US 20120058935A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2012/0058935 A1 Moir et al. (43) Pub. Date: Mar. 8, 2012

(54) ANTIMICROBIAL COMPOSITIONS AND Publication Classification METHODS OF USE THEREFORE (51) Int. Cl. (75) Inventors: Robert Moir, Medfield, MA (US); st 7, CR Rudolph E. Tanzi, Hull, MA (US) (2006.01) AOIP I/00 (2006.01) (73) Assignee: The General Hospital A6IP3L/2 (2006.01) Corporation, Boston, MA (US) AOIP3/00 (2006.01) AOIN 37/18 (2006.01) (21) Appl. No.: 13/256,171 A6IP3 L/10 (2006.01) (22) PCT Filed: Mar. 12, 2010 (52) U.S. Cl...... 514/2.6; 514/2.3: 514/24: 514/3.3: (86). PCT No.: PCT/US 10/27186 514/3.7: 514/3.4: 514/2.8: 514/2.7 S371 (c)(1), (2), (4) Date: Nov. 18, 2011 (57) ABSTRACT O O The invention features antimicrobial compositions compris Related U.S. Application Data ing B-amyloid peptides, oligomers, and analogs thereof, and (60) Provisional application No. 61/159,671, filed on Mar. methods of using them for the prevention or treatment of an 12, 2009. infection. Patent Application Publication Mar. 8, 2012 Sheet 1 of 5 US 2012/0058935 A1

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ANTIMICROBAL COMPOSITIONS AND 0011. In various embodiments of any of the aspects delin METHODS OF USE THEREFORE eated herein, the B-amyloid peptide, oligomer, derivative or analog thereof, is formulated as a skin cleanser. In various CROSS-REFERENCE TO RELATED embodiments of any of the aspects delineated herein, the skin APPLICATION is cleansed in preparation for Surgery. 0012. In various embodiments of any of the aspects delin 0001. This application claims the benefit of the following eated herein, the microbe is a bacteria, virus, or fungus. In U.S. Provisional Application No. 61/159,671, filed Mar. 12, various embodiments of any of the aspects delineated herein, 2009, the entire contents of which are incorporated herein by microbe is one or more of Candida albicans, Escherichia reference. coli, Staphylococcus epidermidis, Streptococcus pneumo niae, Staphylococcus aureus, Listeria monocytogenes, BACKGROUND OF THE INVENTION Enterococcus faecalis, Streptococcus agalactiae Pseudomo 0002 The amyloid?-protein (AB) is believed to be the key nas aeruginosa, Streptococcus pyogenes, Streptococcus mediator of Alzheimer's disease (AD) pathology. A? is most mitis, and Streptococcus salivariu. often characterized as an incidental catabolic byproduct that 0013. In various embodiments of any of the aspects delin lacks a normal physiological role. However, AB has been eated herein, the method cleans, sanitizes, or disinfects the shown to be a specific ligand for a number of different recep Surface. In various embodiments, the Surface is a wall, floor, tors and other molecules, transported by complex trafficking ceiling, counter, machine, or other Surface or equipment pathways, modulated in response to a variety of environmen present in an industrial, commercial, or clinical setting. In tal stressors, and able to induce pro-inflammatory activities. various embodiments, the clinical setting is a hospital, medi cal office, clinic, health center, or laboratory. In other embodi ments, the Surface contacted is present in a dwelling, School, SUMMARY OF THE INVENTION office building, plane, train, automobine, restaurant, cafete 0003. As described below, the present invention features ria, or daycare center. antimicrobial compositions and methods of using them for 0014. The invention provides antimicrobial compositions the prevention or treatment of an infection. and methods of using them for the prevention or treatment of 0004. In one aspect, the invention provides a method for an infection. Compositions and articles defined by the inven treating or preventing a microbial infection in a Subject in tion were isolated or otherwise manufactured in connection need of treatment or prevention involving contacting the Sub with the examples provided below. Other features and advan ject with an effective amount of one or more of a B-amyloid tages of the invention will be apparent from the detailed peptide, oligomer, derivative or analog thereof, thereby pre description, and from the claims. venting or inhibiting a microbial infection in the Subject. 0005. In another aspect, the invention provides a method DEFINITIONS for treating or preventing a skin infection in a Subject involv 0015. By “B-amyloid peptide' is meant a 4.2-kD polypep ing administering to the Subject an effective amount of one or tide having a 37-43-amino acid sequence present in amyloid more of a B-amyloid peptide, oligomer, derivative or analog plaques and soluble aggregates, which accumulate in the thereof, in an amount sufficient to treat the subject. brains of patients with Alzheimer's disease. Exemplary AB 0006. In yet another aspect, the invention provides a peptides include peptides having the following amino acid method for preventing or inhibiting microbial growth on a Surface, involving contacting the Surface with an effective Sequences: amount of one or more of a B-amyloid peptide, oligomer, derivative or analog thereof, thereby preventing or inhibiting DAEFRHDSGYEVHHOKLVFFAEDWGSNKGAIIGLMVGGVWIAT microbial growth on the Surface. 0007. In still another aspect, the invention provides a (A343 ); method of sanitizing or disinfecting a body part involving contacting the body part with an effective amount of a B-amy DAEFRHDSGYEVHHOKLVFFAEDVGSNKGAIIGLMVGGVVIA (AB42) ; loid peptide, oligomer, derivative or analog thereof, thereby DAEFRHDSGYEVHHOKLVFFAEDVGSNKGAIIGLMVGGVV (AB40); sanitizing or disinfecting the body part. 0008. In one aspect, the invention provides an antimicro DAEFRHDSGYEVHHOKLVFFAEDVGSNKGAIIGLMVG (AB37). bial composition containing an effective amount of a B-amy 0016. By “anti-microbial composition' is meant any com loid peptide, oligomer, derivative or analog thereof, thereof in position that prevents, inhibits, slows, or reduces the growth, a carrier or diluent. proliferation, or Survival of a microbe (e.g., bacteria, fungus 0009. In another aspect, the invention provides a pharma (e.g., mold), protozoa, virus). ceutical pack containing a B-amyloid peptide, oligomer, 0017. By "agent' is meant any small molecule chemical derivative or analog thereof, in an individual dosage amount. compound, antibody, nucleic acid molecule, or polypeptide, 0010. In various embodiments of any of the aspects delin or fragments thereof. eated herein, the effective amount comprises 1% to 90% of a 0018. By “ameliorate” is meant decrease, suppress, B-amyloid peptide, oligomer, derivative or analog thereof, or attenuate, diminish, arrest, or stabilize the development or combinations thereof. In various embodiments of any of the progression of a disease. aspects delineated herein, the B-amyloid peptide, oligomer, 0019. By “alteration' is meant a change (increase or derivative or analog thereof, is formulated as a water soluble decrease) in the expression levels or activity of a gene or or water insoluble solid, liquid, emulsion, slurry, or powder. polypeptide as detected by standard art known methods such In various embodiments, the composition is topically admin as those described herein. As used herein, an alteration istered. includes a 10% change in expression levels, preferably a 25% US 2012/0058935 A1 Mar. 8, 2012 change, more preferably a 40% change, and most preferably 0030. By “inhibiting microbial growth' is meant prevent a 50% or greater change in expression levels.” ing, slowing, or otherwise reducing the proliferation of a 0020. By “analog is meant a molecule that is not identi microbe. Such inhibition is by at least about 5%, 10%, 25%, cal, but has analogous functional or structural features. For 50%, 75%, or 100%. example, a polypeptide analog retains the biological activity 0031. By “home antimicrobial is meant an antimicrobial of a corresponding naturally-occurring polypeptide, while composition used to clean, sanitize, or disinfect a surface with having certain biochemical modifications that enhance the a home. In various embodiments, home antimicrobials are analog's function relative to a naturally occurring polypep used in a kitchens, bathrooms, and other rooms within a tide. Such biochemical modifications could increase the ana dwelling to prevent or inhibit microbial growth on walls, log's protease resistance, membrane permeability, or half floors, carpets, ceilings, counters, appliances, food prepara life, without altering, for example, ligand binding. An analog tion or storage vessels, or other implements used in food may include an unnatural amino acid. preparation. 0021. By “body part is meant any tissue or organ of a mammalian or avian organism. Exemplary body parts 0032. By "isolated polynucleotide' is meant a nucleic acid include, but are not limited to, skin, hair, fur, epidermis, (e.g., a DNA) that is free of the genes which, in the naturally feathers, wool, hide, the oral cavity, including but not limited occurring genome of the organism from which the nucleic to, the lips, tongue, and teeth. acid molecule of the invention is derived, flank the gene. The 0022. By “dwelling' is meant any human residence. term therefore includes, for example, a recombinant DNA 0023. In this disclosure, "comprises.” “comprising.” “con that is incorporated into a vector; into an autonomously rep taining and “having and the like can have the meaning licating plasmid or virus; or into the genomic DNA of a ascribed to them in U.S. Patent law and can mean “includes.” prokaryote or eukaryote; or that exists as a separate molecule “including,” and the like: “consisting essentially of or “con (for example, a cDNA or a genomic or cDNA fragment pro sists essentially” likewise has the meaning ascribed in U.S. duced by PCR or restriction endonuclease digestion) inde Patent law and the term is open-ended, allowing for the pres pendent of other sequences. In addition, the term includes an ence of more than that which is recited so long as basic or RNA molecule that is transcribed from a DNA molecule, as novel characteristics of that which is recited is not changed by well as a recombinant DNA that is part of a hybrid gene the presence of more than that which is recited, but excludes encoding additional polypeptide sequence. prior art embodiments. 0033. By an "isolated polypeptide' is meant a polypeptide 0024 “Detect” refers to identifying the presence, absence of the invention that has been separated from components that or amount of the analyte to be detected. naturally accompany it. Typically, the polypeptide is isolated 0025 By “disease' is meant any condition or disorder that when it is at least 60%, by weight, free from the proteins and damages or interferes with the normal function of a cell, naturally-occurring organic molecules with which it is natu tissue, or organ. Examples of diseases include bacterial inva rally associated. Preferably, the preparation is at least 75%, sion or colonization of a host cell. more preferably at least 90%, and most preferably at least 0026. By “effective amount” is meant an amount of a 99%, by weight, a polypeptide of the invention. An isolated compound that prevents, inhibits, slows, stabilizes, or reduces polypeptide of the invention may be obtained, for example, by the growth of a microbe. An effective amount of the com extraction from a natural Source, by expression of a recom pound described above may range from about 0.1 ppm to binant nucleic acid encoding Such a polypeptide; or by chemi about 1000 ppm. Effective amounts may vary depending on cally synthesizing the protein. Purity can be measured by any the microbe to be inhibited, the application method, as well as appropriate method, for example, column chromatography, the possibility of co-usage with other agents. polyacrylamide gel electrophoresis, or by HPLC analysis. 0027. The invention provides a number of targets that are 0034. By “marker' is meant any protein or polynucleotide useful for the development of highly specific drugs to treat or having an alteration in expression level or activity that is a disorder characterized by the methods delineated herein. In associated with a disease or disorder. addition, the methods of the invention provide a facile means 0035. As used herein, “obtaining as in “obtaining an to identify therapies that are safe for use in subjects. In addi agent includes synthesizing, purchasing, or otherwise tion, the methods of the invention provide a route for analyz acquiring the agent. ing virtually any number of compounds for effects on a dis 0036 By “microbe' is meant a bacterium, fungus, proto ease described herein with high-volume throughput, high Zoa, virus, or other microscopic organism. Microbes include sensitivity, and low complexity. airborne pathogens. 0028 By “fragment' is meant a portion of a polypeptide or 0037. By “pathogen' is meant any bacteria, viruses, fungi, nucleic acid molecule. This portion contains, preferably, at or protozoans capable of interfering with the normal function least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of of a cell. the entire length of the reference nucleic acid molecule or 0038 Exemplary bacterial pathogens include, but are not polypeptide. A fragment may contain 10, 20, 30, 40, 50, 60, limited to, Aerobacter, Aeromonas, Acinetobacter, Agrobac 70, 80,90, or 100, 200, 300, 400, 500, 600, 700, 800, 900, or terium, Bacillus, Bacteroides, Bartonella, Bordtella, Bru 1000 nucleotides or amino acids. cella, Burkholderia, Calymmatobacterium, Campylobacter, 0029. By “industrial antimicrobial' is meant an antimicro Citrobacter, Clostridium, Cornyebacterium, Enterobacter, bial composition used to clean, sanitize, or disinfect a Surface Escherichia, Francisella, Haemophilus, Hafinia, Helico in any industrial setting. In various embodiments, industrial bacter; Klebsiella, Legionella, Listeria, Morganella, antimicrobials are used on walls, floors, ceilings, counters, Moraxella, Proteus, Providencia, Pseudomonas, Salmonella, and machinery present in factories, food processing facilities, Serratia, Shigella, Staphylococcus, Streptococcus, Tre and hospitals. ponema, Xanthomonas, Vibrio, and Yersinia. US 2012/0058935 A1 Mar. 8, 2012

0039. By “protective immune response' is meant an peratures of at least about 30°C., more preferably of at least immune response Sufficient to ameliorate a pathogen infec about 37° C., and most preferably of at least about 42° C. tion in a mammal. Varying additional parameters, such as hybridization time, 0040. As used herein, the terms “prevent.”99 “preventing.”&g the concentration of detergent, e.g., sodium dodecyl Sulfate “prevention.” “prophylactic treatment” and the like refer to (SDS), and the inclusion or exclusion of carrier DNA, are well reducing the probability of developing a disorder or condition known to those skilled in the art. Various levels of stringency in a subject, who does not have, but is at risk of or susceptible are accomplished by combining these various conditions as to developing a disorder or condition. needed. In a preferred: embodiment, hybridization will occur 0041) “Primer set’ means a set of oligonucleotides that at 30°C. in 750 mM. NaCl, 75 mM trisodium citrate, and 1% may be used, for example, for PCR. A primer set would SDS. In a more preferred embodiment, hybridization will consist of at least 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 30, 40, 50, occurat 37°C. in 500 mMNaCl, 50mMtrisodium citrate, 1% 60, 80, 100, 200, 250, 300, 400, 500, 600, or more primers. SDS, 35% formamide, and 100 ug/ml denatured salmon 0042. By “reduces” is meant a negative alteration of at sperm DNA (ssDNA). In a most preferred embodiment, least 10%, 25%, 50%, 75%, or 100%. hybridization will occur at 42°C. in 250 mM. NaCl, 25 mM 0043. By “reference' is meant a standard or control con trisodium citrate, 1% SDS, 50% formamide, and 200 g/ml dition. ssDNA. Useful variations on these conditions will be readily 0044. A “reference sequence' is a defined sequence used apparent to those skilled in the art. as a basis for sequence comparison. A reference sequence 0047 For most applications, washing steps that follow may be a Subset of or the entirety of a specified sequence; for hybridization will also vary in Stringency. Wash stringency example, a segment of a full-length cDNA or gene sequence, conditions can be defined by Salt concentration and by tem or the complete cDNA or gene sequence. For polypeptides, perature. As above, wash Stringency can be increased by the length of the reference polypeptide sequence will gener decreasing salt concentration or by increasing temperature. ally be at least about 16 amino acids, preferably at least about For example, stringent salt concentration for the wash steps 20 amino acids, more preferably at least about 25 amino will preferably be less than about 30 mM. NaCl and 3 mM acids, and even more preferably about 35 amino acids, about trisodium citrate, and most preferably less than about 15 mM 50 amino acids, or about 100 amino acids. For nucleic acids, NaCl and 1.5 mM trisodium citrate. Stringent temperature the length of the reference nucleic acid sequence will gener conditions for the wash steps will ordinarily include a tem ally be at least about 50 nucleotides, preferably at least about perature of at least about 25°C., more preferably of at least 60 nucleotides, more preferably at least about 75 nucleotides, about 42°C., and even more preferably of at least about 68°C. and even more preferably about 100 nucleotides or about 300 In a preferred embodiment, wash steps will occur at 25°C. in nucleotides or any integer thereabout or therebetween. 30 mM. NaCl, 3 mM trisodium citrate, and 0.1% SDS. In a 0.045 Nucleic acid molecules useful in the methods of the more preferred embodiment, wash steps will occur at 42 C in invention include any nucleic acid molecule that encodes a 15 mM NaCl, 1.5 mM trisodium citrate, and 0.1% SDS. In a polypeptide of the invention or a fragment thereof. Such more preferred embodiment, wash steps will occur at 68°C. nucleic acid molecules need not be 100% identical with an in 15 mM NaCl, 1.5 mM trisodium citrate, and 0.1% SDS. endogenous nucleic acid sequence, but will typically exhibit Additional variations on these conditions will be readily substantial identity. Polynucleotides having “substantial apparent to those skilled in the art. Hybridization techniques identity” to an endogenous sequence are typically capable of are well knownto those skilled in the art and are described, for hybridizing with at least one strand of a double-stranded example, in Benton and Davis (Science 196:180, 1977); nucleic acid molecule. Nucleic acid molecules useful in the Grunstein and Hogness (Proc. Natl. Acad. Sci., USA methods of the invention include any nucleic acid molecule 72:3961, 1975); Ausubeletal. (Current Protocols in Molecu that encodes a polypeptide of the invention or a fragment lar Biology, Wiley Interscience, New York, 2001); Berger and thereof. Such nucleic acid molecules need not be 100% iden Kimmel (Guide to Molecular Cloning Techniques, 1987, tical with an endogenous nucleic acid sequence, but will Academic Press, New York); and Sambrook et al., Molecular typically exhibit substantial identity. Polynucleotides having Cloning: A Laboratory Manual, Cold Spring Harbor Labora “Substantial identity” to an endogenous sequence are typi tory Press, New York. cally capable of hybridizing with at least one strand of a 0048. By “substantially identical' is meant a polypeptide double-stranded nucleic acid molecule. By “hybridize' is or nucleic acid molecule exhibiting at least 50% identity to a meant pair to form a double-stranded molecule between reference amino acid sequence (for example, any one of the complementary polynucleotide sequences (e.g., a gene amino acid sequences described herein) or nucleic acid described herein), or portions thereof, under various condi sequence (for example, any one of the nucleic acid sequences tions of stringency. (See, e.g., Wahl, G. M. and S. L. Berger described herein). Preferably, such a sequence is at least 60%, (1987) Methods Enzymol. 152:399; Kimmel, A. R. (1987) more preferably 80% or 85%, and more preferably 90%.95% Methods Enzymol. 152:507). or even 99% identical at the amino acid level or nucleic acid 0046 For example, stringent salt concentration will ordi to the sequence used for comparison. narily be less than about 750 mMNaCl and 75 mM trisodium 0049 Sequence identity is typically measured using citrate, preferably less than about 500 mM. NaCl and 50 mM sequence analysis Software (for example, Sequence Analysis trisodium citrate, and more preferably less than about 250 Software Package of the Genetics Computer Group, Univer mM NaCl and 25 mM trisodium citrate. Low stringency sity of Wisconsin Biotechnology Center, 1710 University hybridization can be obtained in the absence of organic Sol Avenue, Madison, Wis. 53705, BLAST, BESTFIT, GAP, or vent, e.g., formamide, while high Stringency hybridization PILEUP/PRETTYBOX programs). Such software matches can be obtained in the presence of at least about 35% forma identical or similar sequences by assigning degrees of homol mide, and more preferably at least about 50% formamide. ogy to various Substitutions, deletions, and/or other modifi Stringent temperature conditions will ordinarily include tem cations. Conservative substitutions typically include Substi US 2012/0058935 A1 Mar. 8, 2012

tutions within the following groups: glycine, alanine; Valine, washes, the bacteria were fixed onto glass slides and immu isoleucine, leucine; aspartic acid, glutamic acid, asparagine, nostained with the HRP conjugated anti-A? antibody (mAb glutamine; serine, threonine; lysine, arginine; and phenylala 6E10-HRP). nine, tyrosine. In an exemplary approach to determining the degree of identity, a BLAST program may be used, with a 0059 FIGS. 3A and 3B are graphs showing that AD brain probability score betweene and e' indicating a closely homogenates have increased antimicrobial activity against C. related sequence. albicans. AD and non-AD brain samples were tested for 0050. By “subject' is meant a mammal, including, but not Af-mediated inhibition of C. albicans. Samples of temporal limited to, a human or non-human mammal. Such as a bovine, lobe (Temp. L.) and cerebellum (Cereb.) from AD (n=32) and equine, canine, Ovine, or feline. age-matched control Subjects (n=13) were homogenized in 0051 Ranges provided herein are understood to be short culture broth. In FIG. 3A, homogenates were inoculated with hand for all of the values within the range. For example, a log-phase C. albicans and microbial growth determined by range of 1 to 50 is understood to include any number, com alamar blue viability assay. Data is shown as percentage of bination of numbers, or Sub-range from the group consisting signal for C. albicans alone (average of four replicates)ts.e. 1,2,3,4,5,6,7,8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, m. In FIG. 3B, homogenates were assayed for AB40 and 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32,33, 34,35, 36, 37, Af342 by commercially available ELISA. The graph in FIG. 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50. 3B shows AB signal (sum of AB40 and AB42) against C. 0052. As used herein, the terms “treat,” treating.” “treat albicans growth for temporal lobe homogenates from com ment, and the like refer to reducing or ameliorating a disor bined AD and non-demented cohorts (n=42). Probability der and/or symptoms associated therewith. It will be appre analysis used unpaired two-tailed t-tests (p). Correlation was ciated that, although not precluded, treating a disorder or determined by calculating the Pearson r correlation coeffi condition does not require that the disorder, condition or cient (r). symptoms associated therewith be completely eliminated. 0060 FIGS. 4A and 4B depict results showing that immu 0053. Unless specifically stated or obvious from context, nodepletion of A? from AD brain homogenates attenuates C. as used herein, the term 'or' is understood to be inclusive. albicans inhibition. Homogenates of temporal lobe (Temp. Unless specifically stated or obvious from context, as used L.) and cerebellum (Cereb.) were prepared from AD (n=32) herein, the terms “a”, “an', and “the are understood to be or non-demented (n=13) subjects. AD (AD) or non-demented singular or plural. (non-AD) homogenates were pooled and then incubated with 0054) Unless specifically stated or obvious from context, Magno-beads pre-loaded with rabbit IgG (IgG) or a poly as used herein, the term “about is understood as within a range of normal tolerance in the art, for example within 2 conal rabbit anti-A? antibody (C.-AB). Following bead standard deviations of the mean. About can be understood as removal samples were analyzed for AB signal by Western blot within 10%, 9%, 8%, 7%, 6%. 5%, 4%, 3%, 2%, 1%, 0.5%, and assayed for C. albicans growth by alamar blue viability 0.1%, 0.05%, or 0.01% of the stated value. Unless otherwise assay. FIG. 4A shows C. albicans growth in treated homoge clear from context, all numerical values provided herein are nates as a percentage of signal in culture broth alone. Immu modified by the term about. nodepletion of AD temporal lobe homogenates with C.-AB 0055. The recitation of a listing of chemical groups in any restored microbial growth to levels equivalent to non-de definition of a variable herein includes definitions of that mented control samples. Graph shows average of five variable as any single group or combination of listed groups. replicatests.e.m. Statistical probability analysis (p) of data The recitation of an embodiment for a variable or aspect used unpaired two-tailed t-test. In FIG. 4B, untreated and herein includes that embodiment as any single embodiment immunodepleted homogenates (1:16 dilution) were Western or in combination with any other embodiments or portions blotted and probed with the AB-specific mAb 4G8 antibody. thereof. Analysis confirmed AB signal was reduced in temporal lobe 0056. Any compositions or methods provided herein can homogenate incubated with anti-amyloid B-peptide antibody be combined with one or more of any of the other composi (Lane 1) compared to sample incubate alone (Lane 2) or with tions and methods provided herein. rabbit IgG (Lane 3). A? in dilutions of cerebellum homoge nate is below the level of detection for our experimental BRIEF DESCRIPTION OF THE DRAWINGS conditions (Lanes 4-6). Blots included synthetic Af42 (AB42) standard (Lane 7). 0057 FIG. 1 is a graph showing that growth of E. faecalis is inhibited by AB42. E. faecalis were cultured alone (circle) 0061 FIG. 5 is a graph showing that AB-mediated inhibi with 25 ug/ml of AB42 (triangle) or LL-37 (diamond). Bac tion of C. albicans in AD brain homogenates is dose depen terial growth with time was monitored by inoculation of agar dant. AD temporal lobe (Temp. L.) or cerebellum (Cereb.) with diluted incubants and counting CFU. Representative were homogenized in phosphate buffer. Temporal lobe data from six experiments is shown as mean signal of four (n=30.) or cerebellum (n=32) homogenates were pooled and replicatests.e.m. At the time points indicated in the graph, 1:16, 1:32, and 1:64 serial dilutions prepared in culture broth. incubants were monitored for AB42 and LL-37 by Western Homogenate dilutions were incubated with mouse IgG (IgG) blot with mAb 6E10 or anti-LL-37. The figure shows repre or anti-AB mAb 6E10 (6E10) antibody immobilized on sentative signal for AB42 (odd lanes) or LL-37 (even lanes) MagnaBind beads. Following pelleting of the beads incu incubants from six replicate experiments. bants were inoculated with mid-logarithmic phase C. albi 0058 FIGS. 2A and 2B are images showing that E. faeca cans in 96-well plates. Microbial growth was determined by lis pre-incubated with AB42 are mab 6E10 immunoreactive. alamar blue cell viability assay. Graphs shows percentage Bacteria were incubated (1 hr at 37° C.) with (FIG. 2A) or signal of C. albicans alone (average of five replicates)ts.e.m. without (FIG. 2B) A42 (25 ug/ml). Following repeated Consistent with AB-mediated antimicrobial activity, C. albi US 2012/0058935 A1 Mar. 8, 2012

cans growth is highest for samples with low AB levels and detail below, LL-37 exhibits striking similarities to AB, increases with homogenate dilution. including a propensity to form cytotoxic soluble oligomers 15,16,17.18 and insoluble fibrils that demonstrate congo DETAILED DESCRIPTION OF THE INVENTION philia and birefringence 19, two classical histochemical properties of tinctorial amyloid. While the microbiocidal 0062. The invention features antimicrobial compositions activity of LL-37 has been well characterized 20, the activ and related prophylactic and therapeutic methods. ity of AB against microbial organisms has not been tested. 0063. The invention is based, at least in part, on the dis 0066. As reported herein, AB is active against at least eight covery that amyloid 3-protein (AB) is a hitherto unrecognized common and clinically relevant microorganisms. The in vitro antimicrobial peptide (AMPs) that normally functions in the antimicrobial activity of AB matched, and in some cases, innate immune system. This finding stands in Stark contrast to exceeded, that of LL-37, an archetypical human AMP. Fur current models of AB-mediated pathology and has important thermore, anti-A?s immunoreactive material in AD whole implications for ongoing and future Alzheimer disease treat brain homogenates is active against Candida albicans, the ment strategies. As reported in more detail below, in vitro pathogen identified herein as most sensitive to synthetic A13. assays were used to compare the antimicrobial activities of Most strikingly, temporal lobe samples from AD brain con A? and LL-37, an archetypical human AMP. AB was found to tained significantly higher antimicrobial activity than mate have antimicrobial activity against eight common and clini rial from the same brain area of aged-matched, non-ADSub cally relevant microorganisms with a potency equivalent to, jects. Consistent with an Af-mediated action, cerebellum and in Some cases greater than, LL-37. Furthermore, AD samples with low B-amyloid loads from the same set of whole brain homogenates have significantly higher antimi affected and unaffected subjects were not significantly differ crobial activity thanaged matched non-AD samples and AMP ent with regards to antimicrobial activity. These findings action correlates with tissue AB levels. Consistent with Af show A3 possesses antimicrobial activity and may function in mediated activity, the increased antimicrobial action was vivo as an AMP and, thus, play a role as an effector molecule ablated by immunodepletion of AD brain homogenates with of innate immunity. anti-AB antibodies. 0067. Accordingly, the invention features antimicrobial compositions containing B amyloid peptides and methods Amyloid f-Peptide employing such compositions for the treatment and preven 0064. The past 25 years has witnessed the accrual of a tion of pathogen infections. large body of data concerning the physiochemistry and bio logical activities of the amyloid B-peptide (AB), the main Amyloid Beta Peptide and Analogs component of 3-amyloid deposits in the brains of Alzhe 0068 Also included in the invention are beta amyloid imer's disease (AD) patients. Af, which is generated in the peptides, oligomers, and fragments thereof that are modified brain and peripheral tissues, is widely believed to be an inci in ways that enhance or do not inhibit their antimicrobial dental catabolic byproduct of the amyloid B protein precursor activity. In one embodiment, the invention provides methods (APP) with no normal physiological function. A? has been for optimizing an Bamyloid peptide amino acid sequence or shown to be a ligand for a number of different receptors and nucleic acid sequence by producing an alteration. Such other molecules 2.3.4. transported by complex trafficking changes may include certain mutations, deletions, insertions, pathways between tissues and across the blood brain barrier or post-translational modifications. The invention further 1.5, modulated in response to a variety of environmental includes analogs of any naturally-occurring polypeptide of stressors, and able to induce pro-inflammatory activities the invention. Analogs can differ from the naturally-occurring 6.7. Nevertheless, the normal physiological role of Af polypeptide of the invention by amino acid sequence differ remains unknown. Many of the physiochemical and biologi ences, by post-translational modifications, or by both. Ana cal properties previously reported for AB are similar to those logs of the invention will generally exhibit at least 85%, more of a group of biomolecules collectively known as “antimicro preferably 90%, and most preferably 95% or even 99% iden bial peptides’ (AMPs) which function in the innate immune tity with all or part of a naturally-occurring amino, acid system. AMPs (also called “host defense peptides’) are sequence of the invention. The length of sequence compari potent, broad-spectrum antibiotics that target Gram-negative son is at least 10, 13, 15 amino acid residues, preferably at and Gram-positive bacteria, mycobacteria, enveloped least 25 amino acid residues, and more preferably more than viruses, fungi, protozoans and in Some cases, transformed or 35 amino acid residues. Again, in an exemplary approach to cancerous host cells. AMPs are also potent immunomodula determining the degree of identity, a BLAST program may be tors that mediate cytokine release and adaptive immune used, with a probability score between eande' indicat responses (see review by Zaiou, 20078). ing a closely related sequence. Modifications include in vivo 0065. The three main families of mammalian AMPs are and in vitro chemical derivatization of polypeptides, e.g., the defensins, the histatins, and the cathelicidins. Only one acetylation, carboxylation, phosphorylation, or glycosyla member of the cathelicidin family has been identified in tion; Such modifications may occur during polypeptide Syn humans, the LL-37 peptide 9. The pleiotropic LL-37 pep thesis or processing or following treatment with isolated tide is a widely expressed archetypal AMP 10. The rodent modifying enzymes. Analogs can also differ from the natu LL-37 homologue (CRAMP) has been shown to play a cen rally-occurring polypeptides of the invention by alterations in tral role in combating bacterial infections in a range of tissues, primary sequence. These include genetic variants, both natu including the CNS 11. Patients that express low levels of ral and induced (for example, resulting from random LL-37 are at increased risk for serious infections 12. Con mutagenesis by irradiation or exposure to ethanemethylsul versely, high levels of LL-37 are associated with the pathol fate or by site-specific mutagenesis as described in Sam ogy of several presumably non-infectious diseases 13. brook, Fritsch and Maniatis, Molecular Cloning: A Labora including plaques in atherosclerosis 14. As reported in more tory Manual (2d ed.). CSH Press, 1989, or Ausubel et al., US 2012/0058935 A1 Mar. 8, 2012

Supra). Also included are cyclized peptides, molecules, and cida, Bacteroides sp., Fusobacterium nucleatum, analogs which contain residues other than L-amino acids, Streptobacillus moniliformis, Treponema pallidium, Tre e.g., D-amino acids or non-naturally occurring or synthetic ponema pertenue, Leptospira, Rickettsia ssp. Yersinia pestis amino acids, e.g., B or Yamino acids. and Actinomyces israeli. 0069. In addition to full-length polypeptides, the invention 0072. In other embodiments, the invention provides for the also includes fragments of any one of the polypeptides of the treatment or prevention of a protist infection. Such organisms invention. As used herein, the term “a fragment’ means at include Plasmodium spp. Such as Plasmodium falciparum, least 5, 10, 13, or 15. In other embodiments a fragment is at Plasmodium malariae, Plasmodium ovale, and Plasmodium least 20 contiguous amino acids, at least 30 contiguous amino vivax and Toxoplasma gondii. Blood-borne and/or tissues acids, or at least 50 contiguous amino acids, and in other parasites include Plasmodium spp., Babesia microti, Babesia embodiments at least 60 to 80 or more contiguous amino divergens, Leishmania tropica, Leishmania spp., Leishmania acids. Fragments of the invention can be generated by meth braziliensis, Leishmania donovani, Trypanosoma gambiense ods known to those skilled in the art or may result from and Trypanosoma rhodesiense (African sleeping sickness), normal protein processing (e.g., removal of amino acids from Trypanosoma Cruzi (Chagas disease), and Toxoplasma gon the nascent polypeptide that are not required for biological dii. activity or removal of amino acids by alternative mRNA 0073. In still other embodiments, the invention provides splicing or alternative protein processing events). for the treatment or prevention of an infection with a patho 0070. Non-protein B amyloid peptide analogs having a genic fungus, including, without limitation, Alternaria, chemical structure designed to mimic B amyloid peptides Aspergillus, Basidiobolus, Bipolaris, Blastoschizomyces, functional activity can be administered according to methods Candida, Candida albicans, Candida krusei, Candida gla of the invention. 3 amyloid peptide analogs may exceed the brata (formerly called Torulopsis glabrata), Candida parap antimicrobial activity of native beta amyloid peptides. Meth silosis, Candida tropicalis, Candida pseudotropicalis, Can ods of analog design are well known in the art, and synthesis dida guilliermondii, Candida dubliniensis, and Candida of analogs can be carried out according to Such methods by lusitaniae, Coccidioides, Cladophialophora, Cryptococcus, modifying the chemical structures such that the resultant Cunninghamella, Curvularia, Exophiala, Fonsecaea, Histo analogs exhibit the antimicrobial activity of a native beta plasma, Madurella, Malassezia, Plastomyces, Rhodotorula, amyloid peptide. These chemical modifications include, but Scedosporium, Scopulariopsis, Sporobolomyces, Tinea, and are not limited to, Substituting alternative R groups and vary TrichospOron. ing the degree of Saturation at specific carbon atoms of the 0074 Fungi, including, but not limited to Candida, cause native Bamyloid peptide molecule. Preferably, the Bamyloid invasive diseases in hosts with altered immunity, such as peptide analogs are relatively resistant to in vivo degradation, patients with HIV infection, organ or bone marrow trans resulting in a more prolonged therapeutic effect upon admin plants, or neutropenia following cancer immunotherapy. istration. Assays for measuring functional activity include, There are approximately 200 species of the genus Candida, but are not limited to, those described below and in the but nine cause the great majority of human infections. They Examples. are C. albicans, C. krusei, C. glabrata (formerly called Toru lopsis glabrata), C. parapsilosis, C. tropicalis, C. pseudotro Anti-Microbial Activity picalis, C. guilliermondii, C. dubliniensis, and C. lusitaniae. 0071. The invention provides compositions comprising They cause infections of the mucous membranes, for B-amyloid peptides, oligomers, and analogs thereof for the example, thrush, esophagitis, and vagititis; skin, for example, treatment or prevention of infections with any of a number of intertrigo, balanitis, and generalized candidiasis; blood pathogens, including but not limited to bacteria, viruses, pro stream infections, for example, candidemia; and deep organ tists, and fungi. In one embodiment, the invention provides infections, for example, hepatosplenic candidiasis, urinary for the treatment or prevention of bacterial infections, includ tract candidiasis, arthritis, endocarditis, and endophthamitis. ing infections with gram negative and gram positive bacteria, 0075. In still other embodiments, the methods of the which serve as antigens in Vertebrate animals. Such gram invention can be used to treat or prevent a viral infection. positive bacteria include, but are not limited to, Pasteurella Accordingly, the invention provides a composition compris species, Staphylococci species, and Streptococcus species. ing a B-amyloid peptides, oligomers, and analogs thereofthat Gram negative bacteria include, but are not limited to, inhibits the growth of a viral pathogen. Exemplary viral Escherichia coli, Pseudomonas species, and Salmonella spe pathogens include but are not limited to: Retroviridae (e.g. cies. Specific examples of infectious bacteria include but are human immunodeficiency viruses, such as HIV-1 (also not limited to, Helicobacter pyloris, Burkholderia sps, Borel referred to as HDTV-III, LAVE or HTLV-III/LAV, or HIV-III: lia burgdorferi, Legionella pneumophilia, Mycobacteria sps and other isolates, such as HIV-LP: Picornaviridae (e.g. polio (e.g. M. tuberculosis, M. avium, M. intracellulare, M. kansai, viruses, hepatitis. A virus; enteroviruses, human Coxsackie M. gordonae), Staphylococcus aureus, Neisseria gonor viruses, rhinoviruses, echoviruses); Calciviridae (e.g. strains rhoeae, Neisseria meningitidis, Listeria monocytogenes, that cause gastroenteritis); Togaviridae (e.g. equine encepha Streptococcus pyogenes (Group A Streptococcus), Strepto litis viruses, rubella viruses); Flaviridae (e.g. dengue viruses, coccus agalactiae (Group B Streptococcus), Streptococcus encephalitis viruses, yellow fever viruses); Coronoviridae (viridans group), Streptococcus faecalis, Streptococcus (e.g. coronaviruses); Rhabdoviridae (e.g. vesicular stomatitis bovis, Streptococcus (anaerobic sps.). Streptococcus pneu viruses, rabies viruses); Filoviridae (e.g. ebola viruses); moniae, pathogenic Campylobacter sp., Enterococcus sp., Paramyxoviridae (e.g. parainfluenza viruses, mumps virus, Haemophilus influenzae, Bacillus antracis, corynebacterium measles virus, respiratory syncytial virus); Orthomyxoviri diphtheriae, corynebacterium sp., Erysipelothrix rhusio dae (e.g. influenza viruses); Bungaviridae (e.g. Hantaan pathiae, Clostridium perfiringens, Clostridium tetani, Entero viruses, bunga viruses, phleboviruses and Nairo viruses); bacter aerogenes, Klebsiella pneumoniae, Pasturella multo Arena viridae (hemorrhagic fever viruses); Reoviridae (e.g. US 2012/0058935 A1 Mar. 8, 2012

reoviruses, orbiviurses and rotaviruses); Birnaviridae, Hep nation of the first level, and the two levels are compared to admaviridae (Hepatitis B virus); Parvovirida (parvoviruses): monitor the course of disease or the efficacy of the therapy. In Papovaviridae (papilloma viruses, polyoma viruses); Aden certain preferred embodiments, a pre-treatment level of Oviridae (most adenoviruses); Herpesviridae (herpes simplex Marker in the Subject is determined prior to beginning treat virus (HSV) 1 and 2, varicella Zoster virus, cytomegalovirus ment according to this invention; this pre-treatment level of (CMV), herpes virus; Poxviridae (variola viruses, vaccinia Marker can then be compared to the level of Marker in the viruses, pox viruses); and Iridoviridae (e.g. African Swine subject after the treatment commences, to determine the effi fever virus); and unclassified viruses (e.g. the agent of delta cacy of the treatment. hepatitis (thought to be a defective satellite of hepatitis B virus), the agents of non-A, non-B hepatitis (class 1 inter Skin Infections nally transmitted; class 2 parenterally transmitted (i.e. Hepa 0080 Compositions of the invention are particularly use titis C); Norwalk and related viruses, and astroviruses). ful as antimicrobials to prevent, slow, or otherwise inhibit the 0076. The present invention further provides methods of growth of a pathogen effecting the skin. Preferably, compo treating disease and/or disorders or symptoms thereof which sitions of the invention are used topically. For example, com comprise administering a therapeutically effective amount of positions of the invention can beformulated for the treatment a pharmaceutical composition comprising a compound of the or prevention of pathogen infections of the skin, including the formulae herein to a Subject (e.g., a mammal Such as a treatment of bacterial infections. The most common bacterial human). Thus, one embodiment is a method of treating a skin bacteria are Staphylococcus aureus and group A Subject Suffering from or Susceptible to an infectious disease b-hemolytic streptococci. Bacterial infections of the skin or disorder or symptom thereof. The method includes the step include infections by e.g., Mycobacterium tuberculosis, of administering to the mammal a therapeutic amount of a Micrococcus luteus, Propionibacterium acnes, Staphylococ compound herein (e.g., B-amyloid and analogs thereof) Suf cus aureus, Streptococcus pyogenes, and Staphylococcus epi ficient to treat the disease or disorder or symptom thereof, dermidis. Bacterial infections can result in a variety of skin under conditions such that the disease or disorder is treated. conditions, including impetigo, which is associated with S. 0077. The methods herein include administering to the aureus or S. pyogenes, erysipelas, which is associated with Subject (including a subject identified as in need of Such Streptococcus or Staphylococcus, and cellulitis, which is treatment) an effective amount of a compound described associated with S. aureus or S. pyogenes infection. Folliculi herein, or a composition described herein to produce Such tis, furuncles, and carbuncles are infections associated with S. effect. Identifying a subject in need of such treatment can be aureus. Compounds of the invention have natural antibiotic in the judgment of a Subject or a health care professional and and/or bacteriostatic activities, which are useful for the treat can be subjective (e.g. opinion) or objective (e.g. measurable ment or prevention of skin conditions associated with bacteria by a test or diagnostic method). or a bacterial infection. 0078. The therapeutic methods of the invention (which I0081 -amyloid peptides, oligomers, and analogs thereof include prophylactic treatment) in general comprise admin are typically present in a diluent or carrier at levels ranging istration of a therapeutically effective amount of the com from about 0.1% to about 95%. The methods herein contem pounds herein, Such as a compound of the formulae (e.g., plate administration of an effective amount of compound or B-amyloid, oligomers thereof, and analogs thereof) herein to compound composition comprising B-amyloid peptides, oli a subject (e.g., animal, human) in need thereof, including a gomers, and analogs thereof to achieve the desired or stated mammal, particularly a human. Such treatment will be suit antibiotic or bacteriostatic effect. Preferably, the amount of ably administered to Subjects, particularly humans, Suffering active ingredient is combined with carrier materials to form a from, having, Susceptible to, or at risk for a disease, disorder, composition (e.g., cream, lotion, powder, topical spray) Suit or symptom thereof. Determination of those subjects "at risk” able for application to the skin. For some applications, anti can be made by any objective or subjective determination by inflammatory, antibiotic or bacteriostatic compositions of the a diagnostic test or opinion of a subject or health care provider invention are formulated for topical application. (e.g., genetic test, enzyme or protein marker, Marker (as I0082. A typical anti-inflammatory, antibiotic or bacterio defined herein), family history, and the like). The compounds static formulation will contain from about 1% to about 95% herein may be also used in the treatment of any other disor B-amyloid peptides, oligomers, and analogs thereof, where ders in which an infectious pathogen or other microbe may be the bottom of the range is any integer between 5 and 94 and implicated. the top of the range is any integer between 6 and 95, where the 0079. In one embodiment, the invention provides a B-amyloid peptides, oligomers, and analogs thereof are pro method of monitoring treatment progress. The method vided in a carrier that is Suitable for topical application. includes the step of determining a level of diagnostic marker Where antibiotic or bacteriostatic compositions are desired, (Marker) (e.g., any target delineated herein modulated by a the compositions of the invention are preferably formulated compound herein, a protein or indicator thereof, etc.) or diag with a carrier suitable for topical administration. The ratio of nostic measurement (e.g., Screen, assay) in a Subject suffering B-amyloid peptides, oligomers, and analogs thereof to carrier from or Susceptible to a disorder or symptoms thereof asso ranges between about 1:2 and 1:100. Preferred ratios include ciated with a pathogen infection, in which the Subject has 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:20, 1:30, 1:50, been administered a therapeutic amount of a compound 1:75, and 1:100. Alternatively, compositions of the invention herein sufficient to treat the disease or symptoms thereof. The include at least about 1%, 10%, 20%, 30%, 50%, 60%, 75%, level of Marker determined in the method can be compared to 80%, 90%, or 95% B-amyloid peptides, oligomers, and ana known levels of Marker in either healthy normal controls or in logs thereof in a diluent or carrier. other afflicted patients to establish the subject’s disease sta I0083. In preferred embodiments, the preparation includes tus. In preferred embodiments, a second level of Marker in the between 1 and 95% (e.g., 1,2,3,4,5,6,7,8,9, 10, 25, 75,80, subject is determined at a time point later than the determi 90, or 95%) -amyloid peptides, oligomers, and analogs US 2012/0058935 A1 Mar. 8, 2012 thereof in a carrier or diluent. Alternatively. Such preparations radioprotective. Skin care compositions include topically contain from about 20% to about 80% f-amyloid peptides, applied over-the-counter compositions, anti-fungal treat oligomers, and analogs thereof. Compositions containing ments, anti-acne treatments, skin protectants, and antiperspi B-amyloid peptides, oligomers, and analogs thereof are rantS. manufactured by ordinary methods. B-amyloid peptides, oli I0087. The invention also provides methods for making gomers, and analogs thereof Suitable for addition to products personal care compositions comprising combining an effec can be formulated as ordinary tablets, capsules, Solids, liq tive amount of a B-amyloid peptides, oligomers, and analogs uids, emulsions, slurries, fine granules or powders, which are thereof composition of the invention with a physiologically Suitable for administration to products during their prepara acceptable carrier or excipient to provide a personal care tion, following preparation but prior to storage, or at any time composition. The type of carrier utilized in the present inven prior to their sale to a vendor or consumer. Lower or higher tion depends on the type of product form desired for the amounts than those recited above may be required. The com personal care composition. In some embodiments, the carrier positions delineated herein include the compounds of the is a solid, while in other embodiments, it is semi-solid or formulae delineated herein, as well as additional anti-inflam liquid. Suitable carriers include liquids, as well as semi-solids matory, antibiotic or bacteriostatic activity agents if present, (e.g., creams, lotions, gels, Sticks, ointments, pastes, sprays in amounts effective for inhibiting an inflammatory process, and mousses). In particular, carriers that are lotions, creams or bacterial growth or reducing bacterial proliferation. Compo gels are useful for the topical application of a combination of sitions of the invention are used to prevent, reduce, inhibit, the invention. The carrier itself may be inert or may possess slow or stabilize the growth, proliferation, or survival of a dermatological benefits of its own. Preferably, the carrier is bacterium (e.g., . . . . Micrococcus luteus, E. faecalis, Propi physically and chemically compatible with a B-amyloid pep Onibacterium acnes, Staphylococcus aureus, Streptococcus tides, oligomers, and analogs thereof composition of the pyogenes, and Staphylococcus epidermidis). invention described herein, and does not unduly impair sta 0084 Lower or higher doses than those recited herein may bility, efficacy or other use benefits associated with the com be required to effectively inhibit an epidermal bacterium positions of the present invention. (e.g.,..., Micrococcus luteus, E. faecalis, Propionibacterium I0088 Alternatively, the compounds of the invention are acnes, Staphylococcus aureus, Streptococcus pyogenes, and provided as oral compositions (e.g., tablets, capsules, liquids, Staphylococcus epidermidis). Specific dosage and treatment or Sublingual formulations). regimens are determined empirically as described herein. Compositions of the invention are also useful for preventing Wound Treatment the establishment of a skin condition, such as a skin bacterial infection and for maintaining or enhancing the health and/or 0089 Antimicrobials of the invention can be used alone or appearance of skin. in conjunction with an absorbent and/or adsorbent material for the treatment of wounds. In one embodiment, a composi Dermatologic Formulations tion comprising B-amyloid peptides, oligomers, and analogs thereof is used to treat a Surgical site prior to, during, or 0085 Compositions of the invention containing f -amy following Surgery. In other embodiments, a composition of loid peptides, oligomers, and analogs thereof are useful not the invention comprising B-amyloid peptides, oligomers, and only for promoting the health and condition of skin, but also analogs thereof is used to irrigate a wound or other site having for enhancing the appearance of skin. Accordingly, the inven a propensity to develop a pathogen infectionl. Materials com tion provides for cosmetic and dermatological preparations of prising antimicrobials of the invention are suitable for absorb various forms. Preferably, the present invention provides per ing fluids that are contaminated with or are Susceptible to Sonal care compositions that include an effective amount of a contamination by a microbe, Such as bodily fluids, secretions, combination of compounds of the invention, with the balance or excretions. Bodily fluids, secretions, or excretions include, of the composition comprising one or more compounds from but are not limited to, blood, urine, saliva, serous fluid, Syn the group of carriers, excipients, liposomes, active ingredi ovial fluid, gastric secretions, cerebrospinal fluid, Sweat, ents, biological or botanical products, humectants, emol tears, bile, chyme, mucous, vitreous humor, lymph, wound lients, surfactants, thickening agents, silicone components, exudate, feces, blood (e.g., menstrual blood), and semen. In organic Sunscreens, preservatives, neutralizing agents, per one embodiment, an antimicrobial composition of the inven fumes or pigments. Such compositions improve or enhance tion is incorporated into an absorbent fibrous or non-fibrous skin health, condition, or appearance, and are provided, for material Suitable for use as a wound dressing, a medical example, as a cream, lotion, liquid, Solution, an anhydrous sponge, a hemostatic article, a hemostatic article for the nose, preparation, an oil-free preparation, an emulsion or micro an adhesive bandage, a wound packing, an internal vascular emulsion, a gel, a Solid stick, a wipe, a patch, an ointment, or closure packing, an external vascular closure dressing, a as an aerosol. It may also be advantageous to administer swellable absorbent article, or a fibrotic wound packing compositions of the invention in encapsulated form, for article. example in collagen matrices or in other conventional encap Sulation materials, for example as cellulose encapsulations, in gelatine, wax matrices, or liposomally encapsulated. Methods for Assaying Antimicrobial Activity I0086. In one approach, compositions of the invention are 0090 Specific dosage and administration regimens are provided in a personal care composition, Such as a moistur determined empirically as described herein. Methods for izing body wash, body wash, antimicrobial cleanser, skin determining the anti-microbial activity of a composition of protective cream, body lotion, facial cream, moisturizing the invention are known in the art. In one embodiment, anti cream, facial cleansing emulsion, Surfactant-based facial microbial activity is estimated by determining the number of cleanser, facial exfoliating gel, anti-acne treatment, facial microbes that survive incubation with the candidate anti toner, exfoliating cream, facial mask, after-shave balm or microbial using an assay for colony forming units. Such US 2012/0058935 A1 Mar. 8, 2012

methods are described, for example, by Datta et al., Appl results in a concentration of the therapeutic that, combined Environ Microbiol. 1997 October; 63(10): 4123-4126, Rhee with other components, is effective in ameliorating, reducing, et al., Appl Environ Microbiol. 2003 May: 69(5): 2959-2963: or stabilizing a skin infection. The compound may be con or by determining the effect of an antimicrobial on a bacterial tained in any appropriate amount in any Suitable carrier Sub colony of L. monocytogenes, as measured in an inhibitory stance, and is generally present in an amount of 0.1-95% by halo assay (Dieuleveux et al., Appl Environ Microbiol. 1998 weight of the total weight of the composition. Preferably, the February; 64(2): 800-803). In one embodiment, a microscale composition is provided in a dosage form that is Suitable for assay is used (Barreteau et al., “A rapid method for determin topical administration. In other embodiments, the composi ing the antimicrobial activity of novel natural molecules. J tion is provided in a form that is suitable for oral administra Food Prot. 2004 September; 67(9): 1961-1964). tion. The pharmaceutical compositions may be formulated according to conventional pharmaceutical practice (see, e.g., Prophylactics and Therapeutics Remington: The Science and Practice of Pharmacy (20th ed.), 0091 For therapeutic uses, the B-amyloid peptides, oligo ed. A. R. Gennaro, Lippincott Williams & Wilkins, 2000 and mers, and analogs thereof disclosed herein may be adminis Encyclopedia of Pharmaceutical Technology, eds. J. Swar tered topically. Other routes of administration include, for brick and J. C. Boylan, 1988-1999, Marcel Dekker, New example, nasal, local, Subcutaneous, intramuscular, or intra York). dermal injections that provide continuous, Sustained levels of 0094 Pharmaceutical compositions according to the the drug in the patient. Treatment of human patients or other invention may be formulated to release the active compound animals will be carried out using a therapeutically effective Substantially immediately upon administration or at any pre amount of a combination therapeutic in a physiologically determined time or time period after administration. The lat acceptable carrier. Suitable carriers and their formulation are ter types of compositions are generally known as controlled described, for example, in Remington's Pharmaceutical Sci release formulations. For Some applications, controlled ences by E. W. Martin. The amount of the therapeutic agent to release formulations obviate the need for frequent dosing be administered varies depending upon the manner of admin during the day. Any of a number of strategies can be pursued istration, the age and body weight of the patient, and with the in order to obtain controlled release in which the rate of clinical symptoms of the skin condition. Generally, amounts release outweighs the rate of metabolism of the compound in will be in the range of those used for other agents used in the question. In one example, controlled release is obtained by treatment of skin conditions associated with acne, although in appropriate selection of various formulation parameters and certain instances lower amounts will be needed because of the ingredients, including, e.g., various types of controlled increased specificity of the compound. A compound is admin release compositions and coatings. Thus, the therapeutic is istered at a dosage that controls the clinical or physiological formulated with appropriate excipients into a pharmaceutical symptoms of a skin condition as determined by a diagnostic composition that, upon administration, releases the therapeu method known to one skilled in the art. tic in a controlled manner. Examples include single or mul 0092. Therapeutic compounds and therapeutic combina tiple unit tablet or capsule compositions, oil solutions, Sus tions are administered in an effective amount. In certain pensions, emulsions, microcapsules, microspheres, embodiments, compounds of the invention, such as those molecular complexes, nanoparticles, patches, and liposomes. described herein, are administered at dosage levels of about 0.1 ppm to about 3000 ppm. In particular embodiments, the B-amyloid peptides, oligomers, and analogs thereof are Topical Administration present at about 0.1, 0.2,0.5,0.75, 1,2,3,5,10,15, 20, 25, 50, 0.095 Topical administration of the pharmaceutical com 75, 100, 200,250, 300,350, 400, 500, 750, 1000, 1250, 1500, positions of this invention is especially useful for preventing 2000, 2500, and 3000 ppm. Other compounds of the invention or treating a skin disease or disorder or for enhancing the are administered in an amount of about 0.0001 to 4.0 grams appearance of skin. For application topically to the skin, the once per day (or multiple doses per day in divided doses) for pharmaceutical composition should be formulated with a adults. Thus, in certain embodiments of this invention, a Suitable ointment containing the active components Sus compound herein is administered at a dosage of any dosage pended or dissolved in a carrier. Carriers for topical admin range in which the low end of the range is any amount istration of the compounds of this invention include, but are between 0.1 mg/day and 400 mg/day and the upper end of the not limited to, mineral oil, liquid petroleum, white petroleum, range is any amount between 1 mg/day and 4000 mg/day propylene glycol, polyoxyethylene polyoxypropylene com (e.g., 5 mg/day and 100 mg/day, 150 mg/day and 500 mg/day, pound, emulsifying wax and water. Alternatively, the phar 300 mg/day-1000 mg/d (oral)). In other embodiments, a com maceutical composition can be formulated with a suitable pound herein, is administered at a dosage range in which the lotion or cream containing the active compound Suspended or low end of the range is any amount between 0.1 ppm/day and dissolved in a carrier. Suitable carriers include, but are not 4999 ppm/day and the upper end of the range is any amount limited to, mineral oil, Sorbitan monostearate, polysorbate 60, between 0.2 ppm/day and 5000 ppm/day. Preferably, a com cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl bination of the invention is administered topically once per alcohol and water. day in a composition comprising at least about 1% B-amyloid peptides, oligomers, and analogs thereof. The dosing interval Therapeutic Combinations can be adjusted according to the needs of individual patients. For longer intervals of administration, extended release for 0096. If desired, compositions comprising B-amyloid pep mulations can be used. tides, oligomers, and analogs thereofare administered alone or in combination with other therapeutics commonly used for Formulation of Pharmaceutical Compositions the treatment of skin infections. In one embodiment, a com 0093. The administration of a compound for the treatment position of the invention comprises one or more of doxycy of a skin disease or disorder may be by any suitable means that cline, tetracycline, erythromicin, minocycline, trimethoprim, US 2012/0058935 A1 Mar. 8, 2012

azithromycin, and/or clindamycin. If desired a composition following: sink, tile, bathtub, shower wall, toilet bowl, of the invention is administered together with an inert carrier kitchen countertop, tabletop, table covering, cutting board, Suitable for topical administration, including but not limited eating utensil, stove top, oven, microwave oven, refrigerator, to, cocamidopropyl betaine, methylparaben, butylene glycol, wall, floor, and/or window. Such surfaces are commonly benzoic acid, octoxynol-9, and/or urea. found in households, hospitals, and food production facili ties. In addition, the compositions can be used on the interior Subject Monitoring and exterior Surfaces of common objects of construction, 0097. The disease state or treatment of a subject having a including, but not limited to exterior and interior surfaces of skin disease or disorder can be monitored during treatment an airplane, automobile, bathtub, boat, building, fluid distrib with a composition of the invention. Such monitoring may be uting system, household-appliance, household fixture, useful, for example, in assessing the efficacy of a particular shower stall, sink, ship, sanitary closet, vehicle, water distri agentinapatient. therapeutics that promote skinhealth or that bution system, water recirculation system, and/or combina enhance the appearance of skin are taken as particularly use tions thereof, and further including the finished, laminated, ful in the invention. coated and/or painted Surfaces thereof. 0101 If desired, compositions of the invention used as Cleaning Compositions cleaners or sanitizers are provided in combination with an 0098 Anti-microbial compositions containing B-amyloid effective amount of one or more surfactants. Such surfactants peptides, oligomers, and analogs thereof may be employed in include, but are not limited to, nonionic, semi-polar, anionic, the form of aqueous or non-aqueous dilutions for application cationic, Zwitterionic, and/or amphoteric Surfactants. In one and used to sanitize a Surface. By 'sanitize' is meant prevent aspect of this embodiment, the Surfactant includes, but is not or treat microbial contamination of the surface. Desirably, limited to, lauryl Sulfate, laurylether Sulfate, cocamidopropy compositions of the invention reduce microbial contaminants 1betaine, alkyl polyglycosides, and/or amine oxides. The Sur in the inanimate environment to levels considered safe factant content in and/or used in combination with the according to public health ordinance. An anti-microbial of the improved cleaning composition is about 0.1-2 weight per invention may formulated as an aerosol, mist, spray, foam, cent. In yet a further aspect of this embodiment, the surfactant liquid, wash, rinse, or as a bath to treat Submerged items, content in and/or used in combination with the improved articles or Surfaces, and/or added to an aqueous system to cleaning composition is about 0.15-1.5 weight percent Instill treat submerged surfaces. Thus, a wide variety of suitable yet a further aspect of this embodiment, the surfactant content application methods include for example, but are not limited to, pouring, spraying, application with a trigger sprayer, aero in and/or used in combination with the improved cleaning Sol sprayer or device containing a pressurized propellant and/ composition is about 0.2-1.5 weight percent. In another or condensed gas, spraying onto a surface from a container aspect of this embodiment, the Surfactant content in and/or attached to a hose, wiping onto a surface with a pre-moistened used in combination with the improved cleaning composition disposable device Such as for example, but not limited to, a is about 0.2-1.25 weight percent. In yet another aspect of this nonwoven wipe, cloth and/or sponge wetted with the inven embodiment, the surfactant contenti is about 0.5-1.25 weight tive compositions. Suitable application methods include any percent. In still another aspect of this embodiment, the Sur method in which the inventive compositions are applied factant content is about 0.1-1 weight percent. In still yet directly in either neatform, or concurrent with and/or follow another aspect of this embodiment, the Surfactant content is ing dilution of a concentrated composition with a suitable about 0.15-0.8 weight percent. In a further aspect of this aqueous diluent, such as for example water. embodiment, the surfactant content is about 0.2-0.4 weight 0099. In one representative embodiment, B-amyloid pep percent. In yet a further aspect of this embodiment, the sur tides, oligomers, and analogs thereof are formulated as con factant content is less than about 0.5 weight percent. centrated mixtures of ingredients, optionally including a first 0102 The cleaning or sanitizing composition typically aqueous or non-aqueous diluent (e.g., ethanol) that may be includes water. When the improved cleaning composition is a further diluted to prepare a ready-to-use solution. During use, liquid, water based, ready-to-use cleaner, the water content of and in order to sanitize or disinfect a surface, the inventive the improved cleaning composition is generally over 50 compositions are applied to and allowed to contact the Surface weight percent of the improved cleaning composition. Typi for a proscribed time to effect microbial reduction or kill. cally, the liquid ready-to-use improved cleaning composition After this contact time, the inventive compositions may be includes at least about 80 weight percent water; however, allowed to remain in place, or may optionally be wiped or higher or lower water contents can be used. removed from the surface by some suitable means. Option ally, the treated surface can be rinsed with water to remove the 0103) One or more additional anti-microbial compounds inventive compositions following treatment. Compositions of can be included in and/or used in combination with the the invention may optionally include cleaning agents and improved cleaning composition to enhance the biocidal effi other adjuncts and hence provide simultaneous cleaning and cacy of the cleaning composition. Such anti-microbial com antimicrobial treatment of Surfaces. pounds include, but are not limited to, glycine, Sodium 0100. The antimicrobial compositions can be used in a actetate, Sorbic acid, Sodium dehydroacetate, Sodium lactate, household, commercial, restaurant, medical, business and/or Sodium benzoate, p-hydroxy benzoate, e-polylysine, milt outdoor environment. Surfaces to which the antimicrobial protein, lysozyme, or any plant derived antimicrobial com composition may be applied include, but are not limited to ponents, alcohols, peroxides, boric acid and borates, chlori those made from metal, plastic, Stone, glass, ceramic, painted nated hydrocarbons, organometallics, halogen-releasing Surfaces, wallpaper, textiles, carpets, and the like. Antimicro compounds, mercury compounds, metallic salts, pine oil, bials of the invention may be used to sanitize any of the essential oils, organic Sulfur compounds, iodine compounds, US 2012/0058935 A1 Mar. 8, 2012

silver nitrate and other silver compounds, quaternary phos ing, and therapeutic methods of the invention, and are not phate compounds, and/or phenolics. intended to limit the scope of what the inventors regard as their invention. Kits 0104. The invention provides kits for the treatment or pre EXAMPLES vention of a skin disease or disorder, or symptoms thereof. In one embodiment, the kit includes a pharmaceutical pack com Example 1 prising an effective amount of a 3-amyloid peptides, oligo mers, and analogs thereof. Preferably, the compositions are A? Peptides Possess Antimicrobial Activity present in unit dosage form. In some embodiments, the kit comprises a sterile container which contains a therapeutic or 0109 Antimicrobial activity against a particular microor prophylactic composition. Such containers can be boxes, ganism is measured in vitro by a peptide's minimal inhibitory ampules, bottles, vials, tubes, bags, pouches, blister-packs, or concentration (MIC), which is defined as the lowest concen other suitable container forms known in the art. Such con tration able to visibly inhibit growth overnight. The MICs of tainers can be made of plastic, glass, laminated paper, metal synthetic LL-37. Af40, and A342, against a panel of clini foil, or other materials suitable for holding medicaments. cally relevant organisms (Table 1). 0105. If desired compositions of the invention or combi nations thereof are provided together with instructions for TABLE 1 administering them to a subject having or at risk of develop Table 1: The antimicrobial activity of synthetic AB1-42 ing a skin disease or disorder. The instructions will generally (AB42), AB1-40 (AB40), LL-37 (LL-37), reverse include information about the use of the compounds for the AB42-1 (rAB42), or scrambled AB42 (scAf42) treatment or prevention of a skin disease or disorder. In other peptides were determined as minimal inhibitory concentrations embodiments, the instructions include at least one of the MIC) against 12 microorganisms. following: description of the compound or combination of MIC ml compounds; dosage schedule and administration for treat ment of a skin condition associated with acne, dermatitis, Organism AB42 AB40 roAB42 LL-37 reAB42 scAB42 psoriasis, or any other skin condition characterized by inflam Candida albicans O.78 O.78 O.78 6.25 >25 >50 mation or a bacterial infection, or symptoms thereof; precau Escherichia coi 1.56 1S6 3.13 1.56 >50 >50 Staphylococcus 3.13 SO 3.13 25 >50 >50 tions; warnings; indications; counter-indications; overdosage epidermidis information; adverse reactions; animal pharmacology; clini Streptococci is 6.25 12.5 6.25 1.56 50 >50 cal studies; and/or references. The instructions may be pneumoniae printed directly on the container (when present), or as a label Staphylococcits 6.25 2S 12.5 6.25 >50 >50 (iiietiS applied to the container, or as a separate sheet, pamphlet, Listeria 6.25 2S 6.25 2S >50 50 card, or folder supplied in or with the container. monocytogenes 0106 The recitation of a listing of chemical groups in any Enterococcits 6.25 SO 3.13 6.25 50 >50 definition of a variable herein includes definitions of that faecalis Streptococci is 12.5 50 >50 12.5 >50 >50 variable as any single group or combination of listed groups. agalaciae The recitation of an embodiment for a variable or aspect Pseudomonas >50 >50 >50 6.25 >50 >50 herein includes that embodiment as any single embodiment aeruginosa or in combination with any other embodiments or portions Streptococci is >50 >50 >50 6.25 >50 >50 thereof. pyogeneS Streptococci is >50 50 >50 6.25 >50 >50 0107 The practice of the present invention employs, mitis unless otherwise indicated, conventional techniques of Streptococci is >50 >50 >50 50 >50 >50 molecular biology (including recombinant techniques), Saivarius microbiology, cell biology, biochemistry and immunology, Antimicrobial activity was assayed by broth microdilution susceptibility test on 96-well which are well within the purview of the skilled artisan. Such plates with microbial growth in wells determined by visual inspection following an over night incubation. Inhibition of growth in plate wells was confirmed by alamar blue cell techniques are explained fully in the literature. Such as, viability assay and by surface plating of incubants on agar and counting CFU. Inoculums “Molecular Cloning: A Laboratory Manual, second edition contained mid-logarithmic phase cells, Consistent with antimicrobial activity specific to the (Sambrook, 1989); “Oligonucleotide Synthesis” (Gait, AB sequence, inhibition was not observed for reverse and scrambled peptides. 1984): “Animal Cell Culture” (Freshney, 1987); “Methods in 0110. The antimicrobial activity of A? peptides was Enzymology” “Handbook of Experimental Immunology’ equivalent to or greater than LL-37 for seven of the pathogens (Weir, 1996); “Gene Transfer Vectors for Mammalian Cells” tested. These data indicate that AB is a bona fide AMP with (Miller and Calos, 1987); “Current Protocols in Molecular potencies similar to, or, in Some cases surpassing those of Biology” (Ausubel, 1987); “PCR: The Polymerase Chain LL-37. The synthetic AB peptides demonstrated antibiotic Reaction. (Mullis, 1994): “Current Protocols in Immunol activity against Gram-negative and Gram-positive bacteria ogy” (Coligan, 1991). These techniques are applicable to the and the yeast C. albicans. Activity was isoform-specific for production of the polynucleotides and polypeptides of the six organisms with AB42 showing greater potency compared invention, and, as such, may be considered in making and to AB40. Equivalent findings were observed for recombinant practicing the invention. Particularly useful techniques for Af342, material that is free of the potentially toxic contami particular embodiments will be discussed in the sections that nants associated with conventional Solid-phase peptide Syn follow. thesis. Rodent AB42 also demonstrated antimicrobial activ 0108. The following examples are put forth so as to pro ity. However, microbial growth was not inhibited by reverse vide those of ordinary skill in the art with a complete disclo (raf342) or scrambled (scAB42) negative control peptides, Sure and description of how to make and use the assay, Screen thus confirming the antimicrobial action is peptide-specific. US 2012/0058935 A1 Mar. 8, 2012

0111 AMPs, including LL-3721, can be bacteriostatic ciation has been shown for LL-37 and the disorder morbus or bactericidal depending on peptide concentration, ionic Kostmann in which patients deficient in this AMP cannot strength, and the type of stressor a colony has previously mount an effective defense against pathogens 32. A rela encountered. The growth curves for E. faecalis in the pres tionship between human immunodeficiency and low AB lev ence of AB42 suggest a predominantly bacteriostatic action els has not been investigated. However, knockout mice that for the peptide against this organism under our incubation lack the proteases that generate endogenous rodent AB appear conditions (FIG. 1). Consistent with previous studies, LL-37 to have increased susceptibility to pathogens 33. BACE1 showed potent bactericidal activity against E. faecalis. knockout (KO) mice that generate low levels of AB and Microbial growth resumed at later time points, most likely BACE1- and BACE2-deficient double KO mice, which do not due to degradation of LL-37 and AB by protective bacterial express AB, have mortality rates of 40 and 60 percent, respec proteases (FIG. 1). tively. Housing the animals in a pathogen-free environment 0112 The capacity to associate with microbial lipid bilay restores survival rates to that of wild-type mice (>95 percent). ers is considered a definitive feature of AMPs 22. Most The etiology of the immunodeficiency has been investigated antimicrobial peptides are cationic to facilitate binding to but not identified. Adaptive immune responses to vesicular anionic bacterial membranes. However, AB peptides are stomatitis virus are the same for BACE-KO and wild-type anionic under physiological conditions 23. Nonetheless, mice. In addition, markers for adaptive immune system func data from light microscopic examination of immunostained tion are normal in BACE-KO mice, including leukocytes bacteria pre-incubated with A? confirm that the peptide binds migration into the peritoeum following thioglycolateacute to the surface of bacterial cells (FIG. 2). Binding of AB to induced acute peritonitis and T-cell cytotoxiocity towards bacterial membranes is consistent with previous studies non-host cells. More recently, in a clinical trial of the AB42 showing that AB readily binds and disrupts negatively lowering agent tarenflurbil patients receiving the drug have charged synthetic lipid bilayers 24.25 and anionic mito significantly increased rates of infection 34. Increased chondria membranes 26.27.28, believed to have been origi pathogen Susceptibility of apparently adaptive immunocom nally derived from bacterial membranes. petent BACE-KO mice and AD patients with suppressed AB 0113. The next experiments tested whether the antimicro expression is consistent with our finding that AB may have a bial activity observed for synthetic peptides in vitro could be normal protective function as an antimicrobial peptide of the identified in temporal lobe and cerebellum from human brain. innate immune system. Typically B-amyloid load is high in AD temporal lobe and low 0115 The immunostatus of APP knockout (APP-KO) in cerebellum. Tissue taken from AD (n=32) or age matched mice has yet to be characterized. APP is a member of a larger control Subjects (n=13) were homogenized and normalized protein family that includes the amyloid protein precursor for protein. A?40 and A?42 levels in brain homogenates were like proteins 1 and 2 (APLP1 and APLP2) 35,36). APP and determined by ELISA. Homogenates were then diluted into APLP proteins appear to have overlapping and partially culture broth and inoculated with Candida albicans. Growth redundant functions 37.38.39 and share processing path of C. albicans was determined using a fluorescence-based ways, including BACE-mediated generation of APLP-de alamar blue microplate assay previously described for fol rived peptides analogous to AB 37.38.39.40. It is unclear to lowing cell viability with this organism 29. AD temporal what degree APLP-derived proteins may compensate for lobe homogenates inhibited the growth of C. albicans signifi deficiencies associated with low APP expression. Mice lack cantly more (p=0.0048) than non-demented control samples ing both APP and APLP proteins (triple APP/APLP KO (FIG. 3A). Consistent with an AB-mediated antimicrobial mice) show early postnatal mortality with severe develop activity in AD temporal lobe homogenates, a significant dif mental abnormalities 41,42,43.44.45. ference in C. albicans growth was not observed with cerebel 0116 Interestingly, local cortical dysplasias can be infec lum samples, which carry a considerably lower AB load. Also tion-mediated and are observed in 68% of triple APP/APLP consistent with Af-mediated antimicrobial activity, C. albi KO-mice 45. Partial penetrance is also suggestive of an cans growth significantly correlated with A? concentration in environmental component in this ectopia. temporal lobe homogenates (FIG. 3B), but not in cerebellum 0117 Recent studies have shown that while the adaptive samples with Pearson's correlation coefficients (r) of -0.484. immune system has limited access to the brain, the CNS can p=0.0012 and -0.091, p=0.56, respectively. In addition, the still mount a robust response to invading pathogens via anti increased antimicrobial activity of AD temporal lobe samples microbial peptides and the innate immune system. Numerous could be significantly attenuated (p=0.0007) by immun innate immune molecules with potent antimicrobial activity odepletion of homogenates with anti-AB antibodies (FIG. are found in brain, including the recently identified chromog 4A), consistent with an AB-mediated antimicrobial activity in ranins 46, neuropeptides neurokinin-1, enkelytin and pep AD brain. Analysis of immunodepleted homogenates con tide B, neuropeptide Y. polypeptide tyrosine-tyrosine, and the firmed AB levels were attenuated in samples incubated with peptide hormones C.-melanocyte stimulating hormone, rabbit anti-A? antibody (FIG. 4B). Additional experiments adenoregulin, adrenomedullin and proadrenomudullin, corti confirmed that antimicrobial activity in AD temporal lobe costatin RK-1, neurotensin, and bradykinin 47. Consistent homogenates is also attenuated following immunodepletion with an antimicrobial role for brain generated AB, AD tem with the anti-A? mouse monoclonal antibody 6E 10 (FIG. 5). poral lobe homogenates contained an average of 24% greater 0114 AB peptides inhibited the growth of eight of 12 activity against C. albicans than samples from non-AD Sub clinically important pathogens Screened (Table 2), including jects (FIG. 3A). Furthermore, higher AB levels in temporal the bacteria S. pneumoniae, which is a leading cause of bac lobe samples correlated with increased inhibition of C. albi terial meningitis 30, and C. albicans, the most common cans (FIG.3B) while immunodepletion of AB from AD brain cause of neurocandidiasis 31. If the normal function of AB homogenates restored antimicrobial activity to levels equiva is to function as an AMP, then an absence of the peptide may lent to those of control homogenates (FIGS. 4A and 5). result in increased vulnerability to infection. Such an asso Immunoblot analysis confirmed attenuated AB levels in anti US 2012/0058935 A1 Mar. 8, 2012

AB antibody immunodepleted samples and low B-amyloid ciation with AD was a homologue of CD33, a lectin involved load in cerebellum tissue (FIG. 4B). These data support a in the innate immune system 70. protective role for AB under the conditions found in the brain I0120 While dozens of diseases have been suggested to milieu even though in vivo concentrations of soluble peptide involve immune abnormalities, for most, the underlying are Substantially lower than levels in experiments using Syn cause of the aberrant immunoresponse remains unclear. For thetic peptide 48). Several factors may contribute to this AD, traumatic brain injury 71, stroke 72 and certain forms apparent discrepancy. First, Synergistic AMP interactions in of inhalant anesthetics 73 have been linked to increased vivo potentiate antimicrobial activity 49. This effect has cerebral AB levels. While an infection-mediated pathological been demonstrated for CRAMP (rodent LL-37), for which mechanism for AD is one possibility, this remains to be inves peptide levels in rodent CNS do not approach concentrations tigated, and other non-microbial factors may also be involved. that are needed to obtain positive signals in in vitro assays. Interestingly, peptides containing the microtubule binding However, rat brain extracts depleted of CRAMP have sub sites on tau proteins have also been shown to harbor antimi stantially attenuated antimicrobial activity 50. Moreover, crobial properties 74. mutant mice lacking CRAMP are more susceptible to CNS I0121 The capacity to associate with lipid bilayers is con infection by meningococcal meningitis 11. Second, AD sidered a definitive feature of AMPs, and the peptides usually affect their antimicrobial activity by membrane permeabili brain contains a large pool of neurotoxic oligomeric A? spe zation 75. Membrane disruption is also thought to be a cies 51.52. Oligomerization plays a key role in the targeting mechanism for AB-mediated cytotoxicity 24.26. The find and permeabilization of bacterial membranes by AMPs 19, ing that bacterial membranes stain positive for AB following 53.54. Neurotoxic oligomeric AB species present in AD incubation with the peptide (FIG. 2) is consistent with a brain may enhance the antimicrobial activity of homogenates mechanism that involves association with microbial lipid beyond that predicted from in vitro experiments, which add bilayers. While most AMPs are cationic, AB peptides are synthetic monomeric peptides to microbial cultures. anionic. Repulsive electrostatic forces between anionic pep 0118. A large body of data supports a central role for tides and electronegative phospholipids in bacterial mem neuroinflammation in AD neuropathology 55. A number of branes potentially limit antimicrobial activity of this class of studies have proposed AB as the Source of AD-associated AMP. However, in addition to our data, previous studies have inflammation 56. However, a re-evaluation of the role of AB conclusively shown that AB readily binds and disrupts both in inflammation may now be warranted in view of these data synthetic anionic lipid bilayers 24 as well as mitochondrial suggesting that the peptide functions as an AMP in tissues. membranes 26. Interestingly, mitochondria are thought to Inflammatory response in the immunologically privileged be of endosymbiont origin and have anionic membranes that CNS is mediated by the innate immune system. Rather than resemble the lipid bilayers of bacteria. A number of AMPs, AB acting as a sole independent initiator of neuroinflamma including LL-37, appear to target and disrupt the mitochon tion, our data raise the possibility that the peptide may be part drial membranes of parasitic protozoans 8. Recent studies of a response mounted by the innate immune system. Thus, have also identified a number of anionic mammalian peptides A? may be one of a family of AMPs known to contribute with antimicrobial activity, including CNS neuropeptides pro-inflammatory activities under disease conditions. At least 76 and peptide hormones 47. Structural studies on the one other disease has been shown to involve deposition of an important epithelial anionic AMP dermicidin have shown that AMP as amyloid, corneal amyloidosis. In corneal amyloido an overall positive charge is not a prerequisite for binding of sis the widespread and well-characterized antimicrobial pro bacterial membranes 77. Rather, the key modulators of lipid tein lactoferrin accumulates in the subepithelium as insoluble bilayers/peptide association are the peptides charge distribu amyloid 57.58. Semenogelin-derived antimicrobial pep tion and secondary conformation. Collectively, these data tides 59 are also deposited as seminal vesicle amyloid 60 indicate that AMP activity is not limited to cationic species in a common Sub-clinical pathology found in elderly men and that anionic peptides Such as AB can readily bind bacterial 61. membranes and act as potent antimicrobial agents. 0119) A number of studies have reported that the CNS of I0122. In E. faecalis cultures, AB was more resistant to AD patients is infected with pathogens including Chlamydia bacterial-mediated degradation than LL-37 (FIG. 1). Bacte pneumoniae (62. Borrelia spirochetes 63, Helicobacter rial defense mechanisms secrete proteases that target posi pylori (64), and HSV65). Deposition of B-amyloid has also tively charged peptides. Anionic AMPs are believed to be, at been reported for acquired immunodeficiency syndrome least in part, a host counter measure to bacterial resistance patients with brain HIV infection 66. Given the known mechanisms 78. Oligomerization is also thought to protect genetic influence on AB accumulation, genetic factors may AMPs from microbially-mediated degradation, and AB oli contribute to activation of the innate immune system by regu gomers have been shown to be highly protease resistant. An lating AB production and clearance. At one of the end of the anionic charge and propensity to oligomerize may therefore spectrum of known AD genes, highly penetrant mutations help render AB resistant to bacterial attack. such as those in the early-onset familial AD genes, APP, I0123 AMPs cytotoxicity is usually highly specific for PSENI, and PSEN2, would constitutively trigger cerebral AB microbes. However, AMPs can also be cytotoxic to select host accumulation with no need for activation of the innate cells under physiological conditions. Host cell cytotoxicity immune system 67. At the other end of the spectrum, con has been shown for LL-3714 which, like AB 79, is cyto sistent with the increase risk of AD associated with the e4 toxic towards vascular smooth muscle cells. AMP host cell variant of the apolipoprotein E gene 68, carriers of the e4 cytoxicity often involves disruption of mitochondrial func allele are reported to have higher rates of CNS infection for tion, an activity reported for both LL-37 80.81 and AB 27. several of these pathogens I69. Finally, in a recent family Thus, neurotoxicity that has been shown for AB is consistent based genome-wide association scan for late-onset AD, one with AMP behavior. The role of AMP host cell cytoxicity in of four genes achieving genome-wide significance for asso disease and defense is unclear. LL-37 cytotoxicity has been US 2012/0058935 A1 Mar. 8, 2012 implicated in disease pathology 14, but may also have a 29212, Streptococcus agalactiae ATCC 12386, Pseudomo normal function in antibody-dependent cell cytotoxicity, a nas aeruginosa ATCC 27853, Streptococcus pyogenes ATCC host mechanism for the clearance of virus-infected and trans 19615, Streptococcus mitis ATCC 6249, and Streptococcus formed cells 81. At present AB's host cell cytotoxicity is salivarius ATCC 13419. Bacteria were cultured aerobically in only associated with disease. Identification of AB as an AMP Mueller-Hinton broth (MHB), Brain and Heart Infusion broth raises the possibility that host cell cytotoxicity, or at least a (BHIB), or BHIB supplemented with 1% lysed horse blood component of this activity, may also have a role in innate and plated on Tryptone Soy Agar (TSA) plates containing 5% immunity. defibrinated sheep blood. C. albicans was grown in RPMI 0.124. In summary, the finding that AB is an antimicrobial 1640 medium (Hyclone, Logan, Utah) with 2% glucose buff peptide is the first evidence that the species responsible for ered (pH 7.0) and 0.165 M MOPS and surface plated on amyloidosis may have a normal function. This stands in Stark sabouraud dextrose agar plates. Culture conditions for each contrast to current models, which assume 3-amyloid deposi organism are included in Table 2. Organisms were subcul tion to be an accidental process resulting from the abnormal tured for 2 hrs to generate mid-logarithmic growth cultures behavior of an incidental product of catabolism. These data for use as inoculates in experiments. Media reagents were Suggest that increased AB generation, and resulting AD obtained from Becton, Dickinson and Company (Sparks, pathology, may be a mediated by a response of the innate Md.). immune system to a perceived infection. This model has important implications for current and future AD treatment TABLE 2 strategies. First, it raises the possibility of preventing amyloi dosis from initiating by pre-emptive targeting of pathogens/ Gram ATCC Growth Incub. insults that stimulate the brain's innate immune system. Sec Organism Stain No. Media (hrs) ond, our model identifies the inflammatory pathways of the Staphylococcusatiretts -- 25923 MHB 12 innate immune system as targets for modulating A3 genera Escherichia coi 25922 BHIB 12 Listeria monocytogenes -- 19112 BEHIB 18 tion/accumulation. The target pathways implicated here are Pseudomonas aeruginosa 27853 BEHIB 18 downstream of the inflammatory trigger. Thus, this approach Enterococci is faecalis -- 29212 BEHIB 18 would likely be useful independently of the involvement of Staphylococci is epidermis -- 12228 BHIB 18 Streptococci is agaiaciae -- 12386 BHIB LHB 12 infectious agents in AD pathology. Streptococci is pneumoniae -- 49619 BHIBALHB 12 0.125. The results reported herein were obtained using the Streptococci is mitis -- 6249 BEHIBFLHB 12 following methods and materials. Streptococcus pyogenes -- 1961S BHIB LHB 18 Streptococci is Saivarius -- 13419 BHIB LHB 12 Synthetic Peptides Candida albicans 10231 RPMI-1640 18 0126 Experiments used AB1-40 (AB40), AB1-42 (AB42), scrambled AB (scAB42), AB42-1 (raf342), LL-37, and Preparation of Inoculum Containing Mid-Logarithmic Phase scrambled LL-37 (scLL-37) peptides. AB and LL-37 peptides Cells were prepared and purified by Dr. James I. Elliott at Yale University (New Haven, Conn.) using solid-phase peptide I0129. Colonies from agar were transferred by sterile loop synthesis. Scrambled LL-37 peptide was from AnaSpec (San to growth media and incubated for 2 hrs at 37°C. to achieve Jose, Calif.). Recombinant human AB42 (recAf342) and a McFarland density of 0.5. Bacteria inoculum cell densities rodent AB42 (roAB42) were purchased from rPeptide (Bog were normalized to 5x10 cells/ml immediately before use art, Ga.) and Calbiochem (Gibbstown, N.J.) respectively. photometrically and Subsequently confirmed by colony Findings for recombinant and SPPS prepared peptides were count. Inoculum of C. albicans contained a cell density of equivalent in all experiments. 2.5x10 CFU/ml. Brain Samples Peptide Pre-Treatment and Preparation of Stock Solutions I0130 Bulk powdered peptides were first dissolved in 30% 0127 Human brains were obtained 12-24 hrs postmortem. trifluoroethanol (TFE) at 1 mg/ml. Five hundred microliter At the time of autopsy, one cerebral hemisphere was sec aliquots of the stock solutions were lyophilized and stored tioned and frozen at -70° C. and the other hemisphere was under nitrogen at -20° C. Stock solutions at 2 mg/ml were fixed in formalin for histological examination. The clinical prepared the day of experimentation from the peptide films by diagnosis of AD was confirmed by Subsequent histological solubilizing a second time in either water or 20% TFE. A? evidence of amyloid plaques and neurofibrillary tangles. stocks prepared in water were Sonicated and insoluble peptide Samples were provided by the Neurobiology Tissue Bank at aggregates pelleted by centrifugation (10 minx 16,000 g). the Mass General Institute for Neurodegenerative Disease Peptide concentrations in stock solutions were determined and Massachusetts General Hospital and included temporal immediately before use by bicinchoninic acid (BCA) protein lobe and cerebellum from 32 AD patients and 13 non-de assay. The validity of BCA for assaying AB peptides has been mented age-matched control Subjects. established previously 82. For MIC experiments, peptides were serially diluted into growth media. For other experi Cell Cultures ments stocks were diluted into required working buffers. 0128 Bacteria were from the AmericanType Culture Col Experiments included controls for peptide buffer vehicle lection (ATCC, Manassas, Va.) and included Candida albi alone. cans ATCC 10231, Escherichia coli ATCC 25922, Staphylo coccus epidermidis ATCC 12228, Streptococcus pneumoniae MIC Determination ATCC 496.19, Staphylococcus aureus ATCC 25923, Listeria I0131 Peptide antimicrobial activity was determined as monocytogenes ATCC 19112, Enterococcus faecalis ATCC minimal inhibitory concentration (MIC) 83. Experiments US 2012/0058935 A1 Mar. 8, 2012 identified peptide MIC by broth microdilution susceptibility three volumes of 10 mM phosphate buffer, pH 7.4 by 12 test in conjunction with CFU and alamar blue assays. Inocu passes in a glass-on-Teflon homogenizer. Homogenates were lum containing mid-logarithmic phase cells was dispensed diluted into RPMI-1640 media to inhibit C. albicans growth into the wells of polypropylene 96-well plates (Fisher, Pitts by approximately 50 percent (FIG. 5). Samples were then burgh, Pa.) containing seven two-fold dilutions of test peptide inoculated (2.5x10 CFU/ml) with mid-logarithmic growth in growth media. Plates were then incubated aerobically over culture of C. albicans and incubated aerobically for 3 hrs at night (12 to 18 hrs) at 37° C. Peptide MIC was taken as the 37° C. in 96-well microplates (100 ul?well). Alamar blue lowest concentration able reduce cell growth by CFU and reagent was added to wells (10 ul) and fluorescence measured alamar blue assays by at least two-fold and which correlated after 30 and 60 minutes incubation. Signal from test wells was with the visible loss of a growth button on the bottom of blanked on samples incubanted without C. albicans. Signal microtiter wells. Experiments were repeated a minimum of from homogenate blanks was equivalent to uninoculated three times for each organism, and tests included at least three media alone. Samples were assayed in quadruplicate. replicates for each assay condition. Experiments included control serial dilutions of buffer vehicle alone. Assaying A3 in Tissue Homogenates 0132) Note on radial diffusion assays (RDAs). RDAs have 0.136 AB40 and AB42 in samples were determined using been widely used in previous studies to assess AMP antimi commercially available ELISA kits (Covance, Princeton, crobial activity. However, in our experiments RDAs proved N.J.). Brain homogenates were assayed according to the unreliable for testing AB antimicrobial activity because the peptide failed to diffuse away from the point of application. manufacturer's instructions. AB solutions are prone to aggregation, particularly in the Immunodepletion of Brain Homogenates presence of eventrace amounts of metal, and interaction with contaminates or the media matrix may lead to rapid precipi 0.137 MagnaBind goat anti-rabbit IgG beads (Pierce, Ill.) tation of the peptide within the agar. AB peptides also appear were pre-incubated overnight with the AB specific rabbitanti to irreversibly absorb to the cellulose disks often used as amyloid f-peptide antibody (Invitrogen, CA) or rabbit IgG sample reservoirs in RDAs. then washed repeatedly. Pooled samples were prepared from CFU assay; Serial dilutions of incubants were prepared and temporal lobe (30 AD and 12 non-AD) or cerebellum (32 AD streaked onto the Surface of agar. The agar plates were then and 13 non-AD) homogenates. The pooled brain homoge incubated overnight at 37° C. and colonies forming units nates were incubated alone or with the antibody loaded beads counted. at 4°C. for 2 hrs. Final incubation conditions were 5 lug of antibody per mg of original tissue (w/w). Beads were pelleted Alamar Blue Cell Viability Assay and soluble fraction removed. Fractions were immunoblotted 0.133 Microbial growth was determined by following the and probed with the AB specific mAb 4G8 (Covance, Princ reduction of the synthetic metabolic substrate resaZurin eton, N.J.). Analysis confirmed anti-A? antibody treated (alamarblue) to a fluorescent product by respiratory enzymes homogenates were depleted of AB (FIG. 4B). Soluble frac in living cells 84. Alamarblue assay is used in high through tions were then tested for antimicrobial activity against C. put Screens for antimicrobial agents 85 and is available albicans by alamar blue assay. commercially in kit form from Invitrogen. Microbial growth in experiments was assayed with alamar blue kits according Immunoblotting (Western Blotting) to the manufacturer's instructions. Briefly, resaZurin reagent 0.138 Samples were first resolved by electrophoresis on was added to microbial cultures (1:10) and samples incubated SDS-PAGE (4-12% Bis-Tris gels) and then transferred to for 30 or 60 minutes. Fluorescence signal was measured at polyvinylidene fluoride membrane. Membranes were excitation of 530 nm and emission at 590 nm. Signal was blocked with bovine serum albumin (10%) then probed with blanked on sterile media. For experiments with brain homo mAb 4G8 (1:200), mAb 6E10 (1:2,000), or mAb anti-LL-37 genates, blank wells contained all components as tests but (1:200) (Hycult Biotechnology, Uden, The Netherlands). Fol were not inoculated with C. albicans. lowing washing, membranes were incubated with goat anti Anti-AB Immunostaining of Bacteria mouse IgG-coupled to HRP. Blots were developed with chemiluminescence reagent (Pierce, Rockford 111.) and sig 0134 E. faecalis smears were air-dried on Superfrost/Plus nal captured using a VersDoc digital imaging system (Bio microscope slides (Fisher Scientific, Pittsburgh, Pa.) and then heated to kill and fix bacterial cells. Fixed cells were incu Rad, Hercules, Calif.). Blot incubations used Tris buffered bated with 3% methanolic hydrogen peroxide for 30 minutes saline, pH 8 containing 0.1% Tween (TBST). at room temperature to inhibit endogenous peroxidase activ Statistical Analysis ity, passed through graded alcohol, and rinsed three times in deionized water and phosphate-buffered saline (PBS). Slides 0.139 Association coefficients between AB levels in brain were then incubated with a 1:2,000 dilution of the anti-AB homogenate and C. albicans growth were calculated using monoclonal antibody (mAb) 6E10 (Covance, Princeton, N.J.) Pearson correlation test and linear regression. Experimental in TBST. Following washing, slides were incubated with goat groups were compared by unpaired two-tailed t-test with a anti-mouse IgG-coupled to HRP (1:200). Detection and nominal alpha criterion level of 0.05. Antimicrobial signal in localization steps were performed using Vectastain ABC kit AD and non-AD cohorts passed a D'Agostino-Pearson test and DAB Substrate Kit (Vector Laboratories, Burlingame, for normality (alpha=0.05) with p values of 0.077 and 0.24, Calif.). respectively. Variances of signal from AD and non-AD cohorts were not significantly different (p=0.18). Alternative Assaying Antimicrobial Activity in Brain Homogenate non-parametric statistical analysis of antimicrobial activity in 0135 Samples of AD (n=32) or non-demented control temporal lobe homogenates by two-tailed Mann-Whitney U (n=13) temporal lobe and cerebellum were homogenized in test also returned a significant difference between AD and US 2012/0058935 A1 Mar. 8, 2012

non-AD cohorts (p=0.018). Statistical analysis used Graph CRAMP in the blood-brain barrier and meninges after Pad Prism software package (La Jolla, Calif.). meningococcal infection. Infect Immun 74: 6982-6991. 0154) 12. Ong PY, Ohtake T. Brandt C, Strickland I, Bogu Other Embodiments niewicz, M. et al. (2002) Endogenous antimicrobial pep tides and skin infections in atopic dermatitis. NEngl J Med 0140 From the foregoing description, it will be apparent 347: 1151-1160. that variations and modifications may be made to the inven (O155 13. Von Haussen J. Koczulla R, Shaykhiev R, Herr tion described herein to adopt it to various usages and condi C, Pinkenburg O, et al. (2008) The host defence peptide tions. Such embodiments are also within the scope of the LL-37/hCAB-18 is a growth factor for lung cancer cells. following claims. Lung Cancer 59:12-23. 0141. The recitation of a listing of elements in any defini 0156 14. Ciornei CD, Tapper H, Bjartell A, Sternby NH, tion of a variable herein includes definitions of that variable as Bodelsson M (2006) Human antimicrobial peptide LL-37 any single element or combination (or Subcombination) of is present in atherosclerotic plaques and induces death of listed elements. The recitation of an embodiment herein vascular smooth muscle cells: a laboratory study. BMC includes that embodiment as any single embodiment or in Cardiovasc Disord 6: 49. combination with any other embodiments orportions thereof. (O157 15. Kirkitadze M D, Bitan G, Teplow D B (2002) 0142. All patents and publications mentioned in this speci Paradigm shifts in Alzheimer's disease and other neurode fication are herein incorporated by reference to the same generative disorders: the emerging role of oligomeric extent as if each independent patent and publication was assemblies. J Neurosci Res 69: 567-577. specifically and individually indicated to be incorporated by 0158 16. Wogulis M. 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E: high-avidity binding to B-amyloid and increased fre NCCLS Document M26-A. Wayne, Pa. quency of type 4 allele in late-onset familial Alzheimer 0226 84. Shiloh MU, Ruan J, Nathan C (1997) Evalua disease. Proc Natl Acad Sci USA 90: 1977-1981. tion of bacterial Survival and phagocyte function with a 0211 69. Urosevic N, Martins R N (2008) Infection and fluorescence-based microplate assay. Infect Immun 65: Alzheimer's disease: the APOE epsilon4 connection and 3.193-31.98. lipid metabolism. JAlzheimers Dis 13: 421-435. 0227 85. Hamid R, Rotshteyn Y. Rabadi L, Parikh R, 0212 70. Bertram L. Lange C. Mullin K. Parkinson M, Bullock P (2004) Comparison of alamar blue and MTT Hsiao M. et al. (2008) Genome-wide association analysis assays for high through-put screening. Toxicol InVitro 18: reveals putative Alzheimer's disease susceptibility loci in 703-710. addition to APOE. Am J Hum Genet. 83: 623-632. 1. A method for treating or preventing a microbial infection 0213) 71. Roberts G. W. Gentleman SM, LynchA, Graham in a subject in need thereof, the method comprising contact DI (1991) 13 A4 amyloid protein deposition in brain after ing the Subject with an effective amount of a B-amyloid pep head trauma. Lancet 338: 1422-1423. tide, oligomer, derivative or analog thereof, thereby prevent 0214. 72. Tesco G. KohY H. Kang E L., Cameron AN, Das ing or inhibiting a microbial infection in the Subject. S, et al. (2007) Depletion of GGA3 stabilizes BACE and 2. The method of claim 1, wherein the microbial infection enhances B-secretase activity. Neuron 54: 721-737. comprises a skin infection and the effective amount is an 0215. 73. Xie Z, DongY. Maeda U, Moir R, Inouye SK, et amount sufficient to treat the subject. al. (2006) Isoflurane-induced apoptosis: a potential patho 3. A method for preventing or inhibiting microbial growth genic link between delirium and dementia. J Gerontol A on a Surface, the method comprising contacting the Surface Biol Sci MedSci 61: 1300-1306. with an effective amount of a 3-amyloid peptide, oligomer, 0216) 74. Kobayashi N. Masuda J. Kudoh J, Shimizu N. derivative or analog thereof, thereby preventing or inhibiting Yoshida T (2008) Binding sites on tau proteins as compo microbial growth on the Surface. nents for antimicrobial peptides. Biocontrol Sci 13:49-56. 4. The method of claim 3, wherein the method cleans, 0217 75. Bolintineanu D. Hazrati E. Davis HT, Lehrer R sanitizes, or disinfects the Surface. I, Kaznessis YN (2009) Antimicrobial mechanism of pore 5. The method of claim 3, wherein the surface is a wall, forming protegrin peptides: 100 pores to kill E. coli. Pep floor, ceiling, counter, machine, or other Surface or equipment tides 31: 1-8. present in an industrial, commercial, or clinical setting. 0218 76. Goumon Y. Lugardon K, Kieffer B. Lefevre J, 6. The method of claim 3, wherein the clinical setting is a Van Dorsselaer A, et al. (1998) Characterization of Anti hospital, medical office, clinic, health center, or laboratory. bacterial COOH-terminal Proenkephalin-A-derived Pep 7. The method of claim3, wherein the surface contacted is tides (PEAP) in Infectious Fluids. Importance of enkelytin, present in a dwelling, School, office building, plane, train, the antibacterial PEAP209-237 secreted by stimulated automobile, restaurant, cafeteria, or daycare center. chromaffin cells. J Biol Chem 273: 2.9847. 8. The method of claim 3, wherein the preventing or inhib 0219 77. Steffen H. Rieg S, Wiedemann I, Kalbacher H, iting microbial growth comprises sanitizing or disinfecting Deeg M. et al. (2006) Naturally processed dermcidin-de and the Surface comprises a body part, the method comprising US 2012/0058935 A1 Mar. 8, 2012

contacting the body part with an effective amount of a B-amy 13. The method of claim 1, wherein the effective amount loid peptide, oligomer, derivative or analog thereof, thereby comprises 1% to 90% of a B-amyloid peptide, oligomer, sanitizing or disinfecting the body part. derivative or analog thereof, or combinations thereof. 9. The method of claim 8, wherein the B-amyloid peptide, 14. An antimicrobial composition comprising an effective oligomer, derivative or analog thereof, is formulated as a skin cleanser. amount of a 3-amyloid peptide, oligomer, derivative orana 10. The method of claim 8, wherein the skin is cleansed in log thereof, thereof in a carrier or diluent. preparation for Surgery. 15. The composition of claim 14, wherein the B-amyloid 11. The method of claim 1, wherein the microbe is a bac peptide, oligomer, derivative or analog thereof, is formulated teria, virus, or fungus. as a water soluble or water insoluble solid, liquid, emulsion, 12. The method of claim 1, wherein the microbe is selected slurry, or powder. from the group consisting of Candida albicans, Escherichia 16. The composition of claim 14, wherein the composition coli, Staphylococcus epidermidis, Streptococcus pneumo is topically administered. niae, Staphylococcus aureus, Listeria monocytogenes, 17. A pharmaceutical pack comprising the composition of Enterococcus faecalis, Streptococcus agalactiae Pseudomo claim 14 in an individual dosage amount. nas aeruginosa, Streptococcus pyogenes, Streptococcus mitis, and Streptococcus salivariu. c c c c c