Antibody Fragments Against the Insulin Receptor and Uses Thereof to Treat Hypoglycemia

Abstract

The present disclosure relates, in general, to methods of treating or preventing hypoglycemia using a negative modulator antibody fragment that binds to the insulin receptor and modulates the action of insulin at the insulin receptor.

1 . An antibody or fragment thereof comprising three heavy chain CDRs having the amino acid sequence set out in SEQ ID NOs: 5-7, 8-10 and 50-112 and three light chain CDRs have the amino acid sequences set out in SEQ ID NOs: 14-16 or SEQ ID NOs: 20-28 wherein the antibody or fragment thereof binds to i) insulin receptor or (ii) a complex comprising insulin and insulin receptor, or both (i) and (ii). 
     
     
         2 . An antibody or fragment thereof of  claim 1  wherein the heavy chain variable region amino acid sequence is set out in SEQ ID NOs: 1, 2 or 29-49, and the light chain variable region amino acid sequence is set out in SEQ ID NOs: 4 or 17-19. 
     
     
         3 . An antibody or fragment thereof comprising three heavy chain CDRs having the amino acid sequence set out in SEQ ID NOs: 8-10 and three light chain CDRs have the amino acid sequences set out in SEQ ID NOs: 14-16, wherein the antibody or fragment thereof binds to i) insulin receptor or (ii) a complex comprising insulin and insulin receptor, or both (i) and (ii). 
     
     
         4 . The antibody or fragment thereof of  claim 3  wherein the heavy chain variable region amino acid sequence is set out in SEQ ID NO: 2 and the light chain variable region amino acid sequence is set out in SEQ ID NO: 4. 
     
     
         5 . An antibody fragment comprising three heavy chain CDRs having the amino acid sequences set out in SEQ ID NOs: 5-7 or 8-10 and three light chain CDRs having the amino acid sequences set out in SEQ ID NOs: 11-13 or 14-16, wherein the antibody fragment binds to i) insulin receptor or (ii) a complex comprising insulin and insulin receptor, or both (i) and (ii). 
     
     
         6 . The antibody fragment of  claim 3  wherein the heavy chain variable region amino acid sequence is set out in SEQ ID NO: 1 or 2 and the light chain variable region amino acid sequence is set out in SEQ ID NO: 3 or 4. 
     
     
         7 . The antibody fragment of  claim 5  wherein the three heavy chain CDRs have the amino acid sequences set out in SEQ ID NOs: 8-10 and three light chain CDRs have the amino acid sequences set out in SEQ ID NOs: 14-16. 
     
     
         8 . The antibody fragment of  claim 5  wherein the heavy chain variable region amino acid sequence is set out in SEQ ID NO: 2 and the light chain variable region amino acid sequences is set out in SEQ ID NO: 4. 
     
     
         9 . The antibody fragment of  claim 1  which is a Fab fragment. 
     
     
         10 . The antibody fragment of  claim 1  wherein the antibody or antibody fragment binds to (i) insulin receptor or (ii) a complex comprising insulin and insulin receptor, or both (i) and (ii), with an equilibrium dissociation constant KD of 10-5M or less that is capable of weakening the binding affinity between insulin and insulin receptor by at least about 1.5-fold, optionally up to 1000-fold. 
     
     
         11 . (canceled) 
     
     
         12 . The antibody or antibody fragment of  claim 1 , wherein the antibody increases the EC50 of insulin signaling activity by about 2-fold to 1000-fold, optionally in a pAKT assay. 
     
     
         13 . (canceled) 
     
     
         14 . The antibody or antibody fragment of  claim 1 , wherein said antibody is a human antibody. 
     
     
         15 . (canceled) 
     
     
         16 . A method of preparing a sterile pharmaceutical composition, comprising adding a sterile pharmaceutically acceptable diluent to an antibody of  claim 1 . 
     
     
         17 . A sterile composition comprising the antibody of  claim 1  and a sterile pharmaceutically acceptable diluent. 
     
     
         18 . A method of treating hypoglycemia, comprising administering to a subject in need thereof an antibody fragment according to  claim 1  that is a negative modulator of insulin binding to the insulin receptor and/or insulin action at the insulin receptor in an amount effective to ameliorate hypoglycemia. 
     
     
         19 . The method of  claim 18 , wherein the hypoglycemia is selected from the group consisting of sulfonylurea-induced hypoglycemia, insulin-induced hypoglycemia, nocturnal hypoglycemia, hypoglycemia following post-bariatric surgery, and hypoglycemia in subjects with inherited metabolic and insulin sensitivity disorders. 
     
     
         20 . A method for treating or preventing nocturnal hypoglycemia comprising administering to a subject in need thereof an antibody fragment according to  claim 1  that is a negative modulator of insulin binding to the insulin receptor and/or insulin action at the insulin receptor in an amount effective to ameliorate nocturnal hypoglycemia. 
     
     
         21 . The method of  claim 18 , wherein the antibody fragment binds to (i) insulin receptor or (ii) a complex comprising insulin and insulin receptor, or both (i) and (ii), with an equilibrium dissociation constant KD of 10-5 M or less that is capable of weakening the binding affinity between insulin and insulin receptor by at least about 1.5-fold, optionally up to 1000-fold. 
     
     
         22 . The method of  claim 21 , wherein the antibody fragment increases the EC50 of insulin signaling activity by about 2-fold to 1000-fold, optionally in a pAKT assay. 
     
     
         23 . The method of  claim 18  wherein the antibody fragment comprises three heavy chain CDRs set out in SEQ ID NOs: 5-7, 8-10 or 50-112 and three light chain CDRs set out in SEQ ID NOs: 11-13, 14-16 or 20-28. 
     
     
         24 . (canceled) 
     
     
         25 . The method of  claim 18 , wherein the antibody fragment comprises a heavy chain variable region selected from the group consisting of SEQ ID NO: 1 or 2 and a light chain variable region selected from the group consisting of SEQ ID NO: 3 or 4. 
     
     
         26 . The method of  claim 18  wherein the antibody fragment comprises three heavy chain CDRs set out in SEQ ID NO: 8-10 and three light chain CDRs set out in SEQ ID NO: 14-16. 
     
     
         27 . The method of  claim 18 , wherein the antibody fragment comprises a variable heavy chain amino acid sequence set out in SEQ ID NO: 2 and variable light chain amino acid sequence set out in SEQ ID NO: 4. 
     
     
         28 . (canceled) 
     
     
         29 . The method of  claim 18 , wherein the antibody fragment is from a human antibody. 
     
     
         30 . The method of  claim 18 , wherein the antibody fragment is a Fab fragment. 
     
     
         31 . (canceled) 
     
     
         32 . The method of  claim 18 , wherein the antibody fragment is in a pharmaceutical composition. 
     
     
         33 . The method of  claim 18 , wherein the antibody fragment reduces hyperinsulinemia or excess insulin signaling in the subject. 
     
     
         34 . The method of  claim 18 , wherein the subject has a blood glucose level of less than 70 mg/dL prior to administration. 
     
     
         35 . The method of  claim 18 , wherein the antibody fragment is administered at a dose of from about 0.1 to 25 mg/kg, from about 0.05 to 10 mg/kg, from about 0.3 to 6.0 mg/kg, or from about 0.1 to 3 mg/kg. 
     
     
         36 . (canceled) 
     
     
         37 . The method of  claim 18 , wherein the antibody fragment is administered in a single bolus, once every 12 hours or once per day, until hypoglycemia is eliminated. 
     
     
         38 . The method of  claim 18 , wherein the antibody is administered intravenously, intraarterially, intraperitoneally, intramuscularly, intradermally, subcutaneously or orally. 
     
     
         39 . The method of  claim 18 , wherein administration increases blood glucose in the subject by 1.5 to 10 fold or by 10 to 40%, optionally wherein administration increases blood glucose in the subject by at least 10 mg/dL. 
     
     
         40 . (canceled) 
     
     
         41 . The method of  claim 18 , wherein the subject is also on a restricted diet regimen. 
     
     
         42 . The method of  claim 18 , further comprising administering a second agent, optionally wherein the second agent is glucagon and/or insulin. 
     
     
         43 . The method of  claim 18 , wherein the administration prevents or ameliorates one or more symptoms of hypoglycemia selected from the group consisting of pancreatic nesidioblastosis, islet cell enlargement, islet cell hyperplasia, β cell budding, tachycardia, diaphoresis, flushing and reduced cognitive function. 
     
     
         44 . The method of  claim 18 , wherein the administration reduces or eliminates hypoglycemia within 20 minutes, optionally within 15 minutes. 
     
     
         45 . The method of  claim 18 , wherein the antibody fragment has a duration of action of approximately 4 hours. 
     
     
         46 . The method of  claim 18 , wherein the subject is non-responsive to dextrose or glucagon therapy.