Eribis 2011

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  • Bullets Founded 2006 Cardiovascular disease Ventuer cap
  • Ref: veronique L Roger, 2007
  • PCI slide shows stewnt thrombosis Percutaneous Coronary Intervention ( PCI ) ACC/AHA/SCAI Guidelines A joint 2005 American College of Cardiology/ American Heart Association/ Society for Cardiovascular Angiography and Interventions report gives guidelines for the management of patients undergoing PCI.  The report strongly recommends that PCI should be performed in facilities that have an experienced cardiovascular surgical team available as emergency backup for all procedures. 1. Peters RJG, et al. BMJ . 2007;334:1265-1269. 2. Gibler WB, et al. Circulation . 2005;111:2699-2710. 3. Medi C, et al. Med J Aust . 2007;186:197-202.
  • Small IPost non-pharmacologic clinical studies: Staat P, Circulation 2005 STEMI, no signs of spontaneous reperfusion, reperfusion <6 h after onset chest pain IPost protocol consisted of 4 cycles of 1-min inflation of angioplasty baloon, applied within 1 min of reopening of the vessel a 36% reduction of AUC for serum CK release was reported Provided proof of concept Thibault H, Circulation, 2008 Confirmed long-term benefit of IPost in a similar design as the Staat study 2005, except that, SPECT and ECHO were performed 6 and 12 months after PCI Found a 40% reduction in infarct size (enzymes and SPECT) and improved LVEF and wall motion score index (WMSI) Studies by Laskey WK (2005, 2008), Ma X, 2006 confirmed the above studies
  • ADME is an acronym in pharmacokinetics and pharmacology for a bsorption, d istribution, m etabolism, and e xcretion, and describes the disposition of a pharmaceutical compound within an organism. The four criteria all influence the drug levels and kinetics of drug exposure to the tissues and hence influence the performance and pharmacological activity of the compound as a drug
  • Safety Treatment-emergent events Leading to premature discontinuation SAEs
  • Safety Treatment-emergent events Leading to premature discontinuation SAEs
  • Eribis 2011

    1. 1. Eribis Pharmaceuticals AB Novel Treatment of Acute Myocardial Infarction (AMI)CONFIDENTIAL 1
    2. 2. Eribis presentation  The Company  The indication – background & market  EP94  Pre-clinical Development  General Properties  Clinical Development  Intellectual Property  Q & A sessionCONFIDENTIAL 2
    3. 3. The CompanyCONFIDENTIAL 3
    4. 4. The Company Eribis Pharmaceuticals AB is a Uppsala-based biotech company founded in 2006, with the aim to develop new therapies for cardiovascular disease.CONFIDENTIAL 4
    5. 5. Management  Herman Krapf, Chief Executive Officer  Erik Bissessar, Chief Business Officer and Medical Scientific Liason  Peter Båvenholm, MD, Ph.D., Ass. Prof. Internal Medicine, Chief Medical Officer  Fredrik Röök, Business Development  Stefan Persson, Ph.D., Pharmacology and Toxicology  Claes Lundberg, Ph.D., Project ManagementCONFIDENTIAL 5
    6. 6. Scientific Advisory Board  Gerhard Wikström, MD, Ph.D., Ass Prof Cardiology, Uppsala University Hospital  Lars Grip, MD, Ph.D., Professor in Cardiology, Sahlgrenska University Hospital  John Pernow, MD, Ph.D., Prof in Cardiology, Karolinska University Hospital  Garrett Gross, Ph.D.,Professor of Pharmacology, Medical College of Wisconsin  Dan Atar, MD, Ph.D., Professor in Cardiology, Division of Cardiology, Oslo University Hospital AkerCONFIDENTIAL 6
    7. 7. Acute Myocardial Infarction (AMI) background and marketCONFIDENTIAL 7
    8. 8. Cardiovascular disease (CVD) the major health problem  CVD include;  acute coronary syndromes (ACS)  stroke  periphereal arterial disease (PAD)  Burden of CVD is increasing in parallel with increase in life expectancy  Acute Myocardial Infarction (AMI) is the number one cause of death in the Western world  mortality rates near 10%  first cause of chronic heart failureCONFIDENTIAL 8
    9. 9. Acute coronary syndrome (ACS) Acute coronary syndrome (ACS) is comprises three diseases involving the coronary arteries: ST elevation myocardial infarction (STEMI), non ST elevation myocardial infarction (non-STEMI) and unstable angina. While ACS mortality declined in the last four decades in the USA as life expectancy increased, the decline largely represents the postponement of ACS deaths until older age. Thus, the burden of ACS is increasing in parallel with the increase in life expectancy.CONFIDENTIAL 9
    10. 10. Reperfusion therapy Primary Treatment O ptions for AMI Coronary Artery By-pass Graft (CABG) Percutaneous Coronary Intervention (PCI) Across the seven major markets in 2006 the number of cardiovascular procedures, including CABG and PCI were estimated at 7.0m Incidence rates for PCI amounts to 3.7m (nonSTEMI + STEMI) 10
    11. 11. Large unmet need Acute Coronary Syndrome - the major markets The American Heart Association (AHA) estimates that there were 1.2 million cases of ACS in 2007 in the US. This number includes both patients experiencing their first event as well as patients with relapses. Patients’ experiencing their first event is around 700,000; STEMI and NSTEMI accounting for 85% of these events. This incidence is similar to that in the five major markets in Europe. The total number of ACS cases in the US and the five major EU markets is therefore estimated at around 2.5 – 3.0 million annually.CONFIDENTIAL 11
    12. 12. Metabolic and biochemic changes caused by reperfusion Reperfusion injury (RI) limits the beneficial effects of revascularisation Studies in different species suggest that 20-70% of myocardial damage may come from RI (Gomes L 2007, Penna C 2008, Zhao ZQ, 2006) RI leads to functional and structural myocardial damage –generation of ROS –a fast restoration of pH –intracellular calcium overload –mitochondrial dysfunction –Apoptosis (Yellon DM, 2007) Cardioprotective strategies to minimize RI represents an unmet medical needCONFIDENTIAL 12
    13. 13. Principles of ischemic pre- and postconditioning Brief episodes of coronary artery occlusion are applied either before (preconditioning) or immediately after (postconditioning) the prolonged ischemic insult CONFIDENTIALRef; Zhao ZQ et al., Am J Physiol Heart Circ Physiol, 2003;285:579-88 13
    14. 14. EP94 Cardioprotection in acute myocardial infarction (AMI)CONFIDENTIAL 14
    15. 15. EP94 – First in class drug  Novel pharmacological cardioprotective intervention  Tetrapeptide (stabilized)  Injectable, small volume  Microgram dosage  Cardioprotective effects in rodent and non-rodent animal models  Low COGS (solid phase peptide synthesis)  Patent protectedCONFIDENTIAL 15
    16. 16. Tentative molecular signaling pathways involved in cardioprotectionCONFIDENTIAL 16
    17. 17. Mode of Action  Acute Myocardial Infarction induces an ischemia/reperfusion-induced tissue damage  It is anticipated that EP94’s mode of action reduces the ischemia/reperfusion damage through  opoid receptors  KATP channels  iNOSCONFIDENTIAL 17
    18. 18. Preclinical resultsCONFIDENTIAL 18
    19. 19. Key studies presented: • Rodent dose finding study • Pig study, closed chest, iv. • Pig study, open chest, infusion • Pig study, closed chest, dose finding, iv. • Pig study, closed chest, high-dose range finding, iv. • Rodent study, dose finding, m.o.a. studyCONFIDENTIAL 19
    20. 20. EP94 shows a dose dependent reduction of myocardial infarction size in rats 25 min occlusion = Vehicle administration -5 0 25 = EP94 administration Vehicle EP94 (0.01 µg/kg x 1) EP94 (0.1 µg/kg x 1) Post-surgery 2 hrs reperfusion stabilization EP94 (1 µg/kg x 1) phase EP94 (5 µg/kg x 1) EP94 (10 µg/kg x 1) 60 50 40 * * * IS/AAR (%) 30 *Pre-conditioning 20 10 N=12 N=6 N=12 N=12 N=12 N=9 0 le k g) /kg) / k g) /kg) /kg ) V ehic (0.01 ug/ .1 ug P94 (1 ug P94 (5 ug 94 (10 ug EP9 4 EP 94 (0 E E EP CONFIDENTIAL Study number 64-06 20
    21. 21. EP94 i.v. administered early or late during ischaemia in a closed chest MI pig model reduces myocardial infarction size significantly 40 min occlusion = Vehicle administration 60 min 5 12 19 26 33 40 = EP94 administration Vehicle Post-surgery EP94 (1 µg/kg x 4) – Early intervention stabilization 4 hrs reperfusion phase EP94 (1 µg/kg x 3) – Late intervention EP94 (0.2 µg/kg x 3) – Late intervention Closed chest pig MI/reperfusion model 80 70 ** 60 * IS/AAR (%) 50 40 30 Early Late Late 20 10 N =7 N =4 N =6 N =6 0 hic le x 4) x 3) x 3) Ve / kg /kg / kg ug ug ug (1 (1 (0. 2 94 94 94 EP EP EP Mean ± SEMCONFIDENTIAL Study Report 09-01, Karlsson et al., Eur J Pharmacol 2011:651 pp146-151 21
    22. 22. Intracoronary infusion of EP94 in pigs reduces myocardial infarction size significantly 30’ 5’ 60 min 40 min occlusion Infusion Post-surgery Vehicle stabilization 4 hrs reperfusion phase EP94 (0.2 µg/min, 15 min) 100 Open chest pig MI/reperfusion model 90 80 ** % of area at risk 70 60 50 40 30 20 10 N=5 N=5 0 Mean ± SEM Vehicle EP94CONFIDENTIAL 22 Study Report 08-02, Karlsson et al., Eur J Pharmacol 2011:651 pp146-151
    23. 23. EP94 appears to reduce myocardial infarction size dose-dependently 40 min occlusion = Vehicle administration 60 min 26 33 40 = EP94 administration Vehicle Post-surgery EP94 (1 µg/kg x 3) stabilization 4 hrs reperfusion phase EP94 (5 µg/kg x 3) EP94 (25 µg/kg x 3) Closed chest pig MI/reperfusion model 70 60 50 IS/AAR (%) 40 30 20 10 N=6 N=6 N=6 N=6 0 Vehicle 1 ug/kg 5 ug/kg 25 ug/kg Mean ± SEMCONFIDENTIAL Study Report 11-02 23
    24. 24. Ongoing studies - preliminary resultsCONFIDENTIAL 24
    25. 25. Dose range finding study in a closed chest pig model 40 min occlusion = Vehicle administration = EP94 administration 60 min 0 26 33 40 Vehicle EP94 (1 µg/kg x 3) Post-surgery EP94 (25 µg/kg x 3) stabilization 4 hrs reperfusion phase EP94 (125 µg/kg x 3) OPTIONAL: EP94 (625 µg/kg x 3) N = 12/groupCONFIDENTIAL Study protocol 2011-01 25
    26. 26. Results of the dose finding study is a…CONFIDENTIAL 26
    27. 27. Rat acute myocardial infarction model – a combined dose response and mode of action study 25 min occlusion = Vehicle administration -10 0 5 10 = EP94 administration Vehicle EP94 (0.5 µg/kg x 2) Post-surgery EP94 (1.0 µg/kg x 2) stabilization 2 hrs reperfusion phase EP94 (2.5 µg/kg x 2) EP94 (5.0 µg/kg x 2) EP94 (10 µg/kg x 2)1) Opioid antagonists i.v. 2) KATP channel antagonists i.v. 3) iNOS inhibitor i.v. a) Naltrindole (delta) – 5 mg/kg a) HMR1098 (sarc KATP blocker) – 6 mg/kg a) 1400W b) Nor-BNI (kappa) – 0.3 mg/kg b) 5-HD (mito KATP blocker) – 10 mg/kg c) CTOP (mu) – 0.1 mg/kg b) 1400W + EP94 d) HMR1098 + EP94 d) Naltrindole + EP94 e) 5-HD + EP94 e) Nor-BNI + EP94 f) CTOP + EP94CONFIDENTIAL Study protocol 2010-01 27
    28. 28. Rat acute myocardial infarction model – a combined dose response and mode of action study - cont´ d Bell-shaped dose-response relationship in rat confirms previous results in rats 70,00 60,00 ** 50,00 *** *** IS/AAR (%) 40,00 30,00 20,00 10,00 N=9 N=8 N = 10 N = 10 N=8 N=9 N=8 0,00 Vehicle 0.1 0.5 1.0 2.5 5.0 10.0 EP94 (ug/kg, i.v.) Mean ± SEM *** p < 0.001 vs vehicle ** p < 0.01 vs vehicleCONFIDENTIAL 28
    29. 29. Rat acute myocardial infarction model – a combined dose response and mode of action study - cont´ d Blocking of mu-opioid receptors abolish the cardioprotective effects of EP94 70,00 60,00 ** ** 50,00 *** IS/AAR (%) 40,00 30,00 20,00 10,00 0,00 N=9 N=9 N=9 N=9 N = 10 N=9 N=9 N=9 ) ) ) ) 4 4 c le kg kg kg kg P9 94 P9 hi g/ g/ g/ g/ E EP E Ve m m m u + + I+ (5 (0 .1 0. 3 (1 NT O P BN NT I( 94 CT r- O P N EP no CT r-B No NT = naltrindole = selective delta-antagonist Mean ± SEM CTOP = selective mu-antagonist *** p < 0.001 vs vehicle Nor-BNI = selective kappa-antagonist ** p < 0.01 vs vehicleCONFIDENTIAL 29
    30. 30. Rat acute myocardial infarction model – a combined dose response and mode of action study - cont´ d Effects of selective KATP ion channel antagonists 70,00 60,00 50,00 *** IS/AAR (%) 40,00 30,00 20,00 10,00 N=9 N = 10 N=9 N=9 N=9 N=9 0,00 le kg ) g) 4 g) 94 hi c g/ g/ k P9 g/ k E EP Ve (1 u m + 0 m + (6 98 94 98 10 (1 HD HD 5- EP 10 R 5- R HM HM Mean ± SEM HMR 1098 = sarcolemmal KATP channel antagonist *** p < 0.001 vs vehicle 5-HD = mitochondrial KATP channel antagonistCONFIDENTIAL 30
    31. 31. Rat acute myocardial infarction model – a combined dose response and mode of action study - cont´ d Effects of iNOS inhibition on EP94-induced cardioprotection 70,00 60,00 50,00 *** *** IS/AAR (%) 40,00 30,00 20,00 10,00 0,00 c le 24- h u te e) e) Vehi - ) - Ac (ac ut (ac ut ipx 2) /kg iv EP94 4 00W ug/ kg mg e) + )+1 EP9 4 (1 (0. 1 (ac ut (24- h 14 00W 14 00W E P9 4 Mean ± SEM n = 9/group 1400W = iNOS inhibitor *** p < 0.001 vs vehicleCONFIDENTIAL 31
    32. 32. Conclusions • EP94 is effective in two different pig ischemia/reperfusion models (open vs closed chest) • EP94 produces a significant reduction of infarct size in pigs already at 1 µg/kg • The cardioprotective effect of EP94 seems to be dose- dependent in pigs • EP94 reduces the infarct size dose-dependently during both pre- and post-conditioning in rats • EP94 acts predominantly through mu-receptors in ratsCONFIDENTIAL 32
    33. 33. General properties, Toxicology and Safety PharmacologyCONFIDENTIAL 33
    34. 34. EP 94 – General properties  Tetrapeptide  Manufactured by solid phase peptide synthesis  Lyophilized powder  Solubility: Good solubility in water  Stability of drug substance:  Stable at +4°C for more than 2 years  Stability of drug product:  Stable at -18°C, +4°C and 25°C for more than 6 months in salineCONFIDENTIAL 34
    35. 35. Toxicology & Safety  Single dose tox. study in rats and pigs  Repeat dose tox. study, 14 days, in rats and pigs  Safety Pharmacology  Genotoxicity  Pharmacokinetics and ADMECONFIDENTIAL 35
    36. 36. Clinical DevelopmentCONFIDENTIAL 36
    37. 37. Key factors for success with EP94 in a clinical development programme  Selection of patient population  STEMI patients  An adequate risk stratification algoritm  Selection of primary endpoint  Optimal method used to detect infarct size reductionCONFIDENTIAL 37
    38. 38. Clinical development of EP 94 Phase I study  Study Objective: Dose-escalation study to evaluated the safety, and tolerability in healthy individuals  Design: single-center, double-blind  Estimated number of healthy volunteers: 40  Primary outcome measures: tolerability, safety, PK and hemodynamic parametersCONFIDENTIAL 38
    39. 39. Clinical development of EP 94 Phase II, proof-of-concept study  Study Objective: Dose-escalation study to investigate the cardioprotective effect of ERIBIS Peptide 94 given as an adjunct to percutaneous coronary intervention (PCI) in subjects with an acute STEMI  Design: randomized, placebo-controlled, double-blind and multicenter  Estimated number of patients: 350-400  Primary outcome measure:  To demonstrate a dose-dependent positive trend for myocardial salvage and myocardial salvage index using a modified single contrast enhanced steady-state free precession (SSFP) cine cardiovascular magnetic resonance (CMR) examination performed one week after the acute eventCONFIDENTIAL 39
    40. 40. Clinical development of EP 94 Phase II, proof-of-concept study  Secondary outcome measure:  ST-resolution on 12 lead ECG 90 minutes after PCI  Tropinin, CK-MB 3 to 6 hrs, 6 to 12 hrs, 18 to 24 hrs and 36 hrs after randomization (Peak and AUC)  To determine Left Ventricular Ejection Fraction (LVEF) and Wall Motion Score Index (WMSI) 6 weeks and 6 months after PCI  Myocardial salvage index 6 months after PCI  Incidence of cardiac death and total mortality, stroke, new-onset heart failure, and re-hospitalization for any congestive heart failure 6 month follow-up visitCONFIDENTIAL 40
    41. 41. Intellectual PropertyCONFIDENTIAL 41
    42. 42. Intelectual Property The IPR comprises patent protection for:  Chemical structure of peptides and peptide-based compounds  Clinical Utility  Pharmaceutical Composition  PCT application was filed in August 2007  Regional and National patent applications submitted early 2009 in EU, USA, Canada, Australia, India, China and Japan.  EU patent grant subject for approval, EPO decision to grant obtained Feb 2011CONFIDENTIAL 42
    43. 43. Summary  First in class  Significant cardioprotective effect  Clear unmet medical need & large market size  New therapeutic principle  Significant preclinical results - good clinical prognosisCONFIDENTIAL 43
    44. 44. Q & A answers1. Does the peptide pass the blood brain barrier?2. Does the NO-group give rise to worries in relation totoxicology?3. Does the NO-group give rise to worries in relation toCarcinogenicity?4. Are other receptors hit by this peptide?CONFIDENTIAL 44
    45. 45. NO-study EP94 has been evaluated with and without NO and acylation in response to the EU patent office’s questions regarding Eribis application. To show that the specific groups at position 2, acylation, and pos 4, NO- group, are both necessary to have the cardioprotective effect shown with EP94. 5 different peptides were compared with EP94 in this pre-conditioning study. •without NO-group of the 4th amino acid •without acylation of 2nd amino acid •without both acylation and NO-group •with Asp instead of acylation on 2nd amino acid •with Arg-Asp instead of acylation on 2nd amino acid EP94 does not exert a significant level of cardioprotective activity without these specific groups in the second and fourth position on EP94.CONFIDENTIAL 45
    46. 46. Do you have rodent data? How does EP94 and morphine perform together in the models? Any efficacy measures/differentiation from morphine/other opioids. Answer: Eribis has performed a series of studies on rodents that answer these questions. This data will be available when Zealand performs a DD. There is too much data to discuss during this initial meeting.CONFIDENTIAL 46
    47. 47. Morphine cont’ Morphine is regularly used as pre-medications in the pig studies. EP94 exerts its cardioprotective properties despite approx 13-333-fold higher morphine doses in pigs. However, morphine has not regularly been used as pre-medication in the explorative rat efficacy (cardioprotection) studies conducted in Moscow. MI patients are usually treated with morphine in the emergency room to reduce pain and anxiety. Thus, one exploratory study in rats has been conducted to address the possibility of an interaction between EP94 and morphine. This study did not reveal any synergistic, additive or antagonistic effects of morphine on EP94-induced cardioprotection. Consequently, EP94 has been used in combination with morphine in the pig studies to mimic the clinical situation as closely as possible. The pigs are routinely premedicated with 0.5 mg/kg/hr of morphine.CONFIDENTIAL 47
    48. 48. How is the peptide cleared? Answer: It is anticipated that EP94 is degraded by various endo- and exo-peptidases in the blood stream. The degradation products are believed to be excreted in the urine or partly utilized by the body. This will be verified by dedicated ADME studies in the near future.CONFIDENTIAL 48
    49. 49. Have you tried the peptide, but without the NO-group, in your models? Data from Rodents (cardio protective effect with EP94 (with and without NO) - Y see study 257/10 es How does nitroglycerin perform in your models? - Not evaluated How does nitroglycerin and morphine perform together in the models? (against EP94) - Not evaluated Models where EP94 does not work (MOA)? - EP94 has been shown to be cardioprotective in all models tested so far, i.e. 2 different species and in total 5 different models.CONFIDENTIAL 49
    50. 50. Any cellular assays (all in vitro receptor data (cell lines over- expression the receptors) and work performed on primary cells and specific cell lines that express the receptors). Receptor binding to the human opioid receptors (µ, δ, κ) expressed on CHO and HEK-293 cells. - Study available during the DD process.CONFIDENTIAL 50
    51. 51. Speculations on down stream target? To investigate a possible downstream pathway which may be mediating the beneficial effect of EP94 we administered either the sarcolemmal KATP channel antagonist, HMR 1098, or the mitochondrial KATP channel antagonist, 5-HD, to rats prior to EP94 administration. Both antagonists completely abolished the cardioprotective effect of EP94 which suggests an important role for the KATP channel as a key mediator in the pathway by which EP94 reduces infarct size similar to that seen with a number of other cardioprotective agents as well as ischemic preconditioning and postconditioning.CONFIDENTIAL 51
    52. 52. Are the coronary arteries also affected? Potential effects on coronary arteries have not been investigated.CONFIDENTIAL 52
    53. 53. Speculations on general cardiovascular safety of EP94. • EP94 does not affect blood pressure or heart rate in anaesthesized animals (rats and pigs) at doses levels ranging from 20 – 75 µg/kg (cf. pharmacological efficacy studies) • EP94 does not have significant effects on blood pressure in and only a minor and late effect on heart rate in consious rats at doses of 5-10 mg/kg (ESR08-006). • The very low doses needed for pharmacological efficacy and the short t1/2 (approx 40 min) is anticipated to reduce cardiovascular safety risks to a minimum.CONFIDENTIAL 53
    54. 54. " In-vivo SAR" (opioid component vs. the nitric oxide effects). Unknown Toxicology: Metabolism/metabolites and their distribution. No evaluation of metabolites has been performed. It is anticipated that the bond between amino acid 2 and 3 is susceptible to proteolytic cleavage and that the major metabolites will contain 2 amino acids each. ADME will be performed in the near future.CONFIDENTIAL 54
    55. 55. A list of all published patent applications and patents and their status. PCT application PD53743CA00 - Novel Enkephalin Analogues - Eribis Pharmaceuticals AB EU pat appl. No 07 794 118.5 A “Decision to Grant” will be announced in Q2-2011 Eribis Pharmaceuticals AB owns the IP rightsCONFIDENTIAL 55
    56. 56. FTO analysis of key IP FTO analysis has not been performed, however patentability has been evaluated by third party patent attorneys. - statements are in the handouts.CONFIDENTIAL 56
    57. 57. Thank you for listening! Contact; Herman Krapf, CEO herman.krapf@ eribispharma.se Erik Bissessar Founder & Chief Business Officer erik.bissessar@ eribispharma.seCONFIDENTIAL 57
    58. 58. Back-up bilderCONFIDENTIAL 58
    59. 59. Chemistry Manufacturing Control (CMC)CONFIDENTIAL 59
    60. 60. EP 94 – General properties  Tetrapeptide  Manufactured by solid phase peptide synthesis  Lyophilized powder  Solubility: Good solubility in water  Stability of drug substance:  Stable at +4°C for more than 2 years  Stability of drug product:  Stable at -18°C, +4°C and 25°C for more than 6 months in salineCONFIDENTIAL 60
    61. 61. Specification Test Method Specifications Appearance Visual observation Off-white to yellow powder Mass spectral analysis EP 2.2.43; ESI M = 571.2 +/- 1.0 Amino acid analysis Pre-column derivatisation Gly:1; Tyr:1; Dab:1; with OPA/Fmoc-Cl Phe (pNO2).1 EPA 2.2.56 RP-HPLC EP 2.2.29 Purity > 98.0% (area%) Related Impurities by EP 2.2.29 Sum impurities < 2% RP-HPLC Water content Karl Fischer, EP2.5.12 To be reported Acetate HPLC with UV detection To be reported Residual TFA HPLC with UV detection < 0.1% (tentative)CONFIDENTIAL 61
    62. 62. Specification cont’d Test Method Specifications Residual organic GC <USP 467> Individual according to ICH solvents Q3 Bioburden EP 2.6.12 Aerobic bacteria < 10 CFU/0.1g Anerobic bacteria < 10 CFU/0.1g <USP 61> Yeasts and moulds < 10 CFU/0.1g Bacterial endotoxins EP 2.6.14 <USP < 5 EU/mg (tentative) 85> Net peptide content Pre-column To be reported (NPC) derivatisation by AAA with OPA/Fmoc- Cl EPA 2.2.56 Mass balance Calculation To be reported NPC+acetate+w aterCONFIDENTIAL 62
    63. 63. EP 94  Manufactured by Polypeptide  Part no: SP080384  Storage temp: 4°CLot no HPLC analysis DateCF 07314 97.7% 26.3.2008CF 07314 95,8% 11.2.2010CONFIDENTIAL 63
    64. 64. EP 94  Manufactured by Polypeptide  Part no: SP080384B  Storage temp: 4°CLot no HPLC analysis DateMZ77114 98.9% 07.05.2009CONFIDENTIAL 64
    65. 65. Drug Product Stability of EP 94 in Saline Stability (non- ICH) of EP94, 0.1mg/mL in 9 mg/mL NaCl (preserved with NaN3 1mg/mL) Temp Time 0 14 days 2.5 mo 4.5 mo 6 mo -18°C 96.5 96.5 96.6 96.5 4°C 96.5* 96.5 96.6 96.4 96.2 Ambient 96.6 96.6 96.0 95.5 * Area %CONFIDENTIAL 65
    66. 66. CMC - T do list o  Drug Substance  Analytical development  Specification  Stability ICH  Manufacturing  Drug Product  Analytical development  Preformulation  Interaction with packaging mtr  Stability ICH  Manufacturing for tox and clinical  Regulatory  IND/CTA chapterCONFIDENTIAL 66
    67. 67. Refererences 1. Thom T, Haase N, et al. Writing Group Members. Heart Disease and Stroke Statistics--2006 Update. Circulation; A Report From the American Heart Association Statistics Committee and Stroke Statistics Subcommittee; January 11, 2006; 2006. CIRCULATIONAHA.105.171600. 2. Bishop E. Heart disease may actually be rising; researchers claim deaths are now being delayed to a later age group. Wall Street Journal. 1996 November 13, 1996; pB3(W) pB6(E) col 1 (11 col in) 3. Gerber Y, Jacobsen SJ, Killian J, Weston S, Roger VL. Impact of Participation Bias in a Population-Based Study of Myocardial Infarction in Olmsted County, Minnesota, 2002 to 2004. Circulation. 2006;13:e827. 4. Eribis Peptide 94 Reduces Infarct Size in Rat Hearts Via Activation of Centrally Located μ Opioid Receptors. J Cardiovasc Pharmacol. 2011 Nov 29 Gross GJ, Hsu A, Nithipatikom K, Bobrova I, Bissessar E. 5. Dose-dependent cardioprotection of enkephalin analogue Eribis peptide 94 and cardiac expression of opioid receptors in a porcine model of ischaemia and reperfusion. Eur J Pharmacol. 2012 Jan 15;674(2-3):378-83 Karlsson LO, Bergh N, Li L, Bissessar E, Bobrova I, Gross GJ, Akyürek LM, Grip L. 6. Opioid receptor agonist Eribis peptide 94 reduces infarct size in different porcine models for myocardial ischaemia and reperfusion. Eur J Pharmacol. 2011 Jan 25;651(1-3):146-51 Karlsson LO, Grip L, Bissessar E, Bobrova I, Gustafsson T, Kavianipour M, Odenstedt J, Wikström G, Gonon AT.CONFIDENTIAL 67

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