Rescue of gonadotropin releasing hormone receptor mutants

Abstract

Herein disclosed is a method of rescuing gonadotropin-releasing hormone receptor (GnRHR) mutants with IN3 or a mimetic thereof. IN3 significantly rescues 11 missense mutations as assessed by radioligand binding and by IP production. Such rescue occurred despite widely disparate loci along the receptor. In addition, many altered GnRH receptors (terminally truncated, internal deletions, or lacking the ability to form bridges to form tertiary structure) were rescued with IN3.

1 . An in vivo method of rescuing a mutant gonadotropin-releasing hormone receptor (GnRHR), comprising contacting a cell expressing the mutant GnRHR with a therapeutic amount of IN3. 
     
     
         2 . The method of  claim 1 , wherein the GnRHR mutant includes a N 10 K, T 32 I, E 90 K, Q 106 R, A 129 D, R 139 H, C 200 Y, R 262 Q, L 266 R, C 279 Y, or Y 284 C mutation. 
     
     
         3 . The method of  claim 1 , wherein the GnRHR mutant includes a N 10 K, T 32 I, Q 106 R, R 262 Q, or Y 284 C mutation. 
     
     
         4 . The method of  claim 1 , wherein the GnRHR mutant includes an E 90 K mutation. 
     
     
         5 . The method of  claim 1 , wherein the GnRHR mutant does not include a S 168 R or a S 217 R mutation. 
     
     
         6 . The method of  claim 1 , wherein contacting the cell with a therapeutic amount of IN3 comprises administering IN3 to a subject having hypogonadotropic hypogonadism (HH). 
     
     
         7 . The method of  claim 6 , wherein in the subject is a human. 
     
     
         8 . The method of  claim 1 , wherein rescue of the mutant GnRHR results in an at least 25% increased binding of the mutant receptor to buserelin, when compared to binding in the absence of IN3. 
     
     
         9 . The method of  claim 1 , wherein rescue of the mutant GnRHR results in an at least 25% increased GnRH agonist-stimulated inositol phosphate (IP) production, when compared to IP production in the absence of IN3. 
     
     
         10 . The method of  claim 1 , wherein rescue of the mutant GnRHR results in an at least a 25% increase in surface-bound receptor-ligand complex internalized, when compared to surface-bound receptor-ligand complex internalized in the absence of IN3. 
     
     
         11 . The method of  claim 1 , wherein the IN3 is present in a pharmaceutically acceptable carrier. 
     
     
         12 . The method of  claim 6 , wherein the subject is also administered a steroid. 
     
     
         13 . The method of  claim 6 , wherein the subject is also administered a gonadotropin. 
     
     
         14 . A method of reducing symptoms associated with HH, comprising administering a therapeutic amount of IN3 to a subject having HH. 
     
     
         15 . The method of  claim 14 , wherein the symptoms are reduced by 25%. 
     
     
         16 . An in vivo method of rescuing a mutant gonadotropin-releasing hormone receptor (GnRHR), comprising contacting a cell expressing the mutant GnRHR with a mimetic of IN3 and rescue of the mutant GnRHR results in an at least 25% increased binding of the mutant receptor to buserelin, an at least 25% increased GnRH agonist-stimulated inositol phosphate (IP) production, or an at least a 25% increase in surface-bound receptor-ligand complex internalized, when compared to an absence of the mimetic of IN3. 
     
     
         17 . The method of  claim 16 , wherein the mimetic of IN3 comprises [(2S)-N-[2-(4-carboxyphenyl)ethyl]-2-[5-[1,1-dimethyl-2-(4-methylpiperazin-1-yl)-2-oxoethyl]-2-(3,5-dimethylphenyl)-1H-indol-3-yl]propan-1-aminium trifluoroacetate] or [(2S)-2-[5-[2-(2-azabicyclo[2.2.2]oct-2-yl)-1,1-dimethyl-2-oxoethyl]-2-(3,5-dimethylphenyl)-1H-indol-3-yl]-N-{2-[4-(methyl-sulfinyl)phenyl]ethyl}propan-1-amine]. 
     
     
         18 . The method of  claim 16 , wherein the GnRHR mutant includes a N 10 K, T 32 I, E 90 K, Q 106 R, A 129 D, R 139 H, C 200 Y, R 262 Q, L 266 R, C 279 Y, or Y 284 C mutation. 
     
     
         19 . The method of  claim 16 , wherein the GnRHR mutant includes a N 10 K, T 32 I, Q 106 R, R 262 Q, or Y 284 C mutation. 
     
     
         20 . The method of  claim 16 , wherein the GnRHR mutant includes an E 90 K mutation.