Tetrazolium derivatives as TRPA1 inhibitors

Abstract

The present invention provides certain tetrazole derivatives that are inhibitors of transient receptor potential ankyrin 1 (TRPA1) and are therefore useful in treating diseases treatable by inhibiting TRPA1. The present invention also provides pharmaceutical compositions containing the same and methods for preparing the compounds.

Description

Technical Field

[0001] The present invention provides certain tetrazole derivatives that are inhibitors of transient receptor potential ankyrin 1 (TRPA1) and are therefore useful in treating diseases treatable by inhibiting TRPA1. The present invention also provides pharmaceutical compositions containing the same and methods for preparing the compounds. Background Art

[0002] Transient receptor potential channels (TRP channels) are a group of voltage-gated ion channels located primarily on the plasma membrane of numerous mammalian cell types. There are approximately 30 structurally related TRP channels, which are classified into the following categories: 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), which give rise to the "A" designation for ankyrin (Montell, 2005).

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

[0004] TRPA1 is best known as a sensor for environmental stimuli that generate somatosensory modalities such as pain, cold, and itch. TRPA1 is activated by a variety of reactive, electrophilic stimuli (e.g., allyl isothiocyanate, reactive oxygenates), as well as non-reactive compounds (e.g., icilin), and is implicated in cough associated with asthma, chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis (IPF), or post-viral cough, or chronic idiopathic cough and cough in sensitive patients (Song and Chang, 2015; Grace and Belvisi, 2011). Based on studies showing that cough induces elevated TGF-β (Xie et al., 2009; Froese et al., 2016; Tschumperlin et al., 2003; Yamamoto et al., 2002; Ahamed et al., 2008), TRPA1 inhibitors are suitable for treating IPF, where cough is very common due to the link between cough and lung damage. TRPA1 antagonists inhibit calcium signaling triggered 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 have been effective in studies 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-associated itch (Cevikbas et al., 2014). Gain of function of human TRPA1 is associated with familial paroxysmal pain syndrome (Kremeyer et al., 2010). TRPA1 antagonists are also effective in a behavioral model of migraine-related allodynia (Edelmayer et al., 2012). When compared to TRPA1 expression in trigeminal ganglia innervating healthy teeth, TRPA1 is selectively increased in trigeminal ganglia innervating damaged 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 to alleviate postoperative pain. TRPA1 knockout mice and wild-type mice exhibit anxiolytic and antidepressant-like phenotypes after treatment with TRPA1 antagonists (de Moura et al., 2014). Based on studies demonstrating a mechanistic link between AMPK and TRPA1, TRPA1 inhibitors are expected to be beneficial in the treatment of diabetic neuropathy (Hiyama et al., 2018; Koivisto and Pertovaara, 2013; Wang et al., 2018). TRPA1 knockout mice have been shown to have smaller myocardial infarct sizes compared to wild-type mice (Conklin et al., 2019).TRPA1 gene ablation and pharmacological intervention inhibit TNBS-induced colitis in mice (Engel et al., 2011). In a mouse model of cerebral ischemia, TRPA1 gene ablation and a TRPA1 antagonist reduce myelin damage (Hamilton et al., 2016). In a monosodium urate mouse model of gout, urate crystals and joint inflammation are reduced in TRPA1 knockout mice (Moilanen et al., 2015). TRPA1 deficiency in rats improves joint inflammation and hyperalgesia in a rat model of acute gout (Trevisan et al., 2014). TRPA1 activation triggers an inflammatory response in osteoarthritic chondrocytes (Nummenmaaet et al., 2016). TRPA1 inhibition and gene ablation reduce inflammatory mediators in chondrocytes and murine cartilage in osteoarthritic mice (Nummenmaa et al., 2016). Finally, TRPA1 knockout mice demonstrate improved weight-bearing in the osteoarthritic limb in a model of MIA-induced knee swelling (Horvath et al., 2016). TRPA1 is differentially expressed in the bladder epithelium of rats (Du et al., 2007) and patients with bladder outlet obstruction (Du et al., 2008). TRPA1 receptor modulation alleviates overactive bladder in a rat model of spinal cord injury (Andrade et al., 2011), and intrathecal administration of a TRPA1 antagonist alleviates cyclophosphamide-induced cystitis in rats with hyperreflexic micturition (Chen et al., 2016).

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

[0006] Various structural classes of TRPA1 inhibitors are reviewed in S. Skerratt, Progress in Medicinal Chemistry, 2017, Vol. 56, 81-115 and D. Preti, G. Saponaro, A. Szallasi, Pharm. Pat. Anal. (2015) 4(2), 75-94.

[0007] WO2017 / 060488 discloses compounds as TRPA1 antagonists having the following general structural formula

[0008]

[0009] However, there is no disclosure of TRPA1 activity of Examples 28 and 29 containing a tetrazolyl ring.

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

[0011]

[0012] Compound 31, wherein R is OH, was disclosed as having an IC in FLIPR analysis. 50 It antagonizes TRPA1 activity at 58 nM and has an intrinsic clearance of <14 μL / min / kg in human liver microsomes. DETAILED DESCRIPTION

[0013] The present invention discloses novel tetrazole derivatives which are inhibitors of transient receptor potential ankyrin 1 (TRPA1) having suitable pharmacological and pharmacokinetic properties, enabling their use as agents for treating conditions and / or diseases that can be treated by inhibiting TRPA1.

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

[0015] Compounds of the present invention

[0016] The present invention provides novel tetrazole derivatives that are unexpectedly potent inhibitors of TRPA1 (Assay A) and are further characterized by

[0017] Improved stability in human liver microsomes (Assay B)

[0018] Stability was improved in human hepatocytes (Assay C).

[0019] The compounds of the present invention differ structurally from Examples 28 and 29 in WO2017 / 060488 in that they contain a monocyclic dioxodiprimidine core with an N-substituent, an amide substituent, and a substituent adjacent to a secondary aliphatic alcohol. They also differ structurally from Example 31 in L. Schenkel et al., J. Med. Chem. 2016, 59, 2794-2809 in that they contain a tetrazolyl ring. These structural differences unexpectedly result in 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 those disclosed in the prior art in terms of the combination of the following parameters:

[0021] - Potency as a TRPA1 inhibitor

[0022] - Stability in human liver microsomes

[0023] -Stability in human hepatocytes.

[0024] Stability in human liver microsomes refers to the susceptibility of a compound to biotransformation in the case of selecting and / or designing a drug with favorable pharmacokinetic properties as the first screening step. The liver is the main metabolic site of many drugs. Human liver microsomes contain cytochrome P450 (CYP) and therefore represent a model system for studying in vitro phase I drug metabolism. The enhanced stability in human liver microsomes is associated with several advantages (including increased bioavailability and an appropriate half-life), which can achieve a reduced dosage and lower frequency for patients. Therefore, the enhanced stability in human liver microsomes is a favorable feature of compounds to be used for drugs. Therefore, in addition to being able to inhibit TRPA1, the compounds of the present invention are also expected to have favorable in vivo clearance and therefore have a desired duration of action in humans.

[0025] Stability in human hepatocytes refers to the susceptibility of a compound to biotransformation in the context of selecting and / or designing a drug with favorable pharmacokinetic properties. The liver is the primary metabolic site 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 liver-specific transporter-mediated processes, and therefore represents a more complete system for drug metabolism research). The enhanced stability in human hepatocytes is associated with several advantages (including increased bioavailability and an appropriate half-life), which can achieve a reduction in the patient's dosage and a lower frequency. Therefore, the enhanced stability in human hepatocytes is a favorable feature of compounds to be used for drugs.

[0026] The present invention provides novel compounds according to formula (I)

[0027]

[0028] in

[0029] A is selected from the group consisting of phenyl, thienyl, benzothienyl or benzofuranyl, which are unsubstituted or substituted by halogen, C 1-4 Alkyl, C 1-4 Fluoroalkyl, C 3-4 Cycloalkyl, C 3-4 Cyclofluoroalkyl, -OC 1-4 Alkyl, -O-cyclopropyl and NC-group R 3 one, two or three members of the

[0030] or

[0031] A is selected from the group consisting of:

[0032]

[0033] R 1 Choose from C 1-4 Alkyl, C 1-4 Fluoroalkyl, C 3-6 Cycloalkyl, R 4 -(H2C) m - and R 5 -(H2C) n - the group formed;

[0034] in

[0035] m is 1 or 2;

[0036] n is 2;

[0037] R 4 C 3-6 Cycloalkyl;

[0038] R 5 For-OC 1-4 Alkyl or -OC 1-4 Fluoroalkyl;

[0039] R 2 Choose from H, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 3-6 Cyclofluoroalkyl, HO-C 1-4 Alkyl-, C 1-4 Fluoroalkyl, R 6 -(H2C) p -、R 7 -(H2C) q -、R 6 -(H(R 8 )C) p - and R 7 -(H(R 9 )C) q - the group formed;

[0040] in

[0041] p is 1 or 2;

[0042] q is 2;

[0043] R 6 Choose from HO-C 1-2 Alkyl-, C 3-6 the group consisting of cycloalkyl, C-morpholinyl, C-imidazolyl and C-pyrazolyl;

[0044] wherein the C-pyrazolyl, C-imidazolyl and C-morpholinyl are unsubstituted or C 1-4 Alkyl or C 1-4 Fluoroalkyl substitution;

[0045] R 7 Choose Free-OC 1-4 Alkyl, -OC 1-4 Fluoroalkyl, C 1-4 the group consisting of alkyl-S(O)2-, N-morpholinyl, N-imidazolyl and N-pyrazolyl;

[0046] wherein the N-pyrazolyl, N-imidazolyl, N-morpholinyl is unsubstituted or C 1-4 Alkyl or C 1-4 Fluoroalkyl substitution;

[0047] R 8 and R 9 are independently selected from H or C 1-4 alkyl.

[0048] Another embodiment of the present invention relates to a compound of formula (I), wherein

[0049] A is selected from the group consisting of phenyl, thienyl, benzothienyl or benzofuranyl, which are unsubstituted or substituted by halogen, C 1-4 Alkyl, -OC 1-4 Alkyl and NC-group R 3 One or two members are replaced;

[0050] or

[0051] A is

[0052]

[0053] R 1 Choose from C 1-4 Alkyl, C 3-6 Cycloalkyl, R 4 -(H2C) m - and R 5 -(H2C) n - the group formed;

[0054] in

[0055] m is 1 or 2;

[0056] n is 2;

[0057] R 4 C 3-6 Cycloalkyl;

[0058] R 5 For-OC 1-4alkyl;

[0059] R 2 Choose from H, C 1-4 Alkyl, C 3-6 Cycloalkyl, HO-C 1-4 Alkyl-, C 1-4 Fluoroalkyl, R 6 -(H2C) p - and R 7 -(H2C) q - the group formed;

[0060] in

[0061] p is 1 or 2;

[0062] q is 2;

[0063] R 6 Choose from C 3-6 the group consisting of cycloalkyl, C-morpholinyl, C-imidazolyl and C-pyrazolyl;

[0064] wherein the C-pyrazolyl, C-imidazolyl and C-morpholinyl are unsubstituted or C 1-4 Alkyl substitution;

[0065] R 7 Choose Free-OC 1-4 Alkyl, -OC 1-4 Fluoroalkyl, C 1-4 the group consisting of alkyl-S(O)2-, N-morpholinyl, N-imidazolyl and N-pyrazolyl;

[0066] wherein the N-pyrazolyl, N-imidazolyl, N-morpholinyl is unsubstituted or C 1-4 Alkyl substitution.

[0067] Another embodiment of the present invention relates to a compound of formula (I)

[0068] in

[0069] A is selected from the group consisting of phenyl, thienyl, benzothienyl or benzofuranyl, which is unsubstituted or substituted by a group R consisting of Cl, F, Br, H3C, H3C-O- and NC- 3 One or two members are replaced;

[0070] or

[0071] A is

[0072]

[0073] And the substituent R 1 and R 2 As defined in the preceding embodiments.

[0074] Another embodiment of the present invention relates to a compound of formula (I)

[0075] in

[0076] A is selected from the group consisting of:

[0077]

[0078] and

[0079]

[0080] The group R is unsubstituted or composed of Cl, F, Br, H3C, H3C-O- and NC- 3 One or two members are replaced,

[0081] or

[0082] A is

[0083]

[0084] And the substituent R 1 and R 2 As defined in any of the preceding embodiments.

[0085] Another embodiment of the present invention relates to a compound of formula (I), wherein

[0086] A is selected from the group consisting of:

[0087]

[0088]

[0089]

[0090] and

[0091]

[0092] And the substituent R 1 and R 2 As defined in any of the preceding embodiments.

[0093] Another embodiment of the present invention relates to a compound of formula (I), wherein

[0094] R 1 Choose from C 1-4 Alkyl, C 3-6 Cycloalkyl, R 4 -(H2C) m- and R 5 -(H2C) n - the group formed;

[0095] in

[0096] m is 1;

[0097] n is 2;

[0098] R 4 C 3-6 Cycloalkyl;

[0099] R 5 For-OC 1-4 alkyl;

[0100] And the substituents A and R 2 As defined in any of the preceding embodiments.

[0101] Another embodiment of the present invention relates to a compound of formula (I),

[0102] in

[0103] R 1 Choose from C 1-4 Alkyl, C 3-4 Cycloalkyl, R 4 -(H2C) m - and R 5 -(H2C) n - the group formed;

[0104] in

[0105] m is 1;

[0106] n is 2;

[0107] R 4 C 3-4 Cycloalkyl;

[0108] R 5 For-OC 1-4 alkyl;

[0109] And the substituents A and R 2 As defined in any of the preceding embodiments.

[0110] Another embodiment of the present invention relates to a compound of formula (I),

[0111] in

[0112] R 1 Choose from C 1-4 Alkyl, C 3-4 Cycloalkyl, R 4 -(H2C) m - and R5 -(H2C) n - the group formed;

[0113] in

[0114] m is 1;

[0115] n is 2;

[0116] R 4 C 3-4 Cycloalkyl;

[0117] R 5 is H3C-O-;

[0118] And the substituents A and R 2 As defined in any of the preceding embodiments.

[0119] Another embodiment of the present invention relates to a compound of formula (I), wherein

[0120] R 1 Selected from the group consisting of: H3C, H3CH2C, H3COH2CH2C,

[0121] and

[0122] And the substituents A and R 2 As defined in any of the preceding embodiments.

[0123] Another embodiment of the present invention relates to a compound of formula (I), wherein

[0124] R 1 is H3C;

[0125] And the substituents A and R 2 As defined in any of the preceding embodiments.

[0126] Another embodiment of the present invention relates to a compound of formula (I),

[0127] in

[0128] R 2 Choose from H, C 1-4 Alkyl, C 3-6 Cycloalkyl, HO-C 1-4 Alkyl-, C 1-4 Fluoroalkyl, R 6 -(H2C) p - and R 7 -(H2C) q - the group formed;

[0129] in

[0130] p is 1;

[0131] q is 2;

[0132] R 6 Choose from C 3-6 the group consisting of cycloalkyl, C-morpholinyl, C-imidazolyl and C-pyrazolyl;

[0133] wherein the C-pyrazolyl, C-imidazolyl and C-morpholinyl are unsubstituted or C 1-4 Alkyl substitution;

[0134] R 7 Choose Free-OC 1-4 Alkyl, -OC 1-4 Fluoroalkyl, C 1-4 the group consisting of alkyl-S(O)2-, N-morpholinyl, N-imidazolyl and N-pyrazolyl;

[0135] wherein the N-pyrazolyl, N-imidazolyl, N-morpholinyl is unsubstituted or C 1-4 Alkyl substitution;

[0136] And the substituents A and R 1 As defined in any of the preceding embodiments.

[0137] Another embodiment of the present invention relates to a compound of formula (I), wherein

[0138] R 2 Choose from H, C 1-4 Alkyl, C 3-6 Cycloalkyl, HO-C 1-4 Alkyl-, C 1-2 Fluoroalkyl, R 6 -(H2C) p - and R 7 -(H2C) q - the group formed;

[0139] in

[0140] p is 1;

[0141] q is 2;

[0142] R 6 Choose from C 3-6 the group consisting of cycloalkyl, C-morpholinyl, C-imidazolyl and C-pyrazolyl;

[0143] wherein the C-pyrazolyl, C-imidazolyl and C-morpholinyl are unsubstituted or substituted with H3C;

[0144] R 7Selected from the group consisting of H3C-O-, -O-fluoromethyl, H3C-S(O)2-, N-morpholinyl, N-imidazolyl and N-pyrazolyl;

[0145] wherein the N-pyrazolyl, N-imidazolyl, and N-morpholinyl groups are unsubstituted or substituted with H3C;

[0146] And the substituents A and R 1 As defined in any of the preceding embodiments.

[0147] Another embodiment of the present invention relates to a compound of formula (I), wherein

[0148] R 2 Choose from H, C 1-4 Alkyl, C 3-6 Cycloalkyl, HO-C 1-4 Alkyl-, C 1-2 Fluoroalkyl, R 6 -(H2C) p - and R 7 -(H2C) q - the group formed;

[0149] in

[0150] p is 1;

[0151] q is 2;

[0152] R 6 Selected from the group consisting of: C 3-6 Cycloalkyl,

[0153] and

[0154]

[0155] R 7 Selected from the group consisting of: H3C-O, -O-fluoromethyl, H3C-S(O)2-,

[0156]

[0157] and

[0158]

[0159] And the substituents A and R 1 As defined in any of the preceding embodiments.

[0160] Another embodiment of the present invention relates to a compound of formula (I), wherein

[0161] R 2Selected from the group consisting of: H, H3C, H3CH2C, H3COH2CH2C, F2HCH2C, F3CH2C, FH2CH2C, H3C(O)2SH2CH2C, F3COH2CH2C,

[0162]

[0163] and

[0164]

[0165] And the substituents A and R 1 As defined in any of the preceding embodiments.

[0166] Another embodiment of the present invention relates to a compound of formula (I), wherein

[0167] R 2 is H;

[0168] And the substituents A and R 1 As defined in any of the preceding embodiments.

[0169] Preferred are compounds of formula (I) selected from the group consisting of:

[0170]

[0171]

[0172]

[0173]

[0174]

[0175] and

[0176] and substituent A is as defined in any one of the preceding embodiments.

[0177] Preferred are compounds according to formula (I) selected from

[0178]

[0179] and substituent A is as defined in any one of the preceding embodiments.

[0180] Especially preferred are compounds according to formula (I) selected from the group consisting of:

[0181]

[0182]

[0183]

[0184]

[0185]

[0186]

[0187]

[0188]

[0189]

[0190]

[0191]

[0192] and

[0193]

[0194] Terms and definitions used

[0195] Terms not specifically defined herein should be given the meanings given to them by those skilled in the art in view of the disclosure and context. However, unless otherwise specified, as used in this specification, the following terms have the designated meanings and will comply with the following conventions.

[0196] In the groups, radicals or moieties defined below, the number of carbon atoms is usually specified before the radical, e.g. "C 1-6 "Alkyl" means an alkyl group or alkyl radical having 1 to 6 carbon atoms. Generally speaking, in groups such as HO, H2N, (O)S, (O)2S, NC (cyano), HOOC, F3C or the like, one skilled in the art can see the point of radical attachment from the free valence of the group itself to the molecule. For composite groups containing two or more subunits, the last named subunit is the point of radical attachment, for example, the substituent "aryl-C 1-3 "Alkyl" means a C 1-3 Alkyl-bonded aryl, the C 1-3 The alkyl group is bonded to the nucleus or to the group to which the substituent is attached.

[0197] Where compounds of the invention are depicted in terms of both a chemical name and a chemical formula, in the event of any discrepancy the chemical formula will prevail. An asterisk may be used in a subformula to indicate a bond to the core molecule as defined.

[0198] The numbering of substituent atoms begins with the atom closest to the nucleus or closest to the group to which the substituent is attached.

[0199] For example, the term "3-carboxypropyl" represents the following substituent:

[0200]

[0201] wherein the carboxyl group is attached to the third carbon atom of the propyl group. The term "1-methylpropyl-", "2,2-dimethylpropyl-" or "cyclopropylmethyl-" refers to the following groups:

[0202]

[0203] Asterisks may be used in subformulas to indicate bonds to the core molecule as defined.

[0204] The term "C 1-n "alkyl" (wherein n is selected from an integer of 2, 3, 4 or 5) alone or in combination with another group means a non-cyclic saturated branched or straight chain hydrocarbon group having 1 to n C atoms. For example, the term "C 1-5 “Alkyl” encompasses the groups 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)-, H3C—CH2-CH(CH3)-, H3C—CH2-CH(CH3)-CH2—, H3C—CH(CH3)-CH2—, H3C—CH2-C(CH3)2—, H3C—C(CH3)2-CH2-CH2—, H3C—CH(CH3)-CH(CH3)-, and H3C—CH2-CH(CH2CH3)—.

[0205] The term "fluorine" added to an "alkyl," "alkylene," or "cycloalkyl" (saturated or unsaturated) group means such an alkyl or cycloalkyl group in which one or more hydrogen atoms are replaced by a fluorine atom. Examples include, but are not limited to, H2FC-, HF2C-, and F3C-.

[0206] The term "C 3-n "cycloalkyl" (wherein n is an integer from 4 to n), alone or in combination with another group, represents a cyclic, saturated, non-branched hydrocarbon group having 3 to n C atoms. For example, the term "C 3-6"Cycloalkyl" includes cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

[0207] The term halogen generally refers to fluorine, chlorine, bromine and iodine.

[0208] The term "phenyl" refers to a radical of the following ring:

[0209]

[0210] The term "thienyl" refers to a radical of the following ring:

[0211]

[0212] The term "benzothienyl" refers to a radical of the following ring:

[0213]

[0214] The term "benzofuranyl" refers to a radical of the following ring:

[0215]

[0216] The term "tetrazolyl" refers to a group of the following ring:

[0217]

[0218] The term "dioxodihydropyrimidinecarboxamide" refers to a radical of the following ring:

[0219]

[0220] The term "C-morpholinyl" refers to a group of the following ring:

[0221]

[0222] The term "C-imidazolyl" refers to a group of the following ring:

[0223]

[0224] The term "C-pyrazolyl" refers to a radical of the following ring:

[0225]

[0226] The term "N-morpholinyl" refers to a radical of the following ring:

[0227]

[0228] The term "N-imidazolyl" refers to a radical of the following ring:

[0229]

[0230] The term "N-pyrazolyl" refers to a radical of the following ring:

[0231]

[0232] As used herein, the term "substituted" means that any one or more hydrogens on the designated atom are replaced with a selected group from the designated group, provided that the designated atom's normal valence is not exceeded and that the substitution results in a stable compound.

[0233] Unless otherwise indicated, throughout the specification and the appended claims, a given chemical formula or name shall encompass tautomers and all stereoisomers, optical isomers, and geometric isomers (e.g., enantiomers, diastereomers, E / Z isomers, etc.), and racemates thereof, as well as mixtures of individual enantiomers in varying proportions, mixtures of diastereomers, or mixtures in which any of the foregoing forms of such isomers and enantiomers are present, and salts, including pharmaceutically acceptable salts thereof, and solvates thereof (such as hydrates including solvates of the free compound or solvates of the salt of the compound).

[0234] 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 using stereochemically pure starting materials and / or by stereoselective synthesis. It is known in the art how to prepare optically active forms, such as by resolution of racemic forms or by synthesis, for example starting with optically active starting materials and / or by using chiral reagents.

[0235] The enantiomerically pure compounds or intermediates of the present invention can be prepared via asymmetric synthesis, for example, by preparing and subsequently isolating appropriate diastereomeric compounds or intermediates which can be separated by known methods (for example, by chromatography or crystallization) and / or by using chiral reagents (such as chiral starting materials, chiral catalysts or chiral auxiliary agents).

[0236] Furthermore, the person skilled in the art knows how to prepare enantiomerically pure compounds from the corresponding racemic mixtures, such as by chromatographic separation of the corresponding racemic mixtures on a chiral stationary phase; or by resolution of the racemic mixtures using suitable resolving agents, for example, by means of formation of diastereomeric salts of the racemic compound with an optically active acid or base, followed by resolution of the salts and liberation of the desired compound from the salts; or by derivatization of the corresponding racemic compound with an optically active chiral auxiliary reagent, followed by separation of the diastereoisomers and removal of the chiral auxiliary group; or by kinetic resolution of the racemates (for example by enzymatic resolution); by enantiomer-selective crystallization from aggregates of isotropic crystals under appropriate conditions; or by (fractionated) crystallization from a suitable solvent in the presence of a photoactive chiral auxiliary.

[0237] The phrase "pharmaceutically acceptable" is used herein to refer to those compounds, substances, compositions, and / or dosage forms that are suitable for use within the scope of sound medical judgment without excessive toxicity, irritation, allergic response, or other problem or complication, and commensurate with a reasonable benefit / risk ratio.

[0238] As used herein, "pharmaceutically acceptable salts" refers to derivatives of the disclosed compounds wherein the parent compound forms a salt or complex with an acid or base. Examples of acids that form pharmaceutically acceptable salts with the parent compound containing a basic moiety include inorganic 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.

[0239] Examples of cations and bases that form pharmaceutically acceptable salts with the parent compound containing an acidic moiety include Na + , K + , Ca 2+ Mg 2+ NH4 + , L-arginine, 2,2'-iminobisethanol, L-lysine, N-methyl-D-glucosamine or phenyl(hydroxymethyl)-aminomethane. The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing a basic or acidic moiety by conventional chemical methods. Generally speaking, such salts can be prepared by reacting the free acid or free base form of these compounds with a sufficient amount of an aqueous solution or organic diluent solution (such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile) of the appropriate base or acid, or a mixture thereof.

[0240] In addition to those salts mentioned above, salts of other acids which are suitable, for example, for the purification or isolation of the compounds of the invention, such as trifluoroacetate, also form part of the present invention.

[0241] Biological analysis

[0242] Assessment of TRPA1 activity

[0243] Analysis A: TRPA1 analysis

[0244] The following in vitro TRPAl cell assay can be used to demonstrate the activity of compounds of the invention:

[0245] method:

[0246] Human HEK293 cells (Perkin Elmer, product number AX-004-PCL) overexpressing the human TRPA1 ion channel were used as a test system for compound efficacy and potency. Compound activity was determined by measuring the effect of the compound on intracellular calcium concentration induced by allyl isothiocyanate (AITC) agonism using a FLIPRtetra system (Molecular Devices).

[0247] Cell culture:

[0248] Cells were obtained as frozen cells in cryovials and stored at -150°C until use.

[0249] Cells were grown in culture medium (MEM / EBSS with 10% FCS and 0.4 mg / mL Geneticin). It was important that the density did not exceed 90% confluence. For subculture, cells were detached from the flask using Versene. One day before analysis, cells were detached, washed twice with culture medium (MEM / EBSS with 10% FCS), and 20,000 cells were seeded at 20 μl / well into 384-well poly-D-lysine biocoated plates from Corning (black with clear bottom, Cat. 356697). The plates were incubated at 37°C / 5% CO2 for 24 hours before use in the analysis.

[0250] Compound preparation

[0251] The test compound was dissolved in 100% DMSO at a concentration of 10 mM and first diluted in DMSO to a concentration of 5 mM, followed by serial dilution steps in 100% DMSO. The dilution factor and the number of dilution steps can be varied as needed. Eight different concentrations were typically prepared by a 1:5 dilution, with further intermediate dilutions (1:20) performed in HBSS / HEPES buffer (1× HEPES, Cat. 14065, Gibco; 20 mM HEPES, Cat. 83264, SIGMA; 0.1% BSA Cat. 11926, Invitrogen, pH 7.4).

[0252] FLIPR assay:

[0253] On the day of analysis, cells were washed 3 times with assay buffer, leaving 20 μL buffer in the wells after washing. 10 μL Ca6 kit (Cat.R8191, Molecular Devices) loading buffer in HBSS / HEPES was added to the cells and each dish was covered and incubated for 120 minutes at 37°C / 5% CO2. 10 μL compounds or controls in HBSS / HEPES buffer / 5% DMSO were carefully added to each well from the middle dilution dish. Luminescence (indicating calcium influx or release) was read on the FLIPRtetra device for 10 minutes to monitor the effects (e.g., agonism) induced by the compound. Finally, 10 μL of agonist AITC 50 μM (final concentration 10 μM) dissolved in HBSS / HEPES buffer / 0.05% DMSO was added to each well and then read for 10 minutes on the FLIPRtetra device. The area under the signal curve (AUC) after adding AITC was calculated for IC50 / inhibition %.

[0254] Data evaluation and calculation:

[0255] Each assay microtiter plate contained wells with vehicle (1% DMSO) control instead of compound as a control for AITC-induced luminescence (100% CTL; high control), and wells with vehicle control without AITC as a control for nonspecific changes in luminescence (0% CTL; low control).

[0256] Data analysis was performed by calculating the area under the signal curve for each well. Based on this value, the percentage of the measured value for each substance concentration was calculated using MegaLab software (developed in-house) (AUC(sample)-AUC(low))*100 / (AUC(high)-AUC(low)). The IC50 value was 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 = IC50M; b = Hill slope (hill); y = control %

[0257] Table 1: Biological data of compounds of the invention as obtained in Assay A

[0258]

[0259]

[0260] Table 2: Biological data of prior art compounds (Examples 28 and 29 in WO2017 / 060488) as obtained in Assay A.

[0261]

[0262] Table 3: Biological data of prior art compounds as obtained in assay A (Example 31 in L. Schenkel et al., J. Med. Chem. 2016, 59, 2794-2809).

[0263]

[0264] Assessment of microsomal clearance

[0265] Analysis B: Microsomal Clearance:

[0266] The metabolic degradation of the test compounds was analyzed using pooled liver microsomes at 37° C. The final incubation volume of 100 μl for each time point contained TRIS buffer at pH 7.6 (0.1 M) at RT, magnesium chloride (5 mM), microsomal protein (1 mg / ml), and the test compound at a final concentration of 1 μM.

[0267] After a short pre-incubation period at 37°C, the reaction was started by adding β-nicotinamide adenine dinucleotide phosphate in reduced form (NADPH, 1 mM) and terminated by transferring aliquots to a solvent at different time points (0, 5, 15, 30, 60 min). In addition, NADPH-independent degradation was monitored in cultures without NADPH and terminated at the last time point. The remaining test compound [%] after NADPH-independent cultivation was reflected by parameter c (control) (metabolic stability). The quenched cultures were assembled by centrifugation (10000 g, 5 min).

[0268] Aliquots of the supernatant were analyzed for parent compound by LC-MS / MS. The half-life (t1 / 2INVITRO) was determined by the slope of the semi-logarithmic plot of the concentration-time profile.

[0269] The intrinsic clearance (CL_INTRINSIC) was calculated by taking into account the amount of protein in the culture:

[0270] CL_INTRINSIC [μl / min / mg protein] = (Ln 2 / (half-life [min] * protein content [mg / ml])) * 1000

[0271] CL_INTRINSIC_INVIVO [ml / min / kg] = (CL_INTRINSIC [μl / min / mg protein] × MPPGL [mg protein / g liver] × hepatic factor [g / kg body weight]) / 1000

[0272] Qh[%]=CL[ml / min / kg] / liver blood flow[ml / min / kg])

[0273] Hepatocyte count, human: 120 × 10e6 cells / gram liver

[0274] Hepatic factor, human: 25.7 g / kg body weight

[0275] Blood flow, human: 21ml / (min×kg)

[0276] Table 4: Biological data of compounds of the invention obtained in Assay B

[0277]

[0278]

[0279] Table 5: Biological data of prior art compounds (Examples 28 and 29 in WO2017 / 060488) as obtained in Assay B.

[0280]

[0281]

[0282] Table 6: Biological data of prior art compounds (Example 31 in L. Schenkel et al., J. Med. Chem. 2016, 59, 2794-2809) as obtained in Assay B.

[0283]

[0284] Assessment of hepatocyte clearance

[0285] Analysis C: Hepatocellular Clearance

[0286] Metabolic degradation of test compounds was analyzed in hepatocyte suspensions. Hepatocytes (cryopreserved) were cultured in Dulbecco's modified eagle medium (supplemented with 3.5 μg glucagon / 500 mL, 2.5 mg insulin / 500 mL, and 3.75 mg / 500 mL hydrocortisone) containing 5% or 50% species serum.

[0287] After pre-incubation for 30 min in an incubator (37°C, 10% CO2), 5 μl of test compound solution (80 μM; diluted 1:25 from a 2 mM DMSO stock solution with culture medium) was added to 395 μl of hepatocyte suspension (cell density ranging from 0.25 to 5 million cells / mL, depending on the species, typically 1 million cells / mL; final concentration of test compound was 1 μM, final DMSO concentration was 0.05%).

[0288] The cells were incubated for six hours (incubator, orbital shaker) and samples (25 μl) were taken at 0, 0.5, 1, 2, 4, and 6 hours. The samples were transferred to acetonitrile and pelleted by centrifugation (5 min). The supernatant was transferred to a new 96-deep-well plate, evaporated under nitrogen, and resuspended.

[0289] Reduction of parent compound analyzed by HPLC-MS / MS

[0290] CLint is calculated as follows: CL_INTRINSIC = dose / AUC = (C0 / CD) / (AUD+clast / k)×1000 / 60. C0: initial concentration in culture [μM], CD: cell density of viable 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 for parent reduction [h-1].

[0291] The calculated in vitro hepatic intrinsic clearance can be scaled up to the in vivo hepatic intrinsic clearance and used to predict the in vivo hepatic blood clearance (CL) by using a liver model (a well-stirred model).

[0292] CL_INTRINSIC_INVIVO [ml / min / kg] = (CL_INTRINSIC [μL / min / 10e6 cells] × number of hepatocytes [10e6 cells / gram of liver] × hepatic factor [g / kg body weight]) / 1000

[0293] CL [ml / min / kg] = CL_INTRINSIC_INVIVO [ml / min / kg] × liver blood flow [ml / min / kg] / (CL_INTRINSIC_INVIVO [ml / min / kg] + liver blood flow [ml / min / kg])

[0294] Qh[%]=CL[ml / min / kg] / liver blood flow[ml / min / kg])

[0295] Hepatocyte count, human: 120 × 10e6 cells / gram liver

[0296] Hepatic factor, human: 25.7 g / kg body weight

[0297] Blood flow, human: 21ml / (min×kg)

[0298] Table 7: Biological data of compounds of the invention as obtained in Assay C

[0299]

[0300]

[0301] Table 8: Biological data of prior art compounds (Examples 28 and 29 in WO2017 / 060488) as obtained in Assay C.

[0302]

[0303] Table 9: Biological data of prior art compounds as obtained in assay C (Example 31 in L. Schenkel et al., J. Med. Chem. 2016, 59, 2794-2809).

[0304]

[0305] Permeability assessment

[0306] Caco-2 cells (1-2×105 cells / 1cm 2 area) were seeded on filter inserts (Costar transwell polycarbonate or PET filters, 0.4 μm pore size) and cultured (DMEM) for 10 to 25 days.

[0307] Compounds were dissolved in an appropriate solvent (e.g., DMSO, 1-20 mM stock solution). The stock solution was 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 solutions (0.1-300 μM compound, final DMSO <= 0.5%). The transport solution (TL) was applied to the apical or basolateral donor side for measurement of AB or BA permeability, respectively (filtration was repeated three times). Samples were collected from the donor at the beginning and end of the experiment and from the acceptor side at various time intervals for 2 hours for concentration measurement by HPLC-MS / MS or scintillation counting. Replace the sampled receptor volume with fresh receptor solution.

[0308] Assessment of plasma protein binding

[0309] This equilibrium dialysis (ED) technique is used to measure the approximate in vitro fractionation binding of test compounds to plasma proteins. Use Dianorm Teflon dialysis unit (0.2 ml). Each unit consists of a donor chamber and a receptor chamber, separated by an ultrathin semipermeable membrane with a 5 kDa molecular weight cutoff. A stock solution for each test compound is prepared with 1 mM in DMSO and diluted to a final concentration of 1.0 μM. Subsequent dialysis solution is prepared in human or rat plasma (containing NaEDTA) from the merger of male and magnetic donors. An aliquot of 200 μL dialysis buffer (100 mM potassium phosphate, pH 7.4) is distributed in the buffer chamber. An aliquot of 200 μL test compound dialysis solution is distributed in the plasma chamber. Cultivate 2 hours at 37 ° C under rotation.

[0310] 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 of ACN / water (80 / 20). A 25 μL aliquot of plasma dialysate was transferred to a deep well plate and mixed with 25 μL of ACN / water (80 / 20), 25 μL of buffer, 25 μL of calibration solution, and 25 μL of internal standard solution. Protein precipitation was performed by adding 200 μL of ACN. A 50 μL aliquot of buffer dialysate was transferred to a deep well plate and mixed with 25 μL of blank plasma, 25 μL of internal standard solution, and 200 μL of ACN. Samples were measured on an HPLC-MS / MS-system and evaluated with the aid of Analyst-Software. The percent binding was calculated using the following formula: % binding = (plasma concentration - buffer concentration / plasma concentration) × 100.

[0311] Solubility assessment

[0312] A saturated solution is prepared in a well plate (format depends on the robot) by adding an appropriate volume of the selected liquid medium (usually in the range of 0.25-1.5 ml) to each well containing a known amount of solid API (usually in the range of 0.5-5.0 mg). Each well is shaken or stirred for a predetermined period of time (usually in the range of 2-24 h) and then filtered using an appropriate filter membrane (usually a PTFE filter membrane with a pore size of 0.45 μm). Absorption of the filter membrane is avoided by discarding the first few drops of filtrate. The amount of dissolved API is determined by UV spectroscopy. In addition, a glass electrode pH meter is used to measure the pH of the saturated aqueous solution.

[0313] Evaluation of Pharmacokinetic Characteristics in Rodents

[0314] Test compounds were administered intravenously to fed rats or orally to fasted rats. Blood samples were obtained at several time points after administration of the test compounds, anticoagulated, and centrifuged.

[0315] The concentration of the administered compound and / or metabolite was quantified as the analyte in plasma samples. PK parameters were calculated using non-compartmental methods. AUC and Cmax were normalized to a dose of 1 μmol / kg.

[0316] Assessment of human hepatocyte metabolism in vitro

[0317] Primary human hepatocytes in suspension were used to investigate the metabolic pathways of test compounds. After recovery from cryopreservation, human hepatocytes were cultured 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.

[0318] After pre-incubation for 30 min in a cell culture incubator (37° C., 10% CO 2 ), the test compound solution was incorporated into the hepatocyte suspension to obtain a 1.0*10 6 Up to 4.0*10 6 The cells were then cultured at a final cell density of 10 cells / mL (depending on the metabolic turnover rate of the compound observed with primary human hepatocytes), a final test compound concentration of 10 μM, and a final DMSO concentration of 0.05%.

[0319] Cells were incubated for six hours in a cell culture incubator on a horizontal shaker, and samples were removed from the incubation after 0, 0.5, 1, 2, 4, or 6 hours, depending on metabolic turnover. Samples were quenched with acetonitrile and pelleted by centrifugation. The supernatant was transferred to a 96-deep-well plate, evaporated under nitrogen, and resuspended before bioanalysis by liquid chromatography-high-resolution mass spectrometry for identification of putative metabolites.

[0320] Based on Fourier-Transform-MS n The data are tentatively assigned structures. Metabolites are reported as a percentage of the parent in human hepatocyte culture, with a cutoff of ≥4%.

[0321] Treatment

[0322] The present invention relates to compounds of formula 1, which are suitable for preventing and / or treating diseases and / or conditions associated with or regulated by TRPA1 activity, including but not limited to treating and / or preventing fibrotic diseases, inflammatory and immunomodulatory disorders, respiratory or gastrointestinal diseases or discomfort, ophthalmic diseases, inflammatory diseases of the joints and inflammatory diseases of the nasopharynx, eyes and skin, and pain and neurological disorders. Such conditions, diseases and discomforts include cough, idiopathic pulmonary fibrosis, other interstitial lung diseases and other fibrotic changes, asthma or allergic diseases, eosinophilic diseases, chronic obstructive pulmonary disease, and inflammatory and immunomodulatory disorders (such as rheumatoid arthritis and atherosclerosis), as well as pain and neurological disorders (such as acute pain, surgical pain, chronic pain and depression, and bladder disorders).

[0323] The compounds of formula 1 are suitable for the prevention and / or treatment of:

[0324] (1) Cough, such as chronic idiopathic cough or chronic refractory cough, cough associated with asthma, COPD, lung cancer, post-viral infection, idiopathic pulmonary fibrosis and other interstitial lung diseases.

[0325] (2) Pulmonary fibrosis (such as pneumonia or interstitial pneumonia associated with collagen disease), for example, lupus erythematosus, systemic scleroderma, rheumatoid arthritis, polymyositis and dermatomyositis, idiopathic interstitial pneumonia (such as idiopathic pulmonary fibrosis (IPF)), nonspecific interstitial pneumonia, interstitial lung disease associated with respiratory bronchiolitis, desquamative interstitial pneumonia, cryptogenic organizing pneumonia, acute interstitial pneumonia and lymphocytic interstitial pneumonia, lymphangiomyomatosis, pulmonary alveolar proteinosis, Langerhan's cell histiocytosis, pleural parenchymal fibroelastosis, interstitial lung disease of known etiology, such as that due to occupational exposure (such as asbestosis, silicosis, miner's lung (coal dust), farmer's lung (hay and mold), pigeon fanciers' lung (piggeon fanciers' lung) lung) (birds) or other occupational airborne causes (such as metal dust or mycobacteria) or due to treatment (such as radiation, methotrexate, amiodarone, nitrofurantoin or chemotherapeutic agents) or interstitial pneumonia or granulomatous diseases such as granulomatosis with polyangiitis, Church-Strauss syndrome, sarcoidosis, hypersensitivity pneumonitis, or interstitial pneumonia caused by a different etiology (e.g., inhalation of toxic gases, vapors, bronchitis or pneumonia) or interstitial pneumonia caused by heart failure, X-rays, radiation, chemotherapy, M. boeck's disease or sarcoidosis, granulomatous diseases, cystic fibrosis or mucoid disease or alpha-1 antitrypsin deficiency.

[0326] (3) Other fibrotic diseases, such as bridging fibrosis of the liver, cirrhosis, nonalcoholic steatohepatitis (NASH), atrial fibrosis, endomyocardial fibrosis, old myocardial infarction, glial scar, arthrofibrosis, Dupuytren's contracture, keloid, scleroderma / systemic sclerosis, mediastinal fibrosis, myelofibrosis, Peyronie's disease, nephrogenic systemic fibrosis, retroperitoneal fibrosis, and adhesive capsulitis.

[0327] (4) Inflammatory, autoimmune or allergic diseases and conditions, such as allergic or non-allergic rhinitis or sinusitis, chronic sinusitis or rhinitis, nasal polyps, chronic sinusitis, acute sinusitis, asthma, childhood asthma, allergic bronchitis, alveolitis, hyperreactive trachea, allergic conjunctivitis, bronchiectasis, adult respiratory distress syndrome, bronchial and pulmonary edema, bronchitis or localized 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), syndrome), delayed hypersensitivity, non-allergic asthma; exercise-induced bronchoconstriction; chronic obstructive pulmonary disease (COPD), acute bronchitis, chronic bronchitis, cough, emphysema; systemic anaphylaxis or hypersensitivity reactions, drug allergy (e.g., to penicillins, cephalosporins), eosinophilic myalgia syndrome due to ingestion of contaminated tryptophan, insect sting hypersensitivity; autoimmune diseases such as rheumatoid arthritis, Graves' disease, Sjogren's syndrome, psoriatic arthritis, multiple sclerosis, systemic lupus erythematosus, myasthenia gravis, immune thrombocytopenia (adult ITP / neonatal ITP, pediatric ITP), immune hemolytic anemia (autoimmune and drug-induced), Evans syndrome (immune thrombocytopenia of platelets and red blood cells), neonatal Rhinopathies, Goodpasture's syndrome syndrome (anti-GBM disease), celiac disease, autoimmune cardiomyopathy, juvenile-onset diabetes mellitus; glomerulonephritis, autoimmune thyroiditis, Behcet's disease; transplant rejection (e.g., in transplantation), including allograft rejection or graft-versus-host disease; inflammatory bowel diseases, such as Crohn's disease and ulcerative colitis; spondyloarthropathies; psoriasis (including T-cell-mediated psoriasis) and inflammatory skin diseases (such as dermatitis, eczema, atopic dermatitis, allergic contact dermatitis, urticaria); vasculitis (e.g., necrotizing, cutaneous, and hypersensitivity vasculitis); erythema nodosum; eosinophilic myositis, eosinophilic fasciitis, and cancers of the skin or organs with leukocytic infiltration;Ophthalmological diseases such as age-related macular degeneration, diabetic retinopathy and diabetic macular edema, keratitis, eosinophilic keratitis, keratoconjunctivitis, vernal keratoconjunctivitis, scars, anterior segment scarring, blepharitis, blepharoconjunctivitis, bullous disorders, cicatricial pemphigoid, conjunctival melanoma, papillary conjunctivitis, dry eye, episcleritis, glaucoma, gliomatosis, annular granuloma, Graves' ophthalmopathy, intraocular melanoma, conjunctival macula, proliferative vitreoretinopathy, pterygium, scleritis, acute gout attack, gout, or osteoarthritis.

[0328] (5) Pain, such as chronic idiopathic pain syndrome, neuralgia, dysesthesia, allodynia, migraine, dental pain and postoperative pain.

[0329] (6) Depression, anxiety, diabetic neuropathy and bladder disorders such as bladder outlet obstruction, overactive bladder, cystitis; myocardial reperfusion injury or cerebral ischemic injury.

[0330] Therefore, the present invention relates to a compound of formula 1 for use as a medicament.

[0331] Furthermore, the present invention relates to the use of a compound of formula 1 for treating and / or preventing diseases and / or conditions associated with or regulated by TRPA1 activity.

[0332] Furthermore, the present invention relates to the use of compounds of formula 1 for the treatment and / or prevention of fibrotic diseases, inflammatory and immunomodulatory disorders, respiratory or gastrointestinal diseases or complaints, ophthalmic diseases, inflammatory diseases of the joints and inflammatory diseases of the nasopharynx, eyes and skin, pain and neurological disorders. Such disorders, diseases and complaints include cough, idiopathic pulmonary fibrosis, other interstitial lung diseases and other fibrotic conditions, asthma or allergic diseases, eosinophilic diseases, chronic obstructive pulmonary disease, and inflammatory and immunomodulatory disorders (such as rheumatoid arthritis and atherosclerosis), as well as pain and neurological disorders (such as acute pain, surgical pain, chronic pain and depression, and bladder disorders).

[0333] Furthermore, the present invention relates to the use of a compound of formula 1 for treating and / or preventing:

[0334] (1) Cough, such as chronic idiopathic cough or chronic refractory cough, cough associated with asthma, COPD, lung cancer, post-viral infection, idiopathic pulmonary fibrosis and other interstitial lung diseases.

[0335] (2) Pulmonary fibrosis (such as pneumonia or interstitial pneumonia associated with collagen disease), for example, lupus erythematosus, systemic scleroderma, rheumatoid arthritis, polymyositis and dermatomyositis, idiopathic interstitial pneumonia (such as idiopathic pulmonary fibrosis (IPF)), nonspecific interstitial pneumonia, interstitial lung disease associated with respiratory bronchiolitis, desquamative interstitial pneumonia, cryptogenic organizing pneumonia, acute interstitial pneumonia and lymphocytic interstitial pneumonia, lymphangiomyomatosis, pulmonary alveolar proteinosis, Langerhans cell histiocytosis, pleural parenchymal fibroelastosis, interstitial lung disease of known etiology, such as due to occupational exposure (such as asbestosis, silicosis, miners' Interstitial pneumonia or granulomatous diseases such as granulomatosis with polyangiitis, Church-Strauss syndrome, sarcoidosis, hypersensitivity pneumonitis, or interstitial pneumonia caused by a different etiology (e.g., inhalation of toxic gases, vapors, bronchitis, or pneumonia), or caused by heart failure, X-rays, radiation, chemotherapy, Burke's disease or sarcoidosis, granulomatous diseases, cystic fibrosis or mucoid disease, or alpha-1 antitrypsin deficiency.

[0336] (3) Other fibrotic diseases, such as bridging fibrosis of the liver, cirrhosis, nonalcoholic steatohepatitis (NASH), atrial fibrosis, endomyocardial fibrosis, old myocardial infarction, glial scar, arterial stiffness, arthrofibrosis, Dupuytren's contracture, keloid, scleroderma / systemic sclerosis, mediastinal fibrosis, myelofibrosis, Peyronie's disease, nephrogenic systemic fibrosis, retroperitoneal fibrosis, and adhesive capsulitis.

[0337] (4) Inflammatory, autoimmune or allergic diseases and conditions, such as allergic or non-allergic rhinitis or sinusitis, chronic sinusitis or rhinitis, nasal polyps, chronic sinusitis, acute sinusitis, asthma, childhood asthma, allergic bronchitis, alveolitis, hyperreactive trachea, allergic conjunctivitis, bronchiectasis, adult respiratory distress syndrome, bronchial and pulmonary edema, bronchitis or localized pneumonia, eosinophilic cellulitis (e.g., Wells syndrome), eosinophilic pneumonia (e.g., Loeffler syndrome, chronic eosinophilic pneumonia), eosinophilic fasciitis (e.g., Scheuermann syndrome), delayed hypersensitivity, non-allergic asthma; exercise-induced bronchoconstriction atrophy; chronic obstructive pulmonary disease (COPD), acute bronchitis, chronic bronchitis, cough, emphysema; systemic allergic reaction or hypersensitivity reaction, drug allergy (e.g., to penicillins, cephalosporins), eosinophilic myalgia syndrome caused by ingestion of contaminated tryptophan, insect sting allergy; autoimmune diseases such as rheumatoid arthritis, Graves' disease, Sjögren's syndrome, psoriatic arthritis, multiple sclerosis, systemic lupus erythematosus, myasthenia gravis, immune thrombocytopenia (adult ITP / neonatal thrombocytopenia, pediatric ITP), immune hemolytic anemia (autoimmune and drug-induced), Ivan syndrome (immune thrombocytopenia of platelets and red blood cells), neonatal Rh disease, Cuban syndrome (anti-GBM disease), celiac disease, autoimmune cardiomyopathy, juvenile-onset diabetes mellitus; glomerulonephritis, autoimmune thyroiditis, Behçet's disease; transplant rejection (e.g., in transplantation), including allograft rejection or graft-versus-host disease; inflammatory bowel diseases, such as Crohn's disease and ulcerative colitis; spondyloarthropathies; psoriasis (including T-cell-mediated psoriasis) and inflammatory skin diseases (such as dermatitis, eczema, atopic dermatitis, allergic contact dermatitis, urticaria); vasculitis (e.g., necrotizing, cutaneous, and hypersensitivity vasculitis) ); erythema nodosum; eosinophilic myositis, eosinophilic fasciitis, cancer of the skin or organs with leukocyte infiltration; ophthalmological diseases such as age-related macular degeneration, diabetic retinopathy and diabetic macular edema, keratitis, eosinophilic keratitis, keratoconjunctivitis, vernal keratoconjunctivitis, scars, anterior segment scars, blepharitis, blepharoconjunctivitis, bullous disorders, cicatricial pemphigoid, conjunctival melanoma, papillary conjunctivitis, dry eye, episcleritis, glaucoma, gliomatosis, annular granuloma, Grave's ophthalmopathy, intraocular melanoma, conjunctival macula, proliferative vitreoretinopathy, pterygium, scleritis, acute gout attack, gout, or osteoarthritis.

[0338] (5) Pain, such as chronic idiopathic pain syndrome, neuralgia, dysesthesia, allodynia, migraine, dental pain and postoperative pain.

[0339] (6) Depression, anxiety, diabetic neuropathy and bladder disorders such as bladder outlet obstruction, overactive bladder, cystitis; myocardial reperfusion injury or cerebral ischemic injury.

[0340] In another aspect, the present invention relates to a compound of formula 1 for use in the treatment and / or prevention of the diseases and conditions mentioned above.

[0341] In another aspect, the present invention relates to the use of a compound of formula 1 for the preparation of a medicament for the treatment and / or prevention of the diseases and conditions mentioned above.

[0342] In another aspect of the present invention, the present invention relates to a method for treating or preventing the above-mentioned diseases and conditions, which comprises administering an effective amount of a compound of formula 1 to a human.

[0343] Combination therapy

[0344] 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 consisting of therapeutic agents suitable for treating the diseases or conditions described above, in particular those associated with fibrotic diseases, inflammatory and immunomodulatory disorders, diseases or complaints of the respiratory or gastrointestinal tract, 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 autoimmune disorders (such as rheumatoid arthritis and atherosclerosis); or therapeutic agents suitable for treating ophthalmic diseases, pain and depression.

[0345] Suitable additional therapeutic agents for such combinations include in particular those which, for example, enhance the therapeutic effect of one or more active substances for one of the mentioned indications and / or allow a dosage reduction of one or more active substances.

[0346] Thus, the compounds of the present invention may be combined with one or more additional therapeutic agents selected from the group consisting of an antifibrotic agent, an antitussive agent, an anti-inflammatory agent, an anti-atopic dermatitis agent, an analgesic, an anticonvulsant, an anxiolytic, a sedative, a skeletal muscle relaxant, or an antidepressant.

[0347] Antifibrotic agents are, for example, nintedanib, pirfenidone, phosphodiesterase type 4 inhibitors (PDE4) (such as roflumilast), autotaxin inhibitors such as GLPG-1690 or BBT-877; connective tissue growth factor (CTGF) blocking antibodies such as pamrevlumab; B-cell activating factor receptor (BAFF-R) blocking antibodies such as lanalumab; alpha-v / beta-6 blocking inhibitors (such as BG-00011 / STX-100), recombinant pentraxin-2 (PTX-2) (such as PRM-151); c-Jun JNK inhibitors, such as CC-90001; 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 protein kinase 2 (ROCK2), such as KD-025; heat shock protein 47 (HSP47) small interfering RNA / ND-L02-s0201, such as BMS-986263; Wnt pathway inhibitors, such as SM-04646; LD4 / PDE3 / 4 inhibitors, such as Tipelukast; recombinant immunomodulatory domains of histidyl tRNA synthetase (HARS), such as ATYR-1923; prostaglandin synthase inhibitors, such as ZL-2102 / SAR-191 801; 15-hydroxy-eicosapentaenoic acid (15-HEPE, e.g., DS-102); lysyl oxidase-like 2 (LOXL2) inhibitors, such as PAT-1251, PXS-5382 / PXS-5338; phosphatidylinositol 3-kinase (PI3K) / mammalian target of rapamycin (mTOR) dual inhibitors, such as HEC-68498; calpain inhibitors, such as BLD-2660; mitogen-activated protein 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 β1 (TGF-β1) small interfering RNA, such as TRK250 / BNC-1021; or lysophosphatidic acid receptor antagonists, such as BMS-986278.

[0348] For example, the antitussive agent is a purinergic receptor 3 (P2X3) receptor antagonist, such as gefapixant, S-600918, BAY-1817080 or BLU-5937; a neurokinin 1 (NK-1) receptor antagonist, such as orvepitant, aprepitant; a nicotinic acetylcholine receptor α7 subunit stimulator, such as ATA-101 / bradanicline, codeine, gabapentin, pregablin or azithromycin. For example, the anti-inflammatory agent is a corticosteroid such as prednisolone or dexamethasone; a cyclooxygenase-2 (COX2) inhibitor such as celecoxib, rofecoxib, parecoxib, valdecoxib, deracoxib, etoricoxib or lumiracoxib; prostaglandin E2 antagonists; leukotriene B4 antagonists; leukotriene D4 antagonists, such as monteleukast; 5-lipoxygenase inhibitors; or other nonsteroidal anti-inflammatory agents (NSAIDs), such as aspirin, diclofenac, diflunisal, etodolac, ibuprofen, or indomethacin.

[0349] For example, anti-atopic dermatitis agents include cyclosporin, 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), IL-13 (e.g., lebrikizumab, tralokinumab), and IL-31 (nemolizumab).

[0350] For example, the analgesic is an opioid such as morphine base, oxymorphine, levopanol, oxycodone, propoxyphene, nalmefene, fentanyl, hydrocodone, hydromorphone, meripidine, methadone, nalorphine, naloxone, naltrexone, buprenorphine, butorphanol, nalbuphine, pentazocine; or a non-opioid such as acetylcholine.

[0351] For example, the antidepressant is a tricyclic antidepressant such as amitriptyline, clomipramine, despramine, doxepin, desipramine, imipramine, nortriptyline; a selective serotonin reuptake inhibitor antidepressant (SSRI) such as fluoxetine, paroxetine, sertraline, citalopram, escitalopram; a norepinephrine reuptake inhibitor antidepressant (SNRI) such as maprotiline, lofepramine, mirtazapine, oxaprotiline, fezolamine, tomoxetine; antidepressants (such as fentanyl, fentanyl, bromocriptine ...

[0352] For example, the anxiolytic is a benzodiazepine such as alprazolam, bromazepam, chlordiazepoxide, clonazepam, clozapine, diazepam, flurazepam, lorazepam, oxazepam, temazepam, triazolam, or tofisopam; or it is a non-benzodiazepine. or a diazepine hypnotic such as eszopiclone, zaleplon, zolpidem or zopiclone; or a carbamate such as meprobamate, carisoprodol, tybamate or lorbamate; or an antihistamine such as hydroxyzine, chlorpheniramine or diphenhydramine.

[0353] For example, the sedative is a barbiturate sedative such as amobarbital, aprobarbital, butabarbital, butabital, mephobarbital, metharbital, methohexital, pentobarbital, secobarbital, talbutal, theamylal or thiopental; or a non-barbiturate sedative such as glutethimide, meprobamate, methaqualone or dichloalphenazone.

[0354] For example, the skeletal muscle relaxant is baclofen, meproba, carbothiazide, cyclobenzaprine, metaxalone, methocarbamol, tizanidine, chlorzoxazone or orphenadrine.

[0355] Other suitable combination partners are acetylcholinesterase inhibitors, such as donepezil; 5-HT-3 antagonists, such as ondansetron; metabotropic glutamate receptor antagonists; antiarrhythmics, such as mexiletine or phenytoin; or NMDA receptor antagonists.

[0356] Other suitable combination partners are incontinence medications, for example anticholinergics such as oxybutynin, tolterodine, darifenacin, fesoterodine, solifenacin or trospium; or bladder muscle relaxants such as mirabegron; or alpha blockers such as tamsulosin, alfuzosin, silodosin, doxazosin or terazosin.

[0357] The dosage of the above combination is usually 1 / 5 of the normal recommended minimum dose to 1 / 1 of the normal recommended dose.

[0358] Therefore, in a further aspect, the present invention relates to the use of a compound according to the invention in combination with one or more additional therapeutic agents as described above or below for the treatment of diseases or conditions that can be influenced or mediated by TRPAl, in particular diseases or conditions as described above or below.

[0359] In another aspect, the present invention relates to a method for treating a disease or condition in a patient that is amenable to TRPAl inhibition, 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 pharmaceutically acceptable salt thereof in combination with a therapeutically effective amount of one or more additional therapeutic agents.

[0360] 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 treating a disease or condition amenable to TRPAl inhibition in a patient in need thereof.

[0361] In a further aspect, the present invention relates to a method for treating 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 above and below.

[0362] The use of the compounds according to the invention in combination with other therapeutic agents may be carried out simultaneously or at staggered times.

[0363] The compound according to the invention and the additional therapeutic agent(s) may both be present together in one formulation (e.g. a tablet or capsule) or separately in two identical or different formulations (e.g. in a so-called kit-of-parts).

[0364] Thus, in another aspect, the present invention relates to a pharmaceutical composition comprising a compound according to the invention and one or more additional therapeutic agents as described above or below, optionally together with one or more inert carriers and / or diluents.

[0365] In a further aspect, the invention relates to the use of a compound according to the invention in a cough measurement device.

[0366] Other features and advantages of the present invention will become apparent from the following more detailed examples, which illustrate by way of example the principles of the invention.

[0367] preparation

[0368] The compounds of the present invention and their intermediates can be obtained using synthetic methods known to those skilled in the art and described in the literature on organic synthesis. Preferably, the compounds are obtained in a manner similar to the preparation methods explained more fully below, especially as described in the experimental section. In some cases, the order in which the reaction steps are carried out may be varied. Variations of reaction methods known to those skilled in the art but not described in detail herein may also be used.

[0369] The general method for preparing the compounds according to the present invention will become apparent to those skilled in the art by studying the following schemes. Conventional protecting groups may be used to protect any functional group in the starting materials or intermediates. These protecting groups may be cleaved again at a suitable stage in the reaction sequence using methods familiar to those skilled in the art.

[0370] The compounds according to the present invention are prepared by the synthetic methods described below, wherein the substituents of the general formula have the meanings given above. These methods are intended to illustrate the present invention, but do not limit the scope of the compounds claimed in the subject matter and these examples. Where the preparation of the starting compound is not described, it is commercially available or can be prepared in a manner similar to the known compounds or methods described herein. The substances described in the literature are prepared according to the published synthetic methods. Abbreviations are as defined in the Examples section.

[0371] Process 1:

[0372]

[0373] In Scheme 1, chloromethyltetrazole is N-alkylated with an appropriate ethyl ketone derivative carrying a leaving group "LG" (e.g., Cl or Br) adjacent to the carbonyl group in the presence of a base (e.g., KCO) to provide a mixture of two regioisomers. The undesired regioisomer can be removed by chromatography using an appropriate gradient (not shown). The resulting ketone (A) can be reduced in an enantioselective manner to provide the alcohol (B) using an appropriate catalytic system using a transition metal complex (e.g., Ru or Ir) in combination with a chiral ligand (e.g., ([(1S,2S)-2-amino-1,2-diphenylethyl](4-tosyl)amide) and a hydrogen source (e.g., triethylamine formate complex).

[0374] Uracil derivatives (D) can be synthesized neat from monosubstituted ureas and 1,3-diethyl 2-(ethoxymethylene)malonate at elevated temperatures to directly produce (D), or to produce (C), which can be further reacted under alkaline conditions such as NaOEt in EtOH at elevated temperatures to yield (D). Primary amides (E) can be synthesized from esters (D) by stirring with ammonia in a solvent such as water or an alcohol at elevated temperatures in a sealed container.

[0375] The final compound (I) can be synthesized by alkylating (E) with intermediate (B) in the presence of a base such as K2CO3. Alternatively, intermediate (D) is alkylated with (B) in the presence of a base to give (F), which can be hydrolyzed with a suitable reagent such as LiOH to give acid (G). Acid (G) can then be coupled with an amine in the presence of an amide coupling agent such as HATU and in the presence of a base such as DIPEA to give the final compound (I).

[0376] Alternatively, compounds of formula (I) can be prepared as shown in Scheme 2 below.

[0377] Process 2:

[0378]

[0379] In Scheme 2, intermediate (H) can be prepared by alkylating (E) with an acetonitrile derivative carrying a leaving group "LG" (e.g., Cl or Br) in the presence of a base such as DIPEA. The formation of tetrazole (J) can be achieved by typical reaction conditions for tetrazole formation (e.g., using NaN3 in the presence of TEA / TEA hydrochloride in DMF). Alkylation of tetrazole (J) is carried out with an appropriate ethyl ketone derivative carrying a leaving group "LG" (e.g., Cl or Br) adjacent to the carbonyl group in the presence of a base such as DIPEA to provide a mixture of two regioisomers. Undesired regioisomers can be removed by chromatography using an appropriate gradient (not shown). Finally, the keto group of (K) can be reduced in an enantioselective manner by using a suitable catalytic system using a transition metal complex (e.g., Ru or Ir) in combination with a chiral ligand (e.g., ([(1S,2S)-2-amino-1,2-diphenylethyl](4-tosyl)amide) and a hydrogen source (e.g., triethylamine formate complex) to afford the final compound (I). Alternatively, the final compound (I) can be prepared by alkylating the intermediate (J) with a suitable aromatic or heteroaromatic alcohol derivative carrying a leaving group "LG" (e.g., Cl or Br) adjacent to the hydroxyl group in the presence of a base such as DIPEA, and subsequently isolating the desired regioisomer.

[0380] Example

[0381] preparation

[0382] Compounds according to the present invention and intermediates thereof can be prepared using synthetic methods known to those skilled in the art and described in the literature of organic synthesis, for example, using 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 compound is obtained by a preparation method similar to that described more fully below, particularly as described in the experimental section. In some cases, the order in which the reaction scheme is implemented may vary. Variations of these reactions known to those skilled in the art but not described in detail herein may also be used. Based on studying the following flow, the general method for preparing compounds according to the present invention will become apparent to those skilled in the art. The starting compound is commercially available or can be prepared by methods described in the literature or herein, or can be prepared in a similar or similar manner. Before reacting, any corresponding functional group in the starting compound can be protected using conventional protecting groups. These protecting groups can be cleaved again at an appropriate stage in the reaction sequence using methods familiar to those skilled in the art and described in, for example, "Protecting Groups", 3rd edition, Philip J. Kocienski, Thieme, 2005 and "Protective Groups in Organic Synthesis", 4th edition, Peter G.M. Wuts, Theodora W. Greene, John Wiley & Sons, 2006. The terms "ambient temperature" and "room temperature" are used interchangeably and refer to a temperature of about 20° C., for example, between 19 and 24° C.

[0383] abbreviation:

[0384]

[0385]

[0386] Preparation of intermediates

[0387] Intermediate I

[0388] Intermediate I.1 (General Procedure)

[0389] 2-[5-(Chloromethyl)-2H-1,2,3,4-tetrazol-2-yl]-1-(4-chlorophenyl)ethan-1-one

[0390]

[0391] At RT, 1.63 g (11.8 mmol) of K CO was added to a 15 mL DMA solution of 1.00 g (8.44 mmol) of 5-(chloromethyl)-2H-1,2,3,4-tetrazole and 2.17 g (9.28 mmol) of 4-chlorophenacyl bromide under stirring. The reaction mixture was stirred for 30 min at RT and then filtered. The filtrate was diluted with water and a saturated aqueous NaCl solution and extracted three times with EtOAc. The combined organic phase was washed with water, dried over Na SO , 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 obtain the product.

[0392] C 10 H8Cl2N4O (M=271.1g / mol)

[0393] ESI-MS: 271[M+H] +

[0394] R t (HPLC): 1.01 min (Method B)

[0395] The following compounds were prepared using procedures analogous to those described for Intermediate 1.1 using appropriate starting materials. As will be appreciated by those skilled in the art, these analogous examples may involve variations in the general reaction conditions.

[0396]

[0397]

[0398]

[0399] * p-Anisyl bromide (1.05 eq.) was slowly added to a stirred solution of chloromethyltetrazole and K2CO3 (1.4 eq.) in DMA at 18°C; the mixture was stirred at RT for 1.5 h; purified via reverse phase HPLC (ACN / H2O gradient, 0.1% TFA).

[0400] Intermediate II

[0401] Intermediate II.1 (General Procedure)

[0402] (1R)-2-[5-(Chloromethyl)-2H-1,2,3,4-tetrazolyl-2-yl-]-1-(4-chlorophenyl)ethan-1-ol

[0403]

[0404] Under an inert atmosphere, 1.30 g (4.80 mmol) of 1-(4-chlorophenyl)-2-[5-(chloromethyl)-2H-1,2,3,4-tetrazol-2-yl]ethan-1-one (Intermediate I.1) was dissolved in 20 mL of ACN. 12 mg (0.02 mmol) of ([(1S,2S)-2-amino-1,2-diphenylethyl](4-toluenesulfonyl)amido) chloride was added. Ruthenium(II) (CAS 174813-81-1) was added, followed by the dropwise addition of 0.72 mL (1.73 mmol) of triethylamine formate complex (5:2). After stirring at room temperature for 3 h, the solvent was removed under reduced pressure. Water was added to the remaining crude mixture, and the mixture was extracted with EtOAc. The combined organic layers were dried over Na2SO4, filtered, treated with activated charcoal, filtered, and the solvent removed under reduced pressure to provide Intermediate II.1.

[0405] C 10 H 10 Cl2N4O (M=273.1g / mol)

[0406] ESI-MS: 273[M+H] +

[0407] R t (HPLC): 0.96 min (Method B)

[0408] The following compounds were prepared using procedures analogous to those described for Intermediate II.1 using appropriate starting materials. As will be appreciated by those skilled in the art, these analogous examples may involve variations in the general reaction conditions.

[0409]

[0410]

[0411]

[0412] Intermediate III

[0413] 3-Methyl-2,4-dioxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide

[0414]

[0415] A solution of 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 XII.6) in 33% aqueous ammonia (120 mL) was stirred at 100 ° C in a sealed container for 10 h. The reaction mixture was cooled to RT and concentrated under reduced pressure. The residue was triturated with ACN, filtered off and dried at 50 ° C to obtain intermediate III.

[0416] C6H7N3O3 (M=169.1g / mol)

[0417] ESI-MS: 170[M+H] +

[0418] R t (HPLC): 0.48 min (Method B)

[0419] Intermediate IV

[0420] Intermediate IV.1 (General Procedure)

[0421] 1-(5,6-Difluoro-1-benzofuran-2-yl)ethan-1-one

[0422]

[0423] A solution of 5.00 g (31.6 mmol) of 4,5-difluoro-2-hydroxybenzaldehyde in 50 mL of acetone was treated with 6.99 g (50.6 mmol) of potassium carbonate at 0°C under argon. After stirring for a further 10 min at 0°C, 3.78 mL (47.4 mmol) of chloroacetone was added dropwise and the reaction mixture was stirred at 70°C for 3 h. The reaction mixture was cooled to RT and concentrated. The crude product was extracted with EtOAc / water and the organic phase was concentrated under reduced pressure to give Intermediate IV.1.

[0424] C 10 H6F2O2 (M=196.2g / mol)

[0425] 1 H NMR(300MHz,DMSO-d6)δppm:2.56(s,3H),7.89(m,1H),7.92(m,1H),8.01(m,1H)

[0426] The following compounds were prepared using procedures analogous to those described for Intermediate IV.1 using appropriate starting materials. As will be appreciated by those skilled in the art, these analogous examples may involve variations in the general reaction conditions.

[0427]

[0428]

[0429] Intermediate V

[0430] 5-Bromo-1-benzofuran-2-carboxylic acid

[0431]

[0432] 1.23 g (29.3 mmol) of LiOH * H O was added to a solution of 6.58 g (24.4 mmol) of ethyl 5-bromo-1-benzofuran-2-carboxylate (IV.4) in 3 mL of EtOH, 66 ml of THF, and 33 mL of water at 0° C. The reaction mixture was stirred at RT for 2 h and then concentrated under reduced pressure. The residue was acidified to pH 5 with 1 M HCl, and the resulting precipitate was filtered off and dried to give Intermediate V.

[0433] C9H5BrO3 (M=241.0g / mol)

[0434] 1 H NMR (300MHz, DMSO-d6) δppm: 7.59-7.76 (m, 3H), 8.02 (d, J = 2.0Hz, 1H), 13.5-14.2 (br s, 1H).

[0435] Intermediate VI

[0436] 5-Bromo-2-fluoro-1-benzofuran

[0437]

[0438] A solution of 5.00 g (20.7 mmol) of 5-bromo-1-benzofuran-2-carboxylic acid (V), 14.70 g (41.5 mmol) of Selectflour, and 4.82 g (83.0 mmol) of potassium fluoride in 185 ml of DCE and 95 ml of water was stirred at 70° C. in a sealed tube for 20 h. The reaction mixture was then extracted with DCM / water. The organic layer was washed with brine, dried over Na2SO4, and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel, DCM).

[0439] C8H4BrFO (M=215.0g / mol)

[0440] 1 H NMR (300MHz, DMSO-d6) δppm: 6.36 (dd, J=6.4, 0.9Hz, 1H), 7.47 (dd, J=8.7, 2.1Hz, 1H), 7.58 (d, J=8.7Hz, 1H), 7.82 (d, J=2.1Hz, 1H)

[0441] Intermediate VII

[0442] 1-(2-Fluoro-1-benzofuran-5-yl)ethan-1-one

[0443]

[0444] At RT, 168 mg (1.2 mmol) of potassium carbonate was added to a solution of 218 mg (1.0 mmol) of 5-bromo-2-fluoro-1-benzofuran (VI) in 3 mL of DMF and 0.3 mL of water under stirring. The mixture was purged with argon, followed by the addition of 25 mg (0.1 mmol) of 1,3-bis(diphenylphosphino)propane (dppp), 7 mg of palladium(II) acetate, and 183 mg (2.5 mmol) of ethyl vinyl ether. The reaction mixture was stirred at 80 ° C. overnight, then cooled to RT and treated with a 1 M aqueous solution of HCl (20 mL). After stirring at RT for 30 min, the mixture was extracted with EtOAc and the combined organic layers were concentrated under reduced pressure. The crude product was purified by column chromatography (silica gel; EtOAc / hexane, gradient).

[0445] C 10 H7FO2 (M=178.2g / mol)

[0446] 1 H NMR (300MHz, DMSO-d6) δppm: 2.63 (s, 3H), 6.49 (dd, J=6.4, 0.8Hz, 1H), 7.70 (dt, J=8.7, 0.8Hz, 1H), 7.93 (dd, J=8.7, 1.9Hz, 1H), 8.25 (dd, J=1.9, 0.6Hz, 1H)

[0447] Intermediate VIII

[0448] Intermediate VIII.1 (General Procedure)

[0449] 2-Bromo-1-(2-fluoro-1-benzofuran-5-yl)ethan-1-one

[0450]

[0451] A solution of 126 mg (0.71 mmol) of 1-(2-fluoro-1-benzofuran-5-yl)ethan-1-one (VII) in 1.5 mL of THF was treated dropwise at RT with a solution of 0.34 g (0.71 mmol) of tetrabutylammonium tribromide in 0.08 mL of MeOH and 0.8 mL of THF. After stirring for 2 h, the reaction mixture was concentrated under reduced pressure and the residue was extracted with EtOAc / water. The organic layer was concentrated under reduced pressure and the crude product was purified by column chromatography (silica gel; hexane / EtOAc, gradient).

[0452] C 10 H6BrFO2 (M=257.1g / mol)

[0453] 1 H NMR (300MHz, DMSO-d6) δppm: 4.99 (s, 2H), 6.53 (dd, J=6.4, 0.9Hz, 1H), 7.75 (d, J=8.7, 1H), 7.88-8.03 (m, 1H), 8.31 (dd, J=1.9, 0.6Hz, 1H)

[0454] The following compounds were prepared using procedures analogous to those described for Intermediate VIII.1 using appropriate starting materials. As will be appreciated by those skilled in the art, these analogous examples may involve variations in the general reaction conditions.

[0455]

[0456] *: The reaction was carried out with bromine (13.6 eq) in dioxane / diethyl ether at RT for 2 h and quenched with sodium thiosulfate solution.

[0457] Intermediate IX

[0458] 1-({2-[(2R)-2-(4-chlorophenyl)-2-hydroxyethyl]-2H-1,2,3,4-tetrazolyl-5-yl}methyl)-3-methyl-2,4-dioxo-1,2,3,4-tetrahydropyrimidine-5-carboxylic acid ethyl ester

[0459]

[0460] 303 mg (1.11 mmol) of (1R)-2-[5-(chloromethyl)-2H-1,2,3,4-tetrazol-2-yl]-1-(4-chlorophenyl)ethan-1-ol (Intermediate II.1) and 418 mg (3.03 mmol) of KCO were added to a solution of 200 mg (1.01 mmol) of ethyl 3-methyl-2,4-dioxo-1,2,3,4-tetrahydropyrimidine-5-carboxylate (CAS: 154942-22-0, Intermediate XII.6) in 8 mL of DMF and the mixture was stirred at 50° C. for 5 h and then at RT for 17 h. The crude product was purified by reverse phase HPLC (ACN / HO gradient, 0.1% TFA) to give the desired product.

[0461] C 18 H 19 ClN6O5 (M=434.8g / mol)

[0462] ESI-MS: 435[M+H] +

[0463] R t (HPLC): 0.48 min (Method A)

[0464] Intermediate X

[0465] 1-({2-[(2R)-2-(4-chlorophenyl)-2-hydroxyethyl]-2H-1,2,3,4-tetrazolyl-5-yl}methyl)-3-methyl-2,4-dioxo-1,2,3,4-tetrahydropyrimidine-5-carboxylic acid

[0466]

[0467] 44 mg (1.8 mmol) of lithium hydroxide was added to a solution of 200 mg (0.46 mmol) of intermediate IX in 1 mL of methanol, 1 mL of THF, and 100 μl of water. The reaction mixture was stirred at 50° C. for 1 h and then cooled to RT and diluted with water. The aqueous layer was washed three times with DCM, acidified with formic acid, and the resulting precipitate was filtered off and dried at 50° C. to give the desired product.

[0468] C 16 H 15 ClN6O5 (M=406.8g / mol)

[0469] ESI-MS: 407[M+H] +

[0470] R t (HPLC): 0.46 min (Method A)

[0471] Intermediate XI

[0472] Intermediate XI.1 (General Procedure)

[0473] 1,3-Diethyl 2-{[(cyclobutylcarbamoyl)amino]methylene}malonate

[0474]

[0475] 1.00 g (8.76 mmol) of cyclobutylurea and 3.79 g (17.52 mmol) of 1,3-diethyl 2-(ethoxymethylene)malonate were heated under neat conditions at 100° C. for 2.5 h and at 130° C. for 5 h. The reaction mixture was cooled to RT, diluted with methanol and purified by reverse phase HPLC (ACN / H 2 O gradient, 0.1% TFA) to give intermediate XI.1.

[0476] C 13 H 20 N2O5 (M=284.3g / mol)

[0477] ESI-MS: 285[M+H] +

[0478] R t (HPLC): 0.53 min (Method A)

[0479] The following compounds were prepared using procedures analogous to those described for Intermediate XI.1 using appropriate starting materials. As will be appreciated by those skilled in the art, these analogous examples may involve variations in the general reaction conditions.

[0480]

[0481]

[0482] Intermediate XII

[0483] Intermediate XII.1 (General Procedure)

[0484] 1-Cyclobutyl-2-hydroxy-6-oxo-1,6-dihydropyrimidine-5-carboxylic acid ethyl ester

[0485]

[0486] 957 mg (14.1 mmol) sodium ethoxide are added to a solution of 2.00 g (7.03 mmol) intermediate XI.1 in 30 ml ethanol and the mixture is stirred at 80° C. for 3 h, then diluted with ethanol and purified by reverse phase HPLC (ACN / H 2 O gradient, 0.1% TFA).

[0487] C11 H 14 N2O4 (M=238.2g / mol)

[0488] ESI-MS: 239[M+H] +

[0489] R t (HPLC): 0.37 min (Method A)

[0490] The following compounds were prepared using procedures analogous to those described for Intermediate XII.1 using appropriate starting materials. As will be appreciated by those skilled in the art, these analogous examples may involve variations in the general reaction conditions.

[0491]

[0492]

[0493] Intermediate XII.6

[0494] 3-Methyl-2,4-dioxo-1,2,3,4-tetrahydropyrimidine-5-carboxylic acid ethyl ester

[0495]

[0496] 500 mg (6.75 mmol) of methylurea and 1.36 g (6.75 mmol) of 1,3-diethyl 2-(methoxymethylene)malonate were stirred under neat conditions at 120 ° C for 2 h, at RT for 17 h, at 100 ° C for 66 h, at 150 ° C for 17 h, and at 120 ° C for 17 h. The mixture was then diluted with EtOAc and refluxed. The mixture was slowly cooled to RT and the precipitated intermediate was filtered off.

[0497] C8H 10 N2O4 (M=198.2g / mol)

[0498] ESI-MS: 199[M+H] +

[0499] R t (HPLC): 0.24 min (Method A)

[0500] Intermediate XIII

[0501] Intermediate XIII.1 (General Procedure)

[0502] 1-Cyclobutyl-2-hydroxy-6-oxo-1,6-dihydropyrimidine-5-carboxamide

[0503]

[0504] A solution of 630 mg (0.03 mmol) of intermediate XII.1 in 10 ml of aqueous ammonia solution (33%) was stirred in a sealed container at 85° C. for 17 h. Stirring and addition of aqueous ammonia solution were continued at 100° C. until the starting material was completely consumed. The reaction mixture was then concentrated under reduced pressure to give intermediate XIII.1.

[0505] C9H 11 N3O3 (M=209.2g / mol)

[0506] ESI-MS: 210[M+H] +

[0507] R t (HPLC): 0.31 min (Method A)

[0508] The following compounds were prepared using procedures analogous to those described for Intermediate XIII.1 using appropriate starting materials. As will be appreciated by those skilled in the art, these analogous examples may involve variations in the general reaction conditions.

[0509]

[0510] *Workup: acidify with aqueous HCl (1 M), extract with DCM, concentrate the organic layer under reduced pressure and purify via reverse phase HPLC (ACN / H2O gradient, 0.1% TFA).

[0511] **Purified by reverse phase HPLC

[0512] Intermediate XIV.1

[0513] (Cyclopropylmethyl)urea

[0514]

[0515] 599 mg (7.39 mmol) of potassium cyanate are added to 530 mg (4.93 mmol) of 1-cyclopropylmethylamine hydrochloride in 2 ml of water, and the mixture is stirred for 3 h at 100° C. The reaction mixture is allowed to stand at RT for 14 h, and intermediate XIV.1 is then filtered off.

[0516] C5H 10 N2O (M=114.2g / mol)

[0517] ESI-MS: 115[M+H] +

[0518] R t (HPLC): 0.15 min (Method A)

[0519] Intermediate XIV.2

[0520] (2-Methoxyethyl)urea

[0521]

[0522] 3.24 g (39.94 mmol) potassium cyanate were added in small portions to 2.0 g (26.63 mmol) 2-methoxyethylamine in 8 ml of water and the mixture was stirred at 100° C. for 3 h, cooled to RT, diluted with water / methanol and purified by reverse phase HPLC (ACN / H 2 O gradient, 0.1% TFA).

[0523] C4H 10 N2O2 (M=114.2g / mol)

[0524] ESI-MS: 119[M+H] +

[0525] R t (HPLC): 0.11 min (Method A)

[0526] Preparation of final compound

[0527] Example 1 (General Procedure)

[0528] 1-({2-[(2R)-2-(4-chlorophenyl)-2-hydroxyethyl]-2H-1,2,3,4-tetrazolyl-5-yl}methyl)-3-methyl-2,4-dioxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide

[0529]

[0530] 178 mg (0.65 mmol) of (1R)-2-[5-(chloromethyl)-2H-1,2,3,4-tetrazol-2-yl]-1-(4-chlorophenyl)ethan-1-ol (Intermediate II.1) and 245 mg (1.77 mmol) of KCO were added to a solution of 100 mg (0.59 mmol) of 3-methyl-2,4-dioxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide (Intermediate III) in 5 mL of DMF and the mixture was stirred overnight at RT. The mixture was purified by reverse phase HPLC (ACN / H0 gradient, 0.1% TFA) to give the desired product.

[0531] C 16 H 16 ClN7O4 (M=405.8g / mol)

[0532] ESI-MS: 406[M+H] +

[0533] R t (HPLC): 1.05 min (Method B)

[0534] 1 H NMR (400MHz, DMSO-d6) δppm: 3.21 (s, 3H), 4.73-4.84 (m, 2H), 5.13 (m, 1H), 5.45 (s, 2H), 5.92 (d, J = 2.4Hz, 1H), 7.35-7.42 (m, 4H), 7.64 (br d,J=3.4Hz,1H),8.20(br d,J=3.4Hz,1H),8.77(s,1H)

[0535] The following compounds were prepared using appropriate starting materials using procedures analogous to those described in Example 1. As will be appreciated by those skilled in the art, these analogous examples may involve variations on the general reaction conditions.

[0536]

[0537]

[0538]

[0539]

[0540] The analytical data of the compounds are described in the table above:

[0541]

[0542]

[0543]

[0544]

[0545] Example 22 (General Procedure)

[0546] 1-({2-[(2R)-2-(4-chlorophenyl)-2-hydroxyethyl]-2H-1,2,3,4-tetrazolyl-5-yl}methyl)-N-(2-methoxyethyl)-3-methyl-2,4-dioxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide

[0547]

[0548] Under stirring at RT, 25 μl (0.15 mmol) of DIPEA and 22 mg (0.06 mmol) of HATU were added to a solution of 20 mg (0.05 mmol) of intermediate X in 1.0 mL of DMF. After 30 minutes, 8 μl (0.10 mmol) of 2-methoxyethan-1-amine were added and the mixture was stirred at RT for 90 minutes. Subsequent purification by reverse phase HPLC (ACN / H 2 O gradient, 0.1% TFA) gave the desired product.

[0549] C 19 H 22 ClN7O5 (M=463.9g / mol)

[0550] ESI-MS: 464[M+H] +

[0551] R t (HPLC): 0.48 min (Method A)

[0552] 1 H NMR(400MHz,DMSO-d6)δppm 3.22(s,3H),3.28(s,3H),3.41-3.51(m,4H),4.73-4.84(m,2H),5.13(m,1H),5.45(s,2H),5.5-6.4(br s,1H),7.38(m,4H),8.77(s,1H),8.94(m,1H).

[0553] The following compounds were prepared using appropriate starting materials using procedures analogous to those described in Example 22. As will be appreciated by those skilled in the art, these analogous examples may involve variations on the general reaction conditions.

[0554]

[0555]

[0556]

[0557]

[0558] The analytical data of the compounds are described in the table above:

[0559]

[0560]

[0561]

[0562] Analytical HPLC method

[0563] Method A

[0564]

[0565] Analytical column: XBridge BEH (Waters) C18_2.1×30mm_1.7μm; column temperature: 60℃

[0566] Method B

[0567]

[0568] Analytical column: Stable Bond (Agilent) C18_3.0×30mm_1.8μm; column temperature: 60℃

[0569] Method C

[0570]

[0571] Analytical column: XBridge (Waters) C18_3.0×30mm_2.5μm; column temperature: 60℃

[0572] Method D

[0573]

[0574] Analytical column: XBridge C18_3.0×30mm_2.5μm (Waters); column temperature: 60℃

[0575] Method E

[0576]

[0577] Analytical column: Sunfire (Waters); C18_3.0×30mm_2.5μm; column temperature: 60°C

[0578] Method F

[0579]

[0580] Analytical column: XBridge BEH (Waters) C18_2.1×30mm_2.5μm; column temperature: 60℃

[0581] Method G

[0582]

[0583]

[0584] Analytical column: Zorbax StableBond C18 (Agilent) 1.8 μm; 2.1 × 30 mm; column temperature: 60 °C

[0585] Method H

[0586]

[0587] Analytical column: Sunfire (Waters) 2.5 μm; 3.0 × 30 mm; column temperature: 60 °C

[0588] Method I

[0589]

[0590] Analytical column: Sunfire C18 (Waters) 2.5 μm; 3.0 × 30 mm; column temperature: 60 °C

[0591] Method J

[0592]

[0593] Analytical column: Acquity UPLC BEH; C8_2.1×150mm_1.7μm; column temperature: 55°C.

Claims

1. A compound according to formula (I) in A is selected from the group consisting of: R 1 Selected from the group consisting of: H3C、H3CH2C、H3COH2CH2C、 R 2 Selected from the group consisting of: H, H3C, H3CH2C, H3COH2CH2C, F2HCH2C, F3CH2C, FH2CH2C, H3C(O)2SH2CH2C, F3COH2CH2C, 2. A compound of formula (I) according to claim 1 , which is selected from the group consisting of:

3. A compound of formula (I) according to claim 1, wherein R 1 It’s H3C.

4. A compound of formula (I) according to claim 1, wherein R 2 It’s H.

5. A compound selected from the group consisting of:

6. A compound which is 7. A compound which is 8. A compound which is 9. A compound which is 10. A compound which is 11. A compound which is 12. A compound which is 13. A compound which is 14. A compound which is 15. A compound which is 16. A salt of a compound according to any one of claims 1 to 15.

17. A pharmaceutical composition comprising at least one compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients.

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