WFH Bangkok 2004 Nanotiv – Factor IX Concentrate Plasma-derived, Very High Purity Factor IX Concentrate

WFH Bangkok 2004 Nanotiv – Factor IX Concentrate Nanotiv is a highly pure factor IX concentrate derived from carefully selected donors and extensively tested plasma The Nanotiv manufacturing process is designed to fulfil the highest modern requirements for virus safety by combining two reliable and validated methods of virus elimination The Nanotiv manufacturing process is gentle, in order to preserve biological function of the factor IX, reduce the risk of thrombogenicity and lead to good clinical tolerance

WFH Bangkok 2004 Nanotiv – Rigorously Controlled Plasma Carefully selected donors Controlled donation centres Extensive donation testing –HBsAg –Anti-HCV –Anti-HIV –ALT –Syphilis PCR tests on –Mini pools and –Production pools Inventory holding

WFH Bangkok 2004 Nanotiv Manufacturing Process Cryoprecipitate supernatant plasma Anion exchange chromatography S/D Virus inactivation Affinity chromatography Cation exchange chromatography Virus removal: Nanofiltration Diafiltration Ultrafiltration Sterile filtration Lyophilisation NANOTIV

WFH Bangkok 2004 Nanotiv is purified from cryoprecipitate supernatant plasma by a train of chromatography steps which Concentrate the prothrombin complex factors then Selectively enrich factor IX The first virus inactivation step is solvent/detergent (S/D) The second step of virus removal is nanofiltration Albumin is not added to Nanotiv as a stabiliser Nanotiv is highly pure and free of other proteins and activated factors so that heparin does not need to be added Nanotiv Manufacturing Process

WFH Bangkok 2004 Nanotiv – Nanofiltration

WFH Bangkok 2004 Nanotiv – Viral Safety: Conclusion Nanotiv factor IX fulfils all current requirements for virus safety set out by regulatory bodies such as the Committee for Proprietary Medicinal Products* Two effective steps against lipid enveloped viruses One effective step against non enveloped viruses A combination of methods based on different principles of action Inactivation/removal with a high safety margin Rapid virus inactivation Robustness in the event of process variations Validation of each step with a wide variety of viruses An individual step efficacy equivalent to 4 log 10 *CPMP/BWP/268/95, CPMP/BWP/269/95 rev 3., 2001

WFH Bangkok 2004 Nanotiv Virus Validation Studies Enveloped VirusesNon Enveloped Viruses Virus Genome HIV-1 Human Immuno- deficiency Virus RNA BVDV Bovine Viral Diarrhoe Virus RNA PRV Pseudo- rabies Virus DNA HAV Hepatitis A Virus RNA PPV Porcine Parvovirus DNA StepVirus Reduction (log 10 ) S/D Treatment> 4.9 ± 0.4> 5.8 ± 0.3> 5.4 ± 0.4 Nanofiltration> 4.6 ± 0.2> 6.2> – 4.2 Total Reduction > 9.5> 12.0> – 4.2

WFH Bangkok 2004 Nanotiv Purity Nanotiv is among the purest factor IX preparations available Prothrombin complex factors are removed to trace amounts The high purity of Nanotiv is the prerequisite for its excellent clinical tolerability and efficacy

WFH Bangkok 2004 Alb: Albumin A: Nanotiv B-E: Other commercial factor IX preparations Mr: Molecular weight markers : Factor IX monomer Nanotiv Purity – SDS-PAGE Non reduced Reduced Non reduced Silver-stained Silver-stained Western Blot anti-F IX Abs After: Moberg U, et al. XXII Int. Congr. WFH. Dublin, June 23-28, 1996

WFH Bangkok 2004 Nanotiv Purity – Trace Proteins FactorIU/ml Proportion of Factor IX Factor IX:C1001/1 Factor II< 0.02< 1/5,000 Factor VII< 0.005< 1/20,000 Factor X< 0.2< 1/500 Minimised Prothrombin Complex Factors

WFH Bangkok 2004 Nanotiv Purity – Gel Filtration After: Moberg U, et al. XXII Int. Congr. WFH. Dublin, June 23-28, 1996

WFH Bangkok 2004 High purity factor IX preparations, like Nanotiv are almost devoid of other vitamin K-dependent factors, as well as phospholipids They are less likely to be thrombogenic, compared with products of low purity Even at very high dose (200 IU/kg BW), Nanotiv still does not have an elevated thrombogenic effect Nanotiv – In Vivo Thrombogenicity

WFH Bangkok 2004 Nanotiv 200 IU/kg Control 1 ml saline/kg Highly thrombogenic area PCC 25 IU/kg Nanotiv – In Vivo Thrombogenicity Wessler Rabbit Stasis Model Thrombogenic index (TI)

WFH Bangkok 2004 Nanotiv – Clinical Efficacy, Pharmacokinetics Mean plasma concentration (IU/ml) vs. time (h), n = 12

WFH Bangkok 2004 Nanotiv – Clinical Efficacy Half-life (t½, hours)22.6 In vivo recovery (%)58.3 Incremental recovery (IU/dl per IU/kg)1.2 Clearance (ml x kg x h -1 )4.84 Pharmacokinetic Parameters

WFH Bangkok 2004 Nanotiv – 6 Months Follow Up Study Most bleeding episodes were managed using a single dose of (mean) 29 IU/kg BW Number of injections per bleed vs. Cumulative % of bleeds managed % of bleeds

WFH Bangkok 2004 Nanotiv – Clinical Efficacy Summary In comparative pharmacokinetic studies, Nanotiv half-life and recovery values were within the normal range After 6 months, the same cohort of patients showed similar values for half-life and recovery as at the beginning Nanotiv gave no inhibitors Most bleeding episodes were managed using a single dose of Nanotiv with a mean of 29 IU/kg BW Nanotiv was well-tolerated

WFH Bangkok 2004 Nanotiv – Clinical Efficacy in Surgery Nanotiv was well-tolerated in surgery At high bolus doses and in continuous infusion The haemostatic effect always rated as good or excellent No signs of thrombosis or pulmonary embolism observed No adverse events observed No inhibitors detected 14 days after conclusion of continuous infusion

WFH Bangkok 2004 Nanotiv – Clinical Efficacy in Surgery CaseProcedure Days CI Total Dose (IU) Blood Loss Trans- fusion Haemostasis 1Total hip replacement NormalNoExcellent 2Disc removal L5-S1 left NormalNoExcellent 3Ankle arthrodesis NormalNoExcellent 4Pseudoarthrosis resection NormalNoExcellent 5Total knee replacement NormalNoExcellent 6Radial head resection and arthrolysis NormalNoExcellent 7Total knee replacement NormalNoExcellent 8Appendectomy NormalNoExcellent 9Port catheter insertion NormalNoExcellent 10Circumcision NormalNoExcellent 11Liver biopsy NormalNoExcellent 12Total knee replacement NormalNoExcellent 13Liver biopsy NormalNoExcellent Schulman S, et al. Haemophilia 1999; 5:

WFH Bangkok 2004 Nanotiv – Convenience: All You Need Water for Injections Alcohol swab x 3 Transfer needle Filter needle Syringe Infusion line (Butterfly) Gauze Plaster

WFH Bangkok 2004 Nanotiv at a Glance – Key Features Highly purified factor IX Triple chromatography process Specific activity ~ 200 IU/mg Gentle purification Non denatured Heparin-free formulation Reduced risk of thrombocytopenia Minimal interference with diagnostic tests Complementary virus safety concept High quality starting plasma Double virus elimination procedures Solvent / Detergent (S/D) treatment Nanofiltration Chromatographic steps provide additional safety

WFH Bangkok 2004 Nanotiv at a Glance – In Daily Clinical Use Extensive clinical experience Excellent general safety proven over more than 10 y Effective Well-tolerated Convenient handling Small injection volume Easy documentation Pull-off labels Dissolves rapidly Presentation Storage and shelf-life 36 months shelf-life at + 4 °C to + 8 °C 1 month storage at elevated temperature up to 25 °C Package Size (IU)Injection Volume Nanotiv 5005 ml Nanotiv ml