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Copyright © 2019 DuPont or its affiliates. All Rights Reserved. The DuPont Logo Oval and DuPont™ are trademarks or registered trademarks of E. I. du Pont de Nemours and Company. Samples manufactured in DuPont application laboratories may contain allergens. The information contained herein is based on data known to DuPont or its affiliates at the time of preparation of the information and believed by them to be reliable. This is business-to-business information intended for food, beverage and supplement producers, and is not intended for the final consumer of a finished food, beverage or supplement product. The information is provided “as is” and its use is at the recipient’s sole discretion and risk. It is the recipient’s sole responsibility to determine the suitability and legality of its proposed use of DuPont products for its specific purposes. Information and statements herein shall not be construed as licenses to practice, or recommendations to infringe, any patents or other intellectual property rights of DuPont or others. DUPONT HEREBY EXPRESSLY DISCLAIMS (I) ANY AND ALL LIABILITY IN CONNECTION WITH SUCH INFORMATION, INCLUDING, BUT NOT LIMITED TO, ANY LIABILITY RELATING TO THE ACCURACY, COMPLETENESS, OR USEFULNESS OF SUCH INFORMATION, AND (II) ANY AND ALL REPRESENTATIONS OR WARRANTIES, EXPRESS OR IMPLIED, WITH RESPECT TO SUCH INFORMATION, OR ANY PART THEREOF, INCLUDING ALL REPRESENTATIONS AND WARRANTIES OF TITLE, NONINFRINGEMENT OF COPYRIGHT OR PATENT RIGHTS OF OTHERS, MERCHANTABILITY, FITNESS OR SUITABILITY FOR ANY PURPOSE, AND WARRANTIES ARISING BY LAW, STATUTE, USAGE OF TRADE OR COURSE OF DEALING. INTRODUCTION: Obesity is a top global health concern, and the gut microbiota and energy balance are interlinked. HOWARU® Shape, Bifidobacterium animalis ssp. lactis 420 (B420™) consumed with or without fiber polydextrose (Litesse® Ultra™, LU), offers a promising candidate to be utilized in the combat against obesity epidemic. METHODS: The efficacy of HOWARU ® Shape on metabolic health has been studied in preclinical cell models, mouse models and in a clinical trial (Fig 1). PROBIOTIC Bifidobacterium animalis ssp. lactis 420™ FOR METABOLIC HEALTH Uusitupa Henna-Maria 1* , Lehtoranta Liisa 1* , Maukonen Johanna 1* 1 DuPont Nutrition and Biosciences, Kantvik, Finland DuPont Nutrition & Biosciences Danisco Sweeteners Oy Sokeriehtaantie 20 02460 KANTVIK FINLAND Email: [email protected] www.dupontnutritionandhealth.com RESULTS: In cell model, B420™ supernatant alone (Fig 2) 1,2 , but not in combination with fish oil 2 , was shown to improve epithelial integrity. As intestinal barrier function has been associated with an increasing variety of diseases – both intestinal and systemic – this finding led to hypothesis that HOWARU® Shape, could benefit metabolic health via increasing tight junction integrity. In following diabetogenic and obesogenic mouse models, B420™ with or without LU was shown to reduce weight gain during HFD while sustaining the intestinal integrity (Fig 3), reducing glucose levels and improving insulin sensitivity 3,4,5,6 . In a clinical trial, B420™ controlled body fat mass (compared to placebo), mostly in the abdominal region (Fig 4) 7 . Body fat mass results were associated with serum zonulin levels 7 , supporting the preclinical findings of B420™ enhancing the epithelial integrity 1,2,4 . Further, B420™ alone and with LU modified gut microbiota by increasing the abundance of bacteria associated with a lean phenotype such as Akkermansia spp., Christensenellaceae spp compared to baseline 8 . The increase in Christensenellaceae spp was associated to clinical outcomes (Fig 5) 8 . CONCLUSIONS: Both preclinical and clinical data suggests that HOWARU® Shape is a strong candidate in improving metabolic health, especially in weight management. Through its capability to reduce intestinal permeability and to modulate gut microbiota composition leading possibly to subsequential slow down of detrimental processes associated to endotoxemia and low-grade inflammation in obese states HOWARU® Shape offers an exiting probiotic for weight management. References: 1. Putaala et al. Research in Microbiology 2008; 159:69 2. Mokkala et al. Nutr Res. 2016;36(3):246; 3. Amar et al. EMBO Mol Med 2011;3(9):559; 4. Stenman et al. Benef Microbes 2014;5:437; 5. Stenman et al. Diabetol Metab Syndr 2015;7:75; 6. Garidou et al. Cell Metab. 2015;22(1):100-12; 7. Stenman et al. EBioMedicine. 2016 Nov;13:190-200; 8. Hibberd et al. Benef Microbes. 2019;10(2):121-135. Fig. 1. Preclinical in vitro and animal trials as well as the clinical trials completed or ongoing with HOWARU® Shape and metabolic health outcomes. Fig 2. B420™ supernatant improves epithelial integrity in a Caco-2 cell model. FIg3. In diabetogenic mouse model, B420™ was able to ameliorate the disruption of gut barrier integrity measured by LPS in circulation. Fig 4. In per protocol population of clinical trial HOWARU® Shape resulted in 4.5% (1.4 kg) less fat mass vs. placebo (P=0.02). Fig 5. The microbiota results of the clinical trial indicate that HOWARU® Shape modifies positively gut microbiota composition in 6 months intervention, and the modifications correlate with clinical outcomes. Relative abundance (mean ± SD) of Christensenellaceae for Placebo, LU, B420 and LU+B420 groups (a). Spearman correlations between Christensenellaceae abundance and Waist hip ratio (b) and Energy intake (c) at baseline and for dual-energy X-ray absorptiometry (DXA) Android fat (d) and Trunk fat (e) after 6 mo of intervention with LU+B420. P<0.01, **P<0.01, *P<0.05 vs Placebo, Kruskal-Wallis with Steel-Dwass post-hoc test and Benjamini- Hochberg FDR correction. 0 1 2 3 4 5 6 7 NC Control B420 a b a Plasma LPS (EU/ml) High fat, low carb diet • Caco-2 cells are treated with cell-free supernatants of four probiotics including B420™ after cultivating the bacteria anaerobically at 37 Celsius in Man, Rogosa and Sharpe (MRS) broth supplemented with 1.0% glucose and differentiating the Caco-2 cells by short-term differentiation protocol. Then, Caco- 2-cells were incubated for 24 hours with cell-free supernatants of the tested strains, and TEER measurement was performed 1 • In another study the impact of cell-free culture supernatant of B420™ with and without fish oil on intestinal permeability in a CaCo-2 cell model was studied 2 Diabetogenic mouse model 3 : Mice were fed a high-fat, ketogenic diet (72 energy % fat) for 4 weeks, with a 6-week subsequent treatment with B420™ (10 8 -10 10 cfu/day) or vehicle Obesogenic mouse model: C57Bl/6J mice were fed a high-fat diet (60 energy %) for 12 weeks, and gavaged daily with B420™ (10 9 cfu) or vehicle. • A randomized placebo-controlled multicenter clinical trial with a 6 months intervention • Study design and execution follows ICH-GCP • 225 Finnish participants with BMI 28-34.9, otherwise healthy • Primary outcome: relative change in body fat mass, Secondary and exploratory parameters: body composition and metabolic health outcomes, gut microbiota composition and metabolites • Four arms: 1. Placebo, 2. B420™ (10 10 cfu/day), 3. PDX (12 g), 4. B420™ (10 10 cfu/day) with LU • Clinicaltrials.gov: NCT01978691 *Confilct of interest: All authors work at DuPont Nutrition and Biosciences. New clinical trial initiated 2019 Mokkala et al. Nutr Res 2016 2 Putaala et al Res Microbiol 2008 1 Amar et al. EMBO Mol Med 2011 3 Stenman et al. EBiomedicine 2016 7 Stenman et al. Diabetol Metab Synd 2015 5 Garidou et al. Cell Metab 2015 6 In preclinical in vitro models: In obesogenic and diabetogenic mouse models: In a clinical trial: Fig 2 Fig 3 Fig 1 Fig 4 Hibberd et al. Benef Micr 2019 8 Stenman et al. Benef Micr 2014 4 Fig 5
