KR101349752B1 - Novel antimicrobial comoposition having quorum sensing inhibiting activity and antimicrobial activity - Google Patents

Abstract

The present invention relates to an antimicrobial composition containing a specific compound or salt thereof as an active ingredient, wherein the compound or salt thereof is coated on an artificial insert, for example, a central vein tube, an artificial joint, or an artificial valve, in addition to an antimicrobial agent or health food. By suppressing the biofilm formation of the prosthesis and at the same time can exhibit an antimicrobial activity that directly inhibits bacteria can be very useful as a coating composition for preventing prosthesis infection.

Description

Novel antimicrobial comoposition having quorum sensing inhibiting activity and antimicrobial activity

The present invention relates to a novel antimicrobial composition having a quorum detection inhibitory activity and an antimicrobial activity.

Bacteria invade the host and multiply in a tissue suitable for survival, causing the disease. In this process, signaling materials that control individual concentrations between cells are required. The small molecule that serves as a signal transduction is called a quorum sensing material. When the concentration outside the cell reaches a certain level, it enters the cell and participates in various regulatory functions such as the expression of a protein as a ligand.

Recent studies have attempted to utilize quorum sensing materials as therapeutics and biofilm inhibitors for resistant bacteria.

In particular, the use of various prostheses is increasing in modern medicine. Infections with biofilm formation are common, and in most cases, removal of the prosthesis can only cure the infection. Therefore, not only the pain of patients but also the economic loss is very large.

Meanwhile, Korean Patent Publication No. 2004-0099280 discloses effects of 4,5-dihydroxy-2-cyclopenten-l-one (DHCP) on the gene expression and quorum detection of bacteria. 0777780 discloses an antimicrobial composition that inhibits quorum detection.

However, the development of a material capable of effectively inhibiting biofilm is required for derivatives of quorum detection inhibitors, and if such a material is developed, it can alleviate the pain of patients and have a great economic and industrial effect.

In particular, implants such as central venous tubes, prosthetic joints, prosthetic valves, and cardiac pacemakers, although difficult to treat and are the main cause of antibiotic resistance, can effectively control biofilms on the implant surface. No effective substance has been developed.

Accordingly, the present inventors completed the present invention by selecting a substance showing excellent antimicrobial activity against Gram-positive bacteria and multidrug-resistant Gram-positive bacteria with respect to derivatives of the quorum detection inhibitor.

Accordingly, an object of the present invention is to provide a new antimicrobial composition that can effectively control the biofilm having a quorum detection inhibitory activity, as well as having an antimicrobial activity.

In order to achieve the above object, the present invention provides a composition for antimicrobial containing a compound represented by the following formula (1) or a salt thereof as an active ingredient:

[Formula 1]

The compound represented by Formula 1 or a salt thereof may exhibit antimicrobial activity among Gram-positive bacteria or multidrug-resistant Gram-positive bacteria, and in particular, the multidrug-resistant Gram-positive bacteria may be Staphylococcus aureus or Vancomycin-resistant enterococci ( Enterococcus faecium).

The composition may be any one of a pharmaceutical composition or health food.

In particular, the composition is preferably coated on the surface of the prosthesis to prevent infection of the prosthesis. The prosthesis may include a central vein tube, an artificial joint, an artificial valve, an artificial heart pacemaker, and the like.

Compounds or salts thereof according to the present invention exhibit excellent antimicrobial activity against multidrug-resistant Gram-positive bacteria in addition to the quorum-sensitive inhibitory effect, the composition containing the compound or salts thereof may be used as an antimicrobial agent or health food. In addition, by coating the compound or a salt thereof on the prosthesis, for example, a central vein, an artificial joint, or a prosthesis, the prosthetic infection may be inhibited by inhibiting biofilm formation of the prosthesis and at the same time directly inhibiting bacteria. It can be very usefully used as a coating composition for prevention.

 The terms, techniques, and the like described in the present invention are used in the meaning commonly used in the technical field to which the present invention belongs, unless otherwise specified. In addition, all documents referred to in this specification are included in the present specification as a document for explaining the present invention.

Hereinafter, the present invention will be described in more detail.

To prepare a compound according to the invention, first, indole, 2,4-dichloro-benzenethiol is dissolved in a solution containing methanol and water in a volume ratio of 3: 2 and KI and I ₂ are added, followed by N ₂ (g ) And stirred. Thereafter, the solvent is removed, extracted with dichloromethane, dried, filtered, the solvent is removed and separated by a silica gel column to obtain a compound of the present invention.

In addition, the compounds according to the invention can be prepared with the corresponding salts by known methods. Such salts are preferably water-soluble and non-toxic. Preferred salts include salts of alkali metals such as potassium, sodium and the like; Salts of alkaline earth metals such as calcium, magnesium, and the like; And amines such as tetramethylammonium, triethylamine, methylamine, dimethylamine, cyclopentylamine, benzylamine, phenethylamine, piperidine, monoethanolamine, diethanolamine, tris (hydroxymethyl) amine, lysine, arginine, And salts of pharmaceutically acceptable amines such as N-methyl-D-glucamine and the like.

The compounds according to the invention can be prepared with the corresponding acid addition salts by known methods. Such acid addition salts are preferably water soluble with no toxicity. Preferred acid addition salts include inorganic acid salts such as hydrochloride, hydrobromide, hydroiodide, sulfate, phosphate and nitrate, or acetates, lactates, tartarates, oxalates, fumarates, maleates, citrates, benzoates, methane Organic acid salts such as sulfonate, ethanesulfonate, benzenesulfonate, toluenesulfonate, isethionate, glucuronate and gluconate.

The compounds according to the invention or their salts can be prepared as hydrates by known methods.

In particular, the compound according to the present invention or salt thereof exhibits excellent antimicrobial activity against multidrug resistant Gram-positive bacteria in addition to quorum-inhibiting effect.

The composition of the present invention can be used as a pharmaceutical composition or health food.

The pharmaceutical compositions may further comprise suitable carriers, excipients or diluents conventionally used in the manufacture of pharmaceutical compositions.

Examples of the carrier, excipient or diluent which can be used in the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil.

The pharmaceutical composition according to the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions according to a conventional method .

In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose sucrose), lactose, gelatin, and the like.

In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included .

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of suppository bases include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like.

The compound according to the present invention or a salt thereof may be included in an amount of 0.01 to 30.0 parts by weight based on 100 parts by weight of the pharmaceutical composition. When the content is less than 0.01% by weight, the antimicrobial activity is insignificant, and when the content is more than 30.0% by weight, there may be a problem that the antimicrobial activity is insignificant depending on the amount used.

The amount of the compound or salt thereof, which is an active ingredient of the pharmaceutical composition according to the present invention, may vary depending on the age, sex, weight, and disease of the patient, but is 0.001 to 100 mg / kg, preferably 0.01 to 10 mg / kg per day. It may be administered once to several times.

In addition, the dosage of a compound according to the present invention or a salt thereof may be increased or decreased depending on the route of administration, the severity of the disease, sex, weight, age, and the like. Thus, the dosage amounts are not intended to limit the scope of the invention in any manner.

The pharmaceutical composition may be administered to mammals such as rats, mice, livestock, humans, and the like in a variety of routes. All modes of administration can be expected, for example, may be administered by oral, rectal or intravenous, intramuscular, subcutaneous, intrabronchial inhalation, intrauterine dural or intracerebroventricular injection and applied to the prosthesis. It can also be inserted into the human body.

Compounds or salts thereof according to the present invention are 50% lethal dose (LC ₅₀ ) of 2 g / kg or more, the stability is ensured, can be used in the pharmaceutical composition of the present invention.

In addition, the health food is used with other food or food additives in addition to the compound according to the present invention or salts thereof, and may be appropriately used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to its use purpose, for example, prevention, health or therapeutic treatment.

An effective dose of the compound according to the present invention or salt thereof contained in the health food may be used in accordance with the effective dose of the pharmaceutical composition, but for long term intake for health and hygiene purposes or for health control purposes, It may be less than the above range, it is clear that the active ingredient can be used in an amount above the above range because there is no problem in terms of safety.

There is no particular limitation on the type of the health food, and examples thereof include meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, Drinks, alcoholic beverages and vitamin complexes.

Hereinafter, the present invention will be described in detail with reference to the following examples and experimental examples. However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the present invention is not limited to the following Examples and Experimental Examples.

< Example  1>

Indole (100 mg, 0.853 mmol) and 2,4-dichloro-benzenethiol (183 mg, 1.024 mmol, 1.2eq) were dissolved in 30 mL of a solution containing MeOH and water in a volume ratio of 3: 2, and KI (425 mg, 2.56 mmol, 3eq). ), I ₂ (216 mg, 1.71 mmol, 1 eq) was added, followed by substitution with N ₂ (g) and stirring at 24 ° C. for 7 hours. The solvent was removed, extracted with dichloromethane, dried over MgSO ₄ , filtered, the solvent was removed and separated by a silica gel column to give a white solid compound (yield: 75%).

¹ H NMR (500 MHz, Chloroform-d) ppm 6.58 (d, J = 8.3 Hz, 1H), 6.90 (dd, J = 8.3, 2.2 Hz, 1H), 7.17-7.22 (m, 1H), 7.31 (t, J = 7.7 Hz, 1H), 7.35 (d, J = 1.7 Hz, 1H), 7.48 (d, J = 8.3 Hz, 1H), 7.52 (d = 2.8 Hz, 1H), 7.56 (d, J = 7.7 Hz , 1H), 8.51 (br. S., 1H)

¹³ C NMR (126 MHz, Chloroform-d) ppm 99.79, 110.72, 118.38, 120.22, 122.35, 126.10, 126.15, 127.64, 127.90, 129.35, 129.46, 130.23, 135.53, 136.26.

<Experimental Example 1> in vitro  Antimicrobial Activity Review

Clinical and Laboratory Standards Institute (2009) using the compounds synthesized in Example 1. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically. Wayne, PA, Approved standard M7-A8. Clinical and Laboratory Standards Institute The minimum inhibitory concentraion (MIC) of vancomycin-resistant Staphylococcus arueus and vancomycin-resistant Enterococcus faecium was measured in vitro using the standard agar dilution method described in the Table.

As a result, the compound of Example 1 showed excellent antimicrobial activity as shown in Table 1 below.

Target strain MIC ^* (μg / mL) Staphylococcus aureus ATCC29213 (standard strain) One Vancomycin resistance
Staphylococcus aureus Mu50
Standard strain of vancomycin intermediate susceptible Staphylococcus aureus (VISA) 0.5 AMC11094
Clinical strain of VISA isolated from Asan Medical Center One Mu3
Standard strains of different resistance vancomycin intermediate susceptible Staphylococcus aureus (hVISA) One hVISA No. 805
HVISA Clinical Strain Isolated from Asan Medical Center One hVISA No. 609
HVISA Clinical Strain Isolated from Asan Medical Center One hVISA No. 722
HVISA Clinical Strain Isolated from Asan Medical Center One MRSA No. 379
Vancomycin Insensitive Strains Isolated from Asan Medical Center
(Vancomycin MIC = 3 μg / mL) One MRSA No. 495
Vancomycin Insensitive Strains Isolated from Asan Medical Center
(Vancomycin MIC = 3 μg / mL) One MRSA No. 582
Vancomycin Insensitive Strains Isolated from Asan Medical Center
(Vancomycin MIC = 3 μg / mL) One Enterococcus faecium ATCC29212 (standard strain) 2 Vancomycin-Resistant Enterococci Vancomycin Resistant Enterococci No. 5807 One Vancomycin Resistant Enterococci No. 5809 One

< Experimental Example  2> toxicity test

In male Balb / c mice, the compound of Example 1 was suspended in 0.5% methylcellulose solution, respectively, and administered once orally at a dose of 0.5 g / kg, 1 g / kg, and 2 g / kg. Investigate.

After this administration, we observed the mortality of the animal, clinical symptoms, weight change, hematologic test and blood biochemical test, and autopsied the visceral organs and thoracic organs were observed.

As a result, no clinical symptoms or dead animals were found in all animals, and no toxic changes were observed in weight changes, blood tests, blood biochemical tests, and autopsy findings.

As a result, the compounds of the present invention did not show toxic changes up to 2 g / kg in mice, and thus, it was judged that the oral LD50 was 2 g / kg or more.

Hereinafter, the preparation examples of the composition comprising the compound of Example 1 according to the present invention will be described, but the present invention is not intended to be limited thereto but only to be described in detail.

Preparation Example 1 Preparation of Powder

5 mg of the compound of Example 1, 100 mg of lactose and 10 mg of talc were mixed and filled into an airtight cloth to prepare a powder.

&Lt; Formulation Example 2 > Preparation of tablet

Tablets were prepared by mixing 5 mg of the compound of Example 1, 100 mg of corn starch, 100 mg of lactose, and 2 mg of magnesium stearate, followed by compression according to a conventional method for preparing tablets.

Preparation Example 3 Preparation of Capsule

After mixing 5 mg of the compound of Example 1, 100 mg of corn starch, 100 mg of lactose and 2 mg of magnesium stearate, the above ingredients were mixed and filled into gelatin capsules according to a conventional capsule preparation method to prepare a capsule.

Preparation Example 4 Preparation of Injection

After mixing 2 mg of the compound of Example 1 27, a sterile distilled water for injection and a pH adjusting agent, the compound content was prepared per ampoule (2 ml) according to the conventional method for preparing an injection.

&Lt; Formulation Example 5 > Preparation of beverage

The composition of the compound of Example 1 0.05mg, honey 522mg, chitosanamide 5mg, nicotinic acidamide 10mg, riboflavin sodium 3mg, pyridoxine hydrochloride 2mg, inositol 30mg, orthoic acid 50mg and 200ml water and The beverage was prepared using conventional methods by content.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. something to do. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (6)

An antimicrobial composition having a quorum-sensing inhibitory activity and an antimicrobial activity, containing the compound represented by the following Formula 1 or a salt thereof as an active ingredient:
[Chemical Formula 1]

The antimicrobial composition of claim 1, wherein the compound represented by Chemical Formula 1 or a salt thereof exhibits antimicrobial activity among Gram-positive bacteria or multidrug-resistant Gram-positive bacteria. The antimicrobial composition according to claim 2, wherein the multidrug-resistant Gram-positive bacteria is any one of vancomycin-resistant Staphylococcus aureus or Vancomycin-resistant Enterococcus faecium. . The antimicrobial composition of claim 1, wherein the composition is any one of a pharmaceutical composition or a health food. The antimicrobial composition of claim 1, wherein the composition is coated on the surface of the prosthesis to prevent prosthesis infection. The antimicrobial composition of claim 5, wherein the prosthesis is selected from the group consisting of a central venous tube, an artificial joint, an artificial valve, and an artificial heart pacemaker.