Extended recombinant polypeptides and compositions comprising same
The present invention relates to compositions comprising biologically active proteins linked to extended recombinant polypeptide (XTEN), isolated nucleic acids encoding the compositions and vectors and host cells containing the same, and methods of using such compositions in treatment of glucose-related diseases, metabolic diseases, coagulation disorders, and growth hormone-related disorders and conditions.
1. A method of treating a disease, disorder or condition, comprising administering a therapeutically effective dose of a pharmaceutical composition to a subject in need thereof,
wherein the pharmaceutical composition comprises a fusion protein and one or more pharmaceutically acceptable excipients,
wherein the fusion protein comprises an extended recombinant polypeptide linked to a biologically active protein (BP),
wherein the extended recombinant polypeptide comprises an amino acid sequence which has at least 90% sequence identity to SEQ ID NOs: 205, 208, 211, 461-465, 467-477, 479, 480-485, 487-489, 491-504, 699, 772, or 793,
wherein the extended recombinant polypeptide comprises a motif of SEQ ID NO: 190, SEQ ID NO: 192, or SEQ ID NO: 193,
wherein the disease, disorder or condition is type 1 diabetes, type 2 diabetes, obesity, hyperglycemia, hyperinsulinemia, decreased insulin production, insulin resistance, syndrome X, excessive appetite, insufficient satiety, glucagonomas, dyslipidemia, retinal neurodegenerative processes, Factor VII deficiency, Factor X deficiency, Factor XII deficiency, hemophilia A, hemophilia B, Von Willebrand's disease, hypertension, acute coronary syndrome, rheumatoid arthritis, reperfusion injury following ischemia, growth-hormone deficiency, Turner's Syndrome, Prader-Willi Syndrome, idiopathic short stature, AIDS wasting, multiple sclerosis, Crohn's disease, ulcerative colitis, muscular dystrophy, or surgical bleeding.
2. The method of claim 1 , wherein the pharmaceutical composition is administered subcutaneously, intramuscularly, or intravenously.
3. The method of claim 1 , wherein the extended recombinant polypeptide is further characterized in that:
(a) the sum of asparagine and glutamine residues is less than 10% of the total amino acid sequence of the extended recombinant polypeptide; and/or
(b) the sum of methionine and tryptophan residues is less than 2% of the total amino acid sequence of the extended recombinant polypeptide.
4. The method of claim 1 , wherein the extended recombinant polypeptide is further characterized in that:
(a) no one type of amino acid constitutes more than 30% of the extended recombinant polypeptide sequence;
(b) the extended recombinant polypeptide comprises a sequence in which no three contiguous amino acids are identical unless the amino acid is serine, in which case no more than three contiguous amino acids are serine residues; and/or
(c) the extended recombinant polypeptide sequence has a subsequence score of less than 10.
5. The method of claim 1 wherein the fusion protein further comprises a second extended recombinant polypeptide sequence.
6. The method of claim 1 , wherein the fusion protein further comprises a spacer sequence between the BP and the extended recombinant polypeptide, wherein the spacer sequence comprises a cleavage sequence.
7. The method of claim 6 , wherein the cleavage sequence is susceptible to cleavage by FXIa, FXIIa, kallikrein, FVIIa, FIXa, FXa, thrombin, elastase-2, granzyme B, MMP-12, MMP-13, MMP-17 or MMP-20, TEV, enterokinase, rhinovirus 3C protease, or sortase A.