Modulators of potassium channels and TRPV1 channels and uses thereof

Abstract

Formula I below: [Formula 1] The present invention provides compounds of formula Ia, Ib, III, IV or V as defined herein, and their use in modulating the activity of voltage-gated potassium channels and / or TRPV1 channels and treating conditions associated with the activity of these channels, such as conditions associated with neuronal vortex excitation.

Description

[Technical field]

[0001] Related Applications This application claims priority to U.S. Provisional Patent Application No. 63 / 300,293, filed January 18, 2022, the entire contents of which are incorporated herein by reference.

[0002] The present invention, in some embodiments thereof, relates to novel diphenylamine analogs, more particularly, but not limited to, newly designed diphenylamine analogs characterized by dual activity as modulators of both Kv7.2 / 3 and TRPV1 channels, which can be used to treat a variety of conditions associated with such channels, including, but not limited to, conditions associated with neuronal vortex excitation, such as tinnitus and pruritus. [Background technology]

[0003] Voltage-dependent potassium (Kv) channels allow potassium ions (K ) to pass through the cell membrane in response to changes in membrane voltage. + ) and can thereby modulate (increase or decrease) the electrical activity of the cell to regulate the excitability of the cell.

[0004] Functional Kv channels exist as multimeric structures formed in association with identical or different Kvα and / or Kvβ subunits. The α subunits contain six transmembrane domains, a pore-forming loop and a voltage sensor, and are arranged symmetrically around a central pore. The β subunits, i.e., auxiliary subunits, can interact with the α subunits and modify the properties of the channel complex, including, but not limited to, altering the electrophysiological or biophysical properties, expression levels or patterns of the channel.

[0005] Nine Kv channel α subunit families have been identified, designated Kv1 through Kv9. Thus, there is great diversity in Kv channel function resulting from the multiplicity of subfamilies, the formation of homomeric and heteromeric subunits within the subfamily, and additive effects associated with β subunits [MJ Christie, Clinical and Experimental Pharmacology and Physiology, 1995, 22 (12), 944-951].

[0006] The Kv7 channel family is composed of at least five members, including the following mammalian channels: Kv7.1, Kv7.2, Kv7.3, Kv7.4, Kv7.5, and one or more of their mammalian or non-mammalian equivalents or variants (including splice variants). The members of this family are alternatively referred to as KCNQ1, KCNQ2, KCNQ3, KCNQ4, and KCNQ5, respectively [Dalby-Brown et al., Current Topics in Medicinal Chemistry, 2006, 6, 999-1023].

[0007] The five members of this family have distinct expression patterns: Kv7.1 expression is restricted to the heart, peripheral epithelial and smooth muscle, whereas Kv7.2-Kv7.4 expression is restricted to the nervous system, including the hippocampus, cortical neurons and dorsal root ganglion neurons [for review, see e.g. Delmas P. & Brown D., Nature, 2005, 6, 850-862].

[0008] Neuronal Kv7 channels have been demonstrated to play an important role in the control of neuronal excitability. Kv7 channels, particularly the Kv7.2 / Kv7.3 heterodimer, underlie the M-current, a non-activating potassium current found in many neuronal cell types. This current has a characteristic time and voltage dependence that results in the stabilization of the membrane potential in response to multiple excitatory stimuli. Thus, the M-current is central in controlling neuronal excitability [for review, see e.g. Delmas.P & Brown.D, Nature, 2005, 6, 850-862].

[0009] Potassium channels are involved in many physiological processes, including regulation of heart rate, arterial dilation, insulin release, neuronal excitability, regulation of renal electrolyte transport, etc. Therefore, modulators of potassium channels are prime drug candidates, and research efforts are underway to develop new modulators as therapeutic agents.

[0010] Thus, given the important physiological role of Kv7 channels in the nervous system and the involvement of such channels in many diseases, the development of modulators of Kv7 channels is highly desirable.

[0011] Potassium channel modulators are divided into channel openers and channel blockers. A potassium channel opener that has attracted much attention is retigabine (N-(2-amino-4-(4-fluorobenzylamino)-phenyl)carbamic acid ethyl ester). Retigabine is highly selective for KCNQ2-5 type potassium channels. The use of retigabine for the treatment of neuropathic pain has been disclosed, for example, in U.S. Pat. No. 6,117,900 and European Patent Publication No. 1223927. Compounds related to retigabine have also been proposed for use as potassium channel modulators (see, for example, U.S. Pat. No. 6,472,165).

[0012] However, retigabine has been reported to have multiple actions in neuronal cells, including sodium and calcium channel blocking activity (Rundfeldt, C, 1995, Naunyn-Schmiederberg's Arch Pharmacol, 351 (Suppl): R160) and effects on GABA (gamma-aminobutyric acid) synthesis and transmission in rat neurons (Kapetanovic, IM, 1995, Epilepsy Research, 22, 167-173, Rundfeldt, C, 1995, Naunyn-Schmiederberg's Arch Pharmacol, 351 (Suppl): R160).

[0013] Other KCNQ potassium channel modulators are described, for example, in U.S. Patent Application Serial No. 10 / 075,521 (teaching 2,4-disubstituted pyrimidine-5-carboxamide derivatives as Kv7 modulators), U.S. Patent Application Serial No. 10 / 160,582 (teaching cinnamide derivatives as voltage-gated potassium channel modulators), U.S. Patent Nos. 5,565,483 and 10 / 312,123, 10 / 075,703 and 10 / 075,522 (teaching 3-substituted oxindole derivatives as voltage-gated potassium channel modulators), U.S. Patent No. 5,384,330 (teaching 1,2,4-triamino-benzene derivatives as potassium channel modulators), and U.S. Patent No. 6,593,349 (teaching bisarylamine derivatives as voltage-gated potassium channel modulators). U.S. Patent No. 6,291,442 teaches compounds containing two or three aromatic rings with a free carboxyl or carboxyls linked via an ester bond to a lower alkyl ester attached to one of the rings for modulating the Shaker class of voltage-gated potassium channels.

[0014] WO 2004 / 035037 and U.S. Patent Application Publication No. 20050250833 teach derivatives of N-phenylanthranilic acid and derivatives of 2-benzimidazolone, together with neuronal activity modulators, as potassium channel openers, in particular openers of voltage-gated potassium channels such as Kv7.2, Kv7.3 and KCv7.2 / 7.3 channels.

[0015] WO 2009 / 037707 teaches further derivatives of N-phenylanthranyl as potassium channel and / or TRPV1 modulators. An exemplary modulator disclosed in WO 2009 / 037707 is referred to as NH29.

[0016] [ka]

[0017] WO 2009 / 071947 and WO 2010 / 010380 teach derivatives of diphenylamine as potassium channel modulators. Exemplary modulators disclosed in these patent applications are designated NH34 and NH43.

[0018] [ka]

[0019] Transient receptor potential vanilloid type 1 (TRPV1) receptors are ligand-gated, nonselective cation channels that are activated by heat (usually >43°C), low pH (<6), and endogenous lipid molecules (e.g., anandamide, N-arachidonoyl-dopamine, N-acyl-dopamine, and products of lipoxygenase termed endovanilloids (e.g., 12- and 15-(S)-HPETE)). Apart from peripheral primary afferent neurons and dorsal root ganglia, TRPV1 receptors are expressed throughout the brain. Recent evidence indicates that TRPV1 receptor stimulation by endocannabinoids or capsaicin leads to analgesia, an effect associated with an increase in glutamate and activation of OFF cell populations in the rostral ventromedial medulla (RVM).

[0020] TRPV1 has also been shown to be involved in regulating body temperature, anxiety, and mediating long-term depression (LTD) in the hippocampus. TRPV1 channels are also present in sensory afferents that innervate the bladder. Inhibition of TRPV1 has been shown to improve symptoms of urinary incontinence.

[0021] TRPV1 modulators are described, for example, in WO 2007 / 054480, which teaches the effect of 2-(benzimidazol-1-yl)-acetamide derivatives in the treatment of TRPV1-related diseases. WO 2008 / 079683 teaches cyclohexyl and phenyl conjugated bicyclic compounds for inhibiting TRPV1 receptors. EP 01939173 teaches O-substituted dibenzyl urea or thiourea derivatives as TRPV1 receptor antagonists. WO 2008 / 076752 teaches benzimidazole compounds as potent TRPV1 modulators, and EP 01908753 teaches heterocyclidene acetamide derivatives as TRPV1 modulators.

[0022] The potassium channel Kv7.2 / 3 and the cation-nonselective channel TRPV1 transmit interval signals, have opposing functions, and are uniquely co-expressed in afferent peripheral sensory neurons (DRG sensory neurons). TRPV1 channels trigger pain signals, whereas Kv7.2 / 3 channels inhibit them. Compounds that simultaneously act as activators (e.g., openers) of Kv7.2 and inhibitors (e.g., blockers) of TRPV1 can inhibit vortex excitation of neurons associated with pathologies such as tinnitus and pruritus.

[0023] WO 2019 / 073471 discloses various modifications made to the structures taught in WO 2009 / 071947 and WO 2010 / 010380 that were found to exert dual modifications to both voltage-gated potassium channels and TRPV1. Two potential candidates disclosed in WO 2019 / 073471 are referred to herein as NH91 and NH101.

[0024] [ka] Summary of the Invention

[0025] According to an aspect of some embodiments of the present invention, a compound of formula I: [ka] (In the formula, A and B are each independently selected from aryl and heteroaryl; W is selected from -S-, -O-, -CR(OH)-, -C(=O), and NRx, where Rx together with one of the Ra substituents of ring A forms a nitrogen-containing heterocycle; n is an integer from 1 to 5; m is an integer from 0 to 4; Ra and Rb are each independently a substituent selected from alkyl, cycloalkyl, halo, haloalkyl, haloalkoxy, alkoxy, ether, aryl, heteroaryl, heteroalicyclic, aryloxy, hydroxy, amine, alkylamine, thiohydroxy, thioalkoxy, thioaryloxy, cyano, carboxylate, amide, carbamate, sulfonyl, and sulfonamide, or alternatively, at least two of the Ra substituents taken together form an alicyclic or heterocyclic ring, at least one of the Ra substituents taken together with Rx, if present, forms a nitrogen-containing heterocyclic ring, and / or at least two of the Rb substituents taken together form an alicyclic or heterocyclic ring, when n is greater than 1, each Ra is the same or different substituent, and when m is greater than 1, each Rb is the same or different substituent; R1, if present, is hydrogen or alkyl; V is -(CR2R3)kU; k is 0, 1 or 2; R2 and R3 are each independently selected from hydrogen, alkyl, cycloalkyl, and aryl; U is an amide (-C(=O)-NR 10 -) or its isostere, The Z is represented by the following formula II: [ka] (In the formula, u and q each independently represent an integer of 0 to 4 (provided that u+q is at least 2); X is selected from -O- and -NR9- or is absent; Y is OR 11 , S.R. 11 , amine (NR 12 R 13 ) or amide (-NR 12 -C(=O)-R 14 ) and the like, R5, R6, R7 and R8 are each independently selected from hydrogen, halo, alkyl, haloalkyl, cycloalkyl, heteroalicyclic, aryl, alkylamino, alkoxy, haloalkoxy, hydroxy, ether, and aryloxy, or alternatively, two of R5, R6, R7, R8 and R9 taken together form an alicyclic or heteroalicyclic ring; R9, R 10 , R 11 , R 12 , R 13 and R 14 are each independently selected from hydrogen, alkyl, cycloalkyl, and aryl, or alternatively, R, R, R, R, R, and R 11 Two of these, or R5, R6, R7, R8, R9, R 12 and R 13 Two of these together form an alicyclic or heteroalicyclic ring. A compound represented by the formula:

[0026] In some of the embodiments of any of the present applications, A and B are each aryl, for example, A and B are each phenyl.

[0027] In some of the embodiments of any of the compounds described herein, the compounds have formula Ia: [ka] (In the formula, W, V and Z are as defined in formula I; Ra1 to Ra5 ​​are each independently defined as Ra above, Each of Rb1 to Rb4 independently has the same definition as Rb above.)

[0028] In some of the embodiments described herein, m is other than 0 and at least one of the Rb substituents is halo.

[0029] In some of the embodiments described herein, m is 1.

[0030] In some of the embodiments described herein, the halo is para to the W.

[0031] In some of the embodiments described herein, Rb2 is halo, for example, fluoro.

[0032] In some of the embodiments of any of the embodiments described herein, B is a heteroaryl, for example, a nitrogen-containing heteroaryl, such as pyridine, pyrazole, pyrrole, imidazole, etc. In some embodiments, B is pyridine.

[0033] In some of the embodiments described herein, A is aryl, for example, phenyl.

[0034] In some of the embodiments of any of the present applications, the compound has formula Ib: [ka] (In the formula, W, V and Z are as defined in formula I; Ra1 to Ra5 ​​are each independently defined as Ra in the present application; Rb1, Rb2 and Rb4 each independently have the same definition as Rb in this application.)

[0035] In some of the embodiments described herein, n is 3, 4, or 5, and in some embodiments n is 3.

[0036] In some of the embodiments described herein, at least two of the R a substituents are selected from halo and alkoxy.

[0037] In some of any of the embodiments described herein, at least two of the R a substituents are each independently halo, for example chloro.

[0038] In some of the embodiments described herein, at least one of the R substituents is selected from alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamine, heteroaryl, and heteroalicyclic, or alternatively, two R substituents form a ring structure.

[0039] In some of the embodiments described herein, at least one of the R a substituents is alkyl or cycloalkyl.

[0040] In some of the embodiments described herein, at least one of the R a substituents is ortho to W.

[0041] In some of the embodiments described herein, at least one of the R a substituents is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamine, heteroaryl, or heteroalicyclic, and is ortho to W.

[0042] In some of the embodiments described herein, n is 3, one of the Ra substituents is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamine, heteroaryl, or heteroalicyclic and is ortho to W, and the other two of the Ra substituents are each halo.

[0043] In some of the embodiments described herein, n is 3; One of the Ra substituents is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamine, heteroaryl, or heteroalicyclic and is ortho to W, the other two of the Ra substituents are each halo, m is 1, and Rb is halo and is para to W.

[0044] In some of the embodiments of any of the present applications, Ra1 is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamine, heteroaryl, or heteroalicyclic.

[0045] In some of the embodiments described herein, Ra3 and Ra5 are each independently halo (eg, chloro).

[0046] In some of the embodiments described herein, X is absent.

[0047] In some of any of the embodiments described herein, at least one of R5, R6, R7, and R8 is each independently selected from alkyl, hydroxy, alkoxy, haloalkyl, ether, and halo, and / or at least two of R5, R6, R7, and R8 together form an alicyclic or heteroalicyclic ring.

[0048] In some of any of the embodiments described herein, at least two of R5, R6, R7, and R8 are each independently selected from alkyl, haloalkyl, and halo.

[0049] In some of any of the embodiments described herein, at least two of R5, R6, R7, and R8 are each independently alkyl.

[0050] In some of the embodiments described herein, u is 1 or 2.

[0051] In some of the embodiments described herein, R5 and R6 are each hydrogen.

[0052] In some of the embodiments described herein, q is 1 and at least one or each of R7 and R8 is alkyl.

[0053] In some of the embodiments described herein, q is 1 and at least one or each of R7 and R8 is hydrogen.

[0054] In some of any of the embodiments described herein, at least one of R5, R6, R7, and R8 is independently hydroxy, hydroxyalkyl, ether, heteroalicyclic (eg, oxygen-containing), or cycloalkyl.

[0055] In some of the embodiments of any of the embodiments described herein, the cycloalkyl is substituted with at least one hydrogen.

[0056] In some of any of the embodiments described herein, at least two of R5, R6, R7, and R8 taken together form an alicyclic ring.

[0057] In some of the embodiments described herein, X is absent, u is 1 or 2, q is 1, and R5, R6, R7 and R8 taken together form a cyclobutane.

[0058] In some of any of the embodiments described herein, at least two of R5, R6, R7, and R8 taken together form a heteroalicyclic ring, preferably a 4-, 5-, or 6-membered heteroalicyclic ring containing oxygen.

[0059] In some of the embodiments described herein, X is absent, u is 1 or 2, q is 1, and R5, R6, R7, and R8 taken together form a furan.

[0060] In some of the embodiments described herein, Y is OR 11 and R 11 is hydrogen.

[0061] In some of the embodiments described herein, Y is OR 11 and R 11 is alkyl, preferably alkyl substituted with at least one hydroxy, amido and / or carboxy.

[0062] In some of the embodiments described herein, Y is OR 11 and R11 is heteroaryl, or heteroalicyclic.

[0063] In some of the embodiments described herein, Y is NR 12 R 13 and R 12 forms a nitrogen-containing heteroalicyclic ring together with one or more of R5, R6, R7, and R8.

[0064] In some of the embodiments described herein, R 13 is hydrogen, hydroxy, alkyl, hydroxyalkyl or alkoxy.

[0065] In some of the embodiments described herein, Y is -NR 12 -C(=O)-R 14 and R 12 forms a nitrogen-containing heteroalicyclic ring together with one or more of R5, R6, R7, and R8.

[0066] In some of the embodiments of any of the present applications, Rx together with one of the Ra substituents forms a nitrogen-containing heterocycle.

[0067] In some of the embodiments of any of the present applications, Rx together with the Ra substituent ortho to W forms a nitrogen-containing heterocycle.

[0068] In some of the embodiments of any of the present applications, the compound has formula III: [ka] (In the formula, V and Z are as defined above in formula I; D is N or C-Rb3; Ra2 to Ra5, Rb1 to Rb4, V and Z are as defined in formula I, Ia or Ib herein; Rc1 and Rc2 are each independently as defined for Ra1 to Ra5 ​​in the present application.

[0069] In some of the embodiments of any of the present applications, Rc1 and Rc2 are each independently selected from hydrogen, alkyl, haloalkyl, and halo.

[0070] In some of the embodiments of any of the groups described herein, W is -S-.

[0071] In some of the embodiments of any of the present applications, the compound has formula IV: [ka] (In the formula, V and Z are as defined in formula I; D is N or C-Rb3; Ra1 to Ra5, Rb1 to Rb4, V and Z are as defined in formula I, Ia or Ib in this application.

[0072] In some of the embodiments described herein, Ra1 is selected from alkyl and cycloalkyl.

[0073] In some of the embodiments described herein, W is -CR1(OH).

[0074] In some of the embodiments of any of the present applications, the compound has formula V: [ka] (In the formula, V and Z are as defined in formula I herein; D is N or C-Rb3; Ra1 to Ra5, Rb1 to Rb4, V and Z are as defined in formula I, Ia or Ib in this application.

[0075] In some of the embodiments described herein, Ra1 is selected from alkyl and cycloalkyl.

[0076] In some of the embodiments described herein, W is -C(=O)-.

[0077] In some of the embodiments of any of the present applications, the compound has the formula VI: [ka] (In the formula, V and Z are as defined in formula I herein; D is N or C-Rb3; Ra1 to Ra5, Rb1 to Rb4, V and Z are as defined in formula I, Ia or Ib in this application.

[0078] In some of the embodiments described herein, Ra1 is selected from alkyl and cycloalkyl.

[0079] In some of the embodiments of any of the present disclosure, the compound is characterized by at least one of the following: a LogD value, determined as described herein, of greater than 4, greater than 5, or within the range of 4 and 7; a ligand lipophilicity efficiency (LLE) greater than 3, or greater than 5, as determined according to the present application; HLM Clint is less than 100 μl / min / mg as determined according to the present application; The kinetic solubility is greater than 20 micromolar, or greater than 30 micromolar.

[0080] In one aspect of some embodiments of the present invention, there is provided a pharmaceutical composition comprising a compound according to each embodiment and any combination thereof herein, and a pharma- ceutically acceptable carrier.

[0081] In one aspect of some embodiments of the present invention, there is provided a pharmaceutical composition comprising a compound according to any one of the embodiments and any combination thereof, or a compound according to any one of the embodiments and any combination thereof, for use in modulating the activity of a voltage-gated potassium channel.

[0082] In some of any of the embodiments described herein, the potassium channel is Kv7.2 / 7.3.

[0083] In some of any of the embodiments described herein, modulating comprises opening a channel.

[0084] In one aspect of some embodiments of the present invention, there is provided a pharmaceutical composition comprising a compound as described in each of the embodiments and any combination thereof herein, or a compound as described in each of the embodiments and any combination thereof herein, for use in modulating the activity of TRPV1.

[0085] In some of the embodiments of any of the present applications, modulating comprises inhibiting the activity of TRPV1.

[0086] In one aspect of some embodiments of the present invention, there is provided a pharmaceutical composition comprising a compound as described in any of the embodiments and any combination thereof herein, or a compound as described in any of the embodiments and any combination thereof herein, for use in modulating the activity of both voltage-gated potassium channels and TRPV1.

[0087] In some of any of the embodiments described herein, modulating the activity of a voltage-gated potassium channel comprises opening the channel, and modulating the activity of a TRPV1 channel comprises inhibiting the activity of the channel.

[0088] In some of any of the embodiments described herein, the potassium channel is Kv7.2 / 7.3.

[0089] In one aspect of some embodiments of the present invention, there is provided a pharmaceutical composition comprising a compound according to any of the embodiments and any combination thereof herein, or a compound according to any of the embodiments and any combination thereof herein, for use in treating a condition associated with activity of a voltage-gated potassium channel and / or a TRPV1 channel.

[0090] In some of any of the embodiments described herein, the condition is neuropathic pain, pruritus, or tinnitus.

[0091] Unless otherwise defined, all technical and / or scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this invention belongs.Although methods and materials similar or equivalent to those described herein can be used to practice or test embodiments of this invention, exemplary methods and / or materials are described below.In case of conflict, the patent specification, including definitions, shall prevail.In addition, the materials, methods, and examples are merely illustrative and are not necessarily intended to be limiting.

[0092] Certain embodiments of the present invention are described herein, by way of example only, with reference to the accompanying drawings, in which it is emphasized that the details shown hereinafter, with particular reference to the drawings, are for the purposes of illustration and for the purpose of detailed description of embodiments of the present invention. Similarly, from viewing the description together with the drawings, it will become apparent to those skilled in the art how embodiments of the present invention may be practiced. [Brief description of the drawings]

[0093] [Figure 1-1] FIG. 1 shows chemical structures of exemplary compounds within chemotype 1 according to some embodiments of the present invention. [Figure 1-2] Same as above [Figure 2-1] FIG. 2 shows chemical structures of exemplary compounds within chemotype 2 according to some embodiments of the present invention. [Figure 2-2] Same as above [Diagram 3]3A and 3B show chemical structures of exemplary compounds within chemotypes 3 and 4 (FIG. 3A) and potential enzyme-catalyzed conversions between them according to some embodiments of the present invention. [Figure 4] Figure 4A-B shows a homology model of human TRPV1 with reference (previously reported) compounds docked in the vanilloid pocket: reference compounds NH91 (000091, light yellow) and 000228 (e.g., a compound described in WO 2004 / 035037, green), the established TRPV1 agonist resiniferatoxin (RTX) serving as a reference molecule (Figure 4A), and a homology model of human Kv7.2 / 7.3 with previously reported 000091 (NH91) docked in the VSD (Figure 4B). A hydrophobic gasket and a hydrophilic channel are structural motifs common to both proteins, enabling the design of dual Kv7.2 / 7.3 and TRPV1 modulators. [Diagram 5] Figures 5A-B show representative illustrations of TRPV1 qSAR modeling. Figure 5A shows the overlay of exemplary effective TRPV1 inhibitors in some embodiments of the present invention. Figure 5B is a linear plot correlating experimental IC50 results (X-axis) with predicted qSAR IC50 values ​​for exemplary compounds in some embodiments of the present invention. [Figure 6-1] 6A-6C show data obtained on rat DRG neuron firing in response to activation by current injection (FIG. 6A), capsaicin (FIG. 6B) or both (FIG. 6C) with or without administration of various concentrations of AMG9810 (a TRPV1-specific antagonist), compound 273 (a Kv activator lacking TRPV1 inhibition), and exemplary compound 421-6 (an exemplary Kv and TRPV1 dual targeting compound), and retigabine (RET) as an exemplary Kv7.2 / 3 targeting compound with a similar EC50 as 421-6. [Figure 6-2] Same as above [Figure 6-3] Same as above [Figure 6-4]6D-E show dose-response curves showing both capsaicin-induced and current-induced neuronal excitation at various concentrations with compound 421-6 and inhibition by retigabine (FIGS. 6D and 6E, respectively). [Figure 7] 7A-C show exemplary data obtained for rat DRG neuron firing in response to current injection and capsaicin firing to 100 nM compound 552 (FIG. 7A), bar graphs showing comparative rat DRG firing responses between 100 nM each of compounds 421-6, 541, 552 and 533 (racemates), and exemplary data obtained from rat DRG neuron firing to current injection and capsaicin firing response in the presence of 100 nM compound 533 (racemic mixture). [Figure 8-1] FIG. 8A (Background Art) shows the experimental setup according to Zhang et al., 2005, EMBO J 24(24):4211-23, described in this application. [Figure 8-2] Figure 8B shows representative data obtained in assays of neurons pretreated with 100 ng / ml NGF for 4-5 days. NGF-naive rat DRGs were stimulated by repeated capsaicin administration in the presence or absence of 2 μM (top panel) or 100 nM (middle panel) 421-6. Following NGF pretreatment, tetanic bursts of firing were observed, showing reversible inhibition by 100 nM 421-6. [Figure 8-3] Figures 8C-D show an expanded view (Figure 8C) of the data shown in Figure 8B (bottom panel) and data obtained following NGF pretreatment in the presence of compound 421-6 (100 nM) and 2 μM compound 533 (Figure 8D). [Figure 9] Figures 9A-B show the inhibitory ability of compound 415 (5 μM, 1.5 min, Figure 9A) and previously reported compounds 291 (2 μM) and 414 (5 μM) (Figure 9B) on spontaneous neuronal activity in human-derived sensory nerves, demonstrating both the high potency of the compounds as well as their reversibility, with restored firing following washout of the compounds. [Figure 10-1]Figures 10A-D provide data showing the effect of compound 533 (533R) as a racemic mixture and its separated enantiomers 533p1 and 533p2. Figure 10A is a comparative plot showing activation of hKv7.2 / 3 measured using a high content fluorescent assay, comparing hKv7.2 / 3 activation by the 533 racemic mixture (533R) with its separated enantiomers 533p1 and 533p2, providing lower and higher activation potencies, respectively. Figure 10B provides that each of the 533 separated enantiomers modulates hTRPV1 in a different manner, with one (533p1) inhibiting hTRPV1 while the other (533p2) activates hTRPV1, and the 533 racemic mixture (533R) showing an average response. AMG9810, an antagonist of hTRPV1, served as a positive control. Figure 10C shows the EC / IC50 values ​​(in μM) of the separate enantiomers of compound 533 against the hKv7.2 / 3 and hTRPV1 targets (arrows indicate TRPV1 activation). Figure 10D shows data showing that 533p2 administration (100 nM) inhibits current-induced rat DRG spiking and enhances capsaicin-induced rat DRG spiking, consistent with the 533p2 TRPV1 activation properties (top panel), whereas administration of the 533 racemic mixture (533R) reduces current-induced spiking but achieves inhibition of capsaicin-induced spiking. [Figure 10-2] Same as above [Figure 11-1]11A-E are comparative plots showing caspain-induced inhibition of hTRPV1 activity measured using a fluorescent assay for compound 627 and AMG9810 as a positive control in cells stably expressing hTRPV1 (FIG. 11A) and cells expressing hKv7.2 / 3 and hTRPV1 (FIG. 11B). Comparative plots showing inhibition and activation of hKv7.2 / 3 activity measured using a fluorescent assay with compound 627 compared to a retigabine positive control (Figure 11C), activation of hKv7.2 / 3 activity measured using a fluorescent assay by very low concentrations (picomolar) of compound 627 (Figure 11D), and activation of hKv7.2 / 3 activity in electrophysiology by compound 627 measured by measuring whole cell currents of hKv7.2 / 3 expressed in CHO cells and clamping the membrane potential at -40 mV (1.5 seconds) (N=4) (from a holding potential of -90 mV) (Figure 11E). [Figure 11-2] Same as above [Figure 11-3] Same as above [Figure 12-1] Figures 12A-C show whole-cell current electrophysiology measurements (Figure 12A) in hTRPV1-expressing CHO cells (upper panel) and CHO cells co-expressing hKv7.2 / 3 and hTRPV1 upon activation (lower panel), dose-response curves (Figure 12B) showing the hTRPV1 inhibitory potential of CHO cells expressing hTRPV1 (right) and CHO cells that further co-express hKv7.2 / 3 and are activated (left), and exemplary capsaicin-induced currents in the presence of compound 627 in CHO cells co-expressing hTRPV1 and hKv7.2 / 3 but without hKv7.2 / 3 activation (Figure 12C). [Figure 12-2] Same as above [Figure 13]Figures 13A-B show exemplary capsaicin-induced currents in CHO cells co-expressing hTRPV1 and hKv7.2 / 3 in the presence of AMG9810, a known TRPV1 inhibitor (Figure 13A), and a dose-response comparison plot (Figure 13B) showing the hTRPV1 inhibitory potency in hTRPV1-expressing CHO cells and activated cells co-expressing hTRPV1 and hKv7.2 / 3 in the presence of 1 μM compound 627, AMG9810, and AMG9810 with retigabine (RET). [Figure 14] FIG. 14 depicts rat DRG membrane potential recordings showing habituation of action potentials to capsaicin challenge in the presence of 1 nM (top panel) and 0.1 nM (bottom panel) compound 627. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0094] The present invention, in some embodiments thereof, relates to novel diphenylamine analogs, and more particularly, but not limited to, newly designed diphenylamine derivative analogs characterized by dual activity as modulators of both potassium ion channels and TRPV1 channels, which can be used to treat various pathologies associated with such channels, such as, but not limited to, neuropathic pain.

[0095] Before describing at least one embodiment of the invention in detail, it is to be understood that the invention is not necessarily limited in its application to the details of construction and the arrangement of elements and / or methods set forth in the following description and / or illustrated in the drawings and / or examples. The invention is capable of other embodiments and of being practiced or carried out in various ways.

[0096] As described in the Examples section below, the inventors have conducted extensive research with the aim of finding analogs of diphenylamine derivatives that exhibit improved pharmaceutical profiles in terms of toxicity, bioavailability, solubility, and other pharmaceutical parameters and / or properties compared to known diphenylamine derivatives, and that exhibit improved therapeutic efficacy as dual modulators of both potassium ion channels (particularly Kv7.2 / 3 channels) and TRPV1 channels.

[0097] During the course of these intensive investigations, the inventors have found that certain structural modifications to the structures taught by WO 2009 / 071947 and WO 2019 / 073471 result in compounds that exhibit improved performance.

[0098] Small molecules have been designed and implemented that can be used to treat conditions in which dual modulation of potassium channel and TRPV1 activity is desirable. These small molecules can be particularly used to treat neuropathic pain, however, they can also be used to treat other conditions in which it is beneficial to modulate the activity of either or both potassium ion channels (particularly Kv7.2 / 3 channels) and TRPV1 channels, as described herein.

[0099] These compounds were designed based on a single compound approach to target multiple hyperexcitability-related mechanisms to achieve high efficacy and safety. The approach is based on targeting two cation channels, ligand-gated calcium and voltage-gated potassium channels, TRPV1 and Kv7.2 / 3, respectively, which are co-localized on sensory neurons and recognized as prominent regulators of neuronal excitability.

[0100] Small molecules were designed to test the effects of various modifications at multiple positions on previously discovered small molecules as described in Example 1. Classical and computational medicinal chemistry, as well as on-target and off-target screening methods, were used to identify potent, safe, and metabolically stable compounds for in-vivo testing. In addition, species selection studies were performed to aid in the discovery of appropriate animal models.

[0101] Using classical and computational medicinal chemistry, rational structure-activity relationship (SAR) studies resulted in the design and planning of nearly 500 compounds, of which 326 were screened for dual targeting properties. The results of these studies provided a substantial understanding of the SAR relationships. The primary goal was to replace the bridging aniline moiety, which was found to pose a significant toxicity risk. This resulted in the identification of four new chemotypes with potencies in the low μM range, and even lower in a heterologous system of CHO cells expressing ion channels.

[0102] The compounds found were assayed for off-target activity and cytotoxicity to predict a satisfactory safety profile. Potent compounds against both targets were tested for potential risks against Kv7.3 / 5, Kv7.4 and hERG, potential liver toxicity, TRPV1 polymodality uncoupling, and specificity for TRPA1. The compounds found exhibited improved tolerability profiles, mainly due to the complete loss of hERG inhibition, which occurred even at the maximum solubility of the compounds. In addition, some of the compounds found showed inhibition of capsaicin-induced activity of TRPV1, while not affecting pH- or heat-induced activation of TRPV1. The selective inhibition of capsaicin-gated TRPV1 currents, isolated from additional inhibitors against other TRPV1 activation modalities, may support the absence of risk of hyperthermia. This TRPV1 polymodality uncoupling was shown to be a good predictor for the absence of thermal dysregulation in preclinical and clinical settings.

[0103] The identified compounds were taken forward to in vitro DMPK studies to evaluate their potential druggability. LogD and human liver microsomes (HLM) were evaluated. Typically, the lower the LogD value of a compound (i.e., the more polar the compound), the lower the HLM value is expected. Indeed, for most compounds with LogD<4, HLM values ​​<100 (μL / min / mg) could be obtained. Other compounds showed favorable metabolic stability. A good correlation was observed between acceptable HLM and rat liver microsomes (RLM), suggesting the involvement of similar metabolic pathways in both species.

[0104] The use of species-specific constructs and species-specific cells demonstrated that no significant differences could be identified when comparing the inhibitory potency of test compounds whether of human, rat, or porcine origin. The lack of species differences in inhibitory potency against TRPV1 suggested that rodent models could be used to predict the efficacy of discovered compounds.

[0105] To study the inhibitory potential of the identified lead compounds against neuronal excitability in a physiologically relevant experimental system, primary neonatal rat DRG neurons, which contain normal compositions of neurons and glia, were used. The inhibitory potential of the identified compounds against both current-induced and capsaicin-induced neuronal activity was tested.

[0106] In the current simulations, the inhibitory potential of test compounds was shown to be mediated and identified through their effects on voltage-activated effectors such as Kv7.2 / 3 targets, whereas capsaicin-induced responses were identified as a contribution of capsaicin-gated TRPV1 targets, both to membrane depolarization upon activation and the resulting activation of downstream voltage-gated Kv7.2 / 3 targets.

[0107] Neuroexcitation is a physiological predictive readout, especially for pain amplification, and the additive effect of dual targeting of both TRPV1 and Kv7.2 / 3 in pain-related systems can be explored. For example, it was found that representative compound 421-6, which shows dual targeting properties of TRPV1 and Kv7.2 / 3 at 2 μM (EC50=2.1 μM for Kv7.2 / 3, IC50=1.8 μM for TRPV1), can dramatically reduce AP responses to both current activation and capsaicin activation. Compound 421-6 exhibited less than 100 nM inhibition of capsaicin-induced neuronal firing. This high inhibitory potency, which is significantly higher than the sum of the inhibition contributed by each target alone, indicates the synergistic effect of exemplary compound 421-6. The synergistic effect exerts a high inhibitory potency of the corresponding compounds and a high degree of specificity arising from the higher activity, which occurs only when both targets are co-expressed and their signaling crosses, the latter being unique to pain sensory nerves.

[0108] To provide additional physiological evidence in a relevant human system, human neural progenitor cells (hNPCs) differentiated into human sensory neurons were used. Using sudden and induced current application methods, it was shown that the test compounds have strong inhibitory potential on the excitability of human sensory neurons, with significant effects seen in rat DRG. The efficacy of the newly designed compounds was also shown to be superior to that of gabapentin, a standard treatment used clinically to treat neuropathic pain.

[0109] Thus, embodiments of the present invention relate to a novel family of compounds derived from diphenylamine derivatives that exhibit dual activity of opening potassium channels (e.g. Kv7.2 / 3) and inhibiting TRPV1 activity, and that exhibit an improved pharmacological profile compared to previously disclosed diphenylamine derivatives, and their use in the treatment of conditions associated with these channels, in particular pain, such as neuropathic pain.

[0110] Compound: The newly designed compounds according to this embodiment can be collectively represented by Formula I below: [ka] (In the formula, A and B are each independently selected from aryl and heteroaryl; W is selected from -S-, -O-, -CR(OH)-, -C(=O), and NRx, where Rx together with one of the Ra substituents of ring A forms a nitrogen-containing heterocycle; n is an integer from 1 to 5; m is an integer from 0 to 4; Ra and Rb are each independently a substituent selected from alkyl, cycloalkyl, halo, haloalkyl, haloalkoxy, alkoxy, ether, aryl, heteroaryl, heteroalicyclic, aryloxy, hydroxy, amine, alkylamine, thiohydroxy, thioalkoxy, thioaryloxy, cyano, carboxylate, amide, carbamate, sulfonyl, and sulfonamide, or alternatively, at least two of the Ra substituents taken together form an alicyclic or heterocyclic ring, at least one of the Ra substituents taken together with Rx, if present, forms a nitrogen-containing heterocyclic ring, and / or at least two of the Rb substituents taken together form an alicyclic or heterocyclic ring, when n is greater than 1, each Ra is the same or different substituent, and when m is greater than 1, each Rb is the same or different substituent; R1, if present, is hydrogen or alkyl; V is -(CR2R3)kU; k is 0, 1 or 2; R2 and R3 are each independently selected from hydrogen, alkyl, cycloalkyl, and aryl; U is an amide (-C(=O)-NR 10 -) or its isostere, The Z is represented by the following formula II: [ka] (In the formula, u and q each independently represent an integer of 0 to 4 (provided that u+q is at least 2); X is selected from -O- and -NR9- or is absent; Y is OR 11 , S.R. 11 , amine (NR 12 R 13 ) or amide (-NR 12 -C(=O)-R 14 ) and the like, R5, R6, R7 and R8 are each independently selected from hydrogen, halo, alkyl, haloalkyl, cycloalkyl, heteroalicyclic, aryl, alkylamino, alkoxy, haloalkoxy, hydroxy, ether, and aryloxy, or alternatively, two of R5, R6, R7, R8 and R9 taken together form an alicyclic or heteroalicyclic ring; R9, R 10 , R 11 , R 12 , R 13 and R 14 are each independently selected from hydrogen, alkyl, cycloalkyl, and aryl, or alternatively, R, R, R, R, R, and R 11 Two of these, or R5, R6, R7, R8, R9, R 12 and R 13 Two of these together form an alicyclic or heteroalicyclic ring.

[0111] As described in the Examples section below, in searching for compounds with improved pharmacological profiles, the inventors investigated various manipulations of the reported compounds. These manipulations were performed on various parts of the scaffold of the reported compounds, namely, rings A and B as shown in formula I, the side chains designated as -VZ in formula I, particularly Z, and the bridge between rings A and B, designated as W in formula I. The inventors found that the best performing compounds were those in which the amine bridge of the reported compounds was replaced with another moiety, defined herein as W in formula I.

[0112] The A and B rings, the -VZ side chain, and the W bridge in various embodiments of the compounds of formula I described herein are described below.

[0113] Ring A and Ring B: In some of the embodiments described herein, A and B are each aryl, as defined herein. In certain of these embodiments, A and B are each phenyl; such compounds are collectively designated by formula Ia.

[0114] [ka] (In the formula, W, V and Z are as defined in formula I; Ra1 to Ra5 ​​each independently have the same definition as Ra, and represent a substituent of ring A; Rb1 to Rb4 each independently have the same definition as Rb above and represent a substituent of ring B.

[0115] In some of the embodiments of any of the embodiments described herein (e.g., Formula I, Ia, or other related formulas), m is other than 0 and is 1, 2, 3, or 4, and at least one of the Rb substituents is halo. Other substituents, when present (e.g., when m is 2, 3, or 4), are as defined herein for Rb.

[0116] In some of the embodiments of any of the embodiments described herein (e.g., Formula I, Ia, or other related formulas), m is 1. In some of these embodiments, the Rb substituent is halo.

[0117] In some of any of the embodiments described herein, m is 1 and the Rb substituent is para to W, i.e., Rb2 in formula Ia is a substituent and the other Rb substituents, e.g., Rb1, Rb3 and Rb4, are each hydrogen. In some of these embodiments, Rb in formula I or Rb2 in formula Ia is halo.

[0118] In some of the embodiments described herein, when Rb or Rb2 is halo, the halo is fluoro.

[0119] In some of the embodiments described herein, for formula I, B is heteroaryl. A can be aryl or heteroaryl, and is preferably aryl, e.g., phenyl.

[0120] In some of these embodiments, B is a nitrogen-containing heteroaryl as described herein, which may, for example, have 1, 2 or 3 nitrogen atoms in the ring, and in some embodiments, B is pyridine.

[0121] In some of the embodiments of any of the invention described herein, in formula I, B is heteroaryl, eg, optionally having 1, 2 or 3 nitrogen atoms in the ring, and A is aryl, eg, phenyl.

[0122] In some of the embodiments of any of the present applications, in formula I, B is pyridine and A is aryl, for example, phenyl.

[0123] In some of the embodiments of any of the compounds described herein, in formula I, B is pyridine and A is phenyl; such compounds are collectively designated by formula Ib.

[0124] [ka] (In the formula, W, V and Z are as defined in formula I; Ra1 to Ra5 ​​are each independently defined as Ra in the present application; Rb1, Rb2 and Rb4 each independently have the same definition as Rb in this application.)

[0125] In some of any of the embodiments described herein for formulae I, Ia, Ib and any other related formulae described herein, n is at least 3, and can be 3, 4, or 5, such that at least three positions on ring A are substituted (with Ra substituents other than hydrogen), or at least three of Ra1-Ra5 in formula Ia or Ib are other than hydrogen.

[0126] In some of any of the embodiments described herein for formulae I, Ia, Ib and any other related formulae described herein, n is 3, and thus, three positions of ring A are substituted (by Ra substituents other than hydrogen), or three of Ra1-Ra5 in formula Ia or Ib are other than hydrogen.

[0127] In some of any of the embodiments described herein for Formula I, Ia, Ib, and any other related formulas described herein, two or more of the Ra substituents (e.g., at least two of Ra1-Ra5 in Formula Ia or Ib) are selected from halo and alkoxy.

[0128] In some of these embodiments, two of the Ra substituents (e.g., two of Ra1-Ra5 in formula Ia or Ib) are each independently alkoxy (which may be the same or different) and two of the Ra substituents (e.g., two of Ra1-Ra5 in formula Ia or Ib) are each independently halo (which may be the same or different), or one of the Ra substituents (e.g., one of Ra1-Ra5 in formula Ia or Ib) is alkoxy and one of the Ra substituents (e.g., one of Ra1-Ra5 in formula Ia or Ib) is halo.

[0129] In some of any of the embodiments described herein, when one or more of the Ra substituents (e.g., one of Ra1-Ra5 in formula Ia or Ib) is alkoxy, it is a lower alkoxy of 1 to 4 carbon atoms in length, such as methoxy, ethoxy, propoxy, isopropoxy, amyloxy, butoxy and / or isobutoxy, preferably methoxy.

[0130] In some of these embodiments, two of the Ra substituents (eg, two of Ra1-Ra5 in formula Ia or Ib) are each independently halo, for example chloro.

[0131] In some of any of the embodiments described herein of formulae I, Ia, Ib, and any other related formulae described herein, at least one of the Ra substituents (e.g., one of Ra1-Ra5 in formula Ia or Ib) is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamine, heteroaryl, or heteroalicyclic; or, alternatively, two of the Ra substituents (e.g., two of Ra1-Ra5 in formula Ia or Ib) taken together form a group.

[0132] In some of any of the embodiments described herein of Formula I, Ia, Ib, and any other related formulas described herein, at least one of the Ra substituents (e.g., one of Ra1-Ra5 in Formula Ia or Ib) is alkyl, haloalkyl, cycloalkyl, and / or aryl.

[0133] In some of any of the embodiments described herein of Formula I, Ia, Ib, and any other related formulas described herein, at least one of the Ra substituents (e.g., one of Ra1-Ra5 in Formula Ia or Ib) is alkyl or cycloalkyl.

[0134] In some of the embodiments of any of formulae I, Ia, Ib and any other related formulae described herein, the Ra substituent is ortho to W, i.e., Ra1 in formula Ia or Ib is other than hydrogen.

[0135] In some of these embodiments, the Ra substituent is ortho to W, i.e., Ra1 in formula Ia or Ib is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamine, heteroaryl, or heteroalicyclic, as described in the corresponding embodiments herein, preferably alkyl or cycloalkyl.

[0136] In some of the embodiments of any of the present invention according to Formula I, Ia, Ib, and any other related formulas described herein, one or more of the Ra substituents (e.g., Ra1 to R in Formula Ia or Ib) may be When one of a5) is an alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamine, heteroaryl, or heteroalicyclic, as described in the corresponding embodiments herein, at least one such Ra substituent is ortho to W in formula I.

[0137] In some of the embodiments of any of the formulae I, Ia, Ib, and any other related formulae described herein, n is 3, one of the Ra substituents is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamine, heteroaryl, or heteroalicyclic and is ortho to W (i.e., Ra1 in formula Ia or Ib), and the other two Ra substituents are each halo.

[0138] In some of the embodiments of any of the present invention according to formulae Ia, Ib and any other related formulae described herein, n is 3, Ra1 is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamine, heteroaryl or heteroalicyclic, preferably alkyl or cycloalkyl, and the other two Ra substituents (Ra2, Ra3, Ra4 and Ra5) are each halo, preferably chloro. In some of these embodiments, Ra3 and Ra5 are each halo, preferably chloro.

[0139] In some of the embodiments of any of the present applications according to formula I, n is 3, one of the Ra substituents is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamine, heteroaryl or heteroalicyclic, preferably alkyl or cycloalkyl, and is ortho to W, the other two Ra substituents are each halo, preferably chloro, m is 1, and Rb is halo, preferably chloro, and is para to W.

[0140] In some of the embodiments of any of the present applications according to formula Ia or Ib, Ra1 is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamine, heteroaryl, or heteroalicyclic.

[0141] In some of any of the embodiments described herein according to formula Ia or Ib, Ra3 and Ra5 are each independently halo (eg, chloro).

[0142] -VZ side chain: In some of the embodiments described herein, V is -(CR2R3)kU, k is 0, 1, or 2, R2 and R3 are each independently hydrogen, alkyl, cycloalkyl, and aryl, and U is an amide (-C(=O)-NR 10 -) or its isomer.

[0143] In some of the embodiments described herein, k is 1.

[0144] In some of the embodiments described herein, k is 1 and R2 and R3 are each hydrogen.

[0145] In an exemplary embodiment, U is an amide (-C(=O)-NR 10 -), and in some of these embodiments, R 10 is hydrogen.

[0146] In an alternative embodiment, U is an amide isostere.

[0147] By "isostere" is meant a moiety that has similar shape and / or electronic properties and / or biological activity as the corresponding parent moiety (here an amide) and is therefore expected to be recognized by the body in the same way as the parent moiety. In the context of pharmacological chemistry, isosteres are also called bioisosteres.

[0148] Examples of isosteres of amides include, but are not limited to, sulfonamides, phosphonamidates, thioamides, esters, ureas, fluoroalkenes, alkenes, tetrazoles, oxadiazoles, thiazoles, triazoles, imidazoles, pyrazoles, indoles, pyridines, pyrazines, carbamates, amidines, trifluoroethylamines, hydroxyethyls, alpha-difluoroketones, and oxetanylamines.

[0149] Exemplary isosteres are shown in FIG. 1 and are compound 526, an oxetanylamine, and compound 527, a triazole.

[0150] In some of the embodiments of any of Formulas I, Ia, Ib, and any other related formulas described herein, k is 1, R2 and R3 are each hydrogen, U is an amide (-C(=O)-NR10-) or an isostere thereof as described herein, and Z is represented by Formula II.

[0151] The moiety Z preferably contains one or more oxygen atoms, for example as part of the backbone of a side chain (e.g. when X is O or when Y is OR 11 and R 11 is other than hydrogen), as part of the terminal group of the side chain (e.g., Y is OR 11 and R 11 is hydrogen), and / or as a side chain backbone substituent (e.g., when one or more of R5, R6, R7, and R8 is an oxygen-containing heteroalicyclic ring (e.g., furan or oxetane), or when two or more of R5, R6, R7, and R8 together form an oxygen-containing heteroalicyclic ring, and / or when one or more of R5, R6, R7, and R8 is hydroxy or alkoxy or an oxygen-containing heteroalicyclic ring as described herein, or hydroxy-substituted cycloalkyl). Combinations of any of the foregoing are contemplated.

[0152] In some of any of the embodiments described herein for Formula I, Ia, Ib, and any other related formulas, X is absent.

[0153] In some of the embodiments of any of Formulas I, Ia, Ib and any other related formulas described herein, at least one of R5, R6, R7 and R8 is independently selected from alkyl, hydroxy, alkoxy, haloalkyl, ether, and halo, and / or at least two of R5, R6, R7 and R8 together form an alicyclic or heteroalicyclic ring.

[0154] In some of any of the embodiments described herein for Formula I, Ia, Ib, and any other related formulas, two or more of R5, R6, R7, and R8 are independently selected from alkyl, haloalkyl, and halo, and in some embodiments, two or more of R5, R6, R7, and R8 are alkyl.

[0155] In some of the embodiments of Formula I, Ia, Ib, and any other related formulas described herein, u is 1 or 2. It is noted that when u is greater than 1 and is comprised of more than one (CR5R6) moiety, R5 and R6 can each be the same or different moieties.

[0156] In some of any of the embodiments described herein for Formula I, Ia, Ib, and any other related formulas, R5 and R6 are each hydrogen.

[0157] In some of any of the embodiments described herein for Formula I, Ia, Ib, and any other related formulas, u is 1 or 2, and R5 and R6 are each hydrogen.

[0158] In some of the embodiments of any of Formulas I, Ia, Ib, and any other related formulas described herein, q is 1 or 2. It is noted that when q is greater than 1 and is comprised of two or more (CR7R8) moieties, R7 and R8 can each be the same or different moieties.

[0159] In some of any of the embodiments of Formula I, Ia, Ib, and any other related formulas described herein, q is 1 and at least one or each of R7 and R8 is alkyl.

[0160] In some of the embodiments of any of Formulas I, Ia, Ib and any other related formulas described herein, u is 1 or 2, R5 and R6 are each hydrogen, q is 1, and at least one or each of R7 and R8 is alkyl. Exemplary such compounds include compounds 417, 417-4, 532, 270, 519, 421-6, 526, 527 shown in Figures 1, 2, and 3A.

[0161] In some of any of the embodiments of Formula I, Ia, Ib, and any other related formulas described herein, q is 1 and at least one or each of R7 and R8 is hydrogen.

[0162] In some of the embodiments of any of formulas I, Ia, Ib, and any other related formulas described herein, u is 1 or 2, R5 and R6 are each hydrogen, q is 1, and at least one or each of R7 and R8 is hydrogen. Exemplary such compounds include compounds 552 and 627 shown in FIG.

[0163] In some of the embodiments of any of the present invention according to formulae I, Ia, Ib, and any other related formulae described herein, one or more of R5, R6, R7, and R8 are each independently hydroxy, hydroxyalkyl, ether, heteroalicyclic (e.g., oxygen-containing, as described herein), or cycloalkyl (e.g., 3, 4, or 5 atoms). In some of these embodiments, the cycloalkyl is substituted with one or more hydroxy groups. In some of these embodiments, u is 1 or 2. In some of these embodiments, q is 1. In some of these embodiments, u is 1 or 2 and q is 1. Exemplary such compounds include compounds 486, 490, 533, 536, 569, 573, 482, 513, 514, 414, 415, and 482 shown in Figures 1, 2, and 3A.

[0164] In some of the embodiments of any of the present invention according to formulae I, Ia, Ib and any other related formulae described herein, two or more of R5, R6, R7 and R8 together form an alicyclic ring, e.g., a cyclobutane. In some of these embodiments, u is 1 or 2. In some of these embodiments, q is 1. In some of these embodiments, u is 1 or 2 and q is 1. Exemplary such compounds include compounds 498, 499, 500, 501, 502, 512, 480, 481, 483, 484, 485, 489, 488, 534, 535 shown in Figures 1, 2 and 3A.

[0165] In some of the embodiments of any of the present invention according to formulae I, Ia, Ib and any other related formulae described herein, two or more of R5, R6, R7 and R8 together form a heteroalicyclic ring, preferably an oxygen-containing 4-, 5- or 6-membered heteroalicyclic ring. In exemplary embodiments, the oxygen-containing heteroalicyclic ring is a furan. In some of these embodiments, u is 1 or 2. In some of these embodiments, q is 1. In some of these embodiments, u is 1 or 2 and q is 1. In some of these embodiments, X is absent. Exemplary such compounds include compounds 503, 504, 505, 506, 507, 537 shown in Figure 1 and Figure 3A.

[0166] In any of the embodiments described herein for Formula I, Ia, Ib, and any other related formulas, in combination with any of the embodiments described herein for Formula II, Y is OR 11 and R 11 is hydrogen.

[0167] In any of the embodiments described herein for Formula I, Ia, Ib, and any other related formulas, in combination with any of the embodiments described herein for Formula II, Y is OR 11 and R 11 is alkyl, preferably substituted with at least one hydroxy, amido and / or carboxy. Exemplary such compounds include compounds 486, 483 and 488 shown in Figures 1 and 2.

[0168] In any of the embodiments described herein for Formula I, Ia, Ib, and any other related formulas, in combination with any of the embodiments described herein for Formula II, Y is OR 11 and R 11 is heteroaryl or heteroalicyclic as defined herein. An exemplary such compound is compound 481 (see FIG. 2).

[0169] In some of the embodiments of any of the formulas I, Ia, Ib, and any other related formulas described herein, in combination with any of the embodiments described herein for formula II, Y is NR 12 R 13 It is.

[0170] In some of the embodiments of any of the formulas I, Ia, Ib, and any other related formulas described herein, in combination with any of the embodiments described herein for formula II, Y is NR 12 R 13 and R 12 R, R, R, and R taken together with one or more of R, R, R, and R form a nitrogen-containing heteroalicyclic ring (e.g., azetidine). In some of these embodiments, R 13 is hydrogen, hydroxy, alkyl, hydroxyalkyl, alkylene glycol, or alkoxy. Exemplary such compounds include compounds 487 and 491 shown in FIG.

[0171] In some of the embodiments of any of the formulas I, Ia, Ib, and any other related formulas described herein, in combination with any of the embodiments described herein for formula II, Y is -NR 12 -C(=O)-R 14 It is.

[0172] In some of the embodiments of any of the formulas I, Ia, Ib, and any other related formulas described herein, in combination with any of the embodiments described herein for formula II, Y is -NR 12 -C(=O)-R 14 and R 12 R5, R6, R7, and R8 taken together with one or more of them form a nitrogen-containing heteroalicyclic ring (e.g., azetidine). Exemplary such compounds include compound 492 shown in FIG.

[0173] W crosslinking: In some of the embodiments of any of the present invention according to formula I, Ia, Ib and II, and any combination thereof, W is -NRx-, and Rx together with one of the Ra substituents forms a nitrogen-containing heterocycle. The nitrogen-containing heterocycle can be a 4-, 5-, 6-, 7-, or 8-membered ring, preferably a 4-, 5-, or 6-membered ring. The heterocycle can be a heteroaryl or heteroalicyclic ring fused to ring A.

[0174] In some of these embodiments, Rx forms a heterocycle with the Ra substituent ortho to W (e.g., Ra1 of formula Ia or Ib), as described herein. Such compounds are also referred to herein as chemotype I compounds.

[0175] In some corresponding embodiments of Formula I, A is aryl (e.g., phenyl) and B is aryl (e.g., phenyl) or heteroaryl. In some embodiments, when B is heteroaryl, the heteroaryl is a nitrogen-containing heteroaryl and may contain 1, 2, or 3 nitrogen atoms in the ring. In some embodiments, when B is heteroaryl, the heteroaryl is pyridine.

[0176] In some of the embodiments of any of the present applications, for compounds of chemotype I, the compounds are collectively represented by formula III.

[0177] [ka] (In the formula, -VZ (side chain moiety) is as described herein for any corresponding embodiment or any combination thereof; D is N or C-Rb3; Ra2-Ra5, Rb1-Rb4, V and Z are as defined and described in formula I, Ia or Ib herein for any corresponding embodiment or any combination thereof; Rc1 and Rc2 are each independently a substituent as defined in this application as Ra1 to Ra5 ​​for any corresponding embodiment or any combination thereof.

[0178] In these embodiments, Rx and Ra1 together form a nitrogen-containing heteroaryl fused to ring A.

[0179] In some of the embodiments of any of the formulas described herein for III, Rc1 and Rc2 can each independently be hydrogen or a substituent (alkyl, haloalkyl, and / or halo). In exemplary embodiments, Rc1 is hydrogen and Rc2 is alkyl, preferably lower alkyl having a length of 1 to 4 carbon atoms, such as methyl, or haloalkyl, such as trihaloalkyl (such as CF3), or halo, such as fluoro or chloro.

[0180] Exemplary compounds of Formula III, chemotype I, are shown in FIG.

[0181] In some of any of the embodiments described herein according to Formula I, Ia, Ib, and II, and any combination thereof, W is -S-. Such compounds are also referred to herein as chemotype II compounds.

[0182] In some corresponding embodiments of Formula I, A is aryl (eg, phenyl) and B is aryl (eg, phenyl) or heteroaryl, such as a nitrogen-containing heteroaryl described herein (eg, pyridine).

[0183] In some of the embodiments of any of the chemotype II compounds described herein, the compounds are collectively represented by formula IV.

[0184] [ka] (In the formula, -VZ (side chain moiety) is as described herein for any corresponding embodiment or any combination thereof; D is N or C-Rb3; Ra1 to Ra5, Rb1 to Rb4, V and Z are as defined in formula I, Ia or Ib in any of the corresponding embodiments or combinations thereof.

[0185] In an exemplary embodiment, Ra1 is alkyl or cycloalkyl as described herein for Formula I, Ia, or Ib.

[0186] Exemplary compounds of formula IV, chemotype II, are shown in FIG.

[0187] In some of the embodiments of any of the formulae I, Ia, Ib, and II, and any combination thereof, described herein, W is -CR1(OH)- and R1 is as defined herein. Such compounds are also referred to herein as chemotype III compounds.

[0188] In some corresponding embodiments of Formula I, A is aryl (eg, phenyl) and B is aryl (eg, phenyl) or heteroaryl, such as a nitrogen-containing heteroaryl described herein (eg, pyridine).

[0189] In some of the embodiments of any of the chemotype III compounds described herein, the compounds are collectively represented by formula V:

[0190] [ka] (In the formula, -VZ (side chain moiety) is as described herein for any corresponding embodiment or any combination thereof; D is N or C-Rb3; Ra1 to Ra5, Rb1 to Rb4, V and Z are as defined in formula I, Ia or Ib in any of the corresponding embodiments or combinations thereof.

[0191] In an exemplary embodiment, Ra1 is alkyl or cycloalkyl, as defined herein for Formula I, Ia, or Ib.

[0192] In some of any of the embodiments described herein according to Formula I, Ia, Ib, and II, and any combination thereof, W is -C(=O)-. Such compounds are also referred to herein as chemotype IV compounds.

[0193] In some corresponding embodiments of Formula I, A is aryl (eg, phenyl) and B is aryl (eg, phenyl) or heteroaryl (eg, pyridine).

[0194] In some of the embodiments of any of the chemotype IV compounds described herein, the compounds are collectively represented by formula VI.

[0195] [ka] (In the formula, -VZ (side chain moiety) is as described herein for any corresponding embodiment or any combination thereof; D is N or C-Rb3; Ra1 to Ra5, Rb1 to Rb4, V and Z are as defined in formula I, Ia or Ib in this application.

[0196] In an exemplary embodiment, Ra1 is alkyl or cycloalkyl, as described for Formula I, Ia, or Ib.

[0197] Exemplary compounds of formulas V and VI, chemotypes III and IV, are shown in Figure 3A. As described in the Examples section below and shown in Figure 3B, chemotype III compounds can be enzymatically converted to chemotype IV compounds, and vice versa.

[0198] In some of the embodiments of any of the inventions described herein, the newly designed compounds exhibit one or more, or two or more, or three or more, and preferably all, of the following characteristics: a LogD value, determined as described herein, of greater than 4, greater than 5, or within the range of 4 and 7; a ligand lipophilicity efficiency (LLE) greater than 3, or greater than 5, as determined according to the present application; HLM Clint is less than 100 μl / min / mg as determined according to the present application; The kinetic solubility is greater than 20 micromolar, or greater than 30 micromolar.

[0199] With respect to any of the embodiments described herein and any combination thereof, the compound may be in the form of a salt, for example a pharma- ceutically acceptable salt.

[0200] As used herein, the phrase "pharmaceutically acceptable salt" refers to a charged species of the parent compound and its counterion, which is typically used to adjust the solubility properties of the parent compound and / or reduce any significant irritation of the parent compound to an organism, but does not abolish the biological activity or properties of the administered compound. Pharmaceutically acceptable salts of the compounds described herein can also be formed during the synthesis of the compounds, for example, during the isolation of the compound from a reaction mixture or the process of recrystallizing the compound.

[0201] In some circumstances of this embodiment, the pharma- ceutically acceptable salts of the compounds described herein may be acid addition salts that, if desired, comprise at least one basic group of the compound (e.g., an amine and / or amide and / or nitrogen atom in a heterocyclic group) in a positively charged state (e.g., the basic group is protonated) in combination with at least one counterion derived from a selected acid to form a pharma- ceutically acceptable salt.

[0202] Thus, the acid addition salts of the compounds described herein can be a complex formed between one or more basic groups of the compound and one or more equivalents of an acid.

[0203] Depending on the stoichiometric ratio between the charged groups in the compound and the counterions in the salt, acid addition salts can be mono- or polyaddition salts.

[0204] As used herein, the phrase "monoaddition salt" refers to a salt with a 1:1 stoichiometric ratio of counterion to charged state of compound, where the addition salt contains one molar equivalent of counterion per molar equivalent of compound.

[0205] As used herein, the phrase "polyaddition salt" refers to a salt in which the stoichiometric ratio of counterion to charged state of compound is greater than 1:1, e.g., 2:1, 3:1, 4:1 or higher, the addition salt containing more than one molar equivalent of counterion per molar equivalent of compound.

[0206] Examples of pharma- ceutically acceptable salts include, but are not limited to, acid addition salts thereof with ammonium or guanidinium cations.

[0207] Acid addition salts can include a variety of organic and inorganic acids, including, but not limited to, hydrochloric acid resulting in hydrochloric acid addition salts, hydrobromic acid resulting in hydrobromic acid addition salts, acetic acid resulting in acetic acid addition salts, ascorbic acid resulting in ascorbic acid addition salts, benzenesulfonic acid resulting in besylate addition salts, camphorsulfonic acid resulting in camphorsulfonic acid addition salts, citric acid resulting in citric acid addition salts, maleic acid resulting in maleic acid addition salts, malic acid resulting in malic acid addition salts, methanesulfonic acid resulting in methanesulfonic acid (mesylate) addition salts, naphthalenesulfonic acid resulting in naphthalenesulfonic acid addition salts, oxalic acid resulting in oxalic acid addition salts, phosphoric acid resulting in phosphoric acid addition salts, toluenesulfonic acid resulting in p-toluenesulfonic acid addition salts, succinic acid resulting in succinic acid addition salts, sulfuric acid resulting in sulfuric acid addition salts, tartaric acid resulting in tartrate addition salts, and trifluoroacetic acid resulting in trifluoroacetic acid addition salts. Each of these acid addition salts can be mono- or polyaddition salts as these terms are defined herein.

[0208] The present embodiments further include any enantiomers, diastereomers, prodrugs, solvates, hydrates, and / or pharma- ceutically acceptable salts of the compounds described herein.

[0209] The term "enantiomer" as used herein means a stereoisomer of a compound that can be superimposed on its counterpart only by complete inversion / reflection (mirror image) of each other. Enantiomers are said to have a "handedness" and are referred to each other as right-handed and left-handed. Enantiomers have identical chemical and physical properties except when present in an environment that itself has a handedness, such as all living systems. In the context of this embodiment, a compound may exhibit one or more chiral centers, each of which may exhibit an R or S configuration and any combination, and compounds according to some embodiments of the present invention may have any chiral center that exhibits an R or S configuration.

[0210] The term "diastereomers" as used herein refers to stereoisomers that are not enantiomers of each other. Diastereomers occur when two or more stereoisomers of a compound have different configurations at one or more (but not all) of the equivalent (related) stereocenters, but are not mirror images of each other. If two diastereomers differ from each other at only one stereocenter, they are epimers. Each stereocenter (chiral center) gives rise to two different configurations, and thus two different stereoisomers. In the context of the present invention, embodiments of the present invention encompass compounds with multiple chiral centers occurring in any combination of configurations, i.e., any diastereomers.

[0211] The term "prodrug" refers to an agent that is converted into an active compound (the active parent drug) in vivo. Prodrugs are usually useful for facilitating administration of the parent drug. Prodrugs may, for example, be bioavailable by oral administration whereas the parent drug is not. Prodrugs may also have improved solubility compared to the parent drug in pharmaceutical compositions. Prodrugs are often used to obtain a sustained release of an active compound in vivo. Exemplary prodrugs of the compounds according to some of the embodiments of the present application include esters of hydroxy groups (e.g., in the side chain), including carboxylate esters, phosphate esters, and the like.

[0212] The term "solvate" refers to a complex of variable stoichiometry (e.g., di-, tri-, tetra-, penta-, hexa-, etc.) formed by a solute (a compound of the invention) and a solvent whereby the solvent does not interfere with the biological activity of the solute. Suitable solvents include, for example, ethanol, acetic acid, and the like.

[0213] The term "hydrate" means a solvate as defined above wherein the solvent is water.

[0214] Pharmaceutical Compositions: In any of the methods or uses described herein, the compounds of the present embodiments may be utilized as such or as a pharmaceutical composition in admixture with suitable carriers or excipients.

[0215] According to an aspect of some embodiments of the present invention there is provided a pharmaceutical composition comprising a compound described herein and a pharma- ceutically acceptable carrier.

[0216] As used herein, a "pharmaceutical composition" refers to a formulation of one or more active ingredients described herein with other chemical components, such as physiologically suitable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration of a compound to an organism.

[0217] The term "active ingredient" refers to a compound responsible for a biological action.

[0218] The phrases "physiologically acceptable carrier" and "pharmacologically acceptable carrier", used interchangeably hereinafter, refer to a carrier or diluent that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound. Such phrases include adjuvants.

[0219] In this application, the term "excipient" refers to an inert substance added to a pharmaceutical composition to further facilitate administration of an active ingredient. Non-limiting examples of excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils and polyethylene glycols.

[0220] Pharmaceutical compositions of some embodiments of the present invention may be manufactured by processes well known in the art, e.g., by conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or lyophilizing processes.

[0221] Thus, pharmaceutical compositions for use according to some embodiments of the present invention can be formulated in a conventional manner using one or more physiologically acceptable carriers, including excipients and adjuvants, which facilitate the processing of the active ingredient into a pharma- ceutically acceptable preparation. Appropriate formulations will vary depending on the selected route of administration.

[0222] Techniques for drug formulation and administration can be found in "Remington's Pharmaceutical Sciences," Mack Publishing Co., Easton, PA, latest edition, which is incorporated herein by reference.

[0223] Suitable routes of administration can include, for example, oral, rectal, transmucosal, particularly intranasal, intestinal or intramuscular, parenteral delivery including subcutaneous and intramedullary injection, as well as intrathecal, direct intraventricular, intracardiac (e.g., into the right or left ventricular cavity, into the common carotid artery), intravenous, intraperitoneal, intranasal or intraocular injection.

[0224] Alternatively, the pharmaceutical composition may be administered in a local rather than systemic manner, for example, by injection of the pharmaceutical composition into a tissue region of the patient.

[0225] The term "tissue" refers to a portion of an organism that is composed of cells designed to perform one or more functions. Examples include, but are not limited to, brain tissue, retina, skin tissue, liver tissue, pancreatic tissue, bone, cartilage, connective tissue, blood tissue, muscle tissue, heart tissue, brain tissue, vascular tissue, kidney tissue, lung tissue, gonadal tissue, and hematopoietic tissue.

[0226] According to some embodiments of the present invention, the compounds or pharmaceutical compositions described herein are administered topically.

[0227] For topical administration, a suitable carrier, and optionally other ingredients, can be selected in the composition, as described in detail herein, and the composition can thus be in the form of, for example, a cream, ointment, paste, gel, lotion and / or wash.

[0228] Ointments are typically semisolid preparations based on vegetable oils (e.g., shea butter and / or cocoa butter) or petrolatum or petrolatum derivatives. As with other carriers or vehicles, ointment bases should be inert, stable, nonirritating, and nonsensitizing.

[0229] Lotion is a preparation that is applied to the skin surface without friction.Lotion is typically a water-based or alcohol-based liquid or semi-liquid preparation, such as an oil-in-water emulsion.Lotion is generally preferred when treating large areas of the body (e.g., as is often desired in sunscreen compositions) because it is easier to apply a more fluid composition.

[0230] Creams are viscous liquids or semisolid emulsions, either oil-in-water or water-in-oil. Cream bases typically contain an oil phase, an emulsifier, and an aqueous phase. The oil phase, also called the "lipophilic" phase, optionally contains petrolatum and / or a fatty alcohol, such as cetyl or stearyl alcohol. The aqueous phase optionally contains a humectant. The emulsifier in a cream formulation is optionally a nonionic, anionic, cationic, or amphoteric surface active ingredient.

[0231] As used herein, the term "emulsion" refers to a composition that contains liquids in two or more distinct phases (e.g., a hydrophilic phase and a lipophilic phase), and that may optionally contain non-liquid substances (e.g., dispersed solids and / or gas bubbles).

[0232] As used herein and in the art, a "water-in-oil emulsion" is an emulsion characterized by an aqueous phase dispersed in a lipophilic phase.

[0233] As used herein and in the art, an "oil-in-water emulsion" is an emulsion characterized by a lipophilic phase dispersed in an aqueous phase.

[0234] Pastes are semi-solid dosage forms, and according to the nature of the base, they are divided into fatty pastes and pastes made from single-phase aqueous gels. The base of fatty pastes is generally petrolatum, hydrophilic petrolatum, etc. Pastes made from single-phase aqueous gels generally incorporate carboxymethylcellulose, etc. as a base.

[0235] Gel formulations are semi-solid, suspension-type systems. Single-phase gels contain organic macromolecules distributed substantially homogeneously throughout the carrier liquid, which is typically aqueous, but preferably also contains a non-aqueous solvent and, optionally, oil. Preferred organic macromolecules, i.e., gelling agents, include crosslinked acrylic acid polymers, such as carboxypolyalkylenes, available as those commercially available under the Carbopol® trademark, family of carbomer polymers. Other types of preferred polymers in this embodiment are hydrophilic polymers, such as polyethylene oxide, polyoxyethylene-polyoxypropylene copolymers, and polyvinyl alcohol; cellulosic polymers, such as hydroxypropylcellulose, hydroxyethylcellulose, hydroxypropylmethylcellulose, hydroxypropylmethylcellulose phthalate, and methylcellulose; gums, such as tragacanth and xanthan gum; sodium alginate; and gelatin. To prepare homogeneous gels, dispersing agents, such as alcohol or glycerin, may be added, and the gelling agent may be dispersed by grinding, mechanical mixing or stirring, or a combination thereof.

[0236] Compositions formulated for topical administration may optionally be included in a patch, swab, cotton ball and / or pad.

[0237] Such skin patches may contain all or some of the following components: a composition to be administered (e.g., as described herein), a liner to protect the patch during storage, which is optionally removed prior to use, an adhesive to hold the various components together and / or attach the patch to the skin, a backing to protect the patch from the external environment, and / or a membrane to control the release of drug into the skin.

[0238] According to some embodiments of the present invention, the compounds or pharmaceutical compositions described herein are administered to deliver the compounds to the central and / or peripheral nervous system.

[0239] Traditional approaches for drug delivery to the central nervous system (CNS) include neurosurgical strategies (e.g., intracerebral or intraventricular injections), molecular engineering aimed at engineering drugs to exploit one of the endogenous transport pathways of the BBB (e.g., fabrication of chimeric fusion proteins that include a combination of a transport peptide with affinity for an endothelial cell surface molecule and a drug that cannot cross the BBB by itself), pharmacological strategies designed to increase the lipid solubility of drugs (e.g., conjugation of water-soluble drugs to lipid or cholesterol carriers), and transient disruption of the integrity of the BBB by hyperosmotic disruption (by injecting a mannitol solution into the carotid artery or using biologically active drugs such as angiotensin peptides). However, each of these strategies has limitations, such as the inherent risks associated with invasive surgical procedures, size limitations imposed by the limitations inherent in endogenous transport systems, potentially undesirable biological side effects associated with systemic administration of chimeric molecules containing carrier motifs that may be active outside the CNS, and the risk of brain damage that may occur in regions of the brain where the BBB is disrupted, which result in suboptimal delivery methods.

[0240] Perineural injection or local administration via eye or ear drops are also contemplated for delivery of active ingredients to the central nervous system.

[0241] For injection, the active ingredient of the pharmaceutical composition can be formulated in an aqueous solution, preferably in a physiologically compatible buffer such as Hanks' solution, Ringer's solution or physiological salt buffer. For transmucosal administration, a penetrant appropriate to the barrier to be permeated is used in the formulation. Such penetrants are generally known in the art.

[0242] For oral administration, pharmaceutical compositions can be easily formulated by combining active compounds with pharma- ceutically acceptable carriers well known in the art. Such carriers allow pharmaceutical compositions to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, etc., for oral ingestion by patients. Pharmacological preparations for oral use can be made by using solid excipients, optionally grinding the mixture obtained, and processing the mixture into granules, after adding suitable excipients as necessary, to obtain tablets or dragee cores. Suitable excipients include, in particular, sugars including lactose, sucrose, mannitol or sorbitol, e.g., corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, cellulose preparations such as methylcellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and / or fillers such as physiologically acceptable polymers, e.g., polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.

[0243] Dragee cores are provided with suitable coatings.For this purpose, can be used concentrated sugar solutions, which can optionally contain gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures.Dyes or pigments can be added to tablets or dragee coatings for identification or to characterize different combinations of active compound doses.

[0244] Pharmaceutical compositions that can be used orally include push-fit capsules made of gelatin and soft sealed capsules made of gelatin and plasticizers such as glycerol or sorbitol. Push-fit capsules can contain active ingredients in a mixture with fillers such as lactose, binders such as starch, lubricants such as talc or magnesium stearate, and optionally stabilizers. In soft capsules, active ingredients can be dissolved or suspended in a suitable liquid such as fatty oils, liquid paraffin, or liquid polyethylene glycol. In addition, stabilizers can be added. All formulations for oral administration should be in a dosage suitable for the selected route of administration.

[0245] For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.

[0246] For administration by nasal inhalation, the active ingredient for use according to some embodiments of the present invention is conveniently delivered in the form of aerosol spray formulation from a pressurized pack or nebulizer with suitable propellant, for example, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane or carbon dioxide.In the case of pressurized aerosol, dosage unit can be determined by providing a valve to deliver a metered amount.Capsules and cartridges of, for example, gelatin, can be formulated to contain a powder mix of the compound and a suitable powder base, for example, lactose or starch, for use in dispenser.

[0247] The pharmaceutical compositions described herein may be formulated for parenteral administration, for example, by bolus injection or continuous infusion. Preparations for injection may be provided in unit dosage form, for example, in ampoules or multi-dose containers, optionally with the addition of preservatives. The compositions may be suspensions, solutions or emulsions in oily or aqueous media, and may contain formulatory agents such as suspending, stabilizing and / or dispersing agents.

[0248] Pharmaceutical compositions for parenteral administration include aqueous solutions of the active agent in water-soluble form.Furthermore, suspensions of active ingredient can be prepared as suitable oily or aqueous-based injection suspensions.Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil or synthetic fatty acid esters such as ethyl oleate, triglycerides or liposomes.Aqueous injection suspensions can contain agents that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol or dextran.Optionally, suspensions can also contain suitable stabilizers or agents that increase the solubility of active ingredient to allow the preparation of highly concentrated solutions.

[0249] Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, eg, sterile pyrogen-free water-based solution, before use.

[0250] Pharmaceutical compositions of some embodiments of the present invention may also be formulated in rectal compositions such as suppositories or retention enemas, using, for example, conventional suppository bases such as cocoa butter or other glycerides.

[0251] Pharmaceutical compositions suitable for use in connection with some embodiments of the present invention include compositions in which the active ingredient is contained in an amount effective to achieve the intended purpose. More specifically, a therapeutically effective amount refers to an amount of active ingredient (a compound described herein) effective to prevent, reduce or ameliorate symptoms associated with a condition described herein, or to prolong the survival of the subject being treated.

[0252] Determination of a therapeutically effective amount is well within the capabilities of those skilled in the art, especially in light of the detailed disclosure provided herein.

[0253] For any formulation used in the method of the present invention, therapeutically effective amount or dose can be estimated first from in vitro and cell culture assays.For example, dose can be formulated in animal models to achieve desired concentration or titer.Such information can be used to more accurately determine useful dose in humans.

[0254] The toxicity and therapeutic efficacy of the active ingredients described in this application can be determined by standard pharmaceutical procedures in vitro, in cell cultures, or in experimental animals. The data obtained from these in vitro and cell culture assays and animal studies can be used to determine a range of dosages for use in humans. The dosage can vary depending on the dosage form used and the route of administration utilized. The exact formulation, route of administration, and dosage can be chosen by the individual physician in view of the patient's condition (see, for example, Fingl et al. (1975) in "The Pharmacological Basis of Therapeutics", Ch. 1 p.1).

[0255] Dosage and intervals can be adjusted individually to provide tissue (e.g., plasma) levels of active ingredient sufficient to induce or inhibit a biological effect (minimal effective concentration, MEC). The MEC varies for each formulation but can be estimated based on in vitro data. Dosages required to achieve the MEC vary with individual characteristics and route of administration. Detection assays can be used to determine concentrations in the relevant tissues (e.g., plasma and / or brain).

[0256] In some of the embodiments of any of the present disclosure, the effective amount of the compound is less than 100 μM. In some embodiments, the effective amount is less than 10 μM. In some embodiments, the effective amount is less than 5 μM. In some embodiments, the effective amount is less than 1 μM. In some embodiments, the effective amount is less than 0.5 μM. In some embodiments, the effective amount is less than 0.1 μM.

[0257] In some of the embodiments of any of the present disclosure, the effective amount of the compound is within the range of 1 pM to 1 mM, or 1 pM to 100 μM, or 100 pM to 100 μM, or 100 pM to 10 μM, or 100 pM to 1 μM, or 100 pM to 500 nM, or 100 pM to 100 nM, including intermediate values ​​and subranges thereof.

[0258] In some of the embodiments of any of the present disclosure, the effective amount is at least 100% of the compound's IC50 for TRPV1 and / or Kv7.2 / 3. In some embodiments, the effective amount is at least 200% of the compound's IC50 for TRPV1 and / or Kv7.2 / 3. In some embodiments, the effective amount is at least 300% of the compound's IC50 for TRPV1 and / or Kv7.2 / 3. In some embodiments, the effective amount is at least 500% of the compound's IC50 for TRPV1 and / or Kv7.2 / 3. In some embodiments, the effective amount is at least 1000% of the compound's IC50 for TRPV1 and / or Kv7.2 / 3.

[0259] In some of the embodiments of any of the present applications, the effective amount of the compounds described herein is in the nM range (eg, 0.001 to 1,000 nM, or 0.001 nM to 100 nM).

[0260] In some of the embodiments of any of the present applications, the effective amount of the compounds described herein is at least 10% lower than the amount that results in hERG inhibition, or at least 20, 30, 40, 50, 60, 70, 80, 90, 100% lower, or even lower.

[0261] In some embodiments of the invention, the amount of a composition administered (e.g., a dosage or therapeutically effective amount of a compound described herein) required to achieve a therapeutic effect is at least 20% or at least 30% less than the amount of such a compound known to exhibit a similar therapeutic effect.

[0262] Depending on the severity and responsiveness of the condition to be treated, dosage may be of single or multiple administrations over the course of treatment lasting from several days to several weeks, or until a cure is achieved or the disease state is eliminated.

[0263] The amount of composition administered will, of course, be dependent on the subject being treated, the severity of the affliction, the manner of administration, the judgment of the prescribing physician, etc.

[0264] The compositions of some embodiments of the present invention may be provided in a pack or dispenser device, such as a kit approved by the FDA (United States Food and Drug Administration), if desired, and such a pack or device may contain one or more unit dosage forms containing the active ingredient. The pack may be, for example, a blister pack comprising metal or plastic foil. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser may also have a notice associated with the container in a form prescribed by a government agency regulating the manufacture, use, or sale of pharmaceuticals, reflecting that the composition form has been approved by that agency for administration to humans or animals. Such notice may be, for example, in the form of a label approved by the United States Food and Drug Administration for prescription drugs, or in the form of an approved product insert. Compositions comprising the formulations of the present invention, formulated in a compatible pharmaceutical carrier, may also be prepared, placed in a suitable container, and labeled for the treatment or diagnosis of an indicated condition, as detailed above.

[0265] The term "subject" as used herein includes mammals, preferably humans of any age or sex. According to certain embodiments, the term "subject" includes individuals at risk of developing a pathology.

[0266] The compounds or pharmaceutical compositions described herein may be provided alone or in combination with other active ingredients that are well known in the art to ameliorate a medical condition (e.g., neuropathic pain) and / or to activate potassium channels and / or inhibit TRPV1 activity as described herein.

[0267] In some embodiments, a compound or pharmaceutical composition described herein may be administered together with an activator of a potassium channel (e.g., Kv7.2 / 3) described herein, either as a co-formulation or as a separate formulation.

[0268] The pharmaceutical compositions may further comprise additional pharma- ceutical active or inactive agents (e.g., including, but not limited to, antibacterial agents, antioxidants, buffering agents, bulking agents, surfactants, anti-inflammatory agents, antiviral agents, chemotherapeutic agents, and antihistamines) and / or additional agents that can be used in the treatment of the conditions, diseases, or disorders described herein.

[0269] use: According to an aspect of some embodiments of the present invention there is provided a compound as described herein or a pharmaceutical composition as described herein for use in modulating the activity of a voltage-gated potassium channel.

[0270] According to an aspect of some embodiments of the present invention, there is provided a method of modulating activity of a voltage-gated potassium channel comprising contacting the potassium channel with a compound or pharmaceutical composition described herein, which can be accomplished in vitro, for example by contacting a cell, tissue or organ expressing the channel with the compound or composition, or in vivo, by administering a therapeutically effective amount of the compound or composition to a subject in need thereof.

[0271] In some embodiments, the potassium channel is Kv7.2 / 7.3 (alternatively referred to herein as Kv7.2 / 3).

[0272] In some embodiments, the modulation comprises opening potassium channels.

[0273] According to an aspect of some embodiments of the present invention there is provided a compound as described herein or a pharmaceutical composition as described herein for use in modulating the activity of a TRPV1 channel.

[0274] According to an aspect of some embodiments of the present invention, there is provided a method of modulating the activity of a TRPV1 channel, comprising contacting the TRPV1 channel with a compound or pharmaceutical composition as described herein, which can be performed in vitro, for example by contacting a cell, tissue or organ expressing the channel with the compound or composition, or in vivo by administering a therapeutically effective amount of the compound or composition to a subject in need thereof.

[0275] In some embodiments, modulating comprises inhibiting activity of the TRPV1 channel (blocking the channel).

[0276] According to an aspect of some embodiments of the present invention, there is provided a compound as described herein or a pharmaceutical composition as described herein for use in modulating the activity of both a voltage-gated potassium channel and a TRPV1 channel as described in any of the embodiments herein.

[0277] According to an aspect of some embodiments of the present invention, there is provided a method of modulating activity of voltage-gated potassium channels and TRPV1 channels, comprising contacting these channels with a compound or pharmaceutical composition described herein, which can be accomplished in vitro, for example by contacting a cell, tissue or organ expressing these channels with the compound or composition, or in vivo, by administering a therapeutically effective amount of the compound or composition to a subject in need thereof.

[0278] According to an aspect of some embodiments of the present invention there is provided a compound as described herein or a pharmaceutical composition as described herein for use in treating a condition associated with activity of a voltage-gated potassium channel and / or a TRPV1 channel.

[0279] According to an aspect of some embodiments of the present invention there is provided a compound as described herein or a pharmaceutical composition as described herein for use in treating a condition associated with activity of voltage-gated potassium channels and TRPV1 channels.

[0280] According to an aspect of some embodiments of the present invention, there is provided a method for treating a condition associated with activity of a voltage-gated potassium channel and / or a TRPV1 channel in a subject in need of such treatment, comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition as described herein in any of the embodiments and any combination thereof.

[0281] According to some of the embodiments described herein, the condition is one in which it would be beneficial to modulate the activity of one or, preferably both, voltage-gated potassium channels and TRPV1 channels, as described herein.

[0282] According to some of the embodiments described herein, the condition is one in which it would be beneficial to open voltage-gated potassium channels and inhibit (e.g., block) activity of TRPV1 channels, as described herein.

[0283] One example of a condition is neuropathic pain.

[0284] Any other pathology (condition, state, disease and / or disorder) associated with TRPV1 channel function and / or voltage-gated potassium channels as described herein is contemplated.

[0285] Examples of conditions that may be beneficially treated by the TRPV1 inhibitors (e.g., blockers) described herein (compounds having general formula I) include, but are not limited to, epilepsy, pain-related conditions such as neurogenic pain, neuropathic pain, allodynia, pain associated with inflammation, and pain associated with pancreatitis, bipolar disorder, mood disorders, psychotic disorders, schizophrenia, anxiety, tinnitus and motor neuron diseases, bladder overactivity, urinary incontinence, persistent visceral hypersensitivity, e.g., irritable bowel syndrome (IBD), chronic cough, and cancer (e.g., squamous cell carcinoma, prostate cancer, pancreatic cancer).

[0286] There are many disease states, conditions and disorders associated with defective potassium channel function. As with other potassium channel opening compounds, the compounds described herein are intended for use within the framework of the treatment of disease states, conditions, diseases and disorders associated with defective potassium channel function to treat, ameliorate, prevent, inhibit or limit the effects of disease states and conditions in animals, including humans.

[0287] Examples of conditions that can be beneficially treated by the potassium channel openers described herein include, but are not limited to, central or peripheral nervous system disorders, such as ischemic stroke, migraine, ataxia, Parkinson's disease, bipolar disorder, trigeminal neuralgia, convulsions, mood disorders, brain tumors, psychotic disorders, schizophrenia, pruritus, myokymia, neurogenic pain, neuropathic pain, seizures, epilepsy, tinnitus, hearing and vision loss, anxiety and motor neuron diseases.The compounds described herein can further be beneficially used as neuroprotectants (e.g., to prevent stroke, etc.).The compounds described herein are also useful for treating disease conditions such as gastroesophageal reflux disorder and gastrointestinal motility disorders.

[0288] The compounds disclosed herein can also be used as potential candidates for treating a variety of conditions in which inhibition of cortical and / or peripheral neuronal activity is beneficial, such as epilepsy, ischemic stroke, migraine, ataxia, myokymia, neurogenic pain, neuropathic pain, Parkinson's disease, bipolar disorder, trigeminal neuralgia, convulsions, mood disorders, psychotic disorders, schizophrenia, brain tumors, hearing and vision loss, anxiety, tinnitus, and motor neuron diseases.

[0289] According to some embodiment aspects of the invention, the compounds or compositions described herein are for use in inhibiting cortical and / or peripheral neuronal activity and / or for use in treating a condition in a subject in which inhibition of cortical and / or peripheral neuronal activity is beneficial, as described herein.

[0290] The compounds disclosed herein are particularly useful in the treatment of conditions associated with neuronal hyperexcitability.

[0291] In one aspect of some embodiments of the present invention, the compounds or compositions described herein are for treating a condition associated with hyperexcitation in a subject in need of such treatment.

[0292] In one aspect of some embodiments of the present invention, the compounds or compositions described herein are for use in the manufacture of a medicament for the treatment of a condition associated with hyperexcitation in a subject in need of such treatment.

[0293] In one aspect of some embodiments of the present invention, there is provided a method for treating a condition associated with hyperexcitation in a subject in need of such treatment, the treatment being carried out by administering to the subject an effective amount of a compound or composition described in the corresponding embodiment of the present application, in any combination.

[0294] Conditions associated with neuronal hyperexcitability include, but are not limited to, epilepsy, neurodegenerative diseases, neurodevelopmental disorders, stroke, retinal degenerative diseases, tinnitus, spinal cord injury, traumatic brain injury, hearing loss (due to excessive noise exposure or ototoxicity), neuropathic pain, attention deficit / hyperactivity disorder, autism, central pain syndromes, neurodegenerative diseases, multiple sclerosis, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Parkinson's disease, frontotemporal dementia, schizophrenia, Rasmussen's encephalitis, Huntington's disease, alcoholism or alcohol withdrawal, benzodiazepine withdrawal apace, neonatal convulsions, paroxysmal ataxia, muscle spasms, cerebral ischemia, cerebral palsy, asphyxia, anoxia, prolonged cardiac surgery, hypoglycemia, AIDS-related dementia, and anxiety disorders.

[0295] As used herein, "about" refers to ±10% or ±5%.

[0296] The terms "comprises," "comprising," "includes," "including," "having" and conjugations thereof mean "including but not limited to."

[0297] The term "consisting of" means "including and limited to."

[0298] The term "consisting essentially of" means that a composition, method, or structure may include additional ingredients, steps, and / or moieties, provided that the additional ingredients, steps, and / or moieties do not materially alter the basic and novel characteristics of the claimed composition, method, or structure.

[0299] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. For example, "a compound" or "at least one compound" includes a plurality of compounds, and may also include mixtures thereof.

[0300] Throughout this application, various embodiments of the invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity, and is not an inflexible limitation of the scope of the invention. Thus, the description of a range should be considered to specifically disclose all of the possible subranges, as well as each individual numerical value within that range. For example, the description of a range such as 1 to 6 is intended to specifically disclose not only the subranges 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., but also each individual numerical value within that range, e.g., 1, 2, 3, 4, 5, and 6. This applies regardless of the magnitude of the range.

[0301] Whenever a numerical range is given herein, it is intended to include any recited number (fractional or integer) within the range given. The phrases "range between" a first designated number and a second designated number and "range from" a first designated number to a second designated number are used interchangeably herein and are intended to include the first designated number and the second designated number, and all fractional and integer numbers therebetween.

[0302] As used herein, the term "method" means manner, means, techniques, and procedures for accomplishing a given task, including, but not limited to, those known to practitioners in the chemical, pharmacological, biological, biochemical, and medical fields or that can be readily developed by practitioners from known manners, means, techniques, and procedures.

[0303] The term "treating" as used herein in connection with an abnormal activity, disease or condition includes arresting, substantially inhibiting, slowing or reversing the progression of the condition, substantially ameliorating the clinical or cosmetic symptoms of the condition, or substantially preventing the worsening of the clinical or cosmetic symptoms of the condition.

[0304] The term "alkyl" as used herein means a saturated aliphatic hydrocarbon, including straight-chain and branched-chain groups. The alkyl group preferably has 1-20 carbon atoms. When a numerical range described herein is, for example, "1-20," this means that the group (in this case, the alkyl group) contains 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to 20 carbon atoms. A medium alkyl having 1-10 carbon atoms is more preferred. A low alkyl having 1-4 carbon atoms is most preferred. The alkyl group may be substituted or unsubstituted. When substituted, the substituents can be, for example, cycloalkyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, halo, carbonyl, thiocarbonyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, C-carboxy, O-carboxy, nitro, sulfonamido, trihalomethanesulfonamido, silyl, guanyl, guanidino, ureido, amino, or NR'R'', where R' and R'' are each independently hydrogen, alkyl, cycloalkyl, aryl, carbonyl, sulfonyl, trihalomethylsulfonyl, and complex 5- or 6-membered heteroalicyclic.

[0305] A "haloalkyl" group refers to an alkyl, as defined herein, substituted with one or more halo substituents, as defined herein. In some embodiments, the haloalkyl is an alkyl substituted with two or more or three or more halo substituents. In some embodiments, each of the halo substituents is fluoro. In some embodiments, the haloalkyl is -CF3 or -CF2H.

[0306] A "cycloalkyl" group refers to an all-carbon monocyclic or fused ring (i.e., rings which share adjacent pairs of carbon atoms) group in which one or more of the rings does not have a completely conjugated pi-electron system (alicyclic). Examples of cycloalkyl groups include, but are not limited to, cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, cyclohexadiene, cycloheptane, cycloheptatriene, and adamantane. Cycloalkyl groups can be substituted or unsubstituted. If substituted, the substituents can be, for example, alkyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, halo, carbonyl, thiocarbonyl, C-carboxy, O-carboxy, O-carbamyl, N-carbamyl, C-amido, N-amido, nitro, amino, and NR'R'' as defined herein.

[0307] An "alkenyl" group means an alkyl group composed of at least two carbon atoms and at least one carbon-carbon double bond.

[0308] An "alkynyl" group refers to an alkyl group composed of at least two carbon atoms and at least one carbon-carbon triple bond.

[0309] In some embodiments, when an alkyl substituent is indicated, it may be substituted with alkynyl or alkynyl as defined herein.

[0310] An "aryl" group refers to an all-carbon monocyclic or fused-ring polycyclic (i.e., rings which share adjacent pairs of carbon atoms) group having a completely conjugated pi-electron system. Examples of aryl groups include, but are not limited to, phenyl, naphthalenyl, and anthracenyl. Aryl groups can be substituted or unsubstituted. If substituted, the substituents can be, for example, halo, trihalomethyl, alkyl, hydroxy, alkoxy, aryloxy, thiohydroxy, thiocarbonyl, C-carboxy, O-carboxy, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, sulfinyl, sulfonyl, amino, and NR'R'' as defined herein.

[0311] A "heteroaryl" group refers to a monocyclic or fused ring (i.e., rings that share adjacent pairs of atoms) group having one or more atoms in the ring, such as, for example, nitrogen, oxygen, and sulfur, and further having a completely conjugated pi-electron system. Examples of heteroaryl groups include, but are not limited to, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrimidine, quinoline, isoquinoline, and purine. Heteroaryl groups can be substituted or unsubstituted. If substituted, the substituents can be, for example, alkyl, cycloalkyl, halo, trihalomethyl, hydroxy, alkoxy, aryloxy, thiohydroxy, thiocarbonyl, sulfonamido, C-carboxy, O-carboxy, sulfinyl, sulfonyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, amino, or NR'R'' as defined herein.

[0312] It should be noted that throughout this application, for example in Formulas II, III, IV and V, when pyridine is recited as the heteroaryl B ring, other nitrogen-containing heteroaryls are also contemplated.

[0313] A "heteroalicyclic" group refers to a monocyclic or fused ring group having one or more atoms in the ring, such as nitrogen, oxygen, and sulfur. The ring may also have one or more double bonds. However, the ring does not have a completely conjugated pi-electron system. Heteroalicyclic groups can be substituted or unsubstituted. If substituted, the substituents can be, for example, alkyl, cycloalkyl, aryl, heteroaryl, halo, trihalomethyl, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, carbonyl, thiocarbonyl, C-carboxy, O-carboxy, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, sulfinyl, sulfonyl, C-amido, N-amido, amino, and NR'R'' as defined herein.

[0314] As used herein, "cyclic group" refers to an alicyclic group (cycloalkyl), aryl, heteroaryl, or heteroalicyclic.

[0315] A "hydroxy" group refers to an --OH group.

[0316] An "azido" group refers to a -N=N group.

[0317] An "alkoxy" group refers to both an --O-alkyl group, and an --O-cycloalkyl group, as defined herein.

[0318] A "haloalkoxy" group refers to an O-alkyl group in which alkyl is haloalkyl as defined herein.

[0319] An "aryloxy" group refers to both an --O-aryl and an --O-heteroaryl group, as defined herein.

[0320] A "thiohydroxy" or "thiol" group refers to a --SH group.

[0321] A "thioalkoxy" group refers to both an --S-alkyl group, and an --S-cycloalkyl group, as defined herein.

[0322] A "thioaryloxy" group refers to both an --S-aryl group, and an --S-heteroaryl group, as defined herein.

[0323] A "carbonyl" group refers to a -C(=O)-R' group, where R' is hydrogen, alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon), or heteroalicyclic (bonded through a ring carbon) as defined herein.

[0324] An "aldehyde" group refers to a carbonyl group where R' is hydrogen.

[0325] A "thiocarbonyl" group means a -C(=S)-R' group, where R' is as defined herein.

[0326] The term "carboxylate" encompasses C-carboxylates and O-carboxylates.

[0327] A "C-carboxy" group refers to a -C(=O)-O-R' group, where R' is as defined herein.

[0328] An "O-carboxy" group refers to an R'C(=O)-O- group, where R' is as defined herein.

[0329] A "carboxylic acid" group refers to a C-carboxyl group where R' is hydrogen.

[0330] A "halo" group means fluorine, chlorine, bromine or iodine.

[0331] A "trihalomethyl" group refers to a -CX group, where X is a halo group as defined herein.

[0332] A "trihalomethanesulfonyl" group refers to a X3CS(=O)2- group, where X is a halo group as defined herein.

[0333] A "sulfinyl" group refers to a -S(=O)-R' group, where R' is as defined herein.

[0334] A "sulfonyl" group refers to a -S(=O)2-R' group, where R' is as defined herein.

[0335] The term "sulfonylamido" encompasses S-sulfonylamido and N-sulfonylamido.

[0336] An "S-sulfonamido" group refers to a -S(=O)2-NR'R'' group, where R' is as defined herein and R'' is as defined for R'.

[0337] An "N-sulfonamido" group refers to an R'S(=O)2-NR'' group, with R' and R'' as defined herein.

[0338] A "trihalomethanesulfonamido" group means a X3CS(=O)2NR'- group, with R' and X as defined herein.

[0339] The term "carbamate" includes O-carbamyl and N-carbamyl.

[0340] An "O-carbamyl" group refers to an --OC(.dbd.O)--NR'R'' group, with R' and R'' as defined herein.

[0341] An "N-carbamyl" group refers to an R'OC(=O)-NR''- group, with R' and R'' as defined herein.

[0342] The term "thiocarbamate" includes O-thiocarbamyl and N-thiocarbamyl.

[0343] An "O-thiocarbamyl" group means an --OC(.dbd.S)--NR'R'' group, with R' and R'' as defined herein.

[0344] An "N-thiocarbamyl" group means an R"OC(=S)NR'- group, with R' and R'' as defined herein.

[0345] An "amino" group refers to a --NR'R'' group, with R' and R'' as defined herein.

[0346] An "alkylamino" group refers to an amine group in which one of R' and R'' is alkyl (a monoalkylamine) or both R' and R'' are each independently alkyl (a dialkylamine).

[0347] The term "amide" encompasses C-amides and N-amides.

[0348] A "C-amido" group means a -C(=O)-NR'R'' group, with R' and R'' as defined herein.

[0349] An "N-amido" group refers to an R'C(=O)-NR'' group, with R' and R'' as defined herein.

[0350] A "quaternary ammonium" group is an --NHR'R'' group, where R' and R'' are independently alkyl, cycloalkyl, aryl, or heteroaryl. + means a group.

[0351] A "ureido" group refers to a -NR'C(=O)-NR''R''' group, where R' and R'' are as defined herein and R''' is as defined for R' or R''.

[0352] A "guanidino" group refers to a -R'NC(=N)-NR''R''' group, with R', R'', and R''' as defined herein.

[0353] A "guanyl" group refers to an R'R''NC(=N)- group, with R' and R'' as defined herein.

[0354] A "nitro" group refers to a -NO2 group.

[0355] A "cyano" group refers to a -C≡N group.

[0356] A "silyl" group refers to a -SiR'R''R''' group, where R', R'', and R'''' are as defined herein.

[0357] As used herein, the term "alkylene glycol" refers to an -O-[(CR'R'')zO]y-R''' terminal group or an -O-[(CR'R'')zO]y- linking group, where R', R'', and R''' are defined herein, z is an integer from 1 to 10, preferably 2 to 6, more preferably 2 or 3, and y is an integer equal to or greater than 1. Preferably, R' and R'' are both hydrogen. When z is 2 and y is 1, the group is ethylene glycol. When z is 3 and y is 1, the group is propylene glycol. When y is 2 to 4, the alkylene glycol is referred to herein as an oligo(alkylene glycol).

[0358] As used herein and in the art, "leaving group" refers to an unstable atom, group, or chemical moiety that is easily removed from an organic molecule during a chemical reaction, but removal is usually facilitated by the relative stability of the leaving atom, group, or moiety. Usually, any group that is the conjugate base of a strong acid can act as a leaving group. Representative examples of suitable leaving groups according to some of the present embodiments include, but are not limited to, trichloroacetimidate, acetate, tosylate, triflate, sulfonate, azide, halide (halo, preferably bromo or iodo), hydroxy, thiohydroxy, alkoxy, cyanate, thiocyanate, nitro, and cyano.

[0359] It will be understood that certain features of the invention that are, for clarity, described in the context of separate embodiments, may also be provided in a single embodiment in any combination of those features. Conversely, several features of the invention that are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination or with respect to other described embodiments as appropriate. Certain features described in the context of various embodiments should not be construed as essential to that embodiment, unless the particular embodiment is inoperable without that element.

[0360] As noted above, various embodiments and aspects of the present invention as described and claimed herein find experimental support in the following examples. EXAMPLES

[0361] Reference is now made to the following examples which, together with the above descriptions, illustrate the invention without limiting it.

[0362] Materials and Experimental Methods Solubility Test: To ensure accurate dose-response and reproducibility, solubility measurements were performed prior to each in vitro assay. Briefly, each compound was dissolved in 60 mM DMSO and a calibration curve was generated in the range of 1.5-200 μM using absorbance spectra and λmax determinations. The calibration curve served as the readout in all assays described below. The 60 mM stock solution was used to prepare a target concentration of 120 μM with 0.2% DMSO in the appropriate buffer for the in vitro related assays (i.e., HHBS buffer). The solubilized compounds were vortexed for 10 min, then centrifuged at 5000 RPM for 15 min, and the supernatant was collected for spectral analysis from 200-400 nm.

[0363] On-target assays: We used two kits that allow readouts for hKv7.2 / 3 and hTRPV1 in a high content screening (HCS) approach. Electrophysiology was used to validate the readouts of selected compounds that directly record modulation of ion channel currents, to study the inhibitory potential of our compounds on neuronal excitation, and to study the modulation of pH gating of TRPV1.

[0364] Kv7.2 / 7.3: Cell culture: Chinese hamster ovary (CHO) cell lines (B'SYS GmbH, Switzerland) constitutively expressing human Kv7.2 / 3 channels were cultured in F-12 nutrient mixture (Biological Industries) supplemented with 10% fetal bovine serum (Biological Industries) and 1% penicillin-streptomycin (Biological Industries) in a humidified 5% CO2 incubator at 37°C. Antibiotic selection was added (puromycin 5 μg / ml) to maintain the stability of human Kv7.2 / 7.3 expression.

[0365] High Content Screening (HCS) with FLIPR Potassium Assay Kit: To screen molecules against hKv7.2 / 7.3 ion channel targets, FLIPR Potassium Assay Kit (R8222 FLIPR Potassium Assay Explorer Kits, Molecular Devices) was used with CHO / hKv7.2 / 3 cell line. This assay exploits the permeability of thallium ions (Tl+) to potassium (K+) channels. In this assay, a Tl+ indicator dye is used to generate a bright fluorescent signal upon binding to Tl+ through the potassium channels. The intensity of the Tl+ signal is proportional to the number of potassium channels in the open state, thus providing a functional indication of potassium channel activity. To activate voltage-gated potassium channels, cells were stimulated with a mixture of K+ and Tl+ to depolarize the cell membrane. An increase in fluorescence in the assay represents the influx of Tl+ specifically through potassium channels and provides a functional readout of hKv7.2 / 3 activity using a fluorescent plate reader combined with the injector used to activate the channels.

[0366] Following an optimization process to achieve uniform and consistent screening conditions, the following protocol was established. 24 hours prior to performing the assay, cells were seeded at a density of 5000 cells per well in 384-well, black-walled, clear-bottom Greiner #781091 plates and cultured overnight in normal growth medium. On the day of the experiment, the medium was replaced with HBSS, HEPES, and test compound / vehicle (0.2% DMSO). Dye solution (prepared according to the manufacturer) was added to the plate and incubated at room temperature for 1.5 hours in the dark. For channel activation and data correction, the Tecan Spark reader plate injector was filled with Tl+ (1 mM Tl2SO4) or Tl+ and K+ (1 mM Tl2SO 4、5 mM K2SO4). Compound effects were compared to vehicle control and NH91 or retigabine positive controls. To calculate EC50 values, data were fitted to a sigmoidal regression using Prism GraphPad. The fit constrained the minimum to 1 and the maximum response to approximately 3.6 unless another maximum response was clearly identified.

[0367] Electrophysiology of Kv7.2 / 7.3: Extracellular (bath) solution: The solution consisted of the following (in mM): NaCl 140, KCl 4, CaCl2 1.8, MgCl2 1.2, glucose 11, HEPES 5.5. pH was adjusted to 7.3 with NaOH. Osmolarity was adjusted to 310 mOsm with sucrose.

[0368] Intracellular (pipette) solution: Pipettes were drawn from borosilicate glass (Warner Instrument Corp, USA) with resistances of 3–7 MΩ and filled with an internal solution consisting of the following (in mM): KCl 130, MgCl2 1, K ATP 5, EGTA 5, HEPES 10. pH was adjusted to 7.3 with KOH. Osmolarity was adjusted to 290 mOsm with sucrose.

[0369] A programmable, valve-linked pressurized perfusion system was used for local administration of compounds at a constant flow rate of 2-3 ml / min in the vicinity of the recorded cell. Series resistance was compensated for, and data were sampled at 5 kHz and low-pass filtered at 2.4 kHz using a MultiClamp 700B amplifier (Molecular Devices, USA) with pCLAMP11 software.

[0370] To assess the effect of test compounds on hKv7.2 / 3 currents at a membrane potential of -40 mV (threshold for action potential initiation), a -40 mV pulse train protocol was performed. The membrane potential was held at -90 mV and then clamped at -40 mV for 1.5 s, followed by a 0.75 s tail current acquisition with the membrane clamped at -60 mV and returned to a holding potential of -90 mV. A 30 s interval at a holding potential of -90 mV was maintained between sweeps.

[0371] Once a stable baseline current was achieved, test compounds were administered locally using a pressurized perfusion system until maximal and stable channel modulation at -40 mV was achieved, as confirmed by the recording of three resulting similar responses. Afterwards, cells were similarly perfused with the respective control bathing solution to assess the reversibility of the compound's effects.

[0372] To assess drug / control responses, the current obtained at a membrane potential of -40 mV upon injection of test compound was divided by the average of the currents at -40 mV recorded before administration of the test compound and following its washout.

[0373] TRPV1: Cell culture: Chinese hamster ovary (CHO) cell line cells (B'SYS GmbH, Switzerland) constitutively expressing human hTRPV1 channel were cultured in F-12 nutrient mixture (Biological Industries) supplemented with 10% fetal bovine serum (Biological Industries) and 1% penicillin-streptomycin (Biological Industries) in a humidified 5% CO2 incubator at 37°C. Antibiotic selection was added (G418, 500 μg / ml) to maintain the stability of hTRPV1 expression.

[0374] Species selection Rat, porcine, and human DNA constructs and DNA transfection: hTRPV1 cells were transfected with DNA constructs (Genscript). The coding regions of human (NM_080704.4), rat (NM_031982.1), and porcine (XM_013981216.2) TRPV1 were similarly inserted into the multiple cloning site of pcDNA3.1(+) between the HindIII and BamHI restriction sites. For transfection of CHO, 1x10 6Cells were suspended in a cuvette in a reaction mixture (Amaxa™ 4D-Nucleofector™-LONZA) containing 5 μg of DNA for each transfection. Cells were transformed (DT-133 program for CHO transfection with p-3 kit, Amaxa), allowed to recover for 10 min at room temperature in the hood, and allowed to settle by slowly transferring dropwise with a Lonza pipette into two wells of six wells containing warmed medium (no Ab). Following transfection, hTRPV1 cells were seeded into black, flat-bottom 384-well plates (Greiner #781091) the day before the experiment. Screening of molecules against CHO cells transiently transfected with rTRPV1 vs. hTRPV1 vs. pTRPV1 ion channels was performed with the Fluo-8 No Wash Calcium assay kit (ab112129, Abcam).

[0375] High content screening (HCS) using Fluo-8 No Wash Calcium flux assay kit: To screen compounds against the target of hTRPV1 ion channel, Fluo-8 No Wash Calcium assay kit (ab112129, Abcam) was used with CHO cells stably or transiently expressing TRPV1. Cells were preloaded with Fluo-8AM, which is membrane permeable. The AM group of Fluo-8AM is then cleaved by intracellular esterases, trapping Fluo8 inside the cells. Calcium influx through activated TRPV1 channels significantly increases the fluorescence of Fluo-8. Relative fluorescence signals are calculated after background subtraction, and the fluorescence at each time point is compared to the maximum measured after administration of ionomycin at the end.

[0376] Following an optimization process to achieve uniform and consistent screening conditions, the following protocol was established. 24 hours prior to performing the assay, cells were seeded into black-walled, clear-bottom 384-well plates at a density of 5000 cells per well and cultured overnight in normal growth medium. On the day of the experiment, the medium was replaced with HBSS, HEPES, and test compound / vehicle (0.2% DMSO). Dye solutions (prepared according to the manufacturer) were added to the wells and incubated for 1.5 hours at room temperature in the dark. Tecan Spark reader plate injectors were pretreated with capsaicin for channel activation and ionomycin for data correction. Compound effects were compared to vehicle controls and AMG9810 and NH91 positive controls. To calculate IC50 values, data were fitted to a sigmoidal regression using Prism GraphPad. Fitting constrained the minimum to 0 and the maximum response to approximately 1 unless another maximum response was clearly identified.

[0377] Electrophysiology of TRPV1: Extracellular (bath) solution: The solution consisted of the following (in mM): NaCl 140, KCl 4, CaCl2 1.8, MgCl2 1.2, glucose 11, HEPES 5.5. pH was adjusted to 7.3 with NaOH. Osmolarity was adjusted to 310 mOsm with sucrose.

[0378] Intracellular (pipette) solution: Pipettes were drawn from borosilicate glass (Warner Instrument Corp, USA) with resistances of 3–7 MΩ and filled with an internal solution consisting of the following (in mM): KCl 130, MgCl2 1, KATP 5, EGTA 5, HEPES 10. pH was adjusted to 7.3 with KOH. Osmolarity was adjusted to 290 mOsm with sucrose. A programmable, valve-linked pressurized perfusion system was used for local administration of compounds at a constant flow rate of 2–3 ml / min in the vicinity of the recorded cell. Series resistance was compensated for and data were sampled at 5 kHz and low-pass filtered at 2.4 kHz using a MultiClamp 700B amplifier (Molecular Devices, USA) with pCLAMP11 software.

[0379] The membrane potential was maintained at -60 mV. To measure the inhibitory effects of our test compounds on hTRPV1 activity, gating of hTRPV1 was achieved by fast, short-term administration of the hTRPV1 activator capsaicin (100 nM, 6 sec) with or without compound co-administration (over 3 min). Successive capsaicin injections were separated by 3 min intervals to allow for washing and cell recovery. All compounds were injected in the vicinity of the cells using a pressurized perfusion system.

[0380] To assess drug / control responses, drug current responses upon >3 min preincubation with test compound followed by capsaicin injection (coadministered with test compound) were divided by the mean control current obtained upon injection of capsaicin alone prior to compound incubation and following washout. To study pH activation of TRPV1, the following MES-based extracellular activation solutions at pH 5.5 were used:

[0381] Extracellular (bath) pH 5.5 activation solution: The solution consisted of the following (in mM): NaCl 140, KCl 4, CaCl2 1.8, MgCl2 1.2, glucose 11, MES 5.5. pH was adjusted to 5.5. Osmolarity was adjusted to 310 mOsm with sucrose.

[0382] Human NPCs: Cell Culture and Differentiation: Human PSC-derived neural progenitor cells (Stem Cell Catalog Nos. 70901 and 70902) were cultured and expanded on Matrigel-coated 6-well plates using neural progenitor medium supplemented with 1% Glutamax, 2% B27, and 20 ng / ml FGF2 in Neurobasal medium supplemented with 1% non-essential amino acids. hNPCs were then seeded onto 12 mm coverslips coated with PDL and Matrigel at a density of approximately 50,000 cells / well in 6-well plates and cultured in the presence of DMEM / F12, 10% KSR, 1% P / S, and A83-01 (2 μM) from days 0 to 5. From days 0 to 9, the medium additionally contained CHIR99021 (6 μM). From days 3 to 9, the medium contained RO4929097 (2 μM) and SU5402 (3 μM). From day 9 onwards, medium changes (Neurobasal medium containing NT-3, BDNF, NGF and GDNF) were performed every other day, with only 50% of the medium replaced after CO2 saturation. On day 12, cells were incubated with mitomycin C chemotherapy (2.5 μg / ml, 2 h, 37°C) to prevent glial cell proliferation.

[0383] Electrophysiology: Small, round neurons (20-40 μm diameter) were selected after day 31 to characterize neuronal properties by electrophysiological recordings. Cells were monitored for their polarized negative membrane potential and their ability to elicit consistent spike train responses to repeated positive current injections as a sign of achieving electrical maturation before testing the effects of test compounds on neuronal excitability. Membrane excitability was monitored using current clamp. Positive current steps of various amplitudes were injected (400 ms) to induce spike trains before and after compound administration.

[0384] Rat DRG: rDRG Isolation and Primary Cell Culture: In summary, rat DRGs from all levels of the spinal cord were carefully detached and collected in HBSS on ice, the epineural connective tissue surrounding the ganglion was removed, and the cells were dissociated using collagenase-II and trypsin dissociation solution. DRGs were further dissociated to obtain single cells through a glass Pasteur pipette. The dissociation solution was then replaced with DRG neuron culture medium containing 5% FBS and plated onto ECL-coated 12 mm coverslips.

[0385] Additional rDRG experiments were performed using Lonza primary neonatal rat (Sprague Dawley neonatal; P2,3) DRG neurons (R-DRG-505, Lonza), engrafted and cultured according to the manufacturer.

[0386] These cryopreserved DRG cells were prepared from freshly isolated and dissociated dorsal root ganglia and had a normal distribution of neurons and glial cells (Schwann cells).

[0387] Electrophysiology: Cells were used for electrophysiological recordings within 48 h of rDRG isolation. To amplify in vivo nociception and select small nociceptive neurons expressing both Kv7.2 / 3 and TRPV1 targets, small round neurons (20-40 μm diameter) were selected for electrophysiological recordings. Membrane excitability was monitored using current clamp. Positive current steps of various amplitudes were injected (400 ms) to induce spike trains before and after compound administration.

[0388] Off-target assays: Kv7.3 / 5, Kv7.4 assay: Cell Culture: Chinese hamster ovary (CHO) cell line cells were cultured in F-12 nutrient mixture (Biological Industries) supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin (Biological Industries) in a humidified 5% CO2 incubator at 37°C. To express the appropriate genes Kv7.3 / 5, Kv7.4 or TRPA1, 5 μg of plasmid was transfected using the DT-133 program from Amaxa with the p-3 kit.

[0389] FLIPR Potassium Assay Kit: The FLIPR Potassium Assay Kit (R8222 FLIPR Potassium Assay Explorer Kit, Molecular Devices) was used with the transfected CHO cells.

[0390] Following an optimization process to achieve uniform and consistent screening conditions, the following protocol was established. Two days after transfection and 24 hours before performing the assay, cells were seeded into black-walled, clear-bottom 384-well plates at a density of 5000 cells per well and cultured overnight in normal growth medium. On the day of the experiment, the medium was replaced with HBSS, HEPES, and test compound / vehicle (0.2% DMSO). Dye solution (prepared according to the manufacturer) was added to the wells and the plate was incubated at room temperature for 1.5 hours in the dark. For channel activation and data correction, the Tecan Spark reader plate injector was filled with Tl+ (1 mM Tl2SO4) or Tl+ and K+ (1 mM Tl2SO 4、 5 mM K2SO4). The effects of compounds were compared to vehicle control and NH91 or retigabine positive controls (known IC50 = 1.5-5 μM). To calculate IC50 values, data were fitted to a sigmoidal regression using Prism GraphPad.

[0391] TRPA1 Assay: Cell Culture: Chinese hamster ovary (CHO) cell line cells were cultured in F-12 nutrient mixture (Biological Industries) supplemented with 10% fetal bovine serum (Biological Industries) and 1% penicillin-streptomycin (Biological Industries) in a humidified 5% CO2 incubator at 37°C. For appropriate TRPA1 expression, 5 μg of plasmid was transfected using the DT-133 program from Amaxa with the p-3 kit.

[0392] High content screening (HCS) using Fluo-8 No Wash Calcium flux assay kit: To screen compounds against the target of hTRPA1 ion channel, Fluo-8 No Wash Calcium assay kit (ab112129, Abcam) was used with CHO cells transiently transfected with TRPA1. Cells were preloaded with Fluo-8AM, which is membrane permeable. The AM group of Fluo-8AM is then cleaved by intracellular esterases, trapping Fluo8 inside the cells. Calcium influx through activated TRPA1 channels significantly increases the fluorescence of Fluo-8. Relative fluorescence signals are calculated after background subtraction, and the fluorescence at each time point is compared to the maximum measured after administration of ionomycin at the end of the experiment.

[0393] Following an optimization process to achieve uniform and consistent screening conditions, the following protocol was established. Two days after transfection and 24 hours before performing the assay, cells were seeded into black-walled, clear-bottom 384-well plates at a density of 5000 cells per well and cultured overnight in normal growth medium. On the day of the experiment, the medium was replaced with HBSS, HEPES, and test compound / vehicle (0.2% DMSO). Dye solutions (prepared according to the manufacturer) were added to the wells and incubated for 1.5 hours at room temperature in the dark. Tecan Spark reader plate injectors were pretreated with AITC for channel activation and ionomycin for data correction. The effects of compounds were compared to vehicle control and A967079, a known inhibitor of TRPA1. To calculate IC50 values, the data were fitted to a sigmoidal regression using Prism GraphPad. The fit constrained the minimum to 0 and the maximum response to approximately 1 unless another maximum response was clearly identified.

[0394] hERG Assay: Cell culture: Chinese hamster ovary (CHO) cell line cells (B'SYS GmbH, Switzerland) constitutively expressing human hERG channels were cultured in F-12 nutrient mixture (Biological Industries) supplemented with 10% fetal bovine serum (Biological Industries) and 1% penicillin-streptomycin (Biological Industries) in a humidified 5% CO2 incubator at 37°C. Antibiotic selection (G418 200μg / ml and hygromycin 500) was applied to maintain the stability of hERG expression in the CHO cell line.

[0395] FLIPR Potassium Assay Kit: To screen compounds against hERG ion channel off-targets as described above in this application, a FLIPR Potassium Assay Kit (R8222 FLIPR Potassium Assay Explorer Kit, Molecular Devices) was used with the CHO / hERG cell line.

[0396] Following an optimization process to achieve uniform and consistent screening conditions, the following protocol was established. 24 hours prior to performing the assay, cells were seeded into black-walled, clear-bottom 384-well plates at a density of 7500 cells per well and cultured overnight in normal growth medium. On the day of the experiment, the medium was replaced with HBSS, HEPES, and test compound / vehicle (0.2% DMSO). Dye solution (prepared according to the manufacturer) was added to the wells and the plate was incubated at room temperature for 1.5 hours in the dark. For channel activation and data correction, the Tecan Spark reader plate injector was primed with Tl+ (1 mM Tl2SO4) or Tl+ and K+ (1 mM Tl2SO4, 5 mM K2SO4). The effects of compounds were compared to vehicle control and terfenadine positive control (known IC50=200 nM). To calculate IC50 values, data were fitted to a sigmoidal regression using Prism GraphPad.

[0397] HepG2 Assay: Cell Culture: HepG2 cell line cells were cultured in EMEM (ATCC 30-2033) medium supplemented with 10% fetal bovine serum (Biological Industries) at 37°C in a humidified 5% CO2 incubator.

[0398] ATPlite™ 1 Step Kit: The ATPlite™ 1 Step Kit (PerkinElmer) was used in a 384-well format to assess potential liver toxicity using HepG2 cells. It is an ATP detection system based on the luminescence of firefly luciferase to quantitatively assess cell proliferation and cytotoxicity. ATP is a marker of cell viability since it is present in all metabolically active cells and its concentration decreases rapidly upon cell necrosis or apoptosis.

[0399] Following an optimization process to achieve uniform and consistent screening conditions, the following protocol was established. 48 hours before performing the assay, cells were seeded into black-walled, clear-bottom 384-well plates at a density of 4000 cells per well and cultured overnight in normal growth medium. On the second day (48 hours before viability measurements), test compounds and positive controls were added to wells based on plate design. Compounds were dissolved in phenol-free growth medium to a DMSO concentration of 0.1% and their concentrations were identified using the solubility assay described in this application. After 48 hours, reconstituted lyophilized substrate solution was added to each well, the sealed 384-well microplate was mixed by rotating microplate at 1100 rpm for 2 minutes, and the plate was incubated at room temperature in the dark for 10 minutes. Immediately after, luminescence was measured with a Tecan plate reader. To estimate LD50 and LD25 values, the data was fitted to a sigmoidal regression using Prism GraphPad.

[0400] Drug Discovery Potential: SAR-driven optimization of metabolic stability and human liver microsome (HLM) levels: Metabolic stability refers to the susceptibility of a compound to biotransformation. In vitro half-life (t 1 / 2 Both the intrinsic clearance (CLint) and intrinsic clearance (CLint) are typically used to express metabolic stability. CLint represents the maximum activity of liver microsomal proteins / stem cells for a compound, without considering other physiological parameters such as hepatic blood flow and drug binding in the blood matrix. CLint is an important factor since the first pass metabolism of orally administered drugs is by the liver.

[0401] Liver microsomes, intracellular particles derived from the endoplasmic reticulum of hepatocytes, were used to evaluate test compounds. These particles are rich in drug metabolizing enzymes, including the cytochrome p-450 family. Liver microsomes are the recommended test system for a variety of in vitro drug metabolism and pharmacokinetic (DMPK) studies. CLint values ​​below 100 μl / min / mg are considered suitable for drug development.

[0402] LogD measurements: To accurately calculate the LLE and find compounds that exhibit improved physiochemical properties, the Log value of the partition coefficient between the lipid and aqueous phases of a chemical, LogD, was measured at pH 7.4 (equal to LogP).

[0403] LLE measurement: Ligand lipophilicity efficiency (LLE) is a parameter used to evaluate the properties, linking potency and lipophilicity of a investigated compound.

[0404] The following formula was used to track LLE values: LLE = -Log(IC50)-cLogP

[0405] Suitable LLE values ​​are typically greater than 3 or greater than 5.

[0406] Example 1 Structure-activity relationship (SAR) studies As explained above, one of the most potent compounds disclosed in WO 2019 / 073471 is called NH91.

[0407] NH91 has the following biological and physiochemical parameters: TRPV1 IC50: 8μM Kv7.2 / 7.3 EC50: 0.7μM CLogP: 5.3 LLE TRPV1: -0.35 LLE Kv7.2 / 7.3: 0.8 Solubility: 20μM

[0408] With the goal of improving the pharmaceutical potential of NH91, we focused on the pharmacophore sites of 4 as potential sites for modification: ring A, the bridging amine, ring B, and the side chain (meta to the amine bridge), as shown in Scheme 1 below.

[0409] Scheme 1 [ka]

[0410] A multi-layer preliminary library of compounds was synthesized and biological measurements and calculations (TRPV1 IC50, Kv7.2 / 7.3 EC50, LLE, kinetic solubility, etc.) were performed as described herein.

[0411] The preliminary library contained the following modifications: Modifications of Ring A: Replacement of the isopropyl group with other groups (tert-butyl, cyclopentane, pyridine, etc.) or replacement of one of the chloro substituents with, for example, another halo substituent, an electron-withdrawing substituent. Bridge modification: Replacement of the amine bridge with a sulfonamide or ether moiety (-O-). Modification of Ring B: Replacement of phenyl with pyridine. Side chain modifications: replacement of linear hydroxyalkyl with hydroxy-substituted cycloalkyl or heteroalicyclic, or branched hydroxyalkyl.

[0412] Data (not shown) obtained for compounds in this preliminary library showed the following: (i) Introducing bulky hydrophobic substituents such as tert-butyl and cycloalkyl into ring A improved the activity of both channels but did not improve the LLE. (ii) Introducing a bulky hydrophilic substituent into ring A improved the LLE but led to a decrease in activity. (iii) replacement of the amine bridge with a sulfonamide resulted in a substantial reduction in potassium channel opening. (iv) replacement of the amine bridge with an ether moiety improved parameters but reduced solubility; (v) Substitution of Ring B with pyridine resulted in acceptable parameters. (vi) Among the structures tested, 35 was modified in the side chain because previous studies have shown that both TRPV1 and Kv7.2 / 7.3 are sensitive to artificial modifications in this part of the molecule. The data obtained supported the LLE values ​​of these modifications as well as the desired effect on retaining activity and lower cLogP values.

[0413] Based on the data obtained for the preliminary library of compounds, additional libraries were designed, synthesized and tested containing one or two additional modifications at the pharmacological sites shown in Scheme 1. Additional designs included replacement of the phenyl at ring A and / or ring B with heteroaryls such as pyridine, imidazole or heteroalicyclics, replacement of the amide bridge with -S-, -S(=O)-, -P(OH)-, -CH(OH)-, or -C(=O)-, replacement of the cyclic A-NH-aniline moiety with a rigid heteroaromatic moiety such as indole, and further substitutions at the side chain sites.

[0414] Based on data obtained from additional libraries (not shown), three main chemotypes were shown to exhibit desirable properties in terms of dual activity towards TRPV1 and potassium channels, solubility, LLE and stability.

[0415] This chemotype included the following modifications to the crosslink: Chemotype 1: Ring A-NH-aniline moiety replaced with a rigid indole moiety; see Figure 1 for a representative structure. Chemotype 2: replacement of the amine bridge with -S- or -O-, see Figure 2 for representative structures. Chemotype 3: replacement of the amine bridge with -CH(OH)-, see Figure 3, A-B for representative structures. Chemotype 4: replacement of the amine bridge with -C(=O)-, see Figure 3 A-B for representative structures.

[0416] In addition, amide isosteres were also considered as replacements for the amide moiety in the side chain moiety. Exemplary such isosteres include oxetane structures (see, e.g., FIG. 1, compound 000526) and triazole structures (see, e.g., FIG. 1, compound 000527).

[0417] Additionally, compounds were explored that utilize heteroaryls (e.g., pyridines) as ring B. Exemplary such compounds of chemotype 1 include compounds 000661, 000662, 000663, and 000649 (see Figure 1).

[0418] The biological and physiochemical properties of representative compounds of each chemotype are shown in Table 1 below.

[0419] Note that chemotype 3 compounds (with dihydroxy structures) and the corresponding chemotype 4 compounds (with ketone bridges) are conceptually interesting because dihydroxy chemotype 3 can be oxidized to the corresponding ketone structure by the liver enzyme alcohol dehydrogenase (ADH) as shown in Figure 3B. Because the enzymatic reaction is reversible, the two structures may be generated as needed by the body. Under this scenario, dihydroxy chemotype 3 is expected to be primarily active against Kv7.2 / 7.3 (EC50=5.6 μM), whereas its associated in-situ oxidized ketone structure, chemotype 4, is expected to be primarily active against TRPV1 (IC50=8 μM).

[0420] [Table 1]

[0421] Example 2 Computer modeling Molecular docking and quantitative structure-activity relationship modeling (qSAR) represent the main computational tools used in computational chemistry. Molecular docking applies the 3D structure of a target protein to identify various molecules from their preferred active conformations within a selected virtual binding pocket (i.e., structure-based drug design). qSAR predicts the measured parameters of a compound by applying various physiochemical as well as 2D and 3D structural factors to a selected set of molecules (see Sharma, S., Recent trends in QSAR in modelling of drug-protein and protein-protein interactions. Comb. Chem. High Throughput Screen. 2020).

[0422] Molecular Docking: To perform docking experiments, a homology model of human TRPV1 was created. For this purpose, the following steps were performed. 1. Building multiple homology models: one using 5IRX (Prati, F. Stem Cell Translational medicine, 2017;6:369-381), the ligand-bound structure, and 3J5P (SA CAI, et al. Stem Cell Translational medicine, 2017;6:369-381), the apo structure ("closed conformation"). 2. Analyze the pockets in both and compare with the mutation data. 3. Identify lead compounds by docking for both homology models.

[0423] A homology model based on the published 3J5P model (SA CAI et al. 2017, supra) was generated (not shown), which further demonstrated a plausible binding mode and was consistent with the rat / human difference at 547 residues.

[0424] The homology model of human Kv7.2 / 7.3 was based on the open state of Kv7.1 (Peretz et al., Proceedings of the National Academy of Sciences Aug 2010, 107 (35) 15637-15642).

[0425] Figure 4A shows the docking of NH91 (000091, light yellow), the known TRPV1 inhibitor resiniferatoxin (RTX), and compound 228 (e.g., a compound described in WO 2004 / 035037) into the vanilloid pocket in the corresponding homology models. Figure 4B shows the docking of NH91 into the vanilloid pocket in the homology model of Kv7.2 / 7.3.

[0426] As can be seen, the hydrophobic gasket and hydrophilic channel are structural motifs common to both proteins, enabling the design of dual Kv7.2 / 7.3 and TRPV1 modulators.

[0427] Based on the three-dimensional structure of human TRPV1 and the favorable binding poses found for the test compounds (FIG. 6A), the following was deduced: = The newly designed compound has a high probability of binding to and interacting with the vanilloid pocket (i.e., capsaicin-binding pocket) of TRPV1; = Ring 'A' and Ring 'B' preferably interact with a hydrophobic surface area (i.e. a hydrophobic gasket); = Polar side chains are preferably located within the hydrophilic channels.

[0428] Based on the three-dimensional structure of human Kv7.2 / 7.3 and the favorable binding modes found for the test compounds (FIG. 4B), the following was deduced. = The hydrophobic A ring as well as the terminal hydroxyl (-OH) preferably interact with the hydrophilic channel and the hydrophobic gasket, respectively; = The aniline bridge results in a non-optimized rigid structure, indicating that modifications at this pharmacological site are a promising avenue for improved activity against Kv7.2 / 7.3.

[0429] Quantitative Structure-Activity Relationship Modeling (qSAR): To create such models, some of the hit compounds described in this application were superimposed on each other in their calculated 3D stable conformations to generate a pharmacophore representation, as shown in Figure 5A. Based on the pharmacophore representation, different learning models were generated that allow to correlate all IC50 / EC50 values ​​(generated for TRPV1 or Kv7.2 / 7.3, respectively) with their structural elements, as shown in Figure 5B for TRPV1.

[0430] Exemplary data of the TRPV1 qSAR modeling results are shown in Table 2.

[0431] [Table 2]

[0432] It is clearly shown that the representative TRPV1 qSAR model can highly correlate experimental IC50 values ​​with qSAR predictions.

[0433] Therefore, we constructed a computational infrastructure that consists of both homology models and docking capabilities as well as qSAR learning models.

[0434] Example 3 Efficacy Studies Effects on rat DRG neurons: The inhibitory potential of the newly designed compounds was studied by neuronal excitation in early neonatal DRG neurons (Sprague Dawley neonates; P2, 3; R-DRG-505, Lonza), a physiologically relevant experimental system. Cryopreserved DRG cells were prepared from freshly isolated and dissociated dorsal root ganglia and contained normal distribution of neurons and glia (Schwann cells).

[0435] The inhibitory potential was tested on both current-induced and capsaicin-induced neuronal activity, and the data obtained are shown in Figure 6, A–E.

[0436] As seen in Figure 6A, current stimulation identified the inhibitory potential of the test compounds as mediated through their effects on voltage-activated effectors such as Kv7.2 / 3 targets. As seen in Figure 6B, capsaicin-induced responses identified the contribution of capsaicin-gated TRPV1 targets to both membrane depolarization upon activation and subsequent activation of downstream voltage-gated Kv7.2 / 3 targets. Thus, by activating both targets, the additive effect of dual targeting of TRPV1 and Kv7.2 / 3 can only be compared to a single Kv7.2 / 3 target compound such as retigabine, which has a similar EC50 for Kv7.2 / 3 as 421-6.

[0437] More specifically, we first tested the ability of AMG9810 to inhibit both current- and capsaicin-induced neuronal activity. Administration of AMG9810 (50 nM) reduced capsaicin-induced firing by approximately 50%, whereas, predictably, current-induced responses were unchanged because AMG9810 does not target known voltage-dependent components of action potential firing.

[0438] Next, compound 273, which targets Kv only (Kv7.2 / 3 EC50=1.3 μM), was tested (2 μM) and showed a partial reduction in current-induced neuronal activity. Capsaicin-induced neuronal activity, which reads out a combination of both targets, showed a similar reduction in AP responses, supporting the effect of Kv activity as a general mechanism of reduction in action potential firing that is not dependent on the mechanism of action (current-induced or capsaicin-induced).

[0439] Next, we tested compound 421-6, which shows dual targeting of TRPV1 and Kv7.2 / 3 (Kv7.2 / 3 EC50=2.1 μM, TRPV1 IC50=1.8 μM). 421-6 administered at 2 μM, close to both its Kv7.2 / 3 EC50 and TRPV1 IC50, dramatically reduced AP responses to both current and capsaicin-induced activity, showing inhibition of 77.7 ± 5.6% (N=3) and 97.9 ± 2.1% (N=2), respectively. The marked inhibition of nerve voltage-dependent AP activity was higher than the 50% inhibition of Kv7.2 / 3 activity seen by measuring K influx into these channels expressed in heterologous systems. This may reflect changes arising from aspects that represent the distinct nature of these two readouts. One measures the gradual accumulation of ions passing through a gated ion channel target, while the other gives a physiologically relevant readout that is an all-or-none threshold event that is also controlled by the target. Possible different target interactions and compositions (e.g., stoichiometry and relative expression of different Kv subunits) in endogenous versus heterologous systems may explain these findings.

[0440] Given the apparent high potency of compound 421-6 in reducing neuronal firing, we tested its IC50 value for inhibiting firing responses in DRG neurons, since neural excitation is a physiologically predictable readout of pain amplification. Thus far, the activity of compound 421-6 has been tested at various nM concentrations.

[0441] The data obtained are shown in Figure 6A-E together with the comparative data obtained for retigabine (RET). Dilution of 421-6 significantly reduced its inhibition of current-induced AP responses, reaching its IC50 of approximately 500 nM (51.2 ± 8.4%, N = 3), while maintaining a significantly higher inhibition of capsaicin-induced AP responses, reflecting the effect of dual action on both targets (86.5 ± 8.4%, N = 4). Furthermore, 421-6 diluted to 100 nM inhibited current-induced AP activity by only 10.1 ± 3.4%, N = 2, while maintaining a high inhibitory capacity against capsaicin-induced AP responses (63.4 ± 17.8%, N = 3), highlighting the high potency and synergistic effect of this dual targeting compound.

[0442] Compound 421-6, a dual targeting compound, exhibits less than 100 nM inhibition of capsaicin-induced neuronal firing. This high inhibitory potency, significantly higher than the sum of the inhibition contributed by each target alone, indicates the synergistic effect of 421-6. This synergy exerts excellent specificity arising from (1) the high inhibitory potency for such compounds, and (2) the high activity inherent in pain sensory nerves, which occurs only when both targets are co-expressed and their signal pathways cross. Such synergy may occur when the second messenger of one target is a modulator of the other. For example, TRPV1 inhibition by the compounds of this embodiment inhibits Ca2+ influx through this channel, thereby relieving calcium-mediated inhibition of Kv7.2 / 3 channels, and may also synergistically activate M-currents with direct Kv7.2 / 3 activation by the same compounds.

[0443] 7A-C show data from these assays for representative compounds according to the present embodiments, designated compounds 552 and 541 (see FIG. 1) and 533 (see FIG. 3B).

[0444] Neuropathic model in rat DRG: NGF-treated DRGs showed bursts of tetanic action potentials in response to CAP administration, mimicking a neuropathic-like response. Prior art, see FIG. 8A. NGF promotes TRPV1 function using calcium-dependent fluorescence (F) versus maximum fluorescence (Fmax) as a function of time from a single HEK293 cell stably transfected with TrkA and transiently transfected with hTRPV1. A pulse of capsaicin (100 nM, administered as indicated above) induced a submaximal increase in [Ca]i. Exposure to NGF (100 ng / ml, see above) promoted the capsaicin-induced calcium increase. Arrows indicate responses used to calculate sensitization ratios.

[0445] As shown in Figure 8B-D, compound 421-6 attenuates capsaicin-induced tetanic action potential bursts to levels below the EC50, supporting the advantage of dual Kv / TRPV1 modulation. As 421-6 reaches a plateau concentration in the recording chamber, the second tetanic burst is modulated, as shown in Figure 8B. As concentrations are reduced by washout, the effect is maintained following a third dose of capsaicin, suggesting slow dissociation rate kinetics. This effect is reversible, as seen after the fourth capsaicin dose.

[0446] Effects on human NPCs: To study the inhibitory potential of exemplary lead compounds on neuronal excitation in an experimental system mimicking human pathophysiology, human neural progenitor cells (hNPCs) were differentiated into human sensory neurons and, following electrical maturation, their excitability was examined in the presence and absence of the test candidate.

[0447] Differentiated human sensory neurons underwent electrical maturation and exhibited a more negatively polarized resting membrane potential as they matured, as expected for mature neurons with an expanded population of K channels, shifting the resting membrane potential toward the potassium reversal potential.

[0448] The inhibitory potential of the novel designed compounds described herein was evaluated based on their ability to reduce spontaneous neuronal activity. As shown in Figure 9A, differentiated human sensory neurons exhibit reversibly reduced spontaneous activity following administration of chemotype 2 compound 415 (5μM). Compound 415 exhibits strong inhibitory potential in human sensory neurons to inhibit spontaneous firing, and further indicates the necessity to approach the target at much lower nM concentrations, which are predicted to be effective without possible side effects.

[0449] FIG. 9B shows data obtained from chemotype 2 compound 414 compared to compound 219, which has an aniline bridge. [ka]

[0450] These data demonstrate the potent inhibitory ability of the newly designed compounds described herein on the excitability of rat and human sensory neuronal cells.

[0451] Example 4 Further insight into the mechanism of operation Effects of racemic mixtures and separated enantiomers: To test whether different enantiomers of a molecule with a single chiral carbon show different activities, compound 533 was chosen as a representative example (* indicates a chiral center).

[0452] [ka]

[0453] The racemic mixture and the two enantiomers, designated 533p1 and 533p2, separated by SFC techniques, were tested in the potency study described in Example 3 above.

[0454] Figure 10A-C presents the inhibitory ability of test and reference compounds against current-induced and capsaicin-induced neuronal activity, with 533P1 showing primary activation of Kv7.2 / 3 and inhibition of TRPV1, whereas 533P2 shows stronger activation of Kv7.2 / 3 while activating TRPV1, indicating that stereospecific modification can be observed in 533 compounds. This is exemplified by current- and capsaicin-induced AP in rat DRG in Figure 10D, where 533P2 inhibits current-induced AP and enhances capsaicin-induced AP, whereas the racemic mixture combining the effects of the two enantiomers highly enhances capsaicin-induced AP inhibition while showing reduced current-induced AP inhibition.

[0455] Modification of dual crosstalk: Compound 627 (see FIG. 1 and Table 1) was selected to gain further insight into the mechanism of action.

[0456] CHO cells expressing hTRPV1 or co-expressing hTRPV1 and hKv7.2 / 3 were treated with compound 627 (see FIG. 1), AMG9810, a known TRPV1 inhibitor used as a positive control, or retigabine. The resulting data are shown in FIG. 11A-E.

[0457] Figure 11A-B present comparative plots depicting TRPV1 inhibition by compound 627 compared to retigabine. When hKv7.2 / 3 are co-expressed and activated by chemical depolarization, it can be seen that the potency of compound 627 increases significantly, shifting the dose-response curve to the left and becoming significantly more potent than the inhibitory potential of AMG9810.

[0458] Figures 11C-D provide plots showing inhibition of cells co-expressing hKv7.2 / 3, demonstrating that low concentrations of compound 627, even at picomolar concentrations, show significant hKv7.2 / 3 activation (Figure 11D).

[0459] These data demonstrate that compound 627 has a TRPV1 IC of 1.9 μM when administered to CHO cell lines expressing hKv7.2 / 3 or hTRPV1 alone and measured by fluorescent HCS. 50 and 0.5 μM Kv7.2 / 3 EC 50 We present here a method for determining whether Kv7.2 / 3 activity is reduced by approximately 30% at concentrations as low as 10 nM.

[0460] Activation of hKv7.2 / 3 activity by compound 627 was confirmed by electrophysiology by measuring whole-cell currents of hKv7.2 / 3 expressed in CHO cells and clamping the membrane potential to -40 mV (1.5 sec) (from a holding potential of -90 mV) (N=4), as seen in Figure 6E. Compound 627 at 100 nM demonstrated 24.4 ± 10.4% activation of Kv7.2 / 3 currents.

[0461] To illustrate the functional coupling of Kv7.2 / 3 and TRPV1, cells stably expressing Kv7.2 / 3 were transfected with human TRPV1 and their current ramp-up to -40 mV (Kv activation) and response to capsaicin (TRPV1 activation) were examined by electrophysiology. The data obtained are shown in Figure 12A-C.

[0462] The top panel of Figure 12A shows the effect of hKv7.2 / 3 co-expression in achieving potent 627 hTRPV1 inhibition. Although hTRPV1 expressing cells show partial TRPV1 inhibition by 1.3 μM compound 627, when hKv7.2 / 3 are co-expressed and activated (bottom panel), compound 627 achieves subnanomolar hTRPV1 inhibitory potency.

[0463] FIG. 12B presents a comparative dose-response curve showing the hTRPV1 inhibitory potency (shift to the left) gained in the presence of compound 627 when hKv7.2 / 3 are co-expressed and activated.

[0464] Figure 12C shows an exemplary current response to capsaicin in CHO cells co-expressing hTRPV1 and hKv7.2 / 3 but without hKv7.2 / 3 activation, demonstrating a lack of acquisition of hTRPV1 inhibitory capacity even at a concentration of 100 nM.

[0465] The data shown in Figure 12A-C show that the potentiation of TRPV1 inhibition increases by four orders of magnitude (from 2 μM to 0.26 nM) and requires Kv7.2 / 3. In addition, this potentiation of TRPV1 inhibition requires Kv7.2 / 3 activation, as even 100 nM was unable to inhibit TRPV1 without the prerequisite of current ramping to -40 mV (Kv activation).

[0466] FIG. 13A shows that in the presence of AMG9810, a known TRPV1 inhibitor, failure to acquire TRPV1 inhibitory capacity is observed in cells co-expressing hKv7.2 / 3 and hTRPV1, even when co-administered with retigabine, a known Kv7.2 / 3 opener.

[0467] FIG. 13B presents dose-response plots showing similar hTRPV1 inhibitory potency of AMG9810 and combination treatment of AMG9810 and retigabine, showing no synergistic effect between them. In contrast, compound 627 exerts different hTRPV1 inhibitory potency when hKv7.2 / 3 is co-expressed and activated, or when hTRPV1 is expressed alone. These data show that TRPV1 inhibition is uniquely affected by the same compound when Kv7.2 / 3 is co-expressed and activated.

[0468] 14 depicts an exemplary rat DRG membrane potential recording showing habituation of action potentials to capsaicin administration in the presence of compound 627. As shown in the figure, administration of compound 627 at a concentration of 1 nM completely blocked capsaicin-induced firing (top panel), and approximately 50% inhibition of action potential firing was observed when compound 627 was administered at a concentration of 0.1 nM (0.04 ng / ml).

[0469] While the present invention has been described in conjunction with specific embodiments thereof, many alternatives, modifications, and variations will be apparent to those skilled in the art. Accordingly, it is intended to embrace all such alternatives, modifications, and variations that fall within the spirit and broad scope of the appended claims.

[0470] It is the intention of the applicants that all publications, patents, and patent applications referenced herein be incorporated by reference as if each individual publication, patent, and patent application was specifically and individually indicated to be incorporated herein. In addition, citation or identification of any reference in this application should not be construed as an admission that such reference is available as prior art to the present invention. Nor should it necessarily be construed as limiting, to the extent that section headings are used. In addition, all priority documents of this application are incorporated by reference into this application.

Claims

1. The following formula I: 【Chemistry 1】 (In the formula, A and B are each independently selected from aryl and heteroaryl; W is -S-, -O-, -CR １ is selected from (OH)-, -C(=O), and NRx, where Rx together with one of the Ra substituents of Ring A forms a nitrogen-containing heterocycle; n is an integer from 1 to 5; m is an integer from 0 to 4; Ra and Rb are each independently a substituent selected from alkyl, cycloalkyl, halo, haloalkyl, haloalkoxy, alkoxy, ether, aryl, heteroaryl, heteroalicyclic, aryloxy, hydroxy, amine, alkylamine, thiohydroxy, thioalkoxy, thioaryloxy, cyano, carboxylate, amide, carbamate, sulfonyl, and sulfonamide, or alternatively, at least two of the Ra substituents taken together form an alicyclic or heterocyclic ring, at least one of the Ra substituents taken together with Rx, if present, forms a nitrogen-containing heterocyclic ring, and / or at least two of the Rb substituents taken together form an alicyclic or heterocyclic ring, when n is greater than 1, each Ra is the same or different substituent, and when m is greater than 1, each Rb is the same or different substituent; R １ is hydrogen or alkyl, if present; V is - (CR ２ R ３ ) k-U; k is 0, 1 or 2; R ２ and R ３ are each independently selected from hydrogen, alkyl, cycloalkyl, and aryl; U is an amide (-C(=O)-NR １０ -) or its isostere, The Z is represented by the following formula II: 【Chemistry 2】 (In the formula, u and q are each independently an integer from 0 to 4 (provided that u+q is at least 2); X is -O- and -NR ９ - or absent, Y is OR １１ , S.R. １１ , amine (NR １２ R １３ ) or amide (-NR １２ -C(=O)-R １４ ) and the like, R ５ , R ６ , R ７ and R ８ are each independently selected from hydrogen, halo, alkyl, haloalkyl, cycloalkyl, heteroalicyclic, aryl, alkylamino, alkoxy, haloalkoxy, hydroxy, ether, and aryloxy, or alternatively, R ５ , R ６ , R ７ , R ８ and R ９ two of which together form an alicyclic or heteroalicyclic ring; R ９ , R １０ , R １１ , R １２ , R １３ and R １４ are each independently selected from hydrogen, alkyl, cycloalkyl, and aryl, or alternatively, R ５ , R ６ , R ７ , R ８ , R ９ and R １１ Two of these, or R ５ , R ６ , R ７ , R ８ , R ９ , R １２ and R １３ Two of these together form an alicyclic or heteroalicyclic ring. A compound represented by the formula:

2. The following formula Ia: 【Chemistry 3】 (In the formula, W, V and Z are as defined in formula I; Ra1 to Ra5 ​​are each independently defined as Ra above, Rb1 to Rb4 each independently have the same definition as Rb above.) The compound according to claim 1 ,

3. 3. The compound of claim 1 or 2, wherein m is other than 0 and at least one of the Rb substituents is halo.

4. The following formula Ib: 【Chemistry 4】 (In the formula, W, V and Z are as defined in formula I; Rb1, Rb2 and Rb4 each independently have the same definition as Rb. Ra1 to Ra5 ​​are each independently defined as Ra above, The compound according to claim 1 ,

5. The compound according to any one of claims 1 to 4, wherein n is 3, 4 or 5.

6. The compound of claim 5, wherein at least two of the Ra substituents are selected from halo and alkoxy.

7. The compound according to any one of claims 1 to 6, wherein at least one of the Ra substituents is selected from alkyl, haloalkyl, cycloalkyl and aryl.

8. The compound according to any one of claims 1 to 6, wherein at least one of the Ra substituents is alkyl or cycloalkyl.

9. The compound according to claim 7 or 8, wherein at least one of the Ra substituents is ortho to the W.

10. n is 3; one of the Ra substituents is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamine, heteroaryl, or heteroalicyclic and is ortho to the W; the other two Ra substituents are each halo; m is 1, Rb is halo and is para to said W; The compound according to any one of claims 1 to 6.

11. The compound according to claim 2 or 3, wherein R a1 is an alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamine, heteroaryl, or heteroalicyclic.

12. 12. The compound of claim 2, 3, or 11, wherein Ra3 and Ra5 are each independently halo.

13. The compound of any one of claims 1 to 12, wherein X is absent.

14. R ５ , R ６ , , R ７ and R ８ At least one of R is independently selected from alkyl, hydroxy, alkoxy, haloalkyl, ether, and halo; and / or R ５ , R ６ , R ７ and R ８ The compound according to any one of claims 1 to 13, wherein at least two of the are taken together to form an alicyclic or heteroalicyclic ring.

15. 14. The compound of claim 13, wherein u is 1 or 2.

16. q is 1 and R ７ and R ８ At least one or each of R is alkyl; or R ７ and R ８ 16. The compound of claim 15, wherein at least one or each of is hydrogen.

17. R ５ , R ６ , R ７ and R ８ 16. The compound of claim 15, wherein at least one of is independently hydroxy, hydroxyalkyl, ether, heteroalicyclic (e.g., oxygen-containing), or cycloalkyl.

18. R ５ , R ６ , , R ７ and R ８ The compound according to any one of claims 1 to 13, wherein at least two of the following together form an alicyclic or heteroalicyclic ring, preferably an oxygen-containing 4-, 5- or 6-membered heteroalicyclic ring:

19. X is absent, u is 1 or 2, q is 1, and R ５ , R ６ , R ７ and R ８ taken together to form a cyclobutane or a furan.

20. Y is OR １１ and R １１ A compound according to any one of claims 1 to 19, wherein is hydrogen or alkyl, preferably alkyl substituted with at least one hydroxy, amido and / or carboxy, or heteroaryl, or heteroalicyclic.

21. Y is NR １２ R １３ and R １２ is R ５ , R ６ , R ７ and R ８ or Y is -NR １２ -C(=O)-R １４ and R １２ is R ５ , R ６ , R ７ and R ８ The compound according to any one of claims 1 to 19, which forms a nitrogen-containing heteroalicyclic ring with one or more of:

22. The compound according to any one of claims 1 to 21, wherein Rx together with one of the Ra substituents forms a nitrogen-containing heterocycle.

23. The compound according to claim 22, wherein Rx together with the Ra substituent ortho to W forms the nitrogen-containing heterocycle.

24. The following formula III: 【Chemistry 5】 (In the formula, V and Z are as defined above in formula I; D is N or C-Rb3; Ra2 to Ra5, Rb1 to Rb4, V and Z are as defined in formula I, Ia or Ib; Rc1 and Rc2 are each independently as defined above for Ra1 to Ra5. The compound according to claim 23, represented by:

25. 25. The compound of claim 24, wherein Rc1 and Rc2 are each independently selected from hydrogen, alkyl, haloalkyl, and halo.

26. The compound according to any one of claims 23 to 25, selected from the compounds shown in Figure 1.

27. The compound of any one of claims 1 to 21, wherein W is -S-.

28. Formula IV: 【Chemistry 6】 (In the formula, V and Z are as defined above in formula I; D is N or C-Rb3; Ra1 to Ra5, Rb1 to Rb4, V and Z are as defined in formula I, Ia or Ib. The compound according to claim 27, represented by:

29. 29. The compound of claim 28, wherein R a1 is selected from alkyl and cycloalkyl.

30. 29. The compound of claim 27 or 28, selected from the compounds shown in FIG.

2.

31. W is -CR １ The compound according to any one of claims 1 to 21, which is (OH)-.

32. The following formula V: 【Chemistry 7】 (In the formula, V and Z are as defined above in formula I; D is N or C-Rb3; Ra1 to Ra5, Rb1 to Rb4, V and Z are as defined in formula I, Ia or Ib. The compound according to claim 31 ,

33. 33. The compound of claim 32, wherein Ra1 is selected from alkyl and cycloalkyl.

34. The compound of any one of claims 1 to 21, wherein W is -C(=O)-.

35. The following formula VI: 【Chemistry 8】 (In the formula, V and Z are as defined above in formula I; D is N or C-Rb3; Ra1 to Ra5, Rb1 to Rb4, V and Z are as defined in formula I, Ia or Ib. The compound according to claim 34, represented by:

36. 36. The compound of claim 35, wherein R a1 is selected from alkyl and cycloalkyl.

37. The compound according to any one of claims 31 to 36, selected from the compounds shown in Figure 3A.

38. The compound according to any one of claims 1 to 37, characterized by at least one of the following: the LogD value, determined as described herein, is greater than 4, greater than 5, or within the range of 4 and 7; a ligand lipophilicity efficiency (LLE) greater than 3, or greater than 5, as determined according to the present application; HLM Clint is less than 100 μl / min / mg as determined according to the present application; The kinetic solubility is greater than 20 micromolar, or greater than 30 micromolar.

39. A pharmaceutical composition comprising a compound according to any one of claims 1 to 37 and a pharma- ceutically acceptable carrier.

40. A compound according to any one of claims 1 to 37 or a composition according to claim 39 for use in modulating the activity of a voltage-gated potassium channel.

41. A compound according to any one of claims 1 to 37 or a composition according to claim 39 for use in modulating the activity of TRPV1.

42. A compound according to any one of claims 1 to 37 or a composition according to claim 39 for use in modulating the activity of both voltage-gated potassium channels and TRPV1.

43. 43. The compound or composition of claim 42, wherein the modulation of the activity of the voltage-gated potassium channel comprises opening the channel and the modulation of the activity of the TRPV1 comprises inhibiting the activity of the TRPV1.

44. 44. A compound or composition according to claim 42 or 43, wherein the potassium channel is Kv7.2 / 7.

3.

45. A compound according to any one of claims 1 to 37 and 40 to 44 or a composition according to any one of claims 39 to 44 for use in treating a pathology associated with the activity of voltage-gated potassium channels and / or TRPV1 channels.

46. 46. ​​The compound or composition of claim 45, wherein the condition is neuropathic pain, pruritus or tinnitus.

Images