1. US-Nanotechnology Characterization Lab Scott McNeil, PhD Director Workshop Session 7: EU-US and International cooperation NMBP 14-2017: Regulatory Science Framework for assessment of risk benefit ratio of Nanomedicines and Biomaterials Thematic brokerage workshops Workshop 7: EU-U.S Cooperation EU Brokerage Event on Kets in Horizon 2020 Strasbourg, 1st October 2015 1 01/10/2015 - BE KETs Regulatory Science Framework for Nanomedicines and Biomaterials

2. US-NCL Mission 2 01/10/2015 - BE KETs Regulatory Science Framework for Nanomedicines and Biomaterials The NCL was established in 2004 as an interagency collaboration among NCI, NIST, and FDA. The lab’s primary mission has been to accelerate the translation of promising nanotech cancer drugs and diagnostics using a standardized “Assay Cascade. Big Pharma Academic PIs Biotech Companies Government R & D Community In Vivo Characterization In Vitro Characterization Physicochemical Characterization Clinical Regulatory Commercialization Support Assay Cascade

3. 3 01/10/2015 - BE KETs Equivalence testing for nanosimilars Addressing FDA’s scientific questions NBCDs Collaborations with Pharma, CMOs & industry consortia Immunotox Active targeting Working with instrument manufacturers Provides “pharmaceutical mentorship” for materials scientists and engineers Other indications, EHS, etc.

4. 4 01/10/2015 - BE KETs Regulatory Science Framework for Nanomedicines and Biomaterials Over 300 Nanomaterials Characterized Types of Samples NCL has characterized over 300 different nanomaterials and a wide range of various platforms. NCL has an average of 15 active collaborations at any given time. NCL characterizes an average of 75 samples each year. NCL testing is tailored to the platform properties, API, route of administration, and intended therapeutic outcome of the individual nanomedicine. NCL testing links physicochemical properties to biological outcomes.

5. 5 In clinical trials

6. 6 01/10/2015 - BE KETs Regulatory Science Framework for Nanomedicines and Biomaterials NCL-FDA Relationship NCL is trusted source for preclinical data on nanomaterials. NCL allows FDA to preview what’s in pipeline for nanotech INDs/IDEs. Quarterly interactions (visits, working groups) with FDA to maintain collaboration. Collaborations with FDA to fill gaps/better inform regulatory process: methods development, basic research and grand challenges Addressing regulatory concerns facilitates commercialization. FDA provides input on NCL’s assay cascade and is represented on NCL’s scientific oversight committee. NCL participates in FDA public meetings on topics related to nanomedicine.

7. 7 01/10/2015 - BE KETs Regulatory Science Framework for Nanomedicines and Biomaterials NCL Supports Translation/Regulation Preclinical Characterization Regulatory Concerns Clinical Characterization Exploring Alternate Indications Next-Generation Nanoparticles Historical NCL knowledge used to make recommendations in many areas, helping sponsors gain regulatory approval. Nanomedicine Developers

8. 8 01/10/2015 - BE KETs NCL-FDA Scientific Collaborations Immunological reactions to nanomaterials in non-human primates. Dermal penetration of nanomaterials in sunscreens and cosmetics. Compared endotoxin levels in experimental nanomaterial products to levels in FDA-approved products. Penetration of nanoscale TiO 2 particles in rodents, pig skin, and human skin. Characterization and purification of dendrimers for a dermal penetration study. Effects of sterilization procedures (autoclave, gamma irradiation, etc.) on silver colloids. Upcoming collaboration to study bioequivalence of follow-on nanomedicine products. Office of Generic Drugs

9. Endotoxin screening should be included whenever biological assessments of the materials are going to be performed. NCL pre-screened all ENMs used in this study. NCL Particle Designation Nanomaterial Endotoxin, Kinetic Turbidity LAL First test lots New, Large- scale lots NIEHS-1 20 nm, citrate stabilized silver 42.7 EU/mL < 0.5 EU/mL NIEHS-2 20 nm, PVP stabilized silver 113 EU/mL ≤ 2.2 EU/mL NIEHS-3 110 nm, citrate stabilized silver 0.42 EU/mL < 0.5 EU/mL NIEHS-4 110 nm, PVP stabilized silver 144 EU/mL < 0.5 EU/mL 9 Coordination between Agencies

10. Characterization in pristine vs. biological setting matters. BET = Brunauer, Emmet, and Teller 10 Vendor used BET for measuring size – BET is a surface area measurement, not accurate for size measurements. Formula: CeO 2 Purity: 99.5% minimum (based on rare earth oxide impurities) Formula Weight: 172.12 g/mol Melting Point: 2600°C Density: 7.132 g/mL Form: 15-30 nm average particle size, powder Manufacturer-Stated Specs: What the Material Actually Looks Like: Micron-sized aggregates/agglomerates Largely insoluble in aqueous media Pristine vs. Biological Settings

11. Transmission EM Scanning EM Centrifugal Field-Flow Fractionation CNTs will exist in a variety of sizes, shapes, and agglomeration states. Vendor specs: OD 10-20 nm, length 0.5-2 μm Vendor specs Reality 11 Manufacture vs. Experimental

12. 12 Need better protocols for dispersing poorly soluble ENMs; better reproducibility between labs. Many labs will sonicate and use immediately, (hopefully) before ENMs settle out. Some add coatings to help solubilize, but can these influence other factors. Problem: Poorly soluble ENMs This is an important step in correlating PCC to biological properties. Recommendations

13. 13 Very limited PCC and size tools are available for many poorly soluble ENMs. Most require an aqueous resuspension. For example, MWCNTs typically only report TEM measurements on a very select population. Problem: Limited characterization tools Need consistency across labs in measuring and reporting PCC. How should size of polydispered samples be reported? Size by XRD vs TEM vs DLS can all be very different. Problem: Size reporting for polydispersed samples Recommendations

14. 29/11/2013 - BE KETsAdressed challenge/PPP14 Systematic investigations of the mechanisms and effects of engineered nanomaterial interactions with living systems and/or the environment  36 partners across the globe working together to study more than two dozen different ENMs NCL’s Role, Testing 15 ENMs Physicochemical Characterization Size/Size Distribution DLS TEM Surface Charge Zeta Potential Composition EDS In Vitro Cytotoxicity MTT Cell Viability Assay LDH Membrane Integrity Assay Porcine Renal Proximal Tubule (LLC-PK1) Cells Human Hepatocellular Carcinoma (HepG2) Cells

15. 15 01/10/2015 - BE KETs Regulatory Science Framework for Nanomedicines and Biomaterials Cooperation with EU-NCL Expansion of resources for nanomedicine developers. Expanded visibility Greater quality control & harmonization Reduced risk of an adverse event in nanomedicine EU-NCL will facilitate interaction with EMA NCL will work closely with EU-NCL to establish SOPs, keep lines of communication open.

16. Contact Person: Scott McNeil, PhD Director OrganisationNanotechnology Characterization Laboratory Address P.O. Box B Frederick, MD 21703 United States Phone nr(301) 846-6939 E-mailncl@mail.nih.gov 16 01/10/2015 - BE KETs Regulatory Science Framework for Nanomedicines and Biomaterials Contact Details