Imidazo[4,5-d]pyridazinonyl derivatives as TRPA1 inhibitors

Abstract

The present disclosure provides certain derivatives that are inhibitors of transient receptor potential ankyrin 1 (TRPA1) and are therefore useful in the treatment of diseases treatable by inhibition of TRPA1. Pharmaceutical compositions containing same and methods for preparing said compounds are also provided. (Formula (1)) wherein A is selected from the group consisting of phenyl, thiophenyl, benzothiophenyl and benzofuranyl, or A is (Formula (A)). E is selected from the group consisting of Formula (x) and Formula (y). [Formula 1] TIFF2024511181000095.tif32170 [2] TIFF2024511181000096.tif26150 [3] TIFF2024511181000097.tif39150 [C4] TIFF2024511181000098.tif39150

Description

[Technical field]

[0001] The present disclosure provides certain imidazo[4,5-d]pyridazinonyl derivatives that are inhibitors of transient receptor potential ankyrin 1 (TRPA1) and are therefore useful in treating diseases treatable by inhibition of TRPA1. Pharmaceutical compositions containing same and methods for preparing said compounds are also provided. [Background technology]

[0002] Transient receptor potential channels (TRP channels) are a group of voltage-gated ion channels located primarily in the plasma membrane of many mammalian cell types. There are approximately 30 structurally related TRP channels classified into the following groups: TRPA, TRPC, TRPM, TRPML, TRPN, TRPP, and TRPV. Transient receptor potential cation channel, subfamily A, member 1 (TRPA1), also known as transient receptor potential ankyrin 1, is the only member of the TRPA gene subfamily. Structurally, TRPA channels are characterized by multiple N-terminal ankyrin repeats (approximately 14 in the N-terminus of human TRPA1) that give rise to the "A" that stands for the ankyrin designation (Montell, 2005). TRPA1 is highly expressed in the plasma membrane of sensory neurons in dorsal root and nodose ganglia that serve both the skin and lungs, as well as in the small intestine, colon, pancreas, skeletal muscle, heart, brain, bladder and lymphocytes (https: / / www.proteinatlas.org / ), and in human lung fibroblasts.

[0003] TRPA1 is best known as a sensor for environmental stimuli that produce somatosensory modalities such as pain, cold, and itch. TRPA1 is activated by a number of reactive electrophilic stimuli (e.g., allyl isothiocyanate, reactive oxygen species) and non-reactive compounds (e.g., icilin) ​​and is involved in cough associated with asthma, chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis (IPF) or after viral infection, or in chronic idiopathic cough and cough in susceptible patients (Song and Chang, 2015; Grace and Belvisi, 2011). TRPA1 inhibitors are useful in the treatment of IPF, where cough is highly prevalent due to the association between cough and lung injury, based on studies showing cough-induced elevation of TGF-β (Xie et al., 2009; Froese et al., 2016; Tschumperlin et al., 2003; Yamamoto et al., 2002; Ahamed et al., 2008). Acute lung injury as a result of SARS-Cov-2 infection is mediated, at least in part, by reactive oxygen species (ROS). ROS are direct activators of TRPA1. Furthermore, desensitization of TRPA1 by consumption of spicy foods has been hypothesized to modulate the Nrf2 pathway and reduce oxidative stress (Bousquet et al., 2020, Bousquet et al., 2021). Thus, TRPA1 inhibitors have potential in the treatment of Covid-19 / SARS-Cov-2-induced lung injury. TRPA1 antagonists inhibit calcium signaling triggered by cough triggers such as cigarette smoke extract (CSE) oxidative stress, inflammatory mediator release, and downregulated antioxidant gene expression (Lin et al., 2015; Wang et al., 2019). TRPA1 antagonists are effective in studying atopic dermatitis (Oh et al., 2013; Wilson et al., 2013), contact dermatitis (Liu et al., 2013), psoriasis-associated pruritus (Wilson et al., 2013), and IL-31-dependent pruritus (Cevikbas et al., 2014).Human TRPA1 gain of function has been implicated in familial episodic pain syndromes (Kremeyer et al., 2010). TRPA1 antagonists were effective in a behavioral model of migraine-associated allodynia (Edelmayer et al., 2012). TRPA1 is selectively increased in the trigeminal ganglia innervating injured teeth when compared with TRPA1 expression in the trigeminal ganglia innervating healthy teeth (Haas et al., 2011). Several anesthetics are known to be TRPA1 agonists, including isoflurane (Matta et al., 2008), providing a rationale for TRPA1 inhibitors for the reduction of postoperative pain. TRPA1 knockout mice and wild-type mice treated with TRPA1 antagonists exhibited anxiolytic and antidepressant-like phenotypes (de Moura et al., 2014). TRPA1 inhibitors are expected to have benefit in the treatment of diabetic neuropathy based on studies showing a mechanistic link between AMPK and TRPA1 inverse regulation (Hiyama et al., 2018; Koivisto and Pertovaara, 2013; Wang et al., 2018). TRPA1 knockout mice exhibit smaller myocardial infarct size compared to wild-type mice (Conklin et al., 2019). TRPA1 knockout and pharmacological intervention inhibited TNBS-induced colitis in mice (Engel et al., 2011). In a mouse cerebral ischemia model, TRPA1 knockout and TRPA1 antagonists reduce myelin damage (Hamilton et al., 2016). Uric acid crystals and joint inflammation are reduced in TRPA1 knockout mice in the monosodium urate mouse model of gout (Moilanen et al., 2015). TRPA1 deletion in rats ameliorates joint inflammation and hyperalgesia in a rat model of acute gout flare ( Trevisan et al., 2014 ). Activation of TRPA1 induces an inflammatory response in osteoarthritis chondrocytes ( Nummenmaa et al., 2016 ).TRPA1 inhibition and genetic deletion reduce inflammatory mediators in osteoarthritic mouse chondrocytes and murine cartilage (Nummenmaa et al., 2016). Finally, TRPA1 knockout mice exhibit improved weight bearing on the osteoarthritic limb in an MIA-induced knee swelling model (Horvath et al., 2016). TRPA1 is differentially expressed in the bladder epithelium of rats (Du et al., 2007) and in patients with bladder outlet obstruction (Du et al., 2008). TRPA1 receptor modulation attenuates bladder overactivity in a rat model of spinal cord injury (Andrade et al., 2011), and intrathecal administration of a TRPA1 antagonist attenuates cyclophosphamide-induced cystitis in rats with micturition hyperreflexia (Chen et al., 2016). Summary of the Invention

[0004] It would therefore be desirable to provide potent TRPA1 inhibitors. TRPA1 inhibitors of different structural classes are reviewed in S. Skerratt, Progress in Medicinal Chemistry, 2017, Volume 56, 81-115, D. Preti, G. Saponaro, A. Szallasi, Pharm. Pat. Anal. (2015) 4 (2), 75-94, and H. Chen, Transient receptor potential ankyrin 1 (TRPA1) antagonists: a patent review (2015-2019), Expert Opin Ther Pat., 2020. WO 2017 / 060488 discloses a compound having the generalized structural formula [ka] The present invention discloses compounds that are antagonists of TRPA1, having the formula:

[0005] Among them, IC<100 nM in calcium influx assay 50 The TRPA1 activity of Examples 53, 72, 73, 86 and 90 is disclosed. L. Schenkel, et al., J. Med. Chem. 2016, 59, 2794-2809, has the generalized structure [ka] The present invention discloses quinazolinone-based TRPA1 antagonists, including compounds of the formula: 50 and is disclosed as having an intrinsic clearance of less than 14 μL / min / kg in human liver microsomes. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0006] The present invention discloses novel derivatives that are inhibitors of Transient Receptor Potential Ankyrin 1 (TRPA1) and that possess suitable pharmacological and pharmacokinetic properties that allow their use as medicines for the treatment of conditions and / or diseases treatable by inhibition of TRPA1. The compounds of the present invention may offer several advantages, such as enhanced potency, increased metabolic and / or chemical stability, increased selectivity, safety and tolerability, enhanced solubility, enhanced permeability, favorable plasma protein binding, enhanced bioavailability, favorable pharmacokinetic profiles, and the potential to form stable salts. Compounds of the Invention The present invention is a surprisingly potent inhibitor of TRPA1 (Assay A), - Improved stability in human liver microsomes (Assay B) - Improved stability in human hepatocytes (Assay C) The present invention provides a derivative further characterized by:

[0007] The compounds of the present invention are structurally distinct from Examples 53, 72, 73, 86 and 90 of WO 2017 / 060488 and Example 31 of L. Schenkel, et al., J. Med. Chem. 2016, 59, 2794-2809 in that they contain a substituted imidazo[4,5-d]pyridazinonyl core and substituents adjacent to the secondary aliphatic alcohol. These structural differences unexpectedly lead to a favorable combination of (i) inhibition of TRPA1, (ii) stability in human liver microsomes, and (iii) stability in human hepatocytes. Stability in human liver microsomes refers to the susceptibility of a compound to biotransformation in the context of selecting and / or designing drugs with favorable pharmacokinetic properties as a first screening step. The main metabolic site for many drugs is the liver. Human liver microsomes contain cytochrome P450 (CYP) and therefore represent a model system for studying phase I drug metabolism in vitro. Enhanced stability in human liver microsomes is associated with several advantages, including increased bioavailability and sufficient half-life, which can allow less frequent dosing of patients. Therefore, enhanced stability in human liver microsomes is a favorable feature for compounds used in drugs. Thus, it is expected that the compounds of the present invention, in addition to being able to inhibit TRPA1, will have favorable in vivo clearance and therefore a desired duration of action in humans.

[0008] Stability in human hepatocytes refers to the susceptibility of a compound to biotransformation in the context of selecting and / or designing drugs with favorable pharmacokinetic properties. The primary metabolic site for many drugs is the liver. Human hepatocytes contain cytochrome P450 (CYP) and other drug metabolizing enzymes and therefore represent a model system for studying drug metabolism in vitro (importantly, in contrast to liver microsomal assays, hepatocyte assays also encompass phase II biotransformation and liver-specific transporter-mediated processes and therefore represent a more complete system for drug metabolism studies). Enhanced stability in human hepatocytes is associated with several advantages, including increased bioavailability and sufficient half-life, which can allow for less frequent dosing of patients. Therefore, enhanced stability in human hepatocytes is a favorable feature for compounds used in drugs.

[0009] The present invention relates to novel compounds according to formula (I) [ka] (In the formula, A is selected from the group consisting of phenyl, thiophenyl, benzothiophenyl, and benzofuranyl, where A is unsubstituted or selected from halogen and C 1-4 -alkyl radical R 1 or is replaced by one or two members of Or, A is

[0010] [ka] and E is [ka] (selected from the group consisting of to provide.

[0011] Another embodiment of the invention is a phenyl group selected from the group consisting of phenyl, thiophenyl, benzothiophenyl and benzofuranyl, where A is unsubstituted or is a group R consisting of Br, Cl, F and HC. 1 or is replaced by one or two members of Or, A: [ka] and The substituent E is defined as in the preceding embodiment. Concerning compounds of formula (I):

[0012] Another embodiment of the present invention is where A is selected from the group consisting of phenyl, benzothiophenyl and benzofuranyl, where A is unsubstituted or selected from halogen and C 1-4 -alkyl radical R 1 or is replaced by one or two members of Or, A: [ka] and The substituent E is defined as in any of the preceding embodiments. Concerning compounds of formula (I):

[0013] Another embodiment of the invention is a phenyl group, wherein A is selected from the group consisting of phenyl, benzothiophenyl and benzofuranyl, where A is unsubstituted or is a group R consisting of Br, Cl, F and HC. 1 or is replaced by one or two members of Or, A: [ka] and The substituent E is defined as in any of the preceding embodiments. Concerning compounds of formula (I):

[0014] Another embodiment of the present invention is wherein A is [ka] where A is unsubstituted or a group R 1 or is replaced by one or two members of Or, A, [ka] and Substituents E and R 1 is defined as in any of the preceding embodiments. Concerning compounds of formula (I):

[0015] Another embodiment of the present invention comprises: A, [ka] [ka] is selected from the group consisting of The substituent E is defined as in any of the preceding embodiments. Concerning compounds of formula (I):

[0016] Another embodiment of the present invention is wherein E is [ka] and substituent A is defined as in any of the preceding embodiments. Concerning compounds of formula (I):

[0017] Another embodiment of the present invention is wherein E is [ka] and substituent A is defined as in any of the preceding embodiments. Concerning compounds of formula (I): Particularly preferred are

[0018] [ka] [ka] [ka] The compound according to formula (I) is selected from the group consisting of:

[0019] Terms and definitions used Terms not specifically defined herein should be given the meaning that would be given to them by one of ordinary skill in the art in light of the present disclosure and the context. However, as used herein, unless expressly stated to the contrary, the following terms have the meaning indicated and are subject to the following contractions: In the groups, radicals or moieties defined below, the number of carbon atoms is often specified preceding the group, e.g., C 1-6 -Alkyl means an alkyl group or radical having 1 to 6 carbon atoms. Generally, in groups such as HO, H2N, (O)S, (O)2S, NC (cyano), HOOC, F3C, etc., one skilled in the art can see the point of radical attachment to the molecule from the free valence of the group itself. For composite groups containing two or more subgroups, the last listed subgroup is the point of radical attachment, e.g., the substituent "aryl-C 1-3 -alkyl" is C 1-3 - means an aryl group bonded to an alkyl group, the latter being bonded to the core or to a group to which a substituent is bonded.

[0020] When the compounds of the invention are depicted in the form of a chemical name and as a formula, in the event of any discrepancy, the formula shall prevail. An asterisk may be used in a subformula to indicate a bond that is connected to the core molecule as defined. Numbering of substituent atoms begins with the atom closest to the core or to the group to which the substituent is attached. For example, the term "3-carboxypropyl group" refers to the following substituent: [ka] where the carboxy group is attached to the third carbon atom of the propyl group. The term "1-methylpropyl-", "2,2-dimethylpropyl-" or "cyclopropylmethyl-" refers to the following group:

[0021] [ka] An asterisk may be used in a sub-formula to indicate a bond that is connected to the core molecule as defined.

[0022] The term “C 1-n -alkyl" (wherein n is an integer selected from 2, 3, 4 or 5) denotes, either alone or in combination with another radical, an acyclic, saturated, branched or linear hydrocarbon radical having 1 to n C atoms. For example, C 1-5 The term -alkyl includes the radicals HC-, HC-CH-, HC-CH-CH-, HC-CH(CH)-, HC-CH-CH-CH-, HC-CH(CH)-, HC-CH-CH-CH-, HC-CH(CH)-, HC-CH(CH)-CH-, HC-C(CH)-, HC-CH-CH-CH-CH-, HC-CH-CH-CH-, HC-CH-CH-CH-, HC-CH-CH-CH-, HC-CH-CH-CH-, HC-CH-CH-CH-, HC-CH-CH-CH-, HC-CH-C(CH)-, HC-C(CH)-, HC-CH(CH)-CH-, HC-CH-CH-CH- and HC-CH-CH(CHCH)-. The term "fluoro" (saturated or unsaturated) appended to an "alkyl," "alkylene," or "cycloalkyl" group means such an alkyl or cycloalkyl group in which one or more hydrogen atoms have been replaced by a fluorine atom. Examples include, but are not limited to, H2FC-, HF2C-, and F3C-. The term phenyl refers to the following ring:

[0023] [ka] Refers to the radical of The term thiophenyl refers to the following ring: [ka] Refers to the radical of

[0024] The term benzofuranyl refers to the following ring: [ka] Refers to the radical of The term benzothiophenyl refers to the following ring: [ka] Refers to the radical of

[0025] The term imidazo[4,5-d]pyridazinonyl refers to the following bicyclic core: [ka] Refers to the radical of The term "substituted," as used herein, means that any one or more hydrogens on the designated atom are replaced with a selection from the indicated group, provided that the replacement does not exceed the normal valence of the designated atom, and that the substitution results in a stable compound.

[0026] Unless specifically indicated, throughout this specification and the appended claims, a given chemical formula or name is intended to encompass tautomers and all stereo, optical and geometric isomers (e.g., enantiomers, diastereomers, E / Z isomers, etc.) and their racemates, as well as mixtures of different proportions of the separate enantiomers, mixtures of diastereomers, or mixtures of any of the foregoing forms when such isomers and enantiomers exist, as well as pharmaceutically acceptable salts thereof, and solvates thereof, such as hydrates, including solvates of the free compounds or solvates of the salts of the compounds. In general, substantially pure stereoisomers may be obtained according to synthetic principles known to those skilled in the art, for example, by separation of corresponding mixtures, by using stereochemically pure starting materials, and / or by stereoselective synthesis. It is known in the art how to prepare optically active forms, such as by resolution of racemic forms or by synthesis starting from, for example, optically active starting materials, and / or by using chiral reagents. Enantiomerically pure compounds or intermediates of the present invention may be prepared via asymmetric synthesis, for example by the preparation and subsequent separation of suitable diastereomeric compounds or intermediates which may be separated by known methods (e.g. by chromatographic separation or crystallization), and / or by using chiral reagents, such as chiral starting materials, chiral catalysts, or chiral auxiliaries.

[0027] Furthermore, it is known to those skilled in the art how to prepare enantiomerically pure compounds from the corresponding racemic mixtures, such as by chromatographic separation of the corresponding racemic mixtures on chiral stationary phases; or by resolution of the racemic mixtures using a suitable resolving agent, for example by means of diastereomeric salt formation of the racemates with an optically active acid or base, followed by resolution of the salts and release of the desired compound from the salt; or by derivatization of the corresponding racemate with an optically active chiral auxiliary reagent, followed by diastereomeric separation and removal of the chiral auxiliary; or by kinetic resolution of the racemates (e.g. by enzymatic resolution); by enantioselective crystallization from a consortium of enantiomeric crystals under suitable conditions; or by (fractional) crystallization from a suitable solvent in the presence of an optically active chiral auxiliary, etc. The phrase "pharmacologically acceptable" is used herein to refer to those compounds, materials, compositions and / or dosage forms that are suitable for use without excessive toxicity, irritation, allergic response or other problem or complication, within the scope of sound medical judgment, commensurate with a reasonable benefit / risk ratio.

[0028] As used herein, "pharmaceutically acceptable salts" refer to derivatives of the disclosed compounds where the parent compound forms salts or complexes with acids or bases. Examples of acids that form pharma- ceutically acceptable salts with the parent compound that contains a basic moiety include mineral or organic acids, such as 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. Examples of cations and bases that form pharma- ceutically acceptable salts with the parent compound that contains an acidic moiety are Na + , K + , Ca 2+ , Mg 2+ , NH4 + , L-arginine, 2,2'-iminobisethanol, L-lysine, N-methyl-D-glucamine or tris(hydroxymethyl)-aminomethane. The pharma- ceutically acceptable salts of the present invention can be synthesized from the parent compound that contains a basic or acidic moiety by conventional chemical methods. In general, such salts can be prepared by reacting the free acid or base form of these compounds with a sufficient amount of an appropriate base or acid in water or an organic diluent such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile or a mixture thereof. Salts of acids other than those mentioned above, which are useful, for example, for purifying or isolating the compounds of the invention (eg trifluoroacetates), also form part of the invention.

[0029] Biological assays Assessment of TRPA1 activity Assay A: TRPA1 assay The activity of the compounds of the invention can be demonstrated using the following in vitro TRPA1 cellular assay. method: Human HEK293 cell line (Perkin Elmer, product number AX-004-PCL) overexpressing human TRPA1 ion channel is used as a test system for compound efficacy and potency. Compound activity is determined by measuring the effect of compound on intracellular calcium concentration induced by AITC (Allylisothiocyanat) agonism in FLIPR Tetra system (Molecular Devices). Cell culture: Cells are obtained as frozen cells in cryovials and stored at -150°C until use. Cells are grown in culture medium (MEM / EBSS medium with 10% FCS and 0.4mg / ML Geneticin). It is important that the density does not exceed 90% confluence. For subculture, cells are detached from flasks with Versen's solution. The day before the assay, cells are detached, washed twice with medium (MEM / EBSS medium with 10% FCS) and seeded at 20000 cells in 20μl / well in Corning poly-D-lysine biocoated 384-well plates (black, clear bottom, catalogue no. 356697). Plates are incubated at 37°C / 5% CO2 for 24 hours before use in the assay.

[0030] compound preparation Test compounds are dissolved in 100% DMSO at a concentration of 10 mM, diluted in DMSO at a concentration of 5 mM in a first step, followed by serial dilution steps in 100% DMSO. Dilution ratios and number of dilution steps can vary as required. Typically 8 different concentrations are prepared at 1:5 dilution, and further intermediate dilutions (1:20) of substances are made using HBSS / HEPES buffer (1xHEPES from Gibco, Cat. No. 14065; 20mM HEPES from SIGMA, Cat. No. 83264; 0.1%BSA from Invitrogen, Cat. No. 11926, pH 7.4).

[0031] FLIPR Assay: On the day of the assay, the cells are washed three times with assay buffer (puffer), leaving 20 μL of buffer in the well after washing. 10 μL of Ca6 kit (catalog no. R8191 MolecularDevices) loading buffer in HBSS / HEPES is added to the cells, the plate is covered and incubated at 37° C. / 5% CO2 for 120 minutes. 10 μL of compound or control in HBSS / HEPES buffer / 5% DMSO is carefully added to the wells from the intermediate dilution plate. Luminescence (indicative of calcium influx or release) is read on the FLIPR Tetra device for 10 minutes to monitor compound-induced effects (e.g. agonism). Finally, 10 μL of the agonist AITC 50 μM dissolved in HBSS / HEPES buffer / 0.05% DMSO (final concentration 10 μM) is added to the wells, followed by an additional reading on the FLIPR Tetra device for 10 minutes. The area under the signal curve (AUC) after addition of AITC is used to calculate IC50 / % inhibition.

[0032] Data evaluation and calculations: Each assay microtiter plate contains wells with vehicle (1% DMSO) control instead of compound (100% CTL; high control) as a control for AITC-induced luminescence, and wells with vehicle control without AITC (0% CTL; low control) as a control for non-specific changes in luminescence. The analysis of the data is carried out by calculation of the area under the signal curve for the individual wells. Based on this value, the MegaLab software (in-house development) is used to calculate the % value for each substance concentration measurement (AUC(sample)-AUC(low)) x 100 / (AUC(high)-AUC(low)). The IC50 value is calculated from the control % value using the MegaLab software. Calculation: [y=(ad) / (1+(x / c)^b)+d], a=low value, d=high value; x=concentration M; c=IC50M; b=Hill; y=control %

[0033] [Table 1]

[0034] [Table 2]

[0035] [Table 3]

[0036] Assessment of microsomal clearance Assay B: Microsomal Clearance: The metabolic degradation of test compounds is assayed using pooled liver microsomes at 37° C. A final incubation volume of 100 μl per time point contains TRIS buffer pH 7.6 (0.1 M), magnesium chloride (5 mM), microsomal protein (1 mg / ml) and test compound at a final concentration of 1 μM at room temperature. After a short preincubation period at 37°C, the reaction is started by the addition of beta-nicotinamide adenine dinucleotide phosphate, reduced form (NADPH, 1 mM) and terminated by transferring aliquots into the solvent after different time points (0, 5, 15, 30, 60 min). In addition, NADPH-independent degradation is monitored by incubation without NADPH and terminated at the final time point. The remaining test compound [%] after NADPH-independent incubation is reflected by the parameter c(control) (metabolic stability). The quenched incubation is pelleted by centrifugation (10000g, 5 min). An aliquot of the supernatant is assayed for the amount of parent compound by LC-MS / MS. The half-life (t1 / 2 INVITRO) is determined by the slope of a semi-log plot of the concentration-time profile.

[0037] The intrinsic clearance (CL_INTRINSIC) is calculated by taking into account the amount of protein in the incubation: CL_INTRINSIC [μl / min / mg protein] = (Ln2 / (half-life [min] × protein content [mg / ml])) × 1000 CL_INTRINSIC_INVIVO[ml / min / kg]=(CL_INTRINSIC[μL / min / mg protein]×MPPGL[mg protein / g liver]×hepatic factor[g / kg body weight]) / 1000 Qh[%]=CL[ml / min / kg] / hepatic blood flow [ml / min / kg]) Hepatocellularity, human: 120 x 10e6 cells / g liver Hepatic factor, human: 25.7 g / kg body weight Blood flow rate, human: 21 ml / (min x kg)

[0038] [Table 4]

[0039] [Table 5]

[0040] [Table 6]

[0041] Assessment of hepatocyte clearance Assay C: Hepatocyte Clearance The metabolic degradation of the test compounds is assayed in hepatocyte suspensions: Hepatocytes (cryopreserved) are incubated in Dulbecco's modified Eagle's medium (supplemented with 3.5 μg glucagon / 500 mL, 2.5 mg insulin / 500 mL and 3.75 mg / 500 mL hydrocortisone) containing 5% seed serum. After 30 min preincubation in an incubator (37 °C, 10% CO), add 5 µl of test compound solution (80 µM; from 2 mM in DMSO stock solution diluted 1:25 with culture medium) into 395 µl of hepatocyte suspension (cell density in the range of 0.25-5 million cells / mL depending on species, typically 1 million cells / mL; final concentration of test compound 1 µM, final DMSO concentration 0.05%). Cells are incubated (incubator, orbital shaker) for 6 hours and samples (25 μl) are taken at 0, 0.5, 1, 2, 4 and 6 hours. Samples are transferred into acetonitrile and pelleted by centrifugation (5 min). Supernatants are transferred to a new 96 deep-well plate, evaporated under nitrogen and resuspended. The loss of parent compound is analyzed by HPLC-MS / MS. CLint is calculated as follows: CL_INTRINSIC=dose / AUC=(C0 / CD) / (AUD+clast / k)×1000 / 60. C0: initial concentration in incubation [μM], CD: cell density of viable cells [10e6 cells / mL], AUD: area under the data [μM×hour], clast: concentration of final data point [μM], k: slope of regression line for parental decline [hour-1].

[0042] The calculated in vitro hepatic intrinsic clearance can be scaled up to intrinsic in vivo hepatic clearance and used to predict hepatic in vivo blood clearance (CL) by use of a liver model (well-stirred model). CL_INTRINSIC_INVIVO[ml / min / kg]=(CL_INTRINSIC[μL / min / 10e6 cells]×hepatocyte fullness [10e6 cells / g liver]×liver factor [g / kg body weight]) / 1000 CL[ml / min / kg]=CL_INTRINSIC_INVIVO[ml / min / kg]×hepatic blood flow [ml / min / kg] / (CL_INTRINSIC_INVIVO[ml / min / kg]+hepatic blood flow [ml / min / kg]) Qh[%]=CL[ml / min / kg] / hepatic blood flow [ml / min / kg]) Liver cellularity, human: 120 x 10e6 cells / g liver Hepatic factor, human: 25.7 g / kg body weight Blood flow rate, human: 21 ml / (min x kg)

[0043] [Table 7]

[0044] [Table 8]

[0045] [Table 9]

[0046] Permeability assessment Caco-2 cells (1-2 x 105 cells / cm2 area) are seeded onto filter inserts (Costar Transwell polycarbonate or PET filters, 0.4 μm pore size) and cultured (DMEM) for 10-25 days. Compounds are dissolved in an appropriate solvent (such as DMSO, 1-20 mM stock solutions). Stock solutions are diluted with HTP-4 buffer (128.13 mM NaCl, 5.36 mM KCl, 1 mM MgSO4, 1.8 mM CaCl2, 4.17 mM NaHCO3, 1.19 mM Na2HPO4 x 7H2O, 0.41 mM NaH2PO4 x 7H2O, 15 mM HEPES, 20 mM glucose, 0.25% BSA, pH 7.2) to prepare transport solution (0.1-300 μM compound, final DMSO ≤ 0.5%). Transport solution (TL) is applied to the apical or basolateral donor side to measure AB or BA permeability (filter triplicate), respectively. Samples are collected from the donor at the beginning and end of the experiment and also from the receiver side at various time intervals up to 2 hours for concentration measurement by HPLC-MS / MS or scintillation counting. The sampled receiver volume is replaced with fresh receiver solution.

[0047] Assessment of plasma protein binding This equilibrium dialysis (ED) technique is used to determine the approximate in vitro fractional binding of test compounds with plasma proteins. Dianorm Teflon dialysis cells (micro 0.2) are used. Each cell consists of a donor and an acceptor chamber separated by an ultrathin semipermeable membrane with a molecular weight cut-off of 5 kDa. Stock solutions of each test compound are prepared at 1 mM in DMSO and diluted to a final concentration of 1.0 μM. Subsequent dialysis solutions are prepared in pooled human or rat plasma (with NaEDTA added) from male and female donors. An aliquot of 200 μL of dialysis buffer (100 mM potassium phosphate, pH 7.4) is dispensed into the buffer chamber. An aliquot of 200 μL of test compound dialysis solution is dispensed into the plasma chamber. Incubation is performed for 2 hours at 37° C. with rotation. At the end of the dialysis period, the dialysate is transferred to a reaction tube. The tube for the buffer fraction contains 0.2 mL of ACN / water (80 / 20). An aliquot of 25 μL of plasma dialysate is transferred to a deep well plate and mixed with 25 μL of ACN / water (80 / 20), 25 μL of buffer, 25 μL of calibration solution and 25 μL of internal standard solution. Protein precipitation is performed by adding 200 μL of ACN. An aliquot of 50 μL of buffer dialysate is transferred to a deep well plate and mixed with 25 μL of blank plasma, 25 μL of internal standard solution and 200 μL of ACN. The samples are measured in an HPLC-MS / MS system and evaluated in the analyst software. The binding percentage is calculated by the formula: % bound = (plasma concentration - buffer concentration / plasma 30 concentration) x 100.

[0048] Solubility evaluation Saturated solutions are prepared in a well plate (format determined by the robot) by adding an appropriate volume of the selected aqueous medium (typically in the range of 0.25-1.5 ml) to each well containing a known amount of solid drug substance (typically in the range of 0.5-5.0 mg). The wells are shaken or stirred for a predefined period (typically in the range of 2-24 h) and then filtered using a suitable filter membrane (typically a PTFE filter with a pore size of 0.45 μm). Filter absorption is avoided by discarding the first few drops of the filtrate. The amount of dissolved drug substance is determined by UV spectroscopy. In addition, the pH of the saturated aqueous solution is measured using a glass electrode pH meter.

[0049] Pharmacokinetic characterization Test compounds are administered to each test species either intravenously or orally. Blood samples are taken at several time points after application of the test compound, anticoagulated, and centrifuged. Test substance concentrations - administered compounds and / or metabolites - are quantified in plasma samples. PK parameters are calculated using non-compartmental methods. AUC and Cmax are normalized to a dose of 1 μmol / kg. Assessment of metabolism in human hepatocytes in vitro The metabolic pathway of the test compounds is investigated using primary human hepatocytes in suspension. After recovery from cryopreservation, the human hepatocytes are incubated in Dulbecco's modified Eagle's medium containing 5% human serum and supplemented with 3.5 μg glucagon / 500 ml, 2.5 mg insulin / 500 ml, and 3.75 mg / 500 ml hydrocortisone. After preincubation for 30 min in a cell culture incubator (37°C, 10% CO2), the test compound solution was mixed into the hepatocyte suspension and diluted to 1.0 × 10 6 ~4.0×10 6 A final cell density of cells / ml (according to compound turnover rates observed in primary human hepatocytes), a final test compound concentration of 10 μM and a final DMSO concentration of 0.05% are obtained.

[0050] Cells are incubated for 6 h in a cell culture incubator on a horizontal shaker and samples are removed from incubation after 0, 0.5, 1, 2, 4 or 6 h depending on turnover rate. Samples are quenched with acetonitrile and pelleted by centrifugation. Supernatants are transferred to 96 deep-well plates, evaporated under nitrogen, resuspended and then bioanalyzed by liquid chromatography-high resolution mass spectrometry for identification of putative metabolites. Fourier Transform MS n Structures are tentatively assigned based on the data. Metabolites are reported as percentages of the parent in human hepatocyte incubations with a threshold of 4% or higher.

[0051] Treatment The present invention relates to compounds of general formula 1 useful in the prevention and / or treatment of diseases and / or conditions associated with or modulated by TRPA1 activity, including, but not limited to, fibrotic diseases, inflammatory and immunoregulatory disorders, respiratory or gastrointestinal diseases or complaints, ophthalmological diseases, inflammatory diseases of the joints and inflammatory diseases of the nasopharynx, eyes and skin, and pain and neurological disorders, including cough, idiopathic pulmonary fibrosis, other interstitial lung diseases and other fibrotic, asthmatic or allergic diseases, eosinophilic diseases, chronic obstructive pulmonary disease, and inflammatory and immunoregulatory disorders, such as rheumatoid arthritis and atherosclerosis, and pain and neurological disorders, such as acute pain, surgical pain, chronic pain, and depression and bladder disorders.

[0052] The compounds of general formula 1 are useful for the prevention and / or treatment of: (1) Cough, including, for example, chronic idiopathic cough or chronic refractory cough, cough associated with asthma, COPD, lung cancer, post-viral and idiopathic pulmonary fibrosis, and other interstitial lung diseases. (2) Pulmonary fibrotic diseases, for example, pneumonia or interstitial pneumonia associated with collagen diseases, such as lupus erythematosus, systemic sclerosis, rheumatoid arthritis, polymyositis and dermatomysitis, idiopathic interstitial pneumonia, for example, pulmonary fibrosis Interstitial lung disease (IPF), nonspecific interstitial pneumonia, respiratory bronchiolitis-associated interstitial lung disease, desquamative interstitial pneumonia, cryptogenic organizing pneumonia, acute interstitial pneumonia and lymphocytic interstitial pneumonia, lymphangioleiomyomatosis, pulmonary alveolar proteinosis, Langerhans cell histiocytosis, pleuroparenchymal fibroelastosis, interstitial lung disease of known etiology, for example asbestosis, silicosis, miner's lung (coal dust), farmer's lung (hay and mold), pigeon breeder's lung (birds), or other occupational airborne triggers such as metal dust or mycobacteria. Interstitial pneumonia as a result of occupational exposure such as pulmonary embolism, pulmonary embolism, pulmonary fibrosis, acute lung injury as a result of Covid-19 / SARS-Cov-2 infection or pulmonary fibrosis secondary to Covid-19 / SARS-Cov-2 infection, hypersensitivity pneumonitis or interstitial pneumonia caused by aspiration, inhalation of toxic gases, vapors, heart failure, X-rays, radiation, chemotherapy, M. boeck or sarcoidosis, granulomatosis, cystic fibrosis or cystic fibrosis, alpha-1-antitrypsin deficiency, interstitial pneumonia as a result of occupational exposure such as pulmonary embolism, pulmonary embolism, pulmonary fibrosis secondary to Covid-19 / SARS-Cov-2 infection, or as a result of treatment for granulomatous diseases such as granulomatosis with polyangiitis, Churg-Strauss syndrome, sarcoidosis, bronchitis or pneumonia or interstitial pneumonia caused by acute lung injury as a result of Covid-19 / SARS-Cov-2 infection or pulmonary fibrosis secondary to Covid-19 / SARS-Cov-2 infection. (3) Other fibrotic diseases, such as hepatic bridging fibrosis, cirrhosis, nonalcoholic steatohepatitis (NASH), atrial fibrosis, myocardial fibrosis, old myocardial infarction, glial scar, arterial sclerosis, articular fibrosis, Dupuytren's contracture, keloids, scleroderma / systemic sclerosis, mediastinal fibrosis, myelofibrosis, Peyronie's disease, nephrogenic systemic fibrosis, retroperitoneal fibrosis, and adhesive capsulitis.

[0053] (4) Inflammatory, autoimmune or allergic diseases and conditions, such as allergic or nonallergic rhinitis or sinusitis, chronic sinusitis or rhinitis, nasal polyposis, chronic rhinosinusitis, acute rhinosinusitis, asthma, childhood asthma, allergic bronchitis, alveolitis, hyperreactive airways, allergic conjunctivitis, bronchiectasis, adult respiratory distress syndrome, bronchial and pulmonary edema, bronchitis or pneumonia, eosinophilic cellulites (e.g., Wells' syndrome), eosinophilic pneumonia (e.g., Löffler's syndrome, chronic eosinophilic pneumonia), eosinophilic fasciitis (e.g., Schulman's syndrome), delayed hypersensitivity, non-allergic asthma; exercise-induced bronchoconstriction; chronic obstructive pulmonary disease (COPD), acute bronchitis, chronic bronchitis, cough, emphysema; systemic anaphylaxis or hypersensitivity reactions, drug allergies (e.g., to penicillin, cephalosporins), eosinophilia myalgia syndrome due to ingestion of contaminated tryptophan, insect bite allergies; autoimmune diseases, such as Rheumatoid arthritis, Graves' disease, Sjogren's syndrome, psoriatic arthritis, multiple sclerosis, systemic lupus erythematosus, myasthenia gravis, immune thrombocytopenia (adult ITP, neonatal thrombocytopenia, pediatric ITP), immune hemolytic anemia (autoimmune and drug-induced), Evans syndrome (platelet and red blood cell immune cytopenia), neonatal Rh disease, Goodpasture's syndrome (anti-GBM disease), celiac disease, autoimmune cardiomyopathy, juvenile onset diabetes mellitus; glomerulonephritis, autoimmune thyroiditis, Bethesda syndrome, pulmonary arthritis; transplant rejection (e.g. in transplant surgery), including allograft rejection or graft-versus-host disease; inflammatory bowel diseases, such as Crohn's disease and ulcerative colitis; spondyloarthropathy; scleroderma; psoriasis (including T-cell mediated psoriasis) and inflammatory skin diseases, such as dermatitis, eczema, atopic dermatitis, allergic contact dermatitis, urticaria; vasculitis (e.g. necrotizing, cutaneous and hypersensitivity vasculitis); erythema nodosum; eosinophilic myositis, eosinophilic fasciitis, cancer with leukocyte infiltration of the skin or organs;Ophthalmic diseases, such as age-related macular degeneration, diabetic retinopathy and diabetic macular edema, keratitis, eosinophilic keratitis, keratoconjunctivitis, vernal conjunctivitis, scars, anterior eye scars, blepharitis, blepharoconjunctivitis, bullous disorders, cicatricial pemphigoid, conjunctival melanoma, papillary conjunctivitis, dry eye, episcleritis, glaucoma, gliosis, granuloma annulare, Graves' ophthalmopathy, intraocular melanoma, pinguecula, proliferative vitreoretinopathy, pterygium, scleritis, uveitis, acute gout flare, gout or osteoarthritis;

[0054] (5) Pain, for example, chronic idiopathic pain syndromes, neuropathic pain, paresthesia, allodynia, migraine, dental pain and post-operative pain. (6) Depression, anxiety, diabetic neuropathy and bladder disorders, e.g., bladder outlet obstruction, overactive bladder, cystitis; myocardial reperfusion injury or cerebral ischemic injury. Thus, the present invention relates to compounds of general formula 1 for use as medicaments. Furthermore, the present invention relates to the use of compounds of general formula 1 for the treatment and / or prevention of diseases and / or conditions associated with or modulated by TRPA1 activity.

[0055] Furthermore, the present invention relates to the use of compounds of general formula 1 for the treatment and / or prevention of fibrotic diseases, inflammatory and immunoregulatory disorders, respiratory or gastrointestinal diseases or complaints, ophthalmological diseases, inflammatory diseases of the joints and inflammatory diseases of the nasopharynx, eyes and skin, as well as pain and neurological disorders, including cough, idiopathic pulmonary fibrosis, other interstitial lung diseases and other fibrotic, asthmatic or allergic diseases, eosinophilic diseases, chronic obstructive pulmonary disease, and inflammatory and immunoregulatory disorders, such as rheumatoid arthritis and atherosclerosis, as well as pain and neurological disorders, such as acute pain, surgical pain, chronic pain, as well as depression and bladder disorders. Furthermore, the present invention relates to the use of compounds of general formula 1 for the treatment and / or prevention of:

[0056] (1) Cough, including, for example, chronic idiopathic cough or chronic refractory cough, cough associated with asthma, COPD, lung cancer, post-viral and idiopathic pulmonary fibrosis, and other interstitial lung diseases. (2) Pulmonary fibrotic diseases, e.g., pneumonia or interstitial pneumonia associated with collagen diseases, e.g., lupus erythematosus, systemic sclerosis, rheumatoid arthritis, polymyositis and dermatomyositis, idiopathic interstitial pneumonias, e.g., pulmonary fibrosis (IPF), nonspecific interstitial pneumonia, respiratory bronchiolitis-associated interstitial lung disease, desquamative interstitial pneumonia, idiopathic organizing pneumonia, acute interstitial pneumonia and lymphocytic interstitial pneumonia, lymphangioleiomyomatosis, pulmonary alveolar proteinosis, Langerhans cell histiocytosis, pleuroparenchymal fibroelastosis, interstitial lung diseases of known etiology, e.g., asbestosis, silicosis, miner's lung (coal dust), farmer's lung (hay and mold), pigeon breeder's lung (birds), or other occupational airborne triggers, such as metal dust or mycobacteria, as a result of occupational exposure, or radiation Interstitial pneumonia, hypersensitivity pneumonitis, or interstitial pneumonia of different origins, e.g. caused by aspiration, inhalation of toxic gases, vapors, as a result of treatments for granulomatous diseases such as methotrexate, amiodarone, nitrofurantoin or chemotherapy drugs, or for granulomatous diseases such as granulomatosis with polyangiitis, Churg-Strauss syndrome, sarcoidosis, bronchitis or pneumonia or interstitial pneumonia caused by heart failure, x-rays, radiation, chemotherapy, M. boeck or sarcoidosis, granulomatosis, cystic fibrosis or cystic fibrosis, alpha-1-antitrypsin deficiency, acute lung injury as a result of Covid-19 / SARS-Cov-2 infection or pulmonary fibrosis secondary to Covid-19 / SARS-Cov-2 infection. (3) Other fibrotic diseases, such as bridging fibrosis in the liver, cirrhosis, nonalcoholic steatohepatitis (NASH), atrial fibrosis, myocardial fibrosis, old myocardial infarction, glial scar, arterial sclerosis, articular fibrosis, Dupuytren's contracture, keloids, scleroderma / systemic sclerosis, mediastinal fibrosis, myelofibrosis, Peyronie's disease, nephrogenic systemic fibrosis, retroperitoneal fibrosis, and adhesive capsulitis.

[0057] (4) Inflammatory, autoimmune or allergic diseases and conditions, such as allergic or nonallergic rhinitis or sinusitis, chronic sinusitis or rhinitis, nasal polyposis, chronic rhinosinusitis, acute rhinosinusitis, asthma, childhood asthma, allergic bronchitis, alveolitis, hyperreactive airways, allergic conjunctivitis, bronchiectasis, adult respiratory distress syndrome, bronchial and pulmonary edema, bronchitis or pneumonia, eosinophilic cellulitis (e.g., Wells' syndrome), eosinophilic pneumonia (e.g., Loeffler's syndrome), group, chronic eosinophilic pneumonia), eosinophilic fasciitis (e.g., Schulman's syndrome), delayed hypersensitivity, nonallergic asthma; exercise-induced bronchoconstriction; chronic obstructive pulmonary disease (COPD), acute bronchitis, chronic bronchitis, cough, emphysema; systemic anaphylaxis or hypersensitivity reactions, drug allergies (e.g., to penicillin, cephalosporins), eosinophilia-myalgia syndrome due to ingestion of contaminated tryptophan, insect bite allergy; autoimmune diseases, e.g., rheumatoid arthritis , Graves' disease, Sjogren's syndrome, psoriatic arthritis, multiple sclerosis, systemic lupus erythematosus, myasthenia gravis, immune thrombocytopenia (adult ITP, neonatal thrombocytopenia, pediatric ITP), immune hemolytic anemia (autoimmune and drug-induced), Evans syndrome (platelet and red blood cell immune cytopenia), neonatal Rh disease, Goodpasture's syndrome (anti-GBM disease), celiac disease, autoimmune cardiomyopathy, juvenile onset diabetes mellitus; glomerulonephritis, autoimmune thyroiditis, Behce's disease transplant rejection (e.g. in transplant surgery), including allograft rejection or graft-versus-host disease; inflammatory bowel diseases, such as Crohn's disease and ulcerative colitis; spondyloarthropathy; scleroderma; psoriasis (including T-cell mediated psoriasis) and inflammatory skin diseases, such as dermatitis, eczema, atopic dermatitis, allergic contact dermatitis, urticaria; vasculitis (e.g. necrotizing, cutaneous and hypersensitivity vasculitis); erythema nodosum; eosinophilic myositis, eosinophilic fasciitis, cancer with leukocyte infiltration of the skin or organs;Ophthalmic diseases, such as age-related macular degeneration, diabetic retinopathy and diabetic macular edema, keratitis, eosinophilic keratitis, keratoconjunctivitis, vernal conjunctivitis, scars, anterior eye scars, blepharitis, blepharoconjunctivitis, bullous disorders, cicatricial pemphigoid, conjunctival melanoma, papillary conjunctivitis, dry eye, episcleritis, glaucoma, gliosis, granuloma annulare, Graves' ophthalmopathy, intraocular melanoma, pinguecula, proliferative vitreoretinopathy, pterygium, scleritis, uveitis, acute gout flare, gout or osteoarthritis;

[0058] (5) Pain, for example, chronic idiopathic pain syndromes, neuropathic pain, paresthesia, allodynia, migraine, dental pain and post-operative pain. (6) Depression, anxiety, diabetic neuropathy and bladder disorders, e.g., bladder outlet obstruction, overactive bladder, cystitis; myocardial reperfusion injury or cerebral ischemic injury. In a further aspect, the present invention relates to compounds of general formula 1 for use in the treatment and / or prevention of the above mentioned diseases and conditions. In a further aspect, the present invention relates to the use of compounds of general formula 1 for the preparation of a medicament for the treatment and / or prevention of the above mentioned diseases and conditions. In a further aspect the present invention relates to a method for the treatment or prevention of the above mentioned diseases and conditions, comprising the administration of an effective amount of a compound of general formula 1 to a human.

[0059] Combination therapy The compound 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 selected from the group of therapeutic agents useful in the treatment of the above-mentioned diseases or conditions, in particular fibrotic diseases, inflammatory and immunoregulatory disorders, respiratory or gastrointestinal diseases or complaints, inflammatory diseases of the joints or nasopharynx, eyes and skin, or conditions such as cough, idiopathic pulmonary fibrosis, other interstitial lung diseases, asthma or allergic diseases, eosinophilic diseases, chronic obstructive pulmonary disease, atopic dermatitis, and autoimmune pathologies, for example rheumatoid arthritis and atherosclerosis, or therapeutic agents useful in the treatment of ophthalmological diseases, pain and depression.

[0060] Additional therapeutic agents suitable for such combinations include in particular those which enhance the therapeutic effect of one or more active substances for, for example, one of the mentioned indications and / or allow for a reduction in the dosage of one or more active substances. Thus, the compounds of the present invention may be combined with one or more additional therapeutic agents selected from the group consisting of antifibrotic agents, antitussives, anti-inflammatory agents, anti-atopic dermatitis agents, analgesics, anticonvulsants, anxiolytics, sedatives, skeletal muscle relaxants or antidepressants. Antifibrotic agents include, for example, nintedanib, pirfenidone, phosphodiesterase-IV (PDE4) inhibitors, such as roflumilast, autotaxin inhibitors, such as GLPG-1690 or BBT-877; connective tissue growth factor (CTGF) blocking antibodies, such as pamrevlumab; B-cell activating factor receptor (BAFF-R) blocking antibodies, such as lanalumab; alpha-V / beta-6 blocking inhibitors, such as BG-00011 / STX-100, recombinant pentraxin-2 (PTX-2), such as PRM-151; c-Jun N-terminal kinase (JNK) inhibitors, such as CC-90001; galectin-3 inhibitors, such as TD-139; G protein-coupled receptor 84 (GPR84) inhibitors, such as GLPG-1205; G protein-coupled receptor 84 / G protein-coupled receptor 40 dual inhibitors, such as PBI-4050; Rho-associated coiled-coil-containing protein kinase 2 (ROCK2) inhibitors, such as KD-025; heat shock protein 47 (HSP47) small interfering RNA, such as BMS-986263 / ND-L02-s0201; Wnt pathway inhibitors, such as SM-04646; LD4 / PDE3 / 4 inhibitors, such as tipelukast; recombinant immunomodulatory domain of histidyl-tRNA synthetase (HARS), such as ATYR-1923; prostacyclin, erythropoietin ... staglandin synthase inhibitors, such as ZL-2102 / SAR-191801; 15-hydroxy-eicosapentaenoic acid (15-HEPE, e.g. DS-102); lysyl oxidase-like 2 (LOXL2) inhibitors, such as PAT-1251, PXS-5382 / PXS-5338; phosphoinositide 3-kinase (PI3K) / mammalian target of rapamycin (mTOR) dual inhibitors, such as HEC-68498; calpain inhibitors, such as BLD-2660; mitogen-activated protein kinase kinase kinase (MAP3K19) inhibitors, such as MG-S-2525; chitinase inhibitors, such as OATD-01; mitogen-activated protein kinase-activated protein kinase 2 (MAPKAPK2) inhibitors, such as MMI-0100;Transforming growth factor beta 1 (TGF-beta 1) small interfering RNA, such as TRK250 / BNC-1021; or lysophosphatidic acid receptor antagonists, such as BMS-986278;

[0061] Antitussives are, for example, purinergic receptor 3 (P2X3) receptor antagonists, such as gefapixant, S-600918, BAY-1817080 or BLU-5937; neurokinin 1 (NK-1) receptor antagonists, such as orbepitant, aprepitant; nicotinic acetylcholine receptor alpha 7 subunit agonists, such as ATA-101 / bradanicline; codeine, gabapentin, pregablin or azithromycin.

[0062] Anti-inflammatory agents are, for example, corticosteroids such as prednisolone or dexamethasone; cyclooxygenase-2 (COX2) inhibitors such as celecoxib, rofecoxib, parecoxib, valdecoxib, deracoxib, etoricoxib or lumiracoxib; prostaglandin E2 antagonists; leukotriene B4 antagonists; leukotriene D4 antagonists such as monteleukast; 5-lipoxygenase inhibitors; or other non-steroidal anti-inflammatory agents (NSAIDs) such as aspirin, diclofenac, diflunisal, etodolac, ibuprofen or indomethacin. Anti-atopic dermatitis agents are, for example, cyclosporine, methotrexate, mycophenolate mofetil, azathioprine, phosphodiesterase inhibitors (e.g., apremilast, crisaborole), Janus-associated kinase (JAK) inhibitors (e.g., tofacitinib), IL-4 / IL-13 (e.g., dupilumab), IL-13 (e.g., lebrikizumab, tralokinumab), and neutralizing antibodies against IL-31 (nemolizumab).

[0063] The analgesic may be, for example, of the opioid type, such as morphine, oxymorphine, levopanol, oxycodon, propoxyphene, nalmefene, fentanyl, hydrocodone, hydromorphone, melipidine, methadone, nalorphine, naloxone, naltrexone, buprenorphine, butorphanol, nalbuphine, pentazocine; or of the non-opioid type, such as acetophenamine. Antidepressants include, for example, tricyclic antidepressants, such as amitriptyline, clomipramine, despramine, doxepin, desipramine, imipramine, nortriptyline; selective serotonin reuptake inhibitor antidepressants (SSRIs), such as fluoxetine, paroxetine, sertraline, citalopram, escitalopram; norepinephrine reuptake inhibitor antidepressants (SNRIs), such as maprotiline, lofepramine, mirtazapine, oxaprotiline, fezolamine, tomoxetine, mianserin, bupropion. (buproprion), hydroxybupropion, nomifensine, viloxazine; dual serotonin-norepinephrine reuptake inhibitor antidepressants (SNRIs), such as duloxetine, venlafaxine, desvenlafaxine, levomilnacipran; atypical antidepressants, such as trazodone, mirtazapine, vortioxetine, vilazodone, bupropion; or monoamine oxidase inhibitor antidepressants (MAOIs), such as tranylcypromine, phenelzine or isocarboxazid.

[0064] The anti-anxiety drugs are, for example, benzodiazepines, such as alprazolam, bromazepam, chlordiazepoxide, clonazepam, clorazepate, diazepam, flurazepam, lorazepam, oxazepam, temazepam, triazolam or tofisopam; or they are non-benzodiazepine hypnotics, such as eszopiclone, zaleplon, zolpidem or zopiclone; or they are carbamates, such as meprobamate, carisoprodol, tibamate or lorbamate; or they are antihistamines, such as hydroxyzine, chlorpheniramine or diphenhydramine. The sedatives may, for example, be barbiturates, such as amobarbital, aprobarbital, butabarbital, butabital, mephobarbital, metharbital, methohexital, pentobarbital, secobarbital, talbutal, thiamylal or thiopental; or they may be non-barbiturates, such as glutethimide, meprobamate, methaqualone or dichloralphenazone. Skeletal muscle relaxants are, for example, baclofen, meprobamate, carisoprodol, cyclobenzaprine, metaxalone, methocarbamol, tizanidine, chlorzoxazone or orphenadrine.

[0065] Other suitable combination partners are inhibitors of acetylcholinesterase inhibitors, such as donepezil; 5-HT-3 anatgonists, such as ondansetron; metabotropic glutamate receptor antagonists; antiarrhythmics, such as mexiletine or phenytoin; or NMDA receptor antagonists. Further suitable combination partners are incontinence drugs, for example anticholinergic drugs, such as oxybutynin, tolterodine, darifenacin, fesoterodine, solifenacin or tropium; or they are bladder muscle relaxants, such as mirabegron; or they are alpha blockers, such as tamsulosin, alfuzosin, silodosin, doxazosin or terazosin.

[0066] The dosages of the combination partners mentioned above are usually between 1 / 5 of the lowest normally recommended dose and 1 / 1 of the normally recommended dose. Thus, in another aspect, the present invention relates to the use of a compound according to the invention in combination with one or more additional therapeutic agents as described above or below for the treatment of a disease or condition that can be influenced or mediated by TRPA1, in particular a disease or condition as described above or below. In a further aspect, the present invention relates to a method for treating a disease or condition in a patient that can be affected by inhibition of TRPA1, comprising administering to a patient in need of such treatment a therapeutically effective amount of a compound of formula (I) or a pharma- ceutically acceptable salt thereof in combination with a therapeutically effective amount of one or more additional therapeutic agents. In a further aspect, the present invention relates to the use of a compound of formula (I) or a pharma- ceutically acceptable salt thereof, in combination with one or more additional therapeutic agents, for the treatment of a disease or condition that can be affected by inhibition of TRPA1 in a patient in need thereof.

[0067] In yet another aspect, the invention relates to a method for the treatment of a disease or condition mediated by TRPA1 activity in a patient, comprising the step of administering to a patient, preferably a human, in need of such treatment a therapeutically effective amount of a compound of the invention in combination with a therapeutically effective amount of one or more additional therapeutic agents as described above or below. The use of the compounds according to the invention in combination with an additional therapeutic agent may occur simultaneously or at staggered times. The compound according to the invention and the one or more additional therapeutic agents may be present both together in one formulation, e.g. a tablet or capsule, or separately in two identical or different formulations, e.g. as a so-called kit of parts. Consequently, in another aspect, the present invention relates to a pharmaceutical composition comprising a compound according to the invention and one or more additional therapeutic agents as described above or below, optionally together with one or more inert carriers and / or diluents. In yet another aspect, the present invention relates to the use of a compound according to the invention in a cough measuring device. Other features and advantages of the present invention will become apparent from the following more detailed examples which illustrate, by way of example, the principles of the invention.

[0068] preparation The compounds according to the present invention and their intermediates can be obtained using synthetic methods known to those skilled in the art and described in the organic synthesis literature.Preferably, the compounds are obtained in a manner similar to the preparation methods described below, particularly in the experimental section.In some cases, the order of carrying out reaction steps can be varied.Variants of reaction methods known to those skilled in the art but not described in detail here may also be used. General processes for preparing compounds according to the invention will be apparent to one skilled in the art upon studying the following schemes. Any functional groups in the starting materials or intermediates may be protected using conventional protecting groups. These protecting groups may be cleaved again at a suitable stage in the reaction sequence using methods well known to those skilled in the art. The compounds according to the present invention are prepared by the synthetic methods described below, in which the substituents of the general formula have the meanings described hereinbefore.These methods are intended as illustrations of the present invention, without limiting the subject matter and the scope of the compounds claimed to these examples.If the preparation of starting compounds is not described, they are commercially available or can be prepared analogously to known compounds or methods described herein.Substances described in the literature are prepared according to published synthetic methods.Abbreviations are as defined in the Examples section.

[0069] Scheme 1: [ka] In Scheme 1, compounds of formula I and II can be synthesized via N-alkylation of the appropriate heteroaromatic intermediate (A) or (B) with chloromethylene-oxadiazole (D) in the presence of a base such as potassium carbonate. Alternatively, carboxylic acid (C) can be activated, for example with CDI, and condensed with dihydroxypropaneimide amide (E) to give compounds of type II.

[0070] Scheme 2: [ka]

[0071] In Scheme 2, the alpha-cyanoketone of substructure (F) is reduced in an enantioselective manner by using a suitable catalytic system using a transition metal complex (e.g., of Ru or Ir) in combination with a chiral ligand (e.g., [(1S,2S)-(-)-2-amino-1,2-diphenylethyl](4-toluenesulfonyl)amide) and a hydrogen source such as formate triethylamine complex. Addition of hydroxylamine to the corresponding cyano-alcohol (G) gives the dihydroxypropaneimidamide (E). Ring closure to the chloromethylene-oxadiazole (D) can be achieved by stirring the reaction mixture with chloroacetyl chloride in the presence of a base such as DIPEA.

[0072] Scheme 3: [ka] In scheme 3, 4,5-dichloro-2H-pyridazin-3-one can be N-alkylated with a suitable protecting group such as benzyloxymethyl chloride (Bom-Cl) in the presence of a base such as DBU. Substition of (H) with nitrite at 4-Cl and with hydroxide at 5-Cl position gives intermediate (J), which can be further reacted with ammonia to give (K). Reduction of the nitro group under hydrogen atmosphere using a suitable catalyst such as PtO2 gives di-amine (L), which can be further reacted with carbonyldiimidazole (CDI) to give (M). Finally, di-alkylation with a methylating reagent such as methyl iodide in the presence of a base such as potassium carbonate and cleavage of the protecting group gives intermediate (A).

[0073] Scheme 4: [ka] In Scheme 4, imidazole (O) can be alkylated with a suitable methylating reagent such as methyl iodide (MeI) in the presence of a base such as potassium carbonate. Intermediate (P) can be reacted with an aldehyde (R) via reduction, for example with di-isobutylaluminum hydride (DIBAL-H) ​​and subsequent oxidation, for example with manganese oxide (IV). Condensation of (R) with hydrazine under acidic conditions gives (S), which can be further reduced to intermediate (B). Finally, alkylation of (B) with tert-butyl bromoacetate in the presence of a base such as potassium carbonate gives the ester (T), which can be hydrolyzed under acidic conditions to give (C). EXAMPLES

[0074] preparation The compounds according to the invention and their intermediates can be obtained using synthetic methods known to those skilled in the art and described in the organic synthesis literature, for example, using the methods described in "Comprehensive Organic Transformations", 2nd Edition, Richard C. Larock, John Wiley & Sons, 2010 and "March's Advanced Organic Chemistry", 7th Edition, Michael B. Smith, John Wiley & Sons, 2013. Preferably, the compounds are obtained similarly to the preparation methods as described more fully below, in particular in the experimental section. In some cases, the sequence adopted in carrying out the reaction schemes can be varied. Variants of these reactions known to those skilled in the art but not described in detail here may be used. The general process for preparing the compounds according to the invention will be clear to the skilled person studying the schemes that follow. The starting compounds are commercially available or can be prepared by methods described in the literature or herein, or can be prepared in a similar or similar manner. Any corresponding functional groups in the starting compounds may be protected using conventional protecting groups before the reaction is carried out. These protecting groups may be cleaved again at a suitable stage in the reaction sequence using methods well known to those skilled in the art and described in the literature, e.g., in “Protecting Groups”, 3rd Edition, Philip J. Kocienski, Thieme, 2005 and “Protective Groups in Organic Synthesis”, 4th Edition, Peter GM Wuts, Theodora W. Greene, John Wiley & Sons, 2006. The terms “ambient temperature” and “room temperature” are used interchangeably and designate a temperature of about 20° C., e.g., between 19 and 24° C.

[0075] Abbreviation: [Table 10-1] [Table 10-2]

[0076] Preparation of intermediates Intermediate I Intermediate I.1 (General Route) (3S)-3-(4-chlorophenyl)-3-hydroxypropanenitrile

[0077] [ka] 10.0 g (55.7 mmol) of 4-chlorobenzoylacetonitrile is added to 100 mL of ACN under an inert atmosphere. 142 mg (0.23 mmol) of chloro([(1S,2S)-2-amino-1,2-diphenylethyl](4-toluenesulfonyl)amido)(mesitylene)ruthenium(II) (CAS 174813-81-1) is added, followed by dropwise addition of 8.30 mL (19.8 mmol) of formate triethylamine complex (5:2). After stirring at room temperature for 3 hours, the solvent is removed in vacuo. Water is added to the remaining crude mixture, and the mixture is extracted twice with EtOAc. The organic layers are combined, dried over MgSO4, filtered, and the solvent is removed in vacuo to give intermediate I.1. C9H8ClNO (M=181.6g / mol) ESI-MS: 226[M+HCOO] - R t (HPLC): 0.81 min (Method B)

[0078] The following compounds are prepared using procedures analogous to those described for intermediate I.1, using the appropriate starting materials. As will be appreciated by those skilled in the art, these analogies may involve variations in the general reaction conditions. [Table 11-1] [Table 11-2]

[0079] Intermediate II Intermediate II.1 (General Route) (3S)-3-(4-chlorophenyl)-N,3-dihydroxypropanimidamidl

[0080] [ka] To 9.82 g (54.1 mmol) of (3S)-3-(4-chlorophenyl)-3-hydroxypropanenitrile (Intermediate I.1) in 100 mL of MeOH, 8.00 mL (136 mmol) of hydroxylamine (50% in water) is added and the mixture is stirred for 1.5 h at 75° C. After cooling to room temperature, all volatiles are removed in vacuo to give the crude product, which is used without further purification. C9H 11 ClN2O2 (M = 214.6 g / mol) ESI-MS: 215[M+H] + R t (HPLC): 0.60 min (Method B)

[0081] The following compounds are prepared using procedures analogous to those described for intermediate II.1, using the appropriate starting materials. As will be appreciated by those skilled in the art, these analogies may involve variations in the general reaction conditions.

[0082] [Table 12]

[0083] Intermediate III Intermediate III.1 (General Route) (1S)-2-[5-(chloromethyl)-1,2,4-oxadiazol-3-yl]-1-(4-chlorophenyl)ethan-1-ol

[0084] [ka] To 11.2 g (52.4 mmol) of intermediate II.1 in 55 mL of NMP is added 10.0 mL (57.8 mmol) of DIPEA. After cooling the mixture to 0° C., 4.60 mL (57.7 mmol) of chloroacetyl chloride dissolved in 5 mL of NMP is slowly added and the mixture is stirred at 0° C. for 45 min. Then the mixture is heated to 95° C. and stirring is continued for 4 h. After cooling to room temperature, 200 mL of water is added and the resulting mixture is extracted three times with EtOAc. The organic layers are combined, dried over MgSO4, filtered and the solvent is removed in vacuo. The residue is purified by column chromatography (silica gel; PE / EtOAc, 7 / 3). C 11 H 10 Cl2N2O2 (M = 273.1 g / mol) ESI-MS: 271[MH] - R t (HPLC): 0.93 min (Method B)

[0085] The following compounds are prepared using procedures analogous to those described for intermediate III.1, using the appropriate starting materials. As will be appreciated by those skilled in the art, these analogies may involve variations in the general reaction conditions. [Table 13]

[0086] Intermediate IV Intermediate IV.1 (General Route) 3-(6-fluoro-1-benzothiophen-2-yl)-3-oxopropanenitrile [ka] To 0.63 g (3.00 mmol) of methyl 6-fluoro-1-benzothiophene-2-carboxylate in 9.0 mL of dry toluene and 0.78 mL of dry ACN, 0.36 g (9.00 mmol) of NaH (60% in oil) is added at room temperature under an inert atmosphere. The mixture is heated to reflux, stirred for 16 h, cooled to room temperature, poured into ice / water (30 mL) and treated with 2 M HCl to reach pH=1. EtOAc (20 mL) is added and the phases are separated. The aqueous phase is extracted once more with EtOAc (20 mL), the combined organic phase is washed with brine (20 mL) and the solvent is removed under reduced pressure. The crude product is purified by silica gel column chromatography using a gradient of EtOAc / hexanes (30%-40%). C 11 H6FNOS (M=219.23g / mol) ESI-MS: 218[MH] - R t (HPLC): 3.31 minutes (L) The following compounds are prepared using procedures analogous to those described for intermediate IV.1, using the appropriate starting materials. As will be appreciated by those skilled in the art, these analogies may involve variations in the general reaction conditions.

[0087] [Table 14]

[0088] Intermediate V Intermediate V.1 2-[(Benzyloxy)methyl]-4,5-dichloro-2,3-dihydropyridazin-3-one [ka] To a stirred mixture of 300 g (1.82 mol) of 4,5-dichloro-2H-pyridazin-3-one in 2 L of DMF is added 262 g (1.72 mol) of 1,8-diazabicycloundec-7-ene followed by 370 g (2.37 mol) of chloromethoxymethyl-benzene at 0° C. The reaction mixture is allowed to warm to ambient temperature and stirred for a total of 16 h. The mixture is diluted with water and extracted with ethyl acetate (3×). The combined organic extracts are dried over Na2SO4 and concentrated. The residue is purified by column chromatography (SiO2, EtOAc / PE) to give the desired product. C 12 H 11 N3O5 (M = 285.1 g / mol) ESI-MS: 285 / 287[M+H] + R t (HPLC): 2.2 minutes (K)

[0089] Intermediate V.2 2-[(Benzyloxy)methyl]-4-hydroxy-5-nitro-2,3-dihydropyridazin-3-one

[0090] [ka] To 85.0 g (298 mmol) of intermediate V.1 in 700 mL of DMF and 300 mL of water, 82.3 g (1.19 mol) of sodium nitrite is added. After the reaction mixture is stirred at 85° C. for 24 hours, it is cooled to ambient temperature and acidified with aqueous hydrochloric acid (5N) to pH 4. The mixture is filtered and the collected solid is dried under vacuum to give the desired product. C 12 H 11 N3O5 (M = 277.2 g / mol) ESI-MS: 278[M+H] + R t (HPLC): 1.71 min (Method K)

[0091] Intermediate V.3 4-Amino-2-[(benzyloxy)methyl]-5-nitro-2,3-dihydropyridazin-3-one [ka] To 50.0 g (180 mmol) of intermediate V.2 is added a saturated solution of ammonia in 3.75 L of MeOH. The reaction mixture is heated to 130° C. overnight in an autoclave reactor. After cooling to ambient temperature, the mixture is further cooled to 0° C. The precipitated solid is collected by filtration and purified by column chromatography (SiO2, DCM / MeOH gradient) to give the desired product. C 12 H 12 N4O4 (M = 276.2 g / mol) ESI-MS: 275[MH] - R t (HPLC): 1.76 min (Method J)

[0092] Intermediate V.4 4,5-Diamino-2-[(benzyloxy)methyl]-2,3-dihydropyridazin-3-one [ka] To a stirred mixture of 30.0 g (109 mmol) of intermediate V.3 in 3650 mL of EtOH / EtOAc (4:1) is added 2.47 g (10.9 mmol) of platinum(IV) oxide. The mixture is degassed by passing Ar through the solvent for 15 min. A hydrogen atmosphere (1 atm) is applied and the reaction mixture is stirred for 16 h. After removing the hydrogen atmosphere, the mixture is filtered through a pad of Celite and the filtrate is concentrated to give the desired compound. C 12 H 14 N4O2 (M = 246.3 g / mol) ESI-MS: 247[M+H] + R t (HPLC): 1.62 minutes (Method K)

[0093] Intermediate V.5 5-[(Benzyloxy)methyl]-1H,2H,3H,4H,5H-imidazo[4,5-d]pyridazine-2,4-dione [ka] To a stirred mixture of 29.0 g (118 mmol) of intermediate V.4 in 500 mL of THF, 38.2 g (236 mmol) of 1,1'-carbonyldiimidazole is added and the mixture is stirred at reflux for 16 h. After cooling to ambient temperature, the precipitate is collected by filtration through a sintered funnel. The collected solid is further washed with THF and dried under vacuum to give the desired product. C 13 H 12 N4O3 (M = 272.3 g / mol) ESI-MS: 273[M+H] + R t (HPLC): 1.31 min (Method J)

[0094] Intermediate V.6 5-[(Benzyloxy)methyl]-1,3-dimethyl-1H,2H,3H,4H,5H-imidazo[4,5-d]pyridazine-2,4-dione [ka] To a stirred solution of 28.0 g (103 mmol) of intermediate V.5 in 250 mL of DMF, 42.6 g (309 mmol) of potassium carbonate is added at room temperature and the mixture is stirred for 30 min. It is then cooled to 0° C. and 58.4 g (411 mmol) of methyl iodide is added. After warming to ambient temperature, the reaction mixture is stirred for 16 h. It is quenched by the addition of water and extracted with ethyl acetate (3×). The combined organic extracts are washed with cold water to remove DMF, dried over Na2SO4 and concentrated. The residue is washed with EtOAc / petroleum ether (1:1) to give the desired compound. C 15 H 16 N4O3 (M = 300.3 g / mol) ESI-MS: 301[M+H]+ R t (HPLC): 1.78 min (Method K) 1 H NMR (400 MHz, DMSO-d6) δ ppm 3.37 (s, 3H), 3.55 (s, 3H), 4.63 (s, 2H), 5.52 (s, 2H), 7.25 - 7.34 (m, 5H), 8.35 (s, 1H).

[0095] Intermediate V.7 1,3-Dimethyl-1H,2H,3H,4H,5H-imidazo[4,5-d]pyridazine-2,4-dione

[0096] [ka] A mixture of 20.0 g (66.6 mmol) of intermediate V.6 in 500 mL of DCM is prepared and cooled to −78° C. A solution of 58.4 g (499 mmol) of boron trichloride in DCM is added and the reaction mixture is stirred at the same temperature for 2 h. Then, 500 mL of an equimolar mixture of methanol / DCM is added and the reaction mixture is stirred at −78° C. for another 2 h. After warming to ambient temperature, the mixture is stirred for another 30 min. The mixture is concentrated and coevaporated with MeOH (5 times) to remove traces of methyl boronate. The residue is triturated with MTBE and the collected solid is dried under vacuum to give the desired compound. C7H8N4O2 (M = 180.2 g / mol) ESI-MS: 181[M+H] + R t (HPLC): 0.75 min (Method K) 1 H NMR (500 MHz, DMSO-d6) δ ppm 3.36 (s, 3H), 3.52 (s, 3H), 8.25 (s, 1H), 12.94 (s, 1H).

[0097] Intermediate VI Intermediate VI.1 4,5-Dimethyl 2-bromo-1-methylimidazole-4,5-dicarboxylate [ka] To 30.0 g (114 mmol) of 4,5-dimethyl 2-bromo-1H-imidazole-4,5-dicarboxylate (CAS: 705280-65-5) in 300 mL of DMF, 32.4 g (228 mmol) of iodomethane and 31.5 g (228 mmol) of potassium carbonate are added at ambient temperature. The resulting reaction mixture is stirred at ambient temperature for 16 hours. The mixture is then concentrated, diluted with water and extracted with ethyl acetate. The organic extract is dried over sodium sulfate and concentrated. The residue is purified by column chromatography (silica gel, PE / EtOAc 30 / 70) to give the desired product. C8H9BrN2O4 (M = 277.1 g / mol) ESI-MS: 277 / 279[M+H] + R t (HPLC): 1.64 minutes (Method K)

[0098] Intermediate VI.2 Methyl 2-bromo-5-formyl-3-methylimidazole-4-carboxylate [ka] A mixture of 20.0 g (72.2 mmol) of 4,5-dimethyl 2-bromo-1-methylimidazole-4,5-dicarboxylate in 800 mL of THF is cooled to −78° C. and a solution of 121 mL (180.5 mmol, 25% by weight in toluene) of DIBAL-H is added. The reaction mixture is stirred for 2 hours and allowed to warm slowly to −20° C. during this period. The reaction mixture is then quenched with ice-cold water and a solution of potassium sodium tartrate. The mixture is then extracted with ethyl acetate. The combined organic extracts are washed with saturated brine, dried over sodium sulfate and concentrated. The residue contains the intermediate alcohol in 85% purity and is used in the next step without further purification.

[0099] To a mixture of 15.0 g (85% purity, 51.2 mmol) of ethyl 2-bromo-5-(hydroxymethyl)-3-methylimidazole-4-carboxylate in 150 mL of dichloromethane, add 178 g (2.05 mol) of manganese(IV) oxide at ambient temperature. The resulting suspension is stirred at ambient temperature for 16 hours. The reaction mixture is then filtered through Celite. The filtrate is concentrated under reduced pressure to give the desired product. C7H7BrN2O3 (M = 247.1 g / mol) ESI-MS: 247 / 249[M+H] + R t (HPLC): 1.51 min (Method K)

[0100] Intermediate VI.3 2-Bromo-3-methyl-5H-imidazo[4,5-d]pyridazin-4-one [ka] To a mixture of 8.0 g (32.4 mmol) of methyl 2-bromo-5-formyl-3-methylimidazole-4-carboxylate in 80 mL of ethanol, 1.95 mL (38.9 mmol) of hydrazine hydrate is added at ambient temperature. The reaction mixture is stirred at ambient temperature for 15 minutes, and then 7.4 mL (130 mmol) of acetic acid is added. The reaction mixture is then heated to 80° C. and stirred at this temperature for 30 minutes. After cooling to ambient temperature, the precipitated solid is collected by filtration. It is further washed multiple times with ether and dried under vacuum to obtain the desired product. C6H5BrNO (M=229.0g / mol) ESI-MS: 229 / 231[M+H] + R t (HPLC): 1.34 min (Method K)

[0101] Intermediate VI.4 1-Methyl-1H,6H,7H-imidazo[4,5-d]pyridazin-7-one hydrobromide [ka] To a mixture of 5.3 g (23.1 mmol) of 2-bromo-1-methyl-1H,6H,7H-imidazo[4,5-d]pyridazin-7-one in 75 mL of MeOH, 200 mg of Pd / C (10%) is added in an autoclave reactor. The space above the reaction mixture is evacuated and filled with 50 psi of hydrogen pressure. The reaction mixture is stirred overnight at ambient temperature under 50 psi of hydrogen pressure. After removing the hydrogen and refilling with nitrogen, the reaction mixture is diluted with 300 mL of warm water and filtered. The filtrate is concentrated to dryness to give the desired product. C6H6N4O*HBr (M=231.1g / mol) ESI-MS: 151 [M-HBr+H] + R t (HPLC): 0.12 min (Method C)

[0102] Intermediate VI.5 tert-Butyl 2-{1-methyl-7-oxo-1H,6H,7H-imidazo[4,5-d]pyridazin-6-yl}acetate [ka] To 2.00 g (8.66 mmol) of intermediate VI.4 in 25 mL of DMF, 2.99 g (21.6 mmol) of K2CO3 is added and the mixture is stirred at room temperature for 10 min. 1.90 mL (12.98 mmol) of tert-butyl bromoacetate is added and stirring at room temperature is continued for 4 h. The reaction mixture is poured into 50 mL of water and extracted with EtOAc. The organic layer is treated with MgSO4 and charcoal, filtered and concentrated under reduced pressure. The crude material is stirred with isopropanol, filtered and the collected precipitate is dried at 50 °C to give the desired product. C 12 H 16 N4O3 (M = 264.28 g / mol) ESI-MS: 265[M+H] + R t(HPLC): 0.82 min (Method B)

[0103] Intermediate VI.6 2-{1-methyl-7-oxo-1H,6H,7H-imidazo[4,5-d]pyridazin-6-yl}acetic acid hydrochloride [ka]

[0104] To 1.0 g (3.78 mmol) of intermediate VI.5 in 10 mL of dioxane is added 20 mL of HCl in dioxane (4 mol / L) and the reaction mixture is stirred at room temperature for 3 days. The precipitate is filtered off, washed with isopropanol and dried at 50° C. to give the desired product. C8H8N4O3*HCl (M=244.6g / mol) ESI-MS: 209 [M-HCl+H] + R t (HPLC): 0.42 min (Method B)

[0105] Preparation of final compounds Example 1 (General Procedure) 5-({3-[(2S)-2-(4-chlorophenyl)-2-hydroxyethyl]-1,2,4-oxadiazol-5-yl}methyl)-1,3-dimethyl-1H,2H,3H,4H,5H-imidazo[4,5-d]pyridazine-2,4-dione

[0106] [ka] To 200 mg (1.11 mmol) of intermediate V.7 in 5 ml of DMF is added 380 mg (2.75 mmol) of K2CO3, and the mixture is stirred at room temperature for 10 min. Then, 250 mg (0.92 mmol) of intermediate III.1 in 5 mL of DMF is added, and stirring is continued overnight. The reaction mixture is filtered, and the filtrate is purified by reverse phase HPLC (ACN / H2O gradient, 0.1% TFA) to give the desired product. C18 H 17 ClN6O4 (M = 416.8 g / mol) ESI-MS: 417[M+H] + R t (HPLC): 0.86 min (Method B) 1 H NMR (400 MHz, DMSO-d6) δ ppm: 2.92 - 3.05 (m, 2 H), 3.39 (s, 3 H), 3.53 (s, 3 H), 4.92 - 4.99 (m, 1 H), 5.63 - 5.64 (m, 2 H), 7.32 - 7.37 (m, 4 H), 8.40 (s, 1 H).

[0107] The following compounds are prepared using the appropriate starting materials using procedures analogous to those described for the general procedure in Example 1. As will be appreciated by those skilled in the art, these analogous examples may involve variations in the general reaction conditions. [Table 15-1] [Table 15-2] Analytical data for the compounds described in the table below:

[0108] [Table 16-1] [Table 16-2]

[0109] Example 8 6-({3-[(2S)-2-(4-bromophenyl)-2-hydroxyethyl]-1,2,4-oxadiazol-5-yl}methyl)-1-methyl-1H,6H,7H-imidazo[4,5-d]pyridazin-7-one

[0110] [ka] To 73 mg (0.30 mmol) of intermediate VI.6 in 3 ml of DMF, 52 μL (0.30 mmol) of DIPEA and 58 mg (0.36 mmol) of CDI are added and the mixture is stirred at room temperature for 2 min. Then, 78 mg (0.30 mmol) of intermediate II.6 is added and stirring is continued at 100° C. for 4 h. After cooling to ambient temperature, the reaction mixture is purified by reverse phase HPLC (ACN / H2O gradient, 0.1% TFA) to give the desired product. C 17 H1BrlN6O3 (M = 431.2 g / mol) ESI-MS: 431 / 433[M+H] + R t (HPLC): 0.86 min (Method B) 1 H NMR (400 MHz, DMSO-d6) δ ppm: 2.90 - 3.05 (m, 2 H), 4.05 (s, 3 H), 4.94 (dd, J=7.8, 5.8 Hz, 1 H), 5.65 (s, 2 H), 7.26 - 7.32 (m, 2 H), 7.44 - 7.50 (m, 2 H), 8.40 (s, 1 H), 8.50 (s, 1 H)

[0111] Analytical HPLC method Method A [Table 17]

[0112] Method B [Table 18]

[0113] Method C [Table 19]

[0114] Method D

Table 20

[0115] Method E

Table 21

[0116] Method F

Table 22

[0117] Method G

Table 23

[0118] Method H

Table 24

[0119] Method I

Table 25

[0120] Method J

Table 26

[0121] Method K

Table 27

[0122] Method L

Table 28

[0123] Method M

Table 29

[0124] Method N

Table 30

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[10] The compound of formula (I) according to any one of the above [1] to [7], or a pharma- ceutically acceptable salt thereof, for use as a medicine.

[11] The compound according to any one of the above [1] to [7], or a pharma- ceutically acceptable salt thereof, for treating or preventing an inflammatory airway disease, a fibrotic disease, or cough.

[12] The compound according to any one of the above [1] to [7], or a pharma- ceutically acceptable salt thereof, for treating or preventing idiopathic pulmonary disease (IPF) or cough.

Claims

1. A compound of formula (I). [Formula 1] (I) (In the formula, (1) A is selected from the group consisting of phenyl, thiophenyl, benzothiophenyl, and benzofuranyl, where A is unsubstituted or selected from the group consisting of halogen and C 1-4 -alkyl radical R 1 or is replaced by one or two members of Or, (2) A is a group of the formula: [Case 2] and E is a group of the formula: [C3] (selected from the group consisting of

2. R 1 is Br, Cl, F and H 3 2. The compound of claim 1, selected from the group consisting of C.

3. (1) A is a group of the following chemical formula: [C4] where A is unsubstituted or a group R 1 or is replaced by one or two members of Or, (2) A is a group of the formula: [C5] 2. The compound of claim 1,

4. A is a group of the formula: [chemical 1-1] [C6-2] 2. The compound of claim 1 selected from the group consisting of:

5. The compound of formula (I) is a compound of the following formula: [C7] 2. The compound of claim 1 , selected from:

6. The compound of formula (I) is a compound of the following formula: [C8] 2. The compound of claim 1 , selected from:

7. The compound of formula (I) is a compound of the following formula: [Case 9-1] [C9-2] [C9-3] 2. The compound of claim 1 selected from the group consisting of:

8. A pharma- ceutically acceptable salt of the compound according to any one of claims 1 to 7.

9. A pharmaceutical composition comprising at least one compound of formula (I) as defined in any one of claims 1 to 7 or a pharma- ceutically acceptable salt thereof, optionally comprising one or more pharma- ceutically acceptable excipients.

10. 10. The pharmaceutical composition according to claim 9 for the treatment or prevention of inflammatory airways diseases or fibrotic diseases or cough.

11. 10. The pharmaceutical composition according to claim 9 for the treatment or prevention of idiopathic pulmonary disease (IPF) or cough.

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