Treatment of tinnitus through modulation of chloride co-transporter nkcc1 in the auditory system

Abstract

The present invention relates to the treatment of prevention of tinnitus. More precisely, the present invention relates to a compound modulating chloride co-transporter NKCC1 (chloride co-transporter modulator) for use in the treatment of tinnitus. In addition, the present invention concerns pharmaceutical composition comprising such an NKCC1 chloride co-transporter modulator as an active agent a method for the treatment or prevention of tinnitus by administering such a chloride co-transporter modulator, and a screening method for the identification and characterization of compounds capable of modulation chloride co-transporter NKCC1.

1 . A method for treating or preventing tinnitus in a subject in need thereof comprising administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of a compound that modulates sodium potassium chloride co-transporter 1 (NKCC1, wherein the compound decreases NKCC1 expression or activity in auditory sensorineural structures in the subject following administration. 
     
     
         2 . (canceled) 
     
     
         3 . (canceled) 
     
     
         4 . The method of  claim 1 , wherein the therapeutically effective amount of the compound is an amount sufficient to increase activity and/or expression of KCC2 in auditory sensorineural structures in the subject. 
     
     
         5 . (canceled) 
     
     
         6 . The method of  claim 1 , wherein the compound is selected from the group consisting of thiazides and sulfonamides. 
     
     
         7 . The method of  claim 6 , wherein the compound is a sulfonamide selected from the group consisting of acetazolamide, azosemide, bumetanide, chlorthalidone, clopamide, furosemide, hydrochlorothiazide (HCT, HCTZ, HZT), indapamide, mefruside, metolazone, piretanide, tripamide xipamide, dichlorphenamide (DCP), dorzolamide, ethoxzolamide, sultiame, or zonisamide or analogs thereof. 
     
     
         8 . The method of  claim 6 , wherein the sulfonamide compound is a diuretic compound selected from the group consisting of bumetanide, furosemide, piretanide, azosemide, and torsemide or analogs thereof. 
     
     
         9 . The method of  claim 8 , wherein diuretic compound is selected from the group consisting of
 (i) bumetanide, bumetanide aldehyde, bumetanide methyl ester, bumetanide cyanomethyl ester, bumetanide ethyl ester, bumetanide isoamyl ester, bumetanide octyl ester, bumetanide benzyl ester, bumetanide dibenzylamide, bumetanide diethylamide, bumetanide morpholinoethyl ester, bumetanide 3-(dimethylaminopropyl) ester, bumetanide N,N-diethyl glycolamido ester, bumetanide N,N-dimethylglycolamido ester, bumetanide pivaxetil ester, bumetanide propaxetil ester, bumetanide methoy(polyethyleneoxy) n-1 -ethyl ester, bumetanide benzyltrimethylammonium salt and bumetanide cetyl-trimethylammonium salt, bumetanide [—(C═O)—SH] thioacid, bumetanide S-methyl thioester, bumetanide S-cyanotnethyl thioester, bumetanide S-ethyl thioester, bumetanide S-isoantyl thioester, bumetanide S-octyl thioester, bumetanide S-benzyl thioester, bumetanide S-(morpholinoethyl) thioester, bumetanide S-[3-(dimethylaminopropyl)] thioester, bumetanide S—N,N-diethylglycolamido) thioester, bumetanide S—(N,N-dimethylglycolamido) thioester, bumetanide S-pivaxetil thioester, bumetanide S-propaxetil thioester, bumetanide S-[methoxy(polyethyleneoxy) n-1 -ethyl] thioester, bumetanide [—(C═O)—S − ] benzyltrimethyl-ammonium thioacid salt and bumetanide [—(C═O)—S − ] cetyltrimethylammonium thioacid salt, metastable bumetanide [—(C═S)—OH] thioacid, bumetanide O-methyl thioester, bumetanide O-cyanomethyl thioester, bumetanide O-ethyl thioester, bumetanide O-isoamyl thioester, bumetanide O-octyl thioester, bumetanide O-benzyl thioester, bumetanide O—(morpholinoethyl) thioester, bumetanide O—[3-(dimethylaminopropyl)] thioester, bumetanide O—(N,N-diethylglycolamido) thioester, bumetanide, O—(N,N-dimethylglycolamido) thioester, bumetanide O-pivaxetil thioester, bumetanide O-propaxetil thioester, bumetanide O-[methoxy(poryethyleneoxy) n-1 -ethyl] thioester, bumetanide [—(C═S)—O − ] benzyltrimemyl-ammonium thioacid salt and bumetanide [—(C═S)—O − ] cetyltrimethylammonium thioacid salt, bumetanide thioaldehyde, bumetanide [—(C═S)—SH] dithioacid, bumetanide methyl dithioester, bumetanide cyanomethyl dithioester, bumetanide ethyl dithioester, bumetanide isoamyl dithioester, bumetanide octyl dithioester, bumetanide benzyl dithioester, bumetanide dibenzylthioamide, bumetanide diethylthioamide, bumetanide morpholinoethyl dithioester, bumetanide 3-(dimethylaminopropyl) dithioester, bumetanide N,N-diethylglycolamido dithioester, bumetanide N,N-dimethylglycolamido dithioester, bumetanide pivaxetil dithioester, bumetanide propaxetil dithioester, bumetanide methoxy(polyethyleneoxy) n-1 -ethyl dithioester, bumetanide benzyltrimethyl ammonium dithioacid salt and bumetanide cetyltrimethylamnionium dithioacid salt and   (ii) furosemide, furosemide aldehyde, furosemide methyl ester, furosemide cyanomethyl ester, furosemide ethyl ester, furosemide isoamyl ester, furosemide octyl ester, furosemide benzyl ester, furosemide morpholinoethyl ester, furosemide 3-(dimethylaminopropyl) ester, furosemide N,N-diethylglycolamido ester, furosemide N,N-dimethylglycolamido ester, furosemide pivaxetil ester, furosemide propaxetil ester, furosemide methoxy(polyethyleneoxy) n-1 -ethyl ester, furosemide benzyltrimethylammonium acid salt and furosemide cetyltrimethylammonium acid salt and   (iii) piretanide, piretanide aldehyde piretanide methyl ester, piretanide cyanomethyl ester, piretanide ethyl ester, piretanide isoamyl ester, piretanide octyl ester, piretanide benzyl ester, piretanide dibenzylamide, piretanide diethylamide, piretanide morpholinoethyl ester, piretanide 3-(dimethylaminopropyl) ester, piretanide N,N-diethylglycolamide ester, piretanide dimethylglycolamide ester, piretanide pivaxetil ester, piretanide propaxetil ester, piretanide methoxy(polyethyleneoxy) n-1 -ethyl ester, piretanide benzyltrimethylammonium salt and piretanide cetyltrimethylammonium salt and   (iv) tetrazolyl-substituted azosemides, in particular methoxymethyl tetrazolyl-substituted azosemides, methylthiomethyl tetrazolyl-substituted azosemides and N-mPEG350-tetrazolyl-substituted azosemides), azosemide benzyltrimethylammonium salt and/or azosemide cetyltrimethylammonium salt and   (v) pyridine-substituted torsemide quaternary ammonium salts or the corresponding inner salts (zwitterions), in particular methoxymethyl pyridinium torsemide salts, methylthiomethyl pyridinium torsemide salts and N-mPEG350-pyridinium torsemide salts.   
     
     
         10 . The method of  claim 6 , wherein the thiazide compound is selected from the group consisting of bendroflumethiazide, benzthiazide, chlorothiazide, hydrochlorothiazide, hydroflumethiazide, methylclothiazide, polythiazide, trichlor-methiazide, chlorthalidone, indapamide, metolazone or quinethazone or analogs thereof. 
     
     
         11 . The method of  claim 1 , further comprising administering to the subject one or more carbonic anhydrase inhibitors selected from acetazomalide, dichlorphenamide, dorzolamide, brinzolamide and/or methazolamide. 
     
     
         12 . The method of  claim 1 , wherein the compound is an antibody, an antibody fragment or a humanized antibody antagonizing NKCC1 activity or an RNA selected from the group consisting of an siRNA, shRNA and anti-sense RNA that reduces, NKCC1 expression. 
     
     
         13 . The method of  claim 1 , wherein the compound is selected from CCC interacting protein (CIP1) or a functional peptide derived therefom, N-ethyimaleimide (NEM) or staurosporin. 
     
     
         14 . The method of  claim 1 , further comprising administering to the subject one or more GABAergic agonists and/or one or more glycine agonists. 
     
     
         15 . (canceled) 
     
     
         16 . (canceled) 
     
     
         17 . The method of  claim 1 , wherein the pharmaceutical composition is provided in a liquid, semi-liquid or viscous form, preferably in a gel-like form. 
     
     
         18 . The method of  claim 1 , wherein the pharmaceutical composition contains a biodegradable polymer, selected from the group consisting of hyaluronic acid resp. hyaluronates, lecithin gels, (poly)alanine derivatives, pluronics, poly(ethyleneglycol), poloxamers, chitosans, xyloglucans, collagens, fibrins, polyesters, poly(lactides), poly(glycolide) or their co-polymers PLGA, sucrose acetate isobutyrate, and glycerol monooleate. 
     
     
         19 . (canceled) 
     
     
         20 . The method of  claim 1 , wherein the pharmaceutical composition is administered locally. 
     
     
         21 . The method of  claim 1 , wherein the pharmaceutical composition is administered to the middle/inner ear interface, preferably to the round and/or oval window membrane. 
     
     
         22 - 24 . (canceled) 
     
     
         25 . A screening method for the identification and characterization of compounds capable of modulating one or more chloride transporters comprising, (a) providing cells stably expressing the one or more chloride transporters, (b) adding a test compound to the cells, (c) adding a transporter cation and (d) measuring the cation transport across the cell membrane. 
     
     
         26 . (canceled) 
     
     
         27 . The screening method according to  claim 25 , wherein the one or more chloride transporters are selected from NKCC1 and KCC2.