KR20130116440A - Pharmaceutical composition comprising 5-azacitidine for preventing or treating infectious disease induced by biofilm - Google Patents

Abstract

PURPOSE: A pharmaceutical composition containing 5-azacitidine for preventing or treating infectious diseases caused by a biofilm is provided to suppress proliferation of bacteria and formation of a biofilm and to show superior antibacterial effects. CONSTITUTION: A pharmaceutical composition for preventing or treating infectious diseases caused by a biofilm contains 5-azacitidine as an active ingredient. The infectious diseases are selected among dental caries, periodontitis, otitis media, infection in bones and muscles, necrotizing fasciitis, biliary system infection, bacterial prostatitis, infective endocarditis, cystic fibrosis pneumonia, melioidosis, nosocomial infection, intensive care unit (ECU) pneumonia, urinary catheter cystitis, continuous ambulatory peritoneal dialysis (CAPD) peritonitis, and biliary stent blockage. A coating composition for suppressing the formation of a biofilm contains 5-azacitidine as an active ingredient. The coating composition is coated on a medical material or medical implant.

Description

Pharmaceutical composition comprising 5-azacitidine for preventing or treating infectious disease induced by biofilm}

The present invention relates to a pharmaceutical composition for preventing or treating an infection caused by a biofilm, which contains 5-azacitidine as an active ingredient.

Most bacteria are generally in the form of planktonic form in water, but biofilms that form colonies on the surface of solids when viable conditions are poor due to reduced nutrients or attack by antibiotics. The phenotype changes to form. Biofilm is a collection of microorganisms in which microorganisms such as bacteria, yeast and mold are attached to the surface of human body (skin, mouth, tooth, etc.) or facilities (piping, storage tank, etc.) under specific conditions. It can also be used as a habitat to speed up the growth of bacteria. The microorganisms of these biofilms can cause serious problems in industry and medicine, and research is required.

Biofilm formation occurs through the process of bacteria forming glycoprotein substrates around cells in three dimensions and attaching them to the surrounding solids.By the series of processes called quarum sensing, bacteria lack nutrition. The substance can be confined in a matrix and protected from external attacks (antibiotics, hypoxia). In addition, the formation of biofilm lowers the metabolic rate necessary for the survival of bacteria, enabling survival even with a small amount of nutrients, thereby increasing the viability in poor environments. Biofilms are a common phenomenon in nature, and even in rivers with a constant flow rate, bacteria are known to live on the surface of rocks in the form of biofilms.

However, biofilms cause a lot of clinical problems in human infectious diseases. For example, when bacteria are present in the form of biofilm on the surface of teeth to form tartar, it is known to cause tooth decay and periodontal disease. The most influential factors associated with oral health are the occurrence of tooth decay or tooth decay, gum disease or periodontal disease, and unpleasant bad breath, the root cause of which is caused by pathogens in the oral cavity. It is a biofilm. The pathogens are present in a solution state such as saliva when they are not attached to the surface of the oral cavity and can be easily removed together with gargle or drink intake, so there is little chance of causing diseases in the oral cavity. However, the pathogens are attached to the surface of the oral cavity for effective survival, thereby forming a biofilm. Once the biofilm is formed, the defense against external physical and chemical stimuli is significantly increased, so it is not only difficult to remove by general physical methods but also temporarily removed only by using a highly toxic antimicrobial agent or a large amount of antimicrobial agent. Can be. When left unattended, these biofilms become thicker and stronger, forming calculus and becoming a habitat for numerous bacteria, causing various oral diseases. In addition, when biofilm is present on the surface of the central vein or implant, it is not removed by the usual antibiotics, and bacteria are released from the biofilm, which shows a periodic fever.

Biofilm formation has also been reported on the surface of implants and medical materials such as tracheostomy tubes and cochlear implants. Until now, the treatment of such infection with biofilm is known to be the only solution to remove the implants or physically remove the biofilm habitat.

On the other hand, 5-azacitidine is a substance used as an anticancer agent for treating myelodysplastic syndrome, and is inserted into DNA during cell proliferation or RNA during transcription to be a DNA methyl transferase ( inhibits methyltransferase, reducing DNA methylation. By this mechanism, by inhibiting the gene expression of the cell can play a role in blocking the proliferation of cancer cells. However, there are no studies showing that 5-azacytidine has a prophylactic or therapeutic effect on biofilm-induced infections.

The inventors of the present invention continued to study new substances for treating infection caused by biofilm, and confirmed that 5-azacytidine had an effect of inhibiting bacterial growth and biofilm formation. Was completed.

An object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of infection caused by a biofilm containing 5-azacitidine as an active ingredient.

Still another object of the present invention is to provide a coating composition for inhibiting biofilm formation containing 5-azacytidine as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating infection caused by a biofilm containing 5-azacytidine as an active ingredient.

The present invention also provides a coating composition for inhibiting biofilm formation using 5-azacytidine as an active ingredient.

5-azacitidine of the present invention is excellent in antibacterial effect by inhibiting the growth of bacteria and the formation of biofilm, it can be usefully used for the prevention or treatment of infection caused by biofilm.

1 is a diagram observing the effect of 5-azacytidine on the formation of biofilm under an electron microscope after culturing pneumococci in the medium and inducing the formation of biofilm (biofilm).

Hereinafter, the present invention will be described in detail.

The present invention provides a pharmaceutical composition for the prevention or treatment of infection caused by a biofilm containing 5-azacitidine as an active ingredient.

5-azacytidine of the present invention is excellent in antibacterial effect by reducing the gene expression of Streptococcus pneumonia by inhibiting the formation of biofilm. Therefore, 5-azacytidine can be usefully used for the prevention or treatment of infection caused by biofilm.

The infectious disease caused by the biofilm is preferably an infectious disease that can be suppressed by an antimicrobial effect, and this includes an infection that has been treated with conventional disinfectants and antibiotics and an infectious disease when the disease is diagnosed.

Examples of the infectious diseases caused by the biofilm are not limited thereto, but are not limited to tooth decay, periodontitis, otitis media, musculoskeletal infection, necrotizing fasciitis, cholangitis infection, osteomyelitis, bacterial prostatitis, natural valve endocarditis, cystic fibrosis pneumonia, Meloidosis, and nosocomial infections such as intensive care unit (ICU) pneumonia, urinary catheter cystitis, peritoneal dialysis (CAPD) peritonitis, and biliary stent blockage do. Biofilm formation includes sutures, extractions, arteriovenous areas, scleral buckle sites, contact lenses, IUDs, tracheal tubes, Hickman catheter, central venous catheter, artificial heart valve, valve graft, orthopedic This may affect the device and penis implants. Other applications are described in Costerton J et al and Costerton J and Steward (2001 Battling Biofilms, Scientific American pp 75-81), which is incorporated herein by reference. Other areas where biofilms can form include cavities that can progress to gum disease and cavity, contact lenses that can progress to eye infections, ears and pneumonia that can lead to chronic infections. Contains lungs that can The infection may be cystic fibrosis. Infections can result from skin infections, burn infections, and / or wound infections. The compositions of the present invention may be particularly suitable for treating infections in immunocompromised individuals.

The biofilm is caused and formed by the pathogen.

The pathogen as used herein refers to a microorganism causing a disease such as, but not limited to, bacteria or viruses, and refers to a causative agent of various infectious diseases such as intestinal infection, respiratory infection, and urinary tract infection. Examples include Salmonella spp., Shigella spp., Vibrio parahaemolyticus, Vibrio choreae, Escherichia coli O-157, Campylobacter jejuni, stomach Clostridium difficile, Clostridium perfringens, Elsinia enterocolitica, Helicobacter pylori, Amoeba erythrocytes, Entemoeba histolytica, Bacillusu cereus, Botulintrius botulin ), Haemophilus influenzae, Streptococcus pneumoniae, Chlamydia pneumoniae, Legionella pneumoniae, Branhamella pneumoniae, Mycobacterium tuberculosis Mycobacterium tuberculosis , Type A Lactobacillus (Storeptcoccus pyogenes), Diphtheria bacteria (Corynebacterium diphtheriae), Pertussis (Bordetella pertussis), Scabies (Chramidia psittaci), Pseudomonas aeruginosa ( Pseudo monas aerginosa, methicillin resistant Staphylococcus aureus (MRSA), Escherichia coli, Klebsiella pneumoniae, Enterobacter spp., Proteus spp. Examples include Acinetobacter spp., Enterococcus faecalis, Staphylococcus saprophyticus, and B-type Lactobacillus (Storeptcoccus agalactiae).

In the present invention, disinfection refers to killing pathogens, and it does not matter whether non-pathogenic microorganisms remain.

The disinfectants include, but are not limited to, hydrogen peroxide, chlorine, iodine, phenols, alcohols, chlorohexidine gluconate, triclosan, and the like.

When the disease is diagnosed, the diagnosis may be performed using a medical device or the like, but a medical device having a surface of a polymer material or a metal material is not limited thereto.

The composition of the present invention may contain one or more known active ingredients having a prophylactic or therapeutic effect of an infection caused by a biofilm together with 5-azacitidine.

The composition of the present invention can be prepared by including one or more pharmaceutically acceptable carriers in addition to the above-described active ingredients for administration. Pharmaceutically acceptable carriers may be used in combination with saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol and one or more of these components, if necessary, as antioxidants, buffers And other conventional additives such as bacteriostatic agents can be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Further, it can be suitably formulated according to each disease or ingredient, using appropriate methods in the art or by the method disclosed in Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA.

The composition of the present invention can be administered orally or parenterally according to the desired method, the dosage is depending on the weight, age, sex, health condition, diet, time of administration, administration method, excretion rate and severity of disease, etc. of the individual. The range varies. The daily dose of 5-azacytidine is 0.5-3.0 mg / kg, preferably about 1.0-2.0 mg / kg, but can be added or subtracted according to clinical results, preferably administered once or several times a day.

The composition of the present invention may be used alone or in combination with existing disinfectants or antibiotics, hormone therapy, drug treatment and biological response modifiers for the prevention or therapeutic effect of infection caused by biofilm.

The present invention also provides a coating composition for inhibiting biofilm formation using 5-azacytidine as an active ingredient.

The coating agent is characterized in that the coating on the medical device, medical material, medical implant.

The medical device is not limited to this, but consultation and diagnosis equipment, clinical examination equipment, radiation equipment, surgery related equipment and equipment, etc. ), Treatment Apparatus & Equipment, Rehabilitation Medicine, Physiotherapy Appartus, Ophthalmic Appartus, Dental Appartus, Central Supply Equipment, Hospital Equipment Examples include Hospital Accmmodation & Emergency Equipment, Oriental Medicine & Equipment, Pharmaceutical Equipment, Obesity Cure & Healthcare Equipment, Medical Supplies and Consumables. Can be.

The medical material is classified into a metal material, an inorganic material, a polymer material, a composite material and the like according to the material. Metal materials are divided into metal materials such as cobalt, chromium, stainless steel, touch carbon and decarburization. Inorganic materials include bioglass and ceramic, and polymer materials include natural polymer, bioplastic, Synthetic polymer materials, synthetic rubber, synthetic fibers, and the like, and medical composite materials include, but are not limited to, Fiber Reinforced Plastics, glass fibers, and carbon fibers.

Medical implants range from very simple, such as catheter, to very complex and sophisticated devices, such as artificial organs or sensors. However, medical implants that are implanted under the patient's subcutaneous body to cure ailments or discomfort can be life threatening. This is because serious infections can occur during the implantation of medical implants into patients. More than half of hospital infectious diseases are associated with the implantation of medical devices, examples of which include intravascular implants, stents, heart stents, peripheral stents, orthopedic implants, bone or joint implants, artificial heart, artificial heart valves, Subcutaneous and / or intramuscular implants are, but are not limited to these.

The surface of the medical device is preferably a polymer material or a metal material.

The polymer material refers to a polymer material used in medical activities, that is, a polymer used as an artificial organ to replace a medical device or a part of a body used in various medical activities including diagnosis and treatment. Areas of use include transfusion equipment, including transfusion bags, syringes, soft contact lenses, kidney filters, urethra, heart valves, artificial blood vessels, bones, skin, tendons, ears, jaw, eyes, teeth and tooth fillers Etc. Materials used include, but are not limited to, polyvinyl chloride, polyethylene, natural rubber, silicone rubber, polypropylene, polyurethane, and the like.

The metal material includes, but is not limited to, stainless steel, cobalt alloy, titanium alloy, metal biomaterial, cobalt chromium alloy, titanium alloy, tantalum, shape memory alloys, and the like.

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples.

Example 1 Effect of 5-Azacytidine on Biofilm Formation

In order to confirm the effect of 5-azacitidine on the biofilm of the present invention, the following experiment was performed.

Specifically, Streptococcus pneumonia was incubated in a medium (Triptic soy broth, TSB) to induce the formation of biofilms. Here, 5-azacytidine was treated, and expression of genes in pneumococci was observed using a polymerase chain reaction (PCR).

The results are shown in Table 1.

Gene Expression Changes of Streptococcus pneumonia by 5-Azacytidine gene Average Standard error T test LuxS 0.084 0.014142 0.014129 metE 0.789 0.005657 0.001157 metK 0.3355 0.055928 0.103945 cmK 0.298 0.014422 0.019092 mraW 0.5795 0.00137 0.161927 mutL 0.6075 0.084668 0.161927 Pfs 0.429 0.017647 0.066468

As shown in Table 1, it was confirmed that the pneumococcal gene was reduced by treatment with 5-azacytidine.

Example 2 Inhibitory Effect of Biofilm Formation with 5-Azacytidine Concentration

Streptococcus pneumonia was incubated in medium (Triptic soy broth, TSB) to induce the formation of biofilms. Here, 5-azacytidine was treated at various concentrations (0 uM, 100 uM, 500 uM), and changes in biofilm formation were observed through electron microscopy. The results are shown in Fig.

As shown in FIG. 1, when 5-azacytidine was not treated, a biofilm was formed, and when 5-azacytidine was treated, formation of biofilm was suppressed.

Through the above results, it can be seen that 5-azacytidine inhibits the formation of the biofilm.

Examples of formulations for the composition of the present invention are illustrated below.

Formulation Example 1 Preparation of Powder

5-Azacytidine 200mg

Lactose 100mg

The above ingredients were mixed and filled in airtight cloth to prepare a powder.

Formulation Example 2 Preparation of Tablet

5-Azacytidine 200mg

Corn Starch 100mg

Lactose 100mg

2 mg magnesium stearate

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

Formulation Example 3 Preparation of Capsule

5-Azacytidine 200mg

Corn Starch 100mg

Lactose 100mg

2 mg magnesium stearate

After mixing the above components, the capsule was prepared by filling in gelatin capsules according to the conventional method for producing a capsule.

Formulation Example 4 Preparation of Injection

5-Azacytidine 200mg

Mannitol 100mg

Na2HPO4, 12H2O 2mg

Sterile sterilized water for injection

Injectables were prepared by mixing the above ingredients per ampoule (2 ml) according to the usual method for preparing injectables.

Claims (5)

A pharmaceutical composition for the prevention or treatment of infection caused by a biofilm containing 5-azacytidine as an active ingredient. According to claim 1, The infection caused by the biofilm is caries, periodontitis, otitis media, musculoskeletal infection, necrotizing fasciitis, cholangitis infection, osteomyelitis, bacterial prostatitis, natural valve endocarditis, cystic fibrosis pneumonia, melo Selected from the group consisting of meloidosis, nosocomial infection, intensive care unit (ICU) pneumonia, urinary catheter cystitis, peritoneal dialysis (CAPD) peritonitis and biliary stent blockage A pharmaceutical composition for the prevention or treatment of infections caused by biofilm, characterized in that at least one. The method of claim 2, wherein the biofilm is Salmonella spp., Shigella spp., Vibrio parahaemolyticus, Vibrio choreae, Escherichia coli O-157 Escherichia coli O-157 , Campylobacter jejuni, Clostridium difficile, Clostridium perfringens, Elsinia enterocolitica, Helicobacter pylori, Amobacterium bacterium Entemoeba histolytica, Bacilleus cereus, Botulinum botulinum, Haemophilus influenzae, Streptococcus pneumoniae, Chlamydia pneumoniae, Legionella pneumoniae, Branharella bacterium tuberculosis ), Mycoplasma pneumoniae, type A lytic bacteria (Storeptcoccus pyogenes), diphtheria bacteria (Corynebacterium diphtheriae), pertussis (Bordetella pertussis), scabies (Chramidia psittaci), Pseudo monas aerginosa, methicillin resistant Staphylococcus aureus (MRSA), Escherichia coli, Klebsiella pneumoniae, Enterobacter sp. Proteus spp., Acinetobacter spp., Enterococcus faecalis, Staphylococcus saprophyticus, Form B (Storeptcoccus agalactiae) formed by one or more species selected from the group consisting of A pharmaceutical composition for preventing or treating infection caused by biofilm. Coating composition for inhibiting biofilm formation containing 5-azacytidine as an active ingredient. The coating composition of claim 4, wherein the coating composition is coated on a medical device, a medical material, or a medical implant.

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