Dr. Javier Polo Vice President Research & Development

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Efecto de l suplementción con concentrdo de inmunoglobulins prtir del suero bovino en l Microbiot y su contribución l slud intestinl y l sistem nervioso centrl Dr. Jvier Polo Vice President Reserch & Development

Indice GRUPO LGI CONCENTRADO DE SUERO BOVINO INMUNOGLOBULINAS ESTUDIOS CLINICOS EN PACIENTES DE IBS/IBD MECANISMO DE ACCION EFECTOS EN LA MICROBIOTA EFECTOS EN LA SALUD GASTRO INTESTINAL EFECTOS EN LA SALUD DEL SISTEMA NERVIOSO CENTRAL

Vegetl & Animl Helth/Nutrition Humn Nutrition Phrm/Biologicls/Humn helth Rw mteril Blood, Bones, Fether, Ftty tissues ll by-products Blood Diry Blood, Skin, Ftty Tissue, Bones Blood Blood The LGI Group serves industril, veterinry nd humn pplictions with high qulity, nturl source proteins.

Medicl Food Product infectious Cncer Cchexi Other DI IBS IBD Specilties Trgeted: HIV Enteropthy Norovirus nd Cryptosporidium C. Difficle Cncer Associted weight Loss Syndrome Gut Brrier Dysfunction nd microbil trnsloction Chemotherpy Induced Mucocitis Post-Infectious Dirrhe Peditric IBS-D Infnt Dirrhe Symptom Mngement Smll Intestine Bcteril Overgrowth GI Anti-inflmmtory Hypercholesterolemi Chron's Disese Ulcertive Colitis Peditric UC Pouchitis Celic nd Gluten Sensitivity Antibiotic Induced Dirrhe DMARD-Induced Dirrhe Chronic Mesenteric Ischemi Lymphocytic Colitis Chronic Dirrhel Ileorectl Anstomosis Fecl Incontinence Mstocytic Enterocolitis Short Bowel Syndrome Microbiome Dysbiosis Autism Heptic Leky GUT Acid Reflux H. Pylori

Clinicl Tril Results IBS-D Pilot study results suggest tht EnterGm provides for distinctive nutritionl requirement unique for IBS-D ptients which could not be received from soy protein. Clinicl Medicine Insights: Gstroenterology Evlution of Serum-Derived Bovine Immunoglobulin Protein Isolte in Subjects with Dirrhe-Predominnt Irritble Bowel Syndrome - 2013:6 49 60. Dle Wilson, Mlknthi Evns, Eric Wever, Audrey L. Shw, Gerld L. Klein https://www.ncbi.nlm.nih.gov/pmc/rticles/pmc4020402/ * Results from 6-week, rndomized, double-blind, plcebo-controlled study (N = 66). Subjects received either EnterGm 10 g dily, 5 g dily, or plcebo. Subjects self-ssessed presence nd severity of GI symptoms dily using 4-point scle (0 = none, 1 = mild, 2 = moderte, 3 = severe). Number of dys with symptoms ws clculted for ech 2-week period per subject. Dt represents within-group chnges from bseline to end of study mong: (1) Subjects tking 10 g dose of EnterGm dily (n=15); nd (2) subjects within sy protein plcebo group (n=22). No sttisticlly significnt reduction in the number of dys with ny GI-relted symptoms for plcebo group. Study ws not powered to demonstrte between-group differences. PRIVATE AND CONFIDENTIAL

Published Clinicl Cses Irritble Bowel Syndrome w/ Dirrhe (IBS-D) PRIVATE AND CONFIDENTIAL

PRIVATE AND CONFIDENTIAL Published Clinicl Cses Inflmmtory Bowel Disese (IBD)

Mode of Action https://www.youtube.com/wtch?v=zf4c-plk908

Effect of SDP/SBI supplementtion in Aging Model INTESTINAL EFFECTS 2 Month-old 6 Month-old Dietry supplementtion with SDP will reduce the mgnitude of intestinl dysfunction ssocited with ging On diet (4 months) SYSTEMIC EFFECTS SAMP8 2M 6M-CTL 6M-SDP Effects of SDP supplementtion on inflmmtory mrkers & plsm mino cid profile CENTRAL NERVOUS SYSTEM EFFECTS SDP supplementtion my reduce the inflmmtory stte of the CNS during ging nd prevents the cognitive dysfunctions 9

Fold chnge Intestinl Effect on Senescent Mice Fold chnge Fold chnge Intestinl effects Pro-inflmmtory cytokines Men ± SEM (N = 5-8 mice/group) b, P<0.05 3 2 1 TNF-α expression b 3 2 1 IFN-γ expression b 3 2 1 IL-6 expression b + SDP 0 Conclusions 2M 6M 6M-SDP 0 2M 6M 6M-SDP 0 2M 6M 6M-SDP Six month-old SAMP8 mice showed low-grde inflmmtory stte in the jejunum mucos. SDP supplementtion ws ble to prevent this effect, confirming observtions from other inflmmtory models (SEB-Intestine, Colitis or LPS-Lung models)

pg IL-6/mL Systemic Effects on Senescent Mice pg TNF-α/mL pg IL-10/mL Systemic effects Pro-inflmmtory cytokines Men ± SEM (N = 5-6 mice/group) b, P<0.05 Anti-inflmmtory cytokines 0,4 0,2 IL-6 in Serum b 10 5 TNF-α in Serum b 0,15 0,10 0,05 IL-10 in Serum b + SDP 0,0 2M 6M-CTL 6M-SDP 0 2M 6M-CTL 6M-SDP 0,00 2M 6M-CTL 6M-SDP Conclusions Six month-old SAMP8 mice showed systemic low-grde inflmmtory stte. As previously oberved in other inflmmtory models (SEB-Intestine, Colitis or LPS-Lung models), SDP supplementtion ws ble to prevent this effect

Effects on the Centrl Nervous System SDP /SBI 12

Collins MS et l (2012) Nture Rev Microbiol 10, 735-742 Effects on the Centrl Nervous System Hormones GALT cells Cytokines Bcteril products Neurons MICROBIOTA SDP/SBI Externl environment 13

Biologicl Functionlity of Serum frctions, Summry Intestinl effects Systemic effects CNS effects Immunosenesence Unspecific response Pro-inflmmtory cytokines Anti-inflmmtory cytokines Cognitive decline improvement Blood Brin Brin Brrier Brrier Pro-inflmmtory cytokines + SDP/SBI GUT-BRAIN AXIS Adhesion molecules Rective Oxidtive Species

Concluding Comments APC nd Enter Helth s Serum proteins re widely used in humns nd other mny species, including compnion nimls It is recognized tht these functionl proteins re mixture of proteins tht mitigte the negtive effects of inflmmtion Used ppropritely, functionl proteins cn be tool to improve humns nd niml well being