APPENDIX III

FORMAT FOR SUBMISSION OF PRECLINICAL AND CLINICAL DATA*

FOR r-DNA BASED VACCINES, DIAGNOSTICS AND OTHER BIOLOGICALS

(Reproduced from Guidelines for Generating Preclinical and Clinical Data for r-DNA based vaccines, diagnostics and other biologicals issued by Department of Biotechnology, Ministry of Science and Technology, Govt. of India)

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*For details to generate these data, please consult the document entitled “ Guidelines for generating preclinical and clinical data for r-DNA based vaccines, diagnostics and other biologicals”.­­­­­­­­­­

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A :  SPECIFICATION AND CHARACTERIZATION INFORMATION ON r-DNA VACCINES AND BIOLOGICAL PRODUCTS

1. Description in details of the method of r-DNA products:

(a)  host cells,

(b) gene construct,

(c) vector construction including a description of the source and function of the component parts of the vectors,

(d) source and diagram of the plasmid(s) used,

(e) all intermediate cloning procedures, and

(f)  transfection methods.

2.      Description of the method of sequence verification (such as restriction enzyme mapping, PCR etc.).

3     Description on Identity-Physical, Chemical, Immunological and Biological wherever applicable

(a)  Description on recombinant DNA products :

(1)   Primary structure (Amino acid sequences)

(2)   Secondary structure (disulfide linkages etc.)

(3)   Post-translation modification (glycosylation etc.)

(b)  Monoclonal antibodies (if applicable) :

-         identity by rigorous immunochemical and physicochemical characterization.

  4. Potency.

(a)  Production of specific antigen in transfected cell line,

(b)  Immune response in mice,

(c)   Hypersensitivity (Guinea pig maximization test), and

(d)  Permissible limits of potency.

5. General Safety Test.

6. Data on sterility tests as per Indian Pharmacopia guidelines.

 7. Data on purity of recombinant product.

(a)  Limits of purity,

(b)  Characterization of minor impurities like RNA, protein and genomic DNA,

(c)   Permissible limits of moisture, if lyophilized, and

(d)  Pyrogenicity

8.Description of constituent materials like preservatives etc.

9. Data on stability of finished formulation as per IP (Indian pharmacopia) guidelines.

B : DATA ON PRECLINICAL TESTING

1. Biological activity/ pharmacodynamics in vitro and in appropriate animal  models.
2. Safety Pharmacology (Functional indices of toxicity).
3. Toxicology and pharmacokinetics (Absorption, Distribution, Metabolism, Excretion- ADME)
4. Immunogenicity/Immunotoxicity
5. Reproductive and developmental toxicity
6. Genotoxicity studies
7.   Carcinogenicity studies

C: RECOMBINANT IMMUNODIAGNOSTIC REAGENTS

1. Specification and characterization of r-DNA diagnostic products (Please provide information as per     column1-9 under Section A of this format).
2. The data on the sensitivity / specificity / predictive positive value/ predictive negative value / overall    diagnostic accuracy of recombinant product in diagnostic assay.
3. Data on (1) “in-house” validation and (2) independent validation.
4.   Data using indigenous / internationally available panel of sera / clinical materials.

D: CLINICAL TRIALS

1. Phase I : Human/Clinical Pharmacology Immunogenic Potency

(a)  Details on level of specific antibodies including its kinetics in healthy subjects.

(b)  Details on cytokine profiles in healthy subjects.

(c)   Details on T-cell responses in healthy subjects.

(d)  Data on auto-antibodies and immune complexes in healthy subjects.

(e)  Details on haematological and clinical chemistry.

2. Phase II: Exploratory Clinical Trials- Preventive/Therapeutic Efficacy (Data to be generated in subjects residing in endemic/ non-endemic areas)

(a)  Protective / therapeutic potentials of r-DNA vaccines.

(b)  Details of the haematological data.

(c)   Details on the clinical chemistry.

(d)  Data on experiments on minimum protective / therapeutic dose vis-à-vis immune response (both T&B cells).

3. Phase III: Confirmatory Trials

(a)  Preventive / therapeutic effects.

(b)  Immunological / clinical chemistry parameters in some subjects belonging to different ethnic and socio-economic groups.