Modulators of intracellular chloride concentration

Abstract

Modulators of intracellular chloride concentration are disclosed. The present invention relates in particular to compounds of Formula Ia, Ib and Ic, pharmaceutical compositions comprising the compounds of Formula Ia, and their use in the treatment or prevention of pathological conditions associated with the delivery of depolarized GABA, including, for example, Down's disease and autism.

Description

[0001] This application is a divisional application of Chinese patent application No. 202080041313.5, entitled “Regulators of intracellular chloride concentration”, filed on April 2, 2020.

[0002] Citation of Related Applications

[0003] This patent application claims the benefit of Italian patent application no. 102019000004929, filed on April 2, 2019, the disclosure of which is incorporated herein by reference in its entirety. Technical Field

[0004] The present invention relates to compounds of formula Ia, Ib and Ic which inhibit the sodium, potassium and chloride cotransporter (hereinafter also referred to as NKCC1).

[0005]

[0006] Pharmacological inhibition of NKCC1 could be used to treat a variety of pathophysiological conditions, particularly brain disorders. 2-Aminobenzenesulfonamide derivatives are potent NKCC1 inhibitors and have shown promising efficacy in restoring GABAergic transmission and associated cognitive behaviors in rodent models of Down syndrome and autism. Background Art

[0007] Down syndrome is the most common inherited form of intellectual disability (occurring in approximately 10 and 14 per 10,000 live births in European countries and the United States, respectively). Down syndrome, also known as trisomy 21, is a genetic condition caused by the presence of all or part of the third copy of chromosome 21. The most prominent clinical feature of Down syndrome is intellectual disability, manifested by low IQ, learning deficits, and memory impairment, particularly in hippocampal function. Although teaching methods and mainstreaming education have improved cognitive development in individuals with Down syndrome, constitutive deficits remain that these approaches cannot fully address. Indeed, despite the existence of multiple clinical candidates for the treatment of Down syndrome (i.e., piracetam, memantine, and donepezil, rivastigmine, epigallocatechin gallate and antioxidants, pentylenetetrazol, and ACI-24), there are currently no approved drugs for improving cognitive symptoms in Down syndrome. Therefore, efforts to identify drugs that enhance cognitive function in individuals with Down syndrome are urgently needed.

[0008] In the past few years, a large number of literatures have shown that in Down syndrome and many other neurodevelopmental diseases, inhibition of GABAergic transmission through Cl-permeable GABAA receptors is defective (Deidda, G. et al. Modulation of GABAergic transmission in development and neurodevelopmental disorders: investigating physiology and pathology to gain therapeutic perspectives. Front Cell Neurosci 2014, 8, 119.3; Contestabile, A. et al. The GABAergic Hypothesis for Cognitive Disabilities in Down syndrom. Frontiers in Cellular Neurosciences 2017, 11.54). However, using common GABAA receptor inhibitors to restore defective GABAergic transmission is dangerous. This is because patients are at high risk of epileptic seizures.

[0009] Brain diseases characterized by altered GABAergic transmission include Down syndrome, neuropathic pain, stroke, cerebral ischemia, cerebral edema, hydrocephalus, traumatic brain injury, depressive-like behavior induced by brain injury, autism spectrum disorders (i.e., autism, fragile X, Rett syndrome, Asperger's and DiGeorge syndromes), epilepsy, seizures, status epilepticus, childhood spasms, gliomas, glioblastomas, anaplastic astrocytomas, Parkinson's disease, Huntington's disease, schizophrenia, anxiety, tuberous sclerosis and related behavioral problems, and Dravet syndrome. Na is encoded by the SLC12A2 (NKCC1) and SLC12A1 (NKCC2) genes. + , K + 、Cl - The NKCC cotransporters belong to a family of transporters that provide electroneutral transport of sodium, potassium, and chloride across the plasma membrane; they move each solute in the same direction and maintain electroneutrality by moving two positively charged solutes (sodium and potassium) along with two partially negatively charged solutes (chloride).

[0010] NKCC1 is widely distributed, especially in the exocrine glands and brain; NKCC2 is found in the kidneys and is used to extract sodium, potassium, and chloride from the urine so that they can be reabsorbed into the blood. - Input NKCC1 and Cl- The exporter KCC2 mainly controls intracellular Cl - concentration.

[0011] Importantly, the NKCC1 / KCC2 expression ratio is defective in animal models of Down syndrome and various brain diseases; targeting NKCC1 with inhibitors has therapeutic effects on a variety of diseases, including but not limited to Down syndrome, neuropathic pain, stroke, cerebral ischemia, cerebral edema, hydrocephalus, traumatic brain injury, depressive-like behavior induced by brain trauma, autism spectrum disorders (i.e., autism, fragile X, Rett syndrome, Asperger and DiGeorge syndromes), epilepsy, convulsions, status epilepticus, childhood spasms, gliomas, glioblastomas, anaplastic astrocytomas, Parkinson's disease, Huntington's disease, schizophrenia, anxiety, tuberous sclerosis and related behavioral problems, Dravet syndrome. In animal models, inhibition of NKCC1 by the FDA-approved diuretic bumetanide rescues behavioral deficits. Notably, bumetanide restores GABAAR-driven Cl in adult Down syndrome mouse models. - Currents, synaptic plasticity, and hippocampus-dependent memory. Therefore, NKCC1 inhibitors have been shown to have therapeutic activity in diseases in which GABAergic transmission is defective.

[0012] Furthermore, in five separate clinical studies (including a Phase II clinical trial), bumetanide treatment reduced ratings and perception of emotional faces in children with autism.

[0013] However, bumetanide has a diuretic effect because it also inhibits renal-specific Cl - This diuretic effect can cause ion imbalance and seriously compromise medication compliance during long-term treatment.

[0014] Conditions for which bumetanide has been shown to improve include Down syndrome, neuropathic pain, stroke, cerebral ischemia, cerebral edema, hydrocephalus, traumatic brain injury, depressive-like behavior induced by brain injury, autism spectrum disorders (i.e., autism, fragile X, Rett syndrome, Asperger's, and DiGeorge syndromes), epilepsy, convulsions, status epilepticus, childhood spasms, gliomas, glioblastomas, anaplastic astrocytomas, Parkinson's disease, Huntington's disease, schizophrenia, anxiety, tuberous sclerosis and related behavioral problems, and Dravet syndrome.

[0015] WO 2010 / 085352 describes the use of NKCC1 modulators to improve cognitive function in subjects in need. These compounds are also said to be useful for long-term treatment due to reduced undesirable diuresis. The most promising compound, 3-aminosulfonyl-5-N,N-dibutylamino-4-phenoxybenzoic acid, is described as interacting with GABAA receptors, and therefore is neither an NKCC1 nor an NKCC2 inhibitor, and may carry the risk of undesirable side effects, including epileptic seizures.

[0016] WO 2014 / 076235 describes compounds for the treatment of Fragile X syndrome. In a preferred embodiment, the chloride modulator is a selective inhibitor of NKCC1.

[0017] In a publication by Huang et al. (“Novel NKCC1 Inhibitors Reduces Stroke Damages”; Stroke, April 2019), the power of STS66, 3-(butylamino)-2-phenoxy-benzenesulfonamide, was investigated. This compound is a close analog and derivative of bumetanide and therefore acts as an NKCC1 inhibitor.

[0018] Lykke et al., in "The search for NKCC1-selective drugs for the treatment of epilepsy: Structure-function relationship of bumetanide and various bumetanide derivatives in inhibiting the human cation-chloride cotransporter NKCC1A," Epilepsy & Behavior 59 (2016) 42-49, investigated bumetanide derivatives as selective inhibitors of NKCC1. Test derivatives were selected from approximately 5,000 3-amino-5-sulfamoylbenzoic acid derivatives synthesized by Peter W. Feit and colleagues at Leo Pharma in the 1960s and 1970s during a screen for compounds with high diuretic potency, ultimately leading to the discovery of bumetanide. According to the authors, none of the compounds exhibited significantly higher NKCC2 / NKCC1 selectivity. The authors concluded that developing bumetanide derivatives with higher selectivity for NKCC1 versus NKCC2 than bumetanide would be difficult, if not impossible.

[0019] Therefore, there is a need for alternative therapeutic approaches for Down syndrome and other brain disorders that would enable restoration of defective GABAergic transmission by inhibiting NKCC1.

[0020] Therefore, bumetanide is not a viable treatment strategy, and the same is true for the described analogs. There remains a strong need for alternative compounds. Summary of the Invention

[0021] The present invention relates to novel 2-aminobenzenesulfonamide derivatives that inhibit the sodium, potassium, and chloride cotransporter (also referred to herein as NKCC1). Pharmacological inhibition of NKCC1 can be used to treat a variety of pathophysiological conditions, particularly brain disorders. Modulation of NKCC1 leads to fine-tuning of GABAergic transmission. Thus, NKCC1 inhibitors are effective in treating cells with defective NKCC1 / KCC2 expression ratios and / or via Cl - It has beneficial effects in diseases characterized by defective GABAergic transmission at permeable GABAA receptors.An object of the present invention is to treat diseases involving GABAA and / or chloride homeostasis.

[0022] Purpose of the Invention

[0023] According to the first object, the present invention provides novel 2-aminobenzenesulfonamide derivatives that are capable of inhibiting the sodium, potassium and chloride cotransporter (also referred to as NKCC1).

[0024] The present invention also discloses a method for preparing the disclosed compound.

[0025] In a second object, the use of the compounds of the present invention for the treatment or prevention of pathological conditions associated with depolarization of GABAergic transmission is disclosed.

[0026] Pharmaceutical formulations comprising the compounds of the invention represent a third object of the present invention.

[0027] In a fourth object, there is disclosed a method for treating or preventing a pathological condition associated with depolarization of GABAergic transmission, comprising administering a compound of the present invention to a patient in need thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1: In vitro testing of NKCC1 inhibitors in a chloride kinetics assay. a) Example traces obtained in a chloride kinetics assay on HER cells transfected with YFP (mock) or with both YFP and NKCC1. Arrows indicate the addition of NaCl (final concentration 74 mM) to initiate the flux assay. b) Quantification of the effects of bumetanide (10 μM and 100 μM) or furosemide (10 μM and 100 μM) on mock- or NKCC1-transfected HEK293 cells in a chloride kinetics assay. Data represent mean ± sem from five independent experiments. c) Quantification of the effects of bumetanide and furosemide, as well as two selected compounds (3.8 and 3.17), on NKCC1-transfected HEK293 cells in a chloride kinetics measurement. Data represent mean ± sem from five independent experiments and are expressed as % of control. *P<0.05, **P<0.01, ***P<0.001, Kruskal-Wallis ANOVA with Dunn's post hoc test; ###P<0.001, two-tailed unpaired Student's t-test.

[0029] Figure 2 NKCC1 inhibitors were tested in vitro in a calcium dynamics assay. a) Example traces of fluorescence levels following GABA (100 μM) and KCl (90 mM) administration to trigger calcium influx in treated primary neuronal cultures after 3 days of culture (3 DIV) with vehicle, bumetanide, furosemide, and compounds 3.8, 3.13, and 3.17 in a calcium dynamics assay. b) Quantification of the mean fluorescence increase following GABA application, normalized to the increase following KCl application, in neurons treated with bumetanide, furosemide, and three exemplary compounds (3.8, 3.13, 3.17) (10 μM, 100 μM). Data represent mean ± sem from five independent experiments and are expressed as % of control. *P < 0.05, **P < 0.01, ***P < 0.001, Kruskal-Wallis ANOVA with Dunn's post hoc test.

[0030] Figure 3 : Pharmacokinetic evaluation of selected compounds, Compound 3.17. a) Physicochemical properties of bumetanide and Compound 3.17 analyzed by LC-MS. b) Comparison of urine output in WT (C57Bl / 6N) mice after two-hour treatment with bumetanide (0.2 mg / kg) and Compound 3.17 (0.2 mg / kg). c) Evaluation of urine output in Down syndrome and WT littermate Ts65Dn mice after two-hour treatment with Compound 3.17 (0.2 mg / kg). Numbers in parentheses: number of animals analyzed. Data represent mean ± sem and are expressed as % of the corresponding vehicle.

[0031] Figure 4In vivo evaluation of the efficacy of selected NKCC1 inhibitors in Ts65Dn mice. (a) Quantification of the discrimination index in mice treated with vehicle (WT, n=14, Ts65Dn, n=10) or 3.17 (WT, n=14, Ts65Dn, n=11), ***P<0.001; two-way ANOVA with Tukey's post hoc test. (b) Quantification of the discrimination index in mice treated with vehicle (WT, n=14, Ts65Dn, n=10) or 3.17 (WT, n=14, Ts65Dn, n=11), *P<0.05, **P<0.01, two-way ANOVA with Tukey's post hoc test. (c) Quantification of normal choice in mice treated with vehicle (WT, n=14, Ts65Dn, n=10) or 3.17 (WT, n=14, Ts65Dn, n=11), ***P<0.001; two-way ANOVA with Tukey's post hoc test. (d) Quantification of freezing responses in mice treated with vehicle (WT, n=14, Ts65Dn, n=10) or 3.17 (WT, n=14, Ts65Dn, n=11), *P<0.05, **P<0.01, two-way ANOVA with Tukey's post hoc test.

[0032] Figures 5 to 16 : Synthetic procedures Schemes 1 to 15 for the preparation of compounds of the present invention are reported.

[0033] Figure 17 : Shows the results of in vitro testing of selective NKCC1 inhibitors in a thallium-based assay on NKCC2-transfected HER cells.

[0034] Figure 18: Shows the results of in vivo evaluation of the efficacy of compound 3.17 in a VPA-induced autism mouse model; (a) Left panel, quantification of the social index in mice treated with vehicle (WT, n=15, VPA, n=10) or 3.17 (WT, n=9, VPA, n=12); two-way ANOVA on ranks, Tukey post hoc test, **P<0.01. Right panel, quantification of the social novelty index in mice treated with vehicle (WT, n=15, VPA, n=10) or 3.17 (WT, n=9, VPA, n=12); two-way ANOVA, Tukey post hoc test, *P<0.05, **P<0.01. (b) Quantification of interaction time in mice treated with vehicle (WT, n = 15, VPA, n = 10) or 3.17 (WT, n = 10, VPA, n = 11); two-way ANOVA, Tukey's post hoc test, **P < 0.01. (c) Quantification of the number of marbles buried by mice treated with vehicle (WT, n = 17, VPA, n = 17) or 3.17 (WT, n = 13, VPA, n = 13); two-way ANOVA, Tukey's post hoc test, *P < 0.05, **P < 0.01. (d) Quantification of grooming time in mice treated with vehicle (WT, n = 20, VPA, n = 17) or 3.17 (WT, n = 13, VPA, n = 13); two-way ANOVA on ranks, Tukey's post hoc test, *P < 0.05, **P < 0.01. DETAILED DESCRIPTION

[0035] The present invention provides 2-aminobenzenesulfonamide derivatives according to Formula Ia, Ib and Ic that are NKCC1 inhibitors and address the need for alternative compounds to bumetanide, and in particular compounds that are capable of restoring GABA A signaling through NKCC1 inhibition.

[0036] In one aspect, the present invention provides a compound having Formula Ia or a pharmaceutically acceptable salt thereof or a stereoisomeric form thereof, or individual geometric isomers, enantiomers, diastereomers, tautomers, zwitterions and pharmaceutically acceptable salts thereof:

[0037]

[0038] in:

[0039] R1 and R2 are independently

[0040] ·hydrogen;

[0041] ·Straight chain or branched chain C 1-10 Alkyl, optionally containing one or more unsaturations and optionally substituted by halogen, -OH, -C 3-8Cycloalkyl, non-aromatic heterocycle, aromatic heterocycle, -C 1-6 Alkoxyalkyl, -NH2, -NO2, amide, carboxylic acid, ketone, ether, ester, aldehyde or sulfonamide;

[0042] · Straight-chain or branched, substituted or unsubstituted C 3-8 Cycloalkyl;

[0043] · Straight-chain or branched, substituted or unsubstituted C 4-10 cycloalkylalkyl;

[0044] ·C 3-8 heterocycloalkyl;

[0045] optionally substituted phenyl;

[0046] or R1 and R2 together with the nitrogen atom to which they are attached form a substituted or unsubstituted saturated heterocyclic ring;

[0047] R3 and R4 are independently

[0048] ·hydrogen;

[0049] ·Straight chain or branched chain C 1-10 Alkyl, optionally containing one or more unsaturations and optionally substituted by halogen, -OH, -C 3-8 Cycloalkyl, non-aromatic heterocycle, aromatic heterocycle, -C 1-6 Alkoxyalkyl, -NH2, -NO2, amide, carboxylic acid, ketone, ether, ester, aldehyde or sulfonamide;

[0050] ·C 3-10 Cycloalkyl;

[0051] ·C 4-10 cycloalkylalkyl;

[0052] ·C 2-8 alkyl halide;

[0053] ·Straight chain or branched chain C 2-8 heteroalkyl, substituted or unsubstituted;

[0054] optionally substituted phenyl;

[0055] Provided that at least one of R3 and R4 is not hydrogen;

[0056] or R3 and R4, when taken together with the nitrogen atom to which they are attached, form a substituted or unsubstituted saturated heterocyclic ring;

[0057] R5 is

[0058] ·hydrogen;

[0059] ·halogen;

[0060] hydroxyl groups;

[0061] -OC 1-10 alkyl;

[0062] -OC 3-10 Cycloalkyl;

[0063] -OC 3-8 heterocycloalkyl;

[0064] ·C 1-10 Alkoxyalkyl;

[0065] ·C 3-10 Alkoxycycloalkyl;

[0066] optionally substituted phenoxy;

[0067] -NH2;

[0068] ·C 1-8 Alkylamines;

[0069] C2-C 16 dialkylamines;

[0070] ·aniline;

[0071] -SH;

[0072] ·C 1-8 Alkyl sulfides;

[0073] Thiophenol;

[0074] -NO2;

[0075] R6 is

[0076] Nitro;

[0077] Nitrile;

[0078] -CH2OH;

[0079] ·carboxylic acid;

[0080] ·C 1-4 alkyl esters;

[0081] ·C 2-8 heteroalkyl esters;

[0082] ·C 3-6 Cycloalkyl esters;

[0083] Phenyl esters;

[0084] Carboxamides;

[0085] Cyclic amides;

[0086] Tetrazole;

[0087] The condition is that when R6 is nitro, the following conditions are met at the same time:

[0088] R1 is not H,

[0089] R2 is not a linear or branched, unsubstituted C 2-6 alkyl,

[0090] R3 is not H,

[0091] R4 is not a linear unsubstituted C 1-3 alkyl,

[0092] R5 is not H;

[0093] and with the proviso that the compound of Formula Ia is not one of the following:

[0094]

[0095]

[0096] In one embodiment:

[0097] R1 and R2 are independently

[0098] ·hydrogen;

[0099] ·Straight chain or branched chain C 1-10 Alkyl, optionally containing one or more unsaturations and optionally substituted by halogen, -OH, -C 3-8 Cycloalkyl, non-aromatic heterocycle, aromatic heterocycle, -C 1-6 Alkoxyalkyl, -NH2, -NO2, amide, carboxylic acid, ketone, ether, ester, aldehyde or sulfonamide;

[0100] · Straight-chain or branched, substituted or unsubstituted C 3-8 Cycloalkyl;

[0101] · Straight-chain or branched, substituted or unsubstituted C 4-10 cycloalkylalkyl;

[0102] optionally substituted phenyl;

[0103] or R1 and R2 together with the nitrogen atom to which they are attached form a substituted or unsubstituted saturated heterocyclic ring;

[0104] R3 and R4 are independently

[0105] ·hydrogen;

[0106] ·Straight chain or branched chain C 1-10Alkyl, optionally containing one or more unsaturations and optionally substituted by halogen, -OH, -C 3-8 Cycloalkyl, non-aromatic heterocycle, aromatic heterocycle, -C 1-6 Alkoxyalkyl, -NH2, -NO2, amide, carboxylic acid, ketone, ether, ester, aldehyde or sulfonamide;

[0107] ·C 3-10 Cycloalkyl;

[0108] ·C 4-10 cycloalkylalkyl;

[0109] ·C 2-8 alkyl halide;

[0110] ·Straight chain or branched chain C 2-8 heteroalkyl, substituted or unsubstituted;

[0111] optionally substituted phenyl;

[0112] Provided that at least one of R3 and R4 is not hydrogen;

[0113] or R3 and R4, when taken together with the nitrogen atom to which they are attached, form a substituted or unsubstituted saturated heterocyclic ring;

[0114] R5 is

[0115] ·hydrogen;

[0116] ·halogen;

[0117] hydroxyl groups;

[0118] ·C 1-10 Alkoxyalkyl;

[0119] ·C 3-10 Alkoxycycloalkyl;

[0120] optionally substituted phenoxy;

[0121] -NH2;

[0122] ·C 1-8 Alkylamines;

[0123] C2-C 16 dialkylamines;

[0124] ·aniline;

[0125] -SH;

[0126] ·C 1-8 Alkyl sulfides;

[0127] Thiophenol;

[0128] -NO2;

[0129] R6 is

[0130] Nitro;

[0131] Nitrile;

[0132] -CH2OH;

[0133] ·carboxylic acid;

[0134] ·C 1-4 alkyl esters;

[0135] ·C 2-8 heteroalkyl esters;

[0136] ·C 3-6 Cycloalkyl esters;

[0137] Phenyl esters;

[0138] Carboxamides;

[0139] Cyclic amides;

[0140] Tetrazole.

[0141] In a preferred embodiment, R1 and R2 are independently H, -CH3, cyclopentane, cyclohexane, 4-tetrahydropyran, or together with the nitrogen atom to which they are attached, morpholine, piperidine optionally substituted with at least one halogen, pyrrolidine. Still more preferably, R1 and R2 are independently -CH3, -C2H5, -C3H7, -C4H9. In a preferred embodiment, R1 and R2 are both -CH3.

[0142] In a preferred embodiment, R3 and R4 are independently hydrogen, optionally replaced by at least one C 1-6 Alkoxyalkyl substituted straight or branched chain -C 1-8 Alkyl, -C 2-8 Haloalkyl, or R3 and R4, when taken together with the nitrogen atom to which they are attached, are substituted or unsubstituted saturated heterocyclic rings. Still more preferably, R3 and R4 are independently H, -C4H9, -C6H 13 、-C8H 17 、-C2H4C(CH3)3、-C7H 14 CF3, -C3H6CF3, -C5H 10 CF3, -C2H4OCH3, -C4H8OCH3, -C6H 12 OCH3, or together with the nitrogen atom to which they are attached, piperazine, preferably substituted piperazine, still more preferably -N(C4H8CF 3) Piperazine.

[0143] Still more preferably, R3 and R4 are independently -CH3, -C2H5, -C3H7, -C4H9, -C3H 11 、-C6H 13 、-C7H 15 、-C8H 17 or -C 1-8 In a preferred embodiment, R3 is H and R4 is -C7H 14 CF3.

[0144] For the purposes of the present invention, one or more of the hydrogen atoms of the compounds detailed above may be replaced by deuterium.

[0145] In a preferred embodiment, R5 is hydrogen, halogen or hydroxy, more preferably hydrogen.

[0146] In a preferred embodiment, R6 is a carboxylic acid, C 1-4 Alkyl ester, nitro or nitrile, more preferably carboxylic acid.

[0147] In one embodiment, the claimed compound is compound 3.17, having the formula reported below.

[0148]

[0149] definition

[0150] Unless otherwise stated in this specification, it should be understood that the terms used herein have the following meanings.

[0151] As used herein, the term "alkyl" as a single substituent or as part of a larger substituent refers to a saturated monovalent or divalent hydrocarbon moiety having straight or branched chain portions or combinations thereof and containing 1 to 10, preferably 1 to 8, and still more preferably 1 to 4 carbon atoms. Suitable examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, 2-methylbutyl, neopentyl, 1-ethylpropyl, n-hexyl, isohexyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, 2-ethylbutyl, 1-methyl-2-methylpropyl, and the like. The hydrogen atoms on the alkyl group may be substituted by groups including but not limited to: deuterium, halogen, -OH, -C 3-8 Cycloalkyl, non-aromatic heterocycle, aromatic heterocycle, -C 1-6 Alkoxyalkyl, -NH2, -NO2, amide, carboxylic acid, ketone, ether, ester, aldehyde or sulfonamide. For the purposes of the present invention, an alkyl substituent may contain one or more degrees of unsaturation.

[0152] As used herein, the term "cycloalkyl" refers to a monovalent or divalent ring of 3 to 10 carbon atoms or 3 to 8 carbon atoms derived from a saturated cyclic hydrocarbon. Cycloalkyl groups can be monocyclic or polycyclic. Cycloalkyl groups can be substituted with groups including, but not limited to, halogen, -OH, -C 3-8 Cycloalkyl, non-aromatic heterocycle, aromatic heterocycle, -C 1-6 Alkoxyalkyl, -NH2, -NO2, amide, ether, ester, carboxylic acid, aldehyde, ketone, sulfonamide groups.

[0153] Examples of cycloalkylalkyl groups include cyclobutylethyl, cyclobutylpropyl, cyclopentylmethyl, cyclopentylethyl, cyclopentylpropyl, cyclohexylmethyl, cyclohexylethyl, cyclohexylpropyl, cycloheptylmethyl, and cycloheptylethyl.

[0154] As used herein, the term "haloalkyl" refers to an alkyl group that is partially or fully substituted by the same or different halogen atoms. Examples of "haloalkyl" include -CH2CF3 and -CCl2CF3.

[0155] In the present invention, the “alkyloxy” includes, for example, the aforementioned alkyl-O- group, and for example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy and the like can be mentioned, and “alkoxyalkyl” is, for example, methoxymethyl and the like, and “aminoalkyl” is, for example, 2-aminoethyl and the like.

[0156] In the present invention, "halogen" refers to any halogen element, such as fluorine, chlorine, bromine or iodine.

[0157] As used herein, the term "heterocycle" refers to a 3- to 8-membered ring that may be aromatic or non-aromatic, containing at least one heteroatom selected from O, N, or S, or a combination of at least two thereof, that interrupts the carbocyclic ring structure. The heterocyclic ring may contain C=O; the S heteroatom may be oxidized. The heterocyclic ring may be monocyclic or polycyclic. The heterocyclic ring portion may be substituted with groups including, but not limited to, halogen, -OH, -C 1-10 Alkyl, -C 3-8 Cycloalkyl, non-aromatic heterocycle, aromatic heterocycle, -C 1-6 Alkoxyalkyl, -NH2, -NO2, amide, ether, ester, aldehyde, carboxylic acid, ketone, sulfonamide group. Preferred heterocycles are aziridine, azetidine, pyrrolidine, imidazoline, pyrazoline, piperidine, piperazine, morpholine, thiomorpholine, azepane, azooctane.

[0158] As used herein, the term "substituted heterocycle" refers to a ring optionally substituted with halogen, -C 1-5 Alkyl, -C 1-5 Alkenyl, -C 1-5 Heterocyclic ring substituted with haloalkyl.

[0159] As used herein, the term "alkenyl" refers to a monovalent or divalent hydrocarbon radical having 2 to 6 carbon atoms and having at least one double bond, derived from a saturated alkyl group. 2-6 The alkenyl group may be in the E or Z configuration. 1-6 Alkyl substitution.

[0160] As used herein, "substituted phenyl" or "substituted phenoxy" refers to a substituted phenyl group selected from C 1-8 Alkyl (preferably methyl), C 1-8 Phenyl substituted with a substituent selected from the group consisting of alkoxy (preferably methoxy), hydroxy, trifluoromethyl, nitro, amine, and halogen.

[0161] The term "pharmaceutically acceptable salt" refers to a salt or complex that retains the desired biological activity of the above-mentioned compound and exhibits minimal or no undesirable toxicological effects. "Pharmaceutically acceptable salts" according to the present invention include therapeutically active, non-toxic base or acid salt forms that the compound of formula I is able to form.

[0162] The compound of formula Ia and its salt may be in the form of a solvate, which is included within the scope of the present invention. Such solvates include, for example, hydrates, alcoholates, and the like.

[0163] With respect to the present invention, a reference to one or more compounds is intended to encompass each compound in its possible isomeric forms and mixtures thereof, unless specific isomeric forms are specifically mentioned.

[0164] The compounds according to the present invention may exist in different polymorphic forms; although not explicitly indicated in the above formula, these forms are intended to be included within the scope of the present invention.

[0165] In one embodiment, the compound of Formula Ia is selected from the group consisting of:

[0166] 1.6 2-(Butylamino)-5-nitro-benzenesulfonamide,

[0167] 1.7 2-(Hexylamino)-5-nitrobenzenesulfonamide,

[0168] 1.8 5-Nitro-2-(octylamino)benzenesulfonamide,

[0169] 1.9 2-(3,3-dimethylbutylamino)-5-nitro-benzenesulfonamide,

[0170] 1.10 2-(Butylamino)-N-methyl-5-nitrobenzenesulfonamide,

[0171] 1.11 2-(Hexylamino)-N-methyl-5-nitrobenzenesulfonamide,

[0172] 1.12 N-methyl-5-nitro-2-(octylamino)benzenesulfonamide,

[0173] 1.13 2-(3,3-Dimethylbutylamino)-N-methyl-5-nitrobenzenesulfonamide,

[0174] 1.14 2-(Butylamino)-N,N-dimethyl-5-nitrobenzenesulfonamide,

[0175] 1.15 2-(Hexylamino)-N,N-dimethyl-5-nitrobenzenesulfonamide,

[0176] 1.16 N,N-dimethyl-5-nitro-2-(octylamino)benzenesulfonamide,

[0177] 1.17 2-(3,3-Dimethylbutylamino)-N,N-dimethyl-5-nitrobenzenesulfonamide,

[0178] 2.2 4-(Butylamino)-2-chloro-5-sulfamoyl-benzoic acid,

[0179] 2.3 2-Chloro-4-(hexylamino)-5-sulfamoyl-benzoic acid,

[0180] 2.4 2-Chloro-4-(octylamino)-5-sulfamoyl-benzoic acid,

[0181] 2.5 2-Chloro-4-(3,3-dimethylbutylamino)-5-sulfamoyl-benzoic acid,

[0182] 2.6 4-(Butylamino)-3-sulfamoyl-benzoic acid,

[0183] 2.7 4-(Hexylamino)-3-sulfamoyl-benzoic acid,

[0184] 2.8 4-(Octylamino)-3-sulfamoyl-benzoic acid,

[0185] 2.9 4-(3,3-Dimethylbutylamino)-3-sulfamoyl-benzoic acid,

[0186] 3.6 4-(Butylamino)-3-(methylsulfamoyl)benzoic acid,

[0187] 3.7 4-(Hexylamino)-3-(methylsulfamoyl)benzoic acid,

[0188] 3.8 3-(Methylsulfamoyl)-4-(octylamino)benzoic acid,

[0189] 3.9 4-(3,3-dimethylbutylamino)-3-(methylsulfamoyl)benzoic acid,

[0190] 3.10 3-(Methylsulfamoyl)-4-(8,8,8-trifluorooctylamino)benzoic acid,

[0191] 3.11 4-(Butylamino)-3-(dimethylsulfamoyl)benzoic acid,

[0192] 3.12 3-(Dimethylsulfamoyl)-4-(hexylamino)benzoic acid,

[0193] 3.13 3-(Dimethylsulfamoyl)-4-(octylamino)benzoic acid,

[0194] 3.14 4-(3,3-Dimethylbutylamino)-3-(dimethylsulfamoyl)benzoic acid,

[0195] 3.15 3-(Dimethylsulfamoyl)-4-(4,4,4-trifluorobutylamino)benzoic acid,

[0196] 3.16 3-(Dimethylsulfamoyl)-4-(6,6,6-trifluorohexylamino)benzoic acid,

[0197] 3.17 3-(Dimethylsulfamoyl)-4-(8,8,8-trifluorooctylamino)benzoic acid,

[0198] 3.18 3-(Dimethylsulfamoyl)-4-(2-methoxyethylamino)benzoic acid,

[0199] 3.19 3-(Dimethylsulfamoyl)-4-(4-methoxybutylamino)benzoic acid,

[0200] 3.20 3-(Dimethylsulfamoyl)-4-(6-methoxyhexylamino)benzoic acid,

[0201] 3.21 3-(Cyclopentylsulfamoyl)-4-(8,8,8-trifluorooctylamino)benzoic acid,

[0202] 3.22 3-(Cyclohexylsulfamoyl)-4-(8,8,8-trifluorooctylamino)benzoic acid,

[0203] 5.5 3-Pyrrolidin-1-ylsulfonyl-4-(8,8,8-trifluorooctylamino)benzoic acid,

[0204] 5.6 3-(1-piperidinylsulfonyl)-4-(8,8,8-trifluorooctylamino)benzoic acid,

[0205] 5.7 3-Morpholinosulfonyl-4-(8,8,8-trifluorooctylamino)benzoic acid,

[0206] 6.3 5-Cyano-N,N-dimethyl-2-(8,8,8-trifluorooctylamino)benzenesulfonamide,

[0207] 7.4 2-Hydroxy-5-sulfamoyl-4-(8,8,8-trifluorooctylamino)benzoic acid,

[0208] 9.1 3-(Dimethylsulfamoyl)-4-[4-(5,5,5-trifluoropentyl)piperazin-1-yl]benzoic acid,

[0209] 10.1N,N-dimethyl-5-(1H-tetrazol-5-yl)-2(8,8,8-trifluorooctylamino)benzenesulfonamide,

[0210] 12.3 Methyl 5-(N,N-dimethylsulfamoyl)-2-methoxy-4-((8,8,8-trifluorooctyl)amino)benzoate,

[0211] 12.4 Methyl 5-(N,N-dimethylsulfamoyl)-2-hydroxy-4-((8,8,8-trifluorooctyl)amino)benzoate,

[0212] 12.5 Methyl 5-(N,N-dimethylsulfamoyl)-2-ethoxy-4-((8,8,8-trifluorooctyl)amino)benzoate,

[0213] 12.6 Methyl 2-(cyclopentyloxy)-5-(N,N-dimethylsulfamoyl)-4-((8,8,8-trifluorooctyl)amino)benzoate,

[0214] 12.7 5-(N,N-dimethylsulfamoyl)-2-ethoxy-4-((8,8,8-trifluorooctyl)amino)benzoic acid,

[0215] 12.8 2-(Cyclopentyloxy)-5-(N,N-dimethylsulfamoyl)-4-((8,8,8-trifluorooctyl)amino)benzoic acid,

[0216] 13.1 5-(N,N-dimethylsulfamoyl)-2-methoxy-4-((8,8,8-trifluorooctyl)amino)benzoic acid,

[0217] 14.3 3-Morpholinosulfonyl-4-((8,8,8-trifluorooctyl)amino)benzoic acid,

[0218] 14.4 3-((4,4-difluoropiperidin-1-yl)sulfonyl)-4-((8,8,8-trifluorooctyl)amino)benzoic acid,

[0219] 15.1 3-(Dimethylsulfamoyl)-4-(hept-6-enylamino)benzoic acid,

[0220] 15.2 Methyl 3-(N,N-dimethylsulfamoyl)-4-(hept-6-en-1-ylamino)benzoate,

[0221] 15.3 Methyl 4-((8-bromo-8,8-difluorooctyl)amino)-3-(N,N-dimethylsulfamoyl)benzoate,

[0222] 15.4 4-[(8-Bromo-8,8-difluoro-octyl)amino]-3-(dimethylsulfamoyl)benzoic acid,

[0223] 16.1 5-(Dimethylsulfamoyl)-2-isopropoxy-4-(8,8,8-trifluorooctylamino)benzoic acid,

[0224] 16.2 2-(Cyclohexyloxy)-5-(dimethylsulfamoyl)-4-(8,8,8-trifluorooctylamino)benzoic acid,

[0225] 16.3 5-(Dimethylsulfamoyl)-2-tetrahydropyran-4-yloxy-4-(8,8,8-trifluorooctylamino)benzoic acid,

[0226] 16.4 2-(Cyclobutyloxy)-5-(dimethylsulfamoyl)-4-(8,8,8-trifluorooctylamino)benzoic acid,

[0227] 16.5 5-(dimethylsulfamoyl)-2-(oxetan-3-yloxy)-4-(8,8,8-trifluorooctylamino)benzoic acid,

[0228] 16.6 5-(Dimethylsulfamoyl)-2-(4-piperidinyloxy)-4-(8,8,8-trifluorooctylamino)benzoic acid,

[0229] 16.7 5-(Dimethylsulfamoyl)-2-phenoxy-4-(8,8,8-trifluorooctylamino)benzoic acid.

[0230] Preferably, the compound of formula Ia is selected from the group consisting of:

[0231] 1.7 2-(Hexylamino)-5-nitrobenzenesulfonamide,

[0232] 1.17 2-(3,3-Dimethylbutylamino)-N,N-dimethyl-5-nitrobenzenesulfonamide,

[0233] 2.2 4-(Butylamino)-2-chloro-5-sulfamoyl-benzoic acid,

[0234] 2.6 4-(Butylamino)-3-sulfamoyl-benzoic acid,

[0235] 2.7 4-(Hexylamino)-3-sulfamoyl-benzoic acid,

[0236] 2.8 4-(Octylamino)-3-sulfamoyl-benzoic acid,

[0237] 2.9 4-(3,3-Dimethylbutylamino)-3-sulfamoyl-benzoic acid,

[0238] 3.6 4-(Butylamino)-3-(methylsulfamoyl)benzoic acid,

[0239] 3.7 4-(Hexylamino)-3-(methylsulfamoyl)benzoic acid,

[0240] 3.8 3-(Methylsulfamoyl)-4-(octylamino)benzoic acid,

[0241] 3.9 4-(3,3-dimethylbutylamino)-3-(methylsulfamoyl)benzoic acid,

[0242] 3.10 3-(Methylsulfamoyl)-4-(8,8,8-trifluorooctylamino)benzoic acid,

[0243] 3.11 4-(Butylamino)-3-(dimethylsulfamoyl)benzoic acid,

[0244] 3.12 3-(Dimethylsulfamoyl)-4-(hexylamino)benzoic acid,

[0245] 3.13 3-(Dimethylsulfamoyl)-4-(octylamino)benzoic acid,

[0246] 3.14 4-(3,3-Dimethylbutylamino)-3-(dimethylsulfamoyl)benzoic acid,

[0247] 3.17 3-(Dimethylsulfamoyl)-4-(8,8,8-trifluorooctylamino)benzoic acid,

[0248] 3.20 3-(Dimethylsulfamoyl)-4-(6-methoxyhexylamino)benzoic acid,

[0249] 3.21 3-(Cyclopentylsulfamoyl)-4-(8,8,8-trifluorooctylamino)benzoic acid,

[0250] 3.22 3-(Cyclohexylsulfamoyl)-4-(8,8,8-trifluorooctylamino)benzoic acid,

[0251] 5.5 3-Pyrrolidin-1-ylsulfonyl-4-(8,8,8-trifluorooctylamino)benzoic acid,

[0252] 5.6 3-(1-piperidinylsulfonyl)-4-(8,8,8-trifluorooctylamino)benzoic acid,

[0253] 5.7 3-Morpholinosulfonyl-4-(8,8,8-trifluorooctylamino)benzoic acid,

[0254] 13.1 5-(N,N-dimethylsulfamoyl)-2-methoxy-4-((8,8,8-trifluorooctyl)amino)benzoic acid,

[0255] 14.4 3-((4,4-difluoropiperidin-1-yl)sulfonyl)-4-((8,8,8-trifluorooctyl)amino)benzoic acid,

[0256] 15.1 3-(Dimethylsulfamoyl)-4-(hept-6-enylamino)benzoic acid.

[0257] In a further embodiment, the compound of Formula Ia is selected from the group consisting of:

[0258] 1.7 2-(Hexylamino)-5-nitrobenzenesulfonamide,

[0259] 1.15 2-(Hexylamino)-N,N-dimethyl-5-nitrobenzenesulfonamide,

[0260] 2.2 4-(Butylamino)-2-chloro-5-sulfamoyl-benzoic acid,

[0261] 2.6 4-(Butylamino)-3-sulfamoyl-benzoic acid,

[0262] 2.7 4-(Hexylamino)-3-sulfamoyl-benzoic acid,

[0263] 2.8 4-(Octylamino)-3-sulfamoyl-benzoic acid,

[0264] 3.8 3-(Methylsulfamoyl)-4-(octylamino)benzoic acid,

[0265] 3.13 3-(Dimethylsulfamoyl)-4-(octylamino)benzoic acid,

[0266] 3.14 4-(3,3-dimethylbutylamino)-3-(dimethylsulfamoyl)benzoic acid, and

[0267] 3.17 3-(Dimethylsulfamoyl)-4-(8,8,8-trifluorooctylamino)benzoic acid.

[0268] According to a second aspect of the present invention, there is provided a compound of formula Ib or a pharmaceutically acceptable salt or stereoisomeric form thereof, or an individual geometric isomer, enantiomer, diastereomer, tautomer, zwitterion and pharmaceutically acceptable salt thereof, for use as a medicament:

[0269]

[0270] in:

[0271] R1 and R2 are independently

[0272] ·hydrogen;

[0273] · Straight-chain or branched, unsubstituted or substituted C 1-10 an alkyl group, optionally containing one or more degrees of unsaturation;

[0274] · Straight-chain or branched, substituted or unsubstituted C 3-8 Cycloalkyl;

[0275] · Straight-chain or branched, substituted or unsubstituted C 4-10 cycloalkylalkyl;

[0276] ·C 3-8 heterocycloalkyl;

[0277] optionally substituted phenyl;

[0278] or R1 and R2 together with the nitrogen atom to which they are attached form a substituted or unsubstituted saturated heterocyclic ring;

[0279] R3 and R4 are independently

[0280] ·hydrogen;

[0281] Unsubstituted or substituted C 1-10 an alkyl group, optionally containing one or more degrees of unsaturation;

[0282] ·C 3-10 Cycloalkyl;

[0283] ·C 4-10 cycloalkylalkyl;

[0284] ·C 2-8 alkyl halide;

[0285] ·Straight chain or branched chain C 2-8 heteroalkyl, substituted or unsubstituted;

[0286] optionally substituted phenyl;

[0287] Provided that at least one of R3 and R4 is not hydrogen;

[0288] or R3 and R4, when taken together with the nitrogen atom to which they are attached, form a substituted or unsubstituted saturated heterocyclic ring;

[0289] R5 is

[0290] ·hydrogen;

[0291] ·halogen;

[0292] hydroxyl groups;

[0293] -OC 1-10 alkyl;

[0294] -OC 3-10 Cycloalkyl;

[0295] -OC 3-8 heterocycloalkyl;

[0296] ·C 1-10 Alkoxyalkyl;

[0297] ·C 3-10 Alkoxycycloalkyl;

[0298] optionally substituted phenoxy;

[0299] -NH2;

[0300] ·C 1-8 Alkylamines;

[0301] C2-C 16 dialkylamines;

[0302] ·aniline;

[0303] -SH;

[0304] ·C 1-8 Alkyl sulfides;

[0305] Thiophenol;

[0306] -NO2;

[0307] R6 is

[0308] Nitro;

[0309] Nitrile;

[0310] -CH2OH;

[0311] ·carboxylic acid;

[0312] ·C 1-4 alkyl esters;

[0313] ·C 2-8 heteroalkyl esters;

[0314] ·C3-6 Cycloalkyl esters;

[0315] Phenyl esters;

[0316] Carboxamides;

[0317] ·C 1-4 Alkylamides;

[0318] ·C 2-8 dialkylamides;

[0319] Cycloalkylamides;

[0320] Cyclic amides;

[0321] Tetrazole.

[0322] In a further embodiment, there is provided a compound of Formula Ic or a pharmaceutically acceptable salt or stereoisomeric form thereof, or an individual geometric isomer, enantiomer, diastereomer, tautomer, zwitterion and pharmaceutically acceptable salt thereof, for use as a medicament:

[0323]

[0324] in:

[0325] R1 and R2 are independently

[0326] ·hydrogen;

[0327] · Straight-chain or branched, unsubstituted or substituted C 1-10 an alkyl group, optionally containing one or more degrees of unsaturation;

[0328] · Straight-chain or branched, substituted or unsubstituted C 3-8 Cycloalkyl;

[0329] · Straight-chain or branched, substituted or unsubstituted C 4-10 cycloalkylalkyl;

[0330] optionally substituted phenyl;

[0331] or R1 and R2 together with the nitrogen atom to which they are attached form a substituted or unsubstituted saturated heterocyclic ring;

[0332] R3 and R4 are independently

[0333] ·hydrogen;

[0334] ·Substituted or unsubstituted C 1-10 an alkyl group, optionally containing one or more degrees of unsaturation;

[0335] ·C 3-10 Cycloalkyl;

[0336] ·C 4-10 cycloalkylalkyl;

[0337] ·C 2-8 alkyl halide;

[0338] ·Straight chain or branched chain C 2-8 heteroalkyl, substituted or unsubstituted;

[0339] optionally substituted phenyl;

[0340] Provided that at least one of R3 and R4 is not hydrogen;

[0341] or R3 and R4, when taken together with the nitrogen atom to which they are attached, form a substituted or unsubstituted saturated heterocyclic ring;

[0342] R5 is

[0343] ·hydrogen;

[0344] ·halogen;

[0345] hydroxyl groups;

[0346] ·C 1-10 Alkoxyalkyl;

[0347] ·C 3-10 Alkoxycycloalkyl;·optionally substituted phenoxy;·-NH2;

[0348] ·C 1-8 Alkylamines;

[0349] C2-C 16 dialkylamines;

[0350] ·aniline;

[0351] -SH;

[0352] ·C 1-8 Alkyl sulfides;

[0353] Thiophenol;

[0354] -NO2;

[0355] R6 is

[0356] Nitro;

[0357] Nitrile;

[0358] -CH2OH;

[0359] ·carboxylic acid;

[0360] ·C 1-4 alkyl esters;

[0361] ·C2-8 heteroalkyl esters;

[0362] ·C 3-6 Cycloalkyl esters;

[0363] Phenyl esters;

[0364] Carboxamides;

[0365] ·C 1-4 Alkylamides;

[0366] ·C 2-8 dialkylamides;

[0367] Cycloalkylamides;

[0368] Cyclic amides;

[0369] Tetrazole.

[0370] The compounds of Formula Ib and Ic are useful in the treatment or prevention of conditions in which there may be a component associated with depolarizing GABAergic transmission due to increased NKCC1 or decreased KCC2 expression levels or function.

[0371] In one embodiment of the present invention, there is provided a pharmaceutical composition comprising at least one compound of Formula Ib or Ic in a pharmaceutically acceptable carrier.

[0372] In a further embodiment, methods are provided for treating disorders associated with depolarizing GABAergic transmission due to increased NKCC1 or decreased KCC2 expression levels or function; for example, the methods can be performed by administering to a subject in need thereof a pharmaceutical composition containing a therapeutically effective amount of at least one compound of Formula Ib or Ic.

[0373] Advantageously, the method has been shown to have no diuretic side effects.

[0374] These compounds are useful in treating mammals, including humans.

[0375] In any given case, the actual amount of compound to be administered will be determined by the physician taking into account relevant circumstances, such as the severity of the condition, the age and weight of the patient, the patient's general physical condition, the cause of the condition, and the route of administration. In addition, the formulation can be designed to provide sustained release of the active compound over a given period of time, or to carefully control the amount of drug released at a given time during the course of treatment.

[0376] Given the chemical structure of the compounds of the present invention, suitable formulations can be prepared to allow effective amounts of the drug to pass through the blood-brain barrier; for example, nanoformulations can be prepared.

[0377] Because the severity of symptoms in individual subjects can vary widely and each drug has unique therapeutic properties, the precise administration and dosage employed for each subject is at the discretion of the practitioner.

[0378] 2-Aminobenzenesulfonamide derivatives have been shown to be potent inhibitors of the NKCC1 transporter, showing good percentage inhibition at 10 and 100 micromolar concentrations in cell-based assays. In addition, these compounds showed significant activity in a Down syndrome mouse model (Ts65Dn mice), rescuing hippocampal-dependent cognitive behaviors at a dose of 0.2 mg / kg. Notably, in C57Bl6N mice, Ts65Dn mice, and their wild-type littermates, in vivo treatment with these compounds had no statistically significant diuretic effect at 0.2 mg / kg compared to vehicle-treated animals. Additionally, in a drug-induced rodent model of autism, these compounds demonstrated significant efficacy in restoring sociability.

[0379] In a second aspect, the present invention relates to compounds of Formula Ib or Ic for use in treating diseases or conditions associated with depolarizing GABAergic transmission due to increased NKCC1 or decreased KCC2 (relative to physiological or desired) expression levels or function. In particular, the compounds described herein are useful in treating Down's syndrome, neuropathic pain, stroke, cerebral ischemia, cerebral edema, hydrocephalus, traumatic brain injury, depressive-like behavior induced by brain trauma, autism spectrum disorders (i.e., autism, fragile X, Rett syndrome, Asperger's and DiGeorge syndromes), epilepsy, convulsions, status epilepticus, West syndrome, glioma, glioblastoma, anaplastic astrocytoma, Parkinson's disease, Huntington's disease, schizophrenia, anxiety, tuberous sclerosis and related behavioral problems, Dravet syndrome.

[0380] The present invention can be used as a stand-alone therapeutic agent or in combination with other psychotropic drugs (including but not limited to fluoxetine, memantine, donepezil, DAPT), anti-inflammatory drugs (including but not limited to acetaminophen) and other COX inhibitors, antioxidants and psychoactive food supplements (including but not limited to melatonin, EGCG, resveratrol, omega-3, folinic acid, selenium, zinc, vitamins A, E and C). In addition, the present invention can be used in conjunction with early childhood education therapy.

[0381] In preferred embodiments, the compounds described herein are characterized by an amino substituent in an ortho position of the benzenesulfonamide scaffold, a carboxylic acid substituent of a unit of the benzenesulfonamide scaffold, the presence of an amino group and at least one substituent different from hydrogen, and the absence of aromatic substituents on the benzenesulfonamide scaffold.

[0382] Surprisingly, the compounds described herein demonstrated potent inhibition of NKCC1 compared to bumetanide.

[0383] As a further advantage, the compounds of the present invention have shown specific NKCC1 / NKCC2 selectivity, making them highly desirable.

[0384] Furthermore, the compounds according to the invention are characterized by the absence of a diuretic effect.

[0385] In a further advantage, the compounds of the present invention exhibit NKCC1 / NKCC2 selectivity without an associated diuretic effect.

[0386] In particular, compound 3.17 of the present invention, as disclosed below, showed the highest NKCC1 / NKCC2 selectivity.

[0387] Example

[0388] Example 1: Chemical synthesis and characterization

[0389] All commercial reagents and solvents were purchased from suppliers without further purification. Dry solvents were purchased from Sigma-Aldrich. Teledyne ISCO equipment ( Automated column chromatography purification was performed using mixtures of cyclohexane and ethyl acetate (EtOAc), cyclohexane and tert-butyl methyl ether (TBME), or dichloromethane (DCM) and methanol (MeOH) of increasing polarity. NMR experiments were run on a Bruker Avance III 400 system equipped with a BBI probe and a Z gradient (400.13 MHz for 1H and 100.62 MHz for 13C). Spectra were acquired at 300K using deuterated dimethyl sulfoxide (DMSO-d6) or deuterated chloroform (CDC13) as solvents. For 1H-NMR, data are reported as follows: chemical shift, multimodality (s=singlet, d=doublet, dd=doublet of doublet, t=triplet, q=quartet, m=multiplet), coupling constant (Hz), and integration. UPLC / MS analysis was performed on a Waters ACQUITY UPLC / MS system consisting of an SQD (single quadrupole detector) mass spectrometer equipped with an electrospray ionization interface and a photodiode array detector. The PDA range was 210-400 nm. Analysis was performed on an ACQUITY UPLC BEH C18 column (100 x 2.1 mm ID, 1.7 μm particle size) with a VanGuard BEH C18 precolumn (5 x 2.1 mm ID, 1.7 μm particle size). The mobile phases were 10 mM NH4OAc in H2O (A) at pH 5 adjusted with CH3COOH and 10 mM NH4OAc in CH3CN-H2O (95:5) at pH 5.0 (B). Three types of gradients were applied depending on the analysis: step 1 (5% to 100% mobile phase B over 3 minutes), step 2 (5% to 50% mobile phase B over 3 minutes), or step 3 (50% to 100% mobile phase B over 3 minutes). Electrospray ionization was applied in both positive and negative modes. Electrospray ionization was applied in both positive and negative modes. ESI was applied in both positive and negative modes. All test compounds showed a purity of ≥90% by NMR and UPLC / MS analysis.

[0390] Figure 5 Schemes and synthetic procedures for the preparation of some of the compounds of the present invention are described in .

[0391] synthesis:

[0392] 2-Chloro-5-nitro-benzenesulfonyl chloride (Compound 1.2, Scheme 1).

[0393] 1-Chloro-4-nitrobenzene 1.1 (500 mg, 3.14 mmol) was stirred in chlorosulfonic acid (1.05 ml, 15.71 mmol) at 120°C for 16 h. After completion of the reaction, the mixture was slowly poured into ice-cold water (30 ml) and extracted twice with DCM (2 x 30 ml). The combined organic layers were dried over NaSO and concentrated to dryness under reduced pressure to provide 374.1 mg (46% yield) of the title compound. Characterization: Rt = 2.14 min; MS (ESI) m / z: 253.7 [MH]-, calculated [MH]-: 254.9. 1H NMR (400 MHz, DMSO-d6) δ 8.61 (d, J = 2.9 Hz, 1H), 8.16 (dd, J = 8.7, 2.9 Hz, 1H), 7.70 (d, J = 8.6 Hz, 1H).

[0394] 2-Chloro-5-nitro-benzenesulfonamide (Compound 1.3, Scheme 1).

[0395] To 5ml tetrahydrofuran (THF) and 4ml 20% NH the ice-cold solution of the OH aqueous solution adds the compound 1.2 (374.1,1.47mmol) that is dissolved in THF, and at room temperature reaction mixture is stirred 1 hour.Then the crude reaction product is evaporated to dryness at low pressure, and residue is suspended in water (20ml) and extracted twice with EtOAc (2x20ml).Use Na sO dry the organic layer merged and be concentrated into dryness at low pressure.By flash chromatography on silica gel (cyclohexane / EtOAc, from 90: 10 to 70: 30), pure title compound (166.2g, yield 48%) is provided. Characterization: Rt = 1.42 min; MS (ESI) m / z: 235.3 [MH]-, calculated [MH]-: 236. 1H NMR (400 MHz, DMSO-d6) δ 8.68 (d, J = 2.7 Hz, 1H), 8.42 (dd, J = 8.7, 2.8 Hz, 1H), 7.98 (s, 2H), 7.96 (m, J = 8.7 Hz, 1H).

[0396] General Procedure C for the synthesis of sulfonamides 1.4-1.5 (Reaction C, Scheme 1).

[0397] To the ice-cold solution of suitable amine hydrochloride (1.0mmol) and triethylamine (2mmol) in DCM (1.0ml), add the compound 1.2 (1mmol) that is dissolved in DCM (1.5ml), and reaction mixture was at room temperature stirred 1 hour. The reaction crude product is diluted with DCM (20ml), and NH is washed with a saturated solution of Cl (20ml), and DCM (2x20ml) is used for water layer. Use Na sO dry the organic layer that merges, and be concentrated into dryness under low pressure. Finally, by flash chromatography on silica gel, pure title compound is obtained.

[0398] 2-Chloro-N-methyl-5-nitro-benzenesulfonamide (Compound 1.4, Scheme 1).

[0399] The title compound was synthesized according to the previously described General Procedure C using Intermediate 1.2 (347 mg, 1.46 mmol) and methylamine hydrochloride (100.7 mg, 1.46 mmol). Purification by flash chromatography on silica gel (cyclohexane / TBME 95:05) provided the pure title compound (204.9 mg, 56% yield). Characterization: Rt = 1.62 min; MS (ESI) m / z: 249.3 [MH]-. Calculated [MH]-: 250. 1H NMR (400 MHz, DMSO-d6) δ 8.61 (d, J = 2.7 Hz, 1H), 8.45 (dd, J = 8.7, 2.8 Hz, 1H), 8.11 (q, J = 4.4 Hz, 1H), 2.53 (d, J = 4.7 Hz, 3H).

[0400] 2-Chloro-N,N-dimethyl-5-nitro-benzenesulfonamide (Compound 1.5, Scheme 1)

[0401] The title compound was synthesized according to General Procedure C described previously using Intermediate 1.2 (190.3 mg, 0.8 mmol) and dimethylamine hydrochloride (163.7 mg, 1.60 mmol). Purification by flash chromatography on silica gel (cyclohexane / EtOAc 80:20) provided the pure title compound (156.32 mg, 74% yield). Characterization: Rt = 1.98 min; MS (ESI) m / z: 265.3 [MH]+. Calculated [MH]-: 264. 1H NMR (400 MHz, DMSO-d6) δ 8.59 (d, J = 2.7 Hz, 1H), 8.46 (dd, J = 8.7, 2.8 Hz, 1H), 8.01 (d, J = 8.7 Hz, 1H), 2.87 (s, 6H).

[0402] General Procedure D for the synthesis of compounds 1.6-1.17 (Reaction D, Scheme 1).

[0403] At 100 ℃, under argon atmosphere, a suspension of intermediate 1.3, 1.4 or 1.5 (1mmol) and appropriate amine (5mmol) in dry toluene (0.7ml) was stirred for 1 hour. After the reaction was completed, the mixture was evaporated to dryness under low pressure, and the residue was processed with water (10ml) and extracted with EtOAc (10ml). Use Na2SO4 dry organic layer and be concentrated to dryness under low pressure. Finally, pure title compound was obtained by flash chromatography on silica gel.

[0404] 2-(Butylamino)-5-nitro-benzenesulfonamide (Compound 1.6, Scheme 1).

[0405] The title compound was synthesized according to the previously described General Procedure D using Intermediate 1.3 (50 mg, 0.21 mmol) and butylamine (0.1 ml, 1.05 mmol). The compound was obtained pure (55.96 mg, 97% yield) without silica gel purification. Characterization: Rt = 2.03 min; MS (ESI) m / z: 274.4 [MH]+. Calculated [MH]-: 273.1; 1H NMR (400 MHz, DMSO-d6) δ 8.48 (d, J = 2.7 Hz, 1H), 8.19 (dd, J = 9.4, 2.7 Hz, 1H), 6.95 (d, J = 9.4 Hz, 1H), 3.35 (m, 2H), 1.65-1.55 (m, 2H), 1.44-1.32 (m, 2H), 0.92 (t, J = 7.3 Hz, 3H).

[0406] 2-(Hexylamino)-5-nitro-benzenesulfonamide (Compound 1.7, Scheme 1).

[0407] The title compound was synthesized using intermediate 1.3 (50 mg, 0.21 mmol) and hexylamine (0.14 ml, 1.05 mmol) according to the general procedure D described above. Purification by flash chromatography on silica gel (cyclohexane / EtOAc from 90:10 to 70:30) afforded the pure title compound (59.81 mg, 94% yield). Characterization: Rt = 2.34 min; MS (ESI) m / z: 302.5 [MH]+. Calculated [MH]-: 301.1; 1H NMR (400 MHz, DMSO-d6) δ 8.49 (d, J = 2.7 Hz, 1H), 8.19 (ddd, J = 9.4, 2.8, 0.5 Hz, 1H), 7.72 (s, 2H), 6.95 (d, J = 9.4 Hz, 1H), 6.85 (t, J = 5.6 Hz, 1H), 3.37-3.28 (m, 2H), 1.66-1.56 (m, 2H), 1.41-1.25 (m, 6H), 0.90-0.83 (m, 3H).

[0408] 5-Nitro-2-(octylamino)benzenesulfonamide (Compound 1.8, Scheme 1).

[0409] The title compound was synthesized using intermediate 1.3 (50 mg, 0.21 mmol) and octylamine (0.175 ml, 1.05 mmol) according to the general procedure D described above. Purification by flash chromatography on silica gel (cyclohexane / EtOAc 80:20) afforded the pure title compound (64.27 mg, 93% yield). Characterization: Rt = 2.61 min; MS (ESI) m / z: 330.5 [MH]+. Calculated [MH]-: 329.1; 1H NMR (400 MHz, DMSO-d6) δ 8.49 (d, J = 2.8 Hz, 1H), 8.20 (dd, J = 9.4, 2.8 Hz, 1H), 7.73 (s, 2H), 6.95 (d, J = 9.4 Hz, 1H), 6.86 (s, 1H), 3.34-3.29 (m, 2H), 1.62 (p, J = 7.2 Hz, 2H), 1.41-1.20 (m, 10H), 0.90-0.81 (m, 3H).

[0410] 2-(3,3-Dimethylbutylamino)-5-nitro-benzenesulfonamide (Compound 1.9, Scheme 1).

[0411] The title compound was synthesized using intermediate 1.3 (50 mg, 0.21 mmol) and 3,3-dimethylbutan-1-amine (0.148 ml, 1.05 mmol) according to the general procedure D described above. Purification by flash chromatography on silica gel (cyclohexane / EtOAc from 95:05 to 75:25) afforded the pure title compound (55.6 mg, 88% yield). Characterization: Rt = 2.29 min; MS (ESI) m / z: 265.3 [MH]+. Calculated [MH]-: 264; 1H NMR (400 MHz, DMSO-d6) δ 8.48 (d, J = 2.7 Hz, 1H), 8.21 (dd, J = 9.4, 2.8 Hz, 1H), 7.70 (s, 2H), 6.93 (d, J = 9.4 Hz, 1H), 6.78 (t, J = 4.7 Hz, 1H), 3.38-3.30 (m, 2H), 1.59-1.51 (m, 2H), 0.96 (s, 9H).

[0412] 2-(Butylamino)-N-methyl-5-nitro-benzenesulfonamide (Compound 1.10, Scheme 1).

[0413] The title compound was synthesized using intermediate 1.4 (40 mg, 0.16 mmol) and butylamine (80 μl, 0.79 mmol) according to the general procedure D described above. Purification by flash chromatography on silica gel (cyclohexane / EtOAc 80:20) afforded the pure title compound (38.65 mg, 84% yield). Characterization: Rt = 2.27 min; MS (ESI) m / z: 288.4 [MH]+. Calculated [MH]-: 287.1; 1H NMR (400 MHz, DMSO-d6) δ 8.40 (d, J = 2.8 Hz, 1H), 8.21 (dd, J = 9.4, 2.7 Hz, 1H), 7.89 (s, 1H), 6.98 (d, J = 9.4 Hz, 1H), 6.88 (t, J = 5.6 Hz, 1H), 3.38-3.33 (m, 2H), 2.44 (s, 3H), 1.66-1.54 (m, 2H), 1.43-1.32 (m, 2H), 0.92 (t, J = 7.4 Hz, 3H).

[0414] 2-(Hexylamino)-N-methyl-5-nitro-benzenesulfonamide (Compound 1.11, Scheme 1).

[0415] The title compound was synthesized using intermediate 1.4 (40 mg, 0.16 mmol) and hexylamine (0.1 ml, 0.79 mmol) according to the general procedure D described above. Purification by flash chromatography on silica gel (cyclohexane / EtOAc 80:20) provided the pure title compound (40.38 mg, 80% yield). Characterization: Rt = 2.56 min; MS (ESI) m / z: 316.4 [MH]+. Calculated [MH]-: 315.1; 1H NMR (400 MHz, DMSO-d6) δ 8.40 (d, J = 2.8 Hz, 1H), 8.21 (dd, J = 9.4, 2.8 Hz, 1H), 7.88 (s, 1H), 6.97 (d, J = 9.5 Hz, 1H), 6.92 (t, J = 5.6 Hz, 1H), 3.38-3.27 (m, 2H), 2.44 (s, 3H), 1.66-1.54 (m, 2H), 1.40-1.24 (m, 6H), 0.90-0.82 (m, 3H).

[0416] N-Methyl-5-nitro-2-(octylamino)benzenesulfonamide (Compound 1.12, Scheme 1).

[0417] The title compound was synthesized according to the general procedure D described above using intermediate 1.4 (40 mg, 0.16 mmol) and octylamine (0.13 ml, 0.79 mmol). Purification by flash chromatography on silica gel (cyclohexane / EtOAc 80:20) provided the pure title compound (39.56 mg, 72% yield). Characterization: Rt = 1.99 min; MS (ESI) m / z: 344.4 [MH]+. Calculated [MH]-: 343.1; 1H NMR (400 MHz, DMSO-d6) δ 8.41 (d, J = 2.8 Hz, 1H), 8.22 (dd, J = 9.4, 2.8 Hz, 1H), 7.89 (s, 1H), 6.98 (d, J = 9.4 Hz, 1H), 6.89 (t, J = 5.5 Hz, 1H), 3.36-3.30 (m, 2H), 2.45 (s, 3H), 1.65-1.56 (m, 2H), 1.40-1.20 (m, 1OH), 0.89-0.82 (m, 3H).

[0418] 2-(3,3-Dimethylbutylamino)-N-methyl-5-nitro-benzenesulfonamide (Compound 1.13, Scheme 1).

[0419] The title compound was synthesized using intermediate 1.4 (40 mg, 0.16 mmol) and 3,3-dimethylbutan-1-amine (0.11 ml, 0.79 mmol) according to the general procedure D described above. Purification by flash chromatography on silica gel (cyclohexane / EtOAc 80:20) provided the pure title compound (42.26 mg, 84% yield). Characterization: Rt = 2.15 min; MS (ESI) m / z: 316.4 [MH]+. Calculated [MH]-: 315.1; 1H NMR (400 MHz, DMSO-d6) δ 8.40 (d, J = 2.7 Hz, 1H), 8.23 ​​(dd, J = 9.3, 2.8 Hz, 1H), 6.96 (d, J = 9.4 Hz, 1H), 6.81 (t, J = 5.4 Hz, 1H), 3.36-3.30 (m, 2H), 2.43 (s, 3H), 1.57-1.51 (m, 2H), 0.96 (s, 9H).

[0420] 2-(Butylamino)-N,N-dimethyl-5-nitro-benzenesulfonamide (Compound 1.14, Scheme 1).

[0421] The title compound was synthesized using intermediate 1.5 (50 mg, 0.19 mmol) and butylamine (93 μl, 0.94 mmol) according to the general procedure D described above. Purification by flash chromatography on silica gel (cyclohexane / EtOAc 75:25) afforded the pure title compound (41.45 mg, 72% yield). Characterization: Rt = 2.47 min; MS (ESI) m / z: 302.4 [MH]+. Calculated [MH]-: 301.1; 1H NMR (400 MHz, DMSO-d6) δ 8.29 (d, J = 2.8 Hz, 1H), 8.25 (ddd, J = 9.4, 2.7, 0.6 Hz, 1H), 7.21 (t, J = 5.6 Hz, 1H), 7.03 (d, J = 9.5 Hz, 1H), 3.38-3.32 (m, 2H), 2.72 (s, 6H), 1.63-1.53 ​​(m, 2H), 1.42-1.32 (m, 2H), 0.93 (t, J = 7.3 Hz, 3H).

[0422] 2-(Hexylamino)-N,N-dimethyl-5-nitro-benzenesulfonamide (Compound 1.15, Scheme 1).

[0423] The title compound was synthesized using intermediate 1.5 (65 mg, 0.24 mmol) and hexylamine (0.16 ml, 1.21 mmol) according to the general procedure D described above. Purification by flash chromatography on silica gel (cyclohexane / EtOAc 80:20) afforded the pure title compound (68.42 mg, 87% yield). Characterization: Rt = 1.80 min; MS (ESI) m / z: 328.5 [MH]-. Calculated [MH]-: 329.1; 1H NMR (400 MHz, DMSO-d6) δ 8.28 (d, J = 2.7 Hz, 1H), 8.24 (ddd, J = 9.4, 2.8, 0.6 Hz, 1H), 7.21 (t, J = 5.6 Hz, 1H), 7.01 (d, J = 9.4 Hz, 1H), 3.36-3.30 (m, 2H), 2.71 (s, 6H), 1.62-1.53 ​​(m, 2H), 1.38-1.24 (m, 6H), 0.90-0.82 (m, 3H).

[0424] N,N-Dimethyl-5-nitro-2-(octylamino)benzenesulfonamide (Compound 1.16, Scheme 1).

[0425] The title compound was synthesized using intermediate 1.5 (50 mg, 0.19 mmol) and octylamine (0.15 ml, 0.94 mmol) according to the general procedure D described above. Purification by flash chromatography on silica gel (cyclohexane / EtOAc 85:15) afforded the pure title compound (57.52 mg, 85% yield). Characterization: Rt = 2.30 min; MS (ESI) m / z: 358.4 [MH]+. Calculated [MH]-: 357.2; 1H NMR (400 MHz, DMSO-d6) δ 8.28 (d, J = 2.8 Hz, 1H), 8.23 ​​(ddd, J = 9.4, 2.8, 0.6 Hz, 1H), 7.20 (t, J = 5.6 Hz, 1H), 7.01 (d, J = 9.5 Hz, 1H), 3.38-3.31 (m, 2H), 2.71 (s, 6H), 1.62-1.53 ​​(m, 2H), 1.37-1.20 (m, 1OH), 0.87-0.82 (m, 3H).

[0426] 2-(3,3-Dimethylbutylamino)-N,N-dimethyl-5-nitro-benzenesulfonamide (Compound 1.17, Scheme 1).

[0427] The title compound was synthesized using intermediate 1.5 (50 mg, 0.19 mmol) and 3,3-dimethylbutan-1-amine (0.13 ml, 0.94 mmol) according to the general procedure D described above. Purification by flash chromatography on silica gel (cyclohexane / EtOAc 85:15) afforded the pure title compound (51.11 mg, 82% yield). Characterization: Rt = 2.70 min; MS (ESI) m / z: 330.4 [MH]+. Calculated [MH]-: 329.1; 1H NMR (400 MHz, DMSO-d6) δ 8.28 (d, J = 2.7 Hz, 1H), 8.25 (ddd, J = 9.3, 2.8, 0.6 Hz, 1H), 7.16 (t, J = 5.6 Hz, 1H), 6.98 (d, J = 9.3 Hz, 1H), 3.38-3.32 (m, 2H), 2.71 (s, 6H), 1.52-1.47 (m, 2H), 0.95 (s, 9H).

[0428] General Procedure E for the Synthesis of Compounds 2.2-2.5 (Scheme 2) .

[0429] At 100 ℃, under argon atmosphere, a suspension of commercial 2-chloro-4-fluoro-5-sulfamoyl-benzoic acid 2.1 (1mmol) and the appropriate amine (5mmol) in dry toluene (0.7ml) was stirred for 1 hour. After the reaction was complete, the mixture was evaporated to dryness under low pressure, and the residue was treated with saturated NH4Cl aqueous solution (15ml) and extracted with EtOAc (15ml). Use Na2SO4 to dry the combined organic layers and concentrate to dryness under low pressure. Pulverize in cyclohexane to finally obtain the pure title compound.

[0430] 4-(Butylamino)-2-chloro-5-sulfamoyl-benzoic acid (Compound 2.2, Scheme 2).

[0431] The title compound was synthesized using intermediate 2.1 (70 mg, 0.26 mmol) and butylamine (0.13 ml, 1.32 mmol) according to the general procedure E described above. Trituration with cyclohexane (1 ml) afforded the pure title compound (40.84 mg, 51% yield). Characterization: Rt = 1.52 min; MS (ESI) m / z: 305.3 [MH]-. Calculated [MH]-: 306.04; 1H NMR (400 MHz, DMSO-d6) δ 12.80 (bs, 1H), 8.26 (s, 1H), 7.57 (s, 2H), 6.84 (s, 1H), 6.39 (t, J = 5.3 Hz, 1H), 3.31-3.21 (m, 2H), 1.64-1.53 ​​(m, 2H), 1.44-1.33 (m, 2H), 0.93 (t, J = 7.3 Hz, 3H).

[0432] 2-Chloro-4-(hexylamino)-5-sulfamoyl-benzoic acid (Compound 2.3, Scheme 2).

[0433] The title compound was synthesized using intermediate 2.1 (50 mg, 0.19 mmol) and hexylamine (0.12 ml, 0.95 mmol) according to the general procedure E described above. Trituration with cyclohexane (1 ml) afforded the pure title compound (52.82 mg, 83% yield). Characterization: Rt = 1.78 min; MS (ESI) m / z: 333.4 [MH]-. Calculated [MH]-: 334.1; 1H NMR (400 MHz, DMSO-d6) δ 12.77 (bs, 1H), 8.25 (s, 1H), 7.55 (s, 2H), 6.83 (s, 1H), 6.39 (t, J = 5.4 Hz, 1H), 3.27-3.20 (m, 2H), 1.59 (p, J = 7.1 Hz, 2H), 1.41-1.24 (m, 6H), 0.90-0.84 (m, 3H).

[0434] 2-Chloro-4-(octylamino)-5-sulfamoyl-benzoic acid (Compound 2.4, Scheme 2).

[0435] The title compound was synthesized using intermediate 2.1 (50 mg, 0.19 mmol) and octylamine (0.16 ml, 0.95 mmol) according to the general procedure E described above. Trituration with cyclohexane (1 ml) afforded the pure title compound (48.89 mg, 71% yield). Characterization: Rt = 2.01 min; MS (ESI) m / z: 361.4 [MH]-. Calculated [MH]-: 362.1; 1H NMR (400 MHz, DMSO-d6) δ 12.78 (bs, 1H), 8.26 (s, 1H), 7.56 (s, 2H), 6.84 (s, 1H), 6.40 (t, J 5.3 Hz, 1H), 3.28 3.21 (m, 2H), 1.65-1.55 (m, 2H), 1.41-1.20 (m, 10H), 0.90-0.83 (m, 3H).

[0436] 2-Chloro-4-(3,3-dimethylbutylamino)-5-sulfamoyl-benzoic acid (Compound 2.5, Scheme 2).

[0437] The title compound was synthesized according to General Procedure E described previously using Intermediate 2.1 (50 mg, 0.19 mmol) and 3,3-dimethylbutan-1-amine (0.13 mL, 0.95 mmol). Trituration with cyclohexane (1 mL) afforded the pure title compound (52.82 mg, 83% yield). Characterization: Rt = 1.66 min; MS (ESI) m / z: 333.4 [MH]-. Calculated [MH]-: 334.1; 1H NMR (400 MHz, DMSO-d6) δ 8.25 (s, 1H), 7.54 (s, 2H), 6.83 (s, 1H), 6.29 (t, J = 5.1 Hz, 1H), 3.27-3.20 (m, 2H), 1.56-1.50 (m, 2H), 0.96 (s, 9H).

[0438] General Procedure F for the synthesis of compounds 2.6-2.9 (Reaction F, Scheme 2).

[0439] Under Ar atmosphere, to the suspension of suitable 4-amino-2-chloro-5-sulfamoyl-benzoic acid intermediate 2.2-2.5 (1mmol) and palladium hydroxide on carbon (20wt.%) in dry methanol (20ml), add ammonium formate (4mmol), and reaction mixture was stirred 1 hour at reflux temperature. After reaction was completed, crude product was filtered by diatomite thick sheet, and filtrate was concentrated to dryness under low pressure. Dry resistates was diluted in EtOAc (10ml) and washed with saturated NH4Cl solution (10ml). Use Na2SO4 dry organic layer and be concentrated to dryness under low pressure. Pulverize in cyclohexane, finally obtain pure title compound.

[0440] 4-(Butylamino)-3-sulfamoyl-benzoic acid (Compound 2.6, Scheme 2).

[0441] The title compound was synthesized using intermediate 2.2 (30 mg, 0.1 mmol) according to the general procedure F described above. Trituration with cyclohexane (1 ml) gave the pure title compound (11.71 mg, 43% yield). Characterization: Rt = 1.53 min; MS (ESI) m / z: 273.4 [MH]+. Calculated [MH]-: 272.1; 1H NMR (400 MHz, DMSO-d6) δ 8.23 ​​(d, J = 2.1 Hz, 1H), 7.87 (dd, J = 8.8, 2.2 Hz, 1H), 7.46 (s, 2H), 6.83 (d, J = 8.9 Hz, 1H), 6.37 (t, J = 5.4 Hz, 1H), 3.28-3.21 (m, 2H), 1.64-1.55 (m, 2H), 1.44-1.34 (m, 2H), 0.92 (t, J = 7.3 Hz, 3H).

[0442] 4-(Hexylamino)-3-sulfamoyl-benzoic acid (Compound 2.7, Scheme 2).

[0443] The title compound was synthesized using intermediate 2.3 (30.7 mg, 0.09 mmol) according to the general procedure F described above. Trituration with cyclohexane (1 ml) gave the pure title compound (11.71 mg, 43% yield). Characterization: Rt = 1.81 min; MS (ESI) m / z: 301.4 [MH]+. Calculated [MH]-: 300.1; 1H NMR (400 MHz, DMSO-d6) δ 12.45 (bs, 1H), 8.23 ​​(d, J = 2.1 Hz, 1H), 7.87 (dd, J = 8.8, 2.2 Hz, 1H), 7.46 (s, 2H), 6.82 (d, J = 8.9 Hz, 1H), 6.38 (t, J = 5.4 Hz, 1H), 3.27-3.20 (m, 2H), 1.60 (h, J = 6.6 Hz, 2H), 1.42-1.25 (m, 8H), 0.92-0.80 (m, 3H).

[0444] 4-(Octylamino)-3-sulfamoyl-benzoic acid (Compound 2.8, Scheme 2).

[0445] The title compound was synthesized using intermediate 2.4 (35.7 mg, 0.1 mmol) according to the general procedure F described above. Trituration with cyclohexane (1 ml) gave the pure title compound (9.68 mg, 36% yield). Characterization: Rt = 2.16 min; MS (ESI) m / z: 329.4 [MH]+. Calculated [MH]-: 328.1; 1H NMR (400 MHz, DMSO-d6) δ 12.43 (bs, 1H), 8.23 ​​(d, J = 2.1 Hz, 1H), 7.86 (dd, J = 8.7, 2.1 Hz, 1H), 7.46 (s, 2H), 6.82 (d, J = 8.9 Hz, 1H), 6.38 (t, J = 5.3 Hz, 1H), 3.27-3.19 (m, 2H), 1.65-1.56 (m, 2H), 1.42-1.15 (m, 12H), 0.92-0.80 (m, 3H).

[0446] 4-(3,3-Dimethylbutylamino)-3-sulfamoyl-benzoic acid (Compound 2.9, Scheme 2).

[0447] The title compound was synthesized using Intermediate 2.5 (29.6 mg, 0.09 mmol) according to General Procedure F described above. Trituration with cyclohexane (1 ml) afforded the pure title compound (15.13 mg, 56% yield). Characterization: Rt = 1.80 min; MS (ESI) m / z: 301.4 [MH]+. Calculated [MH]-: 300.1; 1H NMR (400 MHz, DMSO-d6) δ 12.48 (bs, 1H), 8.24 (d, J = 2.1 Hz, 1H), 7.89 (dd, J = 8.8, 2.1 Hz, 1H), 7.46 (s, 2H), 6.83 (d, J = 8.9 Hz, 1H), 3.28-3.21 (m, 2H), 1.59-1.52 (m, 2H), 0.97 (s, 9H).

[0448] General Procedure G for the synthesis of compounds 3.2-3.3 (Reaction G, Scheme 3).

[0449] 4-Fluoro-3-chlorosulfonyl-benzoic acid 3.1 (1 mmol) dissolved in 1.5 mL of THF was added dropwise to 3 mL of an ice-cold 2 M THF solution of the appropriate solvent and stirred at room temperature for 1 h. After completion of the reaction, the reaction mixture was evaporated to dryness, and the residue was treated with water and HCl. The precipitated product was filtered and rinsed with water to obtain the pure title compound.

[0450] 4-Fluoro-3-(methylsulfamoyl)benzoic acid (Compound 3.2, Scheme 3).

[0451] The title compound was synthesized according to the previously described General Procedure G using intermediate 3.1 (500 mg, 2.07 mmol) and 2M methylamine in THF (2.07 ml, 4.15 mmol). The described workup afforded the pure title compound (313.8 mg, 64% yield). Characterization: Rt = 1.26 min; MS (ESI) m / z: 232.3 [MH]-. Calculated [MH]-: 233.02. 1H NMR (400 MHz, DMSO-d6) δ 8.30 (dd, J = 7.0, 2.2 Hz, 1H), 8.25-8.19 (m, 1H), 7.89 (q, J = 4.8 Hz, 1H), 7.62-7.54 (m, 1H), 2.52 (d, J = 4.8 Hz, 3H).

[0452] 3-(Dimethylsulfamoyl)-4-fluoro-benzoic acid (Compound 3.3, Scheme 3).

[0453] The title compound was synthesized according to the previously described General Procedure G using intermediate 3.1 (1 g, 4.15 mmol) and a 2M solution of dimethylamine in THF (4.15 ml, 8.30 mmol). The described workup afforded the pure title compound (749 mg, 73% yield). Characterization: Rt = 1.11 min; MS (ESI) m / z: 246.3 [MH]-. Calculated [MH]-: 247.03. 1H NMR (400 MHz, DMSO-d6) δ 8.29-8.24 (m, 2H), 7.67-7.58 (m, 1H), 2.75 (d, J = 1.9 Hz, 6H).

[0454] 3-(Cyclopentylsulfamoyl)-4-fluoro-benzoic acid (Compound 3.4, Scheme 3).

[0455] The title compound was synthesized according to the previously described General Procedure G using intermediate 3.1 (250 mg, 1.04 mmol) and cyclopentylamine (0.21 ml, 2.07 mmol) in THF (8.5 ml). The described workup afforded the pure title compound (261.4 mg, 88% yield). Characterization: Rt = 1.25 min; MS (ESI) m / z: 286.4 [MH]-. Calculated MH]-: 287.06. 1H NMR (400 MHz, DMSO-d6) δ 8.33 (dd, J = 7.1, 2.3 Hz, 1H), 8.21 (ddd, J = 8.6, 4.7, 2.3 Hz, 1H), 8.12 (d, J = 7.6 Hz, 1H), 7.56 (dd, J = 10.0, 8.6 Hz, 1H), 3.58-3.48 (m, 1H), 1.68-1.48 (m, 4H), 1.45-1.28 (m, 4H).

[0456] 3-(Cyclohexylsulfamoyl)-4-fluoro-benzoic acid (Compound 3.5, Scheme 3).

[0457] The title compound was synthesized according to the previously described General Procedure G using intermediate 3.1 (250 mg, 1.04 mmol) and cyclohexylamine (0.24 ml, 2.07 mmol) in THF (8.5 ml). The described workup and trituration with a 9:1 cyclohexane / ethyl acetate mixture (2 ml) afforded the pure title compound (185.6 mg, 59% yield). Characterization: Rt = 1.37 min; MS (ESI) m / z: 286.4 [MH]-. Calculated [MH]-: 287.06. 1H NMR (400 MHz, DMSO-d6) δ8.33 (dd, J=7.1, 2.3 Hz, 1H), 8.21 (ddd, J=8.6, 4.7, 2.3 Hz, 1H), 8.12 (d, J=7.6 Hz, 1H), 7.56 (dd, J=10.0, 8.6 Hz, 1H), 3.58-3.48 (m, 1H), 1.68-1.48 (m, 4H), 1.45-1.28 (m, 4H).

[0458] General Procedure H for the synthesis of compounds 3.6-3.22, 5.5-5.7, 6.3, 7.4 (Reaction H, Schemes 3, 5, 6, 7).

[0459] At 100 ° C under an argon atmosphere, a suspension of the appropriate intermediate (1 mmol) and the appropriate amine (2 mmol) in dry 1,4-dioxane (3 ml) was stirred for 4 hours. After the reaction was complete, the mixture was evaporated to dryness under low pressure, and the residue was treated with a saturated NH4Cl aqueous solution (15 ml) and extracted twice with EtOAc (2x15 ml). The combined organic layers were dried using Na2SO4 and concentrated to dryness under low pressure. Pulverized in cyclohexane to finally obtain the pure title compound.

[0460] 4-(Butylamino)-3-(methylsulfamoyl)benzoic acid (Compound 3.6, Scheme 3).

[0461] The title compound was synthesized according to the general procedure described above using intermediate 3.2 (50 mg, 0.21 mmol) and butylamine (42 μl, 0.42 mmol) in dry 1,4-dioxane (0.7 ml). Trituration with cyclohexane (1 ml) afforded the pure title compound (47.10 mg, 78% yield). Characterization: Rt = 1.66 min; MS (ESI) m / z: 285.4 [MH]-. Calculated [MH]-: 286.1. 1H NMR (400 MHz, DMSO-d6) δ8.15 (d, J = 2.1 Hz, 1H), 7.90 (dd, J = 8.8, 2.1 Hz, 1H), 7.66 (s, 1H), 6.86 (d, J = 8.9 Hz, 1H), 6.44 (t, J = 5.4 Hz, 1H), 3.24 (q, J = 6.6 Hz, 2H), 2.39 (s, 3H), 1.58 (p, J = 7.2 Hz, 2H), 1.43-1.32 (m, 2H), 0.92 (t, J = 7.3 Hz, 3H).

[0462] 4-(Hexylamino)-3-(methylsulfamoyl)benzoic acid (Compound 3.7, Scheme 3).

[0463] The title compound was synthesized according to the general procedure described above using intermediate 3.2 (50 mg, 0.21 mmol) and hexylamine (57 μl, 0.42 mmol) in dry 1,4-dioxane (0.7 ml). Trituration with cyclohexane (1 ml) afforded the pure title compound (51.69 mg, 78% yield). Characterization: Rt = 2.00 min; MS (ESI) m / z: 313.4 [MH]-. Calculated [MH]-: 314.1. 1H NMR (400 MHz, DMSO-d6) δ 12.53 (bs, 1H), 8.15 (d, J = 2.1 Hz, 1H), 7.90 (dd, J = 8.8, 2.1 Hz, 1H), 7.63 (q, J = 5.0 Hz, 1H), 6.86 (d, J = 8.9 Hz, 1H), 6.44 (t, J = 5.3 Hz, 1H), 3.23 (q, J = 6.6 Hz, 2H), 1.60 (p, J = 7.1 Hz, 2H), 1.40-1.25 (m, 6H), 0.90-0.83 (m, 3H).

[0464] 3-(Methylsulfamoyl)-4-(octylamino)benzoic acid (Compound 3.8, Scheme 3).

[0465] The title compound was synthesized according to the general procedure described above using intermediate 3.2 (50 mg, 0.21 mmol) and octylamine (71 μl, 0.42 mmol) in dry 1,4-dioxane (0.7 ml). Trituration with cyclohexane (1 ml) afforded the pure title compound (69.51 mg, 97% yield). Characterization: Rt = 2.28 min; MS (ESI) m / z: 341.4 [MH]-. Calculated [MH]-: 342.2. 1H NMR (400 MHz, DMSO-d6) δ8.15 (d, J = 2.1 Hz, 1H), 7.89 (dd, J = 8.8, 2.1 Hz, 1H), 6.86 (d, J = 8.9 Hz, 1H), 6.44 (t, J = 5.4 Hz, 1H), 3.23 (q, J = 6.6 Hz, 2H), 2.38 (s, 3H), 1.59 (p, J = 7.1 Hz, 2H), 1.40-1.20 (m, 9H), 0.89-0.82 (m, 3H).

[0466] 4-(3,3-Dimethylbutylamino)-3-(methylsulfamoyl)benzoic acid (Compound 3.9, Scheme 3).

[0467] The title compound was synthesized according to the general procedure described above using intermediate 3.2 (50 mg, 0.21 mmol) and 3,3-dimethylbutan-1-amine (60 μl, 0.42 mmol) in dry 1,4-dioxane (0.7 ml). Trituration with cyclohexane (1 ml) afforded the pure title compound (50.56 mg, 84% yield). Characterization: Rt = 1.93 min; MS (ESI) m / z: 313.4 [MH]-. Calculated [MH]-: 314.1. 1H NMR (400 MHz, DMSO-d6) δ 12.52 (s, 1H), 8.15 (d, J = 2.1 Hz, 1H), 7.91 (dd, J = 8.8, 2.1 Hz, 1H), 7.62 (q, J = 5.0 Hz, 1H), 6.86 (d, J = 8.9 Hz, 1H), 6.35 (t, J = 5.2 Hz, 1H), 3.27-3.20 (m, 2H), 2.38 (d, J = 5.0 Hz, 3H), 1.57-1.50 (m, 2H), 0.96 (s, 9H).

[0468] 3-(Methylsulfamoyl)-4-(8,8,8-trifluorooctylamino)benzoic acid (Compound 3.10, Scheme 3).

[0469] The title compound was synthesized in dry 1,4-dioxane (1.4 ml) using Intermediate 3.2 (100 mg, 0.42 mmol) and Intermediate 4.5 (86.4 mg, 0.47 mmol) according to the general procedure described above. Trituration with cyclohexane (2 ml) afforded the pure title compound (111.5 mg, 67% yield). Characterization: Rt = 2.11 min; MS (ESI) m / z: 395.2 [MH]-. Calculated [MH]-: 396.1. 1H NMR (400 MHz, DMSO-d6) δ8.15 (d, J = 2.1 Hz, 1H), 7.90 (dd, J = 8.8, 2.1 Hz, 1H), 7.63 (q, J = 5.0 Hz, 1H), 6.86 (d, J = 8.9 Hz, 1H), 6.44 (t, J = 5.4 Hz, 1H), 3.24 (q, J = 6.7 Hz, 2H), 2.39 (d, J = 4.8 Hz, 3H), 2.28-2.15 (m, 2H), 1.64-1.55 (m, 2H), 1.51-1.42 (m, 2H), 1.39-1.30 (m, 6H).

[0470] 4-(Butylamino)-3-(dimethylsulfamoyl)benzoic acid (Compound 3.11, Scheme 3).

[0471] The title compound was synthesized according to the general procedure described above using intermediate 3.3 (50 mg, 0.20 mmol) and butylamine (40 μl, 0.40 mmol) in dry 1,4-dioxane (0.7 ml). Trituration with cyclohexane (1 ml) afforded the pure title compound (41.45 mg, 69% yield). Characterization: Rt = 1.90 min; MS (ESI) m / z: 299.4 [MH]-. Calculated [MH]-: 300.1. 1H NMR (400 MHz, DMSO-d6) δ 12.62 (s, 1H), 8.05 (d, J = 2.1 Hz, 1H), 7.93 (dd, J 8.9, 2.1 Hz, 1H), 6.91 (d, J 9.0 Hz, 1H), 6.74 (t, J = 5.4 Hz, 1H), 3.29-3.19 (m, 2H), 2.66 (s, 6H), 1.61-1.52 (m, 2H), 1.42-1.31 (m, 2H), 0.92 (t, J = 7.3 Hz, 3H).

[0472] 3-(Dimethylsulfamoyl)-4-(hexylamino)benzoic acid (Compound 3.12, Scheme 3).

[0473] The title compound was synthesized according to the general procedure described above using intermediate 3.3 (50 mg, 0.20 mmol) and hexylamine (53 μl, 0.40 mmol) in dry 1,4-dioxane (0.7 ml). Trituration with cyclohexane (1 ml) afforded the pure title compound (53.20 mg, 81% yield). Characterization: Rt = 2.17 min; MS (ESI) m / z: 327.4 [MH]-. Calculated [MH]-: 328.1. 1H NMR (400 MHz, DMSO-d6) δ 12.63 (s, 1H), 8.04 (d, J = 2.1 Hz, 1H), 7.93 (dd, J = 8.8, 2.1 Hz, 1H), 6.90 (d, J = 9.0 Hz, 1H), 6.74 (t, J = 5.4 Hz, 1H), 3.28-3.18 (m, 2H), 2.65 (s, 6H), 1.57 (p, J = 7.0 Hz, 2H), 1.39-1.24 (m, 6H), 0.89-0.84 (m, 3H).

[0474] 3-(Dimethylsulfamoyl)-4-(octylamino)benzoic acid (Compound 3.13, Scheme 3).

[0475] The title compound was synthesized according to the general procedure described above using intermediate 3.3 (50 mg, 0.20 mmol) and octylamine (67 μl, 0.40 mmol) in dry 1,4-dioxane (0.7 ml). Trituration with cyclohexane (1 ml) afforded the pure title compound (59.9 mg, 84% yield). Characterization: Rt 2.44 min; MS (ESI) m / z: 355.4 [MH]-. Calculated [MH]-: 356.2. 1H NMR (400 MHz, DMSO-d6) δ 12.62 (s, 1H), 8.04 (d, J = 2.1 Hz, 1H), 7.93 (dd, J = 8.9, 2.1 Hz, 1H), 6.91 (d, J = 9.0 Hz, 1H), 6.75 (t, J = 5.4 Hz, 1H), 3.23 (q, J = 6.6 Hz, 2H), 2.65 (s, 6H), 1.57 (p, J = 6.9 Hz, 2H), 1.39-1.19 (m, 10H), 0.90-0.80 (m, 3H).

[0476] 4-(3,3-Dimethylbutylamino)-3-(dimethylsulfamoyl)benzoic acid (Compound 3.14, Scheme 3).

[0477] The title compound was synthesized according to the general procedure described above using intermediate 3.3 (50 mg, 0.20 mmol) and 3,3-dimethylbutan-1-amine (57 μl, 0.40 mmol) in dry 1,4-dioxane (0.7 ml). Trituration with cyclohexane (1 ml) afforded the pure title compound (42 mg, 63% yield). Characterization: Rt = 2.13 min; MS (ESI) m / z: 327.4 [MH]-. Calculated [MH]-: 328.1. 1H NMR (400 MHz, DMSO-d6) δ 12.63 (s, 1H), 8.05 (d, J = 2.0 Hz, 1H), 7.95 (dd, J = 8.9, 2.1 Hz, 1H), 6.90 (d, J = 8.9 Hz, 1H), 6.69 (t, J = 5.3 Hz, 1H), 3.29-3.22 (m, 2H), 2.66 (s, 6H), 1.54-1.46 (m, 2H), 0.96 (s, 9H).

[0478] 3-(Dimethylsulfamoyl)-4-(4,4,4-trifluorobutylamino)benzoic acid (Compound 3.15, Scheme 3).

[0479] The title compound was synthesized according to the general procedure described above using intermediate 3.3 (50 mg, 0.20 mmol) and 4,4,4-trifluorobutylamine (48 μl, 0.40 mmol) in dry 1,4-dioxane (0.7 ml). Trituration with cyclohexane (1 ml) afforded the pure title compound (40.13 mg, 57% yield). Characterization: Rt = 1.78 min; MS (ESI) m / z: 353.4 [MH]-. Calculated [MH]-: 354.1. 1H NMR (400 MHz, DMSO-d6) δ 12.64 (bs, 1H), 8.07 (d, J = 2.1 Hz, 1H), 7.95 (dd, J = 8.8, 2.1 Hz, 1H), 6.98 (d, J = 9.0 Hz, 1H), 6.88 (t, J = 5.9 Hz, 1H), 3.38 (q, J = 6.8 Hz, 2H), 2.67 (s, 6H), 2.40-2.25 (m, 2H), 1.83-1.73 (m, 2H).

[0480] 3-(Dimethylsulfamoyl)-4-(6,6,6-trifluorohexylamino)benzoic acid (Compound 3.16, Scheme 3).

[0481] The title compound was synthesized according to the general procedure described above using intermediate 3.3 (50 mg, 0.20 mmol) and 6,6,6-trifluorohexylamine (60 μl, 0.40 mmol) in dry 1,4-dioxane (0.7 ml). Trituration with cyclohexane (1 ml) afforded the pure title compound (57.32 mg, 75% yield). Characterization: Rt = 2.02 min; MS (ESI) m / z: 381.4 [MH]-. Calculated [MH]-: 382.1. 1H NMR (400 MHz, DMSO-d6) δ 12.64 (bs, 1H), 8.05 (d, J = 2.1 Hz, 1H), 7.94 (dd, J = 8.8, 2.1 Hz, 1H), 6.93 (d, J = 9.0 Hz, 1H), 6.77 (t, J = 5.4 Hz, 1H), 3.26 (q, J = 6.8 Hz, 2H), 2.66 (s, 6H), 2.32-2.18 (m, 2H), 1.62 (p, J = 7.4 Hz, 3H), 1.58-1.48 (m, 2H), 1.47-1.37 (m, 2H).

[0482] 3-(Dimethylsulfamoyl)-4-(8,8,8-trifluorooctylamino)benzoic acid (Compound 3.17, Scheme 3).

[0483] The title compound was synthesized according to the general procedure described above using intermediate 3.3 (50 mg, 0.20 mmol) and intermediate 4.5 (89 mg, 0.40 mmol) in dry 1,4-dioxane (0.7 ml). Trituration with cyclohexane (1 ml) afforded the pure title compound (44.34 mg, 54% yield). Characterization: Rt = 2.28 min; MS (ESI) m / z: 409.4 [MH]-. Calculated [MH]-: 410.1. 1H NMR (400 MHz, DMSO-d6) δ 12.62 (s, 1H), 8.05 (d, J = 2.1 Hz, 1H), 7.93 (dd, J = 8.8, 2.1 Hz, 1H), 6.91 (d, J = 9.0 Hz, 1H), 6.75 (t, J = 5.4 Hz, 1H), 3.24 (q, J = 6.6 Hz, 2H), 2.29-2.14 (m, 2H), 1.64-1.52 (m, 2H), 1.52-1.39 (m, 2H), 1.40-1.25 (m, 6H).

[0484] 3-(Dimethylsulfamoyl)-4-(2-methoxyethylamino)benzoic acid (Compound 3.18, Scheme 3).

[0485] The title compound was synthesized according to the general procedure described above using intermediate 3.3 (50 mg, 0.20 mmol) and 2-methoxyethylamine (36 μl, 0.40 mmol) in dry 1,4-dioxane (0.7 ml). Trituration with cyclohexane (1 ml) afforded the pure title compound (53.96 mg, 89% yield). Characterization: Rt = 1.40 min; MS (ESI) m / z: 301.4 [MH]-. Calculated [MH]-: 302.1. 1H NMR (400 MHz, DMSO-d6) δ8.05 (d, J = 2.1 Hz, 1H), 7.93 (dd, J = 8.8, 2.1 Hz, 1H), 6.95 (d, J = 9.0 Hz, 1H), 6.89 (t, J = 5.3 Hz, 1H), 3.55 (t, J = 5.2 Hz, 2H), 3.40 (q, J = 5.3 Hz, 2H), 3.29 (s, 3H), 2.65 (s, 6H).

[0486] 3-(Dimethylsulfamoyl)-4-(4-methoxybutylamino)benzoic acid (Compound 3.19, Scheme 3).

[0487] The title compound was synthesized according to the general procedure described above using intermediate 3.3 (50 mg, 0.20 mmol) and 4-methoxybutan-1-amine (51 μl, 0.40 mmol) in dry 1,4-dioxane (0.7 ml). Trituration with cyclohexane (1 ml) afforded the pure title compound (56.08 mg, 85% yield). Characterization: Rt = 1.59 min; MS (ESI) m / z: 329.4 [MH]-. Calculated [MH]-: 330.1. 1H NMR (400 MHz, DMSO-d6) δ 12.63 (s, 1H), 8.05 (d, J = 2.1 Hz, 1H), 7.93 (dd, J = 8.8, 2.1 Hz, 1H), 6.91 (d, J = 8.9 Hz, 1H), 6.77 (t, J = 5.5 Hz, 1H), 3.38-3.32 (m, 2H), 3.26 (q, J = 6.5 Hz, 2H), 3.22 (s, 3H), 2.65 (s, 6H), 1.65-1.51 (m, 4H).

[0488] 3-(Dimethylsulfamoyl)-4-(6-methoxyhexylamino)benzoic acid (Compound 3.20, Scheme 3).

[0489] The title compound was synthesized according to the general procedure described above using intermediate 3.3 (50 mg, 0.20 mmol) and intermediate 4.4 (53.1, 0.40 mmol) in dry 1,4-dioxane (0.7 ml). Trituration with cyclohexane (1 ml) afforded the pure title compound (23.17 mg, 32% yield). Characterization: Rt = 1.84 min; MS (ESI) m / z: 357.5 [MH]-. Calculated [MH]-: 358.2. 1H NMR (400 MHz, DMSO-d6) δ8.04 (d, J = 2.1 Hz, 1H), 7.93 (dd, J = 8.8, 2.1 Hz, 1H), 6.90 (d, J = 8.9 Hz, 1H), 6.73 (t, J = 5.3 Hz, 1H), 3.31-3.26 (m, 2H), 3.26-3.21 (m, 2H), 3.20 (s, 3H), 1.62-1.53 ​​(m, 2H), 1.52-1.43 (m, 2H), 1.39-1.27 (m, 4H).

[0490] 3-(Cyclopentylsulfamoyl)-4-(8,8,8-trifluorooctylamino)benzoic acid (Compound 3.21, Scheme 3).

[0491] The title compound was synthesized in dry 1,4-dioxane (0.6 ml) using Intermediate 3.4 (50 mg, 0.17 mmol) and Intermediate 4.5 (35.1 mg, 0.19 mmol) according to the general procedure described above. Trituration with diethyl ether (1 ml) afforded the pure title compound (31.7 mg, 41% yield). Characterization: Rt = 2.33 min; MS (ESI) m / z: 449.5 [MH]-. Calculated [MH]-: 450.2. NMR (400MHz, chloroform-d) δ8.49 (d, J=2.1Hz, 1H), 8.08 (dd, J=8.8, 2.1Hz, 1H), 6.7 5(d,J=8.9Hz,1H),6.53(s,1H),4.63-4.51(m,1H),3.63-3.53(m,1H),3.25 (t,J=7.1Hz,2H),2.14-2.00(m,2H),1.85-1.75(m,2H),1.74-1.65(m,2H), 1.65-1.54(m,4H),1.53-1.47(m,2H),1.46-1.36(m,6H),1.36-1.27(m,2H).

[0492] 3-(Cyclohexylsulfamoyl)-4-(8,8,8-trifluorooctylamino)benzoic acid (Compound 3.22, Scheme 3).

[0493] The title compound was synthesized in dry 1,4-dioxane (0.55 ml) using Intermediate 3.5 (50 mg, 0.16 mmol) and Intermediate 4.5 (33.4 mg, 0.18 mmol) according to the general procedure described above. Trituration with diethyl ether (1 ml) afforded the pure title compound (25.3 mg, 34% yield). Characterization: Rt = 2.40 min; MS (ESI) m / z: 463.5 [MH]-. Calculated [MH]-: 464.2. 1H NMR (400 MHz, CHLOROFORM-d) δ 8.49 (d, J = 2.1 Hz, 1H), 8.07 (dd, J = 8.8, 2.1 Hz, 1H), 6.74 (d, J = 8.9 Hz, 1H), 6.50 (s, 1H), 4.49 (d, J = 7.9 Hz, 1H), 3.25 (t, J = 7.1 Hz, 2H), 3.18-3.07 (m, 1H), 2.14-2.00 (m, 2H), 1.79-1.66 (m, 4H), 1.66-1.49 (m, 6H), 1.48-1.34 (m, 6H), 1.30-1.19 (m, 3H), 1.18-1.07 (m, 2H).

[0494] General Procedure I for the Synthesis of Intermediates 4.2-4.3 (Reaction I, Scheme 3) .

[0495] At room temperature, a suspension of potassium phthalimide 4.1 (1 mmol) and an appropriate alkyl bromide (1.2 mmol) in dry N,N-dimethylformamide (3.5 ml) was stirred for 15 hours. After the reaction was complete, the mixture was diluted with water (35 ml) and EtOAc (35 ml). The organic layer was dried over Na2SO4 and concentrated to dryness under low pressure. Finally, the pure title compound was purified by flash chromatography on silica gel.

[0496] 2-(6-Methoxyhexyl)isoindoline-1,3-dione (Compound 4.2, Scheme 4).

[0497] The title compound was synthesized according to the previously described General Procedure I using potassium phthalimide 4.1 (300 mg, 1.60 mmol) and 1-bromo-6-methoxyhexane (0.36 ml, 2.08 mmol) in dry N,N-dimethylformamide (5.5 ml). Purification by flash chromatography on silica gel (cyclohexane / EtOAc 70:30) afforded the pure title compound (355.72 mg, 84% yield). Characterization: Rt = 2.23 min; MS (ESI) m / z: 262.5 [MH]+. Calculated [MH]-: 261.1. 1H NMR (400 MHz, CHLOROFORM-d) δ 7.86-7.79 (m, 2H), 7.73-7.66 (m, 2H), 3.67 (t, J = 7.4 Hz, 2H), 3.34 (t, J = 6.5 Hz, 2H), 3.30 (s, 3H), 1.68 (p, J = 6.1, 5.6 Hz, 2H), 1.56 (p, J = 6.6 Hz, 2H), 1.43-1.31 (m, 4H).

[0498] 2-(8,8,8-Trifluorooctyl)isoindoline-1,3-dione (Compound 4.3, Scheme 4).

[0499] The title compound was synthesized according to the previously described General Procedure I using potassium phthalimide 4.1 (300 mg, 1.60 mmol) and intermediate 8-bromo-1,1,1-trifluorooctane (0.4 ml, 2.08 mmol) in dry N,N-dimethylformamide (5.5 ml). Purification by flash chromatography on silica gel (cyclohexane / EtOAc 85:15) afforded the pure title compound (392.63 mg, 75% yield). Characterization: Rt = 1.76 min; MS (ESI) m / z: 314.4 [MH]+. Calculated [MH]-: 313.1. 1H NMR (400 MHz, CHLOROFORM-d) δ 7.86-7.81 (m, 2H), 7.73-7.67 (m, 2H), 3.70-3.65 (m, 2H), 2.11-1.97 (m, 2H), 1.68 (p, J = 7.2 Hz, 2H), 1.58-1.47 (m, 2H), 1.39-1.30 (m, 6H).

[0500] General Procedure J for the synthesis of compounds 4.4-4.5 (Reaction J, Scheme 4).

[0501] The corresponding intermediate 4.2 or 4.3 (1 mmol) was refluxed in anhydrous ethanol (1.2 mmol) together with hydrazine hydrate (1.5 mmol) for 4 hours. After the reaction was complete, the mixture was cooled to room temperature and the resulting precipitated solid was filtered. The solid was washed with ethanol and the filtrate was concentrated to dryness under low pressure. Finally, the pure title amine was purified by flash chromatography on basic alumina.

[0502] 6-Methoxyhexan-1-amine (Compound 4.4, Scheme 4).

[0503] The title compound was synthesized according to the previously described general procedure J using intermediate 4.2 (356 mg, 1.35 mmol) and hydrazine hydrate (0.15 ml, 2.02 mmol) in anhydrous ethanol (5.5 ml). Purification by flash chromatography on basic alumina (dichloromethane / methanol 90:10) afforded the pure title compound (127.55 mg, 7% yield). Characterization: Rt = 1.00 min; MS (ESI) m / z: 132.4 [MH]+. Calculated [MH]-: 131.1. 1H NMR (400 MHz, DMSO-d6) δ 3.29 (t, J = 6.5 Hz, 2H), 3.20 (s, 3H), 1.51-1.43 (m, 2H), 2.68 (p, J = 6.2 Hz, 2H), 1.37-1.21 (m, 6H).

[0504] 8,8,8-Trifluorooctan-1-amine (Compound 4.5, Scheme 4).

[0505] The title compound was synthesized according to the previously described General Procedure J using intermediate 4.3 (393 mg, 1.24 mmol) and hydrazine hydrate (0.14 ml, 1.86 mmol) in anhydrous ethanol (5.5 ml). Purification by flash chromatography on basic alumina (dichloromethane / methanol 95:5) afforded the pure title compound (136.31 mg, 60% yield). Characterization: Rt = 1.59 min; MS (ESI) m / z: 184.4 [MH]+. Calculated [MH]-: 183.1. 1H NMR (400 MHz, DMSO-d6) δ 2.78-2.68 (m, 2H), 2.30-2.15 (m, 2H), 1.61-1.41 (m, 4H), 1.38-1.21 (m, 6H).

[0506] General Procedure K for the synthesis of compounds 5.2-5.4 (Scheme 5).

[0507] The 4-fluoro-3-chlorosulfonyl-benzoic acid 3.1 (1mmol) that is dissolved in the 2mL THF is dropwise added in the ice-cold solution of 8mL suitable cyclamine (3mmol) in THF, and stirred 1 hour under RT.After reaction is completed, reaction mixture is evaporated to dry, and resistates is handled with water and HCl.Precipitated product is filtered and rinsed with water to obtain pure title compound.

[0508] 4-Fluoro-3-pyrrolidin-1-ylsulfonyl-benzoic acid (Compound 5.2, Scheme 5).

[0509] The title compound was synthesized according to the general procedure K described above using intermediate 3.1 (250 mg, 1.04 mmol) and pyrrolidine (0.26 ml, 3.11 mmol) in THF (8 ml). The described workup afforded the pure title compound (243.2 mg, 85% yield). Characterization: Rt = 1.17 min; MS (ESI) m / z: 272.4 [MH]-. Calculated [MH]-: 273.05. 1H NMR (400 MHz, DMSO-d6) δ 8.30 (dd, J = 6.8, 2.3 Hz, 1H), 8.25 (ddd, J = 8.6, 4.8, 2.3 Hz, 1H), 7.62 (dd, J = 10.1, 8.6 Hz, 1H), 3.28-3.21 (m, 4H), 1.81-1.73 (m, 4H).

[0510] 4-Fluoro-3-(1-piperidinylsulfonyl)benzoic acid (Compound 5.3, Scheme 5).

[0511] The title compound was synthesized according to the general procedure K described above using intermediate 3.1 (250 mg, 1.04 mmol) and piperidine (0.31 ml, 3.11 mmol) in THF (8 ml). The described workup afforded the pure title compound (257.3 mg, 86% yield). Characterization: Rt = 1.34 min; MS (ESI) m / z: 286.4 [MH]-. Calculated [MH]-: 287.06. 1H NMR (400 MHz, DMSO-d6) δ 8.28-8.23 (m, 2H), 7.65-7.58 (m, 1H), 3.08 (t, J = 5.4 Hz, 4H), 1.58-1.49 (m, 4H), 1.46-1.39 (m, 2H).

[0512] 4-Fluoro-3-morpholinosulfonyl-benzoic acid (Compound 5.4, Scheme 5).

[0513] The title compound was synthesized according to the general procedure K described above using intermediate 3.1 (250 mg, 1.04 mmol) and morpholine (0.27 ml, 3.11 mmol) in THF (8 ml). The described workup afforded the pure title compound (248.1 mg, 83% yield). Characterization: Rt = 1.03 min; MS (ESI) m / z: 288.4 [MH]-. Calculated [MH]-: 289.04. 1H NMR (400 MHz, DMSO-d6) δ 8.32-8.24 (m, 2H), 7.64 (dd, J = 10.1, 8.5 Hz, 1H), 3.67-3.60 (m, 4H), 3.10-3.04 (m, 4H).

[0514] 3-Pyrrolidin-1-ylsulfonyl-4-(8,8,8-trifluorooctylamino)benzoic acid (Compound 5.5, Scheme 5).

[0515] The title compound was synthesized in dry 1,4-dioxane (0.55 ml) using Intermediate 5.2 (50 mg, 0.17 mmol) and Intermediate 4.5 (34.8 mg, 0.19 mmol) according to the general procedure described above. Trituration with diethyl ether (1 ml) afforded the pure title compound (17.3 mg, 23% yield). Characterization: Rt = 2.30 min; MS (ESI) m / z: 435.5 [MH]-. Calculated [MH]-: 436.2. 1H NMR (400 MHz, DMSO-d6) δ8.11 (d, J = 2.1 Hz, 1H), 7.92 (dd, J = 8.8, 2.1 Hz, 1H), 6.89 (d, J = 8.9 Hz, 1H), 6.74 (t, J = 5.3 Hz, 1H), 3.24 (q, J = 6.7 Hz, 2H), 3.18-3.11 (m, 4H), 2.29-2.14 (m, 2H), 1.79-1.68 (m, 4H), 1.57 (m, 2H), 1.46 (m =, 2H), 1.33 (s, 6H).

[0516] 3-(1-Piperidinylsulfonyl)-4-(8,8,8-trifluorooctylamino)benzoic acid (Compound 5.6, Scheme 5).

[0517] The title compound was synthesized in dry 1,4-dioxane (0.55 ml) using Intermediate 5.3 (50 mg, 0.17 mmol) and Intermediate 4.5 (34.8 mg, 0.19 mmol) according to the general procedure described above. Trituration with diethyl ether (1 ml) afforded the pure title compound (13 mg, 17% yield). Characterization: Rt = 2.40 min; MS (ESI) m / z: 449.5 [MH]-. Calculated [MH]-: 450.2. 1H NMR (400 MHz, DMSO-d6) δ8.04 (d, J = 2.1 Hz, 1H), 7.92 (dd, J = 8.8, 2.1 Hz, 1H), 6.89 (d, J = 9.0 Hz, 1H), 6.69 (t, J = 5.4 Hz, 1H), 3.24 (q, J = 6.7 Hz, 2H), 2.98 (t, J = 5.4 Hz, 4H), 2.29-2.15 (m, 2H), 1.62-1.55 (m, 2H), 1.55-1.43 (m, 6H), 1.42-1.37 (m, 2H), 1.37-1.30 (m, 6H).

[0518] 3-Morpholinosulfonyl-4-(8,8,8-trifluorooctylamino)benzoic acid (Compound 5.7, Scheme 5).

[0519] The title compound was synthesized in dry 1,4-dioxane (0.55 ml) using Intermediate 5.4 (50 mg, 0.17 mmol) and Intermediate 4.5 (34.8 mg, 0.19 mmol) according to the general procedure described above. Trituration with diethyl ether (1 ml) afforded the pure title compound (28.4 mg, 37% yield). Characterization: Rt = 2.21 min; MS (ESI) m / z: 451.2 [MH]-. Calculated [MH]-: 452.16. 1H NMR (400 MHz, CHLOROFORM-d) δ 8.33 (d, J = 2.1 Hz, 1H), 8.07 (dd, J = 8.9, 2.1 Hz, 1H), 6.87 (t, J = 5.0 Hz, 1H), 6.74 (d, J = 9.0 Hz, 1H), 3.77-3.70 (m, 4H), 3.21 (q, J = 7.0 Hz, 2H), 3.12-3.06 (m, 4H), 2.14-1.99 (m, 2H), 1.73-1.63 (m, 2H), 1.61-1.50 (m, 2H), 1.48-1.32 (m, 6H).

[0520] 5-Cyano-2-fluoro-N,N-dimethyl-benzenesulfonamide (Compound 6.2, Reaction L, Scheme 6).

[0521] 5-cyano-2-fluorobenzene-1-sulfonyl chloride 6.1 (300mg, 1.35mmol) dissolved in 3.5mL THF is added dropwise to 2M dimethylamine in THF (0.74ml, 1.49mmol) and N, N-diisopropylethylamine (0.48ml, 2.70mmol) in an ice-cold solution in 10ml THF, then stirred at room temperature for 30 minutes. After the reaction is complete, the reaction mixture is evaporated to dryness, and the residue is distributed between ethyl acetate (50ml) and water (50ml), and each layer is separated. Use Na2SO4 dry organic layer and be concentrated to dryness under low pressure. By flash chromatography on silica gel (cyclohexane / DCM+1% EtOAc70:30 to 30:70), pure title compound (194.2mg, yield 63%) is obtained. Characterization: 1H NMR (400 MHz, chloroform-d) δ 8.20 (dd, J = 6.2, 2.2 Hz, 1H), 7.87 (ddd, J = 8.6, 4.4, 2.2 Hz, 1H), 7.36 (t, J = 8.9 Hz, 1H), 2.89 (d, J = 1.9 Hz, 6H).

[0522] 5-Cyano-N,N-dimethyl-2-(8,8,8-trifluorooctylamino)benzenesulfonamide (Compound 6.3, Scheme 6).

[0523] The title compound was synthesized according to the general procedure described above using intermediate 6.2 (194 mg, 0.84 mmol) and intermediate 4.5 (311.5 mg, 1.64 mmol) in dry 1,4-dioxane (4.2 ml). Trituration with diethyl ether (3 ml) provided the pure title compound (317.2 mg, 97% yield). Characterization: Rt = 1.82 min; MS (ESI) m / z: 390.3 [MH]-. Calculated [MH]-: 391.15. 1H NMR (400 MHz, CHLOROFORM-d) δ 7.87 (d, J = 2.0 Hz, 1H), 7.57 (dd, J = 8.8, 2.1 Hz, 1H), 6.85 (s, 1H), 6.72 (d, J = 8.8 Hz, 1H), 3.23-3.13 (m, 2H), 2.77 (s, 6H), 2.14-1.98 (m, 2H), 1.73-1.61 (m, 2H), 1.60-1.48 (m, 4H), 1.46-1.33 (m, 6H).

[0524] 4-Fluoro-2-hydroxy-5-sulfamoyl-benzoic acid (Compound 7.3, Reaction M, Scheme 7)

[0525] 4-Fluoro-2-hydroxy-benzoic acid 7.1 (2 g, 12.81 mmol) was stirred in chlorosulfonic acid (4.30 ml, 64.06 mmol) at 120°C for 4 hours. After the reaction was complete, the mixture was slowly poured into ice-cold water (50 ml), and the resulting precipitated solid was collected by filtration to provide intermediate 7.2. This intermediate (1.12 g, 4.35 mmol) was quickly dissolved in 10 ml of THF and added to an ice-cold solution of 0.83 ml of 20% aqueous NH₄OH (4.35 mmol) and trimethylamine (0.61 ml, 4.34 mmol) in 30 ml of tetrahydrofuran. The reaction mixture was stirred at 0°C for 8 hours. After the reaction was complete, the mixture was evaporated to dryness under reduced pressure, and the residue was treated with saturated aqueous NH₄Cl (50 ml) and extracted twice with EtOAc (2 x 50 ml). The combined organic phases were dried over Na₂SO₄ and concentrated to dryness under reduced pressure to yield the pure title compound (915.9 mg, 30% yield over two steps). Characterization: Rt = 1.15 min; MS (ESI) m / z: 234.3 [MH]-. Calculated [MH]-: 235. 1H NMR (400 MHz, DMSO-d₆) δ 8.21 (d, J = 8.5 Hz, 1H), 7.61 (s, 2H), 7.03 (d, J = 11.7 Hz, 1H).

[0526] 2-Hydroxy-5-sulfamoyl-4-(8,8,8-trifluorooctylamino)benzoic acid (Compound 7.4, Scheme 7).

[0527] The title compound was synthesized according to the general procedure described above using intermediate 7.3 (250 mg, 1.02 mmol) and intermediate 4.5 (377.7 mg, 2.04 mmol) in dry 1,4-dioxane (3.4 ml). Trituration with cyclohexane (3 ml) afforded the pure title compound (286 mg, 69% yield). Characterization: Rt = 1.81 min; MS (ESI) m / z: 397.3 [MH]-. Calculated [MH]-: 398.1. 1H NMR (400 MHz, DMSO-d6) δ 8.10 (s, 1H), 7.32 (s, 2H), 6.36 (t, J = 5.3 Hz, 1H), 6.12 (s, 1H), 3.18 (q, J = 6.8 Hz, 2H), 2.29-2.15 (m, 2H), 1.64-1.54 (m, 2H), 1.52-1.42 (m, 2H), 1.41-1.29 (m, 6H).

[0528] Tert-butyl 4-(5,5,5-trifluoropentyl)piperazine-1-carboxylate (Compound 8.2, Reaction N, Scheme 8).

[0529] To a solution of 1-boc-piperazine 8.1 (400 mg, 2.15 mmol) in acetonitrile (5 mL) cooled at 0 ° C, 5-iodo-1,1,1-trifluoropentane (0.25 mL, 3.22 mmol) and N,N-diisopropylethylamine (0.57 mL, 3.22 mmol) were added, and the reaction mixture was stirred at room temperature for 24 hours. After the reaction was completed, the crude reaction product was concentrated to dryness under low pressure. The residue was dissolved in EtOAc (25 mL) and washed with water (25 mL) and brine (25 mL). The organic layer was dried over Na2SO4 and concentrated to dryness under low pressure. Purification by silica gel flash chromatography (dichloromethane / methanol 98:2) gave the pure title compound (378.9 mg, 92% yield). Characterization: Rt = 2.02; MS (ESI) m / z: 311.5 [MH]+. Calculated [MH]-: 310.2. 1H NMR (400 MHz, CHLOROFORM-d) δ 3.42 (t, J = 4.7 Hz, 4H), 2.41-2.31 (m, 6H), 2.16-2.02 (m, 2H), 1.63-1.50 (m, 4H), 1.45 (s, 9H).

[0530] 1-(5,5,5-Trifluoropentyl)piperazine di-trifluoroacetate (Compound 8.3, Reaction O, Scheme 8)

[0531] Intermediate 8.2 (378.9 mg, 2.01 mmol) was stirred in pure trifluoroacetic acid (1.5 mL) at room temperature for 1.5 hours. After completion of the reaction, the crude product was diluted with DCM and concentrated to dryness three times (3x10 ml) at low pressure, and diluted with MeOH (10 ml) and concentrated to dryness once at low pressure to give the pure title compound (717.5 mg, 81% yield). Characterization: 1H NMR (400 MHz, methanol-d4) δ 3.59-3.48 (m, 8H), 3.31-3.28 (m, 2H), 3.22-3.15 (m, 2H), 2.30-2.17 (m, 2H), 1.87-1.78 (m, 2H), 1.68-1.59 (m, 2H).

[0532] 3-(Dimethylsulfamoyl)-4-[4-(5,5,5-trifluoropentyl)piperazin-1-yl]benzoic acid (Compound 9.1, Reaction P, Scheme 9)

[0533] Under an argon atmosphere, intermediate 8.3 (106.4 mg, 0.24 mmol) and triethylamine (0.14 ml, 1.00 mmol) in dry 1,4-dioxane (1 ml) were added intermediate 3.3 (50 mg, 0.20 mmol) dissolved in 1,4-dioxane (1 ml), and the reaction mixture was stirred at 100°C for 24 hours. After completion of the reaction, the crude reaction product was partitioned between ethyl acetate (25 ml) and saturated NH4Cl solution (25 ml), and the pH was adjusted to 3 with concentrated HCl. The layers were separated and the aqueous layer was washed with diethyl ether (25 ml). The aqueous layer was then neutralized to pH 7 and extracted with ethyl acetate (3 x 25 ml) and DCM (25 ml). The combined organic layers were dried over Na2SO4 and concentrated to dryness under low pressure. Trituration with diethyl ether (2 ml) gave the pure title compound (26.7 mg, 30% yield). Characterization: Rt = 1.31; MS (ESI) m / z: 436.5 [MH]-. Calculated [MH]-: 437.2. 1H NMR (400 MHz, DMSO-d6) δ 8.33 (d, J = 2.1 Hz, 1H), 8.12 (dd, J = 8.3, 2.2 Hz, 1H), 7.56 (d, J = 8.4 Hz, 1H), 3.08-2.99 (m, 4H), 2.67 (s, 6H), 2.57-2.53 (m, 4H), 2.40-2.34 (m, 2H), 2.34-2.18 (m, 2H), 1.58-1.46 (m, 4H).

[0534] N,N-dimethyl-5-(1H-tetrazol-5-yl)-2-(8,8,8-trifluorooctylamino)benzenesulfonamide (Compound 10.1, Scheme 10, Figure 12 ).

[0535] A mixture of intermediate 6.3 (317.2 mg, 0.8 mmol), sodium azide (63.2 mg, 0.96 mmol) and zinc chloride (132.6 mg, 0.96 mmol) was stirred in 4 ml of n-butanol at 110 ° C for 10 hours. After the reaction was completed, the reaction mixture was evaporated to dryness under low pressure. Next, 5% NaOH (20 mL) was added and the mixture was stirred for 20 min. The resulting suspension was filtered and the solid was washed with 5% NaOH (10 mL). The pH of the filtrate was adjusted to 1.0 with concentrated HCl and extracted 3 times with EtOAc (3x25 ml). The combined organic layers were dried over Na2SO4 and concentrated to dryness under low pressure. Finally, purification by silica gel flash chromatography (dichloromethane / methanol 98:2) provided the pure title compound (110.93 mg, yield 32%). Characterization: Rt = 0.77; MS (ESI) m / z: 433.3 [MH]-. Calculated [MH]-: 434.2. 1H NMR (400 MHz, CHLOROFORM-d) δ 8.25 (d, J = 2.1 Hz, 1H), 8.19 (dd, J = 8.8, 2.2 Hz, 1H), 6.85 (d, J = 8.9 Hz, 1H), 6.61 (s, 1H), 3.19 (t, J = 7.1 Hz, 2H), 2.76 (s, 6H), 2.14-1.98 (m, 2H), 1.73-1.62 (m, 2H), 1.61-1.49 (m, 2H), 1.49-1.30 (m, 6H).

[0536] 5-(N,N-dimethylsulfamoyl)-4-fluoro-2-hydroxybenzoic acid (Compound 12.1, Scheme 12). 4-Fluoro-2-hydroxy-benzoic acid 7.1 (2 g, 12.81 mmol) was stirred in chlorosulfonic acid (4.30 ml, 64.06 mmol) at 120 ° C for 4 hours. After the reaction was completed, the mixture was slowly poured into ice-cold water (50 ml) and the resulting precipitated solid was collected by filtration. The collected solid (1.141 g) was dissolved in 10 ml THF and added dropwise to an ice-cold solution of 2M dimethylamine in THF (3 ml) and DIPEA (3 ml) in 35 ml tetrahydrofuran. The reaction mixture was stirred at 0 ° C for 8 hours. After the reaction was completed, the mixture was evaporated to dryness under low pressure, and the residue was treated with saturated NH4Cl solution (50 ml) and extracted twice with EtOAc (2x50 ml). The combined organic layers were dried over Na2SO4 and concentrated to dryness under low pressure to afford the pure title compound (823.9 mg, 70% yield). UPLC / MS: Rt = 1.19 min (step 1); MS (ESI) m / z: 262.0 [MH] - Calculated [MH] - :262.0. 1H NMR (400 MHz, DMSO-d 6) δ8.15(d,J=8.2Hz,1H),7.13-7.03(m,1H),2.71(d,J=1.7Hz,6H).

[0537] 5-(N, N-dimethylsulfamoyl)-4-fluoro-2-methoxybenzoic acid methyl ester (compound 12.2, scheme 12).To the ice-cold solution of intermediate 12.1 (200mg, 0.75mmol) in DCM / MeOH 8:2 (9ml), trimethylsilyldiazomethane (2M in hexane, 1.13ml, 2.26mmol) was carefully added, and the reaction mixture was stirred for 2 hours at room temperature. After the completion of the reaction, the reaction mixture was quenched with the 1M acetic acid solution in 2ml methanol and evaporated to dryness.The dry residue was suspended in saturated NaHCO (15ml) aqueous solution and extracted twice (2x15ml) with EtOAc. By flash chromatography on silica gel (cyclohexane / EtOAc, from 85:15 to 70:30), there is provided pure title compound (201mg, 92% yield), as a white solid. UPLC / MS: Rt=1.75 min (step 1); MS (ESI) m / z: 292.1 [M+H] + Calculated [M+H] + :292.0. 1 HNMR (600 MHz, chloroform-d) δ 8.35 (d, J = 5.0 Hz, 1H), 6.94 (d, J = 8.0 Hz, 1H), 3.85 (s, 3H), 3.79 (s, 3H), 2.72 (s, 6H).

[0538] Methyl 5-(N,N-dimethylsulfamoyl)-2-methoxy-4-((8,8,8-trifluorooctyl)amino)benzoate (Compound 12.3, Scheme 12). Compound 12.3 was synthesized according to the general procedure described previously using intermediate 12.2 (50 mg, 0.17 mmol) and intermediate 4.5 (75.4 mg, 0.34 mmol) in dry 1,4-dioxane (0.85 ml). Purification by silica gel flash chromatography (cyclohexane / EtOAc, from 80:15 to 75:25) gave the pure title compound (64.9 mg, 84% yield) as a white solid. UPLC / MS: Rt = 2.65 min (step 1); MS (ESI) m / z: 455.3 [M+H] + Calculated [M+H] + :455.2. 1H NMR (400MHz, chloroform-d) δ8.23(s,1H),6.77(t,J=4.8Hz,1H),6.10(s,1H),3.97(s,3H),3.84(s,3H),3.22-3.16(m,2H),2.75(s, 6H), 2.14-2.04 (m, 2H), 1.72 (p, J=7.1Hz, 2H), 1.60-1.55 (m, 4H), 1.45 (dd, J=5.0, 2.0Hz, 2H), 1.41 (dd, J=3.9, 2.6Hz, 4H).

[0539] Methyl 5-(N,N-dimethylsulfamoyl)-2-hydroxy-4-((8,8,8-trifluorooctyl)amino)benzoate (Compound 12.4, Scheme 12). Under argon atmosphere, to intermediate 12.3 (50 mg, 0.11 mmol) dissolved in DCM (1.2 mL) was added dropwise BBr (1 M in DCM, 0.55 ml, 0.55 mmol) and the mixture was stirred at room temperature for 6 hours. After completion of the reaction, the reaction mixture was cooled to 0 ° C, quenched with 2 ml of methanol and evaporated to dryness. The dried crude residue was then partitioned between EtOAc (10 ml) and saturated NH4Cl solution (10 ml), and the layers were separated. The organic layer was dried over Na2SO4 and concentrated to dryness under low pressure. Purification by flash chromatography on silica gel (cyclohexane / EtOAc 95:05) gave the pure title compound (40.2 mg, 83% yield) as a white solid. UPLC / MS: Rt = 2.10 min (step 1); MS (ESI) m / z: 441.3 [MH] + Calculated [M+H] + :441.1. 1 H NMR (400 MHz, CHLOROFORM-d) δ 11.26 (s, 1H), 8.17 (s, 1H), 6.73 (t, J = 4.6 Hz, 1H), 6.16 (s, 1H), 3.92 (s, 3H), 3.16 (q, J = 7.1, 5.0 Hz, 2H), 2.75 (s, 6H), 2.15-1.99 (m, 2H), 1.74-1.63 (m, 2H), 1.62-1.54 (m, 2H), 1.48-1.35 (m, 6H).

[0540] Methyl 5-(N,N-dimethylsulfamoyl)-2-ethoxy-4-((8,8,8-trifluorooctyl)amino)benzoate (Compound 12.5, Scheme 12). To a solution of intermediate 12.4 (31.8 mg, 0.07 mmol) in acetonitrile (0.7 mL) was added iodoethane (10 μl, 0.11 mmol) and potassium carbonate (15 mg, 0.11 mmol), and the reaction mixture was stirred at 80 ° C for 10 hours. After completion of the reaction, the crude product was partitioned between EtOAc (10 ml) and water (10 ml), and the layers were separated. The organic layer was dried over Na2SO4 and concentrated to dryness under low pressure. Purification by flash chromatography on silica gel (cyclohexane / EtOAc, from 100:00 to 80:20) gave the pure title compound (25.6 mg, 78% yield) as a white solid. UPLC / MS: Rt=1.85 min (step 1); MS (ESI) m / z: 469.3 [M+H] + Calculated [M+H] + :469.2. 1 H NMR (400MHz, chloroform-d) δ8.20(s,1H),6.71(t,J=4.8Hz,1H),6.07(s,1H),4.14(q,J=7.0Hz,2H),3.82(s,3H),3.18-3.11(m ,2H),2.72(s,6H),2.13-1.99(m,2H),1.73-1.64(m,2H),1.61-1.53(m,2H),1.51(t,J=6.9Hz,3H),1.48-1.35(m,6H).

[0541] 5-(N,N-Dimethylsulfamoyl)-2-ethoxy-4-((8,8,8-trifluorooctyl)amino)benzoic acid (Compound 12.7, Scheme 12). To a solution of compound 12.5 (25.6 mg, 0.05 mmol) in tetrahydrofuran (0.5 mL) was added 1 M aqueous LiOH (0.27 mL, 0.27 mmol), and the reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction, the crude product was partitioned between EtOAc (10 mL) and saturated NH4Cl solution (10 mL), and the layers were separated. The organic layer was dried over Na2SO4 and concentrated to dryness under reduced pressure. Trituration with cyclohexane afforded the pure title compound (19.54 mg, 86% yield) as a white solid. UPLC / MS: Rt = 1.32 min (step 1); MS (ESI) m / z: 453.3 [MH] - Calculated [MH] - :453.2. 1 H NMR (400 MHz, DMSO-d6) δ7.95(s,1H),6.62(t,J=5.2Hz,1H),6.23(s,1H),4.15(q,J=6.9Hz,2H),3.23(q,J=6.5Hz,2 H),2.60(s,6H),2.29-2.14(m,2H),1.63-1.52(m,2H),1.51-1.42(m,2H),1.40-1.25(m,9H).

[0542] Methyl 2-(cyclopentyloxy)-5-(N,N-dimethylsulfamoyl)-4-((8,8,8-trifluorooctyl)amino)benzoate (Compound 12.6, Scheme 12). To a solution of intermediate 12.4 (30.0 mg, 0.07 mmol) in acetonitrile (0.7 mL) was added cyclopentyl bromide (15 μl, 0.13 mmol) and potassium carbonate (28.3 mg, 0.20 mmol), and the reaction mixture was stirred at 80 °C for 4 hours. After completion of the reaction, the crude product was partitioned between EtOAc (10 ml) and water (10 ml), and the layers were separated. The organic layer was dried over Na2SO4 and concentrated to dryness under low pressure. Purification by flash chromatography on silica gel (cyclohexane / EtOAc, from 100:00 to 90:10) gave the pure title compound (25.6 mg, 72% yield) as a white solid. UPLC / MS: Rt=2.30 min (step 2); MS (ESI) m / z: 509.2 [M+H] + Calculated [M+H] + :509.6. 1 H NMR (400 MHz, chloroform-d) δ 8.19 (s, 1H), 6.69 (t, J = 4.8 Hz, 1H), 6.07 (s, 1H), 4.88-4.81 (m, 1H), 3.80 (s, 3H), 3.19-3.10 (m, 2H), 2.72 (s, 6H), 2.13-1.99 (m, 2H), 1.99-1.92 (m, 4H), 1.91-1.81 (m, 2H), 1.73-1.62 (m, 2H), 1.61-1.51 (m, 2H), 1.49-1.34 (m, 6H).

[0543] 2-(Cyclopentyloxy)-5-(N,N-dimethylsulfamoyl)-4-((8,8,8-trifluorooctyl)amino)benzoic acid (Compound 12.8, Scheme 12). To a solution of intermediate 12.6 (25.6 mg, 0.05 mmol) in tetrahydrofuran (0.25 mL) was added 1 M aqueous LiOH (0.5 mL, 0.25 mmol), and the mixture was stirred at room temperature for 16 hours. After completion of the reaction, the crude product was partitioned between EtOAc (10 mL) and saturated NH4Cl solution (10 mL), and the layers were separated. The organic layer was dried over Na2SO4 and concentrated to dryness under reduced pressure. Trituration with cyclohexane afforded the pure title compound (16.3 mg, 66% yield) as a white solid. UPLC / MS: Rt = 1.80 min (step 1); MS (ESI) m / z: 493.3 [MH] - Calculated [MH] - :493.2. 1 HNMR (400 MHz, chloroform-d) 1 H NMR (400 MHz, CHLOROFORM-d) δ 8.40 (s, 1H), 6.94 (s, 1H), 6.12 (s, 1H), 5.09-5.03 (m, 1H), 3.20-3.13 (m, 2H), 2.75 (s, 6H), 2.14-1.97 (m, 5H), 1.93-1.81 (m, 2H), 1.81-1.65 (m, 4H), 1.61-1.51 (m, 4H), 1.50-1.33 (m, 6H).

[0544] 5-(N,N-Dimethylsulfamoyl)-2-methoxy-4-((8,8,8-trifluorooctyl)amino)benzoic acid (Compound 13.1, Scheme 13). To a solution of intermediate 12.3 (59 mg, 0.13 mmol) in tetrahydrofuran (1.3 mL) was added 1 M aqueous LiOH (0.26 mL, 0.26 mmol), and the mixture was stirred at room temperature for 16 hours. After completion of the reaction, the crude product was partitioned between EtOAc (10 mL) and saturated NH4Cl solution (10 mL), and the layers were separated. The organic layer was dried over Na2SO4 and concentrated to dryness under reduced pressure. Trituration with cyclohexane gave the pure title compound (41.2 mg, 72% yield) as a white solid. UPLC / MS: Rt = 1.16 min (step 1); MS (ESI) m / z: 439.5 [MH] - Calculated [MH] - :439.2. 1H NMR(400MHz,DMSO-d6)δ7.98(s,1H),6.65(t,J=5.2Hz,1H),6.26(s,1H),3.88(s,3H),3 .29-3.22(m,2H),2.61(s,6H),1.65-1.55(m,2H),1.52-1.42(m,4H),1.39-1.29(m,6H).

[0545] 4-Fluoro-3-(N-(tetrahydro-2H-pyran-4-yl)sulfamoyl)benzoic acid (Compound 14.1, Scheme 14). The title compound was synthesized according to General Procedure G described previously using Intermediate 3.1 (250 mg, 1.04 mmol) and tetrahydro-2H-pyran-4-amine (0.32 ml, 2.07 mmol) in THF (8.5 ml). The described workup afforded the pure title compound (160.9 mg, 51% yield) as a white solid. UPLC / MS: Rt = 0.93 min (step 1); MS (ESI) m / z: 302.1 [MH] - . Calculated [MH]: 302.06. 1 H NMR (400MHz, DMSO-d6) δ8.34(dd,J=7.1,2.3Hz,1H),8.27(d,J=7.8Hz,1H),8.24-8.18(m,1H),7. 57(t,J=9.3Hz,1H),3.77-3.68(m,2H),3.27-3.19(m,3H),1.58-1.49(m,2H),1.49-1.37(m,2H).

[0546] 3-((4,4-Difluoropiperidin-1-yl)sulfonyl)-4-fluorobenzoic acid (Compound 14.2, Scheme 14). The title compound was synthesized according to the general procedure K described above using intermediate 3.1 (150 mg, 0.62 mmol) and 4,4-difluoropiperidine hydrochloride (198.1 mg, 1.24 mmol) and DIPEA (0.33 ml, 1.87 mmol) in THF (5.0 ml). After completion of the reaction, the reaction mixture was evaporated to dryness. The described workup afforded the pure title compound (176.4 mg, 88% yield) as a white solid. UPLC / MS: Rt = 1.38 min (step 1); MS (ESI) m / z: 322.0 [MH] - . [MH] calculated: 322.04. 1H NMR (400MHz, DMSO-d6) δ8.31-8.25(m,2H),7.67-7.60(m,1H),3.29(t,J=5.8Hz,4H),2.07(ddd,J=19.7,13.7,5.8Hz,4H).

[0547] 3-Morpholinosulfonyl-4-((8,8,8-trifluorooctyl)amino)benzoic acid (Compound 14.3, Scheme 14). The title compound was synthesized according to the general procedure described above using Intermediate 14.2 (50 mg, 0.17 mmol) and Intermediate 4.5 (34.8 mg, 0.19 mmol) in dry 1,4-dioxane (0.55 ml). Purification by silica gel flash chromatography (CHCl / MeOH from 100:0 to 98:02) followed by trituration with diethyl ether (1 ml) afforded the pure title compound (28.4 mg, 37% yield) as a white solid. UPLC / MS: Rt = 2.21 min (step 1); MS (ESI) m / z: 451.2 [MH] - Calculated [MH] - :451.2. 1 H NMR (400MHz, chloroform-d) δ8.33(d,J=2.1Hz,1H),8.07(dd,J=8.9,2.1Hz,1H),6.87(t,J=5.0Hz,1H),6.74(d,J=9.0Hz,1H),3.77-3.7 0(m,4H),3.21(q,J=7.0Hz,2H),3.12-3.06(m,4H),2.14-1.99(m,2H),1.73-1.63(m,2H),1.61-1.50(m,2H),1.48-1.32(m,6H).

[0548] 3-((4,4-Difluoropiperidin-1-yl)sulfonyl)-4-((8,8,8-trifluorooctyl)amino)benzoic acid (Compound 14.4, Scheme 14). The title compound was synthesized according to the general procedure described above using Intermediate 14.1 (50 mg, 0.15 mmol) and Intermediate 4.5 (34.8 mg, 0.19 mmol) in dry 1,4-dioxane (0.55 mL). Purification by silica gel flash chromatography (CHCl / MeOH from 100:0 to 98:02) followed by trituration with petroleum ether (1 mL) afforded the pure title compound (22.6 mg, 31% yield) as a white solid. UPLC / MS: Rt = 2.39 min (step 1); MS (ESI) m / z: 485.2 [MH] - Calculated [MH] -:485.2.1H NMR (400MHz, chloroform-d) δ8.35(d,J=2.0Hz,1H),8.08(dd,J=8.9,2.1Hz,1H),6.78(t,J=5.0Hz,1H),6.74(d,J=9.0Hz,1H),3. 31(t,J=5.8Hz,4H),3.25-3.18(m,2H),2.14-2.00(m,6H),1.69(p,J=7.0Hz,2H),1.62-1.52(m,2H),1.49-1.35(m,6H).

[0549] 3-(Dimethylsulfamoyl)-4-(hept-6-enylamino)benzoic acid (Compound 15.1, Scheme 15). The title compound was synthesized according to the general procedure described above using intermediate 3.3 (420 mg, 1.68 mmol) and hept-6-en-1-amine hydrochloride (335.6 mg, 1.68 mmol) in dry 1,4-dioxane (16.5 mL). Purification by silica gel flash chromatography (CHCl / MeOH from 100:0 to 98:02) followed by trituration with diethyl ether (3 mL) afforded the pure title compound (409.6 mg, 72% yield) as a white solid. UPLC / MS: Rt = 2.13 min (step 1); MS (ESI) m / z: 439.2 [MH] - Calculated [MH] - :339.1. 1 H NMR (400MHz, chloroform-d) δ8.34(d,J=2.0Hz,1H),8.06(dd,J=8.9,2.1Hz,1H),6.91(t,J=5.0Hz,1H),6.72(d,J=9.0Hz,1H),5.80(ddt,J=16 .9,10.2,6.7Hz,1H),5.04-4.91(m,2H),3.24-3.18(m,2H),2.77(s,6H),2.13-2.02(m,2H),1.69(p,J=7.0Hz,2H),1.49-1.39(m,4H).

[0550] 3-(N, N-dimethylsulfamoyl)-4-(hept-6-ene-1-ylamino) methyl benzoate (compound 15.2, scheme 12). To the ice-cold solution of intermediate 15.1 (220mg, 0.64mmol) in DCM / MeOH 8:2 (8ml), trimethylsilyldiazomethane (2M in hexane, 0.48ml, 0.96mmol) was carefully added, and the reaction mixture was stirred for 2 hours at room temperature. After the reaction was completed, the reaction mixture was quenched with 2ml of 1M acetic acid solution in methanol and evaporated to dryness. The dried residue was suspended in saturated NaHCO (15ml) aqueous solution and extracted twice (2x15ml) with EtOAc. Purified by flash chromatography on silica gel (cyclohexane / EtOAc, from 100:00 to 90:10), pure title compound (213.2mg, 94% yield) was obtained as a white solid. UPLC / MS: Rt=1.81 min (step 1); MS (ESI) m / z: 355.2 [M+H] + Calculated [M+H] + :355.2. 1 HNMR (600 MHz, chloroform-d) 1 H NMR (400MHz, chloroform-d) δ8.28(d,J=2.1Hz,1H),8.01(dd,J=8.9,2.1Hz,1H),6.83-6.74(m,1H),6.70(d,J=8.9Hz,1H),5.79(ddt,J=16.9,10. 2,6.7Hz,1H),5.04-4.92(m,2H),3.87(s,3H),3.23-3.15(m,2H),2.75(s,6H),2.12-2.03(m,2H),1.74-1.63(m,2H),1.49-1.38(m,4H).

[0551] Methyl 4-((8-bromo-8,8-difluorooctyl)amino)-3-(N,N-dimethylsulfamoyl)benzoate (Compound 15.3, Scheme 15). To a solution of intermediate 15.2 (213.2 mg, 0.62 mmol) in THF (6.2 ml) in a sealed glass tube were added potassium bicarbonate (62.7 mg, 0.62 mmol), eosin salt (23.8 mg 0.03 mmol), and dibromodifluoromethane (0.12 ml, 1.24 mmol). The reaction mixture was then stirred at room temperature under blue LED irradiation (λ = 460-470 nm) for 16 hours. After completion of the reaction, the reaction mixture was evaporated to dryness. The dried residue was suspended in an aqueous solution of water (25 ml) and extracted twice with EtOAc (2 x 25 ml). Purification by flash chromatography on silica gel (petroleum ether / TBME, from 100:00 to 80:20) afforded the pure title compound (144.5 mg, 48% yield) as a white solid. 15). UPLC / MS: Rt = 2.13 min (step 2); MS (ESI) m / z: 485.0 [M+H] + Calculated [M+H] + :485.08. 1 H NMR (600 MHz, chloroform-d) 1 H NMR (400 MHz, chloroform-d) 1H NMR (400 MHz, chloroform-d) δ 8.27 (d, J = 2.1 Hz, 1H), 8.02 (dd, J = 8.9, 2.1 Hz, 1H), 6.79 (t, J = 5.0 Hz, 1H), 6.70 (d, J = 8.9 Hz, 1H), 3.87 (s, 3H), 3.23-3.16 (m, 2H), 2.76 (s, 6H), 2.40-2.26 (m, 2H), 1.72-1.55 (m, 6H), 1.48-1.35 (m, 6H).

[0552] 4-[(8-Bromo-8,8-difluoro-octyl)amino]-3-(dimethylsulfamoyl)benzoic acid (Compound 15.4, Scheme 15). To a solution of intermediate 15.3 (50 mg, 0.10 mmol) in tetrahydrofuran (1.0 mL) was added 1 M aqueous LiOH (0.42 mL, 0.2 mmol), and the mixture was stirred at room temperature for 16 hours. After completion of the reaction, the crude product was partitioned between EtOAc (10 mL) and saturated NH4Cl solution (10 mL), and the layers were separated. The organic layer was dried over Na2SO4 and concentrated to dryness under reduced pressure. Trituration with cyclohexane gave the pure title compound (40.1 mg, 85% yield) as a white solid. UPLC / MS: Rt = 1.22 min (step 2); MS (ESI) m / z: 469.1 [MH]- Calculated [MH] - :469.1. 1 H NMR (400 MHz, chloroform-d) 1H NMR (400 MHz, chloroform-d) δ 8.29 (d, J = 2.1 Hz, 1H), 8.05 (dd, J 8.9, 2.1 Hz, 1H), 6.83 (t, J = 5.0 Hz, 1H), 6.70 (d, J = 8.9 Hz, 1H), 3.25-3.18 (m, 2H), 2.77 (s, 6H), 2.42-2.28 (m, 2H), 1.76-1.59 (m, 6H), 1.51-1.38 (m, 6H).

[0553] Example 2: Activity Data

[0554] Table 1 below reports the data obtained.

[0555] Table 1

[0556]

[0557]

[0558]

[0559]

[0560]

[0561]

[0562]

[0563]

[0564]

[0565]

[0566]

[0567]

[0568]

[0569] According to one embodiment of the invention, the most active compounds are: compounds 1.7, 1.17, 2.2, 2.6, 2.7, 2.8, 2.9, 3.6, 3.7, 3.8, 3.9, 3.10, 3.11, 3.12, 3.13, 3.14, 3.17, 3.20, 3.21, 3.22, 5.5, 5.6, 5.7, 13.1, 14.4, 15.1.

[0570] Chloride kinetics determination

[0571] To screen the efficiency of compounds in blocking NKCC1 in vitro, Cl - Sensitive membrane marker yellow fluorescent protein (mbYFPQS, Addgene) was used to measure Cl in cells. - Functional NKCC1 transporter assay was performed by measuring changes in ion concentration. mbYFPQS fluorescence correlates with intracellular Cl - The concentration of Cl is inversely proportional to - In particular, NKCC1 or a mock construct (control) was used to - HEK293 cells were transfected with YFP. - Cells were treated with bumetanide and furosemide (as positive controls) or with each test compound of the present invention in culture medium. After 30 minutes, the inhibitory activity of the compound was tested by monitoring the fluorescence upon application of NaCl (1a). NaCl is transported by NKCC1 and binds to YFP, resulting in a decrease in fluorescence. Compared to mock-transfected cells, NKCC1 - - Transfected cells showed a significant decrease in fluorescence levels after NaCl application (1b). Preincubation with bumetanide at 10 μM and 100 μM significantly reduced this effect, while preincubation with furosemide was only effective at 100 μM (1b). Furthermore, the data were normalized again due to the decrease in fluorescence observed in mock-transfected cells after application of bumetanide or furosemide. Selected compounds were tested for their NKCC1 inhibitory activity using a Cl kinetics assay (1c). Notably, at 100 μM, compound 3.17 exhibited superior inhibition of NKCC1 compared to both bumetanide and furosemide.

[0572] Calcium kinetics assay

[0573] Next, the ability of the compounds of the invention to restore depolarizing GABAergic signaling in immature neurons was tested. This effect was indirectly measured as calcium influx into cells by an in vitro calcium kinetics assay in primary cultures of hippocampal neurons. The calcium kinetics assay exploits the physiologically high endogenous expression of NKCC1 in immature neurons, which results in the depolarizing effects of GABA and can activate voltage-gated Ca 2+Therefore, in immature neurons, compounds that block NKCC1 are expected to inhibit Ca 2+ Response. Immature neurons were cultured for 3 days in vitro (3DIV) and loaded with a calcium-sensitive dye (Fluo4) for 15 minutes. Then, neurons were treated with bumetanide and furosemide (as positive controls) or each selected compound for 15 minutes. As a functional readout, fluorescence levels in these cultures were monitored before and after the application of GABA (100 μM for 20 seconds). To test neuronal viability at the end of the experiment, KC1 (90 mM for 40 seconds) was applied, which strongly depolarizes neurons, leading to a voltage-gated Ca 2+ To quantify the effects of bumetanide, furosemide, and selected compounds on NKCC1 inhibition, the fluorescence values ​​after GABA application were normalized to the fluorescence levels after KC1 application in treated neurons. Bumetanide, furosemide, and each of the selected compounds significantly reduced the fluorescence increase after GABA application compared to vehicle (DMSO)-treated controls. They did not affect the fluorescence levels after KC1 application (2a). The selected compounds showed a significant effect in inhibiting GABA-stimulated Ca 2+ The best efficacy was demonstrated in terms of response (2b), with fluorescence values ​​comparable to 10 μM bumetanide but even superior to 100 μM bumetanide, consistent with the chloride (YFP) assay.

[0574] Pharmacodynamic studies

[0575] The solubility of the selected NKCC1 inhibitor compound 3.17 in aqueous buffer and its stability in plasma and phase I metabolism in vitro (3a) were evaluated. The compound has high solubility (>250mM in PBS, pH 7.4) and is highly resistant to hydrolysis and phase I metabolism (t1 / 2>120min in plasma and t1 / 2>60min in liver microsomes). The data indicate that these compounds have good solubility and metabolic stability in vitro.

[0576] Cognitive impairment test

[0577] The efficacy of compound 3.17 in rescuing cognitive impairment was evaluated in four different cognitive tests in Ts65Dn mice ( Figure 4Adult Ts65Dn mice and their WT littermates (2 weeks old) were treated with 3.17 (ip 0.2 mg / kg) or its vehicle for one week.Over the next three weeks, the animals were tested in four different tasks to assess memory and cognition: a) novel object location task (Deidda, G. et al. Reversing excitatory GABAAR signaling restores synaptic plasticity and memory in a mouse model of Down syndrome. Nat Med 2015, 21(4), 318-26; Contestabile, A. et al. Lithium rescues synaptic plasticity and memory in Down syndrome mice. J Clin Invest 2013, 123(1), 348-61), b) novel object recognition test (Deidda G. 2015; Fernandez, F., Garner, CC, Object recognition memory is conserved in Ts1Cje, a mouse model of Down syndrome. Neuroscience letters 2007, 421, 137-141), c) T-maze task (Belichenko, NP et al. The "Down syndrome critical region" is sufficient in the mouse model to confer behavioral, neurophysiological, and synaptic phenotypes characteristic of Down syndrome. J Neurosci 2009, 29(18), 5938-48) (spontaneous change protocol, 11 trials) and d) fear conditioning test (Deidda G.2015; Costa, AC et al. Acute injections of the NMDA receptor antagonist memantine rescue performance deficits of the Ts65Dn mouse model of Down syndrome on a fearconditioning test. Neuropsychopharmacology 2008, 33(7), 1624-32). As expected, Ts65Dn mice treated with vehicle showed reduced performance compared to WT.Treatment with 3.17 improved cognitive performance in Ts65Dn mice (. Figure 4 ).

[0578] Example 3: NKCC1 and NKCC2 selectivity data

[0579] The compounds of the present invention were tested for selective inhibition of NKCC1 compared to NKCC2, as shown in Table 2 below.

[0580] Table 2

[0581]

[0582]

[0583]

[0584] Based on the example data reported in Table 2 above, some compounds showed better NKCC1 / NKCC2 selectivity.

[0585] As an advantage, the compounds do not have diuretic side effects.

[0586] In particular, compounds 1.7, 1.15, 2.2, 2.6, 2.7, 2.8, 3.8, 3.13, 3.14 and 3.17 have demonstrated the advantages described and are particularly preferred in the present invention.

[0587] In vitro thallium-based assay in HEK cells

[0588] The thallium-based assay is a standard assay for measuring the activity of potassium transporters, such as NKCC2, which is a sodium-potassium and chloride cotransporter. +) and then NaCl, they enter the cells via NKCC2, which is activated in the presence of chloride ions, and bind to a fluorescent dye, thereby determining an increase in fluorescence. This assay involves performing parallel testing in 96 wells, allowing for rapid and easy drug screening. Specifically, renal epithelial cells (HEK293) were transfected with either the NKCC2 transporter or a mock construct (control). Two days later, the cells were loaded with a thallium-sensitive fluorescent dye in Cl-free medium. After a one-hour incubation, the inhibitory activity of bumetanide and furosemide (as positive controls) and the novel compounds was tested by monitoring fluorescence after the application of thallium (simulating K) and then NaCl. Upon entering the cells via NKCC2 (activated by the presence of Cl), thallium binds to the fluorescent dye and increases fluorescence. After thallium application, NKCC2-transfected cells showed a robust increase in fluorescence levels compared to mock-transfected cells. Preincubation with bumetanide (10 μM) significantly reduced ion flux, thereby increasing fluorescence in NKCC2-transfected cells. A decrease in fluorescence was observed in mock-transfected cells treated with bumetanide and furosemide. This suggests that HEK293 cells express an endogenous transporter that is sensitive to bumetanide / furosemide. This latter result was used to normalize the fluorescence measurements obtained by this assay. Specifically, the ΔF / F0 values ​​of mock-transfected cells (control and treated) were subtracted from the corresponding ΔF / F0 values ​​of cells transfected with the Cl transporter. Using this assay, the ability of novel chemical entities to block NKCC2 was tested (results in Table 2).

[0589] Figure 17 Results of thallium assays are shown: a) Example traces obtained in a thallium-based assay on untransfected (mock) or NKCC2-transfected renal epithelial (HEK293) cells. Arrows indicate the addition of thallium (final concentration 2 mM) and NaCl stimulation (135 mM) to initiate the flux assay. b) Quantification of the effects of bumetanide, furosemide, and three example compounds (3.8, 3.13, 3.17) on NKCC2-transfected HEK293 cells in a thallium-based assay. Data represent mean ± sem from five independent experiments and are expressed as % of control. *P < 0.05, **P < 0.01, ***P < 0.001, Kruskal-Wallis ANOVA (Dunn's post hoc randomized controlled trial); ###P < 0.001, two-tailed unpaired Student's t-test.

[0590] VPA Autism Model

[0591] The efficacy of selected NKCC1 inhibitors was evaluated in vivo in a valproic acid (VPA)-induced autism mouse model to assess their ability to rescue altered social interactions. The VPA model was established by treating pregnant C57bl / 6j dams with 600 mg / kg (ip) VPA dissolved in PBS at gestational day 12.5. Offspring born to VPA-treated dams exhibit behaviors associated with core autism symptoms (Nicolini and Fahnestock, 2018). As a control, offspring from C57bl / 6j dams treated with PBS at gestational day 12.5 were used. To assess the efficacy of compounds in reverting social deficits, young male offspring from VPA- and PBS-treated dams were treated (ip) with 0.2 mg / kg compound 3.17 dissolved in PBS or 2% DMSO dissolved in PBS as a control for 7 days. Mice were then tested for social development and repetitive behaviors in various tests. Social development was assessed using the three-chamber test (Silverman et al., 2010). In the three-chamber test, mice were placed individually in a three-chamber box with an opening between the chambers. After ten minutes of free exploration, an unseen intruder was placed under a pencil cup in one chamber, and an empty pencil cup was placed in another chamber. The social index is the time that the animal explores the unseen intruder relative to the time the animal explores the pencil cup, and it is defined as: [(time spent with the intruder-time spent with the empty cup) / (time spent with the intruder+time spent with the empty cup)%]. In the second stage, a new intruder was rotated under the previous empty pencil box to measure the social novelty index, i.e., the time of exploring the new intruder compared to the object already encountered in the first 10 minutes. The social novelty index is measured as follows: [(time spent with the new intruder-time spent with the old intruder) / (time spent with the new intruder+time spent with the old intruder)%].

[0592] like Figure 18 As reported in Figure A, VPA mice treated with vehicle showed significantly lower sociability index and social novelty index compared to naive mice treated with vehicle. Treatment of VPA mice with compound 3.17 restored the sociability index and social novelty index to control levels.

[0593] Sociability during male-female interactions was then assessed (Drapeau et al., 2018). In this test, after 5 minutes of habituation, the mice were assessed for their approach to a female intruder mouse that had been placed in the same cage for 5 minutes. The time spent interacting was calculated as a measure of male-female social interaction. Figure 18As shown in Figure 3B, vehicle-treated VPA mice exhibited a significantly lower male-female interaction index than vehicle-treated naive mice. Treatment with compound 3.17 fully restored interaction in VPA mice. Finally, repetitive behaviors were assessed in two different tests. In the marble burying test (Eissa et al., 2018), mice were placed in a cage with 4 cm of litter and 15 (5*3) balls neatly placed on top. Repetitive behavior was assessed as the number of marbles buried in the litter. The grooming test is to assess grooming behavior, i.e. licking or scratching the head or other parts of the body with the front legs, which is a typical behavior of rodents (Campolongo et al., 2018). During the test, mice were placed in a cylindrical holder and repetitive grooming activity was measured within 5 minutes after 10 minutes of habituation. As Figure 18 As shown in Figures C and D, vehicle-treated VPA mice exhibited more repetitive behaviors (buried more marbles and spent more time grooming) than vehicle-treated naive mice. Treatment with compound 3.17 completely restored control levels of repetitive behaviors in VPA mice.

Claims

1. Use of a compound or a pharmaceutically acceptable salt thereof for preparing a medicament, wherein the compound is selected from the group consisting of: 3.17 3-(Dimethylsulfamoyl)-4-(8,8,8-trifluorooctylamino)benzoic acid, 5.7 3-morpholinosulfonyl-4-(8,8,8-trifluorooctylamino)benzoic acid, and 7.4 2-Hydroxy-5-sulfamoyl-4-(8,8,8-trifluorooctylamino)benzoic acid.

2. Use of a compound as defined in claim 1 for the preparation of a medicament for the treatment or prevention of a pathological condition associated with depolarizing GABAergic transmission, wherein the pathological condition is selected from the group comprising Down syndrome, neuropathic pain, stroke, cerebral ischemia, cerebral edema, hydrocephalus, traumatic brain injury, depressive-like behavior induced by brain trauma, autism spectrum disorder, fragile X, Rett syndrome, Asperger and DiGeorge syndromes, epilepsy, convulsions, West syndrome, glioma, anaplastic astrocytoma, Parkinson's disease, Huntington's disease, schizophrenia, anxiety, tuberous sclerosis and related behavioral problems, Dravet syndrome.

3. Use according to claim 2, wherein the pathological condition is selected from the group comprising autism, status epilepticus, glioblastoma.

4. A pharmaceutical composition comprising at least one compound or a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable excipient, and optionally one or more psychoactive substances and / or anti-inflammatory drugs; wherein the compound is selected from the group consisting of: 3.17 3-(Dimethylsulfamoyl)-4-(8,8,8-trifluorooctylamino)benzoic acid, 5.7 3-morpholinosulfonyl-4-(8,8,8-trifluorooctylamino)benzoic acid, and 7.4 2-Hydroxy-5-sulfamoyl-4-(8,8,8-trifluorooctylamino)benzoic acid.

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