Classifications

• • 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
  • A61K38/43—Enzymes; Proenzymes; Derivatives thereof
  • A61K38/46—Hydrolases (3)
  • A61K38/48—Hydrolases (3) acting on peptide bonds (3.4)
  • A61K38/482—Serine endopeptidases (3.4.21)
  • A61K38/4846—Factor VII (3.4.21.21); Factor IX (3.4.21.22); Factor Xa (3.4.21.6); Factor XI (3.4.21.27); Factor XII (3.4.21.38)
• • 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

Definitions

• This invention relates to the treatment of haemophilia and is particularly concerned with the administration of factor IX.
• Factor IX is a component of the blood-clotting mechanism of warm-blooded animals : it is Involved together with many other blood clotting factors in the blood coagulation cascade as reviewed by Furie B. and Furle B. C, Cell, 53, 1988, pages 505-518.
• Haemophilia B, or Christmas disease is a serious inherited bleeding disorder affecting males and caused by a defect in clotting factor IX. Control of the disease is effected by the need for repeated injections of factor IX concentrate, conventionally obtained from blood donors, although more recent technology (as exemplified by EP-A-195592) allows its preparation by recomblnant DNA technology.
• Factor IX is known to be a single-chain glycoprotein with a molecular weight of approximately 60,000. It is therefore a sufficiently large molecule to point to the need for direct introduction into the bloodstream. Furthermore quite large doses of factor IX (e.g. of the order of 5,000 international, units) may be required for an adult, for example during surgery. Consequently, factor IX preparations have conventionally been delivered intravenously to patients, either prophylactlcally or in response to bleeding episodes in order to control haemophilia B.
• factor IX can be delivered by subcutaneous injection with sufficiently rapid transport into the bloodstream in biologically active form and in adequate concentrations for control of haemophilia B.
• the present invention provides a kit for use in subcutaneous injection comprising factor IX and a pharmaceutically acceptable carrier adapted for delivery of an effective dose of factor IX to a haemophilia B patient by such subcutaneous injection.
• the invention also provides factor IX for use in the preparation of a composition for subcutaneous injection of factor IX to a haemophilia B patient.
• the invention further provides a method of treatment of a haemophilia B patient comprising subcutaneous injection of the patient with a composition comprising factor IX.
• the factor IX employed in the invention may be in the form of a concentrate obtained from blood donors or may have been obtained by recombinant DNA technology.
• the factor IX is preferably human factor IX for the treatment of human haemophilia B patients, although in principle the invention can be applied to the introduction of human or animal factor IX to other warm-blooded animals with an analogous blood clotting mechanism.
• the factor IX may be initially supplied as, or made up as, a composition in a pharmaceutically acceptable carrier such as deionised water or physiological saline.
• solid factor IX in suspension may be employed for administration subcutaneously, allowing slow release into the bloodstream from the site of injection.
• Other comparatively low molecular weight additives and excipients may be present.
• the factor IX is preferably substantially free of other blood clotting factors such as flbrinogen, prothrombin, factors VII, VIII, X and XI which may be present in some commercially available sources of factor IX.
• other blood clotting factors may be removed by standard purification procedures such as passing over an immuno-afflnity column loaded with a monoclonal antibody specific for factor IX as described by Rees et al., EMBO J., 7, 1988, pages 2053-2061.
• the factor IX conveniently in lyophilised form, and any carrier may be provided separately in a kit with the intention of combining the factor IX and carrier Immediately prior to use. Instructions for calculating the dosage to be delivered dependent on the patient's body weight and the circumstances of injection and appropriate for subcutaneous injection will conventionally be included in such a kit.
• the factor IX can be delivered subcutaneously either prophylactically or in response to bleeding episodes. It will be apparent that the use of subcutaneous delivery is particularly valuable for prophylaxis where there is a lower dose requirement.
• Lyophilised human factor IX (available as "Mononine” from Armour Pharmaceutical Company, Kankakee, II., USA) was dissolved in deionised water at concentrations ranging from 10 to 500 international units (i.u.) per ml.
• Fig. 1 shows the results and indicates that factor IX is transported into the bloodstream via subcutaneous injection and reaches a concentration which is approximately 40% of the concentration reached in plasma after intravenous injection of an equivalent amount of factor IX.
• Factor IX in the purified samples was quantified by ELISA for antigen essentially as described by Anson et al., Nature, 315, 1985, pages 683-685.
• Factor IX clotting activity was quantified by the one-step clotting assay of Austen and Rhymes, A Laboratory Manual of Blood Coagulation, 1975, Blackwell Scientific Oxford, page 59. In both cases, pooled normal human plasma was used as the standard.
• Lyophilised human factor IX as used in Examples 1 and 2 was treated to remove any low molecular weight additives as follows:
• Step 1 20 i.u. factor IX were dissolved in 2ml delonised water.
• Step 2 The solution was mlcroconcentrated by centrlfugation for 1 hour and the resulting solution, after an estimated 37% loss (estimated by ELISA), contained 12.6 i.u. in a volume of 85 ⁇ l. Although the concentration of additives in this factor IX solution remains unchanged from that in the initial solution, they were reduced in amounts by a calculated 23.5 fold (i.e. 2000 ⁇ l ⁇ 85 ⁇ l).
• Step 3 The factor IX solution (12.6 i.u. in 85 ⁇ l) was then diluted to 1ml with physiological saline, to give a solution containing 12.6 i.u. factor IX/ml.
• mice Two mice were injected intravenously into the tail vein and two mice were injected subcutaneously into the back. In each case the total volume of 100 ⁇ l was injected into a single site.
• 0.1-0.2 ml blood samples were taken from the tail vein at intervals over two days and the human factor IX in the plasma quantified by ELISA as described above. The results are shown in Fig. 3 and are comparable with those obtained in Figs. 1 and 2 showing that the presence of low molecular weight additives is not a significant contributory factor to the ability of the factor IX to reach the bloodstream.
• factor IX Factor IX
• IXMC available from Bio Products Laboratory, Elstree, Herts, UK
• concentration of 81 international units/ml as determined by ELISA
• the preparation was believed to further contain low molecular weight additives.
• 8.1 international units factor IX in 100 ⁇ l was injected into each of four mice. Two mice were injected intravenously into the tail vein and two mice, were injected subcutaneously into the back. In each case the total volume was injected into a single site. 0.1-0.2 ml blood samples were taken from the tail vein at intervals over two days and the human factor IX in the plasma was quantified by ELISA as described above. The results are shown in Fig. 4 and show that factor IX is transported to the bloodstream from a subcutaneous injection site with an efficiency of transport of 25% and 43% depending on the mouse (comparing the results of the subcutaneously injected mice with the average of the two intravenously injected mice).
• Factor IXA Factor IXA (BPL) 585 i.u./bottle.
• Reconstituted solution when dissolved in 20ml H 2 O contains per litre not more than: 20g protein, 300 mMoles sodium, 200 mMoles chloride, 50 mMoles phosphate, 60 mMoles citrate, 500 i.u. antithrombin III, 5000 u. heparin; nominal content of factor II 800 u.; factor X 400 u.; factor VII negligible for therapeutic purposes.
• the recommended concentration for human use (i.v.) was 585 i.u. in 20ml.
• Factor IX was dissolved to a concentration of 585 i.u. in 10ml of deionised water. An ELISA assay showed that this product in fact contained 89.4 international units/ml (447 ⁇ g/ml).

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• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Diabetes (AREA)
• General Chemical & Material Sciences (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Hematology (AREA)
• Gastroenterology & Hepatology (AREA)
• Immunology (AREA)
• Proteomics, Peptides & Aminoacids (AREA)
• Epidemiology (AREA)
• Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
• Treatments Of Macromolecular Shaped Articles (AREA)

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• 1991
  • 1991-10-24 GB GB919122609A patent/GB9122609D0/en active Pending
• 1992
  • 1992-10-20 GB GB9222012A patent/GB2260702B/en not_active Expired - Fee Related
  • 1992-10-20 JP JP5507561A patent/JPH07502989A/ja active Pending
  • 1992-10-20 EP EP92921475A patent/EP0609293A1/en not_active Withdrawn
  • 1992-10-20 WO PCT/GB1992/001926 patent/WO1993007890A1/en not_active Application Discontinuation
  • 1992-10-20 AU AU27720/92A patent/AU2772092A/en not_active Abandoned

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