WO2023110700A1 - Dérivés de dihydrofuropyridine utilisés comme inhibiteurs de la rho-kinase - Google Patents

Dérivés de dihydrofuropyridine utilisés comme inhibiteurs de la rho-kinase Download PDF

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WO2023110700A1
WO2023110700A1 PCT/EP2022/085286 EP2022085286W WO2023110700A1 WO 2023110700 A1 WO2023110700 A1 WO 2023110700A1 EP 2022085286 W EP2022085286 W EP 2022085286W WO 2023110700 A1 WO2023110700 A1 WO 2023110700A1
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methyl
pyridin
amino
dihydrofuro
benzamide
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PCT/EP2022/085286
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English (en)
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Alessandro ACCETTA
Fabio Rancati
Daniele PALA
Adele Elisa PASQUA
David Edward Clark
Christine Edwards
Prashant Bhimrao Kapadnis
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Chiesi Farmaceutici S.P.A.
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Publication of WO2023110700A1 publication Critical patent/WO2023110700A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/04—Ortho-condensed systems
    • C07D491/044—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
    • C07D491/048—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring the oxygen-containing ring being five-membered
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A61P11/06—Antiasthmatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A61P11/08—Bronchodilators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection

Definitions

  • the present invention relates to novel compounds inhibiting Rho Kinase (hereinafter ROCK Inhibitors); methods of preparing such compounds, pharmaceutical compositions containing them and therapeutic use thereof.
  • ROCK Inhibitors novel compounds inhibiting Rho Kinase
  • the compounds of the invention are inhibitors of the activity or function of the ROCK -I and/or ROCK-II isoforms of the Rho-associated coiled-coil forming protein kinase (ROCK).
  • ROCK Rho-associated coiled-coil forming protein kinase
  • Rho-associated coiled-coil forming protein kinase belongs to the AGC (PKA/PKG/PKC) family of serine-threonine kinases.
  • AGC PKA/PKG/PKC
  • ROCK-I also referred to as pl60 ROCK or ROKp or ROCK1
  • ROCK-II ROKa or ROCK2
  • ROCK-II ROKa or ROCK2
  • ROCK-II and ROCK-I are expressed in many human and rodent tissues including the heart, pancreas, lung, liver, skeletal muscle, kidney and brain (above Riento and Ridley, 2003).
  • ROCK activity is significantly higher in both lung tissues and circulating neutrophils as compared with controls (Duong-Quy S, Bei Y, Liu Z, Dinh-Xuan AT. Role of Rho-kinase and its inhibitors in pulmonary hypertension. Pharmacol Ther. 2013;137(3):352-64).
  • a significant correlation was established between neutrophil ROCK activity and the severity and duration of pulmonary hypertension (Duong-Quy et al., 2013).
  • ROCK reactive oxygen species
  • COPD chronic obstructive pulmonary disease
  • ARDS/ALI acute and chronic pulmonary diseases
  • selective inhibitors have the potential to treat a number of pathological mechanisms in respiratory diseases, such as smooth muscle hyper-reactivity, bronchoconstriction, airway inflammation and airway remodeling, neuromodulation and exacerbations due to respiratory tract viral infection (Fernandes LB, Henry PJ, Goldie RG. Rho kinase as a therapeutic target in the treatment of asthma and chronic obstructive pulmonary disease. Ther Adv Respir Dis. 2007 Oct;l(l):25-33).
  • Rho kinase inhibitor Y-27632 causes bronchodilatation and reduces pulmonary eosinophilia trafficking and airways hyperresponsiveness (Gosens, R.; Schaafsma, D.; Nelemans, S. A.; Halayko, A. J. Rhokinase as a drug target for the treatment of airway hyperresponsiveness in asthma. Mini-Rev. Med. Chem. 2006, 6, 339-348). Pulmonary ROCK activation has been demonstrated in humans with idiopathic pulmonary fibrosis (IPF) and in animal models of this disease.
  • IPF idiopathic pulmonary fibrosis
  • ROCK inhibitors can prevent fibrosis in these models, and more importantly, induce the regression of already established fibrosis, thus indicating ROCK inhibitors as potential powerful pharmacological agents to halt progression of pulmonary fibrosis (Jiang, C.; Huang, H.; Liu, J.; Wang, Y.; Lu, Z.; Xu, Z. Fasudil, a rho-kinase inhibitor, attenuates bleomycin-induced pulmonary fibrosis in mice. Int. J. Mol. Sci. 2012, 13, 8293-8307).
  • Rho Kinase Inhibitors See e.g. W02004/039796 disclosing phenylaminopyrimidine compounds derivatives; W02006/009889 disclosing indazole compound derivatives; W02010/032875 disclosing nicotinamide compounds derivatives; W02009/079008 disclosing pyrazole derivatives; WO2014/118133 disclosing pyrimidine derivatives and, of the same Applicant of the present invention, WO2018/115383 disclosing bicyclic dihydropyrimidine and WO 2018/138293, WO 2019/048479, WO 2019/121223, WO 2019/121233, WO 2019/121406, WO 2019/238628, WO 2020/016129 disclosing tyrosine-amide compounds derivatives and analogues.
  • the compounds disclosed exhibit substantial structural differences from the compounds of the present invention.
  • the present invention relates to novel compounds differing from the structures disclosed in the art at least for a common new core scaffold.
  • the invention relates to compounds that are characterized by the 2,3-dihydrofuro[3,2-c]pyridine moiety, particularly 2,3-dihydrofuro[3,2-c]pyridin-4- amine, particularly preferably N-(3-(((2,3-dihydrofuro[3,2-c]pyridin-4- yl)amino)methyl)phenyl)formamide and 3-(((2,3-dihydrofuro[3,2-c]pyridin-4- yl)amino)methyl)benzamide derivatives, particularly N-linked spirocyclic derivatives of such compounds, which are inhibitors of ROCK-I and ROCK-II isoforms of the Rho- associated coiled-coil forming protein kinase (ROCK) that have therapeutically desirable characteristics, particularly promising in the field of respiratory diseases but not excluding other fields such as that of immune system disorders including Graft-versus- host disease (GVHD), and specifically for some pulmonary diseases including asthma, chronic obstruct
  • the compounds of the invention may be prepared for administration by any route consistent with their pharmacokinetic properties.
  • the compound of the invention are active as inhibitors of ROCK-I and ROCK-II isoforms, they are potent and have advantageously other improved properties such as selectivity and other in vitro properties indicative for a preferred route of administration.
  • the present invention is directed to a class of compounds, acting as inhibitors of the Rho Kinase (ROCK), of formula (I)
  • variables Xi, X2, X3 and X4, p, R, Ri, L, n, R2 and R3, Re and R7, q and Y are as defined in the detailed description of the invention; or pharmaceutically acceptable salts and solvates thereof.
  • the present invention refers to a compound of formula (I) for use as a medicament.
  • the present invention provides the use of a compound of the invention for the manufacture of a medicament.
  • the present invention provides the use of a compound of the invention for the preparation of a medicament for the treatment of any disease associated with ROCK enzyme mechanisms, that is to say characterized by ROCK enzyme aberrant activity and/or wherein an inhibition of activity is desirable and in particular through the selective inhibition of the ROCK enzyme isoforms over other Kinases.
  • the present invention provides a method for prevention and/or treatment of any disease associated with ROCK enzyme mechanisms as above defined, said method comprises administering to a patient in need of such treatment a therapeutically effective amount of a compound of the invention.
  • the compounds of the invention are used alone or combined with other active ingredients and may be administered for the prevention and/or treatment of immune system disorders including Graft-versus-host disease (GVHD), and for pulmonary diseases including asthma, chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis (IPF) and pulmonary hypertension (PH) and specifically pulmonary arterial hypertension (PAH).
  • GVHD Graft-versus-host disease
  • COPD chronic obstructive pulmonary disease
  • IPF idiopathic pulmonary fibrosis
  • PH pulmonary hypertension
  • PAH specifically pulmonary arterial hypertension
  • “Pharmaceutically acceptable salts” refers to derivatives of compounds of formula (I) wherein the parent compound is suitably modified by converting any of the free acid or basic group, if present, into the corresponding addition salt with any base or acid conventionally intended as being pharmaceutically acceptable.
  • Suitable examples of said salts may thus include mineral or organic acid addition salts of basic residues such as amino groups, as well as mineral or organic basic addition salts of acid residues such as carboxylic groups.
  • Cations of inorganic bases which can be suitably used to prepare salts of the invention comprise ions of alkali or alkaline earth metals such as potassium, sodium, calcium or magnesium.
  • Those obtained by reacting the main compound, functioning as a base, with an inorganic or organic acid to form a salt comprise, for example, salts of hydrochloric, hydrobromic, sulfuric, phosphoric, methane sulfonic, camphor sulfonic, acetic, oxalic, maleic, fumaric, succinic and citric acids.
  • Many organic compounds can form complexes with solvents in which they are reacted or from which they are precipitated or crystallized. These complexes are known as “solvates” which are a further object of the invention.
  • Halogen or “halogen atoms” includes fluorine, chlorine, bromine, and iodine atom ; meaning Fluoro, Chloro, Bromo, Iodo as substituent.
  • (C 1 -C 6 )Alkyl refers to straight-chained or branched alkyl groups wherein the number of carbon atoms is in the range 1 to 6. Particular alkyl groups are for example methyl, ethyl, n-propyl, isopropyl, t-butyl, 3-methylbutyl and the like.
  • (C 1 -C 6 )Haloalkyl refer to the above defined “(C 1 -C 6 )alkyl” groups wherein one or more hydrogen atoms are replaced by one or more halogen atoms, which can be the same or different from each other. Examples include halogenated, poly-halogenated and fully halogenated alkyl groups wherein all of the hydrogen atoms are replaced by halogen atoms, e.g. trifluoromethyl or difluoro methyl groups.
  • (C 1 -C 6 )Hydroxyalkyl and “(C 1 - C 6 )aminoalkyl”refer to the above defined “(C 1 -C 6 )alkyl” groups wherein one or more hydrogen atoms are replaced by one or more hydroxy (OH) or amino group respectively, examples being hydroxymethyl and aminomethyl and the like.
  • the definition of aminoalkyl encompasses alkyl groups (i.e. “(C 1 -C 6 )alkyl” groups) substituted by one or more amino groups (-NR 10 R 9 ).
  • aminoalkyl is a mono-aminoalkyl group such as R 10 R 9 N-(C 1 -C 6 )alkyl.
  • the substituents R 10 and R 9 are defined as R 4 and R 5 in the above detailed description of the invention.
  • Derived expression such as aminoalkoxyl thus refer to the above define aminoalkyl linked to the rest of the molecule from the alkil side via an ether bridge, e.g. with linear representation -O-(CH 2 ) m NR 4 R 5 .
  • (C 3 -C 10 )cycloalkyl likewise “(C 3 -C 8 )cycloalkyl” or “(C 3 - C 6 )cycloalkyl” refers to saturated cyclic hydrocarbon groups containing the indicated number of ring carbon atoms. Examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl, and polycyclic ring systems such as adamantan-yl.
  • Aryl refers to mono, bi- or tri-cyclic carbon ring systems which have 6 to 20, preferably from 6 to 15 ring atoms, wherein at least one ring is aromatic.
  • heteroaryl refers to mono-, bi- or tri-cyclic ring systems with 5 to 20, preferably from 5 to 15 ring atoms, in which at least one ring is aromatic and in which at least one ring atom is a heteroatom (e.g. N, S or O).
  • aryl or heteroaryl monocyclic ring systems include, for instance, phenyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, isothiazolyl, thiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, furanyl radicals and the like.
  • aryl or heteroaryl bicyclic ring systems include naphthalenyl, biphenylenyl, purinyl, pteridinyl, pyrazolopyrimidinyl, benzotriazolyl, benzoimidazole- yl, quinolinyl, isoquinolinyl, indolyl, isoindolyl, indazolyl, benzothiopheneyl, benzodioxinyl, dihydrobenzodioxinyl, indenyl, dihydro-indenyl, dihydrobenzo[1,4]dioxinyl, benzothiazole-2-yl, dihydrobenzodioxepinyl, benzooxazinyl, 1,2,3,4-tetrahydroisoquinoline-6-yl, 4,5,6,7-tetrahydrothiazolo[4,5-c]pyridine, 4,5,6,7- tetrahydrobenz
  • aryl or heteroaryl tricyclic ring systems include fluorenyl radicals as well as benzocondensed derivatives of the aforementioned heteroaryl bicyclic ring systems.
  • Said heterocycloalkyl (i.e. heterocyclic radical or group) is further optionally substituted on the available points in the ring, namely on a carbon atom, or on an heteroatom available for substitution.
  • heterocycloalkyl are represented by: oxetanyl, tetrahydro-furanyl, pyrrolidinyl, imidazolidinyl, thiazolidinyl, piperazinyl, piperidinyl, morpholinyl, thiomorpholinyl, dihydro- or tetrahydro-pyridinyl, tetrahydropyranyl, pyranyl, 2H- or 4H-pyranyl, dihydro- or tetrahydrofuranyl, dihydroisoxazolyl, pyrrolidin-2-one-yl, dihydropyrrolyl, 5-oxopyrrolidin-3-yl, (1R,5S,6R)-3-oxabicyclo[3.
  • Aryl(C 1 -C 6 )alkyl refers to an aryl ring linked to a straight-chained or branched alkyl group wherein the number of constituent carbon atoms is in the range from 1 to 6, e.g. phenylmethyl (i.e. benzyl), phenylethyl or phenylpropyl.
  • Heteroaryl(C 1 -C 6 )alkyl refers to an heteroaryl ring linked to a straight-chained or branched alkyl group wherein the number of constituent carbon atoms is in the range from 1 to 6, e.g. furanylmethyl.
  • alkanoyl refers to HC(O)- or to alkylcarbonyl groups (e.g. (C 1 -C 6 )alkylC(O)-) wherein the group “alkyl” has the meaning above defined. Examples include formyl, acetyl, propanoyl, butanoyl.
  • alkoxy or “(C 1 -C 10 ) alkoxyl”, likewise “(C 1 -C 6 ) alkoxy” or “(C 1 -C 6 ) alkoxyl” etc., refers to a straight or branched hydrocarbon of the indicated number of carbons, linked to the rest of the molecule through an oxygen bridge.
  • (C 1 -C 6 )Alkylthio refers to the above hydrocarbon linked through a sulfur bridge.
  • the derived expression “(C 1 -C 6 )haloalkoxy” or “(C 1 -C 6 )haloalkoxyl” refers to the above defined haloalkyl, linked through an oxygen bridge.
  • An example of (C 1 -C 6 )haloalkoxy is trifluoromethoxy.
  • (C 3 -C 6 )heterocycloalkyl-(C 1 -C 6 )alkyl and “(C 3 -C 6 )cycloalkyl-(C 1 -C 6 )alkyl” refer to the above defined heterocycloalkyl and cycloalkyl groups linked to the rest of the molecule via an alkyl group of the indicated number of carbons, corresponding e.g. to linear formula (C 3 -C 6 )heterocycloalkyl- (CH 2 ) m - or (C 3 -C 6 )cycloalkyl-(CH 2 ) m- for example piperidin-4-yl-methyl, cyclohexylethyl.
  • (C 1 -C 6 )alkoxy-(C 1 -C 6 )alkyl refers to the above defined alkoxy group linked to the rest of the molecule via an alkyl group of the indicated number of carbons, corresponding e.g. to linear formula (C 1 -C 6 )alkoxy-(CH 2 ) m - for example methoxymethyl.
  • (C 1 -C 6 )haloalkoxy (C 1 -C 6 )alkyl refers to the above defined (C 1 -C 6 )haloalkoxy” group linked to the rest of the molecule via an alkyl group of the indicated number of carbons, for example difluoromethoxypropyl.
  • bracketed group is a lateral group, not included into the chain, and brackets are used, when deemed useful, to help disambiguating linear chemical formulas; e.g. the sulfonyl group -SO 2 - might be also represented as–S(O) 2 – to disambiguate e.g. with respect to the sulfinic group –S(O)O–.
  • a numerical index the statement (value) “p is zero” or “p is 0” means that the substituent or group bearing the index p (e.g. (R)p) is absent, that is to say no substituent, other than H when needed, is present.
  • the index is attached to a bridging divalent group (e.g.
  • physiological acceptable anions selected among chloride, bromide, iodide, trifluoroacetate, formate, sulfate, phosphate, methanesulfonate, nitrate, maleate, acetate, citrate, fumarate, tartrate, oxalate, succinate, benzoate, p- toluenesulfonate, pamoate and naphthalene disulfonate may be present.
  • corresponding physiological cation salts may be present as well, for instance including alkaline or alkaline earth metal ions.
  • Compounds of formula (I) when they contain one or more stereogenic center may exist as optical stereoisomers. Where the compounds of the invention have at least one stereogenic center, they may accordingly exist as enantiomers. Where the compounds of the invention possess two or more stereogenic centers, they may additionally exist as diastereoisomers. It is to be understood that all such single enantiomers, diastereoisomers and mixtures thereof in any proportion are encompassed within the scope of the present invention.
  • the absolute configuration (R) or (S) for carbon bearing a stereogenic center is assigned on the basis of Cahn-Ingold-Prelog nomenclature rules based on groups’ priorities.
  • Single stereoisomer “single diastereoisomer” or “single enantiomer”, when reported near the chemical name of a compound indicate that the isomer was isolated as single diastereoisomer or enantiomer (e.g via chiral chromatography) but the absolute configuration at the relevant stereogenic center was not determined/assigned.
  • Atropisomers result from hindered rotation about single bonds where the steric strain barrier to rotation is high enough to allow for the isolation of the conformers (Bringmann G et al, Angew. Chemie Int. Ed. 44 (34), 5384-5427, 2005. doi : 10.1002/anie.200462661 ).
  • Atropisomers differ from other chiral compounds in that in many cases they can be equilibrated thermally whereas in the other forms of chirality isomerization is usually only possible chemically.
  • Atropisomers Separation of atropisomers is possible by chiral resolution methods such as selective crystallization.
  • one atropisomer is formed at the expense of the other.
  • Atroposelective synthesis may be carried out by use of chiral auxiliaries like a Corey Bakshi Shibata (CBS) catalyst, an asymmetric catalyst derived from proline, or by approaches based on thermodynamic equilibration when an isomerization reaction favors one atropisomer over the other.
  • CBS Corey Bakshi Shibata
  • Racemic forms of compounds of formula (I) as well as the individual atropisomers (substantially free of its corresponding enantiomer) and stereoisomer-enriched atropisomer mixtures are included in the scope of the present invention.
  • deuterated derivative means that at least one position occupied by a hydrogen atom is occupied by deuterium in an amount above its natural abundance.
  • the percent of deuterium at that position is at least 90%, more preferably at least 95%, even more preferably 99%.
  • the present invention is directed to a class of compounds of formula (I) I wherein X 1 , X 2 , X 3 and X 4 are all CH or one of X 1 , X 2 , X 3 and X 4 is N and the others are CH; preferably X 1 , X 2 , X 3 and X 4 are all CH; Y is selected from N, or O; q is in each occurrence independently selected from 1 or 2; p is zero or an integer from 1 to 4; each R, when present, is in each occurrence independently selected from (C 1 -C 6 )alkyl and halogen selected from F, Cl, Br and I; wherein preferably R is selected from F, Cl and methyl; R 1 is pyrazolyl, preferably1H-pyrazol-4-yl or pyridinyl, preferably pyridin-4-yl or pyrimidinyl, preferably pyrimidin-4yl, substituted by one or more group selected from - (I) I where
  • the invention is directed to a compound of formula (I) wherein X 1 , X 2 , X 3 and X 4 are all CH; Y is -N-; q is in each occurrence independently selected from 1 or 2; p is zero or an integer from 1 to 4; each R, when present, is an halogen in each occurrence independently selected from F, Cl, Br and I, wherein preferably R is F; R 1 is pyrimidinyl substituted by -NH 2 ; particularly preferably R 1 is 2- aminopyrimidin-4-yl; L is -C(O)NH-; n is 0; R 3 is absent and R 2 is selected from the group consisting of-H, (C 1 -C 6 ) alkoxy (C 1 -C 6 )alkyl, preferably (C 1 -C 6 )alkoxy-(CH 2 ) m -, ciano (C1-C6)alkyl, meaning NC-(C1-C6)alkyl, (
  • a further preferred group of compounds according to the invention are those of formula (I) wherein X 1 , X 2 , X 3 and X 4 are all CH; p is zero or 1; each R, when present, is F; R 1 is selected from1H-pyrazol-4-yl, pyridin-4-yl and 2-aminopyrimidin-4-yl; L is -C(O)NH-; n is in each occurrence independently 0 or an integer selected from 1, 2 or 3; R 3 , is H, and R 2 is absent when Y is O, or when Y is N, R 2 is selected from the group consisting of -H, (C 1 -C 6 ) alkoxy (C 1 -C 6 )alkyl, which is 2-methoxyethyl, ciano (C 1 -C 6 )alkylwhich is 2-cyanoethyl, (C 1 -C 6 ) haloalkyl,which is 2-fluor
  • the invention also provides a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof in admixture with one or more pharmaceutically acceptable carriers or excipients, either alone or in combination with one or more further active ingredient as detailed below.
  • the invention provides the compounds listed in the table below and pharmaceutical acceptable salts thereof.
  • the compounds of the invention can be prepared from readily available starting materials using the following general methods and procedures or by using slightly modified processes readily available to those of ordinary skill in the art. Although a particular embodiment of the present invention may be shown or described herein, those skilled in the art will recognize that all embodiments or aspects of the present invention can be prepared using the methods described herein or by using other known methods, reagents and starting materials. When typical or preferred process conditions (i.e. reaction temperatures, times, mole ratios of reactants, solvents, pressures, etc.) are given, other process conditions can also be used unless otherwise stated. While the optimum reaction conditions may vary depending on the particular reactants or solvent used, such conditions can be readily determined by those skilled in the art by routine optimization procedures.
  • process conditions i.e. reaction temperatures, times, mole ratios of reactants, solvents, pressures, etc.
  • PG protective groups
  • a suitable protective group for intermediates requiring protection of a carboxylic acid can be C1-C4 esters (PGi: methyl, isopropyl, tert-butyl or ethyl), preferably methyl.
  • a suitable protective group for protecting NH of R1 groups can be tert-butoxycarbonyl (Boc) or a tetrahydro-2H-pyran-2-yl (THP).
  • a suitable protective group for intermediates requiring the amino group protection can be carbamates such as tert-butylcarbamate (tertbutoxycarbonyl or Boc), benzylcarbamate (Benzyloxycarbonyl or Cbz), ethylcarbamate (ethoxycarbonyl) or methylcarbamate (methoxycarbonyl), preferably PG3 is Boc.
  • Compounds of formula (I) can contain one or more stereogenic centre.
  • Enantiomerically pure compounds can be prepared according to generally known reactions, e.g. according to the reactions described below, by means of enantiomerically pure starting materials and intermediates. These intermediates may be commercially available or readily produced from commercial sources by those of ordinary skill in the art.
  • enantiomerically pure compounds can be prepared from the corresponding racemates by means of chiral chromatography purification.
  • Stereochemically pure compounds may be obtained by chiral separation from a stereoisomers mixture, or (whenever there are two or more stereogenic centres -i.e. chiral center- in compounds of formula (I)) stepwise by chromatographic separation of diastereoisomers followed by further chiral separation into single stereoisomers.
  • Intermediate II can be converted into intermediate III by means of four consecutive steps including 1) chlorination, 2) amination, 3) reduction and 4) bromination.
  • the chlorination step may be carried out by refluxing intermediate II with an appropriate chlorinating agent (neat or in solution with an organic solvent such as DCM or dioxane) such as POCE or SOCh.
  • an appropriate chlorinating agent nitrogen or in solution with an organic solvent such as DCM or dioxane
  • POCE or SOCh organic solvent
  • the amination step can be carried out by introducing a masked ammonia such as benzophenone imine through a Buchwald type palladium catalyzed reaction using, for example, tris(dibenzylideneacetone)dipalladium(0)/BINAP catalytic system followed by hydrolysis of the benzophenone imine by using hydroxylamine to give the corresponding furo[3,2-c]pyridin-4-amine.
  • the amination step can be carried out by introducing 4-methoxybenzylamine by means of SNAT reaction (nucleophilic aromatic substitution) followed by deprotection with a strong acid such as trifluoroacetic acid or methanesulfonic acid.
  • Reduction of furo[3,2-c]pyridin-4-amine to give 2,3- dihydrofuro[3,2-c]pyridin-4-amine can be carried out, for example, by hydrogenation of a solution of furo[3,2-c]pyridin-4-amine in methanol / acetic acid in the presence of a Pd/C catalyst under high H2 pressure (e.g. 10 bar) and at a temperature of 50°C or higher.
  • intermediate III can be obtained by means of bromination of 2,3- dihydrofuro[3,2-c]pyridin-4-amine (step 4) by reaction with a brominating agent such as N-bromosuccinimide in a polar aprotic solvent such as acetonitrile or tetrahydrofuran for a few hours at low temperature (e.g. -10 - 0 °C).
  • a brominating agent such as N-bromosuccinimide
  • a polar aprotic solvent such as acetonitrile or tetrahydrofuran
  • Intermediate III and carbonyl intermediate IV can be combined to give intermediate V through a reductive amination reaction that can be performed in an appropriate solvent such as DCM or THF, in the presence of a Lewis acid such as chloro(triisopropoxy)titanium(IV) or titanium tetraisopropoxide(IV) followed by addition of a reducing agent such as sodium triacetoxyborohydride or sodium cyanoborohydride, in the presence of an organic acid such as acetic acid or trifluoroacetic acid.
  • a Lewis acid such as chloro(triisopropoxy)titanium(IV) or titanium tetraisopropoxide(IV)
  • a reducing agent such as sodium triacetoxyborohydride or sodium cyanoborohydride
  • Intermediate V can be converted into intermediate VI by a direct introduction of group Ri through a metal/palladium catalyzed cross coupling reaction such as Stille coupling, Suzuki coupling or similar (Strategic application of named reactions in organic synthesis, L. Kurti, B. Czako, Ed. 2005).
  • a metal/palladium catalyzed cross coupling reaction such as Stille coupling, Suzuki coupling or similar (Strategic application of named reactions in organic synthesis, L. Kurti, B. Czako, Ed. 2005).
  • a suitable palladium catalyzed cross coupling for introducing Ri when it is an 2-aminopyrimidin-4-yl is a Stille coupling.
  • a Stille coupling can be performed by reacting intermediate V with the corresponding organostannane of group Ri, in the presence of a Pd catalyst such as tetrakistriphenylphosphinepalladium(O), tris(dibenzylideneacetone)dipalladium(0), or PdC12(dppf)2, in an organic solvent such as dioxane or THF or DMF, in the presence of a copper(I) salt such as copper(I) thiophene- 2-carboxylate, under heating (90-150°C).
  • a Pd catalyst such as tetrakistriphenylphosphinepalladium(O), tris(dibenzylideneacetone)dipalladium(0), or PdC12(dppf)2
  • an organic solvent such as dio
  • Organostannanes are generally commercially available or may be readily prepared by those skilled in the art starting from corresponding commercially available halides. Experimental procedures for the preparation of an organostannane not commercially available are reported in the experimental section.
  • Ri is a 2-aminopyrimidin-4-yl, for synthetic convenience, the amino group needs to be masked/protected during the Stille coupling. Said amino group may be suitably protected (PG2) by one or even two Boc groups and removed when convenient trougthout the synthetic sequence.
  • Suzuki coupling can be performed by reacting intermediate V with the corresponding boronic acid or boron pinacolate ester of group Ri, in the presence of a Pd catalyst such as tris(dibenzylideneacetone)dipalladium(0), PdC12(dppf)2.DCM adduct or tetrakistriphenylphosphinepalladium(O), in an organic solvent such as dioxane, THF or DMF with or without water, with an inorganic base such as an alkaline carbonate (for example CS2CO3) or an inorganic phosphate (for example K3PO4), under heating (90- 150°C).
  • a Pd catalyst such as tris(dibenzylideneacetone)dipalladium(0), PdC12(dppf)2.DCM adduct or tetrakistriphenylphosphinepalladium(O)
  • organic solvent such as dioxane, THF or DMF with or without
  • Boronic acid and boronic pinacolate esters are generally commercially available or may be readily prepared by those skilled in the art starting from commercially available reagents.
  • Ri is pyrazolyl
  • the NH needs to be masked/protected during the Suzuki coupling.
  • Said NH may be suitably protected (PG2) by THP or Boc and removed when convenient througthout the synthetic sequence.
  • intermediate VI can be obtained by inverting the order of reductive amination and Palladium cross coupling reaction.
  • intermediate III can be converted into intermediate IX by Palladium catalyzed cross coupling in the same way as described above for conversion of intermediate V in VI, then intermediate IX can be converted by a reductive amination reaction into intermediate VI in the same way as described above for conversion of intermediate III into V.
  • Removal of PGi (when PGi is methyl) from intermediate VI to give the intermediate VII may be carried out by hydrolysis, using an inorganic base such as LiOH or NaOH in a mixture of an organic solvent such as THF and/or methanol with water, generally at RT and for a time ranging from 1 h to overnight.
  • an inorganic base such as LiOH or NaOH
  • a mixture of an organic solvent such as THF and/or methanol with water
  • Removal of PG2 (when PG2 is Boc) from intermediate VI to give the intermediate VII may be carried out by acidic deprotection.
  • an acidic Boc cleavage may be carried out by means of concentrated hydrochloric acid or trifluoroacetic acid.
  • Boc groups on bis-Boc protected 2-aminopyrimidin-4-yl can also be cleaved.
  • RI is a pyrazolyl group protected by THP
  • said protective group can be easily removed by heating (up to 100°C) the protected precursor with concentrated aqueous acid such as 15 to 30% w/w aqueous hydrochloric acid or up to 30%w/w aqueous sulfuric acid for a time up to 1 h or less.
  • concentrated aqueous acid such as 15 to 30% w/w aqueous hydrochloric acid or up to 30%w/w aqueous sulfuric acid for a time up to 1 h or less.
  • PGI when it is Me can be removed concomitantly.
  • Reaction between acid intermediate VII and amino intermediate Villa to give a compound of formula (I) may be carried out under suitable amide coupling reaction conditions.
  • acid intermediate VII may be reacted in the presence of an activating agent such as TBTU, HATU or COMU, in the presence of an organic base such as DIPEA or TEA, in a suitable organic solvent such as DCM or DMF, and at a temperature generally around RT for a time ranging from a few hours to overnight.
  • PG3 generally Boc
  • a compound of formula (la) wherein R2 is in each occurrence different from H such compounds can be obtained by further elaboration of a compound of formula (la) (wherein R2 is H) by means of an alkylation reaction, a reductive amination reaction or an amidation reaction by using generally known methods (Scheme 2).
  • an alkylation can be carried out by reaction of the amine (compound of formula I wherein Y is N and R2 is H) with an alkylating agent such as a alkylbromide, iodide, tosylate or similar reagents, in an organic solvent such as DMF or DCM, in the presence of an inorganic base such as K2CO3 at room temperature or higher.
  • reaction can be carried out by reacting said amine with the a suitable aldehyde or ketone in an organic solvent such as DMF, DCM or similar, in the presence of an acid such as acetic acid and a suitable reducing agent such as sodium triacetoxyborohydride, sodium cyanoborohydride or similar well known methods to those skilled in the art.
  • a suitable aldehyde or ketone in an organic solvent such as DMF, DCM or similar
  • an acid such as acetic acid
  • a suitable reducing agent such as sodium triacetoxyborohydride, sodium cyanoborohydride or similar well known methods to those skilled in the art.
  • Amidation on a compound of formula (la) (wherein R2 is H) can be carried out by reaction of the said amine with the a suitable carboxylic acid in the presence of a coupling agent such as HATU, TBTU or COMU, in an organic solvent such as DMF and/or DCM and in the presence of an organic base such as DIPEA.
  • a coupling agent such as HATU, TBTU or COMU
  • organic solvent such as DMF and/or DCM
  • DIPEA organic base
  • compounds of the invention are inhibitors of kinase activity, in particular Rho-kinase activity.
  • the invention provides a compound of formula (I) for use as a medicament, preferably for the prevention and /or treatment of pulmonary disease.
  • the invention provides the use of a compound (I), or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of disorders associated with ROCK enzyme mechanisms, including immune system disorders and particularly for the treatment of disorders such as pulmonary diseases.
  • the invention provides compounds of formula (I) for use in the prevention and /or treatment of immune system disorders including Graft-versus-host disease (GVHD), and for pulmonary disease selected from the group consisting of asthma, chronic obstructive pulmonary disease COPD, idiopathic pulmonary fibrosis (IPF), pulmonary hypertension (PH) and specifically Pulmonary Arterial Hypertension (PAH).
  • GVHD Graft-versus-host disease
  • pulmonary disease selected from the group consisting of asthma, chronic obstructive pulmonary disease COPD, idiopathic pulmonary fibrosis (IPF), pulmonary hypertension (PH) and specifically Pulmonary Arterial Hypertension (PAH).
  • the invention provides a method for the prevention and/or treatment of disorders associated with ROCK enzymes mechanisms, said method comprising administering to a patient in need of such treatment a therapeutically effective amount of a compound of the invention.
  • the invention provides methods for the prevention and/or treatment wherein the disorder is an immune system disorder such as Graft-versus-host disease (GVHD), and/or a respiratory disease selected from asthma, chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis (IPF), Pulmonary hypertension (PH) and specifically Pulmonary Arterial Hypertension (PAH).
  • GVHD Graft-versus-host disease
  • COPD chronic obstructive pulmonary disease
  • IPF idiopathic pulmonary fibrosis
  • PH Pulmonary hypertension
  • PAH Pulmonary Arterial Hypertension
  • ROCK inhibitors may be useful in the treatment of many disorders associated with ROCK enzyme mechanisms.
  • the disorders that can be treated by the compounds of the present invention include glaucoma, inflammatory bowel disease (IBD), immune system disorders including Graft-versus-host disease (GVHD), and pulmonary diseases selected from asthma, chronic obstructive pulmonary disease (COPD), interstitial lung disease such as idiopathic pulmonary fibrosis (IPF) and pulmonary arterial hypertension (PAH).
  • IBD inflammatory bowel disease
  • GVHD Graft-versus-host disease
  • pulmonary diseases selected from asthma, chronic obstructive pulmonary disease (COPD), interstitial lung disease such as idiopathic pulmonary fibrosis (IPF) and pulmonary arterial hypertension (PAH).
  • the disorder that can be treated by the compound of the present invention is selected from the group consisting of asthma, chronic obstructive pulmonary disease (COPD) and interstitial lung disease such as idiopathic pulmonary fibrosis (IPF) and pulmonary arterial hypertension (PAH).
  • COPD chronic obstructive pulmonary disease
  • IPF interstitial lung disease
  • PAH pulmonary arterial hypertension
  • the disorder is selected from asthma, chronic obstructive pulmonary disease (COPD).
  • COPD chronic obstructive pulmonary disease
  • the methods of treatment of the invention comprise administering a safe and effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof to a patient in need thereof.
  • safe and effective amount in reference to a compound of formula (I) or a pharmaceutically acceptable salt thereof or other pharmaceutically-active agent means an amount of the compound sufficient to treat the patient's condition but low enough to avoid serious side effects and it can nevertheless be routinely determined by the skilled artisan.
  • the compounds of formula (I) or pharmaceutically acceptable salts thereof may be administered once or according to a dosing regimen wherein a number of doses are administered at varying intervals of time for a given period of time. Typical daily dosages may vary depending upon the particular route of administration chosen.
  • the invention also provides pharmaceutical compositions of compounds of formula (I) in admixture with one or more pharmaceutically acceptable carrier or excipient, for example those described in Remington’s Pharmaceutical Sciences Handbook, XVII Ed., Mack Pub. , N.Y., U.S.A.
  • the present invention is also directed to use of the compounds of the invention and their pharmaceutical compositions for various route of administration
  • Administration of the compounds of the invention and their pharmaceutical compositions may be accomplished according to patient needs, for example, orally, nasally, parenterally (subcutaneously, intravenously, intramuscularly, intrastemally and by infusion), by inhalation, rectally, vaginally, topically, locally, transdermally, and by ocular administration.
  • solid oral dosage forms can be used for administering compounds of the invention including such solid forms as tablets, gelcaps, capsules, caplets, granules, lozenges and bulk powders.
  • the compounds of the present invention can be administered alone or combined with various pharmaceutically acceptable carriers, diluents (such as sucrose, mannitol, lactose, starches) and known excipients, including suspending agents, solubilizers, buffering agents, binders, disintegrants, preservatives, colorants, flavorants, lubricants and the like.
  • diluents such as sucrose, mannitol, lactose, starches
  • excipients including suspending agents, solubilizers, buffering agents, binders, disintegrants, preservatives, colorants, flavorants, lubricants and the like.
  • Time release capsules, tablets and gels are also advantageous.
  • liquid oral dosage forms can also be used for administering compounds of the invention, including aqueous and non-aqueous solutions, emulsions, suspensions, syrups, and elixirs.
  • dosage forms can also contain suitable known inert diluents such as water and suitable known excipients such as preservatives, wetting agents, sweeteners, flavorants, as well as agents for emulsifying and/or suspending the compounds of the invention.
  • the compounds of the present invention may be formulated as injectable composition, for example to be injected intravenously, in the form of an isotonic sterile solution. Other preparations are also possible.
  • Suppositories for rectal administration of the compounds of the invention can be prepared by mixing the compound with a suitable excipient such as cocoa butter, salicylates and polyethylene glycols.
  • Formulations for vaginal administration can be in the form of cream, gel, paste, foam, or spray formula containing, in addition to the active ingredient, such as suitable carriers, are also known.
  • the pharmaceutical composition can be in the form of creams, ointments, liniments, lotions, emulsions, suspensions, gels, solutions, pastes, powders, sprays, and drops suitable for administration to the skin, eye, ear or nose. Topical administration may also involve transdermal administration via means such as transdermal patches.
  • Some preferred compounds of the invention exhibit profile suitable for inhalatory route administration.
  • Drugs optimized for inhaled delivery require certain characteristics that allow the compound, when administered to the lung to maintain a sufficient local concentration (lung retention) to exert a pharmacological effect of the desired duration, and non-relevant levels in unwanted compartments (i.e. plasma).
  • lung retention a sufficient local concentration
  • a pharmacological effect of the desired duration i.e. plasma
  • non-relevant levels in unwanted compartments i.e. plasma
  • one or more features of a compounds need to be optimized such as, and not limited to, membrane permeability, dissolution rate and the degree of basicity.
  • permeability is low
  • dissolution rate is sufficiently slow
  • a basic group is present to enhance binding to the phospholipid-rich lung tissue or toallow lysosomial trapping.
  • compounds of the invention show one or more of the features above that are desirable for an inhaled compound.
  • Other preferred compounds of the invention exhibit a profile suitable for the oral route of administration.
  • Drugs optimized for oral delivery require certain characteristics that allow the orally administered compound to be absorbed by the GI (gastrointestinal) tract and to be poorly cleared in order to give a good bioavailability (F%), thus to maintain a sufficient concentration in plasma and target tissues for a time adequate to sustain pharmacological effect.
  • one or more features of the compounds need to be optimized such as, and not limited to, membrane permeabilty and in vivo clearance. In this respect, to attain high oral bioavailability membrane permeability is high and compounds have reduced metabolic hot spots to (optimized in- vitro clearance).
  • compounds of the invention show one or more of the features above for an oral compound.
  • the compounds according to the invention may be administered by inhalation.
  • Inhalable preparations include inhalable powders, propellant-containing metering aerosols or propellant-free inhalable formulations.
  • the powder may be filled in gelatine, plastic or other capsules, cartridges or blister packs or in a reservoir.
  • a diluent or carrier usually non-toxic and chemically inert to the compounds of the invention, e.g. lactose or any other additive suitable for improving the respirable fraction may be added to the powdered compounds of the invention.
  • Inhalation aerosols containing propellant gas such as hydrofluoroalkanes may contain the compounds of the invention either in solution or in dispersed form.
  • the propellant-driven formulations may also contain other ingredients such as co-solvents, stabilizers and optionally other excipients.
  • the propellant-free inhalable formulations comprising the compounds of the invention may be in the form of solutions or suspensions in an aqueous, alcoholic or hydroalcoholic medium and they may be delivered by jet or ultrasonic nebulizers known from the prior art or by soft-mist nebulizers such as Respimat®.
  • the invention provides compounds of formula (I) and/or pharmaceutical compositions thereof, for use via inhalatory route of administration particularly in the prevention and /or treatment of asthma, chronic obstructive pulmonary disease COPD, idiopathic pulmonary fibrosis (IPF), pulmonary hypertension (PH) and specifically Pulmonary Arterial Hypertension (PAH); preferably in the prevention and /or treatment of asthma, chronic obstructive pulmonary disease COPD.
  • inhalatory route of administration particularly in the prevention and /or treatment of asthma, chronic obstructive pulmonary disease COPD, idiopathic pulmonary fibrosis (IPF), pulmonary hypertension (PH) and specifically Pulmonary Arterial Hypertension (PAH); preferably in the prevention and /or treatment of asthma, chronic obstructive pulmonary disease COPD.
  • the invention provides compounds of formula (I) and/or pharmaceutical compositions thereof, for use via oral route of administration particularly in the prevention and /or treatment of asthma, chronic obstructive pulmonary disease COPD, idiopathic pulmonary fibrosis (IPF), pulmonary hypertension (PH) and specifically Pulmonary Arterial Hypertension (PAH), preferably in the prevention and /or treatment of pulmonary hypertension (PH) and specifically Pulmonary Arterial Hypertension (PAH).
  • COPD chronic obstructive pulmonary disease
  • IPF idiopathic pulmonary fibrosis
  • PH pulmonary hypertension
  • PAH Pulmonary Arterial Hypertension
  • PAH Pulmonary Arterial Hypertension
  • the compounds of the invention can be administered as the sole active agent or in combination (i.e. as co-therapeutic agents administered in fixed dose combination or in combined therapy of separately formulated active ingredients) with other pharmaceutical active ingredients selected from organic nitrates and NO donors; inhaled NO; stimulator of soluble guanylate cyclase (sGC); prostaciclin analogue PGI2 and agonist of prostacyclin receptors; compounds that inhibit the degradation of cyclic guanosine monophosphate (cGMP) and/or cyclic adenosine monophosphate (cAMP), such as inhibitors of phosphodiesterases (PDE) 1, 2, 3, 4 and/or 5, especially PDE 5 inhibitors; human neutrophilic elastase inhibitors; compounds inhibiting the signal transduction cascade, such as tyrosine kinase and/or serine/threonine kinase inhibitors; antithrombotic agents, for example platelet aggregation inhibitors, anticoagulants
  • the compounds of the invention are dosed in combination with phosphodiesterase V such as sildenafil, vardenafil and tadalafil; organic nitrates and NO donors (for example sodium nitroprusside, nitroglycerin, isosorbide mononitrate, isosorbide dinitrate, molsidomine or SIN-1 , and inhaled NO); synthetic prostacyclin analogue PGI2 such as iloprost, treprostinil, epoprostenol and beraprost; agonist of prostacyclin receptors such as selexipag and compounds of WO 2012/007539; stimulators of soluble guanylate cyclase (sGC) like riociguat and tyrosine kinase like imatinib, sorafenib and nilotinib and endothelin antagonist (for example macitentan, bosentan, sitax
  • the compounds of the invention are dosed in combination with beta2 -agonists such as salbutamol, salmeterol, and vilanterol, corticosteroids such as fluticasone propionate or furoate, flunisolide, mometasone furoate, rofleponide and ciclesonide, dexametasone, anticholinergic or antimuscarinic agents such as ipratropium bromide, oxytropium bromide, tiotropium bromide, oxybutynin, and combinations thereof.
  • beta2 -agonists such as salbutamol, salmeterol, and vilanterol
  • corticosteroids such as fluticasone propionate or furoate, flunisolide, mometasone furoate, rofleponide and ciclesonide
  • dexametasone anticholinergic or antimuscarinic agents
  • anticholinergic or antimuscarinic agents such as ipratropium
  • the compounds of the invention are dosed in combination with mitogen-activated protein kinases (P38 MAP kinase) inhibitors, nuclear factor kappa-B kinase subunit beta (IKK2) inhibitors, leukotriene modulators, non-steroidal anti-inflammatory agents (NSAIDs), mucus regulators, mucolytics, expectorant/mucokinetic modulators, peptide mucolytics inhibitors of JAK, SYK inhibitors, inhibitors of PI3Kdelta or PI3Kgamma.
  • mitogen-activated protein kinases P38 MAP kinase
  • IKK2 nuclear factor kappa-B kinase subunit beta
  • NSAIDs non-steroidal anti-inflammatory agents
  • mucus regulators mucolytics
  • expectorant/mucokinetic modulators peptide mucolytics inhibitors of JAK, SYK inhibitors, inhibitors of PI3Kdelta or PI3Kgamma.
  • the invention is also directed to a kit comprising the pharmaceutical compositions of compounds of the invention alone or in combination with or in admixture with one or more pharmaceutically acceptable carriers and/or excipients and a device which may be a single- or multi-dose dry powder inhaler, a metered dose inhaler or a nebulizer.
  • the dosages of the compounds of the invention depend upon a variety of factors including the particular disease to be treated, the severity of the symptoms, the route of administration, the frequency of the dosage interval, the particular compound utilized, the efficacy, toxicology profile, and pharmacokinetic profile of the compound.
  • the compounds of formula (I) can be administered for example, at a dosage comprised between 0.001 and 10000 mg/day, preferably between 0.1 and 500 mg/day.
  • the compounds of formula (I) are administered by inhalation route, they are preferably given at a dosage comprised between 0.001 and 500 mg/day, preferably between 0.1 and 100 mg/day.
  • a pharmaceutical composition comprising a compound of the invention suitable to be administered by inhalation is in various respirable forms, such as inhalable powders (DPI), propellant-containing metering aerosols (PMDI) or propellant-free inhalable formulations (e.g. UDV).
  • DPI inhalable powders
  • PMDI propellant-containing metering aerosols
  • UDV propellant-free inhalable formulations
  • the invention is also directed to a device comprising the pharmaceutical composition comprising a compound according to the invention, which may be a single- or multi-dose dry powder inhaler, a metered dose inhaler and a nebulizer particularly soft mist nebulizer.
  • a compound according to the invention which may be a single- or multi-dose dry powder inhaler, a metered dose inhaler and a nebulizer particularly soft mist nebulizer.
  • the compounds according to the invention can be administered by inhalation; they may be in some case preferably be administered by the oral route.
  • the compounds of formula (I) are administered by oral route, they are preferably given at a dosage comprised from 0.001 mg to 100 mg per kg body weight of a human, often 0.01 mg to about 50 mg per kg, for example 0.1 to 10 mg per kg, in single or multiple doses per day.
  • a pharmaceutical composition comprising a compound of the invention suitable to be administered by the oral route can be in various solid or liquid forms, such as tablets, gelcaps, capsules, caplets, granules, lozenges and bulk powders or aqueous and nonaqueous solutions, emulsions, suspensions, syrups, and elixirs formulations.
  • Isolute® SCX-2 cartridge refers to a pre-packed polypropylene column containing a non-end-capped propylsulfonic acid functionalised silica strong cation exchange sorbent.
  • NMR spectra were obtained on a Bruker Avance 400 MHz, 5mm QNP probe H, C, F, P, single Z gradient, two channel instrument running TopSpin 2.1, or on a Bruker Avance III 400 MHz, 5mm BBFO Plus probe, single Z gradient, two channel instrument running TopSpin 3.0, or on a Varian Unity Inova 400 spectrometer with a 5 mm inverse detection triple resonance probe operating at 400 MHz. Chemical shift are reported as 6 values in ppm relative to tetramethylsilane.
  • ACN acetonitrile
  • BINAP 2,2'-Bis(diphenylphosphino)-l,l'-binaphthalene
  • COMU ((l-Cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholino- carbenium hexafluorophosphate), dba (Dibenzylideneacetone), DCM (dichloromethane), DIPEA or DIEA (N-Ethyldiisopropylamine), DMF (N,N-Dimethylformamide), DMSO (Dimethylsulfoxide), dppf (1,1′-Ferrocenediyl-bis(diphenylphosphine)), EtOH (ethanol), EtOAc (ethyl acetate), FA (Formic acid), HATU (1-[Bis(dimethylamino)methylene]- 1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate, N-[(Dimethylamino)- 1H-1,2,3-triazolo
  • the reaction was quenched with aqueous saturated NaHCO 3 (500 mL), water (500 mL), EtOAc (1000 mL) and aqueous 5% NaCl (500 mL).
  • the resulting organic and aqueous phases were separated, and the aqueous layer further washed with EtOAc (1000 mL).
  • the combined organic layers were washed with aqueous 5% NaCl (7 x 2000 mL) and concentrated to dryness.
  • the residual solid was treated with a mixture of EtOAc (500 mL) and water (200 mL), placed in a sonic bath for some minutes and acidified with aqueous 10% KHSO 4 (300 mL). The solid that appeared was collected by filtration.
  • the biphasic filtrate was partitioned, and the organic layer washed twice with aqueous 10% KHSO 4 (200 mL each). The combined aqueous layer was washed with EtOAc (3 x 500 mL) and mixed with the previous collected solid. The resulting aqueous mixture was neutralized to pH7 with NaHCO 3 and extracted with EtOAc (3 x 1000 mL). The combined organic phase was washed with saturated aqueous NaCl (500 mL), dried over anhydrous MgSO 4 , and concentrated to give the title compound as a solid (27.1 g).
  • Step b 2-(2-Fluoroethyl)-2-azaspiro[3.3]heptan-6-amine; trifluoroacetic acid salt
  • a solution of Intermediate 6a 42 mg, 0.163 mmol
  • dry DCM 5 mL
  • trifluoroacetic acid 0.19 mL, 2.44 mmol
  • Example 1 3-(((7-(2-Aminopyrimidin-4-yl)-2,3-dihydrofuro[3,2-c]pyridin-4- yl)amino)methyl)-N-(2-(2-fluoroethyl)-2-azaspiro[3.3]heptan-6-yl)benzamide (Example 1) To a suspension of Intermediate 2 (59 mg, 0.163 mmol) and Intermediate 6 (44 mg, 0.163 mmol) in DMF (3 mL) was added DIPEA (0.085 mL, 0.488 mmol) and then TBTU (57 mg, 0.179 mmol). The reaction mixture was stirred at room temperature for 18 h.
  • reaction mixture was concentrated and the residue purified by MDAP (Xbridge Phenyl 3x50mm, 3um 5-95% MeOH / H2O (10mM NH 4 CO 3 ), 1.7ml/min, RT) then further purified by reverse phase chromatography onC18 cartridge eluting with water/acetonitrile (+ 0.1% NH 4 OH) 10-95%). The relevant fractions were dried to give the title compound (20 mg).
  • Example 4 3-(((7-(2-aminopyrimidin-4-yl)-2,3-dihydrofuro[3,2-c]pyridin-4- yl)amino)methyl)-N-(7-azaspiro[3.5]nonan-2-yl)benzamide (Example 4)
  • TFA 1.3 mL, 16.8 mmol
  • the solution was diluted with methanol then applied to a 5 g SCX-2 cartridge, which was washed with methanol then eluted using 2N ammonia in methanol.
  • Example 5 3-(((7-(pyridin-4-yl)-2,3-dihydrofuro[3,2-c]pyridin-4-yl)amino)methyl)-N-(7- azaspiro[3.5]nonan-2-yl)benzamide (Example 5)
  • the example 5 was prepared in a similar way of example 4 by replacing intermediate 7a with intermediate 7i.
  • Example 16 N-(7-Acetyl-7-azaspiro[3.5]nonan-2-yl)-3-(((7-(2-aminopyrimidin-4-yl)-2.,3- dihydrofuro[3.,2-c]pyridin-4-yl)amino)methyl)benzamide (Example 16) To a stirring suspension of Example 4 (51 mg, 0.105 mmol), acetic acid (0.0072 mL, 0.126 mmol) and DIPEA (0.055 mL, 0.315 mmol) in DCM (3 mL) and DMF (1 mL) was added HATU (60 mg, 0.158 mmol). The reaction mixture was stirred overnight.
  • reaction mixture was diluted with methanol and applied to a an SCX-2 cartridge, that was washed with methanol then eluted using 2N ammonia in methanol.
  • the basic fraction was evaporated to give a crude product that was purified by MDAP (Xbridge Phenyl 19x150mm, lOum 20-80% MeOH / H 2 O (lOmM NH4CO3), 20mL/min, RT) to give the title compound (28 mg).
  • Example 19 3-(((7-(2-Aminopyrimidin-4-yl)-2,3-dihydrofuro[3,2-c]pyridin-4- yl)amino)methyl)-N-(3-(oxetan-3-yl)-3-azaspiro[5.5]undecan-9-yl)benzamide (Example 19) To a solution of Intermediate 8a (100 mg, 0.195 mmol, 1.00 eq) in DCM (5.00 mL) and DMF (1 mL) was added 3-oxetanone (0.015 mL, 0.234 mmol) and then acetic acid (0.011 mL, 0.195 mmol).
  • reaction mixture was sonicated for 10 minutes to aid dissolution.
  • sodium triacetoxyborohydride (83 mg, 0.389 mmol) was added and the reaction mixture was sonicated for a further 5 minutes and then allowed to stir at RT for 18h.
  • the reaction was quenched with water (1 mL), diluted with DCM and washed with aqueous saturated NaHCO 3 .
  • the organic layer was separated and washed with aqueous saturated NaCl, then dried over magnesium sulphate, filtered and the volatiles evaporated to give a residue that was purified by flash chromatography on C18 silica by gradient eluition from 0 to 90%.
  • Example 27 3-(((7-(2-Aminopyrimidin-4-yl)-2,3-dihydrofuro[3,2-c]pyridin-4- yl)amino)methyl)-5-fluoro-N-(2-(2-methoxyethyl)-2-azaspiro[3.5]nonan-7- yl)benzamide (Example 27)
  • Example 27 was prepared from Intermediate 10a and 2-bromoethylmethyl ether using a procedure analogous to that used in the preparation of Example 6.
  • Example 28 3-(((7-(2-Aminopyrimidin-4-yl)-2,3-dihydrofuro[3,2-c]pyridin-4- yl)amino)methyl)-5-fluoro-N-(7-(2-methoxyethyl)-7-azaspiro[3.5]nonan-2- yl)benzamide (Example 28)
  • Example 28 was prepared from Intermediate 10b and 2-bromoethylmethyl ether using a procedure analogous to that used in the preparation of Example 6.
  • Example 29 3-(((7-(2-Aminopyrimidin-4-yl)-2,3-dihydrofuro[3,2-c]pyridin-4- yl)amino)methyl)-5-fluoro-N-(7-(oxetan-3-yl)-7-azaspiro[3.5]nonan-2-yl)benzamide (Example 29)
  • Example 29 was prepared from Intermediate 10b and 3-oxetanone using a procedure analogous to that used in the preparation of Example 19.
  • the comparative examples were prepared in a similar way to example 1 from the starting material indicated in the table below.
  • Rho kinase activity can be determined in a lOpl assay containing 40mM Tris pH7.5, 20mM MgCh O.lmg/mL BSA, 50pM DTT and 2.5pM peptide substrate (Myelin Basic Protein) using an ADP-Glo kit (Promega). Compounds were dissolved in DMSO such that the final concentration of DMSO was 1% in the assay. All reactions/incubations are performed at 25°C. Compound (2ul) and either Rho kinase 1 or 2 (4pl) were mixed and incubated for 30 mins.
  • Reactions were initiated by addition of ATP (4pl) such that the final concentration of ATP in the assay was 200pM. After a 1 hour incubation lOpl of ADP- Glo Reagent was added and after a further 1 hour incubation 20pl of Kinase Detection Buffer was added and the mixture incubated for a further 45 minutes. The luminescent signal was measured on a luminometer. Controls consisted of assay wells that did not contain compound with background determined using assay wells with no enzyme added. Compounds were tested in dose-response format and the inhibition of kinase activity was calculated at each concentration of compound.
  • the effectiveness of compounds of the present invention to inhibit PKA activity can be determined in a lOpl assay containing 40mM Tris pH7.5, 20mM MgC12 O.lmg/ml BSA, 50pM DTT and 260pM peptide substrate (kemptide) using an ADP-Glo kit (Promega).
  • Compounds were dissolved in DMSO such that the final concentration of DMSO was 1% in the assay. All reactions/incubations are performed at 25oC.
  • Compound and PKA enzyme (6pl) were mixed and incubated for 30 mins. Reactions were initiated by addition of ATP (4pl) such that the final concentration of ATP in the assay was lOpM.
  • the Compounds according to the invention showed advantageously Ki values equal to or lower than 3 nM , preferably even equal to or lower that 0,3 nM, at least on ROCK2; further preferably lower than 3 nM, preferably even equal to or lower that 0,3 nM on both isoforms.
  • the compounds according to the invention are more potent than the respective comparative example A, B and C.
  • preferred compounds according to the invention exhibit marked selectivity versus PKA.
  • the compounds according to the invention are at least 30 fold, preferably equal to or more than 100 fold, more selective in terms of ROCK2 selectivity vs PKA, than the respective comparative example A,B,C
  • mice Male CD Sprague Dawley rats (220-250 g) were purchased from Charles River Laboratories Italy (Calco, Lecco). Prior to use animals were acclimated for at least 5 days to the local vivarium conditions (room temperature: 20-24°C; relative humidity: 40-70%), having free access to standard rat chow and softened tap water. All the procedures were performed in animal operating rooms according to ethical guidelines for the conduct of animal research (D. L.vo 116/92).
  • Rats were anaesthetized with a combination of anesthetics (Zoletil 20 mg/kg + Xylazine 5 mg/kg, ip) for the i.t. administration.
  • Anesthetics Zoletil 20 mg/kg + Xylazine 5 mg/kg, ip
  • a laryngoscope was moved forward into the mouth to visualize the trachea and guide the insertion of the tip of a custom-made small diameter cannula directly into the trachea and located 1-2 mm above the bifurcation.
  • rats were surgically prepared. Body temperature was kept constant at 37° C by a heated blanket.
  • the trachea was cannulated and the lungs were ventilated artificially with a constant volume ventilator (rodent ventilator mod. 7025, Ugo Basile, Comerio, Varese, Italy) at a frequency of 80 strokes/min and at a tidal volume of 10 ml/kg.
  • a constant volume ventilator rodent ventilator mod. 7025, Ugo Basile, Comerio, Varese, Italy
  • pancuronium bromide 2 mg/kg.
  • Bronchoconstriction was induced by the i.v. injection of carbachol (cch) 80 pg/kg. In control experiments, repeated injections of this dose produced reproducible shortlasting (1-2 min duration) bronchospasms. Bronchoconstriction, quantified as a reduction of tidal volume, was evaluated according to the method described by Konzett & Roessler (1). Systemic blood pressure and changes in airway resistance were monitored with a digital pressure transducer.
  • test compounds were expressed as % inhibition of cch-evoked bronchoconstriction in time-matched, vehicle- treated, animals (controls).
  • Tested compounds were dissolved in dEEO and 1% Tween-80 or 0,001% HC1 and further diluted to target concentrations. Tested compounds were instilled locally into the trachea in a volume of 125 pl.

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Abstract

L'invention concerne des composés de formule (I) inhibant la Rho kinase qui sont des dérivés de dihydrofuropyridine, des procédés de préparation de tels composés, des compositions pharmaceutiques les contenant et leur utilisation thérapeutique. En particulier, les composés de l'invention peuvent être utiles dans le traitement de nombreux troubles associés à des mécanismes d'enzymes ROCK, y compris des maladies pulmonaires telles que l'asthme, la maladie pulmonaire obstructive chronique (MPOC), la fibrose pulmonaire idiopathique (FPI) et l'hypertension artérielle pulmonaire (HTAP).
PCT/EP2022/085286 2021-12-13 2022-12-12 Dérivés de dihydrofuropyridine utilisés comme inhibiteurs de la rho-kinase WO2023110700A1 (fr)

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