JP2018008971A - Bismuth-thiol as disinfectant for used in agricultural, industrial and other use - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a bismuth-thiol (BT) compound for protecting a plant against bacteria, fungi or virus pathogen.SOLUTION: There is provided a method for bringing a plant into contact with an effective dose of a bismuth-thiol (BT) compound with sufficient condition and time for protecting injection of the plant by bacteria, fungi or virus pathogen or inhibition of a biofilm by bacteria, fungi or virus pathogen, where the BT composition substantially contains a monodisperse suspension of fine particles containing a BT compound, and the fine particles have a volume average diameter of approximately 0.4-approximately 10 μm; and preferably, a method including a step of bringing the BT composition into contact with the plant, where the step includes bringing synergical or reinforcing antibiotics into contact with the plant simultaneously or successively in an arbitrary order.SELECTED DRAWING: None

Description

CROSS REFERENCE TO RELATED APPLICATIONS This application takes advantage of PCT application PCT / US2011 / 023549, filed February 3, 2010, and US provisional application 61 / 373,188, filed August 12, 2010. Each of which is incorporated herein by reference in its entirety.

  Embodiments of the invention disclosed herein relate to compositions and methods for the treatment of microbial infections. In particular, embodiments of the present invention include improved treatments for managing bacterial infections in agriculture, industry, manufacturing, clinical, personal health care and other aspects, including treatment of bacterial biofilms and other conditions. About.

Description of Related Art A complex series of coordinated cellular and molecular interactions that contribute to the response and resistance to microbial infection and / or to the healing or retention of plant and animal (including human) body tissues are generally External factors such as opportunistic and nosocomial infections (eg, clinical regimens that may increase the risk of infection), local or systemic administration of antibiotics (which may affect cell growth, migration or other functions, antibiotics May be selected for resistant microorganisms) and / or may be adversely affected by other factors.

  Unfortunately, antibiotics introduced systemically or locally are often not effective for the treatment of many chronic bacterial infections and are generally not used unless there is an acute bacterial infection. Current approaches include the administration or application of antibiotics, but such remedies may facilitate the emergence of antibiotic resistant strains and / or may be ineffective for bacterial biofilms . Therefore, it may be particularly important to use disinfectants when drug resistant bacteria (eg, methicillin resistant S. aureus or MRSA) are detected. Although there are many widely used antiseptics, established bacterial populations or subpopulations may not respond to these agents, or any other currently available treatment. In addition, multiple disinfectants may be toxic to host cells at concentrations required to be effective against established bacterial infections, and as such, such disinfectants are inappropriate. This problem can be attributed to natural surfaces such as the surface attributes of commercial and / or agriculturally important plants such as numerous crop plants, and / or body epithelial surfaces such as respiratory tracts such as the respiratory tract, nasopharynx and Attempts to cleanse infections from the laryngeal canal, trachea, lungs, bronchi, bronchioles, alveoli, etc.) or the gastrointestinal tract (eg, oral cavity, esophagus, stomach, intestine, rectum, anus, etc.) or other epithelial surfaces This can be especially serious.

  Of particular concern are infectious organisms composed of bacterial biofilms, a relatively recently recognized bacterial tissue, which is caused by suspension unicellular (plankton) bacteria with significantly different behavioral patterns and gene expression due to cell-cell adhesion. , And organized multicellular communities (biofilms) that are sensitive to environmental drugs such as antibiotics. Biofilms may deploy a biological defense mechanism not found in planktonic bacteria, which can protect the biofilm community from antibiotic and host immune responses. The established biofilm can stop the tissue healing process.

  Common microbiological contaminants lurking in persistent and potentially toxic infections include Staphylococcus aureus including MRSA (methicillin-resistant Staphylococcus aureus), Enterococcus spp., Escherichia coli, Pseudomonas aeruginosa, Streptococcus Genus and Acinetobacter baumannii are included. Some of these organisms show the ability to survive on non-nutritive clinical surfaces for months. S. aureus has been shown to be viable for 4 weeks on dry glass and 3-6 months on dry blood and cotton fibers (Domenico et al., 1999 Infect. Immuno. 67: 664-). 669). Both E. coli and Pseudomonas aeruginosa have been shown to survive longer than S. aureus on dry blood and cotton fibers (previous work).

  Microbial biofilms are associated with a substantial increase in resistance to both fungicides and antibiotics. Biofilm morphology occurs when bacteria and / or fungi attach to the surface. This attachment causes changes in gene transcription, resulting in secretions that are remarkably active and difficult to penetrate the polysaccharide matrix and protect the microorganisms. In addition to having very substantial resistance to antibiotics, biofilms are very resistant to the mammal's immune system. Since biofilms become very difficult to eradicate once established, preventing the formation of biofilms is a very important clinical priority. Recent studies have shown that open wounds can be rapidly contaminated by biofilm. These microbial biofilms are thought to delay wound healing and are likely to be associated with the establishment of serious wound infections.

  Intact and functioning skin and other epithelial tissues (e.g. the generally avascular epithelial surface that forms a barrier between the organism and its external environment, e.g. found in the skin, respiratory and gastrointestinal tract, Maintenance of those found in the inner layers such as glandular tissues is critical to the health and survival of humans and other animals.

Disinfectants based on bismuth thiol (BT) A number of natural products (eg, antibiotics) and synthetic chemicals with antimicrobial properties, particularly antibacterial properties, are known in the art and have chemical structures and Antimicrobial effects, such as the ability to kill microorganisms ("killing" effects such as bactericidal), the ability to stop or damage the growth of microorganisms ("static" effects such as bacteriostatic), or microbial function, eg at the site It is characterized at least in part by its ability to interfere with colonization or infection, exopolysaccharide secretion by bacteria, and / or the transformation from a planktonic population to a biofilm population or the expansion of biofilm formation. Antibiotics, bactericides, disinfectants, etc. (including bismuth-thiol), including factors that affect the selection and use of such compositions, such as bactericidal or bacteriostatic capacity, effective concentration, and risk of toxicity to host tissues Or a BT compound) includes, for example, U.S. Pat. S. Discussed in US Pat. No. 6,582,719.

  Bismuth is a Group V metal and is an antimicrobial element (similar to silver). Bismuth alone may not be therapeutically useful and may exhibit certain inappropriate properties, so it may instead be administered by delivery with complexing agents, carriers, and / or other vehicles, The most common example is Pepto Bismol® in which bismuth is combined (chelated) with hyposalicylate. Combining certain thiol (-SH, sulfhydryl) -containing compounds, such as ethanedithiol and bismuth, to provide exemplary bismuth thiol (BT) compounds, compared to other currently available bismuth preparations, It has been determined in previous studies that antimicrobial activity is improved. There are many thiol compounds that can be used to produce BT (eg, Domenico et al., 2001 Antimicrob. Agent. Chemotherap. 45 (5): 1417-1421, Domenico et al., 1997 Antimicrob. 41 (8): 1697-1703, and U.S. RE 37,793, U.S. 6,248,371, 6,086,921, and 6,380,248. And see, for example, US 6,582,719), several of these preparations can inhibit biofilm formation.

  BT compounds are MRSA (methicillin-resistant Staphylococcus aureus), MRSE (methicillin-resistant S. epidermidis), Mycobacterium tuberculosis, Mycobacterium abium, drug-resistant Pseudomonas aeruginosa, enterotoxigenic Escherichia coli, intestinal hemorrhage Escherichia coli, Klebsiella pneumoniae, Clostridium difficile, Helicobacter pylori, Legionella pneumophila, Enterococcus faecalis, Enterobacter cloacae, Salmonella typhimurium, Proteus bulgaris, Yersinia enterocolitica, Vibrio cholera, And activity against Shigella flexneri has been demonstrated (Domenico et al., 1997 Antimicrob. Agents Chemother. 41: 1 97-1703). There is also evidence of activity against cytomegalovirus, herpes simplex virus type 1 (HSV-1) and HSV-2, and yeast and fungi such as Candida albicans. The role of BT is to reduce bacterial virulence, inhibit or kill broad-spectrum antibiotic-resistant microorganisms (Gram-positive and Gram-negative), prevent biofilm formation, and prevent septic shock Have also been demonstrated in treating sepsis and increasing bacterial susceptibility to previously resistant antibiotics (eg, Domenico et al., 2001 Agent. Chemother. 45: 1417-1421; Domenico et al. , 2000 Infect., Med. 17: 123-127 ;, Antimicrob.Agents & Chemother. Microb. Agent. Chemother. 41 (8): 1697-1703; Domenico et al., 1999 Infect.Immun.67: 664-669; Huang et al.1999 J Antimicrob. , 2003 J Antimicrob. Chemother. 52: 915-919; Wu et al., 2002 Am J Respir Cell Mol Biol.

Despite the availability of BT compounds for well over a period of 10 years, the effective selection of appropriate BT compounds for indications of specific infectious diseases remains an elusive goal, and is specific to specific microorganisms. The behavior of BT cannot be predicted, the synergistic activity of a specific BT and a specific antibiotic against a specific microorganism cannot be predicted, and the in vitro BT effect can usually be predicted from the in vitro BT effect. First, the BT effect on a planktonic (single cell) microbial population cannot be a predictor of the BT effect on a microbial community, for example, a bacterium organized into a biofilm. In addition, limitations such as solubility, tissue permeability, bioavailability, and biodistribution may interfere with the ability to deliver clinical benefits safely and effectively in the case of some BT compounds. There is. The embodiments of the invention disclosed herein address these needs and present other related benefits.
Plant and agricultural product defense: a description of the relevant technical fields

  In the field of agriculture and botany, for biofilms and formulations for reducing disease in plants, and for example in seeds, plants, fruits and flowers, in soil and in cut flowers, trees, fruits, leaves, stems and others There is a recognized need for methods of using such formulations in plant parts.

  In agriculture, billions of dollars of crops are lost each year due to biofilm formation. The problem of anthrax and biofilm-related diseases in plants is well understood, even though many approaches that have attempted to address it do not meet the requirements. Plant diseases are involved in the transport and storage of fruits, vegetables, cut flowers and trees, and other plant products, because the normal defense mechanisms used by intact living plants are no longer effective in harvested products It will also affect the industry.

  Therefore, for agricultural purposes, reducing the amount of microbial growth on the surface of leaves, stems, fruits and flowers in situ, in transportation or in commercial terms, while maintaining compliance with environmental regulations Is desirable. At the same time, in order to enhance the desirable attributes of these products, it is desirable to allow the flow of water within the cut flowers, plants and trees to retain the swelling, integrity and quality of the plant tissue.

  Organisms that cause infectious diseases in plants include fungi, bacteria, viruses, protozoa, nematodes and parasitic plants. Insects and other pests also affect plant health by consumption of plant tissue and by exposure of plant tissue to microorganisms.

  Biofilms typically occur when bacteria bind to a surface in an aqueous environment, such as under water conditions or in water droplets or other high humidity conditions, and after binding, the biofilm formation becomes sticky. It begins to secrete sex substances, which can then be combined with various materials such as metals, plastics, medical implants and tissues. These biofilms can cause a number of problems, including material degradation and pipe clogging in the box office and agricultural environments, and infection of surrounding tissues if it occurs in a medical environment. The medical field is particularly susceptible to problems caused by biofilm formation; implanted medical devices, catheters (urinary, venous, dialysis, heart) and slow healing wounds are more easily caused by bacteria present in the biofilm Infiltrated. In agriculture, biofilms can cause mastitis, Pierce's disease, potato ring rot, various crop downy mildews, and anthracnose in many types of plants. Biofilms also reduce cut flower and tree quality and product life.

  Many plant diseases are caused by soil-specific biofilm-producing bacteria. Most microorganisms in the natural environment are present in multicellular aggregates commonly described as biofilms. Cells adhere to the surface and to each other through a complex matrix comprising various extracellular polymeric substances (EPS), such as exopolysaccharides, proteins and DNA. Plant-associated bacteria interact with host tissue surfaces during pathogenesis and symbiosis, and in a commensal relationship. The observation of bacteria associated with plants incrementally reveals biofilm-type structures that change from a small population of cells to a broad biofilm. Plant tissue surface properties, nutrient and water availability, and the sexual characteristics of cloned bacteria strongly influence the resulting biofilm structure (Ramey et al., 2004 Curr Opinion Microbiol. 7: 602-9).

  The terrestrial environment is inhabited by a rich and diverse microbial population that can compete and reform for resource pools. Plants are colonized by bacteria on their leaves, roots, seeds and internal vascular bundles. Each tissue type has unique chemical and physical properties that represent challenges and opportunities for foreign microorganisms. Biofilms are formed at the time of association or at a later stage and have significant potential to direct or regulate plant-microbe interactions. As many microorganisms actively modify the colonized plant environment, additional temporal and spatial complexity arises.

  Surface associated bacteria have a significant impact on agriculture. In developed countries, losses caused by plant diseases have reached up to 25% of crop yields, a percentage that is much higher than in developing countries. The epidemic population constitutes a reservoir of infection and a future source of infection and can be found on host and non-host plants. Grape vine bacterial pathogens, Xylophilus ampelinus, form a thick biofilm in the vascular bundles of these plants (Grail & Manceau 2003). Xylella fastidiosa is the causative agent of vine pierce disease. Pierce fungus can form biofilms in the xylem tract of many economically important crops. The pathogenic mechanism is largely due to clumping of the xylem canal due to aggregation of pierce fungus and biofilm formation. Vascular blockade is thought to be greatly involved in disease development, and xylem sap provides a natural medium that promotes the virulence of vine piercing and citrus spotted whitening (Zaini et al., 2009 FEMS Microbiol LETT 295: 129-34).

  One of the most relevant plant pathogens, Pseudomonas syringae, causes brown spot disease in beans. It colonizes the leaf surface sporadically in a single subgroup (less than 10 cells), whereas the larger population (greater than 1000 cells) mainly has ciliary processes with higher nutrient availability. Or it develops near the veins. Large aggregates survive drought stress better than single cells. If it does not cause infection in the host plant tissue, Pseudomonas syringae remains alive as a plant epiphyte (ie, a colony of the aerial part of the plant) (Monier et al., PNAS 2003; 100: 15977-82).

  Pseudomonas putida can quickly respond to the presence of root exudates in the soil and assemble at the root colonization site to establish a stable biofilm (Espinosa-Urgel et al., Microbiol). 2002; 148: 341-3).

  The cabbage black rot fungus (Xanthomonas campestris pv. Campestris (Xcc)) produces black roots in cruciform plants and approaches the vascular bundle through the root wound site. Disease toxicity involves degradable exoenzymes and exopolysaccharide xanthan gum necessary for disease toxicity (Dow et al., PNAS 2003; 100: 10995-10000).

  Xanthomonas smitii subsp. Citri is the cause of citrus ulcer disease. This disease has been found on most continents around the world except Europe. The pathogen has been eradicated in many countries. Xanthomonas smithii forms ulcer lesions in the fruits, leaves and twigs of citrus plants. The wind and rain can spread the bacteria to 15 km, open the stomata or wounds and infect citrus trees (Sosnowski, et al., Plant Pathol 2009; 58: 621-35).

  Pantoea stewartii subsp. Stewartii causes Stewart blight in corn and is transmitted by corn flea beetles. Bacteria mainly live in the host xylem and produce large amounts of exopolysaccharides (von Bodman et al., PNAS 1998; 95: 7687-92).

  Ralstonia solanacerum is a soil-borne pathogen that causes fatal killing in many plants. Disease toxicity depends on EPS and cell wall degrading enzymes controlled by a complex regulatory network (Kang et al., Mol Microbiol 2002; 46: 427-37).

  Clavibacter michiganensis subsp. Sepedonicus is a Gram-positive plant pathogen that causes bacterial ring rot of potato. Marques et al. Showed an assembly wrapped in a macrobacterial matrix associated with the xylem canal (Marques et al., Phytopathol 2003; 93: S57).

  The biofilm-producing Erwinia chrysanthemi causes soft rot due to rapid erosion of plant tissue. The production of pectin enzymes is regulated by quorum sensing (QS) and therefore pectin degrading enzyme secretion may be impossible if bacterial aggregates cannot be formed. The related plant pathogen Erwinia amylovara infects about 75 different species of plants, all of the family Rosaceae. Hosts for this bacterium include apples, pears, blackberries, cotoneasters, wild apples, firethorn (Pyracantha), hawthorns, bokeh, rowan, pears, quince, raspberries, zifriboc and snow willow. The cultivated species apple, pear and quince are the most severely affected species. One burn disease outbreak in Michigan in 2000 caused more than 220,000 trees to be lost, for a total loss of $ 42 million. Annual losses and administrative costs for burn disease in the United States are estimated to be over $ 100 million (Norelli et al., Plant Dis 2003; 87: 26-32).

  Erwinia amylobora produces two exopolysaccharides, amyloborane and levan, that cause characteristic burn-off symptoms in the host plant (Koczan et al., Phytopathol 2009; 99: 1237-44). In addition, other genes, and their encoded proteins, have been characterized as virulence factors for Erwinia amylobora that encode enzymes that promote sorbitol metabolism, proteolytic activity and iron acquisition (Oh & Beer. FEMS Microbiology). Lett. 2005; 253: 185-192).

  When a part of a plant is attacked by a microbial phytopathogen, such as a biofilm former, its action is usually to weaken or kill the plant. By infecting leaves, pathogens compromise the plant's ability to produce its food (eg, by photosynthesis). Some plant pathogens block fluid transport ducts in the stalks that feed the leaves, and if such pathogens attack the roots, water and nutrient uptake is reduced or completely stopped The Plant vascular blockage is often accompanied by biofilm-producing bacteria that clog water and nutrient flow, both in plants growing in soil and in plants cut in vase water.

  When a plant is attacked by one of these microorganisms, the resulting damage provides an opportunity for additional microbial invasion of the plant tissue, which is combined to ultimately damage and destroy the plant It is an onslaught. Plants under environmental stress, such as drought or undernutrition, are particularly susceptible to microbial attack.

  Sometimes microbial “infections” are symbiotic, in which case both organisms benefit. A good example of this is the well-known nitrogen-fixing bacterium (rhizobium), which inhabits the root nodules of legumes (peas), which provide nutrients and protection, Bacteria obtain nitrogen from the air and convert it into a form that the host can use. As another example, mycorrhiza is one large eye of a fungus that has a symbiotic relationship with plant roots. In view of such mutually beneficial symbiosis, plant protection or protection against harmful microbial pathogens desirably uses antimicrobial agents that do not disrupt these symbiotic relationships whenever possible.

  The humic fungi are essential for decomposing dead biomaterials to produce the humic substances required for good soil structure. They do not have any chlorophyll and therefore cannot use light to capture energy (eg by photosynthesis); instead, they are plant and animal material (living or dead) The energy is obtained by decomposing. Furthermore, although they can survive in symbiotic relationship with certain plant species, such as the mycorrhiza of the thin roots of cones, they cannot survive without them to take up the nutrients necessary for life support. The widespread use of chemicals to control harmful plant pathogens damages the balance of these beneficial fungi and violates the principles of organic matter management.

  However, there are other less welcome fungi that attack live plants and cause them to be debilitated or killed. Viruses, another class of microbial phytopathogens, inhabit cells of plant tissue and are therefore often unable to be treated with topically applied chemicals, and diseased plants are subject to destruction. There are generally no antibiotics specifically developed for the treatment of plants (though some antibiotics developed for other purposes have found use in plants) and many economically significant Plant species remain vulnerable to attack by pathogenic bacteria. For example, burn disease invasion of many plant species of the Rosaceae proved untreatable. In contrast, a number of harmful fungi can be killed by topically applied chemicals without harming the plant host because of the different fungal growth habits, i.e. a number of undesirable pathogenicities This is because fungi grow on the plant surface using root-like structures to extract nutrients and do not grow in plant tissues.

  Because killing many plant pathogens is often difficult or impossible, many strategies to protect plants against harmful microbial pathogens relieve the philosophy that “prevention is better than cure”. By observing good hygiene when growing and growing plants, many microbial plant diseases can be prevented by blocking the opportunity for microbial infection to be established. Often, significantly lower amounts of insecticide or microbicide can be effective when such agents are used prophylactically rather than in response to established infections.

For example, if the soil is poor in quality (eg, lack of nutrients) by itself or in combination with drought or heavy rain or flood, if the plant is not growing under optimal or near optimal conditions, It becomes easy to get a disease. For example, extreme humid conditions can promote pathogenic fungal and / or bacterial growth. For example, quorum sensing in Pseudomonas syringae is indicated by the availability of water on the leaf surface (Dulla & Lindow. PNAS 2008; 105: 3-082-7). Of course, not all plant diseases are transmitted by insects, others are airborne. For example, aphids and other sucking insects are the primary vectors of viruses. Fungal disease spores are carried in the air and in raindrops and splashes.
Biofilms on seeds and shoots

Bacterial adhesion to seeds is a process that strongly affects rhizosphere colonization. Seed suppliers often deliberately coat seed stocks with microbial biofilms and inoculate the developing rhizosphere. Conversely, biofilms on seeds and shoots used for human consumption are often common sources of gastrointestinal infections. Pseudomonas putida effectively adheres to the seed and then colonizes the rhizosphere. An endoparasite population of non-pathogenic actinomycetes found in wheat tissue was obtained from internal colonization by surface sterilized seed actinomycetes. Other studies of seed colony formation have reported rod-shaped and cocci-type bacteria embedded in EPS in scanning electron micrographs of alfalfa seeds and shoots. Biofilms are resistant to washing in seeds and shoots and other common antibacterial treatments, as is well known. It has been shown that both E. coli O157: H7 and Salmonella populations on alfalfa sprouts require much more severe processing than simply washing to reduce the number of adherent microorganisms, and complete removal is not achieved. Etc. found. Viable bacteria appeared to be inhabited within the biofilm (Ramey et al. Curr Opinion Microbiol 2004; 7: 602-9).
Cut flowers and cut trees

  Vascular pathogens inhabit xylems or phloems of plant hosts and generally depend on insect vectors or wounds for transmission. Cut flowers or cut trees are similar types of wounds that are particularly prone to vascular infection. Biofilm bacteria enter and clog the cut vascular bundles, impeding the flow of water, minerals and nutrients. Cut flower preservatives diluted in vase water often contain salicylate or aspirin to reduce biofilm formation (Domenico et al., J Antimicrob Chemo 1991; 28: 801-10; Salo et al. , Infection 1995; 23: 371-7), providing a low pH to prevent bacterial growth and disrupt the biofilm.

  Antibacterial agent in agriculture. Eradication of plant pathogen invasion is very important for the protection of plant industries, managed gardens and the natural environment around the world. The consequences of pathogens becoming endemic are severe and in some cases can affect the country's economy. Current strategies for eradication of pathogens rely on techniques for the treatment, removal and disposal of affected host plants. There are many examples where these techniques have been successful, but there are many cases where this is not the case. Elaboration relies on a solid understanding of the biology and epidemiology of pathogens and their interactions with the host. In examining examples discussing phytopathogens and affected hosts around the world, especially Australian, various techniques such as burning, filling, composting, soil and biofumigation, solarization, steam sterilization and biological mediators Controls have been used (Sosnowski, et al., Plant Pathol 2009; 58: 621-35).

  Antibiotics have also been used since the 1950s to control certain bacterial diseases of valuable fruits, vegetables and ornamental plants. Today, the most commonly used antibiotics in plants are oxytetracycline and streptomycin. In the United States, antibiotics applied to plants account for less than 0.5% of total antibiotic use. Plant pathogen resistance to oxytetracycline is rare, but the emergence of streptomycin-resistant strains of Erwinia amylobora, Pseudomonas species and Xanthomonas campestris has hampered control of several important diseases. Thus, the role of antibiotic use in plants in the antibiotic resistance crisis in human medicine is the subject of discussion (McManus et al., Annu Rev Phytopathol 2002; 40: 443-65).

  The emergence of streptomycin resistance (SmR) plant pathogens complicates the control of plant bacterial diseases. For example, in the United States streptomycin is Xanthomonas campestris pv. Permitted for tomatoes and peppers for the control of vesicatoria, but resistant strains are now widespread and are rarely used for this purpose. Resistance in the burn pathogen Erwinia amylobora has widespread economic and political implications. Other phytopathogenic bacteria for which SmR has been reported include Pectobacterium carotovora, Pseudomonas chicory, Pseudomonas lacrimans, Pseudomonas syringe pv. Paprance, Pseudomonas syringe pv. Syringe and Xanthomonas dieffenbaquier (McManus et al., Annu Rev Phytopathol 2002; 40: 443-65). The emergence of SmR Erwinia amylobora has increased the burn epidemic in the western United States and Michigan.

  Streptomycin and oxytetracycline have been assigned to the least toxic class by the US Environmental Protection Agency (EPA), and no carcinogenic or mutagenic activity has been observed for either antibiotic.

  Antibiotic alternatives are available and at least to some extent practical. Indeed, bacterial disease management in most crop systems is based on the incorporation of host genetic resistance, hygiene facilities (avoidance or elimination of inoculation), and cultivation practices that create an unfavorable environment for disease outbreaks. Plant biocontrol using a variety of bacterial and fungal species is of increasing interest. Rhizobium is considered an efficient microbial competitor in the root zone. A number of different bacterial genus representatives have been introduced into the soil on seeds, roots, tubers or other plant material to improve crop growth. Examples of these bacterial genera include Acinetobacter, Agrobacterium, Arthrobacter, Azospirillum, Bacillus, Bradyrizobium, Francia, Pseudomonas, Rhizobium, Serratia, Thiobacillus and the like. For example, certain types of Bacillus can induce systemic tolerance in many plants (Chudhary & Johri. Microbiol Res 2009; 164: 493-513).

  Application of copper compounds is effective in reducing populations of some bacterial plant pathogens, but some species become resistant to copper (Cooksey Annu Rev Phytopathol 1990; 28: 201-14) ) Most tree-fruit crops are susceptible to copper damage.

  A number of synthetic and natural therapies exist for various plant diseases. Natural remedies include apple cider vinegar for lesions, powdery mildew and red mold disease; sodium bicarbonate spray for anthrax, early tomato blight, leaf blight, powdery mildew, and as a general fungicide; Neem oil; sulfur; garlic; hydrogen peroxide; compost tea. A number of synthetic chemicals are used to prevent or treat plant diseases, resulting in water soluble or water insoluble formulations. Bactericides include phenoxyarsine or phenalsazine, maleimide, isoindole dicarboximide, aryl arylalkanols, 4-thioxopyrimidine derivatives (US Pat. No. 6,384,040), heterocyclic organosilyl compounds and isothiazolinones. Many fungicides are combined with pyrithione derivatives to make synergistic compounds (eg EP 1468607). Certain isothiazole carboxamides can be used for the control of plant pests (eg, US Pat. No. 6,552,056; WO 2001/064644).

  Understand the toxicity issues of powder or crystalline forms of fungicides, US Pat. No. 29,409 dissolved the fungicides in a liquid solvent and added it to the formulation mixture, from which the end use resin composition Teaches that an object can be processed. Dispersions can be used safely where end-use resin compositions are prepared, but inadvertent use or disposal of liquids can still pose environmental and health hazards. Alternatively, the bactericidal agent can also be administered in a water-soluble thermoplastic resin. A disinfectant can be added to the rigid thermoplastic composition to impart biocidal activity to it to inhibit microbial growth on its surface (US Pat. No. 5,229,124). This is a solid melt blended solution consisting essentially of a fungicide dissolved in a carrier resin that is a copolymer of vinyl alcohol and (alkyleneoxy) acrylate. The disinfectant can be a highly toxic chemical, but its low concentration in the end use product and its retention by the resin composition means that the antibacterial agent in the end use product does not harm humans or animals. Guarantee.

  Isothiazolinones, such as N-alkylbenzenesulfonylcarbamoyl-5-chloroisothiazole derivatives, are often used as agricultural fungicides (eg, US Pat. No. 5,045,555). This fungicide is for example in the paper industry, textile industry, to produce coatings and adhesives, in painting, metal processing, resin industry, timber industry, construction industry, agriculture, forestry, fishery industry, food industry and petroleum industry As well as in medicine. It exhibits strong antibacterial activity and can be added to industrial water, circulating water, raw materials or products in appropriate amounts. Furthermore, it can be used to disinfect and sterilize equipment, industrial facilities, livestock sheds or equipment, and seeds, seedlings and raw materials. Other derivatives of isothiazolone are also known (US Pat. No. 3,523,121 and J. Heterocyclic Chem., 8, 587 (1971)). However, all known derivative compounds are highly toxic to animals and fish, which significantly limits their application.

  Sodium bicarbonate has generally also been found to possess fungicidal properties when applied to plants, but typically requires frequent reapplication to achieve efficacy.

  The role of iron in the plant host-parasite relationship has been elucidated in different diseases as well as soft rot and burn disease induced by Erwinia chrysantemi and Erwinia amylobora, respectively (Expert. Annu Rev Phytopathol 1999; 37: 307). -34). Because of its unique position in biological systems, iron controls the activity of phytopathogens. The production of siderophores by pathogens not only represents a powerful strategy for obtaining iron from host tissues, but can also act as a protective agent against iron toxicity. The need for a host to bind and possibly sequester metals during disease outbreaks is another major issue. Antibacterial agents that interfere with bacterial iron uptake and cellular respiration can play an important role in plant disinfection.

  Numerous natural products (eg antibiotics) and synthetic chemicals with antibacterial, bactericidal and especially antibacterial properties are known and at least some have been chemically and biologically characterized. Exemplary characteristics include the ability to kill microorganisms (bactericidal action), the ability to stop or diminish microbial growth (bacteriostatic action), or the bacterial nature of a metabolite that colonizes or infects a site These include the ability to block microbial functions (anti-biofilm action) such as secretion (some of which give off odors) and / or conversion from a planktonic population to a biofilm population or biofilm expansion. Antibiotics, disinfectants, bactericides, etc. (including bismuth-thiol or BT compounds) are factors that affect the selection and use of such compositions, such as bactericide, bacteriostatic or anti-biofilm efficacy, It is discussed in US Pat. No. 6,582,719, including effective concentrations and risk of toxicity to host tissue.

  Bacterial microcolonies that are protected within the biofilm are typically resistant to antibacterial or disinfectants. For example, the antibiotic dose that kills floating bacteria needs to be increased by a factor of 1,500 to kill the biofilm bacteria. At this high concentration, some antimicrobial agents can be toxic. For example, oxidation of brominated and chlorinated compounds becomes very toxic and corrosive.

  Suppression of the flower rot period is important for the management of burn diseases. In order to cause a flower infection, Erwinia amylobora needs to grow on the surface of the stigma during the establishment. Rain is necessary for infection because it dilutes the sugar on the canola to an osmotic potential that is not inhibitory to Erwinia amylobora. Rain is also important as an agent for the redistribution of bacteria from the stigma to the flower cylinder. These observations suggest that the optimal time to use antibiotic spray is during this epidemic as well as after heavy rain (Johnson & Stockwell. Annu Rev Phytopathol 1998; 36: 227-48). .

  Other bacterial epiphytes also colonize the stigmas where they can interact with and suppress the pathogen's epiphytic growth. A commercial bacterial antagonist of Erwinia amylobora (BrightBan, Pseudomonas fluorescens A506) may be included in the antibiotic spray program. Incorporation of bacterial antagonists by chemical methods represses the population of pathogens and, in a synergistic manner, fills the ecological place provided by the stigmas with non-pathogenic competing microorganisms (Johnson & Stockwell. Annu Rev Phytopathol 1998; 36 : 227-48).

  Pyrithione is a conjugate base derived from 2-mercaptopyridine-N-oxide (CAS # 1121-31-9), which is a derivative of pyridine-N-oxide. Its antifungal action lies in its ability to disrupt membrane transport by blocking the proton pump that activates the transport mechanism. Experiments suggest that fungi can inactivate pyrithione at low concentrations (Chandler & Segel. Antimicrob. Agents Chemother 1978; 14: 60-8). Zinc pyrithione is a zinc coordination complex. This colorless solid is used as an antifungal and antibacterial agent. Because of its low water solubility (8 ppm at neutral pH), zinc pyrithione is suitable for use in outdoor dosages, cement and other products that provide protection against powdery mildew and algae. It is an effective algicide. However, it is chemically incompatible with the amount depending on the metal carboxylate curing agent. When used in latex paints containing water containing large amounts of iron, sequestering agents that selectively bind iron ions are required.

  Of particular concern in agriculture are infections consisting of bacterial biofilms, a fairly recently recognized organization of bacteria, whereby free unicellular ("planktonic") bacteria are indirect Upon arrival, they are assembled into organized multicellular communities (biofilms) that have significantly different behavioral patterns, gene expression, and susceptibility to environmental agents including antibiotics. Biofilms develop biological defense mechanisms not found in planktonic bacteria, which can protect the biofilm community against antibiotics and host immune responses. Established biofilms can stop the growth, development or wound healing process in plants.

  Microbial biofilms are associated with a substantial increase in resistance to both disinfectants and antibiotics. Biofilm morphology occurs when bacteria and / or fungi attach to the surface. This attachment induces a transcriptional alteration of the gene, resulting in the secretion of a polysaccharide matrix that is significantly elastic and difficult to penetrate, protecting the microorganism. Biofilms are very resistant to plant immune defense mechanisms in addition to their very substantial resistance to antibiotics. Biofilms, once established, are very difficult to eradicate and thus preventing biofilm formation is a very important agricultural priority. Recent studies have shown that open wounds become immediately contaminated by biofilm. These microbial biofilms are thought to impair growth, fining and / or wound healing and appear to be highly associated with the probability of severe and often refractory infections.

  Clearly, there is a need for improved compositions and methods for treating and preventing microbial infections in and on plants, including microbial infections that occur as biofilms. Certain embodiments described herein address this need and provide other related advantages.

  As disclosed herein, and without being bound by theory, according to certain embodiments described herein for the first time, bismuth-thiol (BT) compounds are widely used in a wide variety of agriculture. Can be used as a disinfectant for use in industrial, manufacturing and other situations, and in the treatment or prevention of infectious diseases and related symptoms, and in personal health care for personal health care, while at least by BT It also reduces the cost of treating such infections, including savings performed in part by mediated prevention or prevention.

  In certain embodiments also described herein, a bacterial biofilm or a bacterium associated with biofilm formation comprising one or more BT compounds and one or more antibiotic compounds described herein ( For example, a plant or plant tissue (eg, roots, bulbs, stems, leaves, branches, vines, toothpicks, buds, flowers or parts thereof, sprouts, including bacteria that can form or otherwise promote biofilms) , Fruits, seeds, cocoons, etc.) and animal tissues and / or natural and artificial surfaces are contemplated, and according to non-limiting theory, a BT compound appropriately selected based on the present disclosure ( The combination of the antibiotic (s) with the antibiotic provides synergies previously not expected in the antimicrobial (including anti-biofilm) effects of such formulations, and / or Prevention of microorganism infection, such as infections, including bacterial biofilm provides unpredicted enhancing effect for the prevention and / or therapeutically effective treatment.

  Also provided herein for use in these and related embodiments is a bismuth-thiol composition beneficially comprising a substantially monodispersed particulate suspension, and methods for their synthesis and use It is.

  Thus, according to certain embodiments of the invention described herein, a method for protecting a plant against bacterial, fungal or viral pathogens comprising: (i) a bacterial, fungal or viral pathogen Prevention of plant infection by, (ii) inhibition of cell viability or cell proliferation of virtually all planktonic cells of bacterial, fungal or viral pathogens, (iii) inhibition of biofilm formation by bacterial, fungal or viral pathogens And (iv) inhibition of biofilm viability or biofilm growth of substantially all biofilm-type cells of a bacterial, fungal or viral pathogen: plant in conditions and time sufficient for one or more of Or a part thereof (eg, root, bulb, stem, leaf, branch, vine, toothpick, bud, flower or part thereof, shoot, fruit, seed, Etc.) in contact with an effective amount of a bismuth-thiol (BT) composition, wherein the BT composition comprises a substantially monodisperse suspension of microparticles comprising a BT compound. A method is provided comprising a suspension, wherein the microparticles have a volume average diameter of about 0.4 μm to about 10 μm. In a further embodiment, the bacterial pathogen comprises Erwinia amylobora cells. In another embodiment, the bacterial pathogen is an Erwinia amylobora, Xanthomonas campestris pathogenic type dieffenbachiae, Pseudomonas syringae; Selected from Bacter mysiganensis subsp. In certain embodiments, the bacterial pathogen exhibits antibiotic resistance. In certain embodiments, the bacterial pathogen exhibits streptomycin resistance. In certain embodiments, the plant is an edible crop plant, and in certain further embodiments, the edible crop plant is a fruit tree. In a further embodiment, the fruit tree is selected from apple, pear, peach, nectarine, plum, apricot tree. In some embodiments, the food crop plant is a Bamboo banana tree. In other embodiments, the food crop plant is a plant selected from tuberous plants, legumes and grain plants. In a further embodiment, the tuberous plant is selected from potatoes and sweet potatoes. In certain embodiments of the above method, the contacting step is performed one or more times. In a further embodiment, at least one of the contacting steps includes one of spraying the plant, immersing the plant, coating the plant, and painting the plant. In certain other additional embodiments, at least one step of contacting is performed at a plant floret, shoot or growth site. In certain embodiments, at least one step of contacting is performed within 24, 48 or 72 hours of the first flowering in the plant.

  In certain embodiments of the above methods, the BT composition comprises BisBAL, BisEDT, Bis-dimercaprol, Bis-DTT, Bis-2-mercaptoethanol, Bis-DTE, Bis-Pyr, Bis-Ery, Bis-Tol, Bis-BDT, Bis-PDT, Bis-Pyr / Bal, Bis-Pyr / BDT, Bis-Pyr / EDT, Bis-Pyr / PDT, Bis-Pyr / Tol, Bis-Pyr / Ery, Bismuth-1-mercapto- One or more BT compounds selected from 2-propanol and Bis-EDT / 2-hydroxy-1-propanethiol. In certain embodiments, the bacterial pathogen exhibits antibiotic resistance.

  In certain further embodiments of the above methods, the methods involve synergistic or potentiating antibiotics in the plants, simultaneously or sequentially, and in any order, in connection with contacting the plant with the BT composition. Includes contact with a substance. In certain embodiments, the synergistic or potentiating antibiotic is an aminoglycoside antibiotic, a carbapenem antibiotic, a cephalosporin antibiotic, a fluoroquinolone antibiotic, a penicillinase resistant penicillin antibiotic, and an aminopenicillin antibiotic Including antibiotics selected from. In certain embodiments, the synergistic or potentiating antibiotic is an aminoglycoside antibiotic selected from amikacin, arbekacin, gentamicin, kanamycin, neomycin, netilmicin, paromomycin, rhodostreptomycin, streptomycin, tobramycin and apramycin.

  According to certain other embodiments, a method for overcoming antibiotic resistance in a plant in which there is an antibiotic resistant bacterial phytopathogen, comprising: (a) one of the following: Contacting the plant with an effective amount of BT composition for conditions and time sufficient for one or more: (i) prevention of infection of the plant by an antibiotic-resistant bacterial pathogen, (ii) the substance of the antibiotic-resistant bacterial pathogen Inhibition of cell viability or cell proliferation of all planktonic cells, (iii) inhibition of biofilm formation by antibiotic resistant bacterial pathogens, and (iv) virtually all biofilm types of antibiotic resistant bacterial pathogens Inhibition of cell biofilm viability or biofilm growth (wherein the BT composition comprises a substantially monodispersed suspension of microparticles comprising a BT compound; The microparticles have a volume average diameter of about 0.5 μm to about 10 μm): (b) in connection with contacting the plant with the BT composition, simultaneously or sequentially and in any order Contacting the plant with a synergistic or potentiating antibiotic is provided.

  In certain embodiments of the above methods, the bismuth-thiol composition comprises a plurality of microparticles comprising a bismuth-thiol (BT) compound, wherein substantially all of the microparticles have a volume average diameter of about 0.4 μm to about 5 μm. And comprising: (a) a bismuth salt containing bismuth at a concentration of at least 50 mM, under conditions and for a time sufficient to obtain a solution substantially free of solid precipitate, and hydrophilic; Admixing an acidic aqueous solution free of polar or organic solubilizers with (ii) a sufficient amount of ethanol to obtain an admixture comprising about 25% ethanol by volume; and (b) a thiol-containing compound. An ethanolic solution containing is added to the mixture of (a) to obtain a reaction solution (in this case, the thiol-containing compound has sufficient conditions for the formation of a precipitate containing fine particles containing a BT compound. Present in the reaction solution in a molar ratio of about 1: 3 to about 3: 1 with respect to bismuth).

  In some embodiments, the bismuth salt is Bi (NO3) 3. In certain embodiments, the acidic aqueous solution comprises at least 5%, 10%, 15%, 20%, 22% or 22.5% by weight bismuth. In certain embodiments, the acidic aqueous solution is at least 0.5 wt%, 1 wt%, 1.5 wt%, 2 wt%, 2.5 wt%, 3 wt%. Contains 3.5 wt%, 4 wt%, 4.5 wt% or 5 wt% nitric acid. In some embodiments, the thiol-containing compound is 1,2-ethanedithiol, 2,3-dimercaptopropanol, pyrithione, dithioerythritol, 3,4-dimercaptotoluene, 2,3-butanedithiol, 1,3-propane. Dithiol, 2-hydroxypropanethiol, 1-mercapto-2-propanol, dithioerythritol, α-lipoic acid, dithiothreitol, methanethiol (CH3SH [m-mercaptan]), ethanethiol (C2H5SH [e-mercaptan]), 1-propanethiol (C3H7SH [n-P mercaptan]), 2-propanethiol (CH3CH (SH) CH3 [2C3 mercaptan]), butanethiol (C4H9SH [n-butyl mercaptan]), tert-butyl mercaptan (C (C 3) 3SH [t-butyl mercaptan]), pentanethiol (C5H11SH [pentyl mercaptan]), coenzyme A, lipoamide, glutathione, cysteine, cystine, 2-mercaptoethanol, dithiothreitol, dithioerythritol, 2-mercaptoindole, trans Glutaminase, (11-mercaptoundecyl) hexa (ethylene glycol), (11-mercaptoundecyl) tetra (ethylene glycol), (11-mercaptoundecyl) tetra (ethylene glycol) functionalized gold nanoparticles, 1,1 ', 4', 1 "-terphenyl-4-thiol, 1,11-undecanedithiol, 1,16-hexadecanedithiol, technical grade 1,2-ethanedithiol, 1,3-propanedithiol, 1,4-benzenedimethanethiol, 1,4-butanedithiol, 1,4-butanedithioldiacetate, 1,5-pentanedithiol, 1,6-hexanedithiol, 1,8-octanedithiol, 1,9 -Nonanedithiol, adamantanethiol, 1-butanethiol, 1-decanethiol, 1-dodecanethiol, 1-heptanethiol, 1-heptanethiolpurum, 1-hexadecanethiol, 1-hexanethiol, 1-mercapto ( Triethylene glycol), 1-mercapto (triethylene glycol) methyl ether functionalized gold nanoparticles, 1-mercapto-2-propanol, 1-nonanethiol, 1-octadecanethiol, 1-octanethiol, 1-pentadecanethiol, 1-pentanethiol, -Propanethiol, 1-tetradecanethiol, 1-tetradecanethiol plum, 1-undecanethiol, 11- (1H-pyrrol-1-yl) undecane-1-thiol, 11-amino-1-undecanethiol hydrochloride, 11- Bromo-1-undecanethiol, 11-mercapto-1-undecanol, 11-mercaptoundecanoic acid, 11-mercaptoundecyl trifluoroacetate, 11-mercaptoundecyl phosphate, 12-mercaptododecanoic acid, 15-mercaptopentadecanoic acid 16-mercaptohexadecanoic acid, 1H, 1H, 2H, 2H-perfluorodecanethiol, 2,2 '-(ethylenedioxy) diethanethiol, 2,3-butanedithiol, 2-butanethiol, 2-ethylhexanethiol , 2-me Ru-1-propanethiol, 2-methyl-2-propanethiol, 2-phenylethanethiol, 3,3,4,4,5,5,6,6,6-nonafluoro-1-hexanethiolpurum, 3- (Dimethoxymethylsilyl) -1-propanethiol, 3-chloro-1-propanethiol, 3-mercapto-1-propanol, 3-mercapto-2-butanol, 3-mercapto-N-nonylpropionamide, 3-mercaptopropion Acid, 3-mercaptopropyl functionalized silica gel, 3-methyl-1-butanethiol, 4,4'-bis (mercaptomethyl) biphenyl, 4,4'-dimercaptostilbene, 4- (6-mercaptohexyloxy) Benzyl alcohol, 4-cyano-1-butanethiol, 4-mercapto-1-butanol, 6- (ferrocenyl) hexanethiol, 6-mercapto-1-hexanol, 6-mercaptohexanoic acid, 8-mercapto-1-octanol, 8-mercaptooctanoic acid, 9-mercapto-1-nonanol, biphenyl-4,4 ′ -Dithiol, 3-mercaptopropionate butyl, copper 1-butanethiolate (I), cyclohexanethiol, cyclopentanethiol, decanethiol functionalized silver nanoparticles, dodecanethiol functionalized gold nanoparticles, dodecanethiol functionalized Silver nanoparticles, hexa (ethylene glycol) mono-11- (acetylthio) undecyl ether, mercaptosuccinic acid, methyl 3-mercaptopropionate, NanoTether BPA-HH, NanoThinks ™ 18, NanoThinks (trade) ) 8, NanoThinks ™ ACID11, NanoThinks ™ ACID16, NanoThinks ™ ALCO11, NanoThinks ™ THIO8, Octanethiol functionalized gold nanoparticles, PEG dithiol average Mn 8,000, PEG dithiol average molecular weight 1,500 PEG dithiol average molecular weight 3,400, S- (11-bromoundecyl) thioacetate, S- (4-cyanobutyl) thioacetate, thiophenol, triethylene glycol mono-11-mercaptoundecyl ether, trimethylolpropane tris (3 -Mercaptopropionate), [11- (methylcarbonylthio) undecyl] tetra (ethylene glycol), m-carborane-9-thiol, p-terfeny And one or more agents selected from ru-4,4 "-dithiol, tert-dodecyl mercaptan, and tert-nonyl mercaptan.

  In certain embodiments, the bacterial pathogen is: (i) one or more gram negative bacteria; (ii) one or more gram positive bacteria; (iii) one or more antibiotic sensitive bacteria; (iv) 1 (V) Staphylococcus aureus, MRSA (methicillin-resistant Staphylococcus aureus), Staphylococcus epidermidis, MRSE (methicillin-resistant Staphylococcus epidermidis), Mycobacterium tuberculosis, Avian tuberculosis, Pseudomonas aeruginosa , Drug-resistant Pseudomonas aeruginosa, Escherichia coli, enterotoxic Escherichia coli, enterohemorrhagic Escherichia coli, Neisseria pneumoniae, Clostridium difficile, Helicobacter pylori, Legionella pneumophila, faecal streptococci, methicillin-sensitive fecal streptococci, Enterobacter cloacae, Salmonella typhimurium Fungus, Proteus vulgaris, Yersinia enterocolitica, Vibrio cholerae, Shigella flexner, Van Mycin-resistant enterococci (VRE), Burkholderia cepacia group, wild gonorrhea, Bacillus anthracis, plague, Pseudomonas aeruginosa, Streptococcus pneumoniae, penicillin-resistant Streptococcus pneumoniae, Escherichia coli, Burkholderia cepacia, Burkholderia -Containing at least one of the following: bacterial pathogens selected from multiboranes, scabs and Acinetobacter baumannii.

  In certain embodiments, the method comprises: (i) a synergistic antibiotic and a plant in combination with the step of contacting the surface and the BT composition, simultaneously or sequentially, and in any order. (Ii) contacting with at least one of the cooperative antimicrobial efficacy enhancing antibiotics. In a further embodiment, the synergistic antibiotic or cooperative antibacterial efficacy enhancing antibiotic is an aminoglycoside antibiotic, a carbapenem antibiotic, a cephalosporin antibiotic, a fluoroquinolone antibiotic, a glycopeptide antibiotic An antibiotic selected from lincosamide antibiotics, penicillinase resistant penicillin antibiotics, and aminopenicillin antibiotics. In a further embodiment, the synergistic antibiotic or the cooperative antibacterial efficacy enhancing antibiotic is an aminoglycoside selected from amikacin, arbekacin, gentamicin, kanamycin, neomycin, netilmycin, paromomycin, rhodostreptomycin, streptomycin, tobramycin and apramycin. Antibiotics.

  In certain other embodiments, a method for overcoming antibiotic resistance in a plant in which there is an antibiotic resistant bacterial pathogen, comprising: (i) prevention of infection of the plant by the bacterial pathogen; (Ii) inhibition of cell viability or cell proliferation of substantially all planktonic cells of bacterial pathogens, (iii) inhibition of biofilm formation by bacterial pathogens, and (iv) substantially all biofilms of bacterial pathogens Inhibiting plant cell biofilm viability or biofilm growth: an effective amount of (1) at least one bismuth-thiol (BT) composition under conditions and time sufficient for one or more of And (2) contacting with at least one antibiotic capable of synergistically enhancing or acting with at least one BT composition. (Wherein the BT composition is a bismuth-thiol composition comprising a plurality of microparticles comprising a bismuth-thiol (BT) compound, wherein substantially all of the microparticles are from about 0.4 μm to about 5 μm. And thereby overcoming antibiotic resistance of the epithelial tissue surface is provided. In a further embodiment, the bacterial pathogen is resistant to an antibiotic selected from methicillin, vancomycin, nafcillin, gentamicin, ampicillin, chloramphenicol, doxycycline, tobramycin, clindamycin and gatifloxacin. In certain other embodiments, the BT composition comprises BisBAL, BisEDT, Bis-dimercaprol, Bis-DTT, Bis-2-mercaptoethanol, Bis-DTE, Bis-Pyr, Bis-Ery, Bis-Tol, Bis. -BDT, Bis-PDT, Bis-Pyr / Bal, Bis-Pyr / BDT, Bis-Pyr / EDT, Bis-Pyr / PDT, Bis-Pyr / Tol, Bis-Pyr / Ery, Bismuth-1-mercapto-2 One or more BT compounds selected from -propanol and Bis-EDT / 2-hydroxy-1-propanethiol are included. In certain embodiments, the synergistic or potentiating antibiotic is clindamycin, gatifloxacin, aminoglycoside antibiotic, carbapenem antibiotic, cephalosporin antibiotic, fluoroquinolone antibiotic, penicillinase resistant penicillin antibiotic Selected from substances and aminopenicillin antibiotics. In a further embodiment, the synergistic or potentiating antibiotic is an aminoglycoside antibiotic selected from amikacin, arbekacin, gentamicin, kanamycin, neomycin, netilmicin, paromomycin, rhodostreptomycin, streptomycin, tobramycin and apramycin. .

  According to certain other embodiments, a bismuth-thiol composition comprising a plurality of microparticles comprising a bismuth-thiol (BT) compound is provided, wherein substantially all of the microparticles have a volume of about 0.4 μm to about 5 μm. Having a mean diameter, the BT compound includes a bismuth or bismuth salt and a thiol-containing compound. In another embodiment, a bismuth-thiol composition comprising a plurality of microparticles comprising a bismuth-thiol (BT) compound is provided, wherein substantially all of the microparticles have a volume average diameter of about 0.4 μm to about 5 μm. Having (a) a bismuth salt containing bismuth at a concentration of at least 50 mM, under conditions and time sufficient to obtain a solution substantially free of solid precipitates, hydrophilic, polar or organic An acidic aqueous solution free of soluble solubilizers to obtain a blend comprising (ii) at least about 5%, 10%, 15%, 20%, 25% or 30% ethanol by volume And (b) a molar ratio of about 1: 3 to about 3: 1 with respect to bismuth under conditions and time sufficient to form a precipitate containing microparticles containing the BT compound. And adding an ethanolic solution containing a thiol-containing compound present in the reaction solution to the mixture of (a) to obtain a reaction solution. In certain embodiments, the bismuth salt is Bi (NO3) 3. In certain embodiments, the acidic aqueous solution comprises at least 5 wt%, 10 wt%, 15 wt%, 20 wt%, 22 wt% or 22.5 wt% bismuth. In certain embodiments, the acidic aqueous solution is at least 0.5 wt%, 1 wt%, 1.5 wt%, 2 wt%, 2.5 wt%, 3 wt%. Contains 3.5 wt%, 4 wt%, 4.5 wt% or 5 wt% nitric acid. In certain embodiments, the thiol-containing compound is 1,2-ethanedithiol, 2,3-dimercaptopropanol, pyrithione, dithioerythritol, 3,4-dimercaptotoluene, 2,3-butanedithiol, 1,3- One or more agents selected from propanedithiol, 2-hydroxypropanethiol, 1-mercapto-2-propanol, α-lipoic acid, and dithiothreitol are included.

  In another embodiment, a method of preparing a bismuth-thiol composition comprising a plurality of microparticles comprising a bismuth-thiol (BT) compound is provided, wherein substantially all of the microparticles are from about 0.4 μm to about 5 μm. A bismuth salt having a volume average diameter, wherein the method comprises (a) conditions and time sufficient to obtain a solution substantially free of solid precipitates, and (i) a bismuth salt having a concentration of at least 50 mM. An acidic aqueous solution comprising no hydrophilic, polar or organic solubilizer, (ii) at least about 5% by volume, 10% by volume, 15% by volume, 20% by volume, 25% by volume or 30% by volume ethanol. Admixing with a sufficient amount of ethanol to obtain an admixture comprising; and (b) bismuth with conditions and time sufficient to form a precipitate comprising microparticles comprising a BT compound. Adding an ethanolic solution containing a thiol-containing compound present in the reaction solution in a molar ratio of about 1: 3 to about 3: 1 to the blend of (a) to obtain a reaction solution. In certain embodiments, the method further includes recovering the precipitate to remove impurities. In certain embodiments, the bismuth salt is Bi (NO3) 3. In certain embodiments, the acidic aqueous solution comprises at least 5 wt%, 10 wt%, 15 wt%, 20 wt%, 22 wt% or 22.5 wt% bismuth. In certain embodiments, the acidic aqueous solution is at least 0.5 wt%, 1 wt%, 1.5 wt%, 2 wt%, 2.5 wt%, 3 wt%. Contains 3.5 wt%, 4 wt%, 4.5 wt% or 5 wt% nitric acid. In certain embodiments, the thiol-containing compound is 1,2-ethanedithiol, 2,3-dimercaptopropanol, pyrithione, dithioerythritol, 3,4-dimercaptotoluene, 2,3-butanedithiol, 1,3- Propanedithiol, 2-hydroxypropanethiol, 1-mercapto-2-propanol, dithiothreitol, α-lipoic acid, methanethiol (CH3SH [m-mercaptan]), ethanethiol (C2H5SH [e-mercaptan]), 1- Propanethiol (C3H7SH [n-P mercaptan]), 2-propanethiol (CH3CH (SH) CH3 [2C3 mercaptan]), butanethiol (C4H9SH [n-butyl mercaptan]), tert-butyl mercaptan (C (CH3) 3SH) [T Butyl mercaptan]), pentanethiol (C5H11SH [pentyl mercaptan]), coenzyme A, lipoamide, glutathione, cysteine, cystine, 2-mercaptoethanol, dithiothreitol, 2-mercaptoindole, transglutaminase, (11-mercaptoundecyl) Hexa (ethylene glycol), (11-mercaptoundecyl) tetra (ethylene glycol), (11-mercaptoundecyl) tetra (ethylene glycol) functionalized gold nanoparticles, 1,1 ′, 4 ′, 1 ″- Terphenyl-4-thiol, 1,11-undecanedithiol, 1,16-hexadecanedithiol, technical grade 1,2-ethanedithiol, 1,3-propanedithiol, 1,4-benzenedimethanethiol, 1 , 4-butanedithiol, 1,4-butanedithiol diacetate, 1,5-pentanedithiol, 1,6-hexanedithiol, 1,8-octanedithiol, 1,9-nonanedithiol, adamantanethiol, 1-butanethiol 1-decanethiol, 1-dodecanethiol, 1-heptanethiol, 1-heptanethiolpurum, 1-hexadecanethiol, 1-hexanethiol, 1-mercapto (triethylene glycol), 1-mercapto (triethylene) Glycol) methyl ether functionalized gold nanoparticles, 1-mercapto-2-propanol, 1-nonanethiol, 1-octadecanethiol, 1-octanethiol, 1-pentadecanethiol, 1-pentanethiol, 1-propanethiol, 1 -Tetradecane All, 1-tetradecanethiolpurum, 1-undecanethiol, 11- (1H-pyrrol-1-yl) undecane-1-thiol, 11-amino-1-undecanethiol hydrochloride, 11-bromo-1-undecanethiol, 11-mercapto-1-undecanol, 11-mercaptoundecanoic acid, 11-mercaptoundecyl trifluoroacetate, 11-mercaptoundecyl phosphate, 12-mercaptododecanoic acid, 15-mercaptopentadecanoic acid, 16-mercaptohexadecanoic acid, 1H, 1H, 2H, 2H-perfluorodecanethiol, 2,2 '-(ethylenedioxy) diethanethiol, 2,3-butanedithiol, 2-butanethiol, 2-ethylhexanethiol, 2-methyl-1- Propanethiol, 2-methyl- -Propanethiol, 2-phenylethanethiol, 3,3,4,4,5,5,6,6,6-nonafluoro-1-hexanethiolpurum, 3- (dimethoxymethylsilyl) -1-propanethiol, 3 -Chloro-1-propanethiol, 3-mercapto-1-propanol, 3-mercapto-2-butanol, 3-mercapto-N-nonylpropionamide, 3-mercaptopropionic acid, 3-mercaptopropyl functionalized silica gel, 3 -Methyl-1-butanethiol, 4,4'-bis (mercaptomethyl) biphenyl, 4,4'-dimercaptostilbene, 4- (6-mercaptohexyloxy) benzyl alcohol, 4-cyano-1-butanethiol, 4-mercapto-1-butanol, 6- (ferrocenyl) hexanethiol, 6 -Mercapto-1-hexanol, 6-mercaptohexanoic acid, 8-mercapto-1-octanol, 8-mercaptooctanoic acid, 9-mercapto-1-nonanol, biphenyl-4,4'-dithiol, butyl 3-mercaptopropionate 1-butanethiolate copper (I), cyclohexanethiol, cyclopentanethiol, decanethiol functionalized silver nanoparticles, dodecanethiol functionalized gold nanoparticles, dodecanethiol functionalized silver nanoparticles, hexa (ethylene glycol) Mono-11- (acetylthio) undecyl ether, mercaptosuccinic acid, methyl 3-mercaptopropionate, NanoTether BPA-HH, NanoThinks (TM) 18, NanoThinks (TM) 8, NanoThinks (TM) AC D11, NanoThinks ™ ACID16, NanoThinks ™ ALCO11, NanoThinks ™ THIO8, octanethiol functionalized gold nanoparticles, PEG dithiol average Mn 8,000, PEG dithiol average molecular weight 1,500, PEG dithiol average molecular weight 3, 400, S- (11-bromoundecyl) thioacetate, S- (4-cyanobutyl) thioacetate, thiophenol, triethylene glycol mono-11-mercaptoundecyl ether, trimethylolpropane tris (3-mercaptopropionate), [11- (Methylcarbonylthio) undecyl] tetra (ethylene glycol), m-carborane-9-thiol, p-terphenyl-4,4 ″ -dithiol, tert-do One or more drugs selected from the group consisting of decyl mercaptan and tert-nonyl mercaptan are included.

  In another embodiment, (i) prevention of surface infection by bacterial, fungal or viral pathogens; (ii) inhibition of cell viability or cell growth of substantially all planktonic cells of bacterial, fungal or viral pathogens; (Iii) inhibition of biofilm formation by bacterial, fungal or viral pathogenesis, and (iv) inhibition of biofilm viability or biofilm development of virtually all biofilm type cells of bacterial, fungal or viral pathogenesis Contacting the epithelial tissue surface with an effective amount of BT composition at conditions and time sufficient for one or more of the BT composition, wherein the BT composition comprises a plurality of microparticles comprising a bismuth-thiol (BT) compound Bacterial, fungal and viral diseases, wherein substantially all of the microparticles have a volume average diameter of about 0.4 μm to about 5 μm For one or more of the species, a natural or artificial surface, such as a biological tissue surface, such as a plant surface (eg, root, bulb, stem, leaf, branch, vine, branch, bud, flower or part thereof, sprout , All or part of the surface of fruits, seeds, pods, etc.) or epithelial tissue surfaces are provided. In certain embodiments, the bacterial pathogen is (i) one or more gram negative bacteria; (ii) one or more gram positive bacteria; (iii) one or more antibiotic sensitive bacteria; (iv) one or more bacteria. (V) Staphylococcus aureus (Staphylococcus aureus), MRSA (methicillin-resistant Staphylococcus aureus), Staphylococcus epidermidis, MRSE (methicillin-resistant S. epidermidis), Mycobacterium tuberculosis Cis, Mycobacterium avium, Pseudomonas aeruginosa, drug-resistant Pseudomonas aeruginosa, Escherichia coli, Enterotoxigenic Escherichia coli, Enterohemorrhagic Escherichia coli, Klebsiella pneumoniae, Clostridium difficile, Helicobacter pylori, Legionella pneumophila, Enterococcus Fecarlis, Mechisiri Sensitive Enterococcus faecalis, Enterobacter cloacae, Salmonella typhimurium, Proteus vulgaris, Yersinia enterocolitica, Vibrio cholere, Shigella flexneri, vancomycin-resistant enterococcus (VRE), Burkholderia cepacia group, Francisella Tularensis, Bacillus anthracis, Yersinia pestis, Pseudomonas aeruginosa, Vancomycin-resistant Enterococcus sp. A small number of bacterial pathogens selected from smegmatis and Acinetobacter baumannii Kutomo including one a. In certain embodiments, the bacterial pathogen exhibits antibiotic resistance. In certain embodiments, the bacterial pathogen is resistant to an antibiotic selected from methicillin, vancomycin, nafcillin, gentamicin, ampicillin, chloramphenicol, doxycycline and tobramycin.

  In certain embodiments, the natural or artificial surface is a mouth / oral surface, artificial device, ceramic, plastic, polymer, rubber, metal product, painted surface, marine structure, such as a hull, rudder, propeller, anchor, hold, Includes ballast tanks, docks, dry docks, wharfs, piles, revetments, or other natural or artificial surfaces.

  In certain embodiments, the surface comprises an epithelial tissue surface comprising tissue selected from the epidermis, dermis, airways, gastrointestinal tract and glandular lining.

  In certain embodiments, the contacting step is performed one or more times. In certain embodiments, the at least one contacting step includes one of spraying, irrigating, dipping and painting a natural or artificial surface. In certain embodiments, the at least one contacting step includes one of inhalation, ingestion, and oral irrigation. In certain embodiments, the at least one contact step comprises topical, intraperitoneal, oral, parenteral, intravenous, intraarterial, transdermal, sublingual, subcutaneous, intramuscular, buccal, intranasal, inhalation, ocular Administration by a route selected from internal, intraauricular, intraventricular, intrafat, intraarticular and intrathecal. In certain embodiments, the BT composition comprises BisBAL, BisEDT, Bis-dimercaprol, Bis-DTT, Bis-2-mercaptoethanol, Bis-DTE, Bis-Pyr, Bis-Ery, Bis-Tol, Bis- BDT, Bis-PDT, Bis-Pyr / Bal, Bis-Pyr / BDT, Bis-Pyr / EDT, Bis-Pyr / PDT, Bis-Pyr / Tol, Bis-Pyr / Ery, Bismuth-1-mercapto-2- One or more BT compounds selected from the group consisting of propanol and Bis-EDT / 2-hydroxy-1-propanethiol are included.

  In certain embodiments, the bacterial pathogen exhibits antibiotic resistance. In other specific embodiments, the method comprises synergizing the natural surface and / or in any order simultaneously or sequentially with respect to contacting the natural or artificial surface with the BT composition. It further comprises contacting with an enhancing antibiotic. In certain embodiments, the synergistic and / or potentiating antibiotics are aminoglycoside antibiotics, carbapenem antibiotics, cephalosporin antibiotics, fluoroquinolone antibiotics, glycopeptide antibiotics, lincosamides An antibiotic selected from antibiotics, penicillinase-resistant penicillin antibiotics, and aminopenicillin antibiotics. In certain embodiments, the synergistic and / or potentiating antibiotic is an aminoglycoside antibiotic selected from amikacin, arbekacin, gentamicin, kanamycin, neomycin, netilmycin, paromomycin, rhodostreptomycin, streptomycin, tobramycin and apramycin It is.

  In another embodiment of the invention described herein, (i) prevention of infection by a bacterial pathogen on a surface on which an antibiotic-resistant bacterial pathogen is present; (ii) substantially all planktonic properties of the bacterial pathogen Inhibition of cell viability or cell growth of cells, (iii) inhibition of biofilm formation by bacterial pathogenesis, and (iv) inhibition of biofilm viability or biofilm growth of virtually all biofilm type cells of bacterial pathogenesis A natural or artificial surface in which an antibiotic-resistant bacterial pathogen is present at a condition and for a time sufficient for one or more of: an effective amount of (1) at least one bismuth-thiol (BT) composition and (2) Simultaneously with at least one antibiotic that is enhanced by and / or can act synergistically with at least one BT composition; Comprises contacting in any order sequentially, wherein the BT composition comprises a plurality of microparticles comprising a bismuth-thiol (BT) compound, wherein substantially all of the microparticles are from about 0.4 μm to about 5 μm. Overcoming antibiotic resistance on the surface of epithelial tissue, overcoming antibiotic resistance on natural or artificial surfaces where antibiotic resistant bacterial pathogens exist (eg, for bacteria of the same species) For bacterial pathogens resistant to at least one antibacterial effect of at least one antibiotic known to have an antibacterial effect against the pathogen) . In certain embodiments, the bacterial pathogen is (i) one or more gram negative bacteria; (ii) one or more gram positive bacteria; (iii) one or more antibiotic sensitive bacteria; (iv) one or more bacteria. (V) Staphylococcus aureus (Staphylococcus aureus), MRSA (methicillin-resistant Staphylococcus aureus), Staphylococcus epidermidis, MRSE (methicillin-resistant S. epidermidis), Mycobacterium tuberculosis Cis, Mycobacterium avium, Pseudomonas aeruginosa, drug-resistant Pseudomonas aeruginosa, Escherichia coli, Enterotoxigenic Escherichia coli, Enterohemorrhagic Escherichia coli, Klebsiella pneumoniae, Clostridium difficile, Helicobacter pylori, Legionella pneumophila, Enterococcus Fecarlis, Mechisiri Sensitive Enterococcus faecalis, Enterobacter cloacae, Salmonella typhimurium, Proteus vulgaris, Yersinia enterocolitica, Vibrio cholere, Shigella flexneri, vancomycin-resistant enterococcus (VRE), Burkholderia cepacia group, Francisella Tularensis, Bacillus anthracis, Yersinia pestis, Pseudomonas aeruginosa, Vancomycin-resistant Enterococcus sp. A small number of bacterial pathogens selected from smegmatis and Acinetobacter baumannii Kutomo including the one.

  In certain embodiments, the bacterial pathogen is resistant to an antibiotic selected from methicillin, vancomycin, nafcillin, gentamicin, ampicillin, chloramphenicol, doxycycline, tobramycin, clindamycin and gatifloxacin.

  In certain embodiments, the natural or artificial surface is an intraoral / oral surface, artificial device, ceramic, plastic, polymer, rubber, metal product, painted surface, marine structure, such as a hull, tire, propeller, anchor, hold, Includes ballast tanks, docks, dry docks, wharfs, piles, revetments, or other natural or artificial surfaces.

  In certain embodiments, the surface comprises a tissue selected from the group consisting of epidermis, dermis, respiratory tract, gastrointestinal tract, and glandular lining. In certain embodiments, the contacting step is performed one or more times. In certain embodiments, the at least one contacting step includes one of spraying, irrigating, dipping and painting the surface. In other specific embodiments, the at least one contacting step includes one of inhalation, ingestion, and oral irrigation. In certain embodiments, the at least one contact step comprises topical, intraperitoneal, oral, parenteral, intravenous, intraarterial, transdermal, sublingual, subcutaneous, intramuscular, buccal, intranasal, inhalation, ocular Administration by a route selected from internal, intraauricular, intraventricular, intrafat, intraarticular and intrathecal. In certain embodiments, the BT composition comprises BisBAL, BisEDT, Bis-dimercaprol, Bis-DTT, Bis-2-mercaptoethanol, Bis-DTE, Bis-Pyr, Bis-Ery, Bis-Tol, Bis- BDT, Bis-PDT, Bis-Pyr / Bal, Bis-Pyr / BDT, Bis-Pyr / EDT, Bis-Pyr / PDT, Bis-Pyr / Tol, Bis-Pyr / Ery, Bismuth-1-mercapto-2- One or more BT compounds selected from propanol and Bis-EDT / 2-hydroxy-1-propanethiol are included. In certain embodiments, the synergistic and / or potentiating antibiotics are clindamycin, gatifloxacin, aminoglycoside antibiotics, carbapenem antibiotics, cephalosporin antibiotics, fluoroquinolone antibiotics, An antibiotic selected from glycopeptide antibiotics, lincosamide antibiotics, penicillinase-resistant penicillin antibiotics, and aminopenicillin antibiotics. In certain embodiments, the synergistic and / or potentiating antibiotic is an aminoglycoside antibiotic selected from amikacin, arbekacin, gentamicin, kanamycin, neomycin, netilmycin, paromomycin, rhodostreptomycin, streptomycin, tobramycin and apramycin It is.

  In another embodiment, (a) at least one BT compound; (b) at least one antibiotic compound that is potentiated and / or capable of acting synergistically with the BT compound; and (C) A disinfecting composition is provided comprising a pharmaceutically acceptable excipient or carrier, including a carrier for topical use. In certain embodiments, the BT compound is BisBAL, BisEDT, Bis-dimercaprol, Bis-DTT, Bis-2-mercaptoethanol, Bis-DTE, Bis-Pyr, Bis-Ery, Bis-Tol, Bis-BDT. Bis-PDT, Bis-Pyr / Bal, Bis-Pyr / BDT, Bis-Pyr / EDT, Bis-Pyr / PDT, Bis-Pyr / Tol, Bis-Pyr / Ery, Bismuth-1-mercapto-2-propanol And Bis-EDT / 2-hydroxy-1-propanethiol. In certain embodiments, the BT composition comprises a plurality of microparticles comprising a bismuth-thiol (BT) compound, and substantially all of the microparticles have a volume average diameter of about 0.4 μm to about 5 μm. In certain embodiments, the BT compound is selected from BisEDT and BisBAL. In certain embodiments, the antibiotic compound comprises an antibiotic selected from methicillin, vancomycin, nafcillin, gentamicin, ampicillin, chloramphenicol, doxycycline, tobramycin, clindamycin, gatifloxacin and aminoglycoside antibiotics. . In certain embodiments, the aminoglycoside antibiotic is selected from amikacin, arbekacin, gentamicin, kanamycin, neomycin, netilmycin, paromomycin, rhodostreptomycin, streptomycin, tobramycin and apramycin. In certain embodiments, the aminoglycoside antibiotic is amikacin.

  In other specific embodiments, (a) bacterial infections on or in the surface are: (i) gram positive bacteria, (ii) gram negative bacteria, and (iii) both (i) and (ii), Natural to support or contain a bacterial biofilm comprising: identifying to include one of: and (b) administering to a surface a formulation comprising one or more bismuth thiol (BT) compositions Or a method of treating an artificial surface, wherein (i) when the bacterial infection comprises gram positive bacteria, the formulation comprises at least one BT compound and at least one antibiotic that is rifamycin. Comprising a therapeutically effective amount, and (ii) if the bacterial infection comprises a gram negative bacterium, the formulation comprises a therapeutically effective amount of at least one BT compound and amikacin, and (iii) a bacterial infection If is including both Gram-positive and Gram-negative bacteria, the formulation BT compounds comprise one or more therapeutically effective amount of a rifamycin and amikacin, thereby treating the surface.

  In certain embodiments, the biofilm comprises one or more antibiotic resistant bacteria. In certain embodiments, treating the surface comprises (i) eradicating bacterial biofilm, (ii) reducing bacterial biofilm, and (iii) inhibiting bacterial biofilm growth. Including at least one of them. In certain embodiments, the BT composition comprises a plurality of microparticles comprising a bismuth-thiol (BT) compound, and substantially all of the microparticles have a volume average diameter of about 0.4 μm to about 5 μm.

  These and other aspects of the embodiments of the invention described herein will become apparent by reference to the following detailed description and accompanying drawings. U. S. RE37,793, U.S. Pat. S. No. 6,248,371, U.S. Pat. S. No. 6,086,921, and U.S. Pat. S. U.S. Patents and U.S. Patent Application Publications, U.S. Patent Applications, Foreign Patents, Foreign Patent Applications, Non-Patent Issuances Referenced herein and / or listed in the Application Data Sheet, including 6,380,248 All of the objects are hereby incorporated by reference in their entirety, so that each is individually incorporated. Aspects and embodiments of the present invention can be modified as needed to provide further embodiments using the concepts of various patents, applications and publications.

Brief description of multiple appearances of drawings
Pseudomonas aeruginosa colony biofilm survival (log CFU; colony forming units) obtained after growth for 24 hours at 37 ° C. on 10% trypsin soy agar (TSA) followed by the indicated treatment for 18 hours Indicates. The indicated antibiotic treatments are: TOB, tobramycin 10 × MIC; AMK, amikacin, 100 × MIC; IPM, imipenem 10 × MIC; CEF, cefepime 10 × MIC; CIP, ciprofloxacin, 100 × MIC; Cpd 2B Compound 2B (Bis-BAL, 1: 1.5). (MIC; minimum inhibitory concentration, eg, lowest concentration to prevent bacterial growth). The number of surviving staphylococcus aureus colony biofilms (logCFU) obtained after growth on 10% trypsin soy agar for 24 hours and subsequent treatment as indicated is shown. Antibiotic treatments indicated were: rifamycin, RIF 100 × MIC; daptomycin, DAP 320 × MIC; minocycline, MIN 100 × MIC; ampicillin, AMP 10 × MIC; vancomycin, VAN 10 × MIC; Cpd 2B, compound 2B ( Bis-BAL, 1: 1.5), Cpd8-2, compound 8-2 (Bis-Pyr / BDT, 1: 1 / 0.5). Figure 2 shows scratch closure over time of keratinocytes exposed to biofilm. (*) Significantly different from control (P <0.001). Figure 2 shows subinhibited BisEDT that suppresses antibiotic resistance to multiple antibiotics. Figure 6 shows the effect of antibiotics with or without BisEDT on the MRSA (methicillin resistant Staphylococcus aureus) flora. Panel A shows only a standard antibiotic immersion disc. [GM = gentamicin, CZ = cefazoline, FEP = cefepime, IPM = imipenem, SAM = ampicillin / sulbactam], LVX = levofloxacin. Figure 2 shows subinhibited BisEDT that suppresses antibiotic resistance to multiple antibiotics. Figure 6 shows the effect of antibiotics with or without BisEDT on the MRSA (methicillin resistant Staphylococcus aureus) flora. Panel B shows a disc blended with BisEDT (BE). [GM = gentamicin, CZ = cefazoline, FEP = cefepime, IPM = imipenem, SAM = ampicillin / sulbactam], LVX = levofloxacin. Figure 2 shows the effect of BisEDT and antibiotics on biofilm formation. S. Epidermidis was grown for 48 hours at 37 ° C. in TSB + 2% glucose in polystyrene plates. Gatifloxacin (GF), clindamycin (CM), minocycline (MC), gentamicin (GM), vancomycin (VM), cefazolin (CZ), nafcillin (NC), and rifampicin (RP). The results were expressed as the average rate of variation of BPC (2-fold serial dilution step) with 0.25 μM BisEDT (n = 3). Grown in TSB + 2% glucose at 37 ° C. for 48 hours. Figure 2 shows the effect of BisEDT and antibiotics on the growth of epidermidis. Results are expressed as mean MIC (dilution step) variation with increasing BisEDT (n = 3). See the legend of FIG. 5 for the definition of antibiotic. In bone and metal samples of open fractures in an in vivo rat model with or without cefazolin antibiotic treatment in addition to the three BT formulations, Bis-EDT, MB-11 and MB-8-2 It is a bar graph which shows the detected S. aureus average level. The standard error of the average value is shown as an error bar. Animals that were euthanized early were not excluded from the analysis, but a sample of one animal in group 2 was excluded due to significant contamination.

  Certain embodiments of the invention disclosed herein are specific bismuth-thiol (BT) compounds provided herein (preferably BT having a volume average diameter of about 0.4 μm to about 5 μm). Powerful disinfection, antibacterial and / or antibiofilm against specific bacteria, including bacteria associated with a number of clinically significant infections, including infections that contain microparticles may contain bacterial biofilms It is based on the surprising discovery that it shows activity but no other specific BT compounds (even if provided as microparticles).

  Contrary to expectations, not all BT compounds are uniformly effective in a predictable manner against such bacteria, but instead show different efficacy depending on the target species. In particular, as described herein, a specific BT compound can be applied in a manner that can for the first time provide clinically relevant strategies for the management of bacterial infections, including bacterial biofilm infections, by non-limiting theory. Although preferably found to contain higher efficacy against gram-negative bacteria, including BT microparticles having a volume average diameter of about 0.4 μm to about 5 μm, other specific BT compounds (preferably (Including BT microparticles having a volume average diameter of about 0.4 μm to about 5 μm) was found to show greater efficacy against gram positive bacteria.

  In addition, as described in more detail below, certain embodiments of the invention described herein may have particle sizes (eg, from about 0.4 μm to about 5 μm as disclosed herein). It relates to the surprising advantages provided by a novel bismuth-thiol (BT) composition that can be produced in a preparation comprising a plurality of BT microparticles that are substantially monodisperse with respect to (having a volume average diameter). In certain of these and related embodiments, the particulate BT is not provided as a component of lipid vesicles or liposomes, such as multilamellar phosphocholine-cholesterol liposomes or other multilamellar or unilamellar liposome vesicles.

  Also disclosed herein, certain antibiotic antibacterial and anti-biofilms previously found to lack a strong therapeutic effect against such bacterial infections with respect to certain embodiments Efficacy by treating the infection with either one or more of these antibiotics and the selected BT compound, either simultaneously, or sequentially, in either order (eg, an infection site such as a natural surface) Has been found to be significantly enhanced (eg, can be increased in a statistically significant manner). In an approach that could not have been predicted prior to the disclosure of the present invention, certain BT compounds may be used herein in combination with certain antibiotics for antibacterial and / or anti-biofilm activity against certain bacterial species or strains. Provided synergistic or enhancing combinations can be provided. Certain BT / antibiotics / combinations act synergistically or potentiate against a particular bacterium, while other specific BT / antibiotics / combinations have such synergistic or potentiating antibacterial and The observation that the anti-biofilm activity could not be demonstrated demonstrates the unexpected nature of such combinations described in more detail below.

  According to these and related embodiments, the antibiotic and BT compound may be administered simultaneously or sequentially in either order and may be formed by a particular infectious agent (eg, gram-negative or gram-positive). Specific synergistic or potentiating combinations of one or more antibiotics disclosed herein and one or more BT compounds to treat a biofilm) are predictable (eg, simply Instead of exhibiting (additive) activity, it is instead unexpectedly synergistic or potent (eg, superphase) as a function of the selected antibiotic, the selected BT compound and the specifically identified target bacteria It is noteworthy that it worked in an (additive) manner.

  For example, by way of example and not limitation, in aspects of a wide variety of natural and / or artificial surfaces that are actually or potentially microbially infected, and further aspects of improved substantially monodisperse particulate BT formulations Either or both of the specific antibiotic compounds and the specific BT compounds disclosed herein in may exhibit a slight antibacterial effect when used alone against specific strains or species However, the combination of both antibiotic and BT compounds is larger (statistically significant difference) than the simple sum of the effects of each compound when used alone against the same strain or species. Exerts a strong antibacterial effect, thereby, according to non-limiting theory, the anti-antagonistic effect of BT on the efficacy of antibiotics and / or on the efficacy of BT Substance -BT synergistic effect (e.g., FICI ≦ 0.5) is believed to reflect or enhancing effect (e.g. 0.5 <FICI ≦ 1.0). Thus, each BT compound is not synergistic or potent with respect to each antibiotic, and each antibiotic is not synergistic or potent with respect to each BT compound. -BT synergy and BT-antibiotic potentiation are generally not predictable. Instead, according to certain embodiments disclosed herein, certain combinations that synergize or enhance antibiotics and BT compounds are surprisingly natural and / or artificial as described herein. It imparts a strong antibacterial effect against specific bacteria, such as an antibacterial effect on a biofilm formed by a specific bacterium in a specific environment, such as a surface, and even in a specific situation.

  That is, certain BTs comprising antibiotics that can act synergistically with at least one BT composition comprising at least one BT compound provided herein (FICI ≦ 0.5). Antibiotics that synergize with are described herein, and such synergy is in the presence of antibiotics but no BT compounds and / or BT compounds but no antibiotics. In some cases, it manifests as a detectable effect on a larger scale than the detectable effect (ie, in a statistically significant manner with respect to the appropriate control conditions). Similarly, certain BT-antibiotic combinations show potentiating properties (0.5 <FICI <1.0), such potentiating properties in the presence of antibiotics but no BT compounds, and When a BT compound is present but no antibiotic is present, it develops as a detectable effect on a larger scale than that which can be detected (ie, in a statistically significant manner with respect to the appropriate control conditions).

  Examples of such detectable effects include, in certain embodiments, (i) prevention of infection by bacterial pathogens, (ii) inhibition of cell viability or cell growth of substantially all planktonic cells of bacterial pathogens. , (Iii) inhibition of biofilm formation by bacterial pathogens, and (iv) inhibition of biofilm viability or biofilm growth of substantially all biofilm-type cells of bacterial pathogenesis, In other contemplated embodiments, without limitation, antibiotic-BT synergy is one or more other clinically important parameters, such as one or more antibiotics or other drugs or chemistry. Degree of resistance or susceptibility of bacterial pathogens to agents, sensitive to one or more chemical, physical or mechanical conditions (eg pH, ionic strength, temperature, pressure) The degree of resistance and susceptibility to pathogens and / or one or more biological agents (eg, viruses, other bacteria, biologically active polynucleotides, immune cells or immune cell products such as antibodies, cytokines, chemokines, degrading enzymes) One or more of a change in the degree of resistance (eg, a statistically significant increase or decrease) of a bacterial pathogen to a starting enzyme, a membrane disrupting protein, a free radical, such as a reactive oxygen species, etc. It may be manifested as a detectable effect.

  As provided herein, an antibiotic present when the effect of a synergistic or potentiating antibiotic-BT combination exceeds the simple sum of the effects observed in the absence of one component of the combination. Those of skill in the art will understand these and various other criteria that can determine the effect of a particular agent on structure, function, and / or activity of a bacterial population in order to confirm substance-BT synergy or potentiation. Wax (e.g., Coico et al. (Eds.), Current Protocols in Microbiology, 2008, John Wiley & Sons, Hoboken, NJ; Schwalbe et al. 007, CRC Press, Boca Raton, FL).

  For example, in certain embodiments, antibacterial effects as described herein are measured using various concentrations of candidate agents (eg, BT and antibiotics alone or in combination) to determine Eliopoulos et al. (Antibiotics). in Laboratory Medicine (Lorian, V., Ed.), pp. 330-96, Williams and Willkins, Baltimore, MD, Eliopoulos and Moellering, (1996) Anticoin concentration in the USA. Synergy may be determined by calculating the bactericidal concentration index (FBCI). Synergy may be defined as a FICI or FBCI index of less than 0.5 and antagonism greater than 4 (see, eg, Odds, FC (2003) Synergy, antagonists, and what the chemotherboards putwethethem. Antimicrobiological Chemotherapy 52: 1). Synergy is conventionally defined as a four-fold decrease in antibiotic concentration or using partial inhibitory concentration (FIC) as described, for example, by Hollander et al. (1998 Antimicrob. Agents Chemother. 42: 744). May be. In certain embodiments, synergy is obtained with a combination of two drugs (eg, antibiotic and BT composition) and is greater than if the concentration of the second drug is replaced with the first drug ( It may be defined as a combined effect (for example, in a statistically significant manner).

  Thus, as described herein, it will be appreciated that in certain preferred embodiments, the combination of BT and antibiotics synergizes when a FICI value of 0.05 or less is observed (Odds, 2003). Also as described herein, in other specific preferred embodiments, and according to non-limiting theory, a specific BT-antibiotic combination may represent such a highly synergistic possibility. It is disclosed that it can exhibit FICI values between 5 and 1.0, the synergy of which is non-optimal of at least one BT and at least one antibiotic exhibiting unilateral or mutually enhanced cooperative antimicrobial efficacy May be observed using concentration. Such an effect may be referred to herein as “enhanced” antibiotic activity or “enhanced” BT activity.

  According to certain embodiments, (i) a concentration (in a statistically significant manner) that is lower than the characteristic minimum inhibitory concentration (MIC) of BT for a given target microorganism (eg, a given strain or strain) (Ii) a characteristic IC50 (concentration that inhibits the growth of 50% of the microbial population; eg, Sothill et al., 1992 J Antimicrob Chemother 29 (2): 137) (statistics) The presence of at least one antibiotic at a concentration lower than the biofilm prophylactic concentration (BPC) of the antibiotic for a given target microorganism, and When an antimicrobial agent (eg, BT or antibiotic) is used at the same concentration in the absence of the other antibacterial agent (eg, antibiotic or BT) In respect antimicrobial effects observed, when the result in high (in a statistically significant technique) BT- antibiotic antimicrobial efficacy of substances and combinations, enhancing antibiotics and / or BT activity can be detected. In preferred embodiments, “enhanced” antibiotic and / or BT activity is present when a FICI value of less than 1.0 and greater than 0.5 is measured.

  As will be appreciated by those of skill in the art based on the present disclosure, in certain embodiments, synergistic or potentiating antibiotics and / or BT activity may be generated by methods known in the art, for example, to the addition of Loewwe. Models based on (eg, FIC index, Greco model), or models based on Bliss independence (eg, nonparametric and semiparametric models) or other methods known in the art described herein (eg, Melediadis et al., 2005 Medical Mycology 43: 133-152). Exemplary methods for measuring synergistic or potentiating antibiotics and / or BT activity in this way are described, for example, by Melediadis et al. , 2005 Medical Mycology 43: 133-152 and references cited therein (also Melediadis et al., 2002 Rev Med Microbial 13: 101-117; White et al., 1996 AntimicrobAg: 40 centimeters). 1914-1918; see Mouton et al., 1999 Antimicrob Agents Chemother 43: 2473-3478).

  Other specific embodiments show that BT compounds and antibiotics do not exhibit predictable (simply additive) activity in vivo, instead of selecting selected antibiotics, selected BT compounds, and infectious agents. Unexpectedly synergistic or potentiating (eg, superadditive; or where two or more such agents are present in combination as a function of one or more of the specifically identified target species including Even when acting in a manner that provides a greater effect (eg, in a statistically significant manner) than would be obtained if the concentration of the second drug was replaced with the first drug. One or more antibiotics disclosed herein that may exhibit a synergistic or potentiating effect in vivo for the treatment of infectious agents (eg, biofilms formed by Gram negative or Gram positive bacteria) Substance and one or more It contemplates specific combination of the BT compound. Therefore, according to these and related embodiments, in certain in vivo situations, FICI or FBCI values (measured in vitro) may not be readily available and instead synergistic or potentiating effects of BT-antibiotics It will be appreciated that may be determined in a manner obtained by a quantifiable measure of the infectious agent.

  For example, in one embodiment, such as the rat femur critical defect model of in vivo open fracture described in Example 11, observed after treatment with BT-antibiotic combination, compared to antibiotic treatment or BT compound alone. A statistically significant decrease in the number of bacteria is an indication of a synergistic or potentiating effect. Statistical significance can be determined using methods well known to those skilled in the art. In other specific embodiments, at least 5%, 10%, 20%, 30% of the number of bacteria observed in the injury after treatment with BT-antibiotic combination, compared to antibiotic treatment or BT compound alone, A decrease observed in this or other in vivo models of 40% or 50% is considered an indicator of synergistic or enhancing effects.

  Other exemplary indicators of in vivo infection include established methodologies developed to quantify the severity of infection, such as various wound scoring systems known to those skilled in the art (eg, European Wound Management Association (EWMA), Position Document: Identifying criteria for Wound Infection. London: See scoring system reviewed in MEP Ltd, 2005). Exemplary wound scoring systems that can be used in assessing the synergistic or potentiating activity of the BT-antibiotic combinations described herein include ASEPSIS (Wilson AP, J Hosp Infect 1995; 29 (2). Wilson et al., Lancet 1986; 1: 311-13), the Southampton Wound Assessment Scale (Baley IS, Karlan SE, Toyn K, et al. BMJ 1992; 304: 1992; 304: 1992; . Horan TC, Gaynes P, Martone WJ, et al. 1992 Infect Control Hosp Epidemiol 1992; 13: 606-08. In addition, recognized clinical indicators of wound healing known to skilled clinicians such as wound size, depth, granulation tissue status, infection, etc. in the presence or absence of BT compounds and / or antibiotics You may measure with. Thus, based on the present disclosure, to determine whether a BT composition-antibiotic combination changes in vivo wound healing (eg, increases or decreases in a statistically significant manner relative to the appropriate control) Various methods will be readily appreciated by those skilled in the art.

  In view of these and related embodiments, microbially infected natural and artificial surfaces, such as surfaces that support or contain bacterial biofilms, are treated with one or more BT compounds and optionally one or more antibiotic compounds, such as An effective amount of a composition or formulation comprising one or more synergistic antibiotics or one or more potentiating antibiotics provided herein (eg, in certain embodiments, a therapeutically effective amount) A great variety of methods for treating with are provided herein. Based on this disclosure, it will be appreciated that certain antibiotics previously considered ineffective for a given type of infection by those skilled in the art are contemplated herein for use in the treatment of the same type of infection. I want.

  Thus, certain embodiments contemplate compositions that include one or more BT compounds that are used as disinfectants. A disinfectant is a substance that kills or interferes with the growth of microorganisms and is typically applied to living tissue to distinguish classification from those typically applied to inanimate objects. (Goodman and Gilman's “The Pharmacological Basis of Therapeutics”, Seventh Edition, Gilman et al., Editorial, 1985, Book of G. G. Common examples of disinfectants are ethyl alcohol and iodine tincture. Disinfectants include disinfectants that kill microorganisms such as microbial pathogens.

  Certain embodiments described herein include one or more BT compounds and one or more antibiotic compounds (eg, synergistic and / or potentiating antibiotics provided herein). A composition comprising it may be contemplated. Antibiotics are known in the art and are typically drugs produced by compounds that are produced by one species of microorganism and kill another species of microorganism, or the same or similar chemical structure and mechanism of action. For example, drugs that destroy microorganisms in or on the surface of living organisms when applied topically. In particular, embodiments disclosed herein are those in which the antibiotic may belong to one of the following classes: aminoglycosides, carbapenems, cephalosporins, fluoroquinolones, glycopeptide antibiotics, lincosamides (eg , Clindamycin), penicillinase resistant penicillin, and aminopenicillin. In other words, antibiotics need not be limited, but oxacillin, piperacillin, cefuroxime, cefotaxime, cefepime, imipenem, aztreonam, streptomycin, tobramycin, tetracycline, minocycline, ciprofloxacin, levofloxacin, erythromycin, linezolid, capreomycin, , Isoniazid, ansamycin, carbacephem, monobactam, nitrofuran, penicillin, quinolone, sulfonamide, clofazimine, dapsone, capreomycin, cycloserine, ethambutol, etionamide, isoniazid, pyrazinamide, rifamupicin, rifamupine, rifabutin, streptomycin, streptomycin, streptomycin Namin, Chloramphenicol, Phospho Isin, fusidic acid, linezolid, metronidazole, mupirocin, platencimycin, quinupristin, darfopristin, rifaximin, thiamphenicol, tinidazole, aminoglycoside, β-lactam, penicillin, cephalosporin, carbapenem, fluroquinolone, ketolide, lincosamide, macrolide Oxazolidinone, streptogramin, sulfonamide, tetracycline, glycylcycline, methicillin, vancomycin, nafcillin, gentamicin, ampicillin, chloramphenicol, doxycycline, tobramycin, amikacin, arbekacin, gentamicin, kanamycin, neomycin, netilmycin, paromomycin, rhodomycin Streptomycin, Tobrama Mention may be made of isine, apramycin, clindamycin, gatigloxacin, aminopenicillin, and others known in the art. An overview of these and other clinically useful antibiotics is available and known to those of skill in the art (see, eg, Washington University School of Medicine, The Washington, United States, Medical Therapeutics, 32nd World E., 2000). and Haucers, AL, Antibiotic Basics for Clinicians, 2007 Lipcottot, Williams and Wilkins, Philadelphia, PA) and Wilkins, Philadelphia, PA.

  An exemplary class of antibiotics used with one or more BT compounds in certain embodiments disclosed herein can be found in Edson RS, Terrell CL. The aminoglycosides. Mayo Clin Proc. 1999 May; 74 (5): 519-28, an aminoglycoside antibiotic. This class of antibiotics inhibits bacterial growth by damaging bacterial protein synthesis through binding and inactivation of bacterial ribosomal subunits. In addition to such bacteriostatic properties, aminoglycosides also exhibit bactericidal effects through the disruption of cell walls of gram-negative bacteria.

  Aminoglycoside antibiotics include gentamicin, amikacin, streptomycin and the like and are generally considered useful for the treatment of Gram-negative bacteria, mycobacteria and other microbial pathogens, but the classification of resistant strains has been reported. Aminoglycosides are not absorbed from the gastrointestinal tract and are therefore generally not considered applicable to oral formulations. Amikacin, for example, is often effective against gentamicin-resistant strains, but is typically administered intravenously or intramuscularly and can induce pain in patients. In addition, the toxicity associated with aminoglycoside antibiotics such as amikacin can lead to kidney damage and / or irreversible hearing damage.

  Despite these properties, certain embodiments disclosed herein are synergistic to treat epithelial tissue surfaces at one or more locations, eg, along the oral cavity, gastrointestinal / gastrointestinal tract. Contemplates oral administration of the BT / antibiotic combination (eg, the antibiotic need not be limited to aminoglycosides). Also contemplated in other specific embodiments are described herein as bactericides that refer to preparations that kill or block the growth of microorganisms on the outer surface of an inanimate object. Use of the prepared compositions and methods.

  As also described elsewhere herein, BT compounds are described in Domenico et al. , 1997 Antimicrob. Agent. Chemother. 41 (8): 1697-1703, Domenico et al. , 2001 Antimicrob. Agent. Chemother. 45 (5): 1417-1421, and U.S. Pat. S RE 37,793, U.S. Pat. S. 6,248,371, 6,086,921, and 6,380,248 (see also, for example, US 6,582,719) (including preparation methods) It may be a composition comprising a bismuth or bismuth salt and a thiol (eg, —SH, or sulfhydryl) -containing compound. However, certain embodiments are not so limited and other BT compounds including bismuth or bismuth salts and thiol-containing compounds may be contemplated. The thiol-containing compound may include 1, 2, 3, 4, 5, 6 or more thiol (eg, -SH) groups. In preferred embodiments, the BT compound comprises bismuth associated with a thiol-containing compound via ionic bonds and / or as a coordination complex, while in some other embodiments, bismuth is present in the organometallic compound. May be associated with the thiol-containing compound via a covalent bond. However, certain contemplated embodiments explicitly exclude organometallic compounds, such as BT compounds, where bismuth is a compound found in a covalent linkage to an organic moiety.

＊ 硫黄含有化合物と三価錯体を形成するために用いられる反応体の化学量論比およびビスマスの公知の傾向に基づき、比較のために単一ビスマス原子に対する原子比を示す。示された原子比は、所定の調製物中の全ての化学種の正確な分子式でない場合がある。カッコ内の数字は、１種（またはそれを超える）チオール剤に対するビスマスの比である（例えば、Ｂｉ：チオール１／チオール２）。「ＣＰＤ」：化合物。 Exemplary BT compounds are shown in Table 1.

* Based on the stoichiometric ratio of the reactants used to form the trivalent complex with the sulfur-containing compound and the known tendency of bismuth, the atomic ratio for a single bismuth atom is shown for comparison. The indicated atomic ratios may not be the exact molecular formula of all chemical species in a given preparation. The numbers in parentheses are the ratio of bismuth to one (or more) thiol agent (eg, Bi: thiol 1 / thiol 2). “CPD”: Compound.

  The BT compounds used in certain of the embodiments disclosed herein are well-established procedures (eg, US RE 37,793, US 6,248,371, 6,086, 921, and 6,380,248; Domenico et al., 1997 Antimicrob. Agent. Chemother. 41 (8): 1697-1703, Domenico et al., 2001 Antimicrob. Agent. 1417-1421), and in other specific embodiments, BT compounds may be prepared according to the methodology described herein. That is, certain preferred embodiments may be used under conditions and times sufficient to form a precipitate comprising microparticles comprising a BT compound (eg, concentrations, solvents, as described herein and understood by those of skill in the art based on this disclosure). (Under conditions such as strength, temperature, pH, mixing and / or pressure), the dissolved bismuth is at least 50 mM, at least 100 mM, at least 150 mM, at least 200 mM, at least 250 mM, at least 300 mM, at least 350 mM, at least 400 mM, at least 500 mM, at least An acidic bismuth aqueous solution containing 600 mM, at least 700 mM, at least 800 mM, at least 900 mM, or at least 1 M and containing no hydrophilic, polar or organic solubilizer is mixed with ethanol to form a first ethanol. BT is obtained and reacted with a second ethanolic solution containing a thiol-containing compound present in the reaction solution in a molar ratio of about 1: 3 to about 3: 1 with respect to bismuth to obtain a reaction solution. The synthetic methods described herein for preparing compounds, and in particular for obtaining BT compounds in substantially monodisperse particulate form, are contemplated.

  Thus, exemplary BTs include Compound 1B-1, Bis-EDT (1: 1 bismuth-1,2-ethanedithiol reactant); Compound 1B-2, Bis-EDT (1: 1.5); Compound 1B -3, Bis-EDT (1: 1.5); Compound 1C, Bis-EDT (soluble Bi preparation, 1: 1.5); Compound 2A, Bis-Bal (bismuth-British anti-Lewisite (bismuth-di) Mercaprol, bismuth-2,3-dimercaptopropanol), 1: 1); Compound 2B, Bis-Bal (1: 1.5); Compound 3A Bis-Pyr (bismuth pyrithione, 1: 1.5); Compound 3B, Bis-Pyr (1: 3); Compound 4, Bis-Ery (bismuth dithioerythritol, 1: 1.5); Compound 5, Bis-Tol (bismuth- 3,4-dimercaptotoluene, 1: 1.5); Compound 6, Bis-BDT (bismuth-2,3-butanediol, 1: 1.5); Compound 7, Bis-PDT (bismuth-1,3) -Propanedithiol, 1: 1.5); Compound 8-1 Bis-Pyr / BDT (1: 1/1); Compound 8-2, Bis-Pyr / BDT (1: 1 / 0.5); Compound 9 Bis-2-hydroxy, propanethiol (bismuth-1-mercapto-2-propanol, 1: 3); Compound 10, Bis-Pyr / Bal (1: 1 / 0.5); Compound 11, Bis-Pyr / EDT (1: 1 / 0.5); Compound 12 Bis-Pyr / Tol (1: 1 / 0.5); Compound 13, Bis-Pyr / PDT (1: 1 / 0.5); Compound 14 Bis- Pyr / ry (1: 1 / 0.5); compound 15, Bis-EDT / 2- hydroxy propane thiol (1: 1/1) and the like (e.g., see Table 1).

  While not wishing to be bound by theory, in certain preferred embodiments, the BT may comprise preparing or obtaining an acidic aqueous liquid solution containing bismuth, such as an aqueous nitric acid solution containing bismuth nitrate. The methods of the present disclosure for preparing the compounds can, if desired, facilitate ease of mass production, improved product purity, homogeneity or consistency (including particle size uniformity), or topical formulations of the present invention. It is contemplated that a composition comprising a BT compound can be obtained having one or more desired properties, including other properties useful for the preparation and / or administration of

  In certain embodiments, the BT compositions prepared by the methods described herein for the first time each have a volume average diameter (VMD) of about 0.4 μm to about 5 μm, according to certain presently preferred embodiments. It has been discovered that it exhibits advantageous homogeneity in relation to their appearance as a substantially monodispersed particulate suspension. The measured particle diameter can be referred to as the volume average diameter (VMD), mass median diameter (MMD), or aerodynamic mass median diameter (MMAD). These measurements may be performed, for example, by characterization with impaction (MMD and MMAD) or laser (VMD). In the case of liquid particles, VMD, MMD, and MMAD can be the same if environmental conditions are maintained, eg, standard humidity. However, if the humidity is not maintained, the MMD and MMAD measurements will be smaller than the VMD due to dehydration during the impactor measurement. For purposes of this specification, VMD, MMD and MMAD measurements will be considered under standard conditions, so the description of VMD, MMD and MMAD would be equivalent. Similarly, dry powder particle size measurements with MMD and MMAD are considered equivalent.

  As described herein, a preferred embodiment relates to a substantially monodispersed suspension of BT-containing microparticles. Generating a defined BT particle size with a slight geometric standard deviation (GSD) can, for example, attach BT, accessibility to a desired target site in or on a natural surface, and / or BT microparticles Tolerability may be optimized by the subject to whom is administered. A narrow GSD will limit the number of particles outside the desired VMD or MMAD size range.

  In one embodiment, a liquid or aerosol suspension of particulates having a VMD of about 0.5 microns to about 5 microns containing one or more of the BT compounds disclosed herein. In another embodiment, a liquid or aerosol suspension having a VMD or MMAD of about 0.7 microns to about 4.0 microns is provided. In another embodiment, a liquid or aerosol suspension having a VMD or MMAD of about 1.0 microns to about 3.0 microns is provided. In other specific embodiments, the VMD is about 0.1 to about 5.0 microns, or about 0.1, about 0.2, about 0.3, about 0.4, about 0.5, about 0.0. 6, about 0.7, about 0.8, or about 0.9 to about 1.0, about 1.5, about 2.0, about 2.5, about 3.0, about 3.5, about 4. 0, about 4.5, about 5.0, about 5.5, about 6.0, about 6.5, about 7.0, about 7.5, or about 8.0 microns described herein A liquid suspension comprising one or more BT compound particles prepared as described above is provided.

  Thus, in certain preferred embodiments, “substantially” all of the BT preparations first described herein that are “substantially” monodisperse, eg, “substantially” all of the microparticles are within a specific range (eg, A BT composition comprising a BT compound in particulate form having a volume average diameter (VMD) of .4 μm to about 5 μm) is at least 80%, 85%, 90%, 91%, 92%, 93%, or 94 of the particles. %, More preferably at least 95%, 96%, 97%, 98%, 99% or more of those compositions having VMD within the quoted size range.

  These and related properties of BT compositions prepared by the synthetic methods described herein enable characterization in a manner that promotes regulatory compliance in accordance with one or more pharmaceutical, prescription and medicinal cosmetic standards. Presents unprecedented advantages over previously described BT, such as lower manufacturing costs and ease of manufacture, and homogeneity within the composition.

  In addition or alternatively, the substantially monodispersed BT microparticles described herein advantageously do not require micronization, i.e. expensive and laborious milling or supercritical fluid processing, or microparticles May be produced without using other equipment and procedures typically used to generate (eg, Martin et al. 2008 Adv. Drug Deliv. Rev. 60 (3): 339; Moribe et al. Rev. 60 (3): 328; Cape et al., 2008 Pharm.Res. 25 (9): 1967; Rasenack et al. 2004 Pharm.Dev.Technol. 1-13). Thus, embodiments of the present invention include, but are not limited to, enhanced and substantially homogeneous solubilizing properties, suitability for desired dosage forms such as oral, inhalation or topical forms on dermatological / skin wounds, It presents the beneficial effects of a substantially homogeneous microparticle preparation, including a high degree of bioavailability and other beneficial properties.

  BT compound particulate suspensions are, for example, wetting agents, surfactants, mineral oils, or other ingredients or additives that would be known to those skilled in the formulation to retain individual particulates in the suspension. As an aqueous formulation, as a preparation containing, as a suspension or solution in an aqueous and organic solvent such as a halogenated hydrocarbon propellant, as a dry powder, or in other forms detailed below be able to. Aqueous formulations may be aerosolized with a liquid nebulizer, for example, using either a hydraulic or ultrasonic spray. A propellant-based system may use a suitable pressurized dispenser. As the dry powder, a dry powder dispersion device capable of effectively dispersing the BT-containing fine particles may be used. The desired particle size and distribution may be obtained by selecting an appropriate device.

  As previously mentioned, also provided herein by certain embodiments is a method of preparing a BT composition comprising a plurality of microparticles comprising a bismuth-thiol (BT) compound, such as Substantially all of the fine particles have a volume average diameter (VMD) of from about 0.1 to about 8 microns, and in certain preferred embodiments from about 0.4 microns to about 5 microns.

  In general conditions, the method comprises (a) a bismuth salt containing bismuth at a concentration of at least 50 mM, under conditions and time sufficient to obtain a solution substantially free of solid precipitate, (Ii) admixed with a sufficient amount of ethanol to mix at least about 5%, 10%, 15%, 20%, 25% Or obtaining an admixture comprising 30% by volume, preferably about 25% by volume ethanol; and (b) about 1: 3 to bismuth with conditions and time sufficient to form a precipitate comprising the BT compound. Adding an ethanolic solution containing a thiol-containing compound present in the reaction solution in a molar ratio of about 3: 1 to the blend of (a) to obtain a reaction solution.

  In certain preferred embodiments, the bismuth salt may be Bi (NO 3) 3, but it will be understood by the present disclosure that bismuth can be provided in other forms. In certain embodiments, the bismuth concentration in the acidic aqueous solution is at least 100 mM, at least 150 mM, at least 200 mM, at least 250 mM, at least 300 mM, at least 350 mM, at least 400 mM, at least 500 mM, at least 600 mM, at least 700 mM, at least 800 mM, at least 900 mM or It may be at least 1M. In certain embodiments, the acidic aqueous solution comprises at least 5 wt%, 10 wt%, 15 wt%, 20 wt%, 22 wt% or 22.5 wt% bismuth. The acidic aqueous solution may comprise at least 5 wt% or more nitric acid in certain preferred embodiments, and in other specific embodiments, the acidic aqueous solution is at least 0.5 wt%, at least 1 wt%. At least 1.5 wt%, at least 2 wt%, at least 2.5 wt%, at least 3 wt%, at least 3.5 wt%, at least 4 wt%, at least 4.5 wt% or at least 5 wt% nitric acid May be included.

  The thiol-containing compound may be any thiol-containing compound described herein, and in certain embodiments, 1,2-ethanedithiol, 2,3-dimercaptopropanol, pyrithione, dithioerythritol, Including one or more of 3,4-dimercaptotoluene, 2,3-butanedithiol, 1,3-propanedithiol, 2-hydroxypropanethiol, 1-mercapto-2-propanol, and dithiothreitol Also good. Other exemplary thiol-containing compounds include α-lipoic acid, methanethiol (CH3SH [m-mercaptan]), ethanethiol (C2H5SH [e-mercaptan]), 1-propanethiol (C3H7SH [n-P mercaptan]). ), 2-propanethiol (CH3CH (SH) CH3 [2C3 mercaptan]), butanethiol (C4H9SH [n-butyl mercaptan]), tert-butyl mercaptan (C (CH3) 3SH [t-butyl mercaptan]), pentanethiol (C5H11SH [pentyl mercaptan]), coenzyme A, lipoamide, glutathione, cysteine, cystine, 2-mercaptoethanol, dithiothreitol, dithioerythritol, 2-mercaptoindole, transglutaminase, o And any of the following thiol compounds available from Sigma-Aldrich (St. Louis, MO): (11-mercaptoundecyl) hexa (ethylene glycol), (11-mercaptoundecyl) tetra (ethylene glycol), ( 11-mercaptoundecyl) tetra (ethylene glycol) functionalized gold nanoparticles, 1,1 ′, 4 ′, 1 ″ -terphenyl-4-thiol, 1,11-undecanedithiol, 1,16-hexadecanedithiol Technical grade 1,2-ethanedithiol, 1,3-propanedithiol, 1,4-benzenedimethanethiol, 1,4-butanedithiol, 1,4-butanedithioldiacetate, 1,5- Pentanedithiol, 1,6-hexanedithiol, 1,8-octanedithio All, 1,9-nonanedithiol, adamantanethiol, 1-butanethiol, 1-decanethiol, 1-dodecanethiol, 1-heptanethiol, 1-heptanethiolpurum, 1-hexadecanethiol, 1-hexanethiol, 1- Mercapto (triethylene glycol), 1-mercapto (triethylene glycol) methyl ether functionalized gold nanoparticles, 1-mercapto-2-propanol, 1-nonanethiol, 1-octadecanethiol, 1-octanethiol, 1-pentadecane Thiol, 1-pentanethiol, 1-propanethiol, 1-tetradecanethiol, 1-tetradecanethiolpurum, 1-undecanethiol, 11- (1H-pyrrol-1-yl) undecane-1-thiol, 11-amino-1 -Eun Canthiol hydrochloride, 11-bromo-1-undecanthiol, 11-mercapto-1-undecanol, 11-mercaptoundecanoic acid, 11-mercaptoundecyl trifluoroacetate, 11-mercaptoundecyl phosphate, 12-mercaptododecane Acid, 15-mercaptopentadecanoic acid, 16-mercaptohexadecanoic acid, 1H, 1H, 2H, 2H-perfluorodecanethiol, 2,2 '-(ethylenedioxy) diethanethiol, 2,3-butanedithiol, 2-butane Thiol, 2-ethylhexanethiol, 2-methyl-1-propanethiol, 2-methyl-2-propanethiol, 2-phenylethanethiol, 3,3,4,4,5,5,6,6,6- Nonafluoro-1-hexanethiolpurum, 3- (dimethoxyme Rusilyl) -1-propanethiol, 3-chloro-1-propanethiol, 3-mercapto-1-propanol, 3-mercapto-2-butanol, 3-mercapto-N-nonylpropionamide, 3-mercaptopropionic acid, 3 -Mercaptopropyl functionalized silica gel, 3-methyl-1-butanethiol, 4,4'-bis (mercaptomethyl) biphenyl, 4,4'-dimercaptostilbene, 4- (6-mercaptohexyloxy) benzyl alcohol, 4-cyano-1-butanethiol, 4-mercapto-1-butanol, 6- (ferrocenyl) hexanethiol, 6-mercapto-1-hexanol, 6-mercaptohexanoic acid, 8-mercapto-1-octanol, 8-mercapto Octanoic acid, 9-mercapto-1-nonano , Biphenyl-4,4'-dithiol, butyl 3-mercaptopropionate, copper (I) 1-butanethiolate, cyclohexanethiol, cyclopentanethiol, decanethiol functionalized silver nanoparticles, dodecanethiol functionalized gold Nanoparticles, dodecanethiol functionalized silver nanoparticles, hexa (ethylene glycol) mono-11- (acetylthio) undecyl ether, mercaptosuccinic acid, methyl 3-mercaptopropionate, NanoTether BPA-HH, NanoThink ™ 18, NanoThinks (TM) 8, NanoThinks (TM) ACID11, NanoThinks (TM) ACID16, NanoThinks (TM) ALCO11, NanoThinks (TM) THIO8, Octanethio Functionalized gold nanoparticles, PEG dithiol average Mn 8,000, PEG dithiol average molecular weight 1,500, PEG dithiol average molecular weight 3,400, S- (11-bromoundecyl) thioacetate, S- (4-cyanobutyl) thioacetate, Thiophenol, triethylene glycol mono-11-mercaptoundecyl ether, trimethylolpropane tris (3-mercaptopropionate), [11- (methylcarbonylthio) undecyl] tetra (ethylene glycol), m-carborane-9- Thiol, p-terphenyl-4,4 "-dithiol, tert-dodecyl mercaptan and tert-nonyl mercaptan.

  Exemplary reaction conditions such as temperature, pH, reaction time, use of stirring or agitation to dissolve solutes, and procedures for recovering and washing the precipitate are described herein and are generally described in the art. A known technique is used.

  Unlike previously described methodologies for producing BT compounds, according to the method of the present invention for preparing BT, the BT product, in certain preferred embodiments, is about 0.4 microns to about Provided as a microparticle suspension with substantially all microparticles having a VMD of 5 microns, and according to other particular embodiments, generally from about 0.1 microns to about 8 microns. Further, unlike previous approaches, according to embodiments of the present invention, bismuth is a bismuth salt at a concentration of at least about 50 mM to about 1 M, and at least about 0.5% (w / w) to about 5% (w / W), preferably in an acidic aqueous solution containing nitric acid in an amount of less than 5% (w / w) and free of hydrophilic, polar or organic solubilizers.

  In this regard, the method of the present invention is such that bismuth is 50 μM and is not water soluble (eg, US RE 37793) and bismuth is unstable in water (eg, Kuvshinova et al., 2009 Russ. J). Inorg. Chem 54 (11): 1816), surprising and expected in view of the generally recognized art that bismuth is also unstable in aqueous nitric acid unless hydrophilic, polar or organic solubilizers are present. Present outside profits. For example, in all of the most reliable descriptions of BT preparation methodologies (eg, Domenico et al., 1997 Antimicrob. Agents. Chemother. 41: 1697; US 6,380,248; US RE 37,793). No., US 6,248,371), the hydrophilic solubilizing agent propylene glycol is required to dissolve bismuth nitrate and the bismuth concentration of the solution prepared for reaction with thiol is 15 mM. Much lower, thereby limiting the available modes of production of BT compounds.

  In contrast, according to the present disclosure, a hydrophilic, polar or organic solubilizer is not required to dissolve bismuth, and surprisingly higher concentrations are achieved. Hydrophilic, polar or organic solubilizers include propylene glycol (PG) and ethylene glycol (EG), polar solvents such as dioxane and dimethylene sulfoxide (DMSO), polyols (eg polyethylene glycol (PEG), polypropylene) (Including glycols (PPG), pentaerythritol, etc.), polyhydric alcohols such as glycerol and mannitol, and any of a number of known solubility enhancers, including other drugs. Other highly polar water-miscible organic materials include dimethyl sulfoxide (DMSO), dimethylformamide (DMF) and NMP (N-methyl-2-pyrrolidone).

  Thus, solvents such as those commonly used as hydrophilic, polar or organic solubilizers provided herein include, for example, Catalan et al. (Eg, 1995 Liebigs Ann. 241; Handbook of Solvent, Wypych (Ed.), May also be selected based on the solvent polarity / polarizability (SPP) scale values using the system of Andrew Publ., Catalan in NY, 2001, and references cited therein, For example, those skilled in the art will appreciate that water has an SPP value of 0.962, toluene has an SPP value of 0.655, and 2-propanol has an SPP value of 0.848. A method for measuring the SPP value of a solvent based on UV measurement of 2-N, N-dimethyl-7-nitrofluorene / 2-fluoro-7-nitrofluorene probe / homophoric pair has been described ( Catalan et al., 1995).

  Solvents with the desired SPP value based on the solubility characteristics of the particular BT composition (either as a pure single component solvent or a solvent mixture of two, three, four or more solvents; solvent miscibility Regarding gender, for example, Godfrey 1972 Chem. Technol. 2: 359) can be readily identified by one of ordinary skill in the art in view of the present disclosure, but as noted above, the synthesis described herein According to certain preferred embodiments for the method steps, no hydrophilic, polar or organic solubilizer is required to dissolve bismuth.

  Solubility parameters may include interaction parameter C, Hildebrand solubility parameter d, or partial (Hansen) solubility parameters: δp, δh and δd, representing solvent polarity, hydrogen bonding ability and dispersive force interaction ability, respectively. . In certain embodiments, the highest solubility parameter describing the solvent or co-solvent system in which the bismuth salt comprising bismuth dissolves is obtained, for example, by the method of preparing the particulate BT composition described herein. Limits of aqueous solution containing may be provided. For example, a higher δh value has a greater hydrogen bonding capacity and thus a greater affinity for solvent molecules such as water. Therefore, in situations where a more hydrophilic environment is desired, it may be preferred that the observed solvent maximum δh be a higher value.

As a non-limiting example, BisEDT having the structure shown in Formula I below may be prepared according to the following reaction scheme:

  Briefly, as a non-limiting example, an excess of 5% aqueous HNO3 at room temperature (11.4 L) is added to a Bi (NO3) 3 solution (eg, 43% Bi (NO3) 3 (w / w), 5 Slowly add 0.331 L (about 0.575 mol) of acidic bismuth aqueous solution such as% nitric acid (w / w), 52% water (w / w), obtained from Shepherd Chemical Co., Cincinnati, OH Then, absolute ethanol (4 L) may be gradually added. Separately, 72.19 mL (0.863 mol) of 1,2-ethanedithiol was added to 1.5 L of absolute ethanol using a 60 mL syringe, followed by stirring for 5 minutes, thereby allowing 1,2-ethanedithiol [˜0. An ethanolic solution (1.56 L) of a thiol compound such as .55M] may be prepared. 1,2-ethanedithiol (CAS 540-63-6) and other thiol compounds are described, for example, in Sigma-Aldrich, St. Available from Louis, MO. Thereafter, an ethanolic solution of the thiol compound may be gradually added to the aqueous Bi (NO3) 3 / HNO3 solution and stirred overnight to form a reaction solution. The thiol-containing compound may be present in the reaction solution in a molar ratio of about 1: 3 to about 3: 1 with respect to bismuth, according to certain preferred embodiments. The product formed is precipitated as a precipitate containing the particulates described herein, then collected by filtration and subsequently washed with ethanol, water and acetone to give BisEDT as a yellow amorphous powdered solid. obtain. The crude product may be redissolved in absolute alcohol with stirring, then filtered and subsequently washed several times with ethanol and then several times with acetone. The washed powder may be triturated in 1M NaOH (500 mL), filtered and subsequently washed with water, ethanol and acetone to obtain purified microparticle BisEDT.

  According to a non-limiting theory, bismuth inhibits the ability of bacteria to produce extracellular polymeric substances (EPS), such as extracellular polysaccharides, which leads to impaired biofilm formation. Bacteria are thought to use glue-like EPS for biofilm adhesion. Depending on the nature of the infectious agent, biofilm formation and EPS assimilation may contribute to bacterial pathogenicity such as interfering with wound healing. However, bismuth alone is not therapeutically useful as an intervention agent and is instead typically administered as part of a complex such as BT. Thus, bismuth-thiol (BT) is a family of compositions comprising compounds obtained from chelation of bismuth with thiol compounds and exhibiting dramatic improvements in the antimicrobial therapeutic efficacy of bismuth. BT exhibits a pronounced anti-infective effect, anti-biofilm effect, and immune modulating effect. Bismuth-thiol is effective against a wide range of microorganisms and is typically not affected by antibiotic resistance. BT can prevent biofilm formation at significantly lower (sub-inhibitory) concentrations, prevent many pathogenic features of common wound pathogens at the same sub-inhibitory level, and prevent septic shock in animal models; Can be synergistic with many antibiotics.

  As described herein, such synergy in the antibacterial effect of one or more specific BTs when combined with one or more specific antibiotic compounds is a distinct antibiotic for a particular bacterial type. And a BT effect profile that cannot be predicted immediately, but surprisingly, certain specific bacterial populations, such as confirming the presence of Gram-negative or Gram-positive bacteria (or both), are considered. It may be obtained by selecting a BT-antibiotic / combination. For example, as disclosed herein, antibiotics that synergize with a particular BT include amikacin, ampicillin, aztreonam, cefazolin, cefepime, chloramphenicol, ciprofloxacin, clindamycin (or other lincosamide antibiotics) Substance), daptomycin (Cubicin®), doxycycline, gatifloxacin, gentamicin, imipenim, levofloxacin, linezolid (Zyvox®), minocycline, nafcillin, paromomycin, rifampin, sulfamethoxazole, tetracycline, bramycin And one or more of vancomycin may be included. For example, MRSA, which is individually or not at all sensitive to gentamicin, cefazoline, cefepime, sfamethoxazole, imipenim or levofloxacin, is exposed to these antibiotics in the presence of the BT compound BisEDT. It has been shown in an in vitro test that it exhibits a marked sensitivity to one or the other. Thus, specific embodiments contemplated herein include BT compounds and amikacin, ampicillin, cefazolin, cefepime, chloramphenicol, ciprofloxacin, clindamycin (or other lincosamide antibiotics), daptomycin (Cubicin®), doxycycline, gatifloxacin, gentamicin, imipenim, levofloxacin, linezolid (Zyvox®), minocycline, nafcillin, paromomycin, rifampine, sulfamethoxazole, tobramycin and vancomycin Explicitly contemplated compositions and / or methods that may include combinations with one or more antibiotics, other specific embodiments contemplated herein include BT compounds, amikacin, Picillin, aztreonam, cefazolin, cefepime, chloramphenicol, ciprofloxacin, clindamycin (or other lincosamides), daptomycin (Cubicin®), doxycycline, gatifloxacin, gentamicin, imipenim, levofloxacin, One or more antibiotics from which one or more antibiotics selected from linezolid (Zyvox®), minocycline, nafcillin, paromomycin, rifampine, sulfamethoxazole, tobramycin and vancomycin can be explicitly excluded; Compositions and / or methods that may include combinations of these are contemplated. It should be noted herein that gentamicin and tobramycin belong to the antibiotic aminoglycosides. Also explicitly excluded from certain contemplated embodiments are Domenico et al. , 2001 Agent. Chemother. 45: 1417-1421; Domenico et al. 2000 Infect. Med. 17: 123-127; Antimicrob. Agents & Chemother. 3: 79-85 in Domenico et al. , 2003 Res. Adv. Domenico et al. , 1997 Antimicrob. Agents Chemother. 41 (8): 1697-1703; Domenico et al. 1999 Infect. Immun. 67: 664-669; Huang et al. 1999 J Antimicrob. Chemother. 44: 601-605; Veloirah et al. , 2003 J Antimicrob. Chemother. 52: 915-919; Wu et al. , 2002 Am J Respir Cell Mol Biol. 26: 731-738; Halwani et al. , 2008 Int. J Pharmaceut. 358: 278; Halwani et al. , 2009 Int. J. et al. Pharmaceut. It is noted that none of these publications teach or suggest the monodisperse particulate BT composition disclosed herein. I want.

  Thus, as described herein, in certain preferred embodiments, the microparticles BT described herein are included, and optionally in other specific embodiments, synergistic and / or potentiating antibiotics are also included. Compositions and methods for treating a plant, animal or human subject, or product with a composition comprising are provided. One of ordinary skill in the relevant arts will be aware of appropriate agricultural, clinical, commercial, industrial, manufacturing, household, and other aspects and situations where such treatment may be desirable based on the present disclosure. The criteria include, inter alia, geriatric, cardiovascular, metabolic disease (eg, diabetes, obesity, etc.), infection and inflammation (respiratory or respiratory, eg, surgical, military surgical, dermatological, trauma care Established in the medical technology field, including the epithelial lining of the gastrointestinal tract, or other epithelial tissue surfaces such as glandular tissue), and related medical and sub-specialty areas.

  Preferred compositions for treating microbial infections on or within natural or artificial surfaces used in accordance with the embodiments described herein are, in certain embodiments, bismuth-thiol (BT) described herein. A composition comprising a compound, which, in particular but related embodiments, includes other compounds known in the art, such as one or more antibiotic compounds described herein. You may go out. BT compounds and methods for making them are disclosed herein, see, for example, Domenico et al. (1997 Antimicrob. Agent. Chemother. 41 (8): 1697-1703; 2001 Antimicrob. Agent. Chemother. 45 (5): 1417-1421) and U.S. Pat. S RE 37,793, U.S. Pat. S. 6,248,371, 6,086,921, and 6,380,248. As also mentioned earlier, certain preferred BT compounds include those that are ionically bound to a thiol-containing compound or that contain bismuth or a bismuth salt in a coordination complex therewith, such as bismuth chelated to a thiol-containing compound. Other particularly preferred BT compounds are those containing bismuth or a bismuth salt in a covalent linkage to a thiol-containing compound. Also preferred are the substantially monodispersed particulate BT compositions described herein. Described for the first time herein, having use in compositions and methods for promoting treatment described herein, either from past efforts to treat bacterial infections, or natural and / or artificial surfaces. Characterization of other aspects of any given compound also did not predict that the methods of the invention using such compounds would have the beneficial effects described herein.

  Thus, according to a preferred embodiment, there is provided a method of treating a natural or artificial surface comprising administering to the natural surface at least one particulate BT compound described herein. In certain embodiments, the method may be a synergistic antibiotic described herein in certain preferred embodiments, and may be described herein in other particular embodiments. The method further includes administering at least one antibiotic compound, which may be a potent antibiotic, simultaneously or sequentially in either order. Antibiotic compounds include aminoglycoside antibiotics, carbapenem antibiotics, cephalosporin antibiotics, fluoroquinolone antibiotics, glycopeptide antibiotics, lincosamide antibiotics, penicillinase-resistant penicillin antibiotics, or aminopenicillin antibiotics It may be an antibiotic. Clinically useful antibiotics are discussed elsewhere in this specification, for example, Washington University School of Medicine, The Washington Manual of Medical Therapeutics (32nd Ed.). , PA; Hauser, AL, Antibiotic Basics for Clinicians, 2007 Lippincott, Williams and Wilkins, Philadelphia, PA.

  As described herein, certain embodiments show that in the case of bacterial infections including gram positive bacteria, the preferred therapeutically effective formulation is a BT compound (eg, BisEDT, bismuth; 1,2- Ethanedithiol; BisPyr, bismuth: pyrithione; BisEDT / Pyr, bismuth: 1,2-ethanedithiol / pyrithione) and rifamycin, or BT compound and daptomycin (Cubicin®, Cubist Pharmaceuticals, Lxinton, MA), or BT Compound and linezolid (Zyvox®, Pfizer, Inc., NY, NY), or BT compound (eg, BisEDT, bismuth; 1,2-ethanedithiol; BisPyr, bismuth: pyrithione; BisEDT / P r, bismuth: 1,2-ethanedithiol / pyrithione) and ampicillin, cefazolin, cefepime, chloramphenicol, clindamycin (or other lincosamide antibiotics), daptomycin (Cubicin®), doxycycline, gatiflo The unexpected discovery that it may include one or more of xacin, gentamicin, imipenim, levofloxacin, linezolid (Zyvox®), nafcillin, paromomycin, rifampine, sulfamethoxazole, tobramycin and vancomycin Derived from.

  As also described herein, certain embodiments provide the unexpected that in the case of bacterial infections including gram-negative bacteria, a preferred therapeutically effective formulation may include a BT compound and amikacin. Derived from discovery. Certain related embodiments contemplate treating an infectious agent comprising a Gram-negative bacterium with a BT compound and another antibiotic, such as another aminoglycoside antibiotic that is not gentamicin or tobramycin in certain embodiments. Thus, in view of these embodiments, other related embodiments are described in the medical methodology arts as a step of selecting an appropriate antibiotic compound to be included in a formulation administered by the methods of the present invention. By methodology well known to those skilled in the art, it is contemplated to identify one or more bacterial populations or subpopulations within or on a natural or artificial surface by well-known criteria that are Gram positive or Gram negative.

  The compositions and methods described herein typically involve the compounds described herein (eg, one or more microparticles BT alone, or one or more synergisticities disclosed herein). And / or in combination with potentiating antibiotics) by applying or administering to microbial sites such as the presence of microorganisms on or in natural or artificial surfaces (eg bacteria, viruses, yeasts, molds). And other fungi, microbial parasites, etc.) may find use. Such natural surfaces include, but are not limited to, all surfaces of plants (eg, roots, bulbs, stems, leaves, branches, vines, toothpicks, buds, flowers or parts thereof, shoots, fruits, seeds, buds, etc. Or part), mammalian tissue (eg, skin, scalp, lining of the digestive tract, oral cavity, etc .; endothelium, cells and tissue membranes, such as peritoneum, pericardium, capsule, periosteum, meninges, myofibular membranes; And other mammalian tissues such as muscle, heart, lung, kidney, liver, spleen, gallbladder, pancreas, bladder, nerve, tooth, bone, joint, tendon, ligament, etc.) Any part of the product where the presence of microorganisms can be found (eg, walls, windows, floors, underfloor spaces, attics, basements, fences, roofs of commercial, residential, industrial, educational, health care and other public buildings) , Ceilings, lighting and piping fixtures, vents, ducts, pipelines, door knobs, Switches, sanitary equipment, drainage equipment, tanks, water pipes; medical and dental equipment, implants, tools, equipment, equipment, etc .; metal, glass, plastic, wood, rubber and paper products; transport equipment, eg shipping containers, automobiles, Railway equipment, boats, ships (eg external hulls, rudder, anchor and / or propeller surfaces, internal fixation devices and ballast tanks, and other internal surfaces), barges, and other marine equipment such as docks, bulkheads, wharves, etc. Also mentioned.

  The particulate antimicrobial agent described herein treats products, including natural and / or artificial surfaces, to suppress microbial growth, reduce microbial infection, and product resistance to microbial infection. Improve, reduce biofilm, prevent bacterial to biofilm conversion, prevent or inhibit microbial infection, prevent damage, and any other described herein May be used for These agents include a number of antivirals, including viral infection by herpes viruses such as cytomegalovirus, herpes simplex type 1 and herpes simplex type 2 and / or prevention or inhibition of infection by other viruses. It is also useful for purposes. In this context, the drugs are various viruses such as single stranded RNA viruses, single stranded DNA viruses, Rous sarcoma virus (RSA), hepatitis A virus, hepatitis B virus (HBV), hepatitis C. Virus (HCV), influenza virus, West Nile virus (WNV), Epstein-Barr virus (EBV), Eastern equine encephalitis virus (EEEV), severe acute respiratory virus (SARS), human immunodeficiency virus (HIV), human papilloma Virus (HPV), and human T cell lymphoma virus (HTLV) and plant pathogens (eg potato leaf curl virus; potato virus A, M, S, X or Y; tomato yellow gangrene virus; grape leaf associated virus 3; plum Poxvirus; lettuce mosaic virus; Pino mosaic virus; pepper mild mottle virus; useful in the prevention or inhibition of Impatiens necrotic mottle virus, etc.) as a viral infection viruses, including known, tomato mosaic virus; tobacco mosaic virus; Caribbean et core Mottle Virus.

  Other internal and external pharmaceutical uses of the antimicrobial agents described herein include, but are not limited to, bacterial infections, tuberculosis, fungal infections such as yeast and mold infections (eg, Candida (eg, Candida Treatment of genus (eg Candida albicans, Candida glabrata, C. parapsilosis, C. tropicalis, and C. dubriniensis) or Cryptococcus or other fungi), Helicobacter pylori infection, and peptic ulcer disease or In one embodiment, the agent is at a dosage that is generally not lethal to bacteria but is still sufficient to reduce a protective polysaccharide coating that otherwise resists the natural immune response. Thus, this technology is used for human symbiotic microorganisms (eg normal gut microbiota) To help eradicate bacterial infections through the immune system to the extent that they can become cases with antibiotics without damage, by way of example and not limitation, certain contemplated embodiments are described herein. .

  Particulate bismuth-thiol for coating and treating water pipes In one embodiment, biofilm development is prevented and / or controlled (ie slowed, delayed, inhibited), biofilm disrupted, Or water pipes (eg, water pipes used by dentists, dental hygienists and other oral care professionals and therapists), or other water delivery vehicles such as pipes, pipes, faucets, fountains, showerheads, or humans Or any other instrument or device that contacts or delivers water consumed by or applied to non-human animals (eg, dental instruments, such as high-speed dental drills, air-water syringes, and The inner surface of a cleaning device or instrument (eg Cavitron®) or Provided herein is a method for reducing the amount of biofilm on the outer surface. These methods may also be useful to prevent, reduce, inhibit, eliminate or inhibit the growth and division of bacteria, fungi and / or protozoa in water pipes or water delivery vehicles. These methods include applying, flushing, adhering, or adhering the particulate BT compound to the surface of a water pipe or water delivery vehicle.

  A biofilm is a microscopic community consisting primarily of natural bacteria and fungi. Microorganisms form a thin layer on surfaces including dental water delivery systems and other water delivery vehicles such as showerheads, faucets and tubes. Water used as a cooling and cleaning agent in dental procedures can be severely contaminated by microorganisms (see, eg, Environmental Protection Agency website: epa.gov/safewater/mcl/html). Pathogenic microorganisms or opportunistic pathogens found in water from dental water pipes and instruments include Actinomyces, Bacteroides, Bacillus, Cropospodium, Escherichia, Flavobacterium, Klebsiella, Legionella Genus, Moraxella, Mycobacterium, Peptostreptococcus, Pseudomonas, Staphylococcus, Streptococcus, and Bayonella. Further, as a result of biofilm formation, Legionella species and protozoa can grow in water tubes or water delivery vehicles. Bacteria from the biofilm and other microorganisms present in the water pipe or water delivery vehicle are continuously released as a water stream through the water pipe or vehicle. Patients and clinical staff are exposed to microorganisms present in microdroplets or micromists sprayed from water pipes or water delivery vehicles.

  Regarding the use and consumption of water in dental applications, the US Centers for Disease Control and Prevention has determined that the number of bacteria in water used as a coolant / cleaner for non-surgical dental procedures is the number of aerobic heterotrophic plates (HPC) Is recommended to be ≦ 500 CFU / ml. The American Dental Association (ADA) advocates a more stringent standard and recommends that the water used for dental procedures contains bacterial levels of ≦ 200 CFU / ml. Measurements made to maintain low levels of bacteria in dental water systems include the use of antibacterial agents (see, eg, McDowell et al., J. Am. Dent. Assoc. 135: 799-805 (2004)). Hydrogen peroxide-based disinfectants (see, for example, Linger et al., J. Am. Dent. Assoc. 132: 1287-91 (2001)); conventional flushing of water pipes before and after use; Retention of delivery system; use of filtration system; use of chemicals as disinfectant (eg 1:10 diluted bleach, glutaraldehyde, food grade ethyl alcohol, chlorhexidine based product); thermal eradication; copper-silver ionization; Chlorine dioxide; UV; ozone; disinfectant combinations (eg, Adec® ICX Adex, Newburg, OR)); sodium percarbonate; silver nitrate and cationic surfactant and silver ion catalyst.

  Prevent and / or control biofilm development (ie slow, slow, inhibit), disrupt biofilm, or reduce the amount of biofilm on the inner or outer surface of a water pipe or water delivery vehicle Alternative antimicrobial agents that can be used to reduce include particulate BT compounds (or compositions comprising at least one particulate BT compound). The particulate BT compound can be introduced into water pipes, water pipe systems and water delivery vehicles, either manually or automatically, as gels, sprays, pastes, liquids or powders or other forms known to those skilled in the art. In certain embodiments, the particulate BT compound is mixed with at least one additional ingredient, either in powder or liquid form, which includes at least one additional biologically active ingredient and / or biological The product is formulated with an active excipient and delivered or injected periodically into a water pipe, water delivery vehicle or water system. The composition can be prepared by one skilled in the art using any number of methods known in the art. As an example, an antimicrobially effective amount of particulate BT compound can be used in combination with DMSO. For routine use, a level of particulate BT compound that is sufficient to prevent biofilm formation is desired. However, in other embodiments, the level of particulate BT compound is higher to reduce, remove, disrupt, or eliminate biofilms present in water pipes, water delivery vehicles, or water pipe systems. There is.

  The particulate BT compound can also be formulated to be gradually released from a composition comprising the particulate BT compound applied to a water pipe, water delivery vehicle or water pipe system. The particulate BT compound can also be incorporated into the coating, but it can be applied to, fixed, glued, or in some way contacted to the inner surface of a water pipe, vehicle or system. . The composition comprising the particulate BT compound can be a gel (eg, hydrogel, thiomer, airgel or organogel) or liquid. The organogel can comprise an organic solvent, lipoic acid, vegetable oil or mineral oil. The slow release composition may be applied with an antibacterial effective amount of the particulate BT compound for 1, 2, 3, 4, 5, 6 or 7 days (1 week), or 2, 3, 4, 5, 6, 7 weeks, or Delivery can be for 1, 2, 3, 4, 5 or 6 months.

  The particulate BT compound (or a composition comprising the particulate BT compound), when administered in combination, has at least one or more other having enhanced or synergistic antibacterial activity as described herein. It can be combined with antibacterial agents (ie, second, third, fourth, etc.). As an example, an enhanced antimicrobial effect can be observed when the particulate BT compound is administered together with an antimicrobial agent that chelates iron. The particulate BT compound can be combined with at least one of an oxidizing agent, a microbicide or a disinfectant. Particulate BT compounds prepared with hydrophobic thiols (eg thiochlorophenol) are used, which have a greater ability to adhere to water pipes and surfaces of water delivery vehicles and systems than low hydrophobic BT compounds. Can show. BT compounds having a net negative charge, such as those having a 1: 2 molar ratio (bismuth to thiol) may also have favorable adhesive properties.

  The particulate BT compound (and the composition comprising the particulate BT compound) can be combined with baking soda or another alkaline compound or substance. Due to the chemical and physical properties of baking soda, it has a wide range of uses such as cleaning, deodorizing and buffering. Baking soda is chemically neutralized rather than masking or absorbing odors. Baking soda may be combined with the particulate BT compound as a mixture of powders or dissolved or suspended in the powders, sprays, gels, pastes or liquids described herein. In other embodiments, the particulate BT compound can be used to maintain a desired alkaline pH and with other alkali metal bicarbonate or carbonate materials (eg, potassium bicarbonate or calcium carbonate) that also possess cleansing and deodorizing properties. Can be combined.

  As an additional example, the particulate BT compound (or a composition comprising the particulate BT compound) can be combined with one or more of the following. Antibacterial agents: eg chlorhexidine; sanguinarine extract; metronidazole; quaternary ammonium compounds (eg cetylpyridinium chloride); bis-guanides (eg chlorhexidine digluconate, hexetidine, octenidine, alexidine); halogenated bisphenol compounds (eg 2,2'methylenebis- (4-chloro-6-bromophenol) or other phenolic antibacterial compounds); alkyl hydroxybenzoates; cationic antimicrobial peptides; aminoglycosides; quinolones; lincosamides; penicillins: cephalosporins; Tetracycline; other antibiotics known in the art; Coleus forskori essential oil; silver or silver colloid antimicrobial agent; tin or copper based antimicrobial agent; Le; oregano; time; rosemary; or other herbal extracts; and grapefruit seed extract. Caries prevention agents: for example, sodium fluoride and stannous fluoride, amine fluoride, sodium monofluorophosphate, sodium trimetaphosphate, zinc citrate or other zinc agents, and casein. Plaque buffer: for example, urea, calcium lactate, calcium glycerophosphate, and strontium polyacrylate. Vitamins: For example, vitamins A, C and E. Plant extract. Anticalculus agents: for example, alkali metal pyrophosphates, hypophosphite-containing polymers, organic phosphonates and phosphocitrates. Biomolecule: For example, bacteriocin. Preservative. Opacifier. pH adjuster. Sweetener. Surfactants: For example, anionic, nonionic, cationic and zwitterionic or amphoteric surfactants, saponins from plant materials (see, eg, US Pat. No. 6,485,711). Particle abrasive: for example, silica, alumina, calcium carbonate, dicalcium phosphate, calcium pyrophosphate, hydroxyapatite, trimetaphosphate, insoluble hexametaphosphate, agglomerated particle abrasive, chalk, finely pulverized natural chalk, and the like. Moisturizer: for example, glycerol, sorbitol, propylene glycol, xylitol, lactitol and the like. Binders and thickeners: eg sodium carboxymethylcellulose, hydroxyethylcellulose (Natrosol®), xanthan gum, gum arabic, synthetic polymers (eg polyacrylates and carboxyvinyl polymers such as Carbopol®). Polymeric compounds that enhance the delivery of active ingredients such as antimicrobial agents. Buffers and salts for buffering the pH and ionic strength of the oral care composition. Bleach: For example, peroxy compounds (eg, potassium peroxydiphosphate). Foaming system: for example, sodium bicarbonate / citric acid system.

  In another embodiment, the particulate BT compound described herein (or a composition comprising a particulate BT compound) controls biofilm generation, disrupts biofilm, or reduces the amount of biofilm. Thus, it can be combined with at least one or more anti-biofilm agents. As understood in the art, interspecies quorum sensing is associated with biofilm formation. Certain agents that increase LuxS-dependent pathways or interspecies quorum sensing signals (see, eg, US Pat. No. 7,427,408) contribute to controlling biofilm development and / or growth. Exemplary agents include, by way of example, N- (3-oxododecanoyl) -L-homoserine lactone (OdDHL) blocking compounds and N-butyryl-L-homoserine lactone (BHL) analogs in combination or separately. (See, eg, US Pat. No. 6,455,031). An oral hygiene composition comprising a particulate BT compound and at least one anti-biofilm agent can be locally delivered for the disruption and inhibition of bacterial biofilms and for the treatment of periodontal disease (eg, For example, see US Pat. No. 6,726,898).

  The effectiveness of the particulate BT compound as an anti-biofilm agent heats the water pipe, water delivery vehicle or water pipe system to which the particulate BT compound is applied by heating the water pipe, water delivery vehicle or water pipe system. Can be enhanced. In certain embodiments, the water pipe, water delivery vehicle or water pipe system is heated to about 37 ° C. to about 60 ° C. or about 37 ° C. to about 100 ° C. In other embodiments, the water pipe, water delivery vehicle or water pipe system is about 45 ° C to about 50 ° C; about 50 ° C to about 55 ° C; about 55 ° C to about 60 ° C; about 60 ° C to about 70 ° C; About 70 ° C to about 80 ° C; about 80 ° C to about 90 ° C; or about 90 ° C to about 100 ° C. In certain embodiments, the water pipe, water delivery vehicle or water pipe system is heated to about 37 ° C. In another embodiment, the water pipe, water delivery vehicle or water pipe system is heated to 55 ° C. As will be appreciated by those skilled in the art, the length of time that the water pipe, water delivery vehicle or water system is heated will depend on the temperature applied. For example, the length of time required to achieve the same antibacterial effect is that the water pipe, water delivery vehicle or water system is heated to a lower temperature than is required when heated to a higher temperature. The case is longer. Determination of the appropriate length of time for exposure of the water pipe, water delivery vehicle or water pipe system at each temperature can be readily determined by one skilled in the art.

  The particulate BT compound (or a composition comprising the particulate BT compound) can be used in conjunction with other therapies to reduce or prevent biofilm development. By way of example, the particulate BT compound can be combined with an oxidizing chemical, tartar removal compound, biofilm disintegrant or flushing system as described herein and used in the art.

  Compositions comprising particulate bismuth-thiol and use for dental restorations In another embodiment, compositions comprising particulate BT compounds and dental amalgam and particulate BT compounds and dental for use in preventing and / or treating caries Composite materials are provided herein. In general, the only treatment for carious lesions is dental restoration by placement of an inert substance that acts as a barrier to further caries. Dental amalgams and dental composites are most commonly used for the restoration of teeth affected by cavity.

  Recurrent peripheral caries is an important contributor to repair failure, especially when dental composites are used for repair. The presence of bacteria inhabiting the interface between the composite material and the dental tissue can be an important factor in repair failure (see, eg, Hansel et al, J. Dent. Res. 77: 60-67 (1998)). ). A study in Portugal (Casa Pia Study, 1986-1989) confirmed 1,748 posterior repairs, of which 177 (10.1%) failed during the study. Recurrent peripheral caries accounted for 66% (32/49) and 88% (113/129) failure rates, respectively, due to the main reasons for failure in amalgam and composite repair (Bernardo et al. JADA 2007; 138: 775-83). Polymerization shrinkage, the shrinkage that occurs during the composite curing process, is closely linked as the main reason for postoperative peripheral leakage (eg Estefan et al., Gen. Dent. 2003; 51: 506-509). reference).

  Attempts have been made to mix antimicrobial compounds and agents into restorative materials such as dentin binding systems (DBS), but limited success has been achieved. The development of composite materials and amalgams and other restorative materials with antimicrobial properties can contribute to the prevention of secondary caries (see, for example, Imazato, Dent. Materials 19: 449 (2003)). This embodiment replaces the antimicrobial agent formulated using the repair composition described herein described in the art with the particulate BT compound described in the present invention. Provides the benefits disclosed in, e.g., antimicrobial activity, solubility and bioavailability, anti-biofilm action, non-toxicity, enhanced antibiotic efficacy, and other properties as described herein I intend to.

  In certain embodiments, a composition comprising a particulate BT compound and a dental composite material is provided. Dental composites typically contain a polymerizable resin base containing a ceramic filler. The particulate BT compound can be combined with any of the dental composite materials known in the art using methods practiced in the art (eg, Chicago: Quintessence Publishing Co.) (2002); et al. , Dental Materials: Properties and Manipulation (New York: Mosby) (2007); Roeters et al. , J. et al. Dent. 32: 371-77 (1998)).

In other embodiments, a composition comprising a particulate BT compound and an amalgam is provided. Amalgam is an alloy of mercury and one or more other metals. Most dental amalgams are called silver amalgams because silver is the main constituent that reacts with mercury. The kinetics of the reaction between mercury and silver is not suitable for clinical use, so silver is provided as an alloy with other elements. This alloy is often referred to as a dental amalgam alloy, or collectively, the alloys are known as “alloys for dental amalgam” (eg, International Standards Standard ISO 1559, Dental Materials-Alloys for (See Dental Amalgam (1995)). Several types of dental amalgam alloys are known, all containing tin, most having some copper, and a little zinc. Some of the dental amalgam alloys themselves contain a small amount of mercury to facilitate the amalgamation reaction. Conventional dental amalgam alloys contain 67% to 74% silver, 25 to 28% tin, and up to 6% copper and 2% zinc and 3% mercury. The so-called dispersed amalgam alloy has about 70% silver, 16% tin and 13% copper. A different group of amalgam alloys may contain up to 30% copper, which is known as a high copper content amalgam alloy. The amalgam alloy is mixed with mercury prior to clinical placement in a 1: 1 weight ratio. The mercury content of the finished dental amalgam restoration is therefore about 50% by weight. In conventional dental amalgam alloys, the silver to tin ratio results in a crystal structure that is essentially an intermetallic compound Ag3Sn called the gamma (γ) phase. The exact percentage of this phase controls the kinetics of the amalgamation reaction, as well as the numerous properties of the resulting amalgam structure. In high copper dispersion alloys, the microstructure is usually a mixture of gamma phase and eutectic silver-copper phase. Different manufacturers have different amalgam alloy formats, but they are usually spherical or irregular, available as fine particles with a particle size of about 25-35 microns (Scientific Committed and Newly Identified Health). Risks (SCENHR), European Commission: Directorate-General, Health & Consumer Protection, May 6, 2008 at Internet site:
ec. europa. eu / health / ph_risk / commites / 04_scenhr / docs / scenhr_o_016. pdf. reference).

  The particulate BT compound prevents or treats caries and / or inflammation by administering the particulate BT compound to the surface of the tooth, amalgam or composite (ie, the likelihood of occurrence or recurrence of caries and / or inflammation, respectively) Can also be used. The composition comprising the particulate BT compound can be a mucoadhesive composition applied to the surface of teeth and / or gingiva or oral mucosa, adheres to the surface to some extent, or a pharmaceutically effective amount of the desired surface. It can be in any form that delivers the active ingredient (s). The particulate BT compound can also be formulated to release gradually from the composition applied to the teeth. For example, the composite material can be a gel (eg, hydrogel, thiomer, airgel or organogel) or liquid. The organogel can comprise an organic solvent, lipoic acid, vegetable oil or mineral oil. Such gel or liquid coating formulations can be applied inside or outside an amalgam or composite or other restorative composition. The slow release composition comprises a pharmaceutically effective amount of the particulate BT compound for 1, 2, 3, 4, 5, 6 or 7 days (1 week), or 2, 3, 4, 5, 6, 7 weeks, or It can be delivered for 1, 2, 3, 4, 5 or 6 months. Such compositions can be prepared by one skilled in the art using any number of methods known in the art.

  Compositions containing particulate BT compounds that are useful for dental restorations include glass ionomer cements; gyomers (generated by reacting fluoride-containing glass and liquid polyacids); compomers (polymerizable dimethacrylate resins and ions Filterable glass-filled particles). The compomer can further comprise fluoride.

  The composition comprising the particulate BT compound applied to the surface of the tooth, amalgam or composite material may further comprise one or more other surfactants that enhance antimicrobial activity. Exemplary antimicrobial agents for use in compositions comprising particulate BT compounds include, for example, chlorhexidine, sanguinarine extract, metronidazole, quaternary ammonium compounds (eg, cetylpyridinium chloride); bisguanides (eg, chlorhexidine diglucose). Narate, hexetidine, octenidine, alexidine); and halogenated bisphenol compounds (eg, 2,2'-methylenebis- (4-chloro-6-bromophenol) or other phenolic antibacterial compounds; alkyl hydroxybenzoates; cations Aminoglycoside; quinolone; lincosamide; penicillin: cephalosporin; macrolide; tetracycline and other antibiotics; taurolidine or taurultam, A-dec ICX, Coleus forskori essential oil; silver or silver colloid antimicrobial agent; tin or copper based antimicrobial agent; chlorine or bromine oxidizing agent, manuka oil; oregano; thyme; rosemary; or other herbal extract; And grapefruit seed extract; anti-inflammatory or antioxidants such as ibuprofen, flurbiprofen, aspirin, indomethacin, aloe vera, turmeric, olive leaf extract, clove, panthenol, retinol, omega-3 fatty acid, gamma linolene Acid (GLA), green tea, ginger, grape seed and the like can be mentioned.

  The composition may further include one or more pharmaceutically acceptable carriers, such as starch, sucrose, water or water / alcohol systems, DMSO, and the like. The composition may also include a surfactant, such as an anionic, nonionic, cationic and zwitterionic or amphoteric surfactant, or may include a saponin derived from plant material (eg, US Pat. 6,485,711). Buffers and salts may also be included to buffer the pH and ionic strength of the composition for oral use. Other optional ingredients that may be included are bleaching agents such as peroxy compounds; potassium peroxydiphosphate; foaming systems such as sodium bicarbonate / citric acid systems and the like.

Composition comprising particulate bismuth-thiol and use for oral hygiene and treating oral inflammation and infection In another embodiment, a composition comprising particulate BT compound is formulated for oral use May be used in methods for preventing or reducing microbial growth of and for preventing and / or treating microbial infection and inflammation of the oral cavity. Therefore, these compositions are used to prevent or treat plaque, bad breath, periodontal disease, gingivitis, and other infections of the mouth (ie, reduce or inhibit their onset, and develop or reoccurrence). Useful for reducing likelihood). An oral composition comprising a particulate BT compound prevents and / or controls the development of biofilm (ie slows, delays, inhibits), disrupts biofilm, or the oral surface, in particular teeth or It can also be useful to reduce the amount of biofilm present on the gums.

  Entrained food particles, poor oral hygiene and poor oral health, and inadequate cleaning of complete dentures can promote microbial growth on teeth, gum periphery, and tongue. The continued growth of microorganisms and the presence of caries can lead to bad breath, plaque (ie, a biofilm formed by microbial colonization), gingivitis, and inflammation. If proper oral care (eg, brushing, flossing) is not performed, more severe infections such as periodontal disease and jaw infection can result.

  Good oral hygiene is important not only for oral health, but also for the prevention of multiple chronic conditions. Controlling oral bacterial growth can help reduce the risk of heart disease, maintain memory, and reduce the risk of infection and inflammation in other areas of the body. People with diabetes have a high risk of developing severe gum problems, and reducing the risk of gingivitis by maintaining good oral health can help control blood sugar levels. Pregnant women are more likely to suffer from gingivitis, and several studies have suggested an association between gum disease in pregnant women and the delivery of preterm and low birth weight infants.

  Bacteria are a major etiological agent in periodontal disease. More than 500 strains may be found in plaque (Kroes et al., Proc. Natl. Acad. Sci. USA 96: 14547-52 (1999)). Bacteria have evolved to survive as biofilms on the tooth surface, gingival epithelium, and oral environment, which makes it difficult to treat periodontitis. Bactericides and antibiotics currently used to treat such infections often do not kill all of the problematic organisms. Use of a drug that is ineffective against a particular strain may result in the growth of resistant strains. In addition, these agents can induce undesirable side effects, such allergic reactions, inflammation, and tooth discoloration.

  Dental bacterial plaque is a biofilm that adheres stubbornly to tooth surfaces, restorations, and prostheses. The main means of controlling the biofilm in the mouth is by mechanical cleaning (ie tooth brushing, flossing, etc.). In the absence of such cleaning, within 2 days thereafter, the tooth surface is mainly colored by facultative Gram-positive cocci, which are mainly Streptococcus. Bacteria secrete an extracellular mucus layer that helps fix bacteria to the surface and protect the attached bacteria. When the tooth surface is covered with attached bacteria, microcolony formation begins. Biofilms grow primarily through adherent bacterial cell division rather than the attachment of new bacteria. The doubling time of plaque formed by bacteria is rapid during early development and is slower in more mature biofilms.

  Co-aggregation occurs when engraftment continues to adhere to bacteria already attached to the membrane. As a result of co-aggregation, a complex array of different bacteria connected to each other is formed. A few days after plaque formation without interruption, the gingival margin becomes inflamed and swollen. Inflammation may form deep gingival crevice. The biofilm expands into this subgingival area and grows in this protected environment, forming a mature subgingival plaque biofilm. Gingival inflammation is not observed until the biofilm is changed from one composed mostly of gram positive bacteria to one containing gram negative anaerobic bacteria. Subgingival bacterial microcolonies composed mainly of gram-negative anaerobic bacteria become established in the gingival crevice between 3-12 weeks after plaque formation on the gingival margin begins. Currently, most bacterial species suspected of periodontal pathogens are anaerobic gram-negative bacteria.

  Bacterial microcolonies protected within the biofilm are typically resistant to antibiotics (systemic administration), disinfectants or bactericides (local administration), and immune defenses. For example, the antibiotic dose that kills airborne bacteria needs to be increased by 1500 times the dose that kills biofilm bacteria. At this high concentration, these antimicrobial agents tend to be toxic to patients (see, eg, Coghlan 1996, New Scientist 2045: 32-6; Elder et al., 1995, Eye 9: 102-9). .

  Careful and frequent physical removal of bacterial plaque biofilm is the most effective means of eliminating and controlling plaque. However, subgingival plaque in the pocket cannot be reached by brush, floss, or mouth rinse. Therefore, frequent periodontal debridement of the subgingival root surface by a dental hygienist or dentist is an essential element in the prevention and treatment of periodontitis.

  In certain embodiments, the particulate BT compound is an oral hygiene composition that can be routinely used by any subject, such as toothpaste, mouthwash (ie, mouth rinse), oral gel, toothpaste, oral spray (oral inhalation). Etc.), edible films, chewing gum, oral slurries, complete denture cleaning solutions, complete denture preservatives, dental floss, etc., for example, on coatings or in equipment. Particulate BT compounds are used in oral hygiene compositions used primarily by dental care professionals, including, for example, liquid fluorine treatments, cleaning compositions, buffing compositions, oral rinses, dental floss and cleaning tools, and devices. May be incorporated above. Embodiments of the present invention replace the antimicrobial agent formulated with the oral hygiene compositions described in the art and / or coated on the device with the particulate BT compounds described herein, Disclosed herein, including antibacterial activity, solubility and bioavailability range, anti-biofilm effects, non-toxicity, enhanced antibiotic efficacy, and other properties described herein Contemplates providing benefits.

  The particulate BT compound is used to prevent or treat caries and / or inflammation by administering the particulate BT compound to the tooth surface (ie, to reduce the likelihood of occurrence or recurrence of caries and / or inflammation, respectively). May be used). The composition comprising the particulate BT compound may be a mucoadhesive composition applied to the surface of the teeth and / or gums or oral mucosa, any form that adheres to the surface to some extent, or a pharmaceutically effective amount. In any form that delivers the active ingredient to the desired surface. The particulate BT compound can also be formulated so as to be slowly released from the composition applied to the teeth. For example, the composition may be a gel (eg, a hydrogel, thiomer, aerogel, or organogel) or a liquid. The organogel may include an organic solvent, lipoic acid, vegetable oil, or mineral oil. Such gel or liquid coating formulations may be applied to the inside or outside of an amalgam or composite or other restorative composition. The sustained release composition provides a pharmaceutically effective amount of the particulate BT compound for 1, 2, 3, 4, 5, 6, or 7 days (1 week), or 2, 3, 4, 5, 6, 7 It may be delivered weekly or for 1, 2, 3, 4, 5, or 6 months. Such compositions can be prepared by one skilled in the art using any number of methods known in the art.

  In other specific embodiments, as described herein, an antimicrobial composition comprising a particulate BT compound and one or more additional antimicrobial compounds or agents is provided for oral use. Particularly useful are compositions comprising a second antimicrobial agent that when administered in combination have a potent or synergistic antimicrobial effect as described herein. As an example, an enhanced antimicrobial effect may be observed when a particulate BT compound is administered with an antimicrobial agent that chelates iron. In other specific embodiments, the particulate BT compound is formulated with an anti-inflammatory agent, compound, small molecule, or macromolecule (eg, a peptide or polypeptide).

  Any of the particulate BT compounds described herein may be formulated for oral use. In certain embodiments, a particulate BT compound prepared with a hydrophobic thiol (eg, thiochlorophenol) may be used, which has the ability to adhere to tooth and mouth tissues more than a less hydrophobic BT compound. May be shown greatly. BT compounds having a net negative charge, such as those having a molar ratio of 1: 2 (bismuth to thiol), may have suitable adhesion.

  The oral hygiene composition comprising the particulate BT compound may further comprise one or more active ingredients and / or one or more orally suitable excipients or carriers. In one embodiment, the oral hygiene composition may further comprise baking soda or other alkaline compound or substance. Due to the chemical and physical properties of baking soda, it has a wide range of applications including washing, deodorizing, and buffering. Baking soda chemically neutralizes rather than masks or absorbs odors. Baking soda can be combined with the particulate BT compound as a powder mixture, or can be dissolved or suspended in any one of the toothpastes, gels, pastes, and liquids described herein. In other embodiments, the particulate BT compound is combined with other alkali metal bicarbonate or carbonate materials (eg, potassium bicarbonate or calcium carbonate) that help maintain the desired alkaline pH and also have cleaning and deodorizing properties. be able to.

The oral hygiene composition containing the particulate BT compound may further contain one or more of the following components.
Antimicrobial agents: eg chlorhexidine: sanguinarine extract, metronidazole, quaternary ammonium compounds (eg cetylpyridinium chloride); bisguanides (eg chlorhexidine digluconate, hexetidine, octenidine, alexidine); halogenated bisphenol compounds (eg 2,2'-methylenebis- (4-chloro-6-bromophenol) or other phenolic antibacterial compounds; alkyl hydroxybenzoates; cationic antimicrobial peptides; aminoglycosides; quinolones; lincosamides; Tetracycline; other antibiotics known in the art; Coleus forskori essential oil; silver or colloidal antimicrobial agent; antimicrobial agent based on tin or copper; Nukaoiru; oregano; time; rosemary; or other herbal extracts; and grapefruit seed extract.
Anti-inflammatory or antioxidant: for example, ibuprofen, flurbiprofen, aspirin, indomethacin, aloe vera, turmeric, olive leaf extract, clove, panthenol, retinol, ω-3 fatty acid, γ-linolenic acid (GLA), green tea , Ginger, grape seed, etc. Caries prevention agents: for example, sodium fluoride and stannous fluoride, amine fluoride, sodium monofluorophosphate, sodium trimetaphosphate, zinc citrate or other zinc agents, and casein. Plaque buffer: for example, urea, calcium lactate, calcium glycerophosphate, and strontium polyacrylate. Vitamins: For example, vitamins A, C and E. Plant extract. Desensitisers: For example, potassium citrate, potassium chloride, potassium tartrate, potassium bicarbonate, potassium oxalate, potassium nitrate, and strontium salts. Anticalculus agents: for example, alkali metal pyrophosphates, hypophosphite-containing polymers, organic phosphonates and phosphocitrates. Biomolecule: For example, bacteriocin, bacteriophage, antibody, enzyme and the like. Flavoring agents: for example, peppermint and spearmint oil, fennel, cinnamon. Proteinaceous material: for example, collagen. Preservative. Opacifier. Coloring agent. pH adjuster. Sweetener. Pharmaceutically acceptable carrier: for example starch, sucrose, water or water / alcohol system. Surfactants: For example, anionic, nonionic, cationic and zwitterionic or amphoteric surfactants, saponins from plant materials (see, eg, US Pat. No. 6,485,711). Particle abrasive: for example, silica, alumina, calcium carbonate, dicalcium phosphate, calcium pyrophosphate, hydroxyapatite, trimetaphosphate, insoluble hexametaphosphate, agglomerated particle abrasive, chalk, finely pulverized natural chalk and the like. Moisturizer: for example, glycerol, sorbitol, propylene glycol, xylitol, lactitol and the like. Binders and thickeners: for example sodium carboxymethylcellulose, hydroxyethylcellulose (Natrosol®), xanthan gum, gum arabic, synthetic polymers (for example polyacrylates and carboxyvinyl polymers such as Carbopol®). Polymeric compounds that improve the delivery of active ingredients such as antimicrobial agents. Buffers and salts that buffer the pH and ionic strength of the oral care composition. Bleach: For example, peroxy compounds (eg, potassium peroxydiphosphate). Foaming system: for example sodium bicarbonate / citric acid system. Discoloration system. In certain embodiments, the abrasive is silica or finely ground natural chalk.

  An oral hygiene composition comprising a particulate BT compound formulated for use as a toothpaste further comprises a hygroscopic agent (eg, glycerol or sorbitol), a surfactant, a binder, and / or a flavoring agent. Also good. Toothpaste may contain sweeteners, whitening agents, preservatives, and antimicrobial agents. The pH of the composition for toothpaste and other oral uses is typically between pH 5.5 and 8.5. In certain embodiments, the oral hygiene composition, including toothpaste, has a pH of 7-7.5, 7.5-8, 8-8.5, or 8.5-9, and the particulate BT The antimicrobial activity of the compound can be enhanced. The toothpaste compositions described herein include chalk, dicalcium phosphate dihydrate, sorbitol, water, hydrated aluminum oxide, precipitated silica, sodium lauryl sulfate, sodium carboxymethylcellulose, flavoring agent, monoolein Acid sorbitan, sodium saccharin, tetrasodium pyrophosphate, methyl paraben, propyl paraben. If desired, one or more colorants such as FD & C Blue can be used. Other suitable ingredients that may be included in toothpaste formulations are described in the art, for example, in US Pat. No. 5,560,517.

  In one particular embodiment, the oral hygiene composition is a mouse spray and includes a particulate BT compound, an alkaline buffer (eg, potassium bicarbonate), an alcohol, a sweetening ingredient, and a flavoring system. The flavoring system may have one or more of the following: flavoring agents, hygroscopic agents, surfactants, sweeteners, and colorants (see, eg, US Pat. No. 6,579,513). ). The surfactants described herein and known in the art for use in oral hygiene compositions may be anionic, nonionic or amphoteric.

  In another embodiment, particulate BT-containing oral hygiene compositions have been described in the art as being useful for inclusion in toothpastes, toothpaste gels, mouthwashes, and the like to treat severe infections. It may be combined with further active ingredients such as taurolidine and taurultam (see, for example, British Patent Application GB 1557163, US Pat. No. 6,488,912). As described herein, microparticles BT can also be combined with one or more additional antimicrobial agents, when combined with microparticles BT, the combination has an additive or synergistic effect.

  In yet another specific embodiment, the oral hygiene composition described herein is at least one for controlling biofilm generation, for disrupting biofilm, or for reducing the amount of biofilm. Further or more anti-biofilm agents may be included. As understood in the art, interspecies quorum sensing is involved in biofilm formation. Certain agents that increase the LuxS-dependent pathway or interspecies quorum sensing signal (see, eg, US Pat. No. 7,427,408) contribute to the control of biofilm development and / or proliferation. Exemplary agents include, for example, combined or separate N- (3-oxododecanoyl) -L-homoserine lactone (OdDHL) blocking compounds and N-butyryl-L-homoserine lactone (BHL) analogs. (See, eg, US Pat. No. 6,455,031). An oral hygiene composition comprising a particulate BT compound and at least one anti-biofilm agent can be delivered topically for disintegration and inhibition of bacterial biofilm and treatment of periodontal disease (eg, US, for example) No. 6,726,898).

  The oral hygiene compositions described herein may contain a sufficient amount of particulate BT compound to perform substantial antimicrobial action at the time required for normal tooth brushing, mouth rinsing, or flossing. Good. As described herein, the particulate BT compound may be retained on the oral surface (eg, teeth, amalgam, composites, mucous membranes, gums). For example, particulate BT compounds retained on teeth and gums after brushing, rinsing, and flossing may continue to provide long-term anti-biofilm and anti-inflammatory effects.

  In other embodiments, the particulate BT compound is slowly released from a mucoadhesive polymer or other agent that contributes to retention of the particulate BT compound on the mucosa, teeth and restoration surfaces. The particulate BT compound may be added to a stable viscous mucoadhesive aqueous composition for the prevention and treatment of mucosal ulcerous, inflammatory and / or erosive disorders and / or topical treatment. It may be used for delivery of pharmaceutically active compounds to the mucosal surface or transfer to the systemic circulation (see, eg, US Pat. No. 7,547,433).

  In another embodiment, the oral hygiene composition comprising the particulate BT compound further comprises olive oil that can enhance plaque removal. The use of olive oil in products intended for oral hygiene, such as toothpastes, mouthwashes, sprays, oral inhalers, or chewing gum, eliminates or reduces (decreases) bacterial plaque and / or Contributes to the elimination or reduction (reduction) of the number of bacteria present, thereby achieving a reduction in the incidence of dental diseases (eg, caries, periodontal disease) and bad breath (eg, US Pat. No. 7,074) 391).

  In other embodiments, the oral hygiene composition comprising the particulate BT compound may further comprise a mucosal bactericidal preparation for topical application in the mouth. The oral hygiene composition may further comprise an aqueous slurry useful for washing the tongue and throat (eg, US Pat. No. 6,861,049). In yet another embodiment, the oral hygiene composition comprising the particulate BT compound is at least used to prevent the formation of cavities (ie, reduce the likelihood of occurrence) or reduce the number of cavities. One mint may further be included. One such mint called CaviStat® (Ortek Therapeutics, Inc., Roslyn Heights, NY) contains arginine and calcium, helps neutralize acidic pH, and adheres to calcium on enamel surfaces Promote. Inclusion of mint in the oral hygiene composition containing the particulate BT compound can thus increase the pH and enhance the adhesion of the particulate BT compound to the oral surface.

  Adhesive composition comprising particulate bismuth-thiol formulated for dental and orthopedic use In another embodiment, the composition comprising particulate BT compound is on a bone or joint prosthesis (prosthesis), or Formulated for use in a method for preventing or reducing microbial growth of tissues and skeletal structures adjacent to bone or joint prosthetics (prosthetic joints). In certain embodiments, to prevent and / or prevent microbial infection and inflammation caused by orthopedic procedures (eg, orthopedic surgery, orthopedic treatment, arthroplasty (including two-stage arthroplasty), orthodontic treatment) A method of using a composition comprising a particulate BT compound for treatment is provided. In certain embodiments, the composition comprises a particulate BT compound and bone cement, and in certain other embodiments, the composition comprises a particulate BT compound and dental cement. Thus, these compositions prevent and / or treat (i.e. reduce or inhibit the onset of) microbial infections of the skeleton and support structures (i.e. bones, joints, muscles, ligaments, tendons), e.g. osteomyelitis, To reduce the likelihood of its occurrence or recurrence). Compositions described herein comprising a particulate BT compound and bone cement or dental cement prevent and / or control biofilm development (ie slow, retard, inhibit) biofilm disruption Also useful for reducing the amount of biofilm present in the joint, or on the surface of joints, bones, ligaments, tendons or teeth, or replacement joints, bones (partial or whole), ligaments, tendons or teeth It can be.

  The cements described herein and known in the art are binder materials that can bond materials together and cure. Such materials may bind the tissues together or may connect a replacement or prosthetic device (eg, a replacement joint, bone or tooth) to adjacent tissue. Examples of the bone cement include polymethyl methacrylate (PMMA), magnesium phosphate, and calcium phosphate. The calcium phosphate form is used as a “replacement bone” to treat bone cracks and cuts that cannot be healed sufficiently quickly and / or properly without the use of graft material. Compositions comprising bone cement (eg, calcium phosphate) and particulate BT compounds can also be used to treat cancellous bone defects by providing mechanical integrity with cancellous bone. The cement can be resorbed or retained at the site of implantation.

  In certain embodiments, a composition described herein that is useful as a bone cement comprises a BT compound or a particulate BT compound, and a preparation of calcium phosphate or magnesium phosphate suitable for use as a bone cement. . Preparations of calcium phosphate or magnesium sulfate may also be referred to herein as calcium phosphate containing bone cement or calcium phosphate bone cement, or magnesium phosphate containing bone cement or magnesium phosphate bone cement, respectively. Calcium phosphate is included in the composition in any one of several forms known and used in the art, including, but not limited to, hydroxyapatite (Ca10 (PO4) 6 (OH) 2) Brushite (CaHPO4 * 2H2O); monetite (CaHPO4); calcium deficient hydroxyapatite (CDHA, Cag (PO4) 5HPO4OH); calcium sulfate / phosphate (CSPC) (eg, Hu et al., J. Mater. Sci. Mater. Med. 2009 October 13, e-publication ahead of print). Magnesium phosphate used in the art is also referred to as struvite (MgNH4PO4 * 6H2O) cement (see, eg, Groshardt et al., Tissue Eng. Part A, 2010 Jul 30, e-pub ahead of print; see, eg, Boh al., J. Pharm. Sci. 86: 565-72; (1997); Fulmer et al., 3: 299-305 (1992); Lobenhoffer et al., J. Orthopaed Trauma 16: 143-49 (2002). Lee et al., J. Carniofac. Surg. 21: 1084-88 (2010)). In one particular embodiment, a composition described herein comprising a particulate BT compound and a calcium phosphate-containing bone cement comprises calcium sulfate / phosphate (CSPC) as a form of calcium phosphate (eg, Hu et al., J. Mater. Sci. Mater. Med. 2009 October 13, e-publication ahead of print). In certain other embodiments, the composition comprising the particulate BT compound and calcium phosphate or magnesium phosphate cement comprises chitosan (a biopolymer from crustacean cells); at least one or more antibiotic or antimicrobial agent; and / or at least One or more anti-inflammatory agents may further be included.

  Bone cement has been used in the art for the release of drugs and agents. In certain embodiments, the calcium phosphate cement may be at least partially in the form of hydroxyapatite microspheres encapsulating an agent (eg, an antimicrobial agent) for therapeutic use (eg, US Pat. No. 6, 730,324). Such cements containing microspheres are useful for the slow release of agents contained within the microspheres. Contemplated herein is a composition comprising calcium phosphate microspheres comprising a particulate BT compound.

  Compositions comprising a particulate BT compound and a PMMA bone cement are also provided herein. PMMA bone cement can be formulated with particulate BT compounds according to methods described in the art to formulate PMMA with other agents having antimicrobial activity (see, eg, European Patent Application EP 1649874).

  Also provided herein is a composition comprising a particulate BT compound and a dental cement (ie, a dental adhesive), the composition for inhibiting, preventing or treating microbial infections of teeth or gums. Can be used. The dental cement may include any one of the following compounds or compositions: zinc phosphate, glass ionomer, alpha-tricalcium phosphate (α-TCP), alkyl methacrylate (eg, US Pat. No. 6, No. 071,528); bismuth oxide (see, for example, Bueno et al., Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endod. 107: e65-69 (2009)); MTA) (see, eg, Hwang et al., Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endod. 107: e96-102 (2009)).

Embodiments of the present invention disclose herein, replacing the antimicrobial agents formulated with dental or bone cements described in the art with the particulate BT compounds described in the present invention. Providing benefits such as antimicrobial activity, solubility and bioavailability, anti-biofilm action, non-toxicity, enhanced antibiotic efficacy, and other properties as described herein Intended. Bone and dental cements can be formulated using particulate BT compounds and one or more additional components according to methods described in the art (see, eg, US Patent Application No. 2006/0205838; Alt et al. Agents Chemother. 48: 4-84-88 (2004); Bohner et al., Supra; Chuen et al., Antimicrob. J. Orthopaed.Res.27: 1008-15 (2009); De Lalla, J. Chemother.13: 48-53 (2001); Domenico et al., Peptides. 5:... 2047-53 (2004); Widmer et al, Antimicrob Agents Chemother 35: 741-46 (1991)).
reference).

  The amount of BT compound used in the particulate BT compound-containing composition comprising bone cement or dental cement can range from about 10 to 500 μg BT / g of each cement. The particulate BT compound, alone or in combination with at least one additional component, provides advantages as described herein over the antibiotics currently used in bone and dental cement. . The compositions described herein comprising a particulate BT compound and bone cement (eg, calcium phosphate) or dental cement may further comprise one or more additional antimicrobial compounds or antimicrobial agents. Particularly useful are compositions comprising a particulate BT compound and a second antimicrobial agent that, when combined, has an enhanced or synergistic antimicrobial action as described herein. As an additional example, enhanced antimicrobial activity can be observed when the particulate BT compound is administered together with an antimicrobial agent that chelates iron. In other specific embodiments, the particulate BT compound is formulated with an anti-inflammatory agent, compound, small molecule or macromolecular substance (eg, a peptide or polypeptide).

  A composition comprising a particulate BT compound and a bone cement as described herein is a metal product used to attach, stabilize, or fix fractures, fusions, osteotomy or replacement joints (eg, , Screws, plates, staples, pins and wires, etc.). A composition comprising a particulate BT compound and a dental cement as described herein comprises a dental pulp, a tooth cap, an inner layer, a tooth, or a dental filling or restorative composition in a tooth, etc. Can be used to coat. These compositions can be formulated into a coating that can be applied to adhere and adhere to the surface of bone and / or joint-related metal products, or placed in contact in several ways. In certain embodiments, the coating comprises a particulate BT compound and calcium phosphate or magnesium phosphate bone cement. The particulate BT compound and calcium phosphate or magnesium phosphate are formulated together for application to bone metal products according to methods practiced in the art. For example, a composition comprising a particulate BT compound and a bone cement (eg, calcium phosphate or magnesium phosphate bone cement) is a liquid, gel, paste or spray (eg, thermal spray including plasma spray) for application to metal products. It can be in form. The composition comprising the particulate BT compound and bone cement can be a gel (eg, hydrogel, thiomer, aerogel or organogel) or liquid. The organogel can comprise an organic solvent, lipoic acid, vegetable oil or mineral oil. The slow release composition may be applied with an antibacterial effective amount of the particulate BT compound for 1, 2, 3, 4, 5, 6 or 7 days (1 week), or 2, 3, 4, 5, 6, 7 weeks, or Delivery can be for 1, 2, 3, 4, 5 or 6 months. The release rate can be controlled, at least in part, by the porosity of the cement (see, eg, Bohner et al., Supra).

  The composition comprising the particulate BT compound and bone cement or dental cement, when administered in combination, at least one other antimicrobial agent having an enhanced or synergistic antimicrobial effect (ie, greater than an additive effect) (Ie, second, third, fourth, etc. antimicrobial agents). As an example, an enhanced antimicrobial effect can be observed when the particulate BT compound is administered together with an antimicrobial agent that chelates iron. In certain embodiments, the composition comprising the particulate BT compound and bone cement or dental cement may be combined with at least one other antibacterial and / or anti-inflammatory agent selected from: Chlorohexidine; sanguinarine extract; metronidazole; quaternary ammonium compounds (eg cetylpyridinium chloride); bis-guanides (eg chlorhexidine digluconate, hexetidine, octenidine, alexidine); halogenated bisphenol compounds (eg 2,2 ' Methylene bis- (4-chloro-6-bromophenol) or other phenolic antibacterial compounds); alkyl hydroxybenzoates; cationic antibacterial peptides; aminoglycosides; quinolones; lincosamides; Macrolides; Tetracycline; Other antibiotics known in the art; Coleus forskori essential oil; Silver or silver colloid antimicrobial agent; Antimicrobial agent based on tin or copper; Manuka oil; Oregano; Time; Mary; or other herbal extracts; and grapefruit seed extract. Anti-inflammatory or antioxidant: for example, ibuprofen, flurbiprofen, aspirin, indomethacin, aloe vera, turmeric, olive leaf extract, clove, panthenol, retinol, ω-3 fatty acid, γ-linolenic acid (GLA), green tea , Ginger, grape seed, etc. In certain embodiments, the composition comprising the particulate BT compound and bone cement or dental cement is an antibiotic selected from clindamycin, vancomycin, daptomycin, cefazolin, gentamicin, tobramycin, metronidazole, cefaclor, ciprofloxacin Or other antimicrobial agents, such as quaternary ammonium compounds (eg, benzalkonium chloride, cetylpyridinium chloride), antimicrobial zeolites, alkali metal hydroxides, or alkaline earth metal oxides . The composition optionally comprises one or more pharmaceutically suitable carriers (ie, excipients), surfactants, buffers, diluents, and salts, and bleaches described herein. May be included. Antimicrobial agents can be formulated together with dental and bone cements as described herein and in the art (eg, Akashi et al., Biomaterials 22: 2713-17 (2001); US patents). 6,071,528; Alt et al., See above).

  Animal models of foreign body infection can be used to characterize the antimicrobial activity of compositions comprising particulate BT compounds and death song or bone cement (eg, Chuard et al., Antimicrob. Agents Chemother. 1993; 37 : 625-32). The in vivo efficacy of antibiotics in these models correlates with the ability of antimicrobials to kill stationary microorganisms as well as those that are adherent to foreign substances (eg, Widmer et al. J. Infect. Dis. 1990; 162: 96-102; see Widmer et al. Antimicrob Agents Chemother 1991; 35: 741-6; see also, for example, Karchmer. Clin. Infect. Dis. 1998; 27: 714-6).

  As a non-sourced example and for illustrative purposes only, bone cement is composed of 75% (2/2) methyl methacrylate styrene copolymer, 15% polymethyl methacrylate (to help handle the composition) and 10% barium sulfate ( The particulate BT compound may be included in about 10 to about 500 μg of particulate BT compound / g of cement powder (ie, 0.001 to 0.05% w / w), as well as for radiopacity. In other specific embodiments, at least one additional antimicrobial agent may be added.

  Compositions comprising particulate bismuth-thiol formulated with paints and paint coatings Certain other embodiments reduce biofouling, prevent and / or control biofilm development (i.e. slow, slow, In order to disrupt the biofilm or reduce the amount of biofilm present on the painted surface, the particulate BT compounds described herein can be incorporated into the paint or as a paint coating on the paint. Intended to be incorporated. The compositions described herein comprising particulate BT compounds are numerous products such as medical devices, orthopedic devices, dental devices, industrial devices, electronic devices, walls, floors, ceilings, roofs. , Piles, wharfs, piers, conduits, pipelines and plumbing structures (eg intake screens, cooling towers), heat exchangers, dams and fabrics, and other surfaces such as cars, trains, airplanes and ships, eg large May be formulated with paint or paint coating applied to any one of (but not limited to) surfaces present in and on all types of vehicles including ships, boats, submarines and other ships .

  In one particular embodiment, the compositions and methods described herein are useful for preventing and / or reducing bioadhesives or biofilms formed on products exposed to water. is there. The formation of biofilms on the surface in the marine environment is believed to be an important contributor to colonization and mobilization of several adherent invertebrate communities on marine structures (eg, Siboni et al , FEMS Microbilol Lett 2007; 274: 24-9). Subsequent interactions between the macrobiota and these microbial coats can invert most of the invertebrates and algae (which are the hydrodynamic drags associated with sex deposits) within days or weeks. (See, eg, Schultz, Biofouling 2007; 23: 331-41). Old biofilms on the surface supported barnacle larvae attachment regardless of the type of support (see, eg, Hunga et al., J Exptl Marine Biol Ecol 2008; 361: 36-41). Microfilaments also have significantly greater adhesion strength at one or more developmental stages in the squirts Phallusia nigra, polychaete helminths (Hydroides elegans) and barnacles (Balanus amphitrite). (See, for example, Zardus et al., Biol Bull 2008; 214: 91-8). Biofilms can also enhance the attachment of mussels, Dresena polymorpha, to some artificial surfaces (see, eg, Kavouras & Maki. Inverteb Biol 2005; 122: 138-51), Profits and expenses, if not billions, have been generated for marine foods, power generation and manufacturing, and for water and wastewater treatment facilities, resulting in significant damage to the ecosystem where mussels are introduced Is causing.

  In seawater, brackish water, and freshwater environments, organisms gather, clench, attach, and multiply on underwater structures and ships. Such organisms include algae, fungi and other microorganisms, and aquatic animals such as ascidians, hydroworms, bivalves, bryozoans, polychaetes, sponges and barnacles. The presence of these organisms, known as “attachments” of the structure, for example, adds to the weight of the structure and / or interferes with its hydrodynamics, thereby reducing its operating efficiency and being corroded. It can be detrimental by increasing ease and breaking down or crushing the structure.

  Certain paints and coatings used to date to prevent or reduce biofouling and biofilm production inhibit the biofouling and biofilm formation, while the desired and beneficial flora and Contains toxic components that can be toxic to fauna. Exemplary biocides and chemical toxins include copper and copper-containing compounds (eg, cuprous oxide), mercury, arsenic, tributyltin oxide (TBT), organotin (ie, one or more attached carbon groups). Tin), hexio 2-part bisphenol-A-epichlorohydrin epoxy compound, bifunctional glycidyl ether epoxy resin, glycidyl ether epoxy, and barium metaborate epoxy.

  The particulate BT compounds described in the present invention provide a non-toxic alternative and have the advantages disclosed herein, such as a range of antibacterial activity, solubility and bioavailability, anti-biofilm action Providing enhanced antibiotic efficacy, as well as other properties as described herein. Particulate BT compounds are substituted in place of other antimicrobial agents in paints and paint coatings, and are used herein using methods known for producing paints and paint coatings containing biocides. Integration of the particulate BT compounds and methods described can be incorporated into these paints and paint coatings (eg, US Pat. Nos. 4,596,724; 4,410,642; 4,788,302). No. 5,470,586; No. 6,162,487; No. 5,384,176; US Patent Application Publication Nos. 2007/125703 and 2009/0197003; Gerhart et al., J. Chem. 14: 1905-17 (1988); Sears et al., J. Chem. 16: 791-99 (1990); Ganguli et al., Smart Mater. Struct.18: 104027 (2009); Cao et al., ACS Applied Materials Interface 1: 494 (2009); Materials 7: 236-41 (2008)). Examples of the coating material into which the fine particle BT compound can be mixed include a coating material based on epoxy, silicone, or acrylic. In a more specific embodiment, the particulate BT compound can be incorporated into paints formulated for marine use and exposure to seawater, examples of which include alkyd resin substrates, bitumen substrates, gilsonite Examples thereof include paints for base materials, chlorinated rubber base materials, and epoxy resin base materials.

  Antimicrobial agents can be released in a controlled manner by incorporating the agent into the paint coating. Methods for enhancing the rate of drug release from composite materials are known in the art. The composite material may comprise a natural or synthetic bioabsorbable polymer matrix and a drug particle phase dispersed therein (see, eg, US Pat. Nos. 7,419,681 and 5,028,664; US See also patent application 2009/0043388). As an example, the drug eluting paint coating composition may include at least one particulate BT compound dispersed in a modified biologically active binder.

  The particulate BT compound can also be formulated to be gradually released from a composition comprising the particulate BT compound applied to the painted surface. The particulate BT compound can also be incorporated into a coating (eg, an epoxy coating) that is applied to secure and adhere to the surface of the painted structure or product, or that can be placed in contact in several ways. The particulate BT compound can be gradually released from such a composition. The slow release composition comprising the particulate BT compound can be a gel (eg, hydrogel, thiomer, aerogel or organogel) or liquid. The organogel can comprise an organic solvent, lipoic acid, vegetable oil or mineral oil. The slow release composition may be applied with an antibacterial effective amount of the particulate BT compound for 1, 2, 3, 4, 5, 6 or 7 days (1 week), or 2, 3, 4, 5, 6, 7 weeks, or Delivery can be for 1, 2, 3, 4, 5 or 6 months.

  Other coatings used in the art and with which the particulate BT compounds described herein can be formulated together include polysaccharide matrices such as polysaccharide matrices that are reversibly crosslinked with polyvalent metal cations (eg, U.S. Patent Application No. 2009/0202610); titanium nanotubes; nanostructured surfaces; biocompatible dextran-coated nanoceria (with pH-dependent antioxidant properties); polysorbone block polymers; as well as other biodegradable coatings ( For example, see US Pat. No. 6,162,487). Other coatings contemplated herein formulate particulate BT compounds with anti-corrosion and anti-fouling anti-bacterial coatings used in industry, including, but not limited to, carnauba wax fluoropolymer, xylan (registered) Trademark), PTFE and molybdenum materials.

  The particulate BT compound concentration (by weight) in the paint or paint coating can vary, for example, from as low as about 0.001% to about 0.1%, depending on the intended use and desired properties of the paint or paint coating. . A particulate BT compound (or a composition comprising a particulate BT compound) incorporated in a paint or paint coating has an enhanced or synergistic antimicrobial effect as described herein when administered in combination. , Can be combined with at least one other antimicrobial agent (ie, second, third, fourth, etc. antimicrobial agent). By way of non-limiting example, antimicrobial agents that can be included in compositions containing particulate BT compounds include: chlorhexidine; sanguinarine extract; metronidazole; quaternary ammonium compounds (eg, cetylpyridinium chloride); bisguanides (eg, chlorhexidine di). Gluconate, hexetidine, octenidine, alexidine); halogenated bisphenol compounds (eg, 2,2'-methylenebis- (4-chloro-6-bromophenol) or other phenolic antibacterial compounds; alkyl hydroxybenzoates; cationic Antimicrobial peptides; aminoglycosides; quinolones; lincosamides; penicillins: cephalosporins; macrolides; tetracyclines; other antibiotics known in the art; , Silver colloidal antimicrobial agent, tin or copper based antimicrobial agent, manuka oil, oregano, thyme, rosemary, or other herbal extract, and grapefruit seed extract. , And the surfactants, diluents or carriers, buffers and / or bleaches described above and herein.

  Compositions comprising particulate bismuth-thiol formulated with concrete and cement compounds Certain other embodiments prevent and / or control (ie slow, retard, inhibit) biofilm development In industrial cement and in or on concrete, mortar and grout (including concrete, mortar and grout coatings) to reduce the amount of biofilm present on the concrete surface It is intended to incorporate the particulate BT compound described in the document. Microorganisms that grow on and in concrete structures reduce the useful life of the product and can have health hazards to animals and humans that are exposed to microorganisms present on the concrete surface (see, eg, Idachaba et al., Res. 19: 284-91 (2001); Idachaba et al., J. Hazard. Mater. 90: 279-95 (2002); Tazaki, Canadian Minalogist 30: 431-34 (1992)).

  As used herein, as well as in the art, cement refers to a dry powder material (typically limestone that may also contain additional materials) used to bond concrete aggregate. Exemplary cements described in the art are commonly referred to as Portland cement, blast furnace cement, masonry cement, lime scrap cement and calcium aluminate cement. Upon the addition of water and / or additives, the cement mixture is called concrete, especially when aggregate is added. Concrete is a composite material composed of aggregates (eg gravel and sand), cement and water. Cement used in construction is characterized as hydraulic or non-hydraulic. Hydraulic cements are typically used for finishing brick buildings in wet climates; for stone buildings in harbor buildings that come into contact with seawater.

  Compositions described herein comprising particulate BT compounds are used for concrete structures such as bridges, buildings, pipes, elevated highways, tunnels, garages, submarine oil mining platforms, wharfs, dam walls, water systems and pipelines. Can be used to coat, or be mixed with, cement used for floors, worktops, walkways, roadways, loading docks, skateboarding structures, and radioactive waste primary storage structures obtain. The particulate BT compounds described herein can be incorporated into cement as described in the art (see, eg, US Pat. No. 7,507,281). The alkalinity of the cement or concrete can also enhance the antimicrobial action of the particulate BT compound.

  Cement can also be degraded by oxidizing bacteria, such as Thiobacillus thiooxidans. As a non-limiting example by way of illustration, the bismuth thiol compound BisEDT (but not the particulate BT compound described here) retards the growth of sulfur-oxidizing bacteria in concrete used for waste and radioactive waste disposal systems. It has been shown. The effective antibacterial range of BisEDT in concrete has been shown to be 10-500 μg / g or 0.001-0.05%. Higher BisEDT levels interfered with concrete strength. Other compounds, such as BisPYR, may be useful to inhibit mold and algae deposits and biofilm development. Embodiments of the present invention replace the bismuth-thiol compounds and other antimicrobial agents with the particulate BT compounds described in the present invention, such as the advantages disclosed herein, such as antimicrobial activity, solubility and biological It is intended to provide bioavailability, anti-biofilm action, non-toxicity, enhanced antibiotic efficacy, and other properties as described herein.

  The particulate BT compound can be introduced to the concrete surface manually or automatically as a gel, spray, paste, liquid or powder or other forms known to those skilled in the art. In certain embodiments, the particulate BT compound is mixed with at least one additional ingredient, either in powder or liquid form, which includes at least one additional biologically active ingredient and / or biological Is the product formulated with active excipients and delivered periodically in or on the concrete structure (ie, on the surface of the exposed concrete structure, particularly the surface exposed to water)? Or injected. The composition can be prepared by one skilled in the art using any number of methods known in the art. As an example, an antimicrobially effective amount of particulate BT compound can be used in combination with DMSO (eg, 1 mg / ml particulate BT compound in DMSO). For routine use, a level of particulate BT compound that is sufficient to prevent biofilm formation is desired. However, in other embodiments, the level of particulate BT compound may be higher to reduce, remove, disrupt or eliminate biofilm present on the concrete surface.

  The particulate BT compound can also be formulated to gradually release from a composition comprising the particulate BT compound applied to the surface of the concrete structure. Particulate BT compounds can also be incorporated into coatings (eg, epoxy coatings), which are applied to the surface of a concrete structure, secured, glued or arranged in some way to contact. Can be done. The particulate BT compound can be gradually released from such a composition. The slow release composition comprising the particulate BT compound can be a gel (eg, hydrogel, thiomer, aerogel or organogel) or liquid. The organogel can comprise an organic solvent, lipoic acid, vegetable oil or mineral oil. The slow release composition may be applied with an antibacterial effective amount of the particulate BT compound for 1, 2, 3, 4, 5, 6 or 7 days (1 week), or 2, 3, 4, 5, 6, 7 weeks, or Delivery can be for 1, 2, 3, 4, 5 or 6 months.

  The particulate BT compound (or a composition comprising the particulate BT compound), when administered in combination, has at least one or more other having enhanced or synergistic antimicrobial effects as described herein. Can be combined with antimicrobial agents (ie, second, third, fourth, etc. antimicrobial agents). As an example, an enhanced or synergistic antimicrobial effect can be observed when the particulate BT compound is administered with an antimicrobial agent that chelates iron. The particulate BT compound can be combined with at least one other antimicrobial agent, including a fungicide or algicidal agent. By way of non-limiting example, antimicrobial agents that can be included in compositions containing particulate BT compounds include: chlorhexidine; sanguinarine extract; metronidazole; quaternary ammonium compounds (eg, cetylpyridinium chloride); bisguanides (eg, chlorhexidine di). Gluconate, hexetidine, octenidine, alexidine); halogenated bisphenol compounds (eg, 2,2'-methylenebis- (4-chloro-6-bromophenol) or other phenolic antibacterial compounds; alkyl hydroxybenzoates; cationic Antimicrobial peptides; aminoglycosides; quinolones; lincosamides; penicillins: cephalosporins; macrolides; tetracyclines; other antibiotics known in the art; , Silver colloidal antimicrobial agent, tin or copper based antimicrobial agent, manuka oil, oregano, thyme, rosemary, or other herbal extract, and grapefruit seed extract. , And the surfactants, diluents or carriers, buffers and / or bleaches described above and herein.

  Particulate BT compounds prepared using hydrophobic thiols (eg thiochlorophenol) are used, which have a greater capacity than low hydrophobic BT compounds to adhere to concrete surfaces, especially those exposed to water. Can show. BT compounds having a net negative charge, such as those having a 1: 2 molar ratio (bismuth to thiol) may also have favorable adhesive properties.

  Particulates BT in rubber, silicone and plastic products Certain embodiments include particulates described herein in or on engineered surfaces comprising processed natural and synthetic rubbers and / or rubber coatings, such as silicones and silicone coatings. BT compounds are mixed, for example, medical devices (catheters, stents, Foley catheters and other urinary catheters, gastrostomy tubes, feeding tubes, etc.), orthopedic devices, dental devices, industrial devices, electronic devices, surfaces, For example, automobiles, tires, doors and window frames, water pipes, belts, mats, attenuators (vibration mounts), trains, airplanes, ships, boats, submarines, piles, pipes, pipelines, pipes and textiles, depth measurement / water Fixtures, household goods, flooring, footwear products, exercise equipment, mobile phones, computer equipment, etc. Compounds that use organic fillers, outdoor products such as roofing materials, sunshades, tarpaulins, roof waterproofing and swimming pool linings, and, for example, sterile products, and food and beverage preservatives, pharmaceuticals, and chemicals and It is intended to reduce such rubber-surface biofilms and bioadhesives in systems for water sterilization.

  The particulate BT compounds described in the present invention can be used to integrate these and other natural and artificial materials by integrating the BT compositions and methods described herein with processing steps known for these product classes. It can be mixed in rubber products. As a non-limiting example by way of illustration, BT (but not the particulate BT described in this invention) has been incorporated into hydrogel-coated polyurethane rods and Dacron grafts (Domenico et al. Antimicrob Agents Chemother 2001; 45: 1417- 1421; Domenico et al., Peptides 2004; 25: 2047-53). WO / 2002/077095 and Japanese Patent Application No. 1997-342076 describe pre-cured and / or vulcanized raw rubber formulations containing a silver base compound to provide antimicrobial properties; 6,448,306, 6,555,599, 6,638,993, 6,848,871, 6,852,782, 6,943,205 and 7, 060,739 teaches the use of silver based antimicrobial agents in rubber matrices. The drug eluting silicone composition may comprise an antimicrobial agent dispersed in a modified biologically active binder that can be applied to a medical device or other surface without the use of an inert polymeric carrier ( US Patent Application No. 2009/0043388).

  Silicone oils generally have a molecular weight in the range of 2,000 to 30,000 and a viscosity in the range of 20 to 1,000 centistokes. Silicone rubber generally has a molecular weight in the range of 40,000 to 100,000 and a viscosity in the range of 10 to 1,000 stalk. Silicones are typically used in a variety of materials that are subject to microbial deposits. Examples of these include sealants, caulks, greases, oils, sprays, rubbers, irrigation pipes and implants. Silicone-based anti-adhesion coatings and other antimicrobial coatings have been described, but with insufficient efficacy, insufficient durability, poor biocompatibility, loss of antimicrobial activity, short useful life, high cost Suffer from disadvantages associated with other materials and other fabrics (eg, Schultz J Fluids Eng 2004; 126: 1039-47; US Pat. No. 4,025,693; Yan & Li. Ophthalmologica 2008; 222: 245-8; U.S. Pat. No. 6,221,498; U.S. Pat. No. 7,381,751; European Patent Application EP 0506113; Sawada et al. JPRAS 1990; 43: 78-82; Tiller et al. Surface Coatings International. Part B: Coatings Transactions 2005; 88: 1-82; Junhni & Newby Proceedings Annual Meeting Adhesion Society 2005; 28: 179-181; 19: 6167; US Patent Publication No. 2009/0215924; Bayston et al. Biomaterials 2009; 30: 3167-73; Gottenbos et al. Biomaterials 2002; ek 2001; 79: 337-43). These publications describe methods for incorporating antimicrobial materials into rubber products, but of the products or methods they describe are provided by the particulate BT described herein. There is nothing to offer the benefits.

  Accordingly, embodiments of the present invention are described herein in these and similar rubber (including silicone) products and methods, and in plastics and polymerisation anti-methods, such as those referred to below. Intended to replace particulate BT. In each of these and other known processing situations, the particulate BT described herein is provided on the basis of the disclosure herein and is provided by these particulate BT instead of other antimicrobial agents. The advantages disclosed herein may provide, for example, antibacterial activity, solubility and bioavailability, anti-biofilm action, non-toxicity, enhanced antibiotic efficacy.

  BT compounds can be formulated in products at low concentrations that do not interfere with the rubber processing process, for example, to reduce biofilm and prevent deposits in or on silicone products. The particulate BT concentration (by weight) within the silicone can vary from as low as about 0.0001% to about 0.1%, depending on, for example, the intended use and properties of the silicone rubber product. The microparticles BT described herein can also be used as a coating on silicone or incorporated into a silicone gel or oil to prevent or treat biofilms on the silicone surface over time. . Silicone rubber inlet valves are described in WO / 2008/064173, which regularly exudes silicone oil, and this is the effective antimicrobial level of particulate BT as described herein. The presence in the exudate imparts anti-biofilm and / or anti-adhesion capabilities to products containing such valves or similarly shaped silicone rubber devices. The erodible oil spreads to any surface near the valve, providing a renewable source of protection over time. This shape is built up, for example, below the surface of the hull or other surface exposed to water or humidity.

  For enhanced retention of BT on the rubber surface, the particulate BT described herein can be used, for example, by using a hydrophobic thiol (eg, thiochlorophenol), which can have enhanced adhesion properties, and By including BT made to have a net negative charge (eg, a 1: 2 molar ratio of bismuth to thiol) (which may also have enhanced adhesion properties), a greater hydrophobicity based on the specific thiol moiety You can choose to retain gender. For example, the silicone material can be assembled at a temperature of 100 ° C. or less in the presence of a suitable concentration of particulate BT as described herein. Bioerodible materials allow for the gradual release of such BT at levels that interfere with biofilm formation, for example, about 1-2 ppm. In other embodiments, rubber and / or plastic components are contemplated that can gradually elute and regularly replace the particulate BT compound to prevent biofouling in various industrial systems or medical devices. .

  In certain other embodiments, and similar to those described above with respect to compositions and methods for BT incorporation into rubber (including silicone) items, the particulate BT compounds described herein are described herein. By integrating the described BT compositions and methods with processing steps known for these classes of products, they can also be incorporated into these and other plastic and polymer products.

  Non-limiting examples of use with such particulate BT-containing plastic products include medical devices, orthopedic devices, dental devices, industrial devices, electronic devices, walls, floors, ceilings, roofs, and other surfaces such as Surfaces, sprinkler heads, hair care products, depth / water fittings, homes present in and on all types of vehicles, including automobiles, trains, airplanes and ships, boats, submarines, piles, pipes, pipelines and textiles Miscellaneous products, flooring, footwear products, exercise equipment, mobile phones, compounds using organic fillers, outdoor products such as roofing materials, sunshades, tarpaulins, roof waterproofing layers and swimming pool linings, and food and beverage preservatives In other products, including those used in pharmaceuticals, and in pharmaceutical and chemical and water sterilization It includes the plastic coating.

  Modern plastic materials have been used since the 1930s. Plastic products are typically made from polymers and usually additives. Typical polymers include synthetic resins, styrene, polyolefins, polyamides, fluoropolymers, vinyls, acrylics, polyurethanes, cellulosics, imides, acetals, polycarbonates and polysulfones. In order to improve the physical properties of the polymer, additives such as sparing agents are often used, which serve as a nutrient source for the microorganisms. Examples of such modern plasticizers include phthalates, adipates and other esters. These and other plasticizers are particularly susceptible to bacteria and fungi, particularly in the high humidity region, resulting in microbial surface proliferation and spore growth, which may result in one or more infections, allergic reactions in humans and animals, It can cause unpleasant odors, stains, plastic embrittlement, premature product failure and other undesirable consequences.

  Modification of plastic products during or after processing steps by the introduction of anti-adherents and other anti-microbial coatings has been described, but typically insufficient efficacy, insufficient durability, insufficient Suffers from disadvantages associated with high biocompatibility, loss of antimicrobial activity, short useful life, expensive materials and other fabrics (eg, US Pat. Nos. 3,624,062; 4,086,297; 3,663,077; 3,755,224; 3,890,270; 6,495,613; 4,348,308; 5,654,330; No. 281,677; No. 6,120,790; No. 5,906,825; No. 7,419,681; No. 5,028,664; No. 6,162,487; Markarian, Plastics, Ad diives and Compounding 2009, 11: 18-22; EP927222 B1; JP08-157641; CN1528470 A; Masatoshi et al. 2006; 51: 18-23; None of the current approaches provide the advantages provided by the particulate BT described herein. Nevertheless, what is generally known to those skilled in the art is the incorporation of antimicrobial agents in or on plastic products according to the following strategy: (a) Adsorption of agents on the polymer surface (Passive or by surfactant); (b) introduction of an antimicrobial coating applied to the surface of the molding apparatus into the polymer; (c) incorporation of the polymeric matrix material into the bulk phase; (d) polymer. Covalent binding of the agent to the surface; and / or (e) mixing the antimicrobial agent with the polymer forming (eg polyurethane) component prior to the polymerization reaction to obtain a finished polymer.

  For example, the particulate BT described herein can be introduced into these and similar systems manually or automatically as gels, sprays, liquids or powders. In one embodiment, the particulate BT, for example in powder or liquid form, is involved in components for plastic processing, such as active components (eg, polymer precursors, catalysts, initiators, crosslinkers, etc.), as well as production mixtures. It is mixed with excipients (eg carrier, solvent, release agent, dye or colorant, plasticizer), which are periodically injected into the processing system. For example, a 1 mg / ml solution or suspension of particulate BT in DMSO can be periodically injected into the polymer-forming reaction solution or sprayed onto the working part of the molding unit to produce the desired anti-bio in the finished product. Film density can be achieved.

  Accordingly, these and certain embodiments disclosed herein may include one or more microparticles BT, and optionally such as synergistic or potentiating antibiotics as described herein. It is contemplated that the particulate BT composition disclosed in the present invention may further comprise such antibiotics in such products and processes.

  Non-limiting examples of bacteria in which the compositions and methods described herein may find beneficial use according to certain embodiments described herein include Staphylococcus aureus (S. aureus). MRSA (methicillin-resistant Staphylococcus aureus), Staphylococcus epidermidis, MRSE (methicillin-resistant S. epidermidis), Mycobacterium tuberculosis, Mycobacterium abium, Pseudomonas aeruginosa, drug-resistant Pseudomonas aeruginosa, Escherichia coli Enterotoxigenic Escherichia coli, enterohemorrhagic Escherichia coli, Klebsiella pneumoniae, Clostridium difficile, Helicobacter pylori, Legionella pneumophila, Enterococcus faecalis, methicillin sensitive Enterococcus faecalis, entero Kuta cloacae, Salmonella typhimurium, Proteus vulgaris, Yersinia enterocolitica, Vibrio cholere, Shigella flexneri, vancomycin-resistant enterococcus (VRE), Burkholderia cepacia group, Francisella turalensis, Bacillus Anthracis, Yersinia pestis, Pseudomonas aeruginosa, vancomycin sensitive and vancomycin resistant Enterococcus (eg, E. faecalis, E. faecium), methicillin sensitive and methicillin resistant Staphylococcus (eg, Staphylococcus aureus, S. epidermidis) and Acinetobacter baumani, Staphylococcus haemolyticus, Staphylococcus hominis, Enterococcus feci Streptococcus pyogenes, Streptococcus agalactia, Bacillus anthracis, Klebsiella pneumoniae, Proteus mirabilis, Proteus bulgaris, Yersinia enterocolitica, Stenotrohomomonas maltophilia, Streptococcus pneumoniae, Penicillin resistance Burkholderia cepacia, Burkholderia multiborance, Mycobacterium smegmatis and E. coli. Black frog is listed.

  The practice of certain embodiments of the invention, unless specified to the contrary, is conventional in the art of microbiology, molecular biology, biochemistry, cell biology, virology and immunology, which are within the skill of the art, In addition, for reference purposes, the following references are used. Such techniques are explained fully in the literature. For example, Sambrook, et al. , Molecular Cloning: A Laboratory Manual (2nd Edition, 1989): Maniatis et al. Molecular Cloning: A Laboratory Manual (1982); DNA Cloning: A Practical Approach, vol. I & II (D. Glover, ed.); Oligonucleotide Synthesis (N. Gait ed., 1984); Nucleic Acid Hybridization (B. Hames & S. Higgins ed., 1985); eds., 1984); Animal Cell Culture (R. Freshney ed., 1986); Perbal, A Practical Guide to Molecular Cloning (1984).

  Unless the context requires otherwise, throughout this specification and the claims, the language “comprise” and variations thereof, such as “comprises” and “including” It is an open and comprehensive meaning and should be interpreted as “including but not limited to”.

  Reference throughout this specification to “one embodiment” or “an embodiment” or “aspect” is intended to mean that a particular feature, structure, or characteristic described in connection with that embodiment is at least one of the present invention It is meant to be included in the embodiment. The appearances of the phrase “one embodiment” or “an embodiment” in various places throughout this specification are not necessarily all referring to the embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

  As previously mentioned, certain embodiments of the invention described herein are intended for agricultural, industrial, manufacturing and other uses of the described BT compounds (eg, BisEDT and / or BisBAL). With respect to a formulation, the formulation may in certain further embodiments comprise one or more antibiotic compounds described herein, such as amikacin, ampicillin, cefazoline, cefepime, chloramphenicol, ciprofloxy Sasin, clindamycin (or other lincosamide antibiotics), daptomycin (Cubicin®), doxycycline, gatifloxacin, gentamicin, imipenim, levofloxacin, linezolid (Zyvox®), minocycline, nafcillin, paromomycin , Rifamupin, sulf Methoxazole, tobramycin and vancomycin; or carbapenem antibiotics, cephalosporin antibiotics, fluoroquinolone antibiotics, glycopeptide antibiotics, lincosamide antibiotics, penicillinase resistant penicillin antibiotics, and / or aminopenicillin antibiotics Substances and / or aminoglycoside antibiotics such as amikacin, arbekacin, gentamicin, kanamycin, neomycin, netilmycin, paromomycin, rhodostreptomycin, streptomycin, tobramycin or apramycin, and / or lipopeptide antibiotics such as daptomycin (Cubicin®) )), Or oxazolidinone antibiotics such as linezolid (Zyvox®) It may contain a. These and related formulations are present in or on bacterial infections that may be biofilm related (in this case there are bacteria that can promote biofilm formation, but biofilms are not yet detectable) When applied to a plant or animal, such as a plant or animal or product, containing a bacterial infection or the presence of a biofilm or other bacteria and applied to a natural or artificial surface The BT compound (s) (and optionally one or more antibiotics) may be included in a suitable carrier, excipient or diluent in an effective amount as disclosed in.

  Administration or incorporation of a BT compound or salt thereof described herein in pure form or in an appropriate agricultural, manufacturing, or other composition is an acceptable administration of a drug suitable for similar use or This can be done through any of the mixing modes. Application, incorporation or administration of the composition, in a preferred embodiment, comprises direct contact of the composition with the subject plant or animal or product to be treated, which is localized in one or more locations or widely. It may be a distributed surface site, and generally refers to contacting a topical formulation with an acute or chronic site of infection (eg, a wound site on a plant surface) surrounded by, but not limited to, intact tissue. For example, certain embodiments contemplate administering the topical formulations described herein as a topical application to a damaged, scratched or damaged natural or artificial surface.

  Formulations (e.g., agricultural compositions) include the described BT compounds (e.g., RE 37,793, U.S. 6,248,371, 6,086,921, and / or 6,380). , 248, and / or compounds prepared according to the present disclosure, such as the particulate BT suspensions described herein). In one embodiment as described in Section 1, one or more desired antibiotics (eg, aminoglycoside antibiotics such as amikacin) are suitable for use in preparing a formulation, either separately or together with a BT compound. Different vehicles, dispersants, carriers, diluents or excipients, and may be prepared depending on the intended use, solid, semi-solid, gel, cream, colloid In powders, granules, ointments, solutions, wash, gels, pastes, plasters, coatings, bioadhesives, microsphere suspensions, aerosol sprays, etc. Can be formulated in the preparation.

  The compositions of these and related embodiments are formulated so that the active ingredients can be included in the composition, and in certain preferred embodiments, the BT compounds described herein are used alone or in combination. Desired antibiotics (eg carbapenem antibiotics, cephalosporins antibiotics, fluoroquinolone antibiotics, glycopeptide antibiotics, lincosamide antibiotics, penicillinase resistant penicillin antibiotics, and aminopenicillin antibiotics Or an aminoglycoside antibiotic, such as amikacin, or rifamycin, but at the desired site and optionally around the natural or artificial surface of a plant or animal (eg human) subject, or product BT compound (s) and / or antibiotics Narubutsu (s) such that the bioavailability upon administration of formulations containing, can be applied in any order simultaneously or sequentially. Certain embodiments disclosed herein include administration of BT compounds and antibiotics to such subjects or products, and / or administration, such as administration that may be simultaneous or sequential and in any order. Alternatively, although contemplated, the present invention is not so limited, and in other embodiments, clearly distinct BT compound routes of administration are contemplated as compared to antibiotic routes of administration. That is, the antibiotic may be administered by any route described herein, while the BT compound may be administered by a route that is independent of the route used for antibiotics.

  The formulations described herein provide an effective amount of disinfectant (s) (and 22 or 2 antibiotic (s)) to prevent a desired site, such as infection or biofilm formation. Deliver to the desired site.

  As mentioned above, the formulations of the present invention may take any of a wide variety of forms, such as liquids, suspensions, plasters, creams, lotions, solutions, sprays, gels, ointments, pastes, etc. And / or may be prepared to contain liposomes, micelles and / or microspheres. See, for example, US Pat. No. 7,205,003. For example, creams are well known in the art of pharmaceutical and cosmetic formulations, are mucus or semi-solid emulsions, and are either oil-in-water or water-in-oil. The cream base is washable and contains an oil phase, an emulsifier and an aqueous phase. The oil phase, also called the “inner” phase, is generally composed of petrolatum and fatty alcohols such as cetyl alcohol or stearyl alcohol. Usually, but not necessarily, the aqueous phase has a volume that exceeds the oil phase and generally contains a humectant. Emulsifiers in cream formulations are generally nonionic, anionic, cationic or amphoteric surfactants.

  A solution is a homogeneous mixture prepared by dissolving one or more chemical substances (solutes) in a liquid and dispersing the molecules of the dissolved substance in the molecules of the solvent. The solution may contain other chemicals to buffer, stabilize or store the solute. Common examples of solvents used to prepare the solution are ethanol, water, propylene glycol or any other vehicle.

  Gels are semi-solid suspension type systems. Single-phase gels are typically aqueous, but preferably contain organic macromolecules distributed substantially homogeneously throughout the carrier liquid, which also contains alcohol and any oil. Preferred “organic macromolecules”, ie gelling agents, may be chemically cross-linked polymers, such as cross-linked acrylic acid polymers, such as polymer “carbomers”, such as carboxypolyalkylenes, which are Carbopol®. (Trademark) may be obtained commercially. Also preferred in certain embodiments are hydrophilic polymers such as polyethylene oxide, polyoxyethylene-polyoxypropylene copolymers, and polyvinyl alcohol; cellulosic polymers such as hydroxypropylcellulose, hydroxyethylcellulose, hydroxypropylmethylcellulose, hydroxy It may be propyl methylcellulose phthalate and methylcellulose; gums such as tragacanth gum and xanthan gum; sodium alginate; and gelatin. To prepare a homogeneous gel, a dispersing agent such as alcohol or glycerin can be added, and the gelling agent can be dispersed by grinding, mechanical mixing or stirring, or a combination thereof.

  Ointments, which are also well known in the art, are typically semisolid preparations based on petrolatum or other petroleum derivatives. The specific ointment base used is one that provides a plurality of desired characteristics, such as emollient, as understood by those skilled in the art. As with other carriers or vehicles, an ointment base must be inert, stable, nonirritating, and nonsensitizing. Remington: The Science and Practice of Pharmacy, 19th Ed (Easton, Pa .: Mack Publishing Co., 1995), p1399-1404, ointment bases are classified into four categories: oil-based bases; Agents; emulsion bases; and water-soluble bases. Oily ointment bases include, for example, vegetable oils, fats obtained from animals, and semisolid hydrocarbons obtained from petroleum. Emulsifiable ointment bases, also known as absorbent ointment bases, contain little or no water and include, for example, hydroxystearic sulfate, anhydrous lanolin, and hydrophilic petrolatum. Emulsion ointment bases are either water-in-oil (W / O) emulsions or oil-in-water (O / W) emulsions and include, for example, cetyl alcohol, glyceryl monostearate, lanolin, and stearic acid. Preferred water-soluble ointment bases are prepared from polyethylene glycols of various molecular weights (see for example Remington's previous work).

  The paste is a semi-solid dosage form, and the active agent is suspended in a suitable base. Depending on the nature of the base, the paste is fractionated into a fat paste or a paste produced with a single phase aqueous gel. The base in the fat paste is generally petrolatum or hydrophilic petrolatum. A paste produced from a single-phase aqueous gel generally incorporates carboxymethyl cellulose or the like as a base.

  Formulations may be prepared using liposomes, micelles and microspheres. Liposomes are microscopic vesicles having one (unilamellar) or multiple (multilamellar) lipid walls comprising a lipid bilayer, and herein, one or more of the formulations described herein. Components, such as disinfectants, or certain carriers or excipients can be encapsulated and / or adsorbed onto the lipid membrane surface. The liposomal preparations herein include cationic (positive charge), anionic (negative charge), and neutral preparations. Anionic liposomes are readily available. For example, N [1- (2,3-dioleyloxy) propyl] -N, N, N-triethylammonium (DOTMA) liposomes are commercially available under the trade name Lipofectin® (GIBCO BRL, Grand Island, NY). ). Similarly, anionic and neutral liposomes are readily available, eg, from Avanti Polar Lipids (Birmingham, AL) or can be readily prepared using readily available materials. Such materials include, among others, phosphatidylcholine, cholesterol, phosphatidylethanolamine, dioleylphosphatidylcholine (DOPC), dioleylphosphatidylglycerol (DOPG), and dioleylphosphatidylethanolamine (DOPE). These materials can also be mixed with DOTMA in appropriate ratios. Methods for producing liposomes using these materials are well known in the art.

  Micelles are composed of surfactant molecules arranged so that the polar head group forms an outer spherical shell and the hydrophobic hydrocarbon chain is positioned towards the center of the sphere to form the core. Is known in the art. Since micelles are formed in an aqueous solution containing a sufficiently high concentration of surfactant, micelles are naturally obtained. Surfactants useful for forming micelles include, but are not limited to, potassium laurate, sodium octane sulfonate, sodium decane sulfonate, sodium dodecane sulfonate, sodium lauryl sulfate, sodium doxate, decyltrimethylammonium bromide , Dodecyltrimethylammonium bromide, tetradecyltrimethylammonium bromide, tetradecyltrimethylammonium chloride, dodecylammonium chloride, polyoxy-8 dodecyl ether, polyoxy-12 dodecyl ether, nonoxynol 10, and nonoxynol 30.

  Similarly, microspheres may be incorporated into the topical formulations described herein. Like liposomes and micelles, microspheres essentially encapsulate one or more components of the inventive formulation. In general, but not necessarily, they are formed from lipids, preferably charged lipids such as phospholipids. The preparation of lipidic microspheres is well known in the art.

  Various additives known to those skilled in the art may be included in the topical formulation. For example, a solvent containing a relatively small amount of alcohol may be used to solubilize certain formulation components. Examples of suitable enhancers include, but are not limited to, ethers such as diethylene glycol monoethyl ether (commercially available as Transcutol®) and diethylene glycol monomethyl ether; surfactants such as sodium laurate, sodium lauryl sulfate, cetyltrimethyl Ammonium bromide, benzalkonium chloride, Poloxamer® (231, 182, 184), Tween® (20, 40, 60, 80), and lecithin (US Pat. No. 4,783,450); Alcohols such as ethanol, propanol, octanol, benzyl alcohol, etc .; polyethylene glycol and its esters such as polyethylene glycol monolaurate (PEGML; eg US Pat. No. 4 568,343); amides and other nitrogen compounds such as urea, dimethylacetamide (DMA), dimethylformamide (DMF), 2-pyrrolidone, 1-methyl-2-pyrrolidone, ethanolamine, diethanolamine, and triethanolamine Terpenes; alkanones; and organic acids, especially citric acid and succinic acid. Azone® and sulfoxides such as DMSO and C10MSO may be used but are less preferred.

  Some skin penetration enhancers are typically their lipophilic co-enhancers, called “plasticization” enhancers, ie, molecular weights in the range of about 150-1000 daltons, about 1 weight. %, Preferably less than about 0.5 wt.%, Most preferably less than about 0.2 wt. The Hildebrand solubility parameter of the plasticization enhancer is in the range of about 2.5 to about 10, preferably in the range of about 5 to about 10. Preferred lipophilic enhancers are fatty esters, fatty alcohols, and fatty ethers. Examples of specific and most preferred fatty acid esters include methyl laurate, ethyl oleate, propylene glycol monolaurate, propylene glycerol dilaurate, glycerol monolaurate, glycerol monooleate, isopropyl n-decanoate, and octyldodecyl Miri start is mentioned. Examples of fatty alcohols include stearyl alcohol and oleyl alcohol, and examples of fatty ethers include compounds in which a diol or triol, preferably a C2-C4 alkanediol or triol, is substituted with one or two fatty ether substituents. Is mentioned. Additional penetration enhancers are known to those skilled in the art of local drug delivery and / or are described in the relevant literature. For example, Percutaneous Penetration Enhancements eds. Smith et al. (CRC Press, Boca Raton, FL, 1995).

  In addition to those previously identified, various other additives may be included in the topical formulations according to certain embodiments of the invention. These include, but are not limited to, antioxidants, astringents, fragrances, preservatives, emollients, pigments, dyes, hygroscopic agents, propellants, and sunscreens, as well as cosmetics, drugs, Or other types of materials desirable in other respects. Typical examples of optional additives included in the formulations of certain embodiments of the invention are as follows: preservatives such as sorbates; solvents such as isopropanol and propylene glycol; astringents such as Menthol and ethanol; emollients such as polyalkylene methyl glucoside; hygroscopic agents such as glycerine; emulsifiers such as glycerol stearate, PEG-100 stearate, polyglyceryl-3 hydroxylauryl ether, and polysorbate 60; sorbitol and other polyhydroxy alcohols For example, polyethylene glycol; sunscreens such as octyl methoxycinnamate (commercially available as Parsol MCX) and butylmethoxybenzoylmethane (commercially available under the trade name Parsol 1789) Antioxidants such as ascorbic acid (vitamin C), α-tocopherol (vitamin E), β-tocopherol, γ-tocopherol, δ-tocopherol, ε-tocopherol, ζ1-tocopherol, ζ2-tocopherol, η-tocopherol, and Retinol (vitamin A); essential oils, ceramides, essential fatty acids, mineral oils, wetting agents and other surfactants such as the PLURONIC® series of hydrophilic polymers available from BASF (Mt Olive, NJ), vegetable oils (eg , Soybean oil, palm oil, liquid fraction of shea butter, sunflower oil), animal oil (eg, perhydrosqualene), mineral oil, synthetic oil, silicone oil or wax (eg, cyclomethicone and dimethicone), fluorinated oil (Generally perfluoropolyether ), Fatty alcohols (eg, cetyl alcohol), and waxes (eg, beeswax, carnauba wax, and paraffin wax); skin-feel modifiers; and thickeners and structurants, such as Swellable clays and bridged carboxypolyalkylenes which may be obtained commercially under the trade name Carbopol®.

  Other additives include, by way of example, pyrrolidine carboxylic acids and amino acids; organic antimicrobial agents such as 2,4,4'-trichloro-2-hydroxydiphenyl ether (triclosan) and benzoic acid; anti-inflammatory agents such as acetylsalicylic acid and Agents such as glycyrrhetinic acid; antiseborrheic agents such as retinoic acid; vasodilators such as nicotinic acid; melanogenesis inhibitors such as kojic acid; and mixtures thereof. Other active agents that may be beneficial when included include, for example, α-hydroxy acids, α-keto acids, polymeric hydroxy acids, humectants, collagen, marine extracts, and antioxidants such as ascorbic acid (vitamin C), α-tocopherol (vitamin E) or other tocopherols such as those described above, and retinol (vitamin A), and / or suitable salts, esters, amides, or other derivatives thereof may be present. Additional agents include those that can improve oxygen supply in living tissue, as described, for example, in WO94 / 00098 and WO94 / 00109. Sunscreen agents may be included.

  Formulations of certain embodiments of the present invention may contain conventional additives such as opacifiers, fragrances, colorants, gelling agents, thickeners, stabilizers, surfactants and the like. Other agents, such as antimicrobial agents, may be added to prevent spoilage during storage, ie to inhibit the growth of microorganisms such as yeast and mold. Suitable antimicrobial agents are typically selected from methyl and propyl esters of p-hydroxybenzoic acid (eg, methyl and propylparaben), sodium benzoate, sorbic acid, imidourea, and combinations thereof.

  A topical formulation is a BT compound (eg, as substantially homogeneous microparticles as provided herein, optionally with one or more synergistic antibiotics as described herein). In addition to (in combination), it may also contain an effective amount of one or more additional active agents suitable for the particular mode of administration or incorporation.

  The pharmacologically acceptable carrier may be incorporated into the topical formulation of certain embodiments of the present invention, and may be any carrier conventionally used in the art. Examples include water, lower alcohols, higher alcohols, honey, polyhydric alcohols, monosaccharides, disaccharides, polysaccharides, sugar alcohols such as glycol (2-carbon), glycerol (3-carbon), erythritol and threitol (4-carbon), arabitol, xylitol and ribitol (5-carbon), mannitol, sorbitol, dulcitol and iditol (6-carbon), isomaltol, maltitol, lactitol and polyglycitol, hydrocarbon oil, fat, wax, Examples include fatty acids, silicone oils, nonionic surfactants, ionic surfactants, silicone surfactants, and aqueous and emulsion based mixtures of such carriers.

  Embodiments of the topical formulations of the present invention provide treatments on natural (eg, plant or animal (including human)) or artificial (eg, product) surfaces with the frequency and amount necessary to achieve the desired result. Can be applied regularly to anything that requires The frequency of treatment depends on the nature of the application, the strength of the active ingredient in a particular embodiment (eg BT compound and optionally one or more additional active ingredients, eg antibiotics such as amikacin or other antibiotics), active ingredient delivery Depends on the effectiveness of the vehicle used to do so, as well as the ease with which the formulation is removed due to environmental factors (eg, physical contact with other materials or objects, precipitation, wind, temperature).

  Typical concentrations of active substances, such as BT compounds in the compositions described herein, are for example from 0.001 to 30% by weight, based on the total weight of the composition, from about 0.01 to 5.0%. Up to, and more preferably within the range of about 0.1 to 2.0%. As a representative example, the compositions of these embodiments of the present invention may be applied to natural or artificial surfaces at a rate of about 1.0 mg / cm 2 to about 20.0 mg / cm 2. Representative examples of topical formulations include, but are not limited to, aerosols, alcohols, anhydrous bases, aqueous solutions, creams, emulsions (including either water-in-oil or oil-in-water emulsions), fats, foams, Examples include gels, hydroalcoholic solutions, liposomes, lotions, microemulsions, ointments, oils, organic solvents, polyols, polymers, powders, salts, silicone derivatives, and waxes. The formulation may include, for example, a chelating agent, conditioning agent, emollient, excipient, hygroscopic agent, protective agent, thickener, or UV absorber. Those skilled in the art will appreciate that formulations other than those listed may be used in embodiments of the present invention.

  Chelating agents can optionally be included in certain formulations and can include any natural or synthetic chemical that has the ability to bind to a divalent cation metal such as Ca 2+, Mn 2+, or Mg 2+. Examples of chelating agents include, but are not limited to, EDTA, disodium EDTA, EGTA, citric acid, and dicarboxylic acids.

  Conditioning agents may also optionally be included in certain formulations. Examples of conditioning agents include, but are not limited to, acetylcysteine, N-acetyldihydrosphingosine, acrylate / behenyl acrylate / dimethicone acrylate copolymer, adenosine, cyclic adenosine monophosphate, adenosine monophosphate, adenosine triphosphate Acid, alanine, albutene, seaweed extract, allantoin and derivatives, aloe barbadensis extract, aluminum PCA, amyloglucosidase, arbutin, arginine, azulene, bromelain, powdered buttermilk, butylene glycol, caffeine, calcium gluconate, capsaicin , Carbocysteine, carnosine, β-carotene, casein, catalase, cephalin, ceramide, chamomile recutita (matricaria) flower extract, cholecalcif Errol, cholesteryl ester, cocobetaine, coenzyme A, processed corn starch, crystallin, cycloethoxymethicone, cysteine DNA, cytochrome C, daltside, dextran sulfate, dimethicone copolyol, dimethylsilanol hyaluronate, DNA, elastin, elastin amino acid, epithelium Growth factor, ergocalciferol, ergosterol, ethylhexyl PCA, fibronectin, folic acid, gelatin, gliadin, β-glucan, glucose, glycine, glycogen, glycolipid, glycoprotein, glycosaminoglycan, glycosphingolipid, horseradish peroxidase, Hydrogenated proteins, protein hydrolysates, jojoba oil, keratin, keratin amino acids, and kinetin, lactoferri , Lanosterol, lauryl PCA, lecithin, linoleic acid, linolenic acid, lipase, lysine, lysozyme, malt extract, maltodextrin, melanin, methionine, inorganic salt, niacin, niacinamide, oat amino acid, oryzanol, palmitoyl hydrolyzed protein, pancreatin , Papain, PEG, pepsin, phospholipid, phytosterol, placental enzyme, placental lipid, pyridoxal 5-phosphate, quercetin, resorcinol acetate, riboflavin, RNA, Saccharomyces lysate extract, silk amino acid, sphingolipid, stearamide propyl betaine , Stearyl palmitate, tocopherol, tocopheryl acetate, tocopheryl linoleate, ubiquinone, Vitis vinifera (grape) seed oil, wheat Nonoic acid, xanthan gum, and zinc gluconate. As can be readily appreciated by those skilled in the art, conditioning agents other than those listed above may be combined with the disclosed compositions or the preparations provided thereby.

  In certain embodiments, the formulations described herein may optionally include one or more emollients, such as, but not limited to, acetylated lanolin, acetylated lanolin alcohol, acrylate. / C10-30 alkyl acrylate cross polymer, acrylate copolymer, alanine, seaweed extract, aloe barbadensis extract or gel, altea officinalis extract, starch aluminum octenyl succinate, aluminum stearate, apricot (Prunus armenia) Kernel oil, arginine, arginine aspartate, arnica montana extract, ascorbic acid, ascorbyl palmitate, aspartic acid, avocado (Persia gratissima) oil, barium sulfate, barrier Ingolipid, butyl alcohol, beeswax, behenyl alcohol, β-sitosterol, BHT, birch (Betula alba) bark extract, borage (borago officinalis) extract, 2-bromo-2-nitropropane-1,3-diol, nagikada (Luscus Accureas) extract, Butylene glycol, Calendula officinalis extract, Calendula officinalis oil, Calendula officinalis oil, Calendula (euphorbia serifera) wax, Canola oil, Caprylic acid / Capric acid triglyceride, Cardamom (Eretaria cardamomum) oil, Carnauba ( Copernicia serifera wax, carrageenan (Kondoras krispas), carrot (Daucus carrotta sativa) oil, castor (ricinus communis) oil, sera , Ceresin, ceteareth-5, ceteareth-12, ceteareth-20, cetearyl octoate, ceteth-20, ceteth-24, cetyl acetate, cetyl octanoate, cetyl palmitate, chamomile (antemis nobilis) oil, cholesterol, cholesterol Esters, cholesteryl hydroxystearate, citric acid, salvia (salvia sclarea) oil, cocoa (theobroma cocoa) butter, coco-caprylic / capric acid, coconut (cocos nucifera) oil, collagen, collagen amino acid, corn Mace) oil, fatty acid, decyl oleate, dextrin, diazolidinyl urea, dimethicone copolyol, dimethiconol, dioctyl adipate, dioctyl succinate, hexacaprylic acid / hexacapric acid di Ntaerythrityl, DMDM hydantoin, DNA, erythritol, ethoxydiglycol, ethyl linoleate, eucalyptus globulus oil, evening primrose (oenothera biennis) oil, fatty acid, fructose, gelatin, wild geranium oil, glucosamine, glutamic acid glucose, glutamic acid, glyceres-26 Glyceryl, glycerol, glyceryl distearate, glyceryl hydroxystearate, glyceryl laurate, glyceryl linoleate, glyceryl myristate, glyceryl oleate, glyceryl stearate, glyceryl stearate SE, glycine, stearic acid glycol, stearic acid glycol SE, Glycosaminoglycan, grape (Vitis vinifera) seed oil, hazel (Corillas Ameri Carna) nut oil, hazel (Corillas averana) nut oil, hexylene glycol, honey, hyaluronic acid, hybrid safflower (Casamas tinctorias) oil, hydrogenated castor oil, hydrogenated cocoglyceride, hydrogenated coconut oil, hydrogenated Lanolin, hydrogenated lecithin, hydrogenated palm glyceride, hydrogenated palm kernel oil, hydrogenated soybean oil, hydrogenated tallow glyceride, hydrogenated vegetable oil, hydrolyzed collagen, hydrolyzed elastin, hydrolyzed glycosaminoglycan, hydrolyzed keratin, Hydrolyzed soy protein, hydroxylated lanolin, hydroxyproline, imidazolidinyl urea, iodopropynyl butylcarbamate, isocetyl stearate, isocetyl stearoyl stearate, isodecyl oleate, isopropyl isostearate, lano Isopropyl acid, isopropyl myristate, isopropyl palmitate, isopropyl stearate, isostearamide DEA, isostearic acid, isostearyl lactate, isostearyl neopentanoate, jasmine (jasminum officinale) oil, jojoba (baxus chinensis) oil, kelp , Kukui (Alleurites Morkana) nut oil, lactamide MEA, lanes-16, lanes-10 acetate, lanolin, lanolinic acid, lanolin alcohol, lanolin oil, lanolin wax, lavender (lavandula angstifolia) oil, lecithin, lemon ( Citrus Medica Limonam) oil, linoleic acid, linolenic acid, macadamia ternifolia nut oil, magnesium stearate, magnesium sulfate, maltitol, potassium Millet (camomile-recicita) oil, methyl glucose sesquistearate, methylsilanol PCA, microcrystalline wax, mineral oil, mink oil, maltierra oil, myristyl lactate, myristyl myristate, myristyl propionate, dicaprylic acid / neopentyl dicaprate Glycol, octyldodecanol, octyldodecyl myristate, octyldodecyl stearoyl stearate, octyl hydroxystearate, octyl palmitate, octyl salicylate, octyl stearate, oleic acid, olive (Orea Europaea) oil, orange (Citrus Aurantium)・ Durcis) oil, palm (Eraace Guinensis) oil, palmitic acid, panthetin, panthenol, panthenyl ethyl ether, paraffin, PCA, peach (Prunus persica) kernel oil, peanut (Arakis hippogae) oil, PEG-8 C12 18 ester, PEG-15 cocamine, PEG-150 distearic acid, PEG-60 glyceryl isostearate, PEG-5 glyceryl stearate PEG-30 glyceryl stearate, PEG-7 hydrogenated castor oil, PEG-40 hydrogenated castor oil, PEG-60 hydrogenated castor oil, PEG-20 methyl glucose sesquistearate, PEG-40 sorbitan peroleate, PEG- 5 soy sterol, PEG-10 soy sterol, PEG-2 stearate, PEG-8 stearate, PEG-20 stearate, PEG-32 stearate, PEG-40 stearate, PEG-50 stearate, PEG-stearate 100, PEG-150 stearate, pentadecalactone, peppermint (menta piperita) oil, petrolatum, phospholipid, polyamino sugar condensate, polyglyceryl-3 diisostearate, polyquaternium-24, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, Polysorbate 85, potassium myristate, potassium palmitate, potassium sorbate, potassium stearate, propylene glycol, propylene glycol dicaprylate / dicaprate, propylene glycol dioctanoate, propylene glycol dipelargonate, propylene glycol laurate, propylene glycol stearate, SE propylene glycol stearate, PVP, pyridoxine dipalmitate, quaternium 5, quaternium-18 hectorite, quaternium-22, retinol, retinyl palmitate, rice (Oryza sativa) nuka oil, RNA, rosemary (rosmarinus officinalis) oil, rose oil, safflower (casamas tinctorias) oil , Sage (Salvia officinalis) oil, salicylic acid, sandalwood (Santaram album) oil, serine, serum protein, sesame (Sesamu indicam) oil, shea butter (butyrospermum parky), silk powder, chondroitin sulfate sodium, DNA Sodium, sodium hyaluronate, sodium lactate, sodium palmitate, sodium PCA, sodium polyglutamate, sodium stearate, water-soluble collagen, sorbic acid, sorbitan laurate, o Sorbitan inmate, sorbitan palmitate, sorbitan sesquioleate, sorbitan stearate, sorbitol, soybean (glycine soya) oil, sphingolipid, squalane, squalene, stearamide stearamide MEA, stearic acid, stearoxydimethicone, stearoxytrimethylsilane, Stearyl alcohol, stearyl glycyrrhetinate, stearyl heptanoate, stearyl stearate, sunflower (Helianthus anus) seed oil, sweet almond (Prunus amygdalus durcis) oil, synthetic beeswax, tocopherol, tocopheryl acetate, tocopheryl linoleate, tribehenine, Tridecyl neopentanoate, tridecyl stearate, triethanolamine, tristearin, urea, vegetable oil, water, Wax, wheat (Triticum vulgare) germ oil, and ylang ylang (kananga odorata) oil.

  Surfactants may also be included, if desired, in the specific formulations contemplated herein, cationic, anionic, zwitterionic or nonionic surfactants, or their Any natural or synthetic surfactant suitable for use in cosmetic compositions, such as a mixture, can be selected (Rosen, M., “Surfactants and Interfacial Phenomena,” Second Edition, John Wiley & Sons, New Y.). , 1988, Chapter 1p431). Examples of cationic surfactants include, but are not limited to, DMDAO or other amine oxides, long chain primary amines, diamines, polyamines and their salts, quaternary ammonium salts, polyoxyethylenated Mention may be made of long chain amines, as well as quaternized polyoxyethylenated long chain amines. Examples of anionic surfactants include, but are not limited to, SDS; carboxylic acid salts (eg, soaps); sulfonic acid salts, sulfuric acid salts, phosphoric acid and polyphosphate esters; alkyl phosphates; Mention may be made of salts of perfluorocarboxylic acids. Examples of zwitterionic surfactants are cocamidopropyl hydroxysultain (CAPHS) and others that are pH sensitive and require special care in designing the optimum pH of the formulation (ie , Alkylaminopropionic acid, imidazoline carboxylate, and betaine) or those that are not pH sensitive (for example, sulfobetaine, sultaine). Examples of nonionic surfactants include, but are not limited to, alkylphenol ethoxylates, alcohol ethoxylates, polyoxyethylenated polyoxypropylene glycols, polyoxyethylenated mercaptans, long chain carboxylic acid esters, alkanols Mention may be made of amides, tertiary acetylene glycols, polyoxyethylenated silicones, N-alkylpyrrolidones, and alkyl polyglycosidases. Wetting agents, mineral oils or other surfactants such as non-ionic detergents or agents such as one or more of the PLURONIC® series (BASF, Mt Olive, NJ) can also be used, for example, according to non-limiting theory: It may be contained in order to prevent aggregation of BT fine particles in the fine particle suspension. Any combination of surfactants is acceptable. Certain embodiments are at least one anionic surfactant and at least one cationic surfactant that are compatible, ie, do not form a appreciable precipitate upon mixing, or It may comprise at least one cationic surfactant and at least one zwitterionic surfactant.

  Examples of thickeners that may also be present in certain topical formulations include, but are not limited to, acrylamide copolymer, agarose, amylopectin, bentonite, calcium alginate, carboxymethylcellulose calcium, carbomer, carboxymethylchitin, cellulose gum, dextrin , Gelatin, hydrogenated tallow, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropyl starch, magnesium alginate, methylcellulose, microcrystalline cellulose, pectin, various PEGs, polyacrylic acid, polymethacrylic acid, polyvinyl alcohol, various PPGs, sodium acrylate copolymer, carrageenan sodium, xanthan gum, and yeast β-glucan And the like. Thickeners other than those listed above may be used in embodiments of the present invention.

  According to certain embodiments contemplated herein, the BT formulation may include one or more sunscreens or UV absorbers. Where ultraviolet (UVA and UVB) absorption properties are desired, such agents include, for example, benzophenone, benzophenone-1, benzophenone-2, benzophenone-3, benzophenone-4, benzophenone-5, benzophenone-6, benzophenone-7, Benzophenone-8, benzophenone-9, benzophenone-10, benzophenone-11, benzophenone-12, benzyl salicylate, butyl PABA, cinnamate, cinoxalate, DEA-methoxycinnamate, diisopropyl methylcinnamate, ethyl dihydroxypropyl PABA, diisopropylcinnamic acid Ethyl, ethyl methoxycinnamate, ethyl PABA, ethyl urocanate, dimethoxycinnamate glyceryl octanoate, glyceryl PABA, salicylic acid glyco Le, homosalate, p- methoxycinnamic acid isoamyl, titanium, zinc, zirconium, silicon, oxides of manganese and cerium, PABA, PABA esters, Parsol 1789, and isopropylbenzyl salicylate benzyl, and may include mixtures thereof. Those skilled in the art will appreciate that sunscreens and UV absorbers or protectants other than those listed may be used in certain embodiments of the invention.

  The BT formulations disclosed herein are typically effective at pH values between about 2.5 and about 10.0. Preferably, the pH of the composition is in the following pH range or approximate values thereof: about pH 5.5 to about pH 8.5, about pH 5 to about pH 10, about pH 5 to about pH 9, about pH 5 to about pH 8, about pH 3 To about pH 10, about pH 3 to about pH 9, about pH 3 to about pH 8, and about pH 3 to about pH 8.5. Most preferably, the pH is from about pH 7 to about pH 8. One skilled in the art may add appropriate pH adjusting ingredients to the compositions of the present invention to adjust the pH to an acceptable range. If it is recognized that the pH can vary from the original value depending on the formulation and storage conditions of the formulation, the specific pH with “about” is the specific measured pH at any given time. One of ordinary skill in the art will understand that it includes formulations that can go up or down by no more than 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, or 0.1 pH units. I will.

  Creams, lotions, gels, ointments, pastes, etc. may be spread over the affected surface and gently rubbed. The solution may be applied in the same manner, but more typically it will be applied using a dropper, swab, etc. and will be carefully applied to the affected area. This application regimen will depend on many factors that can be readily determined, such as the severity of the infection and responsiveness to initial treatment. One skilled in the art can readily determine the appropriate amount of formulation to be administered, the method of administration and the rate of repetition. In general, the formulations of these and related embodiments of the invention will be applied in a range from 1 or 2 or more times per week to 1, 2, 3, 4 or more times per day. Is contemplated.

  As also previously discussed, the BT formulations useful herein may also contain an acceptable carrier, including any suitable diluent or excipient, and as such can contain the composition. It can be administered without undue toxicity, including any agent that is not harmful to the subject being administered (eg, a plant or animal, including humans). Acceptable carriers include, but are not limited to, liquids such as water, saline, glycerol and ethanol, and viscosity enhancers (eg, balsam fir resin) or film forming agents such as collodion or nitrocellulose solutions. May be included. A thorough discussion of pharmaceutically acceptable carriers, diluents, and other excipients is given in REMINGTON'S PHARMACEUTICAL SCIENCES (Mack Pub. Co., NJ, latest edition).

  The BT formulation can include an agent that binds to the BT compound, thereby assisting in delivering or holding it to the desired site on the subject or product. Suitable agents that can act in this capacity include inclusion agents such as cyclodextrins; other agents can include proteins or liposomes.

  The BT formulation is administered, applied and mixed in an effective amount, which depends on various factors such as the nature of the delivery site (if relevant), the activity of the particular BT compound used (aminoglycoside antibiotics, Including or not including an antibiotic formulation such as amikacin); metabolic stability and duration of action of the compound; (animal, including plants or humans) condition of the subject or product; mode of administration and time; excretion Varies; depending on the rate; the rate of loss of the BT compound in the normal course of activity experienced by the subject or product; and other factors. In general, the therapeutically effective daily dose is about 0.001 mg / kg (ie 0.07 mg) to about 100 mg / kg (ie 7.0 g) (for a 70 kg mammal); preferably the therapeutically effective dose is From about 0.01 mg / kg (ie, 7 mg) to about 50 mg / kg (ie, 3.5 g); more preferably a therapeutically effective dose (in a 70 kg mammal) From about 1 mg / kg (ie 70 mg) to about 25 mg / kg (ie 1.75 g). Effective plant doses can be expected to be about 10, 20, 50, or 75% lower or more.

  Effective dose ranges provided herein are not limiting and represent preferred dose ranges. However, the most preferred dosage will be adapted to each subject as understood and can be determined by one of ordinary skill in the relevant art (eg, Berkow et al., Eds., The Merck Manual 16th edition, Mark and Co., Rahway, N. J., 1992; Goodman et al., Eds. ; Avery's Drug Treatment: Principles and Practice of Clinical pharmacology and Therapeutics, 3rd edition, ADIS Press, Ltd., Williams and Wilkins, Baltimore, MD. (1987), Ebadi, Pharmacology, r. ′ S Pharmaceutical Sciences, 18th edition, Mack Publishing Co., Easton, PA (1990); Katzung, Basic and Clinical Pharmacology, Appleland and CT and NW99. ) Reference).

If desired, the total dose required for each treatment can be administered within the day by repeated or single administration. Certain preferred embodiments contemplate a single application of the BT formulation per day, per week, per 10 days, per 14 days or longer. In general, in different embodiments, treatment may be initiated with smaller dosages which are less than the optimum dose of the compound. Thereafter, the dosage is increased by small increments until the optimum effect under the circumstances is reached.
Bismuth-thiol for protection of plants and agricultural products

  Certain embodiments disclosed herein relate to compositions and methods for protecting plants and flowers from microbial infection and invasion, eg, biofilm, to reduce withering and increase product life.

  According to certain embodiments described herein, such as those summarized above, a method for protecting a plant against bacterial, fungal or viral pathogens comprising: (i) a bacterial, fungal or Prevention of plant infection by viral pathogens, (ii) inhibition of cell viability or cell proliferation of virtually all planktonic cells of bacterial, fungal or viral pathogens, (iii) biofilm formation by bacterial, fungal or viral pathogens In conditions and time sufficient for one or more of (iv) and (iv) inhibiting biofilm viability or biofilm growth of substantially all biofilm-type cells of a bacterial, fungal or viral pathogen A method comprising contacting a plant with an effective amount of a BT composition, wherein the BT composition comprises a microparticle containing a BT compound. To include a monodisperse suspension, the fine particles, a method is provided having a volume average diameter of from about 0.5μm~ about 10 [mu] m.

  In certain embodiments, the bacterial pathogen comprises Erwinia amylobora cells, and in certain embodiments, the bacterial pathogen is Erwinia amylobora, Xanthomonas campestris pathogenic type dieffenbachiae, Pseudomonas syringae, Xylella pastiniosa; Xylophilus ampelinus; • selected from Fructicola, Pantoea Stewartii subsp. Stewartii, Ralstonia solanacealum and Krabibacter mysiganensis subsp. In certain embodiments, the bacterial pathogen exhibits antibiotic resistance, and in certain other embodiments, the bacterial pathogen exhibits streptomycin resistance. In some embodiments, the plant is an edible crop plant, which in some further embodiments is a fruit tree, which in further embodiments is selected from apple, pear, peach, nectarine, plum, apricot tree. In some embodiments, the food crop plant is a Bamboo banana tree. In certain other embodiments, the food crop plant is a plant selected from tuberous plants, legumes, and grain plants. In a further embodiment, the tuberous plant is selected from potatoes and sweet potatoes.

  In certain embodiments, the contacting step is performed one or more times. In certain embodiments, at least one of the contacting steps includes one of spraying the plant, dipping the plant, coating the plant, and painting the plant. In certain embodiments, at least one step of contacting is performed at a plant flower bud, shoot or growth site, or other plant part, such as a root, bulb, stem, leaf, branch, vine, toothpick, It is carried out on or in buds, flowers or parts thereof, shoots, fruits, seeds, buds and the like. In certain embodiments, at least one step of contacting is performed within 24, 48 or 72 hours of the first flowering in the plant. In certain embodiments, the BT composition comprises BisBAL, BisEDT, Bis-dimercaprol, Bis-DTT, Bis-2-mercaptoethanol, Bis-DTE, Bis-Pyr, Bis-Ery, Bis-Tol, Bis-BDT. Bis-PDT, Bis-Pyr / Bal, Bis-Pyr / BDT, Bis-Pyr / EDT, Bis-Pyr / PDT, Bis-Pyr / Tol, Bis-Pyr / Ery, Bismuth-1-mercapto-2-propanol And one or more BT compounds selected from Bis-EDT / 2-hydroxy-1-propanethiol.

  In certain embodiments of the above methods, the methods involve synergistic or potentiating antibiotics in association with the step of contacting the plant with the BT composition, simultaneously or sequentially, and in any order. Further comprising contacting with. In some further embodiments, the synergistic or potentiating antibiotic is an aminoglycoside antibiotic, a carbapenem antibiotic, a cephalosporin antibiotic, a fluoroquinolone antibiotic, a penicillinase resistant penicillin antibiotic, and an aminopenicillin antibiotic. Contains antibiotics selected from substances. In certain embodiments, the synergistic or potentiating antibiotic is an aminoglycoside antibiotic selected from amikacin, arbekacin, gentamicin, kanamycin, neomycin, netilmicin, paromomycin, rhodostreptomycin, streptomycin, tobramycin and apramycin.

In another embodiment, a method for overcoming antibiotic resistance in a plant in which there is an antibiotic resistant bacterial phytopathogen, comprising: (a) one or more of the following: Contacting the plant with an effective amount of the BT composition for sufficient conditions and time for: (i) prevention of infection of the plant by an antibiotic-resistant bacterial pathogen, (ii) substantially all of the antibiotic-resistant bacterial pathogen Inhibition of cell viability or cell proliferation of planktonic cells, (iii) inhibition of biofilm formation by antibiotic resistant bacterial pathogens, and (iv) bio-type cell bioactivity of virtually all antibiotic resistant bacterial pathogens Inhibition of film viability or biofilm growth (wherein the BT composition comprises a substantially monodispersed suspension of microparticles comprising a BT compound, Has a volume average diameter of about 0.5 μm to about 10 μm), and (b) simultaneously or sequentially, and in any order, in connection with contacting the plant with the BT composition Contacting a plant with a synergistic or potentiating antibiotic is provided.
Disinfectant based on bismuth-thiol (BT)

  As noted above, numerous natural products (eg, antibiotics) and synthetic chemicals that have antimicrobial (antibacterial, antiviral, antifungal) properties, particularly antibacterial properties, are known in the art, and at least one Part by chemical structure, as well as antimicrobial action, eg ability to kill microorganisms (“killing” action, eg bactericidal properties), ability to stop and deplete microorganism growth (“static” action, eg bacteriostatic properties) ), Or the ability to interfere with microbial function such as colonization or infecting certain sites, bacterial secretion of exopolysaccharides, and / or conversion from planktonic populations to biofilm populations, or the expansion of biofilm formation Is characterized. Antibiotics, bactericides, disinfectants, etc. (including bismuth-thiol or BT compounds) are factors that influence the selection and use of such compositions, such as bactericidal or bacteriostatic potential, effective concentrations, and host tissues. This is discussed above and in US Pat. No. 6,582,719, including the risk of toxicity to

  Bismuth-thiol (BT), as well as related thiol compounds in which bismuth is substituted with different Group V metals (eg, arsenic, antimony) are discussed above. Compositions and methods relating to beneficial particulate BT composition particulates having a volume average diameter of about 0.5 μm to about 10 μm are also discussed herein. Accordingly, certain exemplary embodiments relate to the use of the antimicrobial agents described herein, eg, anti-biofilm agents, to treat or prevent infection and biofilm in plants. The agent is typically present in a composition containing one or more particulate bismuth-thiols in an alkaline form at a concentration of 0.0001% to 0.001% by weight. The composition may comprise BT and one or more carriers or excipients and / or further comprise other ingredients, such as other compatible fungicides, and in certain preferred embodiments described herein. Synergistic or potentiating antibiotics.

  Target crops to be protected within one intended but non-limiting embodiment include, for example, the following plant species: cereals (eg, wheat, barley, rye, oats, rice, sorghum and related) Crops), beets (eg sugar beet and feed beet), pears, drupes and soft fruits (eg apples, pears, plums, peaches, almonds, cherries, strawberries, raspberries and blackberries), legumes (eg Kidney bean, lentil, pea, soybean), oil plant (eg, rape, mustard, poppy, olive, sunflower, coconut palm, castor bean, peanut), cucurbitaceae (eg, cucumber, pumpkin, melon), Fiber plants (eg cotton, flax, Asa, jute), citrus fruits (eg Range, lemon, grapefruit, mandarin orange), vegetables (eg spinach, lettuce, asparagus, cabbage, carrot, onion, tomato, potato, paprika), camphoraceae (eg avocado, cinnamon, camphor), and other plants Corn, tobacco, nuts, coffee, sugar cane, tea, vines, hops, bananas and natural rubber plants, and ornamental plants (Asteraceae) such as flowering plants and their cut flowers. Thus, certain embodiments provide that a crop item is contacted with a composition comprising one or more of the particulate BT compounds as provided herein, such as a target crop item to be harvested, such as a cut flower or Extend the product life of target crop-derived foods (eg fruits, vegetables, grains, seeds, etc.) (eg, compared to a control group where the item is not contacted with the particulate BT described in the present invention) In addition, it is intended to statistically significantly extend the period of nutritionally and / or aesthetically useful.

  The effective concentration of particulate BT as described herein for use in these and related embodiments is derived from a number of factors, such as BT, pH, temperature, molar ratio of BT components, and harmful microorganisms. It depends on what is selected. Effectiveness also depends on whether prevention of infection or treatment of an existing infection (biofilm) is the goal of a particular application. A preventive dose is sufficient in most cases. The effective sustained concentration of BT appears to be approximately the MIC of most resistant organisms. This concentration appears to be in the range of about 1-2 μg / ml, but may be 8 μg / ml or more depending on the particular particulate BT compound (s). In one exemplary embodiment, particulate Bis pyrithione (BisPyr) is provided in a 5: 1 molar ratio (bismuth to pyrithione) for application to plants. In another embodiment, binary bismuth-thiol BisPys / Ery (bis-pyrithione / dithioerythritol) in particulate form can be provided as a broad spectrum antimicrobial agent. In yet another embodiment, the particulate BT is combined with a specific antibiotic as provided herein, preferably a synergistic or potentiating antibiotic to target against microbial infections on plants and cut flowers / trees. Can provide a unified and powerful defense. Based on the observed synergy between BisEDT and gentamicin, this BT-antibiotic combination is selected in certain embodiments for agricultural applications.

  In other embodiments, the addition of sodium bicarbonate (sodium bicarbonate) or other alkaline substance (s) (eg, potassium bicarbonate, calcium carbonate) to the particulate BT formulation adds to the antimicrobial action of BT. Or you can strengthen it. Other ingredients in the particulate BT formulation for agricultural use include surfactants and other antimicrobial agents such as chlorhexidine, sanguinarine extract, metronidazole, quaternary ammonium compounds (eg cetylpyridinium chloride) Bisguanides (eg chlorhexidine digluconate, hexetidine, octenidine, alexidine); and halogenated bisphenol compounds (eg 2,2'-methylenebis- (4-chloro-6-bromophenol) or other phenolic antibacterial compounds Alkyl hydroxybenzoates; cationic antimicrobial peptides; aminoglycosides; quinolones; lincosamides; penicillins: cephalosporins; macrolides; tetracyclines and other antibiotics; Lulutam, A-dec ICX, Coleus forskori essential oil; silver or silver colloid antimicrobial agent; tin or copper based antimicrobial agent; chlorine or bromine oxidizing agent, manuka oil; oregano; thyme; rosemary; Other herbal extracts; and grapefruit seed extract; anti-inflammatory or antioxidants such as ibuprofen, flurbiprofen, aspirin, indomethacin, aloe vera, turmeric, olive leaf extract, clove, panthenol, retinol, omega- 3 fatty acids, gamma linolenic acid (GLA), green tea, ginger, grape seed, etc .; pharmaceutically acceptable carriers such as starch, sucrose, water or water / alcohol system, DMSO, etc .; surfactants such as anionic, non-ionic Ionic, cationic and zwitterionic On- or amphoteric surfactants, or saponins derived from plant material (see, eg, US Pat. No. 6,485,711); buffers and salts; and other ingredients that may be included, such as bleaching agents, eg, peroxy compounds Potassium peroxydiphosphate; foaming systems such as sodium bicarbonate / citric acid system.

  Particulate BT compositions for agricultural and plant uses may be added, enhanced or synergistic in certain embodiments, as described herein, or in liposomal or nanoparticulate form. These and optionally other agents that produce an effect can also be combined to enhance activity and delivery. Certain embodiments explicitly exclude particulate BT formulations including liposomes, such as phospholipids (eg, phosphocholine) and / or cholesterol-containing liposomes, while certain other embodiments are not so limited, These and other liposomes can be included. Microparticles BT containing carriers, excipients or other additives that facilitate adhesion of the formulation to the surface (eg glucose, starch, citric acid, carrier oil, emulsion, dispersion, surfactant, etc.) Specific formulations can also be produced.

  In other contemplated embodiments, particulate BT formulations for use as anti-biofilm agents on plants or crops can be combined with other agents to control biofilm development. For example, it is known that interspecies quorum sensing is associated with biofilm formation. Certain agents that increase LuxS-dependent pathways or interspecies quorum sensing signals, such as N- (3-oxododecanoyl) -L-homoserine lactone (OdDHL) blocking compounds and N-butyryl-L-homoserine lactone ( BHL) analogs (eg, US Pat. Nos. 7,427,408 and 6,445,031) are useful for controlling biofilms. These anti-biofilm agents in combination with the particulate BT described herein can be delivered by leaf spray for the inhibition of bacterial biofilm development and for the treatment of pre-formed biofilms. In another embodiment, these anti-biofilm agents are contained in biodegradable microparticles for controlled release and / or in liposomal form with other antimicrobial agents.

  Thus, the particulate BT described in the present invention can be used with other current technologies to improve anti-biofilm action, according to certain embodiments. The microparticles BT of the present invention can synergize or enhance the activity of the antibiotics streptomycin and / or gentamicin against certain plant pathogens. Streptomycin does not kill bacteria, but instead inhibits their growth, thus reducing the rate at which flower stigmas are colonized, thereby reducing subsequent growth of bacteria within the nectaries (e.g. Domenico et al. J Antimicrob Chemo 1991; 28: 801-10; Domenico et al. Research Advances in Antimicrob Agents Chemother 2003; 3: 79-85). Further benefits may arise from the use of active antibiotic spray adjuvants (eg Regulaid ™) that improve the coverage and penetration of this antibiotic sufficient to allow safe use of reduced amounts of streptomycin.

  The particulate BT of the present invention can be combined with any of the active ingredients currently used to combat agricultural and plant microbial pathogens, such as oxidizing agents, chelating agents (eg iron chelating agents), bactericides and disinfectants. Preferred combinations may be additive in terms of their anti-biofilm action, or may be potentiated or synergistic according to the present disclosure. Certain embodiments include a particulate BT composition that is formulated to be hydrophobic to enhance retention of BT on the surface, for example, by using a hydrophobic thiol (eg, thiochlorophenol) that imparts enhanced adhesion properties. Intended for things. A BT with a net negative charge (eg, a 1: 2 molar ratio of bismuth to thiol) can also have enhanced adhesion properties.

  The BT compound particulate suspension can be administered as an aqueous formulation, as a suspension or solution in an organic solvent including a halogenated hydrocarbon propellant, dispersion oil, or as a dry powder. Aqueous formulations can be aerosolized with a liquid nebulizer using hydraulic or ultrasonic atomization. Propellant-based systems can use a suitable pressurized dispenser. The dry powder can use a dry powder dispenser device, which can effectively disperse the BT-containing microparticles. The desired particle size and distribution can be obtained by selecting appropriate equipment.

  Throughout this specification, unless stated otherwise, the languages “comprise”, “comprises”, and “including” include the stated step or element, or step or element group. However, it is to be understood that it does not exclude any other steps or elements, or steps or elements. “Consisting” means including the matter following the phrase “consisting of” and is limited thereto. In other words, the phrase “consisting of” indicates that the listed elements are necessary or required, and that no other elements can exist. “Consisting essentially of” is meant to include any element listed after the phrase, other that does not interfere with or contribute to the activity or action specified in the disclosure relating to the listed element Limited to elements. In other words, the phrase “consisting essentially of” requires or lists the listed elements, but does not require other elements, depending on whether it affects the activity or action of the listed elements. , May or may not be present.

  In this specification and the appended claims, the singular forms “a”, “an”, and “the” include plural references unless indicated otherwise. The term “about” or “approximately” as used in certain embodiments herein, when present before a numerical value, plus or minus 5%, 6%, 7%, 8% or 9% Indicates the range. In other embodiments, the term “about” or “approximately”, when present before a numerical value, indicates that value plus or minus 10%, 11%, 12%, 13%, or 14% of the range. In yet another embodiment, the term “about” or “approximately”, when present before a number, ranges from that value plus or minus 15%, 16%, 17%, 18%, 19%, or 20%. Indicates.

Reference: Badireddy et al. , Biotechnol Bioengineering 2008; 99: 634-43; Badireddy et al. Biomacromolecules, 2008; 9: 3079-89; Bayston et al. , Biomaterials 2009; 30: 3167-73. Codony et al. J Applied Microbiol 2003; 95: 288-93. Domenico et al. , J Antimicrob Chemo 1991; 28: 801-810. Domenico et al. , Antimicrob Agents Chemother 1997; 41: 1697-703. Domenico et al. 1999 Infect Immun 67: 664-669. Domenico et al. , Antimicrob Agents Chemother 2001; 45: 1417-21. Domenico et al. , Research Advances in Antimicrob Agents Chemother 2003; 3: 79-85. Domenico et al. , Peptides 2004. 25: 2047-53. Domenico et al. , 2005 Antibiotics for Clinicians 9: 291-297. Dufrene, J Bacteriol 2004; 186: 3283-5. Eboigbodin et al. , Biomacromolecules 2008; 9: 686-95. Fezel LM, Baummartner LK, Peterson KL, et al. Opportunistic pathogens enriched in showerhead biofilms. PNAS 2009 (epub ahead of print). Geesey GG, Lewandowski Z, Fleming HC (eds). Biofouling and biocorrosion in industrial water systems. CRC Press, Boca Raton, FL, 1994. Huang et al. , J Antimicrob Chemother 1999; 44: 601-5; Juhni et al. Proceedings Annual Meeting Adhesion Society 2005; 28: 179-181. Omoike et al. ,
Biomacromolecules 2004; 5: 1219-30. Ouazzani K, Bentama J. et al. Bio-fouling in membrane processes: micro-organism / surface interactions, hydrodynamic detection method. Congres 2008; 220: 290-4. Ozdamar et al. , Retina 1999; 19: 122-6. Piccilillo et al. J Mater Chem 2009; 19: 6167. Reunala et al. Curr Opin Allergy Clin Immunol 2004; 4: 397-401. Romo et al. , Environ Progress 1999; 18: 107-12. Saha DC, Shahin S, Rackow EC, Astiz ME, Domenico P. 2000. Cytokine modulation by bimusmuth-ethanediol in experimental sepsis. 10th Intl. Conf. Inflamm. Res. Assoc. , Hot Springs, VA. Sawada et al. , JPRAS 1990; 43: 78-82. Schultz, J Fluids Eng 2004; 126: 1039-47. Tiller JC, Hartmann L, Schable J. Reloadable antimicrobial coatings based on amphiphilic silicon networks. Surface Coatings International Part B: Coatings Transactions 2005; 88: 1-82. Tsuneda et al. , FEMS Microbiol Lett 2003; 223: 287-92. Vu et al. Molecules 2009; 14: 2535-54. Yan et al. , Ophthalmologica 2008; 222: 245-8.
Yeo et al. , Water Sci Techno 2007; 55: 35-42.
Additional references (including plant protection and related literature): Chandler et al. , Antimicrob. Agents Chemother 1978; 14: 60-8. Choudhary et al. , Microbiol Res 2009; 164: 493-513. Cooksey, Annu Rev Phytopathol 1990; 28: 201-14. Dill K, McGown EL. The biochemistry of arsenic, bismuth and antimony, h S. et al. Patai (ed.), The chemistry of organic arsenic, antimony and bismuth compounds. John Wiley & Sons, New York, 1994, pp. 695-713. Domenico et al. , 1996 J Antimicrob Chemother 38: 1031-1040. Domenico et al. 2000 Infect Med 17: 123-127. Dow et al. Proc Natl Acad Sci USA 2003; 100: 10995-1000. Dulla et al. , PNAS 2008; 105: 3-082-7. Espinosa- Urgel et al. , Microbiol 2002; 148: 341-3. Expert, Annu Rev Phytopathol 1999; 37: 307-34. Ganguli et al. , Smart Mater. Struct. 2009; 18: 104027. Huang et al. , J Antimicrob Chemother 1999; 44: 601-5. Hung et al. J Exptl Marine Biol Ecol 2008; 361: 36-41. Johnson et al. , Annu Rev Phytopathol 1998; 36: 227-48. Kang et al. , Mol Microbiol 2002; 46: 427-37. Kavouras et al. , Inverteb Biol 2005; 122: 138-51. Koczan et al. , Phytopathol 2009; 99: 1237-44. Kumar et al. , Nature Materials 2008; 7: 236-41. Marques et al. , Phytopathol 2003; 93: S57. McManus et al. , Annu Rev Phytopathol 2002; 40: 443-65. Monier et al. Proc Natl Acad Sci USA 2003; 100: 15977-82. Norelli JL, Holleran HT, Johnson WC et al. Resistance of Geneva and other apple root-stocks to Erwinia amylovora. Plant Dis 87: 26-32. Oh et al. , FEMS Microbiology Lett 2005; 253: 185-192. Omoike et al. , Biomacromolecules 2004; 5: 1219-30. Ramey et al. Curr Opinion Microbiol 2004; 7: 602-9. Salo et al. , Infection 1995; 23: 371-7. Schultz et al. , Biofouling 2007; 23: 331-41. Siboni et al. , FEMS Microbiol Lett 2007; 274: 24-9. Sosnowski et al. , Plant Pathol 2009; 58: 621-35. Tsuneda et al. , FEMS Microbiol Lett 2003; 223: 287-92. von Bodman et al. Proc Natl Acad Sci USA 1998, 95: 7687-7692. Vu et al. Molecules 2009; 14: 2535-54. Zaini et al. , FEMS Microbiol LETT 2009; 295: 129-34.

  The following examples are given by way of illustration and are not meant to be limiting.

Example Example 1
Preparation of BT compounds The following BT compounds were prepared according to the method of Domenico et al. (US RE 37,793, US 6,248,371, 6,086,921, 6,380,248). Or as microparticles according to the synthesis protocol described below for BisEDT. Shown is the atomic ratio for a single bismuth atom as a comparison, based on the theoretical ratio of the reactants used to form the trivalent complex with the sulfur-containing compound and the known propensity of bismuth. The numbers in parentheses are the ratio of bismuth to one (or several) thiol agent (eg Bi: thiol 1 / thiol 2; see also Table 1).
1) CPD 1B-1 Bis-EDT (1: 1) BiC2H4S2
2) CPD 1B-2 Bis-EDT (1: 1.5) BiC3H6S3
3) CPD 1B-3 Bis-EDT (1: 1.5) BiC3H6S3
4) CPD 1C Bis-EDT (soluble Bi preparation) (1: 1.5) BiC3H6S3
5) CPD 2A Bis-Bal (1: 1) BiC3H6S2O
6) CPD 2B Bis-Bal (1: 1.5) BiC4.5H9O1.5S3
7) CPD 3A Bis-Pyr (1: 1.5) BiC7.5H6N1.5O1.5S1.5
8) CPD 3B Bis-Pyr (1: 3) BiC15H12N3O3S3
9) CPD 4 Bis-Ery (1: 1.5) BiC6H12O3S3
10) CPD 5 Bis-Tol (1: 1.5) BiC10.5H9S3
11) CPD 6 Bis-BDT (1: 1.5) BiC6H12S3
12) CPD 7 Bis-PDT (1: 1.5) BiC4.5H9S3
13) CPD 8-1 Bis-Pyr / BDT (1: 1/1)
14) CPD 8-2 Bis-Pyr / BDT (1: 1 / 0.5)
15) CPD 9 Bis-2 hydroxy, propanethiol (1: 3)
16) CPD 10 Bis-Pyr / Bal (1: 1 / 0.5)
17) CPD 11 Bis-Pyr / EDT (1: 1 / 0.5)
18) CPD 12 Bis-Pyr / Tol (1: 1 / 0.5)
19) CPD 13 Bis-Pyr / PDT (1: 1 / 0.5)
20) CPD 14 Bis-Pyr / Ery (1: 1 / 0.5)
21) CPD 15 Bis-EDT / 2 hydroxy, propanethiol (1: 1/1)

  Particulate bismuth-1,2-ethanedithiol (Bis-EDT, a soluble bismuth preparation) was prepared as follows:

  To an excess of room temperature 5% aqueous HNO3 (11.4 L) in a 15 L polypropylene carboy, add aqueous Bi (NO3) 3 solution (43% Bi (NO3) 3 (w / w), 5% nitric acid (w / w) ), 52% (w / w) water, Shepherd Chemical Co., Cincinnati, OH, product number 2362; δ-1.6 g / mL) 0.331 L (-0.575 mol) is slowly added dropwise with stirring. Then, absolute ethanol (4 L) was slowly added. A small amount of white precipitate formed but dissolved upon continuous stirring. Separately, 72.19 mL (0.863 mol) of 1,2-ethanedithiol (CAS 540-63-6) was added to 1.5 L of absolute ethanol using a 60 mL syringe, followed by stirring for 5 minutes. An ethanolic solution of 1,2-ethanedithiol (˜1.56 L, ˜0.55 M) was prepared. The 1,2-ethanedithiol / EtOH reagent was then slowly added dropwise to the aqueous Bi (NO3) 3 / HNO3 solution over 5 hours and continuously stirred overnight. The formed product was allowed to settle as a precipitate for approximately 15 minutes, after which the filtrate was removed at 300 mL / min using a peristaltic pump. Thereafter, the product was recovered by filtration through a filter paper in a 15 cm diameter Buchner funnel, and subsequently washed three times with 500 mL each of ethanol, USP water and acetone to give BisEDT (694.51 g / mol) as a yellow amorphous. Obtained as a fine powder solid. The product was placed in a 500 mL brown glass bottle and dehydrated with CaCl 2 under high vacuum for 48 hours. The recovered material (yield ~ 200 g) released an odor characteristic of thiols. The crude product was redissolved in 750 mL absolute ethanol, stirred for 30 minutes, then filtered, washed 3 times with 50 mL ethanol, 2 times 50 mL acetone, and washed again with 500 mL acetone. The rewashed powder was triturated in 1M NaOH (500 mL), filtered and washed 3 times with 220 ml of water, 2 times with 50 mL of ethanol and 1 time with 400 mL of acetone to give 156.74 g of purified BisEDT. . The next batch, prepared in essentially the same way, yielded a yield of about 78.91%.

  The product was obtained by analysis of the data from 1H and 13C nuclear magnetic resonance (NMR), infrared spectroscopy (IR), ultraviolet spectroscopy (UV), mass spectrometry (MS) and elemental analysis as shown above. It was characterized as having the structure of I. An HPLC method was developed to determine the chemical purity of BisEDT, thereby preparing a sample in DMSO (0.5 mg / mL). Λmax was measured by scanning a solution of BisEDT in DMSO at 190-600 nm. Waters (Millipore Corp., Milford, Mass.) Model with a UV detector monitoring at 265 nm (λmax) and equipped with an YMC Pack PVC Sil NP (inner diameter 250 × 4.6 mm) analytical column (Waters) with an injection volume of 2 μL Using a 2695 chromatograph, gradient HPLC elution at 1 mL / min at ambient temperature with 0.1% formic acid in acetone: water (9: 1) as the mobile phase, detecting a single peak, A chemical purity of 100 ± 01% was indicated. Elemental analysis was consistent with the structure of formula (I).

  The dry particulate material was characterized to evaluate particle size characteristics. Briefly, the microparticles are resuspended in 2% Pluronic® F-68 (BASF, Mt. Olive, NJ) and the suspension is sonicated for 10 minutes in a standard water bath sonicator. And then analyzed using a Nanosizer / Zetasizer Nano-S particle analyzer (model ZEN 1600 (without zeta potential measurement capability), Malverm Instruments, Worcestershire, UK) according to the manufacturer's recommendations. From the combined data of the two measurements, the microparticles show a unimodal distribution and all detectable events occur between volume mean diameter (VMD) 0.6 microns to 4 microns, peaking at about 1.3 microns Had VMD. On the other hand, when BisEDT was prepared by the prior art method (Domenico et al., 1997 Antimicrob. Agents Chemother. 41 (8): 1697-1703), the majority of the particles were heterodispersed and significantly larger in size. Characterization based on is impossible.

Example 2
Colony Biofilm Model of Chronic Wound Infection: Inhibition by BT Compounds Bacteria present in chronic wounds essentially follow the method described to adopt a biofilm lifestyle (Anderl et al., 2003 Antimicrob Agents Chemother). 47: 1251-56; Walters et al., 2003 Antimicrob Agents Chemother 47: 317; Wentland et al., 1996 Biotnol. Prog. 12: 316; Zheng et al., 2002 Antim. Using biofilm, BT can be compared with biofilm for the effect on bacterial cell survival. And inspected.

  Briefly, colony biofilms were grown on 10% trypsin soy agar for 24 hours and transferred to treatment containing Mueller Hinton plates. After the treatment, the biofilm was dispersed in peptone water containing 2% w / v glutathione (BT neutralization), serially diluted with peptone water, and then spotted on a counting plate. Two bacteria isolated from chronic wounds were used separately in generating a colony biofilm for testing. These were Pseudomonas aeruginosa Gram-negative strains and Gram-positive bacteria, methicillin-resistant Staphylococcus aureus (MRSA).

  Bacterial biofilm colonies were obtained essentially as described (Anderl et al., 2003 Antimicrob Agents Chemother 47: 1251-56; Walters et al., 2003 Antimicrob Agents Chemel 47.317 et al .; Prog. 12: 316; Zheng et al., 2002 Antimicrob Agents Chemother 46: 900), grown on microporous membranes on agar plates. Colony biofilms have shown many of the familiar properties of other biofilm models, for example, they were composed of cells that were densely aggregated in a highly hydrated matrix. As reported elsewhere (Brown et al., J Surg Res 56: 562; Millward et al., 1989 Microbios 58: 155; Such et al., 1995 J Pharm Pharmacol 47: 1094; Thrower et al. Med Microbiol 46;: 425), it was observed that the bacteria in the colony biofilm also showed significantly reduced antimicrobial susceptibility, as quantified in the higher performance in vitro biofilm reactor. The majority of colony biofilms occurred immediately and reproducibly. According to a non-limiting theory, this colony biofilm model shares some of the characteristics of infected wounds, and the bacteria are in the air with nutrients supplied from below the biofilm and a minimum amount of fluid. Developed at the interface. Various nutrient sources, such as blood agar, thought to mimic in vivo nutrient conditions, were used to culture the colony biofilm.

  Colony biofilms were prepared by inoculating a 25 mm diameter polycarbonate filter membrane with a 5 μl spot of planktonic bacterial liquid medium. Prior to inoculation, the membrane was sterilized by exposing it to UV light for 10 minutes per side. The inoculum was grown overnight in 37 ° C. bacterial medium, diluted with fresh medium to an optical density of 0.1 at 600 nm, and then attached to the membrane. The membrane was then placed on an agar plate containing a growth medium. Thereafter, the plate was covered and inverted in a 37 ° C. incubator. Every 24 hours, membranes and colony biofilms were transferred to new plates using sterile tweezers. Colony biofilms were typically used for experiments 48 hours after development, at which time there were approximately 109 bacteria per membrane. The colony biofilm method has been successfully used to culture a wide variety of single and mixed species biofilms.

  To determine susceptibility to antimicrobial agents (eg, BT compounds such as BT compound combinations; antibiotics; and BT compounds-antibiotic combinations), colony biofilms are supplemented with candidate antimicrobial treatments Transferred to agar plates. When the duration of exposure to antimicrobial treatment exceeded 24 hours, the colony biofilm was transferred daily to a new treatment plate. At the end of the treatment period, the colony biofilm was placed in a test tube containing 10 ml of buffer and vortexed for 1-2 minutes to disperse the biofilm. In some instances, the sample had to be easily treated with a tissue homogenizer to break up cell aggregates. The resulting cell suspension was then serially diluted and allowed to stand and the surviving bacteria were counted and reported as colony forming units (CFU) per unit area. Survival data was analyzed using log10 transformation.

  For each type of bacterial biofilm colony culture (Pseudomonas aeruginosa, PA; methicillin resistant Staphylococcus aureus, MRSA or SA), 5 antibiotics and 13 BT compounds were tested. Antimicrobial agents tested against PA include BisEDT and BT referred to herein as compounds 2B, 4, 5, 6, 8-2, 9, 10, 11 and 15 (see Table 1) and antibiotics And tobramycin, amikacin, imipenim, cefazolin, and ciprofloxane. Antimicrobial agents tested against SA include BisEDT and BT called compounds 2B, 4, 5, 6, 8-2, 9, 10 and 11 (see Table 1) and antibiotics rifampicin, daptomycin, minocycline, Ampicillin and vancomycin were included. Antibiotics were tested at concentrations approximately 10-400 times the minimum inhibitory concentration (MIC) according to established microbiological methodology, as described previously in the “Brief Description of the Drawings”.

  Seven BT compounds showed significant effects on PA bacterial survival at the tested concentrations, and two BT compounds demonstrated significant effects on MRSA survival at the tested concentrations; bacterial survival Representative results showing the effect of BT on BD are shown in FIG. 1 for BisEDT and BT compound 2B (tested against PA) and in FIG. 2 for BT compounds 2B and 8-2 (tested against SA) Both are relative to the effects of the indicated antibiotics. As also shown in FIGS. 1 and 2, inclusion of the indicated BT compound in combination with the indicated antibiotic provides a synergistic effect, whereby the ability to reduce bacterial survival is either antibiotic alone or BT compound It was enhanced compared to any single antimicrobial effect. Effect obtained with the combination of 1600 μg / mL AMK + 80 μg / mL BisEDT on Compound 15 (BisEDT / 2-hydroxy, propanethiol (1: 1/1)) at a concentration of 80 μg / mL in the PA survival assay (FIG. The effect comparable to 1) (not shown) was shown.

Example 3
Inhibition of chronic wound infection by drip flow biofilm model BT compounds Drip flow biofilms are recognized and reliable models for forming bacterial biofilms and testing the effects of candidate antimicrobial compounds on them. is there. The drip flow biofilm is produced on a coupon (substrate) placed in the drip flow reactor groove. Many different types of materials, such as frosted glass microscope slides, can be used as substrates for bacterial biofilm formation. The nutrient liquid medium is dropped into the cell chamber of the drip flow bioreactor near the top to place the medium into the chamber and then run at a 10 ° C. ramp over the length of the coupon.

  Biofilms are grown in a drip flow bioreactor, BT compounds alone or in combination with antibiotic compounds and / or antibiotic compounds alone or in combination with other antibacterial agents including BT compounds Or other conventional or candidate treatments for chronic wounds. Thus, the BT compound is characterized for its effect on bacterial biofilm in the drip flow reactor. Biofilms in drip flow reactors are prepared according to established methodologies (eg, Stewart et al., 2001 J Appl Microbiol. 91: 525; Xu et al., 1998 Appl. Environ. Microbiol. 64: 4035). . This design involves culturing the biofilm on a polystyrene coupon tilted in a covered chamber. An exemplary medium is 5% v / v adult donor bovine serum (pH 6.8) that mimics conditions of high serum protein and very low iron concentrations, similar to in vivo biofilm growth conditions such as chronic wounds. Contains supplemented 1 g / l glucose, 0.5 g / l NH4NO3, 0.25 g / l KCl, 0.25 g / l KH2PO4, 0.25 g / l MgSO4 · 7H2O. The medium is added dropwise (5 ml / hour) onto four coupons contained in four separate parallel chambers, each 10 cm × 1.9 cm and 1.9 cm deep. A reactor divided into chambers was assembled with polysulfone plastic. Each chamber is fitted with an individual removable plastic lid that can be tightly closed. The biofilm reactor is placed in a 37 ° C. incubator and warmed by passing the bacterial cell medium through an aluminum heating bath held in the incubator. This method mimics the antibiotic-resistant phenotype observed in certain biofilms, mimicking the low fluid shear environment and the proximity to the air interface characteristics of chronic wounds, while It is compatible with numerous analytical methods that provide continuous supplementation and characterize and monitor the effectiveness of the introduced candidate antimicrobial regimen. The drip flow reactor has been successfully used to culture a wide variety of pure and mixed species biofilms. Biofilms are typically grown for 2-5 days before an antimicrobial agent is applied.

  To measure the effect of an anti-biofilm agent on a biofilm grown in a drip flow reactor, the fluid flow passing over the biofilm is converted into a desired treatment formulation (eg, one or more BT compounds and / or Or modified or supplemented with one or more antibiotics, or controls, and / or other candidate agents). The flow is continued for a specific treatment period. The treated biofilm coupon is then briefly removed from the reactor and scraped into a beaker containing 10 ml of buffer. Samples are briefly treated with a tissue homogenizer (typically 30 seconds to 1 minute) to disperse bacterial aggregates. The suspension is serially diluted to enumerate viable microorganisms according to standard microbiological methodology.

Example 4
Inhibition of keratinocyte scratch repair by wound biofilm: Biofilm suppression by BT compounds This example modifies an established in vitro keratinocyte scratch model of wound healing to provide biofilm-related wound pathology And relevance to wound healing, in particular to reach a model with relevance to acute or chronic wounds or wound-containing biofilms as described herein. With the keratinocyte scratch model for the effects of chronic wound biofilms, the culture of mammalian (eg, human) keratinocytes and bacterial biofilm populations proceeds in separate chambers in fluid contact with each other, Allows assessment of the effect of conditions that affect the effect of soluble components assimilated by the biofilm on keratinocyte wound healing events.

  Neonatal human foreskin cells are cultured as a monolayer in a treated plastic dish in which controlled “wounds” or scratches are formed by mechanical means (eg, sterile scalpel, razor, By means of a monophasic physical disruption, such as scratching an essentially straight cell-free zone between regions of the monolayer using a suitable tool such as a cell scraper, tweezers, or other instrument). In vitro keratinocyte monolayer model systems are known to undergo cellular structural and functional processes in response to wound events in a manner that mimics wound healing in vivo. According to the embodiments disclosed herein, the effect of the presence of bacterial biofilm on such a process, for example on the healing time of the scratch, was observed and selected in these and related embodiments. The effect of the presence of candidate antimicrobial (eg, antibacterial and antibiofilm) treatments is also evaluated.

  Wounded keratinocyte monolayers cultured in the presence of biofilms are investigated by morphological, biochemical, molecular genetics, cell physiology and other parameters, and the introduction of BT compounds is Decide whether to alter the distracting effects of the film (eg, increase or decrease in a statistically significant manner relative to the appropriate control). In order to evaluate the effect of such treatment on the impact of biofilm on the model wound healing process after examining the toxicity of each BT compound treatment, the wound was first designed with each BT compound alone and BT compound. Expose to combination.

  In an exemplary embodiment, a membrane (eg, a TransWell membrane insert, etc.) retained in a previous tissue culture well and in fluid communication with a keratinocyte monolayer that is scratched to initiate the wound healing process. Above, the third day biofilm is cultured. Biofilms cultured from genuine acute or chronic wounds are contemplated for use in these and related embodiments.

  Thus, an in vitro system has been developed to evaluate the effect of soluble biofilm components on human keratinocyte migration and proliferation. The system separates biofilm and keratinocytes using a dialysis membrane. Keratinocytes are cultured from neonatal foreskin as previously described (Flekman et al., 1997 J Invest. Dermatol. 109: 36; Pieporn et al., 1987 J Invest. Dermatol. 88: 215-219) Grow as a confluent monolayer on a glass cover slip. The monolayer of keratinocytes can then be scratched to create a uniform width “wound” and then the cell repair process can be monitored (eg, Tao et al., 2007 PLoS ONE 2: e697; Buth et al., 2007 Eur. J Cell Biol. 86: 747; Phan et al., 2000 Ann. Acad. Med. Singapore 29:27). Then, using aseptic technique, the artificial wound is placed in the bottom of a sterile double-sided chamber and the chamber is assembled. A keratinocyte growth medium (EpiLife) with or without antibiotics and / or bismuth-thiol is filled on both sides of the chamber. An uninoculated system is used as a control.

  Bacteria isolated from the wound are inoculated into the system and incubated for 2 hours in static conditions to allow the bacteria to adhere to the surface in the upper chamber. Following the attachment period, liquid medium flow is initiated in the upper chamber to remove unattached cells. Thereafter, the flow of the medium is continued at a rate that minimizes the growth of the plactonic cells in the upper chamber by washing out unattached cells. After an incubation time in the range of 6 to 48 hours, the system (keratinocyte monolayer on the coverslip and bacterial biofilm on the membrane substrate) is disassembled and the coverslip is removed and analyzed. In a related embodiment, after the mature biofilm is grown in the upper chamber, the chamber is assembled. In other related embodiments, changing at least one indicator of scratch healing (e.g., statistically relative to an appropriate control), such as the time that elapses during wound repair or other signs of wound repair. Candidate agents such as BT compounds or potentially synergistic BT compounds + antibiotic combinations for keratinocyte scratch repair (to increase or decrease in a significant manner). For example, BT compounds provided herein, such as BT provided in particulate form, and amikacin, ampicillin, cefazoline, cefepime, chloramphenicol, ciprofloxacin, clindamycin (or other lincosamide antibiotics) ), Daptomycin (Cubicin®), doxycycline, gatifloxacin Biofilms and scrapes to determine the effects of gentamicin, imipenim, levofloxacin, linezolid (Zyvox®), minocycline, nafcillin, paromomycin, rifampine, sulfamethoxazole, tobramycin and vancomycin Separate co-cultures of wounded keratinocyte monolayers are performed in the absence or presence of one or more BT compounds, optionally with or without one or more antibiotics (eg, Tao et al., 2007 PLoS ONE 2: e697; Buth et al., 2007 Eur.J Cell Biol.86: 747; Phan et al., 2000 Ann. Acad. Med. Singapore 29:27).

Example 5
Inhibition of keratinocyte scratch repair by wound biofilm Isolated human keratinocytes were cultured on glass coverslips to create scratches according to the methodology described in Example 4 above. The wounded culture was maintained on a membrane support in fluid communication with the keratinocyte culture, under culture conditions alone or in the presence of a co-cultured biofilm. The time interval between scratch closure was then measured while cell growth and / or migration of keratinocytes re-established the keratinocyte monolayer throughout the scratch zone. FIG. 3 shows the effect of biofilm fluid communication (but not in direct contact) on the healing time of a scratched keratinocyte monolayer.

  Thus, in certain embodiments, scratched cell (eg, keratinocytes or fibroblasts) monolayers are cultured in the presence of a bacterial biofilm with or without a candidate anti-biofilm agent. And assessing an indication of healing of a scratched cell monolayer in the absence and presence of a candidate anti-biofilm agent, wherein the at least one indication of healing is promoted Drugs (eg, BT compounds such as the substantially monodispersed BT microparticle suspensions described herein alone or antibiotics such as amikacin, ampicillin, cefazoline, cefepime, chloramphenicol, ciprofl Loxacin, clindamycin, daptomycin (Cubicin®), doxycycline, gatifloxacin, gentamicin Acute or chronic wounds, in combination with one or more of: Or a method of identifying an agent that treats a chronic wound, identified as an appropriate agent to treat a wound comprising a biofilm.

Example 6
Synergistic Bismuth-Thiol (BT) Antibiotic Combination This example comprises one or more bismuth-thiol compounds and one or more antibiotics for various bacterial species and strains, such as multiple antibiotic-resistant bacteria. An example of the demonstrated synergistic effect of the combination is shown.

  Materials and Methods susceptibility testing, NCCLS protocol (National Committee for Clinical Laboratory Standards (1997) Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically:... Approved Standard M7-A2 and Informational Supplement M100-S10 NCCLS, Wayne, PA USA), and was performed by broth dilution in 96-well tissue culture plates (Nalge Nunc International, Denmark).

  Briefly, an overnight bacterial culture was utilized to prepare a 0.5 McFarland standard suspension, which was further 1: 1 in cation-conditioned Mueller-Hinton broth medium (BBL, Cockeysville, MD, USA). Dilute by 50 (~ 2x106 cfu / ml). BT (prepared as described above) and antibiotics were added at increasing concentrations to keep the final volume constant at 0.2 mL. Cultures were incubated at 37 ° C. for 24 hours and turbidity was assessed by absorbance at 630 nm using an ELISA plate reader (Biotek Instruments, Winooski, VT, USA) according to manufacturer's recommendations. The minimum inhibitory concentration (MIC) was expressed as the lowest drug concentration that prevented growth for 24 hours. Viable bacterial counts (cfu / mL) were determined by standard plating on nutrient agar. The minimum bactericidal concentration (MBC) was expressed as the drug concentration that reduced initial viability by 99.9% at 24 hours after inoculation.

  The activity of the antimicrobial combination was evaluated using the checkerboard method. Partial Inhibitory Concentration Index (FICI) and Partial Bactericidal Concentration Index (FBCI) were determined according to Eliopoulos et al. Of Eliopoulos and Meollering, (1996) Antimicrobial combina- tions. Synergy is defined as a FICI or FBCI index of 0.5 or less, and 0.5 to 4 or less, no interaction, and 4 or more defined as antagonistic (Odds, FC (2003) Synergy, antagonistism) , And what the chequebord puts between them.Journal of Antibiotic Chemotherapy 52: 1). As before, a reduction of more than 4 times the antibiotic concentration was also defined as synergy.

ＢＥ＝０．２μｇ／ｍｌ ＢｉｓＥＤＴ；菌株は、Ｗｉｎｔｈｒｏｐ−Ｕｎｉｖｅｒｓｉｔｙ Ｈｏｓｐｉｔａｌ， Ｍｉｎｅｏｌａ， ＮＹのＣｌｉｎｉｃａｌ Ｍｉｃｒｏｂｉｏｌｏｇｙ Ｌａｂｏｒａｔｏｒｙから得た。ナフシリンは、Ｓｉｇｍａ（Ｓｔ． Ｌｏｕｉｓ， ＭＯ）から得た。

ＢＥ＝０．２μｇ／ｍｌ ＢｉｓＥＤＴ；菌株は、Ｗｉｎｔｈｒｏｐ−Ｕｎｉｖｅｒｓｉｔｙ Ｈｏｓｐｉｔａｌ， Ｍｉｎｅｏｌａ， ＮＹのＣｌｉｎｉｃａｌ Ｍｉｃｒｏｂｉｏｌｏｇｙ Ｌａｂｏｒａｔｏｒｙから得た。ナフシリンは、Ｓｉｇｍａから得た。

ＢＥ＝０．２μｇ／ｍｌ ＢｉｓＥＤＴ；菌株Ｓ２４４６−３は、Ｗｉｎｔｈｒｏｐ−Ｕｎｉｖｅｒｓｉｔｙ Ｈｏｓｐｉｔａｌ， Ｍｉｎｅｏｌａ， ＮＹのＣｌｉｎｉｃａｌ Ｍｉｃｒｏｂｉｏｌｏｇｙ Ｌａｂｏｒａｔｏｒｙから得た。抗体は、Ｓｉｇｍａから得た。

ＧＭ＝ゲンタマイシン；菌株Ｓ２４００−１は、Ｗｉｎｔｈｒｏｐ−Ｕｎｉｖｅｒｓｉｔｙ Ｈｏｓｐｉｔａｌ， Ｍｉｎｅｏｌａ， ＮＹのＣｌｉｎｉｃａｌ Ｍｉｃｒｏｂｉｏｌｏｇｙ Ｌａｂｏｒａｔｏｒｙから得た。ゲンタマイシンは、ＷｉｎｔｈｒｏｐのＰｈａｒｍａｃｙ Ｄｅｐａｒｔｍｅｎｔから得た。太字は相乗性。

μｇ／ｍｌでのデータ；菌株Ｓ２４００−１は、Ｗｉｎｔｈｒｏｐ−Ｕｎｉｖｅｒｓｉｔｙ Ｈｏｓｐｉｔａｌ， Ｍｉｎｅｏｌａ， ＮＹのＣｌｉｎｉｃａｌ Ｍｉｃｒｏｂｉｏｌｏｇｙ Ｌａｂｏｒａｔｏｒｙから得た。抗生物質は、ＷｉｎｔｈｒｏｐのＰｈａｒｍａｃｙ Ｄｅｐａｒｔｍｅｎｔから得た。

μｇ／ｍｌでのデータ；菌株Ｓ２４００−１は、Ｗｉｎｔｈｒｏｐ−Ｕｎｉｖｅｒｓｉｔｙ Ｈｏｓｐｉｔａｌ， Ｍｉｎｅｏｌａ， ＮＹのＣｌｉｎｉｃａｌ Ｍｉｃｒｏｂｉｏｌｏｇｙ Ｌａｂｏｒａｔｏｒｙから得た。抗生物質は、ＷｉｎｔｈｒｏｐのＰｈａｒｍａｃｙ Ｄｅｐａｒｔｍｅｎｔから得た。

μｇ／ｍｌでのデータ；菌株Ｓ２４００−１は、Ｗｉｎｔｈｒｏｐ−Ｕｎｉｖｅｒｓｉｔｙ Ｈｏｓｐｉｔａｌ， Ｍｉｎｅｏｌａ， ＮＹのＣｌｉｎｉｃａｌ Ｍｉｃｒｏｂｉｏｌｏｇｙ Ｌａｂｏｒａｔｏｒｙから得た。抗生物質は、ＷｉｎｔｈｒｏｐのＰｈａｒｍａｃｙ Ｄｅｐａｒｔｍｅｎｔから得た。

μｇ／ｍｌでのデータ；抗生物質は、Ｗｉｎｔｈｒｏｐ−Ｕｎｉｖｅｒｓｉｔｙ Ｈｏｓｐｉｔａｌ， Ｍｉｎｅｏｌａ， ＮＹのＰｈａｒｍａｃｙ Ｄｅｐａｒｔｍｅｎｔから得た。

ＡＢ＝抗生物質；ＣＭ＝クロラムフェニコール；ＡＭ＝アンピシリン；ＢＥ＝０．３μｇ／ｍｌのＢｉｓＥＤＴ、菌株は、Ｄｅｐａｒｔｍｅｎｔ ｏｆ Ｍｏｌｅｃｕｌａｒ Ｇｅｎｅｔｉｃｓ ａｎｄ Ｃｅｌｌ Ｂｉｏｌｏｇｙ，Ｔｈｅ Ｕｎｉｖｅｒｓｉｉｔｙ ｏｆ Ｃｈｉｃａｇｏ， Ｃｈｉｃａｇｏ，ＩＬのＤｒ． ＭＪ Ｃａｓａｄａｂａｎの研究室から得た。抗生物質は、Ｗｉｎｔｈｒｏｐ−Ｕｎｉｖｅｒｓｉｔｙ Ｈｏｓｐｉｔａｌ， Ｍｉｎｅｏｌａ， ＮＹのＰｈａｒｍａｃｙ Ｄｅｐａｒｔｍｅｎｔから得た。

ＤＯＸ＝ドキシサイクリン；ＢＥ＝０．３μｇ／ｍｌのＢｉｓＥＤＴ；菌株は、Ｄｅｐａｒｔｍｅｎｔ ｏｆ Ｂａｃｔｅｒｉｏｌｏｇｙ， Ｔｈｅ Ｕｎｉｖｅｒｓｉｔｙ ｏｆ Ｂｒｉｓｔｏｌ， Ｂｒｉｓｔｏｌ， ＵＫのＤｒ．Ｉ Ｃｈｏｐｒａの研究室から得た。抗生物質は、Ｗｉｎｔｈｒｏｐ−Ｕｎｉｖｅｒｓｉｔｙ Ｈｏｓｐｉｔａｌ， Ｍｉｎｅｏｌａ， ＮＹのＰｈａｒｍａｃｙ Ｄｅｐａｒｔｍｅｎｔから得た。

Ａｇｒ＝アミノグリコシド耐性；ＮＮ＝トブラマイシン；ＰＡ＝シュードモナス・エルギノーサ；ＢＥ＝ＢｉｓＥＤＴ、０．３μｇ／ｍｌ；菌株は、Ｄｅｐａｒｔｍｅｎｔ ｏｆ Ｍｉｃｒｏｂｉｏｌｏｇｙ ａｎｄ Ｉｍｍｕｎｏｌｏｇｙ， Ｑｕｅｅｎｓ Ｕｎｉｖｅｒｓｉｔｙ， Ｏｎｔａｒｉｏ， ＣＮのＤｒ． Ｋ． Ｐｏｏｌｅの研究室から得た。トブラマイシンは、Ｗｉｎｔｈｒｏｐ−Ｕｎｉｖｅｒｓｉｔｙ Ｈｏｓｐｉｔａｌ， Ｍｉｎｅｏｌａ， ＮＹのＰｈａｒｍａｃｙ Ｄｅｐａｒｔｍｅｎｔから得た。

ＮＮ＝トブラマイシン；ＢＥ＝ＢｉｓＥＤＴ、０．４μｇ／ｍｌ；菌株は、Ｄｅｐａｒｔｍｅｎｔ ｏｆ Ｐｅｄｉａｔｒｉｃｓ ａｎｄ Ｃｏｍｍｕｎｉｃａｂｌｅ Ｄｉｓｅａｓｅ， Ｕｎｉｖｅｒｓｉｔｙ ｏｆ Ｍｉｃｈｉｇａｎ， Ａｎｎ Ａｒｂｏｒ， ＭＩのＤｒ． Ｊ． Ｊ． ＬｉＰｕｍａの研究室から；そして同じくＶｅｌｏｉｒａ．他，２００３により得た。トブラマイシンは、Ｗｉｎｔｈｒｏｐ−Ｕｎｉｖｅｒｓｉｔｙ Ｈｏｓｐｉｔａｌ， Ｍｉｎｅｏｌａ， ＮＹのＰｈａｒｍａｃｙ Ｄｅｐａｒｔｍｅｎｔから得た。

ＮＮ＝トブラマイシン；ＢＥ＝ＢｉｓＥＤＴ、０．４μｇ／ｍｌ；菌株は、Ｄｅｐａｒｔｍｅｎｔ ｏｆ Ｐｅｄｉａｔｒｉｃｓ ａｎｄ Ｃｏｍｍｕｎｉｃａｂｌｅ Ｄｉｓｅａｓｅ， Ｕｎｉｖｅｒｓｉｔｙ ｏｆ Ｍｉｃｈｉｇａｎ， Ａｎｎ Ａｒｂｏｒ， ＭＩのＤｒ． Ｊ． Ｊ． ＬｉＰｕｍａの研究室から；そして同じくＶｅｌｏｉｒａ．他，２００３により得た。トブラマイシンは、Ｗｉｎｔｈｒｏｐ−Ｕｎｉｖｅｒｓｉｔｙ Ｈｏｓｐｉｔａｌ， Ｍｉｎｅｏｌａ， ＮＹのＰｈａｒｍａｃｙ Ｄｅｐａｒｔｍｅｎｔから得た。

ＮＮ＝トブラマイシン；ＢＥ＝ＢｉｓＥＤＴ、０．８μｇ／ｍｌ；Ｌｉｐｏ−ＢＥ−ＮＮ＝リポソームＢＥ−ＮＮ；菌株は、Ｄｅｐａｒｔｍｅｎｔ ｏｆ Ｃｈｅｍｉｓｔｒｙ ａｎｄ Ｂｉｏｃｈｅｍｉｓｔｒｙ， Ｌａｕｒｅｎｔｉａｎ Ｕｎｉｖｅｒｓｉｔｙ， Ｏｎｔａｒｉｏ， ＣＮのＤｒ． Ａ． Ｏｍｒｉの研究室から得た（Ｍ株は、粘液様Ｂ．セパシア；ＰＡ＝緑膿菌；ＳＡ＝黄色ブドウ球菌）。トブラマイシンは、Ｗｉｎｔｈｒｏｐ−Ｕｎｉｖｅｒｓｉｔｙ Ｈｏｓｐｉｔａｌ， Ｍｉｎｅｏｌａ， ＮＹのＰｈａｒｍａｃｙ Ｄｅｐａｒｔｍｅｎｔから得た。

ＮＮ＝トブラマイシン；ＢＥ＝ＢｉｓＥＤＴ、０．８μｇ／ｍｌ；Ｌｉｐｏ−ＢＥ−ＮＮ＝リポソームＢＥ−ＮＮ；菌株は、Ｄｅｐａｒｔｍｅｎｔ ｏｆ Ｃｈｅｍｉｓｔｒｙ ａｎｄ Ｂｉｏｃｈｅｍｉｓｔｒｙ， Ｌａｕｒｅｎｔｉａｎ Ｕｎｉｖｅｒｓｉｔｙ， Ｏｎｔａｒｉｏ， ＣＮのＤｒ． Ａ． Ｏｍｒｉの研究室から得た（Ｍ株は、粘液様Ｂ．セパシア；ＰＡ＝緑膿菌；ＳＡ＝黄色ブドウ球菌）。トブラマイシンは、Ｗｉｎｔｈｒｏｐ−Ｕｎｉｖｅｒｓｉｔｙ Ｈｏｓｐｉｔａｌ， Ｍｉｎｅｏｌａ， ＮＹのＰｈａｒｍａｃｙ Ｄｅｐａｒｔｍｅｎｔから得た。

ＢＥ＝ＢｉｓＥＤＴ；ＮａＰＹＲ＝ナトリウムピリチオン；化学薬品は、Ｓｉｇｍａ Ａｌｄｒｉｃｈから得た。太字は相乗性。示された菌株は、Ａｍｅｒｉｃａｎ Ｔｙｐｅ Ｃｕｌｔｕｒｅ Ｃｏｌｌｅｃｔｉｏｎ （ＡＴＣＣ， Ｍａｎａｓｓａｓ， ＶＡ）からのものであった。 The results are shown in Tables 2-17.

BE = 0.2 μg / ml BisEDT; The strain was obtained from the Clinical Microbiology Laboratory of Winthrop-University Hospital, Mineola, NY. Nafcillin was obtained from Sigma (St. Louis, MO).

BE = 0.2 μg / ml BisEDT; The strain was obtained from the Clinical Microbiology Laboratory of Winthrop-University Hospital, Mineola, NY. Nafcillin was obtained from Sigma.

BE = 0.2 μg / ml BisEDT; Strain S2446-3 was obtained from Clinical Microbiology Laboratory of Winthrop-University Hospital, Minnea, NY. The antibody was obtained from Sigma.

GM = gentamicin; Strain S2400-1 was obtained from the Clinical Microbiology Laboratory of Wintrop-University Hospital, Mineola, NY. Gentamicin was obtained from Pharmatrop Department of Winthrop. Bold is synergistic.

Data in μg / ml; Strain S2400-1 was obtained from the Clinical Microbiology Laboratory of Wintrop-University Hospital, Mineola, NY. Antibiotics were obtained from Winthrop's Pharmacy Department.

Data in μg / ml; Strain S2400-1 was obtained from the Clinical Microbiology Laboratory of Wintrop-University Hospital, Mineola, NY. Antibiotics were obtained from Winthrop's Pharmacy Department.

Data in μg / ml; Strain S2400-1 was obtained from the Clinical Microbiology Laboratory of Wintrop-University Hospital, Mineola, NY. Antibiotics were obtained from Winthrop's Pharmacy Department.

Data in μg / ml; Antibiotics were obtained from Pharmacy Department of Winthrop-University Hospital, Mineola, NY.

AB = antibiotics; CM = chloramphenicol; AM = ampicillin; BE = 0.3 μg / ml BisEDT, strains are from Department of Molecular Genetics and Cell Biology, The University of Chicago, IL, IL. Obtained from the laboratory of MJ Casadaban. Antibiotics were obtained from Pharmacy Department of Winthrop-University Hospital, Mineola, NY.

DOX = doxycycline; BE = 0.3 μg / ml BisEDT; strains are from Department of Bacteriology, The University of Bristol, Bristol, UK Obtained from I Chopra's laboratory. Antibiotics were obtained from Pharmacy Department of Winthrop-University Hospital, Mineola, NY.

Agr = aminoglycoside resistance; NN = tobramycin; PA = Pseudomonas aeruginosa; BE = BisEDT, 0.3 μg / ml; strains are from Department of Microbiology and Immunology, Queens University, Ontario, K. Obtained from Poole laboratory. Tobramycin was obtained from Pharmacy Department of Winthrop-University Hospital, Mineola, NY.

NN = tobramycin; BE = BisEDT, 0.4 μg / ml; strains are from Dr. of Department of Pediatrics and Communicable Disease, University of Michigan, Ann Arbor, MI. J. et al. J. et al. From LiPuma's laboratory; and also Veloira. Et al., 2003. Tobramycin was obtained from Pharmacy Department of Winthrop-University Hospital, Mineola, NY.

NN = tobramycin; BE = BisEDT, 0.4 μg / ml; strains are from Dr. of Department of Pediatrics and Communicable Disease, University of Michigan, Ann Arbor, MI. J. et al. J. et al. From LiPuma's laboratory; and also Veloira. Et al., 2003. Tobramycin was obtained from Pharmacy Department of Winthrop-University Hospital, Mineola, NY.

NN = tobramycin; BE = BisEDT, 0.8 μg / ml; Lipo-BE-NN = liposome BE-NN; strains are from Department of Chemistry and Biochemistry, Laurentian University, Ontario, N. A. Obtained from Omri's laboratory (M strain is mucus-like B. cepacia; PA = Pseudomonas aeruginosa; SA = S. Aureus). Tobramycin was obtained from Pharmacy Department of Winthrop-University Hospital, Mineola, NY.

NN = tobramycin; BE = BisEDT, 0.8 μg / ml; Lipo-BE-NN = liposome BE-NN; strains are from Department of Chemistry and Biochemistry, Laurentian University, Ontario, N. A. Obtained from Omri's laboratory (M strain is mucus-like B. cepacia; PA = Pseudomonas aeruginosa; SA = S. Aureus). Tobramycin was obtained from Pharmacy Department of Winthrop-University Hospital, Mineola, NY.

BE = BisEDT; NaPYR = sodium pyrithione; chemicals were obtained from Sigma Aldrich. Bold is synergistic. The strains shown were from the American Type Culture Collection (ATCC, Manassas, Va.).

Example 7
Comparison of the effects of bismuth-thiol (BT) and antibiotics against gram-positive and gram-negative bacteria, including antibiotic-resistant strains. Evaluated against multiple clinical isolates of negative bacteria.

Materials and Methods Test compounds and test concentration ranges are as follows: BisEDT (Domenico et al 1997; Domenico et al., Antimicrob. Agents. Chemother. 45 (5): 1414-1421, and Example 1), 16-0.015 μg / mL; linezolid (ChemPacifica Inc., # 35710), 64-0.06 μg / mL; daptomycin (Cubist Pharmaceuticals # MCB2007), 32-0.03 μg / ml and 16-0.015 μg / mL; Vancomycin (Sigma-Aldrich, St. Louis, MO, # V2002), 64-0.06 μg / mL; ceftazidime (Sigma # C3809), 6 4-0.06 μg / mL and 32-0.03 μg / mL; imipenem (US Pharmacopeia, NJ, # 1337809), 16-0.015 μg / mL and 8-0.008 μg / mL; ciprofloxacin (US Pharmacopoeia, # IOC265), 32-0.03 μg / mL and 4-0.004 μg / mL; Gentamicin (Sigma # G3632) 32-0.03 μg / mL and 16-0.015 μg / mL. All test substances except gentamicin were dissolved in DMSO, and gentamicin was dissolved in water. Stock solutions were prepared at 40 times the highest concentration in the test plate. The final concentration of DMSO in the inspection system was 2.5%.

Organisms Examination organisms were obtained from the following clinical laboratories: CHP, Clarian Health Partners, Indianapolis, IN; Center, Public Health Institute Tuberculosis Center, New York, NY; ATCC, American Type Culture Collection, Manassas, VA; Mt Sinais Hp. , Mount Sinai Hospital, New York, NY; UCSF, University of California San Francisco General Hospital, San Francisco, CA; Bronson Hospital, Bronson Methodist Hospital, Kalamazoo, MI; isolates for quality management, American Type Culture Collection (ATCC , Manassas, VA). The organisms were streaked for isolation on an agar medium appropriate for each organism. Colonies were removed from the isolation plate by swab and placed in suspension in an appropriate broth containing cryoprotectant. The suspension was aliquoted into cold storage vials and kept at -80 ° C. Abbreviations: BisEDT, bismuth-1,2-ethanedithiol; LZD, linezolid; DAP, daptomycin; VA, vancomycin; CAZ, ceftazidime; IPM, imipenem; CIP, ciprofloxacin; GM, gentamicin; MSSA, methicillin sensitive staphylo CLSI QC, quality control strain of Clinical and Laboratory Standards Institute; MRSA, methicillin-resistant Staphylococcus aureus; CA-MRSA, community-acquired methicillin-resistant Staphylococcus aureus; MSSE, methicillin susceptibility Epidermidis; MRSE, methicillin-resistant Staphylococcus epidermidis; VSE, vancomycin sensitivity Enterococcus.

  Isolates can be obtained from frozen vials in appropriate media (Trypticase Soy Agar (Becton-Dickinson, Sparks, MD) for most organisms, or Trypticase Soy Agar for Streptococcus + 5% sheep blood (Clevelandfic). Streaked up. Plates were incubated overnight at 35 ° C. Quality control organisms were included. The medium used for the MIC assay was Mueller Hinton II Broth (MHB II-Becton Dickinson, # 212322) for most organisms. MHBII was supplemented with 2% dissolved horse blood (Cleveland Scientific Lot # H13913) and adapted to the development of Streptococcus pyogenes and Streptococcus agalactia. The medium was prepared at 102.5% of normal weight to offset the dilution caused by adding 5 μL of drug solution to each well of the microdilution panel. In addition, testing with daptomycin supplemented the medium with an additional 25 mg / L Ca 2+.

  MIC assay, Clinical and Laboratory Standards Institute (Clinical and Laboratory Standards Institute Methods for Dilution Antimicrobial Susceptibyity Tests for Bacteria That Grow Aerobically;.. Approved Standard-Seventh Edition Clinical and Laboratory Standards Institute document M7-A7 [ISBN 1-56238- 587-9] Clinical and Laboratory Standards Institute Ute, 940 West Valley Road, Suite 1400, Wayne, Pennsylvania 19087-1898 USA, 2006), and serial dilution and liquid transfer were performed using an automatic liquid handler. Automatic liquid handlers included Multidrop 384 (Labsystems, Helsinki, Finland), Biomek 2000 and Multimek 96 (Beckman Coulter, Fullerton CA). The wells in rows 2-12 of a standard 96 well microdilution plate (Falcon 3918) were filled with Multidrop 384 with 150 μL of water for DMSO or gentamicin. Drug (300 μL) was dispensed into the first column of the appropriate row in these plates. These became mother plates when preparing inspection plates (daughter plates). Biomek 2000 completed the series movement up to the eleventh row of the mother plate. The wells in the twelfth row contained no drug and were organism growth control wells on the daughter plate. 185 μL of the appropriate test medium (described above) was added to the daughter plate using a Multidrop 384. Daughter plates were prepared in a Multimek 96 instrument by transferring 5 μL of the drug solution from each well of the mother plate to each corresponding well of each daughter plate in one step.

  A standard inoculum for each organism was prepared by the CLSI method (ISBN 1-56238-587-9, cited above). A suspension was prepared in MHB to equal the turbidity of a 0.5 McFarland standard. The suspension was diluted 1: 9 with broth appropriate for the organism. The inoculum of each organism was dispensed into vertically separated sterile reservoirs (Beckman Coulter) and inoculated into plates using Biomek 2000. The daughter plate was placed on the working surface of the Biomek 2000, which was reversed so that inoculation was performed from a low drug concentration to a high drug concentration. Biomek 2000 delivered 10 μL of the standardized inoculum to each well. This resulted in a final cell concentration of the daughter plate of approximately 5 × 10 5 colony forming units / mL. The daughter plate wells thus finally contained 185 μL broth, 5 μL drug solution, and 10 μL bacterial inoculum. Plates were stacked in triplicate, the top plate was covered, placed in a plastic bag, and most of the isolates were incubated at 37 ° C. for approximately 18 hours. Streptococcus plates were read after 20 hours of incubation. The microplate was viewed from the bottom using a plate viewer. In each of the test media, an uninoculated soluble control plate was observed for evidence of drug precipitation. The MIC was read and recorded as the lowest drug concentration that inhibited the visual development of the organism.

Results All commercially available drugs were soluble at all test concentrations in both media. BisEDT showed a small amount of precipitate at 32 μg / mL but did not affect the MIC reading because the inhibitory concentration of all tested organisms was well below that concentration. On each assay day, the appropriate quality control strain was included in the MIC assay. MIC values obtained from these strains, as appropriate, published for each drug (Clinical and Laboratory Standards Institute-Performance Standards for Bioscience Sustainable Stift. 18-National Institute of Biomedical Sustainability. -653-0] Clinical and Laboratory Standards Institute, 940 West Valley Road, Suite 1400, Wayne, Pennsylvania 19087-1898 USA, 2008).

  On each assay day, the appropriate quality control strain was included in the MIC assay. MIC values obtained from these strains, as appropriate, published for each drug (Clinical and Laboratory Standards Institute. -653-0]). Of the 141 types of quality control strains for which the quality control range was announced, 140 types (99.3%) were within the specified range. One exception was imipenem against S. aureus 29213, where one figure obtained in a single run was one dilution lower than the published QC range (0.008 μg / mL or less). All other quality control results during the implementation were within the specified quality control scope.

  BisEDT demonstrated potent activity against methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant Staphylococcus aureus (MRSA), and community-acquired MRSA (CA-MRSA) and was tested below 1 μg / mL All strains were inhibited, and the MIC90 value was 0.5 μg / mL for all three organism groups. BisEDT showed greater activity than linezolid and vancomycin, and equivalent activity to daptomycin. Imipenem was more potent than BisEDT against MSSA (MIC90 = 0.03 μg / mL). However, MRSA and CAMRSA were resistant to imipenem, while BisEDT demonstrated activity comparable to that shown for MSSA. BisEDT was highly active against methicillin sensitive and methicillin resistant Staphylococcus epidermidis (MSSE and MRSE) with MIC90 of 0.12 and 025 μg / mL, respectively. BisEDT was more active against MSSE than any of the other drugs tested other than imipenem. BisEDT was the most active drug tested against MRSE.

  BisEDT demonstrated activity comparable to daptomycin, vancomycin, and imipenem against vancomycin-sensitive Enterococcus faecalis (VSEfc) with a MIC90 value of 2 μg / mL. Importantly, BisEDT is the most active drug tested against vancomycin-resistant Enterococcus faecalis (VREfc) with a MIC90 value of 1 μg / mL.

  BisEDT was very active against vancomycin-sensitive Enterococcus faecium (VSEfm) with a MIC90 value of 2 μg / mL, which was similar or similar to daptomycin and was one dilution higher than vancomycin . BisEDT and linezolid were the most active drugs tested against vancomycin-resistant Enterococcus faecium (VREfm), each demonstrating a MIC90 value of 2 μg / mL. The activity of BisEDT against Streptococcus pyogenes (MIC90 value of 0.5 μg / mL) was comparable to vancomycin, greater than linezolid, and slightly lower than daptomycin and ceftazidime. The compound inhibited all strains tested at 0.5 μg / mL or less. In these studies, the bacterial species that is at least sensitive to BisEDT was Streptococcus agalactia, and the observed MIC90 value was 16 μg / mL. BisEDT was less active than all drugs tested except for gentamicin.

  The activity of BisEDT and comparative substances against gram-negative bacteria included demonstrated the efficacy of BisEDT against Acinetobacter baumannii (MIC 90 value 2 μg / mL), making BisEDT the most active compound tested. A significant number of test isolates for comparative agents have increased MICs, and these drugs have MIC90 values outside the measurement range. BisEDT was a potent inhibitor of E. coli and inhibited all strains at 2 μg / mL or less (MIC90 = 2 μg / mL). The compound was less active than imipenem, but more active than ceftazidime, ciprofloxacin and gentamicin. BisEDT also demonstrated activity against Klebsiella pneumoniae, with a MIC90 value of 8 μg / mL, equivalent to imipenem. The relatively high MIC90 values exhibited by imipenem, ceftazidime, ciprofloxacin, and gentamicin indicated that this is a group of organisms with high antibiotic resistance. BisEDT is the most active compound tested against P. aeruginosa with a MIC90 value of 4 μg / mL. There was a high level of resistance to the comparator in this group of test isolates.

  In summary, BisEDT displays a broad spectrum capability for multiple clinical isolates representing multiple species, including those commonly involved in acute and chronic skin and skin structure infections in humans. The activities of BisEDT and important comparators were evaluated against 723 clinical isolates of gram positive and gram negative bacteria. BT compounds demonstrated broad spectrum activity, and for a number of test organisms in this study, BisEDT was the most active compound tested for antimicrobial activity. BisEDT is available from MSSA, MRSA, CA-MRSA, MSSE, MRSE and S.M. It was most active against Piogenes, and the MIC90 value was 0.5 μg / mL or less. Strong activity is observed in VSEfc, VREfc, VSEfm, VREfm, A. Also demonstrated in Baumany, E. coli, and Pseudomonas aeruginosa, MIC90 values were in the range of 1-4 μg / mL. The MIC90 value is Pneumoniae (MIC90 = 8 μg / mL) and S. pneumoniae. Observed with agalactia (MIC90 = 16 μg / mL).

Example 8
Enhanced and synergistic activity of particulate BT-antibiotics This example shows that particulate bismuth-thiol (BT) promotes antibiotic activity through potentiating and / or synergistic interactions.

  A major complication in treating infection is the emergence of bacterial antibiotic resistance. S. Methicillin resistance in Epidermidis (MRSE) and Staphylococcus aureus (MRSA) actually represents multidrug resistance, which makes it very difficult to eradicate these pathogens. However, hundreds of strains of Staphylococcus tested did not show resistance to BT. Furthermore, sub-inhibition (subMIC) concentrations of BT reduced resistance to several important antibiotics.

  Providing a graphical representation of the antibiotic-resensitizing effects of bismuth ethanedithiol (BisEDT) of subMIC against Staphylococcus aureus MRSA (FIG. 4), gentamicin, cefazolin, cefepime, imipenem, sulfa It shows potentiating antibiotic action with several classes of antibiotics including methoxazole and levofloxacin. That is, BisEDT nonspecifically enhanced the activity of most antibiotics.

１２のＭＲＳＡ臨床単離物をＢＨＩＧ／Ｘ中で発育させて、０〜０．１μｇ／ｍＬ ＢｉｓＥＤＴの存在下で抗生物質の系列希釈物に暴露した。μｇ／ｍＬで計算されたＭＩＣおよびＢＰＣは、少なくとも３回のトライアルから得た平均±標準偏差である。右側の列は、抗生物質感受性（Ｓ）および耐性（Ｒ）に関する標準ＭＩＣを列挙している。 Antimicrobial susceptibility testing by broth dilution was performed on 12 MRSA strains using multiple antibiotics in combination with subMIC levels of BisEDT. Both biofilm preventive concentration (BPC) and minimum inhibitory concentration (MIC) were measured with a special biofilm medium (BHIG / X). The MIC and BPC of gentamicin and cefazolin were reduced by subMIC BisEDT (BisEDT MIC, 0.2-0.4 μg / mL), but not below the sensitive breakpoint. The subMIC BisEDT enhanced the sensitivity of MRSA to gatifloxacin and cefepime to near the breakpoint of sensitivity. These strains were already sensitive to vancomycin, but became more pronounced in the presence of subMIC BisEDT. In general, MIC and BPC were reduced 2-5 fold by subMIC BisEDT.

Twelve MRSA clinical isolates were grown in BHIG / X and exposed to serial dilutions of antibiotics in the presence of 0-0.1 μg / mL BisEDT. The MIC and BPC calculated in μg / mL are the mean ± standard deviation obtained from at least 3 trials. The right column lists standard MICs for antibiotic susceptibility (S) and resistance (R).

１２種のセフェピム耐性ＭＲＳＡを、ｓｕｂＭＩＣのＢｉｓＥＤＴと組み合わせたセフェピムへの感受性について、ポリスチレンプレート内のＢＨＩＧ／Ｘ培地で３７℃で４８時間検査した。 The broth dilution test of cefepime resistant MRSA isolates is shown in Table 19. 0.1 μg / mL BisEDT significantly enhanced the inhibitory activity of cefepime in 11 out of 12 isolates. In this particular study, the data showed synergy between BisEDT and cefepime (FIC <0.5), with many of the isolates at sensitive breakpoints.

Twelve cefepime resistant MRSA were tested for 48 hours at 37 ° C. in BHIG / X medium in polystyrene plates for sensitivity to cefepime in combination with subMIC BisEDT.

ＮＡＦまたはＧＭはμｇ／ｍＬを表わし；ＢＥは０．２μｇ／ｍＬ The results of the combination test with nafcillin or gentamicin are shown in Table 20. BisEDT (0.2 μg / mL) in combination with nafcillin reduced nafcillin MIC90 more than 4-fold relative to MRSA (FIC, 0.74). BisEDT in combination with gentamicin reduced gentamicin's MIC90 more than 10-fold over MRSA (FIC, 0.6). BT suppressed the resistance of all four of the gentamicin resistant isolates tested to clinically relevant concentrations [Domenico et al. , 2002]. The MIC of these antimicrobial agents was substantially reduced, especially with gentamicin. The broth used in these studies was Trypticase Soy Broth (TSB) containing 2% glucose, showing similar results to those observed in Mueller-Hinton II broth supplemented with 1% sheep blood.

NAF or GM represents μg / mL; BE is 0.2 μg / mL

  Staphylococcus epidermidis The activity of most classes of antibiotics was enhanced in the presence of BisEDT. For BPC, clindamycin and gatifloxacin are S. cerevisiae when combined with BisEDT. It showed significantly higher anti-biofilm activity against epidermidis (FIG. 5). Stated differently, clindamycin, gatifloxacin and gentamicin BPC were reduced by 50-fold, 10-fold and 4-fold in the presence of subMIC BisEDT, respectively.

  A slight reduction in biofilm preventive concentration (BPC) was observed with minocycline, vancomycin, and cefazolin, but rifampicin and nafcillin still showed no effect at 0.05 μg / mL BisEDT. With 0.1 μg / mL BisEDT, no biofilm was detected regardless of the antibiotic used, indicating that no antagonism occurred. This BisEDT concentration is determined by S. cerevisiae. It approximated the epidermidis MIC [Domenico et al. , 2003] (see FIG. 5).

  Of the 8 antibiotics tested for growth inhibition, 7 It was significantly enhanced against epidermidis in the presence of 0.1 μg / mL (0.5 μM) BisEDT (FIG. 6). The MIC variation was most pronounced with clindamycin and gentamicin, followed by vancomycin, cefazolin, minocycline, gatifloxacin and nafcillin, and rifampicin had no effect. These strains were resistant to (NC, CZ, GM, CM) of antibiotics, and only cefazolin resistance was suppressed to clinically relevant levels by BisEDT.

  S. The minimum bactericidal concentration (MBC) of most antibiotics tested against epidermidis was slightly reduced by subMIC BisEDT. Gentamicin shows the greatest reduction in MBC (4- to 16-fold), followed by cefazolin (4- to 5-fold), vancomycin and nafcillin (3- to 4-fold), minocycline and gatifloxacin (2- to 3-fold) ) And clindamycin and rifampicin MBC were still not significantly affected. It is explained that clindamycin is a bacteriostatic agent and has no bactericidal activity. Cefazolin resistance was suppressed for MBC [Domenico et al. , 2003]. These effects were additive.

  Antimicrobial efficacy has also been demonstrated in an in vivo rat graft infection model (Table 21). With a low level of BisEDT of 0.1 μg / mL, 7 days resistance The prevention of epidermidis biofilm could be promoted.

As summarized in Table 21, implants impregnated with 0.1 μg / mL BisEDT, 10 μg / mL RIP and 10 μg / mL rifampin alone or in combination were implanted subcutaneously into rats. A physiological solution (1 ml) containing 2 × 10 7 cfu / ml of MS and MR strains was inoculated on the surface of the graft using a tuberculin syringe. All grafts were removed 7 days after transplantation and sonicated in sterile saline for 5 minutes to remove attached bacteria. Quantification of viable bacteria was obtained by culturing the dilutions on blood agar plates. The detection limit was approximately 10 cfu / cm2.

緑膿菌の耐性株を、トブラマイシン（ＮＮ）およびＢｉｓＥＤＴ（ＢＥ；０．３３μｇ／ｍｌ）の存在下、３７℃のＭｕｅｌｌｅｒ−Ｈｉｎｔｏｎ ＩＩブロス中で培養した。ＭＩＣを、２４±１時間の発育を阻害する抗生物質濃度として測定した。 The activity of tobramycin against gram-negative bacteria-resistant Pseudomonas aeruginosa was enhanced several times by subMIC BisEDT (Table 22). In these trials, the MIC was more accurately defined as IC24.

A resistant strain of Pseudomonas aeruginosa was cultured in Mueller-Hinton II broth at 37 ° C. in the presence of tobramycin (NN) and BisEDT (BE; 0.33 μg / ml). MIC was measured as the concentration of antibiotic that inhibits growth for 24 ± 1 hour.

In contrast to tobramycin-resistant Burkholderia cepacia, 0.4 μg / mL BisEDT made 7 of 10 isolates tobramycin sensitive (mean FIC; 0.48) and reduced MIC90 10-fold ( Table 23). Both tobramycin MIC and MBC were significantly reduced by subMIC BisEDT to a level that was feasible for 50 clinical Burkholderia cepacia isolates [Veloria et al. , 2003]. Liposomal forms of BisEDT and tobramycin have a high synergy demonstrated against Pseudomonas aeruginosa (Halwani et al., 2008; Halwani et al., 2009).

大腸菌の耐性株を、単独または組み合わせでのクロラムフェニコール（ＣＭ）またはアンピシリン（ＡＭＰ）およびＢｉｓＥＤＴの存在下、３７℃のＭｕｅｌｌｅｒ−Ｈｉｎｔｏｎ ＩＩブロス中で培養した（ＢＥ：０．３３μｇ／ｍｌ）。ＭＩＣを、２４±１時間の発育を阻害した抗生物質濃度として測定した。 Chloramphenicol and ampicillin resistant E. coli were made sensitive to these drugs with the addition of subMIC BisEDT (Table 24).

Resistant strains of E. coli were cultured in Mueller-Hinton II broth at 37 ° C. in the presence of chloramphenicol (CM) or ampicillin (AMP) and BisEDT, alone or in combination (BE: 0.33 μg / ml) . MIC was measured as the concentration of antibiotic that inhibited growth for 24 ± 1 hour.

大腸菌の耐性株を、単独または組み合わせでのドキシサイクリン（ＤＯＸ）およびＢｉｓＥＤＴの存在下、３７℃のＭｕｅｌｌｅｒ−Ｈｉｎｔｏｎ ＩＩブロス中で培養した（ＢＥ：０．３３μｇ／ｍｌ）。ＭＩＣを、２４±１時間の発育を阻害した抗生物質濃度として測定した。 Tetracycline-resistant E. coli became sensitive to tetracycline upon addition of subMIC BisEDT (Table 25). The combination was synergistic for TET M and TET D strains (FIC ≦ 0.5) and had an additive effect on TET A and TET B strains.

Resistant strains of E. coli were cultured in Mueller-Hinton II broth at 37 ° C. (BE: 0.33 μg / ml) in the presence of doxycycline (DOX) and BisEDT alone or in combination. MIC was measured as the concentration of antibiotic that inhibited growth for 24 ± 1 hour.

References Domenico P, R O'Leary, BA Cunha. 1992. Differential effect of bismuth and salicylate compounds on antibiotic sensitivity of Pseudomanas aeruginosa. Eur J Clin Microbiol Infect Dis 11: 170-175; Domenico P, D Parikh, BA Cunha. 1994. Bismuth modulation of antibacterial activity, against gastrointestinal bacterial pathogens. Med Microbiol Lett 3: 114-119; Domenico P, Kazzaz JA, Davis JM, Niederman MS. 2002. Subinhibit bimusmu etherethiol (BisEDT) sensitives resist Staphylococcus aureus to nafcillin or gentamicin. Annual Meeting, ASM, Salt Lake City, UT; Domenico P, Kazzaz JA, Davis JM. 2003. Combining antibiotic resistance with bismuth-thiol. Research Advances in Antimicrob Agents Chemother 3: 79-85; 2004. BisEDT and RIP act in synergy to prevent graft effects by resist staphylococci. Peptides 25: 2047-2053; Halwani M, Bloome S, Suntres ZE, Alipore M, Azghani AO, Kumar A, Omri A. 2008. Liposomal bismuth-ethanediolformation enhancements antimicrobial activity of tobramcin. Intl J Pharmaceut 358: 278-84; Halwani M, Hebert S, Suntres ZE, Lafrénie RM, Azghani AO, Omri A. et al. 2009. Bismuth-thiol enhancement enhancements bioactives of liposomal tobramcincin aginest biofilm and quorum sensation moleculare. Int J Pharmaceut 373: 141-6; Veloira WG, Gurzenda EM, Domenico P, Davis JM, Kazzaz JA. 2003. Synergy of tobramycin and bismuth thiols against Burkholderia cepacia. J Antimicrob Chemother 52: 915-919.

ＳＡ、スタフィロコッカス・アウレウス；ＭＲＳＡ、メチシリン耐性スタフィロコッカス・アウレウス；ＥＦｃ、エンテロコッカス・フェカーリス；ＳＰ、ストレプトコッカス・ニューモニエ；ＰＲＳＰ、ペニシリン耐性ストレプトコッカス・ニューモニエ；ＥＣ、大腸菌；ＫＰ、クレブシエラ・ニューモニエ；ＰＡ、シュードモナス・エルギノーサ；Ｂｃｅｐ、バークホルデリア・セパシア；Ｂｍｕｌｔ、バークホルデリア・マルチボランス；Ａｂａｕ、アシネトバクター・バウマニー；Ｍｓｍｅｇ、マイコバクテリウム・スメグマチス Example 9
Particulate BT-Antibiotic Enhancement and Synergistic Activity This example shows that BisEDT, a particulate bismuth thiol, has antibiotic activity through enhanced and / or synergistic interactions with specific antibiotics against specific microbial target organisms. To promote. One-point data for each combination shown in Table 26 was created essentially according to the method used in Example 8.

SA, Staphylococcus aureus; MRSA, methicillin-resistant Staphylococcus aureus; EFc, Enterococcus faecalis; SP, Streptococcus pneumoniae; PRSP, penicillin-resistant Streptococcus pneumoniae; EC, E. coli; KP, Klebsiella pneumoniae; Pseudomonas aeruginosa; Bcep, Burkholderia cepacia; Bmult, Burkholderia multiborance; Abau, Acinetobacter baumannii; Msmeg, Mycobacterium smegmatis

Example 10
Enhanced and synergistic activity of microparticulate BT-antibiotics To test the effect of a combination of microparticulate BisEDT and four BisEDT analogs prepared as described above with other agents on representative strains of multiple Gram-negative pathogens did. Utilizing a variation of the general test method, the partial inhibitory concentration (FIC) and the FIC index (FICI) used were synergistic (FICI ≦ 0.5), enhancing (0.5 <FICI ≦ 1.0) ), Antagonistic (FICI> 4.0) and insignificant (1.0 <FICI ≦ 4.0) were determined (Antibiotics in Laboratory Medicine, edited by V Lorian. Williams and Wikins, Baltimore, MD). Edition, pp. 432-492, Elipoulos G and R Moellering., 1991. Antimicrobial combinations .; Odds, 2003 J. Antimicrob.Chemother.52 (1): 1). The checkerboard technique was used to determine the FIC index and used for this test.

  Stock solutions of all test substances were prepared 40 times the final target concentration with an appropriate solvent. All test substances were in solution under these conditions. Unless the strain is totally resistant to the test agent, the final drug concentration in the FIC assay plate was set so that the MIC values of each drug for each test organism were collected. The tested density range is shown in Table 27. Test organisms were initially obtained from clinical sources or from the American Type Culture Collection. Upon acquisition, the isolate was streaked on Tryptic Soy Agar II (TSA). Colonies were collected from these plates and cell suspensions were prepared in an appropriate broth growth medium containing a cryoprotectant. Thereafter, aliquots were frozen at -80 ° C. Frozen seeds of organisms to be tested in a given assay were thawed and streaked onto TSA plates for incubation and incubated at 35 ° C. All organisms were examined with Mueller-Hinton II Broth (Beckton Dickinson, Lot No. 9044411). Broth was prepared at 1.05 times the normal weight / volume to offset the 5% volume of drug in the final test plate.

  Final inhibitory concentration (MIC) values were measured as before using the Broth microdilution method for aerobic bacteria (Clinical and Laboratory Standards Institute (CLSI). Applied Standard-Eight Edition. CLSI document M07-A8 [ISBN 1-56238-689-1] .Clinical and Laboratory Standards Institute, 940 West Valley Valley, 1940. e, Pennsylvania 19087-1898 USA, 2009).

  FIC values were measured using the broth microdilution method described previously (Sweeney et al., 2003 Antimicrob. Agents Chemother. 47 (6): 1902-1906). To prepare test plates, serial liquid dilution and liquid transfer were performed using an automated liquid handler (Multidrop 384, Labsystems, Helsinki, Finland; Biomek 2000 and Multimek 96, Beckman Coulter, Fullerton CA).

  Using Multidrop 384, 150 μL of the appropriate solvent was loaded into the second through twelfth rows in the appropriate wells of a standard 96 well microdilution plate (Falcon 3918). 300 μL of each second test agent was added to each well in the first row of the plate. These plates were used to prepare drug “mother plates” that provide serial dilutions for drug combination plates. Using Biomek 2000, 150 μL of each second drug solution (40 ×) was transferred from the first row of wells of the mother plate to generate 11 2 × dilution series. Bis-EDT (and analog) mother plates were serially diluted from top to bottom using a multichannel pipette. Combine two mother plates, one for each second drug and one for Bis-EDT (or analog), and transfer (using a multichannel pipette) the same volume to the drug combination plate As a result, a “checkerboard” pattern was formed. Row H and column 12 each contained serial dilutions of one of the drugs for which the MIC was measured.

  180 μL of test medium was added to the “daughter plate” using Multidrop 384. Thereafter, using Multimek 96, 10 μL of the drug solution was transferred from each well of the drug combination mother plate to each corresponding well of the daughter plate in one step. Finally, the test organism was inoculated on the daughter plate. A standardized inoculum for each organism was prepared according to published guidelines (CLSI, 2009). For all isolates, the inoculum of each organism was dispensed into a vertically-sterilized sterile reservoir (Beckman Coulter) and inoculated into the plate using a Biomek 2000. The instrument delivered 10 μL of the standardized inoculum to each well to give a final cell concentration in the daughter plate of approximately 5 × 10 5 colony forming units / mL.

  The test format produced an 8 × 12 checkerboard that tested each compound alone (columns 12 and 8) and combined with drug concentrations in various ratios. All organism plates were stacked in triplicate, the top plate was covered, placed in a plastic bag and incubated at 35 ° C. for approximately 20 hours. After incubation, the microplate was removed from the incubator and viewed from the bottom using a ScienceWare plate viewer. The prepared reading sheet was filled with the MIC of Drug 1 (Row H), the MIC of Drug 2 (Column 12) and the well with and without growth.

  Using the Excel program, FIC was measured by the formula: (MIC of Compound 1 in combination / MIC of Compound 1 alone) + (MIC of Compound 2 in combination / MIC of Compound 2 alone). The FICI for the checkerboard was calculated from each FIC by the formula: (FIC1 + FIC2 + ... FICn) / n, where n = number of each well per plate on which the FIC is calculated. If a single drug produced an MIC result outside the measurement range, the highest next concentration was used as the MIC value in the FIC calculation.

All of the microparticle Bis-EDT, the four microparticle BT analogs, and other drugs (and drug combinations) were soluble at all final test concentrations. The measured MIC and FICI values are shown in the table below.

Example 11
The effect of bismuth-thiol on infection in critical femurs of the rat rat Current care standard methods for open fractures are irrigation, necrotic tissue removal and antibiotics, which do not cause infection of bacterial load in wound It is intended to reduce to a certain extent. Despite these treatments, infections complicate up to 75% of severe open tibial fractures due to war. Interestingly, even though early infections are often caused by Gram-negative bacteria, late infections that are involved in healing problems and cleavage are Gram-positive infections, mostly due to Staphylococcus species (Johnson 2007).

  One reason why S. aureus is resistant to standard treatment is its ability to form biofilms. Bacteria in the biofilm can withstand concentrations of antimicrobial compounds that kill similar organisms in the medium (Costerton 1987).

  The purpose of this study was to determine whether BT alone or with antibiotics would reduce infection in a contaminated open fracture model. The contaminated rat femoral critical defect model is a well-accepted model and was used in the experiment described in this example. This model is a standardized model for comparing various potential treatments and their effects in terms of reducing infection and / or improving healing.

  Although the compounds (CPD) CPD-8-2 (bismuth-pyrithione / butanedithiol; Table 1) and CPD-11 (bismuth pyrithione / ethanedithiol; Table 1) have a different spectrum of activity than Bis-EDT. Rather, it is two BIS-Bis analogs that have shown the ability to biofilm secreting bacteria in vitro.

  Three BT formulations: Bis-EDT, CPD-11 and CPD-8-2 (see Table 1) with or without tobramycin and vancomycin in a polymethylmethacrylate (PMMA) cement bead vehicle. When used, it showed an inhibitory effect against S. aureus strains in vitro. Three formulations of particulate BT were produced in the clinically useful hydrogel form described herein. These BT were tested in suspension in the gel at a concentration of 5 mg / ml-1 found to be the appropriate concentration for gel delivery. The gel formulation conformed to the wound profile and did not need to be removed after application.

  Two treatment objectives were utilized, with BT used alone for the first objective and BT with systemic antibiotics (ABx) for the second objective.

(A) BT alone At 6 hours after inoculation with Staphylococcus aureus, the necrotic tissue of the wound was removed and irrigated with physiological saline, and 1 ml of BT gel was inserted into the defect.

(B) BT and systemic antibiotics (ABx)
Six hours after inoculation with Staphylococcus aureus, debridement of the wound was removed, and irrigated with physiological saline, and 1 ml of the added BT gel was inserted into the defect. The antibiotic used was a dose of cefazolin equal to 5 mg kg-1 delivered by subcutaneous injection twice daily for a total of 3 days after injury. The first dose was administered immediately before necrotic tissue removal. Past data suggested that this dose could reduce bacterial levels from about 106 to about 104, and therefore measure the relative effects of different BT.

(C) Control Six hours after inoculation with Staphylococcus aureus, the necrotic tissue of the wound was removed and ligated with physiological saline. Control animals were also treated with cefazolin with the regimen described previously.

procedure:
In vivo rat injury model procedures were performed as described by Chen et al. (2002 J. Orthop. Res. 20: 142; 2005 J. Orthop. Res. 23: 816; 2006 J. Bone Joint Surg. Am. 88. : 1510; 2007 J. Orthop. Trauma 21: 693). Rats were anesthetized and prepared for surgery. The anterolateral surface of the femoral shaft was exposed through a 3 cm incision. The periosteum and attached muscle were detached from the bone. A polyacetyl plate (27 × 4 × 4 mm) was placed on the anterior-lateral surface of the femur. The plate was pre-drilled and passed through a twisted Kirschner wire of 0.9 mm diameter. These plate substrates were shaped to fit the contours of the femoral shaft. A pilot hole was cut through both femoral cortical bones using a plate as a mold and a twisted Kirschner wire was inserted through the plate and femur. Indentations present at 6 mm intervals on the plate served as a guide for bone removal. In an attempt to prevent thermal damage, the defect was created using a small oscillating saw while cooling the tissue by continuous irrigation.

  Groups of 10 animals were each inoculated with 1 × 10 5 CFU of S. aureus and treated with BT alone or in combination with antibiotics at 6 hours after inoculation as described above. The groups were as follows: Bis-EDT gel; MB-8-2 gel; Bis-EDT gel; Bis-EDT gel &Abx; MB-11 gel &Abx; MB-8-2 gel &Abx; Control (Abx only) ).

  The animals are euthanized 14 days after surgery and the bones and devices are sent for microbial analysis and the results are shown in FIG.

  Based on power analysis, 10 animals per group will give 80% power to detect a 25% difference between the treatment and control groups. In this case, the predicted standard deviation is 35% and α is 0.05.

As shown in FIG. 7, when combined with Bis-EDT, MB-11, and MB-8-2, cefazolin's antibiotic activity is enhanced compared to cefazolin or any Bis compound alone, resulting in yellowness of damaged bone Reduced staphylococcal infection. Cefazolin in combination with MB-11 and MB-8-2 showed high antibiotic activity compared to single cefazolin, indicating reduced S. aureus infection detected on the device. Bis-EDT was not thought to affect cefazolin activity in this capacity.
Example 12
Activity of bismuth-containing compounds against marine organisms

実施例１３
フジツボ着生行動に及ぼすビスマス含有化合物の作用 This example describes the antimicrobial activity of bismuth-containing compounds. MIC values of three bismuth-containing compounds against three different marine bacteria, namely bismuth dimercaprol (BisBAL), bismuth dimercaptotoluene (BisTOL) and bismuth ethanedithiol (BisEDT) are routinely performed by those skilled in the art. The method is confirmed. The data is shown in the table below.

Example 13
Effects of bismuth-containing compounds on barnacle settlement behavior

Compounds BisBAL and BisTOL were included in the assay to determine the inhibitory activity of each compound on barnacle larvae engraftment behavior. The method was performed according to techniques practiced in the art. BisBAL had an EC50 of 1.6 ppm (concentration at which 50% inhibition of growth occurred), and BisTOL had an EC50 of 15.4 ppm. In another experiment, BisEDT was dissolved directly in natural seawater or first dissolved in DMSO and then diluted in natural seawater. EC50 measurements were not statistically different. BisEDT had an EC50 of 1.5 ppm when dissolved directly in seawater and an EC50 of 2.1 ppm when first dissolved in DMSO. The EC50 of the commercially available biocide SEANINE 211 was 0.5 ppm.
Example 14
Effects of bismuth-containing compounds on algae growth

The action of three bismuth-containing compounds on algal growth, namely bismuth dimercaprol (BisBAL), bismuth dimercaptotoluene (BisTOL) and bismuth ethanedithiol (BisEDT), in particular the suppression of germination of Aonori spores. Confirmed ability. Each compound was tested at 0.001, 0.01, 0.1, 1.0 and 10.0 μg / ml. BisEDT was the most effective compound; 1 μg / ml BisEDT inhibited germination of about 50% of the algal spore population and germination of about 75% algal spores at 10 μg / ml. Up to 10 micrograms / ml BisBAL and BisTOL had no inhibitory effect on germination of spores of this particular phase species.
Example 15
Effects of bismuth-containing compounds on algae growth

  Techniques implemented in the art to effect the action of three bismuth-containing compounds on the growth of marine diatoms, namely bismuth dimercaprol (BisBAL), bismuth dimercaptotoluene (BisTOL) and bismuth ethanedithiol (BisEDT) According to By increasing the concentration of the three compounds (0.001, 0.01, 0.1, 1.0, and 10.0 μg / ml), marine diatom epiphytes (diatoms / pits) were suppressed. . Each compound showed inhibitory activity at 0.1 μg / ml; BisEDT was the most active and showed almost 100% inhibition. BisTOL and BisBAL each showed about 30% marine diatom growth at 0.1 μg / ml.

Reference: Costerton et al. , Ann Rev Microbiol. 1987; 41: 435-64; Domenico et al. , Antimicrob Agents and Chemother. 2001; 45 (5): 1417-21; Halwani et al. , IntJ Pharm. 2008; 358: 278-84; Johnson et al. , Clin Infect Dis. 2007; 45 (4): 409-415. ADA Council on Scientific Affairs. Direct and indirect restorative materials. JADA 2003; 134: 463-72. Alliance for Coastal Technologies (ACT). 2004. Bioforming Prevention Technologies for Coastal Sensors / Sensor Platforms. University of Maryland Center of Environmental Science, Workshop Proceedings, November 2003. UMCES Technical Report Series No. TS-426-04-CBL, Solomons, MD. Athanassidias et al. , Aust Dent J 2007; 52: S64-82. Alt et al. , Antimicrob Agents Chemother 2004; 48: 4084-88. Bayston et al. , Biomaterials 2009; 30: 3167-73. Bernardo et al. JADA 2007; 138: 775-783. Beytha et al. , J Dent 2007; 35: 201-206. Bohneretal. J Pharm Sci 1997; 86: 565-72. Bruxton, Eng News 1908; 59, 525; Chem. Abs. , 2: 2010. Bueno et al. Oral Surg Oral Med Oral Path Oral Radio Endo 2009; 107: e65-9. Cao et al. / ACS Applied Materials & Interfaces, 2009; 1: 494. Centers for Disease Control and Prevention (US). Guidelines for infection control in dental health-care settings-2003. MMWR Morb Total Wkly Rep. 2003 52 (RR-17): 1-61. Chandler et al. , Antimicrob. Agents Chemother 1978; 14: 60-68. Guard et al. , Antimicrob Agents Chemother 1993; 37: 625-32. Chatterji S. Cement Concrete Res 1995; 25: 929-32. Clifton JC 2nd. Pediatr Clin North Am 2007; 54: 237-69. Codony et al. J Applied Microbiol 2003; 95: 288-93. Crane et al. , J Orthopaed Res 2009; 27: 1008-15. De Lalla, J Chemother. 2001; 13: 48-53. Depaola et al. . J Am Dent Assoc. 2002 Sep; 133 (9): 1199-206; quiz 1260. Dezelic et al. , Oral Health Prev Dent 2009; 7: 47-53. Domenico et al. , Canadian J. Microbiol. 31: 472-78 (1985). Domenico et al. , J Antimicrob Chemo 1991; 28: 801-810. Domenico et al. , Infection 20: 66-72 (1992). Domenico et al. , Infect. Immun. 62: 4495-99 (1994). Domenico et al. , J. et al. Antimicrol. Chemother. 38: 1031-40 (1996). Domenico et al. , Antimicrob Agents Chemother 1997; 41: 1697-703. Domenico et al. Infect Immun 67: 664-669 (1999). Domenico et al. 2000. Infect Med 17: 123-127. Domenico et al. , Antimicrob Agents Chemother 2001; 45: 1417-21. Domenico et al. , Research Advances in Antimicrob Agents Chemother 2003; 3: 79-85. Domenico et al. , J Antimicrob Chemo 1991; 28: 801-810; Domenico et al. , Infection 20: 66-72 (1992); Domenico et al. , Infect. Immun. 62: 4495-99 (1994); Domenico et al. , J. et al. Antimicrol. Chemother. 38: 1031-40 (1996); Domenico et al. , Antimicrob Agents Chemother 1997; 41: 1697-703; Domenico et al. ,
Infect Immun 67: 664-669 (1999); Domenico et al. 2000. Infect Med 17: 123-127; Domenico et al. , Antimicrob Agents Chemother 2001; 45: 1417-21; Domenico et al. , Research Advances in Antimicrob Agents Chemother 2003; 3: 79-85; Domenico et al. , J Antimicrob Chemo 1991; 28: 801-810. Domenico et al. , Peptides 2004. 25: 2047-53; Domenico et al. , 2005. Antibiotics for Clinicians 9: 291-297. Dufrene, J Bacteriol 2004; 186: 3283-85. Estefan et al. Gen Dent 2003; 51: 506-509. Fezel et al. , Proc. Natl. Acad. Sci. USA 106 (38): 16393-9. Epub 2009 Sep 14. Fulmer et al. , J Materials Sci: Materials Med 1992; 3: 299-305. Ganguli et al. , Smart Mater. Struct. 2009; 18: 104027. Geesey et al. (Eds) Biofouling and biocorrosion in industrial water systems. CRC Press, Boca Raton, FL, 1994. Gottenbos et al. , Biomaterials 2002; 23: 1417-23. Hamaguchi et al. Jap J Pharmacol 2000; 83: 273-76. Hu et al. , Study on injectable and degradable cement of calcium sulphate and calcium phosphate for bone repair. J Mater Sci Mater Med 2009 Oct 13. [Epub ahead of print], Huang et al. J Antimicrob Chemother 1999; 44: 601-605. Hwang et al. , Oral Surg Oral Med Oral Path Oral Radio Endo 2009; 107: e96-102. Idachaba et al. J Hazard Mater 2002; 90: 279-95. Idachaba et al. , Waste Manag Res 2001; 19: 284-91. Imazato,. Dent Materials 2003; 19: 449. Issa et al. , Oral Surg Oral Med Oral Path Oral Radio Endo 2004; 98: 553-65. Juhni et al. Proceedings Annual Meeting Adhesion Society 2005; 28: 179-181. Karchmer, Editorial Response: Clin Infect Dis 1998; 27: 714-16. Kavouras et al. , Inverteb Biol 2005; 122: 138-51. Kumar et al. , Nature Materials 2008; 7: 236-41. Leinfelder KF. JADA 2000; 131: 1186-87. Lovenoffer et al. , J Orthopaedic Trauma 2002; 16: 143-49. Mahony et al. , 1999 Antimicrob Agents Chemother 43: 582-88. Markarian J.M. Antimicrobials find new healthcare applications. Plastics, Additives and Compounding 2009; 11: 18-22. Masatoshi et al. , Development of antimicrobial plastics by Ag or Cu coatings sprayed via high velocity air fuel process. -Evaluation of the antimicrobial activity of Cu or Ag-sprayed plastics-Reports of the Shizuoka Industrial Research Institute 51. McDowell et al. J Am Dent Assoc 2004; 135: 799-805. Millsap et al. , Antonio Van Leeuwenhoek 2001; 79: 337-43. Omoike et al. , Biomacromolecules 2004; 5: 1219-30. Ouazzani et al. Congres 2008; 220: 290-94. Ozdamar et al. , Retina 1999; 19: 122-6. Piccilillo et al. J Mater Chem 2009; 19: 6167. Pitten et al. , Eur J Clin Pharmacol 1999; 55: 95-100. Reunala et al. Curr Opin Allergy Clin Immunol 2004; 4: 397-401. Rice et al. , Public Health Rep 2006; 121: 270-74. Romo et al. , Environ Progress 1999; 18: 107-12. Salo et al. , Infection 23: 371-77 (1995). Saha et al. , Cytokine modulation by bismuth-ethanediol in experimental sepsis. 10th Intl. Conf. Inflamm. Res. Assoc. , Hot Springs, VA. Sawada et al. , JPRAS 1990; 43: 78-82. Schultz, J Fluids Eng 2004; 126: 1039-47. Schultz MP, Biofouling 2007; 23: 331-41. Segreti et al. , Clin Infect Dis 1998; 27: 711-13. Sheffer, Am J Infect Cont 2005; 33: S20-5. Siboni et al. , FEMS Microbiol Lett 2007; 274: 24-29. Sidari et al. J Am. Water Works Assoc 2004; 96: 111-19. Soncini et al. JADA 2007; 138: 763-72. Stickelberg et al. , Prosthetic joint effects. In: Bisno et al. , Eds. Infections associated with indwelling medical devices. 2nd ed. Washington, DC: American Society for Microbiology, 1994: 259-90. Stoudley et al. , Clin Orthorelate Res 2005; 437: 31-40. Stout, ASHRAE J Oct 2007. Tazaki K.K. , Canadian Mineralogist 1992; 30: 431-34. Tiller et al. , Surface Coatings International Part B: Coatings Transactions 2005; 88: 1-82. Trachtenberg et al. J Dent Res 2009; 88: 276-79. Tsuneda et al. , FEMS Microbiol Lett 2003; 223: 287-92. Veloirah et al. , 2003, J Antimicrob Chemother 52: 915-19. Vu et al. Molecules 2009; 14: 2535-54. Widmer et al. J Infect Dis 1990; 162: 96-102. Widmer et al. , Antimicrob Agents Chemother 1991; 35: 741-46. Williams et al. , Compend Contin Educ Dent. 1996; 17: 691-94. Wu et al. , Am J .; Respir Cell Mol. Biol. 26: 731-38 (2002). Yan H, Li JO Phthalmolica 2008; 222: 245-48. Yeo et al. , Water Sci Techno 2007; 55: 35-42. Zardus et al. , Biol Bull 2008; 214: 91-98. Zarabi et al. J Oral Sci 2009; 51: 437-42. Zgonis et al. , J Foot Angle Surg 2004; 43: 97-103. Zhang et al. , 2005 Digestive Dis Sci 50: 1046-51; S. Patent No. 6, 582, 719; S. RE37,793; S. Patent No. 6, 248, 371; S. Patent No. 6,086,921; S. Patent No. 6,380,248; S. Patent No. 6, 582, 719; S. Patent No. 6,380,248; S. Patent No. 6,875,453.

  The various embodiments described above can be combined to provide further embodiments. All of the U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications and non-patent publications referenced herein and / or listed in the application data sheet are hereby incorporated by reference in their entirety. Incorporated into the book. The aspects of the embodiments can be modified as needed to provide further embodiments using various patent, application and publication concepts.

  These and other changes can be made to the embodiments in light of the above detailed description. In general, the terms used in the following claims should not be construed to limit the scope of the claims to the specific embodiments disclosed in the specification and the claims, and as such The claims should be construed to include all possible embodiments, along with the full scope of equivalents to which the claims are entitled.

Claims (35)

A method for protecting plants against bacterial, fungal or viral pathogens comprising:
(I) prevention of plant infection by bacterial, fungal or viral pathogens,
(Ii) inhibition of cell viability or cell proliferation of substantially all planktonic cells of bacterial, fungal or viral pathogens,
(Iii) inhibition of biofilm formation by bacterial, fungal or viral pathogens; and (iv) inhibition of biofilm viability or biofilm growth of substantially all biofilm type cells of bacterial, fungal or viral pathogens;
Contacting the plant with an effective amount of a bismuth-thiol (BT) composition under conditions and time sufficient for one or more of
The method, wherein the BT composition comprises a substantially monodispersed suspension of microparticles comprising a BT compound, wherein the microparticles have a volume average diameter of about 0.4 μm to about 10 μm.   The method of claim 1, wherein the bacterial pathogen comprises Erwinia amylobora cells.   The bacterial pathogens are Erwinia amylobora, Xanthomonas campestris pathogenic type dieffenbachiae, Pseudomonas syringae, Xylella fastidiosa; Xylophilus ampelinus; The method of claim 1, wherein the method is selected from the group consisting of sepedonics.   The method of claim 1, wherein the bacterial pathogen exhibits antibiotic resistance.   The method of claim 1, wherein the bacterial pathogen exhibits streptomycin resistance.   The method of claim 1, wherein the plant is an edible crop plant.   The method according to claim 6, wherein the food crop plant is a fruit tree.   8. The method of claim 7, wherein the fruit tree is selected from the group consisting of apples, pears, peaches, nectarines, plums and apricot trees.   The method according to claim 6, wherein the food crop plant is a genus Bancho.   The method according to claim 6, wherein the food crop plant is a plant selected from tuberous plants, legumes, and grain plants.   The method according to claim 10, wherein the tuberous root plant is selected from the group consisting of potato and sweet potato.   The method of claim 1, wherein the contacting step is performed one or more times.   13. The method of claim 12, wherein at least one of the contacting steps comprises one of spraying the plant, dipping the plant, coating the plant, and painting the plant.   13. The method of claim 12, wherein at least one step of contacting is performed at a plant bud, shoot or growth site.   13. A method according to claim 12, wherein at least one step of contacting is performed within 24, 48 or 72 hours of the first flowering in the plant.   The BT composition is BisBAL, BisEDT, Bis-dimercaprol, Bis-DTT, Bis-2-mercaptoethanol, Bis-DTE, Bis-Pyr, Bis-Ery, Bis-Tol, Bis-BDT, Bis-PDT Bis-Pyr / Bal, Bis-Pyr / BDT, Bis-Pyr / EDT, Bis-Pyr / PDT, Bis-Pyr / Tol, Bis-Pyr / Ery, Bismuth-1-mercapto-2-propanol and Bis-EDT The method of claim 1 comprising one or more BT compounds selected from the group consisting of / 2-hydroxy-1-propanethiol.   The method of claim 1, wherein the bacterial pathogen exhibits antibiotic resistance.   In connection with the step of contacting the plant with the BT composition, the method further comprises contacting the plant with a synergistic or potentiating antibiotic simultaneously or sequentially and in any order. 18. The method according to any one of items 17.   The synergistic or potentiating antibiotic is selected from the group consisting of aminoglycoside antibiotics, carbapenem antibiotics, cephalosporin antibiotics, fluoroquinolone antibiotics, penicillinase resistant penicillin antibiotics, and aminopenicillin antibiotics. The method of claim 18 comprising a selected antibiotic.   20. The synergistic or potentiating antibiotic is an aminoglycoside antibiotic selected from the group consisting of amikacin, arbekacin, gentamicin, kanamycin, neomycin, netilmycin, paromomycin, rhodostreptomycin, streptomycin, tobramycin and apramycin. The method described. A method for overcoming antibiotic resistance in a plant in which there is an antibiotic resistant bacterial phytopathogen, wherein:
(A) contacting the plant with an effective amount of BT composition under conditions and for a time sufficient for one or more of the following:
(I) prevention of plant infection by antibiotic-resistant bacterial pathogens,
(Ii) inhibition of cell viability or cell proliferation of substantially all planktonic cells of an antibiotic resistant bacterial pathogen,
(Iii) inhibition of biofilm formation by antibiotic-resistant bacterial pathogens; and (iv) inhibition of biofilm viability or biofilm growth of substantially all biofilm-type cells of antibiotic-resistant bacterial pathogens (in this case, The BT composition comprises a substantially monodispersed suspension of microparticles comprising a BT compound, the microparticles having a volume average diameter of about 0.5 μm to about 10 μm)
(B) A method comprising contacting a plant with a synergistic or potentiating antibiotic simultaneously or sequentially and in any order in connection with contacting the plant with the BT composition. A bismuth-thiol composition comprising a plurality of microparticles comprising a bismuth-thiol (BT) compound, wherein substantially all of the microparticles have a volume average diameter of about 0.4 μm to about 5 μm, and :
(A) a hydrophilic, polar or organic solubilizer comprising a bismuth salt containing bismuth at a concentration of at least 50 mM under conditions and time sufficient to obtain a solution substantially free of solid precipitate (Ii) admixing an acidic aqueous solution free of water with a sufficient amount of ethanol to obtain an admixture comprising about 25% by volume ethanol; and (b) an ethanolic solution comprising a thiol-containing compound (a) To obtain a reaction solution (wherein the thiol-containing compound is about 1: 3 relative to bismuth under conditions and time sufficient to form a precipitate containing microparticles containing the BT compound). Present in the reaction solution in a molar ratio of about 3: 1)
The method according to claim 1, which is formed by a process including   The method according to claim 22, wherein the bismuth salt is Bi (NO3) 3.   23. The method of claim 22, wherein the acidic aqueous solution comprises at least 5 wt%, 10 wt%, 15 wt%, 20 wt%, 22 wt% or 22.5 wt% bismuth.   Acidic aqueous solution at least 0.5 wt%, 1 wt%, 1.5 wt%, 2 wt%, 2.5 wt%, 3 wt%. 23. A process according to claim 22 comprising 3.5%, 4%, 4.5% or 5% by weight nitric acid.   Thiol-containing compound is 1,2-ethanedithiol, 2,3-dimercaptopropanol, pyrithione, dithioerythritol, 3,4-dimercaptotoluene, 2,3-butanedithiol, 1,3-propanedithiol, 2-hydroxy Propanethiol, 1-mercapto-2-propanol, dithioerythritol, α-lipoic acid, dithiothreitol, methanethiol (CH3SH [m-mercaptan]), ethanethiol (C2H5SH [e-mercaptan]), 1-propanethiol ( C3H7SH [n-P mercaptan]), 2-propanethiol (CH3CH (SH) CH3 [2C3 mercaptan]), butanethiol (C4H9SH [n-butyl mercaptan]), tert-butyl mercaptan (C (CH3) 3SH [t- Tilmercaptan]), pentanethiol (C5H11SH [pentylmercaptan]), coenzyme A, lipoamide, glutathione, cysteine, cystine, 2-mercaptoethanol, dithiothreitol, dithioerythritol, 2-mercaptoindole, transglutaminase, (11-mercapto Undecyl) hexa (ethylene glycol), (11-mercaptoundecyl) tetra (ethylene glycol), (11-mercaptoundecyl) tetra (ethylene glycol) functionalized gold nanoparticles, 1,1 ', 4', 1 ″ ′ -Terphenyl-4-thiol, 1,11-undecanedithiol, 1,16-hexadecanedithiol, technical grade 1,2-ethanedithiol, 1,3-propanedithiol, 1,4-benzenedi Methanethiol, 1,4-butanedithiol, 1,4-butanedithiol diacetate, 1,5-pentanedithiol, 1,6-hexanedithiol, 1,8-octanedithiol, 1,9-nonanedithiol, adamantanethiol 1-butanethiol, 1-decanethiol, 1-dodecanethiol, 1-heptanethiol, 1-heptanethiolpurum, 1-hexadecanethiol, 1-hexanethiol, 1-mercapto (triethylene glycol), 1 -Mercapto (triethylene glycol) methyl ether functionalized gold nanoparticles, 1-mercapto-2-propanol, 1-nonanethiol, 1-octadecanethiol, 1-octanethiol, 1-pentadecanethiol, 1-pentanethiol, 1 -Propanethio 1-tetradecanethiol, 1-tetradecanethiol plum, 1-undecanethiol, 11- (1H-pyrrol-1-yl) undecane-1-thiol, 11-amino-1-undecanethiol hydrochloride, 11-bromo-1 -Undecanthiol, 11-mercapto-1-undecanol, 11-mercaptoundecanoic acid, 11-mercaptoundecyl trifluoroacetate, 11-mercaptoundecyl phosphate, 12-mercaptododecanoic acid, 15-mercaptopentadecanoic acid, 16- Mercaptohexadecanoic acid, 1H, 1H, 2H, 2H-perfluorodecanethiol, 2,2 '-(ethylenedioxy) diethanethiol, 2,3-butanedithiol, 2-butanethiol, 2-ethylhexanethiol, 2- Methyl-1-propane All, 2-methyl-2-propanethiol, 2-phenylethanethiol, 3,3,4,4,5,5,6,6,6-nonafluoro-1-hexanethiolpurum, 3- (dimethoxymethylsilyl) -1-propanethiol, 3-chloro-1-propanethiol, 3-mercapto-1-propanol, 3-mercapto-2-butanol, 3-mercapto-N-nonylpropionamide, 3-mercaptopropionic acid, 3-mercapto Propyl-functionalized silica gel, 3-methyl-1-butanethiol, 4,4′-bis (mercaptomethyl) biphenyl, 4,4′-dimercaptostilbene, 4- (6-mercaptohexyloxy) benzyl alcohol, 4- Cyano-1-butanethiol, 4-mercapto-1-butanol, 6- (ferrocenyl) ) Hexanethiol, 6-mercapto-1-hexanol, 6-mercaptohexanoic acid, 8-mercapto-1-octanol, 8-mercaptooctanoic acid, 9-mercapto-1-nonanol, biphenyl-4,4′-dithiol, 3 -Butyl mercaptopropionate, copper 1-butanethiolate (I), cyclohexanethiol, cyclopentanethiol, decanethiol functionalized silver nanoparticles, dodecanethiol functionalized gold nanoparticles, dodecanethiol functionalized silver nanoparticles, Hexa (ethylene glycol) mono-11- (acetylthio) undecyl ether, mercaptosuccinic acid, methyl 3-mercaptopropionate, NanoTether BPA-HH, NanoThinks ™ 18, NanoThinks ™ 8, NanoT inks ™ ACID11, NanoThinks ™ ACID16, NanoThinks ™ ALCO11, NanoThinks ™ THIO8, octanethiol functionalized gold nanoparticles, PEG dithiol average Mn 8,000, PEG dithiol average molecular weight 1,500, PEG dithiol Average molecular weight 3,400, S- (11-bromoundecyl) thioacetate, S- (4-cyanobutyl) thioacetate, thiophenol, triethylene glycol mono-11-mercaptoundecyl ether, trimethylolpropane tris (3-mercaptopro Pionate), [11- (methylcarbonylthio) undecyl] tetra (ethylene glycol), m-carborane-9-thiol, p-terphenyl-4,4 ″ -di 23. The method of claim 22, comprising one or more agents selected from the group consisting of thiol, tert-dodecyl mercaptan, and tert-nonyl mercaptan. Said bacterial pathogens are:
(I) one or more gram-negative bacteria;
(Ii) one or more gram positive bacteria;
(Iii) one or more antibiotic-sensitive bacteria;
(Iv) one or more antibiotic-resistant bacteria;
(V) Staphylococcus aureus, MRSA (methicillin-resistant Staphylococcus aureus), Staphylococcus epidermidis, MRSE (methicillin-resistant Staphylococcus epidermidis), Mycobacterium tuberculosis, avian tuberculosis, Pseudomonas aeruginosa, drug-resistant Pseudomonas aeruginosa, Escherichia coli Enterotoxic Escherichia coli, Enterohemorrhagic Escherichia coli, Klebsiella pneumoniae, Clostridium difficile, Helicobacter pylori, Legionella pneumophila, Fecal streptococci, Methicillin sensitive fecal streptococci, Enterobacter cloacae, Salmonella typhimurium, Proteus bulgaris, Yersinia Enterocolitica, Vibrio cholerae, Shigella flexneri, Enterococcus vancomycin (VRE), Burkholderia cepacia group, wild gonorrhoeae, anthrax, plague, Pseudomonas aeruginosa, Streptococcus pneumoniae, Penicillin-resistant Streptococcus pneumoniae , E. coli, Burkholderia cepacia, bar Horuderia multi borane scan method according to any one of claims 1 to 21 comprising at least one of the bacterial pathogen is selected from the group consisting of M. smegmatis and Acinetobacter Baumani.   In connection with the step of contacting the surface with the BT composition, the plant can be treated simultaneously or sequentially, and in any order, with (i) a synergistic antibiotic and (ii) a cooperative antimicrobial efficacy enhancement. 22. A method according to any one of claims 1 to 21 comprising contacting with at least one of the sex antibiotics.   The synergistic antibiotic or the cooperative antibacterial efficacy enhancing antibiotic is an aminoglycoside antibiotic, a carbapenem antibiotic, a cephalosporin antibiotic, a fluoroquinolone antibiotic, a glycopeptide antibiotic, a lincosamide antibiotic 30. The method of claim 28, comprising an antibiotic selected from the group consisting of: a penicillinase resistant penicillin antibiotic, and an aminopenicillin antibiotic.   The synergistic antibiotic or the cooperative antibacterial efficacy potentiating antibiotic is an aminoglycoside antibiotic selected from the group consisting of amikacin, arbekacin, gentamicin, kanamycin, neomycin, netilmycin, paromomycin, rhodostreptomycin, streptomycin, tobramycin and apramycin 30. The method of claim 29, which is a substance. A method for overcoming antibiotic resistance in a plant in which there is an antibiotic resistant bacterial pathogen, wherein:
(I) prevention of plant infection by bacterial pathogens,
(Ii) inhibition of cell viability or cell proliferation of substantially all planktonic cells of the bacterial pathogen,
(Iii) inhibition of biofilm formation by bacterial pathogens; and (iv) inhibition of biofilm viability or biofilm growth of substantially all biofilm type cells of bacterial pathogens;
The plant is synergistically with an effective amount of (1) at least one bismuth-thiol (BT) composition and (2) at least one BT composition, under conditions and time sufficient for one or more of Including contacting with at least one antibiotic capable of enhancing or acting (wherein the BT composition is a bismuth-thiol composition comprising a plurality of microparticles comprising a bismuth-thiol (BT) compound) Wherein substantially all of the microparticles have a volume average diameter of about 0.4 μm to about 5 μm); and thereby overcoming antibiotic resistance of the epithelial tissue surface.   The said bacterial pathogen is resistant to an antibiotic selected from the group consisting of methicillin, vancomycin, nafcillin, gentamicin, ampicillin, chloramphenicol, doxycycline, tobramycin, clindamycin and gatifloxacin. The method described.   BT composition is BisBAL, BisEDT, Bis-dimercaprol, Bis-DTT, Bis-2-mercaptoethanol, Bis-DTE, Bis-Pyr, Bis-Ery, Bis-Tol, Bis-BDT, Bis-PDT, Bis-Pyr / Bal, Bis-Pyr / BDT, Bis-Pyr / EDT, Bis-Pyr / PDT, Bis-Pyr / Tol, Bis-Pyr / Ery, Bismuth-1-mercapto-2-propanol, and Bis-EDT 32. The method of claim 31, comprising one or more BT compounds selected from the group consisting of / 2-hydroxy-1-propanethiol.   Synergistic or potentiating antibiotics include clindamycin, gatifloxacin, aminoglycoside antibiotics, carbapenem antibiotics, cephalosporin antibiotics, fluoroquinolone antibiotics, penicillinase resistant penicillin antibiotics, and aminopenicillin 34. The method of claim 33, comprising an antibiotic selected from the group consisting of systemic antibiotics.   35. The synergistic or potentiating antibiotic is an aminoglycoside antibiotic selected from the group consisting of amikacin, arbekacin, gentamicin, kanamycin, neomycin, netilmycin, paromomycin, rhodostreptomycin, streptomycin, tobramycin and apramycin. The method described in 1.

Images