Uracil derivatives as TRPA1 inhibitors

Abstract

The present disclosure provides certain uracil derivatives that are inhibitors of Transient Receptor Potential Ankyrin 1 (TRPA1) and are therefore useful for treating diseases treatable by inhibition of TRPA1, as well as pharmaceutical compositions containing the compounds and processes for preparing the compounds. TIFF2024512972000062.tif45150

Description

[Technical field]

[0001] The present disclosure provides certain uracil derivatives that are inhibitors of transient receptor potential ankyrin 1 (TRPA1) and are therefore useful in the treatment of diseases that can be treated by TRPA1 inhibition, as well as pharmaceutical compositions containing same and methods for preparing said compounds. [Background technology]

[0002] Transient receptor potential channels (TRP channels) are a family of voltage-gated ion channels located primarily in the plasma membrane of many mammalian cell types. There are approximately 30 structurally related TRP channels, which are 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 at the N-terminus of human TRPA1), hence the "A" in the ankyrin name (Montell, 2005). TRPA1 is highly expressed in the plasma membrane of sensory neurons in the dorsal root and nodose ganglia in both the skin and lung, 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 resulting in somatosensory modalities such as pain, cold, and itch. TRPA1 is activated by several reactive electrophilic stimuli (e.g., allyl isothiocyanate, reactive oxygen species), as well as non-reactive compounds (e.g., icilin), and is involved in cough associated with asthma, chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis (IPF), or cough after viral infection, or chronic idiopathic cough, as well as cough in susceptible patients (Song and Chang, 2015; Grace and Belvisi, 2011). TRPA1 inhibitors may be useful in treating IPF, where cough is prevalent, due to the association between cough and lung injury, based on studies showing cough-induced increases in 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 resulting from 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 via consumption of spicy foods is thought to modulate the Nrf2 pathway and reduce oxidative stress (Bousquet et al., 2020, Bousquet et al., 2021). Therefore, inhibitors of TRPA1 have potential for the treatment of Covid-19 / SARS-Cov-2 induced lung injury. TRPA1 antagonists inhibit calcium signaling caused by cough triggers such as cigarette smoke extract (CSE) oxidative stress, release of inflammatory mediators, and downregulation of antioxidant gene expression (Lin et al., 2015; Wang et al., 2019). TRPA1 antagonists are effective in the study of atopic dermatitis (Oh et al., 2013; Wilson et al., 2013), contact dermatitis (Liu et al., 2013), psoriasis-associated itch (Wilson et al., 2013), and IL-31-dependent itch (Cevikbas et al., 2014).Human TRPA1 gain of function is associated with familial episodic pain syndrome (Kremeyer et al., 2010). TRPA1 antagonists were effective in a behavioral model of migraine-related allodynia (Edelmayer et al., 2012). TRPA1 is selectively increased in the trigeminal ganglia innervating injured teeth compared with TRPA1 expression in the trigeminal ganglia innervating healthy teeth (Haas et al., 2011). Several anesthetics, including isoflurane, are known to be TRPA1 agonists (Matta et al., 2008), providing a rationale for TRPA1 inhibitors for postoperative pain relief. TRPA1 knockout and wild-type mice treated with TRPA1 antagonists exhibited anxiolytic- and antidepressant-like phenotypes (de Moura et al., 2014). TRPA1 inhibitors are expected to be beneficial in the treatment of diabetic neuropathy based on studies showing a mechanistic relationship of counterregulation between AMPK and TRPA1 (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). In a monosodium urate mouse gout model, urate crystals and arthritis are reduced in TRPA1 knockout mice (Moilanen et al., 2015). TRPA1 deficiency in rats improved joint inflammation and hyperalgesia in a rat acute gout attack model ( Trevisan et al., 2014 ). Activation of TRPA1 induces an inflammatory response in osteoarthritic chondrocytes ( Nummenmaa et al., 2016 ).Inhibition and genetic deletion of TRPA1 reduces inflammatory mediators in osteoarthritic mouse chondrocytes and mouse cartilage (Nummenmaa et al., 2016). Finally, TRPA1 knockout mice showed improved weight bearing of osteoarthritic limbs in a MIA-induced knee swelling model (Horvath et al., 2016). TRPA1 is differentially expressed in the bladder epithelium of rats (Du et al., 2007) and patients (Du et al., 2008) with bladder outlet obstruction. Modulation of TRPA1 receptors attenuates bladder overactivity in a rat spinal cord injury model (Andrade et al., 2011), and intrathecal administration of a TRPA1 antagonist reduces cyclophosphamide-induced cystitis in hypermicrium-hypermic rats (Chen et al., 2016).

[0004] It would therefore be desirable to provide potent TRPA1 inhibitors. TRPA1 inhibitors of various structural classes are reviewed in S.Skerratt, Progress in Medicinal Chemistry, 2017, Volume 56, 81-115 and 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. WO2017 / 060488 discloses a compound having the general structural formula: [ka] The present invention discloses a compound having the formula: Examples 53, 72, 73, 86 and 90 therein disclose TRPA1 activity, with IC values ​​of less than 100 nM in a calcium flux assay. 50 has.

[0005] L. Schenkel, et al., J. Med. Chem. 2016, 59, 2794-2809, has the general structure [ka] The present invention discloses quinazolinone-based TRPA1 antagonists, including compounds of the formula: 50 It is disclosed as having an antagonistic TRPA1 activity of 58 nM and an intrinsic clearance of <14 μL / min / kg in human liver microsomes. Summary of the Invention

[0006] The present invention discloses novel uracil derivatives that are inhibitors of transient receptor potential ankyrin 1 (TRPA1), possessing suitable pharmacological and pharmacokinetic properties that enable their use as drugs for the treatment of conditions and / or diseases that can be treated by inhibition of TRPA1. The compounds of the present invention may offer a variety of advantages, such as improved efficacy, increased metabolic and / or chemical stability, high selectivity, safety and tolerability, improved solubility, improved membrane permeability, desirable plasma protein binding, improved bioavailability, suitable pharmacokinetic properties, and the potential to form stable salts.

[0007] 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 uracil derivative further characterized by: The compounds of the present invention are structurally distinct from Examples 53, 72, 73, 86, and 90 of WO2017 / 060488 and Example 31 of L. Schenkel, et al., J. Med. Chem. 2016, 59, 2794-2809 in that they contain a substituted uracil core and a substituent adjacent to the secondary aliphatic alcohol. These structural differences unexpectedly result in 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 as an initial screening step in the context of selecting and / or designing drugs with favorable pharmacokinetic properties. The primary site of metabolism for many drugs is the liver. Human liver microsomes contain cytochrome P450 (CYP) and therefore serve as a model system for in vitro drug metabolism phase I studies. Improved stability in human liver microsomes is associated with several advantages, including increased bioavailability and sufficient half-life, which can allow for lower and more frequent administration to patients. Thus, improved stability in human liver microsomes is a favorable feature for a compound to be used as a drug. Thus, in addition to being able to inhibit TRPA1, the compounds of the present invention are expected to have favorable in vivo clearance, thereby having the 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 site of metabolism for many drugs is the liver. Human hepatocytes contain cytochrome P450 (CYP) and other drug metabolizing enzymes and therefore serve as a model system for in vitro drug metabolism studies. (Importantly, in contrast to liver microsomal assays, hepatocyte assays also cover phase II biotransformation as well as liver-specific transporter-mediated processes and therefore serve as a more complete system for drug metabolism studies). Improved stability in human hepatocytes is associated with several advantages, including increased bioavailability and sufficient half-life, which may allow for lower and more frequent administration to patients. Thus, improved stability in human hepatocytes is a favorable feature for compounds used as drugs. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

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

[0010] Another embodiment of the invention is a phenyl-, thiophenyl-, benzofuranyl- and benzothiophenyl-containing aryl group, wherein A is unsubstituted or contains one or two groups R consisting of F, Cl, I and CH3. 1 The present invention relates to compounds of formula (I) which are substituted with Another embodiment of the present invention is where A is selected from the group consisting of phenyl, benzofuranyl and benzothiophenyl, and A is unsubstituted or selected from halogen and C 1-4 - one or two radicals R consisting of alkyl1 The present invention relates to compounds of formula (I) which are substituted with Another embodiment of the invention is a phenyl-, benzofuranyl- and benzothiophenyl-substituted aryl group, wherein A is unsubstituted or contains one or two groups R consisting of F, Cl, I and CH3. 1 The present invention relates to compounds of formula (I) which are substituted with Another embodiment of the present invention is wherein A is [ka] and A is unsubstituted or is selected from the group consisting of one or two groups R 1 and R 1 relates to a compound of formula (I), as defined in any of the previous embodiments.

[0011] The preferred [ka] TIFF0007617306000006.tif39170 TIFF0007617306000007.tif38170 TIFF0007617306000008.tif39170 TIFF0007617306000009.tif39170 TIFF0007617306000010.tif3980 and [ka] A compound according to formula (I) selected from the group consisting of:

[0012] 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 this disclosure and the context, except that, as used herein, unless specified to the contrary, the following terms have the meaning indicated and are subject to the following conventions: 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. In general, for groups such as HO, H2N, (O)S, (O)2S, NC (cyano), HOOC, F3C, etc., one skilled in the art can determine the point of attachment of the radical to the molecule from the free valence of the group itself. In the case of combined 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, 1-3 The alkyl group is attached to a core or group to which the substituents are attached.

[0013] When a compound of the invention is depicted by a chemical name and a chemical formula, the chemical formula shall prevail in the event of a conflict. An asterisk may be used in a sub-formula to indicate a bond that is connected to a defined core molecule. Numbering of substituent atoms begins with the most proximal atom of the core or 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 3-position carbon atom of the propyl group. The term "1-methylpropyl-", "2,2-dimethylpropyl-" or "cyclopropylmethyl-" refers to the following radicals: [ka] Represents.

[0014] An asterisk can be used in a sub-formula to indicate a bond that is connected to a defined core molecule. The term “C 1-n"-alkyl", alone or in combination with another radical, denotes an acyclic, saturated, branched or straight-chain hydrocarbon radical having 1 to n C atoms, where n is an integer selected from 2, 3, 4, or 5. For example, the term C 1-5 -Alkyl includes the radicals H3C-, H3C-CH2-, H3C-CH2-CH2-, H3C-CH(CH3)-, H3C-CH2-CH2-CH2-, H3C-CH2-CH(CH3)-, H3C-CH(CH3)-CH2-, H3C-C(CH3)2-, H3C-CH2-CH2-CH2-CH2-, H3C-CH2-CH2-CH2-, H3C-CH2-CH2-CH(CH3)-, H3C-CH2-CH(CH3)-CH2-, H3C-CH2-CH(CH3)-CH2-, H3C-CH(CH3)-CH2-CH2-, H3C-CH2-C(CH3)2-, H3C-C(CH3)2-CH2-, H3C-CH(CH3)-CH(CH3)-CH(CH3)-, and H3C-CH2-CH(CH2CH3)-. The term "fluoro" appended to an "alkyl," "alkylene," or "cycloalkyl" group (saturated or unsaturated) refers, for example, to an alkyl or cycloalkyl group in which one or more hydrogen atoms have been replaced with a fluorine atom. Examples include, but are not limited to, H2FC-, HF2C-, and F3C-.

[0015] The term phenyl refers to the following ring: [ka] Refers to the radical of The term thiophenyl refers to the following ring: [ka] Refers to the radical of 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 The term uracil is the following core [ka] Refers to the radical of

[0016] As used herein, the term "substituted" means that any one or more hydrogens on the designated atom are replaced with a selection from the indicated group, provided that the normal valence of the designated atom is not exceeded and that the substitution results in a stable compound. Unless otherwise 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.), as well as racemates thereof, as well as mixtures of different ratios of the separate enantiomers, mixtures of diastereomers, or mixtures of any of the aforementioned forms in which such isomers and enantiomers exist, as well as pharmaceutically acceptable salts thereof and solvates thereof, such as, for example, hydrates, examples of solvates include, for example, solvates of the free compound or of a salt of the compound. In general, substantially pure stereoisomers can be obtained according to synthetic principles known to those skilled in the art, for example, by separation of the corresponding mixtures, by the use of stereochemically pure starting materials, and / or by stereoselective synthesis. Methods for preparing optically active forms are known in the art, for example, by resolution of racemates or, for example, by synthesis starting from optically active starting materials and / or by using chiral reagents.

[0017] Enantiomerically pure compounds or intermediates of the present invention may also be prepared via asymmetric synthesis, for example by the preparation and subsequent separation of suitable diastereomeric compounds or intermediates which can be separated by known methods (e.g. by chromatographic separation or crystallization) and / or by the use of chiral reagents, such as chiral starting materials, chiral catalysts or chiral auxiliaries. Furthermore, methods for preparing enantiomerically pure compounds from the corresponding racemic mixture are known to the skilled artisan, for example by chromatographic separation of the corresponding racemic mixture on a chiral stationary phase; or by resolution of the racemic mixture using a suitable resolving agent, for example by formation of diastereomeric salts 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 (for example 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.

[0018] The expression "pharmacologically acceptable" is used herein to refer to compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use without undue toxicity, irritation, allergic response, or other problem or complication, and commensurate with a reasonable benefit / risk ratio. 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 containing 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 include 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 containing 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 the 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 other acids than those mentioned above, useful, for example, for the purification or isolation of the compounds of the invention (eg, trifluoroacetates), also form part of the invention.

[0019] Biological assays Assessment of TRPA1 activity Assay A: TRPA1 assay The activity of the compounds of the present invention can be demonstrated using the following in vitro TRPA1 cellular assay.

[0020] method: The human HEK293 cell line (Perkin Elmer, product number AX-004-PCL) overexpressing the human TRPA1 ion channel is used as a test system for compound efficacy and potency. Compound activity is determined by measuring the effect of the compound on intracellular calcium concentration induced by AITC (allyl isothiocyanate) agonism in the FLIPRtetra system (Molecular Devices). Cell culture: Cells are obtained as frozen cells in cryo-vials and stored at -150°C until use. Cells are grown in culture medium (MEM / EBSS medium with 10% FCS and 0.4 mg / ML Geneticin). It is important that the density does not exceed 90% confluence. For subculture, cells are detached from the flask with Versene. The day before the assay, cells are detached, washed twice with medium (MEM / EBSS medium with 10% FCS) and 20,000 cells are seeded at 20 μl / well into a Corning poly-D-lysine biocoated 384-well plate (black, clear bottom, cat. no. 356697). Plates are incubated for 24 hours at 37°C / 5% CO2 before use in the assay.

[0021] compound preparation Test compounds are dissolved in 100% DMSO at a concentration of 10 mM and diluted in DMSO to a concentration of 5 mM in a first step, followed by serial dilution steps in 100% DMSO. The dilution factor and number of dilution steps can be changed according to needs. Typically, eight concentrations are prepared at 1:5 dilutions, and further intermediate dilutions of the substances (1:20) are performed in HBSS / HEPES buffer (1×HEPES, Gibco, Cat. No. 14065; 20 mM HEPES, SIGMA, Cat. No. 83264; 0.1% BSA, Invitrogen, Cat. No. 11926, pH 7.4). FLIPR Assay: On the day of the assay, cells are washed three times with assay buffer, leaving 20 μL of buffer in the wells after washing. 10 μL of loading buffer in HBSS / HEPES from the Ca6 kit (catalog no. R8191, Molecular Devices) is added to the cells, the plate is covered and incubated for 120 min at 37°C / 5% CO2. 10 μL of compound or control in HBSS / HEPES buffer / 5% DMSO from the intermediate dilution plate is carefully added to the wells. Luminescence (indicating calcium influx or release) is read on the FLIPRtetra instrument for 10 min to monitor the effect of the compound (e.g. agonism). Finally, 10 μL of the agonist AITC at 50 μM dissolved in HBSS / HEPES buffer / 0.05% DMSO (final concentration 10 μM) is added to the wells, after which an additional reading is taken on the FLIPRtetra instrument for 10 min. The area under the signal curve (AUC) after addition of AITC is used to calculate IC50 / % inhibition. Data evaluation and calculations: Each assay microtiter plate contains wells with a vehicle (1% DMSO) control instead of compound as a control for AITC-induced luminescence (100% CTL; high control) and wells with a vehicle control without AITC as a control for non-specific changes in luminescence (0% CTL; low control). The data is analyzed by calculating the area under the signal curve for each individual well. Based on this value, the MegaLab software (developed in-house) is used to calculate the percentage value of the measured value for each substance concentration ((AUC(sample)-AUC(low)). * 100 / (AUC(high)-AUC(low)). IC50 values ​​are calculated from % control values ​​using MegaLab software. Calculation: [y=(ad) / (1+(x / c)^b)+d], where a=low value, d=high value; x=concentration M; c=IC50 M; b=Hill coefficient; y=% control

[0022] [Table 1] [Table 2] [Table 3]

[0023] Assessment of microsomal clearance Assay B: Microsomal Clearance: Pooled liver microsomes are used to assay metabolic degradation of test compounds at 37° C. A final incubation volume of 100 μL at each 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 RT. After a short preincubation at 37°C, the reaction is started by adding beta-nicotinamide adenine dinucleotide phosphate, reduced (NADPH, 1 mM) and terminated by transferring aliquots to the solvent at different time points (0, 5, 15, 30, 60 min). In addition, incubations without NADPH are monitored for NADPH-independent degradation and terminated at the final time point. The remaining percentage [%] of the test compound after NADPH-independent incubation is reflected by the parameter c (control) (metabolic stability). Quenched incubations are 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.

[0024] The intrinsic clearance (CL_INTRINSIC) is calculated taking into account the amount of protein during 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, human: 21 ml / (min x kg)

[0025] [Table 4] [Table 5] [Table 6]

[0026] 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 of preincubation in an incubator (37 °C, 10% CO), add 5 µl of test compound solution (80 µM; dilute 2 mM DMSO stock solution 1:25 in medium) to 395 µl of hepatocyte suspension (cell density ranges from 0.25-5 Mio cells / mL depending on species, typically 1 Mio cell / mL; final concentration of test compound 1 µM, final DMSO concentration 0.05%).

[0027] Cells are incubated for 6 hours (incubator, orbital shaker) and samples (25 μl) are taken at 0, 0.5, 1, 2, 4 and 6 hours. Samples are transferred to 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+cluster / k)×1000 / 60, where C0: initial concentration of incubation [μM], CD: cell density of viable cells [10e6 cells / mL], AUD: area under the data [μM×time], cluster: concentration of last data point [μM], k: slope of regression line of decrease of parent compound [time-1].

[0028] The calculated in vitro hepatic intrinsic clearance can be scaled up to the in vivo hepatic intrinsic clearance by using a liver model (well stirred model), and can be used to predict the in vivo hepatic blood flow clearance (CL). CL_INTRINSIC_INVIVO[ml / min / kg]=(CL_INTRINSIC[μL / min / 10e6 cells] x hepatocellularity [10e6 cells / g liver] x 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]) Hepatocyte cellularity, human: 120 x 10e6 cells / g liver Hepatic factor, human: 25.7 g / kg body weight Blood flow, human: 21 ml / (min x kg)

[0029] [Table 7] [Table 8] [Table 9]

[0030] Permeability assessment Caco-2 cells (cells 1-2 x 10 5 pieces / area 1cm 2) are inoculated 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 (e.g., DMSO, 1-20 mM stock solution). 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 H2O, 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 (three filter replicates), respectively. Samples are collected from the donor at the beginning and end of the experiment, as well as from the receiver side at various time intervals up to 2 hours, for concentration determination by HPLC-MS / MS or scintillation counting. The sampled receiver volume is replaced with fresh receiver solution.

[0031] Assessment of plasma protein binding This parallel dialysis (ED) technique is used to determine the approximate fractional binding of test compounds to plasma proteins in vitro. Dianorm Teflon dialysis cells (0.2 micro) are used. Each cell consists of a donor and an acceptor chamber, separated by an ultrathin semipermeable membrane with a molecular weight cutoff of 5 kDa. Stock solutions for each test compound are prepared in DMSO at 1 mM and diluted to a final concentration of 1.0 μM. Subsequent dialysis solutions are prepared in pooled human or rat plasma (with NaEDTA) from male and female donors. Aliquots of 200 μL of dialysis buffer (100 mM potassium phosphate, pH 7.4) are distributed into the buffer chamber. Aliquots of 200 μL of the dialysis solution of the test compound are distributed into the plasma chamber. Incubation is carried out for 2 hours at 37°C under rotation. At the end of the dialysis time, the dialysate is transferred to a reaction tube. The tube for the buffer fractionation 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 ACN / water (80 / 20), 25 μL buffer, 25 μL calibration solution and 25 μL 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 blank plasma, 25 μL internal standard solution and 200 μL ACN. The samples are measured by the HPLC-MS / MS instrument and evaluated with the Analyst-Software. The binding percentage is calculated by the following formula: % binding = (plasma concentration - buffer concentration / plasma 30 concentration) x 100.

[0032] Solubility evaluation Saturated solutions are prepared in a well plate (configuration dependent on 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 agitated for a predefined time (typically in the range of 2-24 hours) and then filtered using a suitable filter membrane (typically a PTFE-filter with a pore size of 0.45 μm). The first few drops of filtrate are discarded to avoid filter absorption. The amount of dissolved drug substance is determined by UV spectroscopy. Additionally, the pH of the saturated aqueous solution is measured using a glass electrode pH meter.

[0033] Pharmacokinetic characterization Test compounds are administered to each test species either intravenously or orally. At several time points after test compound administration, blood samples are taken, anticoagulated and centrifuged. Analyte concentrations - administered compound 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.

[0034] Assessment of metabolism in human hepatocytes in vitro Human primary hepatocyte suspensions are used to investigate the metabolic pathway of test compounds. After recovery from cryopreservation, 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 final cell density was 1.0 cells. * 10 6 ~4.0 * 10 6 Test compound solutions are mixed into the hepatocyte suspension to obtain cells / ml (depending on the compound turnover rate observed in primary human hepatocytes), a final test compound concentration of 10 μM, and a final DMSO concentration of 0.05%. 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, and resuspended prior to bioanalysis by liquid chromatography-high resolution mass spectrometry and identification of putative metabolites. The structure is Fourier-Transform-MS n Assignments are tentatively made based on the data. Metabolites are reported as percentage of parent in human hepatocyte incubations with a threshold of ≥ 4%.

[0035] Treatment method The present invention is directed to compounds of general formula 1, which are useful for the prevention and / or treatment of diseases and / or conditions associated with or modulated by TRPA1 activity, including, but not limited to, the treatment and / or prevention of fibrotic diseases, inflammatory and immunoregulatory disorders, respiratory or gastrointestinal diseases or complaints, ophthalmic 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 pulmonary interstitial 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 neurological disorders such as pain and acute pain, post-operative pain, chronic pain, and depression, and bladder disorders.

[0036] The compounds of general formula 1 are useful for the prevention and / or treatment of: (1) Cough, including chronic idiopathic cough or chronic refractory cough, cough associated with asthma, COPD, lung cancer, post-viral and idiopathic pulmonary fibrosis, and other pulmonary interstitial diseases. (2) Pulmonary fibrotic diseases such as pneumonia or interstitial pneumonia associated with collagen diseases. Examples of collagen diseases include lupus erythematosus, systemic sclerosis, rheumatoid arthritis, polymyositis and dermatomyositis, idiopathic interstitial pneumonia such as 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 pneumonia due to occupational exposure such as asbestosis, silicosis, miner's lung (coal dust), farmer's lung (hay and mold), pigeon breeder's lung (birds), or interstitial pneumonia due to metal dust or mycobacteria. These include interstitial pneumonias with a clear cause such as interstitial pneumonia due to occupational airborne or other triggers such as pulmonary embolism, or interstitial pneumonia as a result of treatment such as radiation, methotrexate, amiodarone, nitrofurone or chemotherapy, or granulomatous diseases such as granulomatosis with polyangiitis, Churg-Strauss syndrome, sarcoidosis, hypersensitivity pneumonitis, or interstitial pneumonia caused by different causes such as aspiration, inhalation of toxic gases, vapors, bronchitis or pneumonitis or interstitial pneumonia caused by heart failure, x-rays, radiation, chemotherapy, M. boeck or sarcoidosis, granulomatous diseases, cystic fibrosis or pancreatic 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 hepatic bridging fibrosis, liver cirrhosis, nonalcoholic steatohepatitis (NASH), atrial fibrosis, endomyocardial fibrosis, old myocardial infarction, glial scar, atherosclerosis, fibrous arthritis, Dupuytren's contracture, keloid, scleroderma / systemic sclerosis, mediastinal fibrosis, myelofibrosis, Peyronie's disease, nephrogenic systemic fibrosis, retroperitoneal fibrosis, and adhesive arthritis. (4) Inflammatory, autoimmune or allergic diseases and conditions, such as allergic or non-allergic rhinitis or sinusitis, chronic sinusitis or rhinitis, nasal polyps, chronic rhinosinusitis, acute rhinosinusitis, asthma, childhood asthma, allergic bronchitis, alveolitis, hypersensitive airways, allergic conjunctivitis, bronchiectasis, adult respiratory syndrome, bronchial and pulmonary edema, bronchitis or pneumonitis, eosinophilic cellulite (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, etc.; exercise-induced bronchoconstriction; chronic obstructive pulmonary disease. chronic bronchitis, cough, emphysema; systemic anaphylaxis or hypersensitivity reactions, drug allergies (e.g. to penicillins, cephalosporins), eosinophilic polymyalgia syndrome due to ingestion of contaminated tryptophan, insect bite allergies; autoimmune diseases, e.g. rheumatoid arthritis, Graves' disease, Sjogren's syndrome, psoriatic arthritis, multiple sclerosis, systemic lupus erythematosus, gravitational myasthenia, 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 (Rh disease), Goodpasture's syndrome (anti-GBM disease), celiac disease, autoimmune cardiomyopathy, juvenile onset diabetes; glomerulonephritis, autoimmune thyroiditis, Behcet's disease; graft rejection (e.g. at the time of transplantation) including allograft rejection or graft-host disease; inflammatory bowel diseases such as Crohn's disease and ulcerative colitis; spondyloarthropathy; scleroderma; inflammatory skin diseases such as psoriasis (including T-cell psoriasis) and dermatitis, eczema, atopic dermatitis, allergic contact dermatitis, urticaria; vasculitis (e.g. necrotizing, cutaneous and hypersensitivity vasculitis); erythema nodosum; eosinophilic myositis, eosinophilic fasciitis, cancer with infiltration of skin or organs with leukocytes;Ophthalmic diseases such as age-related macular degeneration, diabetic retinopathy and diabetic macular edema, keratitis, eosinophilic keratitis, keratoconjunctivitis, vernal keratoconjunctivitis, eye disorders, anterior eye disorders, blepharitis, blepharoconjunctivitis, bullous disease, cicatricial pemphigoid, conjunctival melanoma, papillary conjunctivitis, dry eye, episcleritis, glaucoma, gliosis, granuloma annulare, Graves' ophthalmopathy, intraocular melanoma, pinguecula, proliferative vitreous retinopathy, pterygium, scleritis, uveitis, acute gout attacks, gout or ocular osteoarthritis; (5) Pain such as chronic idiopathic pain syndrome, neuropathic pain, paresthesia, allodynia, migraine, toothache, and postoperative pain. (6) Depression, anxiety neurosis, diabetic neuropathy, and bladder disorders such as bladder outlet obstruction, overactive bladder, and cystitis; myocardial reperfusion injury or cerebral ischemic injury.

[0037] Thus, the present invention relates to compounds of general formula 1 for use as medicines. 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 which are associated with or modulated by TRPA1 activity. 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, pain and neurological disorders, including inflammatory and immunoregulatory disorders such as cough, idiopathic pulmonary fibrosis, other pulmonary interstitial diseases and other fibrotic, asthmatic or allergic diseases, eosinophilic diseases, chronic obstructive pulmonary disease, and rheumatic and atherosclerosis, as well as pain and neurological disorders such as acute pain, postoperative pain, chronic pain and depression, and bladder disorders.

[0038] Furthermore, the present invention relates to the use of compounds of general formula 1 for the treatment and / or prevention of: (1) Coughs such as chronic idiopathic cough or chronic intractable cough, asthma, COPD, lung cancer, post-viral infection and cough associated with idiopathic pulmonary fibrosis and other pulmonary interstitial diseases. (2) Pulmonary fibrotic diseases such as pneumonia or interstitial pneumonia associated with collagen diseases. Examples of collagen diseases include lupus erythematosus, systemic sclerosis, rheumatoid arthritis, polymyositis and dermatomyositis, idiopathic interstitial pneumonia such as 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 pneumonia due to occupational exposure such as asbestosis, silicosis, miner's lung (coal dust), farmer's lung (hay and mold), pigeon breeder's lung (birds), or interstitial pneumonia due to metal dust or mycobacteria. These include interstitial pneumonias with a clear cause such as interstitial pneumonia due to occupational airborne or other triggers such as pulmonary embolism, or interstitial pneumonia as a result of treatment such as radiation, methotrexate, amiodarone, nitrofurone or chemotherapy, or granulomatous diseases such as granulomatosis with polyangiitis, Churg-Strauss syndrome, sarcoidosis, hypersensitivity pneumonitis, or interstitial pneumonia caused by different causes such as aspiration, inhalation of toxic gases, vapors, bronchitis or pneumonitis or interstitial pneumonia caused by heart failure, x-rays, radiation, chemotherapy, M. boeck or sarcoidosis, granulomatous diseases, cystic fibrosis or pancreatic 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 hepatic bridging fibrosis, liver cirrhosis, nonalcoholic steatohepatitis (NASH), atrial fibrosis, endomyocardial fibrosis, old myocardial infarction, glial scar, atherosclerosis, fibrous arthritis, Dupuytren's contracture, keloid, scleroderma / systemic sclerosis, mediastinal fibrosis, myelofibrosis, Peyronie's disease, nephrogenic systemic fibrosis, retroperitoneal fibrosis, and adhesive arthritis. (4) Inflammatory, autoimmune or allergic diseases and conditions, such as allergic or non-allergic rhinitis or sinusitis, chronic sinusitis or rhinitis, nasal polyps, chronic rhinosinusitis, acute rhinosinusitis, asthma, childhood asthma, allergic bronchitis, alveolitis, hypersensitive airways, allergic conjunctivitis, bronchiectasis, adult respiratory syndrome, bronchial and pulmonary edema, bronchitis or pneumonitis, eosinophilic cellulite (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, etc.; exercise-induced bronchoconstriction; chronic obstructive pulmonary disease. chronic bronchitis, cough, emphysema; systemic anaphylaxis or hypersensitivity reactions, drug allergies (e.g. to penicillins, cephalosporins), eosinophilic polymyalgia syndrome due to ingestion of contaminated tryptophan, insect bite allergies; autoimmune diseases, e.g. rheumatoid arthritis, Graves' disease, Sjogren's syndrome, psoriatic arthritis, multiple sclerosis, systemic lupus erythematosus, gravitational myasthenia, 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 (Rh disease), Goodpasture's syndrome (anti-GBM disease), celiac disease, autoimmune cardiomyopathy, juvenile onset diabetes; glomerulonephritis, autoimmune thyroiditis, Behcet's disease; graft rejection (e.g. at the time of transplantation) including allograft rejection or graft-host disease; inflammatory bowel diseases such as Crohn's disease and ulcerative colitis; spondyloarthropathy; scleroderma; inflammatory skin diseases such as psoriasis (including T-cell psoriasis) and dermatitis, eczema, atopic dermatitis, allergic contact dermatitis, urticaria; vasculitis (e.g. necrotizing, cutaneous and hypersensitivity vasculitis); erythema nodosum; eosinophilic myositis, eosinophilic fasciitis, cancer with infiltration of skin or organs with leukocytes;Ophthalmic diseases such as age-related macular degeneration, diabetic retinopathy and diabetic macular edema, keratitis, eosinophilic keratitis, keratoconjunctivitis, vernal keratoconjunctivitis, eye disorders, anterior eye disorders, blepharitis, blepharoconjunctivitis, bullous disease, cicatricial pemphigoid, conjunctival melanoma, papillary conjunctivitis, dry eye, episcleritis, glaucoma, gliosis, granuloma annulare, Graves' ophthalmopathy, intraocular melanoma, pinguecula, proliferative vitreous retinopathy, pterygium, scleritis, uveitis, acute gout attacks, gout or ocular osteoarthritis; (5) Pain such as chronic idiopathic pain syndrome, neuropathic pain, paresthesia, allodynia, migraine, toothache, and postoperative pain. (6) Depression, anxiety neurosis, diabetic neuropathy, and bladder disorders such as bladder outlet obstruction, overactive bladder, and cystitis; myocardial reperfusion injury or cerebral ischemic injury.

[0039] In a further aspect, the present invention relates to compounds of general formula 1 for use in the treatment and / or prevention of the abovementioned diseases and conditions. In a further aspect the present invention relates to the use of compounds of general formula 1 for the preparation of medicaments for the treatment and / or prevention of the abovementioned 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, which method comprises the administration to a human of an effective amount of a compound of general formula 1.

[0040] Combination Therapy The compounds of the present invention may further be combined with one or more, preferably one, additional therapeutic agent.According to one embodiment, the additional therapeutic agent is selected from the group of therapeutic agents useful in the treatment of diseases or conditions described hereinabove, particularly related to 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 pulmonary interstitial diseases, asthma or allergic diseases, eosinophilic diseases, chronic obstructive pulmonary disease, atopic dermatitis, and autoimmune diseases such as rheumatoid arthritis and atherosclerosis, or therapeutic agents useful in the treatment of ophthalmological diseases, pain and depression. Additional therapeutic agents suitable for such combinations include, in particular, those that enhance the therapeutic effect of one or more active substances for, for example, one of the described indications and / or those that 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, anti-anxiety agents, sedatives, skeletal muscle relaxants or antidepressants.

[0041] Examples of anti-fibrotic agents include phosphodiesterase-IV (PDE4) inhibitors such as nintedanib, pirfenidone, and roflumilast; autotaxin inhibitors such as GPLG-1690 and 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; and c-Jun inhibitors such as CC-90001. N-terminal kinase (JNK) inhibitors; galectin-3 inhibitors such as TD-139; G protein-coupled receptor 84 (GPR84) inhibitors such as GLPG-1205; dual G protein-coupled receptor 84 / G protein-coupled receptor 40 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; prostaglandin synthase inhibitors such as ZL-2102 / SAR-191801; 15-hydroxy-eicosapentaenoic acid acid (15-HEPE, e.g. DS-102); lysyl oxidase-like 2 (LOXL2) inhibitors such as PAT-1251, PXS-5382 / PXS-5338; dual phosphoinositide 3-kinase (PI3K) / mammalian target of rapamycin (mTOR) 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 RNAs such as TRK250 / BNC-1021; or lysophosphatidic acid receptor antagonists such as BMS-986278.

[0042] Antitussives include, for example, purinergic receptor 3 (P2X3) receptor antagonists such as gefapixand, S-600918, BAY-1817080, or BLU-5937; neurokinin 1 (NK-1) receptor antagonists such as orbepitant or aprepitant; nicotinic acetylcholine receptor alpha 7 subunit stimulants such as ATA-101 / bradanicline; codeine, gabapentin, pregablin, or azithromycin. Anti-inflammatory agents include, for example, corticosteroids such as prednisolone or dexamethasone; cyclo-oxygenase-2 (COX2) inhibitors such as celecoxib, rofecoxib, parecoxib, valdecoxib, deracoxib, etoricoxib, or lumiracoxib; prostaglandin E2 antagonists; leukotriene B4 antagonists; leukotriene D4 antagonists such as montelukast; 5-lipoxygenase inhibitors; or other nonsteroidal anti-inflammatory drugs (NSAIDs), such as aspirin, diclofenac, diflunisal, etodolac, ibuprofen, or indomethacin.

[0043] Anti-atopic dermatitis agents include, for example, cyclosporine, methotrexate, phenolate mofetil, azathioprine, phosphodiesterase inhibitors (e.g., apremilast, crisaborole), Janus-associated kinase (JAK) inhibitors (e.g., tofacitinib), IL-4 / IL-13 (e.g., dupilamab), IL-13 (e.g., lebrikizumab, tralokinumab), and neutralizing antibodies against IL-31 (nemolizumab). Examples of analgesics include morphine, oxymorphine, levopane, oxycodone, propoxyphene, nalmefene, fentanyl, hydrocodone, hydromorphone, mellipidine, methadone, nalorphine, naloxone, naltrexone, and buprenorphine. opioid types, such as butorphanol, nalbuphine, pentazocine, etc.; or non-opioid types, such as acetophenamine, etc.

[0044] Examples of antidepressants include tricyclic antidepressants such as amitriptyline, clomipramine, despramine, doxepin, desipramine, imipramine, and nortriptyline; selective serotonin reuptake inhibitor antidepressants (SSRIs) such as fluoxetine, paroxetine, sertraline, citalopram, and escitalopram; maprotiline, lofepramine, mirtazapine, oxaprotiline, fezolamine, tomoxetine, mianserin, buproprion, hydroxybuproprion, and nomifen. norepinephrine reuptake inhibitor antidepressants (SNRIs) such as serotonin, benzodiazepine, benzoxazine, benzoxazine; 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. Anti-anxiety agents include, for example, benzodiazepines such as alprazolam, bromazepam, chlordiazepoxide, clonazepam, clorazepate, diazepam, flurazepam, lorazepam, oxazepam, temazepam, triazolam, or tofizopam; or non-benzodiazepine hypnotics such as eszopiclone, zaleplon, zolpidem, or zopiclone; or carbamates such as meprobamate, carisoprodol, tibamate, or lorbamate; or antihistamines such as hydroxyzine, chlorpheniramine, or diphenhydramine. Examples of analgesics include barbiturate sedatives such as amobarbital, aprobarbital, butabarbital, butabital, mephobarbital, metharbital, methohexital, pentobarbital, secobarbital, talbutal, theamylal, or thiopental; or non-barbiturate sedatives such as glutethimide, meprobamate, methaqualone, or dichloalphenazone. Skeletal muscle relaxants include, for example, baclofen, meprobamate, carisoprodol, cyclobenzaprine, metaxalone, methocarbamol, tizanidine, chlorzoxazone, orphenadrine, and the like.

[0045] Other suitable combination partners are acetylcholinesterase inhibitors, such as donepezil; 5-HT-3 antagonists, such as ondansetron; metabotropic glutamate receptor antagonists; antiarrhythmic drugs, such as mexiletine or phenytoin; or NMDA receptor antagonists. Further suitable combination partners are incontinence drugs, for example anticholinergics such as oxybutynin, tolterodine, darifenacin, fesoterodine, solifenacin or trospium; or bladder muscle relaxants such as mirabegron; or alpha receptor blockers such as tamsulosin, alfuzosin, silodosin, doxazosin or terazosin. The dosage of the above combination patterns is usually from 1 / 5 of the normally recommended minimum dose to 1 / 1 of the normally recommended dose. Thus, in another aspect, the present invention relates to the use of the compounds of the present invention in combination with one or more additional therapeutic agents as described hereinbefore and hereinafter for the treatment of diseases or conditions that may be affected or mediated by TRPA1, in particular the diseases or conditions as described hereinbefore and hereinafter.

[0046] In a further aspect, the present invention relates to a method of treating a disease or condition that can be affected by inhibition of TRPA1 in a patient, comprising the step of 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 yet another 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. In yet another aspect, the present 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 present invention in combination with a therapeutically effective amount of one or more additional therapeutic agents as described hereinbefore and hereinafter.

[0047] The use of the compounds according to the invention in combination with an additional therapeutic agent may be simultaneous or staggered. The compound according to the invention and the one or more additional therapeutic agents may both be present together in one formulation, e.g. a tablet or capsule, or may be present 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 hereinbefore and hereinafter, optionally together with one or more 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 embodiments which illustrate, by way of example, the principles of the invention.

[0048] preparation The compounds according to the invention and intermediates thereof 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 more fully hereinafter, in particular as described in the experimental section.In some cases, the order of carrying out the reaction steps may be changed.Variations of the reaction methods known to those skilled in the art but not described in detail here may also be used. The general processes for preparing compounds according to the invention will be apparent to those skilled in the art upon studying the following schemes. Any functional groups of the starting materials or intermediates may be protected using conventional protecting groups. These protecting groups can be cleaved again at an appropriate 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 herein below, where the substituents of the general formula have the meanings given herein above.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.

[0049] Scheme 1: [ka] In Scheme 1, compounds of formula I can be synthesized via N-alkylation of intermediate (A) with chloromethylene-oxadiazole (B) in the presence of a base such as potassium carbonate.

[0050] Scheme 2: [ka] In Scheme 2, uracil derivative (C) (CAS: 154942-22-0) can be synthesized from methyl urea and 1,3-diethyl 2-(ethoxymethylidene)propanedioate at elevated temperature under solvent-free conditions. Primary amide (A) can be synthesized from ester (C) by stirring with ammonia in a sealed vessel at elevated temperature in a solvent such as water or alcohol.

[0051] Scheme 3: [ka] In Scheme 3, alpha-cyanoketones (E) synthesized from carboxylic esters (D) are enantioselectively reduced to alcohols (F) using an appropriate catalytic system based on transition metal complexes (e.g., Ru or Ir) in combination with chiral ligands (e.g., [(1S,2S)-2-amino-1,2-diphenylethyl](4-toluenesulfonyl)amide) and a hydrogen source such as triethylamine formate complex. These alcohols (F) can be ring-closed to chloromethylene-oxadiazoles (B) by stirring the reaction mixture with chloroacetyl chloride in the presence of a base such as DIPEA. EXAMPLES

[0052] preparation The compounds according to the invention and intermediates thereof can be obtained using synthetic methods known to those skilled in the art and described in the organic synthesis literature, for example, the methods described in “Comrehensive 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 analogously to the preparation methods described more fully hereinafter, in particular those described in the experimental section. In some cases, the order of carrying out the reaction schemes may be changed. Variants of these reactions known to those skilled in the art but not described in detail herein may also be used. The general steps for preparing the compounds according to the invention will be clear to the skilled person on studying the schemes that follow. The starting compounds may be commercially available or may be prepared by methods described in the literature or herein, or may be prepared in a similar or similar manner. Before carrying out the reaction, any corresponding functional groups in the starting compounds may be protected using conventional protecting groups. These protecting groups can be cleaved again at an appropriate 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 refer to a temperature of about 20° C., e.g., 19-24° C.

[0053] Abbreviation: [Table 10-1] [Table 10-2]

[0054] Preparation of intermediates Intermediate I Intermediate I.1 (general route) (3S)-3-(4-chlorophenyl)-3-hydroxypropanenitrile [ka] Under an inert atmosphere, 10.0 g (55.7 mmol) of 4-chlorobenzoylacetonitrile is added to 100 mL of ACN. 142 mg (0.23 mmol) of chloro([(1S,2S)-2-amino-1,2-diphenylethyl](4-toluenesulfonyl)amido)(mesitylene)ruthenium(II) (CAS 174813-81-1) are added, followed by dropwise addition of 8.30 mL (19.8 mmol) of formate triethylamine complex (5:2). After stirring at RT for 3 h, 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)

[0055] The following compounds are prepared using procedures similar to those described above for intermediate I.1 using the appropriate starting materials. As would be expected by one skilled in the art, these analogous examples may include a variety of general reaction conditions. [Table 11]

[0056] Intermediate II Intermediate II.1 (general route) (3S)-3-(4-chlorophenyl)-N,3-dihydroxypropane imidamide [ka] To 9.82 g (54.1 mmol) of (3S)-3-(4-chlorophenyl)-3-hydroxypropanenitrile (intermediate I.1) in 100 mL of MeOH is added 8.00 mL (136 mmol) of hydroxylamine (50% in water) and the mixture is stirred for 1.5 h at 75° C. After cooling to RT, 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)

[0057] The following compounds are prepared using procedures similar to those described above for intermediate II.1 using the appropriate starting materials. As would be expected by one skilled in the art, these analogous examples may include a variety of general reaction conditions. [Table 12]

[0058] Intermediate III Intermediate III.1 (general route) (1S)-2-[5-(chloromethyl)-1,2,4-oxadiazol-3-yl]-1-(4-chlorophenyl)ethan-1-ol [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 kept stirring for 4 h. After cooling to RT, 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 vacuum. 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)

[0059] The following compounds are prepared using procedures similar to those described above for intermediate III.1 using the appropriate starting materials. As would be expected by one skilled in the art, these analogous examples may include a variety of general reaction conditions. [Table 13]

[0060] Intermediate IV Intermediate IV.1 (general route) 3-(6-fluoro-1-benzothiophen-2-yl)-3-oxopropanenitrile [ka] Under an inert atmosphere, 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 is added with 0.36 g (9.00 mmol) of NaH (60% mixture in mineral oil) at RT. The mixture is heated under reflux and stirred for 16 h, cooled to RT, poured into ice / water (30 mL) and treated with 2 M HCl until pH=1 is reached. EtOAc (20 mL) is added and the phases are separated. The aqueous phase is extracted once more with EtOAc (20 mL), the combined organic phases are 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 (D)

[0061] The following compounds are prepared using procedures similar to those described above for intermediate IV.1 using the appropriate starting materials. As would be expected by one skilled in the art, these analogous examples may include a variety of general reaction conditions. [Table 14]

[0062] Intermediate V Ethyl 3-methyl-2,4-dioxo-1,2,3,4-tetrahydropyrimidine-5-carboxylate [ka] 500 mg (6.75 mmol) of methyl urea and 1.36 g (6.75 mmol) of 1,3-diethyl 2-(methoxymethylidene)propanedioate are stirred under neat conditions for 2 h at 120 °C, 17 h at RT, 66 h at 100 °C, 17 h at 150 °C, and 17 h at 120 °C. The mixture is then diluted with EtOAc and refluxed. The mixture is slowly cooled to RT and the precipitated intermediate is removed by filtration. C8H 10 N2O4 (M = 198.2 g / mol) ESI-MS: 199[M+H] + R t (HPLC): 0.24 min (Method A)

[0063] Intermediate VI 3-Methyl-2,4-dioxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide [ka] 10.0 g (50.46 mmol) of ethyl 3-methyl-2,4-dioxo-1,2,3,4-tetrahydropyrimidine-5-carboxylate (CAS: 154942-22-0, intermediate V) in 33% aqueous ammonia (120 mL) is stirred at 100 °C in a sealed vessel for 10 h. The reaction mixture is cooled to RT and concentrated under reduced pressure. The residue is triturated with ACN, filtered, and dried at 50 °C to give intermediate VI. C6H7N3O3 (M = 169.1 g / mol) ESI-MS: 170[M+H] + R t (HPLC): 0.48 min (Method B)

[0064] Preparation of final compounds Example 1 (General Procedure) 1-({3-[(2S)-2-(4-chlorophenyl)-2-hydroxyethyl]-1,2,4-oxadiazol-5-yl}methyl)-3-methyl-2,4-dioxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide [ka] A mixture of 19 mg (0.11 mmol) of intermediate VI, 30 mg (0.11 mmol) of intermediate III.1, and 30 mg (0.22 mmol) of KCO in 1.0 mL of DMF is stirred at RT for 1 h. The reaction mixture is filtered and the filtrate is purified by reverse phase HPLC (ACN / H2O gradient, 0.1% TFA) to give the desired compound. C 17 H 16 ClNO5(M=405.79g / mol) ESI-MS: 406[M+H] + R t (HPLC): 0.44 min (Method A) 1 H NMR(400MHz,DMSO-d6)δppm:2.92-3.07(m,2H),3.23(s,3H),4.96(dd,J=7.9,5.8Hz,1H),5.48 (d,J=1.9Hz,2H),7.31-7.40(m,4H),7.65(d,J=3.3Hz,1H),8.19(d,J=3.3Hz,1H),8.80(s,1H)

[0065] The following compounds are prepared using the appropriate starting materials using procedures similar to the general procedures described above in Example 1. As would be expected by one skilled in the art, these analogous examples may include a variety of general reaction conditions. [Table 15]

[0066] Analytical data for the compounds listed in the table above: [Table 16-1] [Table 16-2]

[0067] Analytical HPLC method

Table 17

Table 18

Table 19

Table 20

[0068]

Table 21

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Claims

1. A compound of formula (I). 【Chemistry 1】 (I) (In the formula, A is selected from the group consisting of phenyl, thiophenyl, benzofuranyl, and benzothiophenyl, and A is unsubstituted or selected from the group consisting of halogen and C 1-4 one or two radicals R consisting of -alkyl 1 has been replaced by .)

2. R 1 is F, Cl, I, and CH 3 2. The compound of formula (I) according to claim 1, selected from:

3. A is the following group: 【Chemistry 2】 and A is unsubstituted or one or two groups R 1 2. A compound of formula (I) according to claim 1, substituted with:

4. A compound comprising: 【Chemistry 3-1】 【change】 【Chemistry 3-2】 【change】 【change】 【change】 【change】 2. The compound of formula (I) according to claim 1, which is a compound selected from the group consisting of:

5. A pharma- ceutically acceptable salt of the compound according to any one of claims 1 to 4.

6. A pharmaceutical composition comprising at least one compound of formula (I) according to any one of claims 1 to 4, or a pharma- ceutically acceptable salt thereof, and one or more pharma- ceutically acceptable excipients.

7. A composition comprising a compound of formula (I) according to any one of claims 1 to 4, or a pharma- ceutically acceptable salt thereof, for use as a medicament.

8. A pharmaceutical composition comprising a compound according to any one of claims 1 to 4, or a pharma- ceutically acceptable salt thereof, for the treatment or prevention of an inflammatory airway disease or a fibrotic disease or cough.

9. A pharmaceutical composition comprising a compound according to any one of claims 1 to 4, or a pharma- ceutically acceptable salt thereof, for the treatment or prevention of idiopathic pulmonary disease (IPF) or cough.

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