WO2021071136A2 - Nouvel agent de conservation, et son procédé de production - Google Patents

Nouvel agent de conservation, et son procédé de production Download PDF

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WO2021071136A2
WO2021071136A2 PCT/KR2020/012784 KR2020012784W WO2021071136A2 WO 2021071136 A2 WO2021071136 A2 WO 2021071136A2 KR 2020012784 W KR2020012784 W KR 2020012784W WO 2021071136 A2 WO2021071136 A2 WO 2021071136A2
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preservative
reaction
formula
alkyl
vanillin
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PCT/KR2020/012784
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Korean (ko)
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WO2021071136A3 (fr
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원기훈
이수진
김성준
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동국대학교 산학협력단
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Priority claimed from KR1020200122337A external-priority patent/KR102419005B1/ko
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids

Definitions

  • the present invention relates to a novel preservative and a method of preparing the same.
  • Preservatives refer to raw materials/materials that play a role in inhibiting or reducing the growth of microorganisms during storage and use of the product. Preservatives are widely used not only during the shelf life of cosmetics, but also to prevent changes in physicochemically such as decay and deterioration due to contamination by microorganisms such as bacteria and fungi while consumers use the product. Preservatives are used not only in cosmetics, but also in many consumer goods such as food and medicine. Representative preservatives used in cosmetics include paraben and phenoxyethanol, but these preservatives are currently out of demand from consumers due to problems such as toxicity, skin irritation, and allergies.
  • An object of the present invention is to provide a novel preservative.
  • An object of the present invention is to provide a method for producing a novel preservative.
  • a preservative comprising a compound of Formula 1 below:
  • R 1 is C1 to C6 alkyl
  • R 2 is hydrogen or C1 to C6 alkyl.
  • R 1 is C1 to C3 alkyl
  • R 2 is hydrogen or C1 to C3 alkyl
  • 3,4-dihydroxybenzyl butyrate (3,4-dihydroxybenzyl butyrate) is at least one selected from the group consisting of, a preservative.
  • preservative according to the above 1, wherein the preservative exhibits antimicrobial activity against at least one selected from the group consisting of Escherichia, Staphylococcus, Candida, and Aspergillus.
  • preservative is at least one of a cosmetic preservative, a food preservative, and a pharmaceutical preservative.
  • a method for preparing a preservative comprising reacting a compound of the following formula 2 or a compound of the following formula 3, and a compound of the following formula 4 in the presence of a lipase:
  • R 3 is C1 to C6 alkyl
  • R 6 is C1 to C6 alkyl
  • R 4 is C1 to C6 alkyl
  • R 5 is hydrogen or C1 to C6 alkyl
  • R 3 is C1 to C3 alkyl
  • R 4 is C1 to C3 alkyl
  • R 5 is hydrogen or C1 to C3 alkyl
  • R 6 is C1 to C3 Alkyl phosphorus
  • a cosmetic composition comprising the preservative of any one of the above 1 to 5.
  • the preservative of the present invention exhibits excellent antibacterial activity against gram-positive bacteria, gram-negative bacteria, yeast, and mold.
  • the manufacturing method of the present invention is to enzymatically convert natural phenolic compounds obtained from biomass, which is a renewable resource, to prepare a preservative.With a small amount of enzyme, a high reaction conversion rate even under mild reaction conditions at room temperature and pressure within a short time. Represents.
  • Figure 2 shows the HPLC analysis results and reaction conversion of the enzymatic esterification reaction of vanillic acid and n-propanol.
  • Figure 10 is a comparison of the reaction conversion rates of the enzymatic transesterification of vanillin alcohol and ethyl acetate, the enzymatic transesterification of vanillin alcohol and ethyl propionate, and the enzymatic transesterification of vanillin alcohol and ethyl butyrate. Show one graph.
  • FIG. 11 shows the reaction scheme of the enzymatic transesterification reaction of vanillin alcohol and ethyl butyrate.
  • Fig. 16 shows the results of evaluating the cytotoxicity of vanillin propionate through L/D fluorescence staining (target cells: keratinocytes, green: living cells, red: dead cells).
  • Fig. 17 shows the results of evaluating the cytotoxicity of vanillin propionate through L/D fluorescence staining (target cells: dermal fibroblasts, green: living cells, red: dead cells).
  • 19 shows the reaction conversion rate of the enzymatic transesterification reaction of vanillin alcohol and triacetin.
  • Figure 20 shows the reaction scheme of the enzymatic transesterification reaction of 3,4-dihydroxybenzyl alcohol and ethyl propionate.
  • Figure 21 shows the reaction conversion rate of the enzymatic transesterification reaction of 3,4-dihydroxybenzyl alcohol and ethyl propionate.
  • the present invention provides a preservative comprising a compound of Formula 1:
  • R 1 may be C1 to C6 alkyl. Specifically, R 1 may be C1 to C3 alkyl.
  • R- 2 may be hydrogen or C1 to C6 alkyl. Specifically, R 2 may be hydrogen or C1 to C3 alkyl.
  • Alkyl can be straight or branched chain alkyl.
  • preservative refers to a substance that is customarily added to a composition to preserve the composition from substances causing contamination.
  • the compound of Formula 1 is 4-hydroxy-3-methoxybenzyl propionate
  • It may be at least one selected from the group consisting of.
  • 4-hydroxy-3-methoxybenzyl propionate; 4-hydroxy-3-methoxybenzyl acetate; 4-hydroxy-3-methoxybenzyl butyrate; 3,4-dihydroxybenzyl propionate; 3,4-dihydroxybenzyl acetate and 3,4-dihydroxybenzyl butyrate may exhibit antimicrobial and/or antibacterial and/or antifungal effects.
  • 4-hydroxy-3-methoxybenzyl propionate; 4-hydroxy-3-methoxybenzyl acetate; 4-hydroxy-3-methoxybenzyl butyrate; 3,4-dihydroxybenzyl propionate; 3,4-dihydroxybenzyl acetate and 3,4-dihydroxybenzyl butyrate may each exhibit antibacterial effects against different bacteria.
  • the preservative of the present invention may exhibit antibacterial activity against at least one of bacteria, fungi and yeast.
  • the preservatives of the present invention can be used as antimicrobial and/or antibacterial and/or antifungal agents.
  • Bacteria may be any of Enterococcus (Enterococcus) genus, the Escherichia (Escherichia) genus, Staphylococcus (Staphylococcus) genus Pseudomonas (Pseudomonas) in and Burke holde Liao at least one of the inside (Burkholderia), but is not limited thereto.
  • bacteria are Enterococcus faecalis , Escherichia coli , Staphylococcus aureus , Pseudomonas aeruginosa , Burkholderia sepacia ( Burkholderia cepacia ).
  • the fungus may be of the genus Aspergillus , but is not limited thereto.
  • the fungus may be Aspergillus niger.
  • Yeast may be of the genus Candida , but is not limited thereto.
  • the yeast may be Candida albicans.
  • the preservative of the present invention may be at least one of a cosmetic preservative, a food preservative, and a pharmaceutical preservative.
  • the preservative of the present invention may be included in a cosmetic composition, a pharmaceutical composition, or a food composition to exhibit a preservative power capable of preserving the composition from contamination by microorganisms.
  • preservation power refers to the defense power that can prevent the product from deteriorating and maintain its original state.
  • the preservative of the present invention may not impair the texture, color, or odor of a cosmetic composition, pharmaceutical composition or food composition containing the preservative.
  • the preservative of the present invention may not contain known chemical preservatives other than the compound of Formula 1 above.
  • the preservative of the present invention may not contain chemical preservatives such as parabens and phenoxyethanol, and in that case, problems such as toxicity, skin irritation, and allergy caused by the use of chemical preservatives can be solved.
  • the preservative of the present invention may exhibit the preservative power or antibacterial power similar to or superior to the preservative power or antibacterial power of the conventional chemical preservatives of parabens.
  • the present invention provides a method for preparing a preservative.
  • the method for preparing a preservative of the present invention may include reacting a compound of Formula 2 or Formula 3 and a compound of Formula 4 in the presence of a lipase (hereinafter, a substrate reaction step).
  • R 3 may be C1 to C6 alkyl. Specifically, R 3 may be C1 to C3 alkyl.
  • R 6 may be C1 to C6 alkyl. Specifically, R 6 may be C1 to C3 alkyl.
  • R 4 may be C1 to C6 alkyl. Specifically, R 4 may be C1 to C3 alkyl.
  • R 5 may be hydrogen or C1 to C6 alkyl. Specifically, R 5 may be C1 to C3 alkyl.
  • the compound of Formula 4 may be a phenolic alcohol.
  • the compound of Formula 4 may be vanillin alcohol or 3,4-dihydroxybenzyl alcohol.
  • Vanillin alcohol or 3,4-dihydroxybenzyl alcohol may be naturally obtained or synthesized. Vanillin alcohol may be produced through a reduction reaction of vanillin, a natural phenolic compound.
  • the compound of Formula 2 and the compound of Formula 3 are used as an acyl donor, and the compound of Formula 4 is used as an acyl acceptor.
  • glycerol is produced as a by-product.
  • glycerol is a representative material used as a moisturizing component of cosmetics
  • the substrate reaction step may be performed under no solvent. This is because a separate organic solvent is not required since the compound of Formula 2 or the compound of Formula 3 is a liquid substrate.
  • the lipase used in the substrate reaction step can be used without limitation, known in the art.
  • the lipase may be Candida antarctica lipase (CAL), Rhizomucor miehei lipase (RML), Rhizopus javanicus lipase (RJL), Rhizopus delemar lipase (RDL).
  • CAL Candida antarctica lipase
  • RML Rhizomucor miehei lipase
  • RJL Rhizopus javanicus lipase
  • RDL Rhizopus delemar lipase
  • the concentration of lipase used in the substrate reaction step may be 50 U/mL or less, for example, 50 U/mL or less, 45 U/mL or less, 40 U/mL or less, 35 U/mL or less, 30 U/mL or less, It may be 25 U/mL or less, 20 U/mL or less, 15 U/mL or less, 10 U/mL or less, or 5 U/mL or less.
  • the lower limit of the lipase concentration used in the substrate reaction step is 0.1 U/mL, 0.5 U/mL, 1 U/mL, 1.5 U/mL, 2 U/mL, 2.5 U/mL, 3 U/mL, 3.5 U/mL. , 4 U/mL or 4.5 U/mL.
  • reaction conversion is high even in the presence of lipase at a concentration of 50 U/mL or less.
  • the reaction conversion rate is not high even if the reaction is carried out in the presence of a lipase of about 8 times higher than the above concentration.
  • the substrate reaction step may be performed under a temperature condition of 70° C. or less.
  • the substrate reaction step is 70°C or less, 65°C or less, 60°C or less, 55°C or less, 50°C or less, 45°C or less, 40°C or less, 35°C or less, 30°C or less, 25°C or less, 20°C or less , It may be performed under a temperature condition of 15°C or less, 10°C or less, or 5°C or less.
  • the lower limit of the temperature condition of the substrate reaction step may be 5°C, 10°C, 15°C, 20°C, 25°C, 30°C or 35°C.
  • the reaction temperature conditions may vary depending on the type or concentration of the substrate used in the substrate reaction step, the reaction conversion rate is high even at a low temperature condition of 70° C. or less when the compound of Formula 4 is used as a reactant.
  • the reaction conversion rate is not high even if the reaction is carried out under a high temperature condition exceeding 70°C.
  • the reaction may be performed for a period of 10 hours or less.
  • the substrate reaction step is performed for a time period of 10 hours or less, 9 hours or less, 8 hours or less, 7 hours or less, 6 hours or less, 5 hours or less, 4 hours or less, 3 hours or less, 2 hours or less, or 1 hour or less. It can be.
  • the lower limit of the reaction execution time in the substrate reaction step may be 30 minutes, 1 hour, or 1 hour 30 minutes.
  • the reaction conversion rate is high even if the reaction is performed for a short time of 10 hours or less.
  • the reaction conversion rate is not high even if the reaction is performed for more than 10 hours.
  • the concentration of the compound of Formula 4 used in the substrate reaction step may be 10 to 300 mM, for example, the concentration of vanillin alcohol is 10 to 300 mM, 20 to 290 mM, 30 to 280 mM, 40 to 270 mM, 50 to 260 mM, 60 to 250 mM, 70 to 240 mM, 80 to 230 mM, 90 to 220 mM, 100 to 210 mM, 110 to 200 mM, 120 to 190 mM, 130 to 180 mM, 140 to 170 mM, 150 to It may be 160 mM, but is not limited thereto.
  • the substrate reaction step may be to react the compound of Formula 2 and the compound of Formula 4 in a molar ratio of 1 to 200:1.
  • the substrate reaction step may be to react the compound of Formula 2 and the compound of Formula 4 in a molar ratio of 1 to 200:1, 10 to 100:1, 20 to 50:1, 30 to 50:1, Specifically, it may be reacted at a molar ratio of 30 to 50:1.
  • the substrate reaction step may be to react the compound of Formula 3 and the compound of Formula 4 in a 1: 1 to 3 molar ratio.
  • the substrate reaction step may be to react the compound of Formula 3 and the compound of Formula 4 in a 1: 1 to 3 molar ratio or 1: 1.5 to 2.5 molar ratio.
  • the present invention can provide a cosmetic composition comprising the preservative described above.
  • the cosmetic composition of the present invention exhibits excellent preservation power and safety by including the preservative described above.
  • the content of the preservative contained in the cosmetic composition of the present invention is not particularly limited as long as the desired preservative power can be secured, and those skilled in the art can determine the appropriate content according to the degree of desired preservative power.
  • the preservative of the present invention may be included in an amount of 0.01 to 20% by weight based on the total cosmetic composition.
  • the preservative of the present invention may be included in an amount of 0.01 to 20% by weight, 0.1 to 10% by weight, 0.1 to 5% by weight, or 0.1 to 2% by weight based on the total cosmetic composition.
  • the cosmetic composition may be any known formulation, for example, skin, emulsion, cream, sun cream, foundation, essence, gel, pack, mask pack, foam cleanser, body cleanser, softening lotion, eyeliner, shampoo, conditioner, soap , Hairdressing agent, wool agent, hair cream, hair styling gel, lubricant, toothpaste, and may be one formulation selected from the group consisting of wet tissues, but is not limited thereto.
  • the cosmetic composition of the present invention may further contain known additives within a range that does not impair the effect of the preservative of the present invention depending on the formulation.
  • the cosmetic composition of the present invention further comprises an additive selected from the group consisting of a carrier, an emulsifier, a moisturizer, a skin conditioning agent, a surfactant, a chelating agent, an antioxidant, a fungicide, a stabilizer, and any combination thereof. I can.
  • Carrier is animal fiber, plant fiber, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, lactose, silica, aluminum hydroxide, calcium silicate, polyamide powder , Water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, liquid diluent, ethoxylated isostearyl alcohol, polyoxy Ethylene sorbitol ester, polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid mono
  • the emulsifier may be at least one selected from the group consisting of liquid paraffin, cetyloctanoate, and stearic acid, but is not limited thereto.
  • the moisturizing agent may be at least one selected from the group consisting of glycerin and glyceryl stearate, but is not limited thereto.
  • the skin conditioning agent may be at least one selected from the group consisting of saliclomethicone and dimethicone, but is not limited thereto.
  • the surfactant may be at least one selected from the group consisting of cetostearyl alcohol, triethanolamine, and carboxyvinyl polymer, but is not limited thereto.
  • the chelating agent may be at least one selected from the group consisting of sodium ethylenediaminetetraacetate (EDTA), ⁇ -hydroxy fatty acid, lactoferrin, ⁇ -hydroxy acid, citric acid, lactic acid, malic acid, bilirubin, and biliverdin. However, it is not limited thereto.
  • EDTA sodium ethylenediaminetetraacetate
  • lactoferrin ⁇ -hydroxy acid
  • citric acid lactic acid
  • malic acid bilirubin
  • biliverdin biliverdin
  • the antioxidant may be at least one selected from the group consisting of butylhydroxyanisole, dibutylhydroxytoluene, and propyl gallate, but is not limited thereto.
  • the cosmetic composition of the present invention may further include at least one of a fat or oil component, an emollient agent, an organic pigment, an inorganic pigment, an organic powder, an ultraviolet absorber, a pH adjuster, an alcohol, a colorant, a fragrance, a blood circulation accelerator, a cooling agent, and a limiting agent.
  • a fat or oil component an emollient agent, an organic pigment, an inorganic pigment, an organic powder, an ultraviolet absorber, a pH adjuster, an alcohol, a colorant, a fragrance, a blood circulation accelerator, a cooling agent, and a limiting agent.
  • fractionated by reaction time, diluted 20 times, and analyzed by HPLC under 275 nm conditions see FIG. 2). As a result, it showed a very low reaction conversion rate (conversion) within 5% of the case.
  • Example 1 Preparation of a preservative using vanillin alcohol and short chain ester
  • the short-chain ester includes ethyl acetate, ethyl propionate, and ethyl butyrate used in the following reaction, and is a short-chain hydrocarbon (for example, in the ester group (-COO-)). It refers to a compound to which a chain having C1-6 carbon atoms) is bonded.
  • An enzymatic transesterification reaction was performed using vanillin alcohol, a natural phenolic compound, as an acyl acceptor substrate and ethyl propionate as an acyl donor substrate. At this time, the reaction was carried out in a solvent-free system by replacing the solvent with ethyl propionate as a liquid (see FIG. 3).
  • liquid ethyl propionate (total volume 2 mL) and solid vanillin alcohol (50, 100, 200 mM) were prepared and completely dissolved in a 20 mL glass vial.
  • the molar ratio of ethyl propionate and 50mM vanillin alcohol was 173:1
  • the molar ratio of ethyl propionate and 100mM vanillin alcohol was 87:1
  • the molar ratio of ethyl propionate and 200mM vanillin alcohol was 43:1.
  • HPLC High Performance Liquid Chromatography
  • the elution time conditions are as follows; 0-2 min: A 50%-B 50 %, 2-7 min: A 90%-B 10 %, 7-10 min: A 90%-B 10 %, 10-15 min: A 50%-B 50 %. All samples were detected by Diode Array Detector (DAD) at 280 nm.
  • DAD Diode Array Detector
  • the conversion rate could be greatly improved by using vanillin alcohol instead of vanillic acid as a reaction substrate.
  • reaction stock solution was transferred to a 15 mL centrifuge tube, and then molecular sieves (4 ⁇ ) and enzyme were separated and removed through centrifugation at 10,000 rpm for 10 to 15 minutes. Evaporation was performed for 30-40 minutes in a vacuum rotary evaporator at 60°C to remove the liquid ethyl propionate remaining without reaction and to proceed with concentration.
  • the recovered product was stored in a vacuum desiccator for 2 to 3 days, and then ethyl propionate was completely removed.
  • the chemical structure of the finally obtained material was analyzed through 400 MHz H 1 -NMR, and the analysis result was consistent with the chemical structure of 4-hydroxy-3-methoxybenzyl propionate (vanillin propionate) (see FIG. 7 ). ).
  • the enzymatic transesterification reaction was carried out for 5 hours under the conditions of ethyl acetate with vanillin alcohol 100 mM, enzyme 10 U/mL, molecular sieves 20 mg/mL, 35 °C, 200 rpm (see Fig.8: vanillin alcohol and Enzymatic transesterification reaction with ethyl acetate as a reactant).
  • vanillin alcohol As a reaction product, 4-hydroxy-3-methoxybenzyl acetate (vanillin acetate) was produced.
  • the above-described reaction between ethyl acetate and vanillin alcohol showed a high reaction conversion rate of 90% or more (see FIG. 10).
  • reaction stock solution was transferred to a 15 mL centrifuge tube, and then molecular sieves (4 ⁇ ) and enzyme were separated and removed through centrifugation at 10,000 rpm for 10 to 15 minutes. Evaporation was performed for 30 to 40 minutes in a vacuum rotary evaporator at 50 °C to remove the remaining liquid ethyl acetate without reaction and to proceed with concentration. The recovered product was completely dried after storage for 2 to 3 days in a vacuum desiccator. As a result of analyzing the chemical structure of the finally obtained material through 400 MHz H 1 -NMR, it was consistent with the chemical structure of 4-hydroxy-3-methoxybenzyl acetate (see FIG. 9).
  • an enzymatic transesterification reaction was performed for 5 hours under the conditions of ethyl butyrate with vanillin alcohol 100 mM, enzyme 10 U/mL, molecular sieves 20 mg/mL, 35 °C, 200 rpm (see FIG. 11).
  • Enzymatic transesterification reaction with vanillin alcohol and ethyl butyrate as reactants As a result, 4-hydroxy-3-methoxybenzyl butyrate (vanillin butyrate) was produced.
  • the above-described reaction between ethyl butyrate and vanillin alcohol showed a high reaction conversion rate of 90% or more (see FIG. 10).
  • a paper disc diffusion assay was performed to determine the antimicrobial efficacy of the vanillin ester (vanillin propionate, vanillin acetate, and vanillin butyrate) prepared by the above method.
  • Microorganisms used to evaluate antibacterial efficacy are shown in Table 1 below.
  • microorganisms in Table 1 are pathogenic microorganisms, and are representative microorganisms used when evaluating the efficacy of cosmetic preservative components.
  • a disc obtained by soaking a certain amount (20 ⁇ L) of a sample was placed on a plate coated with pathogenic bacteria and cultured.
  • a clear zone is formed because the treated sample spreads to a disc (diameter: 8 mm) and inhibits the growth of bacteria.
  • This transparent ring is referred to as the microbial growth inhibition zone and is also referred to as the inhibition zone.
  • the antibacterial effect is evaluated by measuring the diameter (unit: mm) of the transparent ring created on the disc, and it can be interpreted that the larger the diameter of the inhibition zone is, the better the antibacterial effect is (see FIG. 13(A)).
  • the antibacterial efficacy was evaluated at 2 wt% for all samples.
  • DMSO was used as a solvent to make a 2 wt% sample stock solution.
  • a vanillin ester such as vanillin propionate and vanillin butyrate prepared by the 1.
  • Preparation method of Example 1 was used, and as a control, the existing preservatives methyl paraben, ethyl paraben, propyl paraben, butyl paraben, phenoxyethanol , 1,2-hexanediol and vanillin alcohol as a substrate material before the reaction were used.
  • vanillin ester showed antibacterial efficacy, and in particular, vanillin butyrate showed excellent antibacterial efficacy comparable to butyl paraben in all of E. coli , S. aureus and C. albicans strains (see FIG. 13(B)). , ND: not detected). If various kinds of vanillin ester are used together as a preservative, it can be expected to produce synergistic effects.
  • MIC Minimum Inhibitory Concentration
  • the MIC of the vanillin propionate prepared by the above method was confirmed, and compared with the MIC values of the existing preservatives (methyl paraben, ethyl paraben, propyl paraben, phenoxyethanol, 1,2-hexanediol).
  • the vanillin ester showed significantly lower MIC values compared to phenoxyethanol and 1,2-hexanediol.
  • it exhibited similar or lower MIC values compared to the parabens compound (see FIG. 14).
  • the types of bacteria and antibacterial activity may vary according to the carbon length of the substituent.
  • vanillin acetate, vanillin propionate, and vanillin butyrate obtained using various types of acyl donor substrates is expected to exhibit more excellent antibacterial efficacy against various bacteria.
  • Cytotoxicity evaluation was performed on the vanillin ester and the positive control parabens. Cytotoxicity evaluation was performed on human keratinocytes (HaCaT cells) and human dermal fibroblasts (HDFs), which are cells that make up the skin.If toxicity to skin cells is low, it is possible to commercialize it in the cosmetic industry. It becomes higher. Vanillin propionate; Propyl parabens similar in structure to this; Ethyl paraben, which has similar antimicrobial efficacy, was used as a sample for toxicity evaluation, and WST-1 Assay and L/D fluorescence staining were performed as a toxicity evaluation method.
  • HaCaT cells human keratinocytes
  • HDFs human dermal fibroblasts
  • the metabolic activity of mitochondria in cells was quantitatively measured through WST-1 Assay.
  • ethyl paraben and propyl paraben did not show cytotoxicity up to 0.5 mM, whereas vanillin propionate did not show cytotoxicity up to a higher concentration of 1.5 mM.
  • propyl parabens all killed cells at a concentration of 2.5 mM, ethyl parabens showed a cell survival rate of about 40.5%, and vanillin propionate showed a cell survival rate of about 54.1% higher than this (Fig. 15 ( A) see). Similar results were also shown in the test for dermal fibroblasts (see Fig. 15(B)).
  • Fig. 16 shows the results of L/D fluorescence staining of keratinocytes
  • Fig. 17 shows the results of L/D fluorescence staining of dermal fibroblasts. It can be seen that the results of L/D fluorescence staining in FIGS. 16 and 17 also show the same trend as the results of the WST-1 assay.
  • Example 2 Preparation of a preservative using vanillin alcohol and triacetin
  • Example 1 4-hydroxy-3-methoxybenzyl acetate (vanillin acetate) prepared in 1-2 of Example 1 was synthesized using another substrate. Triacetin was used instead of ethyl acetate as an acyl donor. Like 1-2 in Example 1, the reaction was carried out in an eco-friendly solvent-free system by replacing the solvent by using an excess amount of triacetin, which is a liquid (see FIG. 18: Enzymatic transester using vanillin alcohol and triacetin as reactants. Fire reaction). Through the above reaction, the same vanillin acetate product as the product of Example 1 1-2 may be obtained. Hereinafter, a product containing vanillin acetate obtained by reacting vanillin alcohol and triacetin is expressed as a triacetin product.
  • Triacetin which is used as an acyl donor in the above reaction, is a known GRAS (Generally Recognized As Safe) substance and is widely used as a food additive, and can be used as it is without removing the remaining triacetin after the reaction is completed.
  • GRAS Generally Recognized As Safe
  • glycerol which is produced as a by-product of the above reaction, is a representative cosmetic moisturizing component, so there is no need to remove it after the reaction.
  • Example 2 Compared with the vanillin acetate obtained by the 1-2 method of Example 1, the purity of vanillin acetate is low because triacetin, a substrate added in excess to the material obtained after the reaction, and glycerol, a by-product of the reaction, are mixed together. Nevertheless, the manufacturing method of Example 2 has the advantage that a more economical process can be provided by eliminating the separation process that requires considerable energy and cost.
  • FIG. 19(A) 100 mM vanillin alcohol and triacetin solution, a total volume of 2 mL, an enzyme concentration of 10 U/mL, 40 °C, and the result of the reaction for 5 hours under the conditions of 20 rpm are shown in FIG. 19(A).
  • a triacetin product containing the highest concentration of vanillin acetate a 1 M concentration of vanillin alcohol and triacetin solution, a total volume of 2 mL, an enzyme concentration of 20 U/mL, 70 °C, 400 rpm for 6 hours.
  • the results of the reaction are shown in Fig. 19(B).
  • the triacetin product contains by-products, antibacterial efficacy may be lower than that of the same amount of the preservative obtained by the synthesis method of Example 1 1-2. Accordingly, various triacetin products were synthesized by adjusting the molar ratio of the reactants so that the triacetin product may contain a high concentration of vanillin acetate. The reaction conditions were reacted for 4 hours at 60° C. and 200 rpm in a shaking incubator, and then biosynthesis was terminated. Table 2 shows the molar concentration of vanillin alcohol, the molar ratio of the reactant, the amount of enzyme used in the reaction, the reaction conversion rate, and the purity.
  • Antibacterial efficacy was evaluated in the same manner as in the 2-1 paper disc diffusion assay of Example 1, and the liquid sample was evaluated based on 2 wt%.
  • 3,4-dihydroxybenzyl alcohol was used as an acyl receiver to perform a transesterification reaction.
  • 3,4-dihydroxybenzyl alcohol was used as an acyl acceptor, and ethyl propionate was used as an acyl donor.
  • the reaction was carried out in a solvent-free system by replacing the reaction solvent using an excess of liquid ethyl propionate (see Fig. 20: Enzymatic trans using 3,4-dihydroxybenzyl alcohol and ethyl propionate as reactants. Esterification reaction).
  • 3,4-dihydroxybenzyl propionate was produced as a reaction product.
  • the recovered product 3,4-dihydroxybenzyl propionate (purity 80%) was used to evaluate the antibacterial efficacy.
  • Antibacterial efficacy was evaluated in the same manner as in the 2-1 paper disc diffusion assay of Example 1, and the liquid sample was evaluated based on 2 wt%.
  • 3,4-dihydroxybenzyl propionate showed antibacterial efficacy against all strains of E. coli , S. aureus and C. albicans , showing a broader antibacterial spectrum than vanillin propionate (see Table 4). ).
  • the compounds prepared by the transesterification reaction exhibit lower toxicity than synthetic substances (eg, parabens) used as preservatives, and at the same time, E. coli , S. aureus , C. albicans and A. niger It was confirmed that the antibacterial effect was similar to or better than the existing preservatives against various bacteria such as. Therefore, the compound prepared by the transesterification reaction may be used as a new preservative to replace the existing synthetic material.
  • synthetic substances eg, parabens

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  • Plant Pathology (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Dentistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Environmental Sciences (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Food Preservation Except Freezing, Refrigeration, And Drying (AREA)
  • Cosmetics (AREA)

Abstract

La présente invention concerne un nouvel agent de conservation, et son procédé de préparation. Un agent de conservation selon la présente invention offre une excellente activité antibactérienne contre les bactéries à gram positif, les bactéries à gram négatif, les levures et les moisissures, et est très sûr. De plus, un procédé de production d'un agent de conservation selon la présente invention est un procédé de conversion enzymatique d'un composé phénolique naturel, et présente un taux de conversion de réaction élevé en un court laps de temps avec une petite quantité d'enzyme, même dans des conditions de réaction sans solvant, respectueuses de l'environnement, à température et pression normales.
PCT/KR2020/012784 2019-10-07 2020-09-22 Nouvel agent de conservation, et son procédé de production WO2021071136A2 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
KR10-2019-0124110 2019-10-07
KR20190124110 2019-10-07
KR1020200122337A KR102419005B1 (ko) 2019-10-07 2020-09-22 신규한 보존제 및 이의 제조방법
KR10-2020-0122337 2020-09-22

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WO2021071136A2 true WO2021071136A2 (fr) 2021-04-15
WO2021071136A3 WO2021071136A3 (fr) 2021-06-10

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Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6114377A (en) * 1997-07-17 2000-09-05 E-L Management Corp. Antimicrobial cosmetic compositions
JP3092006B1 (ja) * 1999-04-30 2000-09-25 森永製菓株式会社 新規なカプサイシノイド様物質の合成法
JP2002053408A (ja) * 2000-08-08 2002-02-19 Nissan Chem Ind Ltd カテコール誘導体を含有する工業用抗菌・抗カビ剤、防藻剤及び生物付着防止剤
ES2307420B1 (es) * 2007-04-24 2009-10-20 Universidad De Cadiz Procedimiento de sintesis quimica de capsinoides.
FR2950884B1 (fr) * 2009-10-01 2011-11-11 Oreal Utilisation de derives de vanilline comme conservateur, procede de conservation, composes et composition
WO2012086812A1 (fr) * 2010-12-22 2012-06-28 味の素株式会社 Composés glycosidiques

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