GRAS Notice (GRN) No. 922 https://www.fda.gov/food/generally-recognized-safe-gras/gras-notice-inventory March 19, 2020 Rachel Morissette, Ph.D. Regulatory Review Scientist Center for Food Safety and Applied Nutrition Office of Food Additive Safety U.S. Food and Drug Administration CPK-2 Building, Room 2092 5001 Campus Drive, HFS-225 College Park, MD 20740 Dear Dr. Morissette: It is our opinion that the enclosed GRAS Determination for the Use of 6’-Sialyllactose Sodium Salt (6’-SL) in Non-Exempt Term Infant Formula constitutes a new notification. Although 6’- SL is the subject of other GRAS Notices, the subject of the enclosed GRAS Notice is produced using a novel production process. We thank you for taking the time to review this GRAS determination. Should you have additional questions, please let us know. Sincerely, Dietrich B. Conze, Ph.D. Managing Partner Enclosure: CD containing Form 3667, cover letter, GRAS Determination for the Use of 6’-Sialyllactose Sodium Salt (6’-SL) in Non-Exempt Term Infant Form, and all references Spherix Consulting Group, Inc. 11821 Parklawn Drive, Suite 310, Rockville, MD 20852 301-557-0375; [email protected] GRAS Determination for the Use of 6’-Sialyllactose Sodium Salt in Non-Exempt Term Infant Formula Prepared for: Jennewein Biotechnologie GmbH Maarweg 32 D-53619 Rheinbreitbach Germany Prepared by: Spherix Consulting Group, Inc. 11821 Parklawn Drive, Suite 310 Rockville, MD 20852 March 19, 2020 GRAS Notification for the Use of 6’-Sialyllactose Sodium Salt March 19, 2020 Prepared for Jennewein Biotechnologie GmbH TABLE OF CONTENTS I. SIGNED STATEMENT OF THE CONCLUSION OF GENERALLY RECOGNIZED AS SAFE (GRAS) AND CERTIFICATION OF CONFORMITY TO 21 CFR §170.205-170.260 .... 1 A. SUBMISSION OF GRAS NOTICE .................................................................................1 B. NAME AND ADDRESS OF THE SPONSOR ................................................................1 C. COMMON OR USUAL NAME .......................................................................................1 D. TRADE SECRET OR CONFIDENTIAL INFORMATION ............................................1 E. INTENDED USE ..............................................................................................................1 F. BASIS FOR GRAS DETERMINATION .........................................................................1 G. PREMARKET APPROVAL ............................................................................................4 H. AVAILABILITY OF INFORMATION ...........................................................................4 I. FREEDOM OF INFORMATION ACT (FOIA) ...............................................................4 J. INFORMATION INCLUDED IN THE GRAS NOTIFICATION...................................4 II. IDENTITY, METHOD OF MANUFACTURE, SPECIFICATIONS, AND PHYSICAL OR TECHNICAL EFFECT OF THE NOTIFIED SUBSTANCE ........................................................ 1 A. COMMON OR USUAL NAME .......................................................................................1 B. CHEMICAL NAME .........................................................................................................1 C. MOLECULAR FORMULA AND MASS ........................................................................1 D. STRUCTURAL FORMULA ............................................................................................1 E. DESCRIPTION OF 6’-SL ................................................................................................1 F. PRODUCTION PROCESS ...............................................................................................2 1. Description of the Production Strains ........................................................................... 2 2. Manufacturing Process ................................................................................................. 5 G. FINISHED PRODUCT SPECIFICATIONS AND OTHER QUALITY ATTRIBUTES 6 1. 6’-SL Product Specifications and Batch Data Compliance .......................................... 6 2. Other Quality Attributes ............................................................................................... 8 H. STABILITY OF 6’-SIALYLLACTOSE ..........................................................................8 1. Genetic Stability of the Production Strains .................................................................. 8 2. Stability of the 6’-SL .................................................................................................... 9 III. DIETARY EXPOSURE ........................................................................................................ 11 A. INTENDED EFFECT .....................................................................................................11 B. HISTORY OF EXPOSURE ............................................................................................11 C. INTENDED USE ............................................................................................................15 D. ESTIMATED DAILY INTAKE .....................................................................................15 -ii- SPHERIX CONSULTING GROUP, INC. GRAS Notification for the Use of 6’-Sialyllactose Sodium Salt March 19, 2020 Prepared for Jennewein Biotechnologie GmbH IV. SELF-LIMITING LEVELS OF USE .................................................................................... 16 V. COMMON USE IN FOOD BEFORE 1958 ........................................................................... 17 VI. NARRATIVE ON THE CONCLUSION OF GRAS STATUS ............................................ 18 A. SAFETY OF THE PRODUCTION ORGANISM ..........................................................20 B. ABSORPTION, DISTRIBUTION, METABOLISM, AND EXCRETION ...................21 C. TOXICOLOGY ...............................................................................................................22 1. Genotoxicity ............................................................................................................... 23 2. Toxicity Studies on 6’-SL as a Single Ingredient ....................................................... 29 3. Toxicity Studies on Jennewein’s 6’-SL as Part of an HMO Mixture ......................... 32 D. TOLERANCE STUDY IN NEONATAL PIGLETS ......................................................38 1. Introduction ................................................................................................................ 38 2. Materials and Methods ............................................................................................... 38 3. Results ........................................................................................................................ 42 4. Discussion ................................................................................................................... 70 E. CORROBORATIVE ANIMAL STUDIES ....................................................................70 F. CLINICAL STUDIES .....................................................................................................71 G. ALLERGENICITY .........................................................................................................72 H. REGULATORY APPROVALS ACROSS THE WORLD ............................................73 VII. SUPPORTING DATA AND INFORMATION................................................................... 74 A. REFERENCES ................................................................................................................74 B. EXPERT PANEL STATEMENT ...................................................................................87 LIST OF TABLES Table 1. Genetic Manipulations in the Basic Strain ...................................................................... 3 Table 2. Genetic Manipulations in JBT-6SL .................................................................................. 4 Table 3. Product Specifications and Batch Data for 6’-Sialyllactose ............................................ 7 Table 4. Elemental Analysis of 6’-Sialyllactose ............................................................................ 8 Table 5. Stability of 6’-Sialyllactose as a Component of a Mixed Human Milk Oligosaccharide Powder Under Ambient Conditions (25°C, 60% Relative Humidity) .......................................... 10 Table 6. Stability of 6’-Sialyllactose as a Component of a Mixed Human Milk Oligosaccharide Powder Under Accelerated Conditions (40°C, 75% Relative Humidity) ..................................... 10 Table 7. Studies Determining the Concentration of 6’-Sialyllactose in Human Breast Milk ...... 12 Table 8. Comparison of Jennewein’s 6’-Sialyllactose Sodium Salt With the 6’-Sialyllactose Sodium Salt Tested Phipps et al., 2019 and That Supports GRN 881 .......................................... 23 -iii- SPHERIX CONSULTING GROUP, INC. GRAS Notification for the Use of 6’-Sialyllactose Sodium Salt March 19, 2020 Prepared for Jennewein Biotechnologie GmbH Table 9. Bacterial Reverse Mutation Test Performed with an HMO Mixture Containing 4.0% 6- Sialyllactosec ................................................................................................................................. 27 Table 10. In vitro Micronucleus Test in Human Peripheral Blood Lymphocytes Exposed to an HMO Mixture Containing 4.0% 6’-Sialyllactoseb ........................................................................ 29 Table 11. Statistically Significant Differences in Clinical Chemistry Values on Day