Polypeptide-polynucleotide-complex and its use in targeted effector moiety delivery

Abstract

Herein is reported a polypeptide-polynucleotide-complex as therapeutic agent and its use as tool for the targeted delivery of an effector moiety. The polynucleotide part of the complex is essentially resistant to proteolytic and enzymatic degradation in vivo. Additionally the polypeptide part specifically binds to a compound or structure such as a tissue or organ, a process or a disease. Thus, one aspect as reported herein is a polypeptide-polynucleotide-complex comprising a) a polypeptide specifically binding to a target and conjugated to a first member of a binding pair, b) a polynucleotide linker conjugated at its first terminus to the second member of the binding pair, and c) an effector moiety conjugated to a polynucleotide that is complementary to at least a part of the polynucleotide linker.

1 . A complex comprising
 a) a first polypeptide that specifically binds to a first target and that is conjugated to a first member of a first binding pair,   b) a second polypeptide that specifically binds to a second target and that is conjugated to a first member of a second binding pair, and   c) a ss-L-DNA-linker conjugated to the second member of the first binding pair and conjugated to the second member of the second binding pair.   
     
     
         2 . A complex comprising
 a) a polypeptide that specifically binds to a target and that is conjugated to a first member of a binding pair,   b) a ss-L-DNA-linker conjugated to the second member of the binding pair, and   c) an effector moiety conjugated to a polynucleotide that is complementary to at least a part of the ss-L-DNA-linker.   
     
     
         3 . The complex according to  claim 1 , further comprising an effector moiety conjugated to a polynucleotide that is complementary to at least a part of the ss-L-DNA-linker. 
     
     
         4 . The complex according to  claim 1 , further comprising an effector moiety conjugated to a polynucleotide (L-DNA) that is complementary to at least a part of the ss-L-DNA-linker. 
     
     
         5 . The complex according to  claim 1 , characterized in that the first polypeptide is a monovalent antibody or monovalent antibody fragment. 
     
     
         6 . The complex according to  claim 5 , characterized in that the first and second polypeptide bind to the same target and to non-overlapping epitopes on the same target. 
     
     
         7 . The complex according to  claim 1 , characterized in that the members of the binding pairs are selected from the group consisting of leucine zipper domain dimers and hybridizing nucleic acid sequences. 
     
     
         8 . The complex according to  claim 1 , characterized in that the complex is a non-covalent complex. 
     
     
         9 . The complex according to  claim 1 , characterized in that the effector moiety is selected from the group consisting of a binding moiety, a labeling moiety and a biologically active moiety. 
     
     
         10 . The complex according to  claim 1 , characterized in that the first polypeptide is a FAB′ fragment of the anti-HER2 antibody 2C4, the second polypeptide is a FAB′ fragment of the anti-HER2 antibody 4D5, the members of the binding pair are hybridizing nucleic acids and the ss-L-DNA-linker comprises 60 to 100 L-DNA nucleotides. 
     
     
         11 . The complex according to  claim 1 , characterized in that the first and second member of the first binding pair comprise the nucleic acid sequences of SEQ ID NO: 05 and SEQ ID NO: 08. 
     
     
         12 . The complex according to  claim 1 , characterized in that the first and second member of the second binding pair comprise the nucleic acid sequences of SEQ ID NO: 06 and SEQ ID NO: 07. 
     
     
         13 . A method of producing a complex comprising the components
 a) a polypeptide that specifically binds to a target and that is conjugated to a first member of a binding pair,   b) a polynucleotide linker conjugated to the second member of the binding pair, and   c) an effector moiety conjugated to a polynucleotide that is complementary to at least a part of the polynucleotide linker,   comprising the steps of: a) synthesizing the polypeptide specifically binding to a target and conjugated to a first member of a binding pair, and synthesizing an effector moiety conjugated to a polynucleotide that is complementary to at least a part of the polynucleotide linker, respectively, b) synthesizing the polynucleotide linker conjugated at its first terminus to the second member of the binding pair, and c) forming the polypeptide-polynucleotide-complex by hybridizing the synthesized components.   
     
     
         14 . A method of producing a complex comprising the components
 a) a first polypeptide that specifically binds to a first target and that is conjugated to a first member of a first binding pair,   b) a second polypeptide that specifically binds to a second target and that is conjugated to a first member of a second binding pair, and   c) a polynucleotide linker conjugated to the second member of the first binding pair and conjugated to the second member of the second binding pair,   comprising the steps of: a) synthesizing the first polypeptide specifically binding to a first target which is conjugated to a first member of a first binding pair, and synthesizing the second polypeptide specifically binding to a second target which is conjugated to a first member of a second binding pair, respectively, and b) synthesizing the polynucleotide linker conjugated at its first terminus to the second member of the first binding pair and conjugated at its second terminus to the second member of the second binding pair, and c) forming the polypeptide-polynucleotide-complex by hybridizing the synthesized components.   
     
     
         15 . A pharmaceutical formulation comprising the complex according to  claim 1  and optionally a pharmaceutically acceptable carrier. 
     
     
         16 . The complex according to  claim 1  for use as a medicament. 
     
     
         17 . Use of the complex according to  claim 1  in the manufacture of a medicament.