MXPA05013038A

MXPA05013038A - Antiseptic compositions, methods and systems

Info

Publication number: MXPA05013038A
Application number: MXPA/A/2005/013038A
Authority: MX
Inventors: Kite Peter; Hatton David
Original assignee: Aseptica Inc
Priority date: 2003-06-04
Filing date: 2005-12-02
Publication date: 2006-10-17

Abstract

Antiseptic compositions comprising at least one salt of EDTA are disclosed. These compositions have broad spectrum antimicrobial and antifungal activity together with anticoagulant properties. The antiseptic compositions also have demonstrated activity in penetrating and breaking down microbial slime, or biofilms. They are safe for human and medical uses, and may be used to prevent infection, or to reduce the proliferation of and/or eliminate existing or established infections.

Description

ANTISEPTIC COMPOSITIONS, METHODS AND SYSTEMS FIELD OF THE INVENTION The present invention relates to antiseptic compositions, methods and systems for use in various medical applications, as well as disinfection applications in general, including industrial and environmental disinfection applications. The compositions of the present invention possess antimicrobial, anti-fungal, anti-viral and anti-ammic properties and can also serve as anti-coagulants. The specific salts and compositions of ethylenediamine tetraacetic acid (EDTA) (C10H12N2Na4O8) are used at specific concentrations and pH levels. Exemplary applications include inhibition, reduction or elimination of the presence of microbial and / or fungal organisms on surfaces, in solutions, or in complex forms, such as in biofilms. Exemplary methods include providing an antiseptic coating on object surfaces to reduce the incidence of infection, and contacting objects and / or surfaces by water jetting, soaking and / or rinsing with an antiseptic solution to inhibit proliferation, reduce or eliminate microbial populations. BACKGROUND OF THE INVENTION Infections are a significant problem in ßf. 168652 many fields where sanitary conditions are important, such as in health care. Problematic infections can arise from bacterial, fungal, amoebic, protozoal and / or viral organisms. The challenges are as much in the prevention of infection as in the reduction or elimination of the infection once it is established. Infected environments may include surfaces of objects, fluids and fluid conduits, and / or humans or animals. Alcohol solutions and wipes with isopropyl alcohol are commonly used to disinfect surfaces and have been shown to have antibacterial activity. The most effective anti-microbial inhibitory effect is seen with 70% isopropanol solutions. Alcohol solutions at this concentration are quite expensive and evaporate quickly, which substantially decreases their efficiency and increases their cost. In addition, although isopropanol solutions can be used on surfaces, including human skin, and in a variety of medical applications, alcohol solutions of this concentration can only be administered topically to humans for medical purposes. In the field of health care, infections of various types and causes are common and often result in longer hospital stays, leading to higher hospital costs. Even worse, over 90,000 patient deaths annually are attributed to nosocomial infections - that is, infections acquired in a hospital or other healthcare setting. Nosocomial infection surveillance has become an integral part of hospital practice. The studies conducted more than 20 years ago by the Centers for Disease Control and Prevention (CCE) document the effectiveness of these surveillance activities in reducing the occurrence of nosocomial infection. Despite the attention given to nosocomial infection problems, however, the proportions of infection have not been dramatically reduced, and nosocomial infections remain with a substantial risk and a substantial health interest. A problematic source of infections in the medical and veterinary fields is found in catheters, and particularly in resident catheters. Catheters have become essential in the management of intensive care patients, yet the inside of a catheter is often the main source of infection. Catheters are used to deliver fluids, blood products, drugs, and nutrients, as well as for hemodialysis, hemofiltration, peritoneal dialysis, blood sample collection, patient condition monitoring, etc. Transcutaneous catheters frequently become infected through penetration of the skin of the catheter. It has been found that seventy percent (70%) of all nosocomial bloodstream infections occur in patients with central venous catheters. Daouicher et al. New Engl. J. Med. 340: 1-8 (1999). In particular, during some procedures, a catheter must be implanted, and remain implanted, in a patient by a? relatively long period of time, for example, for thirty days. Intravenous (IV) therapy catheters and urinary catheters typically remain implanted for a substantial period of time. As a result of trauma to the areas of insertion, and pain to patients, such catheters can not be removed and implanted frequently. Bacteria that develop the catheter have been a primary source of urinary tract infections. Patients who receive a peripherally inserted central catheter during pregnancy have also been found to be at significant risk for infectious complications. Ogura et al. Am. J. Obstet. Gynecol. 188 (5): 1223-5 (2003). In addition, infection of the central venous catheter, which results in catheter-related sepsis, has been cited as the most common complication during parenteral household nutrition. Reimund et al. Clinical Nutrition, 21 (l): 33-38 (2002). Due to the risk of infections, catheterization can be limited to incidence when the procedure is absolutely necessary. This seriously compromises the patient's health. After the most prescribed medical access procedures involving a catheter, the catheter is flushed with saline and then filled with a liquid, such as saline solution or heparin, to prevent blood from clotting inside the catheter. catheter, inhibit the patient's blood from getting stuck in the catheter, and prevent gas from entering the catheter. The liquid used to flush the catheter is referred to as a "rinse and rinse," and the liquid used to fill the catheter after jet washing or during periods of non-use is referred to as a "rinse" solution. Traditionally, catheters have been rinsed with normal saline or heparin solutions, which provide anticoagulant activity. Heparin and saline are sometimes used in combination. Normal saline is usually used to rinse short-term peripheral intravenous catheters, but has no anticoagulant or antimicrobial activity. Heparin solutions are generally used to flush vascular catheters. Heparin has anticoagulant activity but does not work as an antimicrobial and does not prevent or improve infections. There are also strong indications that heparin in rinse solutions may contribute to heparin-induced thrombocytopenia, a serious bleeding complication that occurs in a subset of patients receiving heparin injections. Catheter rinsing solutions comprising Taurolidine, citric acid and sodium citrate have been proposed. A recent publication (Kidney International, Sept. 2002) describes the use of a 70% alcohol solution as a rinsing solution for a subcutaneous catheter orifice. The use of alcohol as a rinse solution is questionable, since it is not an anticoagulant, and since there may be risks associated with this solution entering the bloodstream. The inventors are also unaware of any evidence indicating that a 70% alcohol solution has some biofilm eradication activity. A recommendation and opinion that emerges from the Center for Infectious Diseases (CIS) is to treat existing catheter infections systematically with either a specific or broad-range antibiotic. The use of an antibiotic in a rinse solution to prevent infection is not recommended. The use of antibiotics to treat existing catheter infections has certain risks, including: (1) the risk of developing antibiotic-resistant strains; (2) the inability of the antibiotic to kill sessile, or deep-layer biofilm, bacteria, which may require the use of antibiotics at toxic concentrations; and (3) the high cost of prolonged antibiotic therapy. Catheters coated with an antiseptic or antibiotic material are available. However, these coated catheters can only provide limited protection for a relatively short period of time. In general, free floating organisms can be vulnerable to antibiotics. However, bacteria and fungi can become impermeable to antibiotics by adding to surfaces and producing a viscous protective substance, often referred to as extracellular polymeric substance (SPE) to form a biofilm. When microbes proliferate "more than fifty over or under genetic regulations can occur, resulting in the formation of a resistant microbial biofilm plus antibiotic. Two thirds of bacterial infections that doctors find have been attributed to biofilms. Netting, Science News, 160: 28 (2001). Biofilm formation is a genetically controlled process in the life cycle of bacteria that produces numerous changes in the cellular physiology of the organism, often including increased antibiotic resistance (up to 100 to 1000 times), when compared to growth under planktonic conditions ( free floating). When organisms grow, problems with the accumulation and decrease of recreation of nutrition activator organisms to find new locations and resources. The recently spread organisms quickly revert back to their original free floating phase and once again they are vulnerable to antibiotics. However, the free floating organism can enter the bloodstream of the patient, creating infections in the bloodstream which produce clinical signs, for example, fever, and more serious symptoms related to infection. Sesame biofilm ponds can detach and attach to tissue surfaces, such as heart valves, causing biofilm proliferation and serious problems, such as endocarditis. In industrial facilities, biofilm formation is very common and is generally referred to as biofouling. For example, the growth of biofilm in mechanical structures, such as filtration devices, is a primary cause of biological contamination of drinking water distribution systems. Biofilm formation in industrial facilities can lead to material degradation, product contamination, mechanical blockage and heat transfer impedance in processing systems. Biofilm formation and the resulting contamination is also a common problem in the preparation of food and processing facilities.

For additional complicated issues, conventional sensitivity tests measure only the antibiotic sensitivity of free floating organisms, rather than organisms in a biofilm state. As a result, a dose of antibiotics is administered to the patient, such as through a catheter, in amounts that rarely have the desired effect on biofilm-phase organisms that may reside in the catheter. Biofilm organisms can continue to spread more planktonic organisms or they can go dormant and proliferate later as an apparent recurrent infection. To eradicate biofilm organisms through the use of antibiotics, a laboratory must determine the concentration of antibiotic required to kill the specific biofilm phase of the organism. The highly specialized team is required to provide the minimum concentration for biofilm eradication. In addition, current diagnostic protocols are time consuming, and results are often not available for many days, for example five days. This period of time clearly does not allow the rapid treatment of infections. This delay, and well-justified infection concern, can result in the overuse of broad-spectrum antibiotics and unnecessary unnecessary catheter removal and replacement procedures. Excessive use of broad spectrum antibiotics can result in the development of antibiotic resistant bacterial strains which can not be effectively treated. The replacement and removal of unnecessary catheter is painful, costly and can result in trauma and tissue damage at the catheter insertion site. Antibiotic resistance of biofilms, coupled with complications of antibiotic use such as the risk of the development of antibiotic resistant strains, has made antibiotic treatment an unattractive option. As a result, the use of antibiotic is limited to symptomatic infections and prophylactic antibiotics are not typically employed to prevent contamination. Because the biofilm can act as a selective phenotypic resistance barrier to most antibiotics, the catheter must often be removed to eradicate a catheter-related infection. The removal and replacement of the catheter takes time, disturbs the patient, and complicates the medical procedure. Therefore, there is a need for convenient and effective methods to kill organisms, and especially those living within catheters, without the need to remove the catheter from the body. In addition to bacterial and fungal infections, amyloid infections can be very serious and painful, as well as potentially life threatening. It has been found that various species of Acanthameoba, for example, infect humans. Acanthamoeba are found in the world on earth and dust, and in freshwater sources as well as in brackish water and seawater. They are often found in heating, venting, and air conditioning units, in humidifiers, dialysis units, and in contact lens paraphernalia. Acanthamoeba infections, in addition to microbial and fungal infections, may also be common in connection with other medical and dental devices, including toothbrushes, dentures and other dental applications, and the like. Acanthamoeba infections often result from incorrect storage, handling and disinfection of contact lenses and other medical devices that come in contact with the human body, where they can enter the skin through a cut, wound, the nostrils, the eyes, and the like. There are numerous different types of microbes that present problematic infections, including varieties of bacteria and fungi. However, the present methods of infection elimination generally employ solutions that are effective against a limited number of different microbes. Root efc al. (Antimicrobial Agents and Chemo therapy, 32: 1627-1633 (1988)) describes the in vitro use of disodium EDTA against an isolate of pathogen Staphylococcus epidermidis associated with catheter, particular. EDTA has traditionally been useful as a metal chelator and has been used, in combination with other active compounds, for a variety of purposes. EDTA is frequently used, in low concentrations, as an anticoagulant in vi tro for testing and collection of blood specimens and as an antioxidant synergist, and is also added to solutions, for example, as a chelator, a stabilizer, or a preservative for pharmaceutical preparations. EDTA can exist in a variety of forms, some of which are sodium salt forms, such as disodium, trisodium and tetrasodium salts, and others of which are metal chelates such as iron, copper, magnesium, etc. Certain forms of EDTA have been used, in conjunction with other substances as an adjuvant, in compositions for treating infected catheters. When used in a clinical setting, or in a composition used with humans or animals, the solutions generally conform to a physiological, or neutral pH range. A combination of an alcohol with an additive, such as a salt-free form of EDTA, is described in PCT publication WO 02/05188. PCT publication WO 00/72906 A1 discloses a lyophilized mixture of an antimicrobial agent, for example an antibiotic, and a second agent, which may be a sodium-free salt form of EDTA, as a chelating agent for catheter jet wash . In U.S. Patent No. 5,688,516, compositions having an anticoagulant, a chelating agent, such as EDTA, and an antimicrobial agent, such as Minocycline, are described for coating medical devices and inhibiting catheter infection. In particular described examples, a disodium form of EDTA is delivered to a physiological pH of 7.4 and used in the composition. PCT publication WO 99/09997 describes the treatment of fungal infection with a combination of an antifungal agent and a chelator, such as EDTA. Other areas in which infections present a problem include medical devices and materials used in connection with the eyes, such as contact lenses, scleral loops, suture materials, intraocular lenses, and the like. In particular, there has been an emphasis on the development of methods for disinfecting ocular prostheses, for example contact lenses. Bacterial biofilms can participate in ocular infections and allow bacteria to persist on abiotic surfaces that come in contact with, or are implemented with, the eyes. Biofilms can also form on the biotic surfaces of the eye. Zegans et al. , DNA Cell Biol. , 21: 415-20 (2002). A severe form of keratitis can also be initiated by a protozoan amoeba which can contaminate lens disinfectant fluids. An ophthalmic formulation of tetrasodium EDTA and alkali salts, buffered at a pH of 6-8, for disinfecting contact lenses is described in U.S. Patent No. 5,300,296. U.S. Patent No. 5,998,488 describes an ophthalmic composition of EDTA and other substances, such as cyclodextrin and boric acid. In the dental field, articles to be placed in a mouth, such as dental tools, and dental and orthodontic devices such as retainers, bridges, dentures, and the like, need to be maintained in a sterile condition, particularly during storage and prior to the placement in the mouth. Otherwise, the infection can be transmitted to the bloodstream and become serious. U.S. Patent No. 6,077,501 discloses the use of EDTA in a denture cleansing composition with other active components. The water supply is also prone to microbial infections and other types of infections. Water storage devices, as well as water supply and extraction ducts, frequently become infected. The internal surfaces of the pipe that carries fluid in medical and dental services present an environment that is suitable for microbial growth and infection, and the adherence of microbes and formation of highly protective biofilm layers is often problematic in storage and supply devices. fluid . There is a need for improved methods and compositions to prevent and destroy infections in a variety of environments. Such antiseptic solutions should have a wide range of antimicrobial properties. In particular, solutions must be able to penetrate biofilms to eradicate organisms in biofilms. The methods and solutions should be sufficiently safe to be used as a preventive measure as well as in the treatment of existing infections.

BRIEF DESCRIPTION OF THE INVENTION The present invention provides antiseptic solutions comprising, consisting essentially of, or consisting of, one or more salts of EDTA at a pH that is greater than the physiological pH and at a prescribed concentration. The inventors have discovered that certain EDTA compositions possess potent antiseptic activities and function as broad spectrum anti-microbial agents, as well as function as fungicidal agents against many strains of pathogenic yeast. The EDTA compositions of the present invention are also highly effective in the annihilation of pathogenic biofilm organisms and in the reduction and elimination of existing biofilms, as well as in the prevention of biofilm formation. The EDTA combinations and compositions of the invention additionally exhibit both anti-protozoal and anti-amimic activity. Based on the published reports, it is further expected that the EDTA compositions of the present invention exhibit anti-viral activity. The EDTA formulations of the present invention are safe for human administration, and are biocompatible and non-corrosive. They may also have anticoagulant properties and are therefore useful for the prevention and / or treatment of a variety of catheter-related infections. The antiseptic solutions of the present invention have numerous applications, including rinsing and washing and rinsing solutions for various types of catheters, use as antiseptic agents or solutions for disinfecting a range of medical, dental and veterinary devices, instruments and other objects, surfaces, and similar. In addition, they have disinfection applications in industrial facilities, and in food handling and preparation.

The antiseptic compositions of the invention can be used prophylactically to prevent infection, or reduce and / or eliminate existing or established infections. Methods for preventing or treating infection of an object or surface with an undesirable microorganism are provided, such methods comprise contacting the surface or object with a composition of the present invention. The compositions of the invention can therefore be employed to inhibit the growth and proliferation of microbial populations and / or fungal pathogens, including the inhibition of biofilm formation and proliferation; inhibit the growth and proliferation of protozoan populations; inhibit the growth and proliferation of amystic populations; and to prevent the infection amíbica, particularly infections by Acanthamoeba. The present invention also provides methods for substantially eradicating microbial populations, including both planktonic microbial populations and microbial populations in the form of biofilms, together with a method for substantially eradicating a population of Acanthamoeba. Such methods comprise contacting an infected object or surface, or an object or surface suspected of being infected, with a composition of the present invention. Depending on the antiseptic composition used in the various methods, various contact time periods may be required to inhibit the formation and proliferation of various populations, and / or substantially eradicate several populations. Appropriate contact time periods for various compositions are provided in the examples and may also be determined by routine experimentation. In one embodiment, the antiseptic compositions of the present invention have at least four, and preferably at least five, of the following properties: anticoagulant properties; inhibitory and / or bactericidal activity against a broad spectrum of bacteria in a planktonic form; inhibitory and / or fungicidal activity against a spectrum of fungal pathogens; inhibitory and / or bactericidal activity against a broad spectrum of bacteria in a sessile form, or biofilm; inhibitory activity against protozoan infections; inhibitory activity against Acanthamoeba infections; safe and biocompatible, at least in modest volumes, in contact with a patient; safe and biocompatible, at least in modest volumes, in a patient's blood stream; and safe and biocompatible with industrial objects and surfaces. Soluble salts of EDTA are used in the compositions of the present invention. Sodium salts of EDTA are commonly available and are generally used, including di-sodium, tri-sodium and tetra-sodium salts, although other salts of EDTA, including ammonium, di-ammonium, potassium, di-potassium, di- Cupric sodium, di-sodium magnesium, ferric sodium, and combinations thereof, may be used, provided they have desired antibacterial, fungicidal, anti-protozoal and / or anti-ammic properties, and that they are sufficiently soluble in the desired solvent. The various combinations of EDTA salts can be used and may be preferred for particular applications. Importantly, in most embodiments, the compositions and methods of the present invention do not employ traditional antibiotic agents and therefore do not contribute to the development of antibiotic-resistant organisms. The antiseptic compositions of the invention comprise, consist of, or consist essentially of, one or more salts of EDTA in solution at a pH higher than the physiological pH. Preferably such compositions have a pH of at least 8.0. Preferably, the compositions of the invention have a pH in the range between 8.5 and 12.5, between 9.5 and 11.5, or between 10.5 and 11.5. The EDTA salt is generally present at an amount of at least 0.01% w / v. In preferred embodiments, the compositions of the invention comprise at least one EDTA salt in an amount of 0.2% to 10% w / v, more preferably 0.2% to 6.0% w / v and most preferably 0.2% to 4.0% w / v. In specific embodiments, the compositions of the invention comprise, or consist essentially of, a solvent and at least one EDTA salt at a concentration of between 0.01% and 10% w / v, wherein the solution has a pH of at least 9.0 and possesses bactericidal activity against a broad spectrum of bacteria. Preferably, at least one salt of EDTA is tri-sodium or tetra-sodium EDTA. The solvent is generally selected from the group consisting of: water, saline, alcohols, and combinations thereof. In one embodiment, the solvent is a combination of water and ethanol. The antiseptic compositions of the invention can be used as rinsing and rinsing solutions for various types of resident access catheters, including vascular catheters used for the delivery of fluids, blood products, drugs and nutrition, fluid or blood extraction, dialysis , monitoring of patient conditions, and the like. The antiseptic solutions of the present invention can also be used as rinsing and washing and rinsing solutions for urinary catheters, nasal tubes, mouth tubes, and the like. The general solution parameters described below are suitable for these purposes. In one embodiment, an antiseptic solution comprising, consisting of or consisting essentially of one or more salts of sodium EDTA at a pH higher than the physiological pH is employed to maintain the opening of resident intravascular access devices. Methods of disinfecting catheters and other medical tubes, such as nasal tubes, mouth tubes and the like, are also provided and involve contacting the catheter or other medical tube with a disinfectant composition of the present invention. In another embodiment, the antiseptic compositions of the present invention which comprise, consist of or consist essentially of one or more sodium salts of EDTA at a pH higher than the physiological pH are provided as disinfecting solutions for medical devices such as dentures and other devices dental, orthodontic and / or periodontal, for contact lenses and other optical devices, for instruments, medical and veterinary devices, and the like, and as disinfectant solutions for disinfecting surfaces and objects. Methods of disinfecting such devices are also provided, the methods comprising contacting a device with an antiseptic composition of the present invention. In general, the antiseptic compositions of the present invention can be used as soaking solutions for dental, orthodontic and periodontal devices, including toothbrushes, and can also be used as soaking solutions for contact lenses and other optical devices, as well as instruments, medical and veterinary devices and the like. For these applications, the antiseptic compositions of the present invention are generally formulated as solutions, or are provided in a dry form which, in the addition of a suitable solvent, forms a solution. In yet another embodiment, the antiseptic compositions of the present invention are formulated for use in solutions, gels, creams and other preparations designated for topical use as antiseptic agents, wipes, antibacterial treatments and the like. The antiseptic compositions of the present invention can also be used as antibacterial agents in connection with bandages, dressings, wound healing agents and devices, and the like. In related embodiments, covers for use in wound healing, such as bandages and dressings, are provided wherein the cover is impregnated with one or more of the compositions of the present invention. The antiseptic compositions of the present invention can also be used in industrial facilities such as water storage and distribution systems, water purification, humidification and dehumidification devices, and in preparation, handling and food packaging facilities, to inhibit, reduce or substantially eliminate microbial populations in both free and sessile floating forms, as well as many fungal, amoebic, and protozoal populations. Industrial surfaces and equipment may be contacted with, jetted with, or soaked in, antiseptic compositions of the present invention. Formulations of antiseptic release composition with time may also be provided to provide the overtime treatment, particularly in locations that are difficult to access frequently.

BRIEF DESCRIPTION OF THE FIGURES Figures 1A-1D show the minimum inhibitory concentration (CM) and minimum bactericidal concentrations (MBC) for several gram-positive and gram-negative bacterial organisms against EDTA salt solutions consisting essentially of: EDTA di- potassium, - di-ammonium EDTA; EDTA di-sodium; EDTA tri-sodium; and tetra-sodium EDTA, using the agar dilution method. Bacterial organisms were isolated from catheter-related infections in human patients. The experimental techniques are described in Example 1. Figure 2 shows the MIC and MBC concentrations for several fungal organisms against different formulations of EDTA, using the agar dilution method.

The experimental techniques are described in Example 1. Fungal organisms were collected from human patient samples. Figures 3A and 3B show CIM and CBM data for gram-positive and gram-negative bacterial organisms against EDTA salt solutions consisting essentially of: cupric di-sodium EDTA; EDTA di-sodium magnesium; and ferric sodium EDTA. Bacterial organisms were isolated from catheter-related infections in human patients. The experimental techniques are described in Example 1. Figures 4A-4C show CIM and CBM data for several gram-positive and gram-negative bacterial organisms against EDTA salt solutions in combination consisting essentially of: cupric di-sodium EDTA and tetra-sodium; Di-sodium sodium cupric and di-potassium; and di-sodium EDTA, cupric and di-ammonium. Bacterial organisms were isolated from catheter-related infections in human patients. The experimental techniques are described in Example 1. Figures 5A-5C show CIM and CBM data for several gram-positive and gram-negative bacterial organisms against EDTA salt solutions in combination consisting essentially of: tetra-sodium EDTA and di-ammonium; Tetra-sodium and di-potassium EDTA; and di-ammonium and di-potassium EDTA. Bacterial organisms were isolated from catheter-related infections in human patients. The experimental techniques are described in Example 1. Figure 6 shows the minimum biofilm eradication concentration (CEBM) values for various organisms, expressed in mg / ml of tetra-sodium EDTA (w / v) using the methodology described in Example 2. Figure 7 shows the experimental results of the treatment of renal hemodialysis catheters infected with an antiseptic composition consisting essentially of tetra-sodium EDTA at a concentration of 40 mg / ml (w / v). Figure 8 shows the experimental results of the treatment of infected renal hemodialysis catheters, as well as an arterial and a venous catheter, with an antiseptic composition consisting essentially of tetra-sodium EDTA at a concentration of 20-100 mg / ml (p / v).

DETAILED DESCRIPTION OF THE INVENTION EDTA is used at low concentrations in many compositions, in combination with other active components, as a stabilizing or preservative agent. The antiseptic compositions of the present invention generally comprise higher concentrations of EDTA and preferably comprise at least 0.01% of EDTA salts by weight per volume of solution (w / v), and may comprise up to 15% (w / v) of salts of EDTA. For many applications, the antiseptic compositions of the invention preferably comprise at least 0.1% (w / v) EDTA salts and less than 10% (w / v) EDTA salts, more preferably between 0.1 (w / v) salts of EDTA and 8.0% (w / v) of EDTA salts, and most preferably between 0.1% and 6.0% of EDTA salts. Exemplary compositions, described below, comprise 3.6-4.4% (w / v) of EDTA salts in aqueous solution, or 0.01-0.2% (w / v) of EDTA salts in a mixture of water and ethanol. The concentration of EDTA salts described for various applications may depend on the salt of EDTA, or combination of salts, employed, the type of infection to be treated and, to some degree, the solvent used for the antiseptic compositions. For example, when aqueous solvents comprising ethanol are used, the concentration of EDTA salts required to provide the desired level of activity can be reduced compared to the concentration of EDTA salts used in compositions having water as the solvent. Antiseptic compositions comprising one or more EDTA salts have demonstrated inhibitory and / or bactericidal efficacy at concentration ranges of 0.01% to 30% or more, as shown in the exemplary data provided below. The "effective" concentrations of EDTA salts in the antiseptic compositions of the present invention for inhibitory, bactericidal, fungicidal, biofilm eradication and other purposes can be determined by routine experimentation, as described in the examples provided below. The British Pharmacopoeia (FB) specifies that a 5% solution of di-sodium EDTA has a pH of 4.0 to 5.5. The FB also specifies a pH range of 7.0 to 8.0 for tri-sodium EDTA solutions. The pH values for other EDTA salts in aqueous solution are given in Example 10 below. At physiological pH, sodium salts of EDTA exist as a combination of di-sodium and trisodium EDTA, with the tri-sodium salt of EDTA being predominant. In the United States, "di-sodium" pharmaceutical EDTA prepared for injection has generally been titrated with sodium hydroxide at a pH of 6.5 to 7.5. At this pH, the EDTA solution currently comprises mainly tri-sodium EDTA, with a minor proportion of the di-sodium salt. Other compositions comprising sodium salts of EDTA that are used in medical or health care applications generally conform to a pH that is substantially physiological. In certain embodiments, the antiseptic compositions of the present invention comprise, consist essentially of, or consist of, a salt of sodium EDTA (or a combination of salts of sodium EDTA) in solution at a pH greater than physiological, preferably at a higher pH 8.0, at a pH greater than 8.5, at a pH greater than 9, at a pH greater than 9.5, or at a pH greater than 10. In other embodiments, the antiseptic compositions of the present invention comprise, consist essentially of, or consist of of, a salt of sodium EDTA (or a combination of sodium EDTA salts) in solution at a pH in the range between 8.5 and 12.5, at a pH between 9.5 and 11.5, or at a pH between 10.5 and 11.5. When used herein, the term "EDTA salt" may refer to a single salt, such as di-sodium, tri-sodium or tetra-sodium salt, or another form of EDTA salt, or may refer to a combination of such salts. The composition of EDTA salts depends both on the EDTA salts used to formulate the composition, and on the pH of the composition. For antiseptic compositions of the present invention comprising sodium EDTA salts at the desired pH ranges (specified above), sodium EDTA salts are predominantly present in both tri-sodium and tetra-sodium salt forms. In one embodiment, the antiseptic compositions of the present invention comprise, or consist essentially of, a combination of at least the tri-sodium and tetra-sodium salts of EDTA. In another embodiment, the antiseptic compositions of the present invention comprise, or consist essentially of, a combination of at least the tri-sodium and tetra-sodium salts of EDTA, in which at least 10% of the EDTA in the composition is present. in the tetra-sodium salt form. In yet other embodiments, the antiseptic compositions of the present invention comprise, or consist essentially of, a combination of at least tri-sodium and tetra-sodium salts of EDTA, in which at least 50% or at least 60% of EDTA in the composition it is present in the tri-sodium salt form. In another modality, the antiseptic compositions of the present invention comprise, or consist essentially of, a combination of di-sodium, tri-sodium and tetra-sodium EDTA, in which less than 10% of the EDTA in the composition is present in the salt form of di-sodium. The antiseptic compositions comprising, consist essentially of, or consist of EDTA salts other than, or in addition to, sodium EDTA salts have different "effective" pH ranges. The "effective" pH ranges for EDTA salts desired in antiseptic compositions of the present invention for use in inhibitory, bactericidal, fungicidal, biofilm eradication and other purposes, can be determined by routine experimentation. In some embodiments, the antiseptic compositions of the present invention consist of the EDTA salts, as described above, and the antiseptic solutions consist of EDTA salts dissolved in a solvent, generally an aqueous solvent such as water or saline. In other embodiments, the antiseptic compositions of the present invention consist essentially of the EDTA salts, as described above, generally in an aqueous solvent such as water or saline. The antiseptic compositions of the present invention consisting essentially of an EDTA salt, or a combination of EDTA salts, are substantially free of other active substances having substantial antimicrobial and / or anti-fungal activity. Substantial antimicrobial and / or anti-fungal activity, in this context, means the antimicrobial and / or anti-fungal activity ie at least 50% of the antimicrobial and / or anti-fungal activity of a sodium EDTA salt composition in aqueous solution at a concentration of 4.0% (w / v) at a pH of 10.5. In some embodiments, the antiseptic compositions of the present invention comprise EDTA salts having specific concentrations, at specific pH ranges, and may contain materials, including active components, in addition to the EDTA salts described above. Other antimicrobial or biocidal components may be incorporated into the antiseptic compositions of the present invention, although the use of traditional antibiotics and biocidal agents is generally disapproved as a consequence of the frightening consequences of the development of organisms resistant to antibiotics and biocides. In some embodiments, the antiseptic compositions of the present invention comprising EDTA salts at specific concentrations and pH ranges, are substantially free of other active substances having substantial antimicrobial and / or anti-fungal activity. Other active or inactive components can also be incorporated into the antiseptic compositions of the present invention, provided they do not deleteriously affect the activity and / or stability of the EDTA salts. The proteolytic agents can be incorporated into the antiseptic compositions for some applications. Antiseptic compositions formulated for topical application may include various creams, emoluments, and skin care compositions such as aloe vera, and the like, for example. The antiseptic compositions of the present invention provided in a solution formulation may also comprise other active and inactive components, so long as they do not interfere negatively with the activity and / or stability of the EDTA salts. The compositions of the present invention can be used in a dry form or solution. In solution, the EDTA salts are preferably dissolved in a solvent, which may comprise an aqueous solution, such as water or saline, or another biocompatible solution in which the EDTA salts are soluble. Other solvents, including alcohol solutions, can also be used. In one embodiment, the EDTA salt compositions of the present invention are formulated in a mixture of water and ethanol. Such solutions are highly effective and can be prepared by producing a stock solution of EDTA salts concentrated in water and then introducing the desired concentration of ethanol. The EDTA salt concentrations of approximately 0.01% to 10%, p / v, are suitable, and ethanol concentrations of between about 0.1% and about 10%, v / v, provide effective antiseptic compositions. In some embodiments, EDTA salt concentrations of about 2 mg / ml (0.2% w / v) in water with an ethanol concentration of about 1% (v / v) are highly effective against a broad spectrum of bacterial strains. When the sodium EDTA salts are used, the pH ranges of these antiseptic compositions are as described above. Non-aqueous biocompatible solvents can also be used, provided that the EDTA salts can be solubilized and remain in solution during storage and use. The EDTA solutions of the present invention are preferably provided in a sterile, non-pyrogenic form, and can be packaged in any convenient manner. In some embodiments, the EDTA antiseptic compositions of the present invention may be provided in connection with or as part of a medical device, such as in a pre-filled syringe or other medical device. The compositions can be prepared under sterile, aseptic conditions, or can be sterilized after preparation and / or packaging using any of a variety of suitable sterilization techniques. The single or multiple use of vials, syringes or containers of EDTA solutions can be provided. The systems of the present invention include such vials, syringes or containers containing EDTA solutions of the present invention. The compositions of the present invention can also be provided in a substantially "dry" form, such as a substantially dry coating on a pipe surface or a conduit, or a medical or industrial device such as a catheter or conduit, or a container, or similar. Such substantially dry forms of the EDTA compositions of the invention can be provided in a powder or lyophilized form that can be reconstituted to form a solution with the addition of a solvent. The substantially dry forms of the EDTA compositions may alternatively be provided as a coating, or they may be incorporated in a gel or other type of carrier, or encapsulated, or otherwise packaged, and provided on a surface as a coating or in a a container. Such substantially dry forms of the EDTA compositions of the invention are formulated so that, in the presence of a solution, the substantially dry composition forms an EDTA solution having the composition and properties described above. In certain embodiments, different encapsulation or storage techniques can be employed so that the EDTA effective time release is carried out on prolonged exposure to the solutions. In this embodiment, substantially dry EDTA solutions can provide antiseptic activity for a prolonged period of time and / or multiple exposures to the solutions. Compositions comprising EDTA have a well-established safety profile in connection with medical use and administration to humans. Doses up to 3000 mg of disodium EDTA are drained for 3 hours, on a daily basis, for the treatment of hypercalcemia in humans and are well tolerated. EDTA salts are also present, in combination with other components, in many solutions used in human and medical health applications, and have been established as safe for human use, both in vitro and in vivo, EDTA salts are easily available at a reasonable cost, and extra time in solution is stable. The formulation and production of antiseptic compositions of the present invention are generally direct. In one embodiment, the desired antiseptic compositions of the present invention are formulated by dissolving at least one EDTA salt in an aqueous solvent, such as purified water., at the desired concentration and adjusting the pH of the EDTA salt solution to the desired pH. In alternative embodiments, the desired antiseptic compositions of the present invention are formulated by dissolving at least one EDTA salt in a solvent in which the EDTA salt, or combination of salts, is soluble to provide a concentrated, solubilized EDTA salt solution. . Additional components or solvents can then be added. Alternatively, the solubilized EDTA salt composition can be formulated in a different form from a solution, such as a topical preparation. The antiseptic solution can then be sterilized using conventional means, such as autoclave, UV irradiation, filtration, ultrafiltration, and / or other means. The preferred osmolarity range for EDTA solutions is 240-500 OsM / kg, more preferably 300-420 mOsm / kg. The solutions are preferably formulated using USP materials. Antiseptic compositions comprising, consisting of, or consisting essentially of, tri- or tetra-sodium salts, or a mixture thereof, are preferred for many applications and can be prepared using EDTA sodium salts other than tri-sodium salts. - and tetra-sodium, such as di-sodium EDTA which is readily available. The di-sodium EDT solutions have a lower pH in solution than the desired pH range of the compositions of the present invention. However, in adjusting the pH to the desired range using a pH adjusting material, such as sodium hydroxide, sodium acetate or other well-known pH adjusting agents, the EDTA solutions prepared using di-sodium salts are convert to the EDTA compositions of preferred di-, tri- and / or tetra-sodium salt of the present invention. Accordingly, different forms and combinations of EDTA salts can be used in the preparation of the EDTA compositions of the present invention, provided that the pH of the composition is adjusted to the desired pH range prior to use. In one embodiment, antiseptic compositions consisting of a mixture primarily of tri- and tetra-sodium EDTA are provided by dissolving di-sodium EDTA in an aqueous solution in an amount of 3% -5% on a weight / volume basis, and adding hydroxide of sodium in an amount sufficient to provide the desired pH of between 8.5 and 12.0. The antiseptic compositions of the present invention which comprise, consist essentially of, or consist of, at least one EDTA salt as described above are also useful for many other applications. EDTA solutions can be used as antiseptic solutions for soaking, rinsing, or contacting medical, dental and veterinary surfaces and objects. The EDTA solutions of the present invention can be used, for example, to store and / or disinfect contact lenses and other optical devices; for storing and / or disinfecting dental devices such as dentures, bridges, retainers, toothbrushes, and the like; and to store and / or disinfect medical, dental and veterinary devices and instruments. In these applications, the devices or surfaces may be contacted with, or soaked in, EDTA solutions of the present invention for a sufficient time to substantially eliminate microbial and / or fungal infections. The EDTA compositions of the present invention can additionally be used to disinfect water and other fluid supply lines. The disinfection of fluid supply lines can be performed by intermittently washing the lines with EDTA compositions of the present invention. Similarly, the EDTA compositions of the present invention can be used to eradicate biofilms, and microbial (including some viruses and protozoa) and fungal populations in water storage and delivery devices. Numerous methods and experimental tests have been performed using EDTA-containing compositions of the present invention to establish their properties and their effectiveness as antiseptic compositions. Various experimental procedures are described in detail later. These procedures and experimental results are provided for illustrative purposes only and are not intended to limit the scope of the present invention in any way.

Example 1 Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) data for organisms against different formulations of EDTA, using the agar dilution method Minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) for various organisms yeast and gram-positive and gram-negative bacteria were established for various different formulations of EDTA using the agar dilution method described below. The CIM and MBC for several organisms were also tested in combinations of salts of EDTA The gram-positive and gram-negative bacterial organisms were isolated from human patients having catheter-related infections to ensure that the bacterial strains were actively pathogenic and were of the type common in bacterial infections related to human catheters. Yeast organisms were collected from patients who have serious septicemic infections. The organisms were cataloged and maintained in Peter Kite's laboratory at the University of Leeds. Several EDTA salt solutions and EDTA salt solutions in combination were prepared by dissolving relevant reagent grade EDTA salts in distilled water at the desired concentration (w / v) of EDTA salt. Concentrated mother EDTA salt solutions were prepared for each EDTA salt or EDTA salt combination and used to determine CIM and CBM for various organisms. The tetra- and tri-sodium EDT solutions were prepared using the tetra- and tri-sodium salts of EDTA rather than using di-sodium EDTA and adjusting the pH of the solution to achieve the desired pH ranges. The EDTA salt solutions were sterilized prior to use and stored at 4 ° C.

Dilution method protocol on agar Production of agar • Place 2 liters of nutrient agar in a steam bath and leave for about 1 hour (until melting). • Allow the agar to cool to 50 ° C. • Collect 20 sterile glass bottles (125 ml) and distribute 100 ml of the nutrient agar to each one. To these add 0.5, 1.0, 1.5, 2.0, 4, 6, 8, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90 and 100 mg / ml of tetra-sodium EDTA (u another salt of EDTA or combination of EDTA salt to be tested), using a stock solution at 200 mg / ml. • Pour 200 ml of agar into a sterile petri dish and allow settling. Pour 3 additional plates.

Label the plates with the concentration of EDTA they contain. Do this for each concentration. • These plates are then stored, until needed, in a refrigerator at 4 ° C.

Inoculation of the plates • Grow the cultures of 23 gram-positive organisms and 19 gram-negative organisms in nutrient broth overnight. • Dilute each culture to 106 cfu / ml, using phosphate buffered saline (PBS). • Use an automatic plate inoculator to inoculate each plate with 21 organisms. • Incubate plates overnight at 37 ° C. • The next day register + or - for growth.

• Use sterile filter paper to transfer the growth of the initial plates to new Cled agar plates to determine the CBM. • Incubate the duplicate plates overnight at 37 ° C. • The next day register + or - for growth. The CIM and MBC were described as the lowest concentration at which there was no growth. The results are shown in Figures 1A-5C. Figures 1A-1D show CIM and CBM data (reported as mg / ml EDTA solution, w / v) for many gram-positive and gram-negative organisms against EDTA salt solutions consisting essentially of: EDTA di- potassium Di-ammonium EDTA; EDTA di-sodium; Tri-sodium EDTA and tetra-sodium EDTA. Figure 2 shows data of MIC and MBC (presented as mg / ml EDTA solution, w / v) for yeast against EDTA salt solutions consisting essentially of: tetra-sodium EDTA; DI-potassium EDTA; and di-ammonium EDTA. Figures 3A and 3B show CIM and CBM data (presented as mg / ml EDTA solution, w / v) for gram-positive and gram-negative organisms against EDTA salt solutions consisting essentially of: di-sodium EDTA cupric; EDTA di-sodium magnesium and di-sodium EDTA; Tri-sodium EDTA and ferric sodium EDTA. Figures 4A-4C show CIM and CBM data (presented as mg / ml EDTA solution, w / v) for gram-positive and gram-negative organisms against EDTA salt solutions in combination consisting essentially of: EDTA di -sodic copper and tetra-sodium; Di-sodium sodium cupric and di-potassium; and di-sodium EDTA, cupric and di-ammonium. Figures 5A-5C show CIM and CBM data (presented as mg / ml EDTA solution, w / v) for gram-positive and gram-negative organisms against EDTA salt solutions in combination consisting essentially of: EDTA tetra -sodium and di-ammonium; Tetra-sodium and di-potassium EDTA; and di-ammonium and di-potassium EDTA. Various EDTA salts and EDTA salt combinations were effective in inhibiting and / or eliminating a broad spectrum of bacterial strains at reasonable concentrations. The use and previous medical test have established good biocompatibility profiles for the use of sodium EDTA salts in humans and animals, while the biocompatibility of other EDTA salts has not yet been established. The salts of tetra- and tri-sodium EDTA appear to be more effective against a broad spectrum of pathogenic bacteria. In addition, they have, or may easily be, established to be biocompatible for human and veterinary use, and are cost-effective and stable. The tetra-sodium EDTA salt is additionally active as an anticoagulant and is highly soluble in aqueous solvents. Based on these factors and the experiments summarized above, the salts of tetra- and tri-sodium EDTA were chosen as the most promising candidates for antiseptic compositions of the present invention.

Example 2 Minimum biofilm eradication concentration (CEBM) data for organisms against Tetra-sodium EDTA, using the modified Calgary device method Biofilm formation is an important factor in bacterial contamination. An effective antiseptic composition preferably has the ability to reduce biofilm proliferation, or prevent or inhibit the formation of biofilms. Therefore, our antiseptic solution of tetra-sodium EDTA candidate was tested to determine whether it can prevent or inhibit the formation of biofilms. The minimum biofilm eradication concentration (CEBM) for several organisms against tetra-sodium EDTA was established using a modified Calgary device method. The Calgary method is described in Olsen et al. , Canadian Journal of Veterinary Research, 66: 86-92 (2002) and in United States Patent 6,599,714. The method and results are described later. Salt solutions of tetra-sodium EDTA were prepared by dissolving the salt of EDTA tetra-sodium reagent grade in distilled water at the concentration (w / v) of the desired EDTA salt. Concentrated tetra-sodium EDTA salt solutions were prepared to determine the CEBM for various organisms in a sessile form, or biofilm. The tetra-sodium EDTA solutions were sterilized prior to use and stored at 4 ° C.

Method Biofilm formation: • Use 100 ml Muller Hinton broth of the required organism during the night. • Pipette 200 μl into all wells in a 96-well microtiter tray. Place a lid with 96 pins. Incubate in an orbital shaker for 24 hours at 37 ° C at a speed of 200 rpm.

Susceptibility test: • Use previously formed biofilm. • Place cap (with pins) in a new 96-well microtiter tray containing 250 μl of required concentrations of test agent. Incubate for 1 to 24 hours at 37 ° C. (Not in shaker). • At time intervals of 1, 3, 6 and 24 hours, remove 4 pins for each concentration of the cap by inserting a screwdriver and quickly disconnecting the pin in the cavity. • Place 3 pins for each concentration in a 5 ml wash of PBS and invert once. • Place the three pins in 3 ml of PBS and sonicate for 15 minutes. Deposit 2 μl on 3X CLED plates and spread using a sterile plastic spreader. Incubate at 37 ° C overnight. Read the colony counts the next day. • Place the remaining loose pin (for each concentration) in 600 μl of 4% formal saline solution for SEM. The CEBM values for several organisms, expressed in mg / ml of tetra-sodium EDTA (w / v) determined using this method, are shown in figure 6. The results show that 40 mg / ml of tetra-sodium EDTA (4 % p / v) was an effective biofilm eradication concentration for all tested microbial populations. The exemplary data generated by the CEBM experiments for various microorganisms are provided below. Tetra-sodium EDTA was used for all experiments, which were performed in triplicate.

Table 1: Organism: 250 E. col ± For 250 E. coli, CEBM = 10 mg / ml tetra-sodium EDTA. Table 2: Organism: J26 Pseudomonas aeruginosa For J26 Pseudomonas aeruginosa, the CEBM = < 5 mg / ml tetra-sodium EDTA.

Table 3: Organism: 292 JEJnterobacter cloacae For 292 Enterobacter cloacae, the CEBM = < 5 mg / ml tetra-sodium EDTA. Table 4: Organization: ff. Enterococcus sp.

For H. Enterococcus sp., The CEBM = < 5 mg / ml tetra-sodium EDTA.

Table 5: Organism: J22 Enterobacter cloacae For J22 Enterobacter cloacae, CEBM = 15 mg / ml tetra-sodium EDTA. Table 6: Organism: R81 Proteus vulgar ± s For R81 Proteus vulgaris, the CEBM = < 5 mg / ml tetra-sodium EDTA.

EXAMPLE 3 In Vitro Catheter Flushing Treatment Procedure in Patient Positive Catheters A catheter flushing treatment procedure using 40 mg / ml (4% w / V) of tetra-sodium EDTA candidate solution was developed and used for catheters of sample patient hemodialysis that proved to be positive for several microbial infections. Catheters that were determined to have microbial infections were subjected to catheter rinsing treatment using tetra-sodium EDTA, and the colony counts were taken at several time points. In a first experiment, all catheters were treated with 4% w / v tetra-sodium EDTA solution while in a second set, the catheters were treated with tetra-sodium EDTA solutions at various concentrations. Tetra-sodium EDTA solutions were prepared and stored as described above in Examples 1 and 2. The procedure and results are described below.

Method: • Renal hemodialysis catheters removed on suspicion of infection were selected, flushing with 1 ml of saline buffered with sterile phosphate under each lumen. The quantitative culture was performed using aliquots of 1 and 10 uL spread on blood agar plates and incubated. • The catheters were initially stored at 4 ° C until after the selection and the external lumen was sterilized with an alcohol wipe. • Prior to the rinse treatment test, selected positive catheters were rinsed with nutrient broth using a 5 ml syringe and incubated overnight at 37 ° C to ensure biofilm viability and to ensure complete colonization of all endoluminal surfaces with the infection organism. • After the overnight incubation, each catheter lumen was flushed with 5 ml of sterile saline and two 1 cm pieces were cut from the distal end, each placed in 1 ml of 1M sterile calcium chloride, (for neutralization of the agent) one for scanning electron microscopy (MEE) and the other for culture, in sterile universal containers. • For the culture procedure, the universal was placed in a sonication bath for 15 min at room temperature and then vortexed for 20 seconds.

• Quantitative culture was performed using 1 μl and 10 μl aliquots plated on blood agar plates and strewn with sterile plastic L-shaped rods, incubated at 37 ° C overnight, and colonies They told the next day. • The catheter was flushed and rinsed with the appropriate concentration of tetra-sodium EDTA rinse fluid and incubated at 37 ° C for 18 hours. • At 3, 6 and 18 hours of incubation, two 1 cm pieces of the end of the catheter were cut and neutralized in 1 ml of 1M sterile calcium chloride solution. • The quantitative counting procedure was followed, at each time interval, as previously described and one piece was retained for SEM. Seventeen (17) infected renal hemodialysis catheters were treated with an antiseptic composition consisting of tetra-sodium EDTA at a concentration of 40 mg / ml (4% w / v). The results are shown in Figure 7. Ten additional infected renal hemodialysis catheters as well as an arterial and a venous catheter were treated with an antiseptic composition consisting of tetra-sodium EDTA at concentrations of 20-100 mg / ml (2- 10% p / v). The results are shown in figure 8. The results show that 40 mg / ml (4% w / v) of tetra-sodium EDTA are effective in the annihilation, or dramatic reduction, of the population of most organisms after a 24 hour treatment. This concentration of tetra-sodium EDTA is safe for use in connection with humans and other animals and is considered to be effective, and therefore is a desired concentration for antiseptic compositions and methods of the present invention.

Example 4 The effect of tetra-sodium EDTA on Acanthamoeba and the effect of Klebsiella treated with tetra-sodium EDTA on Acanthamoeba. Various Acanthamoeba species are capable of infecting humans. Acantamic infections frequently result as a consequence of the inappropriate storage of contact lenses and other medical devices that come into contact with the human body. Acantamebas feed on bacterial populations and are resistant to many treatments. The effect of tetra-sodium EDTA, prepared as described above, in Acanthamoeba populations was tested as follows. Tetra-sodium EDTA compositions were also prepared using Page saline and physiological saline as solvents. The effect of Klebsiella treated with tetra-sodium EDTA on Acanthamoeba was also tested experimentally using the following methodology.

The effect of tetra-sodium EDTA on Acanthamoeba Method: • Incubate a fresh blood agar plate with Klebsiella edwardsii at 37 ° C 18 hours prior to the test. • Use a stock solution of EDTA tetra-sodium (100 mg / ml), make a concentration of 22 and 44 mg / ml in Page saline. • Place 9 ml of each concentration in a sterile glass test tube. Place 9 ml of sterile Page saline solution in another sterile glass test tube to act as a control. • Make a suspension of Klebsiella edwardsii in 6 ml of Page sterile saline. Adjust to standard 5 of McFarland. • Add 1 ml of suspension to each serial dilution and control. Due to the dilution factor of the Klebsiella suspension each concentration will now be at 20 and 40 mg / ml. The control still contains non-tetra sodium EDTA. Repeat all concentrations in physiological saline. • Submit to vortex to mix. Each tube should now contain a suspension of Klebsiella at 0.5 McFarland. • Scrape the entire surface of the Acanthamoeba plate and suspend in 1.5 ml of Page's saline. Submit to vortex. • Add 200 ul of the Acanthamoeba suspension to each serial dilution and control. • Place the test tubes in an incubator at 30 ° C for 24 hours. After the incubation centrifuge each universal by minutes at 3000 rpm. Pour the supernatant and resuspend the pellet. Place 10 uL in duplicate of each dilution and control on a non-nutrient agar plate with a Klebsiella sieve. Cut a slot below the center of each plate to prevent migration and place 10 uL of the dilution tested on each side.

Mark each inoculation site with a black marker pen.

Incubate the plates for 72 hours at 30 ° C. Verify the growth of Acanthamoeba by direct visualization of the plates using an X10 magnifying eye microscope, start at each inoculation site.

Table 7: Growth after 24 hours of incubation with tetra-sodium EDTA Table 8: Growth after 24 hours of incubation with tetra-sodium EDTA (repetition) Table 9: Growth after 48 hours of incubation with tetra-sodium EDTA The results show that 20-40 mg / ml (2-4% w / v) of tetra-sodium EDTA in Page saline and physiological is effective to reduce, or substantially eliminate, populations of Acanthamoeba after 48 hours of exposure. Tetra-sodium EDTA compositions prepared using water as the solvent were also effective (data not shown). These results indicate that the antiseptic compositions of the present invention are suitable for application as soaking solutions for various medical devices and instruments, including contact lenses, and dental, orthodontic and / or periodontal devices. The antiseptic compositions of the present invention are also effective in substantially eliminating Acanthamoeba populations in other applications, including in freshwater and seawater distribution and storage systems, in heating, venting and air conditioning units, humidifiers, dialysis units. , and similar. The Acantiíamsejba feed on bacterial populations. It was therefore tested whether a bacterial population that was treated with antiseptic EDTA compositions of the present invention could have some effect on Acanthamoeba that are fed into the treated bacterial population.

The Klebsiella effect is treated with tetra-sodium EDTA in Acanthamoeba Method: • Incubate a fresh blood agar plate with Klebsiella edwardsii at 37 ° C 18 hours prior to the test. • Use a stock solution of EDTA tetra-sodium (100 mg / ml), make a concentration of 22 and 44 mg / ml in Page saline solution. • Place 9 ml of each concentration in a sterile glass test tube. Place 9 ml of sterile Page saline solution in another sterile glass test tube to act as a control. • Make a suspension of Klebsiella edwardsii in 6 ml of Page sterile saline. Adjust to standard 5 of McFarland. • Add 1 ml of suspension to each serial dilution and control. Due to the dilution factor of the Klebsiella suspension each concentration will now be at 20 and 40 mg / ml. The control still contains non-tetra sodium EDTA. Repeat all concentrations in physiological saline. • Submit to vortex to mix. Each tube should now contain a suspension of Klebsiella at 0.5 McFarland.

• Incubate the tubes at 37 ° C overnight. • The next day, centrifuge the tubes at 300 rpm for 10 minutes. Pour the supernatant; add 10 ml of saline or new Page saline solution, resuspend and re-centrifuge. Pour the supernatant and resuspend in 1 ml of either saline or Page saline solution. • Scrape the entire surface of the Acanthamoeba plate and suspend in 1.5 ml of Page's saline. Submit to vortex. • Add 200 ul of the Acanthamoeba suspension to 3 tubes containing 9 ml of saline and 3 tubes containing 3 ml of Page's solution. Label each tube as if they were the concentrations of EDTA used in the incubation with Klebsiella. • Add 1 ml of resuspended Klebsiella to the appropriate tube containing Acanthamoeba. • Place the test tubes in an incubator at 30 ° C for 24 hours. • Accommodate other sets of tubes to incubate Klebsiella with EDTA at 37 ° C, overnight as before. • After incubation, centrifuge each tube containing Acanthamoeba for 10 minutes at 3000 rpm. • Pour the supernatant and resuspend the pellet. • Place 10 uL in duplicate of each dilution and control on a non-nutrient agar plate with a Klebsiella sieve (not incubated with EDTA). Cut a slot below the center of each plate to prevent migration and place 10 uL of the dilution tested on each side. • Mark each inoculation site with a black marker pen. • Incubate plates for 72 hours at 30 ° C.

• Verify the growth of Acanthamoeba by direct visualization of the plates using an X10 magnifying light microscope, start at each inoculation site. • Place the remaining Acanthamoeba suspension in a new set of tubes containing either new saline solution or new saline solution • Wash and resuspend Klebsiella, which has been incubated overnight with EDTA, as before and add to each appropriate tube containing the Acanthamoeba • Incubate the tubes at 30 ° C overnight • After incubation centrifuge each universal for 10 minutes at 3000 rpm • Pour the supernatant and resuspend the pellet • Place 10 uL in duplicate of each dilution and control on a non-nutrient agar plate with a Klebsiella sieve (not incubated with EDTA) Cut a groove below the center of each plate to prevent migration and place 10 μL of the dilution that is test on each side • Mark each inoculation site with a black marker pen.

• Incubate plates at 30 ° C. • Verify the growth of Acanthamoeba by direct visualization of the plates using an X10 magnifying light microscope, start at each inoculation site.

Table 10: Growth of Acanthamoeba after 24 hours of incubation with Klebsiella (previously incubated with EDTA) Table 11: Growth of Acanthamoeba after 48 hours of incubation with Klebsiella (previously incubated with EDTA) These results demonstrate that the growth of Acanthamoeba can be stopped and Acanthamoeba populations can be substantially eliminated from the bacterial populations in which they are fed with the antiseptic EDTA compositions of the present invention. Antiseptic EDTA compositions having a tetra-sodium EDTA concentration of 20-40 mg / ml (2-4% w / v) were effective. This proves the utility of the antiseptic compositions of the present invention for applications such as soaking solutions for various medical devices and instruments, including contact lenses and dental / orthodontic / periodontic devices, as well as for other applications such as storage and distribution systems of fresh water and seawater, in heating, venting and air conditioning units, humidifiers, dialysis units, and the like.

Example 5 Experiments were conducted to determine whether the tetra-sodium EDTA compositions prevent the binding of, and adhesion to, silicon tubing of microorganisms. If the binding and adhesion to silicon tubing of microorganisms can be prevented, the formation of biofilms can be reduced. The experimental protocol used and the results obtained are provided later.

Method: • Fill 1 cm sections of silicon tubing with molten wax to seal each endolumen, harden at 4 ° C.

• Place 4 sections in 30 ml of Phosphate buffered saline (PBS) as a control. Place 8 sections in 30 ml of 4% tetra-sodium EDTA. • After 30 minutes, place the 4 sections of the PBS and 4 sections of the 4% tetra-sodium EDTA in clean containers in a hot block, and allow to dry. • Transfer the remaining 4 sections in 30 ml of sterile PBS to rinse, then allow to air dry as before. • Once dry, place all 12 sections in mixed organisms 105cfu / ml (overnight cultures of Klebsiella penumoniae and CNS allow to grow in nutrient broth at 37 ° C), incubate at 37 ° C. • After 30 minutes, remove 2 sections of each type and rinse in 2 X 30 ml of sterile PBS. Dry with air as before. Use separate washing and drying containers to prevent each type of contamination. • Place each section in 1 ml of PBS in a centrifuge tube, undergo sonication in a sonication water bath for 15 minutes. • Deposit each tube, in duplicate, in the automatic plate inoculator, 50 ul in a log dilution. • Deposit duplicates of each diluted tube 1/10.

• Incubate plates at 37 ° C overnight. Read the colony count in a ProtoCOL automatic plate reader. Repeat after 6 hours. The results for the control catheter sections and EDTA treated are shown below.

Table 12 The results for the pure EDTA solution were found to be more reproducible, and these, therefore, were further analyzed. When the sections were placed in 1 ml of solution, the counts per ml were equal to counts per section.

Table 13 Table 14 Repeat for 24 hours with Klebsiella + CNS: Table 15 Table 16 + Indicates increase in average cfu / control section Results for Pseudomonas aeruginosa: Table 17 Table 18 These results demonstrate at least a short period reduction in bacterial populations on both air-dried and rinsed catheter sections.

Example 6 Altered CBM values when tetra-sodium EDTA is combined with ethanol Solutions having a range of tetra-sodium EDTA concentrations (0, 0.1, 0.5, 1, 2, 3, 4 and 8 mg / ml, p / v) were formulated with water and ethanol (to achieve the final ethanol concentrations of 0, 0.1, 0.5, 1, 5, 10, 20 and 40% in water) to test the efficacy of EDTA solutions alone, alcohol solutions alone , and EDTA / alcohol solutions. Concentrated stock solutions of tetra-sodium EDTA were prepared in distilled water and ethanol was added to the concentrated aqueous stock solutions to provide the appropriate ethanol concentration.

Method: • Cultivate an organism in nutrient broth overnight at 37 ° C. • Alcohol stock solutions and tetra-sodium EDTA are used to fill in a grid configuration in 96-well plates (one per culture), using EDTA solutions that have 0, 0.1, 0.5, 1, 2, 3, 4 and 8 mg / ml concentration of tetra-sodium, w / v, in isopropyl alcohol solvents containing 0, 0.1, 0.5, 1, 5, 10, 20 and 40% alcohol, v / v, in water. • Each well contains 150 ul of each diluent and 50 ul of organism at IxlO8 cfu / ml. • In periods of time of 5 minutes, 6 hours and 24 hours each cavity is cultivated by placing a lid of 96 pins on the plate (and in each cavity) then the lid is transferred to a 96-well plate, which contains 300 ul of new nutrient broth in each cavity. Incubate overnight at 37 ° C. Incubate each inoculum plate at 37 ° C during the incubation period. • Record the turbidity of each cavity after 24 hours. The results for several organisms are shown later. Table 19 The solutions of EDTA tetra-sodium in water were more effective in the extermination of tested microorganisms than in the ethanol solutions (alone). The tetra-sodium EDTA combination in alcohol solutions exterminates the microorganisms tested at lower concentrations. The 2 mg / ml (0.2 w / v) solution of EDTA tetra-sodium in 1% alcohol provides excellent results and has a bactericidal effect on all organisms tested. This antiseptic solution is effective at lower concentrations of tetra-sodium EDTA and ethanol than either tetra-sodium EDTA solutions in water alone or ethanol alone. In addition, it is cost effective, safe and convenient to make and use. The antiseptic compositions of the present invention for topical application thus comprise salts of EDTA in a mixed aqueous solvent and ethanol.

Example 7 Solubility of tetra-sodium EDTA in ethanol and effect on pH The solubility of tetra-sodium EDTA in ethanol was tested, and the pH of various tetra-sodium solutions was measured in alcohol solvents.

Method: • Tetra-sodium EDTA was weighed in duplicates through the 10-100 mg interval in 1.5 ml Eppendorf tubes. One ml of 74% ethanol was added to each tube and vortexed by 3 Os. • To the duplicate set of heavy tetra-sodium EDTA, 0.5 ml of sterile distilled water was added and mixed by vortex, followed by 0.5 ml of 74% ethanol. • Each of the tetra-sodium EDTA tubes was tested for pH where solubility was observed. The experimental results showed that tetra-sodium EDTA were completely insoluble in a 74% ethanol solution. further, the results showed that, when tetra-sodium was dissolved in distilled water in concentrations in the range of 10-100 mg / ml, w / v, tetra-sodium EDTA remains in solution during the addition of ethanol. One such preferred technique involves solubilizing salt (s) of EDTA in a first aqueous solution, and then adding ethanol or another solvent in which the EDTA salts are less soluble or insoluble. Prepared in this way, EDTA salt solutions are expected to be stable over time. The pH values measured for several solutions were as follows: 74% ethanol, only pH 7.8 Water pH 7.1 + 10 mg of EDTA tetra-sodium pH 9.0 + 20 mg of EDTA tetra-sodium pH 10.8 + 40 mg of EDTA tetra-sodium pH 11 + 80 mg of EDTA tetra-sodium pH 11.15 + 100 mg of EDTA tetra-sodium pH 11.25 Example 8 Autoclaving effect at 121 ° C in tetra-sodium EDTA solutions The autoclave effect was tested in solutions of Tetra-sodium EDTA to determine if the autoclave could be used to sterilize tetra-sodium EDTA solutions prior to use. The methodology used and the results are described later.

Method: • Make duplicates of 0, 20, 80 and 100 mg / ml of tetra- sodium EDTA in sterile water and sterile, liquid nutrient agar at 50 ° C. • Leave a set at room temperature (not hot) and autoclave a set (hot). • The next day place all the agar bottles in a steam boiler to melt for 40 minutes.

Measurement of diffusion zones: • Use a perforacorchos, puncture two holes in 16 plates of fresh blood agar. • Make a suspension of 0.5 McFarland of CNS and spread using a sterile swab on the plates to create a sieve. • Pipet 150 ul of each of the tetra- sodium solutions in duplicates by puncturing holes and incubate at 37 ° C overnight. • The next day measure the dissemination areas and record the results. The results, measured in zone sizes (mm), are presented later. The zone sizes of the controls were plotted against the concentration, to allow the determination of real EDTA concentrations in the test samples, which are also presented later. These results demonstrate that the autoclave of tetra-sodium EDTA compositions, whether in sterile water or in agar, does not materially affect the antimicrobial activity of the tetra-sodium EDTA compositions.

Table 20: Zone Sizes in mm Table 21: Actual concentration of EDTA Example 9 Autoclave effect at 121 ° C in different formulations of EDTA The autoclave effect was tested on different formulations of EDTA solutions to determine if the autoclave could be used to sterilize several EDTA solutions prior to use. The methodology used and the results are described later.

Method: Preparation of agar • Place 50 ml of Nutrient agar solution in sterile glass bottles of 7X 100 ml. • Do not add EDTA powder to the first bottle (labeled with 0) • Add 2000 mg of EDTA powder to the second bottle (labeled with 40 mg / ml auto) • Add 4000 mg EDTA powder to the third bottle (labeled with 80 mg / mg auto) • Add 5000 mg of EDTA powder to the fourth bottle (labeled with 100 mg / ml auto) • Do not add EDTA to bottles five, six and seven (but they are labeled with 40, 80 and 100 mg / ml WITHOUT an autoclave), leave at room temperature. • Do this for each EDTA formulation to test, and autoclave all bottles, auto indicator, at 121 ° C for 20 minutes. • The next day, all the bottles are placed in a steam bath to melt the agar to pour. • Once melted, allow to cool to 50 ° C before adding the appropriate amount of EDTA to the bottles labeled No autoclave. All bottles are now ready to be tested.

Measurement of diffusion areas • Use a piercing drill, puncture 2 holes in 7 fresh blood agar plates. • Make a suspension of 0.5 McFarland of CNS and spread using a sterile swab on the plates to create a sieve. • Pipet 150 ul of each bottle into 2 separate "punched holes" and incubate at 37 ° C overnight.

• Do this for each EDTA formulation. • The next day measure the dissemination areas and record results. Duplicate holes are used and 2 measurements are made per zone. Copper and ferric EDTA solutions do not produce some zones. The effect of heat on these solutions, therefore, can not be measured using this method. The zone sizes measured for solutions of di-ammonium EDTA, di-potassium EDTA and magnesium EDTA are given below. The zone sizes of the controls (without heat) were plotted against the concentration to allow the determination of real EDTA concentrations in the test samples (hot), and the results are provided later.

Table 21: Zone sizes (mm) Table 22: Real autoclave EDTA values The results demonstrate that the autoclave does not decrease the effectiveness of most EDTA salt compositions. The autoclave of antiseptic compositions of the present invention can therefore be made by following the preparation to provide the sterile antiseptic compositions.

Example 10 pH values of EDTA salts, calcium chloride and sodium citrate The pH values of various salt solutions of EDTA, calcium chloride and sodium citrate were measured using distilled water as the solvent and at specific concentrations. The results are shown later. 10% acid free EDTA pH 4.7 di-ammonium EDTA 10% pH 4.38 calcium and sodium EDTA 10% pH 6.68 di-potassium EDTA 10% pH 4.5 copper EDTA 10% pH 6.15 tetra-sodium EDTA 10% pH 11.6 EDTA tetra -sodic 2% pH 11 TS neutralized EDTA calcium chloride pH 7.3 Calcium chloride, 1 molar pH 3.8 Sodium citrate 50%, 25% pH 8.5 Example 11 Confirmation of the anticoagulant properties of EDTA solutions The anticoagulant properties of EDTA solutions were verified using the following methodology.

Method: • 100 μl of aliquots from a range of concentrations (0.5-100 mg / ml) of tetra-sodium or disodium EDTA solutions, adjusted to a pH of 11.0-11.6, were placed in plastic-covered tubes. • 900 μl of fresh blood from healthy volunteers were added to each aliquot of EDTA solution and mixed lightly by inverting the blood tubes at regular intervals. The results revealed that the control tubes containing blood without EDTA solution had coagulation times of 10-23 minutes. The tubes containing di-sodium EDTA solutions all had coagulation times in excess of 5 days. The tetra-sodium EDTA tubes having a concentration greater than 1 mg / ml had coagulation times in excess of 5 days. The tetra-sodium EDTA tubes having a concentration of 0.5 mg / ml coagulated in 28 minutes. Therefore, tetra-sodium EDTA is effective as an anticoagulant at concentrations in excess of 1 mg / ml (1% w / v).

Example 12 Osmolarity of Tetra-sodium salt suspensions The osmolarity and lysis of red blood cells of tetra-sodium EDTA solutions in water and physiological saline having various concentrations were tested using standard laboratory techniques. The lysis of the red blood cells was tested by adding 50 ul of EDTA blood in 2 ml of each concentration of each solution for 2 hours. The Plasma Osmolarity interval was 275-295 m / osmol.

Table 23 Example 13 Efficacy of three EDTA Salts in the Artificial Urine Crystals (COA) solution A problem with urinary catheters is that urine crystals tend to accumulate on the surface of the catheter. The deposition of urine crystals can promote microbial colonization and / or the formation of biofilms, as well as reduce the flow through the catheter. Therefore, it may be desirable to use a disinfectant composition in connection with urinary catheters that reduce the formation of urine crystals. The efficacy of three EDTA salt solutions in the dissolution of artificial urine crystals was tested using the methodology described below.

Materials: • Artificial urine in a 25 ml plastic universal container with urease, incubated at 45 ° C for 7 days. • Di-ammonium, di-potassium and tetra-sodium EDTA solutions at 100 mg / ml.

Method: • Centrifuge artificial urine crystals at 4000 rpm for 2 minutes. • Decant the supernatant and wash the crystals in water followed by centrifugation. • Resuspend the crystals to 1 ml in water and aliquot 200 ul in four universal containers. • Add 4 ml of 100 mg / ml solution of each EDTA salt and water as a control to each universal container at room temperature. • After 1, 2 and 3 hours visually, the crystal dissolution was observed compared to the control.

The results are shown later. All EDTA salt solutions reduce the deposit of urine crystals compared to an aqueous solution. Therefore, EDTA salt solutions are suitable for use with urinary catheters.

Table 24 All references cited herein, including patent references and non-patent publications, are hereby incorporated by reference in their entirety. While in the above specification this invention has been described in relation to certain preferred embodiments, and many details have been described for purposes of illustration, it will be apparent to those skilled in the art that the invention is susceptible to additional embodiments and that certain details described in the present they can be varied considerably without departing from the basic principles of the invention. It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Claims

CLAIMS Having described the invention as above, it is claimed as property - what is contained in the following claims:

1. Antiseptic solution, characterized in that it comprises at least one salt of ethylenediamine tetraacetic acid (EDTA) at a concentration of at least 0.01% w / v and at least one solvent, wherein the solution has a pH of at least 8.0 and possesses bacterial activity against a broad spectrum of bacteria.

2. Solution according to claim 1, characterized in that it has a pH between 8.5 and 12.5.

3. Solution according to claim 2, characterized in that it has a pH between 9.5 and 11.5.

4. Solution according to claim 3, characterized in that it has a pH between 10.5 and 11.5.

5. The solution according to claim 1, characterized in that at least one salt of EDTA is selected from the group consisting of: sodium di-EDTA; EDTA tri-sodium; Tetra-sodium EDTA; Ammonium EDTA; Di-ammonium EDTA; Potassium EDTA; DI-potassium EDTA; Cupric di-sodium EDTA; Disodium magnesium EDTA; Ferric sodium EDTA; and combinations thereof.

6. The solution according to claim 5, characterized in that at least one salt of EDTA is selected from the group consisting of: tri-sodium EDTA; Tetra-sodium EDTA; and combinations thereof.

7. The solution according to claim 1, characterized in that at least one EDTA salt is present at a concentration of 0.2% w / v at 10.0% w / v.

8. The solution according to claim 7, characterized in that at least one EDTA salt is present at a concentration of 0.2% w / v at 6.0% w / v.

9. The solution according to claim 8, characterized in that at least one EDTA salt is present at a concentration of 0.2% w / v at 4.0% w / v.

10. Solution in accordance with the claim I, characterized in that the solvent is selected from the group consisting of: water; Saline solution; alcohols; and combinations thereof.

11. The solution according to claim 10, characterized in that the solvent is a combination of water and ethanol.

12. Solution in accordance with the claim II, characterized in that it comprises ethanol in an amount of 0.1% to 10% w / v.

13. The solution according to claim 1, characterized in that it possesses bactericidal activity against bacteria in planktonic forms and in sessile forms.

14. Solution according to claim 1, characterized in that it additionally possesses at least one property selected from the group consisting of: (a) anticoagulant activity; (b) fungicidal activity against fungal pathogens; (c) inhibitory activity against protozoan infections; (d) inhibitory activity against infections of the ibex; (e) safe and biocompatible, at least in modest volumes, in contact with a patient; (f) safe and biocompatible, at least in modest volumes, in the patient's bloodstream; and (g) safe and biocompatible with industrial objects and surfaces.

15. The solution according to claim 1, characterized in that the solution is sterile and free of pyrogens.

16. Dry form of antiseptic solution according to claim 1, characterized in that in reconstitution with a solvent, forms the solution according to claim 1.

17. Formulation of the solution according to claim 1, characterized in that it is suitable for topical application to surfaces and objects.

18. Time release formulation of the solution according to claim 1, characterized in that it provides antiseptic activity for a prolonged period of time.

19. Antiseptic solution, characterized in that it comprises a solvent and at least one salt of sodium EDTA at a concentration of between 0.01% and 15% w / v, where the solution has a pH of at least 9.0 and possesses bactericidal activity against a broad spectrum of bacteria.

20. The solution according to claim 17, characterized in that the solvent is selected from the group consisting of: water; Saline solution; alcohols; and combinations thereof.

21. Antiseptic solution, characterized in that it consists essentially of at least one salt of ethylenediamine tetraacetic acid (EDTA) at a concentration of at least 0.01% w / v and at least one solvent, where the solution has a pH of at least 8.0 and possesses bactericidal activity against a broad spectrum of bacteria.

22. The solution according to claim 21, characterized in that it has a pH between 8.5 and 12.5.

23. Solution in accordance with the claim 21, characterized because it has a pH between 9.5 and 11.5.

24. The solution according to claim 21, characterized in that it has a pH between 10.5 and 11.5.

25. The solution according to claim 21, characterized in that at least one EDTA salt is selected from the group consisting of: sodium di-EDTA, tri-sodium EDTA; Tetra-sodium EDTA; Ammonium EDTA; Di-ammonium EDTA; Potassium EDTA; DI-potassium EDTA; Cupric di-sodium EDTA; EDTA di-sodium magnesium; Ferric sodium EDTA; and combinations thereof.

26. The solution according to claim 25, characterized in that at least one EDTA salt is selected from the group consisting of: tri-sodium EDTA; Tetra-sodium EDTA; and combinations thereof.

27. Solution in accordance with the claim 21, characterized in that at least one EDTA salt is present at a concentration of 0.2% w / v at 10.0% w / v.

28. Solution in accordance with the claim 27, characterized in that at least one EDTA salt is present at a concentration of 0.2% w / v at 6.0% w / v.

29. Solution in accordance with the claim 28, characterized in that at least one EDTA salt is present at a concentration of 0.2% w / v at 4.0% w / v.

30. The solution according to claim 21, characterized in that the solvent is selected from the group consisting of: water; Saline solution; alcohols; and combinations thereof.

31. The solution according to claim 30, characterized in that the solvent is a combination of water and ethanol.

32. Solution according to claim 30, characterized in that it comprises ethanol in an amount of 0.1% to 10% w / v.

33. The solution according to claim 21, characterized in that it possesses bactericidal activity against bacteria in planktonic forms and in sessile forms.

34. Method for inhibiting the growth and proliferation of a population of at least one undesirable microorganism on a surface or within an object, characterized in that it comprises contacting the surface or object with a solution according to any of claims 1-33. .

35. Method according to claim 34, characterized in that the undesirable microorganism is selected from the group consisting of: microbial populations; fungal pathogens; protozoa populations; and amíbicas populations.

36. Method according to claim 35, characterized in that the undesirable microorganism is Acan thamoeba.

37. Method according to claim 34, characterized in that the surface or object is selected from the group consisting of: catheters; ducts and medical tubes; intravascular devices; implanted medical devices; medical and veterinary instruments; contact lenses; optical implants; dental, orthodontic and periodontal devices; water storage, distribution and treatment facilities, - industrial equipment; and food preparation and processing equipment.

38. Method for inhibiting the growth and proliferation of a biofilm, characterized in that it comprises contacting the biofilm with a solution according to any of claims 1-33.

39. Cover for use in wound healing, characterized in that it is impregnated with a solution according to any of claims 1-33.