IMMUNOGLOBULIN CHIMERIC MONOMER-DIMER HYBRIDS

The invention relates to a chimeric monomer-dimer hybrid protein wherein the protein comprises a first and a second polypeptide chain, the first polypeptide chain comprising at least a portion of an immunoglobulin constant region and a biologically active molecule, and the second polypeptide chain comprising at least a portion of an immunoglobulin constant region without the biologically active molecule of the first chain. The invention also relates to methods of using and methods of making the chimeric monomer-dimer hybrid protein of the invention. 1194.-. (canceled)195. A pharmaceutical composition comprising (a) a chimeric protein and (b) a pharmaceutically acceptable carrier , wherein the chimeric protein comprises a first and second polypeptide chain , wherein the first polypeptide chain comprises (i) a biologically active molecule selected from a hormone , a cytokine , a growth factor , and a cell surface receptor , and (ii) an immunoglobulin constant region or a portion thereof; and wherein the second polypeptide chain comprises an immunoglobulin constant region or a portion thereof without a biologically active molecule.196. The pharmaceutical composition of claim 195 , wherein one of the immunoglobulin constant regions or portions thereof is an Fc fragment.197. The pharmaceutical composition of claim 195 , wherein the immunoglobulin constant region or a portion thereof in the first polypeptide chain and the immunoglobulin constant region or a portion thereof in the second polypeptide chain are identical.198. The pharmaceutical composition of claim 195 , wherein the biologically active molecule comprises a hormone.199. The pharmaceutical composition of claim 198 , wherein the hormone is selected from human growth hormone (hGH) claim 198 , gonadotropin releasing hormone (GnRH) claim 198 , leuprolide claim 198 , follicle stimulating hormone claim 198 , progesterone claim 198 , estrogen claim 198 , and testosterone.200. The pharmaceutical composition of claim ...

IMMUNOGLOBULIN CHIMERIC MONOMER-DIMER HYBRIDS

The invention relates to a chimeric monomer-dimer hybrid protein wherein the protein comprises a first and a second polypeptide chain, the first polypeptide chain comprising at least a portion of an immunoglobulin constant region and a biologically active molecule, and the second polypeptide chain comprising at least a portion of an immunoglobulin constant region without the biologically active molecule of the first chain. The invention also relates to methods of using and methods of making the chimeric monomer-dimer hybrid protein of the invention.

OPTIMIZED FACTOR IX GENE

The present invention provides codon optimized Factor IX sequences, vectors and host cells comprising codon optimized Factor IX sequences, polypeptides encoded by codon optimized Factor IX sequences, and methods of producing such polypeptides. The present invention also provides methods of treating bleeding disorders such as hemophilia comprising administering to the subject a codon optimized Factor IX nucleic acid sequence or the polypeptide encoded thereby. 1. An isolated nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of:(i) a sequence at least 85% identical to SEQ ID NO: 1,(ii) a sequence at least 95% identical to SEQ ID NO: 54,(iii) a sequence at least 96% identical to SEQ ID NO: 55,(iv) a sequence at least 99% identical to SEQ ID NO: 56,(v) a sequence at least 94% identical to SEQ ID NO: 57, and(vi) a sequence at least 94% identical to SEQ ID NO: 58;wherein the nucleotide sequence encodes a polypeptide with Factor IX activity.214-. (canceled)15. The isolated nucleic acid molecule of claim 1 , wherein:(i) the human codon adaptation index is increased relative to SEQ ID NO:2;(ii) the nucleotide sequence contains a higher percentage of G/C nucleotides compared to the percentage of G/C nucleotides in SEQ ID NO:2;(iii) the nucleotide sequence does not contain the slice site GGTGAT (SEQ ID NO: 4) or GGTAAG (SEQ ID NO: 5);(iv) the nucleotide sequence contains fewer destabilizing element ATTTA (SEQ ID NO: 6) relative to SEQ ID NO:2;(v) the nucleotide sequence contains less repeat sequences relative to SEQ ID NO:2;(vi) the nucleotide sequence contains less antiviral motifs (SEQ ID NO: 11) relative to SEQ ID NO:2;(vii) the nucleotide sequence does not contain a poly-T sequence (SEQ ID NO: 7);(viii) the nucleotide sequence does not contain a poly-A sequence comprising AATAAA (SEQ ID NO: 9), ATTAAA (SEQ ID NO:10), AAAAAA (SEQ ID NO:8), or any combination thereof; or(xi) any combination of (i) to (viii).16. The isolated nucleic ...

CHARACTERIZATION OF PRE-REFINED CRUDE DISTILLATE FRACTIONS

Methods are provided for qualifying jet fuel fractions that are derived at least in part from pre-refined crude oil sources. The methods allow for determination of the stability of a jet fuel product over time by using an accelerated aging test. The methods are beneficial for verifying the stability of a jet fuel fraction that includes a portion derived from a pre-refined crude oil. 120.-. (canceled)21. A jet fuel product comprising:material derived from a pre-refined crude oil; wherein the material derived from a pre-refined crude oil is about 100% or less of a volume percentage of the jet fuel product;wherein the jet fuel product has a breakpoint of at least about 265° C. after an aging period equivalent to one year of aging at about 20° C.22. The jet fuel product of claim 21 , wherein the breakpoint of at least about 265° C. after an aging period equivalent to one year of aging at about 20° C. differs 10° C. or less from a breakpoint of the jet fuel product prior to the aging period.23. The jet fuel product of claim 21 , wherein the material derived from a pre-refined crude has an initial boiling point of at least about 284° F. (140° C.) and a final boiling point of less than about 572° F. (300° C.).24. The jet fuel product of claim 21 , wherein the material derived from a pre-refined crude oil is about 90% or less of a volume percentage of the jet fuel product.25. The jet fuel product of claim 21 , wherein the material derived from a pre-refined crude oil is about 75% or less of a volume percentage of the jet fuel product.26. The jet fuel product of claim 21 , wherein the material derived from a pre-refined crude oil is about 55% or less of a volume percentage of the jet fuel product.27. The jet fuel product of claim 21 , wherein the material derived from a pre-refined crude oil is about 50% or less of a volume percentage of the jet fuel product.28. The jet fuel product of claim 21 , wherein the material derived from a pre-refined crude oil is hydroprocessed.29. ...

Systems for Factor VIII Processing and Methods Thereof

The present invention provides methods of reducing nonprocessed Factor VIII or a chimeric polypeptide comprising Factor VIII comprising co-transfecting in a host cell a polynucleotide encoding Factor VIII with a polynucleotide encoding a protein convertase, where the endogenous processing enzymes of the host cell are insufficient to convert all of the Factor VIII to its processed isoform; expressing a proprotein convertase from a second polynucleotide in the host cell; and reducing the nonprocessed Factor VIII by processing with said proprotein convertase.

Immunoglobulin Chimeric Monomer-Dimer Hybrids

The invention relates to a chimeric monomer-dimer hybrid protein wherein said protein comprises a first and a second polypeptide chain, said first polypeptide chain comprising at least a portion of an immunoglobulin constant region and a biologically active molecule, and said second polypeptide chain comprising at least a portion of an immunoglobulin constant region without the biologically active molecule of the first chain. The invention also relates to methods of using and methods of making the chimeric monomer-dimer hybrid protein of the invention. 1. A chimeric protein comprising a first and second polypeptide chain , wherein said first chain comprises a biologically active molecule , and at least a portion of an immunoglobulin constant region and wherein said second chain comprises at least a portion of an immunoglobulin constant region without a biologically active molecule or immunoglobulin variable region.2. The chimeric protein of claim 1 , wherein said second chain further comprises an affinity tag.3. The chimeric protein of claim 2 , wherein the affinity tag is a FLAG tag.4. The chimeric protein of claim 1 , wherein the portion of an immunoglobulin is an Fc fragment.531.-. (canceled)32. A chimeric protein comprising a first and second polypeptide chaina) wherein said first chain comprises a biologically active molecule, at least a portion of an immunoglobulin constant region, and a first domain having at least one specific binding partner; andb) wherein said second chain comprises at least a portion of an immunoglobulin without a biologically active molecule or immunoglobulin variable region and further comprising a second domain said second domain being a specific binding partner of said first domain.33. The chimeric protein of claim 32 , wherein said second chain further comprises an affinity tag.34. The chimeric protein of claim 33 , wherein the affinity tag is a FLAG tag.35. The chimeric protein of claim 32 , wherein the portion of an immunoglobulin ...

Factor IX Polypeptides and Methods of Use Thereof

The present invention provides methods of administering Factor IX; methods of administering chimeric and hybrid polypeptides comprising Factor IX; polynucleotides encoding such chimeric and hybrid polypeptides; cells comprising such polynucleotides; and methods of producing such chimeric and hybrid polypeptides using such cells. 1. A method of administering Factor IX to a human subject in need thereof , comprising administering to the subject a dose of at least about 25 IU/kg of a chimeric polypeptide comprising Factor IX and a FcRn binding partner (FcRn BP) at about a once weekly or longer dosing interval.2. A method of administering Factor IX to a human subject in need thereof , comprising administering to the subject a dose of at least about 10 or at least about 20 IU/kg of a chimeric polypeptide comprising Factor IX and a FcRn binding partner (FcRn BP) at about a once weekly or longer dosing interval.3. A method of administering Factor IX to a human subject in need thereof , comprising administering to the subject a dose of at least about 10 IU/kg of a chimeric polypeptide comprising Factor IX and XTEN at about a once weekly or longer dosing interval.43. The method of any of - claims 1 , wherein the plasma level of said chimeric polypeptide reaches a trough of at least about 1 IU/dl after at least about 6 days in at least about 80% of a patient population or reaches a trough of at least about 1 IU/dl after at least about 6 days in said subject.53. The method of any of - claims 1 , wherein the plasma level of said chimeric polypeptide reachesan average trough of about 1-5 IU/dl in a patient population; ora trough of about 1-5 IU/dl in said subject.65. The method of any of - claims 1 , wherein less than 25% of the Factor IX chimeric polypeptide in said dose is fully phosphorylated and less than 25% of the Factor IX chimeric polypeptide in said dose is fully sulfated.7. The method of claim 6 , wherein less than about 10% of said chimeric polypeptide in said dose is ...

Factor IX Polypeptides and Methods of Use Thereof

The present invention provides methods of administering Factor IX; methods of administering chimeric and hybrid polypeptides comprising Factor IX; chimeric and hybrid polypeptides comprising Factor IX; polynucleotides encoding such chimeric and hybrid polypeptides; cells comprising such polynucleotides; and methods of producing such chimeric and hybrid polypeptides using such cells. 1121-. (canceled)122. A method of administering Factor IX (FIX) to a human subject in need thereof , comprising administering to the subject a dose of at least about 10 IU/kg of a long-acting FIX polypeptide at about a once weekly or longer dosing interval.123. The method of claim 122 , wherein the plasma level of said long-acting FIX polypeptide reaches a trough of at least about 1 IU/dl after at least about 6 days in said subject.124. The method of claim 122 , wherein said long-acting FIX polypeptide exhibits one or more characteristics selected from the group consisting of:a. an incremental recovery (K-Value) (activity; observed) greater than 0.75 IU/dL per IU/kg;b. an incremental recovery (K value) (activity; observed) of about 0.62-1.17 IU/dL per IU/kg;c. a clearance (CL) (activity) of about 1.84-4.58 mL/hour/kg;d. a clearance (CL) (activity) of about 3.36±0.93 mL/hour/kg;e. a mean residence time (MRT) (activity) of about 53.1-85.8 hours;f. a mean residence time (MRT) (activity) of about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, or about 90 hours;g. a MRT (activity) of at least about 68.05±11.16 hours;{'sub': '1/2beta', 'h. a t(activity) of about 40-67.4 hours;'}{'sub': '1/2beta', 'i. a t(activity) of at least about 40, about 45, about 50, about 55, about 60, about 65, about 70, or about 75 hours;'}{'sub': '1/2beta', 'j. a t(activity) of at least 75 hours;'}{'sub': 1/2beta', '1/2beta, 'k. a t t(activity) of about 3 fold longer than the t(activity) of a polypeptide consisting of FIX;'}l. a Vss (activity) of about 145-365 mL/kg;m. a Vss ( ...

Clotting Factor-Fc Chimeric Proteins to Treat Hemophilia

The invention relates to a chimeric protein comprising at least one clotting factor and at least a portion of an immunoglobulin constant region. The invention relates to a method of treating a hemostatic disorder comprising administering a therapeutically effective amount of a chimeric protein wherein the chimeric protein comprises at least one clotting factor and at least a portion of an immunoglobulin constant region. 142.-. (canceled)43. A nucleic acid molecule encoding a chimeric protein comprising a first polypeptide chain and a second polypeptide chain , the nucleic acid molecule comprising:(i) a first nucleic acid sequence encoding the first polypeptide chain, which comprises a clotting factor and at least a portion of an immunoglobulin constant region, which is a neonatal Fc Receptor (FcRn) binding partner, wherein the clotting factor is Factor VII or Factor VIIa; and(ii) a second nucleic acid sequence encoding the second polypeptide chain, which comprises at least a portion of an immunoglobulin constant region, which is a FcRn binding partner, wherein the second nucleic acid sequence does not encode the clotting factor.44. A vector comprising the nucleic acid molecule of .45. (canceled)46. (canceled)47. The nucleic acid molecule of claim 43 , wherein said portion of an immunoglobulin constant region in the first polypeptide chain or in the second polypeptide chain is a portion of an IgG.48. (canceled)49. A host cell comprising the vector of .50. The host cell of claim 49 , wherein said host cell is a mammalian cell.5158.-. (canceled)59. A host cell expressing a chimeric protein comprising a first polypeptide chain and a second polypeptide chain claim 49 , the host cell comprising:(i) a first vector comprising a nucleic acid sequence encoding the first polypeptide chain, which comprises a clotting factor and at least a portion of an immunoglobulin constant region, which is a neonatal Fc Receptor (FcRn) binding partner, wherein the clotting factor is Factor ...

Systems for Factor VIII Processing and Methods Thereof

The present invention provides methods of reducing nonprocessed Factor VIII or a chimeric polypeptide comprising Factor VIII comprising co-transfecting in a host cell a polynucleotide encoding Factor VIII with a polynucleotide encoding a protein convertase, where the endogenous processing enzymes of the host cell are insufficient to convert all of the Factor VIII to its processed isoform; expressing a proprotein convertase from a second polynucleotide in the host cell; and reducing the nonprocessed Factor VIII by processing with said proprotein convertase. 1. A method for decreasing nonprocessed Factor VIII in a culturing medium , comprising ,a) expressing Factor VIII from a first polynucleotide in a host cell, where the endogenous processing enzymes of the host cell are insufficient to convert all of the Factor VIII to its processed isoform;b) contacting said Factor VIII with an exogenous proprotein convertase; andc) decreasing the nonprocessed Factor VIII by processing with said proprotein convertase.2. The method of claim 1 , wherein said exogenous proprotein convertase is expressed from a second polypeptide in the host cell expressing said Factor VIII or is added to said culture medium.34-. (canceled)5. The method of claim 1 , wherein said Factor VIII is processed by cleaving Arginine corresponding to amino acid residue 1648 in SEQ ID NO: 6 (full-length Factor VIII) or amino acid residue 754 in SEQ ID NO: 2 (B domain-deleted Factor VIII).6. The method of claim 5 , wherein said processed Factor VIII comprises a Factor VIII heavy chain and a Factor VIII light chain claim 5 , wherein said heavy chain and light chain are associated by a non-covalent bond.7. The method of claim 1 , wherein the level of said nonprocessed Factor VIII is decreased such that said Factor VIII contains more than 75% claim 1 , 80% claim 1 , 85% claim 1 , 90% claim 1 , 95% claim 1 , and 100% of the processed Factor VIII after processing by said proprotein convertase.8. The method of claim 1 ...

CHARACTERIZATION OF PRE-REFINED CRUDE DISTILLATE FRACTIONS

Methods are provided for qualifying jet fuel fractions that are derived at least in part from pre-refined crude oil sources. The methods allow for determination of the stability of a jet fuel product over time by using an accelerated aging test. The methods are beneficial for verifying the stability of a jet fuel fraction that includes a portion derived from a pre-refined crude oil. 1. A method for preparing a jet fuel or kerosene product , comprising:determining a breakpoint for a first sample of a distillate fraction, the distillate fraction having an initial boiling point of at least about 284° F. (140° C.) and a final boiling point of about 572° F. (300° C.) or less, at least a portion of the distillate fraction being derived from a first pre-refined crude oil;maintaining a second sample of the distillate fraction at a temperature of at least about 40° C. for an aging period;determining a breakpoint for the aged second sample of the distillate fraction, the breakpoint for the aged second sample being at least about 265° C.; andpreparing a jet fuel product comprising a kerosene portion derived from a second pre-refined crude oil, the second pre-refined crude oil being derived from the same source as the first pre-refined crude oil, a volume percentage of the kerosene portion derived from the second pre-refined crude in the jet fuel product being about 110% or less of a volume percentage corresponding to the portion of the distillate fraction derived from the first pre-refined crude oil, the initial boiling point of the jet fuel product being at least about the initial boiling point of the distillate fraction, and the final boiling point of the jet fuel product being less than or equal to the final boiling point of the distillate fraction.2. The method of claim 1 , further comprising:obtaining a portion of the distillate fraction; andsplitting the portion of the distillate fraction to form at least the first sample and the second sample.3. The method of claim 1 , ...

Methods of Using FIX Polypeptides

The present invention provides methods of administering long-acting Factor IX; methods of administering long-acting, chimeric and hybrid polypeptides comprising Factor IX; and methods of producing such chimeric and hybrid polypeptides using cells.

Immunoglobulin chimeric monomer-dimer hybrids

The invention relates to a chimeric monomer-dimer hybrid protein wherein the protein comprises a first and a second polypeptide chain, the first polypeptide chain comprising at least a portion of an immunoglobulin constant region and a biologically active molecule, and the second polypeptide chain comprising at least a portion of an immunoglobulin constant region without the biologically active molecule of the first chain. The invention also relates to methods of using and methods of making the chimeric monomer-dimer hybrid protein of the invention.

FACTOR IX POLYPEPTIDES AND METHODS OF USE THEREOF

The present invention provides methods of administering Factor IX; methods of administering chimeric and hybrid polypeptides comprising Factor IX; polynucleotides encoding such chimeric and hybrid polypeptides; cells comprising such polynucleotides; and methods of producing such chimeric and hybrid polypeptides using such cells. 1121-. (canceled)122. A method of on-demand treatment of bleeding , comprising intravenously administering to a hemophilia B human subject in need thereof a dose of about 25 IU/kg to about 100 IU/kg of a chimeric factor IX (FIX) polypeptide comprising human FIX having an amino acid sequence identical to amino acids 1 to 415 of SEQ ID NO: 2 and a FcRn binding partner (FcRn BP) , wherein the FcRn BP is human Fc or human albumin.123. The method of claim 122 , wherein the human subject exhibits a plasma FIX level of about 30 IU/dL to about 60 IU/dL following a dose of the chimeric FIX polypeptide.124. The method of claim 122 , wherein the human subject exhibits a plasma FIX level of about 80 IU/dL to about 100 IU/dL following a dose of the chimeric FIX polypeptide.125. The method of claim 122 , wherein the human subject exhibits a plasma FIX level about 20% to about 50% of normal following a dose of the chimeric FIX polypeptide.126. The method of claim 122 , wherein the human subject exhibits a plasma FIX level about 50% to about 100% of normal following a dose of the chimeric FIX polypeptide.127. The method of claim 123 , wherein the plasma FIX level is measured 8-12 hours following a dose of the chimeric FIX polypeptide.128. The method of claim 124 , wherein the plasma FIX level is measured 8-12 hours following a dose of the chimeric FIX polypeptide.129. The method of claim 125 , wherein the plasma FIX level is measured 8-12 hours following a dose of the chimeric FIX polypeptide.130. The method of claim 126 , wherein the plasma FIX level is measured 8-12 hours following a dose of the chimeric FIX polypeptide.131. The method of claim 122 , ...

CHIMERIC FACTOR VIII POLYPEPTIDES AND USES THEREOF

The present invention provides a VWF fragment comprising the D′ domain and D3 domain of VWF, a chimeric protein comprising the VWF fragment and a heterologous moiety, or a chimeric protein comprising the VWF fragment and a FVIII protein and methods of using the same. A polypeptide chain comprising a VWF fragment of the invention binds to or is associated with a polypeptide chain comprising a FVIII protein and the polypeptide chain comprising the VWF fragment can prevent or inhibit binding of endogenous VWF to the FVIII protein. By preventing or inhibiting binding of endogenous VWF to the FVIII, which is a half-life limiting factor for FVIII, the VWF fragment can induce extension of half-life of the FVIII protein. The invention also includes nucleotides, vectors, host cells, methods of using the VWF fragment, or the chimeric proteins. 1. A chimeric protein comprising a Factor VIII (“FVIII”) protein and an adjunct moiety (AM) , which are linked by a covalent bond , wherein the adjunct moiety inhibits or prevents endogenous VWF from binding to the FVIII protein.2. The chimeric protein of claim 1 , wherein the covalent bond prevents dissociation of the adjunct moiety from the FVIII protein in the presence of endogenous VWF.3. The chimeric protein of or claim 1 , wherein the covalent bond is a peptide bond.4. The chimeric protein of any one of to claim 1 , wherein the covalent bond is a disulfide bond.5. The chimeric protein of any one of to claim 1 , wherein the covalent bond is a linker between the FVIII protein and the adjunct moiety.65. The chimeric protein of any one of - claims 1 , wherein the adjunct moiety prevents the FVIII protein from being cleared through a VWF clearance pathway.7. The chimeric protein of claim any one of to claims 1 , wherein the adjunct moiety inhibits or prevents endogenous VWF from binding to the FVIII protein by shielding or blocking a VWF binding site on the FVIII protein.8. The chimeric protein of claim 7 , wherein the VWF binding site ...

FACTOR IX POLYPEPTIDES AND METHODS OF USE THEREOF

The present invention provides methods of administering Factor IX; methods of administering chimeric and hybrid polypeptides comprising Factor IX; polynucleotides encoding such chimeric and hybrid polypeptides; cells comprising such polynucleotides; and methods of producing such chimeric and hybrid polypeptides using such cells. 1. A method of reducing the frequency of spontaneous bleeding comprising intravenously administering to a hemophilia B human subject in need thereof multiple doses of about 50 IU/kg to about 100 IU/kg of a chimeric factor IX (“FIX”) polypeptide comprising human FIX having an amino acid sequence identical to amino acids 1 to 415 of SEQ ID NO:2 and an FcRn binding partner (“FcRn BP”) at a dosing interval of about 10 days to about 14 days between two doses , wherein the FcRn BP is human Fc or human albumin.2. The method of claim 1 , wherein the dosing interval is 14 days and each of the multiple doses is 50 IU/kg.3. The method of claim 1 , wherein the dosing interval is 14 days and each of the multiple doses is 60 IU/kg.4. The method of claim 1 , wherein the dosing interval is 14 days and each of the multiple doses is 70 IU/kg.5. The method of claim 1 , wherein the dosing interval is 14 days and each of the multiple doses is 80 IU/kg.6. The method of claim 1 , wherein the dosing interval is 14 days and each of the multiple doses is 90 IU/kg.7. The method of claim 1 , wherein the dosing interval is 14 days and each of the multiple doses is 100 IU/kg.8. The method of claim 1 , wherein the FcRn BP is human Fc.9. The method of claim 8 , wherein the human Fc comprises amino acids 1 to 227 of SEQ ID NO: 4.10. The method of claim 9 , wherein the subject exhibits the plasma FIX activity above 1 IU/dL during the dosing interval as measured by a one stage clotting assay that determines activated partial thromboplastin time.11. The method of claim 1 , wherein the chimeric FIX polypeptide further comprises a linker joining the FIX and the FcRN BP.12. A ...

OPTIMIZED FACTOR VIII GENE

The present invention provides codon optimized Factor VIII sequences, vectors and host cells comprising codon optimized Factor VIII sequences, polypeptides encoded by codon optimized Factor VIII sequences, and methods of producing such polypeptides. 1. An isolated nucleic acid molecule comprising a nucleotide sequence having at least 85% sequence identity to SEQ ID NO:1 , wherein the nucleotide sequence encodes a polypeptide with Factor VIII activity.2. The isolated nucleic acid molecule of claim 1 , wherein the nucleotide sequence has at least 90% claim 1 , at least 95% claim 1 , at least 96% claim 1 , at least 97% claim 1 , at least 98% claim 1 , at least 99% claim 1 , or 100% sequence identity to SEQ ID NO:1 claim 1 , optionally wherein:the nucleotide sequence comprises SEQ ID NO: 1;the human codon adaptation index is increased relative to SEQ ID NO:3;the human codon adaptation index is at least about 0.75, at least about 0.76, at least about 0.77, at least about 0.78, at least about 0.79, at least about 0.80, at least about 0.81, at least about 0.82, at least about 0.83, at least about 0.84, at least about 0.85, at least about 0.86, at least about 0.87, or at least about 0.88;the nucleotide sequence contains a higher percentage of G/C nucleotides compared to the percentage of G/C nucleotides in SEQ ID NO:3, optionally wherein the percentage of G/C nucleotides is at least about 45%, at least about 46%, at least about 47%, at least about 48%, at least about 49%, or at least about 50%;the nucleotide sequence contains fewer MARS/ARS sequences (SEQ ID NO:5 and SEQ ID NO:6) relative to SEQ ID NO:3, optionally wherein the nucleotide sequence contains at most one MARS/ARS sequence; or the nucleotide sequence does not contain a MARS/ARS sequence;the nucleotide sequence does not contain the splice site GGTGAT (SEQ ID NO:7);the nucleotide sequence contains fewer destabilizing elements (SEQ ID NO:8 and SEQ ID NO:9) relative to SEQ ID NO:3, optionally wherein:the nucleotide ...

FACTOR IX FUSION PROTEINS AND METHODS OF MAKING AND USING SAME

The present invention provides Factor IX (FIX) fusion proteins comprising at least one heterologous moiety, such as an XTEN. The present invention further discloses methods of making and using the FIX fusion proteins. 1. A Factor IX (FIX) fusion protein comprising a FIX polypeptide and an XTEN that is inserted within the FIX polypeptide , wherein the FIX fusion protein comprises the FIX polypeptide amino acid sequence of SEQ ID NO: 2 with the XTEN inserted at an insertion site in the activation peptide of SEQ ID NO: 2 (amino acids 146-180) , and wherein the FIX fusion protein exhibits procoagulant activity.2. The FIX fusion protein of claim 1 , wherein the insertion site corresponds to an amino acid selected from the group consisting of amino acid 149 of SEQ ID NO: 2 claim 1 , amino acid 162 of SEQ ID NO: 2 claim 1 , amino acid 166 of SEQ ID NO: 2 claim 1 , amino acid 174 of SEQ ID NO: 2 and any combination thereof.3. The FIX fusion protein of claim 1 , wherein the XTEN comprises at least about 5 amino acids claim 1 , at least about 6 amino acids claim 1 , at least about 12 amino acids claim 1 , at least about 36 amino acids claim 1 , at least about 42 amino acids claim 1 , at least about 72 amino acids claim 1 , at least about 144 amino acids claim 1 , or at least about 288 amino acids.4. The FIX fusion protein of claim 1 , which further comprises a second XTEN.5. The FIX fusion protein of claim 4 , wherein the XTEN is inserted within the FIX polypeptide at an insertion site corresponding to amino acid 166 of SEQ ID NO: 2 claim 4 , and wherein the second XTEN is fused to the C-terminus of the FIX polypeptide.6. The FIX fusion protein of claim 1 , further comprising a first Fc domain.7. The FIX fusion protein of claim 6 , further comprising a second Fc domain.8. The FIX fusion protein of claim 7 , which comprises two polypeptide chains claim 7 , wherein(i) the first polypeptide chain comprises the first Fc domain fused to the N-terminus or C-terminus of the FIX ...

FACTOR VIII COMPLEX WITH XTEN AND VON WILLEBRAND FACTOR PROTEIN, AND USES THEREOF

The present invention provides a chimeric protein comprising a VWF protein comprising the D′ domain and D3 domain of VWF, one or more XTEN sequence, and a FVIII protein, wherein the VWF fragment, the XTEN sequence, or the FVIII protein are linked to or associated with each other. The chimeric protein can further comprise one or more Ig constant region or a portion thereof (e.g., an Fc region). A polypeptide chain comprising a VWF fragment of the invention binds to or is associated with a polypeptide chain comprising a FVIII protein linked to an XTEN sequence and the polypeptide chain comprising the VWF fragment can prevent or inhibit binding of endogenous VWF to the FVIII protein linked to the XTEN sequence. By preventing or inhibiting binding of endogenous VWF to the FVIII protein, which is a half-life limiting factor for FVIII, the VWF fragment can induce extension of half-life of the chimeric protein comprising a FVIII protein. The invention also includes nucleotides, vectors, host cells, methods of using the VWF fragment, or the chimeric proteins. 1. A chimeric protein comprising (i) a von Willebrand Factor (VWF) protein comprising a D′ domain and a D3 domain of VWF , (ii) an XTEN sequence , and (iii) a FVIII protein , wherein the VWF fragment and the XTEN sequence are linked by an optional linker , wherein the VWF fragment or the XTEN sequence is linked to or associated with the FVIII protein.3. The chimeric protein of claim 1 , wherein the XTEN sequence is linked to the FVIII protein by a linker.412-. (canceled)13. The chimeric protein of claim 1 , further comprising a first Ig constant region or a portion thereof.1416-. (canceled)17. The chimeric protein of claim 13 , wherein the chimeric protein comprises a second Ig constant region claim 13 , wherein the second Ig constant region is linked to the VWF fragment by a linker.1819-. (canceled)20. The chimeric protein of claim 17 , wherein the second Ig constant region or a portion thereof is associated with the ...

Clotting Factor-Fc Chimeric Proteins to Treat Hemophilia

The Invention relates to a chimeric protein comprising at least one clotting factor and at least a portion of an immunoglobulin constant region. The invention relates to a method of treating a hemostatic disorder comprising administering a therapeutically effective amount of a chimeric protein wherein the chimeric protein comprises at least one clotting factor and at least a portion of an immunoglobulin constant region. 158-. (canceled)59. A chimeric protein comprising a first polypeptide and a second polypeptide , wherein the first polypeptide comprises (i) a clotting factor , which is Factor VIII , Factor VIIIa , Factor IX , Factor IXa , Factor VII , or Factor VIIa , and (ii) at least a portion of an immunoglobulin constant region fused to the clotting factor , which is a neonatal Fc Receptor (FcRn) binding partner , andthe second polypeptide comprises at least a portion of an immunoglobulin constant region, which is a FcRn binding partner, without the clotting factor of the first polypeptide and without an immunoglobulin variable domain, andwherein the first polypeptide and the second polypeptide are linked.60. The chimeric protein of claim 59 , wherein the clotting factor is fused to the portion of an immunoglobulin constant region by a linker.61. The chimeric protein of claim 60 , wherein the linker comprises about 1 to about 20 amino acids.62. The chimeric protein of claim 60 , wherein the linker comprises the sequence (GlyGlySer)n or SEQ ID NO: 31 claim 60 , wherein n is an integer selected from the group consisting of 1 claim 60 , 2 claim 60 , 3 claim 60 , 4 claim 60 , 5 claim 60 , 6 claim 60 , 7 claim 60 , 8 claim 60 , 9 claim 60 , and 10.63. The chimeric protein of claim 59 , wherein the portion of an immunoglobulin constant region of the first polypeptide is an Fc fragment.64. The chimeric protein of claim 63 , wherein the portion of an immunoglobulin constant region of the second polypeptide is an Fc fragment.65. The chimeric protein of claim 63 , ...

METHODS OF USING FIX POLYPEPTIDES

The present invention provides methods of administering long-acting Factor IX; methods of administering long-acting, chimeric and hybrid polypeptides comprising Factor IX; and methods of producing such chimeric and hybrid polypeptides using cells.

NUCLEIC ACID MOLECULES AND USES THEREOF FOR NON-VIRAL GENE THERAPY

The present disclosure provides nucleic acid molecules comprising a first inverted terminal repeat (ITR), a second ITR, and a genetic cassette encoding a target sequence. In some embodiments, the target sequence encodes a miRNA and/or a therapeutic protein. In certain embodiments, the therapeutic protein comprises a clotting factor, a growth factor, a hormone, a cytokine, an antibody, a fragment thereof, and a combination thereof. In some embodiments, the first ITR and/or the second ITR is an ITR of a non-adeno-associated virus (AAV). The present disclosure also provides methods of treating a metabolic disorder of the liver in a subject comprising administering to the subject the nucleic acid molecule or a polypeptide encoded thereby. 1. A nucleic acid molecule comprising a first inverted terminal repeat (ITR) and a second ITR flanking a genetic cassette comprising a heterologous polynucleotide sequence , wherein the first ITR and/or second ITR comprises a nucleotide sequence at least about 75% , at least about 80% , at least about 85% , at least about 90% , at least about 95% , at least about 96% , at least about 97% , at least about 98% , at least about 99% , or 100% identical to a nucleotide sequence set forth in SEQ ID NO: 180 , 181 , 183 , 184 , 185 , 186 , 187 or 188 , or a functional derivative thereof.2. The nucleic acid molecule of claim 1 , optionally wherein:the first ITR comprises the nucleotide sequence set forth in SEQ ID NO: 180 and the second ITR comprises the nucleotide sequence set forth in SEQ ID NO: 181;the first ITR comprises the nucleotide sequence set forth in SEQ ID NO: 183 and the second ITR comprises the nucleotide sequence set forth in SEQ ID NO: 184;the first ITR comprises the nucleotide sequence set forth in SEQ ID NO: 185 and the second ITR comprises the nucleotide sequence set forth in SEQ ID NO: 186;the first ITR comprises the nucleotide sequence set forth in SEQ ID NO: 187 and the second ITR comprises the nucleotide sequence set ...

Immunoglobulin chimeric monomer-dimer hybrids

The invention relates to a chimeric monomer-dimer hybrid protein wherein the protein comprises a first and a second polypeptide chain, the first polypeptide chain comprising at least a portion of an immunoglobulin constant region and a biologically active molecule, and the second polypeptide chain comprising at least a portion of an immunoglobulin constant region without the biologically active molecule of the first chain. The invention also relates to methods of using and methods of making the chimeric monomer-dimer hybrid protein of the invention.

CHIMERIC CLOTTING FACTORS

The invention provides chimeric clotting factors comprising an activatable clotting factor and an enhancer moiety. The activatable clotting factor allows the chimeric clotting factor to be activated at the site of coagulation. The enhancer moiety can additionally improve procoagulation activities of the chimeric clotting factors. The chimeric clotting factors can further be improved by fusion to a half-life extender, which improves a pharmacokinetics property of the chimeric clotting factor. The invention also includes methods of making and methods of using these chimeric clotting factors.

FACTOR VIII CHIMERIC PROTEINS AND USES THEREOF

The present invention provides a chimeric protein comprising a first polypeptide which comprises a FVIII protein and a first Ig constant region or a portion thereof and a second polypeptide which comprises a VWF protein comprising the D′ domain and D3 domain of VWF, a XTEN sequence having less than 288 amino acids in length, and a second Ig constant region or a portion thereof, wherein the first polypeptide and the second polypeptide are associated with each other. The invention also includes nucleotides, vectors, host cells, methods of using the chimeric proteins. 1149-. (canceled)151. The method of claim 150 , wherein wherein the first first Ig constant region or portion thereof is associated with the second Ig constant region or portion thereof through two disulfide bonds.152. The method of claim 151 , wherein wherein the first first Ig constant region or portion thereof is a first Fc region and the second Ig constant region or portion thereof is a second Fc region.153. The method of claim 152 , wherein the cleavable linker is 20 to 50 amino acids long.154. The method of claim 151 , wherein the cleavable linker is about 30 amino acids long.155. The method of claim 150 , wherein the hemostatic disorder is hemophilia A.156. The method of claim 150 , wherein the chimeric protein is administered prophylactically.157. The method of claim 150 , wherein the chimeric protein is administered for on-demand treatment.158. The method of claim 150 , wherein the chimeric protein is administered prior to claim 150 , during claim 150 , or after surgery.159. The method of claim 150 , wherein the chimeric protein is administered to control an acute bleeding episode.160. The method of claim 150 , wherein the chimeric protein is administered intravenously or subcutaneously.161. The method of claim 160 , wherein the chimeric protein is administered intravenously.162. A chimeric protein comprising: (a) a Factor VIII (“FVIII”) protein comprising a N-terminal portion, fused to a first ...

Immunoglobulin Chimeric Monomer-Dimer Hybrids

The invention relates to a chimeric monomer-dimer hybrid protein wherein the protein comprises a first and a second polypeptide chain, the first polypeptide chain comprising at least a portion of an immunoglobulin constant region and a biologically active molecule, and the second polypeptide chain comprising at least a portion of an immunoglobulin constant region without the biologically active molecule of the first chain. The invention also relates to methods of using and methods of making the chimeric monomer-dimer hybrid protein of the invention. 1194.-. (canceled)195. A pharmaceutical composition comprising (a) a chimeric protein which comprises a first polypeptide and a second polypeptide and (b) a pharmaceutically acceptable carrier , wherein the first polypeptide comprises (i) a biologically active molecule selected from a hormone , a cytokine , a growth factor , and a cell surface receptor and (ii) an immunoglobulin constant region or a portion thereof comprising a neonatal Fc Receptor binding site , and the second polypeptide comprises an immunoglobulin constant region or a portion thereof comprising a neonatal Fc Receptor binding site and does not comprise the biologically active molecule of (i) or an immunoglobulin variable domain , and wherein the first polypeptide and the second polypeptide are linked.196. The pharmaceutical composition of claim 195 , wherein the immunoglobulin constant region or a portion thereof in the first polypeptide is an Fc fragment.197. The pharmaceutical composition of claim 196 , wherein the immunoglobulin constant region or a portion thereof in the second polypeptide is an Fc fragment.198. The pharmaceutical composition of claim 195 , wherein the immunoglobulin constant region or a portion thereof in the first polypeptide and the immunoglobulin constant region or a portion thereof in the second polypeptide are identical.199. The pharmaceutical composition of claim 195 , wherein the first polypeptide comprises the hormone and ...

NUCLEIC ACID MOLECULES AND USES THEREOF

The present disclosure provides nucleic acid molecules comprising a first inverted terminal repeat (ITR), a second ITR, and a genetic cassette encoding a miRNA and/or a therapeutic protein. In certain embodiments, the therapeutic protein comprises a clotting factor, e.g., a FVIII polypeptide, a FIX polypeptide, or a fragment thereof. In some embodiments, the first ITR and/or the second ITR is an ITR of a non-adeno-associated virus (AAV). The present disclosure also provides methods of treating bleeding disorders such as hemophilia comprising administering to the subject the nucleic acid molecule or a polypeptide encoded thereby. 1. A nucleic acid molecule comprising a first inverted terminal repeat (ITR) , a second ITR , and a genetic cassette;wherein the first ITR and/or the second ITR are an ITR of a non-adeno-associated virus (non-AAV), wherein the genetic cassette is positioned between the first ITR and the second ITR, and wherein the genetic cassette encodes a therapeutic protein, a miRNA, or both a therapeutic protein and a miRNA.2. The nucleic acid molecule of claim 1 , wherein the therapeutic protein comprises a clotting factor.3. The nucleic acid molecule of or claim 1 , wherein the non-AAV is selected from the group consisting of a member of the viral family Parvoviridae.4. The nucleic acid molecule of any one of to claim 1 , wherein the first ITR and the second ITR are ITRs of a non-AAV.5. The nucleic acid molecule of or claim 1 , wherein the member of the viral family Parvoviridae is selected from the group consisting of Bocavirus claim 1 , Dependovirus claim 1 , Erythrovirus claim 1 , Amdovirus claim 1 , Parvovirus claim 1 , Densovirus claim 1 , Iteravirus claim 1 , Contravirus claim 1 , Aveparvovirus claim 1 , Copiparvovirus claim 1 , Protoparvovirus claim 1 , Tetraparvovirus claim 1 , Ambidensovirus claim 1 , Brevidensovirus claim 1 , Hepandensovirus claim 1 , Penstyldensovirus Muscovy duck parvovirus (MDPV) strain claim 1 , porcine parvovirus (U44978 ...

Factor IX Polypeptides and Methods of Use Thereof

The present invention provides methods of administering Factor IX; methods of administering chimeric and hybrid polypeptides comprising Factor IX; chimeric and hybrid polypeptides comprising Factor IX; polynucleotides encoding such chimeric and hybrid polypeptides; cells comprising such polynucleotides; and methods of producing such chimeric and hybrid polypeptides using such cells. 1121-. (canceled)122. A method of increasing the level of Factor IX activity in a subject's plasma comprising administering to the subject multiple doses of about 25 IU/kg to about 125 IU/kg of a long-acting FIX polypeptide comprising FIX and an FcRn binding partner (“FcRn BP”) at a dosing interval of about 7 days to about 24 days between two doses.123. The method of claim 122 , wherein each of the multiple doses is 30 IU/kg to 40 IU/kg claim 122 , 40 IU/kg to 50 IU/kg claim 122 , 50 IU/kg to 60 IU/kg claim 122 , 60 IU/kg to 70 IU/kg claim 122 , 70 IU/kg to 80 IU/kg claim 122 , 80 IU/kg to 90 IU/kg claim 122 , 90 IU/kg to 100 IU/kg claim 122 , 100 IU/kg to 110 IU/kg. or 110 IU/kg to 120 IU/kg.124. The method of claim 122 , wherein each of the multiple doses is 40 IU/kg to 50 IU/kg.125. The method of claim 122 , wherein each of the multiple doses is 70 IU/kg to 80 IU/kg.126. The method of claim 122 , wherein each of the multiple doses is 90 IU/kg to 100 IU/kg.127. The method of claim 122 , wherein the dosing interval is 7 days to 14 days.128. The method of claim 122 , wherein the dosing interval is 9 days to 18 days.129. The method of claim 122 , wherein the dosing interval is 19 days claim 122 , 20 days claim 122 , 21 days claim 122 , 22 days claim 122 , 23 days claim 122 , or 24 days.130. The method of claim 124 , wherein the dosing interval is 7 days to 14 days.131. The method of claim 125 , wherein the dosing interval is 9 days to 18 days.132. The method of claim 126 , wherein the dosing interval is 19 days to 20 days. 21 days claim 126 , 22 days claim 126 , 23 days claim 126 , or 24 ...

FACTOR VIII COMPLEX WITH XTEN AND VON WILLEBRAND FACTOR PROTEIN, AND USES THEREOF

The present invention provides a chimeric protein comprising a VWF protein comprising the D′ domain and D3 domain of VWF, one or more XTEN sequence, and a FVIII protein, wherein the VWF fragment, the XTEN sequence, or the FVIII protein are linked to or associated with each other. The chimeric protein can further comprise one or more Ig constant region or a portion thereof (e.g., an Fc region). A polypeptide chain comprising a VWF fragment of the invention binds to or is associated with a polypeptide chain comprising a FVIII protein linked to an XTEN sequence and the polypeptide chain comprising the VWF fragment can prevent or inhibit binding of endogenous VWF to the FVIII protein linked to the XTEN sequence. By preventing or inhibiting binding of endogenous VWF to the FVIII protein, which is a half-life limiting factor for FVIII, the VWF fragment can induce extension of half-life of the chimeric protein comprising a FVIII protein. The invention also includes nucleotides, vectors, host cells, methods of using the VWF fragment, or the chimeric proteins. 1119-. (canceled)120. A polynucleotide or a set of polynucleotides encoding a chimeric protein comprising (i) a von Willebrand Factor (VWF) protein comprising the D′ domain and the D3 domain of VWF , (ii) an XTEN sequence , and (iii) a FVIII protein , wherein the VWF fragment and the XTEN sequence are linked by an optional linker , wherein the VWF fragment or the XTEN sequence is linked to or associated with the FVIII protein.121. The polynucleotide of claim 120 , further comprising a polynucleotide chain claim 120 , which encodes PC5 or PC7.122. A vector comprising the polynucleotide of and one or more promoter operably linked to the polynucleotide or the set of polynucleotides.123. The vector of claim 122 , further comprising an additional vector claim 122 , which comprises a polynucleotide chain encoding PC5 or PC7.124. A host cell comprising the polynucleotide of any one of .125. The host cell of claim 124 , which ...

FACTOR VIII CHIMERIC AND HYBRID POLYPEPTIDES, AND METHODS OF USE THEREOF

The present invention provides methods of administering Factor VIII (processed FVIII, single chain FVIII, or a combination thereof); methods of administering chimeric and hybrid polypeptides comprising Factor VIII; chimeric and hybrid polypeptides comprising Factor VIII; polynucleotides encoding such chimeric and hybrid polypeptides; cells comprising such polynucleotides; and methods of producing such chimeric and hybrid polypeptides using such cells. 1. A method of administering Factor VIII to a human subject in need thereof , comprising administering to the subject a therapeutic dose of a chimeric polypeptide comprising a Factor VIII portion and a second portion , wherein said Factor VIII portion comprises processed Factor VIII , which has two chains , a first chain comprising a heavy chain and a second chain comprising a light chain , wherein about 50% , about 60% , about 70% , about 75% , about 80% , about 85% , about 90% , about 95% , or about 99% of the Factor VIII portion is processed Factor VIII , and wherein said first chain and said second chain are associated by a metal bond.2. (canceled)3. (canceled)4. The method of claim 1 , wherein said Factor VIII portion comprises single chain Factor VIII claim 1 , which is a single chain.5. (canceled)6. (canceled)7. The method of claim 1 , wherein at least about 1% claim 1 , about 5% claim 1 , about 10% claim 1 , about 15% claim 1 , about 20% claim 1 , about 25% claim 1 , about 30% claim 1 , about 35% claim 1 , or about 40% of the Factor VIII portion of the chimeric polypeptide is single chain Factor VIII.835-. (canceled)36. The method of claim 1 , wherein said chimeric polypeptide exhibits one or more pharmacokinetic parameters claim 1 , in said subject claim 1 , selected from the group consisting of:{'sub': max_', 'max_, 'a COBS in said subject administered with 25 IU/kg of the chimeric polypeptide is comparable to the COBS in a subject administered with 25 IU/kg of a polypeptide consisting of the full-length, ...

FACTOR VIII-XTEN FUSIONS AND USES THEREOF

The present invention provides chimeric proteins comprising a Factor VIII (FVIII) polypeptide and XTEN polypeptides. The FVIII polypeptide can have a reduced affinity for von Willebrand Factor (VWF). XTEN polypeptides of appropriate sizes are inserted into the FVIII polypeptide at particular locations in order to extend the half-life of the FVIII polypeptide. The invention also includes nucleotides, vectors, host cells, and methods of using the chimeric proteins, e.g., to treat bleeding diseases and disorders. 1. A chimeric protein comprising (i) a Factor VIII (FVIII) polypeptide and (ii) an XTEN sequence wherein the FVIII polypeptide has reduced affinity for von Willebrand Factor (VWF).2. The chimeric protein of comprising at least four XTEN sequences.3. A chimeric protein comprising (i) a FVIII polypeptide and (ii) at least four XTEN sequences.4. The chimeric protein of or wherein an XTEN sequences is inserted in the A1 domain of the FVIII polypeptide claim 1 , between the A2 and A3 domains of the FVIII polypeptide claim 1 , in the A3 domain of the FVIII polypeptide claim 1 , and at the C terminus of the VIII polypeptide.5. The chimeric protein of any one of - claim 1 , wherein at least one of the at least four XTEN sequences is no more than 72 amino acids in length.6. The chimeric protein of claim 5 , wherein at least one of the at least four XTEN sequences is 36 or 42 amino acids in length.7. The chimeric protein of any one of - comprising a spacer between an amino acid in the FVIII polypeptide an amino acid in the XTEN.8. The chimeric protein of claim 7 , wherein the spacer comprises a cleavage sequence or amino acids encoded by polynucleotides with a restriction site.9. The chimeric protein of any one of - claim 7 , wherein the FVIII polypeptide lacks amino acids 745-1685 of full-length mature FVIII polypeptide (SEQ ID NO: 4).10. The chimeric protein of any one of - claim 7 , wherein the FVIII polypeptide lacks amino acids 741-1689 of full-length mature FVIII ...

FACTOR IX POLYPEPTIDES AND METHODS OF USE THEREOF

The present invention provides methods of administering Factor IX; methods of administering chimeric and hybrid polypeptides comprising Factor IX; polynucleotides encoding such chimeric and hybrid polypeptides; cells comprising such polynucleotides; and methods of producing such chimeric and hybrid polypeptides using such cells. 1. A method of reducing the frequency of spontaneous bleeding , comprising intravenously administering to a human hemophilia B subject in need thereof multiple doses of about 25 IU/kg to about 50 IU/kg of a chimeric Factor IX (“FIX”) polypeptide comprising human FIX having an amino acid sequence identical to amino acids 1 to 415 of SEQ ID NO:2 and an FcRn binding partner (“FcRn BP”) at a dosing interval of about 7 days between two doses , wherein the FcRn BP is human Fc or human albumin.2. The method of claim 1 , wherein the FcRn BP is human albumin.3. The method of claim 2 , wherein the human albumin has an amino acid sequence identical to SEQ ID NO: 5.4. The method of claim 3 , wherein the chimeric FIX polypeptide further comprises a linker joining the FIX and the human albumin.5. The method of claim 4 , wherein the linker has an amino acid sequence SEQ ID NO: 6.6. The method of claim 5 , wherein the linker joins the FIX to the N-terminal end of the human albumin.7. The method of claim 6 , wherein the dosing interval is 7 days.8. The method of claim 7 , wherein each of the multiple doses is 25 IU/kg to 40 IU/kg.9. The method of claim 1 , wherein the subject exhibits the plasma FIX activity above 1 IU/dL during the dosing interval as measured by a one stage clotting assay that determines activated partial thromboplastin time.10. The method of claim 9 , wherein the FcRn BP is human albumin.11. The method of claim 10 , wherein the human albumin has an amino acid sequence identical to SEQ ID NO: 5.12. The method of claim 11 , wherein the chimeric FIX polypeptide further comprises a linker joining the FIX and the human albumin.13. The method of ...

FACTOR IX POLYPEPTIDES AND METHODS OF USE THEREOF

The present invention provides methods of administering Factor IX; methods of administering chimeric and hybrid polypeptides comprising Factor IX; polynucleotides encoding such chimeric and hybrid polypeptides; cells comprising such polynucleotides; and methods of producing such chimeric and hybrid polypeptides using such cells. 1. A method of treating hemophilia B comprising intravenously administering to a human subject in need thereof multiple doses of about 50 IU/kg to about 100 IU/kg of a chimeric factor IX (“FIX”) polypeptide comprising human FIX having an amino acid sequence identical to amino acids 1 to 415 of SEQ ID NO:2 and an FcRn binding partner (“FcRn BP”) at a dosing interval of about 10 days to about 14 days between two doses , wherein the FcRn BP is human Fc or human albumin , wherein the administration treats the human subject by reducing the frequency of spontaneous bleeding.2. The method of claim 1 , wherein the FcRn BP is human albumin.3. The method of claim 2 , wherein the human albumin has an amino acid sequence identical to SEQ ID NO: 5.4. The method of claim 3 , wherein the chimeric FIX polypeptide further comprises a linker joining the FIX and the human albumin.5. The method of claim 4 , wherein the linker has an amino acid sequence SEQ ID NO: 6.6. The method of claim 5 , wherein the linker joins the FIX to the N-terminal end of the human albumin.7. The method of claim 6 , wherein the dosing interval is 14 days.8. The method of claim 7 , wherein the subject exhibits the plasma FIX activity above 1 IU/dL during the dosing interval as measured by a one stage clotting assay that determines activated partial thromboplastin time.9. The method of claim 8 , wherein the FcRn BP is human albumin.10. The method of claim 9 , wherein the human albumin has an amino acid sequence identical to SEQ ID NO: 5.11. The method of claim 10 , wherein the chimeric FIX polypeptide further comprises a linker joining the FIX and the human albumin.12. The method of ...

Factor IX Polypeptides and Methods of Use Thereof

The present invention provides methods of administering Factor IX; methods of administering chimeric and hybrid polypeptides comprising Factor IX; polynucleotides encoding such chimeric and hybrid polypeptides; cells comprising such polynucleotides; and methods of producing such chimeric and hybrid polypeptides using such cells. 1121-. (canceled)122. A method of treating hemophilia B in a human subject in need thereof , comprising administering to the subject a dose of at least about 10 IU/kg of a chimeric Factor IX (FIX) polypeptide , comprising FIX and an FcRn binding partner (FcRn BP) , at a dosing interval of a week or longer.123. The method of claim 122 , wherein the subject exhibits a plasma FIX activity above 1 IU/dL during the dosing interval.124. The method of claim 122 , wherein the subject exhibits a baseline-subtracted plasma FIX activity trough level of at least 1 IU/dL claim 122 , and wherein the baseline is the lowest measured plasma FIX level in the subject prior to administering the first dose of the chimeric FIX polypeptide.125. The method of claim 122 , wherein the long-acting FIX polypeptide exhibits one or more characteristics selected from the group consisting of:a. an incremental recovery (K-Value) (activity; observed) greater than 0.75 IU/dL per IU/kg;b. an incremental recovery (K value) (activity; observed) of about 0.62-1.17 IU/dL per IU/kg;c. a clearance (CL) (activity) of about 1.84-4.58 mL/hour/kg;d. a clearance (CL) (activity) of about 3.36±0.93 mL/hour/kg;e. a mean residence time (MRT) (activity) of about 53.1-85.8 hours;f. a mean residence time (MRT) (activity) of about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, or about 90 hours;g. a MRT (activity) of at least about 68.05±11.16 hours;h. a t1/2beta (activity) of about 40-67.4 hours;i. a t1/2beta (activity) of at least about 40, about 45, about 50, about 55, about 60, about 65, about 70, or about 75 hours;j. a t1/2beta (activity) of at least 75 ...

FACTOR IX FUSION PROTEINS AND METHODS OF MAKING AND USING SAME

The present disclosure provides Factor IX (FIX) fusion proteins comprising at least one heterologous moiety, such as an XTEN. The present disclosure further discloses methods of making and using the FIX fusion proteins. 1. A method of administering a Factor IX (FIX) fusion protein to a subject in need thereof , comprising subcutaneously administering to the subject a FIX fusion protein , wherein(a) the FIX fusion protein comprises a FIX polypeptide and at least a first XTEN, which is inserted within the FIX polypeptide at an insertion site corresponding to an amino acid selected from the group consisting of amino acid 103 of SEQ ID NO: 2, amino acid 105 of SEQ ID NO: 2, amino acid 142 of SEQ ID NO: 2, amino acid 149 of SEQ ID NO: 2, amino acid 162 of SEQ ID NO: 2, amino acid 166 of SEQ ID NO: 2, amino acid 174 of SEQ ID NO: 2, amino acid 224 of SEQ ID NO: 2, amino acid 226 of SEQ ID NO: 2, amino acid 228 of SEQ ID NO: 2, amino acid 413 of SEQ ID NO: 2, and any combination thereof, and(b) wherein following the administration, the FIX fusion protein exhibits a plasma activity of from about 5% to about 30% in the subject.2. A method of administering a Factor IX (FIX) fusion protein to a subject in need thereof , comprising subcutaneously administering to the subject a FIX fusion protein comprising a FIX polypeptide and a first Fc domain , wherein the FIX fusion protein comprises an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: 229 , and wherein following the administration , the FIX fusion protein exhibits a plasma activity of from about 5% to about 30% in the subject.3. The method of claim 1 , wherein the FIX fusion protein is administered at a dose of about 50 IU/kg to about 400 IU/kg.4. The method of claim 1 , wherein the FIX fusion protein exhibits a plasma activity peak value of about 10% to about 30%.5. The method of claim 1 , wherein the FIX fusion protein exhibits a plasma activity trough value of about 5% to about 10%.6. The ...

Factor IX Polypeptides and Methods of Use Thereof

The present invention provides methods of administering Factor IX; methods of administering chimeric and hybrid polypeptides comprising Factor IX; polynucleotides encoding such chimeric and hybrid polypeptides; cells comprising such polynucleotides; and methods of producing such chimeric and hybrid polypeptides using such cells.

Factor IX Polypeptides and Methods of Use Thereof

The present invention provides methods of administering Factor IX; methods of administering chimeric and hybrid polypeptides comprising Factor IX; polynucleotides encoding such chimeric and hybrid polypeptides; cells comprising such polynucleotides; and methods of producing such chimeric and hybrid polypeptides using such cells. 1121-. (canceled)122. A method of prophylactic treatment of hemophilia B in a human subject in need thereof , comprising intravenously administering to the subject multiple doses of about 25 IU/kg to about 50 IU/kg of a chimeric polypeptide comprising human Factor IX (FIX) and a FcRn binding partner (FcRn BP) at a dosing interval of about 7 days between doses , wherein the FcRn BP is a human Fc or a human albumin , wherein the subject exhibits a baseline-subtracted plasma FIX activity trough level of at least 1 IU/dL , and wherein the baseline is the lowest measured plasma Factor IX level in the subject prior to administering the first dose of the chimeric polypeptide.123. The method of claim 122 , wherein the dosing interval is 7 days.124. The method of claim 122 , wherein each of the multiple doses is 25 IU/kg to 40 IU/kg.125. The method of claim 124 , wherein the dosing interval is 7 days.126. The method of claim 122 , wherein each of the multiple doses is 25 IU/kg claim 122 , 30 IU/kg claim 122 , 35 IU/kg claim 122 , 40 IU/kg claim 122 , 45 IU/kg claim 122 , or 50 IU/kg.127. The method of claim 122 , wherein each of the multiple doses is 25 IU/kg.128. The method of claim 122 , wherein each of the multiple doses is 30 IU/kg.129. The method of claim 122 , wherein each of the multiple doses is 35 IU/kg.130. The method of claim 122 , wherein each of the multiple doses is 40 IU/kg.131. The method of claim 122 , wherein each of the multiple doses is 45 IU/kg.132. The method of claim 122 , wherein each of the multiple doses is 50 IU/kg.133. The method of claim 122 , wherein the human FIX comprises amino acids 1 to 415 of SEQ ID NO: 2.134. The ...

CHIMERIC CLOTTING FACTORS

The invention provides chimeric clotting factors comprising an activatable clotting factor and an enhancer moiety. The activatable clotting factor allows the chimeric clotting factor to be activated at the site of coagulation. The enhancer moiety can additionally improve procoagulation activities of the chimeric clotting factors. The chimeric clotting factors can further be improved by fusion to a half-life extender, which improves a pharmacokinetics property of the chimeric clotting factor. The invention also includes methods of making and methods of using these chimeric clotting factors. 1. A chimeric protein comprising (i) an activatable clotting factor (Ac) , (ii) an enhancer moiety (Em) , and (iii) an optional linker moiety (L or L1) between the activatable clotting factor and an enhancer moiety.2. (canceled)3. The chimeric protein according to claim 1 , comprising a structure represented by formula Ac-L-Em or Em-L-Ac claim 1 , wherein Ac comprises the activatable clotting factor; wherein L comprises the optional linker moiety; wherein Em comprises the enhancer moiety claim 1 , wherein the optional linker moiety comprises a gly/ser peptide claim 1 , and optionally wherein the enhancer moiety comprises a clotting cofactor claim 1 , a procoaulant peptide claim 1 , or an antigen-binding moiety.4. The chimeric protein according to claim 3 , wherein the activatable clotting factor comprises a clotting factor zymogen comprising a heavy chain (HC) and a light chain (LC) and a protease-cleavage site inserted between the HC and the LC claim 3 , and optionally wherein the clotting factor zymogen is a FVII protein or a FX protein.58-. (canceled)9. The chimeric protein according to claim 4 , wherein:(a) the clotting factor zymogen comprises a FVII protein, and the clotting cofactor comprises a Tissue Factor protein; or(b) the clotting factor zymogen comprises a FX protein, and the clotting cofactor comprises a FVa protein.1011-. (canceled)12. The chimeric protein according ...

Factor IX Polypeptides and Methods of Use Thereof

The present invention provides methods of administering Factor IX; methods of administering chimeric and hybrid polypeptides comprising Factor IX; chimeric and hybrid polypeptides comprising Factor IX; polynucleotides encoding such chimeric and hybrid polypeptides; cells comprising such polynucleotides; and methods of producing such chimeric and hybrid polypeptides using such cells. 1121-. (canceled)122. A method of treating hemophilia B in a human subject in need thereof , comprising administering to the subject multiple doses of about 40 IU/kg to about 110 IU/kg of a long-acting Factor IX (“FIX”) polypeptide at a dosing interval of about 7 clays to about 14 clays between two doses , wherein the long-acting FIX polypeptide comprises FIX and an FcRn binding partner (FeRn BP).123. The method claim 122 , wherein each of the multiple doses is from about 40 IU/kg to about 50 IU/kg.124. The method claim 122 , wherein each of the multiple doses is from about 70 IU/kg to about 80 IU/kg.125. The method of claim 123 , wherein each of the multiple doses is from about 90 IU/kg to about 100 IU/kg.126. The method of claim 122 , wherein each of the multiple doses is 40 IU/kg to 50 IU/kg claim 122 , 50 IU/kg to 60 IU/kg claim 122 , 60 IU/kg to 70 IU/kg claim 122 , 70 IU/kg to 80 IU/kg claim 122 , 80 IU/kg to 90 IU/kg claim 122 , 90 IU/kg to 100 IU/kg claim 122 , or 100 IU/kg to 110 UI/kg.127. The method of claim 122 , wherein each of the multiple doses is 40 IU/kg claim 122 , 45 IU/kg claim 122 , 50 IU/kg claim 122 , 55 IU/kg claim 122 , 60 IU/kg claim 122 , 65 IU/kg claim 122 , 70 IU/kg claim 122 , 75 IU/kg claim 122 , 80 IU/kg claim 122 , 85 IU/kg claim 122 , 90 IU/kg claim 122 , 95 IU/kg claim 122 , 100 IU/kg claim 122 , 105 IU/kg claim 122 , or 110 IU/kg.128. The method of claim 122 , wherein the dosing interval is 7 days to 10 days.129. The method of claim 122 , wherein the dosing interval is 10 days to 14 days.130. The method of claim 122 , wherein each of the multiple doses ...

CLOTTING FACTOR-FC CHIMERIC PROTEINS TO TREAT HEMOPHILIA

The invention relates to a chimeric protein comprising at least one clotting factor and at least a portion of an immunoglobulin constant region. The invention relates to a method of treating a hemostatic disorder comprising administering a therapeutically effective amount of a chimeric protein wherein the chimeric protein comprises at least one clotting factor and at least a portion of an immunoglobulin constant region. 158.-. (canceled)59. A chimeric protein comprising a first polypeptide and a second polypeptide , wherein the first polypeptide comprises(i) a clotting factor, which is factor VIII, factor IX, factor XI, factor XII, fibrinogen, prothrombin, factor V, factor VII, factor VIIa, factor X, or factor XIII, and(ii) at least a portion of an immunoglobulin constant region fused to the clotting factor, which is a neonatal Fc Receptor (FcRn) binding partner, andthe second polypeptide comprises at least a portion of an immunoglobulin constant region,which is a FcRn binding partner, without the clotting factor of the first polypeptide and without an immunoglobulin variable domain, andwherein the first polypeptide and the second polypeptide are linked.60. The chimeric protein of claim 59 , wherein the clotting factor is fused to the portion of an immunoglobulin constant region by a linker.61. The chimeric protein of claim 60 , wherein the linker comprises about 1 to about 20 amino acids.62. The chimeric protein of claim 59 , wherein the portion of an immunoglobulin constant region of the first polypeptide is an Fc fragment.63. The chimeric protein of claim 62 , wherein the portion of an immunoglobulin constant region of the second polypeptide is an Fc fragment.64. The chimeric protein of claim 59 , wherein the portion of an immunoglobulin constant region of the first polypeptide and the portion of an immunoglobulin constant region of the second polypeptide are identical.65. The chimeric protein of claim 59 , wherein the clotting factor is full-length factor VIII ...

Methods of Using FIX Polypeptides

The present invention provides methods of administering long-acting Factor IX; methods of administering long-acting, chimeric and hybrid polypeptides comprising Factor IX; and methods of producing such chimeric and hybrid polypeptides using cells. 1. A method of administering a long-acting Factor IX (FIX) polypeptide to a human subject in need thereof , comprising administering to the subject a dose of about 10 IU/kg to about 200 IU/kg of the long-acting FIX polypeptide at a dosing interval of about once a week or longer.2. The method of claim 1 , wherein the subject is in need of treatment of a bleeding disorder.3. The method of or claim 1 , wherein the dose of the long-acting FIX polypeptide is about 10 IU/kg to about 50 IU/kg claim 1 , about 10 IU/kg to about 100 IU/kg claim 1 , about 25 IU/kg to about 50 IU/kg claim 1 , about 25 IU/kg to about 75 IU/kg claim 1 , about 25 IU/kg to about 100 IU/kg claim 1 , about 25 IU/kg to about 125 IU/kg claim 1 , about 25 IU/kg to about 150 IU/kg claim 1 , about 50 IU/kg to about 100 IU/kg claim 1 , about 50 IU/kg to about 150 IU/kg claim 1 , about 100 IU/kg to about 150 IU/kg claim 1 , about 150 IU/kg to about 200 IU/kg claim 1 , or any combinations thereof.4. The method of any one of to claim 1 , wherein the dose of the long-acting FIX polypeptide is for prophylaxis of one or more bleeding episodes.5. The method of claim 4 , wherein the dose is about 50 IU/kg.6. The method of or claim 4 , wherein an annualized bleeding rate of the bleeding episodes is less than 2 claim 4 , less than 3 claim 4 , less than 4 claim 4 , less than 5 claim 4 , less than 6 claim 4 , less than 7 claim 4 , less than 8 claim 4 , less than 9 claim 4 , or less than 10.7. The method of any one of to claim 4 , wherein the dose of the long-acting FIX polypeptide is for individualized interval prophylaxis of one or more bleeding episodes.8. The method of claim 7 , wherein the dose of the long-acting FIX polypeptide is about 100 IU/kg.9. The method of or ...

Factor IX Polypeptides and Methods of Use Thereof

The present invention provides methods of administering Factor IX; methods of administering chimeric and hybrid polypeptides comprising Factor IX; polynucleotides encoding such chimeric and hybrid polypeptides; cells comprising such polynucleotides; and methods of producing such chimeric and hybrid polypeptides using such cells. 1. A method of controlling or preventing a bleeding episode in a human subject in need thereof , comprising administering to the subject a dose of at least about 25 IU/kg of a chimeric polypeptide comprising Factor IX and a FcRn binding partner (FcRn BP) at about a once weekly or longer dosing interval.2. A method of prophylactic treatment of hemophilia in a human subject in need thereof , comprising administering to the subject a dose of at least about 25 IU/kg of a chimeric polypeptide comprising Factor IX and a FcRn binding partner (FcRn BP) at about a once weekly or longer dosing interval.39-. (canceled)10. The method of claim 1 , wherein said chimeric polypeptide exhibits one or more pharmacokinetic parameters claim 1 , in said patient population or in said subject claim 1 , selected from the group consisting of:a mean clearance (CL) (activity) in said patient population of about 3.36±0.93 mL/hour/kg;a mean clearance (CL) (activity) in said patient population of about 3.0-3.72, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, or 3.72 mL/hour/kg;a mean clearance (CL) (activity) in said patient population that is about 2.5 fold lower than the clearance of a polypeptide comprising said Factor IX without said FcRn BP;a clearance (CL) (activity) in said subject of about 1.84-4.58 mL/hour/kga mean residence time (MRT) (activity) in said patient population of at least about 68.05±11.16 hours;a mean MRT (activity) in said patient population of about 60-78, 60, 62, 64, 66, 68, 70, 72, 74, 76, or 78 hours;a mean MRT (activity) in said patent population that is about 3 fold longer than the mean MRT of a polypeptide comprising said Factor IX without said ...

FACTOR VIII COMPLEX WITH XTEN AND VON WILLEBRAND FACTOR PROTEIN, AND USES THEREOF

The present invention includes a chimeric protein comprising a VWF protein with D′ domain and D3 domain of VWF, one or more XTEN sequence, and a FVIII protein, wherein the VWF fragment, the XTEN sequence, or the FVIII protein are linked to or associated with each other. The chimeric protein can further comprise one or more Ig constant region or a portion thereof (e.g., an Fc region). A polypeptide chain of a VWF fragment is associated with a FVIII polypeptide chain linked to an XTEN sequence. The VWF fragment polypeptide chain can prevent or inhibit binding of endogenous VWF to FVIII protein linked to the XTEN sequence. By preventing or inhibiting binding of endogenous VWF to FVIII protein, VWF fragment can extend half-life of chimeric protein comprising FVIII protein. The invention includes nucleotides, vectors, host cells, use of VWF fragment, or chimeric proteins. 1. A chimeric protein comprising (i) a von Willebrand Factor (VWF) fragment comprising a D′ domain and a D3 domain of VWF , (ii) an extended length polypeptide (XTEN) sequence , and (iii) a factor VIII (FVIII) protein , wherein the VWF fragment and the XTEN sequence are linked by an optional linker , and wherein , the VWF fragment or the XTEN sequence is linked to or associated with the FVIII protein.24-. (canceled)5. The chimeric protein of claim 1 , which comprises(a) a single polypeptide chain comprising the VWF fragment, the XTEN sequence and the FVIII protein; or(b) two polypeptides comprising a first polypeptide and a second polypeptide, wherein the first polypeptide comprises the FVIII protein and the second polypeptide comprises the VWF fragment and the XTEN sequence.6. (canceled)7. The chimeric protein of claim 1 , further comprising (iv) an immunoglobulin (Ig) constant region or a portion thereof linked to either the VWF fragment claim 1 , the XTEN sequence claim 1 , or the FVIII protein claim 1 , or any combination thereof.8. The chimeric protein of claim 1 , comprising a formula claim 1 , ...

RECOMBINANT FACTOR VIII PROTEINS

Provided are recombinant Factor VIII proteins, e.g., human Factor VIII proteins, in which one or more amino acids in at least one permissive loops or a3 domain are substituted or deleted, or replaced with heterologous moieties, while retaining the procoagulant activity of Factor VIII. 1. A recombinant FVIII protein comprising: a first polypeptide comprising Formula I: (A1)-a1-(A2)-a2-[B]; and a second polypeptide comprising Formula II: a3-(A3)-(C1);wherein the first polypeptide and the second polypeptide are fused or associated as a heterodimer;wherein a) A1 is an A1 domain of FVIII; b) A2 is an A2 domain of FVIII; c) [B] is optionally present and is a B domain of FVIII, or a fragment thereof; d) A3 is an A3 domain of FVIII; e) C1 is a C1 domain of FVIII; and f) a1, a2, and a3 are acidic spacer regions;wherein one or more amino acids in a permissive loop-1 region in the A1 domain (A1-1), a permissive loop-2 region in the A2 domain (A1-2), a permissive loop-1 region in the A2 domain (A2-1), a permissive loop-2 region in the A2 domain (A2-2), a permissive loop-1 region in the A3 domain (A3-1), a permissive loop-2 region in the A3 domain (A3-2), the a3 region, or any combinations thereof are substituted or deleted;wherein a heterologous moiety is inserted in at least one of A1-1, A1-2, A2-1, A2-2, A3-1, A3-2, or the a3 region; andwherein the recombinant FVIII protein exhibits procoagulant activity.24-. (canceled)5. The recombinant FVIII protein of claim 1 , wherein the first polypeptide and the second polypeptide form a single polypeptide chain comprising the formula (A1)-a1-(A2)-a2-[B]-[a3]-(A3)-(C1).67-. (canceled)8. The recombinant FVIII protein of claim 1 , wherein the one or more amino acids substituted or deleted are in a domain selected from the group consisting of:(i) A1-1, which corresponds to a region in native mature human FVIII from about amino acid 15 to about amino acid 45 or from about amino acid 18 to about amino acid 41 of SEQ ID NO:1;(ii) A1-2, which ...

Processable Single Chain Molecules and Polypeptides Made Using Same

The present invention features inter alia nucleic acid molecules which encode polypeptides comprising a single chain Fc region and the polypeptides they encode. The Fc moieties of these constructs are linked by a cleavable scFc linker which is adjacent to at least one enzymatic cleavage site, e.g., an intracellular processing site. The resulting processed molecules comprise two polypeptide chains and substantially lack the extraneous amino acid sequence found in single chain Fc linker molecule. Methods of making and using these dimeric molecules are also described. 1. A polypeptide , comprising (i) at least one biologically active moiety , (ii) an Fc region comprising at least two Fc moieties and (iii) a cleavable scFc (cscFc) linker interposed between the two Fc moieties , wherein the cscFc linker is linked to at least one enzymatic cleavage site which results in cleavage of the cscFc linker.237-. (canceled)38. A nucleic acid molecule encoding the polypeptide of .39. A vector comprising the nucleic acid molecule of .40. (canceled)41. A processed polypeptide comprising at least two amino acid chains which polypeptide is encoded by the nucleic acid molecule of .42. A host cell comprising the vector of claim 39 , wherein the host cell expresses an enzyme which cleaves the polypeptide linker.43. The host cell of claim 42 , wherein the enzyme is endogenous to the cell.44. The host cell of claim 42 , wherein the enzyme is exogenous to the cell.45. A method for producing a polypeptide comprising culturing the host cell of in culture such that a mature polypeptide comprising two amino acid chains is produced.4647-. (canceled)48. A composition comprising the nucleic acid molecule of and a pharmaceutically acceptable carrier.49. A method for treating or preventing a disease or disorder in a subject claim 48 , comprising administering the composition of .5052-. (canceled)54. The nucleic acid molecule of claim 53 , wherein P1 and P2 are recognized by different enzymes.55. The ...

Factor IX Polypeptides and Methods of Use Thereof

The present invention provides methods of administering Factor IX; methods of administering chimeric and hybrid polypeptides comprising Factor IX; chimeric and hybrid polypeptides comprising Factor IX; polynucleotides encoding such chimeric and hybrid polypeptides; cells comprising such polynucleotides; and methods of producing such chimeric and hybrid polypeptides using such cells. 1121-. (canceled)122. A method of treating hemophilia B in a human subject in need thereof , comprising administering to the subject multiple doses of about 60 IU/kg to about 180 IU/kg of a long-acting Factor IX (“FIX”) polypeptide at a dosing interval of about 9 days to 18 days between two doses , wherein the long-acting FIX polypeptide comprises FIX and an FcRn binding partner (FcRn BP).123. The method of claim 122 , wherein each of the multiple doses is 60 IU/kg to 70 IU/kg claim 122 , 70 IU/kg to 80 IU/kg claim 122 , 80 IU/kg to 90 IU/kg claim 122 , 90 IU/kg to 100 IU/kg claim 122 , 100 IU/kg to 110 IU/kg claim 122 , 110 IU/kg to 120 IU/kg claim 122 , 120 IU/kg to 130 IU/kg claim 122 , or 130 IU/kg to 140 IU/kg.124. The method of claim 122 , wherein each of the multiple doses is 70 IU/kg to 80 IU/kg.125. The method of claim 122 , wherein each of the multiple doses is 90 IU/kg to 100 IU/kg.126. The method of claim 122 , wherein each of the multiple doses is 100 IU/kg to 110 IU/kg.127. The method of claim 122 , wherein each of the multiple doses is 60 IU/kg claim 122 , 65 IU/kg claim 122 , 70 IU/kg claim 122 , 75 IU/kg claim 122 , 80 IU/kg claim 122 , 85 IU/kg claim 122 , 90 IU/kg claim 122 , 95 IU/kg claim 122 , 100 IU/kg claim 122 , 105 IU/kg 110 IU/kg claim 122 , 115 IU/kg claim 122 , 120 IU/kg claim 122 , 125 IU/kg claim 122 , 130 IU/kg claim 122 , 135 IU/kg claim 122 , 140 IU/kg claim 122 , 145 IU/kg claim 122 , 150 IU/kg claim 122 , 155 IU/kg claim 122 , 160 IU/kg claim 122 , 165 IU/kg claim 122 , 170 IU/kg claim 122 , 175 IU/kg claim 122 , or 180 IU/kg.128. The method of claim ...

CHIMERIC CLOTTING FACTORS

The invention provides chimeric clotting factors comprising an activatable clotting factor and an enhancer moiety. The activatable clotting factor allows the chimeric clotting factor to be activated at the site of coagulation. The enhancer moiety can additionally improve procoagulation activities of the chimeric clotting factors. The chimeric clotting factors can further be improved by fusion to a half-life extender, which improves a pharmacokinetics property of the chimeric clotting factor. The invention also includes methods of making and methods of using these chimeric clotting factors. 1. A chimeric protein comprising (i) an activatable clotting factor (Ac) , (ii) an enhancer moiety (Em) , and (iii) an optionally linker moiety (L or L1) between the activatable clotting factor and an enhancer moiety.2. (canceled)3. The chimeric protein according to claim 1 , comprising a structure represented by formula Ac-L-Em or Em-L-Ac claim 1 , wherein Ac comprises the activatable clotting factor; wherein L comprises the optional linker moiety; and wherein Em comprises the enhancer moiety.4. The chimeric protein according to claim 1 , wherein the activatable clotting factor comprises a clotting factor zymogen comprising a heavy chain (HC) and a light chain (LC) and a protease-cleavage site inserted between the HC and the LC.5. The chimeric protein according to claim 1 , wherein the enhancer moiety comprises a clotting cofactor claim 1 , a procoagulant peptide claim 1 , or an antigen-binding moiety. 6 claim 1 , (Currently Amended) The chimeric protein according to claim 1 , wherein the clotting factor zymogen is a FVII protein or a FX protein.7. (canceled)8. (canceled)9. The chimeric protein according to claim 4 , wherein:(a) the clotting factor zymogen comprises a FVII protein, and the clotting cofactor comprises a Tissue Factor protein; or (b) the clotting factor zymogen comprises a FX protein, and the clotting cofactor comprises a FVa protein.10. (canceled)11. (canceled)12. ...

Optimized Factor VIII Gene

The present invention provides codon optimized Factor VIII sequences, vectors and host cells comprising codon optimized Factor VIII sequences, polypeptides encoded by codon optimized Factor VIII sequences, and methods of producing such polypeptides. 1. An isolated nucleic acid molecule comprising a nucleotide sequence at least 85% identical to SEQ ID NO:1 , wherein the nucleotide sequence encodes a polypeptide with Factor VIII activity.2. The isolated nucleic acid molecule of claim 1 , wherein the nucleotide sequence is at least 90% identical to SEQ ID NO:1.3. The isolated nucleic acid molecule of claim 1 , wherein the nucleotide sequence is at least 95% claim 1 , at least 96% claim 1 , at least 97% claim 1 , at least 98% claim 1 , at least 99% claim 1 , or 100% identical to SEQ ID NO:1.4. The isolated nucleic acid molecule of claim 1 , wherein the nucleotide sequence comprises SEQ ID NO:1.5. An isolated nucleic acid molecule comprising a nucleotide sequence at least 95% claim 1 , at least 96% claim 1 , at least 97% claim 1 , at least 98% claim 1 , at least 99% claim 1 , or 100% identical to SEQ ID NO:2 claim 1 , wherein the nucleotide sequence encodes a polypeptide with Factor VIII activity.6. The isolated nucleic acid molecule of claim 5 , wherein the nucleotide sequence comprises SEQ ID NO:2.76. The isolated nucleic acid molecule of any one of - claims 1 , wherein the human codon adaptation index is increased relative to SEQ ID NO:3.8. The isolated nucleic acid molecule of claim 7 , wherein the human codon adaptation index is at least about 0.75 claim 7 , at least about 0.76 claim 7 , at least about 0.77 claim 7 , at least about 0.78 claim 7 , at least about 0.79 claim 7 , or at least about 0.80.9. The isolated nucleic acid molecule of claim 7 , wherein the human codon adaptation index is at least about 0.80 claim 7 , at least about 0.81 claim 7 , at least about 0.82 claim 7 , at least about 0.83 claim 7 , at least about 0.84 claim 7 , at least about 0.85 claim 7 ...

FACTOR IX FUSION PROTEINS AND METHODS OF MAKING AND USING SAME

The present invention provides Factor IX (FIX) fusion proteins comprising at least one heterologous moiety, such as an XTEN. The present invention further discloses methods of making and using the FIX fusion proteins. 1. A Factor IX (FIX) fusion protein comprising a FIX polypeptide and at least one XTEN which is inserted within the FIX polypeptide at an insertion site corresponding to an amino acid selected from the group consisting of amino acid 103 of SEQ ID NO: 2 , amino acid 105 of SEQ ID NO: 2 , amino acid 142 of SEQ ID NO: 2 , amino acid 149 of SEQ ID NO: 2 , amino acid 162 of SEQ ID NO: 2 , amino acid 166 of SEQ ID NO: 2 , amino acid 174 of SEQ ID NO: 2 , amino acid 224 of SEQ ID NO: 2 , amino acid 226 of SEQ ID NO: 2 , amino acid 228 of SEQ ID NO: 2 , amino acid 413 of SEQ ID NO: 2 , and any combination thereof , and wherein the FIX fusion protein exhibits procoagulant activity.2. The FIX fusion protein of claim 1 , wherein the insertion site corresponds to an amino acid selected from the group consisting of amino acid 149 of SEQ ID NO: 2 claim 1 , amino acid 162 of SEQ ID NO: 2 claim 1 , amino acid 166 of SEQ ID NO: 2 claim 1 , amino acid 174 of SEQ ID NO: 2 and any combination thereof.3. The FIX fusion protein of claim 1 , wherein the XTEN comprises at least about 36 amino acids.4. The FIX fusion protein of claim 1 , which further comprises a second XTEN.5. The FIX fusion protein of claim 4 , wherein the XTEN is inserted within the FIX polypeptide at an insertion site corresponding to amino acid 166 of SEQ ID NO: 2 claim 4 , and wherein the second XTEN is fused to the C-terminus of the FIX polypeptide.6. The FIX fusion protein of claim 1 , further comprising an Fc domain.7. The FIX fusion protein of claim 6 , comprising a second Fc domain.8. The FIX fusion protein of claim 7 , which comprises two polypeptide chains claim 7 , wherein the first polypeptide chain comprises the FIX polypeptide fused to the Fc domain claim 7 , and the second polypeptide chain ...

Factor viii chimeric and hybrid polypeptides, and methods of use thereof

The present invention provides methods of administering Factor VIII (processed FVIII, single chain FVIII, or a combination thereof); methods of administering chimeric and hybrid polypeptides comprising Factor VIII; chimeric and hybrid polypeptides comprising Factor VIII; polynucleotides encoding such chimeric and hybrid polypeptides; cells comprising such polynucleotides; and methods of producing such chimeric and hybrid polypeptides using such cells.

Procoagulant compounds

The present disclosure provides protease-activatable procoagulant compounds comprising a procoagulant polypeptide, e.g., a procoagulant peptide and/or clotting factor, and a linker comprising a protease-cleavable substrate (e.g., a synthetic thrombin substrate) and a self-immolative spacer (e.g., p-amino benzyl carbamate). Upon cleavage of the protease-cleavable substrate by a protease (e.g., thrombin), the self-immolative spacer cleaves itself from the procoagulant polypeptide such that the polypeptide is in an underivatized and active form. Also provided are pharmaceutical compositions, methods for treating bleeding disorders using the disclosed compounds, methods of enhancing in vivo efficacy of procoagulant polypeptides, methods of increasing the efficacy of proteolytic cleavage of compounds comprising procoagulant polypeptides, methods of activating procoagulant polypeptides, and methods of releasing a procoagulant polypeptide from a heterologous moiety such as PEG.

Chimeric clotting factors

The invention provides chimeric clotting factors comprising an activatable clotting factor and an enhancer moiety. The activatable clotting factor allows the chimeric clotting factor to be activated at the site of coagulation. The enhancer moiety can additionally improve procoagulation activities of the chimeric clotting factors. The chimeric clotting factors can further be improved by fusion to a half-life extender, which improves a pharmacokinetics property of the chimeric clotting factor. The invention also includes methods of making and methods of using these chimeric clotting factors.

Procoagulant compounds

The present disclosure provides protease-activatable procoagulant compounds comprising a procoagulant polypeptide, e.g., a procoagulant peptide and/or clotting factor, and a linker comprising a protease-cleavable substrate (e.g., a synthetic thrombin substrate) and a self-immolative spacer (e.g., p-amino benzyl carbamate). Upon cleavage of the protease-cleavable substrate by a protease (e.g., thrombin), the self-immolative spacer cleaves itself from the procoagulant polypeptide such that the polypeptide is in an underivatized and active form. Also provided are pharmaceutical compositions, methods for treating bleeding disorders using the disclosed compounds, methods of enhancing in vivo efficacy of procoagulant polypeptides, methods of increasing the efficacy of proteolytic cleavage of compounds comprising procoagulant polypeptides, methods of activating procoagulant polypeptides, and methods of releasing a procoagulant polypeptide from a heterologous moiety such as PEG.

Chimeric clotting factors

The invention provides chimeric clotting factors comprising an activatable clotting factor and an enhancer moiety. The activatable clotting factor allows the chimeric clotting factor to be activated at the site of coagulation. The enhancer moiety can additionally improve procoagulation activities of the chimeric clotting factors. The chimeric clotting factors can further be improved by fusion to a half-life extender, which improves a pharmacokinetics property of the chimeric clotting factor. The invention also includes methods of making and methods of using these chimeric clotting factors.

Procoagulant compounds

The present disclosure provides protease-activatable procoagulant compounds comprising a procoagulant polypeptide, e.g., a procoagulant peptide and/or clotting factor, and a linker comprising a protease-cleavable substrate ( e.g ., a synthetic thrombin substrate) and a self-immolative spacer ( e.g ., p-amino benzyl carbamate). Upon cleavage of the protease-cleavable substrate by a protease ( e.g ., thrombin), the self-immolative spacer cleaves itself from the procoagulant polypeptide such that the polypeptide is in an underivatized and active form. Also provided are pharmaceutical compositions, methods for treating bleeding disorders using the disclosed compounds, methods of enhancing in vivo efficacy of procoagulant polypeptides, methods of increasing the efficacy of proteolytic cleavage of compounds comprising procoagulant polypeptides, methods of activating procoagulant polypeptides, and methods of releasing a procoagulant polypeptide from a heterologous moiety such as PEG.

Procoagulant compounds

Bispecific antibodies binding to coagulation factor ix and coagulation factor x

The present disclosure provides antibodies that selectively binds to specific forms of clotting factors, in particular, antibodies that specifically binds to activated factor IX (FIXa) wherein the anti-FIXa antibody or an antigen binding portion thereof preferentially binds to FIXa in the presence of FIXa and factor IX zymogen (FIXz), and antibodies that specifically bind to factor X zymogen (FXz) wherein the anti-FXz antibody or antigen binding portion thereof preferentially binds to FXz in the presence of FXz and activated factor X (FXa). Also provided are bispecific molecules (e.g., antibodies) comprising, e.g., an anti-FIXa antibody or antigen binding portion thereof and/or an anti-FXz antibody or antigen binding portion thereof. The disclosure also provides compositions encoding the disclosed antibodies and bispecific molecules, vectors, cells, pharmaceutical and diagnostic compositions, kits, methods of manufacture, methods of use, and immunoconjugates.

Immunoglobulin chimeric monomer-dimer hybrids

The invention relates to a chimeric monomer-dimer hybrid protein wherein the protein comprises a first and a second polypeptide chain, the first polypeptide chain comprising at least a portion of an immunoglobulin constant region and a biologically active molecule, and the second polypeptide chain comprising at least a portion of an immunoglobulin constant region without the biologically active molecule of the first chain. The invention also relates to methods of using and methods of making the chimeric monomer-dimer hybrid protein of the invention.

Processable single chain molecules and polypeptides made using same

The present invention features inter alia nucleic acid molecules which encode polypeptides comprising a single chain Fc region and the polypeptides they encode. The Fc moieties of these constructs are linked by a cleavable scFc linker which is adjacent to at least one enzymatic cleavage site, e.g., an intracellular processing site. The resulting processed molecules comprise two polypeptide chains and substantially lack the extraneous amino acid sequence found in single chain Fc linker molecule. Methods of making and using these dimeric molecules are also described.

Immunoglobulin chimeric monomer-dimer hybrids

Clotting factor-fc chimeric proteins to treat hemophilia

The invention relates to a chimeric protein comprising at least one clotting factor and at least a portion of an immunoglobulin constant region. The invention relates to a method of treating a hemostatic disorder comprising administering a therapeutically effective amount of a chimeric protein wherein the chimeric protein comprises at least one clotting factor and at least a portion of an immunoglobulin constant region.

Methods for chemically synthesizing immunoglobulin chimeric proteins

The invention provides methods of chemically synthesizing chimeric proteins comprising at least a portion of an immunoglobulin constant region and a biologically active molecule.

Clotting Factor-Fc Chimeric Proteins to Treat Hemophilia

The invention relates to a chimeric protein comprising at least one clotting factor and at least a portion of an immunoglobulin constant region. The invention relates to a method of treating a hemostatic disorder comprising administering a therapeutically effective amount of a chimeric protein wherein the chimeric protein comprises at least one clotting factor and at least a portion of an immunoglobulin constant region.

Immunoglobulin chimeric monomer-dimer hybrids

The invention relates to a chimeric monomer-dimer hybrid protein wherein the protein comprises a first and a second polypeptide chain, the first polypeptide chain comprising at least a portion of an immunoglobulin constant region and a biologically active molecule, and the second polypeptide chain comprising at least a portion of an immunoglobulin constant region without the biologically active molecule of the first chain. The invention also relates to methods of using and methods of making the chimeric monomer-dimer hybrid protein of the invention.

Clotting factor-fc chimeric proteins to treat hemophilia

Clotting Factor-FC Chimeric Proteins to Treat Hemophilia

The invention relates to a chimeric protein comprising at least one clotting factor and at least a portion of an immunoglobulin constant region. The invention relates to a method of treating a hemostatic disorder comprising administering a therapeutically effective amount of a chimeric protein wherein the chimeric protein comprises at least one clotting factor and at least a portion of an immunoglobulin constant region.

Clotting Factor-Fc Chimeric Proteins to Treat Hemophilia

The invention relates to a chimeric protein comprising at least one clotting factor and at least a portion of an immunoglobulin constant region. The invention relates to a method of treating a hemostatic disorder comprising administering a therapeutically effective amount of a chimeric protein wherein the chimeric protein comprises at least one clotting factor and at least a portion of an immunoglobulin constant region.

Immunoglobulin chimeric monomer-dimer hybrids

The invention relates to a chimeric monomer-dimer hybrid protein wherein the protein comprises a first and a second polypeptide chain, the first polypeptide chain comprising at least a portion of an immunoglobulin constant region and a biologically active molecule, and the second polypeptide chain comprising at least a portion of an immunoglobulin constant region without the biologically active molecule of the first chain. The invention also relates to methods of using and methods of making the chimeric monomer-dimer hybrid protein of the invention.

Immunoglobulin chimeric monomer-dimer hybrids

The invention relates to a chimeric monomer-dimer hybrid protein wherein the protein comprises a first and a second polypeptide chain, the first polypeptide chain comprising at least a portion of an immunoglobulin constant region and a biologically active molecule, and the second polypeptide chain comprising at least a portion of an immunoglobulin constant region without the biologically active molecule of the first chain. The invention also relates to methods of using and methods of making the chimeric monomer-dimer hybrid protein of the invention.

Clotting factor VII-Fc chimeric proteins for treatment of a hemostatic disorder

A pharmaceutical composition, for treatment of a hemostatic disorder, comprises a chimeric protein and a pharmaceutically acceptable carrier or excipient, wherein said chimeric protein comprises at least a portion of an immunoglobulin constant region which is an FeRn binding partner, a clotting factor selected from Factor VII, a Factor VII analog, Factor VIIa and a Factor VIIa analog, and optionally at least one linker.

Immunoglobulin chimeric monomer-dimer hybrids

The invention relates to a chimeric monomer-dimer hybrid protein that comprises a first and a second polypeptide chain, the first polypeptide chain comprising at least a portion of an immunoglobulin constant region and a biologically active molecule, and the second polypeptide chain comprising at least a portion of an immunoglobulin constant region without the biologically active molecule of the first polypeptide chain. The invention also relates to methods of using and methods of making the chimeric monomer-dimer hybrid protein of the invention.

Clotting factor-fc chimeric proteins to treat hemophilia

The invention relates to a chimeric protein comprising at least one clotting factor and at least a portion of an immunoglobulin constant region. The invention relates to a method of treating a hemostatic disorder comprising administering a therapeutically effective amount of a chimeric protein wherein the chimeric protein comprises at least one clotting factor and at least a portion of an immunoglobulin constant region.

Clotting factor-Fc chimeric proteins to treat hemophilia

The invention relates to a chimeric protein comprising at least one clotting factor and at least a portion of an immunoglobulin constant region. The invention relates to a method of treating a hemostatic disorder comprising administering a therapeutically effective amount of a chimeric protein wherein the chimeric protein comprises at least one clotting factor and at least a portion of an immunoglobulin constant region.

Immunoglobulin chimeric monomer-dimer hybrids

The invention relates to a chimeric monomer-dimer hybrid protein wherein the protein comprises a first and a second polypeptide chain, the first polypeptide chain comprising at least a portion of an immunoglobulin constant region and a biologically active molecule, and the second polypeptide chain comprising at least a portion of an immunoglobulin constant region without the biologically active molecule of the first chain. The invention also relates to methods of using and methods of making the chimeric monomer-dimer hybrid protein of the invention.

Immunoglobulin chimeric monomer-dimer hybrids

The invention relates to a chimeric monomer-dimer hybrid protein wherein the protein comprises a first and a second polypeptide chain, the first polypeptide chain comprising at least a portion of an immunoglobulin constant region and a biologically active molecule, and the second polypeptide chain comprising at least a portion of an immunoglobulin constant region without the biologically active molecule of the first chain. The invention also relates to methods of using and methods of making the chimeric monomer-dimer hybrid protein of the invention.

Clotting factor-Fc chimeric proteins to treat hemophilia

The invention relates to a chimeric protein comprising at least one clotting factor and at least a portion of an immunoglobulin constant region. The invention relates to a method of treating a hemostatic disorder comprising administering a therapeutically effective amount of a chimeric protein wherein the chimeric protein comprises at least one clotting factor and at least a portion of an immunoglobulin constant region.

Clotting factor-Fc chimeric proteins to treat hemophilia

Clotting factor VII-Fc chimeric proteins for treatment of a hemostatic disorder

A pharmaceutical composition, for treatment of a hemostatic disorder, comprises a chimeric protein and a pharmaceutically acceptable carrier or excipient, wherein said chimeric protein comprises at least a portion of an immunoglobulin constant region which is an FcRn binding partner, a clotting factor selected from Factor VII, a Factor VII analog, Factor VIIa and a Factor VIIa analog, and optionally at least one linker.

Clotting factor-fc chimeric proteins to treat hemophilia

A pharmaceutical composition, for treatment of a hemostatic disorder, comprises a chimeric protein and a pharmaceutically acceptable carrier or excipient, wherein said chimeric protein comprises at least a portion of an immunoglobulin constant region which is an FcRn binding partner, a clotting factor selected from Factor VII. a Factor VII analog, Factor VIIa and a Factor VIIa analog, and optionally at least one linker.

Factor viii-xten fusions and uses thereof

The present invention provides chimeric proteins comprising a Factor VIII (FVIII) polypeptide and XTEN polypeptides. The FVIII polypeptide can have a reduced affinity for von Willebrand Factor (VWF). XTEN polypeptides of appropriate sizes are inserted into the FVIII polypeptide at particular locations in order to extend the half-life of the FVIII polypeptide. The invention also includes nucleotides, vectors, host cells, and methods of using the chimeric proteins, e.g., to treat bleeding diseases and disorders.

factor ix fusion proteins and methods of manufacturing and using them

a presente invenção fornece proteínas de fusão do fator ix (fix) que compreendem pelo menos uma porção heteróloga, tal como um xten. a presente invenção descreve ainda métodos de fabricação e uso das proteínas de fusão fix. The present invention provides factor Ix (fix) fusion proteins comprising at least one heterologous moiety, such as an xten. The present invention further describes methods of making and using fixed fusion proteins.

Factor VIII complex with XTEN and von Willebrand Factor protein, and uses thereof

The present invention provides a chimeric protein comprising a VWF protein comprising the D′ domain and D3 domain of VWF, one or more XTEN sequence, and a FVIII protein, wherein the VWF fragment, the XTEN sequence, or the FVIII protein are linked to or associated with each other. The chimeric protein can further comprise one or more Ig constant region or a portion thereof (e.g., an Fc region). A polypeptide chain comprising a VWF fragment of the invention binds to or is associated with a polypeptide chain comprising a FVIII protein linked to an XTEN sequence and the polypeptide chain comprising the VWF fragment can prevent or inhibit binding of endogenous VWF to the FVIII protein linked to the XTEN sequence. By preventing or inhibiting binding of endogenous VWF to the FVIII protein, which is a half-life limiting factor for FVIII, the VWF fragment can induce extension of half-life of the chimeric protein comprising a FVIII protein. The invention also includes nucleotides, vectors, host cells, methods of using the VWF fragment, or the chimeric proteins.

Nucleic acid molecules and uses thereof

The present disclosure provides nucleic acid molecules comprising a first inverted terminal repeat (ITR), a second ITR, and a genetic cassette encoding a miRNA and/or a therapeutic protein. In certain embodiments, the therapeutic protein comprises a clotting factor, e.g., a FVIII polypeptide, a FIX polypeptide, or a fragment thereof. In some embodiments, the first ITR and/or the second ITR is an ITR of a non-adeno-associated virus (AAV). The present disclosure also provides methods of treating bleeding disorders such as hemophilia comprising administering to the subject the nucleic acid molecule or a polypeptide encoded thereby.

Optimized factor VIII gene

The present invention provides codon optimized Factor VIII sequences, vectors and host cells comprising codon optimized Factor VIII sequences, polypeptides encoded by codon optimized Factor VIII sequences, and methods of producing such polypeptides.

Nucleic acid molecules and uses thereof for non-viral gene therapy

The present disclosure provides nucleic acid molecules comprising a first inverted terminal repeat (ITR), a second ITR, and a genetic cassette encoding a target sequence. In some embodiments, the target sequence encodes a miRNA and/or a therapeutic protein. In certain embodiments, the therapeutic protein comprises a clotting factor, a growth factor, a hormone, a cytokine, an antibody, a fragment thereof, and a combination thereof. In some embodiments, the first ITR and/or the second ITR is an ITR of a non-adeno-associated virus (AAV). The present disclosure also provides methods of treating a metabolic disorder of the liver in a subject comprising administering to the subject the nucleic acid molecule or a polypeptide encoded thereby.

Factor IX fusion proteins and methods of making and using same

The present invention provides Factor IX (FIX) fusion proteins comprising at least one heterologous moiety, such as an XTEN. The present invention further discloses methods of making and using the FIX fusion proteins.

Optimized factor IX gene

The present invention provides codon optimized Factor IX sequences, vectors and host cells comprising codon optimized Factor IX sequences, polypeptides encoded by codon optimized Factor IX sequences, and methods of producing such polypeptides. The present invention also provides methods of treating bleeding disorders such as hemophilia comprising administering to the subject a codon optimized Factor IX nucleic acid sequence or the polypeptide encoded thereby.

Chimeric factor VIII polypeptides and uses thereof

The present invention provides a VWF fragment comprising the D′ domain and D3 domain of VWF, a chimeric protein comprising the VWF fragment and a heterologous moiety, or a chimeric protein comprising the VWF fragment and a FVIII protein and methods of using the same. A polypeptide chain comprising a VWF fragment of the invention binds to or is associated with a polypeptide chain comprising a FVIII protein and the polypeptide chain comprising the VWF fragment can prevent or inhibit binding of endogenous VWF to the FVIII protein. By preventing or inhibiting binding of endogenous VWF to the FVIII, which is a half-life limiting factor for FVIII, the VWF fragment can induce extension of half-life of the FVIII protein. The invention also includes nucleotides, vectors, host cells, methods of using the VWF fragment, or the chimeric proteins.

Factor viii complex with xten and von willebrand factor protein, and uses thereof

The present invention provides a chimeric protein comprising a VWF protein comprising the D' domain and D3 domain of VWF, one or more XTEN sequence, and a FVIII protein, wherein the VWF fragment, the XTEN sequence, or the FVIII protein are linked to or associated with each other. The chimeric protein can further comprise one or more Ig constant region or a portion thereof (e.g., an Fc region). A polypeptide chain comprising a VWF fragment of the invention binds to or is associated with a polypeptide chain comprising a FVIII protein linked to an XTEN sequence and the polypeptide chain comprising the VWF fragment can prevent or inhibit binding of endogenous VWF to the FVIII protein linked to the XTEN sequence. By preventing or inhibiting binding of endogenous VWF to the FVIII protein, which is a half-life limiting factor for FVIII, the VWF fragment can induce extension of half-life of the chimeric protein comprising a FVIII protein. The invention also includes nucleotides, vectors, host cells, methods of using the VWF fragment, or the chimeric proteins.

Factor VIII complex with XTEN and von Willebrand Factor protein, and uses thereof

The present invention provides a chimeric protein comprising a VWF protein comprising the D′ domain and D3 domain of VWF, one or more XTEN sequence, and a FVIII protein, wherein the VWF fragment, the XTEN sequence, or the FVIII protein are linked to or associated with each other. The chimeric protein can further comprise one or more Ig constant region or a portion thereof (e.g., an Fc region). A polypeptide chain comprising a VWF fragment of the invention binds to or is associated with a polypeptide chain comprising a FVIII protein linked to an XTEN sequence and the polypeptide chain comprising the VWF fragment can prevent or inhibit binding of endogenous VWF to the FVIII protein linked to the XTEN sequence. By preventing or inhibiting binding of endogenous VWF to the FVIII protein, which is a half-life limiting factor for FVIII, the VWF fragment can induce extension of half-life of the chimeric protein comprising a FVIII protein. The invention also includes nucleotides, vectors, host cells, methods of using the VWF fragment, or the chimeric proteins.

Optimized factor viii gene

The present invention provides codon optimized Factor VIII sequences, vectors and host cells comprising codon optimized Factor VIII sequences, polypeptides encoded by codon optimized Factor VIII sequences, and methods of producing such polypeptides. The present invention also provides methods of treating bleeding disorders such as hemophilia comprising administering to the subject a codon optimized Factor VIII nucleic acid sequence or the polypeptide encoded thereby.

Factor VIII-XTEN fusions and uses thereof

The present invention provides chimeric proteins comprising a Factor VIII (FVIII) polypeptide and XTEN polypeptides. The FVIII polypeptide can have a reduced affinity for von Willebrand Factor (VWF). XTEN polypeptides of appropriate sizes are inserted into the FVIII polypeptide at particular locations in order to extend the half-life of the FVIII polypeptide. The invention also includes nucleotides, vectors, host cells, and methods of using the chimeric proteins, e.g., to treat bleeding diseases and disorders.

Factor VIII chimeric proteins and uses thereof

The present invention provides a chimeric protein comprising a first polypeptide which comprises a FVIII protein and a first Ig constant region or a portion thereof and a second polypeptide which comprises a VWF protein comprising the D′ domain and D3 domain of VWF, a XTEN sequence having less than 288 amino acids in length, and a second Ig constant region or a portion thereof, wherein the first polypeptide and the second polypeptide are associated with each other. The invention also includes nucleotides, vectors, host cells, methods of using the chimeric proteins.

CHIMERIC FACTOR VIII POLYPEPTIDES AND USES THEREOF

The present invention provides a VWF fragment comprising the D′ domain and D3 domain of VWF, a chimeric protein comprising the VWF fragment and a heterologous moiety, or a chimeric protein comprising the VWF fragment and a FVIII protein and methods of using the same. A polypeptide chain comprising a VWF fragment of the invention binds to or is associated with a polypeptide chain comprising a FVIII protein and the polypeptide chain comprising the VWF fragment can prevent or inhibit binding of endogenous VWF to the FVIII protein. By preventing or inhibiting binding of endogenous VWF to the FVIII, which is a half-life limiting factor for FVIII, the VWF fragment can induce extension of half-life of the FVIII protein. The invention also includes nucleotides, vectors, host cells, methods of using the VWF fragment, or the chimeric proteins.

FACTOR VIII COMPLEX WITH XTEN AND VON WILLEBRAND FACTOR PROTEIN AND USES THEREOF

Optimized factor ix gene

Factor VIII chimeric proteins and uses thereof

The present invention provides a chimeric protein comprising a first polypeptide which comprises a FVIII protein and a first Ig constant region or a portion thereof and a second polypeptide which comprises a VWF protein comprising the D' domain and D3 domain of VWF, a XTEN sequence having less than 288 amino acids in length, and a second Ig constant region or a portion thereof, wherein the first polypeptide and the second polypeptide are associated with each other. The invention also includes nucleotides, vectors, host cells, methods of using the chimeric proteins.

Factor ix polypeptides and methods of use thereof

The present invention provides methods of administering Factor IX; methods of administering chimeric and hybrid polypeptides comprising Factor IX; chimeric and hybrid polypeptides comprising Factor IX; polynucleotides encoding such chimeric and hybrid polypeptides; cells comprising such polynucleotides; and methods of producing such chimeric and hybrid polypeptides using such cells.

Optimized factor ix gene

The present invention provides codon optimized Factor IX sequences, vectors and host cells comprising codon optimized Factor IX sequences, polypeptides encoded by codon optimized Factor IX sequences, and methods of producing such polypeptides. The present invention also provides methods of treating bleeding disorders such as hemophilia comprising administering to the subject a codon optimized Factor IX nucleic acid sequence or the polypeptide encoded thereby.

Imidazoline derivatives and their salts

ABSTRACT OF THE INVENTION Imidazoline compounds of formula (I): (I) wherein Ar is a phenyl or a mono-, di- or tri-substituted phenyl radical in which the substituents are alkyl, alkoxy, halogen, hydroxy, cyano, amino, trihalomethyl or nitro and in which any two adjacent carbon atoms of the phenyl ring may be joined by a carbon chain containing 3 or 4 carbon atoms; X1 is S, NR3 or NZ1; R9 and R2 are hydrogen or alkyl; R3 is alkyl or aryl; m is 0 or 1; and Z1 and Z2 are a group SOnR9 or a group in which X2 is O, S or NR5; R4 is alkyl, aryl, alkoxy, aryloxy, amino or amido; R5 is alkyl, aryl, alkoxy, aryloxy, alkylthio, arylthio, amino or amido; alkyl or alkoxy radicals containing from 1 to 4 carbon atoms each; or an acid addition salt of the imidazoline compound. Methods of preparing the compounds are provided, as are pesticidal formulations containing them. The compounds are active against pests, especially arthropods of the Order Acarina.