Heteroaromatic carboxamide derivatives as plasma kallikrein inhibitors

Abstract

Heteroaromatic carboxamides of formula (I), TIFF2023513906000091.tif37170(in the formula, R, R 1 , A 1 , A 2 , L 1 and L 2 (wherein R is as defined in the specification and claims) and pharmaceutically acceptable salts thereof can be used in methods of treating diseases that can be affected by inhibition of plasma kallikrein.

Description

Technical Field

[0001] The present invention relates to novel heteroaromatic carboxamide derivatives which are plasma kallikrein inhibitors and pharmaceutically acceptable salts thereof. Further, the present invention relates to pharmaceutical compositions and combinations containing said compounds, and their use in methods for treating diseases that can be affected by the inhibition of plasma kallikrein. In particular, the pharmaceutical compositions of the present invention are suitable for the prevention and / or treatment of diabetic complications, eye diseases and edema-related diseases, in particular, diabetic macular edema, age-related macular degeneration, choroidal neovascularization, hereditary angioedema, and cerebral edema after stroke.

Background Art

[0002] Plasma kallikrein (PKK) is a trypsin-like serine protease secreted by hepatocytes in the liver as an inactive plasma prekallikrein circulating in the plasma, either as a free zymogen or as a heterodimeric complex bound to high molecular weight kininogen, which upon activation results in active PKK capable of releasing kinins from kininogen in addition to the processing of other substrates. Kinins are powerful mediators of inflammation acting via G protein-coupled receptors such as bradykinin receptors.

[0003] PKK is thought to play a role in several inflammatory disorders, including hereditary angioedema (HAE), retinopathy or diabetic retinopathy, proliferative and non-proliferative retinopathy, diabetic macular edema (DME), clinically significant macular edema (CSME), cystoid macular edema (CME), CME after cataract extraction, CME induced by cryotherapy, CME induced by uveitis, endophthalmitis, CME after vascular occlusion (e.g., central retinal vein occlusion, branch retinal vein occlusion or hemi-retinal vein occlusion), retinal edema, complications associated with cataract surgery in diabetic retinopathy, hypertensive retinopathy, retinal trauma, atrophic and exudative age-related macular degeneration (AMD), polypoidal choroidal vasculopathy (PCV), choroidal neovascularization (CNV;For example, non-exudative choroidal neovascularization, posterior vitreous detachment (PVD), ischemia-reperfusion injury in all types of contexts, such as those related to tissue and / or organ transplantation, surgery-induced brain injury, focal cerebral ischemia, global cerebral ischemia, glioma-related edema, spinal cord injury, pain, ischemia, focal brain ischemia, neuropathy and cognitive impairment, deep vein thrombosis, stroke (including edema in the central nervous system after stroke), myocardial infarction, acquired angioedema, drug-related edema (including ACE inhibitor-induced edema and tissue plasminogen activator (tPA)-induced angioedema), high altitude cerebral edema, cytotoxic cerebral edema, osmotic cerebral edema, obstructive hydrocephalus, radiation-induced edema, lymphatic edema, traumatic brain injury, hemorrhagic stroke (e.g., stroke or subarachnoid hemorrhage), intracerebral hemorrhage, hemorrhagic transformation of ischemic stroke, extracranial trauma related to injury or surgery, cerebral aneurysm, arteriovenous malformation, reduction of blood loss during surgery (e.g., cardiac thoracic surgery such as cardiopulmonary bypass or coronary artery bypass grafting), psoriasis, disorders due to inflammatory components (such as multiple sclerosis), epilepsy, encephalitis, Alzheimer's disease, excessive daytime sleepiness, hypertension associated with essential hypertension, diabetes or hyperlipidemia, renal insufficiency, chronic kidney disease, heart failure, microalbuminuria, albuminuria, proteinuria, disorders related to increased vascular permeability (e.g., increased retinal vascular permeability, increased vascular permeability in the legs, feet, ankle joints), intracerebral hemorrhage, blood coagulation disorders such as thrombosis, deep vein thrombosis, coagulation resulting from fibrinolytic treatment, angina, angioedema, sepsis, arthritis (e.g., rheumatoid arthritis, osteoarthritis, infectious arthritis), lupus, gout, psoriasis, inflammatory bowel disease (IBD, e.g., ulcerative colitis (UC) and Crohn's disease (CD)), diabetes, diabetic complications, complications resulting from metabolic syndrome, infectious diseases, diseases related to astrocyte activation (such as Alzheimer's disease or multiple sclerosis), Parkinson's disease, amyotrophic lateral sclerosis, Creutzfeldt-Jakob disease, stroke, epilepsy and trauma (e.g., traumatic brain injury), allergic edema, e.g., airway obstruction in chronic allergic rhinosinusitis or perennial rhinitis; airway obstruction in acute asthma;It may have various associations with disorders such as serositis associated with systemic lupus erythematosus (SLE), acute respiratory distress syndrome (ARDS), coronavirus disease 2019 (COVID-19)-associated pneumonia, fibrotic diseases, liver fibrosis, non-alcoholic steatohepatitis (NASH), kidney injury and other diseases. PKK is also thought to play an important role in hypersensitivity reactions and thrombosis during hemodialysis.;

[0004] PKK inhibitors such as the compounds of the present invention are considered useful, for example, for the treatment of a wide range of disorders mentioned previously herein, and in particular, for the treatment of diabetic retinopathy and diabetic macular edema, or for the treatment of reducing retinal vascular permeability associated with edema-related diseases. PKK inhibitors should be particularly useful in the treatment of edema formation in diseases, for example, edema formation associated with edema-related diseases such as ischemia-reperfusion injury, retinopathy, or hereditary angioedema, macular edema and cerebral edema. PKK inhibitors are considered particularly useful for the treatment of retinopathy, for example, retinopathy associated with diabetes and / or hypertension, and for the treatment of macular edema, for example, macular edema associated with diabetes and / or hypertension. Other complications of diabetes such as cerebral hemorrhage, kidney injury, cardiomyopathy and neuropathy, all of which have an association with PKK, can also be considered as targets for PKK inhibitors.

[0005] PKK inhibitors suitable for therapeutic and / or prophylactic use should bind to PKK with high potency and high selectivity. Those inhibitors should be well absorbed from the gastrointestinal tract, metabolically stable enough, and have favorable pharmacokinetic properties. Those inhibitors should be non-toxic and show few side effects. Low molecular weight PKK inhibitors are known in the art. For example, these compounds are disclosed in WO2009 / 097141, WO2013 / 111107, WO2013 / 111108, WO2014 / 188211, WO2017 / 072020, WO2017 / 072021 and WO2018 / 192866. Summary of the Invention

[0006] In a first aspect, the present invention relates to a compound of formula (I) [Chemical formula] (wherein R is

[0007] [Chemical formula] selected from the group of R-G1 consisting of R 1 is selected from the group of R 1 -G1 consisting of H and F moiety =A 1 -CR=A 2 - is selected from the group of A-G1 consisting of =N-CR=N-, =N-CR=CH- and =CH-CR=N- moiety -L 1 =L 2 - is selected from the group of L-G1 consisting of -N=N-, -N=CH- and -CH=N-) It relates to isomers, tautomers, stereoisomers, metabolites, prodrugs, solvates, hydrates, co-crystals and salts thereof, in particular pharmaceutically acceptable co-crystals and salts thereof, or combinations thereof. In a second aspect, the present invention relates to a pharmaceutical composition which may together contain one or more compounds of formula (I) as defined hereinbefore or hereinafter in this specification, and / or their tautomers or pharmaceutically acceptable salts thereof, and one or more inert carriers and / or diluents.

[0008] In a third aspect, the present invention relates to a pharmaceutical composition which may together contain one or more compounds of formula (I) as defined hereinbefore or hereinafter in this specification, and / or their tautomers or pharmaceutically acceptable salts thereof, and one or more additional therapeutic agents, and one or more inert carriers and / or diluents. In a fourth aspect, the present invention relates to a compound of formula (I) as defined hereinbefore or hereinafter in the specification for use as a medicament, and / or a tautomer thereof or a pharmaceutically acceptable salt thereof. In a fifth aspect, the present invention is a method for treating, i.e., treating and / or preventing, a disease or condition that can be affected by inhibition of plasma kallikrein in a patient in need thereof, the method comprising administering to the patient one or more compounds of formula (I) as defined hereinbefore or hereinafter in the specification, and / or a tautomer thereof or a pharmaceutically acceptable salt thereof.

[0009] Furthermore, the present invention relates to the use of one or more compounds of formula (I) as defined hereinbefore or hereinafter in the specification, and / or a tautomer thereof or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating, i.e., treating and / or preventing, a disease or condition that can be affected by inhibition of plasma kallikrein. Furthermore, the present invention relates to a compound of formula (I) as defined hereinbefore or hereinafter in the specification for use in a method for treating, i.e., treating and / or preventing, a disease or condition that can be affected by inhibition of plasma kallikrein in a patient in need thereof, and / or a tautomer thereof or a pharmaceutically acceptable salt thereof. Further aspects of the present invention will be apparent to those skilled in the art from the foregoing and the following description and examples.

[0010] General terms and definitions Terms not specifically defined herein should be given the meaning that would be ascribed to them by those skilled in the art in light of the present disclosure and context. However, as used herein, unless stated to the contrary, the following terms have the indicated meanings and follow the following conventions. The terms "compound according to the invention", "compound of formula (I)", "compound of the invention", etc. mean the compounds of formula (I) according to the invention (their tautomers, stereoisomers and mixtures thereof, as well as their salts, in particular their pharmaceutically acceptable salts, as well as solvates, hydrates and co-crystals of such compounds, in particular their pharmaceutically acceptable co-crystals, solvates, hydrates and co-crystals of such tautomers, stereoisomers and their salts). Similarly, throughout this specification and the appended claims, unless specifically indicated otherwise, a given chemical formula or name includes tautomers, as well as all stereoisomers, optical isomers and geometric isomers (e.g., enantiomers, diastereomers, E / Z isomers, etc.), as well as their racemates, as well as mixtures of different proportions of the individual enantiomers, diastereomer mixtures, or any of the above forms in which such isomers and enantiomers are present, as well as salts (including pharmaceutically acceptable salts thereof), as well as their solvates (e.g., hydrates, etc., including solvates of the free compound or solvates of the salts of the compound), and their co-crystals (including pharmaceutically acceptable co-crystals thereof, and co-crystals of the free compound or its salts).

[0011] The phrase "pharmaceutically acceptable" as used herein refers to compounds, substances, compositions and / or dosage forms that are suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic reaction or other problems or complications, within the scope of sound medical judgment, and that exhibit a reasonable benefit / risk ratio. As used herein, "pharmaceutically acceptable salts" refers to derivatives of the disclosed compounds in which the parent compound is modified by making its acid or base salts. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids, etc. For example, such salts include salts derived from benzenesulfonic acid, benzoic acid, citric acid, ethanesulfonic acid, fumaric acid, gentisic acid, hydrobromic acid, hydrochloric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, 4-methyl-benzenesulfonic acid, phosphoric acid, salicylic acid, succinic acid, sulfuric acid, and tartaric acid. The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or free base forms of these compounds in water or in an organic diluent such as ether, EtOAc, EtOH, isopropanol or MeCN, or a mixture thereof, with a sufficient amount of an appropriate base or acid. For example, salts of acids other than those described above useful for purifying or isolating the compounds of the present invention (e.g., trifluoroacetate salts) also form part of the present invention.

[0012] As used herein, "pharmaceutically acceptable cocrystal" refers to a derivative of the disclosed compound, wherein the parent compound is modified by creating the cocrystal with the aid of one or more coformers. Similarly, solvates and / or cocrystals of salts of the disclosed compounds are also included. For example, coformers include hydrogen bond donors such as carboxylic acids, and hydrogen bond acceptors such as amines and amides. The pharmaceutically acceptable cocrystals of the present invention can be synthesized from the parent compound by methods known to those skilled in the art, including solid-based methods such as grinding in the solid state, melt extrusion molding, and melt crystallization, as well as liquid-based methods such as solution crystallization, solvent evaporation method, cooling crystallization, supercritical fluid-assisted crystallization, ultrasonic-assisted crystallization, spray drying, liquid-assisted grinding, and planetary ball milling. Where the compounds of the present invention are depicted in the form of a chemical name and, if any, as a formula with some differences, the formula shall prevail.

[0013] In the groups, radicals or moieties defined below, the number of carbon atoms is often specified before the group, e.g., C 1-6 -alkyl means an alkyl group or radical having 1 to 6 carbon atoms. An asterisk may be used in a subformula to indicate a bond attached to the defined core molecule. If there are more than one bond points, i.e., more than one asterisk, in the subformula, these asterisks may be further specified by the display within parentheses of the bonding part of the core molecule. The naming of the atoms of a substituent starts from the atom closest to the core or the group to which the substituent is attached. For example, the term "3-carboxypropyl group" represents the following substituent:

Chemical formula

Chemical formula

[0014] The term "C 1-n -alkyl" (n is an integer from 1 to n) means a saturated branched or linear acyclic hydrocarbon radical having 1 to n C atoms, either alone or in combination with another radical. For example, the term C 1-5 -alkyl is the radical H 3 C-, H 3 C-CH 2 -, H 3 C-CH 2 -CH 2 -, H 3 C-CH(CH 3 )-, H 3 C-CH 2 -CH 2 -CH 2 -, H 3 C-CH 2 -CH(CH3 )-, H 3 C-CH(CH 3 )-CH 2 -, H 3 C-C(CH 3 ) 2 -, H 3 C-CH 2 -CH 2 -CH 2 -CH 2 -, H 3 C-CH 2 -CH 2 -CH(CH 3 )-, H 3 C-CH 2 -CH(CH 3 )-CH 2 -, H 3 C-CH(CH 3 )-CH 2 -CH 2 -, H 3 C-CH 2 -C(CH 3 ) 2 -, H 3 C-C(CH 3 ) 2 -CH 2 -, H 3 C-CH(CH 3 )-CH(CH 3 )- and H 3 C-CH 2 -CH(CH 2 CH 3 )- are included. As used herein, the terms "treatment" and "treating" include both therapeutic, i.e., curative and / or palliative treatment, and prophylactic treatment, i.e., preventive treatment. Therapeutic treatment refers to the treatment of a patient who already has one or more of the foregoing conditions in an expressed, acute, or chronic form. Therapeutic treatment can be symptomatic treatment to relieve the symptoms of a particular condition, or etiological treatment to reverse or partially reverse the condition of the symptoms, or to stop or slow the progression of the disease.

[0015] Preventive treatment ("prevention") refers to treating a patient at risk of developing one or more of the above-mentioned conditions prior to the clinical onset of the disease in order to reduce the risk. The terms "treatment" and "treating" include administering one or more active compounds to prevent or delay the occurrence of symptoms or complications, and to prevent or delay the onset of a disease, condition or disorder, and / or to eliminate or control a disease, condition or disorder, and / or to reduce symptoms or complications associated with a disease, condition or disorder. When the present invention refers to a patient in need of treatment, the present invention is mainly concerned with treatment in mammals, particularly humans. The term "therapeutically effective amount" means an amount of a compound of the present invention that (i) treats or prevents a particular disease or condition, (ii) attenuates, ameliorates or eliminates one or more symptoms of a particular disease or condition, or (iii) prevents or delays the occurrence of one or more symptoms of a particular disease or condition described herein.

Best Mode for Carrying Out the Invention

[0016] The present invention discloses a novel heteroaromatic carboxamide derivative which is an effective plasma kallikrein (PKK) inhibitor, and has pharmacological and pharmacokinetic properties suitable for using the above carboxamide derivative as a medicament for treating diseases and / or conditions that can be affected by PKK inhibition, including but not limited to diabetic complications, eye diseases and edema-related diseases, particularly diabetic macular edema, age-related macular degeneration, choroidal neovascularization, hereditary angioedema and cerebral edema after stroke. The compounds of the present invention can achieve several advantages such as increased efficacy, high metabolic and / or chemical stability, high selectivity, safety and tolerance, increased solubility, increased permeability, desired plasma protein binding, improved bioavailability, improved pharmacokinetic profile, and the possibility of forming stable salts.

[0017] The compounds of the present invention In a first aspect of the present invention, a compound of formula (I)

Chemical formula

[0018] Accordingly, it is expected that the compound of formula (I) as defined hereinbefore or hereinafter in the present specification, or a pharmaceutically acceptable salt thereof, will be useful for the treatment of diseases and / or conditions that can be affected by PKK inhibition. Accordingly, according to one aspect of the present invention, a compound of formula (I)

Chemical formula

[0019] R: According to one embodiment, R is

Chemical formula

Chemical formula

[0020] According to another embodiment, R is

Chemical formula

Chemical formula

[0021] R 1 : According to one embodiment, R 1 is selected from the group R 1 -G1 consisting of H and F. According to another embodiment, R 1 consists of H 1- is selected from the group G2. According to another embodiment, R 1 is R consisting of F 1 - is selected from the group G3. A 1 , A 2 : According to one embodiment, moiety =A 1 -CR=A 2 - is selected from the group A-G1 consisting of =N-CR=N-, =N-CR=CH- and =CH-CR=N-. According to another embodiment, moiety =A 1 -CR=A 2 - is selected from the group A-G2 consisting of =N-CR=N-. According to another embodiment, moiety =A 1 -CR=A 2 - is selected from the group A-G3 consisting of =N-CR=CH-. According to another embodiment, moiety =A 1 -CR=A 2 - is selected from the group A-G4 consisting of =CH-CR=N-. Regarding the above definition of moiety =A 1 -CR=A 2 -, the order of the three subunits represents the arrangement of that moiety, i.e., for example, in the group A-G3, A 1 is N, and A 2 is CH.

[0022] L 1 , L 2 : According to one embodiment, -L 1 =L 2 - is selected from the group L-G1 consisting of -N=N-, -N=CH- and -CH=N-. According to another embodiment, -L 1 =L 2 - is selected from the group L-G2 consisting of -N=N-. According to another embodiment, -L 1 =L 2 - is selected from the group L-G3 consisting of -N=CH-. According to another embodiment, -L1 =L 2 - is selected from the group L-G4 consisting of -CH=N-. Sub-L 1 =L 2 Regarding the above definition of -, the order of the two subunits represents the arrangement of that part, i.e., for example, in the L-G3 group, L 1 is N, and L 2 is CH.

[0023] Lower embodiments of the more preferred compounds of formula (I) are described as embodiments (I-a) to (I-u) in Table 1 below, where the above definitions of the substituents are used. For example, the entry -G1 in column R and row (I-a) means that in embodiment (I-a), the substituent R is selected from the definition represented by R-G1. The same applies to the other variable groups incorporated in the general formula.

[0024]

Table 1

[0025] Particularly preferred compounds (including their tautomers, their salts, or any solvates, hydrates thereof) are

Chemical formula

[0026] Preparation The compounds according to the invention and their intermediates can be prepared using synthetic methods known to those skilled in the art and, for example, "Comprehensive Organic Transformations", 2 nd Edition, Richard C. Larock, John Wiley & Sons, 2010, and "March's Advanced Organic Chemistry", 7 thIt can be obtained using the synthetic methods described in the literature of organic synthesis that use the methods described in Edition, Michael B. Smith, John Wiley & Sons, 2013. Preferably, this compound is obtained in the same manner as the preparation method described in the experimental section and more fully described hereinafter in this specification. In some cases, the order adopted when implementing the reaction scheme can vary. These reaction variations, although known to those skilled in the art, are not described in detail herein and may also be used. The general method for preparing the compounds according to the present invention will be apparent to those skilled in the art upon considering the following scheme. The starting compounds are commercially available or can be prepared by the methods described in the literature or in this specification, or by similar or analogous methods. Prior to carrying out the reaction, the corresponding functional groups in the starting compounds may be protected using conventional protecting groups. These protecting groups may be cleaved again at suitable stages within the reaction sequence using methods well-known to those skilled in the art, as well as the methods described in the literature in, for example, "Protecting Groups", 3 rd Edition, Philip J. Kocienski, Thieme, 2005 and "Protective Groups in Organic Synthesis", 4th Edition, Peter G. M. Wuts, Theodora W. Greene, John Wiley & Sons, 2006.

[0027] Scheme 1:

Chemical formula

[0028] Scheme 1: The compound of formula (I’) can be prepared by reacting, in a suitable solvent (e.g., DCM, THF, 1,4-dioxane, DMF, N,N-dimethylacetamide, and 1-methyl-2-pyrrolidinone), a suitable acid of formula (II) (either a free acid or a carboxylate having a suitable metal cation such as Li + Na + K + etc.) with a suitable amine of formula (III) (either a free amine or a salt such as hydrochloride, hydrobromide, etc.) in the presence of a suitable coupling agent (e.g., O-(7-azabenzotriazol-1-yl)-N,N,N’,N’-tetramethyluronium hexafluorophosphate (HATU), O-(benzotriazol-1-yl)-N,N,N’,N’-tetramethyluronium tetrafluoroborate (TBTU), (benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyBOP), carbodiimide reagents, etc.) and a base (e.g., triethylamine, N,N-diisopropyl-ethylamine, pyridine, etc.) to form an amide bond. R, R 1 , A 1 , A 2 , L 1 and L 2has the meaning defined previously in this specification. Alternatively, the carboxylic acid is converted to a carboxylic acid chloride (e.g., using oxalyl chloride or thionyl chloride in DCM) and then directly coupled with the amine (III) in the presence of a suitable base (e.g., triethylamine, N,N - diisopropylethylamine, pyridine, etc.). When the amine (III) is used with a protecting group on the pyrazole ring (where PG is not H), this group is subsequently cleaved off by applying standard procedures reported in the organic chemistry literature. 2 - Trimethylsilylethyloxymethyl and tert - butyl esters are preferably cleaved under acidic conditions, e.g., using TFA or hydrochloric acid, in a solvent such as DCM, 1,4 - dioxane, isopropanol or EtOAc. 2 - Trimethylsilylethyloxymethyl can also be removed by using a fluoride source (e.g., n Bu 4 NF) in a suitable solvent such as THF. Benzyloxymethyl groups having an electron - donating group such as methoxy on the phenyl ring can also be cleaved under oxidative conditions (e.g., using cerium(IV) ammonium nitrate (CAN) or 2,3 - dichloro - 5,6 - dicyano - 1,4 - benzoquinone (DDQ)) or under acidic conditions (e.g., using TFA or hydrochloric acid).

[0029] Scheme 2: [Chemical formula] R 5 = C 1-4 - alkyl, benzyl

[0030] Scheme 2: R, R in Scheme 2 1 , A 1 , A 2 , L 1 and L 2 having the meaning defined previously in this specification, the acid of formula (II) where R 5Depending on the properties, it is preferably prepared from the corresponding ester (IV) by hydrolysis or hydrogenolysis. Esters of lower alkyl groups such as ethyl ester or methyl ester are preferably cleaved by hydrolysis with hydroxide salts such as NaOH, LiOH or KOH in a mixture of water and a suitable water-miscible solvent (e.g., THF, MeOH, EtOH, 1,4-dioxane or mixtures thereof) at ambient temperature or elevated temperature. The acid can be isolated either as a salt with a metal cation or as the free acid. tert-Butyl esters are preferably cleaved by treatment with an acid (e.g., hydrochloric acid or TFA) in a suitable solvent (e.g., DCM, 1,4-dioxane, MeOH, EtOH, THF, water or mixtures thereof). Benzyl esters are preferably cleaved by hydrogenolysis with a suitable catalyst (e.g., palladium on carbon) in a suitable solvent (e.g., EtOH, MeOH, THF, DCM or EtOAc) under a hydrogen atmosphere (preferably 1 - 5 bar).

[0031] Scheme 3

Chemical formula

[0032] Scheme 3: Some of the compounds (IV) can be prepared by the reaction of an alcohol (V) and an ester (VI) using the conditions of the Mitsunobu reaction (e.g., in a solvent such as THF, 1,4-dioxane, toluene, etc., combining triphenylphosphine or tri-n-butylphosphine with, for example, diethyl azodicarboxylate (DEAD), diisopropyl azodicarboxylate (DIAD) or di-tert-butyl azodicarboxylate (DBAD)). R, R 1 、A 1 、A 2 、L 1 and L 2has the meaning defined previously in this specification. Alcohol (V) can have a desired residue R on the heteroaromatic ring or, alternatively, a leaving group for later introduction of R. Alternatively, some of the compounds (IV) can be obtained by reacting alcohol (V) with ester (VI) in a suitable solvent (e.g., MeCN) at a high temperature (20 - 120 °C) in the presence of a Lewis acid or a Bronsted acid (e.g., 4-toluenesulfonic acid).

[0033] Scheme 4

Chemical formula

[0034] Scheme 4: Some of the compounds (IV) can also be prepared by reacting a compound (VII) having a leaving group such as Cl, Br or mesyloxy (methanesulfonyloxy) at the heteroarylmethyl position with ester (VI) in a suitable solvent (e.g., THF, DMF) in the presence of a suitable base (e.g., sodium hydride, cesium carbonate, potassium carbonate or triethylamine). R, R 1 , A 1 , A 2 , L 1 and L 2 have the meaning defined previously in this specification. Compound (VII) can have a desired residue R on the heteroaromatic ring or, alternatively, a substitutable group for later introduction of R.

[0035] Scheme 5

Chemical formula

[0036] Scheme 5: In Scheme 5, R, R 1 , A 1 and A 2 Some esters of formula (IV’) in which R, R

[0037] Scheme 6 [Chemical formula] R 2 = CHF 2 , CF 3 , or Cl, Br, I, HCCH 2 , CHO, CN, COOR 5 etc., groups that allow for the later introduction of CHF 2 and / or CF 3 R R 6 =COOR 5 , CHO, CH 2 OH, [Chemical formula] R 5 =C 1-4 -alkyl or benzyl; LG = F, Cl, Br, I, S(O) 0,1,2 and other leaving groups

[0038] Scheme 6: The intermediate of formula (XV) can be prepared from the aromatic compound (XIII) and the amine (XIV) either by a nucleophilic substitution reaction on the heteroaromatic ring or by a transition metal-catalyzed coupling reaction. A in Scheme 6 1 、A 2 and R have the meanings defined previously herein. The nucleophilic substitution of the leaving group on the heteroaromatic ring in (XIII) by N in the compound (XIV) can be carried out at ambient temperature or elevated temperature, in a suitable solvent (e.g., THF, 1,4-dioxane, DMF, DMSO), in the presence of a suitable base (e.g., sodium hydride, cesium carbonate, potassium carbonate, N,N-diisopropyl-ethylamine). The coupling reaction catalyzed by a transition metal is carried out using the chloride, bromide or iodide of the compound (XIII), in the presence of a base and in a suitable solvent, using a suitable copper salt or palladium salt, or their complexes (which may be combined with additional ligands), and is preferably carried out in the same manner as the procedures reported in the literature of organic chemistry, which are referred to as Ullmann coupling reactions or Buchwald / Hartwig coupling reactions.

[0039] Scheme 7

Chemical formula

[0040] Scheme 7: The intermediate of formula (XII) can be obtained from the compound (XIII’) by a transition metal-catalyzed coupling reaction with a suitable difluoromethyl or trifluoromethyl nucleophile or their precursors. R, R 1 、A 1 、A2 , L 1 and L 2 has the meaning defined previously in this specification. In a suitable solvent (e.g., NMP or DMF), at ambient temperature or elevated temperature, i.e., 20 - 150 °C, the compound of formula (XII) can be obtained with difluoromethyltrimethylsilane or trifluoromethyltrimethylsilane as individual nucleophile precursors in combination with a copper salt (e.g., CuI) and a base (e.g., CsF or KF). F 3 Using a preformed nucleophile that already incorporates a catalyst such as CCu(phen) (phen = phenanthroline), compound (XII) can be obtained similarly under the reported conditions (e.g., in DMF, 80 - 100 °C; see, for example, Org. Lett. 2014, 16, 1744 - 1747).

[0041] Compound (XI) can also be obtained from halide (XIII’) by a three - step synthetic sequence. Thus, halide (XIII’) is vinylated using a suitable vinyl nucleophile, e.g., vinyl zinc chloride or vinylboronic acid or vinylboronic acid ester, and the reported conditions for the so - called Negishi coupling reaction (e.g., in THF, 30 - 60 °C, using Pd(PPh 3 ) 4 or the Suzuki coupling reaction (e.g., in 1,4 - dioxane or DMF, 60 - 110 °C, using Na 2 CO 3 and in the presence of water, using PdCl 2 (dppf)) to form compound (IX). Next, compound (IX) is subjected to C = C cleavage conditions (e.g., ozonolysis in DCM at - 70 °C followed by reductive quenching with PPh 3 or Me 2 S, or dihydroxylation with OsO 4 in acetone, tert - butanol and water at ambient temperature followed by NaIO 4is subjected to glycol cleavage). Subsequently, in a suitable solvent (e.g., DCM or toluene), from ambient temperature to high temperature, in the presence of an optionally catalytic amount of methanol or fluoride salt, using a suitable deoxofluorinating agent (e.g., DAST (Et 2 NSF 3 ), Deoxofluor ((MeOCH 2 CH 2 )) 2 NSF 3 ), or XtalFluor-E ([Et 2 NSF 2 BF 4 )) to deoxofluorinate the aldehyde (X), this route is completed.

[0042] Scheme 8

Chemical Structure

[0043] Scheme 8: The pure amines as enantiomers of formula (III.1) and (III’.1) can be prepared from ketones (XVI) and (XVI’), respectively, as illustrated in Scheme 8. The ketone of formula (XVI) is enantioselectively reduced under various conditions reported in the literature of organic chemistry (for example, J. Am. Chem. Soc. 1995, 117, 7562-3; Org. Lett. 2010, 12, 1756-9; Org. Proc. Res. Dev. 2006, 10, 949-958; Tetrahedron: Asymmetry 2003, 14, 2659-2681; Tetrahedron Lett. 2014, 55, 3635-40; and the references cited therein) to give a pure or enantiomerically enriched alcohol (not shown) as an enantiomer of formula (XVII) or (XVII’), which is derived from the ketone (XVI’). Next, this alcohol is used in a Mitsunobu reaction or a Mitsunobu-type reaction (in a suitable solvent (for example, THF, 1,4-dioxane, EtOAc, benzene, toluene, etc.), for example, triphenylphosphine or tri-n-butylphosphine, combined with dimethyl azodicarboxylate or diethyl azodicarboxylate or diisopropyl azodicarboxylate, di-(4-chlorobenzyl) azodicarboxylate, dibenzyl diazodicarboxylate, DBAD, bis-(dimethylamide) azodicarboxylate, dipiperidyl azodicarboxylate or dimorpholide azodicarboxylate) with phthalimide or (tert-Bu-OCO) 2Reacting with sufficiently acidic N-H containing molecules such as NH can result in the introduction of an N-residue with an inverted configuration at the stereocenter (→(XVIII)). Alternatively, phosphoryl azide (e.g., diphenylphosphoryl azide) can be used to replace the OH in (XVII) with an azide under inversion of the configuration of the adjacent carbon atom. The amino group can be liberated from the phthalimide group to obtain the compound of formula (III.1) by treating, for example, with hydrazine, hydroxylamine, methylamine, n-butylamine or ethanolamine in a suitable solvent (e.g., EtOH, MeOH, MeCN, THF, dioxane, DMSO, N,N-dimethylacetamide, water or a mixture thereof), with heating if necessary. tert-Bu-O-CO is preferably removed under acidic conditions (e.g., using TFA or hydrochloric acid) to obtain the amine (III.1). The azide can be reduced to the amine (III.1) with, for example, hydrogen in the presence of a transition metal (e.g., Pd on carbon, Raney-Ni, PtO 2 etc.) or a phosphine (e.g., triphenylphosphine).

[0044] Alternatively, the compound (III.1) can be treated with a titanium alkoxide (e.g., Ti(OEt) 4 or Ti(O i Pr) 4) In the presence of, a three-step synthetic sequence using pure tert-butanesulfinamide as an enantiomer is obtained from ketone (XVI), and a corresponding tert-butylsulfinylated imine that is pure as an enantiomer is generated. This imine can be diastereoselectively reduced to the corresponding tert-butylsulfinylated amine using a hydride (e.g., lithium borohydride or sodium borohydride, L-selectride, diisobutylaluminum hydride, etc.) in a suitable solvent (e.g., THF, toluene, MeOH, etc., depending on the hydride source used). The tert-butylsulfinyl group can be cleaved and removed using an acid (e.g., TFA or hydrochloric acid) in a suitable solvent (e.g., toluene, DCM, dioxane, alcohol, water, etc.) at ambient temperature or elevated temperature.

[0045] Scheme 9

Chemical formula

[0046] Scheme 9: Compound (XVI) can be obtained from the reported ester (XIX) (or the corresponding higher alkyl ester, e.g., ethyl, propyl, isopropyl or tert-butyl ester) in an order consisting of five or six reaction steps. Compound (XIX) can be derivatized on one of its N atoms with a wide range of protecting groups reported in the organic chemistry literature. For example, compound (XIX) can be treated with a base (e.g., hydrides such as sodium hydride, hydroxides such as sodium hydroxide, carbonates such as sodium carbonate or potassium carbonate, alcoholates such as lithium methoxide or potassium tert-butyrate, organic amines such as triethylamine, Hunig's base, DABCO, DBN or DBU, phosphazenes such as P 2 Et phosphazene, amides such as lithium diisopropylamide or lithium hexamethyldisilazide) in a suitable solvent (e.g., benzene, toluene, DCM, THF, dioxane, EtOAc, ACN, DMF, N,N-dimethylacetamide, N-methylpyrrolidinone, etc., depending on the nature of the base used) and converted to compound (XX) by simultaneous or subsequent reaction with an electrophile (chloride, bromide, iodine, alkylsulfonyloxy or arylsulfonyloxy, leaving groups such as alkyloxy, acyloxy, etc.) of a suitable protecting group (e.g., 2-trimethylsilylethyloxymethyl chloride for introducing 2-trimethylsilylethyloxymethyl as a protecting group). Compound (XX) can be reacted with the corresponding halogen (e.g., N-chlorosuccinimide for Cl, N-bromosuccinimide for Br or Br 2 ; for I, N-iodosuccinimide, I 2Or ICl, and may be carried out in the presence of additives such as silver salts or acids) using a suitable electrophilic source, and can be chlorinated, brominated or iodinated. For example, iodine can be introduced using N-iodosuccinimide and TFA in MeCN to give compound (XXI). Next, compound (XXII) can be prepared from the corresponding halide (e.g., iodide (XXI)) using a one-step or two-step synthetic route including a Heck coupling reaction with either acrolein dialkyl acetal (e.g., acrolein dimethyl acetal) or acrylic acid ester (e.g., methyl acrylate) (broadly included in the literature of organic chemistry, e.g., Catalysts 2017, 7, 267, and the references cited therein). The use of the latter acrylic acid ester as the coupling partner requires an additional step of reducing the formed olefin bond, which can be carried out using hydrogen in the presence of a transition metal catalyst (e.g., Pd such as palladium on carbon, Ni such as Raney-Ni, Pt such as platinum oxide, Rh such as rhodium on carbon, etc.) in a suitable solvent (e.g., alcohols such as DCM, dioxane, THF, EtOAc, MeOH, water, etc.). The ketoester (XXIII) can be prepared from a low temperature to a high temperature (-78 °C to 100 °C depending on the base and solvent used) in a suitable solvent (e.g., benzene, toluene, dioxane, THF, alcohol, etc. depending on the base used) in the presence of a base (e.g., hydrides such as sodium hydride, alcoholates such as lithium methoxide or potassium tert-butyrate, organic amines such as DBU, P 2Compound (XXII) can be produced by treating it with a phosphazene such as Et phosphazene, an amide such as lithium diisopropylamide, lithium hexamethyldisilazide, sodium hexamethyldisilazide or potassium hexamethyldisilazide. Hydrolysis of the ester group in compound (XXIII) and subsequent decarboxylation can be achieved by stirring this compound at 0 to 140 °C in a solvent (such as dioxane, THF, MeCN, DMF, N,N-dimethylacetamide, DMSO, alcohol, water, etc., or a mixture thereof) (in the presence of a base (such as sodium hydroxide), a halogenated salt such as lithium iodide or lithium chloride, or an acid (such as hydrochloric acid)), to obtain ketone (XVI). The overall procedure can be similarly applied to the isomerically protected compound (XX’) to obtain ketone (XVI’), and does not necessarily depend on the use of a protecting group, and thus can be carried out without a protecting group (PG = H).

[0047] As described above, the compound of formula (I) can be converted into salts, especially pharmaceutically acceptable salts for pharmaceutical use. As used herein, "pharmaceutically acceptable salts" refers to derivatives of the disclosed compounds in which the parent compound is modified by making its acid or base salts. The compounds according to the invention can advantageously also be obtained using the methods described in the following examples, which can also be combined for this purpose using methods known to those skilled in the art from the literature. Accordingly, according to another aspect of the invention, a method for synthesizing the compound of formula (I) is provided. According to another aspect of the invention, a synthetic intermediate of the compound of formula (I) is provided.

[0048] Pharmacological activity The activity of the compounds of the invention can be demonstrated using the following assays. Biological methods The ability of the compound of formula (I) to inhibit plasma kallikrein (PKK), factor XIIa (FXIIa), factor XIa (FXIa), factor Xa (FXa), factor IIa (alpha-thrombin; FIIa), plasmin, trypsin, tissue kallikrein 1 (TK1), factor VIIa (FVIIa), or factor VIIa, tissue factor, phospholipid and CaCl 2 complexed with FVIIa (FVIIa / TF / PL / CaCl 2 ) is determined using the following biochemical assay in assay buffer (100 mM Tris, 150 mM NaCl, adjusted to pH 7.8 with HCl and containing 0.1% (w / v) BSA and 0.05% (v / v) Tween 20) in the presence of 1% (v / v) DMSO.

[0049] Inhibition assessment of PKK using endpoint assay Human PKK (0.01 U / mL; Enzyme Research Laboratories) or rat PKK (0.625 nM; generated in-house) is incubated at room temperature for 1 hour in assay buffer with 0.10 μM fluorescent substrate H-Pro-Phe-Arg-AMC (I1295, from Bachem) and various concentrations of the test compound. Subsequently, PPACK II (Calbiochem) is added as a stop solution to a final concentration of 1 μM, and fluorescence is measured using an Envision Reader (PerkinElmer) with an excitation wavelength setting of 355 nm and an emission wavelength setting of 460 nm.

[0050] The IC 50 values of the compounds according to the present invention are shown in the following table. The compound numbers correspond to the example numbers in the experimental section.

Table 2

[0051] Inhibition assessment of PKK in kaolin-activated human PPP Platelet-poor plasma (PPP) obtained from human whole blood anticoagulated with sodium citrate (Na-Citrat) was incubated with various concentrations of test compounds together with kaolin at 25, 75, 250 or 750 μg / mL in an assay buffer at 37 °C for 20 minutes, thus obtaining the concentration response for each kaolin dose used against the test compound. Thereafter, 0.25 mM of the fluorescent substrate H-Pro-Phe-Arg-AMC (I1295, manufactured by Bachem) was added to the above mixture, and measurements were taken at excitation and emission wavelengths of 350 nm and 450 nm, respectively, at kinetic intervals of 2 minutes for 12 minutes using a Spectramax M5 (Molecular Devices). pIC50 and pIC90 values were obtained from a 4x / y-plot (x = logM, compound; y = delta rfu / min) fitted to GraphPad prism 7.0 (Equation: log(agonist) vs. response, i.e., Find ECanything; the four concentration-response curves obtained for the test compound (each obtained using a different kaolin dose) were fitted using a global fitting procedure to obtain a common pIC50 or pIC90 value).

[0052] IC of the compound according to the present invention 90 values are shown in the following table. The compound numbers correspond to the example numbers in the experimental section. [Table 3]

[0053] Inhibition of PKK (K i ) evaluation Human PKK (1.78 nM or 0.025 U / mL; Enzyme Research Laboratories) is incubated at 24 °C in assay buffer with 0.25 mM of the fluorescent substrate H-Pro-Phe-Arg-AMC (I1295, Bachem) and various concentrations of the test compound. Measurements are taken using a Spectramax M5 (Molecular Devices) at an excitation wavelength of 350 nm and an emission wavelength of 450 nm, at kinetic intervals of 2 minutes for 16 minutes. Inhibition of FXIIa (K i ) evaluation Human FXIIa (47.5 nM or 1.1 U / mL; Enzyme Research Laboratories) is incubated at 24 °C in assay buffer with 0.5 mM of the chromogenic substrate S2302 (Chromogenix) and various concentrations of the test compound. Measurements are taken using a Spectramax M5 (Molecular Devices) at kinetic intervals of 2 minutes for 16 minutes, measuring the optical absorbance at 405 nm.

[0054] Inhibition of FXIa (K i ) evaluation Human FXIa (0.5 nM or 0.016 U / mL; Enzyme Research Laboratories) is incubated at 24 °C in assay buffer with 0.25 mM of the fluorescent substrate Boc-Glu(OBzl)-Ala-Arg-AMC·HCl (I1575, Bachem) and various concentrations of the test compound. Measurements are taken using a Spectramax M5 (Molecular Devices) at an excitation wavelength of 350 nm and an emission wavelength of 450 nm, at kinetic intervals of 2 minutes for 16 minutes. Inhibition of FXa (K i ) evaluation Human FXa (0.86 nM or 0.01 U / mL; Enzyme Research Laboratories) is incubated at 24 °C in assay buffer with 0.5 mM chromogenic substrate S2765 (Chromogenix) and various concentrations of test compound. Measurements are taken using a Spectramax M5 (Molecular Devices) at kinetic intervals of 2 minutes for 16 minutes, measuring the optical absorbance at 405 nm.

[0055] Inhibition of FIIa (K i ) evaluation Human FIIa (44.6 nM or 5 U / mL; Enzyme Research Laboratories) is incubated at 24 °C in assay buffer with 0.5 mM chromogenic substrate S2238 (Chromogenix) and various concentrations of test compound. Measurements are taken using a Spectramax M5 (Molecular Devices) at kinetic intervals of 2 minutes for 16 minutes, measuring the optical absorbance at 405 nm. Inhibition of plasmin (K i ) evaluation Human plasmin (64.1 nM or 0.0275 U / mL; Enzyme Research Laboratories) is incubated at 24 °C in assay buffer with 0.3 mM chromogenic substrate S2251 (Chromogenix) and various concentrations of test compound. Measurements are taken using a Spectramax M5 (Molecular Devices) at kinetic intervals of 2 minutes for 16 minutes, measuring the optical absorbance at 405 nm.

[0056] Inhibition of trypsin (K i ) evaluation Human trypsin (4.54 nM or 250 U / mL; Calbiochem) is incubated at 24 °C with 0.5 mM of the chromogenic substrate S2222 (Chromogenix) and various concentrations of test compounds in the assay buffer. Measurements are taken using a Spectramax M5 (Molecular Devices) at kinetic intervals of 2 minutes for 16 minutes to measure the optical absorbance at 405 nm. Inhibition of TK1 (K i ) evaluation Prior to the assay, human TK1 (R&D Systems) is activated by incubation at 37 °C for 15 minutes with human trypsin (Calbiochem) at a ratio of 1:10,000. To assay for TK1 inhibitory activity, the activated TK1 (31.25 nM or 1 U / mL) is incubated at 24 °C with 0.1 mM of the fluorescent substrate H-Pro-Phe-Arg-AMC (I1295, manufactured by Bachem) and various concentrations of test compounds in the assay buffer. Measurements are taken using a Spectramax M5 (Molecular Devices) at kinetic intervals of 2 minutes for 16 minutes with the above settings of an excitation wavelength of 350 nm and an emission wavelength of 450 nm. The K i values of the compounds according to the present invention are shown in the following table. The compound numbers correspond to the example numbers in the experimental section. [Table 4]

[0057] Inhibition of FVIIa (K i ) evaluation Human FVIIa (0.86 nM or 0.01 U / mL; Enzyme Research Laboratories) is incubated at 24 °C in assay buffer with 1.5 mM chromogenic Pefachrome® FVIIa (Loxo) and various concentrations of test compound. Measurements are taken using a Spectramax M5 (Molecular Devices) at kinetic intervals of 2 minutes for 16 minutes, measuring the optical absorbance at 405 nm. FVIIa / TF / PL / CaCl 2 Inhibition of (K i ) evaluation Human FVIIa (300 nM or 585 U / mL; Enzyme Research Laboratories) is incubated at 24 °C in assay buffer with 1.5 mM chromogenic Pefachrome® FVIIa (Loxo), various concentrations of test compound, 10 mM CaCl 2 * 2H 2 O and 13.3% (v / v) Dade® Innovin® (Siemens; OQUMI94E0002(5534), which contains recombinant human tissue factor synthetic phospholipid (thromboplastin)). Measurements are taken using a Spectramax M5 (Molecular Devices) at kinetic intervals of 2 minutes for 16 minutes, measuring the optical absorbance at 405 nm.

[0058] pIC 50 and pK i value calculation For the time interval of 2 - 12 minutes after the start of the assay, the average V max values (expressed either as delta OD / min for assays using chromogenic substrates or delta RFU / min for assays using fluorogenic substrates) are plotted against the log of the concentration in molar concentration of the inhibitor compound evaluated. Next, pIC 50Values were fitted using a four - parametric fitting procedure with GraphPad Prism (version 6; GraphPad Software, Inc.). The K of the substrate used was obtained by correcting the IC values for each of the following equations: M values for each of the K 50 values. (See Table A for the obtained K values of the substrate used): i M M

Number

[0059]

Table 5

[0060] Evaluation of permeability Caco - 2 cells (1 - 2x10 5 cells / cm 2 area) were seeded on filter inserts (Costar transwell polycarbonate or PET filters, 0.4 μm pore size) and cultured (DMEM) for 10 - 25 days. The compound was dissolved in an appropriate solvent (a 1 - 20 mM stock solution such as DMSO). The stock solution contained HTP - 4 buffer (128.13 mM NaCl, 5.36 mM KCl, 1 mM MgSO 4 4, 1.8 mM CaCl 2 4, 4.17 mM NaHCO 3 4, 1.19 mM Na 2 HPO 4 x7H 2 2O, 0.41 mM NaH 2 PO 4 xH 2Dilute with O, 15 mM HEPES, 20 mM glucose, pH 7.2) to prepare a transport solution (0.1 - 300 μM compound, final DMSO <= 0.5%). The transport solution (TL) is applied to the apical donor or basolateral donor (2 filter replicates each) to measure the permeability of A - B or B - A respectively. The receiver side contains HTP - 4 buffer supplemented with 0.25% BSA. Samples are taken from the donor and also from the receiver side at the start and end of the experiment and at various times up to a maximum of 2 hours for concentration measurement by HPLC - MS / MS or scintillation counting. The sampled volume of the receiver is exchanged with fresh receiver solution.

[0061] Evaluation of metabolic stability in human or rat liver microsomes Assay the metabolic degradation of the test compound at 37 °C with pooled human liver microsomes (HLM) or rat liver microsomes (RLM). The final incubation volume, which is 60 μl per time point, contains TRIS buffer (0.1 M) at pH 7.6, magnesium chloride (5 mM), microsomal protein (HLM: 1 mg / mL, RLM: 0.5 mg / mL) and the test compound at a final concentration of 1 μM at room temperature. After a short pre - incubation period at 37 °C, start this reaction by adding reduced beta - nicotinamide adenine dinucleotide phosphate (NADPH, 1 mM), and terminate it at various time points by transferring an aliquot to the solvent. Furthermore, monitor the NADPH - independent degradation in an incubation without NADPH and terminate it at the last time point. The post - quenched incubation is pelleted by centrifugation (10000 g, 5 minutes). For the amount of the parent compound, assay an aliquot of the supernatant by LC - MS / MS. The half - life (t1 / 2 INVITRO) is determined by the slope of the semi - logarithmic plot of the concentration - time profile.

[0062] Evaluation of metabolic stability in human or rat hepatocytes The metabolic decomposition of the test compound is assayed in a hepatocyte suspension. Human or rat hepatocytes are recovered from cryopreservation and then incubated in Dulbecco's modified Eagle's medium supplemented with 3.5 μg of glucagon / 500 ml, 2.5 mg of insulin / 500 ml, and 3.75 mg / 500 ml of hydrocortisone, containing 5% or 50% human or rat serum, or in the absence of serum. After a 30-minute pre-incubation (37 °C, 10% CO 2 ) in a cell culture incubator, the test compound solution is spiked into the hepatocyte suspension to obtain a final cell density of 1.0 * 106 cells / ml, a final test compound concentration of 1 μM, and a final DMSO concentration of 0.05%. The cells are incubated for 6 hours (in an incubator, horizontal shaker), and samples are removed from the incubation at 0, 0.5, 1, 2, 4, and 6 hours. The samples are quenched with acetonitrile and pelleted by centrifugation. The supernatant is transferred to a 96-deep well plate and prepared for analysis of the decrease in the parent compound by HPLC-MS / MS.

[0063] CL int is calculated as follows: CL int = dose / AUC = (C0 / CD) / (AUD + clast / k) × 1000 / 60 C0: initial concentration in the incubation [μM], CD: cell density of viable cells [10e6 cells / mL], AUD: area under the data curve [μM x h], clast: concentration at the last data point [μM], k: slope of the regression line for the decrease of the parent [h -1 . The calculated in vitro hepatic intrinsic clearance can be scaled up to in vivo intrinsic hepatic clearance and used to predict in vivo hepatic blood clearance (CL) by using a liver model (well-stirred model).

[0064] Evaluation of Plasma Protein Binding Using this equilibrium dialysis (ED) technique, an approximate in vitro binding rate of the test compound to plasma proteins is determined by applying a Dianorm Teflon dialysis cell (Micro 0.2). Each dialysis cell consists of a donor chamber and an acceptor chamber, separated by an ultra-thin semi-permeable membrane with a molecular weight cut-off of 5 kDa. The stock solution of each test compound is prepared in 1 mM DMSO and serially diluted to obtain a final test concentration of 1 μM. The subsequent dialysis solution is prepared in plasma (supplemented with NaEDTA as an anticoagulant), and a fixed volume of the dialysis solution of the test compound, which is 200 μL in plasma, is dispensed into the donor (plasma) chamber. A fixed volume of 200 μL of dialysis buffer (100 mM potassium phosphate, pH 7.4) is dispensed into the buffer (acceptor) chamber. Incubation is carried out at 37 °C under rotation for 2 hours to establish equilibrium. At the end of the dialysis period, fixed volumes obtained from the donor chamber and the acceptor chamber respectively are transferred to reaction tubes, spiked with an internal standard solution, and processed for HPLC-MS / MS analysis. The concentration of the analyte in the fixed volume of the sample is quantified by HPLC-MS / MS against an external calibration curve. The binding rate is calculated using the following formula: % Binding = ((Concentration in plasma - Concentration in buffer) / Concentration in plasma) X 100

[0065] Evaluation of solubility The solubility of the test compound in water is determined by comparing the amount dissolved in buffer with the amount in an ACN / water (1 / 1) solution. Starting from a 10 mM DMSO stock solution, fixed volumes are diluted with acetonitrile / water (1 / 1) or buffer respectively. After shaking for 24 hours, this solution is filtered and analyzed by LC-UV. The amount dissolved in buffer is compared with the amount in the ACN solution. The solubility will typically be measured at a DMSO concentration of 2.5% and range from 0.001 to 0.125 mg / mL. If more than 90% of the compound dissolves in buffer, this value is noted as ">". Evaluation of pharmacokinetic characteristics in rodents The test compound is administered either intravenously to fed rats or orally to fasted rats. Blood samples are collected at several time points after administration of the test compound, anticoagulated, and centrifuged. The concentration of the analyte, i.e., the administered compound and / or metabolite, is quantified in plasma samples. PK parameters are calculated using the non-compartmental method. AUC and Cmax are normalized to a dose of 1 μmol / kg.

[0066] Evaluation of the inhibition of reactions catalyzed by cytochrome P450 isoenzymes The inhibition of reactions catalyzed by cytochrome P450 isoenzymes by the test compound is assayed at 37 °C using human liver microsomes. All assays are performed in a 384-well plate using a robotic system. The test compound is spotted directly from a DMSO stock solution onto the incubation plate by acoustic liquid dispensing (using the Labyte ECHO® system). The final incubation volume contains, in duplicate, TRIS buffer (0.1 M), MgCl 2 (5 mM), human liver microsomes, a specific cytochrome P450 isoenzyme-substrate, and the test compound at five different concentrations or the compound is not included (e.g., at a maximum concentration of 50 μM and then serially diluted 1:4). After a short pre-incubation period, the reaction is initiated with the cofactor (NADPH, 1 mM), and the incubation is cooled to 8 °C and subsequently stopped by adding one volume of acetonitrile. An internal standard solution, usually a stable isotope of the metabolite formed, is added after quenching of the incubation. The peak areas of the analyte (= metabolite formed) and the internal standard are determined by LC-MS / MS. The ratio of the obtained peak area of the analyte to the internal standard in these incubations is compared to the control activity without the test compound. Within the scope of each assay, the IC 50 of the positive control inhibitor is determined. The experimental value of IC50 is calculated by least-squares regression according to the following formula:

[0067] % Control Activity = (100% of control activity / (1 + (I / IC 50 ))S)) - b (where I = inhibitor concentration S = slope factor B = background activity) If the inhibition of the reaction is already > 50% at the lowest concentration of the test compound, IC 50 is assigned as "< lowest concentration tested" (usually < 0.2 μM). If the inhibition of the reaction is still < 50% at the highest concentration of the test compound, IC50 is assigned as "> highest concentration tested" (usually > 50 μM).

[0068] Evaluation of mechanism-based inhibition (MBI) of midazolam turnover catalyzed by cytochrome P450 3A4 Mechanism-based inhibition against CYP3A4 is assayed in human liver microsomes (0.02 mg / ml) containing midazolam (15 μM) as a substrate. The test compound is pre-incubated at 37 °C for 0, 10 or 30 minutes at concentrations of 5 μM and 25 μM in the presence of NADPH with human liver microsomes (0.2 mg / ml). After pre-incubation, the incubate is diluted 1:10 and the substrate midazolam is added for the main incubation (15 minutes). The main incubate is quenched with acetonitrile and the formation of hydroxy-midazolam is quantified by LC / MS-MS. The turnover rate in pmol / min per mg of protein is calculated and the activity after 10 and 30 minutes of pre-incubation time is compared to the activity of the 0-minute pre-incubation of the individual compound / concentration (%CTRL = % of control at 0 minutes of the individual compound / concentration). Further, to clarify the competitive inhibition effect, the turnover rate is expressed relative to the turnover rate of the substrate reaction without the compound (%TR = % turnover rate without the compound).

[0069] Treatment method In another aspect of the invention, the compound of formula (I) or a pharmaceutically acceptable salt thereof has properties suitable for use in therapy and / or prophylaxis, i.e., for use as a medicament. In particular, the compound of formula (I) or a pharmaceutically acceptable salt thereof, and pharmaceutical compositions containing them, can affect in a patient by inhibition of plasma kallikrein, for example, mediated by unwanted PKK activity, or in which inhibition of PKK is beneficial, i.e., can be useful for the treatment, i.e., therapy and / or prophylaxis (prevention), of diseases or conditions.

[0070] Diseases and conditions that can be affected by inhibition of PKK, for example, mediated by unwanted PKK activity, or in which inhibition of PKK is beneficial, are, for example, those mentioned in the background of the invention section, in particular, diabetic complications, diabetic retinopathy, proliferative and non - proliferative retinopathy, diabetic macular edema (DME), clinically significant macular edema (CSME), cystoid macular edema (CME), CME after cataract extraction, CME induced by cryotherapy, CME induced by uveitis, endophthalmitis, CME after vascular occlusion (e.g., central retinal vein occlusion, branch retinal vein occlusion or hemi - retinal vein occlusion), retinal edema, complications associated with cataract surgery in diabetic retinopathy, hypertensive retinopathy, retinal trauma, atrophic and exudative age - related macular degeneration (AMD), polypoidal choroidal vasculopathy (PCV), choroidal neovascularization (CNV; e.g., non - exudative choroidal neovascularization), hereditary angioedema (HAE), acute respiratory distress syndrome (ARDS), hemorrhage and edema after stroke, e.g., cerebral edema after stroke, vascular dementia, Alzheimer's disease, fibrotic diseases, colitis, arthritis and kidney injury. Accordingly, the compounds and pharmaceutical compositions of the invention are particularly suitable for eye diseases, including diabetic retinopathy, proliferative and non - proliferative retinopathy, diabetic macular edema (DME), retinal vein occlusion, age - related macular degeneration (AMD), polypoidal choroidal vasculopathy (PCV) and choroidal neovascularization (CNV; e.g., non - exudative choroidal neovascularization).

[0071] Furthermore, the compounds and pharmaceutical compositions according to the present invention are particularly suitable for the treatment of edema such as hereditary angioedema (HAE) and cerebral edema after stroke. In particular, the compounds and pharmaceutical compositions according to the present invention are suitable for the treatment of diabetic retinopathy, proliferative and non-proliferative retinopathy, diabetic macular edema (DME), age-related macular degeneration (AMD), polypoidal choroidal vasculopathy (PCV), choroidal neovascularization (CNV), hereditary angioedema (HAE), and cerebral edema after stroke. The compounds and pharmaceutical compositions according to the present invention are most particularly suitable for the treatment of diabetic macular edema (DME), exudative age-related macular degeneration (AMD), non-exudative choroidal neovascularization (CNV), hereditary angioedema (HAE), and cerebral edema after stroke.

[0072] For example, the compounds and pharmaceutical compositions according to the present invention are particularly suitable for the prevention of diabetic macular edema (DME), exudative age-related macular degeneration (AMD), hereditary angioedema (HAE) and cerebral edema after stroke, and the prevention of the conversion from non-exudative choroidal neovascularization (neCNV) to exudative choroidal neovascularization (eCNV). The dosage range of the compound of formula (I) that can be administered per day is usually 0.01 to 10 mg per kg of body weight. The actual therapeutically effective amount or therapeutic dosage will, of course, depend on factors known to those skilled in the art such as the age and weight of the patient, the route of administration and the severity of the disease. In any case, the compound or composition will be administered in a dosage and manner capable of delivering a therapeutically effective amount based on the particular condition of the patient. The present compounds and compositions can be administered by oral, intravitreal, transdermal, inhalation, parenteral or sublingual routes, including any combination with one or more additional therapeutic agents according to the present invention. Among the possible methods of administration, oral administration or intravitreal administration is preferred. In the case of intravitreal injection, the preferred dosage should not exceed 5 mg per eye.

[0073] The patient to be treated is preferably a mammal, most preferably a human patient. Accordingly, in another aspect, the present invention provides a compound of formula (I) and its tautomers (including pharmaceutically acceptable salts thereof) for use as a medicament. In another aspect, the present invention provides a method of treating a disease or condition mediated by unwanted plasma kallikrein activity or in which inhibition of plasma kallikrein is beneficial in a patient in need thereof. Similarly, the present invention provides a compound of formula (I) and / or its tautomer or a pharmaceutically acceptable salt thereof for use in a method of treating a disease or condition mediated by unwanted plasma kallikrein activity or in which inhibition of plasma kallikrein is beneficial in a patient in need thereof. Similarly, the present invention provides the use of a compound of formula (I) and / or its tautomer or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for use in a method of treating a disease or condition mediated by unwanted plasma kallikrein activity or in which inhibition of plasma kallikrein is beneficial in a patient in need thereof.

[0074] Similarly, the present invention provides the use of a compound of formula (I) and / or its tautomer or a pharmaceutically acceptable salt thereof in a method of treating a disease or condition mediated by unwanted plasma kallikrein activity or in which inhibition of plasma kallikrein is beneficial in a patient in need thereof. According to one embodiment, the method of treatment comprises administering to the patient one or more compounds of formula (I), and / or its tautomer or a pharmaceutically acceptable salt thereof, preferably administering to the patient a therapeutically effective amount of one or more compounds of formula (I), and / or its tautomer or a pharmaceutically acceptable salt thereof. According to another embodiment, the method of treatment comprises administering to the patient a pharmaceutical composition according to the present invention. According to one embodiment, diseases or conditions mediated by undesirable plasma kallikrein activity or for which inhibition of plasma kallikrein is beneficial are selected from ophthalmic indications such as diabetic retinopathy, proliferative and non-proliferative retinopathy, diabetic macular edema (DME), age-related macular degeneration (AMD), polypoidal choroidal vasculopathy (PCV), and choroidal neovascularization (CNV).

[0075] According to another embodiment, diseases or conditions mediated by undesirable plasma kallikrein activity or for which inhibition of plasma kallikrein is beneficial are selected from edema-related diseases such as hereditary angioedema (HAE) and cerebral edema after stroke. According to another embodiment, diseases or conditions mediated by undesirable plasma kallikrein activity or for which inhibition of plasma kallikrein is beneficial are selected from diabetic complications such as diabetic retinopathy and retinal vascular permeability associated with diabetic macular edema. According to one embodiment, the patient is a human patient.

[0076] Pharmaceutical composition In another aspect of the invention, it is described that the compounds of the invention or pharmaceutically acceptable salts thereof may be used as the active ingredient in a pharmaceutical composition. Suitable preparations for administering the compounds of the invention, which may be combined with one or more additional therapeutic agents, will be apparent to those skilled in the art and include, for example, tablets, pills, capsules, suppositories, lozenges, troches, solutions, syrups, elixirs, sachets, injections, inhalants, and powders. Oral formulations, particularly solid forms such as tablets or capsules, are preferred. In the case of intravitreal injection, a solution is preferred. The content of the pharmaceutically active compound is advantageously in the range of 0.1 to 90% by weight, for example 1 to 70% by weight, of the composition as a whole.

[0077] Suitable tablets can be obtained, for example, by mixing one or more compounds according to formula (I) with known excipients, such as inert diluents, carriers, disintegrants, adjuvants, surfactants, binders and / or lubricants. The tablets may also consist of several layers. Specific excipients, carriers and / or diluents suitable for the desired preparation will be familiar to those skilled in the art based on the knowledge of experts. Preferred are those suitable for the particular formulation and method of administration desired. The preparations or formulations according to the invention can be prepared by methods known per se to those skilled in the art, such as, for example, by mixing or combining at least one compound of formula (I) according to the invention or a pharmaceutically acceptable salt of such a compound with one or more excipients, carriers and / or diluents. Accordingly, according to another aspect of the invention, there is provided a pharmaceutical composition comprising one or more compounds of formula (I) and / or their tautomers, or pharmaceutically acceptable salts thereof, which may also contain one or more inert carriers and / or diluents together.

[0078] Similarly, there is provided a pharmaceutical composition comprising one or more of the above compounds or pharmaceutically acceptable salts thereof for use in a method of treating a disease or condition mediated by undesirable plasma kallikrein activity or in which inhibition of plasma kallikrein is beneficial in a patient in need thereof, which may also contain one or more inert carriers and / or diluents together. In particular, the present invention provides a pharmaceutical composition according to the invention for use in a method of treating ophthalmic indications such as diabetic retinopathy, proliferative and non-proliferative retinopathy, diabetic macular edema (DME), age-related macular degeneration (AMD), polypoidal choroidal vasculopathy (PCV) and choroidal neovascularization (CNV), as well as edema-related diseases such as hereditary angioedema (HAE) and cerebral edema after stroke. Furthermore, the present invention relates to the use of a pharmaceutical composition according to the invention for treating a disease or condition mediated by undesirable plasma kallikrein activity in a patient, preferably a human. Similarly, the present invention relates to the use of a pharmaceutical composition according to the present invention for treating a disease or condition in a patient, preferably a human, in which inhibition of plasma kallikrein is beneficial. According to another embodiment, there is provided a pharmaceutical composition comprising one or more compounds of formula (I) and / or their tautomers, or pharmaceutically acceptable salts thereof, and one or more additional therapeutic agents, which may together comprise one or more inert carriers and / or diluents. Preferably, the composition comprises a compound of formula (I) and / or its tautomer, or a pharmaceutically acceptable salt thereof, and one or more additional therapeutic agents.

[0079] Combination therapy The compounds of the present invention may further be combined with one or more, preferably one, additional therapeutic agent. According to one embodiment, the additional therapeutic agent is, for example, a therapeutic agent useful for treating a disease or condition described previously herein that is particularly related to a metabolic disease or condition such as diabetes, obesity, diabetic complications, hypertension, hyperlipidemia, or a therapeutic agent useful for treating an eye disease, selected from the group of therapeutic agents. Additional therapeutic agents suitable for such combinations include, in particular, those that enhance, for example, the therapeutic action of one or more active substances and / or enable a reduction in the dosage of one or more active substances, with respect to one of the indicated indications. Thus, the compounds of the present invention can be combined with one or more additional therapeutic agents selected from the group consisting of anti-diabetic agents, agents for treating overweight and / or obesity, agents for treating hypertension, heart failure and / or atherosclerosis, and agents for treating eye diseases.

[0080] Antidiabetic agents include, for example, metformin, sulfonylureas, nateglinide, repaglinide, thiazolidinediones, PPAR-(alpha, gamma or alpha / gamma) agonists or modulators, alpha-glucosidase inhibitors, DPPIV inhibitors, SGLT2 inhibitors, insulin and insulin analogs, GLP-1 and GLP-1 analogs or amylin and amylin analogs, cycloset, 11β-HSD inhibitors. Other suitable combination partners are substances that affect deregulated glucose production in the liver, such as inhibitors of glucose-6-phosphatase or fructose-1,6-bisphosphatase, glycogen phosphorylase, glucagon receptor antagonists, and phosphoenolpyruvate carboxykinase, inhibitors of glycogen synthase kinase or pyruvate dehydrogenase kinase, alpha2-antagonists, CCR-2 antagonists or glucokinase activators, and inhibitors of protein tyrosine phosphatase 1. For example, cholesterol absorption inhibitors such as bile acid-binding substances such as HMG-CoA-reductase inhibitors, fibrates, nicotinic acid and its derivatives, PPAR-(alpha, gamma or alpha / gamma) agonists or modulators, PPAR-delta agonists, ACAT inhibitors, or inhibitors of the ileal bile acid transporter, MTP inhibitors, or HDL-raising compounds such as CETP inhibitors or ABC1 regulators, one or more lipid-lowering agents are also suitable as combination partners.

[0081] Therapeutic agents for treating overweight and / or obesity include, for example, antagonists of the cannabinoid 1 receptor, MCH-1 receptor antagonists, MC4 receptor agonists, NPY5 or NPY2 antagonists, beta3-agonists, leptin or leptin mimetics, agonists of the 5HT2c receptor. Therapeutic agents for the treatment of hypertension, chronic heart failure and / or atherosclerosis are, for example, A-II antagonists or ACE inhibitors, ECE inhibitors, diuretics, β-blockers, Ca-antagonists, centrally acting antihypertensives, antagonists of the alpha-2-adrenergic receptor, inhibitors of neutral endopeptidase, platelet aggregation inhibitors, and others or combinations thereof are suitable. Angiotensin II receptor antagonists are preferably used for the treatment or prevention of hypertension and complications of diabetes, often in combination with a diuretic such as hydrochlorothiazide.

[0082] Therapeutic agents for the treatment of eye diseases can include, for example, corticosteroids administered intravitreally, anti-VEGF therapy administered intravitreally, anti-Ang2 inhibitors, dual anti-VEGF / anti-Ang2 inhibitors, anti-PDGF, dual anti-VEGF / anti-PDGF, VAP-1 (AOC3) inhibitors, complement inhibitors (e.g., complement factor 3, 5, B and D inhibitors), bradykinin receptor 1 antagonists, CCR-2 antagonists. Additional treatments for eye diseases can include laser photocoagulation therapy. Preferably, the compounds of the present invention, and / or pharmaceutical compositions that may contain the compounds of the present invention in combination with one or more additional therapeutic agents, are administered together with exercise and / or diet. The dosage of the above combination partners is usually from 1 / 5 of the usually recommended minimum dose to 1 / 1 of the usually recommended dose. The use of the compounds according to the invention in combination with additional therapeutic agents can be carried out simultaneously or with a time difference.

[0083] Both the compounds according to the present invention and one or more additional therapeutic agents may be present together in one formulation, such as a tablet or capsule, or separately in two identical or different formulations, for example, as a so-called kit of parts. Accordingly, in another aspect, the present invention relates to a pharmaceutical composition comprising one or more compounds according to the present invention, and one or more additional therapeutic agents described hereinbefore and hereinafter in this specification, which may together comprise one or more inert carriers and / or diluents. In another aspect, the present invention provides a method of treating a disease or condition mediated by undesirable plasma kallikrein activity or in which inhibition of plasma kallikrein is beneficial in a patient in need thereof, the method comprising administering to the patient one or more compounds of formula (I), and / or a tautomer or pharmaceutically acceptable salt thereof, in combination with one or more additional therapeutic agents described hereinbefore and hereinafter in this specification. Preferably, there is provided a method comprising administering to the patient a therapeutically effective amount of one or more compounds of formula (I), and / or a tautomer or pharmaceutically acceptable salt thereof, in combination with a therapeutically effective amount of one or more additional therapeutic agents described hereinbefore and hereinafter in this specification.

[0084] Similarly, the present invention provides a compound of formula (I), and / or a tautomer or pharmaceutically acceptable salt thereof, in combination with one or more additional therapeutic agents described hereinbefore or hereinafter in this specification for use in a method of treating a disease or condition mediated by undesirable plasma kallikrein activity or in which inhibition of plasma kallikrein is beneficial in a patient in need thereof. Similarly, the present invention provides the use of a compound of formula (I), and / or a tautomer or pharmaceutically acceptable salt thereof, in combination with one or more additional therapeutic agents described hereinbefore or hereinafter in this specification in the manufacture of a medicament for use in a method of treating a disease or condition mediated by undesirable plasma kallikrein activity or in which inhibition of plasma kallikrein is beneficial in a patient in need thereof. Similarly, the present invention provides for the use of a compound of formula (I) and / or a tautomer or a pharmaceutically acceptable salt thereof in combination with one or more additional therapeutic agents described hereinbefore or hereinafter in a method of treating a disease or condition in a patient in which undesirable plasma kallikrein activity is mediated or in which inhibition of plasma kallikrein would be beneficial.

[0085] According to one embodiment, the method of treatment comprises administering to a patient one or more compounds of formula (I), and / or a tautomer or a pharmaceutically acceptable salt thereof, in combination with one or more additional therapeutic agents described hereinbefore and hereinafter. Preferably, the method of treatment comprises administering to a patient a therapeutically effective amount of one or more compounds of formula (I), and / or a tautomer or a pharmaceutically acceptable salt thereof, in combination with a therapeutically effective amount of one or more additional therapeutic agents described hereinbefore and hereinafter. According to another embodiment, the method of treatment comprises administering to a patient a pharmaceutical composition comprising one or more compounds according to the invention and one or more additional therapeutic agents described hereinbefore and hereinafter, which may together comprise one or more inert carriers and / or diluents. According to one embodiment, the one or more additional therapeutic agents are selected from anti-diabetic agents, agents for the treatment of overweight and / or obesity, agents for the treatment of hypertension, heart failure and / or atherosclerosis, and agents for the treatment of eye diseases, in particular such agents specifically mentioned above.

[0086] According to one embodiment, diseases or conditions mediated by undesirable plasma kallikrein activity or for which inhibition of plasma kallikrein is beneficial are selected from ophthalmic indications such as diabetic retinopathy, proliferative and non-proliferative retinopathy, diabetic macular edema (DME), age-related macular degeneration (AMD), polypoidal choroidal vasculopathy (PCV), and choroidal neovascularization (CNV), from edema-related diseases such as hereditary angioedema (HAE) and cerebral edema after stroke, or from diabetic complications such as retinal vascular permeability associated with diabetic retinopathy and diabetic macular edema. According to one embodiment, the patient is a human patient. Other features and advantages of the invention will become apparent from the following more detailed examples, which illustrate the principles of the invention by way of example.

Examples

[0087] Examples and experimental data The following examples are for illustrative purposes only of the invention and are not intended to limit the scope of the invention in any way. Abbreviations Ac Acetyl ACN Acetonitrile AMC 7-Amino-4-methylcoumarin Boc tert-Butyloxycarbonyl BSA Bovine serum albumin Bzl Benzyl d Days DABCO 1,4-Diazabicyclo[2.2.2]octane DAD Diode array detector DBAD Di-tert-butyl azodicarboxylate DBU 1,8-Diazabicyclo[5.4.0]undec-7-ene DBN 1,5-Diazabicyclo[4.3.0]nona-5-ene DCM Dichloromethane DIPEA N,N-Diisopropylethylamine DMEM Dulbecco's Modified Eagle Medium DMF N,N-Dimethylformamide DMSO Dimethyl sulfoxide EDTA Ethylenediaminetetraacetate ESI Electrospray ionization (MS) EtOAc Ethyl acetate EtOH Ethanol h hour HATU O-(7-Azabenzotriazol-1-yl)-N,N,N’,N’-tetramethyluronium-hexafluorophosphate HPLC High performance liquid chromatography HPLC-MS Coupled high performance liquid chromatography - mass spectrometry LC Liquid chromatography LC-MS Coupled liquid chromatography - mass spectrometry LG Leaving group M Molar concentration (mol / L) MeOH Methanol min minute MS Mass spectrometry NADPH Nicotinamide adenine dinucleotide phosphate NMP N-Methyl-2-pyrrolidone NMR Nuclear magnetic resonance PET Polyethylene terephthalate PyBop (Benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate R f Retention factor RFU Relative fluorescence unit RP Reverse phase rt Room temperature t R Retention time (HPLC / LC) TBTU O-(Benzotriazol-1-yl)-N,N,N’,N’-tetramethyluronium tetrafluoroborate TFA Trifluoroacetic acid THF Tetrahydrofuran UV Ultraviolet

[0088] The terms "ambient temperature" and "room temperature" are used interchangeably and represent a temperature of about 20 °C, for example, a temperature in the range of 15 - 25 °C. Generally, for the prepared compounds, 1 1H-NMR and / or mass spectra were obtained. Unless otherwise specified, compounds containing chiral centers have the stereochemistry shown. The stereochemical assignments are made either by use of chiral starting materials of known stereochemistry, by stereoselective synthesis of known stereochemistry, or by biological activity.

[0089] [Table 6]

[0090] [Table 7]

[0091] [Table 8]

[0092] Synthesis of intermediates: The starting materials and intermediates used in the process leading to the compounds according to the invention are either commercially available or they can be prepared by the methods described below (or methods similar or analogous thereto) or from the literature, for example, by methods already known to the person skilled in the art from WO2017 / 072020, WO2017 / 072021 and WO2018 / 192866, the entire contents of which are incorporated herein by reference.

[0093] Intermediate 1 (6R)-3-Methyl-2H,4H,5H,6H-cyclopenta[c]pyrazol-6-amine; semi-(2S,3S)-2,3-bis(4-methylbenzoyloxy)butanedioate [Chemical formula]

[0094] Step 1: Ethyl 4-(3-ethoxy-3-oxopropyl)-5-methyl-1H-pyrazole-3-carboxylate Under an argon atmosphere, a mixture consisting of ethyl 4-iodo-5-methyl-1H-pyrazole-3-carboxylate (220 g), 3,3-diethoxyprop-1-ene (112 g), palladium(II) acetate (Pd(OAc) 2 , 35.3 g), tetrabutylammonium chloride (240 g) and DIPEA (203 g) in DMF (900 mL) is stirred at 110 °C for 2 hours and at 120 °C for 1 hour. After cooling to about 67 °C, EtOAc (1.5 L) is added. The mixture is cooled to room temperature and filtered through celite, and the filter cake is washed with EtOAc (2x). The combined filtrate is washed with semi-saturated aqueous NaCl solution (3x) and water (4x). After drying (MgSO 4 ), the mixture is concentrated in vacuo to give a crude product, which is used directly in the next step. Mass spectrum (ESI + ): m / z = 255 [M+H] + .

[0095] Step 2: Ethyl 3-methyl-6-oxo-2H,4H,5H,6H-cyclopenta[c]pyrazole-5-carboxylate Under an argon atmosphere, a solution of ethyl 4-(3-ethoxy-3-oxopropyl)-5-methyl-1H-pyrazole-3-carboxylate (191 g) in THF (500 mL) is added between 41 - 64 °C to a solution of NaN(Si(CH 3 )([[]]END]] 3 )([[]]END]] 2 ) (2 M in THF, 1.3 L). The mixture is stirred at 60 °C for 15 minutes and at room temperature for 1 hour. Next, the mixture is poured into a mixture consisting of aqueous HCl (6 N, 851 mL), ice (1 kg) and 2-methyl-tetrahydrofuran (2.5 L). After stirring for 10 minutes, the phases are separated. The organic phase is concentrated in vacuo to give a crude product, which is used directly in the next step. Mass spectrum (ESI + ): m / z = 209 [M+H] + .

[0096] Step 3: 3-Methyl-2H,4H,5H,6H-cyclopenta[c]pyrazol-6-one A mixture consisting of ethyl 3-methyl-6-oxo-2H,4H,5H,6H-cyclopenta[c]pyrazole-5-carboxylate (363 g) in 1,4-dioxane (1.5 L) and water (110 mL) is heated to 90 °C for 22 h. The mixture is concentrated in vacuo to give the crude product, which is used directly in the next step. Mass spectrum (ESI + ): m / z = 137 [M+H] + .

[0097] Step 4: N-[3-Methyl-2H,4H,5H,6H-cyclopenta[c]pyrazol-6-ylidene]hydroxylamine; hydrochloride A solution of 3-methyl-2H,4H,5H,6H-cyclopenta[c]pyrazol-6-one (294 g) in EtOH (800 mL) is heated to 65 °C and treated with hydroxylamine hydrochloride (52.5 g). The mixture is stirred at 60 °C for 2 h and at room temperature for 12 h. The precipitate is collected and washed with EtOH and tert-butyl-methyl-ether. The crude product is dried in vacuo and used directly in the next step. Mass spectrum (ESI + ): m / z = 152 [M+H] + .

[0098] Step 5: (6R)-3-Methyl-2H,4H,5H,6H-cyclopenta[c]pyrazol-6-amine; semi-(2S,3S)-2,3-bis(4-methylbenzoyloxy)-butanedioate N-[3-Methyl-2H,4H,5H,6H-cyclopenta[c]pyrazol-6-ylidene]hydroxylamine; hydrochloride (85 g) is suspended in MeOH (2.2 L) and treated with (1R)-1-phenylethan-1-amine (165 g) and stirred at 40 °C for 30 min. 20% Pd(OH) on carbon 2(18 g) is added and the mixture is stirred under a hydrogen atmosphere (21 bar) at 65 °C for 8 hours and at 100 °C for 12 hours. The mixture is cooled to room temperature, filtered through celite, and concentrated in vacuo. The residue is dissolved in isopropanol (1.4 L) and water (70 mL), heated to 60 °C, and treated with K 2 CO 3 (94 g). The mixture is stirred at 60 °C for 10 minutes and at 55 °C for 50 minutes, filtered, and the filter cake is washed with isopropanol. The combined filtrates are concentrated in vacuo. The residue is dissolved in isopropanol (100 mL), heated to 60 °C, and treated with a solution of (2S,3S)-2,3-bis(4-methylbenzoyloxy)butanedioic acid (79 g) in isopropanol (700 mL) and water (40 mL). The mixture is heated to 35 °C and filtered. The filtrate is stirred for 30 minutes, heated to 60 °C for 1 hour, and then stirred at room temperature for 12 hours. The precipitate is collected by filtration and washed with isopropanol and tert-butyl-methyl-ether. The crude product is dried in vacuo and used directly in the next step. Mass spectrum (ESI + ): m / z = 138 [M+H] + .

[0099] Intermediate 2 6-{5-Azaspiro[2.3]hexan-5-yl}-2-chloropyridine-3-carbaldehyde [Chemical formula] A mixture consisting of 2,6-dichloropyridine-3-carbaldehyde (10 g), 5-azaspiro[2.3]hexane trifluoroacetate (16 g), and DIPEA (40 mL) in DMF (40 mL) is heated to 40 °C for 12 hours under an argon atmosphere. The mixture is cooled, concentrated, partitioned between water and DCM, and the phases are separated. The organic phase is washed with brine, dried (MgSO 4 ) and concentrated, and the residue is subjected to chromatography (petroleum ether / EtOAc 90:10 → 70:30) on silica gel to give the title compound. LC (Method 2): t R=1.03 min; Mass spectrum (ESI+): m / z = 223 [M+H] + 。

[0100]

Table 9

[0101]

Table 10

[0102]

Table 11

[0103] Intermediate 3 (6-{5-Azaspiro[2.3]hexan-5-yl}-2-chloropyridin-3-yl)methanol

Chem.

[0104]

Table 12

[0105]

Table 13

[0106]

Table 14

[0107] Intermediate 4 Ethyl 1-[(6-{5-azaspiro[2.3]hexan-5-yl}-2-chloropyridin-3-yl)methyl]-1H-pyrazole-4-carboxylate

Chemical formula

[0108]

Table 15-1

Table 15-2

[0109]

Table 16

[0110]

Table 17

[0111] Intermediate 5 Ethyl 1-[(6-{5-azaspiro[2.3]hexan-5-yl}-2-ethenylpyridin-3-yl)methyl]-1H-pyrazole-4-carboxylate

Chemical formula

[0112]

Table 18

[0113]

Table 19

[0114] Intermediate 6 Ethyl 1-[(6-{5-azaspiro[2.3]hexan-5-yl}-2-formylpyridin-3-yl)methyl]-1H-pyrazole-4-carboxylate

Chem.

[0115]

Table 20

[0116]

Table 21

[0117]

Table 22

[0118] Intermediate 7 Ethyl 1-[(6-{5-azaspiro[2.3]hexan-5-yl}-2-(difluoromethyl)pyridin-3-yl)methyl]-1H-pyrazole-4-carboxylate

Chem.

[0119]

Table 23

[0120]

Table 24

[0121] Intermediate 8 1-[(6-{5-Azaspiro[2.3]hexan-5-yl}-2-(difluoromethyl)pyridin-3-yl)methyl]-1H-pyrazole-4-carboxylic acid

Chem.

[0122]

Table 25-1

Table 25-2

[0123]

Table 26

[0124]

Table 27

[0125] Intermediate 9 Ethyl 2-chloro-4-[(1E)-2-phenylethenyl]pyrimidine-5-carboxylate

Chem.

[0126] Intermediate 10 Ethyl 2-{3-azabicyclo[3.1.0]hexan-3-yl}-4-[(1E)-2-phenylethenyl]pyrimidine-5-carboxylate

Chemical formula

[0127]

Table 28

[0128]

Table 29

Table 30

[0129] Intermediate 11 (2-{3-Azabicyclo[3.1.0]hexan-3-yl}-4-[(1E)-2-phenylethenyl]pyrimidin-5-yl)methanol

Chem.

[0130] Intermediate 12 Ethyl 1-[(2-{3-azabicyclo[3.1.0]hexan-3-yl}-4-[(1E)-2-phenylethenyl]pyrimidin-5-yl)methyl]-1H-pyrazole-4-carboxylate

Chem.

[0131] Intermediate 13 Ethyl 1-[(6-{3-azabicyclo[3.1.0]hexan-3-yl}-2-(difluoromethyl)pyridin-3-yl)methyl]-1H-1,2,3-triazole-4-carboxylate

Chem.

[0132] Intermediate 14 Ethyl 1-[(6-{5-azaspiro[2.3]hexan-5-yl}-2-ethenylpyridin-3-yl)methyl]-1H-imidazole-4-carboxylate [Chemical formula] A stir bar, ethyl 1-[(6-{5-azaspiro[2.3]hexan-5-yl}-2-chloropyridin-3-yl)methyl]-1H-imidazole-4-carboxylate (2.6 g), vinylboronic acid pinacol ester (1.4 mL), Na 2 CO 3 (1 M aqueous solution, 18.6 mL) and 1,4-dioxane (40 mL) were charged into a microwave vial, and argon was vigorously flowed through for 5 minutes. 1,1'-Bis(diphenylphosphino)ferrocene palladium(II) dichloride (Pd(dppf)Cl 2, 304 mg) is added, the vial is sealed, and this mixture is stirred at 100 °C for 6 hours. After cooling this mixture to room temperature, it is partitioned between water and EtOAc. The aqueous phase is extracted twice with EtOAc. The combined organic phases are washed with brine, dried (MgSO 4 ) and concentrated. The residue is subjected to chromatography (petroleum ether / EtOAc 50:50 → 0:100) on silica gel to give the title compound. LC (Method 1): t R = 1.01 min; mass spectrum (ESI+): m / z = 339 [M+H] + .

[0133]

Table 31

[0134]

Table 32

[0135]

Table 33

[0136] Intermediate 15 (6-{3-Azabicyclo[3.1.0]hexan-3-yl}-4-(trifluoromethyl)pyridin-3-yl)methanol

Chem.

[0137] Intermediate 16 1-[(6-{3-Azabicyclo[3.1.0]hexan-3-yl}-2-(trifluoromethyl)pyridin-3-yl)methyl]-N-[(6R)-3-methyl-2-{[2-(trimethylsilyl)ethoxy]methyl}-2H,4H,5H,6H-cyclopenta[c]pyrazol-6-yl]-1H-pyrazole-4-carboxamide

Chemical formula

[0138] Intermediate 17 (6R)-3-Methyl-2-{[2-(trimethylsilyl)ethoxy]methyl}-2H,4H,5H,6H-cyclopenta[c]pyrazol-6-amine [Chemical Structure]

[0139] Step 1: Methyl 5-methyl-1H-pyrazole-3-carboxylate Thionyl chloride (58 mL) is added dropwise to a solution of 5-methyl-1H-pyrazole-3-carboxylic acid (45 g) in MeOH (450 mL). After the addition, the mixture is stirred at room temperature for 16 hours. The mixture is concentrated in vacuo. The residue is dissolved in EtOAc and washed successively with saturated NaHCO 3 aqueous solution and brine. After drying (MgSO 4 ), the mixture is concentrated in vacuo to give the title compound. LC (Method 1): t R = 0.64 min; Mass spectrum (ESI + ): m / z = 141 [M+H] + .

[0140] Step 2: Methyl 5-methyl-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrazole-3-carboxylate Sodium hydride (60% in mineral oil, 16.8 g) is added portionwise to DMF (470 mL). The mixture is stirred for 10 minutes, cooled to 0 °C, and treated with a solution of methyl 5-methyl-1H-pyrazole-3-carboxylate (46.9 g) in DMF (470 mL) added dropwise. After stirring for 20 minutes, [2-(chloromethoxy)ethyl]trimethylsilane (SEM-Cl, 77.7 mL) is added dropwise. The mixture is stirred for 2 hours, diluted with EtOAc, and washed successively with water and brine. After drying (MgSO 4)Subsequently, this mixture is concentrated in vacuo, and the residue is chromatographed on silica gel with petroleum ether / EtOAc 2:1. Evaporation of the solvent in vacuo gives the title compound, which is used directly in the next step.

[0141] Step 3: Methyl 4-iodo-5-methyl-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrazole-3-carboxylate To a solution of methyl 5-methyl-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrazole-3-carboxylate (94.4 g) in ACN (1.4 L) are added TFA (2.7 mL) and N-iodosuccinimide (94.2 g). The mixture is stirred for 48 h, diluted with EtOAc, and washed successively with water, saturated Na 2 S 2 O 3 aqueous solution and brine. After drying (MgSO 4 ), this mixture is concentrated in vacuo, and the residue is chromatographed on silica gel with petroleum ether / EtOAc 2:1. Evaporation of the solvent in vacuo gives the title compound, which contains approximately 15% of the positional isomer methyl 4-iodo-3-methyl-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrazole-5-carboxylate. LC (Method 1): t R = 1.17 min; mass spectrum (ESI + ): m / z = 397 [M + H] + .

[0142] Step 4: Methyl 4-[(1E)-3-methoxy-3-oxoprop-1-en-1-yl]-5-methyl-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrazole-3-carboxylate Methyl 4-iodo-5-methyl-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrazole-3-carboxylate (44 g), methyl acrylate (15 mL) and N-methyldicyclohexylamine (35 mL) are dissolved in dimethylacetamide (430 mL) and water (110 mL). The mixture is purged with argon for 10 minutes. Dichlorobis(tri-o-tolylphosphine)palladium(II) (PdCl 2 [P(o-Tol) 3 2 , 2.6 g) is added and the mixture is stirred at 85 °C for 2 hours. Next, the mixture is diluted with EtOAc and washed successively with 1M H 3 PO 4 aqueous solution and brine. After drying (MgSO 4 ), the mixture is concentrated in vacuo and the residue is subjected to chromatography on silica gel (petroleum ether / EtOAc 95:5 → 50:50) to give the title compound. LC (Method 1): t R = 1.13 min; mass spectrum (ESI + ): m / z = 355 [M+H] + .

[0143] Step 5: Methyl 4-(3-methoxy-3-oxopropyl)-5-methyl-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrazole-3-carboxylate A mixture consisting of methyl 4-[(1E)-3-methoxy-3-oxoprop-1-en-1-yl]-5-methyl-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrazole-3-carboxylate (38.6 g) and 10% palladium on carbon (5.8 g) in EtOAc (580 mL) is shaken at room temperature for 3 hours under a hydrogen atmosphere (3 bar). The mixture is filtered and the filtrate is concentrated to give the title compound. LC (Method 1): t R = 1.10 min; mass spectrum (ESI + ): m / z = 357 [M+H] + . ​

[0144] Step 6: Methyl 3-methyl-6-oxo-2-{[2-(trimethylsilyl)ethoxy]methyl}-2H,4H,5H,6H-cyclopenta[c]pyrazole-5-carboxylate A solution of methyl 4-(3-methoxy-3-oxopropyl)-5-methyl-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrazole-3-carboxylate (37.8 g) in THF was cooled to 0 °C and treated with NaN(Si(CH 3 ) 3 ) 2 (40% in THF; 105 mL) and stirred for 15 minutes. The mixture was poured into 1 M H 3 PO 4 aqueous solution under ice-cooling and vigorous stirring. The organic phase was separated, washed with brine and dried (MgSO 4 ). Evaporation of the solvent in vacuo gave the title compound. LC (Method 1): t R = 1.06 min; mass spectrum (ESI + ): m / z = 325 [M+H] + .

[0145] Step 7: 3-Methyl-2-{[2-(trimethylsilyl)ethoxy]methyl}-2H,4H,5H,6H-cyclopenta[c]pyrazol-6-one A solution of methyl 3-methyl-6-oxo-2-{[2-(trimethylsilyl)ethoxy]methyl}-2H,4H,5H,6H-cyclopenta[c]pyrazole-5-carboxylate (34.9 g) in 1,4-dioxane (350 mL) and water (9 mL) was heated under reflux for 12 hours. Evaporation of the solvent in vacuo and chromatography of the residue on silica gel (petroleum ether / EtOAc 80:20 → 40:60) gave the title compound. LC (Method 1): t R = 1.05 min; mass spectrum (ESI + ): m / z = 267 [M+H] + .

[0146] Step 8: (6S)-3-Methyl-2-{[2-(trimethylsilyl)ethoxy]methyl}-2H,4H,5H,6H-cyclopenta[c]pyrazol-6-ol To a solution of triethylamine (27 mL) in DCM (260 mL) is added formic acid (11 mL) at 0 °C. The mixture is warmed to room temperature and 3-methyl-2-{[2-(trimethylsilyl)ethoxy]methyl}-2H,4H,5H,6H-cyclopenta[c]pyrazol-6-one (26 g) is added. After purging with argon for 10 minutes, [N-[(1S,2S)-2-(amino-κN)-1,2-diphenylethyl]-4-methylbenzenesulfonamidato-κN]chloro-[(1,2,3,4,5,6-η)-1,3,5-trimethylbenzene]-ruthenium (RuCl[(S,S)-TsDPEN](mesitylene); 0.5 g) is added and the mixture is stirred at room temperature for 48 hours. Next, the mixture is treated with 1 M NaHCO 3 aqueous solution under vigorous stirring. The phases are separated and the aqueous phase is extracted with DCM. The combined organic phases are washed with water and brine. After drying (MgSO 4 ), the solvent is evaporated in vacuo and the residue is subjected to chromatography on silica gel (DCM / MeOH 98:2 → 90:10) to give the title compound with an enantiomeric excess (ee) of 84%. LC (Method 1): t R = 0.99 min; mass spectrum (ESI + ): m / z = 269 [M+H] + .

[0147] Step 9: (6R)-6-Azido-3-methyl-2-{[2-(trimethylsilyl)ethoxy]methyl}-2H,4H,5H,6H-cyclopenta[c]pyrazole Under an argon atmosphere, DBU (16 mL) is added to a toluene (250 mL) solution of (6S)-3-methyl-2-{[2-(trimethylsilyl)ethoxy]methyl}-2H,4H,5H,6H-cyclopenta[c]pyrazol-6-ol (25.5 g). The mixture is cooled to 0 °C, and diphenylphosphoryl azide (22 mL) is added dropwise over 1 hour. The mixture is stirred for 12 hours while warming to room temperature. Next, MeOH (25 mL) is added, and the mixture is stirred for 1 hour. The mixture is washed twice with water, dried (MgSO 4 ) and concentrated in vacuo. The residue is chromatographed on Al 2 O 3 (DCM) to give the title compound. LC (Method 1): t R = 1.15 min; mass spectrum (ESI + ): m / z = 294 [M+H] + .

[0148] Step 10: (6R)-3-methyl-2-{[2-(trimethylsilyl)ethoxy]methyl}-2H,4H,5H,6H-cyclopenta[c]pyrazol-6-amine A mixture consisting of (6R)-6-azido-3-methyl-2-{[2-(trimethylsilyl)ethoxy]methyl}-2H,4H,5H,6H-cyclopenta[c]pyrazole (19.7 g) and 10% palladium on carbon (3 g) in EtOH (200 mL) is shaken at room temperature for 12 hours under a hydrogen atmosphere (3 bar). The mixture is filtered, and the filtrate is concentrated to give the title compound. LC (Method 1): t R = 0.80 min; mass spectrum (ESI + ): m / z = 268 [M+H] + .

[0149] Examples of synthesis: (Example 1) 1-[(6-{5-Azaspiro[2.3]hexan-5-yl}-2-(difluoromethyl)pyridin-3-yl)methyl]-N-[(6R)-3-methyl-2H,4H,5H,6H-cyclopenta[c]pyrazol-6-yl]-1H-pyrazole-4-carboxamide

Chem.

[0150]

Table 34-1

Table 34-2

[0151]

Table 35

[0152] (Example 13) 1-[(6-{3-azabicyclo[3.1.0]hexan-3-yl}-2-(trifluoromethyl)pyridin-3-yl)methyl]-N-[(6R)-3-methyl-2H,4H,5H,6H-cyclopenta[c]pyrazol-6-yl]-1H-pyrazole-4-carboxamide

[0153]

Chem.

Claims

1. A compound of formula (I) 【Chemical 1】 wherein R is 【Chemical 2】 selected from the group R-G1 consisting of R 1 R consisting of H and F 1 selected from the R-G1 group, Part = A 1 -CR = A 2 - is selected from the group A-G1 consisting of =N-CR=N-, =N-CR=CH- and =CH-CR=N- Part - L 1 = L 2 - is selected from the group L-G1 consisting of -N=N-, -N=CH- and -CH=N-) and / or its tautomer or a salt thereof.

2. R is [Chemical Formula 3] selected from the group R-G2 consisting of The compound according to claim 1 and / or its tautomer or a salt thereof.

3. R is [Chemical Formula 4] selected from the group R-G3 consisting of The compound according to claim 1 and / or its tautomer or a salt thereof.

4. R is 【Chemical Formula 5】 selected from the group R-G4 consisting of The compound according to claim 1 and / or its tautomer or a salt thereof.

5. Part = A 1 -CR = A 2 -wherein is selected from the group A-G2 consisting of =N-CR=N- The compound according to any one of claims 1 to 4 and / or its tautomer or a salt thereof.

6. Part = A 1 -CR = A 2 -wherein is selected from the group A-G3 consisting of =N-CR=CH- The compound according to any one of claims 1 to 4 and / or its tautomer or a salt thereof.

7. Part = A 1 -CR = A 2 -wherein is selected from the group A-G4 consisting of =CH-CR=N- The compound according to any one of claims 1 to 4 and / or its tautomer or a salt thereof.

8. The compound of formula (I) is 【Chemical Formula 6】 【Chem.】 selected from the group consisting of The compound according to any one of claims 1 to 7 and / or its tautomer or a salt thereof.

9. A pharmaceutically acceptable salt of the compound according to any one of claims 1 to 8 and / or its tautomer.

10. A pharmaceutical composition comprising the compound according to any one of claims 1 to 9 and / or its tautomer, or a pharmaceutically acceptable salt thereof, and optionally one or more inert carriers and / or diluents.

11. The pharmaceutical composition according to claim 10 for the treatment of eye diseases.

12. The pharmaceutical composition according to claim 10 for the treatment of diabetic macular edema, age-related macular degeneration and / or choroidal neovascularization.

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