Compounds for the Treatment of Neuromuscular Disorders
20260007625 ยท 2026-01-08
Assignee
Inventors
- Thomas Holm PEDERSEN (Aarhus N, DK)
- Lars J.S. KNUTSEN (Aarhus N, DK)
- Nicholas Michael KELLY (Aarhus N, DK)
- Farman ULLAH (Aarhus N, DK)
- Neerja SARASWAT (Aarhus N, DK)
- Rajesh KUMAR (Aarhus N, DK)
Cpc classification
A61P21/00
HUMAN NECESSITIES
C07C323/62
CHEMISTRY; METALLURGY
A61K31/192
HUMAN NECESSITIES
C07C323/53
CHEMISTRY; METALLURGY
International classification
A61K31/192
HUMAN NECESSITIES
A61P21/00
HUMAN NECESSITIES
C07C323/53
CHEMISTRY; METALLURGY
C07C323/62
CHEMISTRY; METALLURGY
Abstract
The present disclosure relates to compounds suitable for treating, ameliorating and/or preventing neuromuscular disorders, including the reversal of drug-induced neuromuscular blockade. The compounds as defined herein can inhibit the CIC-1 ion channel.
Claims
1. A compound of Formula (I): ##STR00025## wherein: R.sup.1 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; OC.sub.2-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; R.sup.2 is selected from the group consisting of C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 and C.sub.2-3 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.8 or when R.sup.1 is OC.sub.2-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 then R.sup.2 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 and C.sub.2-3 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.8; R.sup.3 is selected from the group consisting of H and F; R.sup.4 is selected from the group consisting of F, Cl, Br and I; R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; R.sup.6 is selected from the group consisting of H; C.sub.1-5 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkynyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.3-6 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; phenyl optionally substituted with one or more, identical or different, substituents R.sup.9; and benzyl optionally substituted with one or more, identical or different, substituents R.sup.9; R.sup.7 is independently selected from the group consisting of deuterium, F and Cl; R.sup.8 is independently selected from the group consisting of deuterium, F, Cl, OC.sub.1-5 alkyl, SC.sub.1-5 alkyl, N(O)C.sub.1-5 alkyl and N(C.sub.1-5 alkyl)-C(O)C.sub.1-5 alkyl; and R.sup.9 is independently selected from the group consisting of deuterium, methoxy, nitro, cyano, Cl, Br, I, and F; or a pharmaceutically acceptable salt, hydrate, polymorph, tautomer, or solvate thereof with the proviso that when R.sup.1 is F or Me, R.sup.2 is Me, R.sup.3 is H, R.sup.4 is Cl or Br and R.sup.5 is OMe, then R.sup.6 is not H, Me or Et; and with the proviso that when R.sup.1 is Cl or Br, R.sup.2 is Me, R.sup.3 is H, R.sup.4 is F and R.sup.5 is OMe, then R.sup.6 is not H, Me or Et; and with the proviso that when R.sup.1 is F, R.sup.2 is Me, R.sup.3 is F, R.sup.4 is F and R.sup.5 is OMe, then R.sup.6 is not H.
2. The compound according to claim 1, wherein R.sup.1 is Me.
3. The compound according to claim 1, wherein R.sup.2 is Me.
4. The compound according to claim 1, wherein R.sup.3 is H.
5. The compound according to claim 1, wherein R.sup.4 is F or Cl.
6. The compound according to claim 1, wherein R.sup.5 is selected from the group consisting of OMe, OCH.sub.2F, OCHF.sub.2, OCF.sub.3 and SMe.
7. The compound according to claim 1, wherein R.sup.6 is H.
8. The compound according to claim 1, wherein the compound is selected from the group consisting of: 3-chloro-2-cyclopropoxy-5,6-dimethylbenzoic acid; 2,5-dichloro-6-methoxy-3-methylbenzoic acid; 3-chloro-5,6-dimethyl-2-(methylsulphanyl)benzoic acid; 3-chloro-5,6-dimethyl-2-(trifluoromethoxy)benzoic acid; 3-chloro-2-(difluoromethoxy)-5,6-dimethylbenzoic acid; 3-chloro-2-(fluoromethoxy)-5,6-dimethylbenzoic acid; 3-fluoro-2-methoxy-5,6-dimethylbenzoic acid; and 3-chloro-2-ethoxy-5,6-dimethylbenzoic acid.
9. A composition comprising the compound according to claim 1 and a pharmaceutically acceptable carrier.
10. (canceled)
11. A method for treating a disease comprising administering the compound according to claim 1 to a person in need thereof, wherein the disease is selected from the group consisting of myasthenia gravis, autoimmune myasthenia gravis, congenital myasthenic syndrome, seronegative myasthenia gravis, muscle specific kinase myasthenia gravis (MuSK-MG), Lambert-Eaton Syndrome, critical illness myopathy, amyotrophic lateral sclerosis (ALS), spinal muscular atrophy (SMA), critical illness myopathy (CIM), Charcot-Marie Tooth disease, diabetic polyneuropathy, periodic paralysis, hypokalemic periodic paralysis, hyperkalemic periodic paralysis, myotubular myopathy, Duchenne muscular dystrophy, Guillain-Barre syndrome, poliomyelitis, post-polio syndrome, chronic fatigue syndrome, critical illness polyneuropathy, metabolic myopathy, Kennedy's disorder, multiple sclerosis, sarcopenia and multifocal motor neuropathy.
12. The method according to claim 11, wherein the disease is sarcopenia.
13. The method according to claim 11, wherein the administering of the compound comprises reversing and/or ameliorating a neuromuscular blockade.
14. A compound of Formula (I): ##STR00026## wherein: R.sup.1 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; R.sup.2 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 and C.sub.2-3 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.8; R.sup.3 is selected from the group consisting of H and F; R.sup.4 is selected from the group consisting of F, Cl, Br and I; R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; R.sup.6 is selected from the group consisting of H; C.sub.1-5 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkynyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.3-6 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; phenyl optionally substituted with one or more, identical or different, substituents R.sup.9; and benzyl optionally substituted with one or more, identical or different, substituents R.sup.9; R.sup.7 is independently selected from the group consisting of deuterium, F and Cl; R.sup.8 is independently selected from the group consisting of deuterium, F, Cl, OC.sub.1-5 alkyl, SC.sub.1-5 alkyl, N(O)C.sub.1-5 alkyl and N(C.sub.1-5 alkyl)-C(O)C.sub.1-5 alkyl; and R.sup.9 is independently selected from the group consisting of deuterium, methoxy, nitro, cyano, Cl, Br, I, and F; or a pharmaceutically acceptable salt, hydrate, polymorph, tautomer, or solvate thereof for use in treating, ameliorating and/or preventing a neuromuscular disorder, and/or for use in reversing and/or ameliorating a neuromuscular blockade.
15. The compound for use according to claim 14, wherein the compound is selected from the group consisting of: 3-bromo-5-fluoro-2,6-dimethoxybenzoic acid; 3-fluoro-2,6-dimethoxy-5-methylbenzoic acid; 5-chloro-2-methyl-3-[(methylsulphanyl)methyl]benzoic acid; 3,5-dichloro-2,6-dimethoxybenzoic acid; 3-chloro-2-cyclopropoxy-5,6-dimethylbenzoic acid; 2,5-dichloro-6-methoxy-3-methylbenzoic acid; 3-chloro-2,6-dimethoxy-5-methylbenzoic acid; 3-chloro-5,6-dimethyl-2-(methylsulphanyl)benzoic acid; 3-chloro-5,6-dimethyl-2-(trifluoromethoxy)benzoic acid; 3-chloro-2-(difluoromethoxy)-5,6-dimethylbenzoic acid; 3-chloro-2-(fluoromethoxy)-5,6-dimethylbenzoic acid; 3-fluoro-2-methoxy-5,6-dimethylbenzoic acid; 3-chloro-2-ethoxy-5,6-dimethylbenzoic acid; 3-bromo-2-methoxy-5,6-dimethylbenzoic acid; and 3-chloro-2-methoxy-5,6-dimethylbenzoic acid.
16. The method of claim 11, wherein the compound is administered as a composition containing the compound and a pharmaceutically acceptable carrier.
17. The method of claim 12, wherein the compound is administered as a composition containing the compound and a pharmaceutically acceptable carrier.
18. The method of claim 13, wherein the compound is administered as a composition containing the compound and a pharmaceutically acceptable carrier.
Description
DETAILED DESCRIPTION
Compounds
[0034] It is within the scope of the present disclosure to provide a compound for use in treating, ameliorating and/or preventing neuromuscular disorders that reduce neuromuscular function. As disclosed herein, inhibition of CIC-1 improves or restores neuromuscular function. The compounds of the present disclosure comprise compounds capable of inhibiting the CIC-1 channel thereby improving or restoring neuromuscular function.
[0035] In one aspect, the disclosure concerns a compound of Formula (I):
##STR00002## [0036] wherein: [0037] R.sup.1 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; OC.sub.2-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0038] R.sup.2 is selected from the group consisting of C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 and C.sub.2-3 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.8 or when R.sup.1 is OC.sub.2-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 then R.sup.2 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 and C.sub.2-3 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.8; [0039] R.sup.3 is selected from the group consisting of H and F; [0040] R.sup.4 is selected from the group consisting of F, Cl, Br and I; [0041] R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0042] R.sup.6 is selected from the group consisting of H; C.sub.1-5 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkynyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.3-6 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; phenyl optionally substituted with one or more, identical or different, substituents R.sup.9; and benzyl optionally substituted with one or more, identical or different, substituents R.sup.9; [0043] R.sup.7 is independently selected from the group consisting of deuterium, F and Cl; [0044] R.sup.8 is independently selected from the group consisting of deuterium, F, Cl, OC.sub.1-5 alkyl, SC.sub.1-5 alkyl, N(O)C.sub.1-5 alkyl and N(C.sub.1-5 alkyl)-C(O)C.sub.1-5 alkyl; and [0045] R.sup.9 is independently selected from the group consisting of deuterium, methoxy, nitro, cyano, Cl, Br, I, and F; [0046] or a pharmaceutically acceptable salt, hydrate, polymorph, tautomer, or solvate thereof with the proviso that when R.sup.1 is F or Me, R.sup.2 is Me, R.sup.3 is H, R.sup.4 is Cl or Br and R.sup.5 is OMe, then R.sup.6 is not H, Me or Et; and with the proviso that when R.sup.1 is Cl or Br, R.sup.2 is Me, R.sup.3 is H, R.sup.4 is F and R.sup.5 is OMe, then R.sup.6 is not H, Me or Et; and with the proviso that when R.sup.1 is F, R.sup.2 is Me, R.sup.3 is F, R.sup.4 is F and R.sup.5 is OMe, then R.sup.6 is not H.
[0047] In one embodiment, R.sup.1 is F. In one embodiment, R.sup.1 is C.sub.1. In one embodiment, R.sup.1 is Br. In one embodiment, R.sup.1 is I. In one embodiment, R.sup.1 is C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8. In one embodiment, R.sup.1 is OC.sub.2-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7. In one embodiment, R.sup.1 is OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7. In one embodiment, R.sup.1 is SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7. In one embodiment, R.sup.1 is SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7.
[0048] In one embodiment, R.sup.1 is selected from the group consisting of F, Cl, Br, and C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8. In one embodiment, R.sup.1 is selected from the group consisting of Et, Me or Cl. In one embodiment, R.sup.1 is Me.
[0049] In one embodiment, R.sup.2 is C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8. In one embodiment, R.sup.2 is Me, Et or Pr. In one embodiment, R.sup.2 is Me. In one embodiment, R.sup.2 is C.sub.2-3 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.8. In one embodiment, R.sup.2 is F. In one embodiment, R.sup.2 is Cl. In one embodiment, R.sup.2 is Br. In one embodiment, R.sup.2 is I.
[0050] In one embodiment, R.sup.3 is H. In one embodiment, R.sup.3 is F.
[0051] In one embodiment, R.sup.4 is selected from the group consisting of F, Cl and Br. In one embodiment, R.sup.4 is selected from the group consisting of F and Cl. In one embodiment, R.sup.4 is Cl. In one embodiment, R.sup.4 is F. In one embodiment, R.sup.4 is Br.
[0052] In one embodiment, R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7. In one embodiment, R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7.
[0053] In one embodiment, R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7. In one embodiment, R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7. In one embodiment, R.sup.5 is selected from the group consisting of SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7. In one embodiment, R.sup.5 is selected from the group consisting of OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7. In one embodiment, R.sup.5 is selected from the group consisting of SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7. In one embodiment, R.sup.5 is selected from the group consisting of cyclopropoxy, OEt, OMe, OCH.sub.2F, OCHF.sub.2, OCF.sub.3, SMe, SCH.sub.2F, SCHF.sub.2 and SCF.sub.3. In one embodiment, R.sup.5 is selected from the group consisting of OMe, OCH.sub.2F, OCHF.sub.2, OCF.sub.3 and SMe.
[0054] In one embodiment, R.sup.6 is H.
[0055] In one embodiment, R.sup.7 is D. In one embodiment, R.sup.7 is F. In one embodiment, R.sup.7 is Cl.
[0056] In one embodiment, R.sup.8 is D. In one embodiment, R.sup.8 is F. In one embodiment, R.sup.8 is Cl.
[0057] In one embodiment, R.sup.8 is SMe. In one embodiment, R.sup.8 is OMe.
[0058] In one embodiment, the EC.sub.50 of the compound is <10 M, such as <5 M, such as <2 M, and such as <1 M.
[0059] In one aspect, the disclosure concerns a compound of Formula (I):
##STR00003## [0060] wherein: [0061] R.sup.1 is selected from the group consisting of F; Cl; Br and C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; [0062] R.sup.2 is selected from the group consisting of C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; [0063] R.sup.3 is H; [0064] R.sup.4 is selected from the group consisting of F, Cl and Br; [0065] R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; and SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0066] R.sup.6 is H; [0067] R.sup.7 is F; and [0068] R.sup.8 is independently selected from the group consisting of deuterium, F, Cl, OC.sub.1-5 alkyl, SC.sub.1-5 alkyl, N(O)C.sub.1-5 alkyl and N(C.sub.1-5 alkyl)-C(O)C.sub.1-5 alkyl; [0069] or a pharmaceutically acceptable salt, hydrate, polymorph, tautomer, or solvate thereof with the proviso that when R.sup.1 is F or Me, R.sup.2 is Me, R.sup.3 is H, R.sup.4 is Cl or Br and R.sup.5 is OMe, then R.sup.6 is not H; and with the proviso that when R.sup.1 is Cl or Br, R.sup.2 is Me, R.sup.3 is H, R.sup.4 is F and R.sup.5 is OMe, then R.sup.6 is not H.
[0070] In one aspect, the compound is selected from the group consisting of: [0071] 3-chloro-2-cyclopropoxy-5,6-dimethylbenzoic acid; [0072] 2,5-dichloro-6-methoxy-3-methylbenzoic acid; [0073] 3-chloro-5,6-dimethyl-2-(methylsulphanyl)benzoic acid; [0074] 3-chloro-5,6-dimethyl-2-(trifluoromethoxy)benzoic acid; [0075] 3-chloro-2-(difluoromethoxy)-5,6-dimethylbenzoic acid; [0076] 3-chloro-2-(fluoromethoxy)-5,6-dimethylbenzoic acid; [0077] 3-fluoro-2-methoxy-5,6-dimethylbenzoic acid; and [0078] 3-chloro-2-ethoxy-5,6-dimethylbenzoic acid.
[0079] In one embodiment, the compound or the compound for use according to the present disclosure has been modified in order to increase its half-life when administered to a patient, in particular its plasma half-life.
[0080] In one embodiment, the compound or the compound for use according to the present disclosure further comprises a moiety conjugated to said compound, thus generating a moiety-conjugated compound. In one embodiment, said moiety-conjugated compound has a plasma and/or serum half-life being longer than the plasma and/or serum half-life of the non-moiety conjugated compound.
[0081] In one embodiment, the moiety conjugated to the compound or compound for use according to the present disclosure, is one or more type(s) of moieties selected from the group consisting of albumin, fatty acids, polyethylene glycol (PEG), acylation groups, antibodies and antibody fragments.
Neuromuscular Disorders
[0082] The compound or compound for use of the present disclosure may be used for treating, ameliorating and/or preventing a neuromuscular disorder, or reversing neuromuscular blockade.
[0083] The inventors of the present disclosure have shown that inhibition of CIC-1 channels strengthens neuromuscular transmission. CIC-1 function may therefore contribute to muscle weakness in conditions of compromised neuromuscular transmission.
[0084] Thus, in one embodiment of the present disclosure, the compound or the compound for use as described herein inhibits CIC-1 channels. Thus, it is appreciated that compounds and/or compounds for use of Formula (I) inhibit CIC-1 channels.
[0085] The neuromuscular disorder may also include neuromuscular dysfunctions.
[0086] Neuromuscular disorders include for example disorders with symptoms of muscle weakness and fatigue. Such disorders may include conditions with reduced neuromuscular transmission safety factor. In one embodiment the neuromuscular disorders are motor neuron disorders. Motor neuron disorders are disorders with reduced safety in the neuromuscular transmission. In one embodiment motor neuron disorders are selected from the group consisting of amyotrophic lateral sclerosis (ALS) (Killian J M, Wilfong A A, Burnett L, Appel S H, Boland D. Decremental motor responses to repetitive nerve stimulation in ALS. Muscle Nerve, 1994, 17, 747-754), spinal muscular atrophy (SMA) (Wadman R I, Vrancken A F, van den Berg L H, van der Pol W L. Dysfunction of the neuromuscular junction in spinal muscular atrophy types 2 and 3. Neurology, 2012, 79, 2050-2055), Charcot-Marie Tooth disease (Bansagi B, Griffin H, Whittaker R G, Antoniadi T, Evangelista T, Miller J, Greenslade M, Forester N, Duff J, Bradshaw A, Kleinle S, Boczonadi V, Steele H, Ramesh V, Franko E, Pyle A, Lochmller H, Chinnery P F, Horvath R. Genetic heterogeneity of motor neuropathies. Neurology, 2017, 28; 88 (13): 1226-1234), X-linked spinal and bulbar muscular atrophy (Yamada, M., Inaba, A., Shiojiri, T. X-linked spinal and bulbar muscular atrophy with myasthenic symptoms. Journal of the Neurological Sciences, 1997, 146, 183-185), Kennedy's disorder (Stevic, Z., Peric, S., Pavlovic, S., Basta, I., Lavrnic, D., Myasthenic symptoms in a patient with Kennedy's disorder. Acta Neurologica Belgica, 2014, 114, 71-73), multifocal motor neuropathy (Roberts, M., Willison, H. J., Vincent, A., Newsom-Davis, J. Multifocal motor neuropathy human sera block distal motor nerve conduction in mice. Ann Neurol. 1995, 38, 111-118), Guillain-Barr syndrome (Ansar, V., Valadi, N. Guillain-Barr Syndrome Prim. Care, 2015, 42, 189-193); poliomyelitis (Trojan, D. A., Gendron, D., Cashman, N. R. Electrophysiology and electrodiagnosis of the post-polio motor unit. Orthopedics, 1991, 14, 1353-1361, and Birk T. J. Poliomyelitis and the post-polio syndrome: exercise capacities and adaptationcurrent research, future directions, and widespread applicability. Med. Sci. Sports Exerc., 1993, 25, 466-472), post-polio syndrome (Garcia, C. C., Potian, J. G., Hognason, K., Thyagarajan, B., Sultatos, L. G., Souayah, N., Routh, V. H., McArdle, J. J. Acetylcholinesterase deficiency contributes to neuromuscular junction dysfunction in type 1 diabetic neuropathy. Am. J. Physiol. Endocrinol. Metab., 2012, 15, E551-561) and sarcopenia (Gilmore K. J., Morat T., Doherty T. J., Rice C. L., Motor unit number estimation and neuromuscular fidelity in 3 stages of sarcopenia. 2017 55(5): 676-684). In one embodiment, the neuromuscular disorder is diabetic polyneuropathy. In one embodiment, the neuromuscular disorder is sarcopenia. In one embodiment, the neuromuscular disorder is Kennedy's disorder. In one embodiment, the neuromuscular disorder is multifocal motor neuropathy.
[0087] Thus, in one embodiment of the present disclosure the neuromuscular disorder is amyotrophic lateral sclerosis (ALS). In another embodiment the neuromuscular disorder is spinal muscular atrophy (SMA). In another embodiment the neuromuscular disorder is Charcot-Marie tooth disease (CMT). In another embodiment the neuromuscular disorder is sarcopenia. In yet another embodiment, the neuromuscular disorder is critical illness myopathy (CIM).
[0088] As stated above the neuromuscular disorders include for example disorders with symptoms of muscle weakness and fatigue. Such disorder may for example include diabetes (Am. J. Physiol. Endocrinol. Metab., 2012, 15, E551-561).
[0089] In another embodiment the neuromuscular disorders is chronic fatigue syndrome. Chronic fatigue syndrome (CFS) (Fletcher, S. N., Kennedy, D. D., Ghosh, I. R., Misra, V. P., Kiff, K., Coakley, J. H., Hinds, C. J. Persistent neuromuscular and neurophysiologic abnormalities in long-term survivors of prolonged critical illness. Crit. Care Med. 2003, 31, 1012-1016) is the common name for a medical condition characterized by debilitating symptoms, including fatigue that lasts for a minimum of six months in adults. CFS may also be referred to as systemic exertion intolerance disorder (SEID), myalgic encephalomyelitis (ME), post-viral fatigue syndrome (PVFS), chronic fatigue immune dysfunction syndrome (CFIDS), or by several other terms. Symptoms of CFS include malaise after exertion; unrefreshing sleep, widespread muscle and joint pain, physical exhaustion, and muscle weakness.
[0090] In a further embodiment the neuromuscular disorder is a critical illness polyneuropathy (Angelini C. Spectrum of metabolic myopathies. Biochim. Biophys. Acta., 2015, 1852, 615-621) or CIM (Latronico, N., Bolton, C. F. Critical illness polyneuropathy and myopathy: a major cause of muscle weakness and paralysis. Lancet Neurol. 2011, 10, 931-941). Critical illness polyneuropathy and CIM are overlapping syndromes of widespread muscle weakness and neurological dysfunction developing in critically ill patients.
[0091] The neuromuscular disorder may also include metabolic myopathy (Milone, M., Wong, L. J. Diagnosis of mitochondrial myopathies. Mol. Genet. Metab., 2013, 110, 35-41) and mitochondrial myopathy (Srivastava, A., Hunter, J. M. Reversal of neuromuscular block. Br. J. Anaesth. 2009, 103, 115-129). Metabolic myopathies result from defects in biochemical metabolism that primarily affects muscle. These may include glycogen storage disorders, lipid storage disorder and 3-phosphocreatine stores disorder. Mitochondrial myopathy is a type of myopathy associated with mitochondrial disorder. Symptoms of mitochondrial myopathies include muscular and neurological problems such as muscle weakness, exercise intolerance, hearing loss and trouble with balance and coordination. Thus, in one embodiment of the present disclosure the neuromuscular disorder is metabolic myopathy.
[0092] In another embodiment the neuromuscular disorder is periodic paralysis, in particular hypokalemic periodic paralysis which is a disorder of skeletal muscle excitability that presents with recurrent episodes of weakness, often triggered by exercise, stress, or carbohydrate-rich meals (Wu, F., Mi, W., Cannon, S. C., Neurology, 2013, 80, 1110-1116 and Suetterlin, K. et at, Current Opinion Neurology, 2014, 27, 583-590) or hyperkalemic periodic paralysis which is an inherited autosomal dominant disorder that affects sodium channels in muscle cells and the ability to regulate potassium levels in the blood (Ammat, T. et al, Journal of General Physiology, 2015, 146, 509-525).
[0093] In an embodiment the neuromuscular disorder is a myasthenic condition. Myasthenic conditions are characterized by muscle weakness and neuromuscular transmission failure. Congenital myasthenic syndrome (Finlayson, S., Beeson, D., Palace, J. Congenital myasthenic syndromes: an update. Pract. Neurol., 2013, 13, 80-91) is an inherited neuromuscular disorder caused by defects of several types at the neuromuscular junction.
[0094] Myasthenia gravis and Lambert-Eaton syndrome (Titulaer M. J., Lang B., Verschuuren J J. Lambert-Eaton myasthenic syndrome: from clinical characteristics to therapeutic strategies. Lancet Neurol. 2011, 10, 1098-107) are examples of myasthenic conditions. Myasthenia gravis is either an autoimmune or congenital neuromuscular disorder that leads to fluctuating muscle weakness and fatigue. In the most common cases, muscle weakness is caused by circulating antibodies that block acetylcholine (ACh) receptors at the postsynaptic neuromuscular junction, inhibiting the excitatory effects of the neurotransmitter ACh on nicotinic ACh-receptors at neuromuscular junctions (Gilhus, N. E., Owe, J. F., Hoff, J. M., Romi, F., Skeie, G. O., Aarli, J. A. Myasthenia Gravis: A Review of Available Treatment Approaches, Autoimmune Diseases, 2011, Article ID 84739). Lambert-Eaton myasthenic syndrome (also known as LEMS, Lambert-Eaton syndrome, or Eaton-Lambert syndrome) is a rare autoimmune disorder that is characterized by muscle weakness of the limbs. It is the result of an autoimmune reaction in which antibodies are formed against presynaptic voltage-gated calcium channels, and likely other nerve terminal proteins, in the neuromuscular junction. Thirty to fifty percent of patients with acetylcholine receptor (AChR) antibody-negative myasthenia gravis (MG) have antibodies to muscle specific kinase (MuSK) and are referred to as having MuSK-MG (Borges, L. S., Richman, D. P., Muscle-Specific Kinase Myasthenia Gravis, Frontiers in Immunology, 2020, 11, 707). In one embodiment of the present disclosure the neuromuscular disorder is myasthenia gravis. In another embodiment the neuromuscular disorder is autoimmune myasthenia gravis. In another embodiment the neuromuscular disorder is MuSK-MG. In another embodiment the neuromuscular disorder is Lambert-Eaton syndrome. In another embodiment the neuromuscular disorder is seronegative myasthenia gravis.
[0095] X-linked myotubular myopathy is a part of a group of centronuclear myopathies where cell nuclei are abnormally located in the centre of muscle cells instead of their normal location at the periphery. It is one of the severest congenital muscle diseases and is characterised by marked muscle weakness, hypotonia and feeding and breathing difficulties (Dowling J J, Lawlor M W, Das S. X-Linked Myotubular Myopathy. 2002 Feb. 25 [Updated 2018 Aug. 23]. In: Adam M P, Ardinger H H, Pagon R A, et al., editors. GeneReviews [Internet]. Seattle (WA): University of Washington, Seattle; 1993-2022. Available from: https://www.ncbi.nlm.nih.gov/books/NBK1432/). In one embodiment the neuromuscular disorder is myotubular myopathy.
[0096] Duchenne muscular dystrophy is a severe type of muscular dystrophy that primarily affects boys resulting initially in fatigue and muscle weakness (Angelini C, Tasca E, Fatigue in muscular dystrophies, Neuromuscular Disorders, 2012, 22 Suppl 3: S214-20). In one embodiment the neuromuscular disorder is Duchenne muscular dystrophy.
[0097] Multiple sclerosis (MS) is the most common demyelinating disease, in which the insulating covers of nerve cells in the brain and spinal cord are damaged. This damage disrupts the ability of parts of the nervous system to transmit signals, resulting in a range of signs and symptoms, including physical, mental, and sometimes psychiatric problems. It has been shown that people with multiple sclerosis typically experience greater levels of exercise-induced fatigue compared with healthy individuals (Brotherton et al, J. Neuorophysiol. 2022, 128, 105-117). There are four types of MS in what is known as the Lublin classification: clinically isolated syndrome (CIS), relapsing-remitting MS (RRMS), primary progressive MS (PPMS) and secondary progressive MS (SPMS). In one embodiment, the neuromuscular disorder is selected from the group consisting of multiple sclerosis (MS), clinically isolated syndrome (CIS), relapsing-remitting MS (RRMS), primary progressive MS (PPMS) and secondary progressive MS (SPMS).
[0098] Neuromuscular blockade is used in connection with surgery under general anaesthesia. Reversing agents are used for more rapid and safer recovery of muscle function after such blockade. Complications with excessive muscle weakness after blockade during surgery can result in delayed weaning from mechanical ventilation and respiratory complications after the surgery. These complications can have pronounced effects on outcome of the surgery and future quality of life of patients, there is a need for improved reversing agents (Murphy G. S., Brull S. J. Residual neuromuscular block: lessons unlearned. Part I: definitions, incidence, and adverse physiologic effects of residual neuromuscular block. Anaesth Analg. 2010, 111, 120-128). Thus, in one embodiment, the neuromuscular disorder has been induced by a neuromuscular blocking agent. In one particular embodiment the neuromuscular disorder is muscle weakness caused by neuromuscular blockade after surgery. In another embodiment of the present disclosure the compound or the compound for use is used for reversing and/or ameliorating neuromuscular blockade after surgery. In one embodiment, the neuromuscular blockade is drug induced. In one embodiment, the neuromuscular blockade is caused by non-depolarizing neuromuscular blocker or antibiotic agent. In one embodiment the neuromuscular blockade is induced by an antibiotic. In one embodiment the neuromuscular blockade is induced by a non-depolarizing neuromuscular blocker.
[0099] In one embodiment the compound or the compound for use of the present disclosure is used to prevent a neuromuscular disorder. The compound or the compound for use may for example be used prophylactically against nerve gas that is known to cause symptoms of muscle weakness and fatigue (Kawamura, Y., Kihara, M., Nishimoto, K., Taki, M. Efficacy of a half dose of oral pyridostigmine in the treatment of chronic fatigue syndrome: three case reports. Pathophysiol., 2003, 9, 189-194). In one embodiment the compound or the compound for use of the present disclosure may be used in the treatment of muscle weakness (such as drooping eyelids, loss of facial expression, constipation, muscle weakness in arms, muscle weakness in legs and dyspnoea) caused by botulism poisoning. In one embodiment the compound or the compound for use of the present disclosure may be used in the treatment of snake bites where the snake toxin, such as -neurotoxin or myotoxin, is known to cause symptoms of muscle weakness and fatigue.
[0100] In one embodiment, the neuromuscular disorder is selected from the group consisting of myasthenia gravis, autoimmune myasthenia gravis, congenital myasthenic syndrome, seronegative myasthenia gravis, muscle specific kinase myasthenia gravis (MuSK-MG), Lambert-Eaton Syndrome, critical illness myopathy, amyotrophic lateral sclerosis (ALS), spinal muscular atrophy (SMA), critical illness myopathy (CIM), Charcot-Marie Tooth disease, diabetic polyneuropathy, periodic paralysis, hypokalemic periodic paralysis, hyperkalemic periodic paralysis, myotubular myopathy, Duchenne muscular dystrophy, Guillain-Barre syndrome, poliomyelitis, post-polio syndrome, chronic fatigue syndrome, critical illness polyneuropathy, metabolic myopathy, Kennedy's disorder, multiple sclerosis and multifocal motor neuropathy.
Pharmaceutical Formulations
[0101] In one embodiment, a composition comprising the compound or the compound for use, according to the present disclosure, is provided. The composition according to the present disclosure may be used for treating, ameliorating and/or preventing a neuromuscular disorder, and/or for use in reversing and/or ameliorating a neuromuscular blockade. Thus, the compositions and compounds described herein can be pharmaceutically acceptable. In one embodiment the composition as described herein is in the form of a pharmaceutical formulation. In one embodiment, the composition as described herein further comprises a pharmaceutically acceptable carrier. In one aspect, the present invention concerns a composition comprising the compound as defined herein and a pharmaceutically acceptable carrier.
Combination Therapy
[0102] The composition of the present disclosure may comprise further active ingredients/agents or other components to increase the efficiency of the composition. Thus, in one embodiment the composition further comprises at least one further active agent. It is appreciated that the active agent can be suitable for treating, preventing or ameliorating said neuromuscular disorder.
[0103] The active agent in certain embodiments can be an acetylcholine esterase inhibitor. Said acetylcholine esterase inhibitor may for example be selected from the group consisting of delta-9-tetrahydrocannabinol, carbamates, physostigmine, neostigmine, pyridostigmine, ambenonium, demecarium, rivastigmine, phenanthrene derivatives, galantamine, piperidines, donepezil, tacrine, edrophonium, huperzine, ladostigil, ungeremine and lactucopicrin.
[0104] In certain embodiments, the acetylcholine esterase inhibitor is selected from the group consisting of neostigmine, physostigmine and pyridostigmine. In certain embodiments, the acetylcholine esterase inhibitor is neostigmine or pyridostigmine.
[0105] The active agent may also be an immunosuppressive drug. Immunosuppressive drugs are drugs that suppress or reduce the strength of the body's immune system. Immunosuppressive drugs include but are not limited to glucocorticoids, corticosteroids, cytostatics, antibodies and drugs acting on immunophilins. In one embodiment the active agent is prednisone.
[0106] The active agent may also be an agent that is used in anti-myotonic treatment. Such agents include for example blockers of voltage gated Na.sup.+ channels, and aminoglycosides.
[0107] The active agent may also be an agent for reversing a neuromuscular blockade after surgery. Such agents include for example neostigmine or sugammadex (Org 25969, tradename Bridion).
[0108] The active agent may also be an agent for increasing ACh release by blocking voltage-gated K.sup.+ channels in the pre-synaptic terminal. Such agent includes 3,4-diaminopyridine (Amifampridine; tradename Firdapse).
[0109] The active agent may also be an agent for increasing the levels of survival motor neuron (SMN) protein that are produced. For example, by alternating the splicing of the SMN2 gene in order to increase the expression of full-length SMN protein from SMN2 (Zanetta C, Nizzardo M, Simone C, Monguzzi E, Bresolin N, Comi G P, Corti S, Molecular therapeutic strategies for spinal muscular atrophies: current and future clinical trials. Clinical Therapeutics, 2014, 36, 128-140). Such agents include antisense oligonucleotides such as Nusinersen (tradename Spinraza) or small molecules such as Risdiplam (tradename Evrysdi).
[0110] The active agent may be a gene therapy, for example by using viral vectors to deliver the SMN1 transgene to the affected motor neurons, where it leads to an increase in SMN protein production. Such gene therapies include onasemnogene abeparvovec (tradename Zolgensma). Such gene therapies include nusinersen (tradename Spinraza), risdiplam (tradename Evrysdi) and Branaplam.
[0111] The active agent may be a small molecule that increases expression of the SMN2 gene, thus increasing the amount of full-length SMN protein available. Such therapies include salbutamol (also, called albuterol; tradename Ventolin),
[0112] The active agent may also be an agent for increasing muscle reactivity. Such agents include skeletal troponin activators such as Tirasemtiv and Reldesemtiv (CK-2127107) (Hwee, D. T., Kennedy, A. R., Hartman, J. J., Ryans, J., Durham, N., Malik, F. I., Jasper, J. R. The small-molecule fast skeletal troponin activator, CK-2127107, improves exercise tolerance in a rat model of heart failure. J. Pharmacol. Exp. Ther., 2015, 353, 159-168). Such agents may also be antibodies that block the activation of the skeletal muscle protein myostatin, such as Apitegromab (SRK-015) or GYM329 (RO7204239),
[0113] The active agent may also be an agent that disrupts or blocks the IgG-FcRn interaction thereby reducing the overall IgG recycling. Such agents may be antibodies, such as the aglycosylated immunoglobulin (Ig)G1 monoclonal antibody Nipocalimab, or IgG1 Fc fragment such as Efgartigimod alfa (tradename Vyvgart).
[0114] The active agent may also be an agent that is an inhibitor of the complement component C5a. The active agent may also be an agent that downregulates the overexpression of PMP22 protein, leading to improvement of neuronal signalling in dysfunctional peripheral nerves. Such agents may be combination drugs such as PXT3003. Alternatively, the active agent may also be an agent that binds to the protein complement component 5 (C5) and inhibits its cleavage into C5a and C5b. Such agents may be Zilucoplan (RA101495).
Methods
[0115] In one aspect, the present invention relates to a compound as defined herein for use as a medicament.
[0116] In one aspect, the present disclosure relates to a compound of Formula (I):
##STR00004## [0117] wherein: [0118] R.sup.1 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0119] R.sup.2 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 and C.sub.2-3 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.8; [0120] R.sup.3 is selected from the group consisting of H and F; [0121] R.sup.4 is selected from the group consisting of F, Cl, Br and I; [0122] R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0123] R.sup.6 is selected from the group consisting of H; C.sub.1-5 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkynyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.3-6 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; phenyl optionally substituted with one or more, identical or different, substituents R.sup.9; and benzyl optionally substituted with one or more, identical or different, substituents R.sup.9; [0124] R.sup.7 is independently selected from the group consisting of deuterium, F and Cl; [0125] R.sup.8 is independently selected from the group consisting of deuterium, F, Cl, OC.sub.1-5 alkyl, SC.sub.1-5 alkyl, N(O)C.sub.1-5 alkyl and N(C.sub.1-5 alkyl)-C(O)C.sub.1-5 alkyl; and [0126] R.sup.9 is independently selected from the group consisting of deuterium, methoxy, nitro, cyano, Cl, Br, I, and F; [0127] or a pharmaceutically acceptable salt, hydrate, polymorph, tautomer, or solvate thereof for use in treating, ameliorating and/or preventing a neuromuscular disorder, and/or for use in reversing and/or ameliorating a neuromuscular blockade.
[0128] In one embodiment, R.sup.1 is F. In one embodiment, R.sup.1 is Cl. In one embodiment, R.sup.1 is Br. In one embodiment, R.sup.1 is I. In one embodiment, R.sup.1 is C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8. In one embodiment, R.sup.1 is OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7. In one embodiment, R.sup.1 is OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7. In one embodiment, R.sup.1 is SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7. In one embodiment, R.sup.1 is SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7.
[0129] In one embodiment, R.sup.1 is selected from the group consisting of F; Cl; Br; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 and OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7. In one embodiment, R.sup.1 is selected from the group consisting of F, Cl, Br, C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8. In one embodiment, R.sup.1 is selected from the group consisting of MeO, Et, Me or Cl. In one embodiment, R.sup.1 is selected from the group consisting of Et, Me or Cl. In one embodiment, R.sup.1 is Me.
[0130] In one embodiment, if R.sup.1 is F; Cl; Br or I then R.sup.2 is not F; Cl; Br or I. In one embodiment, R.sup.2 is C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8. In one embodiment, R.sup.2 is Cl or Me. In one embodiment, R.sup.2 is Me. In one embodiment, R.sup.2 is C.sub.2-3 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.8. In one embodiment, R.sup.2 is F. In one embodiment, R.sup.2 is Cl. In one embodiment, R.sup.2 is Br. In one embodiment, R.sup.2 is I. In one embodiment, R.sup.3 is H. In one embodiment, R.sup.3 is F.
[0131] In one embodiment, R.sup.4 is selected from the group consisting of F, Cl and Br. In one embodiment, R.sup.4 is selected from the group consisting of F and Cl. In one embodiment, R.sup.4 is Cl. In one embodiment, R.sup.4 is F. In one embodiment, R.sup.4 is Br.
[0132] In one embodiment, R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7. In one embodiment, R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7.
[0133] In one embodiment, R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7. In one embodiment, R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7. In one embodiment, R.sup.5 is selected from the group consisting of SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7. In one embodiment, R.sup.5 is selected from the group consisting of OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7. In one embodiment, R.sup.5 is selected from the group consisting of SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7. In one embodiment, R.sup.5 is selected from the group consisting of cyclopropoxy, OEt, OMe, OCH.sub.2F, OCHF.sub.2, OCF.sub.3, SMe, SCH.sub.2F, SCHF.sub.2 and SCF.sub.3. In one embodiment, R.sup.5 is selected from the group consisting of OMe, OCH.sub.2F, OCHF.sub.2, OCF.sub.3 and SMe.
[0134] In one embodiment, R.sup.6 is H.
[0135] In one embodiment, R.sup.7 is F. In one embodiment, R.sup.7 is D. In one embodiment, R.sup.7 is Cl.
[0136] In one embodiment, R.sup.8 is D. In one embodiment, R.sup.8 is F. In one embodiment, R.sup.8 is Cl.
[0137] In one embodiment, R.sup.8 is SMe. In one embodiment, R.sup.8 is OMe.
[0138] In one embodiment, the EC.sub.50 of the compound is <10 M, such as <5 M, such as <2 M, and such as <1 M.
[0139] In one aspect, the disclosure concerns a compound of Formula (I):
##STR00005## [0140] wherein: [0141] R.sup.1 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; OC.sub.2-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0142] R.sup.2 is selected from the group consisting of C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 and C.sub.2-3 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.8 or when R.sup.1 is OC.sub.2-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 then R.sup.2 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 and C.sub.2-3 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.8; [0143] R.sup.3 is selected from the group consisting of H and F; [0144] R.sup.4 is selected from the group consisting of F, Cl, Br and I; [0145] R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0146] R.sup.6 is selected from the group consisting of H; C.sub.1-5 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkynyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.3-6 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; phenyl optionally substituted with one or more, identical or different, substituents R.sup.9; and benzyl optionally substituted with one or more, identical or different, substituents R.sup.9; [0147] R.sup.7 is independently selected from the group consisting of deuterium, F and Cl; [0148] R.sup.8 is independently selected from the group consisting of deuterium, F, Cl, OC.sub.1-5 alkyl, SC.sub.1-5 alkyl, N(O)C.sub.1-5 alkyl and N(C.sub.1-5 alkyl)-C(O)C.sub.1-5 alkyl; and [0149] R.sup.9 is independently selected from the group consisting of deuterium, methoxy, nitro, cyano, Cl, Br, I, and F; [0150] or a pharmaceutically acceptable salt, hydrate, polymorph, tautomer, or solvate thereof for use in treating, ameliorating and/or preventing a neuromuscular disorder, and/or for use in reversing and/or ameliorating a neuromuscular blockade.
[0151] In one embodiment, when R.sup.1 is F or Me, R.sup.2 is Me, R.sup.3 is H, R.sup.4 is Cl or Br and R.sup.5 is OMe, then R.sup.6 is not H, Me or Et. In one embodiment, when R.sup.1 is Cl or Br, R.sup.2 is Me, R.sup.3 is H, R.sup.4 is F and R.sup.5 is OMe, then R.sup.6 is not H, Me or Et. In one embodiment, when R.sup.1 is F, R.sup.2 is Me, R.sup.3 is F, R.sup.4 is F and R.sup.5 is OMe, then R.sup.6 is not H.
[0152] In one aspect, the disclosure concerns a compound of Formula (I):
##STR00006## [0153] wherein: [0154] R.sup.1 is selected from the group consisting of F; Cl; Br; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; [0155] R.sup.2 is selected from the group consisting of C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; [0156] R.sup.3 is H; [0157] R.sup.4 is selected from the group consisting of F, Cl and Br; [0158] R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0159] R.sup.6 is H; [0160] R.sup.7 is F; and [0161] R.sup.8 is independently selected from the group consisting of deuterium, F, Cl, OC.sub.1-5 alkyl, SC.sub.1-5 alkyl, N(O)C.sub.1-5 alkyl and N(C.sub.1-5 alkyl)-C(O)C.sub.1-5 alkyl; [0162] or a pharmaceutically acceptable salt, hydrate, polymorph, tautomer, or solvate thereof for use in treating, ameliorating and/or preventing a neuromuscular disorder, and/or for use in reversing and/or ameliorating a neuromuscular blockade. [0163] In one aspect, when R.sup.1 is F or Me, R.sup.2 is Me, R.sup.3 is H, R.sup.4 is Cl or Br and R.sup.5 is OMe, then R.sup.6 is not H. In one aspect, when R.sup.1 is Cl or Br, R.sup.2 is Me, R.sup.3 is H, R.sup.4 is F and R.sup.5 is OMe, then R.sup.6 is not H.
[0164] In one aspect, the compound for use in treating, ameliorating and/or preventing a neuromuscular disorder, and/or for use in reversing and/or ameliorating a neuromuscular blockade is selected from the group consisting of: [0165] 3-bromo-5-fluoro-2,6-dimethoxybenzoic acid; [0166] 3-fluoro-2,6-dimethoxy-5-methylbenzoic acid; [0167] 5-chloro-2-methyl-3-[(methylsulphanyl)methyl]benzoic acid; [0168] 3,5-dichloro-2,6-dimethoxybenzoic acid; [0169] 3-chloro-2-cyclopropoxy-5,6-dimethylbenzoic acid; [0170] 2,5-dichloro-6-methoxy-3-methylbenzoic acid; [0171] 3-chloro-2,6-dimethoxy-5-methylbenzoic acid; [0172] 3-chloro-5,6-dimethyl-2-(methylsulphanyl)benzoic acid; [0173] 3-chloro-5,6-dimethyl-2-(trifluoromethoxy)benzoic acid; [0174] 3-chloro-2-(difluoromethoxy)-5,6-dimethylbenzoic acid; [0175] 3-chloro-2-(fluoromethoxy)-5,6-dimethylbenzoic acid; [0176] 3-fluoro-2-methoxy-5,6-dimethylbenzoic acid; [0177] 3-chloro-2-ethoxy-5,6-dimethylbenzoic acid; [0178] 3-bromo-2-methoxy-5,6-dimethylbenzoic acid; and [0179] 3-chloro-2-methoxy-5,6-dimethylbenzoic acid.
[0180] In one aspect, the present disclosure relates to a method of treating, preventing and/or ameliorating a neuromuscular disorder, said method comprising administering a therapeutically effective amount of the compound or the compound for use as defined herein to a person in need thereof.
[0181] In one aspect, the present disclosure relates to a method of reversing and/or ameliorating a neuromuscular blockade, said method comprising administering a therapeutically effective amount of the compound or the compound for use as defined herein to a person in need thereof.
[0182] In one aspect, the present disclosure relates to a method for treating and/or ameliorating a neuromuscular disorder, or for reversing and/or ameliorating a neuromuscular blockade comprising administering to a patient in need thereof a therapeutically effective amount of a compound of Formula (I):
##STR00007## [0183] wherein: [0184] R.sup.1 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0185] R.sup.2 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 and C.sub.2-3 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.8; [0186] R.sup.3 is selected from the group consisting of H and F; [0187] R.sup.4 is selected from the group consisting of F, Cl, Br and I; [0188] R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0189] R.sup.6 is selected from the group consisting of H; C.sub.1-5 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkynyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.3-6 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; phenyl optionally substituted with one or more, identical or different, substituents R.sup.9; and benzyl optionally substituted with one or more, identical or different, substituents R.sup.9; [0190] R.sup.7 is independently selected from the group consisting of deuterium, F and Cl; [0191] R.sup.8 is independently selected from the group consisting of deuterium, F, Cl, OC.sub.1-5 alkyl, SC.sub.1-5 alkyl, N(O)C.sub.1-5 alkyl and N(C.sub.1-5 alkyl)-C(O)C.sub.1-5 alkyl; and [0192] R.sup.9 is independently selected from the group consisting of deuterium, methoxy, nitro, cyano, Cl, Br, I, and F;
[0193] or a pharmaceutically acceptable salt, hydrate, polymorph, tautomer, or solvate thereof.
[0194] In one aspect, the present invention related to a compound as defined herein for use in the treatment of an indication selected from the group consisting of myasthenia gravis, autoimmune myasthenia gravis, congenital myasthenic syndrome, seronegative myasthenia gravis, muscle specific kinase myasthenia gravis (MuSK-MG), Lambert-Eaton Syndrome, critical illness myopathy, amyotrophic lateral sclerosis (ALS), spinal muscular atrophy (SMA), critical illness myopathy (CIM), Charcot-Marie Tooth disease, diabetic polyneuropathy, periodic paralysis, hypokalemic periodic paralysis, hyperkalemic periodic paralysis, myotubular myopathy, Duchenne muscular dystrophy, Guillain-Barre syndrome, poliomyelitis, post-polio syndrome, chronic fatigue syndrome, critical illness polyneuropathy, metabolic myopathy, Kennedy's disorder, multiple sclerosis and multifocal motor neuropathy.
[0195] In one aspect, the present disclosure relates to a method for recovery of neuromuscular transmission, said method comprising administering a therapeutically effective amount of the compound or the compound for use as defined herein to a person in need thereof.
[0196] The person in need thereof may be a person having a neuromuscular disorder or a person at risk of developing a neuromuscular disorder or a person having symptoms of muscle weakness and/or fatigue. In another embodiment the person in need thereof is a person with reduced neuromuscular transmission safety with prolonged recovery after neuromuscular blockade. Types of neuromuscular disorders are defined herein above. In an embodiment the person has amyotrophic lateral sclerosis, spinal muscular atrophy, myasthenia gravis or Lambert-Eaton syndrome.
[0197] A therapeutically effective amount is an amount that produces a therapeutic response or desired effect in the person taking it. Administration routes, formulations and dosages can be optimized by persons of skill in the art.
[0198] The method of treatment may be combined with other methods that are known to treat, prevent and/or ameliorate neuromuscular disorders. The treatment method may for example be combined with administration of any of the agents mentioned herein above. In one embodiment the treatment is combined with administration of acetylcholine esterase inhibitor such as for example neostigmine or pyridostigmine.
[0199] In one aspect, the invention relates to a method for recovery of force in muscles with neuromuscular dysfunction, said method comprising administering a compound or a composition as defined herein to a subject in need thereof. The term recovery of force in muscles with neuromuscular dysfunction as used herein refers to the ability of a compound to recover contractile force in nerve-stimulated healthy rat muscle after exposure to submaximal concentration (115 nM) of tubocurarine for 90 mins. Recovery of force is quantified as the percentage of the force prior to tubocurarine that is recovered after addition of the compound. In one embodiment, said recovery of force is >5%, such as >10%, such as >15%, such as >20%, such as >25%, such as >30%, such as >35%.
[0200] In one aspect, the present disclosure relates use of a compound of Formula (I):
##STR00008## [0201] wherein: [0202] R.sup.1 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0203] R.sup.2 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 and C.sub.2-3 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.8; [0204] R.sup.3 is selected from the group consisting of H and F; [0205] R.sup.4 is selected from the group consisting of F, Cl, Br and I; [0206] R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0207] R.sup.6 is selected from the group consisting of H; C.sub.1-5 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkynyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.3-6 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; phenyl optionally substituted with one or more, identical or different, substituents R.sup.9; and benzyl optionally substituted with one or more, identical or different, substituents R.sup.9; [0208] R.sup.7 is independently selected from the group consisting of deuterium, F and Cl; [0209] R.sup.8 is independently selected from the group consisting of deuterium, F, Cl, OC.sub.1-5 alkyl, SC.sub.1-5 alkyl, N(O)C.sub.1-5 alkyl and N(C.sub.1-5 alkyl)-C(O)C.sub.1-5 alkyl; and [0210] R.sup.9 is independently selected from the group consisting of deuterium, methoxy, nitro, cyano, Cl, Br, I, and F;
[0211] or a pharmaceutically acceptable salt, hydrate, polymorph, tautomer, or solvate thereof for the manufacture of a medicament for the treatment, amelioration and/or prevention of a neuromuscular disorder, and/or for reversing and/or ameliorating a neuromuscular blockade.
[0212] One aspect of the disclosure relates to use of a compound as defined herein, for the manufacture of a medicament for the treatment, prevention and/or amelioration of a neuromuscular disorder.
[0213] Another aspect relates to use of a compound as defined herein, for the manufacture of a medicament or a reversal agent for reversing and/or ameliorating a neuromuscular blockade after surgery.
Method of Manufacturing
[0214] In one aspect, the present disclosure relates to methods of manufacturing compounds or compounds for use according to formula (I).
[0215] Compounds according to the present invention may be prepared according to any conventional methods of chemical synthesis known by the skilled person, e.g. those described in the working examples. The starting materials for the processes described in the present application are known or may readily be prepared by conventional methods known by the skilled artisan from commercially available chemicals.
[0216] The end products of the reactions described herein may be isolated by conventional technique such as extraction, crystallisation, distillation, chromatography etc.
[0217] The compounds of this invention may exist in unsolvated as well as in solvated forms with pharmaceutically acceptable solvents such as water, ethanol and the like. In general, the solvated forms are considered equivalent to the unsolvated forms for the purposes of this invention.
Items
[0218] 1. A compound of Formula (I):
##STR00009## [0219] wherein: [0220] R.sup.1 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; OC.sub.2-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0221] R.sup.2 is selected from the group consisting of C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 and C.sub.2-3 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.8 or when R.sup.1 is OC.sub.2-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 then R.sup.2 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 and C.sub.2-3 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.8; [0222] R.sup.3 is selected from the group consisting of H and F; [0223] R.sup.4 is selected from the group consisting of F, Cl, Br and I; [0224] R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0225] R.sup.6 is selected from the group consisting of H; C.sub.1-5 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkynyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.3-6 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; phenyl optionally substituted with one or more, identical or different, substituents R.sup.9; and benzyl optionally substituted with one or more, identical or different, substituents R.sup.9; [0226] R.sup.7 is independently selected from the group consisting of deuterium, F and Cl; [0227] R.sup.8 is independently selected from the group consisting of deuterium, F, Cl, OC.sub.1-5 alkyl, SC.sub.1-5 alkyl, N(O)C.sub.1-5 alkyl and N(C.sub.1-5 alkyl)-C(O)C.sub.1-5 alkyl; and [0228] R.sup.9 is independently selected from the group consisting of deuterium, methoxy, nitro, cyano, Cl, Br, I, and F; [0229] or a pharmaceutically acceptable salt, hydrate, polymorph, tautomer, or solvate thereof with the proviso that when R.sup.1 is F or Me, R.sup.2 is Me, R.sup.3 is H, R.sup.4 is Cl or Br and R.sup.5 is OMe, then R.sup.6 is not H, Me or Et; and with the proviso that when R.sup.1 is Cl or Br, R.sup.2 is Me, R.sup.3 is H, R.sup.4 is F and R.sup.5 is OMe, then R.sup.6 is not H, Me or Et; and with the proviso that when R.sup.1 is F, R.sup.2 is Me, R.sup.3 is F, R.sup.4 is F and R.sup.5 is OMe, then R.sup.6 is not H. [0230] 2. The compound according to any one of the preceding items, wherein R.sup.1 is selected from the group consisting of F, Cl, Br, and C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8. [0231] 3. The compound according to any one of the preceding items, wherein R.sup.1 is selected from the group consisting of Et, Me or Cl. [0232] 4. The compound according to any one of the preceding items, wherein R.sup.1 is Me. [0233] 5. The compound according to any one of the preceding items, wherein R.sup.2 is C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8. [0234] 6. The compound according to any one of the preceding items, wherein R.sup.2 is Me. [0235] 7. The compound according to any one of the preceding items, wherein R.sup.3 is H. [0236] 8. The compound according to any one of the preceding items, wherein R.sup.4 is selected from the group consisting of F, Cl and Br. [0237] 9. The compound according to any one of the preceding items, wherein R.sup.4 is selected from the group consisting of F and Cl. [0238] 10. The compound according to any one of the preceding items, wherein R.sup.4 is Cl. [0239] 11. The compound according to any one of the preceding items, wherein R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7. [0240] 12. The compound according to any one of the preceding items, wherein R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7. [0241] 13. The compound according to any one of the preceding items, wherein R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7. [0242] 14. The compound according to any one of the preceding items, wherein R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7. [0243] 15. The compound according to any one of the preceding items, wherein R.sup.5 is selected from the group consisting of cyclopropoxy, OEt, OMe, OCH.sub.2F, OCHF.sub.2, OCF.sub.3, SMe, SCH.sub.2F, SCHF.sub.2 and SCF.sub.3. [0244] 16. The compound according to any one of the preceding items, wherein R.sup.5 is selected from the group consisting of OMe, OCH.sub.2F, OCHF.sub.2, OCF.sub.3 and SMe. [0245] 17. The compound according to any one of the preceding items, wherein R.sup.6 is H. [0246] 18. The compound according to any one of the preceding items, wherein R.sup.7 is F. [0247] 19. The compound according to any one of the preceding items, wherein R.sup.8 is SMe. [0248] 20. The compound according to any one of the preceding items, wherein the EC.sub.50 of the compound is <10 M, such as <5 M, such as <2 M, and such as <1 M. [0249] 21. The compound according to any one of the preceding items, wherein [0250] R.sup.1 is selected from the group consisting of F; Cl; Br and C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; [0251] R.sup.2 is selected from the group consisting of C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; [0252] R.sup.3 is H; [0253] R.sup.4 is selected from the group consisting of F, Cl and Br; [0254] R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; and SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0255] R.sup.6 is H; [0256] R.sup.7 is F; and [0257] R.sup.8 is independently selected from the group consisting of deuterium, F, Cl, OC.sub.1-5 alkyl, SC.sub.1-5 alkyl, N(O)C.sub.1-5 alkyl and N(C.sub.1-5 alkyl)-C(O)C.sub.1-5 alkyl; [0258] or a pharmaceutically acceptable salt, hydrate, polymorph, tautomer, or solvate thereof with the proviso that when R.sup.1 is F or Me, R.sup.2 is Me, R.sup.3 is H, R.sup.4 is Cl or Br and R.sup.5 is OMe, then R.sup.6 is not H; and with the proviso that when R.sup.1 is Cl or Br, R.sup.2 is Me, R.sup.3 is H, R.sup.4 is F and R.sup.5 is OMe, then R.sup.6 is not H. [0259] 22. The compound according to any one of the preceding items, wherein the compound is selected from the group consisting of: [0260] 3-chloro-2-cyclopropoxy-5,6-dimethylbenzoic acid; [0261] 2,5-dichloro-6-methoxy-3-methylbenzoic acid; [0262] 3-chloro-5,6-dimethyl-2-(methylsulphanyl)benzoic acid; [0263] 3-chloro-5,6-dimethyl-2-(trifluoromethoxy)benzoic acid; [0264] 3-chloro-2-(difluoromethoxy)-5,6-dimethylbenzoic acid; [0265] 3-chloro-2-(fluoromethoxy)-5,6-dimethylbenzoic acid; [0266] 3-fluoro-2-methoxy-5,6-dimethylbenzoic acid; and [0267] 3-chloro-2-ethoxy-5,6-dimethylbenzoic acid. [0268] 23. The compound according to any one of the preceding items, wherein the compound has activity on the CIC-1 receptor. [0269] 24. The compound according to any one of the preceding items, wherein the compound is an inhibitor of the CIC-1 ion channel. [0270] 25. The compound according to any one of the preceding items, wherein the recovery of force in muscles with neuromuscular dysfunction is >5%, for example >10%, for example >15%, for example >20%, for example >25%, for example >30% and for example >35%. [0271] 26. The compound according to any one of the preceding items, wherein the compound improves the recovered force in isolated rat soleus muscles after exposure to tubocurarine. [0272] 27. The compound according to any one of the preceding items for use as a medicament. [0273] 28. A composition comprising the compound according to any one of the preceding items. [0274] 29. The composition according to any one of the preceding items, wherein the composition further comprises a pharmaceutically acceptable carrier. [0275] 30. The composition according to any one of items 28 to 29, wherein the composition further comprises at least one further active agent. [0276] 31. The composition according to item 30, wherein said further active agent is suitable for treating, preventing or ameliorating said neuromuscular disorder. [0277] 32. The compound according to any one of items 1 to 27 or composition according to any one of items 28 to 31 for use in the treatment of an indication selected from the group consisting of myasthenia gravis, autoimmune myasthenia gravis, congenital myasthenic syndrome, seronegative myasthenia gravis, muscle specific kinase myasthenia gravis (MuSK-MG), Lambert-Eaton Syndrome, critical illness myopathy, amyotrophic lateral sclerosis (ALS), spinal muscular atrophy (SMA), critical illness myopathy (CIM), Charcot-Marie Tooth disease, diabetic polyneuropathy, periodic paralysis, hypokalemic periodic paralysis, hyperkalemic periodic paralysis, myotubular myopathy, Duchenne muscular dystrophy, Guillain-Barre syndrome, poliomyelitis, post-polio syndrome, chronic fatigue syndrome, critical illness polyneuropathy, metabolic myopathy, Kennedy's disorder and multifocal motor neuropathy. [0278] 33. The compound according to any one of items 1 to 27 or composition according to any one of items 28 to 31 for use in the treatment of sarcopenia. [0279] 34. The compound according to any one of items 1 to 27 or composition according to any one of items 28 to 31 for use in reversing and/or ameliorating a neuromuscular blockade. [0280] 35. The compound according to any one of items 1 to 27 or composition according to any one of items 28 to 31 for use in the treatment of botulism poisoning, in the treatment of snake bites, in the treatment of nerve gas poisoning or prophylactically against nerve gas poisoning. [0281] 36. A compound of Formula (I):
##STR00010## [0282] wherein: [0283] R.sup.1 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0284] R.sup.2 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 and C.sub.2-3 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.8; [0285] R.sup.3 is selected from the group consisting of H and F; [0286] R.sup.4 is selected from the group consisting of F, Cl, Br and I; [0287] R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0288] R.sup.6 is selected from the group consisting of H; C.sub.1-5 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkynyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.3-6 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; phenyl optionally substituted with one or more, identical or different, substituents R.sup.9; and benzyl optionally substituted with one or more, identical or different, substituents R.sup.9; [0289] R.sup.7 is independently selected from the group consisting of deuterium, F and Cl; [0290] R.sup.8 is independently selected from the group consisting of deuterium, F, Cl, OC.sub.1-5 alkyl, SC.sub.1-5 alkyl, N(O)C.sub.1-5 alkyl and N(C.sub.1-5 alkyl)-C(O)C.sub.1-5 alkyl; and [0291] R.sup.9 is independently selected from the group consisting of deuterium, methoxy, nitro, cyano, Cl, Br, I, and F; [0292] or a pharmaceutically acceptable salt, hydrate, polymorph, tautomer, or solvate thereof for use in treating, ameliorating and/or preventing a neuromuscular disorder, and/or for use in reversing and/or ameliorating a neuromuscular blockade. [0293] 37. The compound for use according to item 36, wherein: [0294] R.sup.1 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; OC.sub.2-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0295] R.sup.2 is selected from the group consisting of C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 and C.sub.2-3 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.8 or when R.sup.1 is OC.sub.2-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 then R.sup.2 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 and C.sub.2-3 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.8; [0296] R.sup.3 is selected from the group consisting of H and F; [0297] R.sup.4 is selected from the group consisting of F, Cl, Br and I; [0298] R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0299] R.sup.6 is selected from the group consisting of H; C.sub.1-5 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkynyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.3-6 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; phenyl optionally substituted with one or more, identical or different, substituents R.sup.9; and benzyl optionally substituted with one or more, identical or different, substituents R.sup.9; [0300] R.sup.7 is independently selected from the group consisting of deuterium, F and Cl; [0301] R.sup.8 is independently selected from the group consisting of deuterium, F, Cl, OC.sub.1-5 alkyl, SC.sub.1-5 alkyl, N(O)C.sub.1-5 alkyl and N(C.sub.1-5 alkyl)-C(O)C.sub.1-5 alkyl; and [0302] R.sup.9 is independently selected from the group consisting of deuterium, methoxy, nitro, cyano, Cl, Br, I, and F; [0303] or a pharmaceutically acceptable salt, hydrate, polymorph, tautomer, or solvate thereof. [0304] 38. The compound for use according to item 37, with the proviso that when R.sup.1 is F or Me, R.sup.2 is Me, R.sup.3 is H, R.sup.4 is Cl or Br and R.sup.5 is OMe, then R.sup.6 is not H, Me or Et and with the proviso that when R.sup.1 is Cl or Br, R.sup.2 is Me, R.sup.3 is H, R.sup.4 is F and R.sup.5 is OMe, then R.sup.6 is not H, Me or Et and with the proviso that when R.sup.1 is F, R.sup.2 is Me, R.sup.3 is F, R.sup.4 is F and R.sup.5 is OMe, then R.sup.6 is not H. [0305] 39. The compound for use according to item 36, wherein: [0306] R.sup.1 is selected from the group consisting of F; Cl; Br; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; [0307] R.sup.2 is selected from the group consisting of C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; [0308] R.sup.3 is H; [0309] R.sup.4 is selected from the group consisting of F, Cl and Br; [0310] R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0311] R.sup.6 is H; [0312] R.sup.7 is F; and [0313] R.sup.8 is independently selected from the group consisting of deuterium, F, Cl, OC.sub.1-5 alkyl, SC.sub.1-5 alkyl, N(O)C.sub.1-5 alkyl and N(C.sub.1-5 alkyl)-C(O)C.sub.1-5 alkyl; [0314] or a pharmaceutically acceptable salt, hydrate, polymorph, tautomer, or solvate thereof. [0315] 40. The compound for use according to item 39, with the proviso that when R.sup.1 is F or Me, R.sup.2 is Me, R.sup.3 is H, R.sup.4 is Cl or Br and R.sup.5 is OMe, then R.sup.6 is not H and with the proviso that when R.sup.1 is Cl or Br, R.sup.2 is Me, R.sup.3 is H, R.sup.4 is F and R.sup.5 is OMe, then R.sup.6 is not H. [0316] 41. The compound for use according to any one of the preceding items, wherein R.sup.1 is selected from the group consisting of F; Cl; Br; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 and OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 [0317] 42. The compound for use according to any one of the preceding items, wherein R.sup.1 is selected from the group consisting of F, Cl, Br, C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 [0318] 43. The compound for use according to any one of the preceding items, wherein R.sup.1 is selected from the group consisting of MeO, Et, Me or Cl [0319] 44. The compound for use according to any one of the preceding items, wherein R.sup.1 is selected from the group consisting of Et, Me or Cl [0320] 45. The compound for use according to any one of the preceding items, wherein R.sup.1 is Me. [0321] 46. The compound for use according to any one of the preceding items, wherein if R.sup.1 is F; Cl; Br or I then R.sup.2 is not F; Cl; Br or I. [0322] 47. The compound for use according to any one of the preceding items, wherein R.sup.2 is C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 [0323] 48. The compound for use according to any one of the preceding items, wherein R.sup.2 is Cl or Me [0324] 49. The compound for use according to any one of the preceding items, wherein R.sup.2 is Me. [0325] 50. The compound for use according to any one of the preceding items, wherein R.sup.3 is H. [0326] 51. The compound for use according to any one of the preceding items, wherein R.sup.4 is selected from the group consisting of F, Cl and Br [0327] 52. The compound for use according to any one of the preceding items, wherein R.sup.4 is selected from the group consisting of F and Cl [0328] 53. The compound for use according to any one of the preceding items, wherein R.sup.4 is Cl. [0329] 54. The compound for use according to any one of the preceding items, wherein R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 [0330] 55. The compound for use according to any one of the preceding items, wherein R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7. [0331] 56. The compound for use according to any one of the preceding items, wherein R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 [0332] 57. The compound for use according to any one of the preceding items, wherein R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 [0333] 58. The compound for use according to any one of the preceding items, wherein R.sup.5 is selected from the group consisting of cyclopropoxy, OEt, OMe, OCH.sub.2F, OCHF.sub.2, OCF.sub.3, SMe, SCH.sub.2F, SCHF.sub.2 and SCF.sub.3 [0334] 59. The compound for use according to any one of the preceding items, wherein R.sup.5 is selected from the group consisting of OMe, OCH.sub.2F, OCHF.sub.2, OCF.sub.3 and SMe. [0335] 60. The compound for use according to any one of the preceding items, wherein R.sup.6 is H. [0336] 61. The compound for use according to any one of the preceding items, wherein R.sup.7 is F. [0337] 62. The compound for use according to any one of the preceding items, wherein R.sup.8 is SMe. [0338] 63. The compound for use according to any one of the preceding items, wherein the compound is selected from the group consisting of: [0339] 3-bromo-5-fluoro-2,6-dimethoxybenzoic acid; [0340] 3-fluoro-2,6-dimethoxy-5-methylbenzoic acid; [0341] 5-chloro-2-methyl-3-[(methylsulphanyl)methyl]benzoic acid; [0342] 3,5-dichloro-2,6-dimethoxybenzoic acid; [0343] 3-chloro-2-cyclopropoxy-5,6-dimethylbenzoic acid; [0344] 2,5-dichloro-6-methoxy-3-methylbenzoic acid; [0345] 3-chloro-2,6-dimethoxy-5-methylbenzoic acid; [0346] 3-chloro-5,6-dimethyl-2-(methylsulphanyl)benzoic acid; [0347] 3-chloro-5,6-dimethyl-2-(trifluoromethoxy)benzoic acid; [0348] 3-chloro-2-(difluoromethoxy)-5,6-dimethylbenzoic acid; [0349] 3-chloro-2-(fluoromethoxy)-5,6-dimethylbenzoic acid; [0350] 3-fluoro-2-methoxy-5,6-dimethylbenzoic acid; [0351] 3-chloro-2-ethoxy-5,6-dimethylbenzoic acid; [0352] 3-bromo-2-methoxy-5,6-dimethylbenzoic acid; and [0353] 3-chloro-2-methoxy-5,6-dimethylbenzoic acid. [0354] 64. The compound for use according to any one of the preceding items in the treatment of an indication selected from the group consisting of myasthenia gravis, autoimmune myasthenia gravis, congenital myasthenic syndrome, seronegative myasthenia gravis, muscle specific kinase myasthenia gravis (MuSK-MG), Lambert-Eaton Syndrome, critical illness myopathy, amyotrophic lateral sclerosis (ALS), spinal muscular atrophy (SMA), critical illness myopathy (CIM), Charcot-Marie Tooth disease, diabetic polyneuropathy, periodic paralysis, hypokalemic periodic paralysis, hyperkalemic periodic paralysis, myotubular myopathy, Duchenne muscular dystrophy, Guillain-Barre syndrome, poliomyelitis, post-polio syndrome, chronic fatigue syndrome, critical illness polyneuropathy, metabolic myopathy, Kennedy's disorder, multiple sclerosis and multifocal motor neuropathy. [0355] 65. The compound for use according to any one of the preceding items in the treatment of sarcopenia. [0356] 66. The compound for use according to any one of the preceding items in reversing and/or ameliorating a neuromuscular blockade wherein the neuromuscular blockade has been induced by a neuromuscular blocking agent. [0357] 67. The compound for use according to any one of the preceding items in the treatment of botulism poisoning, in the treatment of snake bites, in the treatment of nerve gas poisoning or prophylactically against nerve gas poisoning. [0358] 68. A method for treating and/or ameliorating a neuromuscular disorder, or for reversing and/or ameliorating a neuromuscular blockade comprising administering to a patient a therapeutically effective amount of a compound of Formula (I):
##STR00011## [0359] wherein: [0360] R.sup.1 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0361] R.sup.2 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 and C.sub.2-3 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.8; [0362] R.sup.3 is selected from the group consisting of H and F; [0363] R.sup.4 is selected from the group consisting of F, Cl, Br and I; [0364] R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0365] R.sup.6 is selected from the group consisting of H; C.sub.1-5 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkynyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.3-6 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; phenyl optionally substituted with one or more, identical or different, substituents R.sup.9; and benzyl optionally substituted with one or more, identical or different, substituents R.sup.9; [0366] R.sup.7 is independently selected from the group consisting of deuterium, F and Cl; [0367] R.sup.8 is independently selected from the group consisting of deuterium, F, Cl, OC.sub.1-5 alkyl, SC.sub.1-5 alkyl, N(O)C.sub.1-5 alkyl and N(C.sub.1-5 alkyl)-C(O)C.sub.1-5 alkyl; and [0368] R.sup.9 is independently selected from the group consisting of deuterium, methoxy, nitro, cyano, Cl, Br, I, and F; [0369] or a pharmaceutically acceptable salt, hydrate, polymorph, tautomer, or solvate thereof. [0370] 69. The method according to item 68, wherein: [0371] R.sup.1 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; OC.sub.2-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0372] R.sup.2 is selected from the group consisting of C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 and C.sub.2-3 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.8 or when R.sup.1 is OC.sub.2-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 then R.sup.2 is selected from the group consisting of F; Cl; Br; I; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 and C.sub.2-3 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.8; [0373] R.sup.3 is selected from the group consisting of H and F; [0374] R.sup.4 is selected from the group consisting of F, Cl, Br and I; [0375] R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0376] R.sup.6 is selected from the group consisting of H; C.sub.1-5 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkenyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.2-5 alkynyl optionally substituted with one or more, identical or different, substituents R.sup.7; C.sub.3-6 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; phenyl optionally substituted with one or more, identical or different, substituents R.sup.9; and benzyl optionally substituted with one or more, identical or different, substituents R.sup.9; [0377] R.sup.7 is independently selected from the group consisting of deuterium, F and Cl; [0378] R.sup.8 is independently selected from the group consisting of deuterium, F, Cl, OC.sub.1-5 alkyl, SC.sub.1-5 alkyl, N(O)C.sub.1-5 alkyl and N(C.sub.1-5 alkyl)-C(O)C.sub.1-5 alkyl; and [0379] R.sup.9 is independently selected from the group consisting of deuterium, methoxy, nitro, cyano, Cl, Br, I, and F; [0380] or a pharmaceutically acceptable salt, hydrate, polymorph, tautomer, or solvate thereof. [0381] 70. The method according to any one of items 68 or 69, wherein: [0382] R.sup.1 is selected from the group consisting of F; Cl; Br; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; [0383] R.sup.2 is selected from the group consisting of C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8; [0384] R.sup.3 is H; [0385] R.sup.4 is selected from the group consisting of F, Cl and Br; [0386] R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7; SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; [0387] R.sup.6 is H; [0388] R.sup.7 is F; and [0389] R.sup.8 is independently selected from the group consisting of deuterium, F, Cl, OC.sub.1-5 alkyl, SC.sub.1-5 alkyl, N(O)C.sub.1-5 alkyl and N(C.sub.1-5 alkyl)-C(O)C.sub.1-5 alkyl; [0390] or a pharmaceutically acceptable salt, hydrate, polymorph, tautomer, or solvate thereof. [0391] 71. The method according to any one of items 68, 69 or 70, wherein R.sup.1 is selected from the group consisting of F; Cl; Br; C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 and OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 [0392] 72. The method according to any one of items 68, 69 or 70, wherein R.sup.1 is selected from the group consisting of F, Cl, Br, C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 [0393] 73. The method according to any one of items 68, 69 or 70, wherein R.sup.1 is selected from the group consisting of MeO, Et, Me or Cl [0394] 74. The method according to any one of items 68, 69 or 70, wherein R.sup.1 is selected from the group consisting of Et, Me or Cl [0395] 75. The method according to any one of items 68, 69 or 70, wherein R.sup.1 is Me. [0396] 76. The method according to any one of items 68, 69, 70, 71, 72, 73, 74 or 75, wherein if R.sup.1 is F; Cl; Br or I then R.sup.2 is not F; Cl; Br or I. [0397] 77. The method according to any one of items 68, 69, 70, 71, 72, 73, 74, 75 or 76, wherein R.sup.2 is C.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.8 [0398] 78. The method according to any one of items 68, 69, 70, 71, 72, 73, 74, 75 or 76, wherein R.sup.2 is Cl or Me [0399] 79. The method according to any one of items 68, 69, 70, 71, 72, 73, 74, 75 or 76, wherein R.sup.2 is Me. [0400] 80. The method according to any one of items 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78 or 79, wherein R.sup.3 is H. [0401] 81. The method according to any one of items 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79 or 80, wherein R.sup.4 is selected from the group consisting of F, Cl and Br [0402] 82. The method according to any one of items 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79 or 80, wherein R.sup.4 is selected from the group consisting of F and Cl [0403] 83. The method according to any one of items 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79 or 80, wherein R.sup.4 is Cl. [0404] 84. The method according to any one of items 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82 or 83, wherein R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7; OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 [0405] 85. The method according to any one of items 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82 or 83, wherein R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and OC.sub.3-5 cycloalkyl optionally substituted with one or more, identical or different, substituents R.sup.7. [0406] 86. The method according to any one of items 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82 or 83, wherein R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 and SC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 [0407] 87. The method according to any one of items 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82 or 83, wherein R.sup.5 is selected from the group consisting of OC.sub.1-3 alkyl optionally substituted with one or more, identical or different, substituents R.sup.7 [0408] 88. The method according to any one of items 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82 or 83, wherein R.sup.5 is selected from the group consisting of cyclopropoxy,-OEt, OMe, OCH.sub.2F, OCHF.sub.2, OCF.sub.3, SMe, SCH.sub.2F, SCHF.sub.2 and SCF.sub.3 [0409] 89. The method according to any one of items 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82 or 83, wherein R.sup.5 is selected from the group consisting of OMe, OCH.sub.2F, OCHF.sub.2, OCF.sub.3 and SMe. [0410] 90. The method according to any one of items 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88 or 89, wherein R.sup.6 is H. [0411] 91. The method according to any one of items 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89 or 90, wherein R.sup.7 is F. [0412] 92. The method according to any one of items 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90 or 91, wherein R.sup.8 is SMe. [0413] 93. The method according to any one of items 68, wherein the compound is selected from the group consisting of: [0414] 3-bromo-5-fluoro-2,6-dimethoxybenzoic acid; [0415] 3-fluoro-2,6-dimethoxy-5-methylbenzoic acid; [0416] 5-chloro-2-methyl-3-[(methylsulphanyl)methyl]benzoic acid; [0417] 3,5-dichloro-2,6-dimethoxybenzoic acid; [0418] 3-chloro-2-cyclopropoxy-5,6-dimethylbenzoic acid; [0419] 2,5-dichloro-6-methoxy-3-methylbenzoic acid; [0420] 3-chloro-2,6-dimethoxy-5-methylbenzoic acid; [0421] 3-chloro-5,6-dimethyl-2-(methylsulphanyl)benzoic acid; [0422] 3-chloro-5,6-dimethyl-2-(trifluoromethoxy)benzoic acid; [0423] 3-chloro-2-(difluoromethoxy)-5,6-dimethylbenzoic acid; [0424] 3-chloro-2-(fluoromethoxy)-5,6-dimethylbenzoic acid; [0425] 3-fluoro-2-methoxy-5,6-dimethylbenzoic acid; [0426] 3-chloro-2-ethoxy-5,6-dimethylbenzoic acid; [0427] 3-bromo-2-methoxy-5,6-dimethylbenzoic acid; and [0428] 3-chloro-2-methoxy-5,6-dimethylbenzoic acid. [0429] 94. The method according to any one of items 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92 or 93, wherein the compound is an inhibitor of the CIC-1 ion channel. [0430] 95. The method of item 68, wherein said neuromuscular disorder is one selected from the group consisting of myasthenia gravis, autoimmune myasthenia gravis, congenital myasthenic syndrome, seronegative myasthenia gravis, muscle specific kinase myasthenia gravis (MuSK-MG), Lambert-Eaton Syndrome, critical illness myopathy, amyotrophic lateral sclerosis (ALS), spinal muscular atrophy (SMA), critical illness myopathy (CIM), Charcot-Marie Tooth disease, diabetic polyneuropathy, periodic paralysis, hypokalemic periodic paralysis, hyperkalemic periodic paralysis, myotubular myopathy, Duchenne muscular dystrophy, Guillain-Barre syndrome, poliomyelitis, post-polio syndrome, chronic fatigue syndrome, critical illness polyneuropathy, metabolic myopathy, Kennedy's disorder, multiple sclerosis and multifocal motor neuropathy. [0431] 96. The method of item 68, wherein said neuromuscular disorder is sarcopenia. [0432] 97. The method of item 68, wherein said method is for reversing and/or ameliorating a neuromuscular blockade induced by a neuromuscular blocking agent. [0433] 98. The method of item 68, wherein said neuromuscular disorder is due to botulism poisoning, snake bites, nerve gas poisoning or prophylactically against nerve gas poisoning. [0434] 99. The compound for use according to any one of the preceding items, wherein said compound is comprised in a composition. [0435] 100. The compound for use according to any one of the preceding items, wherein the composition is a pharmaceutical composition. [0436] 101. The compound for use according to any one of the preceding items, wherein the composition further comprises a pharmaceutically acceptable carrier. [0437] 102. The compound for use according to any one of items, wherein the composition further comprises at least one further active agent. [0438] 103. The compound for use according to any one of the preceding items, wherein said further active agent is suitable for treating, preventing or ameliorating said neuromuscular disorder. [0439] 104. The compound for use according to any one of the preceding items, wherein said further active agent is an acetylcholine esterase inhibitor. [0440] 105. The compound for use according to any one of the preceding items, wherein said acetylcholine esterase inhibitor is selected from the group consisting of delta-9-tetrahydrocannabinol, carbamates, physostigmine, neostigmine, pyridostigmine, ambenonium, demecarium, rivastigmine, phenanthrene derivatives, galantamine, piperidines, donepezil, tacrine, edrophonium, huperzine, ladostigil, ungeremine and lactucopicrin. [0441] 106. The compound for use according to any one of the preceding items, wherein said acetylcholine esterase inhibitor is neostigmine or pyridostigmine. [0442] 107. The compound for use according to any one of the preceding items, wherein said further active agent is sugammadex. [0443] 108. The compound for use according to any one of the preceding items, wherein said further active agent is 3,4-diaminopyridine. [0444] 109. A method of treating, preventing and/or ameliorating a neuromuscular disorder, said method comprising administering a therapeutically effective amount of the compound as defined in any one of the preceding items to a person in need thereof. [0445] 110. Use of a compound as defined in any one of the preceding items, for the manufacture of a medicament for the treatment, prevention and/or amelioration of a neuromuscular disorder, and/or for reversing and/or ameliorating of a neuromuscular blockade. [0446] 111. A method of reversing and/or ameliorating a neuromuscular blockade, said method comprising administering a therapeutically effective amount of the compound as defined in any one of the preceding items to a person in need thereof. [0447] 112. A method for recovery of neuromuscular transmission, said method comprising administering a therapeutically effective amount of the compound as defined in any one of the preceding items to a person in need thereof. [0448] 113. A method for recovering neuromuscular transmission, the method comprising administering a compound as defined in any one of the preceding items to an individual in need thereof.
EXAMPLES
General Synthetic Strategies
[0449] General methods for the synthesis of carboxylic acids and substitution on aromatic rings are featured in literature sources such as: March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 8th Edition, Michael B. Smith, Ed.; ISBN: 978-1-119-37179-3; John Wiley, 2020.
Materials and Methods
NMR Spectra
[0450] .sup.1H-NMR and .sup.19F-NMR spectra were recorded on a Bruker AM-300 spectrometer and were calibrated using residual nondeuterated solvent as internal reference. Spectra were processed using Spinworks version 4.0 (developed by Dr. Kirk Marat, Department of Chemistry, University of Manitoba).
LC/MS System
[0451] Samples were analysed by direct inject on a Waters Micromass Quattro Ultima with a Waters 2695 HPLC. Mass spectra were recorded in ESI scan mode (negative/positive). Mass spectra were processed using Water MassLynx Mass Spectrometry Software.
HPLC Method
[0452] The product was analysed by a Waters 2695 HPLC consisting of a Waters 996 photodiode array detector (Waters, Milford, MA, USA) and a Kromasil Eternity C18, 5 m, 4.6150 mm column. Flow rate: 1 mL/minute, run time 20 minutes. Spectra were processed using Waters Empower Software. The chromatographic system was operated using gradient elution with HPLC grade solvents methanol (solvent A), 0.1% formic acid in water (solvent B), and acetonitrile (solvent C) at ambient temperature and UV detection at either 280 nm or 254 nm. HPLC gradients were either 0% of A to 100% of A over 15 minutes or 10% of C to 100% of C over 15 minutes or gradient 10% of C to 100% of C over 7 minutes.
Example 1: Synthesis of 3-fluoro-2,6-dimethoxy-5-methylbenzoic acid (A-1)
##STR00012##
Step 1: 3-Fluoro-2,6-dianisic acid
[0453] To a solution of 1-fluoro-2,4-dimethoxybenzene (1.0 g, 6.40 mmol) in THF (10 mL) at 78 C. was added n-BuLi (2.5 M in hexane) (2.82 mL, 7.04 mmol). The reaction mixture was stirred at that temperature for 1 h, and CO.sub.2 (g) was bubbled through it for 10 min. and stirred for 1 h at 78 C. The temperature of the mixture allowed to reach 0 C. and quenched 1M NaOH until the pH reached 11. The reaction mixture was washed with CH.sub.2Cl.sub.2 (210 mL) to remove impurities, and the aqueous layer was acidified with 1N HCl to a pH value of 1. Extraction with EtOAc (50 mL), drying (Na.sub.2SO.sub.4) filtration and evaporation in vacuo provided 3-fluoro-2,6-dianisic acid (1.0 g) which was utilised directly in the next step without further purification.
Step 2: Methyl 3-fluoro-2,6-dianisate
[0454] Potassium carbonate (2.07 g, 14.99 mmol) was introduced to a solution of 3-fluoro-2,6-dianisic acid (1.0 g, 4.99 mmol) in dimethylformamide (15 mL) and the mixture was stirred at ambient temperature for 20 min. Iodomethane (0.467 mL, 7.49 mmol) was added and the reaction mixture stirred for 18 h at ambient temperature. The suspension was quenched with water, extracted with EtOAc (50 mL) and the extract treated with brine (220 mL). The organic layer was separated, dried over Na.sub.2SO.sub.4, and concentrated. The crude product was purified by column chromatography on silica gel eluting with (0-30%) EtOAc/hexane to afford methyl 3-fluoro-2,6-dianisate (0.354 g: 33% yield).
[0455] .sup.1H NMR (300 MHZ, CDCl.sub.3) 7.25 (dd, 1H), 6.74 (dd, 1H), 4.14 (d, 3H), 4.11 (s, 3H), 3.97 (s, 3H).
[0456] .sup.19F NMR (300 MHZ, CDCl3) 139.78 ppm.
Step 3: Methyl 3-bromo-5-fluoro-2,6-dianisate
[0457] To a solution of methyl 3-fluoro-2,6-dianisate (0.35 g, 1.64 mmol) in CH.sub.3CN (5 mL) was added N-bromosuccinimide (0.32 g, 1.79 mmol) and the reaction mixture was stirred at ambient temperature for 4 days. The reaction mixture was concentrated, extracted with EtOAc (30 mL), washed with water (210 mL), and brine (15 mL). The organic layer was dried (Na.sub.2SO.sub.4), concentrated in vacuo and the residue was purified by column chromatography on silica gel utilising (0-30%) EtOAc/hexane to provide methyl 3-bromo-5-fluoro-2,6-dianisate (0.348 g: 73% yield).
[0458] .sup.1H NMR (300 MHZ, CDCl.sub.3) 7.24 (dd, 1H), 3.86-3.81 (br, 6H), 3.74 (s, 3H).
[0459] .sup.19F NMR (300 MHZ, CDCl3) 132.05 ppm.
Step 4: Methyl 5-fluoro-2,4-dimethoxy-3-toluate
[0460] A solution of methyl 3-bromo-5-fluoro-2,6-dianisate (100 mg, 0.34 mmol) in 1,4-dioxane (2 mL) in a microwave vial was degassed with argon for 20 min. Methylboronic acid (24.6 mg, 0.4 mmol), Pd(dppf)Cl.sub.2 (12.5 mg, 0.017 mmol) and K.sub.2CO.sub.3 (70.8 mg, 0.51 mmol) were added then the reaction mixture was subjected to microwave radiation at 130 C. for 1 h. The mixture was extracted with EtOAc (30 mL) which was washed with water (210 mL) and brine (315 mL). The organic layer was dried (Na.sub.2SO.sub.4) concentrated, and the crude product was purified by column chromatography on silica gel eluting with (0-20% EtOAc/hexane) to afford methyl 5-fluoro-2,4-dimethoxy-3-toluate (48.0 mg: 61% yield).
[0461] .sup.1H NMR (300 MHZ, CDCl.sub.3) 7.24 (dd, 1H), 4.22 (s, 3H), 4.20 (d, 3H), 4.04 (s, 3H), 2.51 (s, 3H).
[0462] .sup.19F NMR (300 MHz, CDCl3) 135.25 ppm.
Step 5: 3-fluoro-2,6-dimethoxy-5-methylbenzoic acid
[0463] To a solution of methyl 5-fluoro-2,4-dimethoxy-3-toluate (48.0 mg, 0.21 mmol) in CH.sub.3OH (2 mL) in a microwave vial was added NaOH (9.26 mg, 0.23 mmol) pre-dissolved in water (1 mL). The reaction was subjected to microwave at 130 C. for 30 min, concentrated, redissolved in water (2 mL), and washed with CH.sub.2Cl.sub.2 (210 mL) to remove organic impurities. The aqueous layer was acidified with 1N HCl to PH 1, extracted with EtOAc (20 mL), dried (Na.sub.2SO.sub.4), filtered and concentrated to obtain 3-fluoro-2,6-dimethoxy-5-methylbenzoic acid (31.0 mg: 68% yield).
[0464] .sup.1H NMR (300 MHZ, CD.sub.3OD) 7.06 (dd, 1H), 3.90 (d, 3H), 3.78 (s, 3H), 2.25 (s, 3H). [0465] .sup.19F NMR (300 MHZ, CDCl3) 137.37 ppm.
[0466] HPLC: Retention time 6.143 min, purity >98% at 280 nm.
Example 2: Synthesis of 3-bromo-5-fluoro-2,6-dimethoxybenzoic acid (A-2)
##STR00013##
Step 1: 3-Bromo-5-fluoro-2,6-dimethoxybenzoic acid
[0467] A solution of NaOH (15.07 mg, 0.38 mmol) in water (1 ml) was added to a solution of methyl 3-bromo-5-fluoro-2,6-dimethoxybenzoate (100.0 mg, 0.34 mmol) in CH.sub.3OH (2 mL) and the reaction mixture was subjected to microwave conditions at 130 C. for 30 min. The mixture was evaporated to a residue, redissolved in water (2 ml), and washed with CH.sub.2Cl.sub.2 (210 mL). The aqueous layer was acidified with 1N HCl to PH 1 and extracted with EtOAc (20 mL). The solution was dried (Na.sub.2SO.sub.4), filtered and evaporated to obtain the title compound (76.0 mg, 80% yield).
[0468] 1H NMR (300 MHZ, CD3OD) 7.52 (d, 1H), 3.95 (d, 3H), 3.87 (s, 3H).
[0469] 19F NMR (300 MHZ, CD3OD) 133.95 ppm.
[0470] HPLC: Retention time 6.519 min, purity >98% at 280 nm.
Example 3: 3,5-Dichloro-2,6-dimethoxybenzoic acid (A-3)
[0471] Compound is commercially available from e.g. Merck (PH006804).
Example 4: Synthesis of 3-chloro-5,6-dimethyl-2-(trifluoromethoxy)benzoic acid (A-4)
##STR00014##
Step 1: 2-Bromo-6-chloro-3,4-xylenol
[0472] To a solution of 2-chloro-4,5-xylenol (1.0 g, 6.41 mmol) in CH.sub.3CN (25 mL) N-bromosuccinimide (1.14 g, 6.41 mmol) was introduced and the reaction mixture was stirred at ambient temperature for 18 h. It was then concentrated in vacuo, extracted with EtOAc (50 mL) and the solution washed with a saturated solution of NaHCO.sub.3 (20 mL) and water (230 mL). The organic layer was dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with (0-20%) EtOAc/hexane to afford 2-bromo-6-chloro-3,4-xylenol (1.45 g: 97% yield).
[0473] .sup.1H NMR (300 MHZ, CDCl.sub.3) 7.13 (s, 1H), 6.14 (s, 1H), 2.40 (s, 3H), 2.33 (s, 3H).
Step 2: O-2-Bromo-6-chloro-3,4-dimethylphenyl S-methyl carbonodithioate
[0474] To a solution of 2-bromo-6-chloro-3,4-xylenol (0.50 g, 2.14 mmol) and triethylamine (0.327 mL, 2.35 mmol) in CH.sub.3CN (10 mL) at 0 C., 3-methyl-1-(methylthiocarbonothioyl)-1H-imidazol-3-ium (0.37 g, 2.14 mmol) was added. The reaction was stirred at ambient temperature for 18 h. The reaction mixture was concentrated and extracted with EtOAc (50 mL), washed with saturated solution of NaHCO.sub.3 (20 mL), and water (230 mL). The organic layer was dried over Na.sub.2SO.sub.4, concentrated in vacuo and the residue was purified by column chromatography on silica gel eluting with (0-20%) EtOAc/hexane to provide O-2-bromo-6-chloro-3,4-dimethylphenyl-S-methyl carbonodithioate (0.61 g: 87% yield).
[0475] 1H NMR (300 MHZ, CDCl.sub.3) 7.27 (s, 1H), 2.78 (s, 3H), 2.43 (s, 3H), 2.39 (s, 3H).
Step 3: 3-Bromo-1-chloro-2-trifluoromethoxy-4,5-xylene
[0476] A mixture of O-2-bromo-6-chloro-3,4-dimethylphenyl-S-methyl-carbonodithioate (0.6 g, 1.85 mmol), XtalFluor-E (1.27 g, 5.56 mmol), and N-fluorobenzenesulphonimide (1.75 g, 5.56 mmol) in 1,2-dichloroethane (10 mL) was heated at 80 C. for 48 h. The reaction mixture was treated with a saturated solution of NaHCO.sub.3 (10 mL), filtered through a celite pad and extracted with EtOAc (230 mL). The organic layer was washed with water (220 mL), dried over Na.sub.2SO.sub.4, concentrated in vacuo and the product purified by column chromatography on silica gel, eluting with (0-20%) EtOAc/hexane to provide 3-bromo-1-chloro-2-trifluoromethoxy-4,5-xylene (0.10 g: 17.9% yield).
[0477] .sup.1H NMR (300 MHz, CDCl.sub.3) 7.28 (s, 1H), 2.44 (s, 3H), 2.38 (s, 3H).
[0478] .sup.19F NMR (300 MHZ, CDCl3) 55.206 ppm.
Step 4: 3-Chloro-5,6-dimethyl-2-(trifluoromethoxy)benzoic acid
[0479] n-BuLi (2.5 M in hexane) (0.073 mL, 0.182 mmol) was added to a solution of 3-bromo-1-chloro-2-trifluoromethoxy-4,5-xylene (50 mg, 0.165 mmol) in THF (4 mL) at 78 C. The reaction mixture was stirred at that temperature for 1 h and then CO.sub.2 gas was bubbled through for 10 min and it was stirred for a further 1 h at 78 C. The reaction temperature was elevated to 0 C. and the pH was adjusted with 1M NaOH to 11. The mixture was extracted with CH2Cl2 (210 mL) to remove organic impurities; the water layer was acidified with 1N HCl to PH 1 and then extracted with EtOAc (20 mL). The solution was dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude product was purified by semi-prep HPLC utilising 30 to 100% acetonitrile/0.1% formic acid in water to afford 3-chloro-5,6-dimethyl-2-(trifluoromethoxy)benzoic acid (6.5 mg: 14.6% yield).
[0480] .sup.1H NMR (300 MHZ, CD.sub.3OD) 7.24 (s, 1H), 2.19 (s, 3H), 2.17 (s, 3H).
[0481] .sup.19F NMR (300 MHZ, CD.sub.3OD) 57.560 ppm.
[0482] HPLC: Retention time 7.305 min, purity >98% at 280 nm.
Example 5: Synthesis of 3-chloro-2-cyclopropoxy-5,6-dimethylbenzoic acid (A-5)
##STR00015##
Step 1: 3-Bromo-1-chloro-2-cyclopropoxy-4,5-dimethylbenzene
[0483] To a solution of 2-bromo-6-chloro-3,4-dimethylphenol (0.4 g, 1.7 mmol) in dimethyl acetamide (3 mL), cesium carbonate (1.66 g, 5.1 mmol, 3.0 eq.) was introduced, followed by bromocyclopropane (0.62 g, 5.1 mmol, 3.0 eq.). The reaction mixture was heated at 150 C. for 72 h, cooled, extracted with EtOAc (220 mL), then the combined extracts were washed with water (210 mL) and brine (10 mL). The organic layer was dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The crude product was purified by column chromatography on silica gel eluting with (0-5%) EtOAc/hexane to provide 3-bromo-1-chloro-2-cyclopropoxy-4,5-dimethylbenzene (0.38 g, 81% yield).
[0484] .sup.1H NMR (300 MHz, CDCl.sub.3) 7.13 (s, 1H), 4.44-4.36 (m, 1H), 2.35 (s, 3H), 2.27 (s, 3H), 1.06-0.97 (m, 2H), 0.60-0.50 (m, 2H).
Step 2: 3-Chloro-2-cyclopropoxy-5,6-dimethylbenzoic acid
[0485] To a solution of 3-bromo-1-chloro-2-cyclopropoxy-4,5-dimethylbenzene (0.2 g, 0.73 mmol) in THF (8 mL) at 78 C., n-BuLi (2.5 M in hexane) (0.32 mL, 0.8 mmol, 1.1 eq.) was added. The reaction mixture was stirred at that temperature for 30 min. After a further 30 min. CO.sub.2 (g) was bubbled through the reaction mixture for 10 min. which was stirred for 1 h at 78 C. and slowly brought to 0 C. prior to quenching with 1M NaOH to pH 11. Washing with EtOAc (10 mL) was followed by separation of the water layer which was acidified with 1N HCl to PH 1. Extraction with EtOAc (210 mL) took place, and the combined extracts were washed with water (10 mL) and brine (10 mL). The organic layer was dried (Na.sub.2SO.sub.4), filtered and concentrated in vacuo to obtain 3-chloro-2-cyclopropoxy-5,6-dimethylbenzoic acid (0.083 g, 47% yield) as a white solid.
[0486] .sup.1H NMR (300 MHz, CDCl.sub.3) 10.55-9.50 (Br s, 1H), 7.25 (s, 1H), 4.41-4.33 (m, 1H), 2.26 (s, 3H), 2.25 (s, 3H), 0.98-0.90 (m, 2H), 0.62-0.53 (m, 2H).
[0487] ES-MS: m/z 239.3 (M1).
[0488] HPLC: Retention time 7.262 min, purity >98% at 280 nm.
Example 6: Synthesis of 2,5-dichloro-6-methoxy-3-methylbenzoic acid (A-6)
##STR00016##
Step 1: 3-Bromo-2-chloro-5-nitro-4-cresol
[0489] To a solution of 2-chloro-5-nitro-4-cresol (1.0 g, 5.35 mmol) in CH.sub.3CN (10 mL) was added N-bromosuccinimide (952 mg, 5.35 mmol, 1.0 eq.). The reaction mixture was stirred at ambient temperature for 16 h. The reaction mixture was concentrated and extracted with EtOAc (250 mL) then the combined extracts were washed with water (350 mL) and brine (15 mL) before drying (Na.sub.2SO.sub.4). The crude product was obtained by evaporation in vacuo and purified by column chromatography on silica gel eluting with (0-5%) EtOAc/hexane to provide 3-bromo-2-chloro-5-nitro-4-cresol (1.50 g, 96%).
[0490] .sup.1H NMR (300 MHZ, CDCl.sub.3) 8.03 (br s, 1H), 8.02 (s, 1H), 2.42 (s, 3H) ppm.
Step 2: 3-Bromo-2-chloro-4-methoxy-5-nitrotoluene
[0491] To a solution of 3-bromo-2-chloro-5-nitro-4-cresol (1.50 g, 5.64 mmol) in DMF (8 mL), K.sub.2CO.sub.3 (1.56 g, 11.28 mmol, 2 eq.) was added, followed by iodomethane (1.43 mL, 22.56 mmol, 4 eq.). The reaction mixture was stirred at ambient temperature for 18 h and the solvent was removed in vacuo and 2N HCl solution (20 mL) was added. Extraction with CH.sub.2Cl.sub.2 (330 mL) and drying (Na.sub.2SO.sub.4) provided the crude 3-bromo-2-chloro-4-methoxy-5-nitrotoluene after evaporation in vacuo as an oil (1.53 g, 96%).
[0492] .sup.1H NMR (300 MHZ, CDCl.sub.3) 7.69 (s, 1H), 3.97 (s, 3H), 2.44 (s, 3H).
Step 3: 3-Bromo-4-chloro-2-methoxytolylamine
[0493] To a solution of 3-bromo-2-chloro-4-methoxy-5-nitrotoluene (2.0 g, 7.14 mmol, 1.0 eq.) in MeOH (100 mL) and water (100 mL), was added ammonium chloride solution (4.54 g, 85.68 mmol, 12 eq.), followed by Fe powder (2.4 g, 42.86 mmol). The reaction mixture was stirred at ambient temperature for 16 h and filtered through a plug of celite. The solvent was removed in vacuo, saturated NaHCO.sub.3 solution (20 mL) was added and the mixture was extracted with CH.sub.2Cl.sub.2 (3100 mL). The combined extracts were dried (Na.sub.2SO.sub.4) and concentrated to provide 3-bromo-4-chloro-2-methoxytolylamine as an oil 1.66 g (93% yield).
[0494] .sup.1H NMR (300 MHZ, CDCl.sub.3) 6.59 (s, 1H), 3.95 (br, 2H), 3.86 (s, 3H), 2.30 (s, 3H).
Step 4: 3-Bromo-2,5-dichloro-4-methoxytoluene
[0495] CuCl.sub.2 (1.66 g, 7.03 mmol) in CH.sub.3CN was cooled to 0 C. and t-BuONO (1.253 mL, 10.55 mmol, 1.5 eq.) was added. After 2 min, 3-bromo-4-chloro-2-methoxytolylamine (1.66, 7.034 mL) in CH.sub.3CN was added slowly and the reaction mixture was stirred for 30 min at 0 C. and then for 3 h. at ambient temperature. 2N HCl solution was added, the mixture was stirred for 30 min and the product extracted with EtOAc (3100 mL) and the combined extracts were dried (Na.sub.2SO.sub.4) and concentrated in vacuo. Purification by silica gel chromatography eluting with (0-5%) EtOAc/hexane to afford 3-bromo-2,5-dichloro-4-methoxytoluene as a yellow solid (1.63 g, 86% yield).
[0496] .sup.1H NMR (300 MHZ, CDCl.sub.3) 7.26 (s, 1H), 3.90 (s, 3H), 2.40 (s, 3H).
Step 5: 2,5-Dichloro-6-methoxy-3-methylbenzoic acid
[0497] To a solution of 3-bromo-2,5-dichloro-4-methoxytoluene (0.5 g, 1.865 mmol) in THF (10 mL) at 78 C. was added n-BuLi (2.5 M in hexane) (0.874 mL, 1.96 mmol, 1.05 eq.). The reaction mixture was stirred at that temperature for 50 min and CO.sub.2 (g) was bubbled through it for 10 min. It was stirred for 1 h at 78 C., and slowly allowed to reach 0 C. before being quenched with 1M NaOH to reach PH 11. After washing with EtOAc (10 mL) the water layer was separated and acidified with 1N HCl to PH 1. Extraction was performed with EtOAc (350 mL) and the combined organic extracts were dried (Na.sub.2SO.sub.4) filtered then concentrated in vacuo to afford 2,5-dichloro-6-methoxy-3-methylbenzoic acid (0.3 g, 68%) as a white solid.
[0498] 1H NMR (300 MHZ, CD3OD) 7.25 (s, 1H), 3.92 (s, 3H), 2.33 (s, 3H).
[0499] ES-MS: m/z 233.8 (M1).
[0500] HPLC: Retention time 10.082 min, purity 96% at 280 nm.
Example 7: Synthesis of 3-chloro-2,6-dimethoxy-5-methylbenzoic acid (A-7)
##STR00017##
Step 1: 2,6-Dimethoxy-3-methylbenzoic acid
[0501] To a solution of 2,4-dimethoxy-1-methylbenzene (2.00 g, 13.1 mmol) in THF (20 mL) at 0 C. was added n-BuLi (9.1 mL, 14.5 mmol, [1.6 M]), the reaction mixture was stirred at ambient temperature for 1 h. The reaction mixture was cooled to 0 C. and CO.sub.2 gas was bubbled through it over a period of 45 min. The reaction mixture was quenched with water (25 mL) and washed with EtOAc (225 mL). The aqueous layer was acidified to pH 2 using 1.0 N HCl and extracted with EtOAc (225 mL). The combined organic extracts were dried (Na.sub.2SO.sub.4) and concentrated under reduced pressure to provide 2,6-dimethoxy-3-methylbenzoic acid (1.72 g, 67%).
[0502] .sup.1H NMR (300 MHZ, CDCl.sub.3) 7.27 (d, J=8.4 Hz, 1H), 6.72 (d, J=8.4 Hz, 1H), 3.93 (s, 3H), 3.92 (s, 3H), 2.32 (s, 3H).
[0503] ES-MS: 195.6 [MH]
Step 2: 3-Chloro-2,6-dimethoxy-5-methylbenzoic acid
[0504] To a solution of 2,6-dimethoxy-3-methylbenzoic acid (1.72 g, 8.77 mmol) in CHCl.sub.3 (20 mL) at 0 C. ethanol (0.1 mL) was introduced, followed by sulphuryl chloride (1.41 mL, 17.5 mmol). The reaction mixture was stirred at ambient temperature for 24 h and the solvent was removed under reduced pressure. The residue was diluted with EtOAc (50 mL) and extracted with 1.0 N NaOH (250 mL). The combined aqueous extracts were acidified to PH 2 with 1.0 N HCl and extracted with EtOAc (250 mL). The combined organic extracts were dried (Na.sub.2SO.sub.4) and concentrated under reduced pressure. The crude product was subjected to column chromatography on silica gel eluting with 20% EtOAc/hexanes) to afford 3-chloro-2,6-dimethoxy-5-methylbenzoic acid (0.40 g, 20%).
[0505] .sup.1H NMR (300 MHZ, CDCl.sub.3) 7.28 (s, 1H), 3.93 (s, 3H), 3.84 (s, 3H), 2.25 (s, 3H).
[0506] ES-MS: 231.1 [M+H]
[0507] HPLC Retention Time: 9.60; purity >98% at 280 nm
Example 8: Synthesis of 3-chloro-5,6-dimethyl-2-(methylsulphanyl)benzoic acid (A-8)
##STR00018##
Step 1: 2-Bromo-6-chloro-3,4-dimethylaniline
[0508] To a solution of 2-chloro-4,5-dimethylaniline (2.0 g, 12.9 mmol) in DMF (20 mL), N-bromosuccinimide (2.29 g, 12.9 mmol, 1.0 eq.) was added. The reaction mixture was stirred at ambient temperature for 16 h, concentrated in vacuo then water was added (20 mL). The mixture was extracted with EtOAc (240 mL) then the combined extracts were washed with water (320 mL) and brine (15 mL). The combined organic extracts were dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The crude product was purified by column chromatography on silica gel eluting with (0-20%) EtOAc/hexane to provide 2-bromo-6-chloro-3,4-dimethylaniline (2.6 g, 86% yield).
[0509] .sup.1H NMR (300 MHZ, CD.sub.3OD) 7.04 (s, 1H), 2.32 (s, 3H), 2.22 (s, 3H).
Step 2: (2-Bromo-6-chloro-3,4-dimethylphenyl)(methyl)sulphane
[0510] 2-Bromo-6-chloro-3,4-dimethylaniline (0.8 g, 3.43 mmol, 1.0 eq.) in chloroform (6 mL) and tert-butyl nitrite (0.48 g, 4.64 mmol, 1.35 eq.) was simultaneously added into a solution of dimethyl disulphide (0.71 g, 7.55 mmol, 2.2 eq.) in chloroform (2 mL). The reaction mixture was stirred at ambient temperature for 30 min, extracted with chloroform (220 mL) and the combined extracts washed with water (10 mL), 1N HCl (10 mL) and brine (10 mL). The organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude product was purified by column chromatography on silica gel using (0-10% EtOAc/hexane) to obtain (2-bromo-6-chloro-3,4-dimethylphenyl)(methyl)sulphane (0.52 g, 62% yield).
[0511] .sup.1H NMR (300 MHZ, CDCl.sub.3) 7.25 (s, 1H), 2.41 (s, 3H), 2.40 (s, 3H), 2.32 (s, 3H).
Step 3: 3-Chloro-5,6-dimethyl-2-(methylsulphanyl)benzoic acid
[0512] To a solution of (2-bromo-6-chloro-3,4-dimethylphenyl)(methyl)sulphane (0.075 g, 0.307 mmol) in diethyl ether (3 mL) at 78 C., n-BuLi (2.5 M in hexane) (0.135 mL, 0.338 mmol, 1.1 eq.) was added. The mixture was stirred at that temperature for 1 h and after 1 h CO.sub.2 (g) was bubbled through the reaction mixture for 10 min and stirred for 1 h at 78 C., slowly brought to 0 C. and quenched with 1M NaOH to PH 11. It was washed with EtOAc (5 mL) to remove non-acid impurities and water layer was separated then acidified with 1N HCl to PH 1 before extraction with EtOAc (210 mL) The combined extracts were washed with water (5 mL), brine (5 mL) and dried (Na.sub.2SO.sub.4), filtered and concentrated in vacuo to obtain 3-chloro-5,6-dimethyl-2-(methylsulphanyl)benzoic acid (0.026 g, 36% yield) as a white solid.
[0513] .sup.1H NMR (300 MHZ, CDCl.sub.3) 9.47-8.58 (br s, 1H), 7.34 (s, 1H), 2.40 (s, 3H), 2.28 (s, 3H), 2.27 (s, 3H).
[0514] ES-MS: m/z 229.34 (M1).
[0515] HPLC: Retention time 10.497 min, purity >98% at 280 nm.
Example 9: Synthesis of 3-chloro-2-(difluoromethoxy)-5,6-dimethylbenzoic acid (A-9)
##STR00019##
Step 1: Methyl 3-chloro-2-ethoxy-5,6-xylenecarboxylate
[0516] To a solution of 3-chloro-2-ethoxy-5,6-xylenecarboxylic acid (600 mg, 2.63 mmol) in DMF (5 mL), K.sub.2CO.sub.3 (726 mg, 5.262 mmol, 2 eq.) was introduced, followed by iodomethane (0.5 mL, 7.9 mmol, 3 eq.) and the reaction mixture was stirred at ambient temperature for 18 h. The solvent was removed in vacuo and the residue treated with 2N HCl solution (20 mL) before extraction with CH.sub.2Cl.sub.2 (350 mL). The combined extracts were dried (Na.sub.2SO.sub.4) and concentrated to provide methyl 3-chloro-2-ethoxy-5,6-xylenecarboxylate a greenish oil (620 mg, 97%).
[0517] .sup.1H NMR (300 MHZ, CDCl.sub.3) 7.25 (s, 1H), 4.15 (q, 2H), 3.94 (s, 3H), 2.30 (s, 3H), 2.35 (s, 3H), 1.49 (t, 3H).
Step 2: Methyl 3-chloro-2-hydroxy-5,6-xylenecarboxylate
[0518] To a solution of methyl 3-chloro-2-ethoxy-5,6-xylenecarboxylate (370 mg, 1.525 mmol, 1.0 eq.) in DCM (15 mL) at 0 C., AlCl.sub.3 (811 mg, 6.1 mmol, 4.0 eq.) was added. The reaction mixture was stirred at ambient temperature for 18 h. The reaction was quenched with 2N HCl solution (10 mL) before extraction with CH.sub.2Cl.sub.2 (350 mL). The combined extracts were dried (Na.sub.2SO.sub.4) and concentrated to provide methyl 3-chloro-2-hydroxy-5,6-xylenecarboxylate as a light yellow solid (310 mg, 95%).
[0519] .sup.1H NMR (300 MHZ, CDCl.sub.3) 7.26 (s, 1H), 3.98 (s, 3H), 2.37 (s, 3H), 2.30 (s, 3H).
Step 3: Methyl 3-chloro-2-difluoromethoxy-5,6-xylenecarboxylate
[0520] To a solution of methyl 3-chloro-2-hydroxy-5,6-xylenecarboxylate (230 mg, 1.075 mmol, 1.0 eq.) in CH.sub.3CN (3 mL) at 0 C. was added 6N KOH solution (2.5 mL), followed by dropwise addition of difluoromethoxysulphonyltrifluoromethane (0.344 mL, 1.72 mmol, 1.6 eq.). The reaction mixture was stirred at 0 C. for 10 min. The reaction mixture was quenched with water (20 mL) and extracted with CH.sub.2Cl.sub.2 (350 mL), dried (Na.sub.2SO.sub.4) then concentrated in vacuo to an oil which was purified by silica gel column chromatography eluting with (0-5%) EtOAc/hexane to provide methyl 3-chloro-2-difluoromethoxy-5,6-xylenecarboxylatecolourless oil (210 mg, 80%).
[0521] 1H NMR (300 MHZ, CDCl.sub.3) 7.26 (s, 1H), 6.46 (t, 1H), 3.91 (s, 3H), 2.23 (s, 3H), 2.16 (s, 3H) ppm
[0522] .sup.19F NMR (300 MHZ, CDCl.sub.3) 80.52 ppm.
Step 4: 3-chloro-2-(difluoromethoxy)-5,6-dimethylbenzoic acid
[0523] To a solution of methyl 3-chloro-2-difluoromethoxy-5,6-xylenecarboxylate (85 mg, 0.332 mmol, 1.0 eq.) in MeOH (1 mL) and water (2 mL) 1N solution of NaOH (0.35 mL, 0.354 mmol 1.1 eq.) was added. The mixture was heated in a microwave reactor at 150 C. for 50 min. The solvent was removed in vacuo and the residue was dissolved in water (15 mL), washed with CH.sub.2Cl.sub.2 (310 mL), neutralized with 2N HCl then extracted with CH.sub.2Cl.sub.2 (320 mL). The combined extracts provided 3-chloro-2-(difluoromethoxy)-5,6-dimethylbenzoic acid as a yellow solid (75 mg, 98%).
[0524] .sup.1H NMR (300 MHZ, CD.sub.3OD) 7.04 (s, 1H), 6.46 (t, 1H), 2.13 (s, 6H) ppm
[0525] .sup.19F NMR (300 MHZ, CD.sub.3OD) 81.64 ppm.
[0526] ES-MS: m/z 249.2 (M1).
[0527] HPLC: Retention time 12.18 min, purity 98% at 280 nm.
Example 10: Synthesis of 3-chloro-2-(fluoromethoxy)-5,6-dimethylbenzoic acid (A-10)
##STR00020##
Step 1: 3-Bromo-1-chloro-2-fluoromethoxy-4,5-xylene
[0528] To a solution of 2-bromo-6-chloro-3,4-xylenol (205 mg, 0.872 mmol) in DMF (3 mL), Cs.sub.2CO.sub.3 (567 mg, 1.74 mmol, 2 eq.) was added fluoroiodomethane (418 mg, 2.613 mmol 3.0 eq.) by slow addition. The reaction mixture was stirred at ambient temperature for 18 h, concentrated in vacuo and saturated aqueous NaHCO.sub.3 solution (25 mL) was added and extracted with CH.sub.2Cl.sub.2 (240 mL). The combined extracts were dried (Na.sub.2SO.sub.4), filtered and concentrated in vacuo to provide a white solid (200 mg, 86%).
[0529] .sup.1H NMR (300 MHZ, CDCl.sub.3) 7.26 (s, 1H), 5.72 (s, 1H), 5.54 (s, 1H), 2.30 (s, 3H), 2.25 (s, 3H) ppm.
[0530] .sup.19F NMR (300 MHZ, CDCl.sub.3) 149.23 ppm
Step 2: 3-Chloro-2-(fluoromethoxy)-5,6-dimethylbenzoic acid
[0531] To a solution of 3-bromo-1-chloro-2-fluoromethoxy-4,5-xylene (0.2 g, 0.75 mmol, 1.0 eq.) in THF (5 mL) at 78 C. a solution of 2.5 M n-BuLi in hexane (0.3 mL, 0.75 mmol, 1.0 eq.) was added. The reaction mixture was stirred at this temperature for 45 min. After 50 min. CO.sub.2 (g) was bubbled for 10 min., and the reaction mixture stirred for 1 h at 78 C. and was slowly brought to ambient temperature before quenching with 1M NaOH to pH 11. The mixture was washed with CH.sub.2Cl.sub.2 (20 mL) to remove impurities then the water layer was separated and acidified with 1N HCl to PH 1 followed by extraction with EtOAc (210 mL). The organic layer was dried (Na.sub.2SO.sub.4), filtered, and evaporated to obtain 3-chloro-2-(fluoromethoxy)-5,6-dimethylbenzoic acid (80 mg, 46%) as a white solid.
[0532] .sup.1H NMR (300 MHZ, CD.sub.3OD) 7.35 (s, 1H), 5.72 (s, 1H), 5.54 (s, 1H), 2.30 (s, 3H), 2.25 (s, 3H).
[0533] .sup.19F NMR (300 MHZ, CD.sub.3OD) 149.54
[0534] ES-MS: m/z 231.4 (M1).
[0535] HPLC: Retention time 9.43 min, purity 89% at 280 nm.
Example 11: Synthesis of 3-fluoro-2-methoxy-5,6-dimethylbenzoic acid (A-11)
##STR00021##
Step 1: 3-Bromo-5-fluoro-6-methoxy-2-methylbenzaldehyde
[0536] To a suspension of 1-bromo-5-fluoro-4-methoxy-2-methylbenzene (607 mg, 2.76 mmol) and AgOTf (2.12 g, 8.28 mmol) in dry CH.sub.2Cl.sub.2 (4 mL) a solution of Cl.sub.2CHOMe (0.75 mL, 8.28 mmol) in dry CH.sub.2Cl.sub.2 (2 mL) was added at 78 C. under an argon atmosphere. After stirring at ambient temperature for 16 h, the reaction mixture was quenched with saturated aqueous NaHCO.sub.3. Following further stirring at ambient temperature for 30 min, the reaction mixture was filtered through a pad of celite, the organic layer was separated and the aqueous layer was extracted with CH.sub.2Cl.sub.2 (240 mL). The combined organic layers were washed with brine, dried (Na.sub.2SO.sub.4) and filtered. The filtrate was concentrated in vacuo and the resultant residue was purified by flash column chromatography on silica gel eluting with (0-10%) EtOAc/hexane to afford 3-bromo-5-fluoro-6-methoxy-2-methylbenzaldehyde (479 mg, 70% yield) as off-white solid.
[0537] .sup.1H NMR (300 MHZ, CDCl.sub.3) 10.51 (s, 1H); 7.59 (d, 1H); 4.09 (d, 3H); 2.63 (s, 3H); ES-MS: 248 [M+1].
[0538] .sup.19F NMR (300 MHZ, CDCl.sub.3) 131.16.
Step 2: (3-Bromo-5-fluoro-6-methoxy-2-methylphenyl) methanol
[0539] To a solution of 3-bromo-5-fluoro-6-methoxy-2-methylbenzaldehyde (150 mg, 0.60 mmol) in THF (10 mL) at 0 C. DIBAL-H (1M in toluene, 0.91 mL, 0.91 mmol) was added dropwise over 20 min. The solution was stirred for a further 15 min at 0 C. The solution was allowed to warm to ambient temperature and stirred for 2 h. The reaction was quenched with saturated aqueous sodium potassium tartrate and extracted with EtOAc (250 mL). The combined organic layers were washed with brine (50 mL) dried over (Na.sub.2SO.sub.4), filtered and evaporated. The crude product was purified by chromatography on silica gel (0-10%) EtOAc/hexane to afford (3-bromo-5-fluoro-6-methoxy-2-methylphenyl) methanol (112 mg, 98% yield) as a colourless oil.
[0540] .sup.1H NMR (300 MHZ, CDCl.sub.3) 7.24 (d, 1H); 4.69 (s, 2H); 3.88 (d, 3H); 2.37 (s, 3H); ES-MS: 250 [M+1].
[0541] .sup.19F NMR (300 MHZ, CDCl.sub.3) 131.16.
Step 3: 3-Bromo-5-fluoro-6-methoxy-2-methylbenzoic acid
[0542] To a stirred solution of (3-bromo-5-fluoro-6-methoxy-2-methylphenyl) methanol (155 mg, 0.84 mmol), sodium chlorite (189 mg, 2.1 mmol) in acetonitrile (7 mL) and 1M sodium phosphate buffer (pH6, 7 mL) sodium hypochlorite solution (5 drops, 4-4.99M) was added, followed by TEMPO (6.5 mg, 0.042 mmol). The reaction mixture was stirred at ambient temperature for 2 h. Further sodium hypochlorite (4 drops, 4-4.99M solution) and TEMPO (6.5 mg, 0.042 mmol) were introduced; the addition of sodium hypochlorite and TEMPO was repeated three more times at 8 h intervals. After completion of the reaction, it was cooled to 0 C. and 1M NaOH solution was added to adjust pH 13 and extracted with CH.sub.2Cl.sub.2 (40 mL). The water layer was separated and cooled again to 0 C., acidified with 1M HCl to PH 1, extracted with EtOAc (250 mL), washed with water (210 mL), brine (10 mL), dried (Na.sub.2SO.sub.4), filtered and concentrated to provide 3-bromo-5-fluoro-6-methoxy-2-methylbenzoic acid as a white solid (120 mg, 96%).
[0543] .sup.1H NMR (300 MHZ, CDCl.sub.3) 10.94-10.38 (br, 1H), 7.28 (d, 1H); 3.86 (d, 3H); 2.27 (s, 3H); ES-MS: 363 [M1].
[0544] .sup.19F NMR (300 MHZ, CDCl.sub.3) 130.38.
Step 4: 3-Fluoro-2-methoxy-5,6-dimethylbenzoic acid
[0545] A mixture of 3-bromo-5-fluoro-6-methoxy-2-methylbenzoic acid (50 mg, 0.19 mmol), Pd(dppf)Cl.sub.2 (6.93 mg, 0.0095 mmol), methyl boronic acid (22.6 mg, 0.38 mmol) and potassium carbonate (78.5 mg, 0.57 mmol) in 1,4-dioxane (0.85 mL) and water (0.85 mL) was evacuated and purged with argon three times and the mixture was heated at 90 C. for 12 h then allowed to cool to ambient temperature. Volatiles were removed in vacuo and the crude product was purified by semi-prep HPLC (10 to100% acetonitrile/0.1% formic acid in water) to afford 3-fluoro-2-methoxy-5,6-dimethylbenzoic acid as an off-white solid (25 mg, 96.1%).
[0546] .sup.1H NMR (300 MHZ, CDCl.sub.3) 10.97-10.15 (br, 1H), 6.97 (d, 1H); 3.96 (d, 3H); 2.24 (s, 3H); 2.243 (s, 3H); ES-MS: 197 [M1].
[0547] .sup.19F NMR (300 MHZ, CDCl.sub.3) 133.88.
[0548] HPLC Retention Time: 8.865 min, purity >98% at 280 nm.
Example 12: Synthesis of 3-chloro-2-ethoxy-5,6-dimethylbenzoic acid (A-12)
##STR00022##
Step 1:1-Chloro-2-ethoxy-4,5-xylene
[0549] To a solution of 2-chloro-4,5-xylenol (1.0 g, 6.39 mmol, 1.0 eq.) in DMF (5 mL), K.sub.2CO.sub.3 (1.762 g, 12.77 mmol, 2 eq.) was introduced followed by slow addition of ethyl bromide (1.9 mL, 25.54 mmol, 4 eq.). The reaction mixture was stirred at ambient temperature for 18 h. The solvent was removed in vacuo, saturated NaHCO.sub.3 solution (15 mL) was added to the residue and the mixture extracted with CH.sub.2Cl.sub.2 (320 mL) and the combined extracts dried (Na.sub.2SO.sub.4) and concentrated to afford 1-chloro-2-ethoxy-4,5-xylene as a white solid (1.1 g, 93%).
[0550] .sup.1H NMR (300 MHZ, CDCl.sub.3) 7.27 (s, 1H), 6.71 (s, 1H), 4.10 (q, 2H), 2.35 (s, 3H), 2.30 (s, 3H), 1.41 (t, 3H).
Step 2: 3-Chloro-2-ethoxy-5,6-xylenecarbaldehyde
[0551] To a suspension of 1-chloro-2-ethoxy-4,5-xylene (1.13 g, 6.12 mmol) and AgOTf (4.72 g, 18.36 mmol, 3.0 eq.) in dry CH.sub.2Cl.sub.2 (12 mL) a solution of Cl.sub.2CHOMe (1.75 mL, 18.36 mmol, 3.0 eq.) in dry CH.sub.2Cl.sub.2 (2 mL) was added at 78 C. under an argon atmosphere. After being stirred at ambient temperature for 16 h, the reaction mixture was quenched with saturated aqueous NaHCO.sub.3 (30 mL). After a further 30 min of stirring at ambient temperature the reaction mixture was filtered through a pad of celite. The organic layer was separated, and the aqueous layer was extracted with CH.sub.2Cl.sub.2 (230 mL). The combined organic layers were washed with brine, dried (Na.sub.2SO.sub.4) and filtered. The filtrate was concentrated in vacuo and the resultant residue was purified by flash chromatography on silica gel (0-10%) EtOAc/hexane to afford 3-chloro-2-ethoxy-5,6-xylenecarbaldehyde (970 mg, 86%) as an off-white solid.
[0552] .sup.1H NMR (300 MHZ, CDCl.sub.3) 10.40 (s, 1H), 7.30 (s, 1H), 4.11 (q, 2H), 2.40 (s, 3H), 2.33 (s, 3H), 1.42 (t, 3H).
Step 3: (3-Chloro-2-ethoxy-5,6-xylyl) methanol
[0553] To a solution of 3-chloro-2-ethoxy-5,6-xylenecarbaldehyde (950 mg, 4.47 mmol) in THF (10 mL) at 0 C., DIBAL-H (1M in toluene, 13.4 mL, 13.40 mmol, 3 eq.) was added dropwise over 5 min. The solution was stirred for a further 15 min at 0 C., allowed to warm to ambient temperature and stirred for a further 16 h. The reaction mixture was quenched with saturated aqueous sodium potassium tartrate (20 mL), stirred for 20 min, filtered through pad of celite and extracted with EtOAc (250 mL). The combined organic extracts were washed with brine (50 mL) dried (Na.sub.2SO.sub.4) and evaporated to provide (3-chloro-2-ethoxy-5,6-xylyl) methanol (861 mg, 90%) as a yellow oil.
[0554] .sup.1H NMR (300 MHZ, CDCl.sub.3) 7.18 (s, 1H), 4.80 (d, 2H), 4.04 (q, 2H), 2.27 (s, 3H), 2.31 (s, 3H), 1.46 (t, 3H).
Step 4: 3-Chloro-2-ethoxy-5,6-dimethylbenzoic acid
[0555] To a stirred solution of (3-chloro-2-ethoxy-5,6-xylyl) methanol (766 mg, 3.58 mmol), sodium chlorite (809 mg, 8.92 mmol, 2.5 eq.) in acetonitrile (14 mL) and 1M sodium phosphate buffer (pH6, 14 mL) was added sodium hypochlorite (0.7 mL, 4-4.99M solution) followed by TEMPO (112 mg, 0.716 mmol, 0.2 eq.). The reaction mixture was stirred at ambient temperature for 16 h. After completion of the reaction the mixture was cooled to 0 C. and 1M NaOH solution was added adjusting the pH to 13 and the solution was washed with CH.sub.2Cl.sub.2 (100 mL). The water layer was separated, cooled to 0 C., acidified with 1M HCl to PH 1, extracted with EtOAc (350 mL), and the combined extracts were washed with water (220 mL), brine (10 mL), dried (Na.sub.2SO.sub.4) and evaporated to provide 3-chloro-2-ethoxy-5,6-dimethylbenzoic acid as a white solid (680 mg, 83% yield).
[0556] .sup.1H NMR (300 MHZ, CDCl.sub.3) 7.30 (s, 1H), 4.21 (q, 2H), 2.30 (s, 3H), 2.33 (s, 3H), 1.42 (t, 3H).
[0557] ES-MS: m/z 227.5 (M1).
[0558] HPLC: Retention time 10.384 min, purity 97% at 254 nm.
Example 13: Synthesis of 3-bromo-2-methoxy-5,6-dimethylbenzoic acid (A-13)
##STR00023##
Step 1:1-Bromo-2-methoxy-4,5-dimethylbenzene
[0559] A mixture of 4-methoxy-1,2-dimethylbenzene (1.4 g, 10.28 mmol) and N-bromosuccinimide (1.99 g, 11.2 mmol) in acetonitrile (25 mL) was stirred at ambient temperature for 18 h. The reaction mixture was quenched with water (40 mL) and extracted with EtOAc (2100 mL). The combined organic extracts were washed with saturated brine (50 mL), dried (Na.sub.2SO.sub.4), filtered and evaporated. The crude product was purified by column chromatography on silica gel (0-10%) EtOAc/hexane to afford 1-bromo-2-methoxy-4,5-dimethylbenzene (1.9 g, 86% yield) as an oil.
[0560] .sup.1H NMR (300 MHZ, CDCl.sub.3) 7.17 (s, 1H); 6.78 (s, 1H); 3.92 (s, 3H); 2.29 (s, 3H); 2.23 (s, 3H). ES-MS: 171 [M+1].
Step 2: 3-Bromo-2-methoxy-5,6-dimethylbenzaldehyde
[0561] To a suspension of 1-bromo-2-methoxy-4,5-dimethylbenzene (1 g, 4.65 mmol) and AgOTf (3.58 g, 13.95 mmol) in dry CH.sub.2Cl.sub.2 (8 mL), a solution of Cl.sub.2CHOMe (1.26 mL, 13.95 mmol) in dry CH.sub.2Cl.sub.2 (5 mL) was added at 78 C. under an argon atmosphere. The rection mixture was stirred and gradually allowed to reach ambient temperature over 16 h, the reaction mixture was quenched with saturated aqueous NaHCO.sub.3 then stirring was continued for 30 min. The reaction mixture was filtered through a pad of celite, the organic layer separated and the aqueous layer extracted with CH.sub.2Cl.sub.2 (230 mL). The combined organic extracts were washed with brine, dried (Na.sub.2SO.sub.4) and filtered. The filtrate was concentrated in vacuo and the resultant residue was purified by flash column chromatography on silica gel (0-10%) EtOAc/hexane to afford 3-bromo-2-methoxy-5,6-dimethylbenzaldehyde (553 mg, 49% yield) as a solid.
[0562] .sup.1H NMR (300 MHZ, CDCl.sub.3) 10.55 (s, 1H); 7.60 (s, 1H); 3.95 (s, 3H); 2.48 (s, 3H); 2.32 (s, 3H). ES-MS: 245 [M+1].
Step 3: 3-Bromo-2-methoxy-5,6-dimethylphenyl) methanol
[0563] To a solution of 3-bromo-2-methoxy-5,6-dimethylbenzaldehyde (366 mg, 1.51 mmol) in THF (15 mL) at 0 C., DIBAL-H (1M in toluene, 2.26 mL, 2.26 mmol) was introduced dropwise over 20 min. The solution was stirred for 15 min at 0 C. and was allowed to warm to ambient temperature and stirred for a further 2 h. The reaction was quenched with saturated aqueous sodium potassium tartrate solution (10 mL) and extracted with EtOAc (250 mL). The combined organic layers were washed with brine (50 mL), dried (Na.sub.2SO.sub.4) filtered and evaporated. The crude residue was purified by chromatography on silica gel (0-10%) EtOAc/hexane to afford (3-bromo-2-methoxy-5,6-dimethylphenyl) methanol (350 mg, 95% yield) as a colourless oil.
[0564] .sup.1H NMR (300 MHZ, CDCl.sub.3) 7.37 (s, 1H); 4.83 (s, 2H); 3.92 (s, 3H); 2.32 (s, 3H); 2.28 (s, 3H). ES-MS: 246 [M+1].
Step 4: 3-Bromo-2-methoxy-5,6-dimethylbenzoic acid
[0565] To a stirred solution of (3-bromo-2-methoxy-5,6-dimethylphenyl) methanol (350 mg, 1.43 mmol), sodium chlorite (323 mg, 3.58 mmol) in acetonitrile (12 mL) and 1M sodium phosphate buffer (pH6, 12 mL) was added sodium hypochlorite (5 drops, 4-4.99M solution) followed by TEMPO (11.1 mg, 0.0715 mmol). The reaction mixture was stirred at ambient temperature for 2 h. Further sodium hypochlorite (4 drops, 4-4.99M solution) and TEMPO (11.1 mg, 0.0715 mmol) were added and addition of this sodium hypochlorite/TEMPO reagent took place three more times at 8 h intervals. After completion of the reaction, the mixture was cooled to 0 C. and 1M NaOH solution was added to adjust the pH to 13 before being extracted with CH.sub.2Cl.sub.2 (40 mL). The water layer was separated and cooled again to 0 C., acidified with 1M HCl to PH 1, extracted with EtOAc (250 mL). The combined extracts were washed with water (210 mL), brine (10 mL), dried (Na.sub.2SO.sub.4). filtered and concentrated to provide 3-bromo-2-methoxy-5,6-dimethylbenzoic acid as a white solid (455 mg, 89%).
[0566] .sup.1H NMR (300 MHZ, CDCl.sub.3) 11.91-11.57 (br, 1H), 7.44 (s, 1H); 3.99 (s, 3H); 2.29 (s, 3H); 2.28 (s, 3H).
[0567] ES-MS: 258 [M1].
[0568] HPLC Retention Time: 9.554 min, >98% purity at 280 nm.
Example 14: Synthesis of 3-chloro-2-methoxy-5,6-dimethylbenzoic acid (A-14)
##STR00024##
Step 1:1-Chloro-2-methoxy-4,5-dimethylbenzene
[0569] To a solution of 2-chloro-4,5-dimethylphenol (3.07 g, 19.6 mmol) and K.sub.2CO.sub.3 (5.41 g, 39.2 mmol) in DMF (25 mL) methyl Iodide (2.44 mL, 39.2 mmol) was added and the resulting solution was stirred at ambient temperature for 18 h. The reaction mixture was quenched with water (40 mL) and extracted with EtOAc (2100 mL). The combined organic extracts were washed with brine (50 mL), dried (Na.sub.2SO.sub.4), filtered and evaporated. The crude product was purified by column chromatography on silica gel (0-10%) EtOAc/hexane to afford 1-chloro-2-methoxy-4,5-dimethylbenzene (3.31 g, 98% yield) as a colourless oil.
[0570] .sup.1H NMR (300 MHZ, CDCl.sub.3) 7.17 (s, 1H); 6.78 (s, 1H); 3.92 (s, 3H); 2.29 (s, 3H); 2.23 (s, 3H). ES-MS: 171 [M+1].
Step 2: 3-Chloro-2-methoxy-5,6-dimethylbenzaldehyde
[0571] To a suspension of 1-chloro-2-methoxy-4,5-dimethylbenzene (1.4 g, 8.2 mmol) and AgOTf (6.3 g, 24.6 mmol) in dry CH.sub.2Cl.sub.2 (12 mL), a solution of Cl.sub.2CHOMe (2.22 mL, 24.6 mmol) in dry CH.sub.2Cl.sub.2 (6 mL) was added at 78 C. under an argon atmosphere. After stirring at ambient temperature for 16 h, the reaction mixture was quenched with saturated aqueous NaHCO.sub.3. Following further stirring at ambient temperature for 30 min, the reaction mixture was filtered through a pad of celite. The organic layer was separated and the aqueous layer extracted with CH.sub.2Cl.sub.2 (250 mL). The combined organic layers were washed with brine (50 mL), dried (Na.sub.2SO.sub.4) and filtered. The filtrate was concentrated and the resultant residue was purified by flash column chromatography on silica gel (0-10%) to afford 3-chloro-2-methoxy-5,6-dimethylbenzaldehyde (1.2 g, 86% yield) as a white solid.
[0572] .sup.1H NMR (300 MHZ, CDCl.sub.3) 10.54 (s, 1H); 7.41 (s, 1H); 3.95 (s, 3H); 2.48 (s, 3H); 2.28 (s, 3H). ES-MS: 199 [M+1].
Step 3: (3-Chloro-2-methoxy-5,6-dimethylphenyl) methanol
[0573] To a solution of 3-chloro-2-methoxy-5,6-dimethylbenzaldehyde (565 mg, 2.84 mmol) in THF (17 mL) at 0 C., DIBAL-H (1M in toluene, 4.26 mL, 4.26 mmol) was added dropwise over 20 min. The solution was stirred for 15 min at 0 C. and allowed to warm to ambient temperature and stirred for a further 2 h. The reaction was quenched with saturated aqueous sodium potassium tartrate and extracted with EtOAc (250 mL). The combined organic extracts were washed with brine (50 mL), dried (Na.sub.2SO.sub.4), filtered and evaporated. The crude residue was purified by chromatography on silica gel (0-10%) EtOAc/hexane to afford (3-chloro-2-methoxy-5,6-dimethylphenyl) methanol (475 mg, 83% yield) as a colourless oil.
[0574] .sup.1H NMR (300 MHZ, CDCl.sub.3) 7.23 (s, 1H); 4.84 (s, 2H); 3.92 (s, 3H); 2.34 (s, 3H); 2.29 (s, 3H).
[0575] ES-MS: 201 [M+1].
Step 4: 3-Chloro-2-methoxy-5,6-dimethylbenzoic acid
[0576] To a stirred solution of (3-chloro-2-methoxy-5,6-dimethylphenyl) methanol (475 mg, 2.37 mmol), sodium chlorite (535 mg, 5.93 mmol) in acetonitrile (7 mL) and 1M sodium phosphate buffer (pH6, 7 mL), sodium hypochlorite (5 drops, 4-4.99M solution) were added followed by TEMPO (18.4 mg, 0.118 mmol). The reaction mixture was stirred at ambient temperature for 2 h. Further sodium hypochlorite (4 drops, 4-4.99M solution) plus TEMPO (18.4 mg, 0.118 mmol) were added and addition of this sodium hypochlorite/TEMPO reagent took place three more times at 8 h intervals. After completion, the reaction mixture was cooled to 0 C. and 1M NaOH solution was added to adjust pH 13 and extracted with CH.sub.2Cl.sub.2 (40 mL). The water layer was separated and cooled again to 0 C., acidified with 1M HCl to PH 1, extracted with EtOAc (250 mL), washed with water (210 mL), brine (10 mL), dried (Na.sub.2SO.sub.4), filtered and concentrated in vacuo to provide 3-chloro-2-methoxy-5,6-dimethylbenzoic acid as a white solid (455 mg, 89%).
[0577] .sup.1H NMR (300 MHZ, CDCl.sub.3) 10.94-10.38 (br, 1H), 7.32 (s, 1H); 3.99 (s, 3H); 2.33 (s, 3H); 2.31 (s, 3H).
[0578] ES-MS: 215 [M1].
[0579] HPLC Retention Time: 9.683 min, purity >98% at 280 nm.
Example 15: Electrophysiological Measurement of Compound Inhibition of CIC-1 in Rat Muscle
[0580] The investigatory goal of these experiments was to evaluate whether compounds inhibit CIC-1 channels in native tissue of rat skeletal muscle fibres. Apparent CIC-1 affinity was reported by the concentration of compound at which 50% of the compound's full inhibition of CIC-1 was observed (EC.sub.50).
[0581] Experimentally, G.sub.m was measured in individual fibres of whole rat soleus muscles using a three micro-electrodes technique described in this example and in full detail elsewhere (Riisager et al., Determination of cable parameters in skeletal muscle fibres during repetitive firing of action potentials. Journal of Physiology, 2014, 592, 4417-4429). Briefly, intact rat soleus muscles were dissected out from 12-14 week old Wistar rats and placed in an experimental chamber that was perfused with a standard Krebs Ringer solution containing 122 mM NaCl, 25 mM NaHCO.sub.3, 2.8 mM KCl, 1.2 mM KH.sub.2PO.sub.4, 1.2 mM MgSO.sub.4, 1.3 mM CaCl.sub.2), 5.0 mM D-glucose. During experiments, the solution was kept at approx. 30 C. and continuously equilibrated with a mixture of 95% O.sub.2 and 5% CO.sub.2, PH 7.4. The experimental chamber was placed in Nikon upright microscope that was used to visualize individual muscle fibres and the three electrodes (glass pipettes filled with 2 M potassium citrate). For G.sub.m measurements, the electrodes were inserted into the same fibre with known inter-electrode distances of 0.35-0.5 mm (V1-V2, X1) and 1.1-1.5 mm (V1-V3, X3). The membrane potential of the impaled muscle fibre was recorded by all electrodes. Two of the electrodes were furthermore used to inject 50 ms current pulses of 30 nA. Given the positions of the electrodes, three different inter-electrode distances could be identified (X1-X2, X1-X3, X2-X3) and hence the membrane potential responses to the current injections could be obtained at three distances from the point of current injection. The steady state voltage deflection at each distance was divided by the magnitude of current injected (30 nA) and the resulting transfer resistances were plotted against inter-electrode distance and the data was fitted to a mono-exponential function from which G.sub.m could be calculated using linear cable theory.
[0582] To establish a dose response relationship, G.sub.m was first determined in 10 muscle fibres in the absence of compound and then at four increasing compound concentrations with G.sub.m determinations in 5-10 fibres at each concentration. The average G.sub.m values at each concentration were plotted against compound concentration and the data was fitted to sigmoidal function to obtain an EC.sub.50 value. Table 1 shows the EC.sub.50 values for a range of compounds with n values referring to number of experiments that each reflect recordings from around 50 fibres.
TABLE-US-00001 TABLE 1 Inhibition of CIC-1 ion channel using compounds of the disclosure Compound investigated EC.sub.50 (M) A-3 7.4 A-4 1.7 A-5 6.6 A-6 5.8 A-7 5.3 A-8 3.2 A-9 4.3 A-10 4.1 A-13 4.4 A-14 3.2
Example 16: Measurement of Force in an In Vitro Model
[0583] The current disclosure relates to compounds that inhibit CIC-1 ion channels and increase muscle excitability and thereby improve muscle function in clinical conditions where muscle activation is failing. Such conditions result in loss of contractile function of skeletal muscle, weakness and excessive fatigue. In this series of experiments the compounds were tested for their ability to restore contractile function of isolated rat muscle when the neuromuscular transmission had been compromised akin to neuromuscular disorders.
[0584] Experimentally, soleus muscles from 4-5 week old rats were isolated with the motor nerve remaining attached. The nerve-muscle preparations were mounted in experimental setups that enabled electrical stimulation of the motor nerve. Stimulation of the motor nerve led to activation of the muscle fibres and ensuing force production that was recorded. The nerve-muscle preparations were also in these experiments incubated in the standard Krebs Ringer (see example 15) and the solution was heated to 30 C. and continuously equilibrated with a mixture of 95% O.sub.2 and 5% CO.sub.2, PH 7.4.
[0585] After mounting the nerve-muscle preparation in the experimental setup, the contractile function of the muscle was initially assessed under the control conditions. Sub-maximal concentration of tubocurarine (115 nM), an acetylcholine receptor antagonist, was then added to the experimental bath to impose partial inhibition of the ability of the motor nerve to activate the muscle fibres. The experimental condition mimics the failing neuromuscular transmission in a range of neuromuscular disorders. After addition of tubocurarine the contractile force declined over the next 90 minutes to 10-50% of the control force. The test compound was then added to obtain the required compound concentration in the bath and the contractile force recovered was measured. To quantify the ability of the compound to restore force the percentage of the initial force that was restored was determined after 40 mins of compound exposure and the point increase is reported in Table 2.
TABLE-US-00002 TABLE 2 Percentage increase of initial force that was restored Concentration of test Compound investigated compound (M) Point increase (%) A-1 50 19 A-2 100 16 A-3 50 46 A-4 10 23 A-5 50 48 A-6 10 28 A-7 50 44 A-8 10 24 A-9 50 50 A-10 10 21 A-11 50 25 A-12 50 39 A-13 10 29 A-14 50 44
[0586] In conclusion, this example demonstrates that the compounds of the present disclosure are able to increase muscle excitability and thereby improve muscle function in clinical conditions.
Example 5: Measurement of In Situ Muscle Contractile Characteristics
[0587] Isometric hindlimb force was measured in 12-week old female Lewis rats in the presence and absence of compound.
[0588] Rats were placed under anaesthesia with isoflurane (2-4%), intubated and subsequently connected to a micro ventilator (Microvent 1, Hallowell EMC, US). Two stimulation electrodes were inserted through the skin to stimulate the sciatic nerve. A small incision was made proximal to the ankle, to expose the Achilles tendon, which was tied by cotton string, and connected to a force transducer (Fort250, World Precision Instruments) with adjustable position (Vernier control). The Achilles tendon was then cut distal to the attached cotton string. The rat was placed on a heated pad, and to prevent movement artefacts from contraction of the ankle dorsiflexors, the foot was fixated by tape on a footplate.
[0589] Muscle contractile properties were assessed by applying an electrical current (under supramaximal voltage conditions) to the nerve and recording the force generated by the muscle. The muscle was stretched until maximal force was obtained, when assessed by 2 Hz stimulation. Isometric force was measured every 30 seconds at 12 Hz (Twitch), 10 pulses, and at every 5 minutes at 80 Hz (Tetanic) for 1 second (80 pulses). This stimulation pattern was employed throughout the experiment, expect in few cases where 80 Hz stimulation was replaced by 12 Hz (10 pulses). Neuromuscular transmission was partially inhibited by constant infusion of Cisatracurium (Nimbex, GlaxoSmithKline) at a concentration of 0.1 mg/kg at an adjustable infusion speed, adjusted individually for each animal to obtain a level of inhibition of ca. 50% of the forced generated at 12 Hz stimulation on the 4th pulse. When the level of neuromuscular inhibition was stable, the test article (sodium salt of test article dissolved in PBS) was injected i.v. or p.o. at the chosen concentration. The effect of test article was assessed on its ability to increase force generated from the stimulation pattern applied. The effect was assessed in the ability to increase force per se (tetanic, 80 Hz, stimulation), and the ratio between individual twitch peaks (12 Hz stimulation). The effect was monitored for at least 1 hour after injection of test article. In addition, the time from injection of test article to maximal effect on force (both twitch and tetanic) was noted and the time for the effect to disappear (return to baseline), if possible. When appropriate the infusion of neuromuscular blocking agent was ceased, with the stimulation pattern continued, and the return of force to control levels was monitored. Animals were sacrificed by cervical dislocation while still fully sedated.
[0590] The sodium salt of compound A-7 was dosed 2.5 mg/kg i.v. in PBS resulting in an increase in tetanic force of 27%.
[0591] The sodium salt of compound A-14 was dosed 10 mg/kg p.o. in PBS resulting in an increase in tetanic force of 45%.
[0592] This demonstrates that compounds of the disclosure can restore force to muscles in vivo which have been partially inhibited by a neuromuscular blocker.