185results about "Factor VII" patented technology

The invention relates generally to compositions and methods for purifying the desired species from a mixture of desired heterodimer and contaminating homodimer immunoglobulin variants by modifying the isoelectric point(s) of the individual chains.

The invention relates generally to compositions and methods for purifying the desired species from a mixture of desired heterodimer and contaminating homodimer immunoglobulin variants by modifying the isoelectric point(s) of the individual chains.

The present invention relates to engineered liver-specific core promoters, synthetic promoters (which contains the engineered core promoters and enhancers), expression vectors (which contains the synthetic promoter), as well as methods of using the promoter or the expression vector thereof to address the need in the field, including treatment of various genetic diseases or conditions associated with the liver. In some embodiments, the liver-specific promoter includes continuous or discontinuous genome sequences from SERPINA1 genome.

To provide FVIII drugs with increased in vivo half-life while retaining sufficient biological activity, and prevention methods for hemophilia subjects.SOLUTION: The invention provides a conjugate comprising recombinant FVIII (rFVIII) and a zwitterionic polymer, where the polymer comprises one or more monomer units and where at least one monomer unit comprises a zwitterionic group. Optionally, the zwitterionic group comprises phosphorylcholine. In some conjugates, the polymer is covalently bonded to at least one of an amino group, a hydroxyl group, a sulfhydryl group and a carboxyl group of rFVIII.SELECTED DRAWING: Figure 2

Disclosed herein are nucleic acid compositions and methods of use. The nucleic acid compositions may have a therapeutic nucleic acid sequence operably linked to a nuclear targeting sequence that increases expression of the therapeutic nucleic acid in a cell by at least 1.25 fold; at least 80% sequence identity to SEQ ID NO: 6; or a first regulatory element comprising a promoter sequence operably linked to a hemoglobin subunit gamma intron (hBGi) sequence, and a second regulatory element comprising a woodchuck hepatitis posttranscriptional regulatory element (WPRE) sequence. The nucleic acid compositions with these features may enhance therapeutic nucleic acid transfection and expression. Also disclosed herein are transgenes optimized for gene therapy applications, including novel FVIII transgene sequences, in which the B domain may be non-naturally occurring the A1 and / or A3 domain may include at least one amino acid substitution.

Compositions and methods for treating Factor X deficiency are disclosed. The compositions and methods include the use of a a FVIII mimetic bispecific antibody that binds Factor IXa and Factor X.

This disclosure generally relates to super minimal transposon inverted repeat sequence (ITR) polynucleotides, compositions comprising the polynucleotides and methods of using compositions comprising the polynucleotides for the ex vivo and in vivo delivery of nucleic acids to cells, in particular, in vivo delivery of therapeutic genes to treat genetic disorders or diseases.

Among other aspects, the present invention relates to cell culture conditions for producing high molecular weight vWF, in particular, highly multimericWF with a high specific activity and ADAMTS13 with a high specific activity. The cell culture conditions of the present invention can include, for example, a cell culture medium with an increased copper concentration and / or cell culture supernatant with a low ammonium (NH4+) concentration. The present invention also provides methods for cultivating cells in the cell culture conditions to express high molecular weight vWF and rA13 having high specific activities.

Methods and assays for determining the activation level of the plasma kallikrein (pKal) system and the uses thereof for assessing the activity of pKal modulators on the pKal system.

Variants of coagulation factor VIII (FVIII) and expression cassettes encoding the FVIII variants thereof are described. A variant FVIII includes a glycoepitope of the FVIII protein including an N2118Q mutation. The N2118Q mutation can be combined with other mutations including a BDD-FVIII, N6, V3, RH, furin-cleavage site deletion. X10, K12, and / or F309S mutation to form additional FVIII variants. The FVIII variants with the N2118Q mutation and expression cassettes thereof can result in reduced immunogenicity of the resulting protein. When combined with other FVIII mutations, higher gene expression, increased secretion, increased stability, and higher FVIII functional activity can be achieved by the expressed FVIII variants. The variant FVIII and expression cassettes described here can be useful in protein replacement therapy and / or gene therapy for the treatment of hemophilia A.

The application describes ceDNA vectors having linear and continuous structure for delivery and expression of a transgene. ceDNA vectors comprise an expression cassette flanked by two ITR sequences, where the expression cassette encodes a transgene encoding FVIII protein. Some ceDNA vectors further comprise cis-regulatory elements, including regulatory switches. Further provided herein are methods and cell lines for reliable gene expression of FVIII protein in vitro, ex vivo and in vivo using the ceDNA vectors. Provided herein are method and compositions comprising ceDNA vectors useful for the expression of FVIII protein in a cell, tissue or subject, and methods of treatment of diseases with said ceDNA vectors expressing FVIII protein. Such FVIII protein can be expressed for treating disease, e.g., hemophilia A.

Compositions and regimens useful in treating Wilson's Disease are provided. The compositions include recombinant adeno-associated virus (rAAV) with a transthyretin enhancer and promoter driving expression of a human ATP7B.

Lentiviral vector (LV) formulations, and pharmaceutical compositions comprising such LV formulations, with improved stability and suitable for systemic administration are provided. Methods for treating disorders, especially blood disorders, using systemic administration of LV formulations are also provided.

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.

The invention provides a preparation method of a recombinant human blood coagulation factor VIII and a product of the recombinant human blood coagulation factor VIII. The preparation method comprises the step of culturing a mammalian cell strain for expressing the recombinant human blood coagulation factor VIII in a serum-free culture medium of a compound containing copper ions and methionine sulfoximine. By adopting the preparation method provided by the invention, the purity of the recombinant human blood coagulation factor VIII product can be further improved on the basis of keeping the high activity and high yield of the recombinant human blood coagulation factor VIII product obtained by culture.

The invention discloses a human blood coagulation factor VIII and a preparation method thereof, and the preparation method comprises the following steps: S1, carrying out refrigerated centrifugation on cryoprecipitate to obtain an extracting solution; s2, mixing the extracting solution and the heparin sodium solution to obtain a dissolving solution; s3, adjusting the pH value of the dissolving solution to 6.2-6.4 by using hydrochloric acid, and controlling the temperature to be 20-30 DEG C to obtain an acid precipitate; s4, mixing the acid precipitate with polyethylene glycol to obtain a polyethylene glycol precipitate; s5, adding the carboxylated ferroferric oxide nano-particles into the polyethylene glycol precipitate, incubating, and carrying out magnetic separation to obtain a first separation solution; s6, performing centrifugal separation on the first separation liquid to obtain a second separation liquid; s7, clarifying and filtering the second separation liquid, and adjusting the pH value to 6.4-7.4 to obtain a clarified liquid; and S8, adding a stabilizer and an inactivator into the clear liquid, and carrying out water bath heat preservation to obtain a human blood coagulation factor VIII stock solution. The method has the effect of improving the recovery activity and purity of the human blood coagulation factor VIII.

This disclosure generally relates to super minimal transposon inverted repeat sequence (ITR) polynucleotides, compositions comprising the polynucleotides and methods of using compositions comprising the polynucleotides for the ex vivo and in vivo delivery of nucleic acids to cells, in particular, in vivo delivery of therapeutic genes to treat genetic disorders or diseases.

The present disclosure provides pharmaceutical compositions of chimeric proteins comprising a factor VIII (FVIII) polypeptide and a von Willebrand factor (vWF) polypeptide. Pharmaceutical kits and methods of using the disclosed pharmaceutical compositions for treating hemophilia A are also disclosed.

The present invention refers to nucleotide sequences used for driving the expression of a therapeutic gene, preferably FVIII and / or its variants specifically in endothelial cells and / or hematopoietic, preferably myeloid cells. The sequences are useful for gene and / or cell therapy, preferably for treating hemophilia, more preferably type A hemophilia.

Lentiviral vector (LV) formulations, and pharmaceutical compositions comprising such LV formulations, with improved stability and suitable for systemic administration are provided. Methods for treating disorders, especially blood disorders, using systemic administration of LV formulations are also provided.

The present invention provides a chimeric protein comprising a first polypeptide comprising a FVIII protein and a first Ig constant region or a portion thereof; and a second polypeptide comprising a VWF protein comprising a D' domain and a D3 domain of VWF, an 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 one another. The present invention also includes nucleotides, vectors, host cells, methods of using the chimeric protein.

The present invention relates to the field of medicine and hematology, and provides compositions and methods for modulating Factor VIII function. In particular, the present invention provides novel factor VIII variants, compositions comprising the variants, nucleic acid molecules encoding the variants, expression vectors comprising the nucleic acid molecules, and host cells comprising the vectors. The present invention also provides methods of using the variants and / or compositions to modulate the coagulation cascade in a patient in need thereof for the treatment of diseases including, but not limited to, hemophilia, particularly hemophilia A.

Modified human factor VIII polypeptides with enhanced factor VIII activity are described. In some embodiments, the modified human factor VIII polypeptides comprise one or more amino acid substitutions at positions A20, T21, F57, L69, I80, L178, R199, H212, I215, R269, I310, L318, S332, R378, I610 and / or I661. Such polypeptides and viral vectors encoding such polypeptides may be used for treatment of FVIII deficiencies, such as hemophilia A.

An object of the present invention is to provide a novel cell for ex vivo gene therapy which produces a FVIII variant. The cell for ex vivo gene therapy of the present invention as a solution thereof produces a variant of human FVIII having the amino acid sequence at positions 20 to 2351 of the amino acid sequence of SEQ ID NO: 1 in which the B domain (the amino acid sequence at positions 760 to 1667) has been modified, thereby, the number of Asn residues subjected to N-linked glycosylation is four to six from the N-terminus and the function of the furin recognition site is lacking.

Provided are splice-modulating antisense oligonucleotides (ASOs) that target a terminal stem loop structure at the 3' end of intron 15 (TSL-3-15) of a Factor VIII (F8) pre-mRNA. Also provided are compositions comprising the splice-modulating ASOs. In some embodiments, the compositions are formulated for administration to a subject. Methods of treating Hemophilia A (HA) in a subject in need thereof are also provided. In certain embodiments, the subject exhibits aberrant splicing of Factor VIII (F8) exon 16, and the methods comprise administering to the subject a therapeutically effective amount of a composition of the present disclosure.

Provided are the polypeptide variants of the human factor VIII and polynucleotide sequences encoding the same optimized for gene therapy. Also provided are recombinant adeno-associated virus vectors for delivering the polypeptide variants and methods for treating type A hemophilia by using the said vectors.

The present disclosure provides, inter alia, methods of treating hemophilia A and computer-based systems, including software-based pharmacokinetic modeling systems, and their use for determining dosing information for subjects with hemophilia A.

A serum albumin variant or functional fragment thereof comprising one or more amino acid substitutions selected from the group consisting of: (i) glycine, isoleucine, lysine, methionine, phenylalanine, tryptophan, tyrosine, valine, and leucine substituting glutamine at position (522); (ii) valine substituting alanine at position (552); and (iii) alanine, glutamic acid, histidine, serine, lysine, and arginine substituting glycine at position (572).

To provide lentiviral vector (LV) formulations with improved stability and suitable for systemic administration, and pharmaceutical compositions comprising such LV formulations, and also to provide methods for treating disorders, especially blood disorders, using systemic administration of LV formulations.SOLUTION: Provided is a recombinant lentiviral vector preparation comprising: (a) a therapeutically effective amount of a recombinant lentiviral vector; (b) a TRIS-free buffer system; (c) a salt; (d) a surfactant; and (e) a carbohydrate, wherein the pharmaceutical composition is suitable for systemic administration to a human patient.SELECTED DRAWING: None