Cellular factors involved in the cytotoxicity of staphylococcus aureus leukocidins: novel therapeutic targets

Abstract

The present invention relates to methods for treating and preventing Staphylococcus aureus infection and/or a condition resulting from a S. aureus infection in a subject that involves administering compositions that inhibit S. aureus interaction with CXCR1/CXCR2 and DARC cellular receptors. The present invention further relates to novel compositions for carrying out these and other methods.

What is claimed: 
     
         1 . A method of preventing or treating  Staphylococcus aureus  infection and/or a condition resulting from a  S. aureus  infection in a subject comprising:
 selecting a subject having or at risk of having  S. aureus  infection and   administering, to the selected subject, a composition that inhibits  S. aureus  interaction with CXCR1 and CXCR2, under conditions effective to prevent or treat  S. aureus  infection and/or a condition resulting from a  S. aureus  infection in the subject.   
     
     
         2 . The method of  claim 1 , wherein the  S. aureus  infection is a methicillin-resistant  S. aureus  (MRSA) infection or a methicillin sensitive  S. aureus  (MSSA) infection. 
     
     
         3 . The method of  claim 1 , wherein the composition comprises an agent that inhibits both CXCR1 and CXCR2. 
     
     
         4 . The method of  claim 3 , wherein the agent is a protein or peptide inhibitor of CXCR1 and CXCR2. 
     
     
         5 . The method of  claim 4 , wherein the protein or peptide inhibitor is derived from CXC chemokine ligand 8 (CXCL8) and is selected from the group consisting of CXCL8(3-74)K11R/G31P (SEQ ID NO:1), CXCL8 (3-74) K11R/G31P/P32G (SEQ ID NO:2) and CXCL8 (3-74) K11R/T12S/H13F/G31P (SEQ ID NO:3). 
     
     
         6 . The method of  claim 4 , wherein the protein or peptide inhibitor is a recombinant peptide comprising a  S. aureus  CXCR1/CXCR2 receptor binding domain sequence. 
     
     
         7 . The method of  claim 6 , wherein the recombinant peptide comprises the amino acid sequence corresponding to amino acid residues 182-196 of SEQ ID NO:4. 
     
     
         8 . The method of  claim 6 , wherein the recombinant peptide comprises the amino acid sequence corresponding to amino acid residues 180-192 of SEQ ID NO:6. 
     
     
         9 . The method of  claim 3 , wherein the agent is a small molecule inhibitor of CXCR1 and CXCR2. 
     
     
         10 . The method of  claim 9 , wherein the small molecule inhibitor is selected from the group consisting of 2-hydroxy-N,N-dimethyl-3-[[2-[[1(R)-(5-methyl-2-furanyl)propyl]amino]-3,4-dioxo-1-cyclobuten-1-yl]amino]benzamide (SCH-527123), N-(2-hydroxy-3-dimethylsulfonylamido-4-chlorophenyl)-N′-(2-bromophenyl)-N″-cyanoguanidine (SCH468477), SCH-479833, and derivatives thereof. 
     
     
         11 . The method of  claim 9 , wherein the small molecule inhibitor is selected from the group consisting of R(−)-2-[(4-isobutylphenyl)propionyl]-methanesulfonamide (Reparixin), R(−)-2-[(4′-trifluoromethanesulfonyloxy)phenyl]propionyl-methanesulfonamide (Meraxin), 4-[(1R)-2-amino-1-methyl-2-oxoethyl]phenyl trifluoromethane sulfonate (DF 2162), and derivatives thereof. 
     
     
         12 . The method of  claim 1 , wherein the composition comprises a CXCR2 inhibitor selected from the group consisting of SB-656933, N-(2-hydroxy-3-sulfamyl-4-chlorophenyl)-N′-(2,3 dichlorophenyl)urea (SB-332235), N-(2-hydroxy-4-nitrophenyl)-N′-(2-bromophenyl)urea (SB 225002) and derivatives thereof. 
     
     
         13 . The method of  claim 1 , wherein the composition comprises a CXCR1 blocking antibody or antibody binding portion thereof, a CXCR2 blocking antibody or antibody binding portion thereof, or a combination thereof. 
     
     
         14 . The method of  claim 1 , wherein the composition comprises an antibody, or antibody binding portion thereof, that binds to an epitope of  S. aureus  Leukocidin E (LukE) comprising the amino acid sequence corresponding to amino acid residues 182-196 of SEQ ID NO:4. 
     
     
         15 . The method of  claim 1 , wherein the composition comprises an antibody, or antibody binding portion thereof, that binds to an epitope of  S. aureus  γ-Hemolysin A (HlgA) comprising the amino acid sequence corresponding to amino acid residues 180-192 of SEQ ID NO:6. 
     
     
         16 . The method of  claim 1  further comprising
 administering, in combination with said composition, an agent selected from the group consisting of an anti-infective agent, an antibiotic agent, and an antimicrobial agent. 
 
     
     
         17 . The method of  claim 1 , wherein a condition resulting from  S. aureus  infection is treated or prevented, said condition being selected from the group consisting of skin wounds and infections, tissue abscesses, folliculitis, osteomyelitis, pneumonia, scalded skin syndrome, septicemia, septic arthritis, myocarditis, endocarditis, and toxic shock syndrome. 
     
     
         18 . The method of  claim 1  further comprising:
 repeating said administering. 
 
     
     
         19 . The method of  claim 1 , wherein the subject is an infant, a juvenile, or an adult. 
     
     
         20 . The method of  claim 1 , wherein the subject is an immuno-compromised infant, juvenile, or adult. 
     
     
         21 . The method of  claim 1 , wherein the  S. aureus  infection and/or the condition resulting from a  S. aureus  infection is prevented. 
     
     
         22 . The method of  claim 1 , wherein the  S. aureus  infection and/or the condition resulting from a  S. aureus  infection is treated. 
     
     
         23 . The method of  claim 1 , wherein CXCR1 and CXCR2 interaction with LukE is inhibited. 
     
     
         24 . The method of  claim 1 , wherein CXCR1 and CXCR2 interaction with HlgA is inhibited. 
     
     
         25 . A method of preventing or treating  Staphylococcus aureus  infection and/or a condition resulting from a  S. aureus  infection in a subject comprising:
 selecting a subject having or at risk of having  S. aureus  infection and   administering, to the selected subject, a composition that inhibits  S. aureus  interaction with Duffy antigen receptor for chemokines (DARC), under conditions effective to prevent or treat  S. aureus  infection and/or a condition resulting from a  S. aureus  infection in the subject.   
     
     
         26 . The method of  claim 25 , wherein the  S. aureus  infection is a methicillin-resistant  S. aureus  (MRSA) infection or a methicillin sensitive  S. aureus  (MSSA) infection. 
     
     
         27 . The method of  claim 25 , wherein the composition comprises a DARC inhibitor. 
     
     
         28 . The method of  claim 27 , wherein the DARC inhibitor is a DARC blocking antibody. 
     
     
         29 . The method of  claim 25  further comprising
 administering, in combination with said composition, an agent selected from the group consisting of an anti-infective agent, an antibiotic agent, and an antimicrobial agent. 
 
     
     
         30 . The method of  claim 25 , wherein a condition resulting from  S. aureus  infection is treated or prevented, said condition being selected from the group consisting of skin wounds and infections, tissue abscesses, folliculitis, osteomyelitis, pneumonia, scalded skin syndrome, septicemia, septic arthritis, myocarditis, endocarditis, and toxic shock syndrome. 
     
     
         31 . The method of  claim 25  further comprising:
 repeating said administering. 
 
     
     
         32 . The method of  claim 25 , wherein the subject is an infant, a juvenile, or an adult. 
     
     
         33 . The method of  claim 25 , wherein the subject is an immuno-compromised infant, juvenile, or adult. 
     
     
         34 . The method of  claim 25 , wherein the  S. aureus  infection and/or the condition resulting from a  S. aureus  infection is prevented. 
     
     
         35 . The method of  claim 25 , wherein the  S. aureus  infection and/or the condition resulting from a  S. aureus  infection is treated. 
     
     
         36 . The method of  claim 25 , wherein DARC interaction with LukE is inhibited. 
     
     
         37 . The method of  claim 25 , wherein DARC interaction with HlgA is inhibited. 
     
     
         38 . A method of treating a subject having a  S. aureus  infection comprising:
 obtaining a sample from the subject having  S. aureus  infection;   quantifying expression levels of CXCR1, CXCR2, DARC or a combination thereof in the sample; and   administering a treatment for the subject based on said quantifying.   
     
     
         39 . The method of  claim 38 , wherein said quantifying comprises:
 measuring CXCR1, CXCR2, and/or DARC protein expression in the sample from the subject.   
     
     
         40 . The method of  claim 38 , wherein said quantifying comprises:
 measuring CXCR1, CXCR2, DARC mRNA expression in the sample from the subject.   
     
     
         41 . The method of  claim 38  further comprising:
 comparing the quantified expression levels of CXCR1, CXCR2, and/or DARC in the sample from the subject to reference expression levels of CXCR1, CXCR2, and/or DARC, wherein said suitable treatment is determined based on said comparing. 
 
     
     
         42 . An isolated LukE antibody, or antibody binding fragment thereof, wherein said antibody or binding fragment thereof, binds an epitope corresponding to amino acid residue 182-196 of SEQ ID NO:4. 
     
     
         43 . The isolated antibody of  claim 42 , wherein the antibody binding fragment comprises a single variable V H  domain, single variable V L  domain, a Fab fragment, or a single-chain antibody fragment. 
     
     
         44 . An isolated HlgA antibody, or antibody binding fragment thereof, wherein said antibody or binding fragment thereof, binds an epitope corresponding to amino acid residue 180-192 of SEQ ID NO:6. 
     
     
         45 . The isolated antibody of  claim 44 , wherein the antibody binding comprises a single variable V H  domain, single variable V L  domain, a Fab fragment, or a single-chain antibody fragment. 
     
     
         46 . A composition comprising
 an isolated LukE protein or polypeptide thereof having a non-functional CXCR1/CXCR2 binding domain and   a pharmaceutically acceptable carrier   
     
     
         47 . The composition of  claim 46 , wherein the isolated LukE polypeptide comprises amino acid residues 20-263 of SEQ ID NO:4, wherein one or more amino acid residues corresponding to amino acid residues 182-196 of SEQ ID NO:4 is substituted or deleted. 
     
     
         48 . The composition of  claim 47 , wherein one or more amino acids selected from the group consisting of P184, G186, P187, and G189 is substituted or deleted. 
     
     
         49 . The composition of  claim 47 , wherein the isolated LukE protein comprises the amino acid sequence of SEQ ID NO:4. 
     
     
         50 . The composition of  claim 47 , wherein the isolated LukE protein comprises the amino acid sequence of SEQ ID NO:8 or SEQ ID NO:9. 
     
     
         51 . The composition of  claim 57 , wherein the isolated LukE polypeptide comprises the amino acid sequence of SEQ ID NO:10 or SEQ ID NO;11. 
     
     
         52 . The composition of  claim 46  further comprising
 an isolated Leukocidin D (LukD) protein or polypeptide thereof. 
 
     
     
         53 . The composition of  claim 52 , wherein the composition comprises an isolated LukD protein comprising the amino acid sequence of SEQ ID NO:12 or an amino acid sequence having at least 70% sequence similarity to SEQ ID NO:12. 
     
     
         54 . The composition of  claim 52 , wherein the composition comprises an isolated LukD polypeptide comprising amino acid residues 20-281 of SEQ ID NO:12. 
     
     
         55 . A composition comprising
 an isolated HlgA protein or polypeptide thereof having a non-functional CXCR1/CXCR2 binding domain and   a pharmaceutically acceptable carrier.   
     
     
         56 . The composition of  claim 55 , wherein the isolated HlgA protein comprises amino acid residues 20-258 of SEQ ID NO:6, wherein one or more amino acid residues corresponding to amino acid residues 180-192 of SEQ ID NO:6 is substituted or deleted. 
     
     
         57 . The composition of  claim 56 , wherein one or more amino acids selected from the group consisting of P182, G184, and P185 is substituted or deleted. 
     
     
         58 . The composition of  claim 56 , wherein the isolated HlgA protein comprises the amino acid sequence of SEQ ID NO:6. 
     
     
         59 . The composition of  claim 56 , wherein the isolated HlgA protein comprises the amino acid sequence of SEQ ID NO:13, SEQ ID NO:14, or SEQ ID NO:15. 
     
     
         60 . The composition of  claim 55  further comprising
 an isolated γ-Hemolysin B (HlgB) protein or polypeptide thereof. 
 
     
     
         61 . The composition of  claim 60 , wherein the composition comprises an isolated HlgB protein comprising the amino acid sequence of SEQ ID NO:16 or an amino acid sequence having at least 70% sequence similarity to SEQ ID NO:16. 
     
     
         62 . A method of preventing or treating Human Immunodeficiency Virus (HIV) infection in a subject, said method comprising:
 selecting a subject having or at risk of having HIV infection and   administering, to the selected subject, the composition of  claim 46  or  claim 52  under conditions effective to prevent or treat HIV in the subject.   
     
     
         63 . The method of  claim 62  further comprising:
 administering one or more antiviral or other agents useful in the treatment of HIV in combination with the composition comprising the isolated LukE protein, or polypeptide thereof, and the isolated LukD protein, or polypeptide thereof. 
 
     
     
         64 . The method of  claim 63 , wherein the antiviral agent is selected from the group consisting of a nucleoside reverse transcriptase inhibitor, a non-nucleoside reverse transcriptase inhibitor, and a protease inhibitor. 
     
     
         65 . The method of  claim 63 , wherein the antiviral agent is selected from the group consisting of zidovudine, lamivudine, zalcitabine, didanosine, stavudine, abacavir, adefovir dipivoxil, lobucavir, BC H-10652, emitricitabine, beta-L-FD4, DAPD, lodenosine, nevirapine, delaviridine, efavirenz, PNU-142721, AG-1549, MKC-442, (+)-calanolide A and B, saquinavir, indinavir, ritonavir, nelfinavir, lasinavir, DMP-450, BMS-2322623, ABT-378, amprenavir, hydroxyurea, ribavirin, IL-2, IL-12, pentafuside, Yissum No. 1 1607, and AG-1549. 
     
     
         66 . The composition of  claim 46 ,  claim 52 ,  claim 55 , or  claim 60  further comprising:
 one or more additional agents selected from the group consisting of a lubricant, an antimicrobial agent, a humectant, an emulsifier, and a mixture of two or more thereof. 
 
     
     
         67 . The composition of  claim 66 , wherein the composition comprises a lubricant, said lubricant being selected from the group consisting of cetyl esters wax, hydrogenated vegetable oil, magnesium stearate, methyl stearate, mineral oil, polyoxyethylene-polyoxypropylene copolymer, polyethylene glycol, polyvinyl alcohol, sodium lauryl sulfate or white wax, and a mixture of two or more thereof. 
     
     
         68 . The composition of  claim 66 , wherein the composition comprises an antimicrobial agent, said antimicrobial agent being selected from the group consisting of propylene glycol, methyl paraben, propyl paraben, and a mixture of two or more thereof. 
     
     
         69 . The composition of  claim 66 , wherein the composition comprises an antioxidant, said antioxidant being selected from the group consisting of butylated hydroxyanisole, butylated hydroxytoluene, edetate disodium, and a mixture of two or more thereof. 
     
     
         70 . The composition of  claim 66 , wherein the composition comprises a humectant, said humectant being selected from the group consisting of ethylene glycol, glycerin, sorbitol, or a mixture of two or more thereof. 
     
     
         71 . The composition of  claim 66 , wherein the composition comprises an emulsifier, said emulsifier being selected from the group consisting of carbomer, polyoxyethylene-10-stearyl ether, polyoxyethylene-20-stearyl ether, cetostearyl alcohol, cetyl alcohol, cholesterol, diglycol stearate, glyceryl monostearate, glyceryl stearate, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, lanolin, polyoxyethylene lauryl ether, methyl cellulose, polyoxyethylene stearate, polysorbate, propylene glycol monostearate, sorbitan esters, stearic acid, and a mixture of two or more thereof. 
     
     
         72 . The composition of  claim 66 , wherein said composition is formulated for topical administration. 
     
     
         73 . The composition of  claim 66 , wherein said composition is formulated for vaginal and/or rectal administration. 
     
     
         74 . The composition of  claim 66 , wherein said composition is formulated as an oral rinse. 
     
     
         75 . A method of preventing HIV infection in a subject, said method comprising:
 selecting a subject at risk of being exposed to HIV infection;   providing the composition of  claim 46 ,  claim 52 , or  claim 66 , wherein the composition comprises an isolated LukE protein or polypeptide thereof having a non-functional CXCR1/CXCR2 binding domain; and   contacting tissue of the subject with the composition under conditions effective to inhibit HIV infectivity of cells in the tissue, thereby preventing HIV infection in the subject.   
     
     
         76 . A method of treating an inflammatory condition in a subject comprising:
 selecting a subject having an inflammatory condition and   administering, to the selected subject, the composition of  claim 46  or  claim 52  under conditions effective to treat the inflammatory condition.   
     
     
         77 . The method of  claim 76 , wherein the inflammatory condition is an acute inflammatory condition. 
     
     
         78 . The method of  claim 77 , wherein the acute inflammatory condition is localized. 
     
     
         79 . The method of  claim 77 , wherein the acute inflammatory condition is an infected wound in the skin or soft tissue. 
     
     
         80 . The method of  claim 76 , wherein the inflammatory condition is selected from the group consisting of rheumatoid arthritis, Crohn's disease, atherosclerosis, psoriasis, ulcerative colitis, psoriatic arthritis, multiple sclerosis, lupus, type I diabetes, primary biliary cirrhosis, inflammatory bowel disease, tuberculosis, skin wounds and infections, tissue abscesses, folliculitis, osteomyelitis, pneumonia, scalded skin syndrome, septicemia, septic arthritis, myocarditis, endocarditis, toxic shock syndrome, allergic contact dermatitis, acute hypersensitivity, and acute neurological inflammatory injury. 
     
     
         81 . A method of preventing or treating graft-versus-host-disease (GVHD) in a subject, said method comprising:
 selecting a subject having or at risk of having GVHD and   administering, to the selected subject, the composition of  claim 46  or  claim 52  in an amount effective to prevent or treat graft-versus-host-disease (GVHD) in the subject.   
     
     
         82 . A method of preventing or treating cancer in a subject, said method comprising:
 selecting a subject having or at risk of having cancer and   administering, to the selected subject, the composition of  claim 46  or  claim 52  in an amount effective to prevent or treat cancer in the subject.   
     
     
         83 . The method of  claim 82 , wherein the cancer is selected from the group consisting of breast cancer, prostate cancer, liver cancer, and lung cancer. 
     
     
         84 . The method of  claim 82 , wherein the cancer is metastatic cancer.