Files CDSCO DBTSimilarBiologicsfinal

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8.3 Con rmatory Safety and E cacy Study 20

8.4 Safety and Immunogenicity Data 21

8.5 Extrapola on of E cacy and Safety Data to other Indica ons 22

9. Data Requirements for Market Authoriza on Applica on 22

10. Post-Market Data for Similar Biologics 23

10.1 Pharmacovigilance Plan 23

10.2 Adverse Drug Reac on (ADR) Repor ng 23

10.3 Post Marke ng Studies (Pms) 23

11. Applica on Forms 25

12. Archiving of Data 25

13. Glossary 26

14. References 29

Annexes:

1: Protocols on Regulatory Pathway for Recombinant Pharma 30Products Adopted from Mashelkar Report

2: Requirement of Physicochemical and Biological Characteriza on of

2a: Nucleic Acid Based Recombinant Products 35

2b: Therapeu c Proteins 37

2c: Therapeu c Enzymes 39

2d: An bodies 41


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1. Introduc on The Guidelines on Similar Biologics prepared by Central Drugs Standard

Control Organiza on (CDSCO) and the Department of Biotechnology (DBT) laydown the regulatory pathway for a similar biologic claiming to be similar to analready authorized reference biologic .

The guidelines address the regulatory pathway regarding manufacturing processand quality aspects for similar biologics.

These guidelines also address the pre-market regulatory requirements includingcomparability exercise for quality, preclinical and clinical studies and post marketregulatory requirements for similar biologics.

2. Background & Objec vesThe CDSCO is the na onal regulatory authority in India that evaluates safety,e cacy and quality of drugs in the country. The DBT through Review Commi ee

on Gene c Manipula on (RCGM) is responsible for overseeing the developmentand preclinical evalua on of recombinant biologics.

Presently, several organiza ons are ac vely engaged in manufacturing andmarke ng similar biologics in India. So far, these similar biologics were approvedby RCGM and CDSCO using an abbreviated version of the pathway applicable tonew drugs on a case by case basis. Since there are several such products underdevelopment in India, both regulatory agencies considered the need to publisha clear regulatory pathway outlining the requirements to ensure comparable

Guidelines on Similar Biologics:Regulatory Requirements forMarke ng Authoriza on in India


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Guidelines on Similar Biologics: Regulatory Requirements for Marketing Authorization in India

safety, e cacy and quality of a similar biologic to an authorized reference

biologic. Based on demonstra on of similarity in the compara ve assessment, asimilar biologic may require reduced preclinical and clinical data package as partof submission for market authoriza on.

The objec ve of this document is to provide guidelines to applicants to enablethem to understand and comply with the regulatory requirements for theauthoriza on of similar biologics in India.

3. Applicable Regula ons And Guidelines The similar biologics are regulated as per the Drugs and Cosme cs Act, 1940, the

Drugs and Cosme cs Rules, 1945 (as amended from me to me) and Rules forthe manufacture, use, import, export and storage of hazardous microorganisms/gene cally engineered organisms or cells, 1989 (Rules, 1989) no ed underthe Environment (Protec on) Act, 1986. Various applicable guidelines are asfollows:

Recombinant DNA Safety Guidelines, 1990

Guidelines for genera ng preclinical and clinical data for rDNA vaccines,diagnos cs and other biologicals, 1999

CDSCO guidance for industry, 2008:

o Submission of Clinical Trial Applica on for Evalua ng Safety and E cacy

o Requirements for permission of New Drugs Approval

o Post approval changes in biological products: Quality, Safety and E cacy Documents

o Prepara on of the Quality Informa on for Drug Submission for NewDrug Approval: Biotechnological/Biological Products

Guidelines and Handbook for Ins tu onal Biosafety Commi ees (IBSCs), 2011


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4. Competent Authori es The competent authori es involved in the approval process are as follows:

Review Commi ee on Gene c Manipula on (RCGM) 1

RCGM func ons in the Department of Biotechnology (DBT), Ministry of Scienceand Technology, Government of India. In the context of similar biologics, RCGMis responsible for authorizing import/export for research and development andreview of data up to preclinical evalua on.

Gene c Engineering Appraisal Commi ee (GEAC)1

GEAC func ons under the Ministry of Environment and Forests (MoEF) asstatutory body for review and approval of ac vi es involving large scale use ofgene cally engineered organisms (also referred as living modi ed organisms)and products thereof in research and development, industrial produc on,environmental release and eld applica ons.

Central Drugs Standard Control Organiza on (CDSCO) 2

CDSCO, headed by the Drug Controller General of India (DCGI) is the apex regulatory body under Ministry of Health & Family Welfare (MoHFW),Government of India which is responsible for the approval of new drugs.In the context of similar biologics, CDSCO is responsible for grant of import/ export license, clinical trial approval and permission for marke ng and manufacturing. State Food and Drug Administra on (FDA) works withCDSCO in each state and is responsible for issuance of license to manufacturesimilar biologics in India.

1RCGM and GEAC are statutory commi ees set up as per provisions of Rules, 1989 2CDSCO func ons as per the provisions of the Drugs and Cosme cs Act, 1940


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5. ScopeThese guidelines apply to similar biologics that contain well characterized proteins astheir ac ve substance, derived through modern biotechnological methods such asuse of recombinant DNA technology. The demonstra on of similarity depends upondetailed and comprehensive product characteriza on, preclinical and clinical studiescarried out in comparison with a reference biologic.

Similar biologic can only be developed against an authorized reference biologicthat has been approved using a complete data package in India. In case thereference biologic is not authorized in India, it should have been licensed andmarketed for at least 4 years with signi cant safety and e cacy data. In caseof no medicine or only pallia ve therapy is available or in na onal healthcareemergency, this period of 4 years may be reduced or waived o .

Any product can be considered as similar biologic only if it is proven to besimilar using extensive quality characteriza on against the reference biologic.Further product development should only be considered once the similarity ofthe product / molecule is demonstrated in quality.

The guidelines are applicable for similar biologics developed in India or importedinto the country. Detailed regulatory pathways for indigenously developed and

imported products3

are given in Annexure 1.6. Principles for Development of Similar Biologics

Similar biologics are developed through sequen al process to demonstrate thesimilarity by extensive characteriza on studies revealing the molecular andquality a ributes with regard to the reference biologic.

Although the extent of tes ng of the similar biologic is likely to be less thanthat required for the reference biologic, it is essen al that the tes ng of thesimilar biologic be su cient to ensure that the product meets acceptable levels

of safety, e cacy and quality to ensure public health. Generally, a reduc on in data requirements is possible for preclinical and /

or clinical components of the development program by demonstra on ofcomparability of product (similarity to authorized reference biologic) andthe consistency in produc on process, which may vary depending on thecharacteris cs of the already authorized reference biologic.

3Adopted from Report of the Task Force on Recombinant Pharma , 2005, chaired by Dr. R.A. Mashelkar, DG, CSIR(commonly referred as Mashelkar Report)


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Iden ca on of any signi cant di erences in safety, e cacy and quality studies

would mean the need for a more extensive preclinical and clinical evalua onand the product will not qualify as a similar biologic.

In case the reference biologic is used for more than one indica on, thee cacy and safety of the similar biologic has to be jus ed and if necessarydemonstrated separately for each of the claimed indica ons. Jus ca on willdepend on clinical experience, available literature data and whether or not thesame mechanism of ac on is involved in speci c indica ons.

6.1 Selec on of Reference Biologic

Reference biologic which is authorized using complete dossier is cri cal for thedevelopment of similar biologic. The ra onale for the choice of the referencebiologic should be provided by the manufacturer of the similar biologic in thesubmissions to the DBT and CDSCO.

The reference biologic has to be used in all the comparability exercise withrespect to quality, preclinical and clinical considera ons. The following factorsshould be considered for selec on of the reference biologic:

The reference biologic should be licensed in India and should be innovatorproduct. The reference biologic should be licensed based on a full safety,

e cacy and quality data. Therefore another similar biologic cannot beconsidered as a choice for reference biologic. In case the reference biologic is not marketed in India, the reference biologic

should have been licensed and widely marketed for 4 years post approvalin innovator jurisdic on in a country with well established regulatoryframework. In case no medicine or only pallia ve therapy is available or inna onal healthcare emergency, this period of 4 years may be reduced orwaived o .

The same reference biologic should be used throughout the studiessuppor ng the safety, e cacy and quality of the product (i.e. in thedevelopment programme for the similar biologic)

The dosage form, strength and route of administra on of the similarbiologic should be the same as that of the reference biologic.

The ac ve substance (ac ve ingredient) of the reference biologic and thatof the similar biologic must be shown to be similar

The acceptance of an innovator product as a reference biologic for evalua on ofsimilar biologic does not imply approval for its use in India.


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6.2 Manufacturing Process

The manufacturing process for similar biologic should be highly consistentand robust. If the host cell line used for the produc on of reference biologicis disclosed, it is desired to use the same cell line as the reference biologic.Alterna vely any cell line that is adequately characterized and appropriatefor intended use can be used to develop a similar biologic, with appropriate jus ca on in order to minimize the poten al for signi cant changes incri cal quality a ributes of the product and to avoid introduc on of certaintypes of process related impuri es that could impact clinical outcomes andimmunogenicity. For the establishment and characteriza on of the cell banks,the guidelines issued by the Interna onal Conference on Harmonisa on ofTechnical Requirements for Registra on of Pharmaceu cals for Human Use(referred to as ICH) viz. Q5A 4, Q5B5 and Q5D 6 should be referred for guidance.

The data requirements for review of manufacturing process at preclinicalsubmission stage include a complete descrip on of the manufacturing processfrom development and characteriza on of cell banks, stability of clone, cellculture/ fermenta on, harvest, excipients, formula on, puri ca on, primarypackaging interac ons (if di erent from reference biologic), etc. and theconsequences on product characteris cs as indicated below:

6.2.1 Molecular Biology Considera ons

The details regarding host cell cultures (including viral clearance), vectors, genesequences, promoters etc. used in the produc on of similar biologics shouldbe provided with appropriate drawings/ gures. The details of post-transla onalmodi ca ons if any (glycosyla on, oxida on, deamida on, phosphoryla onetc.) should be explained.

4ICH Q5A(R1): Viral Safety Evalua on of Biotechnology Products Derived from Cell Lines of Human or Animal Origin5ICH Q5B: Quality of Biotechnological Products: Analysis of The Expression Construct In Cells Used for Produc on ofR-DNA Derived Protein Products6ICH Q5D: Deriva on and Characteriza on of Cell Substrates used for Produc on of Biotechnological/Biological Products


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6.2.2 Fermenta on Process Development

At least three batches of reproducible fermenta on data at pilot scale(batch size adequate to give enough puri ed product to generate preclinicaldata).

Fermenta on process should be carried out in controlled and monitoredenvironment.

Details of fermenta on kine cs data from a representa ve batch indica ngcell growth, product forma on, pH, temperature, dissolved oxygen, majornutrient consump on pa ern and agita on rate.

Concentra on to be de ned in terms of product/litre, yield and volumetricproduc vity.

Data to verify that the speci c protein yield (amount of protein per unitcell mass) remains constant for all fermenta on batches.

Demonstrate that the overall produc vity is reproducible and scalable.

6.2.3 Downstream Process Development

Steps involved in puri ca on of protein. Batch size for protein puri ca on.

Descrip on of each unit opera on step during puri ca on and recovery ofprotein along with quan ta ve recovery of product at each stage.

Describe the quality of the refolded protein if the star ng material isaggregated or from inclusion bodies and include details of the refoldingprocess, speci c ac vity at di erent doses, dose response curve, stabilitydata and con rma on of solubility and absence of aggrega on.

Consistency of recovery in 3 consecu ve batches of puri ca on from 3independent batches of fermenta on


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For clinical trial applica on, addi onal requirements are applicable as per CDSCO

guidelines. A well-de ned manufacturing process with its associated processcontrols assure that an acceptable product is produced on a consistent basisin accordance with Good Manufacturing Prac ce (GMP). Data for submissionshould include:

Detailed descrip on of the drug substance and drug product processes

Cri cal quality a ributes of the product

Manufacturing process controls

Cri cal process parameters

Stability data Comparability of product manufactured at clinical scale against reference

biologic

Data from consistency batches and/or process valida on batches asapplicable

6.3 Quality Based Considera ons for Similar Biologics

6.3.1 Analy cal Methods The analy cal methods should be chosen for establishing product comparability

as per the cri cal quality a ributes of the product. For certain a ributes (e.g.product aggrega on) it is customary to use mul ple, orthogonal methods forcharacteriza on. Extensive state of the art analy cal methods should be appliedto detect even slight di erences in all relevant quality a ributes. IndianPharmacopoeia monograph should be followed, if available.

The measurement of quality a ributes in characteriza on should entail the

use of appropriately quali ed assays, which are reproducible and reliable. Themethods used to measure quality a ributes for batch release, stability studiesand in-process controls should be validated in accordance with ICH guidelines(ICH Q27, Q5C8, Q6B9), as appropriate.

7ICH Q2 (R1): Valida on of Analy cal Procedures: Text and Methodology8ICH Q5C: Stability Tes ng of Biotechnological/Biological Products9ICH Q6B: Speci ca ons: Test Procedures and Acceptance Criteria for Biotechnological/Biological Products


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The characteriza on studies should include samples of the applicants

recombinant product, reference biologic as control, known posi ve standardand nega ve control, wherever relevant. To ensure the sta s cal analysis, eachquan ta ve experiment should be done at least 3 mes and data should berepresented in terms of mean and standard devia on. Appropriate sta s calsigni cance should be represented throughout the characteriza on data.Physicochemical and biological characteriza on methods to be used forvarious categories of products viz. recombinant proteins, therapeu c enzyme,monoclonal an bodies etc. are given in Annexure 2 (2A-2D). It may be notedthat these Annexures are sugges ve but not limited to the speci ed methodand the requirements may vary on case by case.

6.3.2 Product Characteriza on

Characteriza on studies for similar biologics include physicochemical proper es,biological ac vity, immunological proper es, func onal assays, purity (process-and product-related impuri es etc.), contamina on, strength, and content.Principles outlined in the ICH Q6B guideline should be followed. IndianPharmacopoeia Monograph should be followed, if available.

i. Structural and Physicochemical Proper es: The analysis of physicochemicalcharacteris c should include determina on of primary and higher orderstructure of the product along with other signi cant physicochemicalproper es. The target amino acid sequence of the similar biologic shouldbe con rmed and is expected to be the same as for the reference biologic.Analy cal methods that are used (including biological and func onalassays) should have acceptable precision and accuracy. In cases, wherepost transla onal modi ca ons are taking place, these modi ca ons needto be iden ed and quan ed.

In case any signi cant di erences are found, these should be scien cally jus ed and cri cally examined in preclinical studies and clinical trials.

ii. Biological Ac vity: Biological products may have mul ple biologicalac vi es. In such cases, appropriate biological assays will be required tocharacterize the ac vity and establish the products mechanism of ac on


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and clinical e ects (in units of ac vity). The data from biological assays

will supplement the physicochemical characteriza on of the product asdescribed in the sec on 6.3.1.

Assays should be calibrated against an interna onal or na onal referencestandard, where available and appropriate. If no such standards areavailable, an internal reference standard must be established as per theICH guidelines. If the methods of bioassay(s) are documented in thespeci ca on, test(s) can be conducted accordingly.

iii. Immunological Proper es: The manufacturing process of recombinantbiologics is known to a ect the level of process related impuri es and posttransla onal modi ca ons of the product. These characteris cs may a ectthe immunogenicity of the product. Hence evalua on by characteriza on(an body or an body-derived product); comparison to reference biologicwith respect to speci city, a nity, binding strength and Fc func on; andevalua on by animal studies should be performed.

iv. Purity and Impuri es: Characteriza on of similar biologic requiresevalua on of the following via a combina on of analy cal procedures:

Product related variants (e.g., glycoforms, isomers etc.)

Product related impuri es (e.g., aggregated, oxidized or deamidatedproduct)

Host cell related impuri es (e.g., host cell protein, host cell DNA etc.)

Process related impuri es (residual media components, resin leachatesetc.)

Di erences observed in the purity and impurity pro les of the similarbiologic rela ve to the reference biologic should be evaluated to assesstheir poten al impact on safety and e cacy. Where the similar biologicexhibits di erent impuri es, those impuri es should be iden ed andcharacterized when possible. Depending on type and amount of theimpurity, conduct of preclinical and clinical studies will help to con rm thatthere is no adverse impact on safety and e cacy of the similar biologic.


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6.3.3 Speci ca ons

Speci ca ons of similar biologics are established around cri cal quality a ributesof the product with the intent of ensuring consistency in product quality andcomparability to reference biologic. Methods used for se ng speci ca ons mayor may not be same as the analy cal methods used for product characteriza onand for establishing product comparability. Acceptance limits should be setbased on reference biologic data and data from su cient number of batchesfrom preclinical or clinical batches.

6.3.4 Stability To set a shelf-life and storage condi on of drug product and drug substance, its

real me stability test should be conducted. Stability studies on drug substanceand drug product should be carried out using containers and condi ons thatare representa ve of the actual storage containers and condi ons, according torelevant guidelines (e.g. ICH Q5C 10 , WHO TRS 82211 ).

Side-by-side accelerated and stressed studies comparing the similar biologicto the reference biologic will be of value in determining the similarity of theproducts by showing comparable degrada on pro les.

6.4 Quality Comparability Study

The quality comparison between similar biologic and reference biologicis essen al. The applicant should submit a full quality dossier as per CDSCOguidance for industry, 2008 including the results of comparability exercise forthe similar biologic with the reference biologic before the applicant proposesto take the similar biologic to clinical development. First three consecu vestandardized batches which have been used to demonstrate consistency of themanufacturing process should be used.

Head-to-head characteriza on studies are required to compare the similarbiologic and the reference biologic at both levels of drug substance and drug

10 ICH Q5C: Stability Tes ng of Biotechnological/Biological Products11 Good manufacturing prac ces for biological products. In: WHO Expert Commi ee on Biological Standardiza on, Forty-second report, Geneva, World Health Organiza on, 1992, Annex 1 (WHO Technical Report Series, No. 822).


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product. In case the isola on of the drug substance is not possible, comparability

can be demonstrated at the drug product level with appropriate scien c jus ca on. Di erences between the similar biologic and the reference biologicshould be evaluated for their poten al impact on safety and e cacy of thesimilar biologic and addi onal characteriza on studies may be necessary.

Minor di erences between similar biologic and reference biologic in each qu alitycomponent can be there. Appropriate data should be submi ed to verify thatthese di erences do not impact on the safety and e cacy.

The quality comparison between the similar biologic and the reference biologic

should employ state-of-the-art analy cal techniques, including the analy calmethods that are sensi ve enough to detect the possibili es of changesto the product. The list of rou ne analy cal tests to be included for qualitycomparability exercise is given in Annexure-2 (2A-2D).

7. Data Requirements for Preclinical Studies

7.1 Prerequisite before Conduc ng Preclinical Studies

The applicant has to comply with the RCGM requirements like demonstra on ofconsistency of the process and product, product characteriza on and productspeci ca ons. The applicant should submit the data generated along with thefollowing basic clinical informa on and preclinical study protocols to RCGMfor obtaining permission. The toxicology studies should be ini ated a er theapproval of RCGM. The basic informa on about the reference biologic andsimilar biologic may include the following:

Basic informa on about the reference biologic

Informa on about the drug, route of administra on, absorp on andelimina on rate, therapeu c index, dose, vehicle, mode of administra on,dose response etc.

Bioequivalence range, if available.

Tissue-speci c localiza on , if available.

Available toxicity data on reference biologic.

Mode of ac on.


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Basic informa on about the similar biologic

Known / proposed clinical use

Target popula on (Age, sex, pregnancy, lacta ng, children etc.)

Dosage (frequency and intervals) units

Route / alternate routes of administra on

Final formula on + adjuvants, addi ves etc. - Toxicology data of adjuvants

Diluents

Presenta on e.g. pre lled syringe

The applica on to RCGM should be accompanied by approval by theIns tu onal Biosafety Commi ee (IBSC) of the applicant (copy of theminutes should be submi ed), and approval of Ins tu onal AnimalEthics Commi ee (IAEC), if available. The applicant should also providedetails of the proposed site for conduct of toxicity tes ng and personnelto be involved e.g. study director, principal inves gator, pathologist,other Inves gators and quality assurance o cer at the site. Status ofGLP cer ca on of proposed facility should also be provided.

7.2 Preclinical Studies (Pharmacodynamic and Toxicology Studies)

The preclinical studies should be conducted prior to the ini a on of any clinicalstudies. These preclinical studies should be compara ve in nature and designedto detect di erences if any, between the similar biologic and reference biologic.The preclinical study design may vary depending upon the clinical parameterssuch as therapeu c index, the type and number of indica ons applied.The approach adopted should be fully jus ed in the preclinical overview.Preclinical studies should be conducted with the nal formula on of thesimilar biologic intended for clinical use and for the reference biologic unlessotherwise jus ed. The dosage form, strength and route of administra on ofthe similar biologic should be the same as that of the reference biologic and incase of any di erences in these parameters, it should be jus ed.

The following studies are required for preclinical evalua on:


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7.2.1 Pharmacodynamic Studies

i. In vitro studies: Comparability of test and reference biologic should beestablished by in vitro cell based bioassay (e.g. cell prolifera on assays orreceptor binding assays).

ii. In vivo studies: In vivo evalua on of biological/ pharmacodynamic ac vitymay be dispensable if in vitro assays are available, which are known toreliably re ect the clinically relevant pharmacodynamic ac vity of thereference biologic. In cases where the in-vitro assays do not re ect thepharmacodynamics, In vivo studies should be performed.

7.2.2 Toxicological Studies

In case of in vivo toxicity studies, at least one repeat dose toxicity study in arelevant species is required to be conducted. The dura on of the study wouldbe generally not less than 28 days with 14 days recovery period. However thedura on may vary depending on the dosage and other parameters on case bycase basis.

Regarding the animal models to be used, the applicant should provide thescien c jus ca on for the choice of animal model(s) based on the dataavailable in scien c literature. However if the relevant animal species is notavailable and has been appropriately jus ed, the toxicity studies need to beundertaken in two species i.e. one rodent and other non rodent species, as perthe requirements of Schedule Y 12 with due permission from the RCGM.

Regarding the route of administra on, in cases when the relevant animal modelis used, the route of administra on would include only the intended route,whereas in all other cases, Schedule Y should be followed.

The dose should be calculated based on the therapeu c dose of the reference

biologic. If required a pilot dose response study should be conducted prior toini a ng the toxicity studies. Generally there would be three levels of doses(viz. low, medium and high) used in the animal toxicology studies correspondingto 1X, 2X and 5X of human equivalent dose or higher test dose for repeat dosetoxicity studies. Any di erence in the levels of doses should be jus ed and

12 Schedule Y: Requirements and Guidelines for Permission to Import and / or Manufacture of New Drugs for Sale or toUndertake Clinical Trials no ed as per G.S.R. 32(E), dt. 20.01.2005 under the Drugs and Cosme cs Rules, 1945


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approved prior to the studies. Regarding the schedule of administra on, the

therapeu c schedules may be used as the basis. Depending on the route of administra on, local tolerance should be evaluated.

If feasible, this evalua on may be performed as a part of above men onedrepeat dose toxicity study.

Accordingly the study groups of animals in repeat dose toxicity tes ng will consist of :

i. Historical Control (Op onal)

ii. Vehicle Control

iii. Vehicle Control for recovery group

iv. Formula on without protein (for vaccines) if mul ple adjuvants - each tobe checked independently

v. 1X similar biologic for study dura on ( lowest dose)

vi. 1X Reference biologic for study dura on

vii. 2X Medium dose similar biologic

viii. 5X High dose similar biologic

ix. Similar biologic with a recovery group going beyond the end of studyperiod for 7 to 14 days

The protocols and the study reports should provide complete details of varioussteps in the toxicity tes ng as indicated below:

Procedures prior to euthanasia e.g. blood drawing, body weight, etc.

Events immediately a er euthanasia, necropsy, gross descrip on, organweights and organs sampled for histopathology.

Biochemical parameters Equipment and methods used - units ofmeasurement and expression.

Haematology procedures and parameters method to be used (automatedor manual).

Sta s cal methods used.

Bone marrow either examined as an aspirate /smear or on histopathologysec on.


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In case of histopathological observa ons, the applicants should consider the

following points: Every observa on considered as devia on from described normal histology

needs to be documented and the incidence of each of these in the di erentgroups should be denoted

Whether such a feature is signi cant or not can be decided on reviewof sta s cal signi cance or dose response or if it is within or outsidethe normal range of values in case of biochemical and haematologicalobserva ons.

If all organs from all animals were not examined e.g. in 5 animals only 4livers were examined, the reason for the 1 liver not being examined shouldbe documented.

In case of premature death or morbidity the proposed course of ac on isto be included in the protocol.

Other toxicity studies, including safety pharmacology, reproduc ve toxicity,mutagenicity and carcinogenicity studies are not generally required forevalua on of a similar biologic unless warranted by the results from the repeatdose toxicological studies.

The nal report of the study should re ect all the aspects approved in theprotocol and the following addi onal sec ons/documents:

RCGM approval of protocol and test center

IBSC approval of report

IAEC approval for animal use and for the procedures QA statement

Signatures of study director and all inves gators who were involved in thestudy

All quality analy cal reports on the test material and vehicle

Animal feed and animal health cer ca ons


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Protocol devia ons if any

Discussion on the results

Individual animal data, summary data and any other data like computeranalysis outputs etc

Conclusion

7.3 Immune Responses in Animals

An body response to the similar biologic should be compared to that generatedby the reference biologic in suitable animal model. The test serum samplesshould be tested for reac on to host cell proteins.

For evalua ng immune toxicity of the similar biologic under study, the results oflocal tolerance (part of repeat dose or stand alone test) should be analyzed withthe observa ons regarding immunogenicity in sub-chronic study. Therefore, theimmunogenicity tes ng should be included as part of the sub-chronic repeatdose study while developing the protocols.

The other parameters for evalua ng immune toxicity include immune complexesin targeted ssues may be considered while evalua ng histopathologyobserva ons, etc.

A er comple on of preclinical studies the reports are submi ed to RCGM forreview and considera on.

Based on the successful evalua on of preclinical study reports includingdemonstra on of consistency of the process and product, productcharacteriza on, product speci ca ons and similarity to reference biologic,RCGM will recommend the applicant to approach DCG(I) to conduct appropriatephase of clinical trial as per the CDSCO requirements.

8. Data Requirements for Clinical Trial Applica on Besides the informa on submi ed in the preclinical applica on, the applicant

has to submit applica on for conduct of clinical trial as per the CDSCO guidancefor industry, 2008. The quality data submi ed should establish comparability ofsimilar biologic manufactured at clinical scale against reference biologic.


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8.1 Pharmacokine c Studies

Compara ve pharmacokine c (PK) studies should be performed in healthyvolunteers or pa ents to demonstrate the similari es in pharmacokine ccharacteris cs between similar biologic and reference biologic on case to casebasis.

The design of compara ve pharmacokine c studies should take the followingfactors into considera on.

Half life

Linearity of PK parameters

Endogenous levels and diurnal varia ons of similar biologic under study(where applicable)

Condi ons and diseases to be treated

Route(s) of administra on, and

Indica ons

Appropriate design considera ons can be combined into single dose or mul ple

dose studies with adequate jus ca on. These design considera ons include: Single dose, compara ve, PK studies

Parallel arm or

Cross over

Mul ple dose, compara ve parallel arm steady state PK studies

8.1.1 Single Dose Compara ve PK Studies

Dosage in the PK study should be within the therapeu c dose range of referencebiologic. Appropriate ra onale for dose selec on should be provided. The routeof administra on should be the one where the sensi vity to detect di erences isthe largest. Sample size should have sta s cal ra onale (i.e. sta s cally jus ed)and comparability limits should be de ned and jus ed prior to conduc ng thestudy.


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The analy cal method should be validated to have sa sfactory speci city,

sensi vity and a range of quali ca on with adequate accuracy and precision.It should have capability to detect and follow the me course of the similarbiologic (the parent molecule and / or degrada on products) in a complexbiological matrix that contains many other proteins.

Di erences in elimina on kine cs between similar biologic and reference biologic e.g. clearance and elimina on half life should be explored. Similarityin terms of absorp on / bioavailability should not be the only parameters of interest.

A parallel arm design is more appropriate for biologics with a long half life orfor proteins for which forma on of an bodies is likely or if study is being donein pa ents. In case of short half life, cross over design may be considered with ascien c jus ca on.

8.1.2 Mul ple Dose Compara ve PK Studies

Mul ple-dose, compara ve, parallel arm steady state PK studies are requiredfor a similar biologic that is used in a mul ple dose regimen, where markedly

higher or lower concentra ons are expected at steady state than that expectedfrom single dose data PK measurements, and where me-dependence anddose-dependence of PK parameters cannot be ruled out.

In case mul -dose compara ve PK studies are not done adequate jus ca onshould be provided.

8.2 Pharmacodynamic Studies

As for the PK studies in the similar biologic clinical development program, thepharmacodynamic (PD) studies should also be compara ve in nature.

Compara ve, parallel arm or cross-over, PD study in most relevant popula on(pa ents or healthy volunteers) is required for detec ng di erences betweenreference biologic and similar biologic. If PD marker is available in healthyvolunteers, PD in healthy volunteers can be done.

Compara ve PD studies are recommended when the PD proper es of thereference biologic are well characterized with at least one PD marker being linkedto the e cacy of the molecule. The rela onship between dose / exposure, the


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relevant PD marker(s) and response / e cacy of the reference biologic should

be well established and used to jus fy the design. The acceptance ranges forthe demonstra on of similarity in PD parameters should be prede ned andappropriately jus ed.

The parameters inves gated in PD studies should be clinically relevant and surrogate markers should be clinically validated. PD studies may be combined with PK studies, in which case the PK/PD rela onship should becharacterized.

PD study can also be a part of Phase III clinical trials wherever applicable.

8.3 Con rmatory Safety and E cacy Study

Informa on to establish compara ve safety and e cacy in relevant pa entpopula on is mandatory for all similar biologics.

Compara ve clinical trials are cri cal to demonstrate the similarity in safetyand e cacy pro les between the similar biologic and reference biologic withfew excep ons (e.g. recombinant human soluble insulin products for whichonly compara ve clinical safety study is required). The design of the studies

and the clinical comparability margins of the primary e cacy endpoints areimportant and should be given careful considera on and should be jus ed onclinical grounds. In line with the principle of similarity, equivalence trials withequivalence designs (requiring lower and upper comparability margins) arepreferred. If non-inferiority trials are required they must be clearly jus ed andapplicants are advised to consult with CDSCO prior to study ini a on. Samplesizes should have sta s cal ra onale and comparability limits should be de nedand jus ed prior to conduc ng the study.

The nature, severity and frequency of adverse events should be comparedbetween the similar biologic and reference biologic and should be based onsafety data from a su cient number of pa ents treated for an acceptable periodof me. E orts should be made to ensure that compara ve clinical studies havea su cient number of pa ents treated for acceptable period of me in order toallow detec on of signi cant di erences in safety between similar biologic andreference biologic.


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One or more adequately powered, randomized, parallel group, blinded

con rmatory clinical safety and e cacy trials are desirable based on thecomparability established during preclinical and PK / PD studies. More than onesafety and e cacy study may be required and the similar biologic will be treatedas a stand-alone product if the similar biologic is not comparable to referencebiologic in all preclinical evalua ons conducted and /or the PK/PD studies havenot demonstrated comparability.

The con rmatory clinical safety and e cacy study can be waived if all the belowmen oned condi ons are met:

i. Structural and func onal comparability of similar biologic and referencebiologic can be characterized to a high degree of con dence byphysicochemical and in vitro techniques

ii. The similar biologic is comparable to reference biologic in all preclinicalevalua ons conducted

iii. PK / PD study has demonstrated comparability and has preferen allybeen done in an in-pa ent se ng with safety measurement (includingimmunogenicity) for adequate period jus ed by the applicant and e cacy

measurements iv. A comprehensive post-marke ng risk management plan has been

presented that will gather addi onal safety data with a speci c emphasison gathering immunogenicity data

The con rmatory clinical safety and e cacy study cannot be waived if there isno reliable and validated PD marker.

8.4 Safety and Immunogenicity Data

Both pre-approval and post-approval assessment of safety is desired to beconducted for similar biologic.

Regarding pre-approval safety assessment, compara ve pre-approval safety dataincluding the immunogenicity data is required for all similar biologics includingthose for which con rmatory clinical trials have been waived. This pre-approvalsafety data is primarily intended to provide assurance of the absence of anyunexpected safety concerns.


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Compara ve safety data based on adequate pa ent exposure (both numbers

and me) must, in conjunc on with the published data on the referencebiologic provide assurance of absence of any unexpected safety concerns and inconjunc on with the proposed non-compara ve post-marke ng study providea comprehensive approach to the evalua on of safety of the similar biologic.

Post approval safety data requirements are elaborated in sec on 10.3.

8.5 Extrapolation of Efficacy and Safety Data to Other Indications

Extrapola on of the safety and e cacy data of a par cular clinical indica on(for which clinical studies has been done) of a similar biologic to other clinicalindica ons may be possible if following condi ons are met:

Similarity with respect to quality has been proven to reference biologic

Similarity with respect to preclinical assessment has been proven toreference biologic

Clinical safety and e cacy is proven in one indica on

Mechanism of ac on is same for other clinical indica ons

Involved receptor(s) are same for other clinical indica ons

New indica on not men oned by innovator will be covered by a separateapplica on.

9. Data Requirements for Market Authoriza on Applica on

The applicant should submit applica on for market authoriza on as perCDSCO guidance document for industry, 2008 . For cases where commercialmanufacturing is performed either at a di erent scale and/or with a di erentprocess as compared to that used for manufacturing phase III clinical trialbatches, then informa on on comparability of quality needs to be addi onallysubmi ed with appropriate jus ca on and will be dealt with on a case to case basis.


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10. Post-Market Data for Similar Biologics Though similar biologics are not new drug products and their risk will be similar

to reference biologic; however as similar biologics are authorized based on areduced preclinical and clinical data package, it is important to submit the RiskManagement Plan to monitor and detect both known inherent safety concernsand poten al unknown safety signals that may arise from the similar biologics.The reference biologic shall be maintained throughout the life cycle of theproduct.

The risk management plan should consist of the following:

10.1 Pharmacovigilance Plan

The clinical studies done on similar biologics prior to market authoriza on arelimited in nature so the rare adverse events are unlikely to be encountered. Hencea comprehensive pharmacovigilance plan should be prepared by manufacturerto further evaluate the clinical safety in all the approved indica ons in the post-marke ng phase. The pharmacovigilance plan should include the submission ofperiodic safety update reports (PSURs). The PSURs shall be submi ed every six

months for the rst two years a er approval of the similar biologic is grantedto the applicant. For subsequent two years the PSURs need to be submi edannually to DCGI o ce as per the Schedule Y.

10.2 Adverse Drug Reac on (ADR) Repor ng

All cases involving serious unexpected adverse reac ons must be reported tothe licensing authority within 15 days of ini al receipt of the informa on by theapplicant as per Schedule Y.

10.3 Post Marke ng Studies (PMS)

The clinical studies done on similar biologics prior to market authoriza on arelimited in nature so post marke ng studies should be conducted and the reportsbe submi ed to DCGI. The plan of post market studies should be captured inPharmacovigilance plan and update on the studies should be submi ed to theCDSCO.


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Regarding post-marke ng safety and immunogenicity study at least one

non-compara ve post-marke ng clinical study with focus on safety andimmunogenicity (on case by case basis) should be performed. This study mustbe designed to con rm that the similar biologic does not have any concernswith regards to the therapeu c consequences of unwanted immunogenicity.If immunogenicity is evaluated in clinical studies, it is not mandatory to carryout addi onal non-compara ve immunogenicity studies in post marke ngstudies.

The immunogenicity of the similar biologic should be evaluated usingappropriately designed studies with state-of-the-art methods, taking intoconsidera on the poten al impact on both safety and e cacy.

Ra onale on the strategy for tes ng immunogenicity should be provided. Assay methods should be validated and should be able to characterize an body content (concentra on or ter) as well as the type of an bodies formed.Of most concern are those an bodies that have poten ally serious impact onsafety and e cacy, such as neutralizing an bodies and an bodies with cross

reac vity. When neutralizing an bodies are detected in pa ents in clinicalstudies (either pre-approval clinical studies or post-approval clinical studies),the impact of the an bodies on the PK/PD parameters of the similar biologicshould be analyzed, where the data is available. Furthermore an assessmentof the impact of the neutralizing an bodies and cross-reac ng an bodies (ifapplicable) on the overall safety and e cacy of the similar biologic should beconducted.


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13. Glossary The de ni ons given below apply to the terms used in this guideline. They may

have di erent meanings in other contexts.

a. Comparability exercise: Comparison of a similar biologic with a referencebiologic with the goal to establish similarity in safety, e cacy and quality.

b. Drug: Drug includes (as de ned in Drugs and Cosme cs Act, 1940).

(i) all medicines for internal or external use of human beings or animalsand all substances intended to be used for or in the diagnosis,treatment, mi ga on or preven on of any disease or disorder inhuman beings or animals, including prepara ons applied on humanbody for the purpose of repelling insects like mosquitoes;

(ii) such substances (other than food) intended to a ect the structure orany func on of human body or intended to be used for the destruc onof (vermin) or insects which cause disease in human beings or animals,as may be speci ed from me to me by the Central Government byno ca on in the O cial Gaze e;

(iii) all substances intended for use as components of a drug includingempty gela ne capsules; and

(iv) such devices intended for internal or external use in the diagnosis,treatment, mi ga on or preven on of disease or disorder in humanbeings or animals, as may be speci ed from me to me by the CentralGovernment by no ca on in the O cial Gaze e, a er consulta onwith the Board.

c. Drug substance The ac ve pharmaceu cal ingredient and associated molecules that may be

subsequently formulated, with excipients, to produce the drug product. It maybe composed of the desired product, product-related substances, and product-and process-related impuri es. It may also contain other components such asbu ers.


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d. Drug product

A pharmaceu cal product type that contains a drug substance, generally inassocia on with excipients.

e. Equivalent

Similar or virtually iden cal in the parameter of interest. Equivalent e cacyof two medicinal products means they have similar (no be er and no worse)e cacy and any observed di erences are of no clinical relevance.

f. Gene c engineering

The technique by which heritable material, which does not usually occur or willnot occur naturally in the organism or cell concerned, generated outside theorganism or the cell is inserted into said cell or organism. It shall also mean theforma on of new combina ons of gene c material by incorpora on of a cellinto a host cell, where they occur naturally (self cloning) as well as modi ca onof an organism or in a cell by dele on and removal of parts of the heritablematerial (Rules, 1989).

g. Head-to-head comparison

Direct comparison of the proper es of the similar biologic with the referencebiologic in the same study.

h. Immunogenicity

The ability of a substance to trigger an immune response or reac on (e.g.,development of speci c an bodies, T cell response, allergic or anaphylac c

reac on).

i. Impurity

Any component present in the drug substance or drug product that is not thedesired product, a product-related substance, or excipient including bu ercomponents. It may be either process- or product-related.


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j. Manufacture

Manufacture in rela on to any drug includes any process or part of a processfor producing, altering, ornamen ng, nishing, packing, labelling, breaking upor otherwise trea ng or adop ng any drug with a view to its sale or distribu onbut does not include the compounding or dispensing in the ordinary course ofretail business; and to manufacture shall be construed accordingly.

k. Non-inferior

Not inferior to a comparator in the parameter studied. A non-inferiority clinicaltrial is one which has the primary objec ve of showing that the response tothe inves ga onal product is not clinically inferior to a comparator by a pre-speci ed margin.

l. Innovator product

A medicine which has been licensed by the na onal regulatory authori es onthe basis of a full registra on dossier; i.e., the approved indica on(s) for usewere granted on the basis of full safety, e cacy and quality data.

m. Pharmacovigilance

The science and ac vi es rela ng to the detec on, assessment, understandingand preven on of adverse e ects or any other drug related problems.

n. Reference Biologic

A reference biologic is used as the comparator for head-to-head comparabilitystudies with the similar biologic in order to show similarity in terms of safety,

e cacy and quality. Only a product that was licensed on the basis of a fullregistra on dossier can serve as reference biologic.

o. Similar

Absence of a relevant di erence in the parameter of interest.


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p. Similar biologic

A biological product/ drug produced by gene c engineering techniques andclaimed to be similar in terms of safety, e cacy and quality to a referencebiologic, which has been granted a marke ng authoriza on in India by DCGI onthe basis of a complete dossier, and with a history of safe use in India.

The products, where the reference biologic is not authorized in India shall beconsidered on a case by case basis if such products have been granted marke ngapproval in countries with well established regulatory systems such as US FDA,EMA etc. and have been in wider use for a minimum of four years.

Such products are also referred as biosimilars, similar biotherapeu c products,subsequent entry biologics or follow on biologics in various countries.

14. References i. EMEA guideline on similar biological medicinal products containing

biotechnology derived proteins as ac ve substance: non-clinical andclinical issues. London, 2006 (CHMP/BMWP/42832)

ii. EMEA guideline on immunogenicity assessment of biotechnology-derivedtherapeu c proteins London, 2007 (CHMP/BMWP/14327)

iii. ICH guideline on preclinical safety evalua on of biotechnology-derivedpharmaceu cals (S6), 1997

iv. Guideline for Safety Study of Biological Products, (KFDA, 2010)

v. World Health Organiza on (WHO) Guidelines on Evalua on of SimilarBiotherapeu c Products (SBP), 2009


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Annexure 1


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Nucleic acid based recombinantproducts - Physicochemical

Sequence (To prove if the sequencesame as reference biologic).

Restric on map for >1000 bp (To

check if secondary structure issame as reference biologic).

Purity on HPLC (To check if any impuri es are there).

Gel electrophoresis (agarose/acylamide/ urea page) (To checkquality of sample).

Southern/ Northern blot(Con rma on with referencebiologic).

Nucleic acid based recombinant products -Biological

Vector for expression of recombinant protein

Expression pa ern in actual target host cell

(To compare e ciency of expression ofsimilar biologic with reference biologic inthe target cell) .

Expression pa ern in closest animalspecies upon administra on (along withvehicle as nega ve control) (To comparee ciency of expression of similar biologicwith reference biologic in the target cellwhen administered in whole animal,this will evaluate the e ciency of vectorloca on and promoter ac vity in targetcell).

Kine cs of expression during the proposedtherapeu c period of protec on (Tocompare half life of the similar biologic

with reference biologic).

E cacy in appropriate disease/ infec onmodel in vitro and/or in vivo (To comparetherapeu c ac vity of the similar biologicwith reference biologic).

2A. Physicochemical and biological characteriza on of nucleic acidbased recombinant products

Annexure 2


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Absorp on spectrum from 190to 800 nm (To check similarity toreference biologic).

CD spectrum from 190 to 800 nm(To check secondary structuralchanges if any due to binding ofimpuri es).

Hybridiza on to the targetsequence. (To con rm withreference biologic).

Tm pro le (To check if anyimpuri es are present).

Es ma on of RNA and DNA usingnanodrop or reagent. (To checkconcentra on and impurity, if any

Absence of interference of markerenzyme/an bio c, if any (To comparetherapeu c interference and toxicity dueto a marker in the similar biologic withthat of reference biologic).

Vector for expression of siRNA/ snRNA etc.

Expression pa ern in actual target host cell(To compare e ciency of expression ofsimilar biologic with reference biologic inthe target cell)

Expression pa ern in closest animalspecies upon administra on (along withvehicle as nega ve control) (To comparee ciency of expression of similar biologicwith reference biologic in the target cellwhen administered in whole animal,this will evaluate the e ciency of vectorloca on and promoter ac vity in targetcell).

Kine cs of expression during the proposedtherapeu c period of protec on (Tocompare half life of the similar biologicwith reference biologic)

E cacy in appropriate disease/ infec onmodel in vitro and/or in vivo (To comparetherapeu c ac vity of the similar biologicwith reference biologic).

Absence of interference of markerenzyme/an bio c if any (To comparetherapeu c interference and toxicity dueto a marker in the similar biologic withthat of reference biologic).

Nucleic acid based recombinantproducts - Physicochemical

Nucleic acid based recombinant products -Physicochemical


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Therapeu c Proteins Physicochemical

Appearance, par culates,pH, osmolality, par cle size(if applicable) (To check

homogeneity).

MW, Sequence and amino acidcomposi on (To check purity).

N terminal sequence (atleast20 amino acid) (To check aminoacid sequence and structure).

Glycosyla on, Phosphoryla on,Acetyla on, and Myristoyla on,if any (To check if ac ve/inac ve form).

PEGyla on, esteri ca on,if applicable (To check ifmodi ca on is appropirate).

Tryp c map (1D and 2D) (Tocheck if secondary structure isconserved).

Sul ydryl groups(s) anddisulphide bridges (To checkif secondary structure isconserved).

Therapeu c Proteins Biological

Biological ac vity in actual target host cell(To compare ac vity of protein in similarbiologic with reference biologic in the

target cell).

Biological ac vity in closest animal species(if available) upon administra on (alongwith vehicle as nega ve control) (Tocompare ac vity of similar biologic withreference biologic in the target cell whenadministered in whole animal, this willevaluate the e ciency of vector loca on

and promoter ac vity in target cell). Kine cs of biological ac vity during the

proposed therapeu c period of protec on(To compare half life of the similar biologicwith reference biologic).

E cacy in appropriate disease/ infec onmodel in vitro and/or in vivo (If available)(To compare therapeu c interference and

toxicity due to a marker in the similarbiologic with that of reference biologic)

2B. Physicochemical and biological characteriza on of therapeu cproteins


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Size and Purity on HPLC (RP, SEC, IEX )/MALDI (To check if it is homogeneousand no impuri es are present ).

Isoform pa ern, if any (To check ifsecondary structure is conserved).

Gel electrophoresis (IEF, SDS PAGEand Na ve PAGE), Western blot (Toquali a ve check purity/ na vity).

Absorp on spectrum from 190 to800 nm (molar absop vity) (To checkpurity).

CD spectrum from 190 to 800 nm(To check if secondary structure isconserved)

Fluorescence spectrum (To check ifany impuri es such as quenchers arepresent).

FTIR spectrum, if applicable (To check if any prosthe c group ispresent).

NMR spectrum, if applicable (To checkif any prosthe c group is present).

A nity to the target receptor (Tocheck if required a nity to receptor isconserved).

Helix to Coil Transi on pro le (Toverify if the prepara on is stable andimpuri es or isofoms are a ec ng thestability).

Therapeu c Proteins Physicochemical


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Therapeu c Enzymes Physicochemical

Appearance, par culates,pH, osmolality, par cle size(if applicable) (To checkhomogeneity).

Sequence and amino acidcomposi on (To check purity).

Glycosyla on, phosphoryla on,acetyla on and myristoyla on,if any (To check if ac ve/inac ve form).

Pegyla on, estri ca on,if applicable (To check if

modi ca on is appropriate). Tryp c pep de map (1D and

2D) (To check if secondarystructure is conserved).

Size and purity on HPLC (RP,SEC, IEX)/ MALDI (To checkif secondary structure isconserved).

Gel electrophoresis (IEF,SDS PAGE and Na ve PAGE),Western blot (To qualita velycheck purity/ na vity).

Enzyme ac vity in gel assay inthe presence of chromogenicsubstrate (To check ac vity).

Therapeu c Enzymes Biological

Biological ac vity in actual target host cell(To compare ac vity of enzyme in similarbiologic with reference biologic in thetarget cell).

Biological ac vity in closest animal speciesupon administra on (along with vehicle asnega ve control) (To compare ac vity ofsimilar biologic with reference biologic inthe target cell when administered in wholeanimal, this will evaluate the e ciency ofvector loca on and promoter ac vity intarget cell).

Kine cs of biological ac vity during theproposed therapeu c period of protec on(To compare half life of the similar biologicwith reference biologic).

E cacy in appropriate disease/ infec onmodel in vitro and/or in vivo (To comparetherapeu c interference and toxicity dueto a marker in the similar biologic withthat of reference biologic).

2C. Physicochemical and biological characteriza on of therapeu cenzymes


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Absorp on spectrum from 190to 800 nm (To check purity)

CD spectrum from 190 to 800nm (To check if secondarystructure is conserved).

Helix to Coil Transi on pro le(To verify if the prepara on isstable and impuri es or isofomsare a ec ng the stability).

Fluorescence spectrum (Tocheck if any impuri es such asquenchers are present).

Km with natural substrate (Tocheck homogeneity of biosim

interac on with ac ve sitesame as reference biologic withreference to known substrates).

Ki with known inhibitors (1/2)(To check comparability ofcompi ve biosim interac onwith ac ve site same asreference biologic with

reference to known inhibitors).

Therapeu c Enzymes Physicochemical


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An bodies Physicochemical

Sequence and amino acidcomposi on (To check purity).

Tryp c map (1D and 2D) (Tocheck if secondary structure is

conserved).

Light and heavy chain separa on(To check an genic recogni onmo f).

IgG type (To check speci cityof IgG in localiza on of speci c

ssues/ plasma).

Purity on HPLC (RP, SEC, IEX)/MALDI (To check if prepara onis free of any impuri es).

Gel electrophoresis (IEF, SDS

PAGE and Na ve PAGE), Westernblot (To check qualita ve puritydi erence).

Absorp on spectrum from 190to 800 nm (To check purity).

An bodies Biological

Neutralizing ac vity in actual targethost cell (at least one highly prevalentIndian variant/isolate should be used) (Tocompare ac vity of similar biologic with

reference biologic in the target cell) Neutralizing ac vity in closest animal

species (if feasible) upon administra on(along with vehicle as nega ve control)(at least one highly prevalent Indianvariant/isolate should be used) (Tocompare ac vity of similar biologic withreference biologic in the target cell whenadministered in whole animal, this willevaluate the e ciency of vector/ an bodyloca on and promoter ac vity in targetcell).

Kine cs of Neutralizing ac vity duringthe proposed therapeu c period ofprotec on (at least one highly prevalentIndian variant/ isolate should be used) (Tocompare half life of the similar biologic

with reference biologic)

E cacy in appropriate disease/ infec onmodel in vitro and/or in vivo (If available)(To compare therapeu c interference andtoxicity due to a marker in the similarbiologic with that of reference biologic)

2D. Physicochemical and biological characteriza on of an bodies


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CD spectrum from 190 to 800nm (To check if secondarystructure is conserved).

Helix to Coil Transi on pro les(To verify if the prepara on isstable and impuri es or isofomsare a ec ng the stability).

Epitopic mapping of thean body binding to speci cand non-speci c epitopeswith an genic variant isolatedfrom an Indian isolates (Tocheck speci city pro le ofsimilar biologic with reference

biologic in epitope recogni on,par cularly in recogni on ofIndian variant of a host cellprotein or infec ous agentcoded protein).

An -body dilu on factors inneutraliza on (To check symbolwith reference biologic in theneutraliza on strength of thean body prepara on)

An bodies - Physicochemical


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