CN108101796B - Tertiary amine compound and application thereof - Google Patents

Abstract

The invention provides a novel tertiary amine compound, which can reduce the pH value of the environment under the action of acid-producing bacteria, plays an antibacterial role, has the pH sensitive characteristic and solves the problem of dysbacteriosis caused by lack of selectivity of the traditional antibacterial agent.

Description

Tertiary amine compound and application thereof

Technical Field

The invention relates to a novel tertiary amine compound and application thereof.

Background

Caries is one of the most common chronic diseases, and as a plaque disease, the occurrence of caries is closely related to plaque biofilm in the oral cavity. The bacterial plaque biomembrane is planted on the surface of the acquired dental membrane, carbohydrate is utilized to generate extracellular matrix, the extracellular matrix further grows and matures in the extracellular matrix, the pH value of a microenvironment is gradually reduced, and the microbial species are transferred to acid-producing bacteria and acid-resisting bacteria, so that the tooth is demineralized, and dental caries is generated.

For years, a great deal of related researches aiming at inhibiting bacterial plaque biological membranes, such as chlorhexidine, hydrogen peroxide, fluorine preparations, nano materials and the like, have obvious effects in the prevention and treatment researches of caries in the forms of solution, suspension, gel and the like; however, there are problems such as a significant decrease in the effect of chlorhexidine, hydrogen peroxide, and the like on the film-forming microorganisms. Once the resin edge is slightly leaked after the filling treatment is finished for a carious patient, dental plaques are easy to gather to generate secondary caries, and the effect of the medicine on preventing and treating the secondary caries is limited. In recent years, many studies have been made on modified resin filling materials, and for example, quaternary ammonium salt modified resin filling material systems have a remarkable effect on biofilms adhered to the surfaces of resins, teeth and adhesive surfaces.

However, a large amount of oral resident bacteria exist in the oral cavity, which is closely related to the maintenance of oral health and oral micro-ecological balance, and the research materials have ideal results in the generation of bacteria resistance, caries prevention or secondary caries, but lack selectivity on the action of microorganisms, and have the possibility of causing dysbacteriosis, thereby generating new problems. Nowadays, many researches on pH-sensitive nanomaterials for caries prevention are being conducted, and having antibacterial activity in a low pH state is one of effective methods for solving the above problems. However, the materials which can modify the resin filling material system and have pH sensitive characteristics are still not studied.

Disclosure of Invention

Based on the above problems, the present invention provides a novel tertiary amine compound which can be used against oral acid-producing bacteria.

Specifically, the invention provides a tertiary amine compound shown as a formula I:

wherein R1 and R2 are independently selected from H, C1-C4 alkyl;

r3 is selected from C1-C4 alkyl;

n is an integer from 12 to 16.

The C1-C4 alkyl group in the present invention includes both straight chain alkyl groups and branched chain alkyl groups. It may be a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group or the like.

Further, R1 and R2 are each independently selected from methyl or ethyl. In one embodiment of the invention, R1 and R2 are each independently selected from methyl.

Further, R3 is selected from methyl or ethyl. In one embodiment of the invention, R3 is selected from methyl.

Further, n is selected from integers of 13-15, such as 13, 14, 15. In one embodiment of the invention, n is 14.

The following experimental studies were carried out on the compounds in the present invention to illustrate the beneficial effects of the compounds:

1. determination of Minimum Inhibitory Concentration (MIC) at different pH

Dissolving 16-chain tertiary amine and 16-chain quaternary ammonium salt in BHI culture medium (pH 5, pH 7.4) with different pH values and Mcbain culture medium with neutral buffer system (pH 7), placing in 96-well plate, gradient diluting, and culturing 24H in BHI culture medium (37 deg.C, 90% N)₂，5％CO₂，5％H₂) The streptococcus mutans suspension is added into a culture medium of a gradient dilution antibacterial medicament and is added into the culture medium (the concentration is 10)⁷CFU/ml) 24H (37 ℃, 90% N)₂，5％CO₂，5％H₂) MIC was determined based on turbidity of the medium in 96-well plates.

2. Preparation of experimental sample and biofilm culture of streptococcus mutans

The experimental groups were adhesive group (control group), 5% 16-chain quaternary ammonium salt + adhesive group (5% 16-chain quaternary ammonium salt group), and 5% 16-chain tertiary amine + adhesive group (5% 16-chain tertiary amine group), wherein the 16-chain tertiary amine was methyl hexadecylamino ethyl methacrylate.

A circular resin sheet with a diameter of 1.2cm is prepared, and is photo-cured after a thin layer of adhesive is uniformly coated on the resin sheet.

After ethylene oxide sterilization at 37 ℃ each group of resin chips was placed in a 24-well plate and Streptococcus mutans (Streptococcus mutans UA159) was cultured on a thin layer adhesive. Anaerobic environment (90% N) in culture medium of Streptococcus mutans and BHI₂，5％CO₂，5％H₂) After 24H culture, the mixture is mixed in equal proportion in BHI containing 1% sucrose and Mcbain culture medium with neutral buffer system, and then added into the above 24-well plate (the concentration of three strains are all 10)⁷CFUs/ml), 2ml per well, 48H anaerobically.

3. Thiazole blue colorimetric method (MTT method) detection

Transferring the resin sheets (6 in each group) with 48H biological membranes into a new 24-well plate, respectively putting the resin sheets into 1ml of MTT dye (0.5mg/ml, dissolved in PBS), carrying out anaerobic culture at 37 ℃ for 1H, transferring the resin sheets into the new 24-well plate, respectively adding 1ml of dimethyl sulfoxide (DMSO), preventing and treating at room temperature for 20min in a dark place, taking 200ul of each well, putting the well into a 96-well plate, and measuring the degree under the OD value of 540 nm.

4. Lactic acid test

The resin sheets (6 per group) cultured with 48H biofilm were transferred to a new 24-well plate, and 1.5ml of Buffered Peptone Water (BPW) containing 0.2% sucrose was placed therein, and the amount of lactic acid produced was measured by the lactic acid dehydrogenase method after anaerobic culture at 37 ℃ for 3H, and the OD value was 340 nm.

The test results are shown in table 1, fig. 1 and fig. 2.

TABLE 1

As shown in the above table, in the acidic medium, the antibacterial effect of the 16-chain tertiary amine is obviously enhanced, while the antibacterial effect of the 16-chain quaternary ammonium salt is not changed greatly, and the antibacterial potential of the tertiary amine in pH sensitivity can be seen.

The figure shows the effect of tertiary amines of the invention on a 48-hour Streptococcus mutans biofilm in different media containing the same concentration of quaternary ammonium salt (DMAHDM). As can be seen from the figure, the antibacterial effects of the quaternary ammonium salt and the tertiary amine at the same concentration in the BHI medium (pH 4.1 after the cultivation) on the 48-hour biofilm were similar. While in McBain medium (pH 6.1 after cultivation) tertiary amines have no significant antibacterial effect on biofilms. The above results also demonstrate that the tertiary amine compounds provided by the present invention are pH sensitive, whereas DMAHDM does not.

Based on the activity experiment, the invention also provides the application of the compound in preparing medicines or materials with antibacterial or antibacterial biological membrane activity.

Further, the bacteria are acid-producing bacteria.

Wherein the bacterium is Streptococcus mutans.

The invention also provides the use of the above compounds for the preparation of a medicament, prodrug or material having antibacterial or antibacterial biofilm activity under acidic conditions.

The invention also provides an antibacterial drug or material which takes the compound as an active ingredient.

The tertiary amine compound synthesized by the invention has the advantages of reducing the pH value of the environment under the action of acid-producing bacteria, playing an antibacterial role, having a pH sensitive characteristic, and solving the problem of flora imbalance caused by lack of selectivity of a quaternary ammonium salt antibacterial agent compared with DMAHDM.

The medicine or the material can be used for biomedical materials (such as dentistry) and can also be used in other fields. In one embodiment of the invention, the material is a biomedical material, in particular a dental material.

The dental material is also called as oral material, is one of biomedical materials, can be used in oral biological environment, and can be used for repairing defective teeth or replacing defective and missing dentition, restoring anatomical form, function and beauty, and various materials used in medical treatment such as oral prevention and health care, correction of deformity and the like.

In the using process, in order to meet the actual using requirements, the adhesive can be combined with other auxiliary materials, such as a dental adhesive and the like.

The prodrug of the present invention is a compound having no or little activity in vitro and exerting pharmacological effects by being converted into an active drug in vivo by an enzyme or a nonenzymatic enzyme. The tertiary amine compound provided by the invention has no antibacterial activity under normal neutral environment, and can generate antibacterial activity under acidic condition or in the presence of acid-producing bacteria, so that the tertiary amine compound can also be used as a prodrug.

Drawings

FIG. 1 results in BHI Medium

FIG. 2 results in McBain Medium

FIG. 3 NMR spectra of methylhexadecylaminoethyl methacrylate

Detailed Description

EXAMPLE 1 Synthesis of methylhexadecylaminoethyl methacrylate (Pro B, also referred to herein as 16-chain tertiary amine)

(1) Synthesis of compound B2: in a 250ml round bottom flask, the compounds bromohexadecane (0.1mol,25g,25ml) and compound A2 (N-methylethanolamine) (5.0eq,0.5mol,32g,34ml) were added, and 80ml isopropanol was added as a solvent, and the mixture was refluxed at 85 ℃ for 8-10h, and a large amount of product was found to be produced by Thin Layer Chromatography (TLC). The system was cooled to room temperature, diluted into 150mL of ether and washed with water (3 x 150mL), saturated brine (2 x 150mL), the organic phase collected and purified over anhydrous Na₂SO₄Drying and removal of the solvent under reduced pressure gave B as a colorless oil (24g, up to 99% yield).

(2) Synthesis of Compound Pro B2(0.1mol,24g) and methyl methacrylate (3.0eq,0.3mol,31.8ml) were added to a 100ml round bottom flask and stirred well. Catalyst CAA (0.4 mol%, 0.107g,) and polymerization inhibitor p-hydroxyanisole (2 mol%, 100mg) were added and stirred well. The reaction system is reacted at the temperature of 100 ℃ and 110 ℃, and a little methyl methacrylate can be distilled under the normal pressure distillation device. After the reaction time of 12h, the colorless system changed to yellow and gradually deepened, the catalyst CAA (0.6 mol%, 150mg) was added, the reaction was continued for 12h, and after the system returned to room temperature, column chromatography was performed to obtain methyl hexadecylamino ethyl methacrylate (Pro B) (4.2g, 14% yield) as a yellow transparent oily substance.

EXAMPLE 2 dental antibacterial Material

A proper amount of methyl hexadecyl amino ethyl methacrylate is taken and mixed with the adhesive uniformly, and then the mixture can be used as the dental antibacterial material.

The dental antibacterial material can be a product formed by mixing methyl hexadecyl aminoethyl methacrylate with an adhesive or a separated product.

Claims (6)

1. A tertiary amine compound represented by formula I:

wherein R1 and R2 are respectively selected from methyl; r3 is selected from methyl; n is selected from 14.

2. Use of a compound according to claim 1 for the preparation of a medicament or material having antibacterial activity.

3. Use of a compound according to claim 1 for the preparation of a medicament or material having antibacterial biofilm activity.

4. Use according to claim 2 or 3, characterized in that: the bacteria are acid-producing bacteria.

5. Use according to claim 4, characterized in that: the bacteria are streptococcus mutans.

6. An antibacterial drug or material, characterized in that: which comprises the compound according to claim 1 as an active ingredient.

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