COPAXONE DEVELOPMENT AND STATE OF THE ART RUTH ARNON. The Weizmann Institute of Science Rehovot, ISRAEL

Transcription

1 COPAXONE DEVELOPMENT AND STATE OF THE ART RUTH ARNON The Weizmann Institute of Science Rehovot, ISRAEL

2 Research in academic institutions is mostly basic, namely curiosity-driven In some cases it reaches a stage when it becomes applicable Technology Transfer - leads to Applied Research Development by industry into a product Only a fraction of basic research is applicable and only a small fraction of it becomes applied

3 Two examples from my own research: 1. The development of Copaxone - a drug for the treatment of Multiple Sclerosis 2. Research towards the development of a broad-spectrum influenza vaccine

4 Multiple Sclerosis (MS) An inflammatory demyelinating disease of the CNS, characterized by multifocal plaques in MRI, with an autoimmune etiology. T-cells specific to one or various myelin antigens attack the myelin resulting in destruction of the myelin envelope, axonal loss, interference in nerve conduction and severe impairment. Relapsing/Remitting (RRMS) Progressive Relapsing Multiple Sclerosis (PRMS) Secondary Progressive (SPMS) Primary Progressive (PPMS)

5

6

7 Experimental Autoimmune Encephalomyelitis (EAE) - A Model of MS Can be induced in several species by several autoantigens such as MBP, PLP, MOG. Mediated by Th1-type CD4 T cells, reactive to myelin encephalitogenic epitopes. Perivascular infiltration of T-cells into the brain

8 Pre-existing Information CNS antigen - Myelin Basic Protein Proteins are antigenic - induce an immune response Synthetic, protein-like polymers, are also antigenic - synthetic antigens Conjugates of such polymers with lipids induce anti-lipid immune response

9 Working Hypothesis A basic synthetic polymer with positive electrical charge may mimic the Myelin Basic Protein It might form complexes in vivo with acidic lipid molecules Such a polymer could have encephalitogenic properties If so - it could provide a good research tool

10 COPAXONE (Copolymer 1) Also known as Glatiramer Acetate(GA) Synthetic copolymer of amino acids Designed to simulate Myelin Basic Protein Resembles MBP in its structure and immunological properties

11

12

13

14

15

16

17 TEVA undertakes development Upscaling and development of an industrial production process Validation of processes and analytical procedures Quality assurance Clinical Trials

18

19

20

21

22 Time Frame Basic research on animal models including primates (WIS) Applicable research including clinical trials (WIS and medical schools) Applied research and Development ( industry - TEVA ) Approval of Copaxone by the FDA

23 Yearly Relapse Rate While on GA 1.25 Mean Relapse Rate * Year # of pts : *ARR in the 2 years before GA start After year 9, mitt data included only those of patients randomized to GA in the double-blind phase of the study. Crossovers from placebo group had not yet received GA

24 In parallel to the development of Copolymer 1 as a drug for MS treatment, basic research continued and studies were carried out in our laboratory to explore the mechanism of its action

25 TCR Th2/3 MHC antagonist inducer blocking T H 2 Encephalitogenic T H -cell specific to MBP Diverse signal T H 2 T H 2 GA T H 2 T H 2

26

27 C Cop 1 induced cells penetrate and accumulate in the brain - the organ in which the pathological processes of EAE and MS occur A B Brain sections from a mouse adoptively transferred with Hoechst labeled Cop 1-specific T cells, seven days after their injection to the periphery.

28 Immunohistochemical analysis of cytokine expression by GA-specific cells in the brain

29 The effect of GA-specific cells on IFN-γ expression in the brain EAE-induced mouse injected with GA-specific cells EAE-induced mouse not injected with GA-specific cells normal mouse Staining of corresponding regions in the cortex with IFN-γantibodies

30 The effect of GA on Th-17 and Tregs in the CNS of EAE-induced mice GA reduces Th-17 inflammation and induces regulatory T- cells in the CNS of mice with relapsingremitting or chronic EAE. Aharoni et al. J. of Neuroimmunol. 2010

31 Conclusion - GA induces immunomodulation specific Th2 cells GA-specific Th2 cells from the periphery, either actively induced by treatment, or passively by adoptive transfer, reach the CNS and exert their activity in situ. The functionality of GA-specific cells in the CNS can be possibly attributed to their cross-reactivity with MBP, since they might be stimulated in situ by the MBP. Once activated in the target organ, the GA-specific cells secrete anti inflammatory cytokines that suppress inflammatory responses to other antigens in their vicinity and induce bystander-suppression.

32 Does GA induce neuroprotection? Neuroprotection is an effect that is supposedly induced by neurotrophic factors and may result in salvage, recovery or regeneration of the nervous system, its cells, structure and function. BDNF (brain-derived neurotrophic factor) plays a major role in this function. Additional factors NT3 and NT4

33 BDNF Expression by GA-specific cells in the brain Activated labeled GA specific cells were injected into the peritoneum of either normal or EAE induced mice. After seven days mice were perfused and brains were stained immunocytochemically for BDNF expression.

34 GA treatment restores the impaired BDNF expression induced by the disease EAE induced mice with chronic disease (grade 3) were injected daily with GA. Two weeks after the first injection, BDNF expression was detected immunohistochemically and by in situ hybridization. * statistical significance from naïve mice # statistical significance from untreated EAE induced mice. The immunomodulator GA augments the expression of neurotrophic factors in brains of EAE mice. Aharoni, Eilam, Domev, Sela, Arnon PNAS, 2005.

35 GA augments the expression of the neurotrophic factors NT3 and NT4

36 Since MS and EAE are demyelinating diseases, does GA have any effect on demyelination?

37 Description of the Experimental Model Clinical manifestations of EAE induced by MOG peptide 35-55

38 The effect of GA on myelin visualized by immunohistochemistry

39 The effect of GA on myelin visualized by WETSEM

40

41

42 Conclusion Copaxone arrests demyelination, possibly induces remyelination as well Can Copaxone lead to repair of damaged tissue?

43 Description of the Experimental Model Clinical manifestations of EAE induced by MOG peptide YFP 2.2 transgenic mice are similarly susceptible to EAE as C57BL/6 mice GA ameliorated EAE regardless of the stage in which it was administered Neurogenesis and neuroprotection induced by periphral immunomodulatory treatment of EAE. Aharoni, Arnon, Eilam. J. Neuroscience, 2005.

44 The effect of GA on histological manifestations of MOG induced EAE

45 Penetration of neuronal progenitors into injury sites EAE + GA EAE untreated Cells expressing DCX migrating from the RMS towards a lesion in the striatum. Cells co-expressing BrdU and DCX inside a lesion in the frontal cortex, accompanied by axonal sprouting and extension of fibers into the lesion.

46 Pyramidal neuron in the cortex (occipital, layer 6), born during the concurrent injections of the proliferation marker BrdU and GA, to EAE-induced mice. One month after completion of treatment, neurons co-expressing BrdU (yellow) and YFP (green) with apical dendrites and axon are seen, indicative of mature functional neurons.

47 MECHANISM OF ACTION,! I. By induction of an immunomodulating effect on different levels of the immune response. II. By induction of neuroprotection and neurogeneration III. By interfering with the demyelination

48 CONCLUSIONS Basic research, curiosity-driven, may lead to application and has resulted in the development of Copaxone as a drug for Multiple Sclerosis Further support of the research enabled the deciphering of its mode of action, which is state of the art and could be valuable for future developments (e.g. other indications)

49 Acknowledgements Rina Aharoni Michael Sela Dvora Teitelbaum Raya Eilam Basak Kayhan Elizabeta Aizman Ariel Stock Anya Vainshtein Renana From Ido Form Vera Shinder TEVA

50 THANK YOU For your Attention

51

52 Development of a Peptide-Based Influenza Vaccine

53

54 Current vaccines Formalin inactivated whole virus Split virus vaccine (virus structure broken by ether, the soluble fraction is used for immunization) Subunit vaccine (the HA and NA are isolated from the membrane and complexed) Attenuated virus (cold adapted)

55 Our Approach - New concept Epitope-based vaccine Using small regions of the viral proteins that are conserved among many influenza strains Expression by genetic engineering Using a mixture of several such regions that will lead to maximal protection against many strains in broad population

56

57

58

59

60

61

62

63 Influenza virus - human epitopes Epitope HLA RESTRICTION HA (B epitope) SKAFSNCYPYNVPNYASL HA (Th epitope) PKYVKQNTLKLAT DR1, DR2, DR3, DR4, DR5, DR7, DR9, Dw13, Dw14 Dw52A. NP (CTL epitope) A2, A3, Aw68.1, B37. SAAFEDLRVLSFIRGY NP B8, B27. ELRSRYWAIRTRSG

64

65

66

67

68 Conclusions Peptide-based vaccines can elicit protection when the peptides are chemically attached to a carrier or expressed in it by recombinant methods. Such vaccines may be endowed with built-in adjuvanticity. Immunization with a mixture of peptides (or constructs expressing them) which represent B- and T-cell epitopes, confers long-term and cross-strain immunity against influenza. This approach could be adopted for the development of a cross-strain influenza vaccine for human use.

69 Research Achievements Selection of appropriate epitopes Effective protection of mice against infection Protection against a variety of influenza strains

70 BiondVax s Universal Vaccine Company Market Science Product Development Plan The Conserved Epitope Approach BiondVax s Universal Vaccine uses a unique and proprietary combination of selected parts of the viral proteins (epitopes) that are conserved and common to most influenza strains: and its advantages The selected parts of the viral proteins (peptides) used are: Immunogenic (i.e. epitopes) Conserved & Common Combination (B-Cells & T-Cells) resulting in: Selected Epitopes from HA, NP, M proteins One formulation Broad range efficacy against all strains tested Including pandemic flu (H5N1) No hen-egg allergy Year-round vaccination Short production cycle Long-term storage A Single, Universal Formulation: ONE VACCINE: For ALL flu strains, seasonal & pandemic

71 Phase I/II Clinical Trials Company Market Science Product Development Plan First Clinical Trial Young Adults Trial Design Phase I/II ( 18-49): started in Q2 (June); completed December 2009 Randomized, Single-Blind, Placebo-controlled, Escalating Double-Dose Safety Study of an Intramuscular Influenza Vaccine (Multimeric-001) Injected to 63 Healthy Volunteers Primary endpoint: Safety Secondary endpoint: Evaluation of immunological responses Results Safety: No significant side effects were observed Immunogenicity: Antibodies: Proliferation of antibodies against the vaccine and of antibodies that recognize the swine flu strain (A/H1N1) Antibody-mediated lysis by Complement Cells: T-Cell proliferation and cytokines secretion (IL-2, IFN-gamma)

72 Subsequent trials 1. Phase I/II in the elderly 2. Phase II in young adults 3. Phase II in the elderly (>65) All three trials were successful

73 Competitive Advantages Company Market Science Product Development Plan Parameter BiondVax Competitors Strain Multi-strain (incl. seasonal, Swine, Avian Flu) Strain-dependent Season Multi-season One season Safety Safe; No Allergy Hen egg allergy Production & Marketing Short production cycle (6-8 weeks) Annual, long production cycle (6-8 months) Vaccination All year round Limited to flu season

74 Time Frame Basic research in mice (WIS) Applicable research in humanized mice - proof of concept (WIS) Applied research in industry (BiondVax)

75 Acknowledgment: I. Copaxone Michael Sela Dvora Teitelbaum Oded Abramsky Murray Bornstein Rina Aharoni Masha Fridkis-Hareli Basak Kayhan II. Influenza Vaccine Michal Shapira Rafi Levi Tammy Ben-Yedidia Amiela Globerson Loya Abel Yair reisner Hadar Marcus

76 THANK YOU