Therapeutic agents comprising elastin-like peptides

Abstract

The present invention provides therapeutic agents and compositions comprising elastin-like peptides (ELPs) and therapeutic proteins. In some embodiments, the therapeutic protein is a GLP-1 receptor agonist, insulin, or Factor VII/VIIa, including functional analogs. The present invention further provides encoding polynucleotides, as well as methods of making and using the therapeutic agents. The therapeutic agents have improvements in relation to their use as therapeutics, including, inter alia, one or more of half-life, clearance and/or persistance in the body, solubility, and bioavailability.

What is claimed is: 
     
         1 . A therapeutic agent comprising an elastin-like peptide (ELP) component and a therapeutic component, the therapeutic component having an extended circulatory half-life and/or a lower therapeutically effective dose when compared to the therapeutic component alone. 
     
     
         2 . The therapeutic agent of  claim 1 , wherein the therapeutic component is a therapeutic protein listed in Table 1 or a functional portion or functional analog thereof. 
     
     
         3 . The therapeutic agent of  claim 2 , wherein an ELP component is covalently bonded to the therapeutic protein at an N- and/or C-terminus thereof. 
     
     
         4 . The therapeutic agent of any one of  claims 1 - 3 , wherein a therapeutic component is covalently bonded to the ELP component at the N-terminus of the ELP component, and a second therapeutic component is covalently bonded to the ELP component at the C-terminus of the ELP component. 
     
     
         5 . The therapeutic agent of any one of  claims 1 - 4 , wherein a therapeutic component is covalently bonded to the ELP component at an N- and/or C-terminus thereof. 
     
     
         6 . The therapeutic agent of any one of  claims 1 - 5 , wherein a first ELP component is covalently bonded to the therapeutic component at the N-terminus of the therapeutic component, and a second ELP component is covalently bonded to the therapeutic component at the C-terminus of the therapeutic component. 
     
     
         7 . The therapeutic agent of any one of  claims 1 - 6 , further comprising a spacer moiety between the ELP component and the therapeutic component. 
     
     
         8 . The therapeutic agent of  claim 7 , wherein the spacer moiety comprises one or more of a protease-resistant moiety, a non-peptide chemical moiety, and a protease cleavage site. 
     
     
         9 . The therapeutic agent of  claim 8 , wherein the protease cleavage site is a thrombin cleavage site, a factor Xa cleavage site, a metalloprotease cleavage site, an enterokinase cleavage site, a Tev cleavage site, and a cathepsin cleavage site. 
     
     
         10 . The therapeutic agent of  claim 9 , wherein the spacer moiety comprises a non-cleavable moiety having the formula [(Gly) n -Ser] m  (SEQ ID NO: 22) where n is from 1 to 4, inclusive, and m is from 1 to 4, inclusive. 
     
     
         11 . The therapeutic agent of any one of  claims 1 - 10 , wherein the ELP comprises at least one repeating unit selected from SEQ ID NOS: 1-12. 
     
     
         12 . The therapeutic agent of  claim 11 , wherein said repeating unit is VPGXG (SEQ ID NO: 3). 
     
     
         13 . The therapeutic agent of  claim 12 , wherein X is any natural or non-natural amino acid residue, and wherein X varies among at least two units. 
     
     
         14 . The therapeutic agent of  claim 13 , wherein each X is independently selected from alanine, arginine, asparagine, aspartic acid, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, serine, threonine, tryptophan, tyrosine and valine residues. 
     
     
         15 . The therapeutic agent of any one of  claims 1 - 14 , wherein the therapeutic agent has a molecular weight of less than about 50 kDa. 
     
     
         16 . The therapeutic agent of any one of  claims 1 - 15 , wherein the ELP component has a Tt greater than 37° C. 
     
     
         17 . The therapeutic agent of any one of  claims 1 - 16 , wherein the therapeutic agent is a genetically encoded fusion protein. 
     
     
         18 . A polynucleotide comprising a nucleotide sequence encoding the therapeutic agent of  claim 17 . 
     
     
         19 . The polynucleotide of  claim 18 , further comprising an expression control element operably linked to said nucleotide sequence. 
     
     
         20 . The polynucleotide of  claim 19 , wherein said expression control element comprises a promoter. 
     
     
         21 . A vector comprising the polynucleotide of any one of  claims 18 - 20 . 
     
     
         22 . An isolated host cell containing the vector of  claim 21  or the polynucleotide of any one of  claims 18 - 20 . 
     
     
         23 . A pharmaceutical composition comprising the therapeutic agent of any one of  claims 1 - 18 , and a pharmaceutically acceptable carrier and/or excipient. 
     
     
         24 . A method of treating or preventing a disease, disorder, or condition in a subject, comprising administering an effective amount of the therapeutic agent of any one of  claims 1 - 17  or the pharmaceutical composition of  claim 23  to a subject in need thereof. 
     
     
         25 . The method of  claim 24 , wherein the subject has an indication listed in Table 1. 
     
     
         26 . A therapeutic agent comprising an elastin-like peptide (ELP) component and a glucagon-like peptide (GLP)-1 receptor agonist. 
     
     
         27 . The therapeutic agent of  claim 26 , wherein the GLP-1 receptor agonist is GLP-1 or functional analog thereof. 
     
     
         28 . The therapeutic agent of  claim 26 , wherein the GLP-1 receptor agonist is an exendin-4 or functional analog thereof. 
     
     
         29 . The therapeutic agent of any one of  claims 26 - 28 , wherein the ELP component is covalently bonded to the GLP-1 receptor agonist at an N- and/or C-terminus thereof. 
     
     
         30 . The therapeutic agent of any one of  claims 26 - 29 , wherein a first GLP-1 receptor agonist is covalently bonded to the ELP component at the N-terminus of the ELP component, and a second GLP-1 receptor agonist is covalently bonded to the ELP component at the C-terminus of the ELP component. 
     
     
         31 . The therapeutic agent of any one of  claims 26 - 30 , wherein the GLP-1 receptor agonist is covalently bonded to the ELP component at an N- and/or C-terminus thereof. 
     
     
         32 . The therapeutic agent of any one of  claims 26 - 31 , wherein a first ELP component is covalently bonded to the GLP-1 receptor agonist at the N-terminus of the GLP-1 receptor agonist, and a second ELP component is covalently bonded to the GLP-1 receptor agonist at the C-terminus of the GLP-1 receptor agonist. 
     
     
         33 . The therapeutic agent of any one of  claims 26 - 32 , further comprising a spacer moiety between the ELP component and GLP-receptor agonist. 
     
     
         34 . The therapeutic agent of  claim 33 , wherein the spacer moiety comprises one or more of a protease-resistant moiety, a non-peptide chemical moiety, and a protease cleavage site. 
     
     
         35 . The therapeutic agent of  claim 34 , wherein the protease cleavage site is a thrombin cleavage site, a factor Xa cleavage site, a metalloprotease cleavage site, an enterokinase cleavage site, a Tev cleavage site, and a cathepsin cleavage site. 
     
     
         36 . The therapeutic agent of  claim 34 , wherein the spacer moiety comprises a non-cleavable moiety having the formula [(Gly) n -Ser] m  (SEQ ID NO.: 22) where n is from 1 to 4, inclusive, and m is from 1 to 4, inclusive. 
     
     
         37 . The therapeutic agent of any one of  claims 26 - 36 , wherein the ELP comprises at least one repeating unit selected from SEQ ID NOS: 1-12. 
     
     
         38 . The therapeutic agent of  claim 37 , wherein said repeating unit is VPGXG (SEQ ID NO: 3). 
     
     
         39 . The therapeutic agent of  claim 38 , wherein X is any natural or non-natural amino acid residue, and wherein X varies among at least two units. 
     
     
         40 . The therapeutic agent of  claim 38 , wherein each X is independently selected from alanine, arginine, asparagine, aspartic acid, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, serine, threonine, tryptophan, tyrosine and valine residues. 
     
     
         41 . The therapeutic agent of any one of  claims 26 - 40 , wherein the therapeutic agent has a molecular weight of less than about 50 kDa. 
     
     
         42 . The therapeutic agent of any one of  claims 26 - 40 , wherein the ELP component has a Tt greater than 37° C. 
     
     
         43 . The therapeutic agent of any one of  claims 26 - 42 , wherein the therapeutic agent is a genetically encoded fusion protein. 
     
     
         44 . A polynucleotide comprising a nucleotide sequence encoding the therapeutic agent of  claim 43 . 
     
     
         45 . The polynucleotide of  claim 44 , further comprising an expression control element operably linked to said nucleotide sequence. 
     
     
         46 . The polynucleotide of  claim 45 , wherein said expression control element comprises a promoter. 
     
     
         47 . A vector comprising the polynucleotide of any one of  claims 44 - 46 . 
     
     
         48 . An isolated host cell containing the vector of  claim 47  or the polynucleotide of any one of  claims 44 - 46 . 
     
     
         49 . A pharmaceutical composition comprising the therapeutic agent of any one of  claims 26 - 43 , and a pharmaceutically acceptable carrier and/or excipient. 
     
     
         50 . A method of treating or preventing a biological condition, disorder, or disease in a subject, comprising administering an effective amount of the therapeutic agent of any one of  claims 26 - 43  or the pharmaceutical composition of  claim 49  to a subject in need thereof. 
     
     
         51 . The method of  claim 50 , wherein the subject has diabetes. 
     
     
         52 . A therapeutic agent comprising an elastin-like peptide (ELP) component and an insulin or functional analog thereof. 
     
     
         53 . The therapeutic agent of  claim 52 , wherein the ELP component is covalently bonded to the insulin or functional analog at an N- and/or C-terminus thereof. 
     
     
         54 . The therapeutic agent of  claim 52  or  53 , wherein a first insulin or functional analog is covalently bonded to the ELP component at the N-terminus of the ELP component, and a second insulin or functional analog is covalently bonded to the ELP component at the C-terminus of the ELP component. 
     
     
         55 . The therapeutic agent of  claim 52 , wherein the insulin or functional analog is covalently bonded to the ELP component at an N- and/or C-terminus thereof. 
     
     
         56 . The therapeutic agent of  claim 52  or  53 , wherein a first ELP component is covalently bonded to the insulin or functional analog at an N-terminus thereof, and a second ELP component is covalently bonded to the insulin or functional analog at a C-terminus thereof. 
     
     
         57 . The therapeutic agent of any one of  claims 52 - 56 , further comprising a spacer moiety between the ELP component and the insulin or functional analog. 
     
     
         58 . The therapeutic agent of  claim 57 , wherein the spacer moiety comprises one or more of a protease-resistant moiety, a non-peptide chemical moiety, and a protease cleavage site. 
     
     
         59 . The therapeutic agent of  claim 58 , wherein the protease cleavage site is a thrombin cleavage site, a factor Xa cleavage site, a metalloprotease cleavage site, an enterokinase cleavage site, a Tev cleavage site, or a cathepsin cleavage site. 
     
     
         60 . The therapeutic agent of  claim 58 , wherein the spacer moiety comprises a non-cleavable moiety having the formula [(Gly) n -Ser] m  (SEQ ID NO: 22) where n is from 1 to 4, inclusive, and m is from 1 to 4, inclusive. 
     
     
         61 . The therapeutic agent of any one of  claims 52 - 60 , wherein the ELP component comprises at least one repeating unit selected from SEQ ID NOS: 1-12. 
     
     
         62 . The therapeutic agent of  claim 61 , wherein said repeating unit is VGPXG (SEQ ID NO: 3). 
     
     
         63 . The therapeutic agent of  claim 61 , wherein the repeating unit is Ile-Pro-Gly-X-Gly (SEQ ID NO: 5) or Leu-Pro-Gly-X-Gly (SEQ ID NO: 7), wherein X is any natural or non-natural amino acid residue, and wherein X varies among at least two units. 
     
     
         64 . The therapeutic agent of  claim 62  or  63 , wherein each X is independently selected from alanine, arginine, asparagine, aspartic acid, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, serine, threonine, tryptophan, tyrosine and valine residues. 
     
     
         65 . The therapeutic agent of any one of  claims 52 - 64 , wherein the therapeutic agent has a molecular weight of less than about 50 kDa. 
     
     
         66 . The therapeutic agent of any one of  claims 52 - 65 , wherein the ELP component has a Tt greater than 37° C. 
     
     
         67 . The therapeutic agent of any one of  claims 52 - 66 , wherein the therapeutic agent is a genetically encoded fusion protein. 
     
     
         68 . A polynucleotide comprising a nucleotide sequence encoding the therapeutic agent of  claim 67 . 
     
     
         69 . The polynucleotide of  claim 68 , further comprising an expression control element operably linked to said nucleotide sequence. 
     
     
         70 . The polynucleotide of  claim 69 , wherein said expression control element comprises a promoter. 
     
     
         71 . A vector comprising the polynucleotide of any one of  claims 68 - 70 . 
     
     
         72 . An isolated host cell containing the vector of  claim 71  or the polynucleotide of any one of  claims 68 - 70 . 
     
     
         73 . A pharmaceutical composition comprising the therapeutic agent of any one of  claims 52 - 67 , and a pharmaceutically acceptable carrier and/or excipient. 
     
     
         74 . A method of preventing or treating a biological condition, disorder, or disease in a subject, comprising administering an effective amount of the therapeutic agent of any one of  claims 52 - 67  or the pharmaceutical composition of  claim 73  to a subject in need thereof. 
     
     
         75 . The method of  claim 74 , wherein the subject has diabetes. 
     
     
         76 . A therapeutic agent comprising an elastin-like peptide (ELP) component and a Factor VII/VIIa or functional analog thereof. 
     
     
         77 . The therapeutic agent of  claim 76 , wherein the ELP component is covalently bonded to the Factor VII/VIIa or functional analog at an N- and/or C-terminus of the Factor VII/VIIa. 
     
     
         78 . The therapeutic agent of  claim 76  or  77 , wherein a first Factor VII/VIIa or functional analog is covalently bonded to the ELP at the N-terminus of the ELP component, and a second Factor VII/VIIa or functional analog is covalently bonded to the ELP component at the C-terminus of the ELP component. 
     
     
         79 . The therapeutic agent of  claim 76 , wherein the Factor VII/VIIa or functional analog is covalently bonded to the ELP component at an N- and/or C-terminus of the ELP component. 
     
     
         80 . The therapeutic agent of  claim 76  or  77 , wherein a first ELP component is covalently bonded to the Factor VII/VIIa or functional analog at an N-terminus thereof, and a second ELP component is covalently bonded to the Factor VII/VIIa or functional analog at a C-terminus thereof. 
     
     
         81 . The therapeutic agent of any one of  claims 76 - 80 , further comprising a spacer moiety between the ELP component and factor VII/VIIa or functional analog. 
     
     
         82 . The therapeutic agent of  claim 81 , wherein the spacer moiety comprises one or more of a protease-resistant moiety, a non-peptide chemical moiety, and a protease cleavage site. 
     
     
         83 . The therapeutic agent of  claim 82 , wherein the protease cleavage site is a thrombin cleavage site, a factor Xa cleavage site, a metalloprotease cleavage site, an enterokinase cleavage site, a Tev cleavage site, and a cathepsin cleavage site. 
     
     
         84 . The therapeutic agent of  claim 82 , wherein the spacer moiety comprises a non-cleavable moiety having the formula [(Gly) n -Ser] m  (SEQ ID NO: 22) where n is from 1 to 4, inclusive, and m is from 1 to 4, inclusive. 
     
     
         85 . The therapeutic agent of any one of  claims 76 - 84 , wherein the ELP component comprises at least one repeating unit selected from SEQ ID NOS: 1-12. 
     
     
         86 . The therapeutic agent of  claim 85 , wherein said repeating unit is VPGXG (SEQ ID NO: 3). 
     
     
         87 . The therapeutic agent of  claim 86 , wherein X is any natural or non-natural amino acid residue, and wherein X varies among at least two units. 
     
     
         88 . The therapeutic agent of  claim 86 , wherein each X is independently selected from alanine, arginine, asparagine, aspartic acid, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, serine, threonine, tryptophan, tyrosine and valine residues. 
     
     
         89 . The therapeutic agent of any one of  claims 76 - 88 , wherein the therapeutic agent has a molecular weight of less than about 70 kDa. 
     
     
         90 . The therapeutic agent of any one of  claims 76 - 89 , wherein the ELP component has a Tt greater than 37° C. 
     
     
         91 . The therapeutic agent of any one of  claims 76 - 90 , wherein the therapeutic agent is a genetically encoded fusion protein. 
     
     
         92 . A polynucleotide comprising a nucleotide sequence encoding the therapeutic agent of  claim 91 . 
     
     
         93 . The polynucleotide of  claim 92 , further comprising an expression control element operably linked to said nucleotide sequence. 
     
     
         94 . The polynucleotide of  claim 93 , wherein said expression control element comprises a promoter. 
     
     
         95 . A vector comprising the polynucleotide of any one of  claims 92 - 94 . 
     
     
         96 . An isolated host cell containing the vector of  claim 95  or the polynucleotide of any one of  claims 92 - 95 . 
     
     
         97 . A pharmaceutical composition comprising the therapeutic agent of any one of  claims 76 - 91 , and a pharmaceutically acceptable carrier and/or excipient. 
     
     
         98 . A method of preventing or treating a subject for a biological condition, disorder, or disease, comprising administering an effective amount of the therapeutic agent of any one of  claims 76 - 91  or the pharmaceutical composition of  claim 97  to a subject in need thereof. 
     
     
         99 . The method of  claim 98 , wherein the subject has a hemorrhage. 
     
     
         100 . The method of  claim 98  or  99 , wherein the subject has hemophilia.