Follicle stimulating hormone superagonists

Abstract

Modified follicle stimulating hormone (FSH), containing an amino acid sequence which differs from the wild-type FSH, said modified FSH comprising a modified α-subunit, wherein the potency of said modified FSH is increased by at least about a ten fold as compared to wild type FSH. Nucleic acids encoding the modified FSH, vectors comprising the nucleic acids, and host cells comprising the vectors. Methods of assisting reproduction, diagnosing and/or treating conditions associated with glycoprotein hormone activity, reducing hyperstimulation syndrome, improving the quality of oocytes, inducing superovulation, and enhancing in vitro fertilization.

1 . A modified follicle stimulating hormone (FSH), containing an amino acid sequence which differs from the wild-type FSH, said modified FSH comprising a modified α-subunit, wherein the potency of said modified FSH is increased by at least about a ten fold as compared to wild type FSH. 
     
     
         2 .- 39 . (canceled) 
     
     
         40 . The modified FSH of  claim 1 , wherein said modified FSH is human, wherein said α-subunit comprises at least two basic amino acids at positions corresponding to positions 13, 14, 16, 17, 20, 21, 22, 66, 68, 73, 74, and 81 of SEQ ID NO:1, and wherein said basic amino acids are selected from the group consisting of lysine and arginine. 
     
     
         41 . The modified FSH of  claim 1 , further comprising a modified β-subunit. 
     
     
         42 .- 43 . (canceled) 
     
     
         44 . A nucleic acid encoding the modified FSH α-subunit of  claim 1 . 
     
     
         45 . A vector comprising a nucleic acid encoding the modified FSH α-subunit of  claim 1 , wherein the vector is suitable for expressing the nucleic acid. 
     
     
         46 . A host cell comprising the vector of  claim 45 , wherein the host cell is suitable for expressing the nucleic acid. 
     
     
         47 .- 51 . (canceled) 
     
     
         52 . The modified FSH of  claim 1 , wherein the plasma half-life is increased as compared to wild type FSH. 
     
     
         53 . The modified FSH of  claim 52 , wherein said modified FSH further comprises at least one sequence with a potential glycosylation site selected from the group consisting of a sequence comprising a N-glycosylation site and a sequence comprising an O-glycosylation site. 
     
     
         54 . The modified FSH of  claim 53 , wherein said at least one sequence with a potential glycosylation recognition site is an N-terminal extension on said α chain. 
     
     
         55 . The modified FSH of  claim 54 , wherein said N-terminal extension is selected from the group consisting of ANITV (SEQ ID No. 3) and ANITVNITV (SEQ ID No. 4). 
     
     
         56 .- 58 . (canceled) 
     
     
         59 . The modified FSH of  claim 52 , wherein said modified FSH is altered to increase the number of negatively charged residues within the molecule to increase plasma half-life. 
     
     
         60 . The method of  claim 59 , wherein said negatively charged residues are selected from the group consisting of glutamate aspartate. 
     
     
         61 .- 66 . (canceled) 
     
     
         67 . A method of assisting reproduction in a subject comprising administering an assisting amount of the modified FSH of  claim 1 . 
     
     
         68 . A method of diagnosing and/or treating a condition associated with a glycoprotein hormone activity in a patient comprising administering an effective amount of the modified FSH of  claim 1  to the patient. 
     
     
         69 .- 82 . (canceled) 
     
     
         83 . A method of reducing hyperstimulation syndrome in a patient undergoing assisted reproduction therapy, comprising
 (a) administering an assisting amount of a first modified FSH according to  claim 1  wherein the plasma half-life of said first modified FSH is increased as compared to wild type FSH, and   (b) subsequently administering an assisting amount of a second modified FSH according to  claim 1  wherein the plasma half-life of said second modified FSH is decreased as compared to said first modified FSH, wherein ovarian hyperstimulation is reduced as compared to when the same patient is administered only said first modified FSH during assisted reproduction therapy.   
     
     
         84 . A method of improving the quality of oocytes in an animal comprising:
 administering an effective amount of a superactive follicle stimulating hormone to said animal, wherein said superactive follicle stimulating hormone contains an D-subunit with a basic amino acid at one or more positions selected from the group consisting of positions 13, 14, 16, and 20.   
     
     
         85 .- 103 . (canceled) 
     
     
         104 . A method of inducing superovulation in an animal comprising:
 administering an effective amount of superactive follicle stimulating hormone to said animal,   wherein said superactive follicle stimulating hormone contains an alpha-subunit with a basic amino acid at one or more positions selected from the group consisting of positions 13, 14, 16, and 20. 105. A method of  claim 104 , wherein superovulation is characterized by an increase in oocyte number as compared to a like animal receiving the same amount of recombinant wild type FSH.   
     
     
         105 .- 119 . (canceled) 
     
     
         120 . A method of enhancing in vitro fertilization comprising:
 administering an effective amount of superactive follicle stimulating hormone to said animal,   wherein said superactive follicle stimulating hormone contains an alpha-subunit with a basic amino acid at one or more positions selected from the group consisting of positions 13, 14, 16, and 20.   
     
     
         120 .- 137 . (canceled)