Tetraazole derivatives as TRPA1 inhibitors

Abstract

This invention provides certain tetrazolium derivatives that are inhibitors of transient receptor potential ankylosing protein 1 (TRPA1), and are therefore useful for treating diseases that can be treated by inhibiting TRPA1. Pharmaceutical compositions containing these derivatives and methods for preparing said compounds are also provided.

Description

Technical Field

[0001] This invention provides certain tetrazolium derivatives that are inhibitors of transient receptor potential ankylosing protein 1 (TRPA1), and are therefore useful for treating diseases that can be treated by inhibiting TRPA1. Pharmaceutical compositions containing these derivatives and methods for preparing said compounds are also provided. Background Technology

[0002] Transient receptor potential (TRP) channels are a group of voltage-gated ion channels primarily located on the plasma membranes of numerous mammalian cell types. Approximately 30 structurally related TRP channels exist, divided into the following groups: TRPA, TRPC, TRPM, TRPML, TRPN, TRPP, and TRPV. Member A of the transient receptor potential cation channel subfamily, TRPA1 (TRPA1), also known as transient receptor potential ankyrin 1, is the sole member of the TRPA gene subfamily. Structurally, TRPA channels are characterized by multiple N-terminal ankyrin repeat sequences (approximately 14 at the N-terminus of human TRPA1), hence the "A" in the ankyrin name (Montell, 2005).

[0003] TRPA1 is highly expressed in the plasma membranes of sensory neurons in the dorsal root ganglia and nodal ganglia of the skin and lungs, as well as in cells of the small intestine, colon, pancreas, skeletal muscle, heart, brain, bladder, and lymphocytes (https: / / www.proteinatlas.org / ) and human pulmonary fibroblasts.

[0004] TRPA1 is the best known sensor of environmental stimuli, generating somatic sensory modalities such as pain, cold, and itching. TRPA1 is activated by a variety of reactive electrophilic stimuli (e.g., allyl isothiocyanate, reactive oxygen species) and non-reactive compounds (e.g., icilin), involving cough associated with asthma, chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis (IPF), or post-viral cough, as well as chronic idiopathic cough and cough in sensitive patients (Song and Chang, 2015; Grace and Belvisi, 2011). Based on studies demonstrating cough-induced TGF-β elevation, TRPA1 inhibitors could be used to treat IPF, where cough is very common due to its relationship with lung injury (Xie et al., 2009; Froese et al., 2016; Tschumperlin et al., 2003; Yamamoto et al., 2002; Ahamed et al., 2008). TRPA1 antagonists inhibit calcium signaling triggered by cough-like triggers such as oxidative stress from cigarette smoke extract (CSE), inflammatory mediator release, and downregulation of antioxidant gene expression (Lin et al., 2015; Wang et al., 2019). TRPA1 antagonists have been effective in studies related to atopic dermatitis (Oh et al., 2013; Wilson et al., 2013), contact dermatitis (Liu et al., 2013), psoriasis-related pruritus (Wilson et al., 2013), and IL-31-dependent pruritus (Cevikbas et al., 2014). Gain of TRPA1 function in humans has been associated with familial intermittent pain syndrome (Kremeyer et al., 2010). TRPA1 antagonists have been effective in behavioral models of migraine-related abnormal pain (Edelmayer et al., 2012). TRPA1 expression was selectively increased in the trigeminal ganglion innervating damaged teeth compared to its expression in the ganglion innervating healthy teeth (Haas et al., 2011). Several anesthetics are known to be TRPA1 agonists, including isoflurane (Matta et al., 2008), providing a theoretical basis for TRPA1 inhibitors to alleviate postoperative pain. TRPA1 knockout mice and wild-type mice treated with TRPA1 antagonists exhibited anxiolytic and antidepressant-like phenotypes (de Moura et al., 2014). Based on studies showing a mechanistic link between inverse regulation of AMPK and TRPA1, TRPA1 inhibitors are expected to have benefits in the treatment of diabetic neuropathy (Hiyama et al., 2018; Koivisto and Pertovaara, 2013; Wang et al., 2018).Compared to wild-type mice, TRPA1 knockout mice exhibited smaller myocardial infarction size (Conklin et al., 2019). TRPA1 knockout and pharmacological intervention inhibited TNBS-induced colitis in mice (Engel et al., 2011). In a mouse model of cerebral ischemia, TRPA1 knockout and TRPA1 antagonists reduced myelin sheath damage (Hamilton et al., 2016). In a mouse model of gout with monosodium urate, TRPA1 knockout mice showed reduced urate crystals and joint inflammation (Moilanen et al., 2015). In a rat model of acute gout attacks, TRPA1 deficiency in rats improved joint inflammation and hyperalgesia (Trevisan et al., 2014). TRPA1 activation induces an inflammatory response in osteoarthritis chondrocytes (Nummenmaa et al., 2016). TRPA1 inhibition and gene deletion reduced inflammatory mediators in mouse osteoarthritis chondrocytes and mouse cartilage (Nummenmaa et al., 2016). Finally, TRPA1 knockout mice showed improved weight-bearing on osteoarthritis-affected limbs in a MIA-induced knee swelling model (Horvath et al., 2016). TRPA1 was differentially expressed in rat bladder epithelium (Du et al., 2007) and in patients with bladder outlet obstruction (Du et al., 2008). TRPA1 receptor modulation reduced bladder overactivity in a rat spinal cord injury model (Andrade et al., 2011), and intrathecal administration of TRPA1 antagonists alleviated cyclophosphamide-induced cystitis in rats with hypermic micturition reflexes (Chen et al., 2016).

[0005] Therefore, there is a need to provide potent TRPA1 inhibitors.

[0006] Various structural classes of TRPA1 inhibitors are reviewed in S. Skerratt, Progress in Medicinal Chemistry, 2017, Vol. 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.

[0007] WO2017 / 060488 discloses a compound as a TRPA1 antagonist, which has the following general structural formula:

[0008]

[0009] The TRPA1 activity of examples 28 and 29, which contain a tetrazolium ring, was not disclosed.

[0010] L. Schenkel et al., J. Med. Chem. 2016, 59, 2794-2809 disclose quinazolinone-based TRPA1 antagonists, which include compounds with the following general structural formulas:

[0011]

[0012] Compound 31, in which R is disclosed as OH, exhibited IC50 in FLIPR analysis. 50 It exhibits TRPA1 antagonistic activity of 58 nM and has an intrinsic clearance rate of <14 μL / min / kg in human liver microsomes. Detailed Implementation

[0013] This invention discloses a novel tetrazolium derivative that is an inhibitor of transient receptor potential ankylosing protein 1 (TRPA1) and has suitable pharmacological and pharmacokinetic properties, making it suitable as a medicament for treating symptoms and / or diseases that can be treated by inhibiting TRPA1.

[0014] The compounds of the present invention can provide several advantages, such as enhanced potency, high metabolic and / or chemical stability, high selectivity, safety and tolerability, enhanced solubility, enhanced permeability, desired plasma protein binding, enhanced bioavailability, suitable pharmacokinetic morphology and the possibility of forming stable salts.

[0015] The compounds of the present invention

[0016] This invention provides a novel tetrazolium derivative that unexpectedly acts as a potent inhibitor of TRPA1 (Analysis A), and is further characterized by:

[0017] - Modified stability in human liver microsomes (Analysis B)

[0018] - Improved stability in human hepatocytes (Analysis C)

[0019] The compounds of the present invention differ structurally from Examples 28 and 29 in WO2017 / 060488 in that they have a furano[2,3-d]pyridazinyl core and a substituent adjacent to a secondary aliphatic alcohol. Furthermore, the compounds of the present invention differ structurally from Example 31 in L. Schenkel et al., J. Med. Chem. 2016, 59, 2794-2809 in that they have a tetrazolium ring. These structural differences unexpectedly lead to the following advantageous combination: (i) inhibition of TRPA1, (ii) stability in human liver microsomes, and (iii) stability in human hepatocytes.

[0020] Therefore, the compounds of the present invention are superior to the compounds disclosed in the prior art in terms of the combination of the following parameters:

[0021] -Efficacy as a TRPA1 inhibitor

[0022] -Stability in human liver microsomes

[0023] -Stability in human liver cells.

[0024] Stability in human liver microsomes refers to the sensitivity of a compound to biotransformation in the case of selecting and / or designing drugs with favorable pharmacokinetic properties as the first screening step. The liver is the primary site of metabolism for many drugs. 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 an appropriate half-life, thereby allowing for less frequent and less frequent dosing by patients. Therefore, enhanced stability in human liver microsomes is an advantageous feature for compounds intended for pharmaceutical use. Thus, in addition to the ability to inhibit TRPA1, the compounds of the present invention are expected to have favorable in vivo clearance in humans and therefore the desired duration of action.

[0025] Stability in human hepatocytes refers to a compound's sensitivity to biotransformation in the context of selecting and / or designing drugs with favorable pharmacokinetic properties. The liver is the primary site of metabolism for many drugs. Human hepatocytes contain cytochrome P450 (CYP) and other drug-metabolizing enzymes, and therefore represent a model system for studying drug metabolism in vitro. (Importantly, compared to liver microsomal analysis, hepatocyte analysis also encompasses phase II biotransformation and processes mediated by liver-specific transporters, and therefore represents a more complete system for drug metabolism studies.) Enhanced stability in human hepatocytes is associated with several advantages, including increased bioavailability and an appropriate half-life, thereby enabling less frequent and fewer doses to be administered to patients. Therefore, enhanced stability in human hepatocytes is an advantageous feature for compounds intended for use as drugs.

[0026] This invention provides novel compounds according to formula (I).

[0027]

[0028] in

[0029] A is selected from the group consisting of: phenyl, thienyl, benzothienyl, and benzofuranyl, which are unsubstituted or derived from group R. 3 One or two members are replaced, and the group consists of the following: -CN, halogen, C 1-4 Alkyl, OC 1-4 Alkyl, C 1-4 Fluoroalkyl, OC 1-4 fluoroalkyl, C 3-4 cycloalkyl, OC 3-4 cycloalkyl, C 3-4 Cyclofluoroalkyl and OC 3-4 Cyclofluoroalkyl;

[0030] R 1 It can be H, H3C or H2N(O)C;

[0031] and

[0032] R 2 It is H3C or H2N(O)C;

[0033] Its limitations are:

[0034] When R 1 When it is H2N(O)C, R 2 For H3C;

[0035] When R 2 When it is H2N(O)C, R 1 It is H or H3C.

[0036] Another embodiment of the present invention relates to a compound of formula (I), wherein

[0037] A. Choose from the following groups:

[0038]

[0039] and

[0040]

[0041] It was not replaced or passed through the group R 3 One or two members are replaced;

[0042] And the substituent R 1 and R 2 As defined in any of the foregoing implementation schemes.

[0043] Another embodiment of the invention relates to a compound of formula (I), wherein R 3 Choose from the following groups: Br, Cl, F, and H3C, with substituents A and R. 1 and R 2 As defined in any of the foregoing implementation schemes.

[0044] Another embodiment of the present invention relates to a compound of formula (I), wherein

[0045] A. Choose from the following groups:

[0046]

[0047]

[0048] and

[0049]

[0050] And the substituent R 1 and R 2 As defined in any of the foregoing implementation schemes.

[0051] Another embodiment of the present invention relates to a compound of formula (I), wherein

[0052] R 1 H3C and R 2 It is H2N(O)C;

[0053] And substituents A and R 3 As defined in any of the foregoing implementation schemes.

[0054] Another embodiment of the present invention relates to a compound of formula (I), wherein

[0055] R 1 It is H2N(O)C and R 2 For H3C;

[0056] And substituents A and R 3 As defined in any of the foregoing implementation schemes.

[0057] Compounds of formula (I) preferably selected from the group consisting of the following:

[0058]

[0059] and

[0060]

[0061] And the substituent A is as defined in any of the foregoing embodiments.

[0062] Compounds according to formula (I) that are selected from the group consisting of the following groups are particularly preferred:

[0063]

[0064]

[0065]

[0066] and

[0067]

[0068] Terms and definitions used

[0069] Terms not specifically defined herein shall have the meanings that those skilled in the art would interpret based on the disclosure and context. However, unless otherwise stated, the following terms shall have their designated meanings as used herein and shall follow the conventions outlined below.

[0070] In the groups, radicals, or portions defined below, the number of carbon atoms is usually specified before the group, e.g., C 1-6 Alkyl refers to an alkyl group / alkyl radical having 1 to 6 carbon atoms. Generally, in groups such as HO, H2N, (O)S, (O)2S, NC (cyano), HOOC, F3C, or similar groups, those skilled in the art can see the group's free valence from its own to the group's junction point in the molecule. For composite groups containing two or more daughter groups, the last named daughter group is the group's junction point, for example, the substituent "aryl-C". 1-3 "alkyl" means related to C 1-3alkyl-bonded aryl, the C 1-3 Alkyl groups are attached to the core or to the substituent.

[0071] If the compounds of this invention are described by chemical name and chemical formula, in the event of any discrepancy, the chemical formula shall prevail. An asterisk may be used in the sub-chemical formula to indicate bonds attached to the core molecule as defined.

[0072] The numbering of each atom in a substituent begins with the atom closest to the core or the atom closest to the group to which the substituent is attached.

[0073] For example, the term "3-carboxypropyl-group" indicates the following substituents:

[0074]

[0075] The carboxyl group is attached to the third carbon atom of the propyl group. The terms "1-methylpropyl-", "2,2-dimethylpropyl-", or "cyclopropylmethyl-" represent the following groups:

[0076]

[0077] An asterisk can be used in a sub-chemical formula to indicate a bond attached to a core molecule as defined.

[0078] The term "C" alone or in combination with another group 1-n "Alkyl" refers to a non-cyclic, saturated, branched, or straight-chain hydrocarbon group having 1 to n carbon atoms, where n is an integer selected from 2, 3, 4, or 5. For example, the term C... 1-5 Alkyl groups include 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-CH(CH3)-, 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)- and H3C-CH2-CH(CH2CH3)-.

[0079] The term "fluorine" added to "alkyl," "alkylene," or "cycloalkyl" (saturated or unsaturated) means such alkyl or cycloalkyl groups in which one or more hydrogen atoms are replaced by fluorine atoms. Examples include, but are not limited to, H2FC-, HF2C-, and F3C-.

[0080] The term phenyl refers to a group in the following ring:

[0081]

[0082] The term thiophene group refers to a group that forms a ring with the following ring:

[0083]

[0084] The term benzothiophene group refers to a group with the following ring:

[0085]

[0086] The term benzofuranyl refers to a group with the following ring:

[0087]

[0088] The term furano[2,3-d]pyridazinyl refers to a group on the following ring:

[0089]

[0090] The term tetrazolium refers to a group with the following ring:

[0091]

[0092] As used herein, the term “substituted” means that any one or more hydrogen atoms on a specified atom are selectively replaced from a specified group, provided that the substitution does not exceed the normal valence state of the specified atom and that the substitution produces a stable compound.

[0093] Unless otherwise specified, throughout the specification and the appended claims, the given chemical formula or name shall encompass its tautomers and all stereoisomers, optical isomers and geometric isomers (e.g., mirror-image isomers, non-mirror-image isomers, E / Z isomers, etc.) and racemic mixtures, as well as mixtures of individual mirror-image isomers in different proportions, mixtures of non-mirror-image isomers, or mixtures containing any of the aforementioned forms of such isomers and mirror-image isomers, and salts, including pharmaceutically acceptable salts thereof, and their solvates, such as hydrates, including solvates of the free compound or solvates of salts of the compound.

[0094] Generally, substantially pure stereoisomers can be obtained according to synthetic principles known to those skilled in the art, for example by separating the corresponding mixtures, by using stereochemically pure starting materials, and / or by stereoselective synthesis. Optically active forms are known in the art, such as by resolving racemic forms, or by synthesis, for example, starting from optically active starting materials, and / or by using chiral reagents.

[0095] The mirror-isomer pure compounds or intermediates of the present invention can be prepared via asymmetric synthesis, for example by preparing and subsequently isolating suitable non-mirror-isomer compounds or intermediates, which can be isolated 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.

[0096] Furthermore, those skilled in the art know how to prepare mirror-isomeric pure compounds from corresponding racemic mixtures, such as by chromatographic separation of the corresponding racemic mixture on a chiral stationary phase; or by resolving the racemic mixture using a suitable eluent, for example by forming a non-mirror-isomeric salt with the racemic compound and an optically active acid or base, followed by resolving the salt and releasing the desired compound from the salt; or by derivatizing the corresponding racemic compound with an optically active chiral auxiliary agent, followed by separating the non-mirror-isomer and removing the chiral auxiliary group; or by kinetic analysis of the racemic mixture (e.g., by enzymatic analysis); by mirror-selective crystallization from aggregates of mirror-isomeric crystals under suitable conditions; or by (stepwise) crystallization from a suitable solvent in the presence of an optically active chiral auxiliary agent.

[0097] The phrase “pharmaceutically acceptable” is used in this article to refer to compounds, materials, compositions, and / or dosage forms that are suitable for use within the limits of reasonable medical judgment, without excessive toxicity, irritation, allergic reactions, or other problems or complications, and in proportion to a reasonable benefit / risk ratio.

[0098] As used herein, "pharmaceutically acceptable salt" refers to a derivative of the disclosed compound in which the parent compound forms a salt or complex with an acid or base. Examples of acids that form pharmaceutically acceptable salts with a parent compound containing a basic moiety include inorganic or organic acids such as benzenesulfonic acid, benzoic acid, citric acid, ethanesulfonic acid, fumaric acid, gentian acid, hydrobromic acid, hydrochloric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, 4-methylbenzenesulfonic acid, phosphoric acid, salicylic acid, succinic acid, sulfuric acid, and tartaric acid.

[0099] Examples of cations and bases that form pharmaceutically acceptable salts with parent compounds containing acidic moieties include Na. + K + Ca 2+ Mg 2+ NH4 + L-arginine, 2,2'-imine diethanol, L-lysine, N-methyl-D-glucosamine, or tris(hydroxymethyl)-aminomethane. The pharmaceutically acceptable salts of the present invention can be synthesized from parent compounds containing a basic or acidic moiety using conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or free base form of these compounds with a sufficient amount of a suitable base or acid in water or in an organic diluent such as diethyl ether, ethyl acetate, ethanol, isopropanol, or acetonitrile, or mixtures thereof.

[0100] Salts of acids other than those mentioned above (e.g., trifluoroacetates) that can be used, for example, to purify or separate the compounds of the present invention are also part of the present invention.

[0101] Bioanalysis

[0102] Assessment of TRPA1 activity

[0103] Analysis A: TRPA1 analysis

[0104] The activity of the compounds of the present invention can be confirmed using the following in vitro TRPA1 cell assays:

[0105] method:

[0106] Human HEK293 cell line (Perkin Elmer, product number AX-004-PCL) overexpressing the human TRPA1 ion channel was used as the assay system for compound efficacy and potency. Compound activity was determined by measuring the effect of the compound on intracellular calcium concentration induced by the agonistic effect of allylisothiocyanate (AITC) in a FLIPRtetra system (Molecular Devices).

[0107] Cell culture:

[0108] Cells were obtained in cryo-vitroscopy as frozen cells and stored at -150°C for later use.

[0109] Cells were grown in MEM / EBSS medium containing 10% FCS and 0.4 mg / mL geneticin. Importantly, the cell density should not exceed 90% confluence. For subculture, cells were isolated from the flasks using Versene. The day before analysis, cells were isolated, washed twice with medium (MEM / EBSS medium containing 10% FCS), and 20,000 cells were seeded at 20 μL per well into 384-well poly-D-lysine biocoated discs (black, clear bottom, catalog number 356697) from Corning. The discs were incubated at 37°C / 5% CO2 for 24 hours before analysis.

[0110] Compound preparation

[0111] The test compound was dissolved in 100% DMSO at a concentration of 10 mM, and then diluted to a concentration of 5 mM in DMSO in the first step, followed by successive dilution steps in 100% DMSO. The dilution factor and the number of dilution steps can be varied as needed. Typically, eight different concentrations are prepared at a 1:5 dilution ratio, and further intermediate dilutions (1:20) are performed using HBSS / HEPES buffers (1×HEPES from Gibco, catalog number 14065; 20 mM HEPES from SIGMA, catalog number 83264; 0.1% BSA at pH 7.4 from Invitrogen, catalog number 11926).

[0112] FLIPR analysis:

[0113] On the analysis day, cells were washed three times with analysis buffer, leaving 20 μL of buffer residue in the wells after washing. 10 μL of loading buffer from the Ca6 kit (catalog number R8191 MolecularDevices) in HBSS / HEPES was added to the cells, and the trays were incubated at 37°C / 5% CO2 for 120 min with the lid on. 10 μL of the compound or control from the intermediate dilution tray in HBSS / HEPES buffer / 5% DMSO was carefully added to the wells. The luminescence (indicating calcium influx or release) was read on a FLIPRtetra device for 10 min to monitor the compound-induced effect (e.g., agonist effect). Finally, 10 μL of 50 μM agonist AITC dissolved in HBSS / HEPES buffer / 0.05% DMSO (final concentration 10 μM) was added to the wells, followed by another 10 min reading on a FLIPRtetra device. The IC50 / inhibition percentage was calculated using the area under the signal curve (AUC) after AITC addition.

[0114] Data evaluation and calculation:

[0115] Each analytical microtitration pan contains a well with a mediator (1% DMSO) control instead of the compound as a control for AITC-induced luminescence (100% CTL; high control), and a well with a mediator control but without AITC as a control for non-specific changes in luminescence (0% CTL; low control).

[0116] Data analysis was performed by calculating the area under the signal curve for each individual well. Based on this value, the percentage value (AUC(sample) - AUC(low)) × 100 / (AUC(high) - AUC(low)) used to measure the concentration of each substance was calculated using MegaLab software (in-house developed). The IC50 value was then calculated from the control value % using MegaLab software. Calculation: [y = (ad) / (1 + (x / c)^b) + d], a = low value, d = high value; x = concentration M; c = IC50 M; b = peak; y = control %

[0117] Table 1: Biological data of the compounds of the present invention obtained from analysis A

[0118]

[0119]

[0120] Table 2: Biological information of the background art compounds (Examples 28 and 29 in WO2017 / 060488) obtained in Analysis A.

[0121]

[0122] Table 3: Biological data of the background technology compound (Example 31 in L. Schenkel et al., J. Med. Chem. 2016, 59, 2794-2809) obtained in Analysis A.

[0123]

[0124] Assessment of microsomal clearance rate

[0125] Analysis B: Microsomal clearance rate:

[0126] The metabolic degradation of the test compound was analyzed using collected liver microsomes at 37°C. At each time point, a final incubation volume of 100 μl contained TRIS buffer pH 7.6 (0.1 M), magnesium chloride (5 mM), microsomal protein (1 mg / ml), and the test compound at a final concentration of 1 μM at room temperature.

[0127] After a short pre-incubation at 37°C, the reaction was initiated by adding β-nicotinamide adenine dinucleotide phosphate (NADPH, 1 mM) in its reduced form and terminated by transferring aliquots to a solvent at different time points (0, 5, 15, 30, and 60 minutes). NADPH-independent degradation in the NADPH-free culture was monitored, and the reaction was terminated at the final time point. The percentage of residual test compounds after NADPH-independent incubation is reflected by parameter c (control) (metabolic stability). The quenched culture was precipitated by centrifugation (10000 g, 5 minutes).

[0128] The amount of the parent compound in aliquots of the supernatant was analyzed by LC-MS / MS. The half-life (t1 / 2 in vitro) was determined by the slope of a semi-logarithmic plot of the concentration-time curve.

[0129] The intrinsic clearance rate (CL_INTRINSIC) is calculated by taking into account the amount of protein in the culture:

[0130] CL_INTRINSIC[μl / min / mg protein]=(Ln 2 / (half-life[min]×protein content[mg / ml]))×1000

[0131] CL_INTRINSIC_INVIVO[ml / min / kg]=(CL_INTRINSIC[μL / min / mg protein]×MPPGL[mg protein / g liver]×liver factor[g / kg body weight]) / 1000

[0132] Qh[%]=CL[ml / min / kg] / liver blood flow[ml / min / kg])

[0133] Human hepatocyte count: 120 × 10⁶ cells / g liver

[0134] Human liver factor: 25.7g / kg body weight

[0135] Human blood flow: 21 ml / (min×kg)

[0136] Table 4: Biological data of the compounds of the present invention obtained from analysis B

[0137]

[0138]

[0139] Table 5: Biological information of the background technology compounds (Examples 28 and 29 in WO2017 / 060488) obtained in analysis B.

[0140]

[0141] Table 6: Biodata of the background technology compound obtained in analysis B (Example 31 in L. Schenkel et al., J. Med. Chem. 2016, 59, 2794-2809).

[0142]

[0143] Assessment of hepatocyte clearance rate

[0144] Analysis C: Hepatocyte clearance rate

[0145] The metabolic degradation of the test compounds was analyzed in hepatocyte suspension. Hepatocytes (cryopreserved) were cultured in Dulbecco modified Eagle medium supplemented with 3.5 μg glucagon / 500 mL, 2.5 mg insulin / 500 mL and 3.75 mg / 500 mL DHEA containing 5% species serum.

[0146] After pre-incubating for 30 minutes in an incubator (37°C, 10% CO2), 5 μl of the test compound solution (80 μM; diluted 1:25 with culture medium from 2 mM in DMSO stock solution) was added to 395 μl of hepatocyte suspension (depending on species, cell density ranges from 0.25 to 5 million cells / mL, typically 1 million cells / mL; final concentration of test compound is 1 μM, final concentration of DMSO is 0.05%).

[0147] Cells were cultured for six hours (incubator, rotary shaker) and samples (25 μl) were collected at 0, 0.5, 1, 2, 4, and 6 hours. The samples were transferred to acetonitrile and centrifuged (5 min) to precipitate the precipitate. The supernatant was transferred to a new 96-well plate, evaporated under nitrogen, and resuspended.

[0148] The reduction of the parent compound was analyzed by HPLC-MS / MS.

[0149] CLint is calculated as follows: CL_INTRINSIC = dose / AUC = (C0 / CD) / (AUD + clast / k) × 1000 / 60. C0: initial concentration in the culture [μM], CD: cell density of live cells [10e6 cells / mL], AUD: area under the data [μM × h], clast: concentration of the last data point [μM], k: slope of the regression line of the parent [h-1].

[0150] The calculated in vitro intrinsic liver clearance can be scaled up to obtain the in vivo intrinsic liver clearance, and using this liver clearance, the in vivo hepatic blood clearance (CL) can be predicted by using a liver model (a well-stirred model).

[0151] CL_INTRINSIC_INVIVO[ml / min / kg] = (CL_INTRINSIC[μL / min / 10e6 cells] × hepatocyte count[10e6 cells / g liver] × liver factor[g / kg body weight]) / 1000

[0152] 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])

[0153] Qh[%]=CL[ml / min / kg] / liver blood flow[ml / min / kg])

[0154] Human hepatocyte count: 120 × 10⁶ cells / g liver

[0155] Human liver factor: 25.7g / kg body weight

[0156] Human blood flow: 21 ml / (min×kg)

[0157] Table 7: Biological data of the compounds of the present invention obtained from analysis C

[0158] Example Human hepatocytes [%Qh] 1 4 2 23 3 5 4 30 5 23 6 24 7 15 8 14 9 16 10 <4 11 8 12 16 13 22 14 42 15 21

[0159] Table 8: Biological information of the background art compounds (Examples 28 and 29 in WO2017 / 060488) obtained in analysis C.

[0160]

[0161] Table 9: Biodata of the background technology compound obtained in analysis C (Example 31 in L. Schenkel et al., J. Med. Chem. 2016, 59, 2794-2809).

[0162]

[0163] Assessing penetration

[0164] Caco-2 cells (1-2×10) 5 Cells / 1cm 2 The inoculum (area) was inoculated onto filter inserts (Costartranswell polycarbonate or PET filters, 0.4 μm pore size) and cultured (DMEM) for 10 to 25 days.

[0165] Dissolve the compound in a suitable solvent (e.g., DMSO, 1–20 mM stock solution). Dilute the stock solution 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 Na2HPO4×7H2O, 0.41 mM NaH2PO4×H2O, 15 mM HEPES, 20 mM glucose, 0.25% BSA, pH 7.2) to prepare transport solutions (0.1–300 μM compound, final DMSO <= 0.5%). Apply the transport solution (TL) to the top or bottom outside the donor side to measure AB or BA permeability, respectively (repeated filtration 3 times). Collect samples from the donor side at the beginning and end of the experiment, and also from the receiver side at various time intervals within up to 2 hours, and determine the concentration by HPLC-MS / MS or scintillation counting. Replace the volume of the receiver after sampling with fresh receiver solution.

[0166] Assess plasma protein binding

[0167] This balanced dialysis (ED) technique is used to determine the approximate in vitro binding of test compounds to plasma proteins. A Dianorm Teflon dialysis cell (0.2 μm) is used. Each cell consists of donor and acceptor chambers separated by an ultrathin semi-permeable membrane with a 5 kDa molecular weight cutoff. A stock solution of each test compound at 1 mM is prepared in DMSO and diluted to a final concentration of 1.0 μM. Subsequent dialysis solutions are prepared from human or rat plasma (containing NaEDTA) collected from male and female donors. Aliquots of 200 μL of dialysis buffer (100 mM potassium phosphate, pH 7.4) are aliquoted into the buffer chamber. Aliquots of 200 μL of the test compound dialysis solution are aliquoted into the plasma chamber. The mixture is incubated at 37°C under rotation for 2 hours.

[0168] At the end of the dialysis period, the dialysate was transferred to a reaction tube. The tube used for the buffer portion contained 0.2 mL ACN / water (80 / 20). A 25 μL aliquot of plasma dialysate was 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 was performed by adding 200 μL ACN. A 50 μL aliquot of buffer dialysate was transferred to a deep-well plate and mixed with 25 μL blank plasma, 25 μL internal standard solution, and 200 μL ACN. The samples were measured on an HPLC-MS / MS system and evaluated using Analyst software. The binding percentage was calculated using the following formula: Binding% = (Plasma concentration - Buffer concentration / Plasma concentration) × 100.

[0169] Assess solubility

[0170] In a well tray (format depends on the robot), a saturated solution is prepared by adding an appropriate volume of the selected aqueous culture medium (typically in the range of 0.25–1.5 ml) to each well containing a known amount of solid active pharmaceutical ingredient (typically in the range of 0.5–5.0 mg). The wells are shaken or stirred for a predetermined period of 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). Filter absorption is avoided by discarding the first few drops of filtrate. The amount of dissolved active pharmaceutical ingredient is determined by UV spectroscopy. Additionally, the pH of the saturated aqueous solution is measured using a glass electrode pH meter.

[0171] Assess pharmacokinetic characteristics

[0172] The test compound was administered intravenously or orally to the respective test species. Blood samples were collected at several time points after administration of the test compound, and then anticoagulated and centrifuged.

[0173] The concentrations of the analytes, i.e., the administered compounds and / or metabolites, in the plasma samples were quantified. PK parameters were calculated using a non-compartmental model method. AUC and Cmax were normalized to a 1 μmol / kg dose.

[0174] Assessment of metabolism in human hepatocytes in vitro

[0175] Primary human hepatocytes in suspension form were used to investigate the metabolic pathways of the test compounds. After recovery from cryopreservation, human hepatocytes were cultured in Dulbecco modified Eagle medium containing 5% human serum supplemented with 3.5 μg glucagon / 500 ml, 2.5 mg insulin / 500 ml, and 3.75 mg / 500 ml DHEA.

[0176] After pre-culturing in a cell culture incubator (37°C, 10% CO2) for 30 minutes, the test compound solution was incorporated into the hepatocyte suspension to obtain 1.0 × 10⁻⁶ cells. 6 Up to 4.0×10 6 The final cell density was determined by the number of cells per milliliter (depending on the metabolic conversion rate of the compound observed using primary human hepatocytes), the final concentration of the test compound was 10 μM, and the final concentration of DMSO was 0.05%.

[0177] Cells were incubated in a cell culture incubator with a horizontal shaker for six hours, and samples were removed from the culture after 0, 0.5, 1, 2, 4, or 6 hours, depending on the metabolic conversion rate. Samples were quenched with acetonitrile and precipitated by centrifugation. The supernatant was transferred to a 96-well deep-well plate, evaporated under nitrogen, and resuspended. Subsequent bioanalysis by liquid chromatography-high resolution mass spectrometry was performed to identify presumed metabolites.

[0178] Based on Fourier-Transform-MS n Data are provisionally structured. Metabolites are reported as a percentage of the parent product in human hepatocyte cultures, with a critical limit of ≥4%.

[0179] Treatment

[0180] This invention relates to compounds of general formula 1, which can be used for the prevention and / or treatment of diseases and / or symptoms associated with or regulated by TRPA1 activity, including but not limited to the treatment and / or prevention of fibrotic diseases, inflammatory and immunomodulatory conditions, respiratory or gastrointestinal diseases or discomforts, ophthalmic diseases, inflammatory diseases of the joints and inflammatory diseases of the nasopharynx, eyes and skin, as well as pain and neurological disorders. The conditions, diseases and discomforts include cough, idiopathic pulmonary fibrosis, other interstitial lung diseases and other fibrotic diseases, asthma or allergic diseases, eosinophilic diseases, chronic obstructive pulmonary disease, and inflammatory and immunomodulatory conditions such as rheumatoid arthritis and atherosclerosis, as well as pain and neurological disorders such as acute pain, surgical pain, chronic pain, depression and bladder disorders.

[0181] Compounds of general formula 1 can be used for prevention and / or treatment:

[0182] (1) Cough, such as chronic idiopathic cough or chronic refractory cough, cough associated with asthma, COPD, lung cancer, post-viral infection and idiopathic pulmonary fibrosis and other interstitial lung diseases.

[0183] (2) Pulmonary fibrotic diseases, such as pneumonia or interstitial pneumonia associated with collagen degeneration, such as lupus erythematosus, systemic scleroderma, 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, cryptogenic organizing pneumonia, acute interstitial pneumonia, and lymphocytic interstitial pneumonia; lymphangioleiomyomatosis; pulmonary alveolar proteinosis; Langerhans cell lineage. Histiocytosis of the pleural parenchyma; pleural fibrosis; interstitial lung diseases of known etiology, such as interstitial pneumonia caused by occupational exposure, such as asbestosis, silicosis, miner's lung (coal dust), farmer's lung (hay and mold), pigeon feeder's lung (birds), or other occupational airborne triggers such as metal dust or mycobacteria, or interstitial pneumonia caused by treatments such as radiation, methotrexate, amiodarone, nitrofurantoin, or chemotherapeutic agents; or granulomatous diseases, such as granulomatous polyangiitis, Churg-Strauss syndrome. Interstitial pneumonia (ISP) caused by various sources (e.g., inhalation of toxic gases, vapors, bronchitis, or pneumonia); or caused by heart failure, X-rays, radiation, chemotherapy, M. boeck syndrome, sarcomatoid disease, granulomatous disease, cystic fibrosis, myxoviscosity, or α-1 antitrypsin deficiency.

[0184] (3) Other fibrotic diseases, such as hepatic bridging fibrosis, cirrhosis, non-alcoholic steatosis (NASH), atrial fibrosis, endocardial myocardial fibrosis, old myocardial infarction, glial scars, arteriosclerosis, joint fibrosis, Dupuytren's contracture, keloids, scleroderma / systemic sclerosis, mediastinal fibrosis, myelofibrosis, Peyronie's disease, renal systemic fibrosis, retroperitoneal fibrosis, and adhesive capsulitis.

[0185] (4) Inflammatory, autoimmune, or allergic diseases and symptoms, 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, airway hyperresponsiveness, allergic conjunctivitis, bronchiectasis, adult respiratory distress syndrome, bronchial and pulmonary edema, bronchitis or pneumonia, eosinophilic cellulitis (e.g., Well's syndrome), eosinophilic pneumonia (e.g., Loeffler's syndrome, chronic eosinophilic pneumonia), eosinophilic fasciitis (e.g., Shulman's syndrome). Hypersensitivity syndrome (HSS), delayed-type hypersensitivity, non-allergic asthma; exercise-induced bronchoconstriction; chronic obstructive pulmonary disease (COPD), acute bronchitis, chronic bronchitis, cough, emphysema; systemic severe allergic reactions or anaphylactic reactions, drug allergies (e.g., allergies to penicillin, cephalosporins), eosinophilia-myalgia syndrome caused by ingestion of contaminated tryptophan, insect sting allergies; autoimmune diseases such as rheumatoid arthritis, Graves' disease, Sjogren's syndrome, psoriatic arthritis, multiple sclerosis, systemic lupus erythematosus, myasthenia gravis, immune thrombocytopenic purpura (ITP in adults, neonatal ITP, pediatric ITP), immune hemolytic anemia (autoimmune and drug-induced), Evans syndrome (immune thrombocytopenic purpura of erythrocytes and corpora cavernosa), neonatal Rh disease, Goodpasture's syndrome. Syndrome (anti-GBM disease), celiac disease, autoimmune cardiomyopathy, juvenile-onset diabetes; glomerulonephritis, autoimmune thyroiditis, Behcet's disease; graft rejection (e.g. in transplantation), including allogeneic graft rejection or graft-versus-host disease; inflammatory bowel diseases, such as Crohn's disease and ulcerative colitis; spondyloarthritis; scleroderma; psoriasis (including T-cell mediated psoriasis) and inflammatory skin diseases, such as dermatitis, eczema, atopic dermatitis, allergic contact dermatitis, rubella; vasculitis (e.g., necrotizing, cutaneous, and allergic 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 keratoconjunctivitis, scarring, anterior segment scarring, blepharitis, palpebral conjunctivitis, bullous diseases, cicatricial pemphigoid, conjunctival melanoma, papillary conjunctivitis, dry eye syndrome, episcleritis, glaucoma, glioma, granuloma annulare, Graves' ophthalmopathy, intraocular melanoma, pinguecula, proliferative vitreoretinopathy, pterygium, scleritis, uveitis, acute gout attacks, gout, or osteoarthritis.

[0186] (5) Pain, such as chronic idiopathic pain syndrome, neuropathic pain, hypoesthesia, tenderness to touch, migraine, toothache and postoperative pain.

[0187] (6) Depression, anxiety, diabetic neuropathy and bladder disorders, such as bladder outlet obstruction, overactive bladder, cystitis; myocardial reperfusion injury or cerebral ischemia injury.

[0188] Therefore, the present invention relates to a compound of general formula 1, which is used as a drug.

[0189] Furthermore, the present invention relates to the use of compounds of general formula 1 for the treatment and / or prevention of diseases and / or symptoms related to or regulated by TRPA1 activity.

[0190] Furthermore, this invention relates to the use of compounds of formula 1 for the treatment and / or prevention of fibrotic diseases, inflammatory and immunomodulatory conditions, respiratory or gastrointestinal diseases or discomforts, ophthalmic diseases, inflammatory diseases of the joints and inflammatory diseases of the nasopharynx, eyes and skin, pain and neurological disorders. The conditions, diseases and discomforts include cough, idiopathic pulmonary fibrosis, other interstitial lung diseases and other fibrotic diseases, asthma or allergic diseases, eosinophilic diseases, chronic obstructive pulmonary disease, and inflammatory and immunomodulatory conditions such as rheumatoid arthritis and atherosclerosis, as well as pain and neurological disorders such as acute pain, surgical pain, chronic pain, depression and bladder disorders.

[0191] Furthermore, this invention relates to the use of compounds of general formula 1 for treatment and / or prevention:

[0192] (1) Cough, such as chronic idiopathic cough or chronic refractory cough, cough associated with asthma, COPD, lung cancer, post-viral infection and idiopathic pulmonary fibrosis and other interstitial lung diseases.

[0193] (2) Pulmonary fibrotic diseases, such as pneumonia or interstitial pneumonia associated with collagen degeneration, such as lupus erythematosus, systemic scleroderma, 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, cryptogenic organizing pneumonia, acute interstitial pneumonia, and lymphocytic interstitial pneumonia; lymphangioleiomyomatosis; pulmonary alveolar proteinosis; Langerhans cell histiocytosis; pleural parenchymal elastic fiber hyperplasia; interstitial lung diseases of known etiology, such as interstitial pneumonia caused by occupational exposure, such as asbestosis, silicosis, miner's lung (coal dust), and farmer's lung (hay and mold). Interstitial pneumonia caused by airborne triggers in pigeon breeders (birds) or other occupational sources such as metal dust or mycobacteria, or by treatments such as radiation, methotrexate, amiodarone, nitrofurantoin, or chemotherapeutic agents; or granulomatous diseases such as granulomatous polyangiitis, Churg-Strauss syndrome, sarcoidosis, hypersensitivity pneumonia, or interstitial pneumonia caused by various sources (e.g., inhalation, aspiration of toxic gases, vapors, bronchitis, or pneumonia); or by heart failure, X-rays, radiation, chemotherapy, M. boeck or sarcoidosis, granulomatous disease, cystic fibrosis or myxoviscosity, or α-1 antitrypsin deficiency.

[0194] (3) Other fibrotic diseases, such as hepatic bridging fibrosis, cirrhosis, non-alcoholic steatosis (NASH), atrial fibrosis, endocardial myocardial fibrosis, old myocardial infarction, glial scars, arteriosclerosis, joint fibrosis, DePietrond's contracture, keloids, scleroderma / systemic sclerosis, mediastinal fibrosis, myelofibrosis, Peloni's disease, renal systemic fibrosis, retroperitoneal fibrosis, and adhesive capsulitis.

[0195] (4) Inflammatory, autoimmune, or allergic diseases and symptoms, 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, airway hyperresponsiveness, allergic conjunctivitis, bronchiectasis, adult respiratory distress syndrome, bronchial and pulmonary edema, bronchitis or pneumonia, eosinophilic cellulitis (e.g., Wilms' syndrome), eosinophilic pneumonia (e.g., Lovell's syndrome, chronic eosinophilic pneumonia), eosinophilic fasciitis (e.g., Shulman syndrome), delayed-type hypersensitivity, non-allergic asthma; exercise-induced bronchoconstriction. Shortening; Chronic obstructive pulmonary disease (COPD), acute bronchitis, chronic bronchitis, cough, emphysema; systemic severe allergies or anaphylactic reactions, drug allergies (e.g., allergies to penicillin, cephalosporins), eosinophilia-myalgia syndrome caused by ingestion of contaminated tryptophan, insect sting allergies; autoimmune diseases, such as rheumatoid arthritis, Graves' disease, Hughley's syndrome, psoriatic arthritis, multiple sclerosis, systemic lupus erythematosus, myasthenia gravis, immune thrombocytopenic purpura (ITP in adults, neonatal thrombocytopenic purpura, pediatric ITP), immune hemolytic anemia (autoimmune and drug-induced), Ivan's syndrome (platelet... And erythrocyte immune cytopenia), neonatal Rh disease, Cuban Stockholm syndrome (anti-GBM disease), celiac disease, autoimmune cardiomyopathy, juvenile-onset diabetes; glomerulonephritis, autoimmune thyroiditis, Behcet's disease; graft rejection (e.g. in transplantation), including allogeneic graft rejection or graft-versus-host disease; inflammatory bowel diseases, such as Crohn's disease and ulcerative colitis; spondyloarthritis; scleroderma; psoriasis (including T-cell mediated psoriasis) and inflammatory skin diseases, such as dermatitis, eczema, atopic dermatitis, allergic contact dermatitis, rubella; vasculitis (e.g. necrotizing, cutaneous, and allergic vasculitis); erythema nodosum; eosinophilia. Erythrombomyositis, 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 keratoconjunctivitis, scarring, anterior segment scarring, blepharitis, palpebral conjunctivitis, bullous diseases, cicatricial pemphigoid, conjunctival melanoma, papillary conjunctivitis, dry eye, episcleritis, glaucoma, glioma, granuloma annulare, Graves' ophthalmopathy, intraocular melanoma, pinguecula, proliferative vitreoretinopathy, pterygium, scleritis, uveitis, acute gout attack, gout or osteoarthritis.

[0196] (5) Pain, such as chronic idiopathic pain syndrome, neuropathic pain, hypoesthesia, tenderness to touch, migraine, toothache and postoperative pain.

[0197] (6) Depression, anxiety, diabetic neuropathy and bladder disorders, such as bladder outlet obstruction, overactive bladder, cystitis; myocardial reperfusion injury or cerebral ischemia injury.

[0198] In another aspect, the present invention relates to a compound of general formula 1, which is used to treat and / or prevent the diseases and symptoms mentioned above.

[0199] In another 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 diseases and symptoms mentioned above.

[0200] In another aspect of the invention, the invention relates to a method for treating or preventing the diseases and symptoms mentioned above, the method comprising administering to a human an effective amount of a compound of formula 1.

[0201] Combination therapy

[0202] The compounds of the present invention may be further combined with one or more, preferably one, additional therapeutic agents. According to one embodiment, the additional therapeutic agent is selected from the group consisting of: agents that can be used to treat the diseases or conditions described above, particularly those related to fibrotic diseases, inflammatory and immunomodulatory disorders, respiratory or gastrointestinal diseases or discomforts, inflammatory diseases or conditions of the joints or nasopharynx, eyes, and skin, such as cough, idiopathic pulmonary fibrosis, other interstitial lung diseases, asthma or allergic diseases, eosinophilic diseases, chronic obstructive pulmonary disease, atopic dermatitis, and diseases or conditions associated with autoimmune disorders (such as rheumatoid arthritis and atherosclerosis); or agents that can be used to treat ophthalmic diseases, pain, and depression.

[0203] Additional therapeutic agents suitable for such combinations specifically include, for example, therapeutic agents that enhance the therapeutic effect of one or more active substances on one of the mentioned indications and / or reduce the dosage of one or more active substances.

[0204] Therefore, the compounds of the present invention can 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.

[0205] Anti-fibrotic agents include, for example, nintedanib; pirfenidone; phosphodiesterase-IV (PDE4) inhibitors, such as roflumilast; autocrine motor factor 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; α-V / β-6 blocking inhibitors, such as BG-00011 / STX-100; recombinant permein-2 (PTX-2), such as PRM-151; c-Jun N-terminal kinase (JNK) inhibitors, such as CC-90001; galactoglobulin-3 inhibitors, such as TD-139; G-protein-coupled receptor 84 (GPR84) inhibitors, such as GLPG-1205; dual inhibitors of G-protein-coupled receptor 84 / G-protein-coupled receptor 40, such as PBI-4050; Rho-associated coiled-coil 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 domains of histyl-tRNA synthetase (HARS), such as ATYR-1923; prostaglandin synthase inhibitors, such as ZL-2102 / SAR-191801; 15 15-hydroxy-eicosapentaenoic acid (15-HEPE, e.g., DS-102); lysyl oxidase-like protein 2 (LOXL2) inhibitors, such as PAT-1251, PXS-5382 / PXS-5338; dual inhibitors of phosphatidylinositol 3-kinase (PI3K) / mammalian rapamycin target protein (mTOR), such as HEC-68498; calpain inhibitors, such as BLD-2660; mitogen-activated protein kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase 2 (MAPKAPK2) inhibitors, such as MMI-0100; transforming growth factor β1 (TGF-β1) small interfering RNA, such as TRK250 / BNC-1021; or lysophosphatidylcholine receptor antagonists, such as BMS-986278.

[0206] Cough suppressants include, for example, purine receptor 3 (P2X3) receptor antagonists, such as gefapixant, S-600918, BAY-1817080, or BLU-5937; neurokinin 1 (NK-1) receptor antagonists, such as orvepitant or aprepitant; nicotinic acetylcholine receptor α7 subunit stimulators, such as ATA-101 / bradanicline; codeine, gabapentin, pregablin, or azithromycin.

[0207] Anti-inflammatory agents include, for example, corticosteroids such as prednisolone or dexamethasone; and cyclooxygenase 2 (COX2) inhibitors such as celecoxib, rofecoxib, parecoxib, valdecoxib, deracoxib, etoricoxib, or lumiracoxib. ib); 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.

[0208] Anti-atopic dermatitis agents include, for example, cyclosporine, methotrexate, mycophenolate mofetil, azathioprine, phosphodiesterase inhibitors (e.g., apremilast, crisaborole), Janus-associated kinase (JAK) inhibitors (e.g., tofacitinib), neutralizing antibodies against IL-4 / IL-13 (e.g., dupilamab), neutralizing antibodies against IL-13 (e.g., lebrikizumab, tralokinumab), and neutralizing antibodies against IL-31 (nemolizumab).

[0209] Analgesics can be, for example, opioids such as morphine, oxymorphine, levopanol, oxycodone, propoxyphene, nalmefene, fentanyl, hydrocodone, hydromorphone, meripidine, methadone, nalorphine, naloxone, naltrexone, buprenorphine, butorphanol, nalbuphine, and pentazocine; or non-opioids such as acetophenamine.

[0210] Antidepressants include, for example, tricyclic antidepressants such as amitriptyline, clomipramine, despramine, doxepin, desipramine, imipramine, and nortriptyline; selective serotonin reuptake inhibitors (SSRIs) such as fluoxetine, paroxetine, sertraline, citalopram, and escitalopram; and norepinephrine reuptake inhibitors (SNRIs) such as maprotiline, lofepramine, mirtazapine, oxaprotiline, fezolamine, and tomoxetine. Antidepressants such as mianserin, buproprion, hydroxybuproprion, nomifensine, and viloxazine; serotonin-norepinephrine dual reuptake inhibitors (SNRIs), such as duloxetine, venlafaxine, desvenlafaxine, and levomilnacipran; atypical antidepressants such as trazodone, mirtazapine, vortioxetine, vilazodone, and bupropion; or monoamine oxidase inhibitors (MAOIs), such as tranylcypromine, phenelzine, or isocarboxazid.

[0211] Anxiolytics are, for example, benzodiazepines, such as alprazolam, bromazepam, chlordiazepoxide, clonazepam, clonazepam, diazepam, flurazepam, lorazepam, oxazepam, temazepam, triazolam, or tofisopam; or non-benzodiazepines. Phenotypic hypnotics, such as eszopiclone, zaleplon, zolpidem, or zopiclone; or carbamates, such as meprobamate, carisoprodol, tybamate, or lorbamate; or antihistamines, such as hydroxyzine, chlorpheniramine, or diphenhydramine.

[0212] The sedatives are, for example, barbiturate sedatives such as amobarbital, aprobarbital, butabarbital, butabital, mephobarbital, metharbital, methoexital, pentobarbital, secobarbital, talbutal, theamylal, or thiopental; or non-barbiturate sedatives such as glutethimide, meprabarbital, methaqualone, or dichloal phenazone.

[0213] Skeletal muscle relaxants include, for example, baclofen, mepparidine, callisopredo, cyclobenzaprine, metaxalone, mesocarbamol, tizanidine, chlorzoxazone, or orphenadrine.

[0214] Other suitable combination therapies include the following inhibitors: acetylcholinesterase inhibitors, such as donepezil; 5-HT-3 antagonists, such as ondansetron; metabolite glutamate receptor antagonists; antiarrhythmic drugs, such as mexiletine or phenytoin; or NMDA receptor antagonists.

[0215] Other suitable combinations include incontinence drugs, such as anticholinergic agents like oxybutynin, tolterodine, darifenacin, fesoterodine, solifenacin, or trospium; or bladder muscle relaxants like mirabegron; or alpha blockers like tamsulosin, alfuzosin, silodosin, doxazosin, or terazosin.

[0216] The dosage of the above combination of drugs is usually 1 / 5 to 1 / 1 of the usually recommended minimum dose.

[0217] Therefore, in another aspect, the present invention relates to the use of compounds according to the invention in combination with one or more additional therapeutic agents as described in the context for the treatment of diseases or conditions that may be affected by or mediated by TRPA1, particularly diseases or conditions as described in the context.

[0218] In another aspect, the present invention relates to a method for treating a patient with a disease or symptom that may be affected by TRPA1 inhibition, comprising the steps of administering to a patient requiring such treatment a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, and a therapeutically effective amount of one or more additional therapeutic agents.

[0219] In another aspect, the present invention relates to the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in combination with one or more additional therapeutic agents for the treatment of a disease or condition in patients in need that may be affected by TRPA1 inhibition.

[0220] In another aspect, the present invention relates to a method for treating a patient with a disease or symptom mediated by TRPA1 activity, comprising the steps of administering to a patient (preferably a human) requiring such treatment a therapeutically effective amount of the compound of the present invention and a therapeutically effective amount of one or more additional therapeutic agents as described in the context.

[0221] The use of the compound according to the invention in combination with this additional therapeutic agent can be performed simultaneously or at staggered times.

[0222] The compounds and one or more additional therapeutic agents according to the invention may coexist in a formulation, such as a tablet or capsule, or may be present separately in two identical or different formulations, such as in the form of a kit containing so-called aliquots.

[0223] Therefore, 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 in the context, optionally together with one or more inert carriers and / or diluents.

[0224] In another aspect, the present invention relates to the use of compounds according to the invention in a cough measuring device.

[0225] Other features and advantages of the invention will become apparent from the following more detailed examples, which illustrate the principles of the invention by way of example.

[0226] preparation

[0227] The compounds and intermediates according to the invention 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 explained more fully below and particularly as described in the experimental section. In some cases, the order of the reaction steps may be varied. Variations of reaction methods known to those skilled in the art but not described in detail herein may also be used.

[0228] The general methods for preparing the compounds according to the invention will become apparent to those skilled in the art who study the following processes. Conventional protecting groups can be used to protect any functional groups in the starting material or intermediates. These protecting groups can then be cleaved at appropriate stages within the reaction sequence using methods familiar to those skilled in the art.

[0229] The compounds according to the invention are prepared by the synthetic methods described below, wherein the substituents of each general formula have the meanings given above. These methods are intended to be illustrative of the invention and not to limit its subject matter or the scope of the compounds claimed in these examples. When the preparation of a starting compound is not described, it is commercially available or can be prepared similarly to the known compounds or methods described herein. Substances described in the literature are prepared according to disclosed synthetic methods. Abbreviations are as defined in the Examples section.

[0230] Process 1:

[0231]

[0232] In process 1, chloromethyltetrazole is N-alkylated in the presence of a base (e.g., K₂CO₃) with a suitable acetone derivative carrying a leaving group “LG” (e.g., Cl or Br) at the carbonyl α-position, yielding a mixture of two regiomeric isomers. Undesired regiomeric isomers (not shown) can be removed chromatographically using an appropriate gradient. The resulting ketone (A) can be reduced in a mirror-selective manner using a suitable catalytic system with a combination of a transition metal complex (e.g., Ru or Ir) and a chiral ligand (e.g., [(1S,2S)-2-amino-1,2-diphenylethyl](4-toluenesulfonyl)amide) and a hydrogen source (such as a triethylamine formate complex) to give an alcohol (B). The final compound (F) can be synthesized by alkylating 6-methyl-4-oxo-3H,4H-furano[2,3-d]pyrimidine-5-carboxylic acid (C) with intermediate (B) in the presence of a base (such as K2CO3), and subsequently amidating the carboxylic acid (D) with a coupling agent (such as HATU) and an ammonia source (such as ammonium carbonate) in the presence of a base. Alternatively, the final compound (F) can be synthesized by amidating (C) with a coupling agent (such as CDI) and ammonia, followed by alkylating the amide (E) with intermediate (B).

[0233] Process 2:

[0234]

[0235] In procedure 2, the synthesis of the final compound (K) is described. 1-(4,6-dichloropyrimidin-5-yl)ethyl-1-one undergoes nucleophilic aromatic substitution with ethyl glycolate in the presence of a base (such as potassium tert-butoxide) to give intermediate (G), which is subsequently treated with sodium ethoxide and a base (such as potassium tert-butoxide) to give intermediate (H). The protecting group of the pyrimidinone in (H) is removed in acetonitrile with, for example, sodium iodide and trimethylsilane chloride to give ester (I), which can be reacted with ammonia in the presence of a Lewis acid (such as calcium chloride) to give amide (J). Finally, amide (J) is alkylated with intermediate (B) in the presence of a base (such as potassium carbonate) to give the final compound (K).

[0236] Example

[0237] preparation

[0238] The compounds and intermediates according to the invention 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 in a manner similar to the preparation methods explained more fully below and particularly as described in the Experimental Section. In some cases, the order of the reaction process may vary. Variations of these reactions known to those skilled in the art but not described in detail herein may also be used. Those skilled in the art will find the general method for preparing the compounds according to the invention apparent upon studying the following process. The starting compounds are commercially available or can be prepared by the methods described in the literature or herein, or can be prepared in a similar or analogous manner. Prior to the reaction, any corresponding functional groups in the starting compounds may be protected with conventional protecting groups. These protecting groups can then be cleaved at appropriate stages within the reaction sequence using methods familiar to those skilled in the art and described in the literature, such as “Protecting Groups”, 3rd edition, Philip J. Kocienski, Thieme, 2005 and “Protective Groups in Organic Synthesis”, 4th edition, Peter GMWuts, Theodora W. Greene, John Wiley & Sons, 2006. The terms “ambient temperature” and “room temperature” are used interchangeably and represent approximately 20°C, for example, a temperature between 19 and 24°C.

[0239] abbreviation:

[0240]

[0241]

[0242]

[0243] Preparation of intermediates

[0244] Intermediate I

[0245] Intermediate I.1 (General Procedure)

[0246] 2-[5-(chloromethyl)-2H-1,2,3,4-tetrazol-2-yl]-1-(4-chlorophenyl)ethyl-1-one

[0247]

[0248] At room temperature, with stirring, 1.63 g (11.8 mmol) of K₂CO₃ was added to 15 mL of DMA containing 1.00 g (8.44 mmol) of 5-(chloromethyl)-2H-1,2,3,4-tetraazole and 2.17 g (9.28 mmol) of 4-chlorobenzoylmethyl bromide. The reaction mixture was stirred at room temperature for 30 minutes and then filtered. The filtrate was diluted with water and saturated NaCl aqueous solution and extracted three times with EtOAc. The combined organic phases were washed with water, dried over Na₂SO₄, filtered through activated carbon, and the solvent was removed under reduced pressure. The residue was purified by column chromatography (silica gel; CH / EtOAc, 80 / 20 to 50 / 50 gradient) to give the product.

[0249] C 10 H8Cl2N4O (M=271.1g / mol)

[0250] ESI-MS: 271 [M+H] +

[0251] R t (HPLC): 1.01 min (Method B)

[0252] The following compounds were prepared using a procedure similar to that described for intermediate I.1, with appropriate starting materials. Those skilled in the art will understand that these similar examples may involve variations in general reaction conditions.

[0253]

[0254]

[0255] Intermediate II

[0256] Intermediate II.1 (General Procedure) (1R)-2-[5-(chloromethyl)-2H-1,2,3,4-tetrazol-2-yl]-1-(4-chlorophenyl)ethyl-1-ol

[0257]

[0258] Under an inert atmosphere, 1.30 g (4.80 mmol) of 1-(4-chlorophenyl)-2-[5-(chloromethyl)-2H-1,2,3,4-tetraazol-2-yl]ethyl-1-one (intermediate I.1) was dissolved in 20 mL of ACN. 12 mg (0.02 mmol) of chloro([(1S,2S)-2-amino-1,2-diphenylethyl](4-toluenesulfonyl)amido(mesene)ruthenium(II) (CAS 174813-81-1) was added, followed by dropwise addition of 0.72 mL (1.73 mmol) of triethylamine formate complex (5:2). After stirring at room temperature for 3 hours, the solvent was removed under reduced pressure. Water was added to the remaining crude mixture, and the mixture was extracted with EtOAc. The organic layers were combined, dried over Na2SO4, filtered, treated with activated carbon, filtered again, and the solvent was removed under reduced pressure to give intermediate II.1.

[0259] C 10 H 10 Cl2N4O (M=273.1g / mol)

[0260] ESI-MS: 273 [M+H] +

[0261] R t (HPLC): 0.96 min (Method B)

[0262] The following compounds were prepared using a procedure similar to that described for intermediate II.1, with appropriate starting materials. Those skilled in the art will understand that these similar examples may involve variations in general reaction conditions.

[0263]

[0264] Intermediate III

[0265] Intermediate III.1

[0266] 1-(7-fluoro-1-benzofuran-2-yl)ethyl-1-one

[0267]

[0268] A stirred solution of 6.00 g (42.8 mmol) of 3-fluoro-2-hydroxybenzaldehyde in 60 mL of acetone was cooled to 0 °C and subsequently treated with 9.47 g (68.5 mmol) of potassium carbonate. After stirring at 0 °C for 10 min, 5.12 mL (64.2 mmol) of chloroacetone was added dropwise, and the reaction mixture was stirred at 90 °C for 1 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude residue was extracted with EtOAc / water, and the organic phase was concentrated under reduced pressure to give intermediate III.1.

[0269] C 10 H7FO2 (M = 178.2 g / mol)

[0270] ESI-MS: 179 [M+H] +

[0271] R t (HPLC): 0.50 min (Method A)

[0272] The following compounds were prepared in a manner similar to intermediate III.1. Those skilled in the art will understand that variations in general reaction conditions may be involved in these similar examples.

[0273]

[0274] Intermediate IV

[0275] Intermediate IV.1

[0276] 2-Bromo-1-(7-fluoro-1-benzofuran-2-yl)ethyl-1-one

[0277]

[0278] At room temperature, with stirring, 14.81 g (30.7 mmol) of tetrabutylammonium tribromide in 3.3 mL MeOH and 32 mL THF was added dropwise to 5.47 g (30.7 mmol) of 1-(7-fluoro-1-benzofuran-2-yl)ethyl-1-one (intermediate III.1) in 66 mL THF. The reaction mixture was stirred at room temperature for 2 hours, concentrated under reduced pressure, and the residue was extracted with EtOAc / water. The organic layer was concentrated under reduced pressure, and the crude material was purified by column chromatography (silica gel; hexane / EtOAc, gradient).

[0279] C 10 H6BrFO2 (M=257.1g / mol)

[0280] ESI-MS: 257 / 259 [M+H] +

[0281] R t (HPLC): 0.58 min (Method A)

[0282] The following compounds were prepared in a manner similar to intermediate IV.1. Those skilled in the art will understand that variations in general reaction conditions may be involved in these similar examples.

[0283]

[0284]

[0285] intermediate V

[0286] 6-Methyl-4-oxo-3H,4H-furano[2,3-d]pyrimidine-5-carboxamide

[0287]

[0288] At room temperature, 2.90 g (14.94 mmol) of 6-methyl-4-oxo-3H,4H-furano[2,3-d]pyrimidin-5-carboxylic acid (CAS: 852399-94-1, European Journal of Medicinal Chemistry, 2018, Vol. 144, pp. 330-348) and 2.66 g (16.43 mmol) of CDI were stirred in 60 mL of THF for 18 hours. Subsequently, 90 mL of NH3 (0.5 mol / L) in THF was added and stirring continued at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure and poured into an aqueous solution of NaHCO3 (100 mL water + 40 mL saturated NaHCO3 solution). The resulting precipitate was filtered off and dried to give intermediate V.

[0289] C8H7N3O3 (M=193.2g / mol)

[0290] ESI-MS: 194 [M+H] +

[0291] R t (HPLC): 0.61 min (Method B)

[0292] Intermediate VI

[0293] 2-[(5-acetyl-6-chloropyrimidin-4-yl)oxy]ethyl acetate

[0294]

[0295] At 0 °C, potassium tert-butoxide solution (1.26 mL, 1.0 mol / L) was slowly added to 111 μL (1.15 mmol) of ethyl glycolate in 5.0 mL of THF with stirring. After stirring at 0 °C for 35 min, 200 mg (1.05 mmol) of 1-(4,6-dichloropyrimidin-5-yl)ethyl-1-one was added, and stirring was continued at 0 °C for 2 h, followed by stirring at room temperature for 3 h. The reaction mixture was concentrated under reduced pressure and purified by column chromatography (silica gel; hexane / EtOAc, gradient).

[0296] C 10 H 11 ClN2O4 (M=258.66g / mol)

[0297] ESI-MS: 259 [M+H] +

[0298] R t (HPLC): 1.05 min (Method B)

[0299] Intermediate VII

[0300] 4-Ethoxy-5-methylfurano[2,3-d]pyrimidine-6-carboxylic acid ethyl ester

[0301]

[0302] At -12°C, a solution of sodium ethoxide in ethanol (2.95 mL, 21%) was slowly added to a stirred solution of intermediate VI in THF. The reaction mixture was stirred at 0°C for 90 min, and a solution of potassium tert-butoxide in THF (4.31 mL, 1.0 mol / L) was added, with stirring continued overnight at room temperature. Then, potassium tert-butoxide in THF (1.0 mL, 1.0 mol / L) was added, and stirring continued for 3 h at room temperature. The reaction mixture was acidified with acetic acid and concentrated under reduced pressure. The reaction mixture was extracted with water / EtOAc, and the organic extract was dried over MgSO4, filtered, and concentrated under reduced pressure. Intermediate VII was purified by reversed-phase HPLC (ACN / H2O gradient, 0.1% TFA).

[0303] C 12 H 14 N₂O₄ (M = 250.25 g / mol)

[0304] ESI-MS: 251 [M+H] +

[0305] R t (HPLC): 0.63 min (Method A)

[0306] Intermediate VIII

[0307] ethyl 5-methyl-4-oxo-3H,4H-furano[2,3-d]pyrimidine-6-carboxylate

[0308]

[0309] Add 90 mg (0.60 mmol) of sodium iodide to 50 mg (0.20 mmol) of intermediate VII in 2 mL of ACN and stir the mixture for 3 minutes at room temperature. Then, add 76 μL (0.60 mmol) of trimethylsilane chloride, seal the reaction vessel, and continue stirring at room temperature for 2 hours. Quench the reaction mixture with 10 mL of water and stir for 5 minutes at room temperature. Filter out the precipitate, wash with water, and dry under reduced pressure at 50 °C to give intermediate VIII.

[0310] C 10 H 10 N₂O₄ (M = 222.20 g / mol)

[0311] ESI-MS: 223 [M+H] +

[0312] R t (HPLC): 0.78 min (Method B)

[0313] Intermediate IX

[0314] 5-Methyl-4-oxo-3H,4H-furano[2,3-d]pyrimidine-6-carboxamide

[0315]

[0316] In a sealed container, a mixture of 800 mg (3.60 mmol) of intermediate VIII and 400 mg (3.60 mmol) of CaCl2 in 128 mL of ammonia / MeOH (7 mol / L) was stirred at 50 °C for 36 hours. The reaction mixture was concentrated under reduced pressure, stirred in 50 mL of water, and the resulting precipitate was filtered off. The precipitate was washed with water and dried under reduced pressure at 60 °C to give intermediate IX.

[0317] C8H7N3O3 (M=193.16g / mol)

[0318] ESI-MS: 194 [M+H] +

[0319] R t (HPLC): 0.50 min (Method H)

[0320] Intermediate X

[0321] Intermediate X.1

[0322]

[0323] A mixture of 120 mg (0.47 mmol) of 6-methyl-4-oxo-3H,4H-furano[2,3-d]pyrimidin-5-carboxylic acid (CAS: 852399-94-1), 92 mg (0.47 mmol) of (1R)-2-[5-(chloromethyl)-2H-1,2,3,4-tetraazol-2-yl]-1-(4-methylphenyl)ethyl-1-ol (intermediate II.9), and 98 mg (0.71 mmol) of K₂CO₃ in 2 mL of DMA was stirred overnight at room temperature. Subsequently, 98 mg (0.71 mmol) of K₂CO₃ was added, and the mixture was stirred at 60 °C for 2 h, followed by stirring at 80 °C for 5 h. The reaction mixture was cooled to room temperature and purified by reversed-phase HPLC (ACN / H₂O gradient, 0.1% TFA) to obtain the desired product.

[0324] C 19 H 18 N6O5 (M=410.38g / mol)

[0325] ESI-MS: 411 [M+H] +

[0326] R t (HPLC): 0.90 min (Method H)

[0327] The following compounds were prepared using a procedure similar to that described for intermediate X.1, with appropriate starting materials. Those skilled in the art will understand that these similar examples may involve variations in general reaction conditions.

[0328]

[0329] Preparation of the final compound

[0330] Example 1 (General Procedure A)

[0331] 3-({2-[(2R)-2-hydroxy-2-(4-methylphenyl)ethyl]-2H-1,2,3,4-tetrazo-5-yl}methyl)-6-methyl-4-oxo-3H,4H-furano[2,3-d]pyrimidine-5-carboxamide

[0332]

[0333] At room temperature, a mixture of 1.50 g (7.77 mmol) 6-methyl-4-oxo-3H,4H-furano[2,3-d]pyrimidin-5-carboxamide (intermediate V), 3.22 g (23.30 mmol) K₂CO₃, and 1.96 g (7.77 mmol) (1R)-2-[5-(chloromethyl)-2H-1,2,3,4-tetraazol-2-yl]-1-(4-methylphenyl)ethyl-1-ol (intermediate II.9) in 25 mL DMA was stirred overnight. The reaction mixture was poured over ice water and extracted three times with EtOAc. The combined organic layers were stirred with MgSO₄, filtered, concentrated under reduced pressure, and purified by reversed-phase HPLC (ACN / H₂O gradient, 0.3% TFA) to obtain the desired product.

[0334] C 19 H 19 N7O4 (M=409.4g / mol)

[0335] ESI-MS: 410 [M+H] +

[0336] R t (HPLC): 0.73 min (Method B)

[0337] 1 H NMR (400MHz, DMSO-d6) δppm: 2.25 (s, 3H), 2.72 (s, 3H), 4.74 (d, J = 6.6Hz, 2H), 5.02-5.08 (m,1H),5.58(s,2H),5.74(s,1H),7.10(d,J=8.0Hz,2H),7.21(d,J=8.0Hz,2H),7.55(br d,J=1.3Hz,1H),8.74(s,1H),9.16-9.22(m,1H)

[0338] The following compounds were prepared using a procedure similar to that described for General Procedure A of Example 1, with appropriate starting materials. Those skilled in the art will understand that these similar examples may involve variations in general reaction conditions.

[0339]

[0340]

[0341] The analytical data for the compounds are described in the table above:

[0342]

[0343]

[0344]

[0345] Preparation of the final compound

[0346] Example 1 (General Procedure B)

[0347] 3-({2-[(2R)-2-hydroxy-2-(4-methylphenyl)ethyl]-2H-1,2,3,4-tetrazo-5-yl}methyl)-6-methyl-4-oxo-3H,4H-furano[2,3-d]pyrimidine-5-carboxamide

[0348]

[0349] Add 32 mg (0.09 mmol) of HATU and 73 μL (0.43 mmol) of DIPEA to 35 mg (0.09 mmol) of 3-({2-[(2R)-2-hydroxy-2-(4-methylphenyl)ethyl]-2H-1,2,3,4-tetraazol-5-yl}methyl)-6-methyl-4-oxo-3H,4H-furano[2,3-d]pyrimidin-5-carboxylic acid (intermediate X.1) at room temperature while stirring. After 5 minutes, add 82 mg (0.85 mmol) of ammonium carbonate and stir the mixture overnight at room temperature. Purify the mixture by reversed-phase HPLC (ACN / H2O gradient, 0.1% NH3) to obtain the desired product.

[0350] The following compounds were prepared using a procedure similar to that described for General Procedure B of Example 1, with appropriate starting materials. Those skilled in the art will understand that these similar examples may involve variations in general reaction conditions.

[0351]

[0352]

[0353] The analytical data for the compounds are described in the table above:

[0354]

[0355] Analytical HPLC methods

[0356] Method A

[0357]

[0358] Analytical column: XBridge BEH (Waters) C18 2.1 × 30 mm 1.7 μm; column temperature: 60℃

[0359] Method B

[0360]

[0361]

[0362] Analytical column: Stable Bond (Agilent) C18 3.0×30mm 1.8μm; Column temperature: 60℃

[0363] Method C

[0364]

[0365] Analytical column: Xbridge (Waters) C18 3.0 × 30 mm 2.5 μm; Column temperature: 60℃

[0366] Method D

[0367]

[0368] Analytical column: XBridge C18 3.0×30mm 2.5μm (Waters); Column temperature: 60℃

[0369] Method E

[0370]

[0371] Analytical column: Sunfire (Waters); C18 3.0 × 30 mm 2.5 μm, column temperature: 60℃

[0372] Method F

[0373]

[0374] Analytical column: XBridge BEH (Waters) C18 2.1 × 30 mm 2.5 μm; Column temperature: 60℃

[0375] Method G

[0376]

[0377] Analytical column: Zorbax StableBond C18 (Agilent) 1.8 μm; 2.1 × 30 mm; column temperature: 60 °C

[0378] Method H

[0379]

[0380]

[0381] Analytical column: Sunfire (Waters) 2.5 μm; 3.0 × 30 mm; column temperature: 60℃

[0382] Method I

[0383]

[0384] Analytical column: Sunfire C18 (Waters) 2.5 μm; 3.0 × 30 mm; column temperature: 60℃

[0385] Method J

[0386]

[0387] Analytical column: Acquity UPLC BEH; C8 2.1 × 150 mm 1.7 μm; column temperature: 55℃

[0388] Method K

[0389]

[0390] Analytical column: XBridge BEH Phenyl (Waters) 2.1 × 30 mm ≤ 1.7 μm; Column temperature: 60 °C

Claims

1. A compound according to formula (I) Where A is selected from and R 1 It is H3C or H2N(O)C; and R 2 It is H3C or H2N(O)C; Its limitations are: When R 1 When it is H2N(O)C, R 2 For H3C; When R 2 When it is H2N(O)C, R 1 It is H3C.

2. The compound of formula (I) according to claim 1, wherein the compound is selected from the group consisting of: and 3. The compound of formula (I) according to claim 1, wherein the compound is selected from the group consisting of: and 4. A compound with the following formula:

5. A compound of the following formula:

6. A compound with the following formula:

7. A compound with the following formula:

8. A compound with the following formula:

9. A compound with the following formula:

10. A compound with the following formula:

11. A compound with the following formula:

12. A compound with the following formula:

13. A compound with the following formula:

14. A salt of a compound according to any one of claims 1 to 13.

15. A pharmaceutically acceptable salt of a compound according to any one of claims 1 to 13.

16. A pharmaceutical composition comprising at least one compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients.

17. Use of a compound of any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof in the preparation of a medicament.

18. Use of a compound of any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment or prevention of inflammatory airway diseases or fibrotic diseases or cough.

19. Use of a compound of any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment or prevention of idiopathic lung disease (IPF) or cough.

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