67 mmol) was aliquoted into a solution of tert-butyl (100 mg, 0.440 mmol) of pyridine (0.2 mL) and acetonitrile (2 mL) at -30 °C. The solution was stirred at -30 °C for 30 min, then warmed to 0 °C and held for 2 h. The reaction was concentrated under reduced pressure to produce a crude product, which was purified by preparative TLC (EtOAc: petroleum ether) to provide the title compound.1 H NMR (400MHz, CDCl3) δ1.69 (s, 9H), 1.57-1.62 (m, 4H).

[0301] Step 3: (1-(trifluoromethyl)cyclopropyl)hydrazine: 100 mg (0.390 mmol) of tert-butyl nitroso(1-(trifluoromethyl)cyclopropyl)carbamate was stirred in MeOH (2 mL) at -78 °C for 30 min. HCl (0.32 mL, 3.9 mmol, 37% aqueous solution) and zinc (257 mg, 3.93 mmol) were added at -78 °C, and the resulting mixture was stirred for 2 h. The reaction was warmed to room temperature, filtered, and concentrated to provide the title compound, which was then used without purification. LCMS [M+H] + =141.0 (calculated value 141.1).

[0302] Intermediate G2-1

[0303]

[0304] (2-(difluoromethoxy)phenyl)hydrazine

[0305] Add THF (1 ml) to NaO t A mixture of Bu (129 mg, 1.35 mmol) and X-PHOS Pd G2 (10.6 mg, 0.0130 mmol) was added. 1-Bromo-2-(difluoromethoxy)benzene (300 mg, 1.35 mmol) was added, and the mixture was stirred at room temperature for 10 min. Hydrazine (42 μL, 1.3 mmol) was added in a single addition, and the tubular flask was heated to 90 °C (preheated bath) and stirred for 12 h. The reaction was cooled to room temperature, diluted with MeOH, and filtered. The filtrate was concentrated to dryness, and the crude mixture was purified by reversed-phase preparative HPLC (C18 stationary phase, ACN / water + 0.1% TFA) to provide the title compound. LCMS [M+H] + =175.0 (calculated value 175.1).

[0306] Table G. Prepare the following compounds using appropriate starting materials and procedures similar to those described in Intermediate G2-1.

[0307]

[0308]

[0309] Intermediate H4-1

[0310]

[0311] 5-Amino-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-pyrazol-4-carboxylic acid

[0312] Step 1: Ethyl 5-amino-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-pyrazole-4-carboxylate: Sodium hydride (30.9 mg, 1.29 mmol) was added to a stirred solution of ethyl 5-amino-1H-pyrazole-4-carboxylate (100 mg, 0.650 mmol) in 1 mL of ACN at 0 °C. After 1 h, 4-(bromomethyl)tetrahydro-2H-pyran (173 mg, 0.970 mmol) was added, and the resulting mixture was heated to 80 °C and held for 12 h. The reaction was cooled to room temperature and quenched with a saturated aqueous solution of NH4Cl. The aqueous phase was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by preparative reversed-phase HPLC (C18 stationary phase, ACN / water + 0.04% NH4OH) to provide the title compound. LCMS [M+H] + =254.2 Anhydrous (Calculated value 254.1).

[0313] Step 2: 5-Amino-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-pyrazol-4-carboxylic acid: LiOH·H₂O (20 mg, 0.47 mmol) was added to a solution of ethyl 5-amino-1-((tetrahydro-2H-pyran-4-yl)methyl)-1H-pyrazole-4-carboxylate (100 mg, 0.400 mmol) in EtOH (1 mL) and H₂O (0.2 mL), and the resulting mixture was heated to 60 °C and maintained for 12 h. The mixture was concentrated to produce a crude product, which was used in the next step without any purification. LCMS[M+H] + =226.0 (calculated value 226.1).

[0314] Table H. Prepare the following compounds using appropriate starting materials and procedures similar to those described in Intermediate H4-1.

[0315]

[0316] Intermediate I4-1

[0317]

[0318] 5-Amino-1-(1-(trifluoromethyl)cyclopropyl)-1H-pyrazole-4-carboxylic acid

[0319] Step 1: Ethyl 5-amino-1-(1-(trifluoromethyl)cyclopropyl)-1H-pyrazole-4-carboxylate: A solution of (1-(trifluoromethyl)cyclopropyl)hydrazine (50 mg, 0.36 mmol), methyl 2-cyano-3-ethoxyacrylate (56 mg, 0.36 mmol), and DIEA (0.31 mL, 1.8 mmol) in EtOH (1 mL) was heated to 80 °C and maintained for 12 h. The reaction was cooled to room temperature and concentrated under reduced pressure to produce a crude product, which was purified by preparative TLC (EtOAc: petroleum ether) to provide the title compound. LCMS [M+H] +=264.1 (calculated value 264.1).

[0320] Step 2: 5-Amino-1-(1-(trifluoromethyl)cyclopropyl)-1H-pyrazole-4-carboxylic acid: LiOH·H₂O (42.1 mg, 1.0 mmol) was added to a solution of ethyl 5-amino-1-(1-(trifluoromethyl)cyclopropyl)-1H-pyrazole-4-carboxylate (50 mg, 0.20 mmol) in MeOH (2 mL) and H₂O (0.5 mL). The resulting mixture was heated to 70 °C and maintained for 5 h. The reaction was cooled to room temperature, concentrated, acidified to pH 2 with 1 M HCl, and extracted with EtOAc. The combined organic layers were dried over Na₂SO₄, filtered, and concentrated under reduced pressure to produce a crude product, which was used without further purification. LCMS[M+H] + =235.9 (calculated value 236.1).

[0321] Intermediate I4-2

[0322]

[0323] Intermediate I4-2 was prepared using a procedure similar to that described above for intermediate I4-1. LCMS[M+H] + =210.1 (calculated value 210.0).

[0324] Intermediate L3-1

[0325]

[0326] (R,E / Z)-2-(6-chloro-5-fluoro-2-oxo-1,2-dihydrospiro[benzo[d][1,3]oxazine-4,3′-piperidine]- 1′-carbonyl)-3-ethoxyacrylonitrile

[0327] Step 1: 3-(6-chloro-5-fluoro-2-oxo-1,2-dihydrospiro[benzo[d][1,3]oxazine-4,3′-piperidine]-1′- 3-Oxypropionitrile: DIEA (7.06 mL, 40.4 mmol) was added to a solution of 6-chloro-5-fluoro-2-oxo-1,2-dihydrospiro[benzo[d][1,3]oxazine-4,3′-piperidine]-1′-onium chloride (4.14 g, 13.5 mmol) and 2-cyanoacetic acid (1.26 g, 14.8 mmol) in EtOAc (41 mL) and DMF (6.2 mL). 1-Propanephonic anhydride (9.60 mL, 16.2 mmol) was added, and the resulting mixture was stirred overnight at room temperature. The reaction was quenched with H₂O and extracted with EtOAc. The combined organic layers were washed with brine, dried over MgSO₄, and concentrated to provide a crude residue, which was purified by silica gel chromatography ((3:1)EtOH:EtOAc):hexane) to provide the title compound. LCMS [M+H] + =337.9 (calculated value 338.1).

[0328] Step 2: (R,E / Z)-2-(6-chloro-5-fluoro-2-oxo-1,2-dihydrospiro[benzo[d][1,3]oxazine-4,3′-piperazine) [Pyridine]-1′-carbonyl)-3-ethoxyacrylonitrile: ZnCl2 (anhydrous, 303 mg, 2.21 mmol) was added to 3-(6-chloro-5-fluoro-2-oxo-1,2-dihydrospiro[benzo[d][1,3]oxazine-4,3′-piperidin]-1′-yl)-3-oxopropionitrile (2.50 g, 7.40 mmol) in triethyl orthoformate (18.5 mL, 111 mmol) and NMP (0.8 mL), and the reaction mixture was heated to 130 °C and maintained for 4 h. The reaction was cooled to room temperature, quenched with a saturated aqueous solution of NaHCO3, and extracted with EtOAc. The combined organic layers were dried over MgSO4, filtered, and concentrated to provide a crude residue, which was purified by silica gel chromatography (EtOAc: hexane) to provide the title compound. LCMS [M+H] + =394.0 (calculated value 394.1).

[0329] Intermediate L3-2

[0330]

[0331] (R)-2-(6-chloro-5-fluoro-2-oxo-1,2-dihydrospiro[benzo[d][1,3]oxazine-4,3′-piperidine]-1′-carbonyl) 3-hydroxyacrylonitrile

[0332] THF (15 mL) was added to a mixture of (R)-3-(6-chloro-5-fluoro-2-oxo-1,2-dihydrospiro[benzo[d][1,3]oxazin-4,3′-piperidin]-1′-yl)-3-oxopropionitrile (1.82 g, 4.43 mmol) and methyl formate (3.99 g, 66.5 mmol). The reaction mixture was sonicated to dissolve the solids, and then KO was added dropwise. t The solution was prepared using Bu (14.2 mL, 14.2 mmol, 1 M in THF), and the resulting mixture was stirred at room temperature for 12 h. The reaction mixture was diluted with H₂O, neutralized to pH 6 with 1 M HCl, and extracted with EtOAc. The combined organic layers were dried over MgSO₄, filtered, and concentrated to provide a crude title compound, which was used in the next step without any purification. LCMS [M+H] + =366.4 (calculated value 366.1).

[0333] Example 1

[0334]

[0335] (R)-1′-(5-amino-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carbonyl)-6-chloro-5-fluorospiro[benzo[d]] [1,3]oxazine-4,3′-piperidine]-2(1H)-one

[0336] (R, E / Z)-2-(6-chloro-5-fluoro-2-oxo-1,2-dihydrospiro[benzo[d][1,3]oxazine-4,3′-piperidine]-1′-carbonyl)-3-ethoxyacrylonitrile (0.25 mL, 0.10 mmol, 0.4 M stock solution in EtOH) was added to a tubular flask containing (2,2,2-trifluoroethyl)hydrazine (17 mg, 0.15 mmol). TEA (42 μL, 0.30 mmol) was added to the tubular flask, and the mixture was heated to 70 °C and maintained for 12 h. The reaction was cooled to room temperature and directly purified by preparative reversed-phase HPLC (C18 stationary phase, ACN / water + 0.05% TFA) to provide the title compound. 1 H NMR (500MHz, CD3OD) δ7.55 (s, 1H), 7.43 (t, J=8.1Hz, 1H), 6.74 (d, J=8.6Hz, 1H), 4.76-4.66 (m, 3H), 4.46 (d, J=13.3Hz, 1H), 3. 53 (s, 1H), 3.24-3.16 (m, 1H), 2.52 (t, J=11.5Hz, 1H), 2.30 (d, J=13.6Hz, 1H), 2.17 (q, J=10.4Hz, 1H), 1.73 (d, J=12.7Hz, 1H). LCMS[M+H] + =462.3 (calculated value 462.1).

[0337] Example 2

[0338]

[0339] (R)-1′-(5-amino-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carbonyl)-6-chloro-5-fluoro-5′,5′-di Methylspiro[benzo[d][1,3]oxazine-4,3′-piperidine]-2(1H)-one

[0340] (R)-6-chloro-5-fluoro-5′,5′-dimethylspiro[benzo[d][1,3]oxazine-4,3′-piperidine]-2(1H)-one (67 mg, 0.20 mmol) and 5-amino-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxylic acid (84 mg, 0.40 mmol) were placed in a 3 mL tubular flask containing 0.2 mL of DMF. TEA (0.14 mL, 1.0 mmol) was added to this mixture, followed by T3P (0.18 μL, 0.6 mmol, 50% w / v in DMF), and the resulting mixture was stirred at room temperature for 2 h. The reaction mixture was directly purified by preparative reversed-phase HPLC (C18 stationary phase, ACN / water + 0.05% HCO2H) to provide the title compound. 1H NMR (500MHz, CD3OD) δ7.62 (s, 1H), 7.44-7.40 (m, 1H), 6.74 (dd, J=8.7, 1.1Hz, 1H), 4.79-4.70 (m, 3H), 4. 27 (d, J=12.9Hz, 1H), 3.73 (s, 1H), 2.97 (d, J=12.4Hz, 1H), 2.17-2.03 (m, 2H), 1.24 (s, 3H), 1.03 (s, 3H). LCMS[M+H] + =490.4 (calculated value 490.1).

[0341] Examples 3 and 4

[0342]

[0343] (R)-1′-(5-amino-1-((R)-1,1,1-trifluorobutane-2-yl)-1H-pyrazole-4-carbonyl)-6-chloro-5-fluoro Spiro[benzo[d][1,3]oxazine-4,3′-piperidine]-2(1H)-one and (R)-1′-(5-amino-1-((S)-1,1,1-trifluorobutane) Alkyl)-1H-pyrazole-4-carbonyl)-6-chloro-5-fluorospiro[benzo[d][1,3]oxazine-4,3′-piperidine]-2(1H)-one

[0344] (R)-2-(6-chloro-5-fluoro-2-oxo-1,2-dihydrospiro[benzo[d][1,3]oxazine-4,3′-piperidine]-1′-carbonyl)-3-hydroxyacrylonitrile (0.25 mL, 0.01 mmol, 0.4 M stock solution in AcOH) was added to a tubular flask containing R- and S-(1,1,1-trifluorobutane-2-yl)hydrazine (31 mg, 0.15 mmol), and the resulting mixture was heated to 80 °C and maintained for 12 h. The crude mixture was directly purified by preparative reversed-phase HPLC (C18 stationary phase, ACN / water + 0.05% TFA) to provide a mixture of diastereomers of the title compound. The title compound was resolved by preparative chiral SFC using method A. The faster-eluting isomers of the title compound were obtained (Example 3): 1 H NMR (400MHz, CD3OD) δ7.57 (s, 1H), 7.41 (dd, J=8.6, 7.8Hz, 1H), 6.72 (dd, J=8.7, 1.3Hz, 1H), 4.74-4.81 (m, 1H), 4.68 (br d, J=13.4Hz, 1H), 4.45 (br d, J=12.2Hz, 1H), 3.43-3.58(m, 1H), 3.05-3.25(m, 1H), 2.42-2.58(m, 1H), 2.24-2.34(m, 2H ), 2.11-2.22 (m, 1H), 1.95-2.07 (m, 1H), 1.72 (dt, J=11.3, 2.2Hz, 1H), 0.82 (t, J=7.3Hz, 3H). LCMS[M+H] += 490.1 (calculated value 490.1). A slower-eluting isomer of the title compound was obtained (Example 4). 1 H NMR (400MHz, CD3OD) δ7.57 (s, 1H), 7.41 (dd, J=8.7, 7.7Hz, 1H), 6.73 (dd, J=8.7, 1.3Hz, 1H), 4.76-4.80 (m, 1H), 4.68 (br d, J=14.9Hz, 1H), 4.46 (br d, J=11.5Hz, 1H), 3.41-3.62(m, 1H), 3.02-3.25(m, 1H), 2.45-2.60(m, 1H), 2.22-2.36(m, 2H) , 2.10-2.22 (m, 1H), 2.00 (dqd, J=14.1, 7.3, 3.9Hz, 1H), 1.65-1.77 (m, 1H), 0.80-0.88 (m, 3H). LCMS[M+H] + =490.2 (calculated value 490.1).

[0345] Table 1. The following compounds were prepared using suitable starting materials according to a procedure similar to that described with respect to Examples 1-4.

[0346]

[0347]

[0348]

[0349]

[0350]

[0351]

[0352]

[0353]

[0354]

[0355]

[0356]

[0357]

[0358]

[0359]

[0360]

[0361]

[0362]

[0363]

[0364]

[0365] Table 2. The following compounds were prepared using suitable starting materials according to a procedure similar to that described with respect to Examples 3 and 4. Diasteremeric products were separated using the chiral SFC method specified in the table. For those diastereomer pairs, the rapidly eluted isomer is listed first.

[0366]

[0367]

[0368]

[0369]

[0370]

[0371] Example 132

[0372]

[0373] (R)-6-chloro-5-fluoro-1′-(1-(4-fluorobenzyl)-5-hydroxy-1H-pyrazole-4-carbonyl)spiro[benzo[d][1,3] [Oxazine-4,3′-piperidine]-2(1H)-one

[0374] Step 1: Ethyl 1-(4-fluorobenzyl)-5-hydroxy-1H-pyrazole-4-carboxylate: K₂CO₃ (1.27 g, 9.20 mmol) was added to a solution of (4-fluorobenzyl)hydrazine hydrochloride (650 mg, 3.68 mmol) and diethyl 2-(ethoxymethylene)malonate (875 mg, 4.05 mmol) in H₂O (3 mL), and the resulting mixture was heated to 100 °C and maintained for 3 h. The reaction was cooled to room temperature, and the mixture was washed with EtOAc. The aqueous phase was acidified to pH 2 with 1 M HCl and extracted with EtOAc. The organic layer was dried over Na₂SO₄, filtered, and concentrated under reduced pressure to produce the crude title compound, which was used without further purification. LCMS [M+H] + =265.0 (calculated value 265.1).

[0375] Step 2: 1-(4-fluorobenzyl)-5-hydroxy-1H-pyrazole-4-carboxylic acid:To a solution of ethyl 1-(4-fluorobenzyl)-5-hydroxy-1H-pyrazole-4-carboxylate (25 mg, 0.095 mmol) in EtOH (1 mL) and H₂O (0.2 mL), NaOH (38 mg, 0.95 mmol) was added, and the resulting mixture was heated to 90 °C and maintained for 4 h. The reaction was cooled to room temperature and concentrated to provide a crude residue, which was acidified to pH 2 with 1 M HCl and extracted with EtOAc. The organic layer was subjected to Na₂SO₄... 4- The compound was dried, filtered, and concentrated under reduced pressure to produce a crude title compound, which was then used without further purification. LCMS[M+H] + =237.0 (calculated value 237.1).

[0376] Step 3: (R)-6-chloro-5-fluoro-1′-(1-(4-fluorobenzyl)-5-hydroxy-1H-pyrazole-4-carbonyl)spiro[benzo[d] [1,3]oxazine-4,3′-piperidine]-2(1H)-one: TCFH (26 mg, 0.093 mmol) and 1-methylimidazole (21 mg, 0.25 mmol) were added to a solution of 1-(4-fluorobenzyl)-5-hydroxy-1H-pyrazole-4-carboxylic acid (20 mg, 0.085 mmol) and (R)-6-chloro-5-fluorospiro[benzo[d][1,3]oxazine-4,3′-piperidine]-2(1H)-one (31 mg, 0.085 mmol) in ACN (2 mL), and the resulting mixture was stirred at room temperature for 12 h. The reaction was concentrated to provide a crude residue, which was purified by preparative reversed-phase HPLC (C18 stationary phase, ACN / water + 0.1% TFA) to provide a mixture of diastereomers of the title compound. 1 H NMR (500MHz, CD3OD) δ7.91 (br s, 1H), 7.46 (t, J=8.2Hz, 1H), 7.32 (br s, 2H), 7.09 (br t, J=8.5Hz, 2H), 6.76 (br d, J=8.4Hz, 1H), 5.13 (br s, 2H), 4.36 (s, 2H), 3.19 (br s, 2H), 2.52 (br s, 1H), 2.31 (br d, J=13.7Hz, 1H), 2.17 (br d, J=13.1Hz, 1H), 1.75 (br d, J=14.5Hz, 1H). LCMS[M+H] + =489.1 (calculated value 489.1).

[0377] Example 133

[0378]

[0379] (R)-1′-(5-amino-1-benzyl-1H-1,2,3-triazol-4-carbonyl)-6-chloro-5-fluorospiro[benzo[d][1,3]) [Oxazine-4,3′-piperidine]-2(1H)-one

[0380] (R)-6-chloro-5-fluoro-2-oxo-1,2-dihydrospiro[benzo[d][1,3]oxazine-4,3′-piperidine]-1′-onium 2,2,2-trifluoroacetate (50 mg, 0.13 mmol) and 5-amino-1-benzyl-1H-1,2,3-triazol-4-carboxylic acid (28 mg, 0.13 mmol) were added to the mixture. DIEA (23 μL, 0.13 mmol) was then added in a single dose, followed by HATU (49 mg, 0.13 mmol), and the mixture was stirred at room temperature for 2 h. The reaction mixture was directly purified by preparative reversed-phase HPLC (C18 stationary phase, ACN / water + 0.05% TFA) to provide the title compound. 1 H NMR (500MHz, CD3OD) δ7.91 (br s, 1H), 7.46 (t, J=8.2Hz, 1H), 7.32 (br s, 2H), 7.09 (brt, J=8.5Hz, 2H), 6.76 (br d, J=8.4Hz, 1H), 5.13 (br s, 2H), 4.36 (s, 2H), 3.19 (br s, 2H), 2.52 (br s, 1H), 2.31 (br d, J=13.7Hz, 1H), 2.17 (br d, J=13.1Hz, 1H), 1.75 (br d, J=14.5Hz, 1H). LCMS[M+H] + =489.1 (calculated value 489.1).

[0381] Factor XIa determination

[0382] The efficacy of the compounds of the present invention as inhibitors of coagulation factor XIa can be determined using the relevant purified serine protease and appropriate synthetic substrates. The hydrolysis rate of the relevant serine protease on a chromogenic or fluorescent substrate was measured in the absence and presence of the compounds of the present invention. The determination was performed at room temperature or 37°C. The hydrolysis of the substrate resulted in the release of aminotrifluoromethylcoumarin (AFC), which was monitored by fluorescence spectroscopy by measuring the increase in emission at 510 nm under 405 nm excitation. A decrease in the rate of fluorescence change in the presence of the inhibitor indicates enzyme inhibition. Such methods are known to those skilled in the art. The results of this determination are expressed as the half-maximal inhibitory concentration (IC50). 50 or suppression constant K i .

[0383] The compound was incubated with human (0.04 nM) factor XIa at 25 °C for 30 min in 50 mM HEPES buffer containing 150 mM sodium chloride, 5 mM calcium chloride, and 0.1% PEG8000 at pH 7.4. Factor XIa enzyme activity was determined by fluorescence measurement at 400 / 505 nm after adding the substrate glycine-proline-arginine-7-acylamino-4-trifluoromethylcoumarin (GPR-AFC) and incubating at 25 °C for 60 min. The percentage of inhibition at each data point was calculated from the data, and analysis was performed using a four-parameter equation of log(inhibitor) versus response to determine the half-maximum inhibitory concentration (IC50). 50 Using the Cheng-Prusoff equation to convert IC 50 It is converted into the equilibrium inhibition constant (Ki).

[0384] The activity indicated by this assay suggests that the compounds of the present invention can be used therapeutically to treat or prevent various cardiovascular and / or cerebrovascular thromboembolic conditions in patients suffering from unstable angina, acute coronary syndrome, refractory angina, myocardial infarction, transient ischemic attack, atrial fibrillation, stroke such as thrombotic or embolic stroke, venous thrombosis, coronary and cerebral artery thrombosis, cerebral and pulmonary embolism, atherosclerosis, deep vein thrombosis, disseminated intravascular coagulation, and re-occlusion or restenosis of recanalized vessels.

[0385] Plasma kallikrein assay

[0386] The effectiveness of the compounds of the present invention as inhibitors of plasma kallikrein can be determined using the relevant purified serine protease and appropriate synthetic substrates. The hydrolysis rate of the relevant serine protease on a chromogenic or fluorescent substrate was measured in the absence and presence of the compounds of the present invention. The determination was performed at room temperature or 37°C. The hydrolysis of the substrate resulted in the release of aminotrifluoromethylcoumarin (AFC), which was monitored by fluorescence spectroscopy by measuring the increase in emission at 510 nm under 405 nm excitation. A decrease in the rate of fluorescence change in the presence of the inhibitor indicates enzyme inhibition. Such methods are known to those skilled in the art. The results of this determination are expressed as the half-maximal inhibitory concentration (IC50). 50 or suppression constant K i .

[0387] Plasma kallikrein assays were performed in 50 mM HEPES buffer at pH 7.4 containing 150 mM NaCl, 5 mM CaCl2, and 0.1% PEG 8000 (polyethylene glycol; Fisher Scientific). The assays were performed using purified human plasma kallikrein (Enzyme Research Laboratories) at a final concentration of 0.5 nM and the synthetic substrate acetyl-KPR-AFC (Sigma#C6608) at a concentration of 100 mM.

[0388] Activity was determined by diluting the substrate stock solution at least tenfold to a final concentration ≤0.2 km in a solution containing the enzyme or an enzyme balanced with an inhibitor. The time required to achieve equilibrium between the enzyme and inhibitor was determined in a control experiment. The reaction was performed under linear progression conditions, and the increase in fluorescence was measured at 405 Ex / 510 Em nm. The values ​​were converted to the percentage of inhibition of the control reaction (after subtracting 100% inhibition). The IC50 was determined from the inflection point of the four-parameter logistic curve fitting. 50 Using Cheng Prusoff's equation Ki = IC 50 Calculate Ki using / (1+([S] / Km)).

[0389] The activity indicated by this assay suggests that the compounds of the present invention can be used therapeutically to treat or prevent various ophthalmic, cardiovascular, and / or cerebrovascular thromboembolic conditions in patients suffering from unstable angina, acute coronary syndrome, refractory angina, myocardial infarction, transient ischemic attack, atrial fibrillation, stroke such as thrombotic or embolic stroke, venous thrombosis, coronary and cerebral artery thrombosis, cerebral and pulmonary embolism, atherosclerosis, deep vein thrombosis, disseminated intravascular coagulation, re-occlusion or restenosis of recanalized vessels, hereditary angioedema, uveitis, posterior uveitis, wet age-related macular edema, diabetic macular edema, diabetic retinopathy, and retinal vein occlusion.

[0390] The plasma kallikrein IC50 of the selected compounds 50 (nM) and FXIa IC 50 (nM) is as follows:

[0391]

[0392]

[0393]

[0394]

[0395]

Claims

1. A compound of the following formula or a pharmaceutically acceptable salt thereof: Where X is N or CH; R 1 Selected from hydrogen, halogen, hydroxyl and C 1-6 alkyl; R 2 Selected from hydrogen, halogen, hydroxyl and C 1-6 alkyl; R 3 Selected from hydrogen, halogen, hydroxyl and C 1-6 Alkyl group, wherein the alkyl group is optionally substituted with 1 to 4 substituents, the substituents being independently selected from halogens, cyano groups, and OR groups. x ; R 4 Selected from hydrogen, halogen, hydroxyl and C 1-6 Alkyl; wherein the alkyl group is optionally substituted with 1-4 substituents, the substituents being independently selected from halogens, cyano groups, and OR. x ; R 5 It is NR 9 R 10 OR x ; Each R 6 Independently selected from hydrogen, halogen, hydroxyl and C 1-6 Alkyl group, wherein the alkyl group is optionally substituted with 1-3 halogens; Each R 7 Selected from hydrogen, halogen, hydroxyl and C 1-6 Alkyl group, wherein the alkyl group is optionally substituted with 1-3 halogens; or R 6 and R 7 They can form 3-6 membered cycloalkyl groups together with the carbon atoms to which they are attached, wherein the cycloalkyl groups are optionally substituted with one or two halogens; R 8 Selected from hydrogen; halogen; hydroxyl group; R x OR x ; Phenylacetylene; Indene; OR y It can be a monocyclic or bicyclic heteroaryl group; a heterocyclic group; and a C group that can be a monocyclic or bicyclic group. 3-6 Cycloalkyl; wherein the phenyl and heteroaryl groups are optionally substituted with 1-3 substituents, the substituents being independently selected from oxo, halogen, and R-type compounds. x OR x NR 9 R 10 NR 9 (C=O)R x NR 9 (C=O)OR x (C=O)OR x (C=O)NR 9 R y and OR y The cycloalkyl and heterocyclic groups are optionally substituted with 1-3 substituents, which are independently selected from oxo, halogen, and R groups. x and OR x ; R 9 Is it hydrogen or C? 1-3 alkyl; R 10 Is it hydrogen or C? 1-3 alkyl; R x Is it hydrogen or C? 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with 1-3 substituents selected from halogens and hydroxyl groups, R y It is phenyl, heterocyclic or C 3-6 Cycloalkyl groups, wherein the phenyl group is optionally substituted with 1-3 halogens, the heterocyclic group is optionally substituted with 1 or 2 oxygen groups, and the cycloalkyl group is optionally C-substituted. 1-6 Alkyl substitution; n is an integer between 0 and 2. The heterocyclic group mentioned therein refers to a stable non-aromatic monocyclic or bicyclic ring system having up to 10 atoms in each ring, and unless otherwise indicated, contains 1 to 4 heteroatoms selected from O, N, S, SO or SO2.

2. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R 1 It is halogen; R 2 It is halogen.

3. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R 3 It is hydrogen or methyl; R 4 It is either hydrogen or methyl.

4. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R 5 It is NH2.

5. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein n is 0 or 1.

6. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R 8 It is a phenyl group; wherein the phenyl group is optionally substituted with 1-3 substituents, the substituents being independently selected from halogens, R... x OR x NR 9 R 10 NR 9 (C=O)R x NR 9 (C=O)OR x (C=O)NR 9 (C=O)OR x R y and OR y .

7. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R 8 It is a phenyl group; wherein the phenyl group is optionally substituted with 1-3 substituents, the substituents being independently selected from halogens, R... x OR x R y and OR y .

8. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein n is 1; R 6 It is hydrogen; R 7 It is hydrogen.

9. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from any one of the following compounds 1-133:

10. A pharmaceutical composition comprising a compound according to any one of claims 1-9 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

11. Use of the composition according to claim 10 in the preparation of a medicament for treating visual impairment, diabetic retinopathy, diabetic macular edema, retinal vein occlusion, hereditary angioedema, diabetes, cerebral hemorrhage, nephropathy, cardiomyopathy, neuropathy, inflammation, septic shock, hypotension, cancer, adult respiratory distress syndrome, disseminated intravascular coagulation, blood clotting during cardiopulmonary bypass surgery, or postoperative bleeding in mammals.

12. Use of the composition according to claim 10 in the preparation of a medicament for the treatment of pancreatitis, inflammatory bowel disease, or arthritis in mammals.

13. Use of the composition according to claim 10 in the preparation of a medicament for the treatment of uveitis, wet age-related macular edema, diabetic macular edema, diabetic retinopathy, or retinal vein occlusion in mammals.

14. Use of the composition according to claim 10 in the preparation of a medicament for the treatment of posterior uveitis in mammals.

15. Use of the composition according to claim 10 in the preparation of a medicament for the treatment of diabetic retinopathy or diabetic macular edema in mammals.

16. Use of the composition according to claim 10 in the preparation of a medicament for the treatment of retinal vein occlusion in mammals.

17. Use of the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-9 in the preparation of a medicament for the treatment of uveitis, wet age-related macular edema, diabetic macular edema, diabetic retinopathy, or retinal vein occlusion in mammals in need of such treatment.

18. Use of the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-9 in the preparation of a medicament for the treatment of posterior uveitis in mammals in need of such treatment.

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