Modulators of potassium ion and TRPV1 channels and uses thereof

文档序号：1342965 发布日期：2020-07-17 浏览：26次中文

阅读说明：本技术 钾离子及trpv1通道的调节剂及其用途 (Modulators of potassium ion and TRPV1 channels and uses thereof ) 是由阿舍尔·佩雷斯伯纳德·阿塔利于 2018-10-09 设计创作，主要内容包括：本发明提供了多种用于调节一电压依赖性钾离子通道及/或TRPV1的一活性或功能的新颖化合物。所述多种化合物由说明书中描述及定义的式I表示。(The present invention provides novel compounds for modulating an activity or function of a voltage-dependent potassium channel and/or TRPV 1. The various compounds are represented by formula I as described and defined in the specification.)

1. A compound characterized by: represented by the following formula I:

wherein:

a and B are each independently selected from an aryl and a heteroaryl;

d is (CRdRe) u;

e is (CRfRg) v;

u and v are each independently 0 or 1;

n is an integer from 1 to 5;

m is an integer of 0 to 5;

re, Rd, Rf and Rg are each independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, halo, haloalkyl, haloalkoxy, alkoxy, aryl, heteroaryl, heteroalicyclic, aryloxy, hydroxy, amine, alkylamine, thiohydroxy, thioalkoxy, thioaryloxy, cyano, carboxylate, amide, carbamate, sulfonyl and sulfonamide;

ra and Rb are each independently a substituent selected from the group consisting of alkyl, cycloalkyl, halogen, haloalkyl, haloalkoxy, alkoxy, aryl, heteroaryl, heteroalicyclic, aryloxy, hydroxy, amine, alkylamine, thiohydroxy, thioalkoxy, thioaryloxy, cyano, carboxylate, amide, carbamate, sulfonyl and sulfonamide; or, alternatively, Ra substituents, Re, Rd and R₁At least two of which together form an alicyclic or heterocyclic ring; and/or at least two of the substituents Rb, Rf and Rg together form an alicyclic or heterocyclic ring, wherein 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;

R₁is hydrogen, alkyl, cycloalkyl or aryl; and

v is (CR)₂R₃)k-C(＝O)-NR₄-Z, and relative to said N-R₁In the meta position, wherein:

k is an integer from 0 to 2;

R₂and R₃Each independently selected from hydrogen, halogen, alkyl, cycloalkyl and aryl;

R₄is hydrogen, alkyl, cycloalkyl or aryl; and

z is represented by formula II:

wherein:

w and q are each independently an integer from 0 to 4, and provided that w + q is at least 2;

x is selected from O and NR₉Or absent;

y is selected from OR₁₀And SR₁₀；

R₅、R₆、R₇And R₈Each independently selected from the group consisting of hydrogen, halogen, alkyl, haloalkyl, cycloalkyl, heteroalicyclic, aryl, alkylamino, alkoxy, haloalkoxy, and aryloxy; or, alternatively, R₅、R₆、R₇、R₈And R₉Two of which together form an alicyclic or heteroalicyclic ring; and R₁₀Selected from hydrogen, alkyl, cycloalkyl and aryl; or, alternatively, R₅、R₆、R₇、R₈、R₉And R₁₀Two of which together form an alicyclic or heteroalicyclic ring, and with the proviso that:

at least one of Ra is selected from alkyl, haloalkyl, cycloalkyl and aryl; and/or

At least one of Rb is halogen; and/or

R₅、R₆、R₇And R₈At least one of which is an alkyl, cycloalkyl, heteroalicyclic or aryl group; and/or

R₅、R₆、R₇、R₈And R₉At least two of which together form an alicyclic or heteroalicyclic ring.

2. The compound of claim 1, wherein: a and B are each an aryl group.

3. The compound of claim 1, wherein: a and B are each phenyl.

4. A compound according to any one of claims 1 to 3, characterized in that: r₁Is hydrogen.

5. The compound according to any one of claims 1 to 4, characterized in that: u and v are each 0.

6. The compound of claim 1, wherein:

a and B are each phenyl;

R₁is hydrogen; and

u and v are each 0.

7. The compound according to any one of claims 1 to 6, characterized in that: m is not 0 and at least one of the Rb substituent(s) is halogen.

8. The compound of claim 7, wherein: m is 1.

9. The compound of claim 7 or 8, wherein: the halogen relative to the NR₁Is positioned at the contraposition.

10. A compound according to any one of claims 7 to 9, characterized in that: the halogen is fluorine.

11. A compound according to any one of claims 1 to 10, characterized in that: n is 3, 4 or 5.

12. The compound of claim 11, wherein: n is 3

13. A compound according to claim 11 or 12, characterized in that: at least two of the Ra substituents are selected from halogen and alkoxy.

14. The compound of claim 12 or 13, wherein: at least two of the plurality of Ra substituents are each halogen.

15. The compound of claim 14, wherein: the halogen is chlorine.

16. The compound according to any one of claims 1 to 15, characterized in that: at least one of the Ra substituent(s) is selected from the group consisting of alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl, and heteroalicyclic; alternatively, two Ra substituents together form a ring.

17. The compound according to any one of claims 1 to 15, characterized in that: at least one of the Ra substituent(s) is selected from the group consisting of alkyl, haloalkyl, cycloalkyl, and aryl.

18. The compound according to any one of claims 1 to 15, characterized in that: at least one of the Ra substituent(s) is alkyl or cycloalkyl.

19. A compound according to any one of claims 16 to 18, characterized in that: at least one of the Ra substituents relative to the NR₁In the ortho position.

20. According to any one of claims 1 to 15The compound is characterized in that: at least one of the Ra substituent(s) is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl or heteroalicyclic, and is related to the NR₁In the ortho position.

21. The compound according to any one of claims 1 to 15, characterized in that: n is 3, one of the Ra substituent(s) is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl or heterocycloaliphatic, and is in relation to the NR₁In the ortho position, and the other two Ra substituents are each halogen.

22. The compound according to any one of claims 1 to 15, characterized in that:

n is 3;

one of the Ra substituent(s) is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl or heteroalicyclic, and is relative to the NR₁In the ortho position;

the other two Ra substituents are each halogen;

m is 1; and

rb is halogen and relative to the NR₁Is positioned at the contraposition.

23. The compound according to any one of claims 1 to 22, characterized in that: k is 1.

24. The compound according to any one of claims 1 to 23, characterized in that: r₂And R₃Each is hydrogen.

25. The compound according to any one of claims 1 to 24, characterized in that: x is absent.

26. The compound according to any one of claims 1 to 25, characterized in that: x is O.

27. The compound according to any one of claims 1 to 26, wherein: r₅、R₆、R₇And R₈At least one of which is independently selected from alkyl, haloalkyl and halogen; and/or R₅、R₆R7 and R₈At least two of which together form an alicyclic ring.

28. The compound according to any one of claims 1 to 27, characterized in that: r₅、R₆、R₇And R₈At least two of which are independently selected from alkyl, haloalkyl and halogen.

29. The compound according to any one of claims 1 to 27, characterized in that: r₅、R₆、R₇And R₈At least two of which are each independently an alkyl group.

30. The compound according to any one of claims 1 to 29, characterized in that: q is 1, and R₇And R₈At least one or each of which is an alkyl group.

31. The compound of claim 30, wherein: w is 1 or 2.

32. The compound of claim 31, wherein: r₅And R₆Each is hydrogen.

33. The compound according to any one of claims 1 to 25, characterized in that: r₅、R₆、R₇And R₈At least two of which together form an alicyclic ring.

34. The compound according to any one of claims 27 to 29, wherein: at least one of the Ra substituent(s) is selected from the group consisting of alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl, and heteroalicyclic; alternatively, two Ra substituents together form a ring.

35. The compound according to any one of claims 1 to 34, characterized in that: y is OR₁₀And R is₁₀Is hydrogen.

36. The compound of claim 1, wherein: the compound is selected from:

and

37. the compound of claim 1, wherein: the compound is selected from:

and

38. the compound of claim 1, wherein: the compound is selected from:

and

39. a pharmaceutical composition, comprising:

a compound according to any one of claims 1 to 38; and

a pharmaceutically acceptable carrier.

40. The compound of any one of claims 1 to 38 or the composition of claim 39, wherein: for modulating an activity of a voltage-dependent potassium ion channel.

41. A compound or composition according to claim 40, wherein: the potassium ion channel is Kv7.2/7.3.

42. A compound or composition according to claim 40 or 41, wherein: the adjusting comprises: opening the channel.

43. The compound of any one of claims 1 to 38 or the composition of claim 39, wherein: for modulating an activity of TRPV 1.

44. A compound or composition according to claim 43, wherein: the adjusting comprises: inhibiting said activity of TRPV 1.

45. The compound of any one of claims 1 to 38 or the composition of claim 39, wherein: for modulating an activity of a voltage-dependent potassium channel and an activity of TRPV 1.

46. A compound or composition according to claim 31, characterised in that: modulating the activity of the voltage-dependent potassium ion channel comprises: opening the channel, and modulating the activity of the TRPV1 channel comprises: inhibiting an activity of the channel.

47. A compound or composition according to claim 45 or 46, wherein: the potassium ion channel is Kv7.2/7.3.

48. The compound of any one of claims 1 to 38 and 40 to 47, or the composition of any one of claims 39 to 47, wherein: for the treatment of a medical condition associated with an activity of a voltage-dependent potassium channel and/or a TRPV1 channel.

49. A compound of the composition of claim 48, wherein: the medical condition is neuropathic pain.

Technical field and background

The present invention relates to novel derivatives of diphenylamine, and more particularly, but not exclusively, to diphenylamine derivatives having dual activity as modulators of the potassium ion and TRPV1 channels, which are useful in the treatment of various pathologies associated with these channels, such as neuropathic pain.

Voltage-dependent potassium ion (Kv) channels conduct potassium ions (K) across cell membranes in response to changes in membrane voltage⁺) Thereby modulating cellular excitability by modulating (increasing or decreasing) the electrical activity of the cell.

A functional Kv channel exists as a multimeric structure formed by the association of four α and four β subunits. α subunits contain six transmembrane domains, a pore forming loop and a voltage sensor, and are symmetrically arranged around a central pore.the β or auxiliary subunits interact with α subunits and can modify the properties of the channel complex, including but not limited to changes in channel electrophysiological or biophysical properties, expression levels or expression patterns.

Functional Kv channels can exist as multimeric structures formed by the binding of identical or different Kv α and/or Kv β subunits.

Nine families of Kv channel α subunits have been identified, termed Kv1-Kv 9. thus, a great diversity in Kv channel function has emerged due to the diversity of subfamilies, the formation of identical and different subunits within a subfamily, and the additional impact of binding to the β subunit (m.j. christie, Clinical and experimental pharmacology and Physiology, 1995, 22(12), 944 951).

The Kv7 channel family consists of at least five members, including one or more of the following mammalian channels: kv7.1, kv7.2, kv7.3, kv7.4, kv7.5, and any mammalian or non-mammalian equivalents or variants thereof (including splice variants). Alternatively, the members of this family are referred to as KCNQ1, KCNQ2, KCNQ3, KCNQ4, and KCNQ5, respectively (Dalby-Brown et al, Current Topics in Medicinal Chemistry, 2006, 6, 999-.

The five members of this family are expressed differently. Expression of Kv7.1 is restricted to the heart, peripheral epithelium and smooth muscle, while expression of Kv7.2-Kv7.4 is restricted to the nervous system, including hippocampus, cortical and dorsal root ganglion neurons (for review, see, e.g., Delmas. P and Brown. D, Nature, 2005, 6, 850-.

Neuronal Kv7 channels have been shown to play a key role in controlling neuronal excitation. The Kv7 channel, particularly the Kv7.2/Kv7.3 heterodimer, is the basis for the M current (M-current), a non-activating potassium current found in many neuronal cell types. The current has a characteristic time and voltage dependence, stabilizing the membrane potential in response to a variety of excitatory stimuli. Thus, M currents are critical for controlling neuronal excitability (for review see, e.g., delmas. p and brown. d, Nature, 2005, 6, 850-.

Potassium ion channels have been associated with many physiological processes, including regulation of heart beat, arterial dilation, insulin release, neuronal excitability, and regulation of renal electrolyte transport. Thus, modulators of potassium ion channels are major drug candidates and the development of novel modulators as therapeutic agents is an ongoing research effort.

Therefore, given the key physiological role of Kv7 channels in the nervous system, and the relationship of these channels to a variety of diseases, it is highly desirable to develop modulators of Kv7 channels.

Potassium channel modulators are divided into channel openers and channel blockers. A potassium channel opener of particular interest is retigabine (retigabine, N- (2-amino-4- (4-fluorobenzylamino) -phenyl) carbamic acid ethyl ester). Retigabine is highly selective for KCNQ-type potassium ion channels composed of subunits kv7.2 and kv7.3, which was first described in EP0554543 in 1993. The use of retigabine for the treatment of neuropathic pain is disclosed, for example, in us patent No. 6,117,900 and EP 1223927. Compounds related to retigabine have also been proposed for use as mono-potassium ion channel modulators, see, e.g., U.S. patent No. 6,472,165.

However, retigabine has been reported to have multiple roles in neuronal cells. These include sodium and calcium channel blocking activity (Rundfeldt, C,1995, Nauyn-Schmiedberg's Arch Pharmacol,351 (suppl): R160), and effects on GABA (gamma-aminobutyric acid) synthesis and transmission in rat neurons (Kapetanovic, IM, 1995, Epilepsy Research (Epilepsy Research), 22, 167-.

Other KCNQ potassium channel modulators have been described, for example: in U.S. patent application No. 10/075,521, which teaches 2, 4-disubstituted pyrimidine-5-carboxamide derivatives as Kv7 modulators; U.S. patent application No. 10/160,582, which teaches cinnamamide derivatives as voltage-dependent potassium channel modulators; U.S. Pat. No. 5,565,483 and U.S. patent applications 10/312,123, 10/075,703 and 10/075,522, which teach 3-substituted oxindole derivatives as voltage-dependent potassium channel modulators; U.S. Pat. No. 5,384,330, which teaches 1,2, 4-triaminobenzene derivatives as potassium channel modulators; and U.S. patent No. 6,593,349, which teaches diarylamine derivatives as voltage-dependent potassium channel modulators. U.S. patent No. 6,291,442 teaches compounds containing two or three aromatic rings with a free carboxyl or a carboxyl group, which are linked by an ester bond to a lower alkyl ester linked to one of the rings, for modulating voltage-gated Shaker-type potassium ion channels.

WO2004/035037 and U.S. patent application publication No. 20050250833 teach N-phenyl anthranilic acid and 2-benzimidazolone derivatives as potassium channel openers, particularly voltage-dependent potassium channels, such as kv 7.2L, kv7.3 and kcv7.2/7.3 channels, and as modulators of neuronal activity.

WO 2009/037707 teaches other derivatives of N-phenyl anthranilic acid as potassium channel and/or TRPV1 modulators. An exemplary modulator disclosed in WO 2009/037707 is known as NH 29:

WO 2009/071947 and WO 2010/010380 teach derivatives of diphenylamine as potassium channel modulators. Exemplary modulators disclosed in WO 2009/037707 are known as NH34 and NH 43:

transient receptor potential vanilloid type 1 (TRPV 1) receptor is a ligand-gated non-selective cation channel activated by heat (typically above 43 ℃), low pH (<6), and products of endogenous lipid molecules such as arachidonic acid, N-acyl dopamine, and lipoxygenases (e.g., 12-and 15- (S) -HPETE), known as endo-vanilloids (endovanilloids). TRPV1 receptors are expressed throughout the brain except for the surrounding primary afferent neurons and the dorsal root ganglia. Recent evidence suggests that stimulation of TRPV1 receptors by endocannabinoids or capsaicin results in an analgesic effect, and that this effect is associated with glutamate augmentation and activation of OFF cells in the medullary cephalic intraabdominal (RVM).

TRPV1 was also found to be involved in the regulation of body temperature, anxiety and long-term depression in the hippocampus. TRPV1 channels are also located on sensory inputs innervating the bladder. Inhibition of TRPV1 has been shown to improve urinary incontinence symptoms.

TRPV1 modulators have been described, for example, in WO 2007/054480, which teaches the role of 2- (benzimidazol-1-yl) -acetamide derivatives in the treatment of TRPV1 related diseases. WO 2008/079683 teaches that the compounds are conjugated bicyclic ring systems of cyclohexyl and phenyl for inhibition of TRPV1 receptors. EP01939173 teaches O-substituted dibenzylurea or thiourea derivatives as TRPV1 receptor antagonists. WO 2008/076752 teaches benzimidazole compounds as potent TRPV1 modulators, and EP01908753 teaches TRPV1 modulators as heterocyclic subunit acetamide derivatives.

The potassium channel Kv7.2/3 and the cation non-selective channel TRPV1 are co-expressed in the major pain pathway of the peripheral nociceptive system (DRG sensory neurons), which transmits pain signals and has opposite functions. The TRPV1 channel triggers pain signals, whereas the Kv7.2/3 channel inhibits pain signals. Compounds that act as both kv7.2 openers and TRPV1 blockers can inhibit neuronal hyperexcitability, such as neuropathic pain.

Disclosure of Invention

According to an aspect of some embodiments of the present invention, there is provided a compound represented by formula I:

wherein:

a and B are each independently selected from an aryl and a heteroaryl;

d is (CRdRe) u;

e is (CRfRg) v;

u and v are each independently 0 or 1;

n is an integer from 1 to 5;

m is an integer of 0 to 5;

ra and Rb are each independently a substituent selected from the group consisting of alkyl, cycloalkyl, halogen, haloalkyl, haloalkoxy, alkoxy, aryl, heteroaryl, heteroalicyclic, aryloxy, hydroxy, amine, alkylamine, thiohydroxy, thioalkoxy, thioaryloxy, cyano, carboxylate, amide, carbamate, sulfonyl and sulfonamide; or, alternatively, Ra substituents, Re, Rd and R₁At least two of which together form an alicyclic ring (alicyclic ring) or a heterocyclic ring; and/or at least two of the substituents Rb, Rf and Rg together form an alicyclic or heterocyclic ring, wherein 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;

R₁is hydrogen, alkyl, cycloalkyl or aryl(ii) a And

v is (CR)₂R₃)k-C(＝O)-NR₄-Z, and relative to said N-R₁In the meta position, wherein:

k is an integer from 0 to 2;

R₂and R₃Each independently selected from hydrogen, halogen, alkyl, cycloalkyl and aryl;

R₄is hydrogen, alkyl, cycloalkyl or aryl; and

z is represented by formula II:

wherein:

w and q are each independently an integer from 0 to 4, and provided that w + q is at least 2;

x is selected from O and NR₉Or absent;

y is selected from OR₁₀And SR₁₀；

R₁₀selected from hydrogen, alkyl, cycloalkyl and aryl; or R₅、R₆、R₇、R₈、R₉And R₁₀Two of which together form an alicyclic or heteroalicyclic ring, and with the proviso that:

at least one of Ra is selected from alkyl, haloalkyl, cycloalkyl and aryl; and/or

At least one of Rb is halogen; and/or

R₅、R₆、R₇And R₈At least one of which is an alkyl, cycloalkyl, heteroalicyclic or aryl group; and/or

R₅、R₆、R₇、R₈And R₉At least two of which together form an alicyclic or heteroalicyclic ring.

According to some of any of the embodiments described herein, a is aryl.

According to some of any of the embodiments described herein, B is aryl.

According to some of any of the embodiments described herein, a and B are each an aryl group.

According to some of any of the embodiments described herein, a and B are each phenyl.

According to some of any of the embodiments described herein, R is₁Is hydrogen.

According to some of any of the embodiments described herein, u is 0.

According to some of any of the embodiments described herein, v is 0.

According to some of any of the embodiments described herein, each of u and v is 0.

According to some of any of the embodiments described herein, a and B are each phenyl; r₁Is hydrogen; u and v are both 0.

According to some of any of the embodiments described herein, m is not 0 and at least one of the Rb substituent(s) is halogen.

According to some of any of the embodiments described herein, m is 1.

According to some of any of the embodiments described herein, the halogen is relative to the NR₁Is positioned at the contraposition.

According to some of any of the embodiments described herein, the halogen is fluorine.

According to some of any of the embodiments described herein, n is 3, 4 or 5, preferably 3.

According to some of any of the embodiments described herein, at least two of the plurality of Ra substituents are selected from halogen and alkoxy.

According to some of any of the embodiments described herein, at least two of the plurality of Ra substituents are each halogen.

According to some of any of the embodiments described herein, the halogen is chlorine.

According to some of any of the embodiments described herein, at least one of the Ra substituent(s) is selected from the group consisting of alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl and heteroalicyclic; alternatively, two Ra substituents together form a ring.

According to some of any of the embodiments described herein, at least one of the Ra substituent(s) is selected from alkyl, haloalkyl, cycloalkyl and aryl.

According to some of any of the embodiments described herein, at least one of the Ra substituent(s) is alkyl or cycloalkyl.

According to some of any of the embodiments described herein, at least one of the plurality of Ra substituents is relative to the NR₁In the ortho position.

According to some of any of the embodiments described herein, at least one of the Ra substituent(s) is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl or heteroalicyclic, and is relative to the NR₁In the ortho position.

According to some of any of the embodiments described herein, n is 3, one of the Ra substituent(s) is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl or heteroalicyclic, and is relative to the NR₁In the ortho position, and each of the other two Ra substituentsIs a halogen.

According to some of any of the embodiments described herein, n is 3; one of the Ra substituent(s) is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl or heteroalicyclic, and is relative to the NR₁In the ortho position; the other two Ra substituents are each halogen; m is 1; and Rb is halogen and is relative to the NR₁Is positioned at the contraposition.

According to some of any of the embodiments described herein, k is 1.

According to some of any of the embodiments described herein, R is₂And R₃Each is hydrogen.

According to some of any of the embodiments described herein, X is absent.

According to some of any of the embodiments described herein, X is O.

According to some of any of the embodiments described herein, R is₅、R₆、R₇And R₈At least one of which is independently selected from alkyl, haloalkyl and halogen; and/or R₅、R₆、R₇And R₈At least two of which together form an alicyclic ring.

According to some of any of the embodiments described herein, R is₅、R₆、R₇And R₈At least two of which are independently selected from alkyl, haloalkyl and halogen.

According to some of any of the embodiments described herein, R is₅、R₆、R₇And R₈At least two of which are each independently an alkyl group.

According to some of any of the embodiments described herein, q is 1, and R is₇And R₈At least one or each of which is an alkyl group.

According to some of any of the embodiments described herein, w is 1 or 2.

According to the description hereinSome of any of the embodiments described above, R₅And R₆Each is hydrogen.

According to some of any of the embodiments described herein, R is₅、R₆、R₇And R₈At least two of which together form an alicyclic ring.

According to some of any of the embodiments described herein, Y is OR₁₀And R is₁₀Is hydrogen.

According to an aspect of some embodiments of the present invention there is provided a pharmaceutical composition comprising: a compound as described herein in any of the various embodiments and any combination thereof, and a pharmaceutically acceptable carrier.

According to an aspect of some embodiments of the present invention, there is provided a compound or composition as described herein in any of the various embodiments and any combination thereof, for modulating an activity of a voltage-dependent potassium ion channel.

According to some of any of the embodiments described herein, the potassium channel is kv 7.2/7.3.

According to some of any of the embodiments described herein, the adjusting comprises: opening the channel.

According to an aspect of some embodiments of the present invention there is provided a compound or composition as described herein in any of the various embodiments and any combination thereof, for modulating an activity of TRPV 1.

According to some of any of the embodiments described herein, said modulating comprises (blocking) inhibition of said activity of TRPV 1.

According to some of any of the embodiments described herein, modulating the activity of the voltage-dependent potassium ion channel comprises: opening the channel, and modulating the activity of the TRPV1 channel comprises: inhibiting an activity of the channel.

According to some of any of the embodiments described herein, the potassium channel is kv 7.2/7.3.

According to an aspect of some embodiments of the present invention, there is provided a compound or composition as described herein in any of the various embodiments and any combination thereof, for use in the treatment of a medical condition associated with an activity of a voltage-dependent potassium channel and/or a TRPV1 channel.

According to some of any of the embodiments described herein, the medical condition is neuropathic pain.

Unless defined otherwise, all technical and/or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the present invention, exemplary methods and/or materials are described below. In case of conflict, the patent specification and its definitions will control. In addition, the materials, methods, and examples are illustrative only and not intended to be necessarily limiting.

Drawings

Some embodiments of the invention are described herein, by way of example only, with reference to the accompanying drawings. With specific reference now to the drawings in detail, it is stressed that the particulars shown are by way of example and for purposes of illustrative discussion of the embodiments of the present invention. In this regard, the description taken with the drawings make it apparent to those skilled in the art how the embodiments of the invention may be embodied in practice.

In the drawings:

FIGS. 1A-1B present exemplary general synthetic routes for the preparation of the diphenylamine derivatives shown in Table A;

FIG. 2 presents an exemplary synthesis of NH 66;

FIG. 3 presents an exemplary synthesis of NH82 and NH 83;

FIG. 4 presents an exemplary synthesis of NH 91;

FIG. 5 presents an exemplary synthesis of NH 101;

FIG. 6 presents an exemplary synthesis of NH 110;

FIG. 7 presents an exemplary synthesis of NH 106;

FIGS. 8A to 8C present comparative graphs showing the effect of 5 μ M NH66 on the amplitude (FIG. 8A) and voltage dependence (FIG. 8B) of the recombined Kv7.2/3 current, as well as on the effect on the aforementioned diphenylamine compound NH43 (FIG. 8C);

FIG. 9 presents graphs showing the effect of 5 μ M NH66 on recombinant TRPV1 current activated by 0.1 μ M Capsaicin (CAP);

10A-10B present graphs showing the effect of 1 μ M NH43 (FIG. 10A) and 1 μ M NH66 (FIG. 10B) on induced spiking of rat DRG neurons;

11A-11D present comparative graphs showing the effect of NH82 on the amplitude (FIG. 11A) and voltage dependence (FIG. 11B) of recombined Kv7.2/3, and the effect of NH82 (FIG. 11C) and NH83 (FIG. 11D) on DRG-induced spiking;

FIGS. 12A to 12B present comparative graphs showing the effect of NH91 on the amplitude at-40 mV (FIG. 12A) and the voltage dependence (FIG. 12B) of the recombined Kv7.2/3 channel;

FIGS. 13A to 13C present comparative graphs showing the effect of NH101 on the amplitude at-40 mV (FIG. 13A) and the voltage dependence (FIGS. 13B and 13C) of the recombined Kv7.2/3 channel;

FIGS. 14A-14B present comparative plots showing the effect of NH110 on the amplitude at-40 mV (FIG. 14A) and voltage dependence (FIG. 14B) of the recombined Kv7.2/3 channel;

fig. 15A-15C present comparative graphs showing the inhibitory effect of NH91 on DRG-induced spiking (fig. 15A),and NH91(5 μ M) on inward voltage dependence of Ca in DRG neurons²⁺And Na⁺The current had no effect (fig. 15B to 15C);

fig. 16 presents a comparative graph showing NH110 suppression of DRG-induced spiking;

fig. 17A-17B present a comparison graph (fig. 17A) and a bar graph (fig. 17B) showing that NH91 produces a significantly more effective inhibition of capsaicin-induced DRG spikes compared to the known TRPV1 antagonist AMG 517;

figures 18A-18B present a comparison (figure 18A) and bar graph (figure 18B) showing that NH91 inhibits (50%) capsaicin-induced DRG spiking in the presence of kv7.2/3 channel blocker XE991(10 μ M);

FIG. 19 presents chemical structures and properties of exemplary compounds according to some embodiments of the invention; and

table a presents the chemical structures and molecular weights of the compounds tested in the screening assays as described in examples 1 and 2 below.

Detailed Description

In some embodiments, the present invention relates to novel derivatives of diphenylamine, and more particularly, but not exclusively, to diphenylamine derivatives having dual activity as modulators of both the potassium ion and TRPV1 channels, useful in the treatment of various pathologies associated with these channels, such as neuropathic pain.

Before explaining 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 set forth in the following description or illustrated by the examples. The invention is capable of other embodiments or of being practiced or carried out in various ways.

The present inventors have devised a "one-two punch" strategy that uses dual channel targeting to activate kv7.2 and inhibit TRPV1 through the same molecule to increase the potency of the drug (e.g., by additive/synergistic effects), allowing lower doses that would reduce the risk of off-target.

Embodiments of the present invention relate to newly designed compounds having a bis (aryl/heteroaryl) amide backbone (e.g., a diphenylamine backbone), e.g., represented by formula I as described herein in any of the various embodiments and any combination thereof, and to the use of these compounds in modulating an activity of a voltage-dependent potassium channel (e.g., kv7.2/7.3) and/or TRPV1, and treating medical conditions associated with an activity of these channels. Embodiments of the invention also relate to methods of making the disclosed compounds.

According to an aspect of some embodiments of the present invention, there is provided a compound represented collectively by formula I:

wherein:

a and B are each independently selected from an aryl and a heteroaryl;

d is (CRdRe) u;

e is (CRfRg) v;

u and v are each independently 0 or 1;

n is an integer from 1 to 5;

m is an integer of 0 to 5;

ra and Rb are each independently a substituent selected from the group consisting of alkyl, cycloalkyl, halogen, haloalkyl, haloalkoxy, alkoxy, aryl, heteroaryl, heteroalicyclic, aryloxy, hydroxy, amine, alkylamine, thiohydroxy, thioalkoxy, thioaryloxy, cyano, carboxylate, amide, carbamate, sulfonyl and sulfonamide; or, alternatively, Ra substituents, Re, Rd and R₁At least two of which together form an alicyclic or heterocyclic ring; and/or at least two of the Rb substituents, Rf and Rg together form an alicyclic or heterocyclic ring,wherein each Ra is the same or different substituent when n is greater than 1, and each Rb is the same or different substituent when m is greater than 1;

R₁is hydrogen, alkyl, cycloalkyl or aryl; and

v is (CR)₂R₃)k-C(＝O)-NR₄-Z, and relative to said N-R₁In the meta position, wherein:

k is an integer from 0 to 2;

R₂and R₃Each independently selected from hydrogen, halogen, alkyl, cycloalkyl and aryl;

R₄is hydrogen, alkyl, cycloalkyl or aryl; and

z is represented by formula II:

wherein:

w and q are each independently an integer from 0 to 4, and provided that w + q is at least 2;

x is selected from O and NR₉Or absent;

y is selected from OR₁₀And SR₁₀；

R₅、R₆、R₇And R₈Each independently selected from the group consisting of hydrogen, halogen, alkyl, haloalkyl, cycloalkyl, heteroalicyclic, aryl, alkylamino, alkoxy, and aryloxy; or, alternatively, R₄、R₅、R₆、R₇、R₈And R₉Two of which together form an alicyclic or heteroalicyclic ring (depending on R)₄、R₅、R₆、R₇、R₈And R₉The nature of the substituents forming a ring); and

R₁₀selected from hydrogen, alkyl, cycloalkyl and aryl; or, alternatively, R₅、R₆、R₇、R₈、R₉And R₁₀Two of which together form an alicyclic or heteroalicyclic ring. According to some implementations of the inventionIn one embodiment, a compound of the present embodiment is characterized by at least one, at least two, at least three, or all of the following:

at least one of the Ra substituents is selected from the group consisting of alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl, and heteroalicyclic; or, alternatively, two Ra substituents together form a ring; and/or

At least one of the Rb substituents is halogen; and/or

R₅、R₆、R₇And R₈Is alkyl, halogen, haloalkyl, cycloalkyl, heteroalicyclic or aryl; and/or

If present, R₅、R₆、R₇、R₈、R₉And R₁₀At least two of which together form an alicyclic or heteroalicyclic ring.

According to some embodiments of the invention, a compound of the present embodiments is characterized by at least one, at least two, at least three, or all of the following:

at least one of the Ra substituents relative to D (if present) or to NR₁(if no D is present) is in the ortho position and Ra in said ortho position is selected from the group consisting of alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl and heteroalicyclic; or, alternatively, the Ra substituent at the ortho position forms a ring with another Ra substituent, as described herein; and/or

At least one of the Rb substituents is halo and is located in the ortho position relative to the variable V; and/or

R₅、R₆、R₇And R₈Is alkyl, halogen, haloalkyl, cycloalkyl, heteroalicyclic or aryl; and/or

If present, R₅、R₆、R₇、R₈、R₉And R₁₀At least two of which together form an alicyclic or heteroalicyclic ring.

According to some embodiments of the invention, a compound of the present embodiments is characterized by at least one, at least two, at least three, or all of the following:

at least one of the Ra substituents relative to D (if present) or to NR₁(if no D is present) is in the ortho position and Ra in said ortho position is selected from alkyl, haloalkyl, cycloalkyl and aryl; and/or

At least one of the Rb substituents is halogen, preferably fluorine, and is in the ortho position relative to the variable V position; and/or

R₅、R₆、R₇And R₈Is alkyl, halogen, haloalkyl, cycloalkyl, heteroalicyclic or aryl; and/or

R₅、R₆、R₇And R₈At least two of which together form an alicyclic or heteroalicyclic ring.

According to some of any of the embodiments described herein, a is aryl.

According to some of any of the embodiments described herein, B is aryl.

According to some of any of the embodiments described herein, a is aryl and B is aryl or heteroaryl (e.g. pyridine).

According to some of any of the embodiments described herein, a and B are each an aryl group.

According to some of any of the embodiments described herein for a and/or B, the aryl is phenyl.

According to some of any of the embodiments described herein for a and/or B, the heteroaryl ring is pyridine.

When one or both of A and B is a heteroaryl group, one or more heteroatoms of the heteroaryl group may be relative to the D-NR linking the two rings₁The E group is in any position.

According to some of any of the embodiments described herein, a and B are each phenyl.

According to some of any of the embodiments described herein, D is (CRdRe) u, and Rd and Re are each hydrogen. Optionally, one or both of Rd and Re is a mono-alkyl, preferably a mono-unsubstituted alkyl, preferably a mono-lower alkyl, as defined herein, preferably a mono-lower unsubstituted alkyl. In some of these embodiments, u is 1. In some embodiments, u is 0 and D is absent.

According to some of any of the embodiments described herein, E is (CRfRg) v, and Rf and Rg are each hydrogen. Optionally, one or both of Rf and Rg are a mono-alkyl, preferably a mono-unsubstituted alkyl, preferably a lower alkyl, as defined herein, preferably a lower unsubstituted alkyl. In some of these embodiments v is 1. In some embodiments, v is 0 and E is absent.

According to some of any of the embodiments described herein, each of u and v is 0 and neither D nor E is present.

According to some of any of the embodiments described herein, a and B are each phenyl, and D and E are both absent.

The compounds according to these embodiments are diphenylamine derivatives and may be collectively represented by formula Ia:

wherein Ra, Rb, n, m, R₁And X is as described for formula I and in any of the various embodiments and any combination thereof.

In some embodiments of any of the embodiments described herein (e.g., for formulas I and Ia), R₁Is hydrogen or an alkyl group, preferably a mono-unsubstituted alkyl group, preferably a mono-lower alkyl group, as defined herein, preferably a mono-lower unsubstituted alkyl group.

In some embodiments of any of the embodiments described herein (e.g., for formulae I and Ia), R is₁Is hydrogen.

According to some embodiments of any of the embodiments described herein, there is at least one substituent on the B ring such that m is not 0 (e.g. is 1,2, 3 or 4).

According to some of these embodiments, in formula I or Ia, at least one of the plurality of Rb substituents is relative to-E-NR₁The radical-D is in para position, or if E is absent, relative to-NR₁-in the para position; or in an ortho position relative to V.

According to some embodiments of any of the embodiments described herein, m is not 0 and at least one of the Rb substituents is halogen.

According to some embodiments of any of the embodiments described herein, in formula I or Ia, m is not 0, and at least one of the Rb substituent(s) is halogen, and the halogen substituent is relative to-E-NR₁The radical-D is in para position, or if E is absent, relative to-NR₁-in the para position; or in an ortho position relative to V.

According to some of any of the embodiments described herein, m is 1 such that there is one substituent on the B ring in addition to V.

According to some of these embodiments, the Rb substituent is halogen.

According to some of these embodiments, in formula I or Ia, the Rb substituent is relative to-E-NR₁The radical-D is in para position, or if E is absent, relative to-NR₁-in the para position; or in an ortho position relative to V.

According to some embodiments of any of the embodiments described herein, in formula I or Ia, m is 1 and the Rb substituent is halogen, and with respect to-E-NR₁The radical-D is in para position, or if E is absent, relative to-NR₁-in the para position; or in an ortho position relative to V.

According to some embodiments of any of the embodiments described herein, when m is 1 and Rb is halogen, the halogen is fluorine.

According to some embodiments of any of the embodiments described herein, in formula I or Ia, m is 1, Rb is fluoro, and relative to-E-NR₁The radical-D is in para position, or if E is absent, relative to-NR₁-in the para position; or in an ortho position relative to V.

According to some embodiments of any of the embodiments described herein, m is not 0, such that one or more Rb substituents are present on the B ring, and the one or more Rb substituents may be the same or different from each other. In some of these embodiments, in formula I or Ia, one of the plurality of Rb substituents is relative to-E-NR₁The radical-D is in para position, or if E is absent, relative to-NR₁-in the para position; or in an ortho position relative to V.

In some embodiments, one or more Rb substituents may be, for example, selected from halogen, alkyl, haloalkyl, cycloalkyl, heteroalicyclic, aryl, heteroaryl, cyano, alkylamine, and the like, or two Rb substituents may together form a ring, as defined herein.

According to some embodiments of the invention, m is 1, and the Rb substituent may be halogen, alkyl, haloalkyl, cycloalkyl, heteroalicyclic, aryl, heteroaryl, cyano, alkylamine. According to some of these embodiments, in formula I or Ia, the Rb substituent is relative to-E-NR₁The radical-D is in para position, or if E is absent, relative to-NR₁-in the para position; or in an ortho position relative to V.

According to some embodiments of any of the embodiments described herein, at least one Ra substituent is present on the a ring in formula I such that n is 1,2, 3, 4 or 5.

According to some embodiments of any of the embodiments described herein, at least one of the plurality of Ra substituents is relative to-D-NR₁The E group being in the ortho position, or, in the absence of D, relative to-NR₁The radicals are in the ortho position.

According to some embodiments of any of the embodiments described herein, there are at least two Ra substituents on the a ring such that n is 2, 3, 4 or 5. According to some of these embodiments, at least one of the plurality of Ra substituents is relative to-D-NR₁The E group being in the ortho position, or, in the absence of D, relative to-NR₁The radicals are in the ortho position.

According to some embodiments of any of the embodiments described herein, there are at least three Ra substituents on the a ring such that n is 3, 4 or 5. According to some of these embodiments, at least one of the plurality of Ra substituents is relative to-D-NR₁The E group being in the ortho position, or, in the absence of D, relative to-NR₁The radicals are in the ortho position.

According to some embodiments of any of the embodiments described herein, there are three Ra substituents on the a ring such that n is 3. According to some of these embodiments, at least one of the plurality of Ra substituents is relative to-D-NR₁The E group being in the ortho position, or, in the absence of D, relative to-NR₁The radicals are in the ortho position.

According to some embodiments of any of the embodiments described herein, the Ra substituent may be a halogen, an alkoxy, a haloalkyl, an alkyl, a cycloalkyl, an amine (preferably: an alkyl amine), a heteroalicyclic, an aryl, and a heteroaryl; and, when n is 2, 3, 4, 5, any combination of the foregoing, optionally, two Ra substituents form a ring. According to some of these embodiments, at least one of the plurality of Ra substituents is relative to-D-NR₁The E group being in the ortho position, or, in the absence of D, relative to-NR₁The radicals are in the ortho position.

According to some embodiments of any of the embodiments described herein, when n is 2, 3, 4 or 5, and preferably 3, two or more Ra substituents selected from halogen and alkoxy, i.e. the Ra substituents may comprise two halogen substituents, two alkoxy substituents, or one halogen substituent and one alkoxy substituent. In some of any of the embodiments described herein, when an Ra substituent is alkoxy, the alkoxy is monohaloalkoxy as defined herein.

According to some embodiments of any of the embodiments described herein, n is 2, 3, 4 or 5, preferably 3, and at least two Ra substituents are each halogen.

According to some embodiments of any of the embodiments described herein, n is 2, 3, 4 or 5, preferably 3, and at least two Ra substituents are each chloro. Alternatively, two or more halogen substituents are fluoro.

According to some embodiments of any of the embodiments described herein, at least one of the Ra substituent(s) is selected from the group consisting of alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl, and heteroalicyclic; alternatively, two Ra substituents together form a ring.

According to some embodiments of any of the embodiments described herein, at least one of the Ra substituent(s) is selected from alkyl, haloalkyl, cycloalkyl and aryl.

According to some embodiments of any of the embodiments described herein, at least one of the Ra substituent(s) is alkyl or cycloalkyl.

According to some embodiments of any of the embodiments described herein, at least one of the Ra substituent(s) is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl or heteroalicyclic, and is of the-D-NR series₁The E group being in the ortho position or, if D is absent, relative to-NR₁The radicals are in the ortho position.

According to some embodiments of any of the embodiments described herein, n is 3, one of the Ra substituent(s) is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl or heteroalicyclic, and is of the-D-NR' s₁The E group being in the ortho position or, if D is absent, relative to-NR₁The radicals are in the ortho position and the two other Ra substituents are each halogen (e.g.chlorine).

According to some embodiments of any of the embodiments described herein, when n is 2, 3, 4 or 5, at least one of the plurality of Ra substituents is relative to-D-NR₁The E group being in the ortho position or, if D is absent, relative to-NR₁-radicalThe group is located in the ortho position and the other substituent or substituents may be located in any other position(s). In some embodiments, the other substituent or substituents are relative to-D-NR₁The E group being in the ortho and/or para position, or relative to the-NR group if D is absent₁The radicals are in the ortho and/or para position.

According to some embodiments of any of the embodiments described herein, when n is 3, each Ra substituent is halogen, e.g., chloro and/or fluoro, or each is chloro.

In some of these embodiments, the plurality of halogen substituents are relative to-D-NR₁The E group being in the ortho and para positions, or relative to the-NR group if D is absent₁The radicals are in the ortho and para positions.

According to some embodiments of any of the embodiments described herein, n is 3; one of the Ra substituent(s) is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl or heteroalicyclic, and is in relation to-D-NR₁The E group being in the ortho position or, if D is absent, relative to-NR₁-the groups are in the ortho position; and the two other Ra substituents are both halogen, as described herein. In some of these embodiments, the two halogen Ra substituents are relative to-D-NR₁The E group being in the ortho and para positions, or relative to the-NR group if D is absent₁The radicals are in the ortho and para positions.

According to some embodiments of any of the embodiments described herein, n is 3; one of the Ra substituent(s) is alkyl, preferably mono-lower alkyl or mono-cycloalkyl, and is in respect of-D-NR₁The E group being in the ortho position or, if D is absent, relative to-NR₁-the groups are in the ortho position; and the two other Ra substituents are both halogen, as described herein. In some of these embodiments, the two halogen Ra substituents are relative to-D-NR₁The E group being in the ortho and para positions, or relative to the-NR group if D is absent₁-the groups are in ortho-and para-position;

according to what is described hereinIn some embodiments of any of the embodiments (e.g., for formula I or Ia), n is 3; one of the Ra substituent(s) is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl or heteroalicyclic, and is in relation to-D-NR₁The E group being in the ortho position or, if D is absent, relative to-NR₁-the groups are in the ortho position; and the two other Ra substituents are both halogen, as described herein; m is 1; and Rb is halogen (e.g., fluorine), and is relative to-E-NR₁The group-D-is in the para position, or if E is absent, relative to-NR₁The group is in the para position.

According to some embodiments of any of the embodiments described herein (e.g., for formula I or Ia), n is 3; one of the Ra substituent(s) is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl or heteroalicyclic, and is in relation to-D-NR₁The E group being in the ortho position or, if D is absent, relative to-NR₁-the groups are in the ortho position; both of the two other Ra substituents are halogen, as described herein; and m is 1, and the Rb substituent corresponds to-E-NR₁The group-D-is in the para position, or if E is absent, relative to-NR₁The group is in the para position.

According to some embodiments of any of the embodiments described herein (e.g., for formula I or Ia), n is 3; one of the Ra substituent(s) is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl or heteroalicyclic, and is in relation to-D-NR₁The E group being in the ortho position or, if D is absent, relative to-NR₁-the groups are in the ortho position; both of the two other Ra substituents are halogen, as described herein; m is 1; and the Rb substituent is halogen (e.g., fluorine).

According to some embodiments of any of the embodiments described herein (e.g., for formula I or Ia), one of the Ra substituent(s) is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl or heteroalicyclic, preferably alkyl (e.g., lower alkyl) or cycloalkyl, and is in contrast to-D-NR-r₁The E group being in the ortho position or, if D is absent, relative to-NR₁-the groups are in the ortho position; m is 1; and Rb is halogen (e.g., fluorine), and corresponds to-E-NR₁The group-D-is in the para position, or if E is absent, relative to-NR₁The group is in the para position.

According to some embodiments of any of the embodiments described herein (e.g., for formula I or Ia), one of the Ra substituent(s) is alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl or heteroalicyclic, preferably alkyl (e.g., lower alkyl) or cycloalkyl, and is in contrast to-D-NR-r₁The E group being in the ortho position or, if D is absent, relative to-NR₁-the groups are in the ortho position; m is 1, and the Rb substituent corresponds to-E-NR₁The group-D-is in the para position, or if E is absent, relative to-NR₁The group is in the para position.

In some embodiments of any embodiment wherein there are two halo Ra substituents, the two halo Ra substituents are relative to-D-NR₁The E group being in the ortho and para positions, or relative to the-NR group if D is absent₁The radicals are in the ortho and para positions.

According to some embodiments of any of the embodiments described herein, k is 1. Optionally, k is 0.

According to some embodiments of any of the embodiments described herein, R is 0 when k is not₂And R₃Each is hydrogen. Alternatively, R₂And R₃One or both of which is a mono-alkyl or mono-haloalkyl, preferably a mono-lower alkyl.

According to some of any of the embodiments described herein, k is 1, and R is₂And R₃Each is hydrogen.

According to some of any of the embodiments described herein, R is₄Is hydrogen. Alternatively, R₄Is a mono-alkyl group, preferably a lower alkyl group. Further alternatively, R₄And R₅、R₆、R₇And R₈Or with R₅、R₆、R₇、R₈、R₉(if present) and R₁₀One or more of which (if not hydrogen) together form a heterocyclic ring, preferably a heteroalicyclic ring.

According to some embodiments of any of the embodiments described herein, k is 1; r₂And R₃Each is hydrogen; and R₄Is hydrogen.

According to some of any of the embodiments described herein, X is absent such that Z is an alkylene chain or an alicyclic ring (a cycloalkyl) substituted with Y, or a heteroalicyclic ring, wherein Y and optionally NR are₄Forming part of the loop.

According to some of any of the embodiments described herein, X is O, such that Z is a monoalkylene glycol chain terminated by Y, or is a monoalkylene Chain (CR)₅R₆) w is connected to a heteroalicyclic ring formed between X and Y, or to R₇、R₈Are linked (e.g., where q is greater than 1).

According to some of any of the embodiments described herein, X is O and Z is a monoalkylene glycol chain terminated by Y. In some of these embodiments, w is 1,2 or 3, preferably 2, and q is 1,2 or 3, preferably 2.

According to some embodiments of any of the embodiments described herein, X is absent, and Z is represented by (CR)₅R₆) w and (CR)₇R₈) q is an alkylene chain.

According to some embodiments of any of the embodiments described herein, X is absent, and Z is represented by (CR)₅R₆) w and (CR)₇R₈) q form an alicyclic ring (a cycloalkyl group), wherein R₅、R₆、R₇And R₈Two or more of which form the ring, substituted with Y.

According to some of any of the embodiments described herein, the sum of w and q is at least 2, and in some embodiments it is at least 3, for example, 3, 4, 5 or 6 or more.

According to some of any of the embodiments described herein, R is₅、R₆、R₇And R₈At least one of which is independently selected from alkyl, haloalkyl and halogen, and/or R₅、R₆、R₇And R₈At least two of which together form a cycloaliphatic ring.

According to some of any of the embodiments described herein, R is₅、R₆、R₇And R₈At least two of which are independently selected from alkyl, haloalkyl and halogen.

According to some of any of the embodiments described herein, R is₅、R₆、R₇And R₈At least two of which are each independently a monoalkyl group, preferably a lower alkyl group.

According to some of any of the embodiments described herein, q is 1, and R is₇And R₈At least one or each of which is an alkyl group, preferably a lower alkyl group.

According to some embodiments of any of the embodiments described herein, w is 1 or 2.

According to some of any of the embodiments described herein, R is₅And R₆Each is hydrogen.

When w is not 1, it is composed of two or more (CR)₅R₆) Groups, and R in each of these groups₅And R₆May be the same or different.

When q is not 1, it is composed of two or more (CR)₇R₈) Groups, and R in each of these groups₇And R₈May be the same or different.

According to some of any of the embodiments described herein, q is 1, and w is 1 or 2.

According to some of these embodiments, R₇And R₈Is not hydrogen, and is preferably mono-alkyl (e.g. a lower alkyl, such as methyl). Alternatively, or additionally, at the (CR)₅R₆) R in one of the radicals₅And R₆Is not hydrogen, and is preferably monoalkyl (e.g., a lower alkyl, such as methyl).

According to some embodiments of any of the embodiments described herein, q is 1 and w is 2.

According to some embodiments of any of the embodiments described herein, q is 2 and w is 2.

According to some embodiments of any of the embodiments described herein, at (CR)₇R₈) In at least one of the radicals, R₇And R₈Is not hydrogen, and is preferably monoalkyl (e.g., a lower alkyl, such as methyl). Alternatively, or additionally, at the (CR)₅R₆) R in one of the radicals₅And R₆Is not hydrogen, and is preferably monoalkyl (e.g., a lower alkyl, such as methyl).

According to some embodiments of any of the embodiments described herein, X is absent, and R is₅、R₆、R₇And R₈At least two of which together form an alicyclic ring. In some of these embodiments, w is 1 and q is 1, and R is₅、R₆、R₇And R₈Together form the cycloaliphatic ring.

In some of these embodiments, w is 2 and q is 1, and an alicyclic ring is substituted with NR₄Connected (CR)₅R₆) And (4) forming a group. Alternatively, w is 1 and q is 2, and a cycloaliphatic radicalThe ring being formed by a ring not linked to Y (CR)₇R₈) And (4) forming a group.

In some embodiments of any of the embodiments described herein, whenever Z comprises a cycloaliphatic ring, the ring is a three to six membered ring, either a five or six membered ring.

According to some embodiments of any of the embodiments described herein, X is absent, and R is₅、R₆、R₇And R₈At least two of which together form an alicyclic ring (a cycloalkyl group). In some of these embodiments, w and q are each 1, and in some embodiments q is 1 and w is 1 or 2. In some of these embodiments, R₇And R₈Together form an alicyclic ring (a cycloalkyl), and in some other embodiments, R₅、R₆、R₇And R₈All together form an alicyclic ring. The cycloaliphatic ring may have 3, 4, 5, 6 or more carbon atoms.

According to some of any of the embodiments described herein, X is O, and R is₅、R₆、R₇And R₈Preferably R₇And/or R₈Is not hydrogen (e.g., monoalkyl, cycloalkyl, aryl).

According to some of any of the embodiments described herein, X is O, w is 2 or greater, and R is₅And R₆Two of which together form an alicyclic ring.

According to some of any of the embodiments described herein, X is O, q is 2 or greater, and R is₇And R₈Two of which together form an alicyclic ring.

According to some of any of the embodiments described herein, X is NR₉And R is₉And R₅、R₆、R₇And R₈And one or more of the heteroalicyclic rings together.

In some embodiments of any of the embodiments described herein, R is₅、R₆、R₇And R₈At least one of which is independently selected from alkyl, haloalkyl and halogen, and/or R₅、R₆、R₇And R₈At least two of which together form an alicyclic ring and at least one of the Ra substituent(s) is selected from the group consisting of alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl and heteroalicyclic; alternatively, two Ra substituents together form a ring. In some of these embodiments, the Ra substituent is relative to-D-NR, as described herein₁the-E-group is in the ortho position. In some embodiments of any of these embodiments, m is 1 and the Rb substituent is halo, e.g., fluoro, as described herein in the various embodiments.

In some embodiments of any of the embodiments described herein, R is₅、R₆、R₇And R₈At least one of which is independently an alkyl group, and/or R₅、R₆、R₇And R₈At least two of which together form an alicyclic group and one of the Ra substituent(s) is selected from the group consisting of alkyl, haloalkyl, cycloalkyl, aryl, alkoxy, alkylamino, heteroaryl and heteroalicyclic, or, alternatively, two Ra substituents together form a ring. In some of these embodiments, the Ra substituent is relative to-D-NR, as described herein₁the-E-group is in the ortho position. In some embodiments of any of these embodiments, m is 1 and the Rb substituent is halo, e.g., fluoro, as described herein in the various embodiments.

In some embodiments of any of the embodiments described herein, R is₅、R₆、R₇And R₈At least one of which is independently selected from alkyl, haloalkyl and halogen, and/or R₅、R₆、R₇And R₈At least two of which together form an alicyclic ring and at least one of the Ra substituent(s) is selected from alkyl, haloalkyl and cycloalkyl. In some of these embodiments, the Ra substituent is relative to-D-NR, as described herein₁the-E-group is in the ortho position. In some embodiments of any of these embodiments, m is 1 and the Rb substituent is halo, e.g., fluoro, as described herein in the various embodiments.

In some embodiments of any of the embodiments described herein, R is₅、R₆、R₇And R₈At least one of which is independently an alkyl group, and/or R₅、R₆、R₇And R₈At least two of which together form an alicyclic ring and at least one of the Ra substituent(s) is selected from alkyl, haloalkyl and cycloalkyl. In some of these embodiments, the Ra substituent is relative to-D-NR, as described herein₁the-E-group is in the ortho position. In some embodiments of any of these embodiments, m is 1 and the Rb substituent is halo, e.g., fluoro, as described herein in the various embodiments.

According to some embodiments of any of the embodiments described herein, Y is hydroxy, such that Y is OR₁₀And R is₁₀Is hydrogen.

According to some embodiments of any of the embodiments described herein, a compound of formula I or Ia as described herein has a one L ogP value of at least 3, preferably at least 4, for example between 4 and 5, when octanol and water are determined.

A number of exemplary compounds according to this embodiment are shown in fig. 19.

As described herein, exemplary compounds according to this embodiment include compounds: NH66, NH91, NH101, NH110, NH83.1, NH83.2, NH83.3, NH82.1, NH82.2, NH82.3, and NH 160.

As described herein, exemplary compounds according to this embodiment include compounds: NH91, NH101, NH110, NH83.1, NH83.2, NH83.3, NH82.1, NH82.2, and NH 82.3.

As described herein, exemplary compounds according to this embodiment include compounds: NH91, NH101, NH110, NH83.1, NH83.2, and NH 83.3.

As described herein, exemplary compounds according to this embodiment include compounds: NH91, NH101, and NH 110.

As described herein, exemplary compounds according to this embodiment include compounds: NH91 and NH 101.

The compounds of this embodiment can be readily prepared by methods known in the art, generally by coupling one of the starting materials corresponding to ring B and one of the starting materials corresponding to ring A in formula I, using at the appropriate position a linker which can couple to each other and form a-D-NR₁-reactive groups of the E-group.

The coupling may be performed such that the starting material corresponding to ring B already comprises a group V as defined herein, or, alternatively, the starting material may comprise a corresponding ester or carboxylic acid instead of an amide, and after coupling with the starting material corresponding to ring a, a suitable amine is coupled to form an amide in V.

In some embodiments, when both D and E are absent, the starting material corresponding to ring B comprises an amine group at the corresponding position (methyl to V) and the starting material corresponding to ring a comprises a leaving group, as defined herein, which can participate in a nucleophilic coupling reaction.

Exemplary synthetic routes are described in the examples section below.

For any of the embodiments described herein and any combination thereof, the compound may be in the form of a salt, e.g., a pharmaceutically acceptable salt.

As used herein, the term "pharmaceutically acceptable salt" refers to a charged species of the parent compound and its counter ion, which is typically used to modify the solubility characteristics of the parent compound and/or reduce any significant irritation of an organism by the parent compound, while not abrogating the biological activity and properties of the administered compound. A pharmaceutically acceptable salt of a compound as described herein may be formed alternately during the synthesis of the compound, for example, during isolation of the compound from a reaction mixture or recrystallization of the compound.

In the context of some of this embodiments, a pharmaceutically acceptable salt of a compound described herein can optionally be an acid addition salt comprising at least one basic group (e.g., an amine and/or amide and/or a nitrogen atom in a heterocyclyl) of the compound in a positively charged form (e.g., where the basic group is protonated), in combination with at least one counter ion derived from the selected acid, to form a pharmaceutically acceptable salt.

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

The acid addition salts may be mono-addition salts or poly-addition salts depending on the stoichiometric ratio between the charged group(s) in the compound and the counter ion in the salt.

The term "mono-addition salt" as used herein refers to a salt wherein the stoichiometric ratio between the counter ion and the charged form of the compound is 1:1, such that the addition salt comprises 1 molar equivalent of counter ion per molar equivalent of the compound.

The term "poly-addition salt" as used herein refers to a salt wherein the stoichiometric ratio between the counter ion and the charged form of the compound is greater than 1:1, and is, for example, a salt of 2:1, 3:1, 4:1, etc., such that the addition salt includes 2 or more molar equivalents of a counter ion per molar equivalent of the compound.

One non-limiting example of a pharmaceutically acceptable salt can be an ammonium cation or a guanidine cation and acid addition salts thereof.

The acid addition salts can include a variety of organic and inorganic acids such as, but not limited to, hydrochloric acid to provide a hydrochloric acid addition salt, hydrobromic acid to provide a hydrobromic acid addition salt, acetic acid to provide an acetic acid addition salt, ascorbic acid to provide an ascorbic acid addition salt, benzenesulfonic acid to provide a monobenzenesulfonate salt, camphorsulfonic acid to provide a camphorsulfonic acid addition salt, citric acid to provide a citric acid addition salt, maleic acid to provide a maleic acid addition salt, malic acid to provide a malic acid addition salt, methanesulfonic acid, which provides a methanesulfonic acid (mesylate) addition salt, naphthalenesulfonic acid, which provides a naphthalenesulfonic acid addition salt, oxalic acid, which provides an oxalic acid addition salt, phosphoric acid, which provides a monophosphate addition salt, toluenesulfonic acid, which provides a p-toluenesulfonic acid addition salt, succinic acid, which provides a succinic acid addition salt, sulfuric acid, which provides a sulfuric acid addition salt, tartaric acid, which provides a tartaric acid addition salt, and trifluoroacetic acid, which provides a trifluoroacetic acid addition salt. Each of these acid addition salts may be a mono-addition salt or a poly-addition salt, as these terms are defined herein.

Embodiments of the present invention also encompass any enantiomer, diastereomer, prodrug, solvate, hydrate, and/or pharmaceutically acceptable salt of a compound described herein.

As used herein, the term "enantiomer" refers to a stereoisomer of a compound that can only be registered relative to its counterpart by virtue of being completely inverted/reflected (mirrored) with respect to each other. Enantiomers are referred to as having "chirality" because they correspond to each other like the left and right hands. Enantiomers have the same chemical and physical properties except when present in an environment that is itself chiral, such as in all living systems. In the context of this embodiment, a compound may exhibit one or more chiral centers, each exhibiting an R-or S-configuration and any combination, and compounds according to some embodiments of the invention may have any of their chiral centers exhibiting an R-or S-configuration.

The term "diastereomer" as used herein refers to stereoisomers that are not enantiomers of each other. Diastereoisomers occur when two or more stereoisomers of a compound have different configurations at one or more, but not all, of the equivalent (relevant) stereocenters, and are not mirror images of each other. When two diastereomers differ from each other in only one stereocenter, they are epimers. Each stereocenter (chiral center) results in two different configurations, and thus two different stereoisomers. In the context of the present invention, the various embodiments of the present invention encompass compounds having multiple chiral centers, which occur in any combination of stereoconfigurations, i.e. any diastereomer.

The term "prodrug" refers to a formulation that is converted in vivo to the active compound (active parent drug). Prodrugs are commonly used to facilitate the administration of the parent drug. They may, for example: bioavailability is obtained by oral administration, whereas the parent drug is not. Prodrugs may also have improved solubility in pharmaceutical compositions compared to the parent drug. Prodrugs are also often used to achieve sustained release of the active compound in vivo.

The term "solvate" refers to a complex of variable stoichiometry (e.g., two, three, four, five, six, etc.) formed by a solute (a compound of the invention) and a solvent, wherein the solvent does not interfere with the biological activity of the solute. Suitable solvents include, for example, ethanol, acetic acid, and the like.

The term "hydrate" refers to a mono-solvate as defined above, wherein the solvent is water.

According to an aspect of some embodiments of the present invention there is provided a pharmaceutical composition comprising a compound as described herein and a pharmaceutically acceptable carrier.

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 an activity of a voltage-dependent potassium ion channel.

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

In some embodiments, the potassium channel is Kv7.2/7.3.

In some embodiments, the modulating comprises opening the potassium ion channel.

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

In some embodiments, the modulation comprises inhibiting the activity of the TRPV1 channel (blocking the channel).

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-dependent potassium channel and a TRPV1 channel as described herein in any of the respective embodiments.

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

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 the treatment of a medical condition associated with an activity of a voltage-dependent potassium channel and/or a TRPV1 channel.

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 the treatment of a medical condition associated with an activity of a voltage-dependent potassium channel and a TRPV1 channel.

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

According to some embodiments of any of the embodiments described herein, the medical condition is such that modulating an activity of one, and preferably both, of a voltage-dependent potassium channel and a TRPV1 channel as described herein is beneficial.

According to some of any of the embodiments described herein, the medical condition is such that opening a voltage-dependent potassium channel and blocking (inhibiting) a TRPV1 channel as described herein is beneficial.

One exemplary medical condition is neuropathic pain.

Any other medical condition (pathology, disorder, disease and/or disease) associated with the TRPV1 channel function and/or a voltage-dependent potassium channel described herein is contemplated.

Exemplary medical conditions that can be beneficially treated by the TRPV1 inhibitors (blockers) (compounds having the general formula I) described herein include, but are not limited to: epilepsy, pain-related disorders such as neuropathic pain, mild tenderness (allodynia), inflammation-related pain, and pain associated with pancreatitis, bipolar disorder, mood disorders, psychosis, schizophrenia, anxiety, motor neuron disease, overactive bladder, urinary incontinence, persistent visceral hypersensitivity including irritable bowel syndrome, chronic cough, and cancer (squamous cell carcinoma, prostate cancer, and pancreatic cancer).

There are many pathologies, disorders and diseases associated with a defect in potassium channel function. Like other potassium channel opening compounds, the compounds described herein are useful in the framework of treating pathologies, disorders, diseases (diseases) and diseases (disorders) associated with deficient potassium channel function, thereby treating, ameliorating, preventing, inhibiting or limiting the effects of disorders and pathologies on animals, including humans.

Exemplary medical 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, spasticity, mood disorders, brain tumors, psychosis, schizophrenia, muscle twitching (myokymia), neurogenic pain, neuropathic pain, epilepsy (seizure), epilepsy (epilepsy), hearing and vision loss, anxiety and motor neuron disorders. The compounds described herein may further be beneficially used as neuroprotective agents (e.g., to prevent stroke, etc.). The compounds described herein are also useful in the treatment of disease states such as gastroesophageal reflux disease and gastrointestinal motility disorder (gastrointestional hypomotility) disease.

The compounds disclosed herein may also be useful as effective candidates for the treatment of a variety of medical conditions in which decreased cortical and/or peripheral neuronal activity is beneficial, for example: epilepsy, ischemic stroke, migraine, ataxia, muscle twitching (myokymia), neurogenic pain, neuropathic pain, Parkinson's disease, bipolar disorder, trigeminal neuralgia, spasticity, mood disorders, psychosis, schizophrenia, brain tumors, hearing and vision loss, anxiety disorders, and motor neuron disease.

According to an aspect of some embodiments of the invention, the compounds or compositions described herein are useful for inhibiting a cortical and/or peripheral neuron activity, and/or for treating a condition in which inhibition of a cortical and/or peripheral neuron activity in a subject is beneficial, as described herein.

As used herein, a "pharmaceutical composition" refers to a formulation of one or more compounds of the present invention (as active ingredients) or physiologically acceptable salts or prodrugs thereof, with other chemical ingredients, including but not limited to physiologically suitable: carriers, excipients, lubricants, buffers, antibacterial agents, fillers (e.g., mannitol), antioxidants (e.g., ascorbic acid or sodium bisulfite), anti-inflammatory agents, antiviral agents, chemotherapeutic agents, antihistamines, and the like. The purpose of a pharmaceutical composition is to facilitate administration of a compound to a subject.

The term "active ingredient" refers to a compound that is responsible for a biological effect.

The terms "physiologically acceptable carrier" and "pharmaceutically acceptable carrier" are used interchangeably to refer to a carrier or a diluent that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the compound being administered.

The term "excipient" herein refers to a non-acting substance (inert substance) added to a pharmaceutical composition to further facilitate the administration of a drug. Examples of excipients include, but are not limited to: calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils and polyethylene glycols.

Techniques for formulation and administration of drugs are found in the latest version of the "Remington's pharmaceutical Sciences" by Mack Publishing Co, Estan (Easton), Pennsylvania (Pennsylvania, Pa.), which is incorporated herein by reference.

Thus, the pharmaceutical compositions for use in accordance with the present invention may be formulated in conventional manner using one or more pharmaceutically acceptable carriers comprising excipients and auxiliaries, which facilitate processing of the compounds into preparations which can be used pharmaceutically. Appropriate formulation depends on the chosen route of administration. The dosage may vary depending on the dosage form employed and the route of administration employed. The exact prescription, route of administration and dosage can be selected by The individual physician according to The patient's condition (see, e.g., Fingl et al, 1975, "Pharmacological Basis of Therapeutics", Chapter 1, page 1).

The pharmaceutical compositions may be formulated for administration by one or more routes, depending on the choice of local or systemic treatment or administration, and the area to be treated. Administration can be oral, inhalation or parenteral, e.g., intravenous drip or intraperitoneal, subcutaneous, intramuscular or intravenous injection, or topical (including ophthalmic, vaginal, rectal, intranasal) administration.

Formulations for topical administration may include, but are not limited to, detergents, ointments, gels, creams, suppositories, drops, liquid preparations (liquids), sprays, and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable.

Compositions for oral administration include powders or granules, suspensions or solutions in aqueous or non-aqueous media, sachets, pills, caplets, capsules or tablets. Thickeners, diluents, flavoring agents, dispersing aids, emulsifiers or binders may be desirable.

Formulations for parenteral administration may include, but are not limited to, sterile solutions, which may also contain buffers, diluents, and other suitable additives. Sustained release compositions are contemplated for use in therapy.

The amount of a composition administered will, of course, depend on the subject being treated, the severity of the disease, the mode of administration, the judgment of the prescribing physician, and the like.

In some embodiments of the invention, the amount of the composition to be administered (e.g., a dose or a 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 a previously described compound known to exhibit the same therapeutic effect.

If desired, the compositions of the present invention may be provided in a packaging or dispensing device, such as an FDA (food and drug administration) approved kit, which may contain one or more unit dosage forms containing the active ingredient. The pack may, for example, comprise metal or plastic foil, such as but not limited to a blister pack (bib pack) or pressurised container (for inhalation). The packaging or dispensing device may be accompanied by instructions for administration. The packaging or dispensing device may also be accompanied by a notice associated with the container in a format prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals, which notice reflects approval by the agency of the form of the composition or human or veterinary administration. The notification may be, for example, a label for a prescription drug approved by the U.S. food and drug administration, or an insert for an approved product. As detailed above, compositions comprising an SRI of the present invention formulated in a compatible pharmaceutical carrier may also be prepared, placed in a suitable container, and labeled for treatment of a particular medical condition, disease or disorder.

The pharmaceutical composition may also comprise additional pharmaceutically active or inactive agents such as, but not limited to: an antibacterial agent, an antioxidant, a buffering agent, a bulking agent, a surfactant, an anti-inflammatory agent, an antiviral agent, a chemotherapeutic agent, and an antihistamine, and/or an adjunct useful in the treatment of a medical condition, disease or disorder as described herein.

As used herein, the term "about" refers to ± 10% or ± 5%.

The terms "comprising", "including", "having", and variations thereof mean "including, but not limited to".

The term "consisting of" means "including and limited to.

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

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

Throughout this application, various embodiments of the present invention may be presented in a range format. It is to be understood that the description in range format is merely for convenience and brevity and should not be construed as a mandatory limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible sub-ranges as well as individual numerical values within the range. For example, a description of a range from 1 to 6 should be considered to have explicitly disclosed sub-ranges, such as: from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., as well as individual values within the stated ranges, such as 1,2, 3, 4, 5, and 6, for example, apply regardless of the breadth of the ranges.

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

As used herein, the term "method" refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures either known to, or readily developed from known manners, means, techniques and procedures by, practitioners of the chemical, pharmacological, biological, biochemical and medical arts.

As used herein, the term "treating" includes eliminating, substantially inhibiting, slowing or reversing the progression of the condition, substantially ameliorating clinical or aesthetic symptoms of the condition, or substantially preventing the appearance of clinical or aesthetic symptoms of the condition.

As used herein, the term "alkyl" refers to saturated aliphatic hydrocarbons including straight and branched chain groups. Preferably, the alkyl group has 1 to 20 carbon atoms. Every time a numerical range; for example, "1 to 20" as used herein refers to a group, in this case an alkyl group, which may contain 1 carbon atom, 2 carbon atoms3 carbon atoms, and the like, up to and including 20 carbon atoms. More preferably, it is a medium size alkyl group having 1 to 10 carbon atoms. Most preferably, it is a lower alkyl group having 1 to 4 carbon atoms. Alkyl groups may be substituted or unsubstituted. When substituted, the substituent may be, for example, cycloalkyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, halogen, carbonyl, thiocarbonyl, O-carbamoyl (O-carbamyl), N-carbamoyl (N-carbamyl), O-thiocarbamoyl (O-thiocarbamoyl), N-thiocarbamoyl (N-thiocarbamoyl), C-amido (C-amido), N-amido (N-amido), C-carboxy, O-carboxy, nitro, sulfonamide, trihalomethanesulfonamide (trihalomethanesulphonamide), silyl (silylium), formamidine (guanidyl), guanido (guanidino), ureido, amino or NR_aR_bWherein R is_aAnd R_bEach independently hydrogen, alkyl, cycloalkyl, aryl, carbonyl, sulfonyl, trihalomethylsulfonyl (trihalomethylsulfonyl), and five or six membered heterocycloaliphatic rings.

A "haloalkyl" group describes a monoalkyl group as defined herein substituted with one or more halo substituents as defined herein. In some embodiments, haloalkyl is a mono-alkyl substituted with two or more or three or more halo substituents. In some embodiments, each halogen substituent is fluorine. In some embodiments, the monohaloalkyl is-CF₃or-CF₂H。

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 fully conjugated pi-electron system (an alicyclic ring). Examples of cycloalkyl groups include, but are not limited to: cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, cyclohexadiene, cycloheptane, cycloheptatriene, and adamantane. Cycloalkyl groups may be substituted or unsubstituted. When substituted, the substituents can be, for example, alkyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, halogen, carbonyl,Thiocarbonyl, C-carboxy, O-carbamoyl (O-carbamyl), N-carbamoyl (N-carbamyl), C-amido (C-amidi), N-amido (N-amidi), nitro, amino and NR as defined above_aR_b。

"alkenyl" refers to an alkyl group consisting of at least two carbon atoms and at least one carbon-carbon double bond.

"alkynyl" refers to an alkyl group consisting of at least two carbon atoms and at least one carbon-carbon triple bond.

In some embodiments, whenever an alkyl substituent is indicated, it may be substituted with an alkynyl or a monoalkynyl group as defined herein.

An "aryl" group refers to an all-carbon monocyclic or fused-ring polycyclic (i.e., rings which share adjacent pairs of carbon atoms) groups having a fully conjugated pi-electron system. Examples of aryl groups include, but are not limited to: phenyl, naphthyl and anthracenyl. The aryl group may be substituted or unsubstituted. When substituted, the substituents may be, for example: halogen, trihalomethyl, alkyl, hydroxy, alkoxy, aryloxy, thiohydroxy, thiocarbonyl, C-carboxy, O-carbamoyl (O-carbamyl), N-carbamoyl (N-carbamyl), O-thiocarbamoyl (O-thiocarbamoyl), N-thiocarbamoyl (N-thiocarbamoyl), C-amido (C-amidi), N-amido (N-amidi), sulfinyl, sulfonyl, amino and NR as defined above_aR_b。

A "heteroaryl" group refers to a monocyclic or fused ring (i.e., rings that share adjacent pairs of atoms) group that has one or more atoms, such as nitrogen, oxygen, and sulfur, in the ring(s), and also has a fully 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 may be substituted or unsubstituted. When substituted, the substituents may be, for example, alkyl, cycloalkyl, halogen, trihalomethyl, hydroxy, alkoxy, aryloxy, thiohydroxy, thiocarbonyl, sulfonamido, C-carboxy, O-carboxy, sulfinyl, sulfonyl, O-carbamoyl (O-carbamoyl), N-carbamoyl (N-carbamoyl)O-thiocarbamoyl (O-thiocarbamoyl), N-thiocarbamoyl (N-thiocarbamoyl), C-amido (C-amidi), N-amido (N-amidi), amino or NR as defined above_aR_b。

"Heterocyclyl" refers to a monocyclic or fused ring group having one or more atoms in the ring(s), such as nitrogen, oxygen, and sulfur. The rings may also have one or more double bonds. However, these rings do not have a fully conjugated pi-electron system. The heteroalicyclic may be substituted or unsubstituted. When substituted, the substituents may be, for example, alkyl, cycloalkyl, aryl, heteroaryl, halogen, trihalomethyl, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, carbonyl, thiocarbonyl, C-carboxy, O-carbamoyl (O-carbamoyl), N-carbamoyl (N-carbamoyl), O-thiocarbamoyl (O-thiocarbamoyl), N-thiocarbamoyl (N-thiocarbamoyl), sulfinyl, sulfonyl, C-amido (C-amidio), N-amido (N-amidio), amino and NR as defined above_aR_b。

As used herein, "cyclyl" describes an alicyclic group (a cycloalkyl), an aryl, a heteroaryl, or a heteroalicyclic.

"hydroxy" refers to an-OH group.

"azido" refers to the group-N ═ N.

As defined herein, "alkoxy" refers to-O-alkyl and-O-cycloalkyl.

"haloalkoxy" describes an O-alkyl group, wherein the alkyl group is a haloalkyl group as described herein.

As defined herein, "aryloxy" refers to-O-aryl and-O-heteroaryl.

"thiol" or "thiol" refers to the-SH group.

As defined herein, "thioalkoxy" refers to-S-alkyl and-S-cycloalkyl.

As defined herein, "thioaryloxy" refers to-S-aryl and-S-heteroaryl groups.

"carbonyl" 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.

An "aldehyde" group refers to a carbonyl group, wherein R' is hydrogen.

"thiocarbonyl" refers to the group-C (═ S) -R ', where R' is as defined herein.

The term "carboxylate" includes C-carboxylates and O-carboxylates.

"C-carboxy" refers to the group-C (═ O) -O-R ', where R' is as defined herein.

"O-carboxy" refers to an R 'C (═ O) -O-group, where R' is as defined herein.

A "carboxylic acid" group refers to a C-carboxyl group wherein R' is hydrogen.

"halo" group means fluoro, chloro, bromo, or iodo.

"Trihalomethyl" group means-CX₃A group wherein X is a halogen group as defined herein.

"Trihalosulfonyl" means X₃CS(＝O)₂-a group, wherein X is a halogen group as defined herein.

"sulfinyl" refers to the group-S (═ O) -R ', where R' is as defined herein.

"Sulfonyl" means-S (═ O)₂-R 'wherein R' is as defined herein.

The term "sulfonamide group" includes S-sulfonamide and N-sulfonamide groups.

"S-sulfonamido" means-S (═ O)₂-NR ' R "group, wherein R ' is as defined herein and R" is as defined for R '.

"N-sulfonamido" refers to R' S (═ O)₂-NR "group, wherein R' and R" are as defined herein.

"Tri-halogenated methanesulfonamido" means X₃CS(＝O)₂NR '-groups, wherein R' and X are as defined herein.

The term "carbamate" includes O-carbamoyl and N-carbamoyl.

"O-carbamoyl" refers to the group-OC (═ O) -NR 'R ", where R' and R" are as defined herein.

"N-carbamoyl" refers to the group R 'OC (═ O) -NR ″, where R' and R "are as defined herein.

The term "thiocarbamate" includes O-thiocarbamoyl and N-thiocarbamoyl.

"O-thiocarbamoyl" refers to the group-OC (═ S) -NR 'R ", where R' and R" are as defined herein.

"N-thiocarbamoyl" refers to the R "OC (═ S) NR '-group, where R' and R" are as defined herein.

"amino" refers to the group-NR 'R ", wherein R' and R" are as defined herein.

"alkylamino" refers to a monoamino group wherein one of R 'and R "is alkyl (monoalkylamine), or R' and R" are both independently alkyl (dialkylamine).

The term "amide" encompasses C-amide groups and N-amide groups.

"C-amido" refers to the group-C (═ O) -NR 'R ", where R' and R" are as defined herein.

"N-amido" refers to an R 'C (═ O) -NR "group, where R' and R" are as defined herein.

The "quaternary ammonium" group is-NHR' R "⁺Wherein R 'and R' are independently alkyl, cycloalkyl, aryl or heteroaryl.

"ureido" refers to the group-NR ' C (═ O) -NR "R '", where R ' and R "are as defined herein and R '" is defined as R ' or R ".

"guanidino" refers to the group-R 'NC (═ N) -NR "R'", where R ', R ", and R'" are as defined herein.

"carbamimidoyl" refers to a group R 'R "NC (═ N) -, wherein R' and R" are as defined herein.

"nitro" means-NO₂A group.

"cyano" refers to a-C.ident.N group.

"silyl" refers to-SiR 'R "R'", wherein R ', R ", and R'" are as defined herein.

As used herein and in the art, "leaving group" describes a labile atom, group, or chemical moiety that is susceptible to disassociation from an organic molecule during a chemical reaction, typically facilitated by the relative stability of the atom, group, or moiety that is being disassociated therefrom. In general, any group that is the conjugate base of a strong acid may be used as a leaving group. Representative examples of suitable leaving groups according to some embodiments of the present invention include, but are not limited to: trichloroacetimidate, acetate, tosylate, triflate, sulfonate, azide, halide (halogen, preferably bromine or iodine), hydroxy, thiol, alkoxy, cyanate, thiocyanate, nitro and cyano.

It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination or as suitable in any other described embodiment of the invention. Certain features described in the context of various embodiments are not considered essential features of those embodiments, unless the embodiments do not function without those elements.

Various embodiments and aspects of the present invention described above and claimed in the following claims are supported experimentally in the following examples.

Examples of the invention

Reference is now made to the following examples, which together with the above description illustrate some embodiments of the invention in a non-limiting manner.

Example 1

Structure-activity relationship research-chemical synthesis

To screen these compounds for in vitro activity on the recombinant Kv7.2/3 and TRPV1 channels expressed in CHO cells, the inventors designed and synthesized a library of nearly 50 bis (aryl/heteroaryl) amine derivatives.

Compound libraries were designed according to general formula a:

wherein:

a and B are each independently selected from an aryl and a heteroaryl;

d and E are each independently (CRdRe) u, wherein Rd and Re are each independently hydrogen, alkyl, cycloalkyl, haloalkyl, halogen, or the like; and u is independently 0 or 1 (preferably Rd and Re are each hydrogen);

n is an integer of 1 to 5;

m is an integer of 0 to 5;

ra and Rb as defined in formula I or each independently are selected from alkyl, cycloalkyl, halogen, haloalkyl, haloalkoxy, alkoxy, aryl, aryloxy, hydroxy, amine, alkylamine, thiohydroxy, thioalkoxy, thioaryloxy, cyano, carboxylate, amide, carbamate, sulfonyl and sulfonamide (preferably hydrogen, alkyl, halogen, haloalkyl and haloalkoxy), wherein when n is greater than 1, each substituent Ra is the same or different; when m is greater than 2, each substituent Rb is the same or different;

g is selected from NR₁And O;

R₁is hydrogen, alkyl, cycloalkyl or aryl (preferably hydrogen); and

v is (CR)₂R₃)k-C(＝O)-NR₄-Z and is in ortho, meta or para position relative to G, wherein:

k is an integer from 0 to 2;

R₂and R₃Each independently selected from hydrogen, halogen, alkyl, cycloalkyl and aryl (preferably hydrogen or alkyl);

R₄is hydrogen, alkyl, cycloalkyl or aryl (preferably hydrogen, or with R)₅、R₆、R₇And R₈An alkyl group forming a heteroalicyclic ring, as described below); and

z is represented by formula II:

wherein:

w and q are each independently an integer of 0 to 8, 0 to 6, or 0 to 4, provided that w + q is at least 2 (e.g., 2, 3, or 4);

x is selected from O and NR₉Or absent;

y is selected from OR₁₀、SR₁₀And NR₁₀R₁₁；

R₅、R₆、R₇And R₈Each independently selected from hydrogen, halogen, alkyl, cycloalkyl, heteroalicyclic, aryl, alkylamino, alkoxy, and aryloxy (preferably hydrogen or alkyl); or, alternatively, R₅、R₆、R₇、R₈And R₉Two of which together form an alicyclic or heteroalicyclic ring; and

R₁₀and R₁₁Each independently is hydrogen or alkyl; or, alternatively, R₅、R₆、R₇、R₈、R₉、R₁₀And R₁₁Two of which together form an alicyclic or heteroalicyclic ring.

The library is designed so that the screened compounds differ from each other by one or more of the following structural features:

the position of variable V relative to G;

the nature of G;

whether or not D and/or E are present;

the number and nature of the Ra substituent(s) and their positions;

the presence or absence, nature and location of the Rb substituent(s);

the nature of ring A and ring B; and

chemical structure of variable V.

The chemical structures of exemplary compounds in the designed library are listed in table a.

As can be seen, most of the compounds are characterized by the benzene rings of the A and B rings. D and E are absent, G is NR₁And R is₁Is hydrogen. These compounds are also referred to herein as diphenylamine compounds or derivatives.

Fig. 1A-1B illustrate an exemplary general synthetic route for the preparation of the diphenylamine derivatives shown in table a.

In fig. 1A, an exemplary synthetic route begins with coupling of the corresponding amine to a carboxylic acid derivative of phenyl substituted with (Rb) m as defined in formula a, and further with a group NR corresponding to variable V related to formula a₁An exemplary non-limiting reagent is HATU, however, any other coupling reagent or reagent system (e.g., for peptide coupling) can be considered, synthesis using a reducing agent or system to convert the nitro group to an amine an exemplary reduction system includes hydrogen, Pd/C, and optionally an alcoholic solvent, such as methanol, however, any other reducing agent or system suitable for converting the nitro group to an amine can be considered, the synthesis then proceeds to couple the corresponding amine with a benzene ring substituted by (Ra) n as defined by formula (a) and further substituted by a leaving group (represented as L in fig. 1A)₃、Xant-phos、K₂CO₃Dioxane and heat (e.g. microwave heating at 160 ℃), however, any other reagent or reagent system is contemplated.

In fig. 1B, an exemplary synthetic route begins with the esterification of a carboxylic acid derivative of a phenyl group substituted with (Rb) m as defined in formula a, and further by a group NR located corresponding to the variable V associated in formula a₁A nitro group at one position (e.g., ortho or methyl). Any reagent or reagent system for esterification is contemplated. Exemplary esterification is carried out using an acid such as sulfuric acid. The synthesis is carried out by reducing the nitro group to an amine and coupling the resulting amine with a benzene ringAnd said phenyl ring is substituted as defined by formula a with (Ra) n and further with a leaving group as depicted in figure 1A. The resulting compound is then de-esterified using methods and reagents known in the art, and the resulting carboxylic acid is coupled with the corresponding amine to form the amide of variable V of formula a, as depicted in figure 1A.

By selecting starting materials and reagents having certain substituents on the aromatic rings (corresponding to variables Ra and Rb in formula a) and/or amides (corresponding to variable V in formula a), the diphenylamine compounds of formula a are prepared, respectively.

Fig. 2 to 6 show an exemplary synthesis of representative diphenylamine derivatives: NH66 (fig. 2); NH82 and NH82 (fig. 3); NH91 (fig. 4); HN101 (fig. 5) and NH110 (fig. 6).

Using a similar synthetic route, the following compounds can be prepared:

compounds having heteroaryl as one of ring a and ring B in formula a, and/or having variable G in addition to NH and/or variables D and/or E, are prepared by slightly modifying these pathways.

Fig. 7 provides a synthetic route for preparing exemplary compounds having heteroaryl as ring B, represented as NH 106.

Example 2

Study on structure-activity relationship

The compound described in example 1 was tested for dual activity in modulating kv7.2/3 and TRPV1 channels using the following protocol:

chinese hamster ovary CHO cells were grown in DMEM supplemented with 2mM glutamine, 10% fetal bovine serum, and antibiotics. Briefly, 40000 cells seeded on poly-D-lysine coated glass coverslips (13 mm in diameter) in 24-well plates were transfected using pIRES-CD8 (0.3. mu.g) as a marker to transfect 0.5. mu.g of Kv7.2 and 0.5. mu.g of Kv7.3. Transfection was performed using 3.6. mu. l X-tremagene 9(Roche) according to the manufacturer's protocol. For electrophysiology, transfected cells were visualized using the anti-CD 8 antibody-coated magnetic bead method approximately 40 hours after transfection.

For Kv7.2 potassium ion current recording, the patch pipette was pulled from borosilicate glass (Warner Instrument Corp), resistance 3-7 M.OMEGA., and filled (in mM): 130KCl, 5Mg ATP, 5EGTA, 10HEPES, pH7.3 (adjusted using KOH), and sucrose was added to adjust the osmolality to 290 mOsmol. The external solution contained (in mM): 140NaCl, 4KCl, 1.2MgCl₂、1.8CaCl₂11 glucose, 5.5HEPES, pH7.3 (adjusted using NaOH), and sucrose was added to adjust the osmolality to 310 mOsmol. Cells were held at-90 mV and stepped from-70 mV to +30mV in 10mV increments for 1.5 seconds and repolarized at-60 mV. For TRPV1 current recording, the solution was the same as that used for Kv7.2, except that the extracellular solution did not contain CaCl₂To limit desensitization and comprises 1mM MEGTA and 1mM MgCl₂。

After analysis of the data obtained, structural features were identified that made certain di (aryl/heteroaryl) amine derivatives potential drugs with dual channel targeting, activation of kv7.2 and inhibition of TRPV1, as shown below:

an amide substituent of aryl/heteroaryl ring B of formula a (variable V in formula a) in the meta position of the group connecting the two aryl/heteroaryl rings (variable G in formula a);

with alkyl or cycloalkyl functional groups (as R)₅、R₆、R₇And R₈One or more) hydroxyalkyl or alkylene glycol substituents introduced into the amide (variable Z in formula a);

introducing an alkyl or cycloalkyl substituent (as an Ra substituent) in the ortho position relative to the G group of the aryl/heteroaryl ring that does not contain an amide substituent (ring a in formula a); and/or

A halogen (e.g., fluorine) substituent (as an Rb substituent) is introduced in the ortho position to ring B of formula a relative to the amide substituent (variable V in formula a).

In addition, compounds other than the diphenylamide derivatives defined herein are shown, which are characterized by the variables A, B, D, E and the other groups of G in formula A being inferior to the diphenylamine derivatives.

The following compounds were found to exert the best dual targeting effect, activating kv7.2 and inhibiting TRPV1 (see table a): NH66, NH82, NH83, NH91, NH101 and NH110, and further comparative studies were performed.

Example 3

Activity assay

Chinese hamster ovary CHO cells were grown in DMEM supplemented with 2mM glutamine, 10% fetal bovine serum, and antibiotics. Briefly, 40000 cells seeded on poly-D-lysine coated glass coverslips (13 mm in diameter) in 24-well plates were transfected using pIRES-CD8 (0.3. mu.g) as a marker to transfect 0.5. mu.g of Kv7.2 and 0.5. mu.g of Kv7.3. Transfection was performed using 3.6. mu. l X-tremagene 9(Roche) according to the manufacturer's protocol. For electrophysiology, transfected cells were visualized using the anti-CD 8 antibody-coated magnetic bead method approximately 40 hours after transfection. Whole cell configurations using patch clamp technique were recorded. The signal was amplified using an Axomatch 200B patch clamp amplifier (Axon Instruments), sampled at 5kHz, and filtered at 2.4kHz through a 4-pole Bessel (Bessel) low pass filter. Data was acquired using pClamp10.5 software in conjunction with the DigiData 1440A interface. The membrane pipette is pulled from borosilicate glass (Harvard Apparatus) with a resistance of 3-7 megaohms. Intracellular straw solution contained 130mM KCl, 5mM K2-ATP, 5mM EGTA (or 5mM BAPTA when indicated), 10mM HEPES, pH7.3 (adjusted with KOH), CaCl₂(for different free Ca according to MAXCHE L ATOR software²⁺Concentration values required) and sucrose was added to adjust the osmotic pressure to 290 mosmol. The external solution contained 140mM NaCl, 4mM KCl, 1.8mM CaCl₂、1.2mM MgCl₂11mM glucose, 5.5mM HEPES, adjusted to pH7.3 using NaOH (310 mOsM). The series resistance is compensated (75-90%) and monitored periodically. The Clampfit program (pClam10.5; Axon Instruments), Microsoft Excel (Microsoft, Redmond, Washington) and Prism 5 were usedData analysis was performed 0(GraphPad Software, Inc., san Diego, Calif.). The leakage subtraction was performed offline using the Clampfit program of the pClamp10.5 software. Cells were kept at-90 mV and stepped from-70 mV to +30mV in 10mV increments and repolarized at-60 mV.

Normalized conductivity was plotted as a function of test voltage for cells in the absence and presence of drug. The chord conductance (G) is calculated by the following equation: g ═ I/(V-Vrev), where I corresponds to the current amplitude measured at the end of the pulse and Vrev is the calculated reversal potential assumed to be-90 mV in CHO cells. G is estimated at various test voltages (V) and then normalized to a maximum conductance value Gmax. Activation curves were fitted by a Boltzmann distribution: G/Gmax ═ 1/{1+ exp [ (V50-V)/s ] }, where V50 is the voltage at which the current is semi-activated and s is the slope factor.

Inhibition of the recombinant TRPV1 channel was tested as described in example 2 above.

TRPV1 whole cell current was activated by exposing transfected CHO cells to 0.5. mu.M capsaicin at a holding potential of-60 mV.

The current recording of cardiac channel HERG is performed as follows:

cells were maintained at-90 mV and stepped from-70 mV to +30mV in 10mV increments for 1.5 seconds and repolarized at-60 mV.

The inhibition of the spike-firing induced by sensory neurons of the Dorsal Root Ganglion (DRG) which sense pain is tested as follows:

for current clamp recordings in DRG neurons, patch pipettes (patch pipettes) were filled (in mM): 135KCl, 1K₂ATP、1MgATP、2EGTA、1.1CaCl₂5 glucose (free [ Ca ]²⁺]_i87nM), 10HEPES, adjusted with KOH at pH 7.4(315 mOsm). The external solution contained (in mM): 150NaCl, 2.5KCl, 2CaCl₂、2MgCl₂15 glucose, 10HEPES, adjusted with NaOH at pH 7.4(325 mOsm). Drug burst was induced by rapid application using a rapid perfusion system (AutoMate Scientific).

As a result:

FIGS. 8A to 8B show the effect of 5. mu.M NH66 on the amplitude (FIG. 8A) and voltage dependence of the recombined Kv7.2/3 current, and the effect of the aforementioned diphenylamine compound NH43 (FIG. 8C).

As shown therein, NH43 and NH66 increased the amplitude of the recombinant Kv7.2/3 current expressed in CHO cells by about 3.5-fold at-40 mV at 5. mu.M and shifted its voltage dependence to the left by more than-10 mV.

Figure 9 shows the effect of 5 μ M NH66 on recombinant TRPV1 current activated by 0.1 μ M Capsaicin (CAP). FIG. 9 shows TRPV1 current induced by the application of 0.1 μ M capsaicin to transfected CHO cells. The upper part shows the current induced by applying capsaicin only 0.1 μ M; in the middle is shown the current induced first by applying 0.1 μ M capsaicin with 5 μ M NH66, and then only by 0.1 μ M capsaicin. The lower panel shows the current induced first by 0.1 μ M capsaicin, then by 0.1 μ M capsaicin and 5 μ M NH 66.

As shown therein, NH66, like NH43, inhibited capsaicin-activated recombinant TRPV1 current, IC, at 0.1. mu.M₅₀0.3. mu.M.

Fig. 10A-10B show the effect of 1 μ M NH43 (fig. 10A) and 1 μ M NH66 (fig. 10B) on the induced spiking of rat DRG neurons.

As shown therein, NH43 and NH66 inhibit the sensory neuron-induced spiking, IC, of Dorsal Root Ganglion (DRG) sensory neurons that sense pain₅₀0.3. mu.M and 0.5. mu.M, respectively.

FIGS. 11A-11B show the effect of NH82 on the amplitude (FIG. 11A) and voltage dependence (FIG. 11B) of recombination Kv7.2/3.

Fig. 11C-11D show the effect of NH82 (fig. 11C) and NH83 (fig. 11D) on DRG-induced spiking. As shown therein, NH82 affected the recombinant kv7.2/3 and TRPV1 channels as well as DRG-induced spiking, and NH83 showed slightly improved activity on DRG-induced spiking. Among other assays, NH91, NH101, and NH110 were identified as the most potent compounds tested in the recombination channel and DRG-induced spiking.

Fig. 12A to 12B, 13A to 13C, and 14A to 14B show the effect of NH91, NH101, and NH110 on the amplitude (fig. 12A, 13A, and 14A) and voltage dependence (fig. 12B, 13B to 13C, and 14B) at-40 mV for the kv7.2/3 channel, respectively, and show that these compounds effectively increase the amplitude, shifting the voltage dependence of activation to the left.

FIG. 15A shows that NH91 significantly suppressed DRG-induced spiking. FIGS. 15B-15C show that NH91(5 μ M) does not affect inward voltage-dependent Ca in DRG neurons²⁺And Na⁺Current, showing its specificity for Kv7.2/3 and TRPV1 currents.

Fig. 16 shows that NH110 also suppresses DRG-induced spiking.

The dual targeting and synergistic effect of NH91 is further illustrated in fig. 17A-B and fig. 18A-18B by its effect on 1 μ M capsaicin activation of DRG neuronal spiking triggered by the TRPV1 channel.

Fig. 17A and 17B show that NH91 produces a significantly more effective inhibition of capsaicin-induced DRG spikes compared to the known TRPV1 antagonist AMG 517.

Although the TRPV1 antagonist AMG517 (0.1. mu.M, is its IC)₅₀100-fold) inhibited capsaicin-induced DRG spike by 42% (n-11, one-way anova, P)<0.0001), but NH91 produced a significantly more potent inhibition of 88% (n 10, one-way ANOVA, P)<0.0001). These data indicate that NH91 shows a clear synergy in DRG compared to typical high affinity gold standard TRPV1 antagonists (e.g., AMG 517). This suggests a synergistic effect of NH91 in inhibiting TRPV1 channel and NH91 activating kv7.2/3 channel.

Further support for this argument is shown in fig. 18A-18B, namely: NH91 (50%) inhibited capsaicin-induced DRG spiking even in the presence of kv7.2/3 channel blocker XE991(10 μ M), which also showed its potent inhibition of the TRPV1 channel (n ═ 11, two-tailed unpaired t test, P < 0.0001).

Lead compounds were further tested for their affinity for cardiac hERG inhibitory activity (off target) to assess possible adverse effects on cardiac channels.

Table 1 below summarizes the main parameters demonstrated in the in vitro studies. Starting from two initial heads NH29 and NH34 (openers of kv7.2/3 and blockers of TRPV1) that exhibit dual channel targeting, Structure Activity Relationships (SAR) studies based on the present invention have identified a set of potential leads from a rational chemical design of about 50 New Chemical Entities (NCE).

The initial sprouts had lower affinity for both targets (Kv7.2/3 and TRPV1) than the initial sprouts, but had micromolar affinity for cardiac hERG inhibitory activity (off target), the potential lead compounds had significantly higher affinity for the targets (0.2-1 μ M) and very low affinity for cardiac hERG channels (35-135 μ M IC)_50s) Provides good cardiac safety values, improves the efficacy of these compounds, and improves the therapeutic index as a whole.

The optimized NCE has dual-channel targeting property, and can activate Kv7.2 and inhibit TRPV 1.

TABLE 1

ND is not determined

While the invention has been described in conjunction with specific embodiments thereof, it is evident that 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.

All publications, patents, and patent applications mentioned in this specification are herein incorporated in their entirety by reference into the specification, to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated herein by reference. In addition, citation or identification of any reference in this application shall not be construed as an admission that such reference is available as prior art to the present invention. To the extent that a section heading is used, it should not be construed as necessarily limiting.

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