Methods for the production of functional protein from dna having a nonsense mutation and the treatment of disorders associated therewith

Abstract

The present invention relates to functional proteins encoded by nucleic acid sequences comprising a nonsense mutation. The present invention also relates to methods for the production of functional proteins encoded by nucleic acid sequences comprising a nonsense mutation and the use of such proteins for prevention, management and/or treatment of diseases associated with a nonsense mutation(s) in a gene.

1 - 20 . (canceled) 
     
     
         21 . A method for the treatment of cystic fibrosis in a human patient with a nonsense mutation at one or more positions in the CFTR gene, comprising administering to the patient having cystic fibrosis and a nonsense mutation at one or more positions in the CFTR gene, an effective amount of 3-[5-(2-fluoro-phenyl)-[1,2,4]oxadiazol-3-yl]-benzoic acid or a pharmaceutically acceptable salt thereof, in three doses, wherein the three doses comprise a first dose, a second dose and a third dose, and wherein the amounts of the first dose and the second dose are the same and the amount of the third dose is twice the amount of the first dose, in a plurality of 24 hour time periods, wherein the second dose is administered about 6 hours after the first dose is administered, the third dose is administered about 6 hours after the second dose is administered, and the first dose for a next 24 hour time period is administered about 12 hours after the third dose was administered for a preceding 24 hour time period, and wherein a plasma concentration of 3-[5-(2-fluoro-phenyl)-[1,2,4]oxadiazol-3-yl]-benzoic acid or a pharmaceutically acceptable salt thereof in a range of about 2 μg/mL to about 20 μg/mL is maintained in said patient for a 24 hour time period, wherein said patient has been determined to have a likelihood of responding to treatment through a pre-treatment patient screening, said patient screening comprising contacting a cell sample from said patient in vitro with 3-[5-(2-fluoro-phenyl)-[1,2,4]oxadiazol-3-yl]-benzoic acid or a pharmaceutically acceptable salt thereof, and measuring the expression or activity of CFTR protein produced in the presence of 3-[5-(2-fluoro-phenyl)-[1,2,4]oxadiazol-3-yl]-benzoic acid or a pharmaceutically acceptable salt thereof compared to the absence of 3-[5-(2-fluoro-phenyl)-[1,2,4]oxadiazol-3-yl]-benzoic acid or a pharmaceutically acceptable salt thereof, wherein increased expression or activity of functional readthrough CFTR protein in said contacted cell sample relative to a cell sample from said patient not contacted with 3-[5-(2-fluoro-phenyl)-[1,2,4]oxadiazol-3-yl]-benzoic acid or a pharmaceutically acceptable salt thereof indicates a likelihood that said patient will respond to treatment. 
     
     
         22 . The method of  claim 21 , wherein the cell sample is from the nose of a patient undergoing pre-treatment screening. 
     
     
         23 . The method of  claim 21 , wherein the nonsense mutation at one or more positions in the CFTR gene is found at least at positions selected from 414, 1609, 3976 and 1282 of the CFTR gene. 
     
     
         24 . The method of  claim 23 , wherein the cell sample is from the nose of a patient undergoing pre-treatment screening.