Evolution of Specifications and Analytical Methods During ...

1 Evolution of Specifications and Analytical Methods During Synthetic Peptide Drug DevelopmentMichael Verlander, , Proactive quality ComplianceBiologics Monographs 1 Peptides & Insulins Expert Committee MemberWebinar, March 14, 20192 2017 USP Definitions Challenges Regulatory guidance Specifications quality attributes/ Specifications for peptides Characterization Methods Qualification/validation requirements Peptide-related impurities Origins Regulatory guidance Examples of Purity/Related Substances method development Setting limits for peptide-related impurities Bioassay/bioidentityconsiderations Final thoughtsAgenda3 2017 USP Protein Any alpha amino acid polymer with a specific defined sequence that is greater than 40 amino acids in size (May be)

2 Regulated as biological products or drugs, , insulin and human growth hormone) Chemically synthesized polypeptide Any alpha amino acid polymer that is (a) made entirely by chemical synthesis, and (b) is less than 100 amino acids in size (Regulated as drugs unless they meet the statutory definition of biological products ) Peptide Any alpha amino acid polymer with specific defined sequence composed of 40 or fewer amino acids (Regulated as a drugs unless they meets the statutory definition of biological products , peptide vaccines)FDA Draft Guidance.

3 Biosimilars: Questions and Answers Regarding Implementation of the Biologics Price Competition and Innovation Act of 2009. February 2012 Definitions4 2017 USP Biological product A virus, therapeutic serum, toxin, antitoxin, vaccine, blood, blood component or derivative, allergenic product, protein (except any chemically synthesized polypeptide), or analogous product, or arsphenamine or derivative of arsphenamine (or any other trivalent organic arsenic compound), applicable to the prevention, treatment, or cure of a disease or condition of human beings Proposed Rule: Definition of the Term Biological Product.

4 Federal Register, Vol. 83, No. 238. Wednesday, December 12, 2018 Definitions (Continued)5 2017 USP Because of improved manufacturing technology, the complexity of synthetic peptides entering clinical development is increasing: Longer sequences (up to 50 AAs or more) Macrocyclic/multicyclic structures Conjugates with lipids, PEG, proteins, etc. Characterization of the drug substances and their related impurities becomes correspondingly more challenging In order to keep pace with the clinical development program, the manufacturing process and the associated Analytical Methods (in-process and final product) must be developed in parallel Because of their complexity, a relatively large number of tests is required to adequately describe the quality characteristics of peptides (the specification )

5 Challenges6 2017 USP Additional testing is performed During characterization studies (ongoing throughout development) Meeting regulatory requirements presents some challenges, since synthetic peptides are specifically excluded from most guidance documents, particularly those for product-related impuritiesChallenges (Continued)7 2017 USPICH Q6A Specifications : Test Procedures and Acceptance Criteria for New Drug Substances and New Drug Products: Chemical Substances specification : A specification is defined as a list of tests, references to Analytical procedures, and appropriate acceptance criteria which are numerical limits, ranges, or other criteria for the tests described.

6 It establishes the set of criteria to which a new drug substance or new drug product should conform to be considered acceptable for its intended use. The following tests and acceptance criteria are considered generally applicable to all new drug substances: Description: A qualitative statement about the state ( solid, liquid) and color of the new drug substance. Regulatory Guidance Specifications8 2017 USPI dentification: Identification testing should optimally be able to discriminate between compounds of closely related structure which are likely to be present.

7 Identification tests should be specific for the new drug substance, , infrared spectroscopy. Assay: A specific, stability-indicating procedure should be included to determine the content of the new drug substance. In many cases it is possible to employ the same procedure ( , HPLC) for both assay of the new drug substance and quantitation of impurities. Impurities: Organic and inorganic impurities and residual solvents are included in this category. Regulatory Guidance Specifications (Continued)9 2017 USPG uidance for Industry. CGMP for Phase I Investigational Drugs (US FDA, 2008) You should perform laboratory testing of the phase 1 investigational drug to evaluate quality attributes including those that define the identity, strength, potency, purity, as appropriate.

8 Specified attributes should be monitored, and acceptance criteria applied appropriately. For known safety-related concerns, Specifications should be established and met. Guidance for Industry. INDs for Phase 2 and Phase 3 Studies. Chemistry, Manufacturing and Controls Information (US FDA, 2003) During the clinical investigation process, the sponsor would establish tentative acceptance criteria that are continually refined based on data obtained from analysis of batches of drug substance and new information that becomes available. In the course of product development, the Analytical technology or methodology often evolves parallel to the clinical investigations.

9 In setting subsequent acceptance criteria, relevant correlations should be established between data generated During early and late drug development. Regulatory Guidance Specifications (Continued)10 2017 USPQ uality AttributeMethodCommentsAppearanceVisual inspectionIdentificationMass spectral analysisAmino acid analysisHPLC comparison with standard (co-elution)Peptide mappingMonoisotopic massHydrolysis and derivatization Methods should be specifiedMethod may be same as related substancesOnly rarely required (for complex sequences)AssayHPLC Preferred method if quantitative standard availablePeptide ContentQuantitative amino acid analysisNitrogen determinationHydrolysis method must be validated; only stable amino acids used for calculationElemental analysis preferred.

10 Kjeldahland HPLC using chemiluminescence nitrogen detector (CLND) may also be usedRoutine quality Attributes for Peptide APIs11 2017 USPQ uality AttributeMethodCommentsOrganic ImpuritiesPeptide-related Impurities, including aggregates (for more complex sequences)HPLC Methods (RP-HPLC, IEX-HPLC, SEC-HPLC, etc.)UHPLC Methods preferred; must be validated for both process-related and degradation impurities; limits for individual and total impurities should be specifiedNon-peptide Impurities:Residual solventsResidual TFAGas chromatographyRP-HPLC, Ion chromatographyMay be limited to those used in the final process steps (after validation)Only if used in manufacturing processInorganic ImpuritiesOther anionsIon chromatographyOnly those used in manufacturing process ( , phosphate)FluorideIon selective electrode orion chromatographyOnly if used in manufacturing processRoutine quality Attributes for Peptide APIs (Continued)12 2017 USPQ uality AttributeMethodCommentsInorganic Impurities (Continued)