CHARACTERIZATION AND PRE-CLINICAL STUDY OF A FRENCH WEST COAST (FWC) PROPOLIS EXTRACT

Transcription

1 CHARACTERIZATION AND PRE-CLINICAL STUDY OF A FRENCH WEST COAST (FWC) PROPOLIS EXTRACT B. Poirot a, V. Léonard-Nevers a, H. Hachet b, G.Grellier b. a APINOV, Pôle technologique 40 rue Chef de Baie La Rochelle, FRANCE Tel: +33 (0) , Fax: +33 (0) contact@apinov.com b NAT INOV, 13 rue Belle Vue Saint Lezin, FRANCE Tel: +33 (0) , Fax: +33 (0) , DIRECCTE POITOU-CHARENTE 1

2 COMPANY APINOV is an engineering office specialized in international beekeeping issues Our mission: coordinate the execution of innovating projects, technical studies and Research and Development plans 2

3 COMPANY Our interlocutors: beekeepers, industrialists and laboratories Our team: -2 PhD s in Biochemistry/Biotechnology -1 Food-processing engineer, specialized in sensory tests -1 Responsible for market studies, communication & international affairs Our goal: promote beekeeping by developing new techniques and innovating products 3

4 CHARACTERIZATION AND PRE-CLINICAL STUDY OF A FRENCH WEST COAST (FWC) PROPOLIS EXTRACT 4

5 PROBLEMATIC PROPOLIS has a lot of biological activities (antibiotic, antiviral, antifungal, anti-germinating, extremely powerful anesthetic, healing by stimulation of tissue regeneration, etc) Variability in chemical composition of propolis, which depends of its geographical and botanical origins, is limiting its application for human and veterinary medicine A solution could be to create standardized propolis extract in order to ensure a chemical composition and consequently biological properties reproducibility 5

6 SCIENTIFIC OBJECTIVES 1. To create a standardized French propolis extract 2. To characterize the standardized propolis extract 3. To validate the propolis extract safety by a preclinical study 6

7 EXPERIMENTAL French propolis = European propolis = Poplar propolis Different landscapes in France Landscape French West Coast (FWC) 7

8 EXPERIMENTAL French West Coast (FWC): 3 major ecosystems: marshland grove forest / Propolis mix: For each ecosystem, a sample of propolis was prepared with a mix of at least 10 different apiaries (32 hives per apiary) 8

9 EXPERIMENTAL Extraction 1. Raw propolis was grinded to obtain a powder 2. Polyphenolic fraction was extracted by maceration in a hydroalcoholic solution at 70% ethanol during 24 hours and under stirring (traditional method) 3. Propolis extract was dried 9

10 EXPERIMENTAL Characterization The polyphenolic composition of the extract was determined : qualitatively by HPLC-MS quantitatively by HPLC-DAD for : caffeic acid, Coumaric acid, Pinocembrin and Galangin By a total quantification using the Folin-Ciocalteu method 10

11 Pre-clinical study EXPERIMENTAL - Cytotoxicity tests on skin explants - Ocular tolerance tests on fibroblasts from rabbit cornea - Phototoxicity tests on balb/c 3T3 mouse fibroblasts 11

12 RESULTS: qualitative composition of raw propolis Pinocembrin Galangin Marshland Grove Forest 12

13 RESULTS: quantitative composition of raw propolis Subtype of propolis Pinocembrin concentration (mg.l -1 ) Galangin concentration (mg.l -1 ) Grove propolis 17 7 Marshland propolis 45 2 Forest propolis

14 RESULTS: standardization FWC Propolis extract = mix of the 3 subtypes 14

15 RESULTS: characterization 15

16 RESULTS: characterization Composition of FWC propolis extract is original and unique Compounds (% in the extract) Biological activities Galangin 8.8 antioxidant and vasorelaxant activities Pinocembrin 7.0 antimicrobial, antifungal, antiinflammatory and neuroprotective effect P-coumaric acid 1.6 Anti ulcer Caffeic acid 0.5 Anti ulcer Ferulic acid Nd _ Isoferulic acid Nd _ Pinobanksin-5-methyl ether Nd _ Pinobanksin Nd _ Kaempferol Nd _ Caffeic acid phenethyl ester (CAPE) Nd _ Pinobanksin-3-O-butyrate Nd _ Pinobanksin-3-O-pentanoate Nd _ 16

17 RESULTS: characterization Total polyphenol content of the FWC propolis extract : 17% => High quality propolis (Gardana, 2007) 17

18 RESULTS: pre-clinical study Objective: Validation of propolis extract safety Cytotoxicity Ocular tolerance Phototoxicity 18

19 RESULTS: pre-clinical study Cytotoxicity: no cytotoxicity was observed for propolis extract topically applied on skin explants (for all tested concentrations). 19

20 RESULTS: pre-clinical study Ocular tolerance: very low irritating effect on fibroblasts. Phototoxicity: very low phototoxic effect. 20

21 CONCLUSION French West Coast (FWC) propolis extract: standardized propolis extract High quality product Safety product Clinical study in progress 21

22 ACKNOWLEDGMENT Apinov Team Beekeepers Financial Partners Companies DIRECCTE POITOU-CHARENTE 22

23 THANK YOU FOR YOUR ATTENTION 23