CA2634663C - Novel albumin-free factor viii formulations - Google Patents
Novel albumin-free factor viii formulations Download PDFInfo
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- CA2634663C CA2634663C CA002634663A CA2634663A CA2634663C CA 2634663 C CA2634663 C CA 2634663C CA 002634663 A CA002634663 A CA 002634663A CA 2634663 A CA2634663 A CA 2634663A CA 2634663 C CA2634663 C CA 2634663C
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/36—Blood coagulation or fibrinolysis factors
- A61K38/37—Factors VIII
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/02—Inorganic compounds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
- A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
- A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
- A61K47/183—Amino acids, e.g. glycine, EDTA or aspartame
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
- A61K47/38—Cellulose; Derivatives thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/04—Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S530/00—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof
- Y10S530/827—Proteins from mammals or birds
- Y10S530/829—Blood
Abstract
A Factor VIII composition formulated without albumin, comprising the following formulation excipients in addition to Factor VIII: 4% to 10% of a bulking agent selected from the group consisting of mannitol, glycine and alanine; 1% to 4% of a stabilizing agent selected from the group consisting of sucrose, trehalose, raffinose, and arginine; 1 mM to 5 mM calcium salt; 100 mM to 300 mM NaCl; and a buffering agent for maintaining a pH of approximately between 6 and 8. Alternatively, the formulation can comprise 2% to 6% hydroxyethyl starch; 1% to 4% of a stabilizing agent selected from the group consisting of sucrose, trehalose, raffinose, and arginine; 1 mM to 5 mM calcium salt; 100 mM to 300 mM NaCl; and a buffering agent for maintaining a pH of approximately between 6 and 8. In a further embodiment, the formulation can comprise: 300 mM to 500 mM NaCl; 1% to 4% of a stabilizing agent selected from the group consisting of sucrose, trehalose, raffinose, and arginine; 1 mM; 1 mM to 5 mM calcium salt; and a buffering agent.
Description
Novel Albumin-Free Factor VIII Formulations BACKGROUND OF THE INVENTION
Factor VM is a protein found in blood plasma which acts as a cofactor in the cascade of reactions leading to blood coagulation. A deficiency in the amount of Factor VIII activity in the blood results in the clotting disorder known as hemophilia A, an inherited condition primarily affecting males. Hemophilia A
is currently treated with therapeutic preparations of Factor VM derived from human plasma or manufactured using recombinant DNA technology. Such preparations are administered either in response to a bleeding episode (on-demand thernpy) or at fi-equent, regular intervals to prevent uncontrolled bleeding (prophylaxis).
Factor VIII is known to be zelatively unstable in therapeutic prepaiations. In blood plasma, Factor VIII is usually complexed with another plasma protein, von Willebrand factor (vWF), which is present in plasma in a large molar excess to Factor VIII and is believed to protect Factor VM from premahue degradation. Another circulating plasma protein, albumin, may also play a role in stabilizing Factor VM in vivo. Currently marketed Factor VIII preparations therefore primanly rely on the use of albumin and/or vWF to stabilize Factor VIII during the manufaGturing process and during storage.
The albumin and vWF used in currently marketed Factor VM preparations is derived from human blood plasma, however, and.the use of such material has certain drawbacks. Because a large molar excess of albumin compared to Factor VM is generally added in order to increase the stability of the Factor VM
in such preparations, it is difficult to characterize the Factor VIII protein itself in these preparations. The addition of human-derived albumin to Factor VM is WO 00/48635 PCT/US00/40068 , , .
Factor VM is a protein found in blood plasma which acts as a cofactor in the cascade of reactions leading to blood coagulation. A deficiency in the amount of Factor VIII activity in the blood results in the clotting disorder known as hemophilia A, an inherited condition primarily affecting males. Hemophilia A
is currently treated with therapeutic preparations of Factor VM derived from human plasma or manufactured using recombinant DNA technology. Such preparations are administered either in response to a bleeding episode (on-demand thernpy) or at fi-equent, regular intervals to prevent uncontrolled bleeding (prophylaxis).
Factor VIII is known to be zelatively unstable in therapeutic prepaiations. In blood plasma, Factor VIII is usually complexed with another plasma protein, von Willebrand factor (vWF), which is present in plasma in a large molar excess to Factor VIII and is believed to protect Factor VM from premahue degradation. Another circulating plasma protein, albumin, may also play a role in stabilizing Factor VM in vivo. Currently marketed Factor VIII preparations therefore primanly rely on the use of albumin and/or vWF to stabilize Factor VIII during the manufaGturing process and during storage.
The albumin and vWF used in currently marketed Factor VM preparations is derived from human blood plasma, however, and.the use of such material has certain drawbacks. Because a large molar excess of albumin compared to Factor VM is generally added in order to increase the stability of the Factor VM
in such preparations, it is difficult to characterize the Factor VIII protein itself in these preparations. The addition of human-derived albumin to Factor VM is WO 00/48635 PCT/US00/40068 , , .
2 also perceived as being a disadvantage with respect to recombinantly produced Factor VIII preparations. This is because recombinantly-derived Factor VIII
preparations, in the absence of such added albumin, would otherwise contain no human-derived proteins, and the theoretical risk of transmitting a virus would be 5 reduced.
Several attempts to formulate Factor VIII without albumin or vWF (or with relatively low levels of these excipients) have been described. For example, U.S. Patent No. 5,565,427 (EP 508 194) to Freudenberg (assigned to io Behnngwerke) describes Factor VIII prepatations which contain particular combinations of detergent and amino acids, specifically arginine and glycine, in addition to excipients such as sodium chloride and sucrose. The detergent, polysorbate 20 or polysorbate 80, is descnbed as being present in amounts of between 0.001 to 0.5% (v/v), while arginine and glycine are preseat in amounts 15 of between 0.01 to I mol/1. Sucrose is descnbed as being present in amounts of between 0.1 and 10%. Example 2 of this patent asserts that solutions of (1) 0.75% sucrose, 0.4 M glycine, and 0.15M NaC1, and (2) 0.01 M sodium citrate, 0.08 M glycine, 0.016M lysine, 0.0025 M calcium chloride, and 0.4 M sodium cliloride were not stable in solution over 16 hours, whereas solutions of (3) 1%, 2o sucrose, 0.14 M arginine, 0.1 M sodium chloride and (4) 1% sucrose, 0.4 M
glycine, 0.14 M arginine, 0.1 M sodium chloride, and 0.05% Tween 80 exhibited stability.
U.S. Patent No. 5,763,401 (EP 818 204) to Nayer (assigned to Bayer) also 25 descrn'bes a therapeutic Factor VIII formulation without albumin, comprising 15-60 mM sucrose, up to 50 mM NaCI, up to 5 mM calcium chloride, 65-400 mM glycine, and up to 50 mM histidine. The following specific formulations were identified as being stable: (1) 150 mM NaC1, 2.5 mM calcium chloride, and 165 mM mannitol; and (2) 1% sucrose, 30 mM sodium chloride, 2.5 mM
preparations, in the absence of such added albumin, would otherwise contain no human-derived proteins, and the theoretical risk of transmitting a virus would be 5 reduced.
Several attempts to formulate Factor VIII without albumin or vWF (or with relatively low levels of these excipients) have been described. For example, U.S. Patent No. 5,565,427 (EP 508 194) to Freudenberg (assigned to io Behnngwerke) describes Factor VIII prepatations which contain particular combinations of detergent and amino acids, specifically arginine and glycine, in addition to excipients such as sodium chloride and sucrose. The detergent, polysorbate 20 or polysorbate 80, is descnbed as being present in amounts of between 0.001 to 0.5% (v/v), while arginine and glycine are preseat in amounts 15 of between 0.01 to I mol/1. Sucrose is descnbed as being present in amounts of between 0.1 and 10%. Example 2 of this patent asserts that solutions of (1) 0.75% sucrose, 0.4 M glycine, and 0.15M NaC1, and (2) 0.01 M sodium citrate, 0.08 M glycine, 0.016M lysine, 0.0025 M calcium chloride, and 0.4 M sodium cliloride were not stable in solution over 16 hours, whereas solutions of (3) 1%, 2o sucrose, 0.14 M arginine, 0.1 M sodium chloride and (4) 1% sucrose, 0.4 M
glycine, 0.14 M arginine, 0.1 M sodium chloride, and 0.05% Tween 80 exhibited stability.
U.S. Patent No. 5,763,401 (EP 818 204) to Nayer (assigned to Bayer) also 25 descrn'bes a therapeutic Factor VIII formulation without albumin, comprising 15-60 mM sucrose, up to 50 mM NaCI, up to 5 mM calcium chloride, 65-400 mM glycine, and up to 50 mM histidine. The following specific formulations were identified as being stable: (1) 150 mM NaC1, 2.5 mM calcium chloride, and 165 mM mannitol; and (2) 1% sucrose, 30 mM sodium chloride, 2.5 mM
3 calcium chloride, 20 mM histidine, and 290 mM glycine. A formulation containing higher amounts of sugar (10% maltose, 50 mM NaCI, 2.5 mM
calcium chloride, and 5 mM histidine) was found to exhibit poor stability in the lyophilized state compared with formulation (2).
U.S. Patent No. 5,733,873.(EP 627 924) to Osterberg (assigned to Pharmacia &
Upjohn) discloses formulations which include between 0.01 - I mg/ml of a surfactant. This patent discloses formulations having the following ranges of excipients: polysorbate 20 or 80 in an amount of at least 0.01 mgJml, preferably 0.02 - 1.0 mg/ml; at least 0.1 M NaCI; at least 0.5mM calcium salt; and at least 1 mM histidine. More particularly, the following specific formulations are disclosed: (1) 14.7 - 50 - 65 mM histidine, 0.31- 0.6 M NaCI, 4 rnM calcium chloride, 0.001 - 0:02 - 0.025% polysorbate 80, with or without 0.1% PEG 4000 or 19.9 mM sucrose; and (2) 20 mg/ml mannitol, 2.67 mg/ml histidine, 18 11 mg/ml NaCI, 3.7 mM calcium chloride, and 0.23 mg/ml polysorbate 80.
., . .
Other attempts to use low or high concentrations of sodium chloride have also been described. U.S. Patent No. 4,877,608 (EP 315 968) to Lee (assigned to '. ' Rhone-Poulenc Rorer) teaches formulations with relatively low concentrations of sodium chloride, namely formulations comprising 0.5 mM - 15 mM NaCI, 5 mM calcium chloride, 0.2 mM - 5 mM histidine; 0.01 - 10 mM lysine hydrochloride and up to 10% sugar. The "sugar" can be up to 10% maltose, 10=o sucrose, or 5% mannitol.
US 5,605,884 (EP 0 314 095) to Lee (assigned to Rhone-Poulenc Rorer) teaches the use of formulations with relatively high concentrations of sodium chloride.
These formulations include 0.35 M -1.2 M NaCl,1.5 - 40 mM calcium chloride, 1 mM - 50 mM histidine, and up to 10% of a"sugar such as mannitol, sucrose,
calcium chloride, and 5 mM histidine) was found to exhibit poor stability in the lyophilized state compared with formulation (2).
U.S. Patent No. 5,733,873.(EP 627 924) to Osterberg (assigned to Pharmacia &
Upjohn) discloses formulations which include between 0.01 - I mg/ml of a surfactant. This patent discloses formulations having the following ranges of excipients: polysorbate 20 or 80 in an amount of at least 0.01 mgJml, preferably 0.02 - 1.0 mg/ml; at least 0.1 M NaCI; at least 0.5mM calcium salt; and at least 1 mM histidine. More particularly, the following specific formulations are disclosed: (1) 14.7 - 50 - 65 mM histidine, 0.31- 0.6 M NaCI, 4 rnM calcium chloride, 0.001 - 0:02 - 0.025% polysorbate 80, with or without 0.1% PEG 4000 or 19.9 mM sucrose; and (2) 20 mg/ml mannitol, 2.67 mg/ml histidine, 18 11 mg/ml NaCI, 3.7 mM calcium chloride, and 0.23 mg/ml polysorbate 80.
., . .
Other attempts to use low or high concentrations of sodium chloride have also been described. U.S. Patent No. 4,877,608 (EP 315 968) to Lee (assigned to '. ' Rhone-Poulenc Rorer) teaches formulations with relatively low concentrations of sodium chloride, namely formulations comprising 0.5 mM - 15 mM NaCI, 5 mM calcium chloride, 0.2 mM - 5 mM histidine; 0.01 - 10 mM lysine hydrochloride and up to 10% sugar. The "sugar" can be up to 10% maltose, 10=o sucrose, or 5% mannitol.
US 5,605,884 (EP 0 314 095) to Lee (assigned to Rhone-Poulenc Rorer) teaches the use of formulations with relatively high concentrations of sodium chloride.
These formulations include 0.35 M -1.2 M NaCl,1.5 - 40 mM calcium chloride, 1 mM - 50 mM histidine, and up to 10% of a"sugar such as mannitol, sucrose,
4 or maltose. A fonnulation comprising 0.45 M NaCI, 2.3 mM calcium chloride, and 1.4 mM histidine is exemplified.
International Patent Application WO 96/22107 to Roser (assigned to Quadrant Holdings Cambridge Limited) describes formulations which include the sugar trehalose. These fonnulations comprise: (1) 0.1 M NaC1, 15 mM'calcium chloride, 15 mM histidine, and 1.27 M (48%) trehalose; or (2) 0.011 % calcium chloride, 0.12% histidine, 0.002% Tris, 0.002% Tween 80, 0.004% PEG 3350, 7.5% trebalose, and either 0.13% or 1.03% NaCI.
Other therapeutic Factor VIII formulations of the prior art generally include albumin and/or vWF for the purpose of stabilizing Factor VIII and are therefore not relevant to the present invention. For example, U.S. Patent No. 5,328,694 (EP 511234) to Schwinn (assigned to Octapharma AG) descn'bes a fonnulation which includes 100 - 650 mM disaccharide and 100 mM- 1.0 M amino acid.
Specifically, the following formulations are disclosed: (1) 0.9 M sucrose, 0.25 =, .
M glycine, 0.25 M lysine, and 3 mM calcium chloride; and (2) 0.7 M sucrose, 0.5 M glycine, and 5 mM calcium chloride.
While several attempts have been made to formulate Factor VIII without albumin or vWF, there remains a nced for therapeutic Factor VIII formulations which are stable in the absence of albumin or other proteins.
SUMMARY OF THE INVENTION
The present invention relates to therapeutic Factor VIII compositions which are stable in the absence of albumin. In particular, the present invention comprises a Factor VIII composition comprising, in addition to Factor VIII: 4% to 10 %
of a bulking agent selected from the group consisting of mannitol, glycine and alanine; 1% to 4% of a stabilizing agent selected from the group consisting of
International Patent Application WO 96/22107 to Roser (assigned to Quadrant Holdings Cambridge Limited) describes formulations which include the sugar trehalose. These fonnulations comprise: (1) 0.1 M NaC1, 15 mM'calcium chloride, 15 mM histidine, and 1.27 M (48%) trehalose; or (2) 0.011 % calcium chloride, 0.12% histidine, 0.002% Tris, 0.002% Tween 80, 0.004% PEG 3350, 7.5% trebalose, and either 0.13% or 1.03% NaCI.
Other therapeutic Factor VIII formulations of the prior art generally include albumin and/or vWF for the purpose of stabilizing Factor VIII and are therefore not relevant to the present invention. For example, U.S. Patent No. 5,328,694 (EP 511234) to Schwinn (assigned to Octapharma AG) descn'bes a fonnulation which includes 100 - 650 mM disaccharide and 100 mM- 1.0 M amino acid.
Specifically, the following formulations are disclosed: (1) 0.9 M sucrose, 0.25 =, .
M glycine, 0.25 M lysine, and 3 mM calcium chloride; and (2) 0.7 M sucrose, 0.5 M glycine, and 5 mM calcium chloride.
While several attempts have been made to formulate Factor VIII without albumin or vWF, there remains a nced for therapeutic Factor VIII formulations which are stable in the absence of albumin or other proteins.
SUMMARY OF THE INVENTION
The present invention relates to therapeutic Factor VIII compositions which are stable in the absence of albumin. In particular, the present invention comprises a Factor VIII composition comprising, in addition to Factor VIII: 4% to 10 %
of a bulking agent selected from the group consisting of mannitol, glycine and alanine; 1% to 4% of a stabilizing agent selected from the group consisting of
5 PCT%US00/40068 sucrose, trehalose, raffinose, arginine; 1 mM to 5 mM calcium salt; 100 inM to .
300 mM NaCI; and a buffering agent for maintaining a pH of approximately between 6 and 8. This composition can additionally comprise a surfactant such as polysorbate 20, polysorbate 80, Pluronic F68, or Brij 35. When the 5 surfactant is polysorbate 80, it should be present in an amount of less than 0_1%. -The buffer of the Factor VIII compositions according to the present invention is preferably present in a concentration of from 10 mM to 50 mM, and is preferably selected from the group consisting of histidine, Tris, BIS-Tris Propane, PIPES, MOPS, HEPES, MES and ACES. Advantageously, the buffering agent is either histidine or Tris. The Factor VIII composition of the present invention can further comprise an antioxidant.
The Factor VIII compositions of the present invention include both a bulking agent and a stabilizer. The bulking agent can be present in an amount of from about 6% to about 8=/., preferably about 8%. TLe stabilizing agent is preferably present in an amount of about 2%. Sodium cliloride is also present in these compositions, preferably in an amount of from 150 to 350 mM, and more preferabiy in an amount of about 225 mM. The calcium salt of the composition is also preferably calcium chloride, and the composition itself is preferably in lyophilized form.
In another embodiment, the present invention can comprise a Factor VIII
composition formulated without adding albumin which includes the following excipients in addition to Factor VIII: 2% to 6 % hydroxyethyl starch; 1% to 4%
of a stabilizing agent selected from the group consisting of sucrose, trehalose, raffinose, arginine; l mM to 5 mM calcium salt; 100 mM to 300 mM NaCI;
and a buffering agent for maintaining a pH of approximately between 6 and 8.
WO 00/48635 pCT/US00/40068
300 mM NaCI; and a buffering agent for maintaining a pH of approximately between 6 and 8. This composition can additionally comprise a surfactant such as polysorbate 20, polysorbate 80, Pluronic F68, or Brij 35. When the 5 surfactant is polysorbate 80, it should be present in an amount of less than 0_1%. -The buffer of the Factor VIII compositions according to the present invention is preferably present in a concentration of from 10 mM to 50 mM, and is preferably selected from the group consisting of histidine, Tris, BIS-Tris Propane, PIPES, MOPS, HEPES, MES and ACES. Advantageously, the buffering agent is either histidine or Tris. The Factor VIII composition of the present invention can further comprise an antioxidant.
The Factor VIII compositions of the present invention include both a bulking agent and a stabilizer. The bulking agent can be present in an amount of from about 6% to about 8=/., preferably about 8%. TLe stabilizing agent is preferably present in an amount of about 2%. Sodium cliloride is also present in these compositions, preferably in an amount of from 150 to 350 mM, and more preferabiy in an amount of about 225 mM. The calcium salt of the composition is also preferably calcium chloride, and the composition itself is preferably in lyophilized form.
In another embodiment, the present invention can comprise a Factor VIII
composition formulated without adding albumin which includes the following excipients in addition to Factor VIII: 2% to 6 % hydroxyethyl starch; 1% to 4%
of a stabilizing agent selected from the group consisting of sucrose, trehalose, raffinose, arginine; l mM to 5 mM calcium salt; 100 mM to 300 mM NaCI;
and a buffering agent for maintaining a pH of approximately between 6 and 8.
WO 00/48635 pCT/US00/40068
6 Preferably, such a composition comprises about 4% hydroxyethyl starch, and the NaCI is present in an amount of 200 mM. The stabilizing agent is also preferably present in an amount of about 2%.
In a further embodiment, the present invention includes a Factor VIII
composition, formulated without albumin, comprising: 300 mM to 500 mM
NaC1;1 % to 4% of a stabilizing agent selected from the group consisting of sucrose, trehalose, raffinose, arginine; 1 mM to 5 mM calcium salt; and a buffering agent for maintaining a pH of approximately between 6 and 8.
1 o Preferably, the NaCI is present in a concentration of about 400 mM.
In yet another embodiment, the present invention comprises a process for lyophilizing an aqueous Factor VIII composition in a container using a lyophilizer, wherein the process comprises an initial freezing step, and the initial freezing step further comprises the steps of (a) lowering the tempcrature of the lyophilizer chamber to at least about -45 C; (b) raising the t,emperature of the chamber to between about -15 C and -25 C; and subsequently (c) lowering the temperattire of the chamber to at least about -45 C. In this process, the temperature of the chamber is preferably lowered or raised at a rate of between 2o about 0.5 Cand about 1.0 C per minute. In step (a), the temperature is preferably maintained for about 1 hour, and is lowered to about -55 C. In step (b) the temperature is preferably maintained be -15 C and -25 C for between I
and 3 hours, and more preferably is at -22 C, and the temperature in step (c) is -preferably maintained for about 1 hour. The Factor VIII composition used in this process preferably comprises between 4% and 10 % of an agent selected from the group consisting of mannitol, glycine and alanine, and also preferably comprises between 1% and 4% of an agent selected from the group consisting of sucrose, trehalose, raffuiose, and arginine. In addition, the Factor VIII
In a further embodiment, the present invention includes a Factor VIII
composition, formulated without albumin, comprising: 300 mM to 500 mM
NaC1;1 % to 4% of a stabilizing agent selected from the group consisting of sucrose, trehalose, raffinose, arginine; 1 mM to 5 mM calcium salt; and a buffering agent for maintaining a pH of approximately between 6 and 8.
1 o Preferably, the NaCI is present in a concentration of about 400 mM.
In yet another embodiment, the present invention comprises a process for lyophilizing an aqueous Factor VIII composition in a container using a lyophilizer, wherein the process comprises an initial freezing step, and the initial freezing step further comprises the steps of (a) lowering the tempcrature of the lyophilizer chamber to at least about -45 C; (b) raising the t,emperature of the chamber to between about -15 C and -25 C; and subsequently (c) lowering the temperattire of the chamber to at least about -45 C. In this process, the temperature of the chamber is preferably lowered or raised at a rate of between 2o about 0.5 Cand about 1.0 C per minute. In step (a), the temperature is preferably maintained for about 1 hour, and is lowered to about -55 C. In step (b) the temperature is preferably maintained be -15 C and -25 C for between I
and 3 hours, and more preferably is at -22 C, and the temperature in step (c) is -preferably maintained for about 1 hour. The Factor VIII composition used in this process preferably comprises between 4% and 10 % of an agent selected from the group consisting of mannitol, glycine and alanine, and also preferably comprises between 1% and 4% of an agent selected from the group consisting of sucrose, trehalose, raffuiose, and arginine. In addition, the Factor VIII
7 composition used in this process also preferably comprises between 100 mM and 300 mM NaCI.
In accordance with an aspect of the present invention, there is provided a Factor VIII composition formulated without adding albumin to said composition, comprising the following formulation excipients in addition to Factor VIII: 2%
to 6% hydroxyethyl starch; 1% to 4% of a stabilizing agent selected from the group consisting of sucrose, trehalose, raffinose, and arginine; 1 mM to 5mM calcium salt; 100 mM to 300 mM NaCI; and a buffering agent for maintaining a pH of approximately between 6 and 8.
DETAILED DESCRIPTION OF THE INVENTION
Definitions As used herein, the terms below and variations thereof shall be defined as follows, unless otherwise indicated:
Factor VIII - The Factor VIII molecule exists naturally and in therapeutic preparations as a heterogeneous distribution of polypeptides arising from a single gene product (see, e.g., Anderson et al., Proc. Natl. Acad. Sci. USA, 83, 2979-2983, May 1986). The term "Factor VIII" as used herein refers to all such polypeptides, whether derived from blood plasma or produced through the use of recombinant DNA techniques. Commercially available examples of therapeutic preparations containing Factor VIII include those sold under the trade names of HEMOFIL M and RECOMBINATE (available from Baxter Healthcare Corporation, Deerfield, Illinois, U.S.A.). Other preparations currently in development comprise primarily a single subpopulation of Factor VIII molecules which lack the B domain portion of the molecule.
7a International Unit, IU- International Unit, or IU, is a unit of measurement of the blood coagulation activity (potency) of Factor VIII as measured by a standard assay, such as one of the following:
One stage assay. One stage assays are known to the art, such as that described in Lee, Martin L, et al., An Effect of Predilution on Potency Assays of Factor VIII Concentrates, Thrombosis Research (Pergamon Press Ltd.) 30, 511-519 (1983).
S
Chromogenic assay. Chromogenic assays maybe purchased commercially, sueh as the Coatest Factor VIII, available from Chromogenix AB, Molndal, Sweden.
Anneal The term anneal shall be used to indicate a step in the lyophilization process of a pharmaceutical preparation undergoing lyophilization, prior io the freeze-drying of the preparation, in which the temperature of the preptation is . ~\
raised from a]ower temperature to a higher temperature and then cooled again after a period of time.
Bulking Agent - For the purposes of this application, bulking agents are lhose chemical entities which provide structure to the "cake" or residual solid mass of a pharmaceutical preparation after it has been lyophilized and which protect it ag.ainst collapse. A crystallizable bulking agent shall mean a bulking agent as descn'bed herein which can be crystallizcd during lyophilization, other than sodium chloride. HES isnot included in this group of crystallizable bullang agents.
Freeze-drying, f freezin& lyophilizing -. 'Troeez.e-drying," unless otherwise indicated by the conteat in which it appens, shall be used to denote the portion of a lyophilization process in which the temperature of a pliarmaceutical preparation is raised in order to drive water out of the preparatioyn'. Th,e "freezing" steps of a lyophilization pmoess.are those steps which occur prior to the freez.e-drying stage. "Lyophilizing," unless othrawise indicated, shall refer 2S to the entire process of lyophilization, including both the Erees.ing steps and the freeze-drying steps.
Unless otherwise noted, percentage terms express weight/volume percentages and temperatures are in the Celsius scale.
Formulation Components The Factor VIII compositions of the present invention include bulking agents, stabilizing agents, buffering agents, sodium chloride, calcium salts, and, advantageously, other excipients. These excipients have been chosen in order to maximize the stability of Factor VIII in lyophilized preparations.. However, the Factor VIII compositions of the present invention exhibit stability in the liquid state as well.
The bulking agents used in the pn.sent formulations, which form the crystalline portion of the lyophilized product (except in the case of HES), are selected from the group consisting of mannitol, glycine, alanine, and hydroxyethyl starch (IM). Mannitol, glycine, or alanine are present in an amount of 4 - 10%, preferably 6- 9'/0, and more preferably about 8'/o. When HES is used as a bulking agent, it is present in an amount of 2 - 6%, preferably 3- 5%, and more preferably about 4%.
The stabilizing agents used in the formulations of the present invention are selected from the group consisting of sucrose, trehalose, raffinose, and arginine.
2o These agents are present in the formulations of the present invention in an atnount of between 1- 4%, preferably 2 - 3%, more preferably about 2%.
Sorbitol and glycerol were evaluated as possible stabilizers but were found to be poor stabilizers in the present formulations.
7,5 . Sodium chloride is included in the present formulations in an amount of 300 mM, preferably 150 - 250 mNrl, and most preferably about 225 mM. In one embodiment of the present invention, sodium chloride itself can be used without any of the aforementioned bulking agents, in which case it would be included in the fonnuiation in an amount of between 300 mM and 500 mM NaCI, preferably 350 to 450 mM NaCI, and more preferably about 400 mM NaCI.
In addition, buffers are present in these fonnulations, because it is believed that 5 the Factor VIII molecule can be adversely affected by pH shifts during lyophilization. The pH should preferably be maintained in the range of between 6 and 8 during lyophilization, and more preferably at a pH of about 7. The buffering agent can be any physiologically acceptable chemical entity or combination of chemical entities which have the capacity to act as buffers, 1 o including histidine, Tris, BIS-Tris Propane, PIPES, MOPS, HEPES, MES and ACES. The full cbeinical designations of these buffering agents is listed in Table I below. Typically; the buffering agent is included in a concentration of 10 - 50 mM. When histidine is added to the formulations, concentrations of over 20 mM and preferably about 25 mM are used, alone or in combination vvith other buffers such as Tris. Histidine is especially preferred for use in the compositions of the present invention, as descn'bed in greater detail below.
, ..= ' .
Table 1- Buffering Agents Tris tris-(hydroxymethyl)-aminomethane BIS-Tris Propane 1,3-bis-(tris-(hydroxy-methyl)methylamino) propane PIPES pipera2ine-N,N'-bis-(2-ethanesulfonic acid) MOPS 3-(N-morpholino) propanesulfonic acid HEPES N-2-hydroxyethyl-piperazine-N'-2-ethanesulfonic acid MES 2-(N-morpholino) ethanesulfonic acid ACES N-2-acetamido-2-aminoethanesulfonic acid In. order to preserve the activity of Factor VIII, it is inzportant that the formulations of the present invention also include calcium or another divalent cation able to interact with Factor VIII and maintain its activity, presumably by maintaining the association of the heavy and light chains of Factor VIII.
Between 1 mM and 5 mM of a calcium salt can be used, more preferably 3- 4 mM, and most preferably about 4 mM. The calcium salt is preferably calcium chloride, but can also be other calcium salts such as calcium gluconate, calcium glubionate, or calcium gluceptate.
The Factor VIII compositions of the present invention also preferably include a surfactant, preferably in an amount of 0.1 Jo or less, and more preferably in an amount of about 0.03%. The surfactant can, for example, be chosen from the group consisting of polysorbate 20, polysorbate 80, pluronic polyols, and Brij 35 (polyoxyethylene 231auryl ether). Several grades of pluronic polyols (sold under the trade name Pluronic, manufactured by the BASF Wyandotte Corporation) are available. These polyols, of diversified molecular weight (from 1,000 to over 16,000) and physicochemical properties have been used as surfactants. Pluronic F-38, of a molecular weight of 5,000 and Pluronic F-68, molecular weight 9,000, both contain (by weight) 80 per cent hydrophilic polyoxyethylene groups and 20 percent hydrophobic polyoxypropylene groups.
> = .
Tween-80, a commercial polysorbate, however, is preferred in the present formulations, in particular vegetable-derived Tween-80.
The Factor VIII formulations of thepresent invention also preferably include an antioxidant. The addition of antioxidants to the lyophilized formulations of the invention has been found to improve the stability of these formulations, and thus extend their shelf lives. The antioxidants used must be compatible for use with a pharmaceutical preparation, and in addition are preferably water soluble.
When adding antioxidants to a formulation, it is preferable to add such antioxidants as late in the process prior to lyophilization as possible, in order to avoid spontaneous oxidation of the antioxidant. Table 2 below lists suitable antioxidants, which are available commercially through companies such as Calbiochem and Sigma.
Table 2 - Antioxidants N-Acetyl-L-Cysteine / Homocysteine Glutathione -6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) Lipoic acid Methionine Sodium Thiosulfate Platinum Glycine-glycine-histidine (tripeptide) Butylatedbydroxytoluene (BITT) Of the foregoing antioxidants, glutathione is prefen=ed. Concentrations in the range of about 0.05 mg/mI to more than 1.0 mgtml have all been found to ',. enhance the stability of Factor VIII compositions, and it is believed that higher concentrations would also be useful (up to the point of any toxic effects or adverse manufacturing effects, such as a depression of the glass transition temperature of the lyophilized product).
It has been found in particular that the combination of histidine and glutathione produces synergistically beneficial effects on the stability of Factor VIII
compositions. Histidine, while acting as a buffer, can also act as a metal chelator. To the extent that Factor VIII inactivation is caused by metal-induced oxidation, histidine can therefore act to stabilize Factor VIII by binding such oxidizing metal ions. It is believed that by binding these metals, the glutathione (or indeed any other antioxidant present) is thereby able to provide fnrther antioxidative protection, since the oxidative effect of the metal ions bound by.
the histidine has been contained.
Other chelating agents might also be used in the compositions of the present invention. Such agents should preferably bind metals such as copper and iron with greater affinity than calcium, if a calcium salt is being used in the composition. One such chelator is deferoxamine, a chelating agent that facilitates the removal of AI++ and iron. Deferoxamine Mesylate, C25H48N608*CH4O3S, can be obtained from Sigma (Sigma Prod. No.
D9533). It is an aluminum and iron(II) chelator which chelates iron (as a 1:1 chelate complex) only in the +3 oxidation state, not +2 oxidation state, and ean also bind manganese ion and other metals. Deferoxamine can be used advantageously in an amount of 0.25 mg/1.
The Factor VIII used in the present formulations can be either highly purified human plasma-derived Factor VIII or more preferably can be recombinantly =~
produced Factor VIII. Recombinant Factor vIII can be produced by Chinese hamster ovary (CHO) cells transfected with a vector canying a DNA sequence coding for the Factor VIII molecule. Methods for creating such transfected CHO
cells are described, inter alia, in U.S. Patent No. 4,757,006 to Toole, Jr., though alternative methods are also known to the art (see, e.g., U.S. Patent No.
4,868,112, also to Toole, Jr., and PCT International Application WO A-91/09122). The methods used to culture sucb CHO cells to produce Factor VIII
are also lcnown to the art, for example in European Patent Application No. 0 218 to Genetics Institute, entitled "Improved method for producing Factor VIII:C-type proteins." Recombinant Factor VIII can, however, also be produced in other cell lines, such as baby hamster kidney (BHK) cells. The Factor VIII
molecule itself, if recombinantly produced, can be either full-length Factor VIII
or a deletion derivative thereof, such as a B domain-deleted Factor VIII
molecule.
While the Factor VIII compositions descnbed in this application can be lyophilized and reconstituted in the indicated concentrations, one of skill in the art will understand that these preparations can also be reconstituted in more dilute form. For example, a preparation according the present invention which is lyophilized and/or normally reconstituted in 2 ml of solution can also be reconstituted in a larger volume of diluent, such as 5 ml. This is particularly appropriate when the Factor VIII preparation is being injected into a patient immediately, since in this case the Factor VIII is less likely to lose activity, -which may occur more rapidly in more dilute solutions of Factor VIII.
Formulation and LyopbiGzation Development In order to achieve maximal stability, the Factor VIII compositions of the present invention are preferably lyophilized. During lyophilization, Factor VIII
is converted from being in an aqueous phase to being in an asnorphous solid t, phase, which is thought to protect the protein from chemical and/or confonnational instability. The lyophilized preparation not only contains an amorphous phase, but also includes a coiinponent wluch crystallizes during lyophilization. This is thought to allow the rapid lyophilization of the Factor VIII composition and the fonnation.of a more elegant cake (that is, a cake with minimal shrinkage from the sides of the container in which it was lyophilized).
In the formulations of the present invention, the stabilizing agents have been selected to exist primarily in an amorphous phase of the lyophilized product, while the bulking agents (except HES) have been selected to crystallize during freezing.
WO 00/48635 PCT/[1S00/40068 Both the Factor VIII and the stabilizer are preferably dispersed in the amorphous phase of the lyophilized cake. The mass of the stabilizer is also preferably large compared to the other excipients in the amorphous form. In addition, the apparent glass transition temperature (Ts') of the amorphous phase 5 is preferably relatively high during freeze-drying, and the glass transition temperature (Tg) of the solid is likewise preferably high during storage.
Crystallization of sodium chloride in the product was found to be desirable, since amorphous sodium cbloride will depm,ss the T.' of the amorphous phase.
10 In order to avoid the collapse of the cake of a particular composition, primary drying is preferably carried out at a product temperature below the apparent glass transition temperature of the freeze concentrate. . An increase in drying time may also be required to offset a decrease in Ts'. Further infonnation on lyophilization may be found in Carpenter, J.F. and Chang, B.S., Lyophi7izatinn 15 ofProtein Pharmaceuticals, BiotechnoloQV and Biopharmaccutical Manufacturing, Processing and Preservation, K.E. Avis and V.L. Wu, eds.
(Buffalo Grove, IL: Interpharm Press, Inc.), pp. 199-264 (1996).
;
Example 1 The effects of the concentration of Factor VIII and of the addition of a stabilizer on the recovery of Factor VIII were investigated in several studies. These studies were perfonmed using mannitol as a model bulking agent and sucrose as a model stabilizer. The three sample formulations described in Table 3 below were used in these studies. All formulations used in these studies included 10 mM Tris, 200 mM NaCI, 8% mannitol, 4 mM CaCl2, and 0.02% Tween-80 and were conducted at pH 7Ø
Table 3 Sample Initial Factor VIII Sucrose %
I.D. (ILT/m!) IA. 600 1iese samples were lyophilized using the freeze-drying cycle shown in Table 4 below in order to maintain a product temperature below the apparent glass s transition temperature (T,'). Differential scanning calorimetric (DSC) studies indicated the presence of a transition at approximately -40 C in the mannitol formulations. In order to maintain a product temperature below this value, the shelf temperature was set to -32 C during primary drying. Primary drying under these conditions was perfon:ned for about 55 hours, with a total cycle time io of about 80 hours.
,., Table 4 Freezing/Processing Description Metbod I Cool to +5 C;
(Freezing) Cool to -5 C at 1 C/minute, hold for 20 minutes;
Cool to -20 + 5 C at 1 C/minute; hold for 1 hour (up to 3 hours);
Cool to -45 C at 0.5 C/minute, hold for 1 hour.
II Freeze per method I
Hold at -35 C for 48 hours.
m Freeze per method I
Hold at -35 C for 48 hours;
Hold at 20 C for 48 hours.
IV Shelf -32 C during primary drying for (Freeze-drying) about 55 hours (up to 100 hours);
Product < -40 C during primary drying;
Ramp from -32 C to +40 C at 0.2 C/minute;
Shelf +40 C during secondary drying for 3 hours.
-, , The Factor VIII activity of these samples, as determined by the one-stage clotting assay, was compared against a control held at -45 C. The assay results are shown in Table 5 below.
Table 5 Processing % Loss in Factor VIII Activity During Each Step Method Formulation IA Formulation IB Formulation IC
(600 IU/ml) (60 IU/ml) (60 IU/ml, 2%
Sucrose) 1 6.7 37.5 41.7 II. 2.0 9.3 3.9 III 7.3 11.6 5.0 N 20.0 24.2 18.3 (Lyophilization) These results indicate that protein concentration has an effect on the recovery of Factor VIII during freezing. Formulations containing 60 IU/ml lost approximately 37 lr-42 !0 of the initial Factor VIII activity during the freezing step, while 6.7% of Factor VIII activity was lost for the formulation containing 600 IU/ml. These results indicate that a higher protein concentration has a protective effect during ficezzing. Although sucrose provided some protection to the Factor VIII during the intermediate tonpera'ture holds as well as during freeze-drying, it failed to protect the protein during the initial freezing step.
Example 2 Following the development of the lyophilization process outlined in Example 1, further optimization of this process was undertaken. It has been found that a lyophilized composition having a higher glass transition temperature, (and, theoretically, better Factor VIII stability) can be produced by: (1) lowering the freezing temperature initially to -45 C or lower (such as down to about -50 C
or -55 C); (2) raising the temperature to -20 C or -22 C (+5 C); and then (3) lowering the temperature again to -45 C or lower. The temperature is lowered or raised, as the case may be, at a rate of between about 0.5 C and about 1.0 C
per minute. Once the desired temperature is reached, the composition is held at that temperature for between I and 3 hours. This improved freezing cycle is shown in Table 6 below.
Table 6 Freezing Method Description I Cool to +5 C;
Cool to -5 C at 0.5-1 Clminut.e, hold for 20 niinutes;
Cool to between-55 C and -45 C at 0.5-1 C/minute, hold for about 1 hour, Warm to -22 C ( 5 C) at 0.5-1 CJminute, bold for I to 3 hours;
Cool tio -45 C at 0.5-1 C/minute, hold for about 1 hour.
Unless otherwise indicated, the temperatures referred to in this example and in other examples refer to the shelf temperature of the lyophilizer and not to the temperature of the product per se. Following the improved freezing cycle, the remainder of the lyophilization process can be conducted as outlined in Example 1 above, or otherwise as described fiuther herein or as determined by one of skill in the art.
This improved lyophilization process was found to be useful for fonnulations which include glycine as the bulking agent as well as those which use mannitol.
It is fnrther believed to have applicability to formulations which maice use of the other bullcing agents of the present invention as well.
WO 00/48635 PCT/US00l40068 Example 3 It is believed that in order to produce a freeze-dried product with acceptable cake appearance and glass transition temperature, the bulking agent of lyophilized pharmaceutical preparations which contain sodium chloride, such as 5 glycine or mannitol, may need to be crystallized. The following improved lyophilization process for crystallizable bulking agents was therefore developed.
Table 7a - Freezing Steps Process Step Temperature Duration of Step Initial freezing -40 C or less 1 hour First annealing between -23 C and 27 C 3 hours Second freeyin,g -55 C 1 hour Second annealing -36 C 4 bours Third freezing -50 C 1 hour ' ~ . Table 7b - Freeze-Drying Steps Process Step Temperature Duration of Step Primary Drying -35 C up to 100 hours Secondary Drying: First step 40 C 3 hours }
Secondary Drying: Second step 45 C 3 hours Secondary Drying: Third step 50 C 3 hours In the freezing steps, changes in the temperatures occurred at a rate of between about 0.5 C/minute and 1 C/minute.' It is believed that steps of longer duration would also be effective.
Prior to the first freezing step, the temperature is brought to between about and 8 C for about one hour for the purpose of bringing all the vials to approximately the same temperature. After this the lyophilizer is cooled to -C. The first freezing step should be perfonned at a temperature less than -5 30 C, preferably below -35 C, and more preferably at about -40 C: Following this, the first annealing step should occur at a temperature of between -30 C
and -19 C, more preferably either between about 25 C and -28 C (if glycine is the bulking agent) or between -21 C and 24 C (if mannitol is the bulking agetit), with the temperatures of 23 C and 26 C being most preferred, at which temperatures it is believed that the crystallizable bulking agents crystallize, at least in part. However, the lower range around -27 C is not recommended for formulations containing mannitol and arginine. This step is preferably carried out for about 3 hours.
Following the first annealing step, the temperature is lowered, preferably to less than about -50 C and more preferably to less than -55"C, for about 1 hour. It is believed that the sodium chloride in the preparation nucleates at this time.
During the second annealing step, the temperature of the pharmaceutical preparation is raised to between about --30 C and -39 C, and preferably to about -33 C for rnannitol-containing compositions and -36 C for glycine-containing compositions. It is believed that NaCI crystal growth occurs at this time, at least in part. This step is preferably conducted for about 4 hours. Following this, the temperature of the lyophilizer is reduced to about -50 C, preferably for about hour in order to reduce the temperature of the preparation.
In the freeze-drying steps which follow, changes in temperature occurred at a rate of between about 0.1 C/minute and 0.5 C/minute. After reducing the pressure in the lyophilizer to about'65 mTorr, the temperature is raised to between about -32 C and -35 C for primary drying. Ice crystals in the preparation will sublimate at this temperature. This step is performed for up to about 100 hours, or until most of the ice has been sublimated from the preparation. The point at which most of the ice has sublimated can be determined, for example, using a dewpoint sensor, which indicates the end of the sublimation of ice when the readings decrease (the point of inflection).
Following primary drying, the temperature is raised to +40 C, preferably at a rate of 0.2 Clminute, to initiate secondary drying to remove fiuther water from the preparation. This temperature is preferably maintained for about three hours. Second and third secondary drying steps follow this first step, where the temperature is raised to about +45 C for about three hours and then to about +50 C for three more hours in order to reduce the moisture in the lyophilized cake to less than 2% (w/w).
Eiample 4 Further studies were performed to examine specifically the effect of histidine on lyophilized Factor VIII compositions containing glycine or mannitol as bulking agents. Non-reversing beat flow (Modulated DSC, mDSC) was used to detect the crystallization of these bulking agents during cooling. Both the temperature of crystallization and the total heat of crystallization were determined from the crystallization exotherm. The appearance of the NaCI eutectic melt endotherm during warming was used to detect NaCI crystalliza6on. In mDSC, the extent of crystallization was determined as the ratio of the enthalpy of melting of the formulation to the enthalpy of melting of pure NaCI solution by using the total heat flow signal. In addition, X-ray diffraction analyses were performed in order to determine the extent of crystallization in the lyophilized formulations.
WO 00/48635' PCT/US00/40068 While histidine concentrations less than 20 mM did not significantly impact the crystallization of glycine, 50 mM histidine reduced the extent of glycine crystallization. Well-defined NaCI crystallization exotherms were not observed during cooling of formulations containing glycine. However, eutectic melting endotherms during heating indicated that NaCI was crystallized (> 50%) after cooling lower than 50 C and annealing at 30 C, -35 C and -40 C. The inclusion of 50 mM histidine in the glycine-containing formulation retarded NaCI crystallization. Consequently, the annealing time was increased 3-fold for such fonnulations in order to achieve an equivalent crystallinity.
However, the effect of 20 mM histidine on the crystallization of NaCI in the glycine-containing formulations was minimal. In freeze-drying studies, collapse of the lyophilized cake was observed visually in glycine-containing formulations containing 50 mM histidine. X-ray powder diffraction data is indicated a decrease in the crystallinity of NaCI in samples containing histidine.
In mannitol-containing formulations, typically 83% - 90% of the sodium chloride crystallized during cooling between -40*C and -50 C without the need for annealing. While inclusion of 20 mM histidine to the formulation suppressed NaCI crystallization during cooling, annealing resulted in approximately 40% crystallization of the NaCI.
Therefore, in formulations containing a crystallizable bulking agent, such as glycine or mannitol, and NaCI, the inclusion of histidine may decrease the extent of crystallizafion of NaCI. Although this could in some cases lead to the collapse of the cake which is formed during lyophilization, the use of relatively lower concentrations of histidine in such formulations can mitigate this effect.
Nonetheless, acceptable cakes have been formed with concentrations of histidine of 35 mM and 50 mM. Histidine may also be preferable to HEPES as a buffer in mannitol- and glycine-based formulations, as the use of HEPES has been observed to lower the Tg' to a greater extent than a similar amount of histidine.
Example 5 5 The physical cbaracteristics of a number of potential Factor VAI
fonrnulations, including seven candidate stabilizers and five bulking agents, were evaluated in another study. In addition to a bullcing agent and stabilizer, all formulations listed in Table 8 below (except for formulation 11) contained 10 mM Tris=HC1, 200 mM NaCI, 0.02% Tween-80, 4 mM CaC12 and were at pH 7Ø Fonmulation 11 contained 10 mM Tris*HC1, 0.02% Tween-80, and 4 mM CaCl2, also at pH
7Ø All pH measurements were performed at ambient tempexatuie.
}
Table 8 Sample Bulking Agent Protein I.D. Stabilizer 1 8% Mannitol 2% Sucrose 2 8% Mannitol 2% Trehalose 3 8% Iviannitol 2% Raffinose 4 8% Mannitol 2% Arginine 5 8% Mannitol 2% Lysine 6 8% Mannitol 2% Sorbitol 7 8% Mannitol 2% Glycerol
In accordance with an aspect of the present invention, there is provided a Factor VIII composition formulated without adding albumin to said composition, comprising the following formulation excipients in addition to Factor VIII: 2%
to 6% hydroxyethyl starch; 1% to 4% of a stabilizing agent selected from the group consisting of sucrose, trehalose, raffinose, and arginine; 1 mM to 5mM calcium salt; 100 mM to 300 mM NaCI; and a buffering agent for maintaining a pH of approximately between 6 and 8.
DETAILED DESCRIPTION OF THE INVENTION
Definitions As used herein, the terms below and variations thereof shall be defined as follows, unless otherwise indicated:
Factor VIII - The Factor VIII molecule exists naturally and in therapeutic preparations as a heterogeneous distribution of polypeptides arising from a single gene product (see, e.g., Anderson et al., Proc. Natl. Acad. Sci. USA, 83, 2979-2983, May 1986). The term "Factor VIII" as used herein refers to all such polypeptides, whether derived from blood plasma or produced through the use of recombinant DNA techniques. Commercially available examples of therapeutic preparations containing Factor VIII include those sold under the trade names of HEMOFIL M and RECOMBINATE (available from Baxter Healthcare Corporation, Deerfield, Illinois, U.S.A.). Other preparations currently in development comprise primarily a single subpopulation of Factor VIII molecules which lack the B domain portion of the molecule.
7a International Unit, IU- International Unit, or IU, is a unit of measurement of the blood coagulation activity (potency) of Factor VIII as measured by a standard assay, such as one of the following:
One stage assay. One stage assays are known to the art, such as that described in Lee, Martin L, et al., An Effect of Predilution on Potency Assays of Factor VIII Concentrates, Thrombosis Research (Pergamon Press Ltd.) 30, 511-519 (1983).
S
Chromogenic assay. Chromogenic assays maybe purchased commercially, sueh as the Coatest Factor VIII, available from Chromogenix AB, Molndal, Sweden.
Anneal The term anneal shall be used to indicate a step in the lyophilization process of a pharmaceutical preparation undergoing lyophilization, prior io the freeze-drying of the preparation, in which the temperature of the preptation is . ~\
raised from a]ower temperature to a higher temperature and then cooled again after a period of time.
Bulking Agent - For the purposes of this application, bulking agents are lhose chemical entities which provide structure to the "cake" or residual solid mass of a pharmaceutical preparation after it has been lyophilized and which protect it ag.ainst collapse. A crystallizable bulking agent shall mean a bulking agent as descn'bed herein which can be crystallizcd during lyophilization, other than sodium chloride. HES isnot included in this group of crystallizable bullang agents.
Freeze-drying, f freezin& lyophilizing -. 'Troeez.e-drying," unless otherwise indicated by the conteat in which it appens, shall be used to denote the portion of a lyophilization process in which the temperature of a pliarmaceutical preparation is raised in order to drive water out of the preparatioyn'. Th,e "freezing" steps of a lyophilization pmoess.are those steps which occur prior to the freez.e-drying stage. "Lyophilizing," unless othrawise indicated, shall refer 2S to the entire process of lyophilization, including both the Erees.ing steps and the freeze-drying steps.
Unless otherwise noted, percentage terms express weight/volume percentages and temperatures are in the Celsius scale.
Formulation Components The Factor VIII compositions of the present invention include bulking agents, stabilizing agents, buffering agents, sodium chloride, calcium salts, and, advantageously, other excipients. These excipients have been chosen in order to maximize the stability of Factor VIII in lyophilized preparations.. However, the Factor VIII compositions of the present invention exhibit stability in the liquid state as well.
The bulking agents used in the pn.sent formulations, which form the crystalline portion of the lyophilized product (except in the case of HES), are selected from the group consisting of mannitol, glycine, alanine, and hydroxyethyl starch (IM). Mannitol, glycine, or alanine are present in an amount of 4 - 10%, preferably 6- 9'/0, and more preferably about 8'/o. When HES is used as a bulking agent, it is present in an amount of 2 - 6%, preferably 3- 5%, and more preferably about 4%.
The stabilizing agents used in the formulations of the present invention are selected from the group consisting of sucrose, trehalose, raffinose, and arginine.
2o These agents are present in the formulations of the present invention in an atnount of between 1- 4%, preferably 2 - 3%, more preferably about 2%.
Sorbitol and glycerol were evaluated as possible stabilizers but were found to be poor stabilizers in the present formulations.
7,5 . Sodium chloride is included in the present formulations in an amount of 300 mM, preferably 150 - 250 mNrl, and most preferably about 225 mM. In one embodiment of the present invention, sodium chloride itself can be used without any of the aforementioned bulking agents, in which case it would be included in the fonnuiation in an amount of between 300 mM and 500 mM NaCI, preferably 350 to 450 mM NaCI, and more preferably about 400 mM NaCI.
In addition, buffers are present in these fonnulations, because it is believed that 5 the Factor VIII molecule can be adversely affected by pH shifts during lyophilization. The pH should preferably be maintained in the range of between 6 and 8 during lyophilization, and more preferably at a pH of about 7. The buffering agent can be any physiologically acceptable chemical entity or combination of chemical entities which have the capacity to act as buffers, 1 o including histidine, Tris, BIS-Tris Propane, PIPES, MOPS, HEPES, MES and ACES. The full cbeinical designations of these buffering agents is listed in Table I below. Typically; the buffering agent is included in a concentration of 10 - 50 mM. When histidine is added to the formulations, concentrations of over 20 mM and preferably about 25 mM are used, alone or in combination vvith other buffers such as Tris. Histidine is especially preferred for use in the compositions of the present invention, as descn'bed in greater detail below.
, ..= ' .
Table 1- Buffering Agents Tris tris-(hydroxymethyl)-aminomethane BIS-Tris Propane 1,3-bis-(tris-(hydroxy-methyl)methylamino) propane PIPES pipera2ine-N,N'-bis-(2-ethanesulfonic acid) MOPS 3-(N-morpholino) propanesulfonic acid HEPES N-2-hydroxyethyl-piperazine-N'-2-ethanesulfonic acid MES 2-(N-morpholino) ethanesulfonic acid ACES N-2-acetamido-2-aminoethanesulfonic acid In. order to preserve the activity of Factor VIII, it is inzportant that the formulations of the present invention also include calcium or another divalent cation able to interact with Factor VIII and maintain its activity, presumably by maintaining the association of the heavy and light chains of Factor VIII.
Between 1 mM and 5 mM of a calcium salt can be used, more preferably 3- 4 mM, and most preferably about 4 mM. The calcium salt is preferably calcium chloride, but can also be other calcium salts such as calcium gluconate, calcium glubionate, or calcium gluceptate.
The Factor VIII compositions of the present invention also preferably include a surfactant, preferably in an amount of 0.1 Jo or less, and more preferably in an amount of about 0.03%. The surfactant can, for example, be chosen from the group consisting of polysorbate 20, polysorbate 80, pluronic polyols, and Brij 35 (polyoxyethylene 231auryl ether). Several grades of pluronic polyols (sold under the trade name Pluronic, manufactured by the BASF Wyandotte Corporation) are available. These polyols, of diversified molecular weight (from 1,000 to over 16,000) and physicochemical properties have been used as surfactants. Pluronic F-38, of a molecular weight of 5,000 and Pluronic F-68, molecular weight 9,000, both contain (by weight) 80 per cent hydrophilic polyoxyethylene groups and 20 percent hydrophobic polyoxypropylene groups.
> = .
Tween-80, a commercial polysorbate, however, is preferred in the present formulations, in particular vegetable-derived Tween-80.
The Factor VIII formulations of thepresent invention also preferably include an antioxidant. The addition of antioxidants to the lyophilized formulations of the invention has been found to improve the stability of these formulations, and thus extend their shelf lives. The antioxidants used must be compatible for use with a pharmaceutical preparation, and in addition are preferably water soluble.
When adding antioxidants to a formulation, it is preferable to add such antioxidants as late in the process prior to lyophilization as possible, in order to avoid spontaneous oxidation of the antioxidant. Table 2 below lists suitable antioxidants, which are available commercially through companies such as Calbiochem and Sigma.
Table 2 - Antioxidants N-Acetyl-L-Cysteine / Homocysteine Glutathione -6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) Lipoic acid Methionine Sodium Thiosulfate Platinum Glycine-glycine-histidine (tripeptide) Butylatedbydroxytoluene (BITT) Of the foregoing antioxidants, glutathione is prefen=ed. Concentrations in the range of about 0.05 mg/mI to more than 1.0 mgtml have all been found to ',. enhance the stability of Factor VIII compositions, and it is believed that higher concentrations would also be useful (up to the point of any toxic effects or adverse manufacturing effects, such as a depression of the glass transition temperature of the lyophilized product).
It has been found in particular that the combination of histidine and glutathione produces synergistically beneficial effects on the stability of Factor VIII
compositions. Histidine, while acting as a buffer, can also act as a metal chelator. To the extent that Factor VIII inactivation is caused by metal-induced oxidation, histidine can therefore act to stabilize Factor VIII by binding such oxidizing metal ions. It is believed that by binding these metals, the glutathione (or indeed any other antioxidant present) is thereby able to provide fnrther antioxidative protection, since the oxidative effect of the metal ions bound by.
the histidine has been contained.
Other chelating agents might also be used in the compositions of the present invention. Such agents should preferably bind metals such as copper and iron with greater affinity than calcium, if a calcium salt is being used in the composition. One such chelator is deferoxamine, a chelating agent that facilitates the removal of AI++ and iron. Deferoxamine Mesylate, C25H48N608*CH4O3S, can be obtained from Sigma (Sigma Prod. No.
D9533). It is an aluminum and iron(II) chelator which chelates iron (as a 1:1 chelate complex) only in the +3 oxidation state, not +2 oxidation state, and ean also bind manganese ion and other metals. Deferoxamine can be used advantageously in an amount of 0.25 mg/1.
The Factor VIII used in the present formulations can be either highly purified human plasma-derived Factor VIII or more preferably can be recombinantly =~
produced Factor VIII. Recombinant Factor vIII can be produced by Chinese hamster ovary (CHO) cells transfected with a vector canying a DNA sequence coding for the Factor VIII molecule. Methods for creating such transfected CHO
cells are described, inter alia, in U.S. Patent No. 4,757,006 to Toole, Jr., though alternative methods are also known to the art (see, e.g., U.S. Patent No.
4,868,112, also to Toole, Jr., and PCT International Application WO A-91/09122). The methods used to culture sucb CHO cells to produce Factor VIII
are also lcnown to the art, for example in European Patent Application No. 0 218 to Genetics Institute, entitled "Improved method for producing Factor VIII:C-type proteins." Recombinant Factor VIII can, however, also be produced in other cell lines, such as baby hamster kidney (BHK) cells. The Factor VIII
molecule itself, if recombinantly produced, can be either full-length Factor VIII
or a deletion derivative thereof, such as a B domain-deleted Factor VIII
molecule.
While the Factor VIII compositions descnbed in this application can be lyophilized and reconstituted in the indicated concentrations, one of skill in the art will understand that these preparations can also be reconstituted in more dilute form. For example, a preparation according the present invention which is lyophilized and/or normally reconstituted in 2 ml of solution can also be reconstituted in a larger volume of diluent, such as 5 ml. This is particularly appropriate when the Factor VIII preparation is being injected into a patient immediately, since in this case the Factor VIII is less likely to lose activity, -which may occur more rapidly in more dilute solutions of Factor VIII.
Formulation and LyopbiGzation Development In order to achieve maximal stability, the Factor VIII compositions of the present invention are preferably lyophilized. During lyophilization, Factor VIII
is converted from being in an aqueous phase to being in an asnorphous solid t, phase, which is thought to protect the protein from chemical and/or confonnational instability. The lyophilized preparation not only contains an amorphous phase, but also includes a coiinponent wluch crystallizes during lyophilization. This is thought to allow the rapid lyophilization of the Factor VIII composition and the fonnation.of a more elegant cake (that is, a cake with minimal shrinkage from the sides of the container in which it was lyophilized).
In the formulations of the present invention, the stabilizing agents have been selected to exist primarily in an amorphous phase of the lyophilized product, while the bulking agents (except HES) have been selected to crystallize during freezing.
WO 00/48635 PCT/[1S00/40068 Both the Factor VIII and the stabilizer are preferably dispersed in the amorphous phase of the lyophilized cake. The mass of the stabilizer is also preferably large compared to the other excipients in the amorphous form. In addition, the apparent glass transition temperature (Ts') of the amorphous phase 5 is preferably relatively high during freeze-drying, and the glass transition temperature (Tg) of the solid is likewise preferably high during storage.
Crystallization of sodium chloride in the product was found to be desirable, since amorphous sodium cbloride will depm,ss the T.' of the amorphous phase.
10 In order to avoid the collapse of the cake of a particular composition, primary drying is preferably carried out at a product temperature below the apparent glass transition temperature of the freeze concentrate. . An increase in drying time may also be required to offset a decrease in Ts'. Further infonnation on lyophilization may be found in Carpenter, J.F. and Chang, B.S., Lyophi7izatinn 15 ofProtein Pharmaceuticals, BiotechnoloQV and Biopharmaccutical Manufacturing, Processing and Preservation, K.E. Avis and V.L. Wu, eds.
(Buffalo Grove, IL: Interpharm Press, Inc.), pp. 199-264 (1996).
;
Example 1 The effects of the concentration of Factor VIII and of the addition of a stabilizer on the recovery of Factor VIII were investigated in several studies. These studies were perfonmed using mannitol as a model bulking agent and sucrose as a model stabilizer. The three sample formulations described in Table 3 below were used in these studies. All formulations used in these studies included 10 mM Tris, 200 mM NaCI, 8% mannitol, 4 mM CaCl2, and 0.02% Tween-80 and were conducted at pH 7Ø
Table 3 Sample Initial Factor VIII Sucrose %
I.D. (ILT/m!) IA. 600 1iese samples were lyophilized using the freeze-drying cycle shown in Table 4 below in order to maintain a product temperature below the apparent glass s transition temperature (T,'). Differential scanning calorimetric (DSC) studies indicated the presence of a transition at approximately -40 C in the mannitol formulations. In order to maintain a product temperature below this value, the shelf temperature was set to -32 C during primary drying. Primary drying under these conditions was perfon:ned for about 55 hours, with a total cycle time io of about 80 hours.
,., Table 4 Freezing/Processing Description Metbod I Cool to +5 C;
(Freezing) Cool to -5 C at 1 C/minute, hold for 20 minutes;
Cool to -20 + 5 C at 1 C/minute; hold for 1 hour (up to 3 hours);
Cool to -45 C at 0.5 C/minute, hold for 1 hour.
II Freeze per method I
Hold at -35 C for 48 hours.
m Freeze per method I
Hold at -35 C for 48 hours;
Hold at 20 C for 48 hours.
IV Shelf -32 C during primary drying for (Freeze-drying) about 55 hours (up to 100 hours);
Product < -40 C during primary drying;
Ramp from -32 C to +40 C at 0.2 C/minute;
Shelf +40 C during secondary drying for 3 hours.
-, , The Factor VIII activity of these samples, as determined by the one-stage clotting assay, was compared against a control held at -45 C. The assay results are shown in Table 5 below.
Table 5 Processing % Loss in Factor VIII Activity During Each Step Method Formulation IA Formulation IB Formulation IC
(600 IU/ml) (60 IU/ml) (60 IU/ml, 2%
Sucrose) 1 6.7 37.5 41.7 II. 2.0 9.3 3.9 III 7.3 11.6 5.0 N 20.0 24.2 18.3 (Lyophilization) These results indicate that protein concentration has an effect on the recovery of Factor VIII during freezing. Formulations containing 60 IU/ml lost approximately 37 lr-42 !0 of the initial Factor VIII activity during the freezing step, while 6.7% of Factor VIII activity was lost for the formulation containing 600 IU/ml. These results indicate that a higher protein concentration has a protective effect during ficezzing. Although sucrose provided some protection to the Factor VIII during the intermediate tonpera'ture holds as well as during freeze-drying, it failed to protect the protein during the initial freezing step.
Example 2 Following the development of the lyophilization process outlined in Example 1, further optimization of this process was undertaken. It has been found that a lyophilized composition having a higher glass transition temperature, (and, theoretically, better Factor VIII stability) can be produced by: (1) lowering the freezing temperature initially to -45 C or lower (such as down to about -50 C
or -55 C); (2) raising the temperature to -20 C or -22 C (+5 C); and then (3) lowering the temperature again to -45 C or lower. The temperature is lowered or raised, as the case may be, at a rate of between about 0.5 C and about 1.0 C
per minute. Once the desired temperature is reached, the composition is held at that temperature for between I and 3 hours. This improved freezing cycle is shown in Table 6 below.
Table 6 Freezing Method Description I Cool to +5 C;
Cool to -5 C at 0.5-1 Clminut.e, hold for 20 niinutes;
Cool to between-55 C and -45 C at 0.5-1 C/minute, hold for about 1 hour, Warm to -22 C ( 5 C) at 0.5-1 CJminute, bold for I to 3 hours;
Cool tio -45 C at 0.5-1 C/minute, hold for about 1 hour.
Unless otherwise indicated, the temperatures referred to in this example and in other examples refer to the shelf temperature of the lyophilizer and not to the temperature of the product per se. Following the improved freezing cycle, the remainder of the lyophilization process can be conducted as outlined in Example 1 above, or otherwise as described fiuther herein or as determined by one of skill in the art.
This improved lyophilization process was found to be useful for fonnulations which include glycine as the bulking agent as well as those which use mannitol.
It is fnrther believed to have applicability to formulations which maice use of the other bullcing agents of the present invention as well.
WO 00/48635 PCT/US00l40068 Example 3 It is believed that in order to produce a freeze-dried product with acceptable cake appearance and glass transition temperature, the bulking agent of lyophilized pharmaceutical preparations which contain sodium chloride, such as 5 glycine or mannitol, may need to be crystallized. The following improved lyophilization process for crystallizable bulking agents was therefore developed.
Table 7a - Freezing Steps Process Step Temperature Duration of Step Initial freezing -40 C or less 1 hour First annealing between -23 C and 27 C 3 hours Second freeyin,g -55 C 1 hour Second annealing -36 C 4 bours Third freezing -50 C 1 hour ' ~ . Table 7b - Freeze-Drying Steps Process Step Temperature Duration of Step Primary Drying -35 C up to 100 hours Secondary Drying: First step 40 C 3 hours }
Secondary Drying: Second step 45 C 3 hours Secondary Drying: Third step 50 C 3 hours In the freezing steps, changes in the temperatures occurred at a rate of between about 0.5 C/minute and 1 C/minute.' It is believed that steps of longer duration would also be effective.
Prior to the first freezing step, the temperature is brought to between about and 8 C for about one hour for the purpose of bringing all the vials to approximately the same temperature. After this the lyophilizer is cooled to -C. The first freezing step should be perfonned at a temperature less than -5 30 C, preferably below -35 C, and more preferably at about -40 C: Following this, the first annealing step should occur at a temperature of between -30 C
and -19 C, more preferably either between about 25 C and -28 C (if glycine is the bulking agent) or between -21 C and 24 C (if mannitol is the bulking agetit), with the temperatures of 23 C and 26 C being most preferred, at which temperatures it is believed that the crystallizable bulking agents crystallize, at least in part. However, the lower range around -27 C is not recommended for formulations containing mannitol and arginine. This step is preferably carried out for about 3 hours.
Following the first annealing step, the temperature is lowered, preferably to less than about -50 C and more preferably to less than -55"C, for about 1 hour. It is believed that the sodium chloride in the preparation nucleates at this time.
During the second annealing step, the temperature of the pharmaceutical preparation is raised to between about --30 C and -39 C, and preferably to about -33 C for rnannitol-containing compositions and -36 C for glycine-containing compositions. It is believed that NaCI crystal growth occurs at this time, at least in part. This step is preferably conducted for about 4 hours. Following this, the temperature of the lyophilizer is reduced to about -50 C, preferably for about hour in order to reduce the temperature of the preparation.
In the freeze-drying steps which follow, changes in temperature occurred at a rate of between about 0.1 C/minute and 0.5 C/minute. After reducing the pressure in the lyophilizer to about'65 mTorr, the temperature is raised to between about -32 C and -35 C for primary drying. Ice crystals in the preparation will sublimate at this temperature. This step is performed for up to about 100 hours, or until most of the ice has been sublimated from the preparation. The point at which most of the ice has sublimated can be determined, for example, using a dewpoint sensor, which indicates the end of the sublimation of ice when the readings decrease (the point of inflection).
Following primary drying, the temperature is raised to +40 C, preferably at a rate of 0.2 Clminute, to initiate secondary drying to remove fiuther water from the preparation. This temperature is preferably maintained for about three hours. Second and third secondary drying steps follow this first step, where the temperature is raised to about +45 C for about three hours and then to about +50 C for three more hours in order to reduce the moisture in the lyophilized cake to less than 2% (w/w).
Eiample 4 Further studies were performed to examine specifically the effect of histidine on lyophilized Factor VIII compositions containing glycine or mannitol as bulking agents. Non-reversing beat flow (Modulated DSC, mDSC) was used to detect the crystallization of these bulking agents during cooling. Both the temperature of crystallization and the total heat of crystallization were determined from the crystallization exotherm. The appearance of the NaCI eutectic melt endotherm during warming was used to detect NaCI crystalliza6on. In mDSC, the extent of crystallization was determined as the ratio of the enthalpy of melting of the formulation to the enthalpy of melting of pure NaCI solution by using the total heat flow signal. In addition, X-ray diffraction analyses were performed in order to determine the extent of crystallization in the lyophilized formulations.
WO 00/48635' PCT/US00/40068 While histidine concentrations less than 20 mM did not significantly impact the crystallization of glycine, 50 mM histidine reduced the extent of glycine crystallization. Well-defined NaCI crystallization exotherms were not observed during cooling of formulations containing glycine. However, eutectic melting endotherms during heating indicated that NaCI was crystallized (> 50%) after cooling lower than 50 C and annealing at 30 C, -35 C and -40 C. The inclusion of 50 mM histidine in the glycine-containing formulation retarded NaCI crystallization. Consequently, the annealing time was increased 3-fold for such fonnulations in order to achieve an equivalent crystallinity.
However, the effect of 20 mM histidine on the crystallization of NaCI in the glycine-containing formulations was minimal. In freeze-drying studies, collapse of the lyophilized cake was observed visually in glycine-containing formulations containing 50 mM histidine. X-ray powder diffraction data is indicated a decrease in the crystallinity of NaCI in samples containing histidine.
In mannitol-containing formulations, typically 83% - 90% of the sodium chloride crystallized during cooling between -40*C and -50 C without the need for annealing. While inclusion of 20 mM histidine to the formulation suppressed NaCI crystallization during cooling, annealing resulted in approximately 40% crystallization of the NaCI.
Therefore, in formulations containing a crystallizable bulking agent, such as glycine or mannitol, and NaCI, the inclusion of histidine may decrease the extent of crystallizafion of NaCI. Although this could in some cases lead to the collapse of the cake which is formed during lyophilization, the use of relatively lower concentrations of histidine in such formulations can mitigate this effect.
Nonetheless, acceptable cakes have been formed with concentrations of histidine of 35 mM and 50 mM. Histidine may also be preferable to HEPES as a buffer in mannitol- and glycine-based formulations, as the use of HEPES has been observed to lower the Tg' to a greater extent than a similar amount of histidine.
Example 5 5 The physical cbaracteristics of a number of potential Factor VAI
fonrnulations, including seven candidate stabilizers and five bulking agents, were evaluated in another study. In addition to a bullcing agent and stabilizer, all formulations listed in Table 8 below (except for formulation 11) contained 10 mM Tris=HC1, 200 mM NaCI, 0.02% Tween-80, 4 mM CaC12 and were at pH 7Ø Fonmulation 11 contained 10 mM Tris*HC1, 0.02% Tween-80, and 4 mM CaCl2, also at pH
7Ø All pH measurements were performed at ambient tempexatuie.
}
Table 8 Sample Bulking Agent Protein I.D. Stabilizer 1 8% Mannitol 2% Sucrose 2 8% Mannitol 2% Trehalose 3 8% Iviannitol 2% Raffinose 4 8% Mannitol 2% Arginine 5 8% Mannitol 2% Lysine 6 8% Mannitol 2% Sorbitol 7 8% Mannitol 2% Glycerol
8 4% Hydroxyethyl 2% Sucrose Starch
9 8% Glycine 2% Sucrose ~ . . .
10 8% Glycine 2% Trebalose
11 400 nilvl NaCI 2% Sucrose
12 8% Alanine 2% Sucrose Collapse temperature measurements by freeze-dry microscopy and thermal 5 transition measurements by DSC were used to predict freeze-drying behavior.
DSC, X-ray powder diffraction and polarized light microscopy were also used to determine the crystallinity of the lyophilized samples. The reconstitution time and the appearance of the samples were also evaluated. The results of all of these measurements are summarizcd in Table 9 below.
Table 9 Sample Tw T, Tg Reconstitution Water I.D. (oC) (oC) ( C) (seconds) Content Appearance (%) 1 -14 -10 54 64 n/c Elegant 2 -20 -15 53 62 1.4 Top partially collapsed 3 -15 -10 54 77 1.7 Elegant 4 - - - - Partial collapse - - - - Collapsed 6 n/c n/c < 10 C' 63 0.6 Elegant 7 - - < 10 C' - - Elegant 8 - - 86 49 0.7 Elegant but shrunk from sides 9 - 54 22 0.8 Elegant - - 63 18 - Elegant 11 - - 66 11 0.4 Elegant (layer on bottom) 12 - - - 57 0.5 Elegant Sorbitol and Glycerol have glass transitions at < 10 C. The DSC scan range did not include temperatures in this range.
n/c = not clear 5 T1. = Temperature at which partial collapse occurs in the freeze-dry microscope T. = Temperature at which total collapse occurs in the freeze-dry microscope Tg = Glass transition temperature io With the exception of mannitol:lysine, all of the formulations appeared to have adequate physical appearance. Lysine interfered with the crystallization of both WO 00/48635 PC'i'/US00/40068 mannitol and glycine, which caused a depression in the glass transition temperature and a collapse of the lyophilized cake.
Example 6 The Factor VIII compositions descn-bed in Table 8 above were placed in storage at -70 C, 25 C, 40 C, and 50 C for varying lengths of time in order to evaluate their stability. Factor VIII activity levels were evaluated after 2 weeks, I
month, 2 months, and 3 months, and the results are sunimarized in Table i 0 below. Two of the samples, one employing mannitol as the bulking agent and sorbitol as the stabilizer, and the other employing mannitol as the bulking agent lo and glycerol as the stabilizer, exhibited poor stability. The remaining formulations all exhibited the ability to stabilize Factor VIII.
Table 10 Formulation Temperature % of initial at month Description ( C) 0 0.5 1 2 3 ~'. Glycine:Sucrose -70 100.00 97.43 101.71 99.89 97.97 25 100.00 85.44 40 100.00 79.87 71.52 63.06 50 100.00 76.34 67.99 52.14 47.64 Glycine:Trehalose -70 100.00 89.22 96.00 95.90 94.64 25 100.00 83.17 40 100.00 79.93 72.42 68.03 50 100.00 80.97 64.28 57.60 50.92 Mannitol:Trehalose -70 100.00 91.32 97.72 96.10 98.26 25 100.00 85:79 40 100.00 82.54 70.72 59.44 50 100.00 66.16 65.51 48.81 52.06 Mannitol:Sucrose -70 100.00 100.45 100.56 105.47 99.22 25 100.00 87.04 40 100.00 85.59 80.78 55.42 50 100.00 81.68 75.53 57.88 43.46 WO 00/48635 PCTfUS00/40068 Mannitol:Arginine -70 1100.00 102.26 105.53 103.72 105.08 25 100.00 95.15 40 100.00 91.53 80.93 69.19 50 100.00 82.28 68.06 56.32 45.94 Mannitol:Raffinose -70 100.00 93.88 98.41 100.68 103.62 25 100.00 83.13 40 100.00 81.09 73.61 67.16 50 100.00 71.69 68.52 54.25 47.11 Mannitol:Glycerol -70 Mannitol:Sorbitol -70 100.00 104.06 25 100.00 40 100.00 50 100.00 32.73 HES:Sucrose -70 100.00 102.74 103.03 100.90 25 100.00 40 100.00 76.89 77.47 50 100.00 71.47 67.40 30.02 NaCI:Sucrose -70 100.00 88.54 88.44 95.58 25 100.00 40 100.00 71.56 58.30 50 100.00 52.71 37.90 30.34 = , Alanine:Sucrose -70 100.00 109.78 109.67 108.96 25 100.00 40 100.00 92.99 73.03 50 100.00 83.25 74.91 57.65 Glycine:Raffinose -70 100.00 11 -1.57 114.51 105.25 25 100.00 40 100.00 89.20 82.10 50 100.00 93.21 72.22 53.24 Example 7 5 Based on the information developed during the studies descrbed in Examples 5 and 6, it was decided that candidate formulations having the excipients shown in Table 1 I below would be further developed.
Table 11 Excipient Concentration mannitol or glycine 6-9%
arginine or trehalose 1-3%
tween 80 = 0.005-0.04%
NaCI 200-250 mM
CaC12 3-5 mM
TRIS 20-30 mM
histidine or HEPES 10-50 nilvl glutathione 0.15-0.25 mg/ml Based on these parameters, the following specific formulations were developed:
Table 12 , Formulation #1 Formulation #2 Formulation #3 10mM HEPES l OmM REPES I OniM HEPES
20mM Tris .20mM Tris 20mM Tris 225mM NaCI 225mM NaCI 225mM NaCI
0.03% (v/v) Tween-80 0.03% (v/v) Tween-80 0.03% (v/v) Tween-80 8% (w/v) mannitol 8% (w/v) glycine 8% (w/v) mannitol 2% (w/v) trehalose 2% (w/v) trehalose 2% (w/v) arginine 02 mg/ml reduced 0.2 mg/ml reduced 0.2 mgfml reduced }
glutathione glutathione glutatbione 4 mM CaC12 4 mM CaC12 4 mM CaC1Z
w0 00/48635 PCTIUSOO/40068 Formulation #4 Formulation #5 25mM histidine 25mM histidine 20mM Tris 20mM Tris 225mM NaCI 225mM NaCI
0.03% (v/v) Tween-80 0.03% (v/v) Tween-80 8% (w/v) mannitol 8% (w/v) glycine 2% (w/v) trehalose 2% (w/v) trehalose 02 rng/ml reduced 0.2 mgfml reduced glutathione glutathione 4 mM CaC12 4 mM CaC12 , =~ .
.f .
, . .
DSC, X-ray powder diffraction and polarized light microscopy were also used to determine the crystallinity of the lyophilized samples. The reconstitution time and the appearance of the samples were also evaluated. The results of all of these measurements are summarizcd in Table 9 below.
Table 9 Sample Tw T, Tg Reconstitution Water I.D. (oC) (oC) ( C) (seconds) Content Appearance (%) 1 -14 -10 54 64 n/c Elegant 2 -20 -15 53 62 1.4 Top partially collapsed 3 -15 -10 54 77 1.7 Elegant 4 - - - - Partial collapse - - - - Collapsed 6 n/c n/c < 10 C' 63 0.6 Elegant 7 - - < 10 C' - - Elegant 8 - - 86 49 0.7 Elegant but shrunk from sides 9 - 54 22 0.8 Elegant - - 63 18 - Elegant 11 - - 66 11 0.4 Elegant (layer on bottom) 12 - - - 57 0.5 Elegant Sorbitol and Glycerol have glass transitions at < 10 C. The DSC scan range did not include temperatures in this range.
n/c = not clear 5 T1. = Temperature at which partial collapse occurs in the freeze-dry microscope T. = Temperature at which total collapse occurs in the freeze-dry microscope Tg = Glass transition temperature io With the exception of mannitol:lysine, all of the formulations appeared to have adequate physical appearance. Lysine interfered with the crystallization of both WO 00/48635 PC'i'/US00/40068 mannitol and glycine, which caused a depression in the glass transition temperature and a collapse of the lyophilized cake.
Example 6 The Factor VIII compositions descn-bed in Table 8 above were placed in storage at -70 C, 25 C, 40 C, and 50 C for varying lengths of time in order to evaluate their stability. Factor VIII activity levels were evaluated after 2 weeks, I
month, 2 months, and 3 months, and the results are sunimarized in Table i 0 below. Two of the samples, one employing mannitol as the bulking agent and sorbitol as the stabilizer, and the other employing mannitol as the bulking agent lo and glycerol as the stabilizer, exhibited poor stability. The remaining formulations all exhibited the ability to stabilize Factor VIII.
Table 10 Formulation Temperature % of initial at month Description ( C) 0 0.5 1 2 3 ~'. Glycine:Sucrose -70 100.00 97.43 101.71 99.89 97.97 25 100.00 85.44 40 100.00 79.87 71.52 63.06 50 100.00 76.34 67.99 52.14 47.64 Glycine:Trehalose -70 100.00 89.22 96.00 95.90 94.64 25 100.00 83.17 40 100.00 79.93 72.42 68.03 50 100.00 80.97 64.28 57.60 50.92 Mannitol:Trehalose -70 100.00 91.32 97.72 96.10 98.26 25 100.00 85:79 40 100.00 82.54 70.72 59.44 50 100.00 66.16 65.51 48.81 52.06 Mannitol:Sucrose -70 100.00 100.45 100.56 105.47 99.22 25 100.00 87.04 40 100.00 85.59 80.78 55.42 50 100.00 81.68 75.53 57.88 43.46 WO 00/48635 PCTfUS00/40068 Mannitol:Arginine -70 1100.00 102.26 105.53 103.72 105.08 25 100.00 95.15 40 100.00 91.53 80.93 69.19 50 100.00 82.28 68.06 56.32 45.94 Mannitol:Raffinose -70 100.00 93.88 98.41 100.68 103.62 25 100.00 83.13 40 100.00 81.09 73.61 67.16 50 100.00 71.69 68.52 54.25 47.11 Mannitol:Glycerol -70 Mannitol:Sorbitol -70 100.00 104.06 25 100.00 40 100.00 50 100.00 32.73 HES:Sucrose -70 100.00 102.74 103.03 100.90 25 100.00 40 100.00 76.89 77.47 50 100.00 71.47 67.40 30.02 NaCI:Sucrose -70 100.00 88.54 88.44 95.58 25 100.00 40 100.00 71.56 58.30 50 100.00 52.71 37.90 30.34 = , Alanine:Sucrose -70 100.00 109.78 109.67 108.96 25 100.00 40 100.00 92.99 73.03 50 100.00 83.25 74.91 57.65 Glycine:Raffinose -70 100.00 11 -1.57 114.51 105.25 25 100.00 40 100.00 89.20 82.10 50 100.00 93.21 72.22 53.24 Example 7 5 Based on the information developed during the studies descrbed in Examples 5 and 6, it was decided that candidate formulations having the excipients shown in Table 1 I below would be further developed.
Table 11 Excipient Concentration mannitol or glycine 6-9%
arginine or trehalose 1-3%
tween 80 = 0.005-0.04%
NaCI 200-250 mM
CaC12 3-5 mM
TRIS 20-30 mM
histidine or HEPES 10-50 nilvl glutathione 0.15-0.25 mg/ml Based on these parameters, the following specific formulations were developed:
Table 12 , Formulation #1 Formulation #2 Formulation #3 10mM HEPES l OmM REPES I OniM HEPES
20mM Tris .20mM Tris 20mM Tris 225mM NaCI 225mM NaCI 225mM NaCI
0.03% (v/v) Tween-80 0.03% (v/v) Tween-80 0.03% (v/v) Tween-80 8% (w/v) mannitol 8% (w/v) glycine 8% (w/v) mannitol 2% (w/v) trehalose 2% (w/v) trehalose 2% (w/v) arginine 02 mg/ml reduced 0.2 mg/ml reduced 0.2 mgfml reduced }
glutathione glutathione glutatbione 4 mM CaC12 4 mM CaC12 4 mM CaC1Z
w0 00/48635 PCTIUSOO/40068 Formulation #4 Formulation #5 25mM histidine 25mM histidine 20mM Tris 20mM Tris 225mM NaCI 225mM NaCI
0.03% (v/v) Tween-80 0.03% (v/v) Tween-80 8% (w/v) mannitol 8% (w/v) glycine 2% (w/v) trehalose 2% (w/v) trehalose 02 rng/ml reduced 0.2 mgfml reduced glutathione glutathione 4 mM CaC12 4 mM CaC12 , =~ .
.f .
, . .
Claims (8)
1. A Factor VIII composition formulated without adding albumin to said composition, comprising the following formulation excipients in addition to Factor VIII:
2% to 6% hydroxyethyl starch;
1% to 4% of a stabilizing agent selected from the group consisting of sucrose, trehalose, raffinose, and arginine;
1 mM to 5mM calcium salt;
100 mM to 300 mM NaCl; and a buffering agent for maintaining a pH of approximately between 6 and 8.
2% to 6% hydroxyethyl starch;
1% to 4% of a stabilizing agent selected from the group consisting of sucrose, trehalose, raffinose, and arginine;
1 mM to 5mM calcium salt;
100 mM to 300 mM NaCl; and a buffering agent for maintaining a pH of approximately between 6 and 8.
2. The Factor VIII composition of claim 1, comprising about 4%
hydroxyethyl starch.
hydroxyethyl starch.
3. The Factor VIII composition according to claims 1 to 2, comprising about 200 mM NaCl.
4. The Factor VIII composition according to claims 1 to 3, wherein said stabilizing agent is present in an amount of about 2%.
5. The Factor VIII composition according to claims 1 to 4, wherein said stabilizing agent is sucrose.
6. The Factor VIII composition according to claims 1 to 4, wherein said stabilizing agent is arginine.
7. The Factor VIII composition according to claims 1 to 4, wherein said stabilizing agent is trehalose.
8. Use of a Factor VIII composition according to any one of claims 1 to 7 for the preparation of a medicament for the treatment of hemophilia.
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US25527999A | 1999-02-22 | 1999-02-22 | |
US09/255,279 | 1999-02-22 | ||
US45275299A | 1999-12-01 | 1999-12-01 | |
US09/452,752 | 1999-12-01 | ||
CA2362927A CA2362927C (en) | 1999-02-22 | 2000-02-22 | Novel albumin-free factor viii formulations |
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CA2362927A Division CA2362927C (en) | 1999-02-22 | 2000-02-22 | Novel albumin-free factor viii formulations |
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CA2634663A1 CA2634663A1 (en) | 2000-08-24 |
CA2634663C true CA2634663C (en) | 2009-08-25 |
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CA002634663A Expired - Lifetime CA2634663C (en) | 1999-02-22 | 2000-02-22 | Novel albumin-free factor viii formulations |
CA2634664A Expired - Lifetime CA2634664C (en) | 1999-02-22 | 2000-02-22 | Novel albumin-free factor viii formulations |
CA002634674A Abandoned CA2634674A1 (en) | 1999-02-22 | 2000-02-22 | Novel albumin-free factor viii formulations |
CA2362927A Expired - Lifetime CA2362927C (en) | 1999-02-22 | 2000-02-22 | Novel albumin-free factor viii formulations |
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CA2634664A Expired - Lifetime CA2634664C (en) | 1999-02-22 | 2000-02-22 | Novel albumin-free factor viii formulations |
CA002634674A Abandoned CA2634674A1 (en) | 1999-02-22 | 2000-02-22 | Novel albumin-free factor viii formulations |
CA2362927A Expired - Lifetime CA2362927C (en) | 1999-02-22 | 2000-02-22 | Novel albumin-free factor viii formulations |
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US (8) | US6586573B1 (en) |
EP (5) | EP1820516B1 (en) |
JP (9) | JP5149470B2 (en) |
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AT (1) | ATE365052T1 (en) |
AU (1) | AU777972B2 (en) |
BR (1) | BR0008405B1 (en) |
CA (4) | CA2634663C (en) |
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CZ (3) | CZ307322B6 (en) |
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Families Citing this family (126)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7244824B2 (en) * | 1995-01-19 | 2007-07-17 | Quadrant Drug Delivery Limited | Dried blood factor composition comprising trehalose |
US7253262B2 (en) * | 1995-01-19 | 2007-08-07 | Quandrant Drug Delivery Limited | Dried blood factor composition comprising trehalose |
GB9501040D0 (en) * | 1995-01-19 | 1995-03-08 | Quadrant Holdings Cambridge | Dried composition |
US6214054B1 (en) | 1998-09-21 | 2001-04-10 | Edwards Lifesciences Corporation | Method for fixation of biological tissues having mitigated propensity for post-implantation calcification and thrombosis and bioprosthetic devices prepared thereby |
PT2193809E (en) * | 1999-02-22 | 2015-08-24 | Baxter Int | Albumin-free factor viii formulations |
US6830917B2 (en) * | 2001-12-10 | 2004-12-14 | Baxter Healthcare S.A. | Method of isolation and purification of trypsin from pronase protease and use thereof |
BR0215216A (en) | 2001-12-21 | 2004-11-16 | Novo Nordisk Healthcare Ag | Liquid aqueous composition, method for preparing a liquid aqueous composition of a factor vii polypeptide, use of a liquid aqueous composition, and method for treating a factor vii response syndrome |
KR20040065278A (en) * | 2001-12-21 | 2004-07-21 | 노보 노르디스크 에이/에스 | Liquid composition of modified factor vii polypeptides |
US6878168B2 (en) | 2002-01-03 | 2005-04-12 | Edwards Lifesciences Corporation | Treatment of bioprosthetic tissues to mitigate post implantation calcification |
US6982080B2 (en) * | 2002-03-15 | 2006-01-03 | Wyeth | Hydroxyethyl starch—containing polypeptide compositions |
GB0207092D0 (en) * | 2002-03-26 | 2002-05-08 | Sod Conseils Rech Applic | Stable pharmaceutical composition containing factor VIII |
US20040009918A1 (en) * | 2002-05-03 | 2004-01-15 | Hanne Nedergaard | Stabilised solid compositions of modified factor VII |
KR101212025B1 (en) * | 2002-06-21 | 2013-01-09 | 노보 노르디스크 헬스 케어 악티엔게젤샤프트 | Stabilised solid compositions of factor ⅶ polypeptides |
GB0304636D0 (en) * | 2003-02-28 | 2003-04-02 | Britannia Pharmaceuticals Ltd | Pharmaceutical composition for nasal delivery |
CA2518327A1 (en) * | 2003-03-18 | 2004-09-30 | Novo Nordisk Health Care Ag | Liquid, aqueous, pharmaceutical compositions of factor vii polypeptides |
US7897734B2 (en) * | 2003-03-26 | 2011-03-01 | Novo Nordisk Healthcare Ag | Method for the production of proteins |
AU2004241698A1 (en) * | 2003-05-23 | 2004-12-02 | Novo Nordisk Health Care Ag | Protein stabilization in solution |
ES2382157T3 (en) * | 2003-06-25 | 2012-06-05 | Novo Nordisk Health Care Ag | Liquid composition of factor VII polypeptides |
ATE446768T1 (en) * | 2003-07-01 | 2009-11-15 | Novo Nordisk Healthcare Ag | LIQUID AQUEOUS PHARMACEUTICAL COMPOSITION OF FACTOR VII POLYPEPTIDES |
DE10333317A1 (en) * | 2003-07-22 | 2005-02-17 | Biotecon Therapeutics Gmbh | Formulation for protein medicines without the addition of human serum albumin (HSA) |
CA2531988C (en) * | 2003-08-05 | 2016-06-28 | Novo Nordisk A/S | Novel insulin derivatives |
KR100560697B1 (en) * | 2003-08-06 | 2006-03-16 | 씨제이 주식회사 | Formulation of albumin-free erythropoietin |
EP1656158B1 (en) | 2003-08-14 | 2016-03-09 | Novo Nordisk Health Care AG | Liquid, aqueous pharmaceutical composition of factor vii polypeptides |
US8541002B2 (en) * | 2003-09-12 | 2013-09-24 | Agenus Inc. | Vaccine for treatment and prevention of herpes simplex virus infection |
MXPA06006886A (en) * | 2003-12-19 | 2006-09-04 | Novo Nordisk Healthcare Ag | Stabilised compositions of factor vii polypeptides. |
GB0404586D0 (en) * | 2004-03-01 | 2004-04-07 | Britannia Pharmaceuticals Ltd | Improvements in or relating to organic materials |
WO2005089712A1 (en) * | 2004-03-04 | 2005-09-29 | Wyeth | Lyophilization method to improve excipient crystallization |
CA2557061C (en) * | 2004-03-19 | 2013-08-20 | Baxter International Inc. | Factor ixa for the treatment of bleeding disorders |
US7319032B2 (en) * | 2004-04-22 | 2008-01-15 | Medtox | Non-sugar sweeteners for use in test devices |
KR100624013B1 (en) * | 2004-06-25 | 2006-09-19 | 주식회사 녹십자홀딩스 | Pharmaceutical preparation of recombinant factor ? lyophilized without albumin as a stabilizer |
PT1835938E (en) * | 2004-12-27 | 2013-11-06 | Baxter Int | Polymer-von willebrand factor-conjugates |
FR2881139A1 (en) * | 2005-01-26 | 2006-07-28 | Agronomique Inst Nat Rech | Liquid for lyophilization of proteins, useful for stabilizing pharmaceuticals or analytical proteins, comprises a filler, stabilizer, buffer and optionally nonionic surfactant |
US7956160B2 (en) * | 2005-07-22 | 2011-06-07 | Amgen Inc. | Concentrated protein lyophilates, methods, and uses |
JP5555425B2 (en) | 2005-11-01 | 2014-07-23 | ワイス・エルエルシー | Sodium chloride solution for drug reconstitution or dilution |
EP2299271B9 (en) * | 2005-11-09 | 2014-02-26 | Ajinomoto Co., Inc. | Kokumi-imparting compositions |
US8420144B2 (en) * | 2005-11-09 | 2013-04-16 | Ajinomoto Co., Inc. | Kokumi-imparting agent, method of using, and compositions containing same |
WO2007055388A2 (en) * | 2005-11-09 | 2007-05-18 | Ajinomoto Co., Inc. | Calcium receptor activator |
RU2008118166A (en) * | 2005-11-22 | 2009-12-27 | Вайет (Us) | COMPOSITIONS CONTAINING HYBRID PROTEINS INCLUDING IMMUNOGLOBULIN |
PT1969004E (en) | 2005-12-28 | 2011-11-25 | Novo Nordisk As | Compositions comprising an acylated insulin and zinc and method of making the said compositions |
EP2010222A1 (en) | 2006-03-31 | 2009-01-07 | Baxter International Inc. | Pegylated factor viii |
US7985839B2 (en) | 2006-03-31 | 2011-07-26 | Baxter International Inc. | Factor VIII polymer conjugates |
US7982010B2 (en) | 2006-03-31 | 2011-07-19 | Baxter International Inc. | Factor VIII polymer conjugates |
US7645860B2 (en) | 2006-03-31 | 2010-01-12 | Baxter Healthcare S.A. | Factor VIII polymer conjugates |
JP5094850B2 (en) * | 2006-05-31 | 2012-12-12 | ジェンザイム コーポレーション | Use of polysaccharides to promote enzyme activity |
CN103933612B (en) | 2006-10-27 | 2016-06-22 | 爱德华兹生命科学公司 | Biological tissue for Srgery grafting |
AR063606A1 (en) * | 2006-11-07 | 2009-02-04 | Acambis Inc | VACCINE STABILIZATION THROUGH LIOFILIZATION |
JP5876208B2 (en) | 2006-12-15 | 2016-03-02 | バクスター・インターナショナル・インコーポレイテッドBaxter International Incorp0Rated | Factor VIIa- (poly) sialic acid conjugate with extended in vivo half-life |
FR2913020B1 (en) | 2007-02-23 | 2012-11-23 | Biomethodes | NEW VIII FACTORS FOR THE TREATMENT OF TYPE A HEMOPHILS |
WO2008107908A1 (en) * | 2007-03-05 | 2008-09-12 | Cadila Healthcare Limited | Compositions comprising peg- interferon alpha conjugates and raffinose as cryoprotectant |
AU2013204652B2 (en) * | 2007-04-26 | 2015-06-18 | Bayer Healthcare Llc | Stabilization of liquid solutions of recombinant protein for frozen storage |
RU2469739C2 (en) * | 2007-04-26 | 2012-12-20 | БАЙЕР ХЕЛСКЕА ЛЛСи | Stabilisation of liquid solutions of recombinant protein for storing in frozen state |
EP2156753A4 (en) * | 2007-05-08 | 2010-11-24 | Ajinomoto Kk | Low-fat food |
US9101691B2 (en) | 2007-06-11 | 2015-08-11 | Edwards Lifesciences Corporation | Methods for pre-stressing and capping bioprosthetic tissue |
US9034818B2 (en) * | 2007-06-13 | 2015-05-19 | Novo Nordisk A/S | Pharmaceutical formulations comprising an insulin derivative |
GB0715285D0 (en) * | 2007-08-06 | 2007-09-12 | Britannia Pharmaceuticals Ltd | Improvements in or relating to powdered medicaments for nasal delivery |
CN101376022B (en) * | 2007-08-31 | 2011-11-30 | 上海医药工业研究院 | Medicament composition containing defibrase modified by PEG |
CA2991162A1 (en) * | 2007-12-21 | 2009-07-02 | Aptevo Biotherapeutics Llc | Stabilized factor ix formulations containing trehalose |
US8357387B2 (en) | 2007-12-21 | 2013-01-22 | Edwards Lifesciences Corporation | Capping bioprosthetic tissue to reduce calcification |
SG10201503304RA (en) * | 2007-12-27 | 2015-06-29 | Baxter Int | Cell culture processes |
EP2113564A1 (en) * | 2008-05-01 | 2009-11-04 | Arecor Limited | Protein formulation |
CA2729972C (en) * | 2008-08-05 | 2018-11-20 | Wyeth Llc | Lyophilization above collapse |
AU2009284113B2 (en) | 2008-08-21 | 2015-05-14 | Octapharma Ag | Recombinantly produced human factor VIII and IX |
MX2011002159A (en) * | 2008-08-27 | 2011-03-29 | Schering Corp | Lyophilized formulatons of engineered anti-il-23p19 antibodies. |
EP2337580B1 (en) * | 2008-09-03 | 2012-03-28 | Octapharma AG | Stabilized compositions for recombinantly produced factor viii |
US20100068210A1 (en) * | 2008-09-10 | 2010-03-18 | Ji Junyan A | Compositions and methods for the prevention of oxidative degradation of proteins |
AU2009309623B9 (en) * | 2008-10-30 | 2014-10-02 | Novo Nordisk A/S | Treating diabetes melitus using insulin injections with less than daily injection frequency |
BRPI0921429B1 (en) | 2008-11-07 | 2022-07-12 | Takeda Pharmaceutical Company Limited | STABLE LYOPHILIZED PHARMACEUTICAL FORMULATION, AND METHOD FOR PREPARING A STABLE LYOPHILIZED FACTOR |
EP2248518B1 (en) | 2009-04-17 | 2013-01-16 | Merz Pharma GmbH & Co. KGaA | Formulation for stabilizing proteins, peptides or mixtures thereof. |
US8809501B2 (en) | 2009-07-27 | 2014-08-19 | Baxter International Inc. | Nucleophilic catalysts for oxime linkage |
EP3081233B1 (en) | 2009-07-27 | 2020-12-23 | Baxalta GmbH | Glycopolysialylation of proteins other than blood coagulation proteins |
US8642737B2 (en) | 2010-07-26 | 2014-02-04 | Baxter International Inc. | Nucleophilic catalysts for oxime linkage |
DK2459224T3 (en) | 2009-07-27 | 2016-07-25 | Baxalta GmbH | Blodstørkningsproteinkonjugater |
ES2856055T3 (en) | 2009-07-27 | 2021-09-27 | Baxalta GmbH | Glycopolysialylation of proteins other than blood clotting proteins |
GB0915480D0 (en) | 2009-09-04 | 2009-10-07 | Arecor Ltd | Stable formulation of factor viii |
ES2865250T3 (en) | 2009-09-21 | 2021-10-15 | Takeda Pharmaceuticals Co | Stabilized Liquid and Freeze-Dried ADAMTS13 Formulations |
EP2496246B1 (en) | 2009-11-03 | 2018-06-27 | Grifols Therapeutics LLC | Composition, method, and kit for alpha-1 proteinase inhibitor |
HUE037733T2 (en) * | 2009-11-24 | 2018-09-28 | Grifols Therapeutics Inc | Lyophilization methods, compositions, and kits |
RU2012135505A (en) * | 2010-01-19 | 2014-02-27 | Ханми Сайенс Ко., Лтд. | LIQUID COMPOSITIONS FOR LONG-TERM ETHYTROPOETIN CONJUGATE |
EP2549957B1 (en) | 2010-03-23 | 2019-01-30 | Edwards Lifesciences Corporation | Methods of conditioning sheet bioprosthetic tissue |
US8906601B2 (en) | 2010-06-17 | 2014-12-09 | Edwardss Lifesciences Corporation | Methods for stabilizing a bioprosthetic tissue by chemical modification of antigenic carbohydrates |
TW201213342A (en) * | 2010-09-17 | 2012-04-01 | Baxter Int | Stabilization of immunoglobulins and other proteins through aqueous formulation with sodium chloride at weak acidic to neutral pH |
US9173944B2 (en) | 2010-10-12 | 2015-11-03 | Merz Pharma Gmbh & Co. Kgaa | Formulation suitable for stabilizing proteins, which is free of mammalian excipients |
JP6049625B2 (en) | 2010-10-27 | 2016-12-21 | ノヴォ ノルディスク アー/エス | Treatment of diabetes mellitus using insulin injections given at various injection intervals |
EA035447B1 (en) | 2010-11-05 | 2020-06-17 | Баксалта Инкорпорейтид | Isolated recombinant partially b-domain deleted factor viii (fviii) variant |
US9351829B2 (en) | 2010-11-17 | 2016-05-31 | Edwards Lifesciences Corporation | Double cross-linkage process to enhance post-implantation bioprosthetic tissue durability |
WO2012079979A1 (en) * | 2010-12-16 | 2012-06-21 | Novo Nordisk A/S | Aqueous factor viii solution |
PL2654794T3 (en) | 2010-12-22 | 2020-07-27 | Baxalta GmbH | Materials and methods for conjugating a water soluble fatty acid derivative to a protein |
BRPI1105317A2 (en) | 2011-01-24 | 2013-04-30 | Fundacco Hemoct De Ribeirco Preto | stable and large-scale production of human fviii in human cell line sk-hep-1 |
KR102289394B1 (en) | 2011-03-31 | 2021-08-13 | 머크 샤프 앤드 돔 코포레이션 | Stable formulations of antibodies to human programmed death receptor pd-1 and related treatments |
JP6060447B2 (en) | 2011-04-08 | 2017-01-18 | バイオ−ラッド ラボラトリーズ インコーポレーティッド | Sso7 polymerase conjugate with reduced non-specific activity |
CN103703141B (en) | 2011-04-08 | 2016-08-17 | 伯乐实验室公司 | There is the PCR reactant mixture of the nonspecific activity of reduction |
US20140227250A1 (en) | 2012-08-23 | 2014-08-14 | Merck Sharp & Dohme Corp. | Stable formulations of antibodies to tslp |
US10238771B2 (en) | 2012-11-08 | 2019-03-26 | Edwards Lifesciences Corporation | Methods for treating bioprosthetic tissue using a nucleophile/electrophile in a catalytic system |
CN105209487A (en) * | 2013-03-15 | 2015-12-30 | 拜耳医药保健有限公司 | Recombinant Factor VIII formulations |
CA2899737A1 (en) * | 2013-03-15 | 2014-09-18 | Biogen Ma Inc. | Factor viii polypeptide formulations |
US9839579B2 (en) | 2013-04-24 | 2017-12-12 | Corning Incorporated | Delamination resistant pharmaceutical glass containers containing active pharmaceutical ingredients |
US9700486B2 (en) | 2013-04-24 | 2017-07-11 | Corning Incorporated | Delamination resistant pharmaceutical glass containers containing active pharmaceutical ingredients |
EP2991672A1 (en) | 2013-04-30 | 2016-03-09 | Novo Nordisk A/S | Novel administration regime |
KR102058864B1 (en) * | 2013-12-16 | 2019-12-24 | 주식회사 티움바이오 | Pharmaceutical composition having improved stability comprising a fusion protein of factor vii |
MX2016012870A (en) * | 2014-04-01 | 2017-05-12 | Advantech Bioscience Farmacêutica Ltda | Stable factor viii formulations with low sugar-glycine. |
US9855319B2 (en) | 2014-04-01 | 2018-01-02 | Advantech Bioscience Farmacêutica Ltda | Stabilization of factor VIII without calcium as an excipient |
AU2015299224B2 (en) * | 2014-08-04 | 2019-06-20 | Csl Limited | Factor VIII formulation |
CA2961739A1 (en) * | 2014-08-20 | 2016-02-25 | Portola Pharmaceuticals, Inc. | Lyophilized formulations for factor xa antidote |
CN114129709A (en) * | 2014-10-23 | 2022-03-04 | 恩格姆生物制药公司 | Pharmaceutical compositions comprising peptide variants and methods of use thereof |
DK3287140T3 (en) | 2015-04-21 | 2021-07-19 | Beijing Staidson Medical Tech Co Ltd | Nerve growth factor composition and powder injection |
US10576129B2 (en) | 2015-04-21 | 2020-03-03 | Staidson (Beijing) Biopharmaceuticals Co., Ltd. | Nerve growth factor composition and powder injection |
US20170007694A1 (en) * | 2015-07-07 | 2017-01-12 | Nanobio Corporation | Methods and compositions for the stabilization of proteins |
GB2556002B (en) | 2015-07-07 | 2020-12-16 | Bluewillow Biologics Inc | Methods and compositions for nanoemulsion vaccine formulations |
WO2017011275A1 (en) * | 2015-07-10 | 2017-01-19 | Nersissian Aram M | Factor viii protein compositions and methods of treating hemophilia a |
EP3167877A1 (en) * | 2015-11-12 | 2017-05-17 | Bayer Pharma Aktiengesellschaft | Method for the production of freeze-dried pellets comprising factor viii |
US20190046450A1 (en) * | 2016-02-24 | 2019-02-14 | Portola Pharmaceuticals, Inc. | Lyophilized formulations for factor xa antidote |
CN109640961B (en) * | 2016-06-01 | 2021-11-02 | 施维雅知识产权英国有限公司 | Polyalkylene oxide-asparaginase formulations and methods of making and using same |
CN110381962B (en) * | 2016-12-21 | 2023-10-13 | 孙崇谨 | Novel method for the active preservation of plasma proteins |
JOP20190260A1 (en) | 2017-05-02 | 2019-10-31 | Merck Sharp & Dohme | Stable formulations of programmed death receptor 1 (pd-1) antibodies and methods of use thereof |
SG11201909955XA (en) | 2017-05-02 | 2019-11-28 | Merck Sharp & Dohme | Formulations of anti-lag3 antibodies and co-formulations of anti-lag3 antibodies and anti-pd-1 antibodies |
JP6630315B2 (en) | 2017-06-27 | 2020-01-15 | 矢崎総業株式会社 | Noise reduction unit |
US10335464B1 (en) | 2018-06-26 | 2019-07-02 | Novo Nordisk A/S | Device for titrating basal insulin |
AU2020204922A1 (en) * | 2019-01-06 | 2021-07-01 | Endo Global Aesthetics Limited | Collagenase formulations and methods of producing the same |
EP3994162A1 (en) | 2019-07-04 | 2022-05-11 | CSL Behring Lengnau AG | A truncated von willebrand factor (vwf) for increasing the in vitro stability of coagulation factor viii |
CN110772487B (en) * | 2019-12-09 | 2021-09-21 | 湖南科伦制药有限公司 | Freeze-drying method of ethylenediamine diaceturate |
CN112121009B (en) * | 2020-09-24 | 2022-12-02 | 科兴生物制药股份有限公司 | New preparation of polyethylene glycol modified recombinant human granulocyte stimulating factor |
EP4217394A1 (en) | 2020-09-24 | 2023-08-02 | Merck Sharp & Dohme LLC | Stable formulations of programmed death receptor 1 (pd-1) antibodies and hyaluronidase variants and fragments thereof and methods of use thereof |
EP4240757A2 (en) | 2020-11-09 | 2023-09-13 | Takeda Pharmaceutical Company Limited | Purification of fviii from plasma using silicon oxide adsorption |
WO2022217041A1 (en) * | 2021-04-09 | 2022-10-13 | Hyalo Technologies, LLC | Method of sterilization of biologics |
AU2022326188A1 (en) * | 2021-08-11 | 2024-02-22 | Grifols Worldwide Operations Limited | Method for producing human plasma-derived factor viii / von willebrand factor and composition obtained |
CN114015758B (en) * | 2021-10-15 | 2022-06-24 | 无锡百泰克生物技术有限公司 | Freeze-drying protective agent, fluorescent PCR detection kit and freeze-drying process |
Family Cites Families (177)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB941019A (en) | 1961-04-27 | 1963-11-06 | Crookes Lab Ltd | Improvements in and relating to the stability of anti-haemophilic globulin (factor viii) |
US3389314A (en) | 1962-05-07 | 1968-06-18 | Penn Controls | Proportional silicon controlled rectifier driven system for a heat motor |
US3681126A (en) | 1969-11-25 | 1972-08-01 | Monsanto Co | Flame retardant article containing tris-(3 - halo - 2-hydroxypropyl)-hydroxymethylphosphonium chloride |
US3839314A (en) | 1971-06-29 | 1974-10-01 | Baxter Laboratories Inc | Clarification of blood serum and plasma using block copolymers of ethylene oxide and polyoxypropylene |
US3770631A (en) | 1971-06-29 | 1973-11-06 | Baxter Laboratories Inc | Clarification of blood serum and plasma |
US4073886A (en) | 1973-01-30 | 1978-02-14 | Baxter Travenol Laboratories, Inc. | Blood fractionation process using block copolymers of ethylene oxide and polyoxypropylene |
US3980432A (en) | 1973-04-02 | 1976-09-14 | Behringwerke Aktiengesellschaft | Partial thromboplastin, its use as diagnostic agent and process for preparing it |
US3893990A (en) | 1973-04-05 | 1975-07-08 | Baxter Laboratories Inc | Clarification of blood serum and plasma using a block copolymer of ethylene oxide and polyoxypropylene |
US3893991A (en) | 1973-04-05 | 1975-07-08 | Baxter Laboratories Inc | Clarification of blood serum and plasma using block copolymers of ethylene oxide and polyoxypropylene |
US4189425A (en) | 1975-04-11 | 1980-02-19 | Edward Shanbrom, Inc. | Method of preserving blood plasma I |
US4086218A (en) | 1975-04-11 | 1978-04-25 | Edward Shanbrom, Inc. | Method of preserving blood plasma II |
US4069216A (en) | 1975-06-16 | 1978-01-17 | Edward Shanbrom, Inc. | Simplified methods for preparation of very high purity Factor VIII concentrate |
US4089944A (en) | 1975-12-22 | 1978-05-16 | Baxter Travenol Laboratories, Inc. | Rapidly solubilized AHF composition and process for preparing same |
US4027013A (en) | 1976-01-22 | 1977-05-31 | William L. Wilson | Clottable fibrinogen free factor VIII and albumin product and process |
SU663404A1 (en) | 1976-12-30 | 1979-05-25 | Московский Ордена Ленина И Ордена Трудового Красного Знамени Государственный Университет Им. М.В.Ломоносова | Method of obtaining fibrin monomer |
US4137223A (en) | 1977-05-16 | 1979-01-30 | Edward Shanbrom, Inc. | Method of preserving blood plasma II |
SE443293B (en) | 1978-01-25 | 1986-02-24 | Blombaeck E G B | Blood fraction FRONT TELL NING |
DE2916711A1 (en) | 1979-04-25 | 1980-11-06 | Behringwerke Ag | Blood coagulation factors and process for their manufacture |
US4783441A (en) | 1979-04-30 | 1988-11-08 | Hoechst Aktiengesellschaft | Aqueous protein solutions stable to denaturation |
FR2460305A2 (en) | 1979-06-29 | 1981-01-23 | Merieux Inst | PROCESS FOR PREPARING A FACTOR VIII CONCENTRATE |
US4386068A (en) | 1980-02-26 | 1983-05-31 | Cutter Laboratories, Inc. | Antihemophilic factor concentrate and method for preparation |
US4623717A (en) | 1980-03-05 | 1986-11-18 | Miles Laboratories, Inc. | Pasteurized therapeutically active protein compositions |
US4341764A (en) | 1980-03-05 | 1982-07-27 | Cutter Laboratories, Inc. | Method of preparing fibronectin and antihemophilic factor |
DE3176491D1 (en) | 1980-03-05 | 1987-11-26 | Miles Lab | Pasteurized therapeutically active protein compositions |
US4440679A (en) | 1980-03-05 | 1984-04-03 | Cutter Laboratories, Inc. | Pasteurized therapeutically active protein compositions |
JPS56127308A (en) | 1980-03-11 | 1981-10-06 | Green Cross Corp:The | Blood-coagulation factor 8 pharmaceutical |
JPS56135418A (en) | 1980-03-27 | 1981-10-22 | Green Cross Corp:The | Heat treatment of aqueous solution containing 8 factor of coagulation of blood derived from human |
AT369263B (en) | 1980-08-27 | 1982-12-27 | Immuno Ag | METHOD FOR PRODUCING A FACTOR VIII (AHF) HIGH CONCENTRATE |
DE3033932C2 (en) | 1980-09-10 | 1984-05-24 | Biotest-Serum-Institut Gmbh, 6000 Frankfurt | Process for the cold sterilization of preparations containing blood coagulation factor VIII |
JPS5770814A (en) | 1980-10-17 | 1982-05-01 | Isamu Horikoshi | Oral preparation of blood clotting eighth factor |
US4495278A (en) | 1981-04-27 | 1985-01-22 | Baxter Travenol Laboratories, Inc. | Process for making novel blood clotting enzyme compositions |
US4382083A (en) | 1981-06-25 | 1983-05-03 | Baxter Travenol Laboratories, Inc. | Therapeutic method for treating blood-clotting defects with factor VIIa |
US4479938A (en) | 1981-06-25 | 1984-10-30 | Baxter Travenol Laboratories, Inc. | Therapeutic composition containing factor VIIa |
JPS5874617A (en) | 1981-10-28 | 1983-05-06 | Green Cross Corp:The | Heat-treating method of aqueous solution containing blood coagulation factor 8 derived from human |
US4456590B2 (en) | 1981-11-02 | 1989-05-30 | Heat treatment of lyphilized blood clotting factor viii concentrate | |
US4481189A (en) | 1982-04-14 | 1984-11-06 | New York Blood Center Inc. | Process for preparing sterilized plasma and plasma derivatives |
US4591505A (en) | 1982-04-14 | 1986-05-27 | New York Blood Center, Inc. | Process for inactivating hepatitis B virus |
US4495175A (en) | 1982-08-05 | 1985-01-22 | University Of Rochester | Preparation of highly purified human antihemophilic factor |
DE3230849A1 (en) | 1982-08-19 | 1984-02-23 | Behringwerke Ag, 3550 Marburg | PASTEURIZED HUMAN FIBRINOGEN (HF) AND METHOD FOR THE PRODUCTION THEREOF |
DE3237512A1 (en) | 1982-10-09 | 1984-04-12 | Behringwerke Ag, 3550 Marburg | METHOD FOR PASTEURIZING ANTIHAEMOPHILIC CRYOPRAEZIPITATE (AHK) AND ANTIHAEMOPHILE CRYOPRAECIPITATE PRODUCED THEREOF |
GB2129685B (en) | 1982-11-11 | 1985-11-13 | Nat Biolog Standards Board | Anti-haemophilic compositions |
JPS59134730A (en) | 1983-01-20 | 1984-08-02 | Green Cross Corp:The | Heat treatment of viii factor of blood clotting |
ZA842418B (en) | 1983-03-31 | 1984-11-28 | Scripps Clinic Res | Factor viii coagulant polypeptides and monoclonal antibodies to them |
US4748120A (en) | 1983-05-02 | 1988-05-31 | Diamond Scientific Co. | Photochemical decontamination treatment of whole blood or blood components |
US4727027A (en) | 1983-05-02 | 1988-02-23 | Diamond Scientific Co. | Photochemical decontamination treatment of whole blood or blood components |
DE3481109D1 (en) | 1983-05-09 | 1990-03-01 | Novo Nordisk As | ANTIHAEMOPHILIA FACTOR VIII CONCENTRATE AND THEIR PRODUCTION METHOD. |
AT379510B (en) | 1983-05-20 | 1986-01-27 | Immuno Ag | METHOD FOR PRODUCING A FACTOR VIII (AHF) CONTAINING PRAEPARATION |
DE3318521A1 (en) | 1983-05-20 | 1984-11-22 | Lentia GmbH Chem. u. pharm. Erzeugnisse - Industriebedarf, 8000 München | METHOD FOR PRODUCING AN ANTIHAEMOPHILIE FACTOR CONCENTRATE |
US4540573A (en) | 1983-07-14 | 1985-09-10 | New York Blood Center, Inc. | Undenatured virus-free biologically active protein derivatives |
DE3336631A1 (en) | 1983-10-08 | 1985-04-18 | Behringwerke Ag, 3550 Marburg | METHOD FOR THE PASTEURIZATION OF PLASMA OR CONCENTRATES OF THE BLOOD COAGINING FACTORS II, VII, IX AND X |
US4757006A (en) | 1983-10-28 | 1988-07-12 | Genetics Institute, Inc. | Human factor VIII:C gene and recombinant methods for production |
US4693981A (en) | 1983-12-20 | 1987-09-15 | Advanced Genetics Research Institute | Preparation of inactivated viral vaccines |
JPS6122022A (en) | 1983-12-28 | 1986-01-30 | Green Cross Corp:The | Method for heat-treating blood plasma protein |
DE10399009I1 (en) | 1984-01-12 | 2003-10-23 | Chiron Corp | DNA sequences encoding factor VIIIc and related DNA constructions |
AT389815B (en) | 1984-03-09 | 1990-02-12 | Immuno Ag | METHOD FOR INACTIVATING VARIABLE FILTERABLE DISEASERS IN BLOOD PRODUCTS |
US4831012A (en) | 1984-03-23 | 1989-05-16 | Baxter International Inc. | Purified hemoglobin solutions and method for making same |
JPS60199829A (en) | 1984-03-24 | 1985-10-09 | Nippon Sekijiyuujishiya | Preparation of high-purity antihemophilic globulin |
US5043428A (en) | 1984-08-31 | 1991-08-27 | Behringwerke Aktiengesellschaft | Pasteurized, isoagglutinin-free factor VIII preparation and a process for its production |
US4543210A (en) | 1984-10-04 | 1985-09-24 | Miles Laboratories, Inc. | Process for producing a high purity antihemophilic factor concentrate |
US4613501A (en) | 1984-12-21 | 1986-09-23 | New York Blood Center, Inc. | Inactivation of viruses in labile blood derivatives |
JPS61271222A (en) * | 1985-05-24 | 1986-12-01 | Dainippon Pharmaceut Co Ltd | Human interleukine 1 polypeptide and derivative thereof |
US4952675A (en) | 1985-02-01 | 1990-08-28 | New York University | Method for purifying antihemophilic factor |
US4847362A (en) | 1985-02-01 | 1989-07-11 | New York University | Method for purifying antihemophilic factor |
US4743680A (en) | 1985-02-01 | 1988-05-10 | New York University | Method for purifying antihemophilic factor |
EP0218712B1 (en) | 1985-04-12 | 1992-02-26 | Genetics Institute, Inc. | Novel procoagulant proteins |
US4891319A (en) | 1985-07-09 | 1990-01-02 | Quadrant Bioresources Limited | Protection of proteins and the like |
US4758657A (en) | 1985-07-11 | 1988-07-19 | Armour Pharmaceutical Company | Method of purifying Factor VIII:C |
EP0212040B1 (en) | 1985-08-05 | 1990-05-23 | IMMUNO Aktiengesellschaft für chemisch-medizinische Produkte | Process for producing preparations based on blood coagulation factors |
AU6487286A (en) | 1985-11-08 | 1987-05-14 | Baxter Travenol Laboratories Inc. | Antihemophilic factor by cold precipitation |
US5180583A (en) | 1985-11-26 | 1993-01-19 | Hedner Ulla K E | Method for the treatment of bleeding disorders |
JPS62195331A (en) | 1986-02-24 | 1987-08-28 | Nippon Sekijiyuujishiya | Production of blood coagulation factor viii pharmaceutical |
AT390374B (en) | 1986-03-18 | 1990-04-25 | Schwab & Co Gmbh | METHOD FOR PASTEURIZING PLASMA PROTEIN AND PLASMA PROTEIN FRACTIONS |
DE3609431A1 (en) | 1986-03-20 | 1987-09-24 | Biotest Pharma Gmbh | METHOD FOR PRODUCING A STERILE PRESERVATIVE CONTAINING THE BLOOD coagulation factor VIII |
US4841023A (en) | 1986-06-25 | 1989-06-20 | New York Blood Center, Inc. | Inactivation of viruses in labile protein-containing compositions using fatty acids |
AT391808B (en) | 1986-11-03 | 1990-12-10 | Immuno Ag | METHOD FOR PRODUCING A FACTOR VIII (AHF) CONTAINING FRACTION |
GB8628104D0 (en) | 1986-11-25 | 1986-12-31 | Connaught Lab | Pasteurization of immunoglobin solutions |
CA1331157C (en) | 1987-04-06 | 1994-08-02 | Randal J. Kaufman | Method for producing factor viii:c-type proteins |
ES2006632A6 (en) | 1987-04-21 | 1989-05-01 | Green Cross Corp | Process for heat treating fibrinogen |
US4876241A (en) | 1987-05-22 | 1989-10-24 | Armour Pharmaceutical Company | Stabilization of biological and pharmaceutical products during thermal inactivation of viral and bacterial contaminants |
IL86417A (en) | 1987-05-22 | 1992-09-06 | Armour Pharma | Process for the inactivation of pathogens in biological or pharmaceutical material by mixing with aqueous solution containing a sugar(alcohol)and neutral salts as stabilizers |
US4795806A (en) | 1987-07-16 | 1989-01-03 | Miles Laboratories, Inc. | Phospholipid affinity purification of Factor VIII:C |
JPH0387173A (en) | 1987-09-10 | 1991-04-11 | Teijin Ltd | Preparation of human active natural type factor viii c and transformant using the same |
DE3730533A1 (en) | 1987-09-11 | 1989-03-30 | Biotest Pharma Gmbh | METHOD FOR STERILIZING PLASMA OR PLASMA FACTIONS |
KR890004724A (en) * | 1987-09-17 | 1989-05-09 | 히사시 미하라 | New protease preparations |
CA1329760C (en) | 1987-10-29 | 1994-05-24 | Ted C. K. Lee | Plasma and recombinant protein formulations in high ionic strength media |
US5605884A (en) | 1987-10-29 | 1997-02-25 | Rhone-Poulenc Rorer Pharmaceuticals Inc. | Factor VIII formulations in high ionic strength media |
US4877608A (en) | 1987-11-09 | 1989-10-31 | Rorer Pharmaceutical Corporation | Pharmaceutical plasma protein formulations in low ionic strength media |
US5177191A (en) | 1987-12-21 | 1993-01-05 | Miles, Inc. | Gel filtration of factor VIII |
DK18288D0 (en) | 1988-01-15 | 1988-01-15 | Nordisk Gentofte | PROCEDURE FOR PASTEURIZATION OF Aqueous SOLUTIONS OF FACTOR VIII |
WO1989009784A1 (en) | 1988-04-08 | 1989-10-19 | Commonwealth Serum Laboratories Commission | Production of heat-stable factor viii concentrate |
US4981951A (en) | 1988-04-14 | 1991-01-01 | Miles Inc. | Lectin affinity chromatography of factor VIII |
FR2632309B1 (en) | 1988-06-07 | 1990-08-24 | Lille Transfusion Sanguine | PROCESS FOR THE CHROMATOGRAPHIC PURIFICATION OF PROTEINS, PARTICULARLY FACTOR VIII, AND THE PRODUCTS OBTAINED |
US5047249A (en) | 1988-07-22 | 1991-09-10 | John Morris Co., Inc. | Compositions and methods for treating skin conditions and promoting wound healing |
US5051353A (en) | 1988-08-09 | 1991-09-24 | The United States Of America As Represented By The Secretary Of The Navy | Preservation and restoration of hemoglobin in blood substitutes |
JPH02157231A (en) * | 1988-12-07 | 1990-06-18 | Bio Kagaku Kenkyusho:Kk | Cytostatic agent |
DE3904354A1 (en) | 1989-02-14 | 1990-08-16 | Behringwerke Ag | PASTEURIZED, PURIFIED OF WILLEBRAND FACTOR CONCENTRATE AND METHOD FOR THE PRODUCTION THEREOF |
FR2665449B1 (en) | 1990-08-02 | 1995-04-14 | Aquitaine Developp Transf Sang | METHOD FOR MANUFACTURING A VON WILLEBRAND FACTOR HAVING VERY HIGH PURITY, MAINLY LACKING AN ANTIHEMOPHILIC FACTOR (FVIIIC), AND VON WILLEBRAND FACTOR THUS OBTAINED, AS WELL AS A PHARMACEUTICAL COMPOSITION CONTAINING THE SAME. |
US5760183A (en) | 1989-02-17 | 1998-06-02 | Association D'aquitaine Pour De Developpment De La Transfusion Sanguine Et Des Recherches Hematologiques | Process for the manufacture of very high-purity antithaemophilic factor (FVIIIC), and von Willebrand factor, and pharmaceutical compositions containing same |
CN1047342A (en) | 1989-05-13 | 1990-11-28 | 杭州市中心血站 | The production of Factor IX and virus inactivating method thereof |
DK0399321T3 (en) | 1989-05-24 | 1993-08-09 | Miles Inc | Gel filtration of heat treated factor VIII |
GB8913183D0 (en) * | 1989-06-08 | 1989-07-26 | Central Blood Lab Authority | Chemical products |
US5138034A (en) | 1989-07-12 | 1992-08-11 | The Green Cross Corporation | Method of fractionating plasma proteins |
US5062498A (en) | 1989-07-18 | 1991-11-05 | Jaromir Tobias | Hydrostatic power transfer system with isolating accumulator |
DK0410207T3 (en) | 1989-07-24 | 1997-07-14 | Bayer Ag | Stabilization of Highly Purified Proteins. |
DE3926034C3 (en) | 1989-08-07 | 1996-11-21 | Behringwerke Ag | Process for the preparation of a stable factor VIII |
FR2651437A1 (en) | 1989-09-05 | 1991-03-08 | Lille Transfusion Sanguine | PROCESS FOR PREPARING A CONCENTRATE OF THE VON WILLEBRAND FACTOR VIII-FACTOR COMPLEX OF BLOOD COAGULATION FROM TOTAL PLASMA. |
DK162233C (en) | 1989-11-09 | 1992-03-16 | Novo Nordisk As | PROCEDURE FOR INSULATING FACTOR VIII FROM BLOOD PLASMA AND PHARMACEUTICAL PREPARATION CONTAINING THE ASSOCIATED PHATAR VIII |
SE465222C5 (en) | 1989-12-15 | 1998-02-10 | Pharmacia & Upjohn Ab | A recombinant human factor VIII derivative and process for its preparation |
US5439882A (en) | 1989-12-29 | 1995-08-08 | Texas Tech University Health Sciences Center | Blood substitute |
DE4001451A1 (en) | 1990-01-19 | 1991-08-01 | Octapharma Ag | STABLE INJECTABLE SOLUTIONS OF FACTOR VIII AND FACTOR IX |
US5587490A (en) | 1990-04-16 | 1996-12-24 | Credit Managers Association Of California | Method of inactivation of viral and bacterial blood contaminants |
US5418130A (en) | 1990-04-16 | 1995-05-23 | Cryopharm Corporation | Method of inactivation of viral and bacterial blood contaminants |
US5798238A (en) | 1990-04-16 | 1998-08-25 | Baxter International Inc. | Method of inactivation of viral and bacterial blood contaminants with quinolines as photosensitizer |
WO1991017194A1 (en) * | 1990-05-07 | 1991-11-14 | Exxon Chemical Patents Inc. | UNSATURATED α-OLEFIN COPOLYMERS AND METHOD FOR PREPARATION THEREOF |
US5378612A (en) | 1990-05-11 | 1995-01-03 | Juridical Foundation The Chemo-Sero-Therapeutic Research Institute | Culture medium for production of recombinant protein |
US5712086A (en) | 1990-05-15 | 1998-01-27 | New York Blood Center, Inc. | Process for transfusing cell containing fractions sterilized with radiation and a quencher of type I and type II photodynamic reactions |
US5232844A (en) | 1990-05-15 | 1993-08-03 | New York Blood Center | Photodynamic inactivation of viruses in cell-containing compositions |
FR2662166A1 (en) | 1990-05-18 | 1991-11-22 | Fondation Nale Transfusion San | PROCESS FOR PREPARING HIGHLY HIGH PURITY FACTOR VIII COMPRISING A RAPID IMMUNOADSORPTION STEP |
IT1248723B (en) | 1990-06-12 | 1995-01-26 | Scalvo S P A | PROCESS FOR THE PURIFICATION OF FACTOR VIII AND FACTOR VIII OBTAINED WITH SUCH PROCESS |
JP3127263B2 (en) | 1990-07-12 | 2001-01-22 | バクスター、ダイアグノスティックス、インコーポレイテッド | Part VIII: Ca factor chromogen assay |
AU650045B2 (en) | 1990-09-12 | 1994-06-09 | Lifecell Corporation | Method and apparatus for cryopreparation dry stabilization and rehydration of biological suspensions |
US5272135A (en) | 1991-03-01 | 1993-12-21 | Chiron Ophthalmics, Inc. | Method for the stabilization of methionine-containing polypeptides |
FR2673632A1 (en) | 1991-03-08 | 1992-09-11 | Lille Transfusion Sanguine | PROCESS FOR THE PREPARATION OF HUMAN VON WILLEBRAND FACTOR CONCENTRATE OF HIGH PURITY, SUITABLE FOR THERAPEUTIC USE |
DE4111393A1 (en) | 1991-04-09 | 1992-10-15 | Behringwerke Ag | STABILIZED FACTOR VIII PREPARATIONS |
SE468480B (en) | 1991-05-24 | 1993-01-25 | Arne Holmgren | Modified thioredoxin and its use |
AU2309692A (en) | 1991-07-03 | 1993-02-11 | Cryolife, Inc. | Method for stabilization of biomaterials |
US5254350A (en) | 1991-07-22 | 1993-10-19 | Helena Laboratories Corporation | Method of preparing a thromboplastin extract |
CA2078721A1 (en) | 1991-09-24 | 1993-03-25 | Hiroshi Yonemura | Process for preparing human coagulation factor viii protein complex |
FR2681867B1 (en) | 1991-09-26 | 1993-12-31 | Pasteur Merieux Serums Vaccins | FACTOR VIII PURIFICATION PROCESS AND PREPARATIONS OBTAINED. |
US5278289A (en) | 1991-11-12 | 1994-01-11 | Johnson Alan J | Antihemophilic factor stabilization |
US5192743A (en) | 1992-01-16 | 1993-03-09 | Genentech, Inc. | Reconstitutable lyophilized protein formulation |
JPH05331071A (en) * | 1992-01-17 | 1993-12-14 | Asahi Chem Ind Co Ltd | Lyophilized composition of calcitonin gene-related peptide and stabilization thereof |
AT399818B (en) | 1992-04-24 | 1995-07-25 | Immuno Ag | METHOD FOR PRODUCING A HIGH PURIFIED VIRUS-SAFE FACTOR VIII PREPARATION |
EP0638091B1 (en) | 1992-04-30 | 2005-12-07 | Probitas Pharma Inc. | Improved solubilization and stabilization of factor viii complex |
US5288853A (en) | 1992-04-30 | 1994-02-22 | Alpha Therapeutic Corporation | Factor viii purification process |
US5378601A (en) | 1992-07-24 | 1995-01-03 | Montefiore Medical Center | Method of preserving platelets with apyrase and an antioxidant |
US5424471A (en) * | 1992-07-31 | 1995-06-13 | U.S. Bioscience, Inc. | Crystalline amifostine compositions and methods of the preparation and use of same |
EP1652534A3 (en) * | 1992-10-02 | 2007-05-02 | Genetics Institute, LLC | Composition comprising coagulation factor VIII formulation, process for its preparation and use of a surfactant as stabilizer |
IT1256622B (en) | 1992-12-04 | 1995-12-12 | Sclavo Spa | PROCESS FOR THE EXTRACTION OF THE FACTOR VIII-FACTOR VON WILLEBRAND (FVIII: C-FVW) FROM TOTAL HUMAN PLASMA. |
DE4242863A1 (en) * | 1992-12-18 | 1994-06-23 | Boehringer Mannheim Gmbh | Stable lyophilized pharmaceutical preparations of G-CSF |
WO1994017834A1 (en) | 1993-02-09 | 1994-08-18 | Octapharma Ag | Method for inactivating viruses devoid of lipid envelopes |
SE9301581D0 (en) * | 1993-05-07 | 1993-05-07 | Kabi Pharmacia Ab | PROTEIN FORMULATION |
SE504074C2 (en) | 1993-07-05 | 1996-11-04 | Pharmacia Ab | Protein preparation for subcutaneous, intramuscular or intradermal administration |
US5576291A (en) | 1993-09-13 | 1996-11-19 | Baxter International Inc. | Activated factor VIII as a therapeutic agent and method of treating factor VIII deficiency |
US5353835A (en) | 1993-09-23 | 1994-10-11 | Ingersoll-Rand Company | Air tank drain |
IT1268920B1 (en) | 1994-03-29 | 1997-03-13 | Syfal Srl | ROTARY MACHINE FOR DECORATION-GLAZING IN PARTICULAR CERAMIC TILES. |
IL113010A0 (en) | 1994-03-31 | 1995-10-31 | Pharmacia Ab | Pharmaceutical formulation comprising factor VIII or factor ix with an activity of at least 200 IU/ml and an enhancer for improved subcutaneous intramuscular or intradermal administration |
US5514781A (en) | 1994-04-11 | 1996-05-07 | Bayer Corporation | Use of azoles as virucidal agents in solutions of biologically active proteins |
US5955448A (en) | 1994-08-19 | 1999-09-21 | Quadrant Holdings Cambridge Limited | Method for stabilization of biological substances during drying and subsequent storage and compositions thereof |
FR2719479B1 (en) * | 1994-05-04 | 1996-07-26 | Sanofi Elf | Stable lyophilized formulation comprising a protein: assay kit. |
DE69530196T2 (en) | 1994-06-02 | 2003-11-20 | Elan Drug Delivery Ltd | METHOD FOR PREVENTING THE AGGREGATION OF PROTEINS / PEPTIDES IN REHYDRATION OR THAWING |
US5580856A (en) | 1994-07-15 | 1996-12-03 | Prestrelski; Steven J. | Formulation of a reconstituted protein, and method and kit for the production thereof |
DE4431833C1 (en) | 1994-09-07 | 1995-05-18 | Blutspendedienst Der Drk Lande | Prepn. of an anti-haemophilic factor from a cryo-precipitate |
GB2293100A (en) | 1994-09-15 | 1996-03-20 | Medeva Europ Ltd | Pharmaceutical compositions with deuterium oxide |
SE9403915D0 (en) | 1994-11-14 | 1994-11-14 | Annelie Almstedt | Process A |
SE503424C2 (en) | 1994-11-14 | 1996-06-10 | Pharmacia Ab | Process for purification of recombinant coagulation factor VIII |
AU4526996A (en) * | 1994-12-28 | 1996-07-19 | Biotime, Inc. | Plasma expanders and blood substitutes |
GB9501040D0 (en) | 1995-01-19 | 1995-03-08 | Quadrant Holdings Cambridge | Dried composition |
SE9501189D0 (en) * | 1995-03-31 | 1995-03-31 | Pharmacia Ab | Protein formulation |
US5679549A (en) | 1995-05-04 | 1997-10-21 | Bayer Corporation | Production of recombinant factor VIII in the presence of liposome-like substances of mixed composition |
JP3927248B2 (en) * | 1995-07-11 | 2007-06-06 | 第一製薬株式会社 | HGF lyophilized formulation |
KR0167677B1 (en) * | 1995-08-31 | 1999-02-01 | 김광호 | Memory test system with pattern generator for multi-bit test |
SE9503380D0 (en) | 1995-09-29 | 1995-09-29 | Pharmacia Ab | Protein derivatives |
ES2099678B1 (en) | 1995-11-03 | 1998-02-16 | Grifols Grupo Sa | PROCEDURE FOR THE INACTIVATION OF VIRUSES IN PROTEINS. |
ATE235814T1 (en) | 1995-11-06 | 2003-04-15 | New York Blood Ct Inc | TREATMENT OF RED BLOOD CELLS TO INACTIVATE VIRIA BY USING PATHALOCYANINS AND RED LIGHT |
US5659017A (en) | 1995-11-07 | 1997-08-19 | Alpha Therapeutic Corporation | Anion exchange process for the purification of Factor VIII |
US5925738A (en) | 1995-12-01 | 1999-07-20 | The American National Red Cross | Methods of production and use of liquid formulations of plasma proteins |
US5851800A (en) | 1996-05-14 | 1998-12-22 | Pharmacia & Upjohn Ab | Process for producing a protein |
US6632648B1 (en) | 1996-05-14 | 2003-10-14 | Elan Drug Delivery Limited | Methods of terminal sterilization of fibrinogen |
US5763401A (en) * | 1996-07-12 | 1998-06-09 | Bayer Corporation | Stabilized albumin-free recombinant factor VIII preparation having a low sugar content |
PT2193809E (en) * | 1999-02-22 | 2015-08-24 | Baxter Int | Albumin-free factor viii formulations |
JP4674021B2 (en) | 1999-07-13 | 2011-04-20 | ビオヴィトルム・アクチボラゲット(プブリクト) | Stable factor VIII composition |
US6440414B1 (en) * | 1999-10-01 | 2002-08-27 | Amgen Inc. | Pharmaceutical compositions of fibrinolytic agent |
MXPA06006886A (en) | 2003-12-19 | 2006-09-04 | Novo Nordisk Healthcare Ag | Stabilised compositions of factor vii polypeptides. |
EP1977763A4 (en) | 2005-12-28 | 2010-06-02 | Chugai Pharmaceutical Co Ltd | Antibody-containing stabilizing preparation |
JO3324B1 (en) | 2006-04-21 | 2019-03-13 | Amgen Inc | Lyophilized Therapeutic Peptibody Formulations |
EP2337580B1 (en) | 2008-09-03 | 2012-03-28 | Octapharma AG | Stabilized compositions for recombinantly produced factor viii |
-
2000
- 2000-02-22 PT PT100750199T patent/PT2193809E/en unknown
- 2000-02-22 EP EP06077229.0A patent/EP1820516B1/en not_active Expired - Lifetime
- 2000-02-22 CZ CZ2008-827A patent/CZ307322B6/en not_active IP Right Cessation
- 2000-02-22 EP EP10075019.9A patent/EP2193809B1/en not_active Expired - Lifetime
- 2000-02-22 CN CN200910171121A patent/CN101810854A/en active Pending
- 2000-02-22 DK DK00907322T patent/DK1154796T3/en active
- 2000-02-22 US US09/507,011 patent/US6586573B1/en not_active Expired - Lifetime
- 2000-02-22 RU RU2001125929/15A patent/RU2244556C2/en active
- 2000-02-22 PT PT09075396T patent/PT2130554E/en unknown
- 2000-02-22 PL PL382288A patent/PL203177B1/en unknown
- 2000-02-22 EP EP15154020.0A patent/EP2921180B1/en not_active Expired - Lifetime
- 2000-02-22 DK DK06077229.0T patent/DK1820516T3/en active
- 2000-02-22 ES ES06077229T patent/ES2435141T3/en not_active Expired - Lifetime
- 2000-02-22 ES ES10075019.9T patent/ES2541470T3/en not_active Expired - Lifetime
- 2000-02-22 WO PCT/US2000/040068 patent/WO2000048635A1/en active Application Filing
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2003
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2006
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2007
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2008
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2011
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2012
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2013
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2014
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2015
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2016
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2017
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