KR20190130983A - Novel brominated furanone derivatives, preparation method thereof, and pharmaceutical composition for use in preventing or treating periodontitis containing the same as an active ingredient - Google Patents

Abstract

The present invention relates to a novel brominated furanone derivative, a method for manufacturing the same, and a pharmaceutical composition containing the same as an active component, wherein the novel brominated furanone derivative or the pharmaceutically acceptable salt thereof according to the present invention exhibits a quorum sensing inhibitory activity of bacteria and also can effectively inhibit the formation of a biofilm of a bacteria and bacterial growth, thereby being able to be used as a pharmaceutical composition, a functional health food composition, an antibiotic composition, a toothpaste composition or a mouthwash composition containing the same as the active component. Therefore, it is possible to exhibit useful effects on oral diseases or inflammatory diseases, for example periodontal diseases and the like such as gingivitis and periodontitis.

Description

Novel brominated furanone derivatives, preparation method conjugate, and pharmaceutical composition for use in preventing or treating periodontitis containing the same as an active ingredient

The present invention relates to a novel brominated furanone derivative, a preparation method thereof and a pharmaceutical composition containing the same as an active ingredient.

When a bacterium reaches a certain level of cell concentration, it uses its chemical language to transmit signals between cells and regulate the expression of specific genes. Gram-negative and gram-positive bacteria have a quorum sensing (QS) mechanism, so that gene expression depends on the density of the cell. The quorum sensing can be expressed by proper density recognition, and each bacterial entity accumulates low molecular signaling materials such as autoinducers or pheromones outside the cell to induce active proliferation and quorum. Refers to a series of biological phenomena that fill

Specifically, N-acyl homoserine lactone synthase protein (N-acyl homoserine lactone synthase protein) among the protein constituting the bacteria synthesizes a signaling material called normal acyl homoserine Lactone (AHL). The synthesized normal acyl homoserine lactone freely diffuses through the cell membrane during bacterial growth and accumulates in the environment outside the cell. When the density of bacteria increases and the signaling material accumulated outside the cell reaches a certain concentration, the signaling material enters the cell again and binds to a transcriptional regulator to promote expression of a specific gene.

Bacteria control various physiological phenomena such as formation of biofilm, virulence, bioluminescence, production of antibiotics or delivery of tumor-induced plasmid by conjugation through the quorum sensing mechanism described above. do. For example, in a series of bacterial infections, bacteria first find their way into the host and settle in a habitat suitable for survival. It then survives, neutralizing the host's primary defense system. Finally, bacteria multiply and spread offspring to other hosts. In this process, bacteria express various virulence factors by quorum sensing mechanism.

Most of the antimicrobial agents developed in the past showed antimicrobial performance by killing bacteria, but in controlling the bacteria causing the onset, not only killing the bacteria but also disturbing the communication of the bacteria and delayed breeding of the bacteria. It is also important to prevent it.

A representative function of the bacteria communicating is the biofilm, which stays in the human organs and causes a number of conditions. Examples of conditions or onset include dental caries, gingivitis, periodontitis, otitis media, voice prostheses, hydrocephalus hunts, and cystic fibrosis. Valvular endocarditis, prosthetic heart valves, central venous catheter, prosthetic hip joint, prosthetic knee joint, chronic bacterial prostatitis, Intrauterine devices, urinary catheter, and the like. Therefore, in addition to killing the bacteria, it is required to develop a multi-functional antimicrobial agent that inhibits the function of the biofilm (biofilm) formation of the bacteria in advance by interfering with the communication between the bacteria.

In particular, gram negative bacteria are known to communicate by using a chemical called normal acyl homoserine lactone (NHL). If an antagonist with a structure similar to normal acyl homoserine lactone AHL is synthesized and acted around the bacteria, the gene expression of the bacteria can be prevented to prevent reproduction. Therefore, there is a need for the development of antimicrobial agents having various antimicrobial properties that have a molecular structure that kills microorganisms and at the same time prevents reproduction by interfering with communication between microorganisms.

Patent Document 1 discloses a method for removing microorganisms using a compound having antibacterial activity and functioning as a quorum sensing antagonist. However, the patent document disclosed in the patent document has a low curum sensation inhibitory effect and antibacterial effect. The material has not been developed.

Therefore, while the present inventors are studying a quorum sensing inhibitor, the novel brominated furanone derivative of the present invention or a pharmaceutically acceptable salt thereof exhibits the effect of inhibiting the cure of bacteria and effectively inhibits the biofilm formation of the bacteria. And, it was confirmed that the antibacterial effect was completed the present invention.

Korea Patent Publication 10-2008-0046434

It is an object of the present invention to provide novel brominated furanone derivatives, stereoisomers thereof or pharmaceutically acceptable salts thereof.

Another object of the present invention is to provide a method for preparing the brominated furanone derivative.

Another object of the present invention to provide a pharmaceutical composition for the prevention or treatment of oral diseases containing the brominated puranone derivatives, stereoisomers thereof or pharmaceutically acceptable salts thereof as an active ingredient.

Another object of the present invention is to provide a dentifrice composition for the prevention or improvement of oral disease, containing the brominated puranone derivatives, stereoisomers thereof or pharmaceutically acceptable salts thereof as an active ingredient.

Still another object of the present invention is to provide a goggle composition for the prevention or improvement of oral diseases, containing the brominated furanone derivatives, stereoisomers thereof or pharmaceutically acceptable salts thereof as an active ingredient.

Still another object of the present invention is to provide a nutraceutical composition for the prevention or improvement of oral diseases containing the brominated furanone derivatives, stereoisomers thereof or pharmaceutically acceptable salts thereof as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of inflammatory diseases containing the brominated furanone derivatives, stereoisomers thereof or pharmaceutically acceptable salts thereof as an active ingredient.

Still another object of the present invention is to provide an antibiotic composition containing the brominated furanone derivative, its stereoisomer, or a pharmaceutically acceptable salt thereof as an active ingredient.

In order to achieve the above object,

The present invention provides a compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[Formula 1]

(In the above formula 1,

X is O, S or NR ¹ ,

Wherein R ¹ is unsubstituted or substituted C _1-10 straight or branched chain alkyl, unsubstituted or substituted C _1-10 straight or branched chain alkoxy, unsubstituted or substituted C _3-7 cycloalkyl , 3 to 7 cyclic unsubstituted or substituted heterocycloalkyl, unsubstituted or substituted C _6-10 aryl containing one or more hetero atoms selected from the group consisting of N, O and S, or N, O And 5 to 10 each ring heteroaryl including one or more hetero atoms selected from the group consisting of S,

Wherein the substituted alkyl, substituted alkoxy, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl or substituted heteroaryl are hydroxy, halogen, -CN, -NO ₂ , C _1-5 May be substituted with straight or branched chain alkyl, C _1-5 straight or branched chain alkoxy;

는 비치환 또는 치환된 C_3-7의 사이클로알킬, 하나 이상의 이중결합을 포함하는 비치환 또는 치환된 C_3-7의 사이클로알케닐, N, O 및 S로 이루어진 군으로부터 선택되는 1종 이상의 헤테로 원자를 포함하는 3 내지 7각환의 비치환 또는 치환된 헤테로사이클로알킬, 비치환 또는 치환된 C_6-10의 아릴, 또는 N, O 및 S로 이루어진 군으로부터 선택되는 1종 이상의 헤테로 원자를 포함하는 5 내지 10각환의 헤테로아릴이되,

Is unsubstituted or substituted C _3-7 cycloalkyl, at least one double bond alkenyl cycloalkyl unsubstituted or substituted C _3-7 Al, including, N, O, and one or more heteroatoms selected from the group consisting of S 3 to 7-membered unsubstituted or substituted heterocycloalkyl containing atoms, unsubstituted or substituted C _6-10 aryl, or at least one hetero atom selected from the group consisting of N, O and S 5 to 10 each ring heteroaryl,

Wherein the substituted cycloalkyl, substituted cycloalkenyl, substituted heterocycloalkyl, substituted aryl, or substituted heteroaryl is a straight chain of hydroxy, halogen, -CN, -NO ₂ , C _1-10 Or branched alkyl, C _1-10 straight or branched alkoxy; And

One of W and Y is Br and the other comprises H, unsubstituted or substituted C _6-10 aryl, or a heteroaryl of one member selected from the group consisting of N, O, and S Ring or substituted 5-10 atoms of heteroaryl,

Wherein said substituted aryl, or substituted heteroaryl, is OH, halogen, cyano, nitro, amino, unsubstituted or substituted C _1-10 straight or branched chain alkyl, and unsubstituted or substituted C _1- May be substituted with one or more substituents selected from the group consisting of ₁₀ straight or branched alkoxy,

Here again, the substituted alkyl, or substituted alkoxy, may be substituted with one or more substituents selected from the group consisting of OH, halogen, cyano, nitro, amino, and C _1-3 straight or branched chain alkyl. have).

In addition, as shown in Scheme 1,

Preparing a compound represented by Chemical Formula 2 from the compound represented by Chemical Formula 3; And

It provides a method for producing a compound represented by Formula 1 of claim 1 comprising the step of preparing a compound represented by Formula 1 from the compound represented by Formula 2 prepared in the above step.

Scheme 1

(In the above Reaction Scheme 1,

, W, X, 및 Y는 제1항의 화학식 1에서 정의한 바와 같다).

, W, X, and Y are as defined in formula (1) of claim 1).

Furthermore, the present invention provides a pharmaceutical composition for the prevention or treatment of oral diseases containing the compound represented by the formula (1), stereoisomers thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In another aspect, the present invention provides a toothpaste composition for the prevention or improvement of oral diseases containing a compound represented by the formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

Furthermore, the present invention provides a goggle composition for the prevention or improvement of oral diseases containing the compound represented by the formula (1), stereoisomers thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In another aspect, the present invention provides a health functional food composition for the prevention or improvement of oral diseases containing a compound represented by the formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

Furthermore, the present invention provides a pharmaceutical composition for preventing or treating an inflammatory disease containing a compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention also provides an antibiotic composition containing the compound represented by Formula 1, its stereoisomer, or a pharmaceutically acceptable salt thereof as an active ingredient.

The novel brominated furanone derivatives or pharmaceutically acceptable salts thereof according to the present invention exhibit bacterium's quorum-sensing inhibitory activity, and can also effectively inhibit the biofilm formation of bacteria and inhibit bacterial growth. It can be used as a pharmaceutical composition, nutraceutical composition, antibiotic composition, toothpaste composition, or gargle composition containing it as an active ingredient, and is useful for oral disease or inflammatory disease such as periodontal disease such as gingivitis and periodontitis. Can be represented.

Figure 1 (a) is the OD 590nm value for each group of Example 1-13, R (furanone), positive group (Fn AI-2), negative group (Fn) in 2 μM final concentration treatment for F. nucleatum biofilm 1 (b) is a graph showing the OD 590nm value for each group in terms of percent biofilm formation (%).
(A), (b), (c) of FIG. 2 show examples 1, 11, and positive group (Fn AI-2) for F. nucleatum biofilms so that their final concentrations are 0.2 μM, 0.02 μM, and 0.002 μM. , Negative group (Fn) OD 590nm value for each group (test 1-3, a total of three repeated experiments), Figure 2 (d) shows the inhibitory effect of each group on the growth of F. nucleatum OD 600nm It is a graph confirmed by a value.
(A), (b), (c) of FIG. 3 show examples 1, 11, and positive group (Fn AI-2) for P. gingivalis biofilms so that their final concentrations are 0.2 μM, 0.02 μM, and 0.002 μM. , Negative group (Fn) OD 590nm value for each group (test 1-3, a total of three repeated experiments), Figure 3 (d) shows the inhibitory effect of each group on the growth of P. gingivalis OD 600nm It is a graph confirmed by a value.
Figure 4 (a) is a graph of the P. gingivalis biofilm formation according to the treatment of Example 1, 11 or Comparative Example 1 for the P. gingivalis biofilm so that the final concentration is 0.02 μM in terms of the control reference percentage (%) (B) is the conversion of P. gingivalis biofilm formation according to Example 1, 11 or Comparative Example 1 treatment for P. gingivalis biofilm in terms of the control percentage (%) so that the final concentration is 0.2 μM. It is shown as a graph.
(A), (b), (c) and (d) of FIG. 5 show the final concentrations of mononuclear cells (THP-1) so that the final concentrations are 2 μM, 0.2 μM, 0.02 μM, and 0.002 μM, respectively. Example 1, 11 is a bar graph showing the cell survival rate (%) of the compound, R (furanone), and ethanol treatment control (Control), Figure 5 (e) is a line graph for comparison of each group .
Figure 6 shows the inhibitory activity for each CYP isoenzyme of Example 1 compound by IC ₅₀ value.
Figure 7 shows the inhibitory activity for each CYP isoenzyme of Example 11 compound by IC ₅₀ value.
8 shows the inhibitory activity as a percentage of each CYP isoenzyme of the compound of Comparative Example 1 measured by the CYP450 screening assay.

Hereinafter, the present invention will be described in detail.

The following description is presented to aid in understanding the invention, and the present invention is not limited to the contents of the following description.

The present invention provides a compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[Formula 1]

(In the above formula 1,

X is O, S or NR ¹ ,

Wherein R ¹ is unsubstituted or substituted C _1-10 straight or branched chain alkyl, unsubstituted or substituted C _1-10 straight or branched chain alkoxy, unsubstituted or substituted C _3-7 cycloalkyl , 3 to 7 cyclic unsubstituted or substituted heterocycloalkyl, unsubstituted or substituted C _6-10 aryl containing one or more hetero atoms selected from the group consisting of N, O and S, or N, O And 5 to 10 each ring heteroaryl including one or more hetero atoms selected from the group consisting of S,

Wherein the substituted alkyl, substituted alkoxy, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl or substituted heteroaryl are hydroxy, halogen, -CN, -NO ₂ , C _1-5 May be substituted with straight or branched chain alkyl, C _1-5 straight or branched chain alkoxy;

는 비치환 또는 치환된 C_3-7의 사이클로알킬, 하나 이상의 이중결합을 포함하는 비치환 또는 치환된 C_3-7의 사이클로알케닐, N, O 및 S로 이루어진 군으로부터 선택되는 1종 이상의 헤테로 원자를 포함하는 3 내지 7각환의 비치환 또는 치환된 헤테로사이클로알킬, 비치환 또는 치환된 C_6-10의 아릴, 또는 N, O 및 S로 이루어진 군으로부터 선택되는 1종 이상의 헤테로 원자를 포함하는 5 내지 10각환의 헤테로아릴이되,

Is unsubstituted or substituted C _3-7 cycloalkyl, at least one double bond alkenyl cycloalkyl unsubstituted or substituted C _3-7 Al, including, N, O, and one or more heteroatoms selected from the group consisting of S 3 to 7-membered unsubstituted or substituted heterocycloalkyl containing atoms, unsubstituted or substituted C _6-10 aryl, or at least one hetero atom selected from the group consisting of N, O and S 5 to 10 each ring heteroaryl,

Wherein the substituted cycloalkyl, substituted cycloalkenyl, substituted heterocycloalkyl, substituted aryl, or substituted heteroaryl is a straight chain of hydroxy, halogen, -CN, -NO ₂ , C _1-10 Or branched alkyl, C _1-10 straight or branched alkoxy; And

One of W and Y is Br and the other comprises H, unsubstituted or substituted C _6-10 aryl, or a heteroaryl of one member selected from the group consisting of N, O, and S Ring or substituted 5-10 atoms of heteroaryl,

Wherein said substituted aryl, or substituted heteroaryl, is OH, halogen, cyano, nitro, amino, unsubstituted or substituted C _1-10 straight or branched chain alkyl, and unsubstituted or substituted C _1- May be substituted with one or more substituents selected from the group consisting of ₁₀ straight or branched alkoxy,

Here again, the substituted alkyl, or substituted alkoxy, may be substituted with one or more substituents selected from the group consisting of OH, halogen, cyano, nitro, amino, and C _1-3 straight or branched chain alkyl. have).

More preferably,

는 비치환 또는 치환된 C_5-6의 사이클로알킬, 하나의 이중결합을 포함하는 비치환 또는 치환된 C_5-6의 사이클로알케닐, 비치환 또는 치환된 페닐이되,

Is unsubstituted or substituted C _5-6 cycloalkyl, unsubstituted or substituted C _5-6 cycloalkenyl, unsubstituted or substituted phenyl containing one double bond,

Here, the substituted cycloalkyl, substituted cycloalkenyl, or substituted phenyl, at least one selected from the group consisting of straight chain or branched chain alkoxy of alkyl and C _1-5 straight or branched-chain C _1-5 It may be substituted with a substituent.

More preferably,

X is O; And

는

,

,

,

,

,

,

,

,

,

,

,

또는

이다.remind

Is

,

,

,

,

,

,

,

,

,

,

,

or

to be.

Further preferably,

The compound represented by Chemical Formula 1 may be a compound represented by Chemical Formula 1 ′.

[Formula 1 ']

(In Formula 1 ', each W, X, and Y are as defined in Formula 1).

Preferred examples of the compound represented by Formula 1 according to the present invention include the following compounds.

(1) (Z) -3- (bromo (phenyl) methylene) isobenzofuran-1 (3H) -one;

(2) (Z) -3- (bromo (thiophen-2-yl) methylene) isobenzofuran-1 (3H) -one;

(3) (Z) -3- (bromo (furan-2-yl) methylene) isobenzofuran-1 (3H) -one;

(4) (Z) -3- (bromo (4-chlorophenyl) methylene) isobenzofuran-1 (3H) -one;

(5) (Z) -3- (bromo (4-fluorophenyl) methylene) isobenzofuran-1 (3H) -one;

(6) (Z) -3- (bromo (4- (trifluoromethyl) phenyl) methylene) isobenzofuran-1 (3H) -one;

(7) (Z) -3- (bromo (pyridin-2-yl) methylene) isobenzofuran-1 (3H) -one;

(8) (E) -3- (bromo (pyridin-2-yl) methylene) isobenzofuran-1 (3H) -one;

(9) (Z) -3- (bromo (pyridin-3-yl) methylene) isobenzofuran-1 (3H) -one;

(10) (E) -3- (bromo (pyridin-3-yl) methylene) isobenzofuran-1 (3H) -one;

(11) (Z) -3- (bromo (pyrimidin-5-yl) methylene) isobenzofuran-1 (3H) -one; And

(12) (E) -3- (bromo (pyrimidin-5-yl) methylene) isobenzofuran-1 (3H) -one.

The compound represented by Formula 1 of the present invention may be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid, aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanes. Non-toxic organic acids such as dioate, aromatic acids, aliphatic and aromatic sulfonic acids and the like, and organic acids such as acetic acid, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid and the like. Examples of such pharmaceutically nontoxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, eye Odide, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suve Latex, sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitro benzoate, hydroxybenzoate, Methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chloro Zensulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, glycolate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene- 1-sulfonate, naphthalene-2-sulfonate, mandelate and the like.

The acid addition salt according to the present invention can be prepared by a conventional method, for example, a precipitate produced by dissolving a derivative of Formula 1 in an organic solvent such as methanol, ethanol, acetone, dichloromethane, acetonitrile and adding an organic or inorganic acid. The solvent may be prepared by filtration and drying, or the solvent and excess acid may be distilled under reduced pressure, dried, and then crystallized under an organic solvent.

Bases can also be used to make pharmaceutically acceptable metal salts. Alkali metal or alkaline earth metal salts are obtained, for example, by dissolving a compound in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is pharmaceutically suitable to prepare sodium, potassium or calcium salt as the metal salt. Corresponding salts are also obtained by reacting alkali or alkaline earth metal salts with a suitable negative salt (eg silver nitrate).

Furthermore, the present invention includes not only the compound represented by Formula 1 and pharmaceutically acceptable salts thereof, but also solvates, optical isomers, hydrates, and the like that can be prepared therefrom.

In addition, the present invention as shown in Scheme 1,

Preparing a compound represented by Chemical Formula 2 from the compound represented by Chemical Formula 3; And

It provides a method for producing a compound represented by Formula 1 of claim 1 comprising the step of preparing a compound represented by Formula 1 from the compound represented by Formula 2 prepared in the above step.

Scheme 1

(In the above Reaction Scheme 1,

, W, X, 및 Y는 제1항의 화학식 1에서 정의한 바와 같다).

, W, X, and Y are as defined in formula (1) of claim 1).

Meanwhile, the manufacturing method represented by Scheme 1 may be a manufacturing method represented by the following Scheme 1 '.

Scheme 1 '

(In the above Reaction Scheme 1 ', R is according to the definition of W, or Y in the formula (1), it can be understood as a definition other than Br).

Hereinafter, a method for preparing a compound represented by Chemical Formula 1 according to the present invention represented by Scheme 1 will be described in detail step by step.

In the method for preparing a compound represented by Formula 1 according to the present invention represented by Scheme 1, in the step of preparing a compound represented by Formula 2 from the compound represented by Formula 3,

The step is a bromination reaction, but is not limited thereto, the reaction can be carried out using CBr ₄ , the solvent usable in the step is H ₂ O, ethanol, tetrahydrofuran (THF), dichloromethane, toluene, aceto Nitrile, dimethylformamide and the like can be used, and preferably tetrahydrofuran (THF) can be used. In addition, the reaction time is not particularly limited, but preferably 10 minutes to 10 hours, 20 minutes to 2 hours, or about 30 minutes. Furthermore, the reaction temperature is not particularly limited, but is preferably performed at room temperature or 20 to 30 ° C.

In a most preferred embodiment, it can be carried out as in the following Examples of the present invention, from which the modification of the aspect, conditions and the like of the experiment and the method that can be prepared from the formula (3) of the compound represented by the formula (1) of the present invention It is included in a range.

In addition, in the preparation method represented by Scheme 1, the step of preparing a compound represented by the formula (1) from the compound represented by the formula (2), can be understood as the introduction step of W, or Y of the formula (1).

In this case, the step may be performed as in Step 2 of Example 1-12 below, it may be a manufacturing step of changing / modifying this.

In addition, the solvent that can be used in this step is not particularly limited as long as the reaction can proceed, for example, H ₂ O, ethanol, tetrahydrofuran (THF), dichloromethane, toluene, acetonitrile, dimethylform Amides and the like can be used, and preferably tetrahydrofuran (THF) can be used. In addition, the reaction time is not particularly limited, but is preferably reacted for 3-48 hours, 12-30 hours, or about 24 hours. Further, the reaction temperature is preferably, but not limited to, performed at 30-80 ° C, 50-70 ° C, or about 66 ° C.

In another aspect, the present invention provides a pharmaceutical composition for the prevention or treatment of oral diseases containing a compound represented by the formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

Here, the compound represented by Chemical Formula 1, its stereoisomer, or a pharmaceutically acceptable salt thereof has a mechanism for inhibiting quorum sensing of bacteria and a mechanism for inhibiting biofilm formation of bacteria. From this, the compound represented by Formula 1, its stereoisomers, or pharmaceutically acceptable salts thereof may inhibit the pathological activity of bacteria exposed to the site of application. Preferably in the oral cavity of the human body or mammal, it can be used as an active ingredient of a pharmaceutical composition for the prevention or treatment of oral diseases, for example, periodontal disease, specifically periodontitis, gingivitis. In addition, it can be applied to diseases such as inflammation caused by bacteria in the human body or the body of a mammal to which the above-described effects can be applied, and show useful effects, which are included in the scope of the present invention.

More closely, quorum sensing can be expressed by perceptual density recognition, where individual bacterial organisms accumulate small molecules of signaling molecules, such as autoinducers or pheromones, outside the cell to promote active proliferation. It refers to a series of biological phenomena that induce and fill quorum. Specifically, N-acyl homoserine lactone synthase protein (N-acyl homoserine lactone synthase protein) among the protein constituting the bacteria synthesizes a signaling material called normal acyl homoserine Lactone (AHL). The synthesized normal acyl homoserine lactone freely diffuses through the cell membrane during bacterial growth and accumulates in the environment outside the cell. When the density of bacteria increases and the signaling material accumulated outside the cell reaches a certain concentration, the signaling material enters the cell again and binds to a transcriptional regulator to promote expression of a specific gene. Bacteria control various physiological phenomena such as formation of biofilm, virulence, bioluminescence, production of antibiotics or delivery of tumor-induced plasmid by conjugation through the quorum sensing mechanism described above. do.

The compound represented by Formula 1 is structurally similar to N-Acyl Homoserine Lactone (AHL), which is a signaling material synthesized by bacteria, so that when it is acted around, it interferes with communication between bacteria and expresses genes. It can prevent bacterial growth as well as inhibit the formation of biofilms, which are known to increase resistance to antibiotics.

In this regard, the results of experiments were performed to evaluate the inhibitory activity of AI-2 (Autoinducer-2), which is a quorum sensing signal molecule of the compound represented by the formula (1), the stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to the present invention. In addition, the exemplary compound according to the present invention suppresses AI-2 (Autoinducer-2), which is a quorum sensing signal molecule, and has a low bioluminescenece value.

In addition, as a result of performing an experiment to evaluate the ability of the compound represented by the formula (1), the stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to the present invention to inhibit the biofilm formation of bacteria, according to the present invention, Example compounds have been shown to lower the degree of biofilm formation when incubated with bacteria.

Furthermore, the result of performing an experiment for evaluating cytotoxicity in a host cell, that is, a normal cell, of a compound represented by the formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to the present invention, all of the compounds according to the present invention There is little host cell toxicity and it has been found to be suitable for use as a drug.

In addition, an experiment for evaluating whether a compound represented by the formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to the present invention is suitable as a disease treatment drug for treating oral diseases including periodontal disease such as gingivitis periodontitis. As a result, the furanone used as a control induces an inflammatory response in the host cell, i.e., the normal cell, whereas the compound of the present invention does not induce an inflammatory response in the normal cell. Could.

On the other hand, the compound represented by the formula (1) of the present invention as a result of evaluating the inhibitory activity against the drug lyase group, it was confirmed that there is no inhibitory activity in the drug lyase group. In particular, compared to the conventional Comparative Example 1 compound has a inhibitory activity in the group of drug degrading enzymes, and problems such as drug interaction is pointed out, the compound represented by the formula (1) of the present invention is a safer compound against the problem caused by drug interaction Could be thought of as.

Thus, the novel furanone derivatives or pharmaceutically acceptable salts thereof according to the present invention exhibit excellent performance as quirumsensing antagonists that interfere with the communication between bacteria, leading to the formation of biofilms known to increase antibiotic resistance. Since it can effectively block and inhibit the growth of bacteria, it can be usefully used as a pharmaceutical composition for the prevention or treatment of oral diseases.

The compound represented by Formula 1 according to the present invention may be administered in various oral and parenteral formulations during clinical administration, and when formulated, the commonly used fillers, extenders, binders, humectants, disintegrating agents, surfactants, and the like. Prepared using diluents or excipients.

Solid form preparations for oral administration include tablets, patients, powders, granules, capsules, troches, and the like, which form at least one excipient such as starch, calcium carbonate, water, or the like. It is prepared by mixing cross, lactose or gelatin. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Liquid preparations for oral administration include suspensions, solvents, emulsions or syrups, and include various excipients such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. Can be.

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories, and the like.

As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerol, gelatin and the like can be used.

In addition, the effective dosage of the compound of the present invention to the human body may vary depending on the age, weight, sex, dosage form, health condition and degree of disease of the patient, and is generally about 0.001-100 mg / kg / day, preferably Preferably 0.01-35 mg / kg / day. Based on an adult patient weighing 70 kg, it is generally 0.07-7000 mg / day, preferably 0.7-2500 mg / day, once a day at regular intervals according to the judgment of the doctor or pharmacist. Multiple doses may be administered.

Furthermore, the present invention provides a dentifrice composition for the prevention or improvement of oral diseases containing the compound represented by the formula (1), stereoisomers thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

It provides a gargle composition for the prevention or improvement of oral diseases containing a compound represented by the formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

At this time, the toothpaste or goggle composition is a compound represented by the formula (1) of the present invention, or a pharmaceutically acceptable salt thereof is included as an active ingredient, main ingredient or auxiliary component of the quasi-drug, inhibiting quorum sensing of the above-mentioned bacteria, Biofilm formation inhibition and antimicrobial effect.

Therefore, if the compound represented by the formula (1) of the present invention, or a pharmaceutically acceptable salt thereof in addition to the toothpaste or gargle composition can be used as a quasi-drug, it is included in the scope of the present invention.

The compound represented by the formula 1 of the present invention described above has been experimentally confirmed to inhibit the quorum sensing, biofilm formation, and antifungal effect of bacteria, and is also evaluated to be safe for normal cells and safe for lytic enzymes. , Useful, and especially effective in bacteria associated with oral disease and periodontal disease.

Oral disease in the present specification, but is not limited to this, can be understood to be a disease causing lesions, such as inflammation in the oral cavity, for example, refers to various diseases occurring in the oral cavity area, the oral cavity area is The space in the mouth that connects the pharynx from the anterior lip to the posterior bulbus. In the present invention, the oral disease is a concept that includes all diseases regardless of the condition if the disease occurs in the oral cavity, non-limiting examples of oral diseases include dental caries, periodontal disease (periodonitis or gingivitis), toothache, ache, bad breath Etc. can be mentioned.

In the present invention, "dental cavities" is an infectious disease caused by bacteria inhabiting the tooth surface, also called tooth decay, and shows signs of deterioration due to erosion of the hard tissue of the tooth. Specifically, the milky, white, translucent and hard substance covering the surface of the head of the tooth and protecting the dentin of teeth is called tooth enamel or enamel. It is caused by acid caused by the decomposition of sugar and starch by the oral pathogens in the mouth. Tooth decay of the tooth is damaged and tooth decay occurs. The main cause of dental caries is dental plaque (plaque), which is a bacterial membrane formed on the surface of teeth. Saliva forms a thin, sticky membrane on the teeth, and streptococcus sobrinus, a type of streptococci, on it. Oral microorganisms, such as Streptococcus sobrinus and Streptococcus mutans, attach to form a biofilm, resulting in dendritic plaque and thickening by Fusobacterium Lose. The dental caries of the present invention includes all irrespective of the species causing the dental caries, specifically, Streptococcus mutans (Streptococcus mutans), Streptococcus sanguinis, Streptococcus sobrinus (Streptococcus sobrinus), Streptococcus ratti, Streptococcus criceti, Streptococcus anginosus and one or more selected from the group consisting of lactic acid bacteria, more specifically Streptococcus It can be a mutans.

Streptococcus mutans is a type of streptococcus gram-positive bacteria, also called cavities. The Streptococcus mutans proliferate only on the surface of the tooth, but the induction is not completed until about 30 months of age. Therefore, until this time, mutans bacteria are difficult to grow, as in infancy. If other oral bacteria settle in the mouth because they are not infected with mutans from the adult's mouth during infancy, it is difficult for new mutans to enter the already well-balanced oral ecosystem. Will be lowered. If you are already infected with the causative organism through the mouth of the adult, often brush your teeth and keep your mouth clean to help prevent tooth decay.

In the present invention, "periodontal disease" is a disease in which the gingival, periodontal ligament and bone tissue supporting the teeth are inflamed, which is also commonly referred to as vertigo, gingivitis and periodontitis depending on the severity of the disease. Divided into It is a relatively light and fast-recovering form of periodontal disease that is limited to the gums, ie soft tissues, is called gingivitis, and when this inflammation progresses to the gums and around the gum bone, it is called periodontitis. There is a V-shaped gap between the gingiva and the teeth, which attack the area below the gum line of the sulcus, releasing Lipopolysaccharide (LPS), an inflammatory stimulus. Inflammation is created, such as swelling of the gums and bleeding, which damages the periodontal ligament and adjacent tissues. As periodontitis progresses, the periodontal ligament and even the alveolar bone are damaged, resulting in loss of teeth. Malnutrition of proteins and vitamins, hormonal disorders such as pregnancy or diabetes, smoking and AIDS can make the disease worse. In addition, plaque and calculus are other causes of periodontal disease. The periodontal disease of the present invention includes all irrespective of the species causing the periodontal disease, specifically, Actinobacillus actinomycetem comitans, Porphyromonas gingivalis (Torphyromonas gingivalis), Thane At least one homogeneous member selected from the group consisting of Tannerella forsythia, Treponema denticola, and Fusobacterium nucleatum, and more specifically, porphyromonas It may be a ballis.

The Pyromonas jinjivalis is a bacteroid (Bacteroide) flexible bacteria, gram-negative bacteria, anaerobic bacteria. The porphyromonas gingivalis is found in the oral cavity in which periodontal disease has occurred, as well as in the upper gastrointestinal tract, respiratory tract and colon. In patients with chronic periodontal disease, collagen is degraded by the bacteria's collagenase enzyme, which can invade gingival fibroblasts and survive under significant concentrations of antibiotics. In addition, the fungus can invade gingival epithelial cells to survive for a long time.

In the present invention, "toothache" refers to pain in teeth when eating sweet foods or very cold or hot foods. Generally, not only the pain of the teeth itself, but also the periodontal tissue supporting the teeth in the jawbone Pain is also included. Pain usually occurs when chewing, swelling gums, and secretive smell. Toothache is a slightly different pain depending on the causative disease.In particular, the pain caused by dental caries is initially painless, but gradually progresses to deep nerve decay to the nerves in the tooth. Pain can be triggered, and if a tooth breaks or cracks, it can cause pain by stimulating the nerves as the teeth crack when touching or biting cold food. In case of pulpitis, pain is initially felt when cold food is touched, and when it is further developed, pain is felt in hot food, and when inflammation progresses and pulp tissue dies, there is no reaction to cold or hot root tooth (tip of tooth root) Pain caused by inflammation.

In the present invention, "silin" refers to a hypersensitive dentine, and a symptom refers to dentin hyperesthesia. Sirin's symptoms are sensitive when the dentin of exposed teeth is in contact with cold air or irritating foods and is present in 60-98% of adults with periodontal disease. This symptom is caused by a tooth decay on the gum side due to incorrect brushing habits, excessive occlusal force, or acid dissolution, poor oral hygiene conditions, periodontal treatment, restorative treatment, and acidification of the tooth. May appear. As a fundamental cause of the symptoms, many of the tubules present in the dentin of the tooth are exposed to the outside. By exposing all the stimuli to the nerves in the pulp, they are more sensitive to the same stimulus and can cause pain. have. Syringe symptoms can range from mild to intense, painful pain, and because of the nature of the teeth do not regenerate, painkillers and anti-inflammatory drugs are not a fundamental solution to the symptoms. Shirini symptoms can be seen throughout the entire tooth and may be confined to a specific area, such as the maxilla or mandible, or right or left. The most common areas are different depending on the cause, but mainly the fangs and small molar areas, the most severe pain area is more than 90% of the cervical area, the boundary between the gum and teeth.

In the present invention, " bad breath " is an odor derived from the oral cavity and adjacent organs, and 85-90% of the bad breath is derived from the oral cavity, particularly from the back of the tongue. The main components of bad breath are volatile sulfur compounds, of which 90% of the total volatile sulfur compounds are hydrogen sulfide made from cysteine and methyl mercaptan and dimethyl sulfide made from methionine. These components are mainly produced by protein enzymes secreted by anaerobic bacteria, and the back of the tongue is the most important habitat. This area is not well cleaned by saliva, and there are many small depressions, which is a place where bacteria live continuously. The generation of volatile sulfur compounds by anaerobic bacteria is the most important cause of bad breath, but is also caused by oral diseases such as dental caries, periodontitis and dry mouth. Many types of anaerobic bacteria are involved in the development of bad breath, and non-limiting examples of the cause of bad breath include Porphyromonas gingivalis, which secrete many types and large amounts of enzymes.

The quasi-drug composition of the present invention may include an oral quasi-drug. Ingredients included in the quasi-drug composition of the present invention may include components commonly used in the quasi-drug composition for oral cavity as an active ingredient, for example, abrasives, wetting agents, binders, foaming agents, sweeteners, preservatives, medicinal ingredients, flavors Agents, dyes, solvents, brighteners, solubilizers or pH adjusters.

Oral quasi-drug composition of the present invention can be prepared in any formulation commonly prepared in the art, for example, toothpaste, mouthwash, mouthwash, gum, candy, oral spray, oral ointment, oral varnish It may have a formulation such as mouthwash, gum massage cream, but is not limited thereto.

As an example, when the oral quasi-drug composition of the present invention is a toothpaste formulation, it may include a humectant, an abrasive, a binder, a foaming agent, a flavoring agent, a sweetening agent, a coloring agent, a preservative, an active ingredient, a solvent, a pH adjusting agent, and the like.

The humectant is to make the powder in the toothpaste component into paste form and to prevent the toothpaste from hardening in the air. Glycerin, sorbitan, propylene glycol, polyethylene glycol, etc., alone or in combination of two or more kinds of the total weight of the composition 1-60 %, Specifically 10-50% by weight can be used.

The foaming agent enhances the cleaning effect by dispersing the toothpaste in the oral cavity, and acts as a surfactant to clean the oral contamination. Surfactants such as sodium lauryl sulfate, sodium lauryl sodium succinate, sodium alkyl sulfovacate, and sucrose fatty acid ester are used alone. Alternatively, two or more thereof may be mixed to use 0.5-10% by weight, specifically 0.5-5% by weight of the total weight of the composition.

The binder is to prevent separation between the powder and the liquid component in the toothpaste, cellulose derivatives such as carboxymethyl cellulose sodium, methyl cellulose, hydroxy propyl cellulose and sodium alginate, carrageenan, xanthan gum, etc. alone or in combination of two or more 0.1-5% by weight, specifically 0.3-2% by weight, of the total weight of the composition can be used.

The abrasive removes the attachment of the tooth surface without injuring the tooth surface and makes the original polish. The calcium carbonate (CaCO3), dicalcium phosphate (CaHPO4, CaHPO42H2O), silicic anhydride (SiO22H2O), aluminum hydroxide (Al) (OH) 3), potassium pyrophosphate, magnesium carbonate, or the like may be used alone or in combination of two or more, 1-60% by weight, specifically 10-50% by weight, of the total weight of the composition.

The flavoring agent is to enhance the feeling of use by providing a refreshing and odor to the toothpaste, peppermint oil, spearmint oil, menthol, etc. alone or in combination of two or more kinds of 1-60% by weight, specifically 0.01- 5% by weight can be used.

The sweetener is to remove the unpleasant taste caused by the raw material of the toothpaste, and to improve the refreshing feeling. It is 1-60% by weight of the total weight of the composition by mixing alone or two or more of saccharic acid, aspartame, xylitol, and licorice. 0.01-5% by weight can be used.

On the other hand, a health functional food composition for the prevention or improvement of oral diseases containing a compound represented by the formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient may be provided.

In addition to the active ingredient included in the health functional food composition of the present invention may include a food supplement acceptable food supplement.

In the present invention, the "food supplement" means a component that can be added to food supplements, and can be appropriately selected and used by those skilled in the art as being added to prepare the health functional food of each formulation. Examples of food additives include flavors such as various nutrients, vitamins, minerals (electrolytes), synthetic and natural flavors, colorants and fillers, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners. , pH regulators, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like, but is not limited to the kind of food additives of the present invention by the above examples.

The "health functional food" of the present invention refers to a food prepared and processed in the form of tablets, capsules, powders, granules, liquids and pills using raw materials or ingredients having useful functions for the human body. Here, the term "functionality" means obtaining useful effects on health purposes such as nutrient control or physiological effects on the structure and function of the human body. The health functional food of the present invention can be prepared by a method commonly used in the art, and the preparation can be prepared by adding raw materials and ingredients commonly added in the art. In addition, the formulation of the health functional food can also be prepared without limitation as long as the formulation is recognized as a health functional food.

Furthermore, the present invention provides a pharmaceutical composition for preventing or treating an inflammatory disease containing a compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In this case, the inflammatory disease may be understood as an inflammatory response itself in vivo, or a disease induced or induced therefrom, or as a disease indicating inflammation in a medical sense as a symptom.

In one aspect of the invention,

The inflammatory disease may be, for example, inflammation of mucous and mucocutaneousus tissues of the oral cavity and dental pulp, or oral bacterial infection.

In one embodiment of the invention,

The bacteriostatic inflammation may be understood to be inflammation caused by bacteria, in particular, confirmed by experiments in the following Examples or Experimental Examples. Compound represented by the formula (1) of the present invention, its stereoisomers or pharmaceutically acceptable salts thereof can excellently inhibit the quorum sensing, biofilm formation, bacterial growth of the bacteria, even in the above inflammatory diseases Can be effective.

The present invention also provides an antibiotic composition containing the compound represented by Formula 1, its stereoisomer, or a pharmaceutically acceptable salt thereof as an active ingredient.

In one aspect of the invention,

The antibiotic composition is an antibiotic commonly understood in the medical field, and may be understood as, for example, a component or agent for inhibiting bacteria, bacteria, microorganisms, and the like, and the antibiotic composition may be, for example, an antibacterial or antifungal agent. Can be understood.

In one embodiment of the invention,

The antibiotic composition is a compound represented by the formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, which is an active ingredient of the present invention, in particular, quorum sensing for bacteria as confirmed in the following Examples or Experimental Examples, It can be understood that it can be provided as an antibiotic composition, which can suppress the biofilm formation and bacterial growth excellently.

Hereinafter, the present invention will be described in detail by Examples and Experimental Examples.

However, the following Examples and Experimental Examples are merely illustrative of the present invention, but the content of the present invention is not limited thereto.

Example 1 Preparation of (Z) -3- (Bromo (phenyl) methylene) isobenzofuran-1 (3H) -one

Step 1: Preparation of 3- (dibromomethylene) isobenzofuran-1 (3H) -one

Triisopropylphosphite (2.7 ml, 5.00 mmol) in 10 mL of THF containing Zn dust (490 mg, 7.50 mmol) was cooled to 0 ° C. under Ar. A solution of CBr ₄ (2.49 g, 7.50 mmol) in THF was added slowly to a solution of P (i-PrO) ₃ and the reaction was stirred until the color turned yellow. Then a solution of phthalic anhydride in THF was added. The reaction was stirred at rt for 30 min. sat. After quenching by addition of aq NaHCO ₃ solution (2.5 mL), the phases were then separated and the crude product was extracted from the aqueous layer with ether. The residue was dissolved in ether and filtered through a pad of celite. The resulting filtrate was concentrated by rotary evaporation and purified by column chromatography (1-5% ether in hexanes) to afford the desired compound.

Yield 62%. ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 8.36 (d, J = 8.0 Hz, 1H), 7.97 (d, J = 7.7 Hz, 1H), 7.91 (t, J = 8.0 Hz, 1H), 7.75 (t, J = 8.0 Hz, 1H). ¹³ C NMR (DMSO-d ₆ , 100 MHz) δ 164.53, 146.01, 136.32, 136.17, 131.93, 126.55, 125.86, 124.24, 77.69.

Step 2: Preparation of (Z) -3- (Bromo (phenyl) methylene) isobenzofuran-1 (3H) -one

Under argon atmosphere, the compound prepared in step 1 (152 mg, 0.5 mmol), AsPh ₃ (150 mg, 0.1 mmol), Pd (dba) ₂ (14 mg, 0.025 mmol), and THF (4 ml) were ovened -Added to dry Schlenk tube. Phenyl-SnBu ₃ (tributyl (phenyl) stanan) (0.25 mmol) was added to the stirred mixture, then the mixture was heated at 66 ° C. for 24 hours. The reaction mixture was cooled to rt and extracted with DCM (10 mL), washed with brine, dried over MgSO ₄ and filtered. The crude product was purified by flash chromatography on silica gel to afford the desired compound.

Yield 72%. ¹ H NMR 7 (DMSO-d ₆ , 500 MHz) δ 7.94 (d, J = 8.0 Hz, 1H), 7.66-7.54 (m, 7H,), 6.56 (d, J = 7.8 Hz, 1H). ¹³ C NMR (DMSO-d ₆ , 100 MHz) δ 165.56, 144.65, 137.05, 136.15, 135.62, 131.27, 130.87, 130.15, 129.97, 126.11, 125.32, 122.47, 104.65. for [M + Na] + C ₁₅ H ₉ BrNaO ₂ ⁺ : 322.9678; found 322.9678.

Example 2 Preparation of (Z) -3- (Bromo (thiophen-2-yl) methylene) isobenzofuran-1 (3H) -one

Step 1: Preparation of 3- (dibromomethylene) isobenzofuran-1 (3H) -one

In the same manner as in Step 1 of Example 1, the target compound was prepared.

Step 2: Preparation of (Z) -3- (Bromo (thiophen-2-yl) methylene) isobenzofuran-1 (3H) -one

Performed as in Step 2 of Example 1 above, in place of phenyl-SnBu ₃ (tributyl (phenyl) stanan), thiophen-2-yl-SnBu ₃ (tributyl (thiophen-2-yl) star I) was used to prepare the target compound.

Yield 65%. ¹ H NMR (DMSO-d ₆ , 500 MHz) δ 7.97 (d, J = 5.2 Hz, 2H), 7.72-7.64 (m, 2H), 7.50-7.44 (m, 1H), 7.26 (t, J = 4.3 Hz, 1H), 6.84 (d, J = 7.0 Hz, 1H). ¹³ C NMR (DMSO-d ₆ , 100 MHz) δ 165.25, 146.30, 137.14, 136.74, 135.82, 131.69, 131.59, 131.40, 128.59, 126.18, 125.38, 122.66, 96.26. for [M + Na] + C ₁₃ H ₇ BrNaO ₂ S ⁺ : 328.9242; found 328.9242.

Example 3 Preparation of (Z) -3- (Bromo (furan-2-yl) methylene) isobenzofuran-1 (3H) -one

Step 1: Preparation of 3- (dibromomethylene) isobenzofuran-1 (3H) -one

In the same manner as in Step 1 of Example 1, the target compound was prepared.

Step 2: Preparation of (Z) -3- (Bromo (furan-2-yl) methylene) isobenzofuran-1 (3H) -one

Performed as in Step 2 of Example 1 above, in place of phenyl-SnBu ₃ (tributyl (phenyl) stanan), furan-2-yl-SnBu ₃ (tributyl (furan-2-yl) stanan) Using to prepare the target compound.

Yield 62%. ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 8.02 (s, 1H), 7.98 (d, J = 7.5 Hz, 1H), 7.82-6.68 (m, 2H), 7.34 (d, J = 7.9 Hz, 1H), 7.03 (d, J = 3.3 Hz, 1H), 6.80-6.74 (m, 1H). ¹³ C NMR (DMSO-d ₆ , 125 MHz) δ 165.13, 146.74, 146.04, 136.34, 135.96, 131.88, 126.24, 125.45, 123.02, 115.35, 112.89, 93.11, 79.25. for [M + Na] + C 13 H 7 BrNaO 3 +: 312.9470; found 312.9471.

Example 4 Preparation of (Z) -3- (Bromo (4-chlorophenyl) methylene) isobenzofuran-1 (3H) -one

Step 1: Preparation of 3- (dibromomethylene) isobenzofuran-1 (3H) -one

In the same manner as in Step 1 of Example 1, the target compound was prepared.

Step 2: Preparation of (Z) -3- (Bromo (4-chlorophenyl) methylene) isobenzofuran-1 (3H) -one

Perform as in Step 2 of Example 1, using 4-chlorophenyl-SnBu ₃ (tributyl (4-chlorophenyl) stanan) instead of phenyl-SnBu ₃ (tributyl (phenyl) stanan) To prepare the target compound.

Yield 59%. ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 7.99-7.93 (m, 1H), 7.71-7.59 (m, 6H), 6.83-6.50 (m, 1H). ¹³ C NMR (DMSO-d ₆ , 125 MHz) δ 165.49, 145.00, 136.88, 135.83, 135.58, 135.04, 132.16, 131.41, 130.13, 126.17, 125.34, 122.62, 103.14. for [M + H] + C ₁₅ H ₉ BrClNaO ₂ ⁺ : 334.9469; found 334.9469.

Example 5 Preparation of (Z) -3- (Bromo (4-fluorophenyl) methylene) isobenzofuran-1 (3H) -one

Step 1: Preparation of 3- (dibromomethylene) isobenzofuran-1 (3H) -one

In the same manner as in Step 1 of Example 1, the target compound was prepared.

Step 2: Preparation of (Z) -3- (Bromo (4-fluorophenyl) methylene) isobenzofuran-1 (3H) -one

Performed as in Step 2 of Example 1 above, instead of phenyl-SnBu ₃ (tributyl (phenyl) stanan), 4-fluorophenyl-SnBu ₃ (tributyl (4-fluorophenyl) stanan) Using to prepare the target compound.

Yield 75%. ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 7.96-7.90 (m, 1H), 7.68-7.58 (m, 4H), 7.46-7.38 (m, 2H), 6.64-6.56 (m, 1H). ¹³ C NMR (DMSO-d ₆ , 125 MHz) δ 165.53, 163.40 (d, J = 248.6 Hz), 144.93, 136.98, 135.77, 132.73 (d, J = 8.9 Hz), 132.56, 131.33, 126.14, 125.32, 122.58 , 117.15 (d, J = 22.0 Hz), 103.43.for [M + Na] + C 15 H 8 BrFNaO 2 +: 340.9584; found 340.9584.

Example 6 Preparation of (Z) -3- (Bromo (4- (trifluoromethyl) phenyl) methylene) isobenzofuran-1 (3H) -one

Step 1: Preparation of 3- (dibromomethylene) isobenzofuran-1 (3H) -one

In the same manner as in Step 1 of Example 1, the target compound was prepared.

Step 2: Preparation of (Z) -3- (Bromo (4- (trifluoromethyl) phenyl) methylene) isobenzofuran-1 (3H) -one

In the same manner as in Step 2 of Example 1, except for phenyl-SnBu ₃ (tributyl (phenyl) stanan), 4- (trifluoromethyl) phenyl-SnBu ₃ (tributyl (4- (trifluoro) Romethyl) phenyl) stanan) was used to prepare the target compound.

Yield 67%. ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 7.99-7.94 (m, 3H), 7.85 (s, 1H), 7.83 (s, 1H), 5 7.68-7.62 (m, 2H), 6.66-6.62 ( m, 1 H). ¹³ C NMR (DMSO-d ₆ , 125 MHz) δ 165.40, 145.36, 140.33, 136.74, 135.88, 131.56, 131.31, 131.04 (q, J = 11.25 Hz), 126.94 (q, J = 3.75 Hz), 126.24, 125.43, 124.19 (q, J = 268.75 Hz), 122.57, 102.51. for [M + H] + C 16 H 9 BrF 3 O 2 +: 368.9733; found 368.9733.

Example 7 Preparation of (Z) -3- (Bromo (pyridin-2-yl) methylene) isobenzofuran-1 (3H) -one

Step 1: Preparation of 3- (dibromomethylene) isobenzofuran-1 (3H) -one

In the same manner as in Step 1 of Example 1, the target compound was prepared.

Step 2: Preparation of (Z) -3- (Bromo (pyridin-2-yl) methylene) isobenzofuran-1 (3H) -one

Performed as in Step 2 of Example 1 above, but instead of phenyl-SnBu ₃ (tributyl (phenyl) stanan), pyridin-2-yl-SnBu ₃ (tributyl (pyridin-2-yl) stanan) Using to prepare the target compound.

Yield 48%. ¹ H NMR (DMSO-d ₆ , 500 MHz) δ 8.78 (d, J = 4.5 Hz, 1H), 8.04 (t, J = 7.7 Hz, 1H), 7.96 (d, J = 7.3 Hz, 1H), 7.78 (d, J = 7.8 Hz, 1H), 7.71-7.55 (m, 3H), 6.65 (d, J = 7.7 Hz, 1H). ¹³ C NMR (DMSO-d ₆ , 125 MHz) δ 165.28, 153.65, 150.67, 145.79, 138.54, 136.66, 131.61, 126.14, 126.04, 125.50, 125.43, 125.43, 122.94, 104.07. for [M + Na] + C 14 H 8 BrNNaO 2 +: 323.9631; found 323.9631.

Example 8 Preparation of (E) -3- (Bromo (pyridin-2-yl) methylene) isobenzofuran-1 (3H) -one

Step 1 and Step 2 of Example 7 were carried out in the same manner, but the (E) -isomer, which is not the (Z) -isomer of Example 7, was obtained as a final compound of interest.

Yield 32%. ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 8.75-8.68 (m, 2H), 8.09-7.77 (m, 5H), 7.44 (m, 1H). ¹³ C NMR (CDCl ₃ -d, 125 MHz) δ 165.56, 153.10, 149.84, 144.72, 138.51, 136.45, 135.01, 131.26, 126.10, 125.99, 125.95, 125.70, 123.71, 108.37. for [M + H] + C 14 H 9 BrNO 2 +: 301.9811; found 301.9811.

Example 9 Preparation of (Z) -3- (Bromo (pyridin-3-yl) methylene) isobenzofuran-1 (3H) -one

Step 1: Preparation of 3- (dibromomethylene) isobenzofuran-1 (3H) -one

In the same manner as in Step 1 of Example 1, the target compound was prepared.

Step 2: Preparation of (Z) -3- (Bromo (pyridin-2-yl) methylene) isobenzofuran-1 (3H) -one

Performed as in Step 2 of Example 1, but instead of phenyl-SnBu ₃ (tributyl (phenyl) stanan), pyridin-3-yl-SnBu ₃ (tributyl (pyridin-3-yl) stanan) Using to prepare the target compound.

Yield 32%. ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 8.86-8.70 (m, 2H), 8.06 (d, J = 7.9 Hz, 1H), 7.98 (d, J = 6.5 Hz, 1H), 7.73-7.52 ( m, 3H), 6.59 (d, J = 7.0 Hz, 1H). ¹³ C NMR (CDCl ₃ -d, 100 MHz) δ 165.27, 150.96, 150.26, 145.72, 137.70, 136.79, 134.69, 130.60, 126.03, 125.78, 122.19, 100.96. for [M + H] + C 14 H 9 BrNO 2 +: 301.9811; found 301.9811.

Example 10 Preparation of (E) -3- (Bromo (pyridin-3-yl) methylene) isobenzofuran-1 (3H) -one

Step 1 and Step 2 of Example 9 were carried out in the same manner, but the (E) -isomer, which is not the (Z) -isomer of Example 9, was obtained as a final compound of interest.

Yield 12.8%. ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 8.88 (s, 1H), 8.67 (d, J = 8.0 Hz, 1H), 8.61 (d, J = 3.7 Hz, 1H), 8.12-7.96 (m, 3H), 7.83 (t, J = 7.4 Hz, 1H), 7.60-7.52 (m, 1H). ¹³ C NMR (CDCl ₃ -d, 125 MHz) δ 165.31, 150.52, 149.59, 143.59, 143.86, 137.92, 137.17, 134.82, 131.05, 125.99, 125.60, 125.39, 104.09. for [M + H] + C 14 H 9 BrNO 2 +: 301.9811; found 301.9811.

Example 11 Preparation of (Z) -3- (Bromo (pyrimidin-5-yl) methylene) isobenzofuran-1 (3H) -one

Step 1: Preparation of 3- (dibromomethylene) isobenzofuran-1 (3H) -one

In the same manner as in Step 1 of Example 1, the target compound was prepared.

Step 2: Preparation of (Z) -3- (Bromo (pyridin-2-yl) methylene) isobenzofuran-1 (3H) -one

Performed as in Step 2 of Example 1 above, but instead of phenyl-SnBu ₃ (tributyl (phenyl) stanan), pyrimidin-5-yl-SnBu ₃ (tributyl (pyrimidin-5-yl) star I) was used to prepare the target compound.

Yield 45%. ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 9.40 (s, 1H), 9.12 (s, 2H), 8.06-7.94 (m, 1H), 7.76-7.60 (m, 2H), 6.74 (d, J = 6.8 Hz, 1H). ¹³ C NMR (DMSO-d ₆ , 125 MHz) δ 165.26, 159.72, 158.30, 146.62, 136.58, 136.07, 131.83, 131.38, 126.43, 125.37, 122.64, 97.16. for [M + H] + C ₁₃ H ₈ BrN ₂ O ₂ ⁺ : 302.9764; found 302.9764.

Example 12 Preparation of (E) -3- (Bromo (pyrimidin-5-yl) methylene) isobenzofuran-1 (3H) -one

Step 1 and Step 2 of Example 11 were carried out in the same manner, but the (E) -isomer of Example 11, which is not the (Z) -isomer of Example 11, was obtained as a final compound of interest.

Yield 7.6%. ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 9.21 (s, 1H), 9.13 (s, 2H), 8.68 (d, J = 8.0 Hz, 1H), 8.12-7.98 (m, 2H), 7.86 ( t, J = 7.5 Hz, 1H). ¹³ C NMR (CDCl ₃ -d, 125 MHz) δ 158.15, 158.11, 157.29, 137.56, 135.03, 131.54, 131.18, 130.93, 126.20, 125.51, 125.49, 100.05. for [M + H] + C ₁₃ H ₈ BrN ₂ O ₂ ⁺ : 302.9764; found 302.9764.

Comparative Example 1 Preparation of 3- (dibromomethylene) -7-methylisobenzofuran-1 (3H) -one

Triphenylphosphine (1.6 g, 6.0 mmol) was dissolved in 6 ml of THF and cooled to 0 ° C. under nitrogen gas. To this, a THF (5.0 ml) solution in which CBr ₄ (1.0 g, 3.0 mmol) was dissolved was added, and stirred until the color of the solution turned yellow. After discoloration, TEA (1.08 ml, 6.0 mmol) was added dropwise for 5 minutes, and then a solution of THF (1 ml) in which 3-methylphthalic anhydride (0.16 g, 1.0 mmol) was dissolved was added dropwise. After the reaction solution was stirred at 0 ° C. for 30 minutes, the reaction solution was brought to room temperature and then stirred overnight. The reaction was quenched with saturated aqueous NH ₄ Cl solution. Then, when the phases were separated, the aqueous layer was extracted with hexane. The combined organic layers were concentrated under reduced pressure. The residue obtained was taken up in ethoxyethane and filtered through a pad of celite. After the procedure described above, the resulting solution was concentrated by rotary evaporation and purified by flash column chromatography to afford the desired compound (0.097 g, 31%).

¹ H NMR (DMSO-d ₆ , 500 MHz) δ 8.18 (d, J = 7.9 Hz, 1H), 7.77 (t, J = 7.7 Hz, 1H), 7.55 (d, J = 7.5 Hz, 1H), 2.61 (s, 3 H). ¹³ C NMR (DMSO-d ₆ , 125 MHz) δ 164.0, 145.2, 139.8, 136.1, 135.2, 132.9, 122.8, 121.3, 76.5, 16.9.

The compounds prepared in Examples 1 to 12 are shown in Table 1 below.

Example constitutional formula Example constitutional formula One

7

2

8

3

9

4

10

5

11

6

12

Experimental Example 1 Evaluation of Bacterial Biofilm Formation Inhibitory Activity

In order to evaluate the inhibitory activity of bacterial biofilm formation of the compounds according to the present invention, the following experiments were carried out.

Specifically, sterilized cover glass slips (round, 12 mm radius) were placed on the cell culture plate (24 well plate) at the bottom of each well using tweezers. After adding 10% of the total volume of F. nucleatum AI-2 to the medium in which the bacteria are to be cultured (BHI medium + supplement), the final concentration of each compound was diluted by 2 μM, 0.2 μM, It was prepared to be 0.02 μM, 0.002 μM (BHI, a component of the medium is Brain Heart Infusion Medium, and hemin (10 μg / ml) and vitamin K (0.2 μg / ml) were used as a supplement).

The resulting cultures were placed in plate wells with glass slip and bacteria were inoculated to each well as desired (2 × 10 ⁷ cells / ml for F. nucleatum and 4 × 10 ⁸ cells / ml for P. gingivalis). ).

In the case of the positive control group (Fn AI-2) not treated with the compound of the present invention, F. nucleatum AI-2 was added to the bacterial culture medium (BHI + supplement), and the same number of bacteria were inoculated respectively. .

On the other hand, in the case of the negative control group (Fn) not treated with both the compound and F. nucleatum AI-2, the same amount of PBS was added to the bacterial culture medium (BHI + supplement) instead of F. nucleatum AI-2.

The completed well plates were maintained for 48 hours at 37 ° C., anaerobic conditions (10% H ₂ , 10% CO ₂ and 80% N ₂ ) and each bacteria were incubated. Biofilm formed on the glass slip by bacteria cultured in each group was quantified and compared using the following technique.

<1-1> Crystal Violet Dyeing Technique

The glass slip on which the biofilm was formed was stained with 1% crystal violet solution for 10 minutes, washed three times with PBS (Phosphate buffered saline) solution, and then decolorized with acetone-alcohol. 200 μl of the decolorizing solution containing crystal violet was added to the microplate by group, and then the OD 590nm value was measured and compared and analyzed by the microplate reader (Microplate reader, Model 550, Bio-Rad, USA). The higher the biofilm increased by the molecules secreted by nucleatum or P. gingivalis, the smaller the measured value is.

Experimental results for F. nucleatum in the above experiments are shown in Table 2 and FIGS.

FIG. 1 (a) shows OD 590 nm values of Examples 1-12, R (furanone), positive group (Fn AI-2), and negative group (Fn) at 2 μM for the F. nucleatum biofilm. FIG. 1B is a graph showing the OD 590 nm value for each group in terms of percent biofilm formation (%).

Table 2 shows the numerical value of the percentage of the F. nucleatum biofilm formation for each group.

Biofilm Formation Rate (%) Fn AI-2 100 R (furanone) 33.26418 Example 1 32.57261 Example 2 56.98479 Example 3 47.37206 Example 4 50.20747 Example 5 61.75657 Example 6 92.04703 Example 7 39.34993 Example 8 60.99585 Example 9 39.14246 Example 10 40.45643 Example 11 38.10512 Example 12 47.16459

Looking at Figure 1 and Table 1, all of the compound according to the present invention is confirmed to have excellent inhibitory activity against bacterial biofilm formation.

2 is an additional experiment to evaluate the results of Example 1 and Example 11 confirmed the excellent bacterial biofilm inhibitory activity from the results confirmed in Table 1 and Figure 1, Figure 2 (a) is the final concentration Example 1, 11, positive group (Fn AI-2), negative group (Fn) for each group of OD 590nm values for the F. nucleatum biofilm to be 0.2 μM, 0.02 μM, and 0.002 μM (total 3) Repeated experiments), Figure 2 (b) is a graph confirming the inhibitory effect of each group on the growth of F. nucleatum with an OD 600nm value.

FIG. 3 (a) shows examples 1, 11, positive group (Fn AI-2) and negative group (Fn) for each group of P. gingivalis biofilms so that final concentrations are 0.2 μM, 0.02 μM, and 0.002 μM. It is a graph showing the OD 590nm value (three repeated experiments), and (b) of FIG.

Referring to Figures 2 and 3, Examples 1 and 11 according to the present invention can be confirmed that both excellent suppression of biofilm formation of F. nucleatum and P. gingivalis even at a concentration of 0.002 μM, and also bacterial growth is preferably suppressed can confirm.

Figure 4 (a) is a graph of the P. gingivalis biofilm formation according to the treatment of Example 1, 11 or Comparative Example 1 for the P. gingivalis biofilm so that the final concentration is 0.02 μM in terms of the control reference percentage (%) (B) is the conversion of P. gingivalis biofilm formation according to Example 1, 11 or Comparative Example 1 treatment for P. gingivalis biofilm to the final reference concentration (%) so that the final concentration is 0.2 μM. It is shown as a graph.

Referring to Figure 4, Examples 1 and 11 according to the present invention is confirmed to suppress the formation of P. gingivalis biofilm better than Comparative Example 1.

Therefore, the compound according to the present invention can effectively inhibit the quorum sensing, biofilm formation, and growth of bacteria, as a pharmaceutical composition or a nutraceutical composition containing it as an active ingredient, or as a quasi-drug composition such as toothpaste and gargle. It can be seen that there is a prophylactic, ameliorating, or therapeutic effect useful for oral diseases, for example, periodontal diseases such as gingivitis, periodontitis.

<1-2> Confocal Scanning Laser Microscopy

The glass slip on which the biofilm was formed was stained using a live / dead-BacLight bacterial viability kit (Invitrogen, Grand Island, NY, USA), followed by confocal scanning laser microscopy (Olympus FV300, Tokyo, Japan). Morphology was observed and compared.

Experimental Example 2 Cytotoxicity Evaluation

In order to evaluate the toxicity of the compounds according to the present invention on normal cells, the following experiments were carried out.

Experimental Example 2-1 Cytotoxicity Evaluation in Human Monocyte Cell Line (THP-1)

In order to evaluate whether the compound of the present invention showing excellent biofilm inhibitory effect of periodontal pathogens is toxic to host cells (normal cells), human mononuclear cells (THP-1,1) in each well of a 96 well plate × 10 ⁵ cells / well), respectively, by treating the concentrations of R (furanone), Example 1 and Example 11 to a final concentration of 2 μM, 0.2 μM, 0.02 μM, or 0.002 μM, the finished plate was then treated at 37 ° C. Incubated for 24 hours. After 24 hours of incubation, 10 μl of CCK-8 (cell counting kit-8, Dojindo Molecular Technologies, Inc) solution was added to each well equally, and then incubated in a 37 ° C. incubator for 1-4 hours. Cell viability was evaluated by measuring the OD 450 nm value at 1 hour intervals during the above, and the results are shown in FIG. 4.

Figure 4 is a graph showing the cell viability (%) of Example 1, 11 compounds of the present invention, R (furanone), and ethanol treatment control (Control) for monocytes (THP-1).

Referring to Figure 4, it was shown that the example compound of the present invention has little toxicity against monocyte cells.

Therefore, the compound of the present invention can inhibit the quorum sensing and biofilm formation of bacteria excellently, and it appears that there is little toxicity to normal cells, pharmaceutical composition or health functional food composition, or toothpaste, containing it as an active ingredient, It may be usefully used as a quasi-drug composition such as gargle.

Experimental Example 3 CYP Screening Analysis

The degree of inhibition of CYP450 enzyme activity was measured to predict and evaluate the drug-drug interactions of the compounds according to the invention.

This experiment evaluated the inhibitory activity of the compound of the present invention on the drug metabolic enzyme in CYP3A4, CYP2C9, CYP1A2, CYP2E1, CYP2D6, CYP2C19, etc., which are known to be involved in drug metabolism in humans.

<Analysis of Inventive Example 1 Compounds>

CYP450 screening assay of the compound of Example 1 of the present invention was performed as follows.

a) sample processing

Human liver microsomes (0.25 mg / ml) and 0.1M phosphate buffer (pH 7.4)

Substrate Drug Cocktail (Phenacetin 50 μM, Diclofenac 10 μM, S-mephenytoin 100

μM, Dextromethorphan 5 μM, Midazolam 2.5 μM) and compound QS-LLZ-1, respectively.

NADPH generation after addition of 0,2,5,10,20 μM concentration and pre-incubation at 37 ° C. for 5 minutes.

The system solution was added and incubated at 37 ° C. for 15 minutes. To end the reaction

Add acetonitrile solution containing Terfenadine and centrifuge for 5 minutes

(14,000 rpm, 4 ℃) and the supernatant was injected into the LC-MS / MS system to metabolize the substrate drug

Simultaneous analysis evaluated drug metabolism inhibitory ability by QS-LLZ-1.

b) LC-MS / MS analysis

The metabolite of each CYP isoenzyme indicator drug produced through the reaction was Shimadzu Nexera.

Analysis using XR system and TSQ vantage (Thermo). HPLC column is Kinetex EVO

C18 column (2.1 × 100 mm, 2.6μm particle size; Phenomenex, USA) was used,

Mobile phase contains distilled water containing 0.1% formic acid (A) and acetonitrile containing 0.1% formic acid (B)

The Gradient program was used as follows.

Minutes Flow rate (mL / min) % A % B 0 0.3 100 0 1.0 0.3 60 40 4.0 0.3 50 50 4.1 0.3 100 0 7.0 0.3 100 0

The generated metabolites were quantified using MRM (Multiple Reaction Monitoring) quantification mode and data analysis was performed using Xcalibur (version 1.6.1). Experimental results are shown in Figure 6 by the IC ₅₀ value of the inhibitory activity of each CYP isoenzyme activity by the compound of Example 1 of the present invention obtained as% activity for the control without the inhibitor.

Referring to Figure 6, the compound of Example 1 shows a strong inhibitory activity of more than 50% (IC ₅₀ <2 μM) against CYP1A2, it is necessary to consider the drug interaction with the drug metabolized by the isoenzyme Confirmed. There is no significant result for other isoenzymes, so it can be used as a drug having a low effect due to drug interaction and ensuring safety.

Analysis of Compound of Inventive Example 11

Except for using the compound of Example 11 in place of the compound of Example 1, the experiment was carried out in the same manner as in Example 1, and the results of the experiment are the respective CYP isoenzyme activity by the compound of Example 11 of the present invention Inhibitory activity of is shown in FIG. 7 as IC ₅₀ value obtained as% activity for the control without the inhibitor.

Referring to Figure 7, the compound of Example 11 shows a strong inhibitory activity of more than 50% (IC ₅₀ = 3.0 μM) against CYP1A2, it is necessary to consider the drug interaction with the drug metabolized by the isoenzyme Confirmed. There is no significant result for other isoenzymes, so it can be used as a drug having a low effect due to drug interaction and ensuring safety.

<Analysis of Comparative Compounds>

The CYP450 screening assay of the compound of Comparative Example 1 is changed (metabolized) to luciferin, a luminescent substrate, when a reaction occurs by enzyme and NADPH regeneration using a luminescent CYP substrate, which is a P450-Glo ™ substrate. The luciferin detection reagent is then added to measure luminescence, thereby measuring the activity of the CYP enzyme.

As an experimental material,

(1) Positive Control Inhibitor (Stock solution in DMSO)

CYP1A2: 10 mM α-naphthoflavone (Cat. # N5757, Sigma)

CYP2C9: 10 mM sulfaphenazol (Cat. # S0758, Sigma)

CYP2C19: 100 mM amitriptyline (Cat. # A8404, Sigma)

CYP2D6: 10 mM quinidine (Cat. # Q3625, Sigma)

CYP3A4: 10 mM ketoconazole (Cat. # K1003, Sigma)

(2) P450-Glo ™ CYP1A2 Screening System (Cat. # V9770, Promega, USA)

(3) P450-Glo ™ CYP2C9 Screening System (Cat. # V9790, Promega, USA)

(4) P450-Glo ™ CYP2C19 Screening System (Cat. # V9880, Promega, USA)

(5) P450-Glo ™ CYP2D6 Screening System (Cat. # V9890, Promega, USA)

(6) P450-Glo ™ CYP3A4 Screening System (Cat. # V9920, Promega, USA)

(7) 96-well Flat Bottom White Polystyrol plate, (Cat. # 3912, Corning, USA)

Was used, specifically, the experimental method,

(1) Dilute the experimental compound (Comparative Example 1) and the positive control stock solution with water to prepare a 40 μM working solution.

(2) A mixed solution containing an enzyme and a substrate is prepared using water and a buffer.

(3) The test compound (Comparative Example 1) / Control / positive control and the mixed solution are each dispensed in 96-well and pre-incubated at 37 ℃ for 10 minutes.

(4) Add NADPH and react at 37 ° C for 20 minutes.

(5) Add detection reagent to terminate the reaction.

(6) After stabilizing at room temperature for 20 minutes, the luminescence signal is measured (the analyzer used is Infinite M1000 Pro (TECAN).

The analyzed results are shown in FIG. 8.

Referring to FIG. 8, the compound of Comparative Example 1 showed significant inhibitory activity (over 50%) in all isoenzymes except CYP2C9, and the compound of Comparative Example 1 was identified as a substance to be considerably drug-interacted compared to the compound of the present invention. It became.

Therefore, the example compound according to the present invention may be provided as a drug that is less affected by drug interactions than the Comparative Example 1 compound and has more safety.

Claims (14)

A compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
[Formula 1]

(In the above formula 1,
X is O, S or NR ¹ ,
Wherein R ¹ is unsubstituted or substituted C _1-10 straight or branched chain alkyl, unsubstituted or substituted C _1-10 straight or branched chain alkoxy, unsubstituted or substituted C _3-7 cycloalkyl , 3 to 7 cyclic unsubstituted or substituted heterocycloalkyl, unsubstituted or substituted C _6-10 aryl containing one or more hetero atoms selected from the group consisting of N, O and S, or N, O And 5 to 10 each ring heteroaryl including one or more hetero atoms selected from the group consisting of S,
Wherein the substituted alkyl, substituted alkoxy, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl or substituted heteroaryl are hydroxy, halogen, -CN, -NO ₂ , C _1-5 May be substituted with straight or branched chain alkyl, C _1-5 straight or branched chain alkoxy;

Is unsubstituted or substituted C _3-7 cycloalkyl, at least one double bond alkenyl cycloalkyl unsubstituted or substituted C _3-7 Al, including, N, O, and one or more heteroatoms selected from the group consisting of S 3 to 7-membered unsubstituted or substituted heterocycloalkyl containing atoms, unsubstituted or substituted C _6-10 aryl, or at least one hetero atom selected from the group consisting of N, O and S 5 to 10 each ring heteroaryl,
Wherein the substituted cycloalkyl, substituted cycloalkenyl, substituted heterocycloalkyl, substituted aryl, or substituted heteroaryl is a straight chain of hydroxy, halogen, -CN, -NO ₂ , C _1-10 Or branched alkyl, C _1-10 straight or branched alkoxy; And

One of W and Y is Br and the other comprises H, unsubstituted or substituted C _6-10 aryl, or a heteroaryl of one member selected from the group consisting of N, O, and S Ring or substituted 5-10 atoms of heteroaryl,
Wherein said substituted aryl, or substituted heteroaryl, is OH, halogen, cyano, nitro, amino, unsubstituted or substituted C _1-10 straight or branched chain alkyl, and unsubstituted or substituted C _1- May be substituted with one or more substituents selected from the group consisting of ₁₀ straight or branched alkoxy,
Here again, the substituted alkyl, or substituted alkoxy, may be substituted with one or more substituents selected from the group consisting of OH, halogen, cyano, nitro, amino, and C _1-3 straight or branched chain alkyl. have).
The method of claim 1,

Is unsubstituted or substituted C _5-6 cycloalkyl, unsubstituted or substituted C _5-6 cycloalkenyl, unsubstituted or substituted phenyl containing one double bond,
Here, the substituted cycloalkyl, substituted cycloalkenyl, or substituted phenyl, at least one selected from the group consisting of straight chain or branched chain alkoxy of alkyl and C _1-5 straight or branched-chain C _1-5 Compounds, stereoisomers thereof, or pharmaceutically acceptable salts thereof, which may be substituted with substituents.
The method of claim 1,
X is O; And

Is

,

,

,

,

,

,

,

,

,

,

,

or

Compounds, stereoisomers thereof, or pharmaceutically acceptable salts thereof, characterized in that.
The method of claim 1,
The compound represented by Formula 1 is any one selected from the following compound group, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
(1) (Z) -3- (bromo (phenyl) methylene) isobenzofuran-1 (3H) -one;
(2) (Z) -3- (bromo (thiophen-2-yl) methylene) isobenzofuran-1 (3H) -one;
(3) (Z) -3- (bromo (furan-2-yl) methylene) isobenzofuran-1 (3H) -one;
(4) (Z) -3- (bromo (4-chlorophenyl) methylene) isobenzofuran-1 (3H) -one;
(5) (Z) -3- (bromo (4-fluorophenyl) methylene) isobenzofuran-1 (3H) -one;
(6) (Z) -3- (bromo (4- (trifluoromethyl) phenyl) methylene) isobenzofuran-1 (3H) -one;
(7) (Z) -3- (bromo (pyridin-2-yl) methylene) isobenzofuran-1 (3H) -one;
(8) (E) -3- (bromo (pyridin-2-yl) methylene) isobenzofuran-1 (3H) -one;
(9) (Z) -3- (bromo (pyridin-3-yl) methylene) isobenzofuran-1 (3H) -one;
(10) (E) -3- (bromo (pyridin-3-yl) methylene) isobenzofuran-1 (3H) -one;
(11) (Z) -3- (bromo (pyrimidin-5-yl) methylene) isobenzofuran-1 (3H) -one; And
(12) (E) -3- (bromo (pyrimidin-5-yl) methylene) isobenzofuran-1 (3H) -one.
As shown in Scheme 1 below,
Preparing a compound represented by Chemical Formula 2 from the compound represented by Chemical Formula 3; And
A method for preparing a compound represented by Formula 1 of claim 1, comprising: preparing a compound represented by Formula 1 from the compound represented by Formula 2 prepared in the step:
Scheme 1

(In the above Reaction Scheme 1,

, W, X, and Y are as defined in formula (1) of claim 1).
A pharmaceutical composition for the prevention or treatment of oral diseases, comprising a compound represented by the formula (1) of claim 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
The method of claim 6,
The compound is a pharmaceutical composition for the prevention or treatment of oral diseases, characterized in that the quorum sensing of bacteria, or inhibiting the biofilm formation of bacteria.
Toothpaste composition for the prevention or improvement of oral diseases comprising the compound of claim 1, its stereoisomers, or pharmaceutically acceptable salts thereof as an active ingredient.
A gargle composition for preventing or ameliorating oral diseases, comprising the compound of claim 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
Health functional food composition for the prevention or improvement of oral diseases containing a compound represented by the formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
A pharmaceutical composition for the prophylaxis or treatment of an inflammatory disease containing a compound represented by the formula (1) of claim 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
The method of claim 11,
The inflammatory disease is a pharmaceutical composition, characterized in that the inflammation of the mucous (mucous) and mucocutaneousus tissue of the oral cavity (dental pulp), or oral bacterial infectious inflammation.
An antibiotic composition containing the compound represented by the formula (1) of claim 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
The method of claim 13,
The antibiotic composition is an antibiotic composition, characterized in that the antibacterial or antifungal agent.

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