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(12) United States Patent (10) Patent No.: US 8,257,695 B2 Rautonen Et Al

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(12) United States Patent (10) Patent No.: US 8,257,695 B2 Rautonen Et Al US008257695B2 (12) United States Patent (10) Patent No.: US 8,257,695 B2 Rautonen et al. (45) Date of Patent: Sep. 4, 2012 (54) METHOD FORMODULATING SATIETY Backhed, Fredrik, et al., “Host-bacterial mutualism in the human SIGNALING WITH SPECIFIC STRANS OF intestine.” Science (2005) vol. 307 pp. 1915-1920. Backhed, Fredrik, et al., “The gut microbiota as an environmental LACTOBACILLUS ACIDOPHILUS AND factor that regulates fat storage.” PNAS (2004) vol. 101:44 pp. 15710 BACILLUS 15723 Bleau, C., et al., “New Lactobacillus acidophilus isolates reduce the (75) Inventors: Nina Rautonen, Espoo (FI); Heli release of leptin by murine adipocytes leading to lower interferon gamma production.” Clinical and Experimental Immunology (2005) Putaala, Upinniemi (FI); Arthur vol. 140:3 pp. 427-435. Ouwehand, Inkoo (FI); Kirsti Tiihonen, Fosset, Spohie, et al., “Pharmacokinetics and feeding responses to Helsinki (FI); Marta Korczynska, muramyldipeptide in rats.” Physiolgoy & Behavior (2003) vol. 79 pp. Wageningen (NL); Wouter Herman 173-182. Gee, Jennifer M., et al., “Dietary lactitol fermentation increases Noordman, Ede (NL) circulating peptide YY and glucagon-like peptide-1 in rats and humans.” Nutrition (2005) vol. 21:10 pp. 1036-1043. (73) Assignee: DuPont Nutrition Biosciences ApS, Korbonits, M., et al., “Ghrelin and cannabinoid interactions on food Copenhagen (DK) intake.” Endocrine (2005) (Abstract Only). Le Roux, C.W., et al., "Attenuated peptide YY release in obese (*) Notice: Subject to any disclaimer, the term of this subjects is associated with reduced satiety.” Endocrinology (2005) patent is extended or adjusted under 35 pp. 1-22. Lee, Hui-Young, et al., “Human originated bacteria, Lactobacillus U.S.C. 154(b) by 558 days. rhamnosus PL60, produce conjugated linoleic acid and show anti obesity effects in diet-induced obese mice.” Biochimica and (21) Appl. No.: 12/162,160 Biophysica Acta (2006) vol. 1761:7 pp. 736-744. Ley, Ruth E., et al., “Obesity alters gut microbial ecology.” PNAS (22) PCT Filed: Jan. 28, 2007 (2005) vol. 102:31, pp. 11070-1 1075. Livak, Kenneth J., et al., “Analysis of relative gene expression data (86). PCT No.: PCT/B2007/OO1186 using real-time quantitative PCR and the 2^T method.” Methods (2001) vol. 25 pp. 402-408. S371 (c)(1), Naruszewcz, Marek, et al., “Effect of Lactobacillus plantarum 299v (2), (4) Date: Sep. 18, 2008 on cardiovascular disease risk factors in Smokers.” American Journal of Clinical Nutrition (2002) vol. 76:6. pp. 1249-1255. Renshaw, D., et al., “Peptide YY: A potential therapy for obesity.” (87) PCT Pub. No.: WO2007/085970 Current Drug Targets (2005) vol. 6:2, pp. 171-179 (Abstract Only). PCT Pub. Date: Aug. 2, 2007 Roberfroid, Marcel B., “Prebiotics and probiotics: are they functional foods?.” American Journal of Clinical Nutrition (2000) vol. 71, pp. (65) Prior Publication Data 1682S-1687S. Schaffer, Amanda, “Someday, there will be a fat pill.” (Apr. 26, US 201O/OO61967 A1 Mar. 11, 2010 2005), Available: www.slate.com/id/2117332. (Accessed: Jan. 1, 2006). Stanely, Sarah, et al., “Hormonal regulation of food intake.” Physiol Related U.S. Application Data Rey (2005) vol. 85, pp. 1131-1158. (60) Provisional application No. 60/762.491, filed on Jan. Stock, Sue, et al., “Ghrelin, PYY. GP and hunger responses to a mixed meal in anorexic, obese and control female adolescents.” Journal of 27, 2006. Clinical Endocrinology & Metabolism (2005) pp. 1-36. Tovar, Sulay A., et al., “Regulation of peptide YY levels by age (51) Int. Cl. hormonal, and nutritional status.” Obesity Research (2004) vol. 12:12 A6 IK35/74 (2006.01) pp. 1944-1950. (52) U.S. Cl. .................... 424/93.45; 435/252.9; 435/29: 530/350 (Continued) (58) Field of Classification Search ........................ None Primary Examiner — Hope Robinson See application file for complete search history. (74) Attorney, Agent, or Firm — Steptoe & Johnson LLP (56) References Cited (57) ABSTRACT Use of at least one strain of a microorganism and/or a metabo lite thereof in the manufacture of a support for administration U.S. PATENT DOCUMENTS to a Subject for modulating satiety signalling, wherein the 5,578.302 A 11/1996 Brassart et al. Support is a pharmaceutically acceptable Support or a food 2005, 0186.189 A1 8, 2005 Hsu et al. product. Suitably, the at least one strain of a microorganism and/or a metabolite thereof may be administered to the sub FOREIGN PATENT DOCUMENTS ject for the treatment and/or prevention of excess weight JP 2001-292728 10, 2001 WO WOO1 (88.095 A1 11, 2001 and/or a disease caused by excess weight. Likewise, the at WO WOO2,381.65 A1 5, 2002 least one strain of a microorganism and/or a metabolite WO WO 2004/O14403 A1 2, 2004 thereof is administered to the subject for the treatment and/or WO WO 2006/025643 A1 3, 2006 prevention of obesity and/or a caused by obesity. Preferably, WO WO 2006/052135 A2 5, 2006 the microorganism is a probiotic microorganism. Suitably the WO WO 2007/043933 A1 4/2007 microorganism may be a lactic acid bacterium. Li one embodiment the microorganism is a strain of Lactobacillus OTHER PUBLICATIONS spp. and/or Bifidobacterium spp., for example a strain of Ali, Ali, A., et al., “Effects of probiotics and isoflavones on metabolic Lactobacillus acidophilus, L. curvatus, L. Salivarius and/or parameters in a genetic model of obesity and diabetes.” FASEBJour B. lactis. nal (2002) vol. 16:5. 15 Claims, 7 Drawing Sheets US 8,257,695 B2 Page 2 OTHER PUBLICATIONS Wu, Ming-Shiang, et al., “A case-control study of association of Adams, V.C., et al., “Ghrelin and cannabinoids increase food intake Helicobacter pylori infection with morbid obesity in Taiwan.” Arch via stimulation of hypothalamic amp-activated protein kinase Intern Med. (2005) vol. 165, pp. 1552-1555. (AMPK).” Endocrine (2005) (Abstract only). Vickers, S.P. et al., "Cannabinoids and the regulation of ingestive Yadav, Hariom, et al., “Antidiaberic effects of probiotic dahi contain behavior.” Current Drug Targets (2005) vol. 6:2, pp. 215-223 ing Lactobacillus acidophilus and Lactobacillus casei in high fruc (Abstract Only). tose fed rats.” Nutrition (2007) vol. 23:1, pp. 62-68. U.S. Patent Sep. 4, 2012 Sheet 1 of 7 US 8,257,695 B2 FIGURE 1 Gene expression after exposure with L.acidophilus, of Peptide YY (PYY), (meaniSE) Fold differencel100ng total RNA : L.acidophilus-treated U.S. Patent Sep. 4, 2012 Sheet 2 of 7 US 8,257,695 B2 FIGURE 2 Effect of L. acidophilus conditioned culture medium on PYY expression in Caco-2 at different glucose concentrations (mean SS 0.5 M SE) Glucose 5 mM Glucose ##### With 10% L. acid hilu ditioned ############!E ture broth U.S. Patent Sep. 4, 2012 Sheet 3 of 7 US 8,257,695 B2 FIGURE 3 Effect of various probiotic strains on Peptide YY expression, (mean tSE) | , U.S. Patent Medium Ctrl U.S. Patent Sep. 4, 2012 Sheet 5 of 7 US 8,257,695 B2 FIGURE 5 2500 N 2O O O NN 1500 1000 N 500 O SS š 2 Medium Ctrl complex lipid 10% MRS 10% MRS + 10% 10% micelles complex lipid acidophilus acidophilus micelles metabolites metabolites + complex lipid micelles U.S. Patent Sep. 4, 2012 Sheet 6 of 7 US 8,257,695 B2 FIGURE 6 PYY, time series, meaniSE s s S 2O O O s s 3e. s 9 1 O O O 2 E 5 O O Enterostim 0.1%metabolites 1%metabolites 10%metabolites LA NCFM bact U.S. Patent Sep. 4, 2012 Sheet 7 Of 7 US 8,257,695 B2 FIGURE 7 PYY expression, L. curvatus 853, MeaniSE s s 9 s s E. C. s s s 4. E Enterostin L. Curvatus 853 US 8,257,695 B2 1. 2 METHOD FOR MODULATING SATIETY Surprisingly, obestatin is encoded by the same gene that SIGNALING WITH SPECIFIC STRANS OF also encodes ghrelin, a peptide hormone that increases appe LACTOBACILLUS ACIDOPHILUS AND tite. Ghrelin and obestatin are both derived from a prohor BACILLUS mone produced by the same gene and are divided by post translational processing. The purpose of this mechanism CLAIM OF PRIORITY remains unclear, however it explains earlier findings, namely that removing the ghrelingene from mice did not significantly This application claims priority under 35 USC 371 to Inter reduce their appetite. national Application No. PCT/IB2007/001186, filed on Jan. Obestatin has been considered for use as a drug against 26, 2007, which claims priority to U.S. Provisional Applica 10 obesity, however it would have to be delivered as a nasal spray tion Ser. No. 60/762,491, filed Jan. 27, 2006, each of which is or by injection, as the peptide is destroyed by gastric juices. incorporated by reference in its entirety. There are currently few treatments for obesity. Of the two FIELD OF INVENTION drugs approved for use in the US Roche's XenicalTM (which 15 blocks the digestion offat) is relatively effective at promoting The present invention relates to Satiety of appetite. weight loss, but has some unpleasant side-effects. The other In one embodiment the present invention relates to the use drug approved for use in the US is Abbot Laboratories of at least one strain of a microorganism, preferably a lactic MeridiaTM, which has allegedly been proven to be not par acid bacterium, preferably a probiotic bacterium, Such as ticularly effective (Schaffer A. www.slate.com/id/2117332, Lactobacillus spp. (for example L. acidophilus, L. Salivarius 26 Apr. 2005). and/or L. curvatus) and/or Bifidobacterium spp. (Such as B. One experimental drug currently in trials is RimonabantTM lactis), to prepare a Support administered to humans or ani (a cannabinoid receptor antagonist) which allegedly stems mals for modulating Satiety signalling, preferably inducing cravings in humans, thus reducing obese patients appetites.
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