Current Status of Gene Therapy Products in Japan. Teruhide YAMAGUCHI, Ph.D

Transcription

1 Current Status of Gene Therapy Products in Japan Teruhide YAMAGUCHI, Ph.D

2 Outline of the History of Gene Therapy in Japan Establishment of Expert Committee on Gene Therapy Guidelines for Gene Therapy Clinical Research (Health Science Council Report, MHW) Guidelines for Gene Therapy Clinical Research MHW Notice No.23) First gene therapy protocol was proposed for approval Start of gene therapy clinical research for ADA Deficiency Guidelines for Assuring the Quality and Safety of the Gene Therapy Products (MHW PAB Notice No.1062) Start of gene therapy clinical research for renal cell cancer... ~ Total of submitted clinical research protocols in Japan : 16 (approved :12) Amendments of Guidelines and Evaluation System on Gene Therapy Clinical Research

3 Evaluation of Clinical Study of Gene Therapy in Japan Clinical Research Head of the Organization Clinical Trial under the Regulation by Pharmaceutical Law Sponsor Report Submission of Protocol Report Submission of Protocol Prime Minister of MHLW Prime Minister of MHLW Judgment of newness Report Consultation YES No Report Consultation Report within one month Health Science Council Food and Pharmaceutical Affairs Council Guidelines for Gene Therapy Clinical Research Guidelines for Assuring the Quality and Safety of Gene Therapy Products

4 Regulation of Gene Therapy Products and Its Approval Fundamental Research Study on Materials as a GeneTherapy Drugs Physicochemical, Biological and Structural Characterization Screening Test Preclinical Study Application of Clinical Trial IND to Prime Minister of MHLW Clinical Trial Phase I Phase II Phase III Guidelines for Quality and Safety of gene therapy products Pharmaceutical Affairs Law 80. GCP Evaluation of Pre-clinical and Clinical Studies Approval as a New Drug

5 Summary of the Guidelines for Gene Therapy Clinical Research (1) Chapter : General Rules Target diseases Serious genetic diseases or life-threatening diseases such as cancer and AIDS Diseases which seriously damage the physical function of the patients Confirmation of Quality Genes and related materials transferred to the patients should be manufactured in accordance with GMP Genetically modification of human germ cells (including fertilized ovum and embryo) is prohibited Effectiveness and safety of the research can be predicted based on sufficient scientific knowledge

6 Summary of the Guidelines for Gene Therapy Clinical Research (2) Chapter : Protection of the Human Rights of Research Subjects Informed consent by document Chapter : System of Research and Review Tasks of the researchers, director, institution head, IRB Chapter : Procedures for Conducting Clinical Research The director of the research should prepare a project protocol including, (1) The purpose (2) Theoretical basis for the selection of the disease (3) Genes involved and the methods of transferring genes (4) Non-clinical research findings currently available (5) Safety evaluation from non-clinical studies (6) Basis for the conclusion that the research is feasible (7) Plan (8) Suitability of institutions where the planned research will be conducted (9) Current situations of research related to the planned research (10) Professional records and list of publications of researchers

7 Guidelines for Assuring the Quality and Safety of Gene Therapy Products This guideline identifies the major issues concerning the assurance of quality and safety of the gene therapy products and outlines the data and information to be addressed by manufacturers when filing an application with respect to the quality and safety of gene therapy products intended for clinical use. Chapter 1 General provisions Chapter 2 Manufacturing process Chapter 3 Specifications and formulation Chapter 4 Stability Chapter 5 Preclinical safety studies Chapter 6 Tests for effectiveness Chapter 7 Pharmacokinetics and pharmacodynamics Chapter 8 Manufacturing facilities and equipment Chapter 9 Ethical consideration Chapter 10 Miscellaneous provisions

8 Chapter 1 General Provisions Purpose: To establish basic requirements for assuring quality and safety of gene therapy products Definition of gene therapy: To administer genes or genetically modified cells to the subject for treatment of diseases

9 Chapter 2 Manufacturing Process Clarification of the details of the manufacturing processes and their validation are critical components for the assurance of quality and safety of the gene therapy products Relevant information concerning the manufacturing process, which differs depending on the type of gene transfer and administration method, should be described Type of gene transfer» Viral vector» Non-viral vector» Direct transfer Administration method» Ex vivo» In vivo

10 Manufacturing Process - Viral Vectors - The reasons for selection of the viral vector and its specific features Biological features of the wild type virus and its harmful effects on humans Genetic construct - Construction, structural analysis and characterization of genetic construct - Structure and biological activity of expression product Other DNAs Culture methods and biological features of packaging cells and potential harmful effects of the packaging cells on humans Potential harmful effects of viral vector-producing cells on humans Features of particle structure of the viral vector Biological features of the viral vector Manufacturing method Cell banking system

11 Manufacturing Process - Non-Viral Vectors - Theoretical and experimental rationale for selection of non-viral vectors as gene transfer method Genetic construct Other DNAs The manufacturing procedures and purification methods Structure and composition of non-viral vectors Biological features Cell banking system

12 Manufacturing Process - Direct Transfer of DNA or RNA - How to manipulate the naked nucleic acid so as to introduce it into target cells or humans (The gene transfer procedure, reagent, apparatuses..) Biological features of the direct transfer method (Tissue specificity, efficiency ) The rationale for the gene transfer methods Genetic construct Cell banking system

13 Manufacturing Process - Classification by Administration Methods - The administration methods should be appropriately designed and justified Ex vivo methods Target cells Donor selection criteria Cell culture procedures Acceptance criteria of transduced cells Method of administration of transduced cells to the patients In vivo methods Target cells Administration method Possibility of transfer of genes into non-target cells (especially germ cell lines)

14 Chapter 3 Specifications and Formulation Validation of the production process and the quality control of the raw materials as well as the setting of specifications of the final products should be performed appropriately Purity tests for raw materials, contaminants, process-related impurities, product-related impurities Freedom from bacteria, adventitious viruses, mycoplasmas, fungi and endotoxin Appropriateness of formulations Procedures for lot-to-lot production control of bulk and final product Examples of specifications and test methods: Properties The suitable identification tests A limiting test for impurities and toxic contaminant denial test The potency or quantitative biological activity of expression products of transgenes or related products Cell viability in the case living cells should be included

15 Chapter 4 Stability Taking into consideration the distribution and usage period, appropriate stability tests should be performed, and the storage conditions and expiration period should be set on the bulk and final products The storage in other conditions prescribed or beyond the expiration period should be examined to define the limits of their stability The rationale for the appropriateness of the lot number used in each test should be provided When carrying out the stability tests, the relevant guidelines should be consulted where necessary and appropriate

16 Chapter 5 Preclinical Safety Studies The appropriate tests on the products using suitable animals and/or in vitro systems should be performed. The safety tests should reflect the routes of administration of the products in humans In particular, the following items should be considered Replication competent virus Cytotoxicity Genetic integration to chromosomes Abnormal expression of the transgenes Carcinogenicity Immunogenicity General preclinical toxicity tests, when appropriate and possible

17 Chapter 6 Tests for Effectiveness The efficiency of gene transfer Structure and stability of the transgenes The expression efficiency from transgenes and their stability Biological activities of the expression products Intended effects on cells, tissues and individuals etc. should be examined by appropriately designed tests using cultured cells and experimental animals When appropriate model animals for the intended disease are available, the therapeutic effects using them should be examined

18 Chapter 7 Pharmacokinetics and Pharmacodynamics It is necessary to estimate the life span of genetically modified cells in humans and explain that the intended effects can be accomplished sufficiently, through tests on the in vivo kinetics including absorption, distribution, etc. of gene therapy products or transduced cells in test animals Especially, in the case where gene therapy products or genetically modified cells are targeted to reach a specific site (tissues, etc.), their localization should be sufficiently explained In performing pharmacokinetic tests, refer to the relevant guidelines

19 Summary of Non-clinical Test Results Data from the manufacturing process to pharmacokinetics and pharmacodynamics described so far, should be summarized The safety of gene therapy products is adequately guaranteed based on current knowledge, and conducting the clinical studies is justified from the viewpoints of quality, safety, and expected efficacy, should be demonstrated

20 Outline of Clinical Studies Current knowledge about the diseases selected as indications Plan for the clinical study on gene therapy Justification of conducting gene therapy clinical research Patient selection and exclusion criteria Method of obtaining patient consent The number of patients required for clinical evaluation and the terms of clinical research, and their rationales Methods used in the gene therapy clinical research Schedule for follow-up of patients Potential for gene transfer to persons who are not patients

21 Current Status of Gene Therapy Clinical Research in Japan

22 Gene Therapy Experience Number of protocols Japan 16 USA 469* Europe 99* As of July, 2002 As of Sept.,2001 As of Sept.,2001 Reproduced with permission from the Wiley Journal of Gene Medicine web site

23 Year 1995 Institution Gene therapy protocols (1) Hokkaido Univ. Target ADA deficiency Vector Retro Gene ADA Pts/ Cases (Planed) Tokyo Univ. Renal Cell Carcinoma Retro GM-CSF Okayama Univ./ RPR Gencell Lung Cancer Adeno p53 8(24) # 2000 Chiba Univ./ RPR Gencell Esophageal Cancer Adeno p53 3(29) # 2000 Cancer Chemotherapy Center Breast Cancer Retro MDR1 2 #Company sponsored As of April,2002

24 Gene therapy protocols (2) Year Institution Target Vector Gene Pts/ cases (Planed) 2000 Nagoya Univ. Malignant Glioma Liposome IFN Jikei Univ. RPR Gencell Lung Cancer Adeno p53 1(24) # 2000 Tohoku Univ. RPR Gencell Lung Cancer Adeno p53 1(24) # 2000 Okayama Univ. Prostate Cancer Adeno HSV-TK Tokyo Medical Univ. RPR Gencell Lung Cancer Adeno p53 2 (24) # #Company sponsored As of April,2002

25 Gene therapy protocols (3) Year Institution 2000 Osaka Univ. Tsukuba Univ. Tokyo Univ. Jikei Univ. Hokkaido Univ. Tohoku Univ. Target PDA Buerger s Disease Leukemia Osteomyelo- dysplasia Neuroblastoma Colon carcinoma (liver metastasis) ADA deficiency X-SCID Vector Plasmid Retro Adeno Adeno Retro Retro Gene HSV-TK LNGFR IL-2 Lptn Cytosine deaminase ADAcDNA hil-2r 2Rγc chain Pts/ Cases (Planed) 7(22) HGF 7(22) (10) (6) (18) (2) (5) As of April,2002

26 Target Diseases Genetic disease 3 (19%) Vascular disease 1 (6%) 12 (75%)

27 Gene Delivery Systems Plasmid Retrovirus Liposome 1 (6%) 1 (6%) 6 (38%) Adenovirus 8 (50%)

28 Company-Sponsored Clinical Trials 5 (31%) Investigator Investigator s s Clinical Researches 11 (69%)

29 No. of Gene Therapy Protocols Notarized Year

30 Protocol Review Time (n=2) 19.5(n=2) Month (n=6) 5.3(n=3) Year

31 How to promote and move forward - Quality - Viral vector standards for Viral dose definitions Reference Standard Material for Adenovirus Type 5 Reference Material for other vectors

32 How to promote and move forward - Safety - Germ line Integration Viral Shedding & Monitoring Somatic Cell Integration into Hot spots Replication Competent Viruses

33 CPE method vs infectivity PCR Classical Cell culture/cpe method Ad vector RCA Infectivity PCR Infection HeLa cells 1 9 days Cell pellet 7days DNA extraction by glass beads 1~2days 4wks Cell lysate A549cells Quantification of E1 DNA by TaqMan PCR 14days 7days Cytopathic effect CPE) Long term Non-quantitative Sensitivity : 1 pfu in starting material Rapid Sensitive Quantitative

34 S+L- assay method vs. Infectivity PCR <Classical extended S+L- assay method> Retrovirus vector RCR Infection Mus dunni cells <Infectivity qrt-pcr with RCR concentration> 3-10 days Culture sup 4wks 3,4 days Passage x 5 Concentration of RCR by PEI-magnetic beads 1 day Infection Culture sup PG-4(S+L-) cells DNA extraction from PEI-beads fraction 7 days Focus formation Long term Non-quantitative Quantification of Env RNA by TaqMan qrt-pcr Sensitivity : 1ffu in starting material Rapid High sensitivity Quantitative

35 How to promote and move forward - Efficacy - Development of More Effective Vectors Development of Vector to deliver the Gene of Interest to Target Cell/tissue/site with a High Efficiency Development of Vector to Express the Gene Efficiently and Stably in the Cell or Tissue

36 Special thanks for giving a lot of information on gene therapy in Japan to NIHS: Dr. Takao Hayakawa Dr. Eriko Uchida JPMA: Wataru Toriumi Hitoshi Kotani