US 20090104204A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2009/0104204 A1 Throsby et al. (43) Pub. Date: Apr. 23, 2009 (54) HUMAN BINDING MOLECULES HAVING (30) Foreign Application Priority Data KILLING ACTIVITY AGAINST STAPHYLOCOCC AND USES THEREOF Nov. 16, 2006 (EP) .................................. O61242319 Mar. 6, 2007 (EP) .................................. O7103.584.4 (76) Inventors: Mark Throsby, Utrach (NL); Publication Classification Cecilia A.W. Geuijen, Moerkapelle (51) Int. Cl Nels Adriaan De Kruif, A 6LX 39/395 (2006.01) e Bilt (NL) C07K 6/12 (2006.01) A6IP3L/04 (2006.01) Correspondence Address: C7H 2L/04 (2006.01) TRASK BRITT CI2N 15/63 (2006.01) P.O. BOX 2.5SO CI2N 5/10 (2006.01) SALT LAKE CITY, UT 84110 (US) CI2P 2L/08 (2006.01) GOIN 33/53 (2006.01) (21) Appl. No.: 12/227,029 (52) U.S. Cl. .............. 424/142.1; 530/388.15:530/391.3; 536/23.53; 435/320.1; 435/325; 435/69.6; (22) PCT Filed: Jun. 5, 2007 435/7.1 (57) ABSTRACT (86)86) PCT No.:O PCT/EP2007/055527 The present invention provides human binding molecules S371 (c)(1) specifically binding to Staphylococci and having killing activ (2), (4) Date: Nov. 5, 2008 ity against Staphylococci, nucleic acid molecules encoding the human binding molecules, compositions comprising the human binding molecules and methods of identifying or pro Related U.S. Application Data ducing the human binding molecules. The human binding (60) Provisional application No. 60/811,477, filed on Jun. molecules can be used in the diagnosis, prophylaxis and/or 6, 2006. treatment of a condition resulting from Staphylococcus. Patent Application Publication Apr. 23, 2009 Sheet 1 of 6 US 2009/O104204 A1 75 3s 50 O O O) 25 SS O -------E.------ E.------ -E------- E H N s s Antibody concentration (ug/ml) FIG. 1 Patent Application Publication Apr. 23, 2009 Sheet 2 of 6 US 2009/O104204 A1 5 O 2 5 Antibody Concentration (ug/ml) FIG. 2 Patent Application Publication Apr. 23, 2009 Sheet 3 of 6 US 2009/O104204 A1 Antibody Concentration (ug/ml) FIG. 3 Patent Application Publication Apr. 23, 2009 Sheet 4 of 6 US 2009/O104204 A1 30 s3 20 9 O O O 10 SS t -1------t -E- -------E.------ O H H N N N Q SS Antibody Concentration (ug/ml) FIG. 4 US 2009/01 04204 A1 Apr. 23, 2009 HUMAN BINDING MOLECULES HAVING therapies or preventative measures for (multi-drug-resistant) KILLING ACTIVITY AGAINST bacterial infections is urgently needed to meet this impending STAPHYLOCOCC AND USES THEREOF healthcare crisis. 0004 Active immunization with vaccines and passive FIELD OF THE INVENTION immunization with immunoglobulins are promising alterna tives to classical small molecule therapy. A few bacterial 0001. The invention relates to medicine. In particular the diseases that once caused widespread illness, disability, and invention relates to the diagnosis, prophylaxis and/or treat death can now be prevented through the use of vaccines. The ment of infection by Staphylococci. vaccines are based on weakened (attenuated) or dead bacte ria, components of the bacterial Surface or on inactivated BACKGROUND OF THE INVENTION toxins. The immune response raised by a vaccine is mainly directed to immunogenic structures, a limited number of pro 0002 Staphylococcus is a genus of gram-positive bacteria teins or Sugar structures on the bacteria that are actively and a member of the micrococcaceae family. Staphylococci processed by the immune system. Since these immunogenic are spherical bacteria that are found primarily on the skin and structures are very specific to the organism, the vaccine needs in the mucous membranes of humans and other warm to comprise the immunogenic components of all variants of blooded animals, and aggregate into Small, grape-like the bacteria against which the vaccine should be protective. clumps. Staphylococci can be divided into two groups, i.e. As a consequence thereof, vaccines are very complex, take coagulase-positive and coagulase-negative staphylococci. long and are expensive to develop. Further complicating the Overall, there are about thirty species of staphylococci. design of vaccines is the phenomenon of antigen replace 0003 Staphylococci can cause a wide variety of diseases ment. This occurs when new strains become prevalent that in humans either through toxin production or invasion. Sta are serologically and thus antigenically distinct from those phylococcus aureus (S. aureus) has been recognized as one of strains covered by the vaccines. The immune status of the the most important and lethal human bacterial pathogens populations at risk for nosocomial infections further compli since the beginning of the previous century. Until the antibi cates vaccine design. These patients are inherently unwell otic era, more than 80% of the patients growing S. aureus and may even be immunocompromised (due to the effect of from their blood died. Through infections caused by coagul immunosuppressive drugs) resulting in delayed or insuffi lase-positive S. aureus were generally known to be poten cient immunity against the infecting pathogens. Furthermore, tially lethal, coagulase-negative staphylococci has been dis except in the case of certain elective procedures, it may not be missed as avirulent skin commensals incapable of causing possible to identify and vaccinate the at risk patients in time to human disease. However, over the past 30 years, coagulase give them Sufficient immune protection from infection. negative staphylococcal infections have emerged as one of 0005 Direct administration of therapeutic immunoglobu the major complications of medical progress. They are cur lins, also referred to as passive immunization, does not rently the pathogens most commonly isolated from infections require an immune response from the patient and therefore of indwelling foreign devices and are the leading cause of gives immediate protection. In addition, passive immuniza nosocomial (hospital-acquired) bacteremias in US hospitals. tion can be directed to bacterial structures that are not immu Staphylococcal infections are commonly treated with antimi nogenic and that are less specific to the organism. Passive crobial agents. However, the ascendancy of staphylococci as immunization against pathogenic organisms has been based pre-eminent nocosomial pathogens also has been associated on immunoglobulins derived from Sera of human or non with a major increase in the proportion of these isolates that human donors. However, blood-derived products have poten are resistant to (multiple) antimicrobial agents. Of the esti tial health risks inherently associated with these products. In mated 2 million hospital infections in the US in 2004, 70% addition, the immunoglobulins can display batch-to-batch was resistant to at least one antibiotic, thereby causing major variation and may be of limited availability in case of sudden medical and consequently economic problems. Ninety per mass exposures. Recombinantly produced antibodies do not cent of the staphylococci strains are penicillin resistant, leav have these disadvantages and thus offer an opportunity to ing only methicillin and Vancomycin to treat the majority of replace immunoglobulins derived from Sera. infections. However, with increasing numbers of reports of 0006 Murine monoclonal antibodies directed against sta methicillin-resistant Staphylococcus aureus (MRSA) chem phylococciare known in the art (see WO 03/059259 and WO ists are faced with the daunting task of generating new anti 03/059260). However, murine antibodies are limited for their biotics with novel modes of action. Despite the urgent need use in vivo due to problems associated with administration of for the development of new antibiotics, the major pharmaceu murine antibodies to humans, such as short serum halflife, an tical companies appear to have lost interest in the antibiotic inability to trigger certain human effector functions and elici market. In 2002, only 5 out of the more than 500 drugs in tation of an unwanted dramatic immune response against the phase II or phase III clinical development were new antibiot murine antibody in a human (HAMA). ics. In the last 6 years only 10 antibiotics have been registered 0007. In WO 03/059259 and WO 03/059260 the attempts and only 2 of those did not exhibit cross-reactivity with exist have been made to overcome the problems associated with the ing drugs (and thus not subject to the same patterns of drug use of fully murine antibodies in humans by preparing chi resistance). This trend has been attributed to several factors: meric antibodies. A disadvantage of these chimeric antibod the cost of new drug development and the relatively small ies is however that they still retain some murine sequences return on investment that infectious disease treatments yield and therefore still elicit an unwanted immune reaction, espe compared to drugs against hypertension, arthritis and lif cially when administered for prolonged periods. estyle drugs e.g. for impotence. Another contributing factoris 0008 WO 2004/043405 relates to polysaccharide vac the increasing difficulty in finding new targets, further driving cines for staphylococcal infections, prepared from poly up development costs. Therefore, investigation into novel N-acetylglucosamine (PNAG) surface polysaccharide from US 2009/01 04204 A1 Apr. 23, 2009 Staphylococci, and the deacetylated form thereof (dPNAG). acid residues, at least 10 contiguous amino acid residues, at WO 2004/043405 also discloses rabbit antiserum to PNAG least 15 contiguous amino acid residues, at least 20 contigu and dPNAG, coupled to Diphteria Toxoid (DTm). ous amino acid residues, at least 25 contiguous amino acid 0009. Although WO 03/059259, WO 03/059260 and WO residues, at least 30 contiguous amino acid residues, at least 20047043405 refer to human antibodies as desired molecules, 35 contiguous amino acid residues, at least 40 contiguous the antibodies actually disclosed and used therein are partly amino acid residues, at least 50 contiguous amino acid resi of murine or completely of rabbit origin, and none of these dues, at least 60 contiguous amino residues, at least 70 con documents actually discloses any human antibodies, nor tiguous amino acid residues, at least 80 contiguous amino sequences thereof.