JP2020055839A - Bismuth-thiols as antiseptics for agricultural, industrial and other uses - Google Patents

Abstract

To provide bismuth-thiol compositions beneficially comprising substantially monodisperse microparticulate suspensions, and methods for their synthesis and use.SOLUTION: The present invention provides the use of bismuth-thiol (BT) compositions in producing drugs for treating open, chronic or acute wounds that were or are at risk for bacterial biofilm infections, where the BT compositions contain a monodisperse suspension of solid microparticles of bismuth-ethane dithiol (BisEDT) having a volumetric mean diameter of 0.4 μm to 5 μm.SELECTED DRAWING: Figure 2

Description

This application claims the benefit of PCT Application No. PCT / US2011 / 023549, filed February 3, 2010, and US Provisional Application No. 61 / 373,188, filed August 12, 2010. Claims, each of which is incorporated herein by reference in its entirety.

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

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

  Unfortunately, systemically or locally introduced antibiotics are often not effective for the treatment of many chronic bacterial infections and are generally not used unless acute bacterial infections are present. Current approaches include administration or application of antibiotics, but such remedies may promote the emergence of antibiotic-resistant strains and / or may be ineffective against bacterial biofilms . Therefore, it may be particularly important to use a disinfectant when drug-resistant bacteria (eg, methicillin-resistant Staphylococcus aureus or MRSA) are detected. Although there are many widely used disinfectants, established bacterial populations or subpopulations may not respond to these agents, or any other currently available treatment. In addition, multiple disinfectants can be toxic to host cells at the concentrations required to be effective against established bacterial infections, and such disinfectants are inappropriate. This problem may be due to natural surfaces, such as those of commercial and / or agriculturally important plants, such as many crop plants, and / or, for example, internal epithelial surfaces, such as respiratory tracts (eg, airways, nasopharynx and Attempts to purify infection from the laryngeal, tracheal, lung, bronchial, bronchiole, alveoli, etc. or gastrointestinal tracts (eg, oral cavity, esophagus, stomach, intestine, rectum, anus, etc.), or other epithelial surfaces In this case, it can be particularly serious.

  Of particular concern are infectious agents composed of bacterial biofilms, a relatively recently recognized bacterial tissue, in which planktonic unicellular (planktonic) bacteria differ markedly in their behavioral patterns and gene expression due to cell-cell adhesion. And an organized multicellular community (biofilm) that is sensitive to environmental drugs such as antibiotics. Biofilms may deploy biological defense mechanisms not found in planktonic bacteria, which can protect the biofilm community from antibiotics 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), Enterococci, Escherichia coli, Pseudomonas aeruginosa, and Streptococcus Genus and Acinetobacter baumannii. Some of these organisms exhibit the ability to survive on nutritionally ill clinical surfaces for several months. Staphylococcus aureus has been shown to be viable for 4 weeks on dried glass and 3-6 months on dried blood and cotton fibers (Domenico et al., 1999 Infect. Immun. 67: 664-). 669). Both E. coli and Pseudomonas aeruginosa have been shown to survive longer on dried blood and cotton fibers than S. aureus (ibid.).

  Microbial biofilms are associated with a substantial increase in resistance to both fungicides and antibiotics. Biofilm morphology results when bacteria and / or fungi attach to a surface. This attachment causes alterations in gene transcription, resulting in secretions that are significantly more active and difficult to penetrate the polysaccharide matrix, and protect the microorganism. Biofilms, in addition to having very substantial resistance to antibiotics, are very resistant to the mammalian immune system. Preventing biofilm formation is a very important clinical priority, as biofilms are very difficult to eradicate once settled. 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 severe wound infections.

  Intact and functioning skin and other epithelial tissues (e.g., commonly found in intact epithelial surfaces that form a barrier between an organism and its external environment, e.g., found in the skin, the respiratory and gastrointestinal tracts, The maintenance of those found in the lining, such as glandular tissue) is critical to the health and survival of humans and other animals.

Disinfectants based on bismuth thiol (BT) A number of natural products (e.g., antibiotics) and synthetic chemicals with antimicrobial, especially antibacterial, properties are known in the art; Antimicrobial effect, eg, the ability to kill microorganisms (“killing” effect such as bactericidal), the ability to stop or damage the growth of microorganisms (“resting” effect such as bacteriostatic), or microbial function, eg, It is characterized at least in part by colonization or infection, bacterial secretion of exopolysaccharides, and / or the ability to prevent conversion of planktonic populations to biofilm populations or expansion of biofilm formation. Antibiotics, bactericides, disinfectants, and the like (such as bismuth-thiol) that include factors that affect the selection and use of such compositions, such as, for example, bactericidal or bacteriostatic ability, effective concentrations, and the risk of toxicity to host tissues. Or containing a BT compound), for example, S. 6,582,719.

  Bismuth is a Group V metal and (like silver) is an antimicrobial element. Bismuth alone may not be therapeutically useful and may exhibit certain inappropriate properties, so that 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 a hyposalicylate. Combining certain thiol (-SH, sulfhydryl) -containing compounds, such as ethanedithiol and bismuth, to provide an exemplary bismuth thiol (BT) compound, provides an improved bismuth preparation compared to other bismuth preparations currently available. Improved research has determined that antimicrobial capacity is improved. There are many thiol compounds that can be used to make BT (eg, Domenico et al., 2001 Antimicrob. Agent. Chemotherap. 45 (5): 1417-14211, Domenico et al., 1997 Antimicrob. 41 (8): 1697-1703, and in US RE 37,793, US 6,248,371, 6,086,921, and 6,380,248. And see, for example, US Pat. No. 6,582,719), and some of these preparations can inhibit biofilm formation.

  BT compounds include MRSA (methicillin-resistant Staphylococcus aureus), MRSE (methicillin-resistant S. epidermidis), Mycobacterium tuberculosis, Mycobacterium avium, 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 Shigella flexneri have 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 microbes (gram-positive and gram-negative) that are resistant to broad spectrum antibiotics, prevent biofilm formation, and prevent septic shock However, it has also been demonstrated to treat sepsis and increase bacterial susceptibility to previously resistant antibiotics (eg, Domenico et al., 2001 Agent. Chemother. 45: 1417-1421; Domenico et al.). 17: 123-127; Antimicrob. Agents & Chemother. 3: 79-85, Domenico et al., 2003 Res. Adv .; Domenico et al., 2000 Infect. Microb.Agent.Chemother.41 (8): 1697-1703; Domenico et al., 1999 Infect.Immun.67: 664-669; Huang et al.1999 J Antimicrob.Chemother. 52, 915-919; Wu et al., 2002 Am J Respir Cell Mol Biol. 26: 731-738.), 2003 J Antimicrob.

Despite the availability of BT compounds for well over a decade, the effective selection of appropriate BT compounds for the indications of a particular infectious disease remains an elusive goal, with specific The behavior of BT cannot be predicted, the synergistic activity of a particular BT against a particular microorganism on a particular microorganism cannot be predicted, and the effect of BT in vitro can usually predict the BT effect in vivo. On the other hand, BT effects on planktonic (single cell) microbial populations cannot be predictive of BT effects on microbial communities, such as bacteria that have been assembled into biofilms. In addition, limitations such as solubility, tissue permeability, bioavailability, and biodistribution may hinder the ability to safely and effectively deliver clinical benefits for some BT compounds. There is. The embodiments of the invention disclosed herein address these needs and present other related benefits.
Plant and Agricultural Products Defense: Description of Related Technical Fields

  In the fields of agriculture and botany, the invention relates to formulations for reducing biofilms and diseases in plants and for example in seeds, plants, fruits and flowers, in soil and in cut flowers, trees, fruits, leaves, stems and other There is a recognized need for methods of using such formulations in the plant parts of the United States.

  In agriculture, billions of dollars of crops are lost each year due to biofilm formation. The problems of anthrax and biofilm-related diseases in plants are well understood, despite the number of approaches that have been attempted to address them failing to meet the needs. Plant diseases are involved in the transport and preservation of fruits, vegetables, cut flowers and trees, and other plant products, as the normal defense mechanisms used by intact living plants are no longer effective in harvested products It also affects the industries that do.

  Therefore, for agricultural purposes, reducing the amount of microbial growth on the surface of leaves, stems, fruits and flowers in situ, in transit or in a commercial context, 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 that the flow of water in cut flowers, plants and trees be able to maintain the swelling, integrity and quality of 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 consuming plant tissue and by exposing plant tissue to microorganisms.

  Biofilms typically form when bacteria bind to a surface in an aqueous environment, such as under water or in water droplets or other high humidity conditions, and after binding, the biofilm formation becomes sticky. It begins to secrete sexual substances, which in turn can bind to various materials, such as metals, plastics, medical implants and tissues. These biofilms can cause a number of problems including degradation of materials and plugging of pipes in box office and agricultural environments, and infection of surrounding tissues when occurring in a medical setting. The medical field is particularly vulnerable to problems caused by biofilm formation; implanted medical devices, catheters (urinary, venous, dialysis, heart) and slowly healing wounds are more easily affected by bacteria present in the biofilm. Infiltrated. In agriculture, biofilms can cause mastitis, Pierce's disease, ring rot of potatoes, downy mildew of various crops, and anthrax 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 surfaces and to each other via a complex matrix containing various extracellular polymeric substances (EPS), eg, exopolysaccharides, proteins and DNA. Plant-associated bacteria interact with host tissue surfaces during disease and symbiosis, and in a commensal relationship. Observations of bacteria associated with plants increasingly demonstrate a biofilm-type structure that changes from a small population of cells to a broad biofilm. Surface properties, nutrient and water availability of plant tissue, and the propensity of cloned bacteria strongly influence the resulting biofilm structure (Ramey et al., 2004 Curr Opinion Microbiol. 7: 602-9).

  The terrestrial environment is home to 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 form at the time of association or at a later stage and have significant potential to direct or regulate plant-microbial interactions. As many microorganisms actively modify the colonized plant environment, additional temporal and spatial complexity arises.

  Surface associated bacteria have significant effects on agriculture. In developed countries, losses caused by plant diseases have reached up to 25% of crop yields, the percentage of which is much higher than in developing countries. Epiphytic populations constitute a reservoir and future source of infection and can be found on host and non-host plants. Xylophilus ampelinus, a bacterial pathogen of grape vines, forms a thick biofilm in the vascular bundles of these plants (Grail & Manceau 2003). Pierre's disease (Xylella fastidiosa) is the causative agent of Pierce's disease in grape vines. Pierce fungi can form biofilms in the xylem tracts of many economically important crops. The pathogenic mechanism is largely due to the coagulation of Pierce's disease and occlusion of the xylem vessels by biofilm formation. Vessel obstruction is thought to play a major role in disease development, and xylem sap provides a natural medium that promotes the virulence of grapevine Pierce's disease and citrus variegated albinism (Zaini et al., 2009 FEMS Microbiol LETT). 295: 129-34).

  One of the most relevant plant pathogens, Pseudomonas syringae, causes brown spot on beans. It colonizes the leaf surface sporadically in a single subgroup (less than 10 cells), while the larger population (more than 1000 cells) is mainly due to trichomes with higher nutrient availability. Or develop near the veins. Large aggregates survive drought stress better than single cells. If it does not cause infection in host plant tissues, Pseudomonas syringes will remain alive as plant epiphytes (ie, colonies forming aerial parts of plants) (Monier et al., PNAS 2003; 100: 15977-82).

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

  Xanthomonas campestris pv. Campestris (Xcc) produces black roots in cruciferous plants and approaches the vascular bundle through the root wound site. Virulence involves degradable exoenzymes and exopolysaccharide xanthan gum required for virulence (Dow et al., PNAS 2003; 100: 10995-10000).

  Xanthomonas smithii subsp. Citri is the cause of citrus ulcer disease. The disease is found on most continents around the world outside Europe. The pathogen has been eliminated in many countries. Xanthomonas smithii forms ulcer lesions on the fruits, leaves and twigs of citrus plants. Wind and rain can spread bacteria up to 15 km away, breaking through stomata or wounds and transmitting to citrus trees (Sosnowski, et al., Plant Pathol 2009; 58: 621-35).

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

  Ralstonia solanacearum is a soil-borne pathogen that causes fatal mortality in many plants. Virulence is determined by 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 in potatoes. Showed an assemblage wrapped in a macrobacterial matrix associated with xylem vessels (Marques et al., Phytopathol 2003; 93: S57).

  Biofilm-producing Erwinia chrysanthemi causes soft rot due to the rapid decay of plant tissue. Pectin enzyme production is quorum sensing (QS) regulated, and therefore, the inability to form bacterial aggregates may render pectin degrading enzyme secretion impossible. A related plant pathogen, Erwinia amylovora, all infects about 75 different species of plants of the Rosaceae family. Hosts for this bacterium include apples, pears, blackberries, cotoneasters, wild apples, wildflower hawthorn (pyracantha), hawthorn, bokeh, rowan, pear, karin, raspberry, zaifribok and snowy willow. The cultivated varieties apple, pear and quince are the most severely affected species. A single burn epidemic in Michigan in 2000 lost more than 220,000 trees, resulting in a total loss of $ 42 million. Annual losses and management costs for burns in the United States are estimated to be in excess of $ 100 million (Norelli et al., Plant Dis 2003; 87: 26-32).

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

  If a part of the plant is attacked by a microbial plant pathogen, such as a biofilm former, the effect is usually to weaken or kill the plant. By infecting the leaves, the pathogen compromises the ability of the plant to produce its food (eg, by photosynthesis). Some plant pathogens block fluid transport tracts in the stalks that supply the leaves, and if such pathogens attack the roots, water and nutrient uptake is reduced or completely stopped. You. Blockage of plant vasculature is often accompanied by biofilm-producing bacteria that block the flow of water and nutrients, both in plants growing in soil as well as cut plants in vase water.

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

  Sometimes, a microbial "infection" is symbiotic, in which case both organisms benefit. A good example of this is the well-known nitrogen-fixing bacterium (rhizobia), which lives in the root nodules of legume (pea) plants, which provide nutrients and protection, while In turn, bacteria obtain nitrogen from the air and convert it into a form that can be used by the host. As another example, mycorrhiza is one large eye of fungi that has a symbiotic relationship with plant roots. In view of such mutually beneficial symbiosis, protection or protection of plants against harmful microbial pathogens desirably uses antimicrobial agents that do not disrupt these symbiotic relationships whenever possible.

  Saprophytic fungi are essential for breaking down dead biological material and producing humus, which is required for good soil structure. They do not have any chlorophyll and therefore cannot use light to capture energy (eg, by photosynthesis); instead, they use plant and animal matter (live or dead) To obtain its energy. Although they can also survive in symbiosis with the thin root mycorrhiza of certain plant species, for example conifers, they cannot survive without them to take up nutrients necessary for life support. The widespread use of chemicals to control harmful plant pathogens compromises 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 weaken or kill them. Viruses, another class of microbial plant pathogens, live in the cells of plant tissues and are therefore often inaccessible to topically applied chemicals, leaving diseased plants inevitable. In general, no antibiotics have been specifically developed for the treatment of plants (although some antibiotics developed for other purposes find use in plants) and a large number of economically significant Of plant species remain susceptible to pathogenic bacterial attack. For example, the infestation of many plant species in the family Rosaceae has proven incurable. In contrast, a number of harmful fungi can be killed with topically applied chemicals without harming the plant host, because of the different fungal growth behavior, i.e., a number of undesirable pathogenic This is because fungi use root-like structures to extract nutrients and grow on plant surfaces but not in plant tissues.

  Because it is often difficult or impossible to kill many plant pathogens, many strategies for protecting plants against harmful microbial pathogens favor 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, if such agents are used prophylactically, rather than in response to established infections, significantly lower amounts of insecticides or microbicides may be effective.

For example, if the plant is not growing under optimal or near-optimal conditions because the soil is poor in quality (eg, lack of nutrients) by itself or in combination with drought or heavy rain or flood, They are also more susceptible to disease. For example, extreme humid conditions may promote pathogenic fungal and / or bacterial growth. For example, quorum sensing in Pseudomonas syringes is indicated by leaf surface water availability (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 juice sucking insects are the major vectors of the virus. Fungal disease spores are carried in the air and in raindrops and droplets.
Biofilm on seeds and shoots

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

  Vascular pathogens live in the xylem or phloem of the plant host and generally depend on insect vectors or wounds involved in transmission. Cut flowers or cut trees are similar types of wounds that are particularly prone to vascular infection. Biofilm bacteria enter and clog the vascular bundles on the cut surface, obstructing the flow of water, minerals and nutrients. Cut flower preservatives diluted in vase water often contain salicylates 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 biofilms.

  Antibacterial agent in agriculture. Eradication of plant pathogen invasion is very important for the protection of the plant industry, managed gardens and the natural environment worldwide. The consequences of endemic pathogens are severe and, in some cases, can affect a country's economy. Current strategies for the eradication of pathogens rely on techniques for treatment, removal and disposal of diseased host plants. There are numerous examples where these techniques have been successful, but often not. 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 diseased hosts around the world, especially in Australia, various techniques such as burning, filling, composting, soil and biofumigation, solarization, steam sterilization and biological vehicles Control has 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 for plants are oxytetracycline and streptomycin. In the United States, antibiotics applied to plants account for less than 0.5% of total antibiotic use. The resistance of phytopathogens to oxytetracycline is rare, but the emergence of streptomycin-resistant strains of Erwinia amilobora, Pseudomonas species and Xanthomonas campestris has impeded control of several important diseases. Therefore, the role of antibiotic use in plants in the antibiotic resistance crisis in human medicine is the subject of controversy (McManus et al., Annu Rev Phytopathol 2002; 40: 443-65).

  The emergence of streptomycin resistant (SmR) plant pathogens complicates the control of bacterial diseases in plants. For example, in the United States, streptomycin is available from Xanthomonas campestris pv. It has been licensed for tomato and pepper for control of vesica tria, but is rarely used for this purpose because the resistant strains are now widespread. Resistance in the burn pathogen Erwinia Amilobola has widespread economic and political implications. Other phytopathogenic bacteria for which SmR has been reported include Pectobacterium carotobola, Pseudomonas chicory, Pseudomonas lacrimans, Pseudomonas syringe pv. Paprance, Pseudomonas syringe pv. Syringe and Xanthomonas dieffenbachie (McManus et al., Annu Rev Phytopathol 2002; 40: 443-65). The emergence of SmR Erwinia amilobora has increased the burn epidemic in the western United States and Michigan.

  Streptomycin and oxytetracycline have been assigned to the lowest toxicity category by the United States Environmental Protection Agency (EPA), and no carcinogenic and mutagenic activity has been observed for either antibiotic.

  Alternatives to antibiotics are available and practical, at least to some extent. In fact, bacterial disease management in most crop systems is based on the integration of host genetic resistance, sanitation (avoidance or elimination of inoculation), and cultivation practices that create an unfavorable environment for disease development. Biocontrol of plants using various bacterial and fungal species is of increasing interest. Rhizobia are considered efficient microbial competitors in the root zone. Representatives of a number of different bacterial genera have been introduced into soil, on seeds, roots, tubers or other plant material to improve crop growth. Examples of these bacterial genera include Acinetobacter, Agrobacterium, Arthrobacter, Azospirillum, Bacillus, Bradyrhizobium, Frankia, Pseudomonas, Rhizobium, Serratia, Thiobacillus and the like. For example, certain Bacillus can induce systemic tolerance in many plants (Choudhary & Johr. Microbiol Res 2009; 164: 493-513).

  The 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.

  Numerous synthetic and natural therapies exist for various plant diseases. Natural remedies include apple vinegar for lesions, mildew and mildew; spraying baking soda for anthrax, early tomato blight, leaf blight, powdery mildew, and as a common fungicide; Neem oil; sulfur; garlic; hydrogen peroxide; compost tea and the like. Numerous synthetic chemicals are used to prevent or treat plant diseases, resulting in water-soluble or water-insoluble formulations. Fungicides include phenoxyarsine or phenalsadine, maleimide, isoindoledicarboximide, aryl alkanol halides, 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 isothiazolecarboxamides can be used for control of plant pests (eg, US Pat. No. 6,525,056; WO 2001/064644).

  Understanding the toxicity problems of fungicides in powder or crystalline form, U.S. Patent No. 29,409 discloses dissolving a fungicide in a liquid solvent, adding it to a formulation mixture, and then adding the end-use resin composition. It teaches that objects can be processed. The dispersion can be used safely where the end use resin composition is prepared, but inadvertent use or disposal of the liquid can still result in environmental and health hazards. Alternatively, the bactericide can be administered in a water soluble thermoplastic. A disinfectant can be added to the rigid thermoplastic composition to confer biocidal activity to it to inhibit microbial growth on its surface (US Pat. No. 5,229,124). It is a solid melt compounding solution consisting essentially of a germicide dissolved in a carrier resin that is a copolymer of vinyl alcohol and (alkyleneoxy) acrylate. The fungicide can be a highly toxic chemical, but its low concentration in the end use product as well as its retention by the resin composition means that the antimicrobial in the end use product does not harm humans or animals. Guarantee.

  Isothiazolinones, such as N-alkylbenzenesulfonylcarbamoyl-5-chloroisothiazole derivatives, are often used as fungicides in agriculture (eg, US Pat. No. 5,045,555). This disinfectant is used, for example, in the paper and textile industries, to produce coatings and adhesives, in painting, metalworking, in the resin, timber, building, agriculture, forestry, fisheries, food and petroleum industries. , As well as in medicine. It exhibits strong antibacterial action and can be added to industrial water, circulating water, raw materials or products in appropriate amounts. Further, it can be used to disinfect and sterilize equipment, industrial facilities, livestock pens or equipment, as well as 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 also generally 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 the soft rot and burns induced by Erwinia chrysantemi and Erwinia amilobora, respectively (Expert. Annu Rev Phytopathol 1999; 37: 307). -34). Because of its unique position in biological systems, iron controls the activity of plant pathogens. The production of siderophores by pathogens represents not only 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 development is another major problem. Antimicrobial 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, at least in part chemically and biologically. Exemplary properties include the ability to kill microorganisms (bactericidal action), the ability to stop or impair microbial growth (bacteriostatic action), or the bacterial nature of metabolites that colonize or infect a site. The ability to secrete (some of which stinks) and / or interfere with microbial functions such as conversion from planktonic populations to biofilm populations or biofilm expansion (anti-biofilm effects). Antibiotics, disinfectants, bactericides and the like (including bismuth-thiol or BT compounds) are factors that influence the selection and use of such compositions, such as bactericides, bacteriostats or anti-biofilm efficacy, It is discussed in U.S. Patent No. 6,582,719, including effective concentrations and the danger of toxicity to host tissues.

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

  Repression during the rot stage is important for the management of burns. Erwinia amilobora needs to grow on the stigmatic surface during the epigenesis in order to cause flower infection. Rain is necessary for infection as it dilutes the sugar on the flower sac to an osmotic potential that is not inhibitory for Erwinia amilobora. Rain is also an important agent for the redistribution of bacteria from stigmas to flower canals. These observations suggest that the optimal time to use antibiotic spray is during this episode, as well as after heavy rain (Johnson & Stockwell. Annu Rev Phytopathol 1998; 36: 227-48). .

  Other bacterial epiphytes also colonize stigmas, where they can interact with and repress epiphytic growth of pathogens. A commercial bacterial antagonist from Erwinia amilobora (BrightBan, Pseudomonas fluorescens A506) can be included in the antibiotic spray program. Incorporation of bacterial antagonists by chemical methods suppresses the population of pathogens and co-localizes the non-pathogenic competitors in the ecological niches provided by the stigmas (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 coordination complex of zinc. This colorless solid is used as an antifungal and antibacterial agent. Due to its low water solubility (8 ppm at neutral pH), zinc pyrithione is suitable for use in outdoor applications, cement and other products that provide protection against mildew and algae. It is an effective algicide. However, it is chemically incompatible with the amount when relying on metal carboxylate hardeners. When used in latex paints containing water containing high amounts of iron, sequestering agents that selectively bind iron ions are required.

  Of particular concern in agriculture are infections that consist of bacterial biofilms, a rather recently recognized organization of bacteria, whereby free unicellularized ("planktonic") bacteria are transformed by cellular indirects. The dressing assembles into an organized multicellular community (biofilm) with markedly different behavioral patterns, gene expression, and susceptibility to environmental agents, including antibiotics. Biofilms develop a biological defense mechanism not found in planktonic bacteria, which may 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 a surface. This attachment triggers a transcriptional alteration of the gene, resulting in the secretion of a polysaccharide matrix that is remarkably elastic and difficult to penetrate, protecting the microorganism. Biofilms, besides their very substantial resistance to antibiotics, are very resistant to plant immune defense mechanisms. Biofilms, once established, are very difficult to eradicate, 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, clarification, and / or wound healing, and are likely to be largely linked to 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 occurring 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, the bismuth-thiol (BT) compounds may be used in a wide variety of agricultural While it can be used as an antiseptic for use in industrial, manufacturing and other settings, and in the treatment or prevention of infectious diseases and related conditions, and in personal health care for personal health care, It also reduces the cost of treating such infections, including savings made 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, plants or plant tissues (e.g., roots, bulbs, stems, leaves, branches, vines, creeping branches, buds, flowers or parts thereof, including plants that can form or otherwise promote biofilms), sprout , Fruits, seeds, pods, etc.) and formulations for treating animal tissues and / or natural and artificial surfaces are contemplated, and according to non-limiting theory, appropriately selected BT compounds based on the present disclosure. The combination of antibiotic (s) with the antibiotic (s) may have previously unanticipated synergy 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 are bismuth-thiol compositions that advantageously comprise a substantially monodisperse particulate suspension, and methods of their synthesis and use. It is.

  Thus, according to certain embodiments of the invention described herein, a method for protecting a plant against a bacterial, fungal or viral pathogen, comprising: (i) a bacterial, fungal or viral pathogen Preventing plant infection, (ii) inhibiting cell viability or cell growth of substantially all planktonic cells of a bacterial, fungal or viral pathogen, (iii) inhibiting biofilm formation by a bacterial, fungal or viral pathogen And (iv) inhibiting the biofilm viability or biofilm growth of substantially all biofilm-type cells of a bacterial, fungal or viral pathogen, under sufficient conditions and time for one or more of the following: Or a part thereof (eg, root, bulb, stem, leaf, branch, vine, creeping branch, bud, flower or a part thereof, sprout, fruit, seed, Etc.) with an effective amount of a bismuth-thiol (BT) composition, wherein the BT composition comprises a substantially monodisperse suspension of fine particles comprising the BT compound. Provided is a method, 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 amilobora cells. In another embodiment, the bacterial pathogen is Erwinia amilobolla, Xanthomonas campestris pathogen dieffenbachiae, Pseudomonas syringae, Xylella fastiidosa; Xylophilus amperinus; It is selected from Bacter Michiganensis subspecies sepedonics. In certain embodiments, the bacterial pathogen exhibits antibiotic resistance. In certain embodiments, the bacterial pathogen exhibits streptomycin resistance. In some embodiments, the plant is a food crop plant, and in some further embodiments, the food crop plant is a fruit tree. In certain further embodiments, the fruit tree is selected from an apple, pear, peach, nectarine, plum, apricot tree. In one embodiment, the food crop plant is a banana tree of Musa. In another embodiment, the food crop plant is a plant selected from tuberous root plants, legumes, and grain plants. In a further embodiment, the tuberous plant is selected from potato (potato) and sweet potato (sweet potato). In certain embodiments of the above method, the contacting step is performed one or more times. In some further embodiments, the at least one step of contacting comprises one of spraying the plant, dipping the plant, coating the plant, and painting the plant. In certain other further embodiments, at least one step of contacting is performed at a flower bud, shoot, or growth site of the plant. 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 method, 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- Includes 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 a further embodiment of the above method, the method comprises synergistic or potentiating the plant, simultaneously or sequentially, and in any order, in connection with the step of contacting the plant with the BT composition. Contacting the 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, netilmycin, paromomycin, rhodostreptomycin, streptomycin, tobramycin, and apramycin.

  According to certain other embodiments, a method for overcoming antibiotic resistance in a plant in which an antibiotic-resistant bacterial plant pathogen is present, comprising: (a) one of the following: Contacting the plant with an effective amount of the BT composition under conditions and for a time sufficient for one or more of: (i) prevention of infection of the plant by the antibiotic-resistant bacterial pathogen, (ii) substance of the antibiotic-resistant bacterial pathogen. Inhibiting the cell viability or proliferation of all planktonic cells, (iii) inhibiting biofilm formation by antibiotic-resistant bacterial pathogens, and (iv) substantially all biofilm types of antibiotic-resistant bacterial pathogens Inhibition of biofilm viability or biofilm growth of cells (where the BT composition comprises a substantially monodisperse 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 conjunction with contacting the plant with the BT composition, simultaneously or sequentially, and in any order Contacting the plant with a synergistic or potentiating antibiotic.

  In certain embodiments of the above method, 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 from about 0.4 μm to about 5 μm. And (a) a bismuth salt comprising a bismuth salt having a concentration of at least 50 mM, under conditions and for a time sufficient to obtain a solution substantially free of solid precipitates, Mixing an acidic aqueous solution free of polar or organic solubilizers with (ii) a sufficient amount of ethanol to obtain a blend containing about 25% by volume of ethanol; and (b) mixing the thiol-containing compound with The ethanolic solution containing the thiol-containing compound is added to the mixture of (a) to obtain a reaction solution (in this case, the thiol-containing compound is formed under conditions sufficient to form a precipitate containing fine particles containing the BT compound). (In a molar ratio of about 1: 3 to about 3: 1 with respect to bismuth, in the reaction solution).

  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 of bismuth. In certain embodiments, the acidic aqueous solution is at least 0.5%, 1%, 1.5%, 2%, 2.5%, 3% by weight. It contains 3.5%, 4%, 4.5% or 5% by weight of 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-butanedithiol diacetate, 1,5-pentanedithiol, 1,6-hexanedithiol, 1,8-octanedithiol, 1,9 -Nonanedithiol, adamantanethiol, 1-butanethiol, 1-decanethiol, 1-dodecanethiol, 1-heptanethiol, 1-heptanethiol purum, 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-mercaptoundecyltrifluoroacetate, 11-mercaptoundecylphosphoric acid, 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-hexanethiol plum, 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, butyl 3-mercaptopropionate, copper (I) 1-butanethiolate, cyclohexanethiol, cyclopentanethiol, decanethiol functionalized silver nanoparticles, dodecanethiol functionalized gold nanoparticles, dodecanethiol functionalization Silver nanoparticles, hexa (ethylene glycol) mono-11- (acetylthio) undecyl ether, mercaptosuccinic acid, methyl 3-mercaptopropionate, NanoTeether BPA-HH, NanoThinks (TM) 18, NanoThinks (trademark) ) 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-terphenyl And at least one drug selected from -4,4 "-dithiol, tert-dodecylmercaptan, and tert-nonylmercaptan.

  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 More than one antibiotic resistant bacteria; (v) Staphylococcus aureus, MRSA (Methicillin-resistant Staphylococcus aureus), Staphylococcus epidermidis, MRSE (Methicillin-resistant Staphylococcus epidermidis), Mycobacterium tuberculosis, Mycobacterium avium, 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 Bacteria, Proteus vulgaris, Yersinia enterocolitica, cholera, Shigella flexneri, van Mycin-resistant enterococcus (VRE), Burkholderia cepacia, group of tularemia, anthrax, pestis, Pseudomonas aeruginosa, Streptococcus pneumoniae, penicillin-resistant Streptococcus pneumoniae, Escherichia coli, Burkholderia cepacia, Burkholderia -Comprising at least one of the following: a bacterial pathogen selected from Multiborans, Syphilis and Acinetobacter baumannii.

  In certain embodiments, the method comprises, simultaneously or sequentially, and in any order, relating to the step of contacting the surface with the BT composition, comprising: (i) synergistic antibiotic and (Ii) contacting with at least one of the cooperative antimicrobial potency enhancing antibiotics. In certain further embodiments, the synergistic or cooperative antimicrobial potency enhancing antibiotic is an aminoglycoside antibiotic, a carbapenem antibiotic, a cephalosporin antibiotic, a fluoroquinolone antibiotic, a glycopeptide antibiotic. Lincosamide antibiotics, penicillinase-resistant penicillin antibiotics, and aminopenicillin antibiotics. In certain further embodiments, the synergistic or cooperative antimicrobial potency enhancing antibiotic is an aminoglycoside selected from amikacin, arbekacin, gentamicin, kanamycin, neomycin, netilmycin, paromomycin, rhodostreptomycin, streptomycin, tobramycin and apramycin. It is a system antibiotic.

  In certain other embodiments, a method for overcoming antibiotic resistance in a plant having an antibiotic resistant bacterial pathogen therein or thereon, comprising: (i) preventing infection of the plant by the bacterial pathogen; (Ii) inhibition of cell viability or cell growth of substantially all planktonic cells of the bacterial pathogen, (iii) inhibition of biofilm formation by the bacterial pathogen, and (iv) substantially all biofilms of the bacterial pathogen Inhibiting Biofilm Viability or Biofilm Growth of Type Cells: Under conditions and for a time sufficient for one or more of the following, an effective amount of (1) at least one bismuth-thiol (BT) composition And (2) contacting with at least one antibiotic that can synergistically enhance or act with at least one BT composition. (In this case, the BT composition is a bismuth-thiol composition including a plurality of fine particles including a bismuth-thiol (BT) compound, and substantially all of the fine particles have a particle size of about 0.4 μm to about 5 μm. And thereby overcoming antibiotic resistance on the epithelial tissue surface. In certain further embodiments, the bacterial pathogen exhibits resistance 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. In certain embodiments, the synergistic or potentiating antibiotic is clindamycin, gatifloxacin, an aminoglycoside antibiotic, a carbapenem antibiotic, a cephalosporin antibiotic, a fluoroquinolone antibiotic, a penicillinase resistant penicillin antibiotic. Substances, and aminopenicillins. In a further embodiment, the synergistic or potentiating antibiotic is an aminoglycoside antibiotic selected from amikacin, arbekacin, gentamicin, kanamycin, neomycin, netilmycin, paromomycin, rhodostreptomycin, streptomycin, tobramycin and apramycin. .

  According to certain other embodiments, there is provided a bismuth-thiol composition comprising a plurality of microparticles comprising a bismuth-thiol (BT) compound, 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 bismuth or a bismuth salt and a thiol-containing compound. In another embodiment, there is provided 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 from about 0.4 μm to about 5 μm. (I) comprising a bismuth salt comprising bismuth at a concentration of at least 50 mM, under conditions and for a time sufficient to obtain a solution substantially free of solid precipitates; The acidic aqueous solution without the solubilizing agent is sufficiently mixed with (ii) a mixture comprising at least about 5%, 10%, 15%, 20%, 25% or 30% by volume of ethanol. And (b) a molar ratio of about 1: 3 to about 3: 1 with respect to bismuth under conditions and for a time sufficient to form a precipitate containing fine particles 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%, 10%, 15%, 20%, 22%, or 22.5% by weight of bismuth. In certain embodiments, the acidic aqueous solution is at least 0.5%, 1%, 1.5%, 2%, 2.5%, 3% by weight. It contains 3.5%, 4%, 4.5% or 5% by weight of 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- Contains one or more agents selected from propanedithiol, 2-hydroxypropanethiol, 1-mercapto-2-propanol, α-lipoic acid, and dithiothreitol.

  In another embodiment, there is provided a method of preparing a bismuth-thiol composition comprising a plurality of microparticles comprising a bismuth-thiol (BT) compound, wherein substantially all of the microparticles comprise from about 0.4 μm to about 5 μm. Having a volume average diameter and wherein the method comprises the steps of: (i) forming a bismuth salt comprising a bismuth salt having a concentration of at least 50 mM under conditions and for a time sufficient to obtain a solution substantially free of solid precipitates; (Ii) at least about 5%, 10%, 15%, 20%, 25% or 30% by volume of ethanol containing an aqueous acidic solution containing no hydrophilic, polar or organic solubilizers. Admixing with a sufficient amount of ethanol to obtain a mixture containing the BT compound; and (b) adding bismuth to the bismuth under conditions and for a time sufficient to form a precipitate containing fine particles containing the BT compound. Adding an ethanolic solution containing the thiol-containing compound present in the reaction solution in a molar ratio of about 1: 3 to about 3: 1 to the mixture of (a) to obtain a reaction solution. In certain embodiments, the method further comprises collecting 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%, 10%, 15%, 20%, 22%, or 22.5% by weight of bismuth. In certain embodiments, the acidic aqueous solution is at least 0.5%, 1%, 1.5%, 2%, 2.5%, 3% by weight. It contains 3.5%, 4%, 4.5% or 5% by weight of 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 Butylmercaptan]), pentanethiol (C5H11SH [pentylmercaptan]), 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, , 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-heptanethiol purum, 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-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-mercaptoundecyltrifluoroacetate, 11-mercaptoundecylphosphoric acid, 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-hexanethiolplum, 3- (dimethoxymethylsilyl) -1-propanethiol, -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 It contains one or more drugs selected from the group consisting of decyl mercaptan and tert-nonyl mercaptan.

  In another embodiment, (i) preventing infection of the surface by a bacterial, fungal or viral pathogen; (ii) inhibiting cell viability or cell development of substantially all planktonic cells of the bacterial, fungal or viral pathogen; (Iii) inhibition of biofilm formation by bacterial, fungal or viral pathogens; and (iv) inhibition of biofilm viability or biofilm development of substantially all biofilm-type cells of bacterial, fungal or viral pathogens. Contacting the epithelial tissue surface with an effective amount of a BT composition under conditions and for a time sufficient for one or more of the following: 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 originals, natural or artificial surfaces, such as biological tissue surfaces, such as plant surfaces (eg, roots, bulbs, stems, leaves, branches, vines, creeping branches, buds, flowers or parts thereof, shoots , Fruits, seeds, pods, etc.) or a surface of epithelial tissue. 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 (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 Fekaris, Methicil Susceptible Enterococcus faecalis, Enterobacter cloacae, Salmonella typhimurium, Proteus vulgaris, Yersinia enterocolitica, Vibrio cholere, Shigella flexneri, vancomycin-resistant Enterococcus (VRE), Burkholderia cepacia, Francisella tularensis, Bacillus anthracis, Yersinia pestis, Pseudomonas aeruginosa, Vancomycin-resistant Enterococcus sp., Streptococcus pneumoniae, Penicillin-resistant Streptococcus pneumoniae, Escherichia coli, Burkholderia cepacia, Burkholderia multibolans・ A small number of bacterial pathogens selected from smegmatis and Acinetobacter baumanii 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 an oral / oral surface, an artificial device, ceramic, plastic, polymer, rubber, metal product, painted surface, marine structure, such as a hull, rudder, propeller, anchor, hold, Including ballast tanks, docks, dry docks, piers, piles, seawalls, or other natural or artificial surfaces.

  In certain embodiments, the surface comprises an epithelial tissue surface including a tissue selected from the epidermis, dermis, respiratory tract, gastrointestinal tract and gland lining.

  In certain embodiments, the step of contacting is performed one or more times. In certain embodiments, the at least one contacting step comprises one of spraying, irrigating, dipping, and painting a natural or artificial surface. In certain embodiments, the at least one contacting step comprises one of inhalation, ingestion, and oral irrigation. In certain embodiments, at least one contacting step comprises topical, intraperitoneal, oral, parenteral, intravenous, intraarterial, transdermal, sublingual, subcutaneous, intramuscular, buccal, intranasal, inhalation, ocular Intravenous, intra-aural, intra-ventricular, intra-fat, intra-articular, and intrathecal routes. In certain embodiments, 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- Contains one or more BT compounds selected from the group consisting of propanol and Bis-EDT / 2-hydroxy-1-propanethiol.

  In certain embodiments, the bacterial pathogen exhibits antibiotic resistance. In certain other embodiments, the method comprises synthesizing the natural surface with a synergizing antibiotic and / or in any order simultaneously or sequentially with the step of contacting the natural or artificial surface with the BT composition. Further comprising contacting with an enhancing antibiotic. In certain embodiments, the synergistic and / or potentiating antibiotic is an aminoglycoside antibiotic, a carbapenem antibiotic, a cephalosporin antibiotic, a fluoroquinolone antibiotic, a glycopeptide antibiotic, a lincosamide antibiotic. It includes antibiotics selected from antibiotics, penicillinase-resistant penicillins, and aminopenicillins. 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) preventing 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 the cells, (iii) inhibition of biofilm formation by bacterial pathogens, and (iv) inhibition of biofilm viability or biofilm development of substantially all biofilm-type cells of bacterial pathogens Under conditions and for a time sufficient for one or more of the following, to provide an effective amount of (1) at least one bismuth-thiol (BT) composition and (2) simultaneously with at least one antibiotic that is capable of being enhanced by and / or synergistic with at least one BT composition. Comprises continuously contacting in any order, 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 natural or artificial surfaces where antibiotic resistant bacterial pathogens are present (e.g., bacteria of the same species For bacterial pathogens that are resistant to at least one antimicrobial effect of at least one antibiotic known to have an antimicrobial effect against such pathogens). . 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 (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 Fekaris, Methicil Susceptible Enterococcus faecalis, Enterobacter cloacae, Salmonella typhimurium, Proteus vulgaris, Yersinia enterocolitica, Vibrio cholere, Shigella flexneri, vancomycin-resistant Enterococcus (VRE), Burkholderia cepacia, Francisella tularensis, Bacillus anthracis, Yersinia pestis, Pseudomonas aeruginosa, Vancomycin-resistant Enterococcus sp., Streptococcus pneumoniae, Penicillin-resistant Streptococcus pneumoniae, Escherichia coli, Burkholderia cepacia, Burkholderia multibolans・ A small number of bacterial pathogens selected from smegmatis and Acinetobacter baumanii 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 oral / oral surface, an artificial device, a ceramic, plastic, polymer, rubber, metal product, painted surface, marine structure, such as a hull, tire, propeller, anchor, hold, Including ballast tanks, docks, dry docks, piers, piles, seawalls, 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 lining of glands. In certain embodiments, the step of contacting is performed one or more times. In certain embodiments, the at least one contacting step comprises one of spraying, irrigating, dipping, and painting the surface. In certain other embodiments, the at least one contacting step comprises one of inhalation, ingestion, and oral irrigation. In certain embodiments, at least one contacting step comprises topical, intraperitoneal, oral, parenteral, intravenous, intraarterial, transdermal, sublingual, subcutaneous, intramuscular, buccal, intranasal, inhalation, ocular Intravenous, intra-aural, intra-ventricular, intra-fat, intra-articular, and intrathecal routes. In certain embodiments, 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- It contains one or more BT compounds selected from propanol and Bis-EDT / 2-hydroxy-1-propanethiol. In certain embodiments, the synergistic and / or potentiating antibiotic is clindamycin, gatifloxacin, aminoglycoside antibiotic, carbapenem antibiotic, cephalosporin antibiotic, fluoroquinolone antibiotic, It includes antibiotics 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 enhanced by and / or synergistic with the BT compound; and (C) An antiseptic 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-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, wherein substantially all of the microparticles have a volume average diameter from 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) the bacterial infection on or within the surface is (i) gram-positive, (ii) gram-negative, and (iii) both (i) and (ii) And b) administering to the surface a formulation comprising one or more bismuth thiol (BT) compositions, the method comprising supporting or containing a bacterial biofilm. Alternatively, there is provided a method of treating an artificial surface, wherein (i) if the bacterial infection comprises a Gram-positive bacterium, the formulation comprises at least one BT compound and at least one antibiotic, rifamycin. 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) the 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 the bacterial biofilm, (ii) reducing the bacterial biofilm, and (iii) inhibiting the growth of the bacterial biofilm. At least one of them. In certain embodiments, the BT composition comprises a plurality of microparticles comprising a bismuth-thiol (BT) compound, wherein substantially all of the microparticles have a volume average diameter from about 0.4 μm to about 5 μm.

  These and other aspects of the embodiments of the invention described herein will become apparent with reference to the following detailed description and accompanying drawings. U. S. RE37, 793, U.S.A. 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 and Non-Patent Publications referenced herein and / or listed in application data sheets, including US Patent No. 6,380,248. All of the items are individually incorporated herein by reference as if individually incorporated. Aspects and embodiments of the present invention may be modified as needed, and various patents, applications and publication concepts may be used to provide further embodiments.

Brief description of multiple appearances of the drawing
Pseudomonas aeruginosa colony biofilm viability (logCFU; colony forming units) obtained after growing for 24 hours at 37 ° C. on 10% trypsin soy agar (TSA) for 18 hours following the indicated treatment Is shown. The indicated antibiotic treatments were 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, the lowest concentration that prevents bacterial growth). Shown are the viable counts (log CFU) of Staphylococcus aureus colony biofilms obtained after growing for 24 hours on 10% trypsin soy agar and then performing the indicated treatments. The indicated antibiotic treatments 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). FIG. 4 shows the wound closure over time of keratinocytes exposed to a biofilm. (*) Significant difference from control (P <0.001). Figure 3 shows sub-inhibited BisEDT that suppresses antibiotic resistance to multiple antibiotics. Figure 2 shows the effect of antibiotics with and without BisEDT on the flora of MRSA (methicillin-resistant Staphylococcus aureus). Panel A shows only a standard antibiotic soak disc. [GM = gentamicin, CZ = cefazolin, FEP = cefepime, IPM = imipenem, SAM = ampicillin / sulbactam], LVX = levofloxacin. Figure 3 shows sub-inhibited BisEDT that suppresses antibiotic resistance to multiple antibiotics. Figure 2 shows the effect of antibiotics with and without BisEDT on the flora of MRSA (methicillin-resistant Staphylococcus aureus). Panel B shows a disc mixed with BisEDT (BE). [GM = gentamicin, CZ = cefazolin, 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 mean variability of BPC (2-fold serial dilution step) with 0.25 μM BisEDT (n = 3). S. cerevisiae grown for 48 hours at 37 ° C. in TSB + 2% glucose. Figure 2 shows the effect of BisEDT and antibiotics on the development of Epidermidis. Results are expressed as the mean MIC (dilution step) with increasing BisEDT (n = 3). See the legend in FIG. 5 for definitions of antibiotics. In bone and metals samples of open fractures in an in vivo rat model with and without cefazolin antibiotic treatment, in addition to the three BT formulations, Bis-EDT, MB-11 and MB-8-2 5 is a bar graph showing the average levels of S. aureus detected. The standard error of the average value is shown as an error bar. Early euthanized animals 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 directed to certain bismuth-thiol (BT) compounds provided herein (preferably BT having a volume average diameter of about 0.4 μm to about 5 μm). (Including microparticles) is a powerful disinfectant, anti-bacterial and / or anti-biofilm against certain bacteria, including those associated with a number of clinically significant infections, including infections that may include bacterial biofilms It is based on the surprising discovery that it shows activity but not other specific BT compounds (even when provided as microparticles).

  Contrary to expectation, not all BT compounds are uniformly effective against such bacteria in a predictable manner, but instead exhibit different potencies depending on the target bacterial species. In particular, as described herein, specific BT compounds can be applied in a manner that for the first time can provide clinically relevant strategies for the management of bacterial infections, including bacterial biofilm infections, by non-limiting theory. (Preferably containing BT microparticles having a volume average diameter of about 0.4 μm to about 5 μm) have been found to show higher efficacy against Gram-negative bacteria, but other specific BT compounds (preferably Containing BT microparticles having a volume average diameter of about 0.4 μm to about 5 μm) were 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 a particle size (eg, from about 0.4 μm to about 5 μm) as disclosed herein. It relates to the surprising advantages provided by the novel bismuth-thiol (BT) compositions that can be produced in preparations 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 as disclosed herein, for certain embodiments, certain antibiotic antimicrobial and anti-biofilms previously found to lack a strong therapeutic effect against such bacterial infections. Efficacy may be achieved by treating the infectious agent with one or more of these antibiotics and the selected BT compound in any order, cooperatively, simultaneously, or sequentially (eg, at the site of infection, such as a natural surface). (Directly applied to the surface or interior of a) can be significantly enhanced (eg, 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 were identified herein in combination with certain antibiotics for antibacterial and / or anti-biofilm activity against certain strains or strains. The provided synergistic or augmenting combinations can be provided. Certain BT / antibiotic combinations act synergistically or enhance potency against certain bacteria, while other specific BT / antibiotic combinations may provide such synergistic or potent antimicrobial and The observation that no anti-biofilm activity could be demonstrated demonstrates the unexpected properties of such a combination, described in more detail below.

  According to these and related embodiments, the antibiotic and BT compound may be administered simultaneously or sequentially, in any order, and the particular infectious agent (eg, formed by gram-negative or gram-positive bacteria). A specific synergistic or potent combination of one or more of the antibiotics disclosed herein and one or more BT compounds to treat a treated biofilm) is predictable (eg, Exhibit no additive) activity, but instead, unexpectedly synergistic or potentiating (eg, superphase) as a function of the selected antibiotic, the selected BT compound and the specifically identified target bacterium. It is noteworthy that it worked in an (additive) manner.

  For example, but not by way of limitation, in the context of a wide variety of natural and / or artificial surfaces that are actually or potentially microbial infected, and furthermore, the aspects of improved substantially monodisperse particulate BT formulations In certain embodiments, either or both of certain antibiotic compounds and certain BT compounds disclosed herein may exert a slight antimicrobial effect when used alone against certain strains or strains However, the combination of both the antibiotic compound and the BT compound showed a greater magnitude (statistically significant difference) than the simple sum of the effects of each compound when used alone against the same strain or strain. Exerts a potent antimicrobial effect, thereby, according to non-limiting theory, anti-BT and / or anti-BT effects on antibiotic efficacy. 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 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 an antibiotic and a BT compound are, surprisingly, naturally and / or artificially described herein. It confers strong antibacterial effects on certain bacteria, such as antimicrobial effects on biofilms formed by certain bacteria in certain environments, such as surfaces, and even in certain situations.

  That is, certain BTs, including 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, such synergism may be due to the presence of the antibiotic but no BT compound and / or the presence of the BT compound but no antibiotic In some cases, it will manifest as a detectable effect on a larger scale than the effect that can be detected (ie, in a statistically significant manner with respect to appropriate control conditions). Similarly, certain BT-antibiotic combinations show potentiating (0.5 <FICI <1.0), such potentiating when an antibiotic is present but no BT compound is present, and And / or in the presence of a BT compound but no antibiotic, manifest as a detectable effect on a larger scale than the effect that can be detected (ie, in a statistically significant manner with appropriate control conditions).

  Examples of such detectable effects include, in certain embodiments, (i) preventing infection by a bacterial pathogen, (ii) inhibiting cell viability or cell growth 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 development of substantially all biofilm-type cells of bacterial pathogens, wherein the present invention provides Without limitation, in other contemplated embodiments, the antibiotic-BT synergy is one or more other clinically important parameters, such as one or more antibiotics or other drugs or chemicals. Degree of resistance or susceptibility of the bacterial pathogen to the agent, specific to one or more chemical, physical or mechanical conditions (eg, pH, ionic strength, temperature, pressure) The degree of resistance and susceptibility of the pathogen and / or one or more biological agents (eg, a virus, another bacterium, a biologically active polynucleotide, an immune cell or immune cell product, such as an antibody, a cytokine, a chemokine, a degrading enzyme; One or more species that can include a change (eg, a statistically significant increase or decrease) in the degree of resistance or susceptibility of the bacterial pathogen to the starting enzyme, membrane disrupted proteins, free radicals, such as reactive oxygen species, and the like. It may be expressed as a detectable effect.

  As provided herein, antibiotics present when the effect of a synergistic or potent 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 may determine the effect of a particular agent on structure, function and / or activity of a bacterial population for the purpose of ascertaining substance-BT synergy or potentiation. Wax (for example, Coico et al. (Eds.), Current Protocols in Microbiology, 2008, John Wiley & Sons, Hoboken, NJ; 007, CRC Press, Boca Raton, FL).

  For example, in certain embodiments, antimicrobial effects, as described herein, are measured using various concentrations of candidate agents (eg, BT and antibiotics, alone or in combination) to determine the concentrations of Eliopoulos, et al. (Antibiotics). in Laboratory Medicine (Lorian, V., Ed.), pp. 330-96, Williams and Willkins, Baltimore, MD, Elliopoulos and Moelling, part of the Ellipouls and Moelling index (1996), in the USA By calculating the germicidal concentration index (FBCI), synergy may be determined. Synergy may be defined as less than or equal to 0.5 and antagonism as a FICI or FBCI index greater than 4 (eg, Odds, FC (2003) Synergy, antagonism, and what the chequerboard jours betwe hen. Antimicrobial Chemotherapy 52: 1). Synergy is conventionally defined as a four-fold or more reduction in antibiotic concentration, or using a 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, an antibiotic and a BT composition) and is greater than when the concentration of the second drug is replaced with the first drug ( For example, it may be defined as the effect of the combination (in a statistically significant manner).

  Thus, as described herein, in certain preferred embodiments, the combination of BT and antibiotic will be understood to be synergistic if a FICI value of 0.05 or less is observed (Odds, 2003). As also described herein, in certain other preferred embodiments, and according to non-limiting theory, certain BT-antibiotic combinations exhibit such high synergistic potential. It is disclosed that it may exhibit a FICI value of 0.5 to 1.0, the synergy of which is the non-optimal of at least one BT and at least one antibiotic showing unilateral or mutually enhanced cooperative antimicrobial efficacy May be observed using concentration. Such an effect may be referred to herein as "enhancing" antibiotic activity or "enhancing" BT activity.

  According to certain embodiments, (i) a concentration (in a statistically significant manner) lower than the characteristic minimum inhibitory concentration (MIC) of BT for a given target microorganism (eg, a given strain or strain). And at least one BT of (ii) lower than the characteristic IC50 (a concentration that inhibits the growth of 50% of the microbial population; for example, Soothill et al., 1992 J Antimicrob Chemother 29 (2): 137) (statistics). And / or 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. If an antimicrobial (eg, BT or antibiotic) is used at the same concentration in the absence of the other antimicrobial (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 a preferred embodiment, "enhancing" 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, the synergistic or potentiating antibiotic and / or BT activity is increased by methods known in the art, such as by adding Loewe to Based models (eg, FIC index, Greco model), or Bliss independence based models (eg, non-parametric and semi-parametric models) or other methods known in the art described herein (eg, Meliadias et al., 2005 Medical Mycology 43: 133-152). Exemplary methods for measuring synergistic or potentiating antibiotic and / or BT activity in this manner are described, for example, in Meliadias et al. , 2005 Medical Mycology 43: 133-152 and the references cited therein (also, Meliadias et al., 2002 Rev Med Microbial 13: 101-117; White et al., 1996 Antimicrobage Agency: 1996). Mouton et al., 1999 Antimicrob Agents Chemother 43: 2473-1478).

  Another specific embodiment is that BT compounds and antibiotics do not show predictable (simply additive) activity in vivo, but instead use selected antibiotics, selected BT compounds, and infectious agents. Unexpectedly synergistic or augmenting (eg, super-additive; or when 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 agent were replaced with the first agent, One or more antibiotics disclosed herein that may show synergistic or potentiating effects 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. Thus, according to these and related embodiments, in certain in vivo situations, FICI or FBCI values (measured in vitro) may not be immediately available, but instead a synergistic or potentiating effect of the BT-antibiotic. It will be appreciated that may be determined in a manner that results from a quantifiable measure of the infectious agent.

  For example, in one embodiment such as the rat murine femoral critical defect model of open fracture in vivo described in Example 11, observed after treatment with BT-antibiotic combination compared to antibiotic treatment or BT compound alone. A statistically significant decrease in bacterial count is indicative of a synergistic or enhancing 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 lesion after treatment with the BT-antibiotic combination compared to the antibiotic treatment or BT compound alone. A decrease of 40%, or 50%, observed in this or other in vivo models is considered an indicator of a synergistic or potentiating effect.

  Other exemplary indicators of in vivo infection include established methodologies developed to quantify the severity of the infection, such as various wound scoring systems known to those of skill in the art (eg, the European Wound Management Association (EWMA), Position Document: Identifying Criterion 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 ASESIS (Wilson AP, J Hosp Infect 1995; 29 (2)). : 81-86; Wilson et al., Lancet 1986; . See Horan TC, Gaynes P, Martine WJ, et al. , 1992 Infect Control Hosp Epidemiol 1992; 13: 606-08. In addition, recognized clinical indicators of wound healing known to the skilled clinician, such as wound size, depth, granulation tissue status, infection, etc., in the presence or absence of BT compounds and / or antibiotics May be measured. Thus, based on the present disclosure, to determine whether a BT composition-antibiotic combination alters in vivo wound healing (eg, increases or decreases in a statistically significant manner with respect to appropriate controls). Various methods will be readily apparent to those skilled in the art.

  In view of these and related embodiments, microbial-infected natural and artificial surfaces, such as those 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 (eg, in certain embodiments, a therapeutically effective amount) of a composition or formulation comprising one or more synergistic antibiotics or one or more potent antibiotics provided herein. A great variety of methods for treating with are provided herein. Based on the present disclosure, it will be appreciated that certain antibiotics previously deemed ineffective by a person of ordinary skill in the art against a given type of infection are now contemplated for use in treating the same type of infection. I want to.

  Thus, certain embodiments contemplate compositions comprising one or more BT compounds used as disinfectants. Disinfectants are substances that kill or impede the growth of microorganisms, and are typically applied to living tissue to distinguish them from fungicides that are normally applied to inanimate subjects. (Goodman and Gilman's "The Pharmacological Basis of Therapeutics", Seventh Edition, Gilman et al., Editors, 1985, and in Gandh mad. Common examples of disinfectants are ethyl alcohol and tincture of iodine. Disinfectants include disinfectants that kill microorganisms, such as microbial pathogens.

  Certain embodiments described herein use one or more BT compounds and one or more antibiotic compounds (eg, synergistic and / or potentiating antibiotics provided herein). Compositions comprising the same may be contemplated. Antibiotics are known in the art and are typically produced by one species of microorganism and produced from a compound that kills another species of microorganism, or the same or similar chemical structure and mechanism of action. Such as drugs that destroy microorganisms in or on living organisms when applied topically. In particular, the 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 penicillins, and aminopenicillins. That is, the antibiotics need not be limited, but include, but are not limited to, oxacillin, piperacillin, cefuroxime, cefotaxime, cefepime, imipenem, aztreonam, streptomycin, tobramycin, tetracycline, minocycline, ciprofloxacin, levofloxacin, erythromycin, linezolid, fosfomycin, fosfomycin, and capresiomycin. , Isoniazid, ansamycin, carbacephem, monobactam, nitrofuran, penicillin, quinolone, sulfonamide, clofazimine, dapsone, capreomycin, cycloserine, ethambutol, ethionamide, isoniazid, pyrazinamide, rifampicin, rifampin, rifabutin, rifapentine, streptomycin, streptomycin, streptomycin Namin, chloramphenicol, phospho Isine, fusidic acid, linezolid, metronidazole, mupirocin, platencimycin, quinupristin, dalfopristin, rifaximin, thiamphenicol, tinidazole, aminoglycoside, β-lactam, penicillin, cephalosporin, carbapenem, fluroquinolone, ketolide, lincosamide, lincosamide Oxazolidinone, streptogramin, sulfonamide, tetracycline, glycylcycline, methicillin, vancomycin, nafcillin, gentamicin, ampicillin, chloramphenicol, doxycycline, tobramycin, amikacin, arbekacin, gentamicin, kanamycin, neomycin, netilmycin, paromomycin, rhodomycin, rhodomycin, rhodromycin Streptomycin, Tobrama Mention may be made of isin, 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 (e.g., Washington University School of Medicine, The Washington American Manual of Medical Therapeutics, London, 2000, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd, 32nd to 32th, and 30th). and Wilkins, Philadelphia, PA; Hauser, AL, Antibiotic Basics for Clinicians, 2007 Lippincott, Williams and Wilkins, Philadelphia, PA).

  An exemplary class of antibiotics for use with one or more BT compounds in certain embodiments disclosed herein is Edson RS, Terrell CL. The aminoglycosides. Mayo Clin Proc. Aminoglycosides, reviewed in 1999 May; 74 (5): 519-28. This class of antibiotics inhibits bacterial growth by damaging bacterial protein synthesis through the binding and inactivation of bacterial ribosomal subunits. Aminoglycosides, in addition to such bacteriostatic properties, also have a bactericidal effect through the disruption of the cell wall of Gram-negative bacteria.

  Aminoglycoside antibiotics include gentamicin, amikacin, streptomycin and the like, which are generally considered useful for the treatment of Gram-negative bacteria, mycobacteria and other microbial pathogens, but classification of resistant strains has been reported. Aminoglycosides are not absorbed from the gastrointestinal tract and are therefore not generally 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 for treating epithelial tissue surfaces at one or more locations, for example, along the oral cavity, gastrointestinal tract / gastrointestinal tract. Oral administration of the BT / antibiotic combination is contemplated (eg, the antibiotic need not be limited to aminoglycosides). Also contemplated in other specific embodiments are herein described as fungicides, which refer to preparations that kill or block the growth of microorganisms on the outer surface of an inanimate subject. The use of the compositions and methods provided.

  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.A. S RE37,793, U.S. Pat. S. No. 6,248,371, 6,086,921, and 6,380,248 (for example, see also US Pat. No. 6,582,719) (including the preparation method). It may be a composition comprising, initially, bismuth or a bismuth salt and a thiol (eg, -SH, or sulfhydryl) containing compound. However, certain embodiments are not so limited, and may contemplate other BT compounds, including bismuth or bismuth salts and thiol-containing compounds. The thiol-containing compound may contain 1, 2, 3, 4, 5, 6, or more thiol (eg, -SH) groups. In a preferred embodiment, the BT compound comprises bismuth associated with a thiol-containing compound via an ionic bond and / or as a coordination complex, although in some other embodiments, the bismuth is contained in May be associated with a thiol-containing compound via a covalent bond, as may be found in However, certain contemplated embodiments explicitly exclude organometallic compounds, such as BT compounds, where bismuth is a compound found in a covalent bond 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 to a single bismuth atom is shown for comparison. Atomic ratios shown may not be the exact molecular formulas of all species in a given preparation. The number in parentheses is the ratio of bismuth to one (or more) thiol agent (eg, Bi: thiol1 / thiol2). "CPD": Compound.

  The BT compounds used in certain of the embodiments disclosed herein may be prepared according to established procedures (eg, US RE 37,793, US 6,248,371, 6,086, Nos. 921 and 6,380,248; Domenico et al., 1997 Antimicrob. Agent. Chemother. 41 (8): 1697-1703, Domenico et al., 2001 Antimicrob. Agent. Chem. 14117-1421), and in certain other embodiments, the BT compound may be prepared according to the methodology described herein. In other words, certain preferred embodiments are those which have sufficient conditions and times for the formation of a precipitate comprising microparticles comprising a BT compound (e.g., a concentration, solvent described herein and understood by one of ordinary skill in the art based on the present disclosure). 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 500 mM, at least 50 mM, at least An aqueous solution of acidic bismuth containing at a concentration of 600 mM, at least 700 mM, at least 800 mM, at least 900 mM, or at least 1 M and free of a hydrophilic, polar or organic solubilizer is mixed with ethanol to form a first ethanol solution. Obtaining a reaction solution by obtaining a solution and reacting 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, BT The synthetic methods described herein for preparing the compounds, particularly for obtaining the BT compounds in substantially monodispersed 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); 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).

  Without wishing to be bound by theory, certain preferred embodiments may include 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 compounds can be performed, if desired, with ease of mass production, improved product purity, homogeneity or consistency (including particle size uniformity), or topical formulations of the present invention. It is believed that a composition comprising the BT compound can be obtained having one or more desired properties, including other properties useful for the preparation and / or administration of the BT compound.

  In certain embodiments, the BT compositions prepared by the methods described herein for the first time have, according to certain currently preferred embodiments, each a volume average diameter (VMD) of about 0.4 μm to about 5 μm. It has been discovered that it exhibits advantageous homogeneity in connection with its appearance as a substantially monodisperse particulate suspension. The particle size measurement can be referred to as the volume mean diameter (VMD), median mass diameter (MMD), or median aerodynamic 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, such as standard humidity, are maintained. However, if humidity is not maintained, dehydration during impactor measurements will result in lower MMD and MMAD measurements than VMD. For purposes of this specification, VMD, MMD and MMAD measurements are considered to be under standard conditions, so the descriptions of VMD, MMD and MMAD will be equivalent. Similarly, particle size measurements of dry powders in MMD and MMAD are considered equivalent.

  As described herein, preferred embodiments relate to a substantially monodispersed suspension of BT-containing microparticles. Generating a defined BT particle size with a small geometric standard deviation (GSD) may include, for example, BT attachment, accessibility to a desired target site within or on a natural surface, and / or BT microparticles May be optimized for tolerability by the subject to which it is administered. A narrow GSD will limit the number of particles outside the desired VMD or MMAD size range.

  In one embodiment, there is provided a liquid or aerosol suspension of microparticles having a VMD of about 0.5 microns to about 5 microns containing one or more BT compounds disclosed herein. In another embodiment, a liquid or aerosol suspension having a VMD or MMAD from about 0.7 microns to about 4.0 microns is provided. In another embodiment, a liquid or aerosol suspension having a VMD or MMAD from about 1.0 micron to about 3.0 microns is provided. In other specific embodiments, the VMD is from 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 0.5. 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 as described herein. A liquid suspension containing one or more BT compound particles prepared as described above is provided.

  Thus, in certain preferred embodiments, a "substantially" monodisperse BT preparation described herein for the first time, e.g., "substantially" all of the microparticles is within a specific range (e.g., about 0%). A BT composition comprising a BT compound in particulate form having a volume average diameter (VMD) of about 0.4 μm to about 5 μm) comprises 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 a VMD within the cited size range.

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

  In addition, or alternatively, the substantially monodisperse BT microparticles described herein may be advantageously used without the need for micronization, ie, expensive and labor-intensive milling or supercritical fluid processing, or microparticles. May be manufactured without the other equipment and procedures typically used to produce (e.g., Martin et al. 2008 Adv. Drug Deliv. Rev. 60 (3): 339; Morive et al. 60, (3): 328; Cape et al., 2008 Pharm. Res. 25 (9): 1967; Rasenack et al., 2004 Pharm. Dev. Technol .: 9 (1)., 2008 Adv. 1-13). Thus, embodiments of the present invention may include, without limitation, enhanced and substantially homogeneous solubilizing properties, suitability for desired dosage forms such as oral, inhalation or topical forms for dermatological / dermal wounds, It offers the beneficial effects of a substantially homogeneous microparticle preparation, including a high degree of bioavailability and other beneficial properties.

  The particulate suspension of the BT compound may be a wetting agent, surfactant, mineral oil, or other ingredients or additives as would be known to those skilled in the formulation, for example, to retain individual particulates in the suspension. Is administered as an aqueous formulation, such as a preparation containing, as a suspension or solution in aqueous and organic solvents, 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 using, for example, either hydraulic or ultrasonic spraying. Propellant-based systems may use a suitable pressurized dispenser. As the dry powder, a dry powder dispersion device that can effectively disperse the BT-containing fine particles may be used. By selecting the appropriate device, the desired particle size and distribution may be obtained.

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

  Under general conditions, the method comprises: (a) under conditions and for a time sufficient to obtain a solution substantially free of solid precipitates, comprising (i) a bismuth salt comprising a bismuth at a concentration of at least 50 mM; An acidic aqueous solution without a polar, polar or organic solubilizer, (ii) admixed with a sufficient amount of ethanol to form at least about 5%, 10%, 15%, 20%, 25% by volume. Or obtaining a mixture comprising 30% by volume, preferably about 25% by volume of ethanol; and (b) from about 1: 3 to bismuth under conditions and for a time sufficient to form a precipitate containing the BT compound. Adding an ethanolic solution containing the thiol-containing compound present in the reaction solution in a molar ratio of about 3: 1 to the mixture of (a) to obtain a reaction solution.

  In certain preferred embodiments, the bismuth salt may be Bi (NO3) 3, but it will be understood by the present disclosure that bismuth may be provided in other forms. In certain embodiments, the concentration of bismuth 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%, 10%, 15%, 20%, 22%, or 22.5% by weight of bismuth. The acidic aqueous solution may comprise at least 5% by weight or more nitric acid in certain preferred embodiments, and in other specific embodiments, the acidic aqueous solution comprises at least 0.5% by weight, at least 1% by weight. At least 1.5%, at least 2%, at least 2.5%, at least 3%, at least 3.5%, at least 4%, at least 4.5% or at least 5% by weight of 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, Containing one or more of 3,4-dimercaptotoluene, 2,3-butanedithiol, 1,3-propanedithiol, 2-hydroxypropanethiol, 1-mercapto-2-propanol, and dithiothreitol Is 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 [2C3mercaptan]), butanethiol (C4H9SH [n-butylmercaptan]), tert-butylmercaptan (C (CH3) 3SH [t-butylmercaptan]), pentanethiol (C5H11SH [pentyl mercaptan]), coenzyme A, lipoamide, glutathione, cysteine, cystine, 2-mercaptoethanol, dithiothreitol, dithioerythritol, 2-mercaptoindole, transglutaminase, 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-butanedithiol diacetate, 1,5- Pentanedithiol, 1,6-hexanedithiol, 1,8-octanedithiol All, 1,9-nonanedithiol, adamantanethiol, 1-butanethiol, 1-decanethiol, 1-dodecanethiol, 1-heptanethiol, 1-heptanethiol plum, 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-tetradecanethiol plum, 1-undecanethiol, 11- (1H-pyrrol-1-yl) undecane-1-thiol, 11-amino-1 -Un Canthiol hydrochloride, 11-bromo-1-undecanethiol, 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-hexanethiol plum, 3- (dimethoxymeth Lucylyl) -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-nonanoe , 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 ™ 8, NanoThinks ™ ACID11, NanoThinks ™ ACID16, NanoThinks ™ ALCO11, NanoThinks ™ THIO8, Octanthio- 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- Thiols, p-terphenyl-4,4 "-dithiol, tert-dodecylmercaptan and tert-nonylmercaptan.

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

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

  In this context, the method of the present invention provides that bismuth is not water soluble at 50 μM (eg, US RE37793) and bismuth is unstable in water (eg, Kuvshinova et al., 2009 Russ. J. Inorg. Chem 54 (11): 1816), surprising and anticipated in view of the generally recognized art that bismuth is also unstable in aqueous nitric acid without the presence of hydrophilic, polar or organic solubilizers. Present outside benefits. For example, in all of the most trusted descriptions of BT preparation methodology (eg, Domenico et al., 1997 Antimicrob. Agents. Chemother. 41: 1697; US. 6,380,248; US RE37, 793). No. 6,248,371), the hydrophilic solubilizer propylene glycol is required to dissolve the bismuth nitrate, and the bismuth concentration of the solution prepared for reaction with thiol is 15 mM. Much lower than that, which limits the available modes of production of BT compounds.

  In contrast, the present disclosure does not require a hydrophilic, polar or organic solubilizer to dissolve bismuth, and surprisingly achieves higher concentrations. 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 agents. Other highly polar water-miscible organic materials include dimethylsulfoxide (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, Wychch (Ed.), (See also Catalan, 2001 in Andrew Publ., NY, and references cited therein), which may be selected based on solvent polarity / polarizability (SPP) scale values, 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 determining the SPP value of a solvent based on UV measurements of 2-N, N-dimethyl-7-nitrofluorene / 2-fluoro-7-nitrofluorene probe / homomorph pair has been described ( Catalan et al., 1995).

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

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

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

  Briefly, by way of non-limiting example, an excess of room temperature 5% aqueous HNO3 (11.4 L) may be added to a Bi (NO3) 3 solution (eg, 43% Bi (NO3) 3 (w / w), 5%). % Aqueous nitric acid (w / w), 52% water (w / w), 0.331 L (about 0.575 mol) of an acidic bismuth aqueous solution such as water 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 to give 1,2-ethanedithiol [〜0 .55M], an ethanolic solution (1.56 L) of a thiol compound 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 slowly 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 formed product precipitates as a precipitate containing the particulates described herein, and is then collected by filtration and subsequently washed with ethanol, water and acetone to give BisEDT as a yellow amorphous powder solid. obtain. The crude product may be redissolved in anhydrous alcohol with stirring, then filtered and washed successively several times with ethanol and then several times with acetone. The washed powder may be triturated in 1 M NaOH (500 mL), filtered and subsequently washed with water, ethanol and acetone to obtain purified particulate BisEDT.

  According to 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 assimilation of EPS can contribute to bacterial virulence, such as impairing wound healing. However, bismuth alone is not therapeutically useful as an intervention and is instead typically administered as part of a complex such as BT. That is, bismuth-thiol (BT) is a family of compositions comprising compounds derived from the chelation of bismuth with thiol compounds and exhibiting a dramatic improvement in the antibacterial therapeutic efficacy of bismuth. BT shows significant anti-infective, anti-biofilm, and immunomodulatory effects. 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 (subinhibitory) concentrations, prevent many pathogenic features of common wound pathogens at the same subinhibitory level, and prevent septic shock in animal models, Can be synergistic with many antibiotics.

  As described herein, such synergy in the antimicrobial effect of one or more specific BTs when combined with one or more specific antibiotic compounds is due to the distinct antibiotics against a particular bacterial type. And not immediately predictable based on the profile of the BT effect, but surprisingly, specific considerations for specific bacterial populations, such as confirmation of the presence of Gram-negative or Gram-positive bacteria (or both) It may be obtained by selecting a BT-antibiotic combination. For example, as disclosed herein, certain BT synergistic antibiotics include amikacin, ampicillin, aztreonam, cefazolin, cefepime, chloramphenicol, ciprofloxacin, clindamycin (or other lincosamide antibiotics). Substances), daptomycin (Cubicin®), doxycycline, gatifloxacin, gentamicin, imipenim, levofloxacin, linezolid (Zyvox®), minocycline, nafcillin, paromomycin, rifampin, sulfamethoxazole, tetracycline, tobramycin And one or more of vancomycin. For example, MRSA that has little or no sensitivity individually to gentamicin, cefazolin, cefepime, sphamethoxazole, imipenim or levofloxacin, when exposed to these antibiotics in the presence of the BT compound BisEDT, will cause any of the antibiotics In vitro tests have shown that they show significant susceptibility to S. aureus. Thus, certain 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, rifampin, sulfamethoxazole, tobramycin and vancomycin. Explicitly contemplated compositions and / or methods that may include a combination with one or more antibiotics, other specific embodiments contemplated herein include a BT compound and amikacin, Picilin, aztreonam, cefazolin, cefepime, chloramphenicol, ciprofloxacin, clindamycin (or other lincosamides), daptomycin (Cubicin®), doxycycline, gatifloxacin, gentamicin, imipenim, levofloxacin, One or more antibiotics, wherein one or more antibiotics selected from linezolid (Zyvox®), minocycline, nafcillin, paromomycin, rifampin, sulfamethoxazole, tobramycin and vancomycin can be explicitly excluded; Compositions and / or methods that may include combinations of are contemplated. It should be noted herein that gentamicin and tobramycin belong to the aminoglycosides of antibiotics. 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; Veloila 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. Pharmaceut. 373: 141-146, it is noted that none of these publications teach or suggest the monodisperse particulate BT compositions disclosed herein. I want to.

  Thus, as described herein, in certain preferred embodiments, comprising the particulate BT described herein, and optionally, in other specific embodiments, synergistic and / or potentiating antibiotics as well. Compositions and methods for treating a plant, animal or human subject, or product with a composition comprising the same are provided. Those of skill in the relevant art will recognize, based on the present disclosure, appropriate agricultural, clinical, commercial, industrial, manufacturing, domestic, and other aspects and situations in which such treatment may be desirable. The criteria include, inter alia, geriatric, cardiovascular, metabolic disorders (eg, diabetes, obesity, etc.), infections and inflammation (airway or (E.g., the epithelial lining of the gastrointestinal tract, or other epithelial surfaces such as glandular tissue), and the medical arts, including related medical and subspecialties.

  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, the bismuth-thiol (BT) described herein. A composition comprising a compound may include a compound that, in certain different 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 of making them are disclosed herein and are described, for example, in Domenico et al. (1997 Antimicrob. Agent. Chemother. 41 (8): 1697-1703; 2001 Antimicrob. Agent. Chemother. 45 (5): 1417-1421) and U.S. Pat. S RE37,793, U.S. Pat. S. Nos. 6,248,371, 6,086,921, and 6,380,248. As also mentioned earlier, certain preferred BT compounds include those containing a bismuth or bismuth salt in a coordination complex with or with a thiol-containing compound, such as bismuth chelated to a thiol-containing compound. Other particularly preferred BT compounds are those that contain bismuth or a bismuth salt in a covalent link to the thiol-containing compound. Also preferred are the substantially monodisperse particulate BT compositions described herein. For the first time described herein, there have been uses from previous efforts to treat bacterial infections or in compositions and methods for promoting the herein described treatment of natural and / or artificial surfaces. The characterization of other compounds in other aspects could 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 at least one particulate BT compound as described herein to a natural surface. In certain embodiments, the method may be, in certain preferred embodiments, a synergistic antibiotic described herein, and in certain other embodiments, described herein. The method further includes administering the at least one antibiotic compound, which may be a potentiating antibiotic, simultaneously or sequentially in any 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 herein, e.g., Washington University School of Medicine, The Washington American Manual of Medical Therapeutics, 32nd Ed., Therapeutics, 32nd Ed. , PA; Hauser, AL, Antibiotic Basics for Clinicians, 2007 Lippincott, Williams and Wilkins, Philadelphia, PA.

  As described herein, certain embodiments provide that in the case of bacterial infections, including Gram-positive bacteria, a preferred therapeutically effective formulation is a BT compound (eg, BisEDT, bismuth; 1,2- BisPyr, bismuth: pyrithione; BisEDT / Pyr, bismuth: 1,2-ethanedithiol / pyrithione) and rifamycin, or a BT compound and daptomycin (Cubicin®, Cubis Pharmaceuticals, Lxinton, Mass.), 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, gatifuro The unexpected finding that it can include one or more of the following: oxacin, gentamicin, imipenim, levofloxacin, linezolid (Zyvox®), nafcillin, paromomycin, rifampin, sulfamethoxazole, tobramycin and vancomycin. Comes 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 can include a BT compound and amikacin. Derived from discovery. Certain related embodiments contemplate treating an infection 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. Accordingly, in view of these embodiments, other related embodiments are directed to the field of medical methodology as selecting appropriate antibiotic compounds for inclusion in formulations 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 on or on natural or artificial surfaces by well-known criteria of being Gram-positive or Gram-negative.

  The compositions and methods described herein typically employ compounds described herein (eg, with only one or more particulate BT alone, or with one or more of the synergistic compounds disclosed herein). And / or in combination with potentiating antibiotics), applied or administered to a microbial site, such as the presence of a microbe on or within a natural or artificial surface, to provide a microbe in various aspects (eg, bacteria, viruses, yeasts, molds). And other fungi, microbial parasites, etc.). Such natural surfaces include, but are not limited to, all surfaces of plants (eg, roots, bulbs, stems, leaves, branches, vines, creeping branches, buds, flowers or parts thereof, shoots, fruits, seeds, pods, etc. Or parts thereof, mammalian tissues (eg, skin, scalp, lining of the digestive tract, epithelium such as oral cavity; endothelium, cells and tissue membranes such as peritoneum, pericardium, pleura, periosteum, meninges, myofibers membrane, etc .; cornea) And other mammalian tissues, such as muscle, heart, lung, kidney, liver, spleen, gallbladder, pancreas, bladder, nerves, teeth, bones, joints, tendons, ligaments, etc. Any part of the product where the presence of microorganisms can be found, such as walls, windows, floors, underfloor spaces, attics, basements, fences, roofs of commercial, residential, industrial, educational, health care and other public buildings , Ceilings, lighting and plumbing fixtures, vents, ducts, pipelines, doorknobs, Switches, sanitary equipment, drainage equipment, tanks, water pipes; medical and dental equipment, implants, tools, instruments, equipment, etc .; metal, glass, plastic, wood, rubber and paper products; transport equipment, such as shipping containers, automobiles, Railway equipment, boats, ships (eg, external hulls, rudders, anchors and / or propeller surfaces, internal fixtures and ballast tanks, and other internal surfaces), barges, and other marine equipment, such as docks, bulkheads, piers, etc. Are also mentioned.

  The particulate antimicrobial agents described herein can be used to treat products, including natural and / or artificial surfaces, to inhibit microbial growth, reduce microbial infection, Improve biofilms, prevent bacterial to biofilm conversion, prevent or inhibit microbial infection, prevent damage, and any other described herein. May be used for use. These agents include multiple anti-viral agents, including the prevention or inhibition of viral infection by herpes family viruses, such as cytomegalovirus, herpes simplex type 1 and herpes simplex type 2, and / or infection by other viruses. It is also useful for the purpose. In this context, the agent may be a variety of viruses, such as single-stranded RNA virus, single-stranded DNA virus, 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 roll virus; potato virus A, M, S, X or Y; tomato yellow gangrene virus; grape roll associated virus 3; plum Pox virus; 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 The treatment or treatment of Helicobacter pylori infections and genus (eg Candida albicans, Candida glabrata, C. parapsilosis, C. tropicalis, and C. dubliniensis) or Helicobacter pylori infection and peptic ulcer disease In one embodiment, the agent is administered at a dosage that is not generally lethal to bacteria, but is still sufficient to reduce protective polysaccharide coatings that otherwise resist the natural immune response. This technique is thus used in human symbiotic microorganisms (eg, normal intestinal flora). By way of example and not by way of limitation, certain contemplated embodiments are described herein, which will assist in eradicating a bacterial infection through the immune system to the extent that it would become a case using antibiotics without damage. .

  Particulate Bismuth-Thiol for Coating and Treating Flutes In one embodiment, preventing and / or controlling (ie, slowing, slowing, inhibiting) biofilm development, disrupting biofilms, Or a water pipe (eg, a water pipe used by dentists, dental hygienists and other oral care professionals and caregivers), or other water delivery vehicles, such as pipes, pipes, faucets, fountains, showerheads, or humans Or any other instrument or device that comes into contact with or delivers water consumed by or applied to a non-human animal (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 a biofilm on an outer surface. These methods can also be useful for preventing, reducing, suppressing, eliminating or preventing 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.

  Biofilms are a microscopic community composed 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 coolant and cleaning agent in dental procedures can be heavily contaminated by microorganisms (see, for example, the Environmental Protection Agency website: epa.gov/safewater/mcl/html). Pathogenic microorganisms or opportunistic pathogens found in water from dental water pipes and instruments include actinomycetes, Bacteroides, Bacillus, Cropsporidium, Escherichia, Flavobacterium, Klebsiella, Legionella. Genus, Moraxella, Mycobacterium, Peptostreptococcus, Pseudomonas, Staphylococcus, Streptococcus, and Bayonella. In addition, as a result of biofilm formation, Legionella species and protozoa can grow in water pipes or water delivery vehicles. Bacteria from the biofilm as well as other microorganisms present in the water conduit or water delivery vehicle are continuously released as a stream of water through the water conduit 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 / cleanser for non-surgical dental procedures is based on the number of aerobic heterotrophic plates (HPC). Is ≦ 500 CFU / ml. The American Dental Association (ADA) has put forward even more stringent standards, recommending that water used for dental procedures contain bacterial levels of <200 CFU / ml. Measurements made to keep bacterial counts in dental water systems at low levels include the use of antimicrobial agents (see, e.g., 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; water pipes and water. Use of chemicals as disinfectants (eg, 1:10 diluted bleach, glutaraldehyde, food grade ethyl alcohol, chlorhexidine based products); thermal eradication; copper-silver ionization; Chlorine dioxide; ultraviolet light; ozone; disinfectant combinations (eg, Adec® ICX Adex, Newburg, OR)); sodium percarbonate; silver nitrate and cationic surfactant and silver ion catalyst.

  Prevent and / or control (ie slow down, delay, inhibit) biofilm development, disrupt biofilm, or reduce the amount of biofilm on the inside or outside surface of a water pipe or water delivery vehicle. Alternative antimicrobial agents that can be used to reduce include a particulate BT compound (or a composition comprising at least one particulate BT compound). The particulate BT compound can be introduced manually or automatically into water, water, and water delivery vehicles as a gel, spray, paste, liquid or powder, or other form known to those skilled in the art. In certain embodiments, the particulate BT compound, in either powder or liquid form, is mixed with at least one additional component, which comprises at least one additional biologically active component and / or biologically inactive component. The product is formulated with the active excipient and delivered or injected periodically into a water line, water delivery vehicle or water system. Compositions can be prepared by one of skill in the art using any number of methods known in the art. By way of example, an antimicrobial effective amount of a particulate BT compound can be used in combination with DMSO. In conventional use, a level of particulate BT compound that is sufficient to prevent biofilm formation is desired. However, in other embodiments, the level of the particulate BT compound is higher to reduce, remove, disrupt, or eliminate biofilms present in water lines, water delivery vehicles or water pipe systems. There is.

  The particulate BT compound can also be formulated to release slowly from compositions containing the particulate BT compound applied to a water mains, water delivery vehicle or water pipe system. The particulate BT compound can also be incorporated into the coating, which can be applied to the inner surface of a water pipe, vehicle or system, fixed, glued, or in some way arranged to contact. . The composition comprising the particulate BT compound can be a gel (eg, a hydrogel, thiomer, airgel or organogel) or a liquid. The organogel may include an organic solvent, lipoic acid, vegetable oil or mineral oil. The slow release composition may provide an antimicrobial 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.

  The particulate BT compound (or composition comprising the particulate BT compound), when administered in combination, has at least one or more other antimicrobial or enhanced synergistic antimicrobial effects as described herein. It can be combined with an antimicrobial agent (ie, a second, third, fourth, etc. antimicrobial agent). By way of example, enhanced antimicrobial activity may be observed when the particulate BT compound is administered together with an iron chelating antimicrobial agent. The particulate BT compound may be combined with at least one of an oxidizing agent, a microbicide or a disinfectant. A particulate BT compound prepared with a hydrophobic thiol (eg, thiochlorophenol) is used, which has a greater ability to adhere to water pipes and the surfaces of water delivery vehicles and systems than does a low hydrophobic BT compound. Can be shown. BT compounds with 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. Because of the chemical and physical properties of baking soda, it has a wide range of uses such as cleaning, 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 mixture of powders or dissolved or suspended in a powder, spray, gel, paste or liquid described herein. In another embodiment, the particulate BT compound is useful for maintaining a desired alkaline pH and with other alkali metal bicarbonate or carbonate materials (eg, potassium bicarbonate or calcium carbonate) that also possess purifying and deodorizing properties. Can be combined.

  As an additional example, the particulate BT compound (or composition comprising the particulate BT compound) can be combined with one or more of the following. Antimicrobial agents: for example, 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 compound); alkyl hydroxybenzoate; cationic antimicrobial peptide; aminoglycoside; quinolone; lincosamide; penicillin: cephalosporin; Tetracycline; other antibiotics known in the art; Coleus forskoli essential oil; silver or silver colloid antimicrobial agents; tin or copper based antimicrobial agents; Le; oregano; time; rosemary; or other herbal extracts; and grapefruit seed extract. Anti-caries agents: for example, sodium 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. Biomolecules: for example, bacteriocins. Preservative. Opacifier. pH adjuster. Sweetener. Surfactants: for example, anionic, nonionic, cationic and zwitterionic or amphoteric surfactants, saponins from plant material (see, for example, US Pat. No. 6,485,711). Particle abrasives: for example, silica, alumina, calcium carbonate, dicalcium phosphate, calcium pyrophosphate, hydroxyapatite, trimetaphosphate, insoluble hexametaphosphate, aggregated particle abrasive, chalk, finely pulverized natural chalk, and the like. Humectants: 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 such as polyacrylate and carboxyvinyl polymers such as Carbopol®. Polymeric compounds that enhance delivery of active ingredients such as antimicrobial agents. Buffers and salts to buffer the pH and ionic strength of the oral care composition. Bleach: for example, a peroxy compound (for example, potassium peroxydiphosphate). Foaming system: for example, sodium bicarbonate / citric acid system.

  In another embodiment, the particulate BT compound (or composition comprising the particulate BT compound) described herein controls biofilm development, disrupts biofilm, or reduces the amount of biofilm. For this purpose, 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 the LuxS-dependent pathway or interspecies quorum sensing signal (see, eg, US Patent 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, for example, 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 locally for disintegration and inhibition of bacterial biofilms and for treatment of periodontal disease (eg, See, for example, U.S. Patent No. 6,726,898).

  The effectiveness of the particulate BT compound as an anti-biofilm agent is to heat the water pipe, water delivery vehicle or water pipe system by heating the water pipe, water delivery vehicle or water pipe system to which the particulate BT compound is applied. Can be enhanced. In certain embodiments, the water pipe, water delivery vehicle or water pipe system is heated to about 37C to about 60C or about 37C to about 100C. In other embodiments, the water pipe, water delivery vehicle or water system is from about 45C to about 50C; from about 50C to about 55C; from about 55C to about 60C; from about 60C to about 70C; Heated to about 70C to about 80C; about 80C to about 90C; or about 90C to about 100C. 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 pipe system is heated will depend on the temperature applied. For example, the length of time required to achieve the same antimicrobial action is such that the water pipe, water delivery vehicle or water pipe system is heated to a lower temperature than would be required if heated to a higher temperature. The case is longer. Determination of the appropriate length of time for exposure of the water line, water delivery vehicle, or water system at each temperature can be readily determined by one skilled in the art.

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

  In another embodiment, a composition comprising particulate bismuth-thiol and use for tooth restoration, comprising a particulate BT compound, and dental amalgam and particulate BT compound and dental for use in the prevention and / or treatment of caries A composite material is provided herein. Generally, the only treatment for caries lesions is restoration of the tooth 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 restoration failure, especially when dental composites are used for restoration. The presence of bacteria that inhabit the interface between the composite and the tooth tissue may be an important factor in repair failure (see, for example, 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 mid-study. Recurrent peripheral caries accounted for 66% (32/49) and 88% (113/129) failure rates, respectively, for the main reasons for failure in amalgam and composite repair (Bernardo et al. JADA 2007; 138: 775-83). Polymerization shrinkage, a shrinkage that occurs during the composite curing process, is closely tied to the primary reason for postoperative peripheral leakage (eg, Estefan et al., Gen. Dent. 2003; 51: 506-509). reference).

  Attempts to incorporate antimicrobial compounds and agents into restorative materials such as dentin-binding systems (DBS) have been attempted with limited success. The development of composites and amalgams and other restorative materials with antimicrobial properties may 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 compositions described herein in the art with the particulate BT compounds described in the present invention, Provide antimicrobial activity, solubility and bioavailability, anti-biofilm effects, non-toxicity, enhanced antibiotic efficacy, and other properties as described herein. Intended to be.

  In certain embodiments, a composition is provided that includes a particulate BT compound and a dental composite. 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); Powers et al. , Dental Materials: Properties and Manipulation (New York: Mosby) (2007); Roeters et al. , J. et al. Dent. 32: 371-77 (1998)).

In another embodiment, there is provided a composition comprising a particulate BT compound and amalgam. 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, thus silver is provided as an alloy with other elements. This alloy is often referred to as a dental amalgam alloy, or collectively, the alloy is known as an "alloy for dental amalgam" (e.g., International Standards Organization Standardization ISO 1559, Dental Materials-Alloys fors). Dental Amalgam (1995)). Several types of dental amalgam alloys are known, all containing tin, most with some copper, and to a small extent zinc. Some of the dental amalgam alloys themselves contain small amounts 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. So-called dispersed amalgam alloys have about 70% silver, 16% tin and 13% copper. Different groups of amalgam alloys can 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. Thus, the mercury content of the finished dental amalgam restoration is about 50% by weight. In conventional dental amalgam alloys, the ratio of silver to tin results in a crystal structure that is essentially an intermetallic Ag3Sn called the gamma (γ) phase. The exact percentage of this phase controls the kinetics of the amalgamation reaction, as well as numerous properties of the resulting amalgam structure. In high copper dispersion alloys, the microstructure is usually a mixture of a gamma phase and a eutectic silver-copper phase. Different manufacturers have different amalgam alloy formats, but they are usually spherical or irregular, and are available as fine particles with a particle size of about 25-35 microns (Scientific Committee on Emerging and Newly Identified Health). Risks (SCENHR), European Commission: Directrate-General, Health & Consumer Protection, May 6, 2008 at Internet site:
ec. europa. eu / health / ph_risk / commites / 04_scenihr / docs / scenihr_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 the occurrence or recurrence of caries and / or inflammation, respectively) To reduce). The composition comprising the particulate BT compound can be a mucoadhesive composition applied to the surface of the teeth and / or gingiva or oral mucosa, adhere to the surface to some extent, or provide 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 slowly from the composition applied to the teeth. For example, the composite can be a gel (eg, a hydrogel, thiomer, airgel 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 can be applied inside or outside amalgam or composites or other restorative compositions. The slow release composition comprises administering 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 of ordinary skill in the art using any number of methods known in the art.

  Compositions comprising particulate BT compounds that are useful for tooth restoration include glass ionomer cements; giomers (formed by reacting fluoride-containing glasses and liquid polyacids); compomers (polymerizable dimethacrylate resins and ions). (Filterable glass-filled particles). The compomer may further include fluoride.

  Compositions comprising a particulate BT compound applied to the surface of a tooth, amalgam or composite may further include one or more other surfactants that enhance antimicrobial activity. Exemplary antimicrobial agents for use in compositions containing the particulate BT compound include, for example, chlorhexidine, sanguinarine extract, metronidazole, quaternary ammonium compounds (eg, cetylpyridinium chloride); bisguanides (eg, chlorhexidine diglucose). Nato, hexetidine, octenidine, alexidine); and halogenated bisphenolic 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 Essential oils of ICX, Coleus forskoli; silver or silver colloid antimicrobial agents; tin or copper based antimicrobial agents; chlorine or bromine oxidizers, manuka oil; oregano; thyme; rosemary; 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 and the like.

  The composition may further comprise 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 anionic, non-ionic, cationic and zwitterionic or amphoteric surfactants, or may include saponins from plant material (see, eg, US Pat. 6,485,711). Buffers and salts to buffer the pH and ionic strength of the compositions for oral use may also be included. Other optional ingredients that may be included are bleaching agents, such as peroxy compounds; potassium peroxydiphosphate; effervescent systems, such as sodium bicarbonate / citric acid systems.

Compositions comprising particulate bismuth-thiol and uses for oral hygiene and for treating oral inflammation and infection In another embodiment, compositions comprising particulate BT compounds are formulated for oral use, May be used in methods for preventing or reducing microbial growth of and for preventing and / or treating oral microbial infection and inflammation. Therefore, these compositions are useful for preventing or treating plaque, bad breath, periodontal disease, gingivitis, and other infections of the mouth (i.e., reduce or inhibit their onset, and Useful to reduce likelihood). An oral composition comprising a particulate BT compound can prevent and / or control (ie, slow down, delay, inhibit) biofilm development, disrupt biofilm, or disturb oral surfaces, particularly teeth or It can also be useful in reducing the amount of biofilm present on the gums.

  Entrapped food particles, poor oral hygiene and poor oral health, and improper cleaning of dentures can promote microbial growth on teeth, peri-gingival, and on the tongue. The continued development of microorganisms and the presence of caries can result in halitosis, plaque (ie, biofilm formed by microbial colonization), gingivitis, and inflammation. Without adequate oral care (eg, brushing, flossing), more severe infections, such as periodontal disease and jaw infections, can result.

  Good oral hygiene is important not only for oral health but also for the prevention of several chronic conditions. Controlling bacterial growth in the mouth 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 are at increased risk of developing severe gum problems, and lowering 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, low birth weight infants.

  Bacteria are major etiological agents 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, the gingival epithelium, and the oral environment, which makes it difficult to treat periodontitis. Fungicides and antibiotics currently used to treat such infections often do not kill all of the problematic organisms. The use of drugs that are ineffective against certain strains can result in the growth of resistant strains. Moreover, these drugs can induce undesired side effects, such allergic reactions, inflammation, and discoloration of the teeth.

  Bacterial dental plaque is a biofilm that adheres stubbornly to tooth surfaces, restorations, and prostheses. The primary means of controlling the biofilm in the mouth is by mechanical cleaning (ie, brushing, flossing, etc.). If not subjected to such washing, within the next two days, the tooth surface will be predominantly stained by facultative Gram-positive cocci, mainly of the genus Streptococcus. Bacteria secrete an extracellular mucus layer that helps fix the bacteria to the surface and protects the attached bacteria. When the tooth surface is covered with attached bacteria, microcolonization begins. Biofilms grow primarily through cell division of adherent bacteria, rather than the attachment of new bacteria. The doubling time of plaque formed by bacteria is rapid during early development and slower in more mature biofilms.

  If the engraftment continues to adhere to bacteria already attached to the pellicle, coaggregation occurs. Co-aggregation results in the formation of a complex array of different bacteria that are interconnected. A few days after uninterrupted plaque forms, the gingival margin becomes inflamed and swells. Inflammation can form deep gingival sulcus. The biofilm extends to this subgingival area and grows in this protected environment, forming a mature subgingival plaque biofilm. No gingival inflammation is observed until the biofilm changes from being largely composed of Gram-positive bacteria to one containing Gram-negative anaerobic bacteria. Subgingival bacterial microcolonies, mainly composed of gram-negative anaerobic bacteria, become established in the gingival sulcus between 3 and 12 weeks after the onset of supragingival plaque formation. 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 defense. For example, antibiotic doses that kill airborne bacteria need to be increased by a factor of 1500 over doses that kill biofilm bacteria. At this high concentration, these antimicrobial agents also tend to be toxic to patients (see, for example, Coghlan 1996, New Scientist 2045: 32-6; Elder et al., 1995, Eye 9: 102-9). .

  Careful and frequent physical removal of bacterial plaque biofilms 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 dental hygienists or dentists is an essential component in the prevention and treatment of periodontitis.

  In certain embodiments, the particulate BT compound is an oral hygiene composition that can be used routinely by any subject, such as toothpaste, mouthwash (ie, mouth rinse), oral gel, toothpaste, oral spray (oral inhalation) Such as sprays dispersed by a container), edible films, chewing gums, oral slurries, denture cleansers, denture preservatives, dental floss, and the like, for example, on a coating or in a device. 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 flossing and cleaning tools, and equipment. It 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 compound described herein, Disclosed herein, including antimicrobial activity, solubility and bioavailability ranges, anti-biofilm effects, non-toxicity, enhanced antibiotic potency, and other properties described herein It is intended to provide advantages.

  The particulate BT compound can be used to prevent or treat dental caries and / or inflammation (ie, to reduce the likelihood of the occurrence or recurrence of dental caries and / or inflammation, respectively, by administering the particulate BT compound to the tooth surface. ) 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 for slow release 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 contain an organic solvent, lipoic acid, vegetable oil, or mineral oil. Such a gel or liquid coating formulation may be applied to the interior or exterior of the 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 for weeks, or for 1, 2, 3, 4, 5, or 6 months. Such compositions can be prepared by one of ordinary skill in the art using any number of methods known in the art.

  In certain other embodiments, an antimicrobial composition comprising a particulate BT compound and one or more additional antimicrobial compounds or agents, as described herein, is provided for oral use. Particularly useful are compositions comprising a second antimicrobial agent that, when administered in combination, has an enhancing or synergistic antimicrobial effect as described herein. By way of example, an enhanced antimicrobial effect may be observed when the particulate BT compound is administered with an iron chelating antimicrobial agent. In certain other 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 a greater ability to adhere to tooth and mouth tissue than a less hydrophobic BT compound. May be shown large. BT compounds with 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 include baking soda or other alkaline compounds or substances. 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. The 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 fine particle BT compound may further contain one or more of the following components.
Antimicrobial agents: for example, chlorhexidine: sanguinarine extract, metronidazole, quaternary ammonium compounds (for example, cetylpyridinium chloride); bisguanides (for example, chlorhexidine digluconate, hexetidine, octenidine, alexidine); halogenated bisphenol compounds (for example, 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; essential oils of Coleus forskoli; silver or silver colloid antimicrobial agents; tin or copper based antimicrobial agents; 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, omega-3 fatty acids, gamma-linolenic acid (GLA), green tea , Ginger, grape seed etc. Anti-caries agents: for example, sodium 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. Desensitizers: 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. Biomolecules: for example, bacteriocins, bacteriophages, antibodies, enzymes and the like. Flavoring agents: for example, peppermint and spearmint oils, fennel, cinnamon and the like. Proteinaceous material: for example, collagen. Preservative. Opacifier. Coloring agent. pH adjuster. Sweetener. Pharmaceutically acceptable carriers: for example, starch, sucrose, water or water / alcohol systems and the like. Surfactants: for example, anionic, nonionic, cationic and zwitterionic or amphoteric surfactants, saponins from plant material (see, for example, US Pat. No. 6,485,711). Particle abrasives: for example, silica, alumina, calcium carbonate, dicalcium phosphate, calcium pyrophosphate, hydroxyapatite, trimetaphosphate, insoluble hexametaphosphate, aggregated particle abrasive, chalk, finely pulverized natural chalk, and the like. Humectants: 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 such as polyacrylate and carboxyvinyl polymers such as Carbopol®. Polymeric compounds that enhance 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, a peroxy compound (for example, potassium peroxydiphosphate). Foaming system: for example, sodium bicarbonate / citric acid system. Discoloration system. In certain embodiments, the abrasive is silica or finely divided natural chalk.

  The oral hygiene composition comprising the particulate BT compound formulated for use as a toothpaste further comprises a moisture absorbent (eg, glycerol or sorbitol), a surfactant, a binder, and / or a flavoring agent. Is also good. Toothpastes may contain sweetening, whitening, preservative, and antimicrobial agents. The pH of compositions for toothpastes 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 agents, monoolein Acid sorbitan, saccharin sodium, tetrasodium pyrophosphate, methylparaben, propylparaben. If desired, one or more colorants can be used, for example, FD & C Blue. Other suitable components that can be included in a toothpaste formulation are described in the art, for example, in US Pat. No. 5,560,517.

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

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

  In yet another specific embodiment, the oral hygiene compositions described herein comprise at least one of the following for controlling biofilm development, disrupting biofilms, or reducing the amount of biofilms: Or more anti-biofilm agents. As understood in the art, interspecies quorum sensing involves biofilm formation. Certain agents that increase the LuxS-dependent pathway or interspecies quorum sensing signal (see, eg, US Patent No. 7,427,408) contribute to the control of biofilm development and / or growth. Exemplary agents include, for example, N- (3-oxododecanoyl) -L-homoserine lactone (OdDHL) blocking compounds and N-butyryl-L-homoserine lactone (BHL) analogs, either in combination or separately. (See, for example, 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 locally for the disintegration and inhibition of bacterial biofilms and for the treatment of periodontal disease (eg, for example, in the United States). Patent No. 6,726,898).

  The oral hygiene compositions described herein may contain a sufficient amount of the particulate BT compound to perform a substantial antimicrobial action at the time required for normal brushing, 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 completion of 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 mucoadhesive polymers or other agents that contribute to the retention of the particulate BT compound on mucosal, tooth and restoration surfaces. The particulate BT compound may be added to a stable, viscous, mucoadhesive aqueous composition, which is used to prevent and treat mucosal ulcerative, inflammatory, and / or erosive disorders, and / or for topical treatment. It may be used for delivery of a pharmaceutically active compound to mucosal surfaces or transfer into the systemic circulation (see, for example, US Pat. No. 7,547,433).

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

  In another embodiment, the oral hygiene composition comprising the particulate BT compound may further comprise a mucosal germicidal preparation for topical application in the mouth. The oral hygiene composition may further include an aqueous slurry useful for cleaning 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 (cavities). It may further contain one kind of mint. One such mint, referred to as CaviStat® (Ortek Therapeutics, Inc., Roslyn Heights, NY), contains arginine and calcium, helps neutralize acidic pH, and deposits calcium on enamel surfaces. To promote. The 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.

  In another embodiment, an adhesive composition comprising particulate bismuth-thiol formulated for dental and orthopedic use, wherein the composition comprising particulate BT compound is on a bone or joint replacement (prosthesis), or It is formulated for use in a method for preventing or reducing microbial growth of tissues and skeletal structures adjacent to bone or joint replacements (artificial joints). In certain embodiments, 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 treating is provided. In some embodiments, the composition comprises a particulate BT compound and a bone cement, and in some other embodiments, a particulate BT compound and a dental cement. Accordingly, these compositions prevent and / or treat (ie, reduce or inhibit the onset of) microbial infections of the skeleton and supporting structures (ie, bones, joints, muscles, ligaments, tendons), such as osteomyelitis. The likelihood of its occurrence or recurrence). The compositions described herein comprising a particulate BT compound and bone cement or dental cement prevent and / or control (ie, slow down, delay, inhibit) biofilm development, disrupt biofilms Also useful for reducing or reducing the amount of biofilm present in or on a joint, bone, ligament, tendon or tooth, or replacement joint, bone (partially or entirely), ligament, tendon or tooth surface It can be.

  Cements described herein and known in the art are binder materials that can bind materials together and cure. Such materials may bind the tissue together or may connect a prosthetic or prosthetic device (eg, a prosthetic joint, bone or tooth) to adjacent tissue. Bone cements include, for example, polymethyl methacrylate (PMMA), magnesium phosphate and calcium phosphate. The form of calcium phosphate is used as a "replacement bone" to treat bone cracks and cuts that cannot heal sufficiently quickly and / or properly without graft material. Compositions comprising bone cement (eg, calcium phosphate) and a particulate BT compound 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 implantation site.

  In certain embodiments, the compositions described herein that are useful as bone cements include a BT compound or a particulate BT compound, and a preparation of calcium 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 or calcium phosphate bone cement, or magnesium phosphate-containing 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 head of print) cement. Magnesium phosphate used in the art is also referred to as struvite (MgNH4PO4 * 6H2O) cement (see, for example, Grosshardt et al., Tissue Eng. Part A, 2010 Jul 30, e-pub ahead of print; see, e.g., Bohner et al.). al., J. Pharm. Sci. 86: 565-72; (1997); Fuller et al., 3: 299-305 (1992); Lovenhoffer et al., J. Orthopedic 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 head of print. In certain other embodiments, the composition comprising the particulate BT compound and calcium 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 It may further include one or more anti-inflammatory agents.

  Bone cement has been used in the art for the release of drugs and active substances. In certain embodiments, the calcium phosphate cement can be in the form of hydroxyapatite microspheres, at least in part, encapsulating an agent (eg, an antimicrobial agent) for therapeutic use (eg, US Pat. 730, 324). Such cements containing microspheres are useful for slow release of active agents contained within the microspheres. Contemplated herein are compositions comprising calcium phosphate microspheres comprising a particulate BT compound.

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

  Also provided herein is a composition comprising a particulate BT compound and a dental cement (ie, a dental adhesive), wherein the composition is for inhibiting, preventing or treating microbial infection of a tooth or gingiva. 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. 071,528); bismuth oxide (for example, see Bueno et al., Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endod. 107: e65-69 (2009)); MTA) (see, for example, Hwang et al., Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endod. 107: e96-102 (2009)).

Embodiments of the present invention disclose herein an antimicrobial agent formulated using dental cement or bone cement described in the art, replacing the particulate BT compound according to the present invention. Providing advantages such as antimicrobial activity, solubility and bioavailability, anti-biofilm effects, non-toxicity, enhanced antibiotic efficacy, and other properties as described herein. Intended. Bone and dental cements can be formulated with particulate BT compounds and one or more additional components according to methods described in the art (eg, US Patent Application Publication No. 2006/0205838; Alt et al. 48: 4-84-88 (2004); Bohner et al., Supra; Bueno et al., Supra; 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 including bone cement or dental cement can range from about 10-500 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 antibiotics currently used in bone and dental cements. . Compositions described herein that include a particulate BT compound and bone cement (eg, calcium phosphate) or dental cement may further include one or more additional antimicrobial compounds or agents. Particularly useful are compositions comprising a particulate BT compound, and a second antimicrobial agent that, when combined, has an enhanced or synergistic antimicrobial effect as described herein. As an additional example, enhanced antimicrobial activity may be observed when the particulate BT compound is administered with an iron chelating antimicrobial agent. In certain other embodiments, the particulate BT compound is formulated with an anti-inflammatory agent, compound, small molecule or macromolecule (eg, a peptide or polypeptide).

  Compositions comprising the particulate BT compound and bone cement as described herein can be used to attach, stabilize, or fix fractures, fusions, osteotomies, or replacement joints (e.g., metal products). , Screws, plates, staples, pins and wires, etc.). A composition comprising a particulate BT compound and a dental cement as described herein may be used to form a dental pulp, tooth cap, inner layer, tooth, or a dental filling or restorative composition in a tooth. Can be used for coating. These compositions can be applied to adhere and adhere to the surface of bone and / or joint-related metal products, or can be formulated into coatings that can be placed in contact in some way. In certain embodiments, the coating comprises a particulate BT compound and a calcium or magnesium phosphate bone cement. The particulate BT compound and calcium or magnesium phosphate are formulated together for application to a bone metal product according to methods practiced in the art. For example, a composition comprising a particulate BT compound and a bone cement (eg, a calcium or magnesium phosphate bone cement) can be a liquid, gel, paste or spray (eg, a thermal spray including a plasma spray) for application to metal products. It can be in the form. The composition comprising the particulate BT compound and the bone cement can be a gel (eg, a hydrogel, thiomer, airgel or organogel) or liquid. The organogel may include an organic solvent, lipoic acid, vegetable oil or mineral oil. The slow release composition may provide an antimicrobial 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. 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 has at least one other antimicrobial agent that has an enhanced or synergistic antimicrobial effect (ie, greater than an additive effect) when administered in combination (Ie, second, third, fourth, etc. antimicrobial agents). By way of example, enhanced antimicrobial activity may be observed when the particulate BT compound is administered together with an iron chelating antimicrobial agent. In certain embodiments, a composition comprising a particulate BT compound and bone cement or dental cement may be combined with at least one other antimicrobial and / or anti-inflammatory agent selected from: Chlorhexidine; sanginalin 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 antimicrobial peptides; aminoglycosides; quinolones; lincosamides; Sporin; macrolide; tetracycline; other antibiotics known in the art; Coleus forskoli essential oil; silver or silver colloid antimicrobial agents; tin or copper based antimicrobial agents; Marie; 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, omega-3 fatty acids, gamma-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 comprises 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 may optionally include 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 with dental cements and bone cements as described herein and in the art (eg, Akashi et al., Biomaterials 22: 2713-17 (2001); US Patents). No. 6,071,528; Alt et al., See above).

  Animal models of xenobiotic infection can be used to characterize the antimicrobial activity of a composition comprising a particulate BT compound and death song cement 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 antimicrobial agents to kill stationary phase microorganisms as well as those that are adherent to foreign substances (eg, Widmer et al. J. Infect. Dis. 1990; 162: 96-102; Widmer et al. Antimicrob Agents Chemother 1991; 35: 741-6; see also, for example, Karchmer. Clin. Infect. Dis. 1998; 27: 714-6).

  By way of non-limiting example, and for illustrative purposes only, bone cement comprises 75% (2/2) methyl methacrylate styrene copolymer, 15% polymethyl methacrylate (to assist in handling the composition) and 10% barium sulfate ( (For radiopacity), as well as in about 10 to about 500 μg of the particulate BT compound / g of cement powder (ie, 0.001-0.05% w / w). 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 formation (ie, slow, delay, Suppress), disintegrate the biofilm, or reduce the amount of biofilm present on the painted surface by applying the particulate BT compound described herein in the paint or as a paint coating on the paint. Intended to be incorporated. The compositions described herein comprising the particulate BT compound can be used in a number of products, such as medical devices, orthopedic devices, dental devices, industrial devices, electronic devices, walls, floors, ceilings, roofs. Piles, piers, piers, conduits, pipelines and piping structures (eg, intake screens, cooling towers), heat exchangers, dams and fabrics, and other surfaces, eg, cars, trains, airplanes and ships, eg, large It may be formulated with a paint or paint coating applied to any one of the surfaces present in and on, but not limited to, 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 biofouling or biofilms formed on products exposed to water. is there. The formation of biofilms on surfaces in marine environments is thought to be an important contributor to the colonization and recruitment of some fixed invertebrate communities on marine structures (eg, Siboni et al. , FEMS Microbilol Lett 2007; 274: 24-9). Subsequent interaction of the macrobiota with these microbial membranes will result in the invertebrates and algae (which are responsible for most of the hydraulic drag associated with sexual attachments) within days or weeks. (See, e.g., Schultz, Biofouling 2007; 23: 331-41). Old biofilms on the surface supported barnacle larval attachment, regardless of the type of support (see, for example, Hunga et al., J Exptl Marine Biol Ecol 2008; 361: 36-41). Microfilaments are also present in one or more significant developmental stages at one or more significant stages in the ascidians Phallasia nigra, the polychaete mosquito Kanesan Kanji (Hydroides elegans), and the barnacles Balticus barnacle (Balanus amphirite). (See, e.g., Zardus et al., Biol Bull 2008; 214: 91-8). Biofilms can also enhance the adhesion of the mussel, the mussel, Dressena polymorpha, to some artificial surfaces (see, eg, Kavouras & Maki. Inverteb Biol 2005; 122: 138-51). Hundreds, if not billions, of billions of dollars in gains and losses for marine food, power generation and manufacturing, and for water and wastewater treatment facilities, have created significant damage to the ecosystem in which mussels are introduced. Is causing.

  In seawater, brackish and freshwater environments, organisms assemble, inhabit, attach and proliferate on underwater structures and ships. Such organisms include algae, fungi and other microorganisms, as well as aquatic animals such as ascidians, hydroids, bivalves, bryozoans, polychaetes, sponges, barnacles. The presence of these organisms, known as "fouling" of the structure, for example, adds to the weight of the structure and / or interferes with its hydrodynamics, thereby reducing its operating efficiency, corroding Increased ease and destruction or crushing of the structure can be harmful.

  Certain paints and coatings that have been used to prevent or reduce biofouling and biofilm production have been shown to inhibit biofouling and biofilm formation while maintaining desirable and beneficial flora and Contains toxic components that can be toxic to the fauna. Exemplary biocides and chemical toxins include copper and copper-containing compounds (eg, cuprous oxide), mercury, arsenic, tributyltin oxide (TBT), organotin (ie, having one or more carbon groups attached thereto). Tin), hexio 2 parts 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 provide the benefits disclosed herein, such as antimicrobial activity, a range of solubility and bioavailability, anti-biofilm effects Provides enhanced antibiotic efficacy, as well as other properties as described herein. The particulate BT compound is substituted for other antimicrobial agents in paints and paint coatings and is described herein using methods known to produce paints and paint coatings that include biocides. The particulate BT compounds and methods described can be incorporated into these paints and paint coatings by integration of the methods (eg, US Pat. Nos. 4,596,724; 4,410,642; 4,788,302). 5,470,586; 6,162,487; 5,384,176; U.S. Patent Application Publication Nos. 2007/125703 and 2009/0197003; Gerhart et al., J. Chem. Ecol. 14: 1905-17 (1988); Sears et al., J. Chem. Ec. 16: 791-99 (1990); Ganguli et al., Smart Mater. Struct. 18: 104027 (2009); Cao et al., ACS Applied Materials Interfaces 1: 494 (2009); Materials 7: 236-41 (2008)). Examples of the paint into which the fine particle BT compound can be mixed include a paint based on epoxy, silicone or acrylic. In more particular embodiments, the particulate BT compound can be incorporated into paints formulated for marine use and exposure to seawater, such as alkyd resin-based, bitumen-based, Gilsonite-based Base material paints, chlorinated rubber base materials and epoxy resin base material paints can be mentioned.

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

  The particulate BT compound can also be formulated to release slowly from a composition comprising the particulate BT compound applied to a painted surface. The particulate BT compound can also be incorporated into a coating (eg, an epoxy coating) that can be applied to adhere and adhere to the surface of the painted structure or product, or placed in contact in some manner. Particulate BT compounds can be slowly released from such compositions. Slow release compositions comprising a particulate BT compound can be gels (eg, hydrogels, thiomers, airgels or organogels) or liquids. The organogel may include an organic solvent, lipoic acid, vegetable oil or mineral oil. The slow release composition may provide an antimicrobial 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.

  Other coatings used in the art and with which the particulate BT compounds described herein can be formulated include polysaccharides, such as polysaccharide matrices that are reversibly crosslinked with polyvalent metal cations (eg, Titanium nanotubes; nanostructured surfaces; biocompatible dextran-coated nanoceria (having pH-dependent antioxidant properties); polysorbone block polymers; and other biodegradable coatings (see US Patent Application No. 2009/0202610). For example, see also US Pat. No. 6,162,487). Other coatings contemplated herein formulate particulate BT compounds with anti-corrosion and anti-fouling anti-microbial coatings used in the industry, including, but not limited to, carnauba wax fluoropolymer, xylan (registered trademark). 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 into a paint or paint coating has an enhanced or synergistic antimicrobial effect as described herein when administered in combination. , At least one other antimicrobial agent (ie, a second, third, fourth, etc. antimicrobial agent). By way of non-limiting example, antimicrobial agents that may be included in the composition containing the particulate BT compound include chlorhexidine; sanguinarine extract; metronidazole; quaternary ammonium compounds (eg, cetylpyridinium chloride); bisguanides (eg, chlorhexidine Gluconate, hexetidine, octenidine, alexidine); halogenated bisphenolic compounds (eg, 2,2'-methylenebis- (4-chloro-6-bromophenol) or other phenolic antibacterial compounds; alkylhydroxybenzoates; cationic Aminoglycosides; quinolones; lincosamides; penicillins: cephalosporins; macrolides; tetracyclines; other antibiotics known in the art; essential oils of Coleus forskoli; Manuka oil; Oregano; Thyme; Rosemary; or other herbal extracts; and Grapefruit seed extract. Surfactants, diluents or carriers, buffers and / or bleaches, as described above and herein.

  Compositions Comprising Fine Particulate Bismuth-Thiol Formulated with Concrete and Cement Compounds Certain other embodiments are biofilms that prevent and / or control (i.e., slow, delay, inhibit) biofilm development. In order to disintegrate or reduce the amount of biofilm present on the concrete surface, in industrial cement, and in or on concrete, mortar and grout (including concrete, mortar and grout coatings) It is intended to incorporate the particulate BT compounds described in this document. Microorganisms growing on and in concrete structures reduce the useful life of the product and can have health hazards to animals and humans exposed to microorganisms present on concrete surfaces (see, for example, Idachaba et al., 19: 284-91 (2001); Idachaba et al., J. Hazard. Mater. 90: 279-95 (2002);

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

  The compositions described herein, including particulate BT compounds, may be used in concrete structures such as bridges, buildings, pipes, elevated highways, tunnels, garages, offshore oil pits, wharfs, dam walls, water systems and pipelines. Can be used or coated with cement used for floors, countertops, sidewalks, driveways, loading / unloading piers, skateboard yard structures, and structures for primary storage of radioactive waste obtain. The particulate BT compounds described herein can be incorporated into cements 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, for example, sulfur oxidizing bacteria (Thiobacillus thiooxidans). As a non-limiting example by way of example, the bismuth thiol compound BisEDT (but not the particulate BT compound described herein) slows 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 hindered concrete strength. Other compounds, such as BisPYR, may be useful for controlling fouling and biofilm development by molds and algae. Embodiments of the present invention are directed to replacing the bismuth-thiol compounds and other antimicrobial agents with the particulate BT compounds described in the present invention to provide the advantages disclosed herein, such as antimicrobial activity, solubility, and biological properties. It is intended to provide bioavailability, anti-biofilm activity, non-toxicity, enhanced antibiotic efficacy, as well as other properties as described herein.

  The particulate BT compound can be manually or automatically introduced to the concrete surface as a gel, spray, paste, liquid or powder or other form known to those skilled in the art. In certain embodiments, the particulate BT compound, in either powder or liquid form, is mixed with at least one additional ingredient, which comprises at least one additional biologically active ingredient and / or biologically inactive ingredient. Formulate the product with the active excipient and deliver it regularly in or on the concrete structure (ie, to the surface of the exposed concrete structure, especially to the surface exposed to water) Or be injected. Compositions can be prepared by one of skill in the art using any number of methods known in the art. As an example, an antimicrobial effective amount of a particulate BT compound can be used in combination with DMSO (eg, 1 mg / ml particulate BT compound in DMSO). In conventional use, a level of particulate BT compound that is sufficient to prevent biofilm formation is desired. However, in other embodiments, the level of the particulate BT compound may be higher to reduce, remove, disintegrate, or eliminate the biofilm present on the concrete surface.

  The particulate BT compound may also be formulated to release slowly from a composition comprising the particulate BT compound applied to the surface of the concrete structure. The particulate BT compound can also be incorporated into a coating (eg, an epoxy coating), which is applied to the surface of the concrete structure and is fixed, glued, or otherwise arranged to contact. Can be done. Particulate BT compounds can be slowly released from such compositions. Slow release compositions comprising a particulate BT compound can be gels (eg, hydrogels, thiomers, airgels or organogels) or liquids. The organogel may include an organic solvent, lipoic acid, vegetable oil or mineral oil. The slow release composition may provide an antimicrobial 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.

  The particulate BT compound (or a composition comprising the particulate BT compound), when administered in combination, has at least one or more other anti-microbial effects with enhanced or synergistic antimicrobial activity as described herein. It may be combined with an antimicrobial agent (ie, a second, third, fourth, etc. antimicrobial agent). By way of example, an enhanced or synergistic antimicrobial effect may be observed when the particulate BT compound is administered together with an iron chelating antimicrobial agent. The particulate BT compound can be combined with at least one other antimicrobial agent, including fungicides or algicides. By way of non-limiting example, antimicrobial agents that may be included in the composition containing the particulate BT compound include chlorhexidine; sanguinarine extract; metronidazole; quaternary ammonium compounds (eg, cetylpyridinium chloride); bisguanides (eg, chlorhexidine Gluconate, hexetidine, octenidine, alexidine); halogenated bisphenolic compounds (eg, 2,2'-methylenebis- (4-chloro-6-bromophenol) or other phenolic antibacterial compounds; alkylhydroxybenzoates; cationic Aminoglycosides; quinolones; lincosamides; penicillins: cephalosporins; macrolides; tetracyclines; other antibiotics known in the art; essential oils of Coleus forskoli; Manuka oil; Oregano; Thyme; Rosemary; or other herbal extracts; and Grapefruit seed extract. Surfactants, diluents or carriers, buffers and / or bleaches, as described above and herein.

  A particulate BT compound prepared using a hydrophobic thiol (eg, thiochlorophenol) is used, which has a greater ability to adhere to concrete surfaces, especially those exposed to water, than a low hydrophobic BT compound. Can be shown. BT compounds with 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 Some embodiments include the particulates described herein in or on engineered surfaces that include processed natural and synthetic rubbers and / or rubber coatings, such as silicone and silicone coatings. Incorporating BT compounds, such as 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, cars, tires, doors and window frames, fill pipes, belts, mats, dampers (anti-vibration cradle), trains, airplanes, ships, boats, submarines, piles, pipes, pipelines, pipes and fabrics, sounding / water Fittings, household goods, flooring, footwear, exercise equipment, mobile phones, computer equipment, Compounds with organic fillers, field products, such as roofing materials, awnings, tarpaulins, roof tarpaulins and swimming pool linings, and, for example, sterile products, and food and beverage preservatives, pharmaceuticals, and chemicals and It is intended to reduce biofilms and biofouling on such rubber surfaces in systems for water sterilization.

  The particulate BT compounds described in this invention can be used to integrate these and other natural and artificial BT compounds by integrating the BT compositions and methods described herein with processing steps known for these product classes. It can be incorporated into rubber products. As a non-limiting example by way of illustration, BT (but not the particulate BT described in the present invention) has been incorporated into hydrogel-coated polyurethane rods and Dacron grafts (Domenico et al. Antimicrob Agents Chemother 2001; 45: 1417-). Domenico et al., Peptides 2004; 25: 2047-53). WO / 2002/077095 and Japanese Patent Application No. 1997-34076 describe pre-vulcanized and / or vulcanized raw rubber formulations containing silver-based compounds to provide antimicrobial properties; U.S. Pat. Nos. 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 a rubber matrix. Drug-eluting silicone compositions can include an antimicrobial agent dispersed in a modified biologically active binder that can be applied to a medical device or other surface without using an inert polymeric carrier ( U.S. Patent Application 2009/0043388).

  Silicone oils generally have molecular weights in the range of 2,000 to 30,000 and viscosities 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 Stokes. Silicones are typically used in a variety of materials that suffer from microbial deposits. Examples of these include sealants, caulks, greases, oils, propellants, rubber, irrigation tubes and implants. Silicone-based antifouling coatings and other antimicrobial coatings have been described, but with poor efficacy, poor durability, poor biocompatibility, loss of antimicrobial activity, short useful life, and expensive Materials and other fabric-related drawbacks (e.g., Schultz J Fluids Eng 2004; 126: 1039-47; U.S. Patent No. 4,025,693; Yan & Li. Ophthalmologica 2008; 222: 245-8; U.S. Patent No. 6,221,498; U.S. Patent No. 7,381,751; European Patent Application EP0506113; Sawada et al. JPRAS 1990; 43: 78-82; Tiller et al. Surface Coatings Internationala. Part B: Coatings Transactions 2005; 88: 1-82; Juhni & Newby Procedures Annual Meeting Adhesion Society 2005; 28: 179-181; 19: 6167; U.S. Patent Publication No. 2009/0215924; Bayston et al. Biomaterials 2009; 30: 3167-73; Gottenbos et al.Biomaterials 2002; 23: 1417-23; Millsap en ae. ek 2001; 79: 337-43). These publications describe methods for incorporating antimicrobial materials into rubber products, but of the products or methods they describe, provided by the particulate BT described herein. Nothing offers any advantages.

  Accordingly, embodiments of the present invention are described herein in these and similar rubber (including silicone) products and methods, and in plastic and polymerized anti-methods, such as those referenced below. It is intended to replace the particulate BT. In each of these and other known processing situations, the particulate BT described herein may be incorporated according to the disclosure herein and provided by these particulate BTs instead of other antimicrobial agents. Such advantages may be provided herein, such as antimicrobial 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 fouling in or on silicone products. The particulate BT concentration (by weight) in the silicone can vary from as low as about 0.0001% to about 0.1%, for example, depending on the intended use and properties of the silicone rubber product. The particulate BT described herein may also be incorporated as a coating on silicone or in a silicone gel or oil to prevent or treat biofilms on silicone surfaces for extended periods of time. . Silicone rubber inlet valves are described in WO / 2008/064173, which periodically oozes out silicone oil and thus provides an effective antimicrobial level of the particulate BT described herein. The presence in natural exudates imparts anti-biofilm and / or anti-fouling capabilities to products containing such valves or similarly shaped silicone rubber devices. The erodible oil spreads on 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 on other surfaces exposed to water or humidity.

  For enhanced retention of BT on rubber surfaces, the particulate BT described herein may be used, for example, by using a hydrophobic thiol (eg, thiochlorophenol), which may have enhanced adhesion properties, and And / or including a 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, thereby providing greater hydrophobicity based on a particular thiol moiety. May be selected to retain sex. For example, the silicone material can be assembled at a temperature of 100 ° C. or less in the presence of a suitable concentration of the particulate BT described herein. The bioerodible material allows for the gradual release of such BT at levels that prevent biofilm formation, for example, at about 1-2 ppm. In other embodiments, rubber and / or plastic components that gradually elute the particulate BT compound and can be regularly replaced to prevent biofouling in various industrial systems or medical devices are contemplated. .

  In certain other embodiments, and similar to those described above with respect to compositions and methods for incorporation of BT 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 uses for such particulate BT-containing plastic articles 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, sounding / water fittings, homes in and on all types of vehicles, including cars, trains, planes and ships, boats, submarines, piles, pipes, pipelines and textiles Sundries, flooring, footwear, sports equipment, mobile phones, compounds with organic fillers, outdoor products such as roofing, sunshades, tarpaulins, roof waterproofing and swimming pool linings, and food and beverage preservatives Plastics in other products, including those used in pharmaceutical and chemical and water sterilization Include 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 demulsifiers, are often used, which serve as a nutrient source for microorganisms. Examples of such modern plasticizers include phthalate, adipate and other esters. These and other plasticizers are particularly susceptible to bacteria and fungi, especially in areas of high humidity, resulting in microbial surface growth and spore growth, which can lead to one or more infections, allergic reactions, in humans and animals, It can cause unpleasant odors, stains, embrittlement of plastics, premature product failure and other undesirable consequences.

  Modification of plastic products during or after processing steps by the introduction of antifouling and other antimicrobial coatings has been described, but typically has poor efficacy, poor durability, poor performance Suffers from poor biocompatibility, loss of antimicrobial activity, short useful life, expensive materials and other textile related drawbacks (eg, US Pat. Nos. 3,624,062; 4,086,297; No. 4,663,077; No. 3,755,224; No. 3,890,270; No. 6,495,613; No. 4,348,308; No. 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. dives 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 benefits provided by the particulate BT described herein. Nevertheless, it is generally known to those skilled in the art to incorporate an antimicrobial agent in or on a plastic product according to the following strategy: (a) Adsorption of the active substance on the polymer surface (B) introduction of the 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 attachment 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 the finished polymer.

  For example, the particulate BT described herein can be introduced into these and similar systems, either manually or automatically, as a gel, spray, liquid or powder. In one embodiment, the particulate BT, eg, in powder or liquid form, participates in components for plastic processing, eg, active components (eg, polymer precursors, catalysts, initiators, crosslinkers, etc.), as well as the production mixture. It is mixed with excipients (eg, carriers, solvents, release agents, dyes or colorants, plasticizers), which are periodically injected into the processing system. For example, a 1 mg / ml solution or suspension of particulate BT in DMSO is periodically injected into the polymer forming reaction or sprayed onto the working part of the molding unit to provide the desired anti-biologic in the finished product. Film density can be achieved.

  Thus, these and certain related embodiments disclosed herein may include one or more microparticle BTs, and optionally include a synergistic or potentiating antibiotic as described herein. The inclusion of the particulate BT compositions disclosed in the present invention in such products and processes, which may further include such antibiotics, is contemplated.

  According to certain embodiments described herein, non-limiting examples of bacteria for which the compositions and methods described herein may find beneficial use include Staphylococcus aureus (Staphylococcus aureus) , MRSA (methicillin-resistant Staphylococcus aureus), Staphylococcus epidermidis, MRSE (methicillin-resistant S. epidermidis), Mycobacterium tuberculosis, 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 faecalis, Methicillin-sensitive Enterococcus faecalis, Entero H. cloacae, Salmonella typhimurium, Proteus vulgaris, Yersinia enterocolitica, Vibrio cholere, Shigella flexneri, Vancomycin-resistant Enterococcus (VRE), Burkholderia cepacia, Francisella tularensis, Bacillus Anthracis, Yersinia pestis, Pseudomonas aeruginosa, Vancomycin-sensitive and vancomycin-resistant Enterococcus (eg, E. faecalis, E. faecium), methicillin-sensitive and methicillin-resistant Staphylococcus (eg, S. aureus, S. epidermidis) and Acinetobacter baumanee, Staphylococcus haemolyticus, Staphylococcus hominis, Enterococcus fesi , Streptococcus pyogenes, Streptococcus agalactia, Bacillus anthracis, Klebsiella pneumoniae, Proteus mirabilis, Proteus bulgaris, Yersinia enterocolitica, Stenotrohomonas maltophilia, Streptococcus pneumoniae, Penicillin, Penicillin Burkholderia cepacia, Burkholderia multivolans, Mycobacterium smegmatis and E. coli Black-backed moss.

  The practice of certain embodiments of the present invention will employ, unless otherwise indicated, conventional techniques of microbiology, molecular biology, biochemistry, cell biology, virology and immunology, which are within the skill of the art. In addition, the following reference materials are used for illustrative purposes. Such techniques are explained fully in the literature. See, 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. H. G. ins. And a. eds., 1984); Animal Cell Culture (R. Freshney ed., 1986); Perbal, A Practical Guide to Molecular Cloning (1984).

  Unless the context requires otherwise, throughout the specification and claims, the language "comprise" and variants thereof, such as "comprises" and "comprising" It is open and inclusive and must be interpreted as "including but not limited to."

  Reference to “an embodiment” or “an embodiment” or an “aspect” throughout this specification may be used to refer to a particular feature, structure or characteristic described in connection with that embodiment. 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 that embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

  As mentioned earlier, certain embodiments of the invention described herein may be directed to the agricultural, industrial, manufacturing and other uses of the described BT compounds (eg, BisEDT and / or BisBAL). With respect to formulations, the formulations may, in certain further embodiments, comprise one or more antibiotic compounds described herein, such as amikacin, ampicillin, cefazolin, cefepime, chloramphenicol, ciprofloki Sacin, clindamycin (or other lincosamide antibiotics), daptomycin (Cubicin®), doxycycline, gatifloxacin, gentamicin, imipenim, levofloxacin, linezolid (Zyvox®), minocycline, nafcillin, paromomycin , Rifampin, sulph Methoxazole, tobramycin and vancomycin; or carbapenems, cephalosporins, fluoroquinolones, glycopeptides, lincosamides, penicillinase-resistant penicillins, and / or aminopenicillins 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 require that a bacterial infection that may be biofilm-related (where bacteria that can promote biofilm formation, but biofilm is not yet detectable) is present in or on it. When administered to a plant or animal, such as a plant or animal or product, containing a bacterial infection or containing the presence of biofilms or other bacteria, as applied to a natural or artificial surface, The BT compound (s) (and, optionally, one or more antibiotics) in a suitable carrier, excipient, or diluent can be contained in an effective amount as disclosed in US Pat.

  The administration or incorporation of the BT compounds or salts thereof described herein, either in pure form or in an appropriate agricultural, manufacturing or other composition, involves the administration of an acceptable dose of a drug suitable for a similar use. It can be performed via any of the modes of incorporation. The application, contamination or administration of the composition, in a preferred embodiment, comprises contacting the composition directly with the plant or animal or product to be treated, which comprises one or more localized or broadly It may be a surface site to be distributed, generally referring to contacting a topical formulation with an acute or chronic infection site (eg, a wound site on a plant surface) surrounded by, but not limited to, intact tissue. For example, certain embodiments contemplate, as a topical application, administering a topical formulation described herein to a damaged, abraded or damaged natural or artificial surface.

  The formulation (e.g., an agricultural composition) may be a BT compound as described (e.g., RE37,793, 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 certain embodiments, as described in US Pat. Nos. 6,064,089, 5,867,098, 5,673, and 5,673,867, one or more desired antibiotics (e.g., aminoglycoside antibiotics, such as amikacin) are used separately or together with the BT compound to prepare a suitable formulation. Solid, semi-solid, gel, cream, colo, etc., depending on the intended use, in combination with a suitable vehicle, dispersant, carrier, diluent or excipient. In liquids, suspensions or liquids, or other topical forms such as powders, granules, ointments, solutions, wash, gels, pastes, plasters, liniments, bioadhesives, microsphere suspensions, and aerosol sprays , Can be formulated in a preparation.

  The compositions of these and related embodiments are formulated such that the active ingredient can be included in the compositions, and in certain preferred embodiments, the BT compounds described herein, alone or in combination with one or more Desired antibiotics (for example, carbapenem antibiotics, cephalosporin antibiotics, fluoroquinolone antibiotics, glycopeptide antibiotics, lincosamide antibiotics, penicillinase-resistant penicillin antibiotics, and aminopenicillin antibiotics) Or aminoglycoside antibiotics, such as amikacin, or rifamycin), at the desired site and, optionally, around a natural or artificial surface of a plant or animal (eg, human) subject or of a 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 are directed to the administration and / or administration of BT compounds and antibiotics to such subjects or products, such as administration that may be simultaneous or sequential and in any order. Or, although contaminants are contemplated, the invention is not so limited, and in other embodiments, explicitly contemplates separate BT compound administration routes as compared to antibiotic administration routes. That is, the antibiotic may be administered by any of the routes described herein, while the BT compound may be administered by a route independent of the route used for the antibiotic.

  The formulations described herein provide an effective amount of disinfectant (s) (and two or two antibiotics (s)) to prevent the desired site, eg, 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 and the like. And / or may be prepared to contain liposomes, micelles and / or microspheres. See, for example, U.S. Patent No. 7,205,003. For example, creams are well known in the art of pharmaceutical and cosmetic formulations, are mucous or semisolid emulsions, and are either oil-in-water or water-in-oil. Cream bases are water-washable and contain an oil phase, an emulsifier and an aqueous phase. The oil phase, also called the "inner" phase, is generally composed of petrolatum and a fatty alcohol such as cetyl alcohol or stearyl alcohol. The aqueous phase is usually, but not necessarily, greater in volume than the oil phase and generally contains a humectant. Emulsifiers in cream formulations are generally non-ionic, anionic, cationic or amphoteric surfactants.

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

  Gels are semi-solid, suspension-type systems. Single-phase gels contain organic macromolecules that are typically aqueous but preferably substantially homogeneously distributed throughout a carrier liquid that also contains alcohol, and any oils. Preferred "organic macromolecules" or gelling agents may be chemically cross-linked polymers, such as cross-linked acrylic acid polymers, for example "carbomers" of polymers, such as carboxypolyalkylenes, which are available from 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 Propyl methylcellulose phthalate and methylcellulose; gums such as tragacanth 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 trituration, mechanical mixing or stirring, or a combination thereof.

  Ointments, also well known in the art, are semisolid preparations, typically based on petrolatum or other petroleum derivatives. The particular ointment base employed will provide a number of desired characteristics, such as emollientity, as will be appreciated by those skilled in the art. As with other carriers or vehicles, an ointment base must be inert, stable, nonirritating, and nonsensitizing. As described in Remington: The Science and Practice of Pharmacy, 19th Ed (Easton, Pa .: Mack Publishing Co., 1995), p1399-1404, ointment bases are classified into oily bases; emulsifying 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 absorbable ointment bases, contain little or no water and include, for example, hydroxystearin sulfate, anhydrous lanolin, and hydrophilic petrolatum. The ointment base of the emulsion is either a water-in-oil (W / O) emulsion or an oil-in-water (O / W) emulsion and includes, 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 predecessor).

  Pastes are semi-solid dosage forms, the active agent being suspended in a suitable base. The paste is fractionated into a fat paste or a paste made of a single-phase aqueous gel, depending on the nature of the base. The base in the fat paste is generally petrolatum or hydrophilic petrolatum or the like. Pastes made with single-phase aqueous gels generally incorporate carboxymethylcellulose or the like as a base.

  The formulation may be prepared using liposomes, micelles and microspheres. Liposomes are microscopic vesicles having one (unilamellar) or multiple (multilamellar) lipid walls comprising a lipid bilayer, as used herein, one or more of the formulations described herein. Components, such as disinfectants, or specific carriers or excipients, may be encapsulated and / or adsorbed on the lipid membrane surface. The liposome preparations herein include cationic (positive charges), anionic (negative charges), and neutral preparations. Anionic liposomes are readily available. For example, N [1- (2,3-dioleyloxy) propyl] -N, N, N-triethylammonium (DOTMA) liposome is commercially available under the trade name Lipofectin (registered trademark) (GIBCO BRL, Grand Island, NY). ). Similarly, anionic and neutral liposomes are readily available, for example, from Avanti Polar Lipids (Birmingham, AL), or can be readily prepared using readily available materials. Such materials include phosphatidylcholine, cholesterol, phosphatidylethanolamine, dioleylphosphatidylcholine (DOPC), dioleylphosphatidylglycerol (DOPG), and dioleylphosphatidylethanolamine (DOPE), among others. 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 such that the polar head group forms an outer spherical shell and hydrophobic hydrocarbon chains are arranged toward the center of the sphere to form a core. Is known in the art. Since micelles are formed in an aqueous solution containing a sufficiently high concentration of a surfactant, micelles are obtained spontaneously. Surfactants useful for forming micelles include, but are not limited to, potassium laurate, sodium octanesulfonate, sodium decanesulfonate, sodium dodecanesulfonate, 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 formulations of the present invention. They are generally, but not necessarily, formed from lipids, preferably charged lipids, such as phospholipids. 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, solvents containing relatively small amounts 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 and the like; polyethylene glycol and its esters such as polyethylene glycol monolaurate (PEGML; see, eg, US Pat. 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 C10 MSO, may be used, but are less preferred.

  Certain skin penetration enhancers are typically referred to as "plasticizing" enhancers, a lipophilic co-enhancer, i.e., a molecular weight in the range of about 150-1000 daltons, about 1 weight. %, Preferably less than about 0.5% by weight, most preferably less than about 0.2% by weight of an enhancer having water solubility. The Hildebrand solubility parameter of the plasticizing 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 Milli-start. Fatty alcohols include, for example, stearyl alcohol and oleyl alcohol. 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 topical drug delivery and / or are described in the relevant literature. See, for example, Percutaneous Penetration Enhancers eds. Smith et al. (CRC Press, Boca Raton, FL, 1995).

  In addition to those previously identified, various other additives may be included in a topical formulation according to certain embodiments of the present invention. These include, but are not limited to, antioxidants, astringents, fragrances, preservatives, emollients, pigments, dyes, hygroscopic agents, propellants, and sunscreens, as well as cosmetics, chemicals, Or other types of materials that are otherwise desirable. Typical examples of optional additives included in the formulations of certain embodiments of the invention are: preservatives, such as sorbate; solvents, such as isopropanol and propylene glycol; astringents, such as Menthol and ethanol; emollients such as polyalkylenemethylglucoside; hygroscopic agents such as glycerin; emulsifiers such as glycerol stearate, PEG-100 stearate, polyglyceryl-3 hydroxylauryl ether, and polysorbate 60; sorbitol and other polyhydroxy alcohols Sunscreens such as octyl methoxycinnamate (commercially available as Parsol MCX) and butyl methoxybenzoylmethane (commercially available as Parsol 1789) Antioxidants such as ascorbic acid (vitamin C), α-tocopherol (vitamin E), β-tocopherol, γ-tocopherol, δ-tocopherol, ε-tocopherol, Δ1-tocopherol, Δ2-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 oils (eg, perhydrosqualene), mineral oils, synthetic oils, silicone oils or waxes (eg, cyclomethicone and dimethicone), fluorinated oils (Generally perfluoropolyate ), Fatty alcohols (e.g., cetyl alcohol), and waxes (e.g., beeswax, carnauba wax, and paraffin wax); skin-feel modifiers; and thickeners and structures, e.g. Crosslinked carboxypolyalkylenes which may be obtained commercially as swellable clays and under the trade name Carbopol®.

  Other additives include, for 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 are beneficial to include include, for example, α-hydroxy acids, α-keto acids, polymeric hydroxy acids, humectants, collagen, marine extracts, and antioxidants such as ascorbic acid (vitamin C), Alpha-tocopherol (vitamin E) or other tocopherols, such as those described above, and retinol (vitamin A), and / or appropriate salts, esters, amides, or other derivatives thereof, may be present. Additional agents include those capable of improving oxygen supply in living tissue, such as those described in WO 94/00998 and WO 94/00109. A sunscreen may be included.

  The formulations of certain embodiments of the present invention may include conventional additives such as opacifiers, fragrances, colorants, gelling agents, thickeners, stabilizers, surfactants, and the like. Other agents may be added to prevent spoilage during storage, ie, to inhibit the growth of microorganisms such as yeasts and molds, for example, antimicrobial agents. 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 can be 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 effective amounts of one or more additional active agents appropriate to the particular mode of administration or incorporation.

  A pharmaceutically acceptable carrier may be incorporated into the topical formulation of a particular embodiment of the 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, daritol and iditol (6-carbon), isomaltol, maltitol, lactitol and polyglycitol, hydrocarbon oils, fats, waxes, 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 can be treated 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 periodically to everything that requires. The frequency of treatment will depend 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, eg, amikacin or other antibiotics), delivery of the active ingredient. Depends on the effectiveness of the vehicle used to remove the formulation, as well as the ease with which the formulation can be removed due to environmental factors (eg, physical contact with other substances or objects, precipitation, wind, temperature).

  Typical concentrations of the active agent, such as the BT compound 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%. And more preferably in the range of about 0.1 to 2.0%. As a representative example, the compositions of these embodiments of the invention may be applied to natural or artificial surfaces at a rate of about 1.0 mg / cm2 to about 20.0 mg / cm2. 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, Gels, hydroalcoholic solutions, liposomes, lotions, microemulsions, ointments, oils, organic solvents, polyols, polymers, powders, salts, silicone derivatives, and waxes. The formulation may include, for example, chelating agents, conditioning agents, emollients, excipients, humectants, protectants, thickeners, or UV absorbers. 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 divalent cation metals such as Ca2 +, Mn2 +, or Mg2 +. Examples of chelating agents include, but are not limited to, EDTA, disodium EDTA, EGTA, citric acid, and dicarboxylic acids.

  Conditioning agents may 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, albumen, 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 Erol, 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 factors, ergocalciferol, ergosterol, ethylhexyl PCA, fibronectin, folate, 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 salts, 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, stearamidopropyl betaine , Stearyl palmitate, tocopherol, tocopheryl acetate, tocopheryl linoleate, ubiquinone, Vitis vinifera (grape) seed oil, wheat flour Noic acid, xanthan gum, and zinc gluconate. As will 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, including, but not limited to, acetylated lanolin, acetylated lanolin alcohol, acrylate / C10-30 alkyl acrylate crosspolymer, acrylate copolymer, alanine, seaweed extract, aloe barbadensis extract or gel, Altea officinalis extract, starch aluminum octenyl succinate, aluminum stearate, apricot (Prunus Armenica) Kernel oil, arginine, arginine aspartate, arnica montana extract, ascorbic acid, ascorbyl palmitate, aspartic acid, avocado (Persia gratissima) oil, barium sulfate, barriers 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, squid (Ruscus Aculeatus) extract, Butylene glycol, Calendula officinalis extract, Calendula officinalis oil, Calendula (Euphorbia cerifera) wax, canola oil, caprylic / capric triglyceride, cardamom (Eletaria cardamomum) oil, carnauba ( Copernicia cerifera) wax, carrageenan (condras crispas), carrot (daucas carota sativa) oil, castor (lisinas communis) oil, sera De, ceresin, ceteareth-5, ceteareth-12, ceteareth-20, cetearyl octanoate, 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 cacao) butter, coco-caprylic / capric acid, coconut (cocos nucifera) oil, collagen, collagen amino acids, corn (g Mace) Oil, fatty acid, decyl oleate, dextrin, diazolidinyl urea, dimethicone copolyol, dimethiconol, dioctyl adipate, dioctyl succinate, hexacaprylic acid / dihexacaprate Nantaerythritil, DMDM hydantoin, DNA, erythritol, ethoxydiglycol, ethyl linoleate, eucalyptus globulus oil, evening primrose (Oenocera biennis) oil, fatty acid, fructose, gelatin, wild geranium oil, glucosamine, glutamate glucose, glutamic acid, glyceres-26 , Glycerin, glycerol, glyceryl distearate, glyceryl hydroxystearate, glyceryl laurate, glyceryl linoleate, glyceryl myristate, glyceryl oleate, glyceryl stearate, glyceryl stearate SE, glycine, glycol stearate, glycol stearate SE, Glycosaminoglycan, grape (Vitis Vinichella) seed oil, hazel (Colliras Amelie) Cana) nut oil, hazel (Colliras 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 butyl carbamate, isocetyl stearate, stearoyl isocetyl stearate, isodecyl oleate, isopropyl isostearate, lano Isopropyl acetate, isopropyl myristate, isopropyl palmitate, isopropyl stearate, isostearamide DEA, isostearic acid, isostearyl lactate, isostearyl neopentanate, jasmine (Jasminum officinale) oil, jojoba (Baxus tinensis) oil, kelp , Kukui (Aleurites molkana) nut oil, Lactamide MEA, Runes-16, Runes-10 acetate, Lanolin, Lanolinic acid, Lanolin alcohol, Lanolin oil, Lanolin wax, Lavender (lavandula angstifolia) oil, Lecithin, Lemon ( Citrus Medica Rimonam) oil, linoleic acid, linolenic acid, macadamia ternifolia nut oil, magnesium stearate, magnesium sulfate, maltitol, mosquito Millet (chamomile recutita) oil, methylglucose sesquistearate, methylsilanol PCA, microcrystalline wax, mineral oil, mink oil, Mortierella oil, myristyl myristate, myristyl myristate, myristyl propionate, neopentyl dicaprylate / dicaprate Glycol, octyldodecanol, octyldodecyl myristate, octyldodecyl stearoyl stearate, octyl hydroxystearate, octyl palmitate, octyl salicylate, octyl stearate, oleic acid, olive (olea europaea) oil, orange (citrus aurantium)・ Dursis) oil, coconut (Elais Guinensis) oil, palmitic acid, pantethine, 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 stearate glyceryl, 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-stearate 32, PEG-40 stearate, PEG-50 stearate, PEG-stearate 100, PEG-150 stearate, pentadecalactone, peppermint (mentha piperita) oil, petrolatum, phospholipid, polyaminosugar 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 diperargonate, propylene glycol laurate, propylene glycol stearate, SE propylene glycol stearate, PVP, pyridoxine dipalmitate, quatanium- 5. Quotanium-18 hectorite, quatanium-22, retinol, retinyl palmitate, rice (Oriza sativa) nuka oil, RNA, rosemary (Rosmarinus officinalis) oil, rose oil, safflower (Casamas tinctorias) oil , Sage (Salvia officinalis) oil, salicylic acid, sandalwood (Santalam album) oil, serine, serum protein, sesame (Sesamemu indicum) oil, shea butter (butyrospermparky), silk powder, sodium chondroitin sulfate, DNA Sodium, sodium hyaluronate, sodium lactate, sodium palmitate, sodium PCA, sodium polyglutamate, sodium stearate, water-soluble collagen, sorbic acid, sorbitan laurate, Sorbitan inmate, sorbitan palmitate, sorbitan sesquioleate, sorbitan stearate, sorbitol, soybean (glycin soya) oil, sphingolipid, squalane, squalene, stearamide MEA stearate, stearic acid, stearoxydimethicone, stearoxytrimethylsilane, Stearyl alcohol, stearyl glycyrrhetinate, stearyl heptanoate, stearyl stearate, sunflower (Helianthus anus) seed oil, sweet almond (Prunus amygdalus dursis) oil, synthetic beeswax, tocopherol, tocopheryl acetate, tocopheryl linoleate, tribehenine, Tridecyl neopentanoate, tridecyl stearate, triethanolamine, tristearin, urea, vegetable oil, water, Wax, wheat (Triticum bulgare) germ oil, and ylang-ylang (Cananga odorata) oil.

  Surfactants can also be included, if desired, in certain formulations contemplated herein, including cationic, anionic, zwitterionic or non-ionic surfactants, or their surfactants. It can be selected from any natural or synthetic surfactant suitable for use in a cosmetic composition, such as a mixture (Rosen, M., "Surfactants and Interfacial Phenomena," Second Edition, John Wiley & Sons, NY. , 1988, Chapter 1 p431). 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 Long chain amines can be mentioned, 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; And salts of perfluorocarboxylic acid. Examples of zwitterionic surfactants include cocamidopropylhydroxysultaine (CAPHS) and others that are pH sensitive and require special care in designing the optimal pH of the formulation (ie, , Alkylaminopropionic acid, imidazoline carboxylate, and betaine) or those that are not pH sensitive (eg, sulfobetaine, sultaine). Examples of nonionic surfactants include, but are not limited to, alkylphenol ethoxylates, alcohol ethoxylates, polyoxyethylenated polyoxypropylene glycols, polyoxyethylenated mercaptans, long chain carboxylic esters, alkanols Mention may be made of amides, tertiary acetylene glycols, polyoxyethylenated silicones, N-alkylpyrrolidones, and alkyl polyglycosidases. Agents such as wetting agents, mineral oils or other surfactants such as non-ionic detergents or one or more of the PLURONIC® series (BASF, Mt Olive, NJ) may also be used, eg, by non-limiting theory. It may be included in order to prevent aggregation of the BT fine particles in the fine particle suspension. Any combination of surfactants is acceptable. Particular embodiments include at least one anionic surfactant and at least one cationic surfactant that are compatible, ie, do not form a complex that appreciably precipitates 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 copolymers, agarose, amylopectin, bentonite, calcium alginate, calcium carboxymethylcellulose, carbomer, carboxymethyl chitin, 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, sodium carrageenan, 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 sunscreen agents or UV absorbers. If 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, sinoxate, DEA-methoxycinnamic acid, diisopropyl methylcinnamate, ethyldihydroxypropyl PABA, diisopropylcinnamic acid Ethyl, ethyl methoxycinnamate, ethyl PABA, ethyl urocanate, glyceryl octanoate dimethoxycinnamate, glyceryl PABA, glycosalicylate 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 protectors 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 or about the following pH range: 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 From about pH 10, from about pH 3 to about pH 9, from about pH 3 to about pH 8, and from about pH 3 to about pH 8.5. Most preferably, the pH is between about pH 7 and about pH 8. One skilled in the art may add appropriate pH adjusting components to the compositions of the present invention to adjust the pH to an acceptable range. If it is recognized that the pH may fluctuate from the original value depending on the composition of the formulation and the storage conditions, the specific pH with “about” may be the specific pH measured at any given time. Those of skill in the art will understand that they include formulations that can be up or down by 0.7, 0.6, 0.5, 0.4, 0.3, 0.2 or 0.1 pH units above or below the critical value. There will be.

  Creams, lotions, gels, ointments, pastes and the like may be spread on the affected surface and gently rubbed in. The solution may be applied in the same manner, but more typically will be applied using a dropper, swab, etc., and will be applied carefully to the affected area. The 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 of skill in the art can readily determine the appropriate amount of formulation to be administered, the mode 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 one or two or more times a week to 1, 2, 3, 4 or more times a day. Is contemplated.

  As also discussed above, the BT formulations useful herein may also contain an acceptable carrier, including any suitable diluents or excipients, and as such, may be used to form the composition. It includes any agent that is not harmful to the subject to be administered (eg, a plant or animal (including a human)) and can be administered without undue toxicity. Acceptable carriers include, but are not limited to, liquids such as water, saline, glycerol and ethanol, as well as 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 provided in REMINGTON'S PHARMACEUTICAL SCIENCES (Mack Pub. Co., NJ, latest edition).

  A BT formulation can include an agent that binds to the BT compound and thereby assists in delivering or retaining it at a desired site on a 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, depending on various factors, such as the nature of the site of delivery (if relevant), the activity of the particular BT compound used (aminoglycoside antibiotics, Eg, with or without antibiotic formulations such as amikacin); metabolic stability and duration of action of the compound; (plant or animal, including human) state of the subject or product; mode of administration and time; excretion Rate; the rate of loss of the BT compound in the normal course of activity that the subject or product undergoes; and will vary depending on other factors. Generally, a therapeutically effective daily dose will be from about 0.001 mg / kg (i.e., 0.07 mg) to about 100 mg / kg (i.e., 7.0 g) (for a 70 kg mammal); , (In a 70 kg mammal) from about 0.01 mg / kg (i.e., 7 mg) to about 50 mg / kg (i.e., 3.5 g); more preferably, a therapeutically effective dose is (in a 70 kg mammal). From about 1 mg / kg (ie, 70 mg) to about 25 mg / kg (ie, 1.75 g). Effective doses of plants can be expected to be about 10, 20, 50 or 75% or more lower.

  The effective dose ranges provided herein are not limiting and represent the preferred dose range. However, the most preferred doses will be tailored to each subject, as will be understood and determinable by those skilled in the relevant art (eg, Berkov et al., Eds., The Merk Manual 16th edition, Merk and). Co., Rahway, NJ, 1992; Avery's Drug Treatment: Principles and Practice of Clinical Pharmacology and Therapeutics, 3rd edition, ADIS Press, Ltd., Williams and Wilkins, Baltimore, MD. 'S Pharmaceutical Sciences, 18th edition, Mack Publishing Co., Easton, PA (1990); Katzung, Basic and Clinical Pharmacology, Appl. ) Reference).

If desired, the total dose required for each treatment can be administered in the same day as a multiple dose or a single dose. Certain preferred embodiments contemplate applying a single application of the BT formulation per day, per week, per 10 days, per 14 days or longer. Generally, 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 plant and agricultural products

  Certain embodiments disclosed herein relate to compositions and methods for protecting plants and flowers from microbial infection and infestation, such as biofilms, reducing mortality and increasing product life.

  According to certain embodiments described herein, for example, those summarized above, a method for protecting a plant against a bacterial, fungal, or viral pathogen, comprising: Prevention of plant infection by viral pathogens, (ii) inhibition of cell viability or cell growth of substantially all planktonic cells of bacterial, fungal or viral pathogens, (iii) biofilm formation by bacterial, fungal or viral pathogens And (iv) inhibiting the biofilm viability or biofilm growth of substantially all biofilm-type cells of the bacterial, fungal or viral pathogen in sufficient conditions and times. Contacting a plant with an effective amount of a BT composition, wherein the BT composition comprises fine particles of 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 one embodiment, the bacterial pathogen comprises an Erwinia amylobora cell, and in some embodiments, the bacterial pathogen is an Erwinia amilobolla, Xanthomonas campestris pathogen dieffenbachiae, Pseudomonas syringae, Xylella fastidiosa; Xylophilus amperinus; -Selected from Fructicola, Pantoea stewartii subsp. Stewartii, Ralstonia solanacearum and Clavibactor michiganensis subsp. Sepedonicus. In certain embodiments, the bacterial pathogen exhibits antibiotic resistance, and in certain other embodiments, the bacterial pathogen exhibits streptomycin resistance. In certain embodiments, the plant is a food crop plant, which is, in certain further embodiments, a fruit tree, which, in further embodiments, is selected from apple, pear, peach, nectarine, plum, apricot trees. In one embodiment, the food crop plant is a banana tree of Musa. In certain other embodiments, the food crop plant is a plant selected from a tuberous root plant, a legume plant, and a grain plant. In a further embodiment, the tuberous plant is selected from potato (potato) and sweet potato (sweet potato).

  In some embodiments, the contacting step is performed one or more times. In certain embodiments, at least one step of contacting comprises one of spraying the plant, dipping the plant, coating the plant, and painting the plant. In certain embodiments, at least one step of the contacting is performed at a flower bud, shoot or growth site of the plant, or at other plant parts, such as roots, bulbs, stems, leaves, branches, vines, creeping branches, It is carried out on or in buds, flowers or parts thereof, shoots, fruits, seeds, pods 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 is BisBAL, BisEDT, Bis-dimercaprol, Bis-DTT, Bis-2-mercaptoethanol, Bis-DTE, Bis-Pyr, Bis-Ery, Bis-Tol, Bis-BDT. , 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 method, the method comprises synergistically or synergistically enhancing the antibiotic with the plant in association with the step of contacting the plant with the BT composition, simultaneously or sequentially, and in any order. Further contacting with. In certain 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. Including antibiotics selected from substances. In certain embodiments, the synergistic or potentiating antibiotic is an aminoglycoside antibiotic selected from amikacin, arbekacin, gentamicin, kanamycin, neomycin, netilmycin, paromomycin, rhodostreptomycin, streptomycin, tobramycin, and apramycin.

In another embodiment, a method for overcoming antibiotic resistance in a plant in which an antibiotic resistant bacterial plant pathogen is present, wherein the method comprises: (a) one or more of the following: Contacting the plant with an effective amount of the BT composition under conditions and for a time sufficient to: (i) prevent infection of the plant by the antibiotic-resistant bacterial pathogen; (ii) substantially all of the antibiotic-resistant bacterial pathogen Inhibition of cell viability or cell growth of planktonic cells, (iii) inhibition of biofilm formation by antibiotic-resistant bacterial pathogens, and (iv) biochemistry of substantially all biofilm-type cells of antibiotic-resistant bacterial pathogens Inhibition of film viability or biofilm growth (where the BT composition comprises a substantially monodisperse suspension of microparticles comprising a BT compound; Has a volume average diameter of about 0.5 μm to about 10 μm), and (b) in connection with the step of 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.
Disinfectant based on bismuth-thiol (BT)

  As noted above, a number of natural products (eg, antibiotics) and synthetic chemicals having antimicrobial (antibacterial, antiviral, antifungal) properties, particularly antibacterial properties, are known in the art and include at least one The part has a chemical structure as well as an antimicrobial action, for example the ability to kill microorganisms ("killing" action, e.g. bactericidal properties), the ability to stop and impair microbial growth ("static" action, e.g. Or the ability to interfere with microbial functions, such as colonizing or infecting a site, bacterial secretion of exopolysaccharide, and / or conversion of a planktonic population to a biofilm population, or expansion of biofilm formation. Are characterized by 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 ability, effective concentrations, and host tissue. Including the dangers of toxicity to, are discussed above and in US Patent No. 6,582,719.

  Bismuth-thiol (BT), as well as related thiol compounds in which bismuth is replaced by different Group V metals (eg, arsenic, antimony) are discussed above. Beneficial particulate BT compositions having a volume average diameter of about 0.5 μm to about 10 μm Compositions and methods for particulates are also discussed herein. Accordingly, certain exemplary embodiments relate to the use of an antimicrobial agent described herein, eg, an anti-biofilm agent, to treat or prevent infections and biofilms in plants. The agent is typically present in alkaline form in a composition containing one or more particulate bismuth-thiols in 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 components, such as other compatible bactericides, and in certain preferred embodiments described herein. Including synergistic or potentiating antibiotics.

  Target crops to be protected within certain contemplated but non-limiting embodiments include, for example, the following plant species: Cereals (eg, wheat, barley, rye, oats, rice, sorghum and related Crops), beets (eg, sugar beet and forage beets), pears, drupes and soft fruits (eg, apples, pears, plums, peaches, almonds, cherries, strawberries, raspberries and blackberries), legumes (eg, Kidney beans, lentils, peas, soybeans), oily plants (eg, rape, mustard, poppy, olives, sunflowers, coconut, castor bean plants, cocoa beans, peanuts), cucumber plants (eg, cucumber, squash, melon), Textile plants (eg, cotton, flax, hemp, jute), citrus fruits (eg, Range, lemon, grapefruit, mandarin orange), vegetables (eg, spinach, lettuce, asparagus, cabbage, carrot, onion, tomato, potato, paprika), camphor (eg, avocado, cinnamon, camphor), and other plants For example, corn, tobacco, nuts, coffee, sugar cane, tea, vines, hops, bananas and natural rubber plants, and ornamental plants (Asteraceae), such as floriculture plants and cut flowers thereof. Thus, certain embodiments provide for contacting a crop item with a composition comprising one or more of the particulate BT compounds as provided herein, such that a target crop item to be harvested, such as a cut flower or Extending the life of the target crop-derived food (e.g., fruits, vegetables, grains, seeds, etc.) (e.g., if the item is commercially available compared to a control group that is not contacted with the particulate BT according to the present invention). To prolong a period of nutritional and / or aesthetic benefit that is statistically significant).

  The effective concentration of particulate BT as described herein for use in these and related embodiments depends on a number of factors, such as BT, pH, temperature, molar ratio of BT components, and from harmful microorganisms. Depends on what is selected. Efficacy also depends on whether prevention of infection or treatment of existing infection (biofilm) is a goal of a particular application. A protective 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 as high as 8 μg / ml or more, depending on the specific particulate BT compound (s). In one exemplary embodiment, the particulate Bis pyrithione (BisPyr) is provided in a 5: 1 molar ratio (bismuth to pyrithione) for application to plants. In another embodiment, the 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 fortified and powerful defenses. 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 baking soda (sodium bicarbonate) or other alkaline substance (s) (eg, potassium bicarbonate, calcium carbonate) to the particulate BT formulation adds to the antimicrobial activity of BT. Or may enhance it. Other ingredients in particulate BT formulations 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 bisphenolic 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; Essential oils of Lultam, A-dec ICX, Coleus Forskoli; silver or silver colloid antimicrobial agents; antimicrobial agents based on tin or copper; chlorine or bromine oxidants, manuka oil; oregano; thyme; rosemary; Other herbal extracts; and grapefruit seed extracts; anti-inflammatory or antioxidants such as ibuprofen, flurbiprofen, aspirin, indomethacin, aloe vera, turmeric, olive leaf extract, clove, panthenol, retinol, omega- Trifatty acids, gamma linolenic acid (GLA), green tea, ginger, grape seeds and the like; pharmaceutically acceptable carriers such as starch, sucrose, water or water / alcohols, DMSO and the like; surfactants such as anionic, non-ionic Ionic, cationic and zwitterionic On- or amphoteric surfactants or saponins from plant material (see, eg, US Pat. No. 6,485,711); buffers and salts; and other components 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 use and for use with plants may, in certain embodiments, be additive, enhancing or synergistic, as described herein or in liposome or nanoparticle form. These and optionally other agents that produce an effect may be combined to enhance activity and delivery. Certain embodiments explicitly exclude particulate BT formulations that include 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. Particulate BT containing carriers, excipients, or other additives that promote adhesion of the formulation to the surface (eg, glucose, starch, citric acid, carrier oil, emulsions, dispersions, surfactants, etc.) May also be prepared.

  In other contemplated embodiments, a particulate BT formulation for use as an anti-biofilm agent on a plant or crop 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 Patent Nos. 7,427,408 and 6,445,031) help control biofilms. These anti-biofilm agents in combination with the particulate BT described herein can be delivered by foliar spray for inhibition of bacterial biofilm development and for treatment of preformed biofilms. In another embodiment, these anti-biofilm agents are contained within biodegradable microparticles for controlled release and / or in liposome form with other antimicrobial agents.

  Thus, the microparticles BT described in the present invention, according to certain embodiments, can be used with other current technologies to improve the anti-biofilm effect. The particulate BT of the present invention may synergize or enhance the activity of the antibiotics streptomycin and / or gentamicin against certain plant pathogens. Streptomycin does not kill bacteria, but instead suppresses their growth, thus reducing the rate at which the stigmas of flowers colonize, thereby reducing the subsequent growth of bacteria within the nectary gland (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). Additional benefits may result from the use of an active spray adjuvant (eg Regulaid ™) that improves the coverage and penetration of this antibiotic sufficient to safely use reduced amounts of streptomycin.

  The particulate BT of the present invention can be combined with any of the active ingredients currently in use to combat agricultural and plant microbial pathogens, such as any of oxidants, chelators (eg, iron chelators), fungicides and disinfectants. Preferred combinations may be additive or may be potentiating or synergistic in accordance with the present disclosure with respect to their anti-biofilm effects. Some embodiments include particulate BT compositions that are formulated to be hydrophobic to enhance BT retention at the surface, for example, by using a hydrophobic thiol (eg, thiochlorophenol) to provide enhanced adhesion properties. Intended thing. 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 halogenated hydrocarbon propellants, dispersing oils, or as a dry powder. Aqueous formulations can be aerosolized by a liquid nebulizer using hydraulic or ultrasonic atomization. Propellant-based systems may employ a suitable pressurized dispenser. Dry powders 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 choosing the appropriate equipment.

  Throughout this specification, unless otherwise required, the words "comprise", "comprises" and "comprising" are inclusive of the referenced step or elements, or steps or elements. It should be understood, however, that this does not exclude any other steps or elements or steps or elements. “Consisting of” means including the matter following the idiom “consisting of”, and is limited thereto. That is, the phrase "consisting of" indicates that the listed elements are required or required, and that no other elements can be present. "Consisting essentially of" means including any element listed after the phrase, and does not interfere with or contribute to the activity or action specified in the disclosure regarding the listed element. Limited to elements. That is, the phrase "consisting essentially of" requires or requires the listed elements, but does not require other elements, depending on whether they affect 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 referents unless the context clearly dictates otherwise. The term “about” or “approximately”, as used in certain embodiments herein, precedes a numerical value by its value plus or minus 5%, 6%, 7%, 8%, or 9%. Indicates the range. In other embodiments, the term "about" or "approximately," if present before a numerical value, indicates the value plus or minus 10%, 11%, 12%, 13%, or 14%. In yet another embodiment, the term “about” or “approximately”, if present before a numerical value, plus or minus that range of 15%, 16%, 17%, 18%, 19%, or 20%. Is shown.

References: 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. Feazel LM, Baumartner LK, Peterson KL, et al. Opportunistic pathsogens enriched in showerhead biofilms. PNAS 2009 (epub ahead of print). Seesey GG, Lewowski Z, Flemming 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 detachment method. Congres 2008; 220: 290-4. Ozdamar et al. , Retina 1999; 19: 122-6. Piccirillo 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.S. 2000. Cytokine modulation by bismuth-ethanidithiol 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, Scherble J. Reloadable antimicrobial coatings based on amphiphilic silicone 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 Technology 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, hS. 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 Expl 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. See 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 the 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: 7687-7922. 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 not limitation.

Example 1
Preparation of BT Compound The following BT compound was 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 synthetic protocol described below for BisEDT. Shown are the atomic ratios for a single bismuth atom as a comparison, based on the stoichiometric 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 agents (eg, Bi: thiol1 / thiol2; 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)

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

  An excess of room temperature 5% aqueous HNO3 (11.4 L) in a 15 L polypropylene carboy was added to an aqueous Bi (NO3) 3 solution (43% Bi (NO3) 3 (w / w), 5% nitric acid (w / w). ), 52% (w / w) water, 0.331 L (~ 0.575 mol) of Shepherd Chemical Co., Cincinnati, OH, product number 2362; δ-1.6 g / mL) is slowly added dropwise with stirring. Then, absolute ethanol (4 L) was added slowly. A small amount of white precipitate formed, but dissolved with continued stirring. Separately, using a 60 mL syringe, 72.19 mL (0.863 mol) of 1,2-ethanedithiol (CAS 540-63-6) was added to 1.5 L of absolute ethanol, and then stirred for 5 minutes. , 1,2-ethanedithiol in ethanolic solution (〜1.56 L, ０．５0.55 M). Thereafter, the 1,2-ethanedithiol / EtOH reagent was slowly added dropwise to the aqueous Bi (NO3) 3 / HNO3 solution over 5 hours, and the mixture was continuously stirred overnight. The product formed was allowed to settle out 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 successively washed three times with 500 mL each of ethanol, USP water and acetone to remove BisEDT (694.51 g / mol) from yellow amorphous. Powdery solid. The product was placed in a 500 mL brown glass bottle and dried over CaCl2 under high vacuum for 48 hours. The recovered material (yield ~ 200 g) emitted a odor characteristic of thiols. The crude product was redissolved in 750 mL of absolute ethanol, stirred for 30 minutes, then filtered and washed three times with 50 mL of ethanol, twice with 50 mL of acetone, and again with 500 mL of acetone. The rewashed powder was triturated in 1 M NaOH (500 mL), filtered and washed three times with 220 mL of water, twice with 50 mL of ethanol, and once with 400 mL of acetone to obtain 156.74 g of purified BisEDT. . The next batch, prepared in essentially the same manner, resulted in a yield of about 78.91%.

  The product was analyzed by analysis of data from 1H and 13C nuclear magnetic resonance (NMR), infrared spectroscopy (IR), ultraviolet spectroscopy (UV), mass spectroscopy (MS) and elemental analysis to give the formula shown earlier It was characterized as having the structure of I. A method of HPLC was developed to determine the chemical purity of BisEDT, whereby samples were prepared 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 UV detector monitoring at 265 nm (λmax), equipped with a 2 μL injection volume and a YMC Pack PVC Sil NP (250 × 4.6 mm id) analytical column (Waters) Using a 2695 chromatograph, isocratic HPLC elution at 1 mL / min was performed at ambient temperature with 0.1% formic acid in acetone: water (9: 1) as mobile phase and a single peak was detected. It showed a chemical purity of 100 ± 01%. Elemental analysis was consistent with the structure of formula (I).

  The dry particulate material was characterized to evaluate the particle size characteristics. Briefly, microparticles were resuspended in 2% Pluronic® F-68 (BASF, Mt. Olive, NJ) and the suspension sonicated for 10 minutes in a standard water bath sonicator. After that, analysis was carried out using a Nanosizer / Zetasizer Nano-S particle analyzer (model ZEN1600 (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 with all detectable events occurring between 0.6 micron to 4 micron volume mean diameter (VMD) and peaking at about 1.3 micron Had a VMD. In contrast, when BisEDT is prepared by prior art methods (Domenico et al., 1997 Antimicrob. Agents Chemother. 41 (8): 1697-1703), the majority of the particles are heterodisperse and significantly larger in size, resulting in a VMD Characterization based on

Example 2
Colony Biofilm Model of Chronic Wound Infection: Inhibition by BT Compounds The bacteria present in chronic wounds are essentially according to the described method for adopting the 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 Biotchnol. Prog. 12: 316; Zheng et al., 2002; The effect of BT on biofilms was examined for the effect on bacterial cell survival using biofilms. Inspected.

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

  Bacterial biofilm colonies were isolated essentially as described (Anderl et al., 2003 Antimicrob Agents Chemother 47: 1251-56; Walters et al., 2003 Antimicrob Agents Chemother 47: 317. Biol. Prog. 12: 316; Zheng et al., 2002 Antimicrob Agents Chemother 46: 900), grown on microporous membranes on agar plates. Colony biofilms exhibit many of the well-known properties of other biofilm models, for example, they were composed of cells that were densely aggregated in a hyperhydrated 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. Med Microbiol 46;: 425), bacteria in colony biofilms were also observed to exhibit significantly reduced antimicrobial susceptibility, as quantified in higher performance in vitro biofilm reactors. Colony biofilms occurred immediately and reproducibly in large numbers. According to non-limiting theory, this colony biofilm model shares some of the characteristics of the infected wound, and bacteria are air-conditioned with nutrients supplied from beneath the biofilm and a minimal amount of fluid. Growing at the interface. Various sources of nutrients were used to culture the colony biofilm, such as blood agar, which would mimic in vivo nutritional conditions.

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

  The colony biofilm is supplemented with candidate antimicrobial treatments to determine sensitivity to antimicrobial agents (eg, BT compounds such as BT compound combinations; antibiotics; and BT compound-antibiotic combinations). And transferred to the agar plate. If the duration of exposure to the 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 cases, the sample had to be processed briefly with a tissue homogenizer to break down cell aggregates. The resulting cell suspension was then serially diluted and allowed to settle, and the surviving bacteria were enumerated and reported as colony forming units per unit area (CFU). Survival data was analyzed using log10 transformation.

  Each type of bacterial biofilm colony culture (Pseudomonas aeruginosa, PA; methicillin-resistant Staphylococcus aureus, MRSA or SA) was tested for 5 antibiotics and 13 BT compounds. The antimicrobial agents tested for PA were BT, referred to herein as BisEDT and Compounds 2B, 4, 5, 6, 8-2, 9, 10, 11 and 15 (see Table 1), and antibiotics. Tobramycin, amikacin, imipenim, cefazolin, and ciprofloxane. The antimicrobial agents tested for SA include BisEDT and BT, designated compounds 2B, 4, 5, 6, 8-2, 9, 10 and 11 (see Table 1), and the antibiotics rifampicin, daptomycin, minocycline, Ampicillin, and vancomycin. Antibiotics were tested at approximately 10-400 times the minimum inhibitory concentration (MIC) according to established microbiological methodologies, as described earlier in the Brief Description of the Drawings.

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

Example 3
Inhibition of chronic wound infections by the drip flow biofilm model BT compound Drip flow biofilm is a recognized and reliable model for forming bacterial biofilms and testing the effects of candidate antimicrobial compounds on them. is there. The drip flow biofilm is generated on a coupon (substrate) in a groove of the drip flow reactor. Many different types of materials, such as ground 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 by dropping the medium into the chamber and then flushed to the length of the coupon at a 10 ° C. ramp.

  The biofilm is grown in a drip flow bioreactor and used alone or in combination with an antibiotic compound and / or with an antibiotic compound alone or in combination with another antimicrobial agent, including a BT compound. Or to 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 tilted polystyrene coupon in a covered chamber. An exemplary medium is a 5% v / v adult donor bovine serum (pH 6.8) that mimics high serum protein and very low iron concentration conditions 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 dropped (5 ml / h) onto four coupons contained in four separate parallel chambers, each 10 cm × 1.9 cm and 1.9 cm deep. The reactor divided into chambers was assembled with polysulfone plastic. Each of the chambers is fitted with an individual removable plastic lid that can be closed tightly. The biofilm reactor is placed in a 37 ° C. incubator and warmed by passing the bacterial cell culture through an aluminum heating bath maintained in the incubator. This method replicates the antibiotic-resistant phenotype observed within a particular biofilm and mimics the environment of low fluid shear and the closeness to the air interface characteristics of chronic wounds, while mimicking nutrients. Provides continuous replenishment and is compatible with a number of analytical methods to characterize and monitor the efficacy of the candidate antimicrobial regimen introduced. Drip flow reactors have been successfully used to culture a wide variety of pure and mixed species biofilms. Biofilms are typically grown for 2-5 days before application of antimicrobial agents.

  To determine the effect of an anti-biofilm agent on a biofilm grown in a drip flow reactor, a fluid stream passing over the biofilm is treated with a desired treatment formulation (eg, one or more BT compounds and / or Or 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 the biofilm is scraped into a beaker containing 10 ml of buffer. The sample is briefly processed (typically 30 seconds to 1 minute) with a tissue homogenizer to disperse the bacterial aggregates. The suspension is serially diluted and viable microorganisms are enumerated according to standard microbiological methodology.

Example 4
Inhibition of Scar Repair of Keratinocytes by Wound Biofilm: Biofilm Suppression by BT Compounds This example modifies an established in vitro keratinocyte scratch model of wound healing to biofilm-related wound pathology And relevance to wound healing, specifically reaching models with relevance to acute or chronic wounds or wound-containing biofilms as described herein. The keratinocyte scratch model for the effects of chronic wound biofilms allows the culture of mammalian (eg, human) keratinocytes and bacterial biofilm populations to proceed in separate chambers in fluid contact with each other, It allows the assessment of the effect of conditions affecting the effect of soluble components assimilated by biofilms on keratinocyte wound healing events.

  Neonatal human foreskin cells are cultured as monolayers in a treated plastic dish, in which controlled "wounds" or scratches are formed by mechanical means (eg, sterile scalpels, razors, Monophasic physical destruction, such as scratching the essentially linear cell-free zone between the monolayer regions using a suitable tool such as a cell scraper, tweezers, or other instrument). The in vitro keratinocyte monolayer model system is known to undergo structural and functional processes of cells in response to wound events in a manner that mimics wound healing in vivo. According to the embodiments disclosed herein, the effects of the presence of bacterial biofilms on such processes, e.g., on the healing time of a scratch, were observed and selected in these and related embodiments. The effect of the presence of the candidate antimicrobial (eg, antimicrobial and anti-biofilm) treatment is also evaluated.

  Wounded keratinocyte monolayers cultured in the presence of biofilms were examined by morphological, biochemical, molecular genetic, cytophysiological and other parameters, and the introduction of BT compounds Determine whether to alter the damaging effects of the film (eg, increase or decrease in a statistically significant manner relative to the appropriate control). To assess the effect of such a treatment on the effect of such treatment 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 with the BT compound Expose to combination.

  In an exemplary embodiment, a membrane (eg, such as a TransWell membrane insert) held in the previous tissue culture well and in fluid communication with a keratinocyte monolayer that is scratched to initiate the wound healing process. Above, the day 3 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 was 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 were cultured from neonatal foreskin as previously described (Fleckman et al., 1997 J Invest. Dermatol. 109: 36; Piepkorn et al., 1987 J Invest. Dermatol. 88: 215-219). Develop as a confluent monolayer on glass coverslips. The keratinocyte monolayer can then be scratched to create an even width “wound” and then the cell repair process can be monitored (eg, Tao et al., 2007 PLoS ONE 2: e697; But et al., 2007 Eur. J Cell Biol. 86: 747; Phan et al., 2000 Ann. Acad. Med. Singapore 29:27). The chamber is then assembled using aseptic technique, placing the artificial wound in the bottom of a sterile double-sided chamber. Keratinocyte growth medium with or without antibiotics and / or bismuth-thiol (EpiLife) is filled on both sides of the chamber. An inoculated system is used as a control.

  The bacteria isolated from the wound are inoculated into the system and incubated for 2 hours under static conditions to allow the bacteria to adhere to the surface in the upper chamber. Following the attachment period, a liquid medium flow is started in the upper chamber to remove unattached cells. Thereafter, the flow of medium is continued at a rate that minimizes the growth of platonic cells in the upper chamber by washing away unattached cells. After an incubation time in the range of 6-48 hours, the system (keratinocyte monolayer on coverslips and bacterial biofilm on membrane substrate) is disassembled, and coverslips are removed and analyzed. In a related embodiment, the chamber is assembled after the mature biofilm is grown in the upper chamber. In other related embodiments, at least one indicator of wound healing is altered, such as, for example, the time elapsed between when wound repair occurs or other signs of wound repair (eg, a statistical To identify agents or combinations of agents that increase or decrease in a manner that is significant to a candidate agent, such as a BT compound, or a potential synergistic BT compound + antibiotic combination for keratinocyte scratch repair For example, BT compounds provided herein, such as BT provided in particulate form, and amikacin, ampicillin, cefazolin, cefepime, chloramphenicol, ciprofloxacin, clindamycin (or other lincosamide antibiotics) ), Daptomycin (Cubicin®), doxycycline, gatifloxacin Biofilm and scraping to determine the effect of gentamicin, imipenim, levofloxacin, linezolid (Zyvox®), minocycline, nafcillin, paromomycin, rifampin, one or more of 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; But et al., 2007 Eur. J Cell Biol. 86: 747; Phan et al., 2000 Ann. Acad. Med. Singapore 29:27).

Example 5
Inhibition of Scar Repair of Keratinocytes by Wound Biofilm Isolated human keratinocytes were cultured on glass coverslips to create scratches according to the methodology described in Example 4 above. Wounded cultures were maintained on membrane supports in fluid communication with keratinocyte cultures, under culture conditions, alone or in the presence of co-cultured biofilms. The time interval of the wound closure during which the cell growth and / or migration of the keratinocytes re-established the keratinocyte monolayer across the scratch zone was measured. FIG. 3 shows the effect of the presence (but not in direct contact) of the biofilm in fluid communication on the healing time of the scratched keratinocyte monolayer.

  Thus, in certain embodiments, culturing a scratched cell (eg, keratinocyte or fibroblast) monolayer in the presence of a bacterial biofilm with or without a candidate anti-biofilm agent. And evaluating an indicator of healing of the scratched cell monolayer in the absence and presence of the candidate anti-biofilm agent, wherein the at least one indicator of healing is promoted. Drugs (eg, a BT compound such as a substantially monodispersed BT microparticle suspension described herein) alone or with an antibiotic such as amikacin, ampicillin, cefazolin, cefepime, chloramphenicol, ciprof Loxacin, clindamycin, daptomycin (Cubicin®), doxycycline, gatifloxacin, gentamicin Imipenim, levofloxacin, linezolid (Zyvox®), minocycline, nafcillin, paromomycin, rifampin, sulfamethoxazole, sulfamethoxazole, tobramycin and vancomycin) in an acute or chronic wound Or a method of identifying an agent for treating a chronic wound, identified as a suitable agent for treating a wound comprising a biofilm.

Example 6
Synergistic Bismuth-thiol (BT) Antibiotic Combination This example demonstrates the combination of one or more bismuth-thiol compounds with one or more antibiotics against various strains and strains, such as multiple antibiotic resistant bacteria. 2 shows examples of the demonstrated synergistic effects of the combinations.

  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) by broth dilution in 96-well tissue culture plates (Nalge Nunc International, Denmark).

  Briefly, an overnight bacterial culture was used to prepare a 0.5 McFarland standard suspension, which was then further supplemented with a cation-conditioned Mueller-Hinton broth medium (BBL, Cockeysville, MD, USA). Dilute 50 (~ 2 x 106 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 the manufacturer's recommendations. The minimum inhibitory concentration (MIC) was expressed as the lowest drug concentration that inhibited 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 the initial viability by 99.9% at 24 hours after inoculation.

  The activity of the antimicrobial combination was evaluated using a checkerboard method. The Partial Inhibitory Concentration Index (FICI) and the Partial Bactericidal Concentration Index (FBCI) were determined using Eliopoulos et al. Calculated according to Elliopoulos and Meollering, (1996) Antimicrobial combinations. Synergy was defined as a FICI or FBCI index of 0.5 or less, no interaction above 0.5 and no more than 4 and no more than 4 as antagonistic (Odds, FC (2003) Synergy, antagonism). , And what the chequerboard puts between them. Journal of Antimicrobial Chemotherapy 52: 1). As before, a four-fold decrease in antibiotic concentration was also defined as synergistic.

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 to 17.

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

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

BE = 0.2 μg / ml BisEDT; strain S2446-3 was obtained from Clinical Microbiology Laboratory of Winthrop-University Hospital, Mineola, NY. Antibodies were obtained from Sigma.

GM = gentamicin; strain S2400-1 was obtained from Clinical Microbiology Laboratory of Winthrop-University Hospital, Mineola, NY. Gentamicin was obtained from Pharmacy Department of Winthrop. Bold letters indicate synergy.

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

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

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

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

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

DOX = doxycycline; BE = BisEDT at 0.3 μg / ml; strain was obtained from Dr. of Department of Bacteriology, The University of Bristol, Bristol, UK. Obtained from the laboratory of I Chopra. Antibiotics were obtained from the Pharmacy Department of Winthrop-University Hospital, Mineola, NY.

Agr = aminoglycoside resistance; NN = tobramycin; PA = Pseudomonas aeruginosa; BE = BisEDT, 0.3 μg / ml; the strain is the Department of Microbiology and Immunology, Queens University, Canada, Oreg. K. Obtained from Poole's lab. Tobramycin was obtained from Pharmacy Department of Winthrop-University Hospital, Mineola, NY.

NN = Tobramycin; BE = BisEDT, 0.4 μg / ml; strain was obtained from Dr. of Department of Pediatrics and Communicable Disease, University of Michigan, Ann Arbor, MI. J. J. From LiPuma's lab; 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; strain was obtained from Dr. of Department of Pediatrics and Communicable Disease, University of Michigan, Ann Arbor, MI. J. J. From LiPuma's lab; 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; strain is Department of Chemistry and Biochemistry, Laurentian University, Ontario, Ontario. A. Obtained from Omri's laboratory (strain M is mucus-like B. cepacia; PA = Pseudomonas aeruginosa; SA = Staphylococcus 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; strain is Department of Chemistry and Biochemistry, Laurentian University, Ontario, Ontario. A. Obtained from Omri's lab (strain M is mucus-like B. cepacia; PA = Pseudomonas aeruginosa; SA = Staphylococcus 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 letters indicate synergy. 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 on Gram-positive and Gram-negative bacteria, including antibiotic-resistant strains. In this example, the in vitro activity of BisEDT and the comparator was determined by the gram-positive and gram-positive responsible for skin and soft tissue infections. Several clinical isolates of negative bacteria were evaluated.

Materials and Methods The test compounds and test concentration ranges are as follows: BisEDT (Domenico et al 1997; Domenico et al., Antimicrob. Agents. Chemother. 45 (5): 1417-1421, and Example 1), 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 (USP, NJ, # 1337809), 16-0.015 μg / mL and 8-0.008 μg / mL; ciprofloxacin (US Pharmacopeia, # 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 test system was 2.5%.

Organisms Organisms were obtained from the following clinical laboratories: CHP, Clarian Health Partners, Indianapolis, Ind .; UCLA, University of California, Los Angeles, California, USA, USA, Canada, USA. Center, Public Health Research Institute Tubeculosis Center, New York, NY; ATCC, American Type Culture Collection, Manassas, Va; Mt. Sina, VA; , 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 swabbed out of the isolation plate and placed in suspension in a suitable broth containing cryoprotectant. The suspension was dispensed into cold storage vials and kept at -80 <0> 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 Coccus aureus; 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 sensitive Enterococcus.

  Isolates can be obtained from frozen vials in a suitable medium (Trypticase Soy Agar (Becton-Dickinson, Sparks, MD) for most organisms) or Trypticase Soy Agar for Streptococcus spp. + 5% sheep blood, Cleavarian blood). 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% lysed horse blood (Cleveland Scientific Lot # H13913) and adapted to the development of Streptococcus pyogenes and Streptococcus agalactia. 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, for testing with daptomycin, the media was supplemented with an additional 25 mg / L Ca2 +.

  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 , 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. Automated liquid handlers included Multidrop 384 (Labsystems, Helsinki, Finland), Biomek 2000 and Multimek 96 (Beckman Coulter, Fullerton CA). 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 appropriate row and column 1 in these plates. These became mother plates when preparing test plates (daughter plates). With Biomek 2000, the series transfer to the eleventh row of the mother plate was completed. The wells in row 12 contained no drug and were the growth control wells on the daughter plate. 185 μL of the appropriate test medium (described above) was added to the daughter plate using Multidrop 384. Daughter plates were prepared by transferring 5 μL of drug solution from each well of the mother plate to each corresponding well of each daughter plate in one step on a Multimek 96 instrument.

  A standard inoculum of each organism was prepared by the CLSI method (ISBN 1-56238-587-9, cited above). The suspension was prepared in MHB to equal the turbidity of a 0.5 McFarland standard. The suspension was diluted 1: 9 with the appropriate broth for the organism. The inoculum of each organism was dispensed into a vertically sterile reservoir (Beckman Coulter) and the plates were inoculated using a Biomek 2000. The daughter plate was allowed to rest on the working surface of the Biomek 2000, which was inverted so that the inoculation was 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 wells of the daughter plate thus finally contained 185 μL of broth, 5 μL of drug solution, and 10 μL of bacterial inoculum. The plates were stacked in triplicate, the top plate was capped, 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, uninoculated soluble control plates were observed for evidence of drug precipitation. The MIC was read and recorded as the lowest drug concentration that inhibited the visual growth of the organism.

Results All commercial drugs were soluble at all test concentrations in both media. BisEDT showed a slight precipitate at 32 μg / mL, but did not affect the MIC reading because the inhibitory concentration of all organisms tested was well below that concentration. On each assay day, the appropriate quality control strain was included in the MIC assay. The MIC values obtained from these strains were appropriately compared with the published quality control ranges for each drug (Clinical and Laboratory Standards Institute 56. −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. The MIC values obtained from these strains were appropriately compared with the published quality control ranges for each drug (Clinical and Laboratory Standards Institute, Performance Standards for Medical Institutional Medical Institutional Medical Institutional Medical Examination, Medical, Medical, Medical, Pharmaceutical, Medical, Pharmaceutical, and Medical Sciences). -653-0]). Of the 141 quality control strains with published quality control ranges, 140 (99.3%) were within the specified range. One exception was imipenem against Staphylococcus aureus 29213 strain, where one single run was one dilution lower than the published QC range (<0.008 μg / mL). All other quality control results in its implementation were within the specified quality control scope.

  BisEDT demonstrates strong activity against methicillin-sensitive Staphylococcus aureus (MSSA), methicillin-resistant Staphylococcus aureus (MRSA), and community-acquired MRSA (CA-MRSA), and was tested at 1 μg / mL or less. 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 activity equivalent to daptomycin. Imipenem was more potent than BisEDT against MSSA (MIC90 = 0.03 μg / mL). However, while MRSA and CAMRSA were resistant to imipenem, 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 outside of imipenem. BisEDT was the most active drug tested for MRSE.

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

  BisEDT was highly active against vancomycin-sensitive Enterococcus faecium (VSEfm) with a MIC90 value of 2 μg / mL, its activity was similar or similar to daptomycin, and one dilution higher than vancomycin . BisEDT and linezolid were the most active agents tested against vancomycin-resistant Enterococcus faecium (VREfm), each of which demonstrated a MIC90 value of 2 μg / mL. The activity of BisEDT against Streptococcus pyogenes (MIC90 value 0.5 μg / mL) was equivalent to vancomycin, larger than linezolid, and slightly lower than daptomycin and ceftazidime. The compound inhibited all strains tested below 0.5 μg / mL. In these tests, the strain at least susceptible to BisEDT was Streptococcus agalactia and the observed MIC90 value was 16 μg / mL. BisEDT was less active than all tested drugs except gentamicin.

  The activity of BisEDT and comparative substances against the included Gram-negative bacteria demonstrated the efficacy of BisEDT against Acinetobacter baumannii (MIC90 value of 2 μg / mL), making BisEDT the most active compound tested. The MIC was elevated in a significant number of the test isolates for the comparison agents, which resulted in MIC90 values that were out of range for these agents. BisEDT is a potent inhibitor of E. coli and inhibited all strains below 2 μg / mL (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 an MIC90 value of 8 μg / mL, equivalent to imipenem. The relatively high MIC90 values exhibited by imipenem, ceftazidime, ciprofloxacin, and gentamicin indicated that this was a group of organisms highly resistant to antibiotics. BisEDT was the most active compound tested against Pseudomonas 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 exhibits broad spectrum capabilities against multiple clinical isolates representing multiple bacterial species, including those commonly involved in acute and chronic skin and skin structure infections in humans. The activity of BisEDT and key comparators was evaluated against 723 clinical isolates of Gram-positive and Gram-negative bacteria. The BT compound demonstrated broad spectrum activity, and for many test organisms in this test, BisEDT was the most active compound tested for antimicrobial activity. BisEDT is available from MSSA, MRSA, CA-MRSA, MSSE, MRSE and S.E. It was most active against P. pyogenes, with MIC90 values below 0.5 μg / mL. Strong activity has been demonstrated in VSEfc, VREfc, VSEfm, VREfm, A. Baumannii, E. coli, and Pseudomonas aeruginosa were also demonstrated, with MIC90 values in the range of 1-4 μg / mL. If the MIC90 value is Pneumoniae (MIC90 = 8 μg / mL) and Agaractia (MIC90 = 16 μg / mL) was observed.

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

  A major complicating factor in treating infections 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. In addition, subMIC concentrations of BT reduced resistance to several important antibiotics.

  Providing a graphical representation of the antibiotic-resensitizing effects of subMIC bismuthethanedithiol (BisEDT) on Staphylococcus aureus MRSA (FIG. 4), gentamicin, cefazolin, cefepim, simipem, imepime Multiple classes of antibiotics, including methoxazole and levofloxacin, show potent antibiotic action. 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 the biofilm preventive concentration (BPC) and the minimum inhibitory concentration (MIC) were measured on a special biofilm medium (BHIG / X). Gentamicin and cefazolin MIC and BPC were reduced by subMIC BisEDT (BisEDT MIC, 0.2-0.4 μg / mL), but not below the breakpoint of sensitivity. SubMIC's BisEDT enhanced the sensitivity of MRSA to gatifloxacin and cefepime to near the breakpoint of sensitivity. These strains were already sensitive to vancomycin, but were even more pronounced in the presence of subMIC BisEDT. In general, MIC and BPC were reduced 2- to 5-fold by subMIC's BisEDT.

Twelve MRSA clinical isolates were grown in BHIG / X and exposed to serial dilutions of the antibiotic in the presence of 0-0.1 μg / mL BisEDT. MICs and BPCs calculated in μg / mL are means ± SD from at least 3 trials. The right column lists the 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 studies of cefepime resistant MRSA isolates are shown in Table 19. 0.1 μg / mL BisEDT significantly enhanced cefepime inhibitory activity in 11 of the 12 isolates. In this particular study, the data showed a synergy between BisEDT and cefepime (FIC <0.5), with many of the isolates at sensitive breakpoints.

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

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 MIC90 of nafcillin by more than 4-fold over MRSA (FIC, 0.74). BisEDT in combination with gentamicin reduced gentamicin MIC90 by more than 10-fold relative to 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 respect to gentamicin. The broth used in these tests was Trypticase Soy Broth (TSB) containing 2% glucose and showed similar results to those seen in Mueller-Hinton II broth supplemented with 1% sheep blood.

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

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

  A slight decrease in biofilm prophylactic concentration (BPC) was observed with minocycline, vancomycin, and cefazolin, while rifampicin and nafcillin still had no effect at 0.05 μg / mL BisEDT. At 0.1 μg / mL BisEDT, no biofilm was detected, regardless of the antibiotic used, indicating that no antagonism occurred. This BisEDT concentration was determined by Epidermidis MIC [Domenico et al. , 2003] (see FIG. 5).

  For stunting, seven of the eight antibiotics tested were S. Epidermidis was significantly enhanced in the presence of 0.1 μg / mL (0.5 μM) BisEDT (FIG. 6). The MIC variability was most pronounced for clindamycin and gentamicin, followed by vancomycin, cefazolin, minocycline, gatifloxacin and nafcillin, and rifampicin had no effect. These strains were resistant to the antibiotics (NC, CZ, GM, CM) 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's BisEDT. Gentamicin showed the greatest reduction in MBC (4- to 16-fold), followed by cefazolin (4- to 5-fold), vancomycin and naphcillin (3- to 4-fold), minocycline and gatifloxacin (2- to 3-fold). ) And the MBCs of clindamycin and rifampicin were still not significantly affected. Clindamycin is a bacteriostat and is described as having no bactericidal activity. Cefazolin resistance was suppressed for MBC [Domenico et al. , 2003]. These effects were additive.

  The efficacy of antimicrobial agents was also demonstrated in a rat graft infection model in vivo (Table 21). With a low level of BisEDT as low as 0.1 μg / mL, resistant S. Epidermidis biofilm prevention could be promoted.

As summarized in Table 21, grafts impregnated with 0.1 μg / mL BisEDT, 10 μg / mL RIP and 10 μg / mL rifampin, alone or in combination, were implanted subcutaneously in rats. A physiological solution (1 ml) containing 2 × 10 7 cfu / ml MS and MR strains was inoculated onto the surface of the implant using a tuberculin syringe. All implants were removed 7 days after implantation and sonicated in sterile saline for 5 minutes to remove attached bacteria. The quantification of surviving bacteria was obtained by incubating 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-fold by subMIC BisEDT (Table 22). In these trials, the MIC was more accurately defined as IC24.

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

Against tobramycin-resistant Burkholderia cepacia, 0.4 μg / mL BisEDT sensitized seven of the ten isolates to tobramycin (mean FIC; 0.48) and reduced MIC90 by 10-fold ( Table 23). Both MIC and MBC of tobramycin were significantly reduced by subMIC BisEDT to levels achievable for 50 clinical Burkholderia cepacia isolates [Veloria et al. , 2003]. Liposomal forms of BisEDT and tobramycin have demonstrated high synergism 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 by the addition of subMIC BisEDT (Table 24).

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

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 Escherichia coli became sensitive to tetracycline upon addition of subMIC BisEDT (Table 25). The combination was synergistic for the TET M and TET D strains (FIC ≦ 0.5) and showed an additive effect on the TET A and TET B strains.

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

References Domenico P, RO'Leary, BA Cunha. 1992. Differential effect of bismuth and salicylate compounds on antibiotic sensibility of Pseudomanas aeruginosa. Eur J Clin Microbiol Infect Dis 11: 170-175; Domenico P, D Parik, BA Cunha. 1994. Bismuth modulation of antibiotic activity against gastrointestinal bacterial pathogens. Med Microbiol Lett 3: 114-119; Domenico P, Kazzaz JA, Davis JM, Niederman MS. 2002. Subinhibitory bismuth ethanolithiol (BisEDT) sensitives 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; Domenico P, E Gurzenda, A Giacometti, O Cironi, R.G.A.R., G.A.R., G.R. 2004. BisEDT and RIP act in synergies to present graft infections by resistant staphylococci. Peptides 25: 2047-2053; Halwani M, Bromme S, Suntres ZE, Alipour M, Azghani AO, Kumar A, Omri A. 2008. Liposomal bismuth-ethanidithiol formation enhancement enhances antimicrobial activity of tobramycin. Intl J Pharmaceut 358: 278-84; Halwani M, Hebert S, Suntres ZE, Lafrenie RM, Azghani AO, Omri A. 2009. Bismuth-thiol incorporation enhancements biological activities of liposomal tobramycin against bacterial biofilm eumodose quantosumeduscologouse euduscos muscodusume sul grosco sul gross estroscos Int J Pharmaceut 373: 141-6; Veloila WG, Gurzenda EM, Domenico P, Davis JM, Kazzaz JA. 2003. Synergy of tobramycin and bismuth thiols against Burkholderia sepacia. 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
Microparticle BT-Enhancement and Synergistic Activity of Antibiotics This example demonstrates that microparticle bismuth thiol, BisEDT, has been shown to enhance antibiotic activity through potent and / or synergistic interactions with specific antibiotics against specific microbial target organisms. To promote. One-point data of each combination shown in Table 26 was prepared 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 multivolans; Abau, Acinetobacter baumanii; Mseg, Mycobacterium smegmatis

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

  Stock solutions of all test substances were prepared at 40 times the final desired concentration in the appropriate solvent. All test substances were solutions under these conditions. The final drug concentration in the FIC assay plate was set so that the MIC values of each drug for each test organism were batched, unless the strain was totally resistant to the test agent. The tested concentration ranges are 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 picked from these plates and cell suspensions were prepared in a suitable broth growth medium containing cryoprotectants. Thereafter, aliquots were frozen at -80 ° C. Frozen seeds of the organism to be tested in a given assay were thawed, streaked onto TSA plates for isolation, and incubated at 35 ° C. All organisms were examined on a Mueller-Hinton II Broth (Becton Dickinson, lot number 9044411). Broth was prepared at 1.05 times normal weight / volume to offset 5% volume of drug in the final test plate.

  The final inhibitory concentration (MIC) value was measured as before using the broth microdilution method for aerobic bacteria (Clinical and Laboratory Standards Institute (CLSI). Methods for Digestion Biotaxis Biostitial Biostitial Biotissues). Applied Standard-Eight Edition.CLSI document M07-A8 [ISBN 1-56238-689-1] .Clinical and Laboratory Standards Institute, 940 Westward Valuables. 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 dilutions and transfers were performed using an automated liquid handler (Multidrop 384, Labsystems, Helsinki, Finland; Biomek 2000 and Multimek 96, Beckman Coulter, Fullerton CA).

  Using Multidrop 384, the appropriate wells of a standard 96-well microdilution plate (Falcon 3918) were loaded with 150 μL of the appropriate solvent from rows 2 through 12. 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 a "mother plate" of the drug that provided serial dilutions for the drug combination plate. 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-fold dilution series. Mother plates of Bis-EDT (and analogs) 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 the same volume (using a multichannel pipette) to the drug combination plate This formed a "checkerboard" pattern. Row H and column 12 each contained serial dilutions of one of the drugs whose MIC was measured alone.

  180 μL of the test medium was added to the “daughter plate” using Multidrop 384. Thereafter, using Multimek 96, 10 μL of the drug solution was transferred in one step from each well of the drug combination mother plate to each corresponding well of the daughter plate. Finally, daughter plates were inoculated with the test organism. 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 sterile vertically separated reservoirs (Beckman Coulter) and the plates were inoculated using a Biomek 2000. The instrument delivered 10 μL of the standardized inoculum to each well to give a final cell concentration of approximately 5 × 10 5 colony forming units / mL in the daughter plate.

  The test format produced an 8 × 12 checkerboard for testing each compound alone (columns 12 and 8) and combining drug concentrations in various ratios. All organism plates were stacked in triplicate, covered with a top plate, 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 the ScienceWare plate viewer. On the prepared reading sheet, the MIC of drug 1 (line H), the MIC of drug 2 (column 12), and the wells with and without growth were entered.

  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). FICI for the checkerboard was calculated from each FIC by the formula: (FIC1 + FIC2 + ... FICn) / n, where n = number of wells per plate for calculating FIC. If a single drug resulted in an out-of-measurement MIC result, the next highest concentration was used as the MIC value in the FIC calculation.

All of the particulate Bis-EDT, the four particulate 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 the rat femur critical defect The current standard of care for open fractures is irrigation, debridement and antibiotics, which do not result in infection of bacterial mass in the wound It is intended to be reduced to a degree. Despite these procedures, infection complicates up to 75% of severe open tibial fractures due to war. Interestingly, even though the initial infection is often caused by Gram-negative bacteria, the late infections involved in healing problems and amputation are due to Gram-positive bacterial infections, mostly Staphylococcus species (Johnson 2007).

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

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

  Compound (CPD) CPD-8-2 (bismuth-pyrithione / butanedithiol; Table 1) and CPD-11 (bismuth pyrithione / ethanedithiol; Table 1), despite having a different spectral activity than Bis-EDT , Two BIS-Bis analogs that have shown potency against 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 polymethyl methacrylate (PMMA) cement bead vehicle. When used, it had an inhibitory effect on S. aureus strains in vitro. Three formulations of particulate BT were prepared in the clinically useful hydrogel forms described herein. These BTs were tested suspended in the gel at a concentration of 5 mg / ml-1 which was found to be the appropriate concentration for gel delivery. The gel formulation conformed to the wound contour and did not need to be removed after application.

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

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

(B) BT and systemic antibiotics (ABx)
Six hours after inoculation of the Staphylococcus aureus, the wound was debrided, ligated with saline and 1 ml of the added BT gel was inserted into the defect. The antibiotic used was cefazolin at a dose 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 just before debridement. Past data suggested that this dose could reduce bacterial levels from about 106 to about 104, and thus measure the relative effects of different BTs.

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

procedure:
The procedure of the in vivo rat injury model was 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). 2007 J. Orthop. Trauma 21: 693). The rats were anesthetized and prepared for surgery. The anterior lateral aspect of the femoral shaft was exposed by incision of 3 cm. 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 threaded through a 0.9 mm diameter twisted Kirschner wire. The substrates of these plates were shaped to fit the contour of the femoral shaft. Pilot holes were cut through both femoral cortical bones using the plate as a template, and a twisted Kirschner wire was inserted through the plate and femur. Indentations at 6 mm intervals on the plate provided guidance for bone removal. In an attempt to prevent thermal damage, a defect was created using a small oscillating saw while the tissue was cooled by continuous irrigation.

  Each group of 10 animals was inoculated with 1 × 10 5 CFU of S. aureus and treated with BT alone or in combination with antibiotics 6 hours post-inoculation as described above. 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) ).

  Animals were euthanized 14 days after surgery and bones and devices were sent for microbial analysis, the results of which are shown in FIG.

  Based on power analysis, 10 animals per group will provide 80% power to detect a 25% difference between the treated 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, the antibiotic activity of cefazolin is enhanced as compared to cefazolin or any Bis compound alone, and the yellow color of damaged bone Reduced staphylococcal infections. Cefazolin in combination with MB-11 and MB-8-2 showed higher antibiotic activity compared to cefazolin alone, indicating that it reduced the S. aureus infection detected on the device. Bis-EDT was not expected to affect cefazolin activity in this capacity.
Example 12
Activity of bismuth-containing compounds on 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 a bismuth-containing compound. The MIC values of three bismuth-containing compounds for three different marine bacteria, namely bismuth dimercaprol (BisBAL), bismuth dimercaptotoluene (BisTOL) and bismuthethanedithiol (BisEDT), are routinely performed by those skilled in the art. Was determined using the method described. The data is shown in the table below.

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

Compounds BisBAL and BisTOL were included in the assay to determine the inhibitory activity of each compound on barnacle larval settlement behavior. The method was performed according to techniques practiced in the art. BisBAL had an EC50 of 1.6 ppm (the concentration at which 50% settle inhibition occurs) and BisTOL had an EC50 of 15.4 ppm. In another experiment, BisEDT was directly dissolved in natural seawater or first dissolved in DMSO and then diluted into natural seawater. EC50 measurements were not statistically different. BisEDT had an EC50 of 1.5 ppm when directly dissolved in seawater and had an EC50 of 2.1 ppm when first dissolved in DMSO. The commercial biocide SEANINE 211 had an EC50 of 0.5 ppm.
Example 14
Effects of Bismuth-Containing Compounds on Algal Settlement

The action of three bismuth-containing compounds on the algae settlement, namely, bismuth dimercaprol (BisBAL), bismuth dimercaptotoluene (BisTOL) and bismuthethanedithiol (BisEDT) Determined 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 the germination of about 50% of the algal spore population and 10 μg / ml inhibited the germination of about 75% of the algal spores. Up to 10 micrograms / ml of BisBAL and BisTOL had no inhibitory effect on the germination of spores of this particular phase species.
Example 15
Effects of Bismuth-Containing Compounds on Algal Settlement

  The effect of three bismuth-containing compounds on the growth of marine diatoms, namely bismuth dimercaprol (BisBAL), bismuth dimercaptotoluene (BisTOL) and bismuthethanedithiol (BisEDT), is a technique implemented in the art. Determined according to Increasing the concentrations of the three compounds (0.001, 0.01, 0.1, 1.0 and 10.0 μg / ml) suppressed marine diatom settlement (diatoms / pits). . Each compound showed inhibitory activity at 0.1 μg / ml; BisEDT was the most active, showing almost 100% inhibition. BisTOL and BisBAL each showed about 30% marine diatom settlement at 0.1 μg / ml.

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  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 sheets are incorporated herein by reference in their entirety. Will be incorporated into the book. Aspects of the embodiments can be refined 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 shall not be construed as limiting the claims to the particular embodiments disclosed herein and in such claims. It is intended that the following claims be interpreted as covering all possible embodiments as well as the full scope of equivalents to which it is entitled.

Claims (20)

Bismuth thiol (BT) compositions in the manufacture of a medicament for the treatment of open, chronic or acute wounds at risk of having or having a bacterial biofilm infection The use of
The use wherein the BT composition comprises a monodisperse suspension of solid microparticles of bismuth-ethanedithiol (BisEDT) having a volume average diameter of 0.4 μm to 5 μm.   The use according to claim 1, wherein the open, chronic, or acute wound is an open wound.   3. Use according to claim 2, wherein the open wound comprises an open fracture.   The use according to claim 1, wherein the open wound, chronic wound, or acute wound is a chronic wound.   5. Use according to claim 4, wherein the chronic wound comprises a skin infection or a soft tissue infection or both.   The use according to claim 1, wherein the open wound, chronic wound, or acute wound is an acute wound.   7. Use according to any of the preceding claims, wherein the open, chronic or acute wound comprises an epithelial tissue surface.   8. Use according to any one of the preceding claims, wherein the open, chronic or acute wound comprises (i) a natural surface, or (ii) a natural and artificial surface.   4. Use according to any one of the preceding claims, wherein the medicament is a bone cement.   The use according to claim 9, wherein the composition comprises a hydrogel.   The use according to claim 10, wherein the hydrogel contains polymethyl methacrylate (PMMA). The use according to claim 1, wherein the bacterial biofilm infection comprises at least one of the following items (i) to (v):
(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; and (v) Staphylococcus aureus (Staphylococcus aureus), MRSA (Methicillin-resistant Staphylococcus aureus), Staphylococcus epidermidis, MRSE (Methicillin-resistant S. epidermidis). , Mycobacterium tuberculosis, 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 faecalis, methicillin-sensitive Enterococcus faecalis, Enterobacter cloacae, Salmonella typhimurium, Proteus bulgaris, El Near Enterocolitica, Vibrio cholere, Shigella flexneri, Vancomycin-resistant Enterococcus (VRE), Burkholderia cepacia, Francisella tularensis, Bacillus anthracis, Yersinia pestis, Pseudomonas aeruginosa, Streptococcus pnemoninus A bacterial pathogen selected from resistant Streptococcus pneumoniae, Burkholderia cepacia, Burkholderia multiborans, Mycobacterium smegmatis and Acinetobacter baumanny. Use of a bismuth thiol (BT) composition in the manufacture of a medicament for protecting a product from bacterial, fungal, or viral pathogens,
The medicament is contacted with or incorporated into the product in an effective amount of the BT composition in a sufficient condition or time for one or more of the following (i) to (iv): Used for
(I) prevention of infection of the product by bacterial, fungal or viral pathogens;
(Ii) inhibiting the cell viability or cell proliferation of substantially all planktonic cells of a bacterial, fungal or viral pathogen;
(Iii) inhibition of biofilm formation by a bacterial, fungal or viral pathogen; and (iv) inhibition of biofilm viability or biofilm growth of substantially all biofilm-type cells of the bacterial, fungal or viral pathogen;
Here, the BT composition comprises a monodispersed suspension of solid fine particles of bismuth-ethanedithiol (BisEDT) having a volume average diameter of 0.4 μm to 5 μm, and bringing the drug into contact with the surface of the product. Or incorporating the medicament into the product is performed in vitro.
use.   14. Use according to claim 13, characterized in that the surface of the product is (i) present on a medical device or implant or (ii) present on a dental device or implant. .   15. The use according to claim 14, wherein the medical device or the medical implant is an artificial bone, an artificial joint, a catheter, a stent, a feeding tube, or a gastrostomy tube.   15. Use according to claim 13 or claim 14, wherein the product comprises a cement surface, a concrete surface, a rubber surface, a silicone surface, a plastic surface, a paint surface, or a coated surface. Said product is protected from bacterial pathogens;
The product comprises an artificial bone or an artificial joint, and the BT composition contains fine particles of the BT compound in bone cement.
14. Use according to claim 13.   18. Use according to claim 17, wherein the composition comprises a hydrogel.   19. Use according to claim 18, wherein the hydrogel contains polymethyl methacrylate. 20. Use according to any one of the preceding claims, characterized in that the microparticles are not micronized, milled or undergo supercritical fluid processing.

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