Human EPO Mimetic Hinge Core Mimetibodies, Compositions, Methods and Uses

Abstract

The present invention relates to at least one novel human EPO mimetic hinge core mimetibody or specified portion or variant, including isolated nucleic acids that encode at least one EPO mimetic hinge core mimetibody or specified portion or variant, EPO mimetic hinge core mimetibody or specified portion or variants, vectors, host cells, transgenic animals or plants, and methods of making and using thereof, including therapeutic compositions, methods and devices.

1 . A nucleic acid encoding an EPO mimetic hinge core mimetibody comprising a polynucleotide encoding a sequence selected from the group consisting of SEQ ID NOS:82-87 and 89. 
     
     
         2 . A nucleic acid complement to the nucleic acid of  claim 1 . 
     
     
         3 . A nucleic acid encoding an EPO mimetic hinge core mimetibody comprising a polynucleotide encoding an amino acid sequence selected from the group consisting of SEQ ID NOS:1-30, or a polynucleotide complementary thereto. 
     
     
         4 . A nucleic acid encoding an EPO mimetic hinge core mimetibody comprising a polynucleotide encoding a polypeptide according to Formula (I):
   ((V( m )-P( n )-L( o )-H( p )-CH2( q )-CH3( r ))( s ),   
       where V is a portion of the N-terminus of an immunoglobulin variable region, P is a bioactive EPO mimetic peptide, L is a linker sequence, H is a portion of an immunoglobulin variable hinge core region, CH2 is a portion of an immunoglobulin CH2 constant region, CH3 is a portion of an immunoglobulin CH3 constant region, m, n, o, p, q, r, and s can independently be independently an integer between 0, 1 or 2 and 10. 
     
     
         5 . An EPO mimetic hinge core mimetibody polypeptide, comprising a sequence selected from the group consisting of SEQ ID NOS:82 and 84. 
     
     
         6 . (canceled) 
     
     
         7 . An EPO mimetic hinge core mimetibody polypeptide, comprising a sequence selected from the group consisting of SEQ ID NOS:1-30. 
     
     
         8 . An EPO mimetic hinge core mimetibody polypeptide, comprising a polypeptide according to Formula (I):
   ((V( m )-P( n )-L( o )-H( p )-CH2( q )-CH3( r ))( s ),   
       where V is QIQ, P is a bioactive peptide selected from the group consisting of SEQ ID NOS:1-30, L is selected from the group consisting of GS, GGGS (SEQ ID NO:73) and GSGGGS (SEQ ID NO:74), H is CPPCP (SEQ ID NO:75), CH2 is APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVH NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS KAK (SEQ ID NO:76), CH3 is GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGK (SEQ ID NO:78), and m, n, o, p, q, r, s are independently an integer between 0, 1 or 2 and 10. 
     
     
         9 . An EPO mimetic hinge core mimetibody polypeptide, comprising a polypeptide according to Formula (I):
   ((V( m )-P( n )-L( o )-H( p )-CH2( q )-CH3( r ))( s ),   
       where V is QIQ, P is a bioactive peptide selected from the group consisting of SEQ ID NOS:1-30, L is selected from the group consisting of GS, GGGS (SEQ ID NO:73) and GSGGGS (SEQ ID NO:74), H is CPPCP (SEQ ID NO:75), CH2 is APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVH NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS KAK (SEQ ID NO:77), CH3 is GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGK (SEQ ID NO:78), and m, n, o, p, q, r, s are independently an integer between 0, 1 or 2 and 10. 
     
     
         10 . An EPO mimetic hinge core mimetibody polypeptide, comprising a polypeptide according to Formula (I):
   ((V( m )-P( n )-L( o )-H( p )-CH2( q )-CH3( r ))( s ),   
       where V is QIQ, P is a bioactive peptide selected from the group consisting of SEQ ID NOS:1-30, L is selected from the group consisting of GS, GGGS (SEQ ID NO:73) and GSGGGS (SEQ ID NO:74), H is CPPCP (SEQ ID NO:75), CH2 is APEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT ISKAK (SEQ ID NO:79), CH3 is GQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLS LGK (SEQ ID NO:81), and m, n, o, p, q, r, s are independently an integer between 0, 1 or 2 and 10. 
     
     
         11 . An EPO mimetic hinge core mimetibody polypeptide, comprising a polypeptide according to Formula (I):
   ((V( m )-P( n )-L( o )-H( p )-CH2( q )-CH3( r ))( s ),   
       where V is QIQ, P is a bioactive peptide selected from the group consisting of SEQ ID NOS:1-30, L is selected from the group consisting of GS, GGGS (SEQ ID NO:73) and GSGGGS (SEQ ID NO:74), H is CPPCP (SEQ ID NO:75), CH2 is APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGV EVH NAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTIS KAK (SEQ ID NO:80), CH3 is GQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLS LGK (SEQ ID NO:81), and m, n, o, p, q, r, s are independently an integer between 0, 1 or 2 and 10. 
     
     
         12 . An EPO mimetic hinge core mimetibody polypeptide, comprising a polypeptide according to Formula (I):
   ((V( m )-P( n )-L( o )-H( p )-CH2( q )-CH3( r ))( s ),   
       where V is the N-terminal portion of a human variable region, P is a bioactive peptide selected from the group consisting of SEQ ID NOS:1-30, L is linker polypeptide, H is a portion of an immunoglobulin variable hinge core region, CH2 is APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVH NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS KAK (SEQ ID NO:76), CH3 is GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT QKSLSLSPGK (SEQ ID NO:78), and m, n, o, p, q, r, s are independently an integer between 0, 1 or 2 and 10. 
     
     
         13 . An EPO mimetic hinge core mimetibody polypeptide, comprising a polypeptide according to Formula (I):
   ((V( m )-P( n )-L( o )-H( p )-CH2( q )-CH3( r ))( s ),   
       where V is the N-terminal portion of a human variable region, P is a bioactive peptide selected from the group consisting of SEQ ID NOS:1-30, L is a linker polypeptide, H is a portion of an immunoglobulin variable hinge core region, CH2 is APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVH NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS KAK (SEQ ID NO:77), CH3 is GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT QKSLSLSPGK (SEQ ID NO:78), and m, n, o, p, q, r, s are independently an integer between 0, 1 or 2 and 10. 
     
     
         14 . An EPO mimetic hinge core mimetibody polypeptide, comprising a polypeptide according to Formula (I):
   ((V( m )-P( n )-L( o )-H( p )-CH2( q )-CH3( r ))( s ),   
       where V is the N-terminal portion of a human variable region, P is a bioactive peptide selected from the group consisting of SEQ ID NOS:1-30, L is a linker polypeptide, H is a portion of an immunoglobulin variable hinge core region, CH2 is APEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVE VHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT ISKAK (SEQ ID NO:79), CH3 is GQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLS LGK (SEQ ID NO:81), and m, n, o, p, q, r, s are independently an integer between 0, 1 or 2 and 10. 
     
     
         15 . An EPO mimetic hinge core mimetibody polypeptide, comprising a polypeptide according to Formula (I):
   ((V( m )-P( n )-L( o )-H( p )-CH2( q )-CH3( r ))( s ),   
       where V is the N-terminal portion of a human variable region, P is a bioactive peptide selected from the group consisting of SEQ ID NOS:1-30, L is a linker polypeptide, H is a portion of an immunoglobulin variable hinge core region, CH2 is APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGV EVH NAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTIS KAK (SEQ ID NO:80), CH3 is GQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLS LGK (SEQ ID NO:81), and m, n, o, p, q, r, s are independently an integer between 0, 1 or 2 and 10. 
     
     
         16 . An EPO mimetic hinge core mimetibody polypeptide, comprising a polypeptide according to Formula (I):
   ((V( m )-P( n )-L( o )-H( p )-CH2( q )-CH3( r ))( s ),   
       where V is QIQ, P is a bioactive peptide selected from the group consisting of SEQ ID NOS:1-30, L is a linker polypeptide, H is a portion of an immunoglobulin variable hinge core region, CH2 is a portion of an immunoglobulin CH2 constant region, CH3 is a portion of an immunoglobulin CH3 constant region, and m, n, o, p, q, r, s are independently an integer between 0, 1 or 2 and 10. 
     
     
         17 . An EPO mimetic hinge core mimetibody polypeptide, comprising a polypeptide according to Formula (I):
   ((V( m )-P( n )-L( o )-H( p )-CH2( q )-CH3( r ))( s ),   
       where V is a portion of the N-terminus of an immunoglobulin variable region, P is a bioactive EPO mimetic peptide, L is selected from the group consisting of GS, GGGS (SEQ ID NO:73) and GSGGGS (SEQ ID NO:74), H is a portion of an immunoglobulin variable hinge core region, CH2 is a portion of an immunoglobulin CH2 constant region, CH3 is a portion of an immunoglobulin CH3 constant region, and m, n, o, p, q, r, s are independently an integer between 0, 1 or 2 and 10. 
     
     
         18 . An EPO mimetic hinge core mimetibody polypeptide, comprising a polypeptide according to Formula (I):
   ((V( m )-P( n )-L( o )-H( p )-CH2( q )-CH3( r ))( s ),   
       where V is a portion of the N-terminus of an immunoglobulin variable region, P is a bioactive EPO mimetic peptide, L is a linker polypeptide, H is a portion of an immunoglobulin variable hinge core region, CH2 is a portion of an immunoglobulin CH2 constant region, CH3 is a portion of an immunoglobulin CH3 constant region, and m, n, o, p, q, r, s are independently an integer between 0, 1 or 2 and 10. 
     
     
         19 . An EPO mimetic hinge core mimetibody polypeptide, comprising a polypeptide according to Formula (I):
   ((V( m )-P( n )-L( o )-H( p )-CH2( q )-CH3( r ))( s ),   
       where V is a portion of the N-terminus of an immunoglobulin variable region, P is a bioactive EPO mimetic peptide, L is a linker polypeptide, H is CPPCP (SEQ ID NO:75), CH2 is a portion of an immunoglobulin CH2 constant region, CH3 is a portion of an immunoglobulin CH3 constant region, and m, n, o, p, q, r, s are independently an integer between 0, 1 or 2 and 10. 
     
     
         20 . An EPO mimetic hinge core mimetibody polypeptide, comprising a polypeptide according to Formula (I):
   ((V( m )-P( n )-L( o )-H( p )-CH2( q )-CH3( r ))( s ),   
       where V is a portion of the N-terminus of an immunoglobulin variable region, P is a bioactive peptide selected from the group consisting of SEQ ID NOS:1-30, L is a linker polypeptide, H is a portion of an immunoglobulin variable hinge core region, CH2 is a portion of an immunoglobulin CH2 constant region, CH3 is a portion of an immunoglobulin CH3 constant region, and m, n, o, p, q, r, s are independently an integer between 0, 1 or 2 and 10. 
     
     
         21 - 40 . (canceled) 
     
     
         41 . A method for treating a disorder associated with erythropoietin receptor activity comprising administering an effective amount of the EPO mimetic hinge core mimetibody polypeptide of  claim 7  to a patient in need thereof. 
     
     
         42 . The method of  claim 41  wherein said disorder is selected from the group consisting of, bone and joint disorders, cardiovascular disorders, a dental or oral disorder, a dermatologic disorder, an ear, nose or throat disorder, an endocrine or metabolic disorder, a gastrointestinal disorder, a gynecologic disorder, a hepatic or biliary disorder, an obstetric disorder, a hematologic disorder, an immunologic or allergic disorder, an infectious disease disorder, a musculoskeletal disorder, an oncologic disorder, a neurologic disorder, a nutritional disorder, an ophthalmologic disorder, a pediatric disorder, a poisoning disorder, a psychiatric disorder, a renal disorder, and a pulmonary disorder. 
     
     
         43 . The method of  claim 42  wherein said hematologic disorder is selected from the group consisting of cancer treatment related anemia, radiotherapy or chemotherapy related anemia, viral or bacterial infection treatment related anemia, renal anemia, anemia of prematurity, pediatric and/or adult cancer-associated anemia, anemia associated with lymphoma, myeloma, multiple myeloma, AIDS-associated anemia, cyclic neutropenia or Kostmann syndrome (congenital agranulocytosis), end-stage renal disease, anemia associated with dialysis, chronic renal insufficiency, congenital hypoplastic anemia, thalassemia major, sickle cell disease, and vaso-occlusive complications of sickle cell disease.