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WO 2009/111838 Al (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date 17 September 2009 (17.09.2009) WO 2009/111838 Al (51) International Patent Classification: [AU/AU]; 6 Doidge Street, Bundoora, Victoria 3083 C07K 14/47 (2006.01) C07K 19/00 (2006.01) (AU). A61P 1/00 (2006.01) Cl 2N 15/861 (2006.01) (74) Agents: OLIVE, Mark et al; F B RICE & CO, Level 23, A61P 31/04 (2006.01) A23C 19/091 (2006.01) 44 Market Street, Sydney, New South Wales 2000 (AU). C12N 15/85 (2006.01) A61P 21/16 (2006.01) AOlK 61/021 (2006.01) C12N 15/63 (2006.01) (81) Designated States (unless otherwise indicated, for every A61P 11/00 (2006.01) Cl 2N 15/869 (2006.01) kind of national protection available): AE, AG, AL, AM, A61P 31/10 (2006.01) A61K 38/11 (2006.01) AO, AT, AU, AZ, BA, BB, BG, BH, BR, BW, BY, BZ, C12N 15/86 (2006.01) A61P 31/00 (2006.01) CA, CH, CN, CO, CR, CU, CZ, DE, DK, DM, DO, DZ, A23C 3/00 (2006.01) C12N 15/82 (2006.01) EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, A61P 15/14 (2006.01) HR, HU, ID, IL, IN, IS, JP, KE, KG, KM, KN, KP, KR, KZ, LA, LC, LK, LR, LS, LT, LU, LY, MA, MD, ME, (21) International Application Number: MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, PCT/AU2009/000301 NZ, OM, PG, PH, PL, PT, RO, RS, RU, SC, SD, SE, SG, (22) International Filing Date: SK, SL, SM, ST, SV, SY, TJ, TM, TN, TR, TT, TZ, UA, 13 March 2009 (13.03.2009) UG, US, UZ, VC, VN, ZA, ZM, ZW. (25) Filing Language: English (84) Designated States (unless otherwise indicated, for every kind of regional protection available): ARIPO (BW, GH, (26) Publication Language: English GM, KE, LS, MW, MZ, NA, SD, SL, SZ, TZ, UG, ZM, (30) Priority Data: ZW), Eurasian (AM, AZ, BY, KG, KZ, MD, RU, TJ, 2008901249 13 March 2008 (13.03.2008) AU TM), European (AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, (71) Applicant (for all designated States except US): AGRI¬ MC, MK, MT, NL, NO, PL, PT, RO, SE, SI, SK, TR), CULTURE VICTORIA SERVICES PTY LIMITED OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, ML, [AU/AU]; 475 Mickleham Road, Attwood, Victoria 3049 MR, NE, SN, TD, TG). (AU). Published: (72) Inventors; and (75) Inventors/Applicants (for US only): COCKS, Benjamin — with international search report (Art. 21(3)) [AU/AU]; 293 Banyule Road, Viewbank, Victoria 3084 — with sequence listing part of description (Rule 5.2(a)) (AU). SPANGENBERG, German [AU/AU]; 56 Arthur Street, Bundoora, Victoria 3083 (AU). WANG, Jianghui (54) Title: METHOD OF TREATMENT USING ANTIMICROBIAL COMPOSITION (57) Abstract: The present invention provides peptides and analogs and derivatives thereof having antimicrobial activity at least against Streptococcus uberis for the treatment of a range of infectious disease including mastitis, otitis externa, clostridial intesti- nal disease and respiratory disease. Method of treatment using; antimicrobial composition Related applications This application claims priority from Australian Patent Application No. 2008901249 filed March 13, 2008, the contents of which are incorporated herein in their entirety. Field of the invention The present invention relates to antibacterial peptide reagents and methods employing same for the treatment of microbial disease(s), in particular microbial disease(s) mediated in part of whole by one or more bacteria and/or fungi. Background of the invention Human and animal health are valuable commercial sectors and bacterial and fungal pathogenic infections in humans, livestock and domestic pets represent a high cost to these sectors in terms of lost productivity and existing treatment costs. Many bacterial and fungal pathogens of diseases in humans, livestock animals and domestic pets are also recalcitrant to treatment with existing antibiotics, further exacerbating these adverse consequences of infection. For example, the economic value of the dairy industry worldwide is significant. For example, the International Dairy Foods Association estimates that sales of cow milk in USA alone in 2006 was USD 23.9 billion. This value will be significantly increased when considered on a worldwide scale, and expanded to include all dairy products, e.g., cheese and butter, and to include sales of products from all major animal producers of dairy products, e.g., goats, sheep, camels and buffalo. The pharmaceutical and biotechnology industries have also developed an interest in dairy mammals as suitable bioreactors for the production of biological agents, particularly peptides and proteins. In this respect, the combination of large daily protein output, post-translational processing capabilities, ease of access to recombinant protein by milking and low capital cost of production plants, i.e., farms compared to high volume industrial fermenters makes dairy mammals excellent candidates for the production of recombinant peptides and proteins (Echelard, Curr. Opin. Biotechnol, 7: 536-540, 1996). Mastitis is currently the most economically important disease of dairy mammals (Pyδrala Reprod. Dom Anim., 37: 211-216, 2002 and Bergonier et al, 34: 689-716, 2003). The annual costs of mastitis in dairy cattle alone is estimated to be about 10% of the total sales of farm milk, i.e., about USD 2 billion in USA alone (Radostits et Veterinary Medicine: A Textbook of the Diseases of Cattle, Sheep, Pigs, Goats and Horses, Ninth Ed., Elsevier Health Sciences, 2000). In this respect, the costs of mastitis extend beyond treatment and prevention costs and include losses in milk production, labor costs and loss of animals due to culling. Other effects of mastitis include the impact of agricultural use of antibiotics that are used to treat mastitis on the development of antibiotic resistant human pathogens (Smith et al., Proc. Natl. Acad. ScL, USA 99: 6434-6439, 2002) and the effect of residues of these antibiotics on human health (Clement Anim. Pharm., 407: 22-23, 1998). Moreover, the limited milk production resulting from mastitis limits the utility of mammals as bioreactors for producing pharmaceutical agents. Mastitis is an inflammatory reaction of the mammary gland, usually to microbial infection. This condition is characterized by an influx of somatic cells, primarily polymorphonuclear neutrophils (PMN), into the mammary gland, and by an increase in milk protease content (Verdi et al, J. Dairy ScI, 70: 230-242, 1987). Mastitis is generally classified into clinical infections and non-clinical infections. Clinical infections are diagnosed by red, swollen appearance of a mammary gland and flakes or clots (protein aggregates) in the milk. Sub-clinical infections show no obvious symptoms. The majority of cases of mastitis are caused by one or more of Staphylococcus aureus, Streptococcus dysgalactiae, Streptococcus agalactiae, Streptococcus uberis or Escherichia coli. In this respect, S. aureus, S. dysgalactiae and S. agalactiae have a contagious route of transmission, whereas S. uberis and E. coli are environmental pathogens (Kerr and Wellnitz, J. Anim. ScI, 81: 38-47, 2003). Susceptibility of a mammal to an intra-mammary infection leading to mastitis dramatically increases during early involution and during the periparturient period (Nickerson J. Dairy ScL, 72: 1665-1678, 1989; and Oliver and Sordillo, J. Dairy ScL, 71: 2584-2606, 1988). These infections are often associated with clinical mastitis during early lactation, and can have a marked detrimental effect on subsequent milk yield and/or quality. Susceptibility to mastitis is also high during the prepartum period in first-lactation bovine heifers (Nickerson et al, J. Dairy ScL, 78: 1607-1618, 1995). These infections are associated with a decrease in alveolar epithelial and luminal area and an increase in connective tissue in the mammary gland, potentially causing a life¬ long reduction in milk yield in the infected mammal. The incidence of contagious mastitis has declined over the last thirty years as a result of a five point control plan that recommends use of correctly maintained milking equipment, post-milking teat disinfection, both therapeutic and prophylactic use of antibiotics and culling of persistently infected animals (Bramley and Dodd, J. Dairy Res., 51: 481-512, 1984). Notwithstanding that implementation of this plan has almost eliminated S. dysgalactiae and S. agalactiae from many herds, as discussed supra the use of antibiotics is both expensive and may have a detrimental effect on human health. Moreover, . aureus, which accounts for 15% to 30% of contagious infections has proven to be resistant to traditional management approaches. In this respect, the cure rate of treatment of S. aureus infection is often less than 15%. This reduced cure rate is attributed in part to antibiotic resistant strains of S. aureus, and to incomplete penetration of the antibiotics through a mammary gland thereby permitting bacteria to survive within the gland. Moreover, S. aureus is able to survive within mammary gland epithelial cells, within which antibiotic concentration is insufficient to cause bacterial cell death (Craven and Anderson J. Dairy Res., 51: 513-523, 1984, and Yancey et al, Eur. J. Clin. Microbiol. Infect. Dis., 10: 107-1 13, 2991). Antibiotic treatment is used to treat mastitis caused by environmental pathogens, e.g., S. uberis, however these pathogens are often resistant to conventional antibiotics. Moreover, recurrence of infection from environmental reservoirs, e.g., within dairy barns, is a continuing problem (Kerr and Wellnitz, supra). Current therapies for mastitis make use of conventional antibiotics, e.g., β-lactams including penicillins and cephalosporins. Notwithstanding that these antibiotics may be effective in the treatment of some pathogens that cause mastitis, some bacterial pathogens are resistant to these compounds. There is also a risk that ongoing use of these compounds may contribute to emergence of antibiotic resistant human pathogens (Smith et al, Proc.
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