(19) TZZ _T (11) EP 2 249 824 B1 (12) EUROPEAN PATENT SPECIFICATION (45) Date of publication and mention (51) Int Cl.: of the grant of the patent: A61K 31/23 (2006.01) A61P 31/00 (2006.01) 16.07.2014 Bulletin 2014/29 A23K 1/16 (2006.01) A23K 1/18 (2006.01) (21) Application number: 09703269.2 (86) International application number: PCT/EP2009/050770 (22) Date of filing: 23.01.2009 (87) International publication number: WO 2009/092787 (30.07.2009 Gazette 2009/31) (54) LACTYLATES FOR THE PREVENTION AND TREATMENT OF INFECTIONS CAUSED BY GRAM- POSITIVE BACTERIA IN ANIMALS LACTYLATE ZUR PRÄVENTION UND BEHANDLUNG VON INFEKTIONEN DURCH GRAMPOSITIVE BAKTERIEN BEI TIEREN LACTYLATES CONVENANT POUR LA PRÉVENTION ET LE TRAITEMENT D’INFECTIONS PAR BACTÉRIES GRAM POSITIVES CHEZ DES ANIMAUX (84) Designated Contracting States: (56) References cited: AT BE BG CH CY CZ DE DK EE ES FI FR GB GR EP-A- 1 483 975 WO-A-01/06877 HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL WO-A-2004/037177 US-A1- 2006 062 832 PT RO SE SI SK TR US-A1- 2007 010 856 (30) Priority: 25.01.2008 EP 08100911 • DATABASE WPI Week 2008 Thomson Scientific, London, GB; AN 2008-D16825 XP002523620 (43) Date of publication of application: WANG J; WANG Y: "FEED ADDITIVE ANIMAL 17.11.2010 Bulletin 2010/46 COMPRISE LIPASE EMULSION EXTRACT SUBSIDIARY MATERIAL" & CN 101 053 368 A (73) Proprietor: PURAC Biochem BV ((GUAN-N)GUANGZHOU WEITEFU BIOLOGICAL 4206 AC Gorinchem (NL) & SCI CO LTD) 17 October 2007 (2007-10-17) & CN 101 053 368 A (GUANGZHOU WEITEFU (72) Inventor: CAZEMIER, Anne BIOLOG TECHN [CN]) 17 October 2007 NL-4143 HE Leerdam (NL) (2007-10-17) (74) Representative: De Vries & Metman Remarks: Overschiestraat 180 Thefile contains technical information submitted after 1062 XK Amsterdam (NL) the application was filed and not included in this specification Note: Within nine months of the publication of the mention of the grant of the European patent in the European Patent Bulletin, any person may give notice to the European Patent Office of opposition to that patent, in accordance with the Implementing Regulations. Notice of opposition shall not be deemed to have been filed until the opposition fee has been paid. (Art. 99(1) European Patent Convention). EP 2 249 824 B1 Printed by Jouve, 75001 PARIS (FR) EP 2 249 824 B1 Description [0001] The present invention pertains to a method for preventing or treating intestinal infections caused by gram- positive bacteria in animals, to specified compositions for preventing or treating intestinal infections caused by gram- 5 positive bacteria in animals, to the use of specified compositions for preventing or treating intestinal infections caused by gram-positive bacteria in animals, and to a nutrition composition for animals comprising a specific compound in an amount effective for preventing or treating intestinal infections caused by gram-positive bacteria in animals. [0002] Gram-positive bacteria are stained dark blue or violet by gram staining, mainly due to a high amount of pepti- doglycan in their cell wall. Among the gram positive bacteria are the pathogenic bacteriaEnterococcus, Clostridium, 10 Listeria, Staphylococcus, various Bacillus species, and Streptococcus. While some of these organism are mainly of concern as food contaminants, others can cause diseases in animals. [0003] For example, Clostridia are responsible for causing a number of widely varying diseases of the intestine in animals. As it is a nearly ubiquitous bacteria readily found in soil, dust, faeces, and feed, it is extremely difficult to keep animals free from Clostridia. 15 Clostridium-related intestinal diseases may be quite severe. For example,Clostridia are involved in causing necrotic enteritis in chickens. In cattle, Clostridia-related enteritis can take the form of "sudden death syndrome", which, in practice can result in the in overnight deaths of a number of cattle. Also in other animals, Clostridia-related diseases may cause severe damage. [0004] It is known to administer antibiotics to animals to protect them from intestinal infection. It is also known to include 20 such antibiotics in animal nutrition. However, there is an increasing resistance against the use of antibiotics in animal feed, and nowadays many countries have legislation that prohibits the use of antibiotics in animal feed. Moreover, antibiotics have to be administered in very controlled amounts. [0005] Accordingly, there is therefore a need for a non-antibiotic method and composition for animal feed that will help to treat or prevent intestinal infections caused by gram-positive bacteria in animals, in particular in mammals, including 25 ruminants (e.g. cattle, sheep, goat, deer) and monogastrics (e.g. swine, horses, rabbits); in birds (e.g. poultry, turkey, pheasant, quail); in fish, including marine fish (e.g. salmon halibut, tuna), fresh water fish (e.g. trout, carp, tilapia); molluscs (e.g. oyster, mussels, clam, snail) and crustacean (e.g. crab, lobster. shrimp). The present invention may also find application in humans, and in fur animals such as mink, ermine, sabre, and foxes. [0006] Further, it may be desirable to suppress specific Gram-positive bacteria in the intestine with a view to increasing 30 the growth of animals. It is believed that this may be of interest for Lactobacillus spp. The present invention is also of interest for this application. [0007] The present invention provides a method and composition for animal feed for treating or preventing intestinal infections caused by gram-positive bacteria in animals. In accordance with the present invention, use is made of an antibacterial compound selected from lactylate in accordance with formula 1, 35 Formula 1 R2-COO-[-CH(CH3)-COO]n-R1 or a Na, K, Ca, Mg, Fe(II), Zn, NH 4, or Cu(II) salt thereof, a glycolylate of formula 2, 40 Formula 2: R2-COO-[-CH2-COO]n-R1 or a Na, K, Ca, Mg, Fe(II), Zn, NH 4, or Cu(II) salt thereof a lactate ester of formula 3, 45 Formula 3: HO-CH(CH3)-COO-R2 and/or a glycolic acid ester of formula 4, 50 Formula 4: HO-CH2-COO-R2 wherein in the above formulas R1 is selected from H, n stands for an integer with a value of 1-10, and R2 stands for a C1-C35 alkyl or alkenyl chain which may be branched or unbranched for the prevention or treatment of intestinal infections caused by gram-positive bacteria. 55 [0008] The present invention pertains to the prevention or treatment of intestinal infections by gram-positive bacteria in animals. The invention is particularly attractive for use against intestinal infections with anaerobic or facultative anaer- obic bacteria, even more in particular anaerobic bacteria. Within the group of anaerobic bacteria, it is particularly desirable to have a method for the prevention or treatment of intestinal infections by spore-forming bacteria, as these organism 2 EP 2 249 824 B1 tend to be difficult to control. The invention is of particular interest in the prevention and treatment of intestinal infectio ns by Clostridia. In one embodiment, the present invention pertains to the prevention or treatment of intestinal infections caused by Clostridium, in particular by Clostridium perfringens in poultry, in particular in chicken. 5 In another embodiment the present invention pertains to the prevention or treatment of intestinal infections caused by Clostridium, in particular by one or more of Clostridium tetanii, novyi (type B) septicum, chauvii, sordelii, hemolyticum, difficile, botulinum, in cattle. In a further embodiment the present invention pertains to the reduction of intestinal growth of Lactobacillus spp. [0009] It is noted that WO 2004/107877 describes an antimicrobial composition comprising a mixture of lactic acid or 10 a derivative thereof and an inorganic acid. The composition is described as antimicrobial in general. The use against Salmonella and Escherichia Coli is specified. While lactylates are mentioned as possible lactic acid derivatives, their use is not further elucidated. There is nothing in this reference that teaches or suggests the particular efficacy that the use of lactylates has been found to show against gram-positive bacteria in animals. [0010] It is further noted that GB115480 describes the use of acylated alpha-hydroxy carboxylic acids against bacteria 15 and fungi, for instance moulds, mildews, and yeasts. It is indicated that the compound can be used for consumption by or application to humans or other animals, but this is never elucidated. There is nothing in this reference that teaches or suggests the particular efficacy that the use of lactylates has been found to show against gram-positive bacteria. [0011] Furthermore, Wang and Wang (2007, AN 2008-D116825 XP002523620) discloses the use of calcium stearyl lactylate as emulsifier in feed additive for animal diarrhea resulting from fat indigestion. 20 WO01/06877A discloses a method for reducing the mumber of gram positive bacteria in food and non food products comprising the step of treating the products with a bacteriostatically or bactericidally effective amount of a composition comprising sodium stearoyl-2 lactylate. WO2004/037177A discloses shaving gel composition with antimicrobial agents comprising also sodium isostearyl lactylate. US2007/010856A1 discloses multifilament prepared by permeating the suture with an antimicrobial solution comprising lactylates. 25 [0012] In the present invention, use may be made of an an antibacterial compound selected from one or more of a lactylate in accordance with formula 1, or a Na, K, Ca, Mg, Fe(II), Zn, NH 4, or Cu(II) salt thereof, a glycolylate of formula 2, or a Na, K, Ca, Mg, Fe(II), Zn, NH4, or Cu(II) salt thereof, a lactate ester of formula 3, and/or a glycolic acid ester of formula 4. [0013] The use of a lactylate of formula 1 or a salt thereof has been found to be preferred.