KR101861785B1 - manufacturing method of isoserinol amide derivatives - Google Patents

Abstract

The present invention relates to a method for preparing an isoserinol amide derivative represented by chemical formula 1, wherein the method comprises the steps of: A) mixing ethyl ester represented by chemical formula 2 with 3-amino-1, 2-propanediol; B) adding 0.1 to 15% of a novozyme 435 (Novo) enzyme on the basis of weight with respect to an amount of the ethyl ester represented by the chemical formula 2 to be reacted; C) removing the enzyme after completion of the reaction; D) adding an organic solvent to perform heating and agitation; and E) performing filtering. A novel isoserinol fatty acid amide derivative having an antibacterial or anti-inflammatory activity and an excellent skin moisturizing effect can be provided.

Description

Manufacturing methods of isoserinol amide derivatives < RTI ID = 0.0 >

The present invention relates to a novel process for preparing a fatty acid amide compound of isoserinol and a cosmetic composition comprising the compound.

Generally, monoglycerides of fatty acids have been used as emulsifiers in foods and cosmetics, and some of them may exhibit antibacterial activity depending on the kinds of fatty acids. Fatty acid glycerides and related fatty acid derivatives are also known to interact with endocannabinoid 2-arachidonoylglycerol (2-AG) in pain sensitivity and anti-inflammatory regulation (JM Walker et al. , Chemistry and Physics of Lipids 121 (2002) 159-172).

On the other hand, vegetable oils are widely used as raw materials for cosmetics, which refers to oil obtained from the fruit or seed of a plant by a compression method and a distillation method. It contains a large amount of various unsaturated fatty acids, which enhances the skin's amenity, strengthens the barrier of skin, moisturizes skin, and supplies skin nutrition. In addition, free fatty acids contained in vegetable oils, especially palmitic acid, lauric acid, ricinoleic acid, and linolenic acid have a strong anti-inflammatory effect It is known.

Although isoserinol is widely used in cosmetics by mixing with the vegetable oil, research on cosmetics using fatty acid derivatives of isoserinol has not been conducted.

Accordingly, the present inventors have produced a fatty acid amide derivative of isoserinol and a pharmaceutical and cosmetic composition having excellent antimicrobial, anti-inflammatory and skin moisturizing effects. In addition, the fatty acid amide derivative of the isoserinol is expected to have various potential effects depending on the type of fatty acid, and it is confirmed that the fatty acid amide derivative exhibits more stability in the formulation due to the amide bond.

KR 1020170039372 A US 9333230 B2

The present invention relates to novel isoserinol fatty acid amide derivatives having antibacterial or antiinflammatory activity and excellent skin moisturizing effect, a process for producing the same, a composition capable of being used as an active ingredient or an emulsifier in medicines, cosmetics, foods and the like by providing an emulsifier composition containing the same And to provide the above objects.

The present invention relates to a method for producing an isoserinolamide derivative,

A) mixing an ethyl ester represented by the following formula (2) with 3-amino-1,2-propanediol;

B) Novobyme 435 (Novozyme 435, Novo);

C) removing the enzyme after completion of the reaction;

D) heating with an organic solvent and stirring;

E) filtering the mixture to obtain an isoserinolamide derivative represented by the following general formula (1).

≪ Formula 1 >

(2)

(In the above formulas (1) and (2), R is a straight chain alkyl group or a straight chain alkenyl group of C ₅ to C ₂₉ .

In the present invention, the compound represented by the general formula (1) is preferably N- (2,3-dihydroxypropyl) decanamide, N- (2,3-dihydroxypropyl) 2,3-dihydroxypropyl) dodecanamide, N- (2,3-dihydroxypropyl) tetradecanamide, N- (2,3-dihydroxypropyl) dodecanamide, N- 2,3-dihydroxypropyl) palmitamide, N- (2,3-dihydroxypropyl) stearamide, N- (2,3-dihydroxypropyl) stearamide, N- (2,3-dihydroxypropyl) oleamide).

The present invention also provides a cosmetic composition comprising the isoserinolamide derivative prepared by the above-described method as an emulsifier. The cosmetic composition may be provided as a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing oil, powder foundation, emulsion foundation, wax foundation and spray.

The isoserinolamide derivative according to the present invention is a fatty acid amide compound having not only an action of an emulsifying stabilizer but also a anti-inflammatory or antimicrobial function, and a pharmaceutical composition and a cosmetic composition containing the same have excellent antibacterial, anti-inflammatory and skin moisturizing effects.

Hereinafter, the isoserinolamide derivative according to the present invention and a method for producing the same will be described in detail.

The present invention provides a compound represented by the following formula (1).

     ≪ Formula 1 >

Wherein R is a straight chain alkyl group or a straight chain alkenyl group of C ₅ to C ₂₉ .

The present invention relates to a process for the preparation of a compound of formula (I), which comprises: mixing an ethyl ester represented by the following formula (2) and 3-amino-1,2-propanediol; Adding an enzyme (Novozyme 435, Novo) and then reacting; Removing the enzyme after completion of the reaction; Adding organic solvent and heating and stirring; Wherein the isoserinolamide derivative is represented by the following formula (1).

(2)

The present invention also provides a cosmetic composition comprising the isoserinolamide derivative represented by Formula 1 as an emulsifier.

The compound represented by the formula (1) has an anti-inflammatory and antibacterial function as well as an emulsion stabilizer.

In the present invention, an enzyme may be added to produce the compound of Formula 1, followed by an enzyme reaction. As the enzyme, Lipase B (trade name: Novozym435) produced by culturing Aspergilus niger with a lipase gene of Candida antarctica and immobilized on an acrylic resin may be used, and 0.1 to 15 %, Preferably 0.3 to 10%, more preferably 0.5 to 5%. The enzyme is added and reacted at 25 to 75 ° C, preferably 35 to 65 ° C, for 12 to 48 hours, more preferably 24 to 36 hours.

The present invention provides a cosmetic composition comprising the compound represented by Formula 1 as an emulsifier and a method for producing the emulsifier.

The cosmetic composition of the present invention contains components conventionally used in cosmetic compositions and is usually used in cosmetics such as antioxidants, bactericides, dyes, preservatives, stabilizers, thickeners, moisturizers, solubilizers, inorganic salts, vitamins, Phosphorous additive, and carrier. There is no limitation on the route of administration, but local administration such as transdermal administration or subcutaneous administration for whitening purposes is preferable.

The cosmetic composition may be prepared in any form conventionally prepared, for example, such as a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, a surfactant- A powder foundation, an emulsion foundation, a wax foundation, and a spray, but the present invention is not limited thereto. More specifically, it can be manufactured in the form of softening agent, convergent lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder.

The cosmetically effective carrier contained in the cosmetic composition of the present invention may be a commonly used carrier depending on the formulation.

When the formulation of the present invention is an ointment, paste, cream or gel, an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, Lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as the carrier component when the formulation of the present invention is a powder or a spray, Propane / butane, or dimethyl ether. ≪ RTI ID = 0.0 >

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, and in particular fatty acid esters of cottonseed oil, peanut oil, corn oil, olive oil, castor oil and sesame oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.

In the case where the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

Hereinafter, the present invention will be described in more detail with reference to examples. However, the following examples are intended to illustrate the present invention, and the scope of the present invention is not limited thereto.

< Example  1 > to < Example  6> fatty acid chloride and 3-Amino-1,2- propanediol  Method for the chemical production of amide compounds

10.1 g of 3-amino-1,2-propanediol (MW = 91.11) is added to a glass reactor, and 50 ml of dichloromethane is added thereto. After dissolution, 10.2 g of triethylamine (MW = 101.19) is added. The temperature of the reaction solution was adjusted to 15 ° C or lower, the fatty acid chloride as the reaction compound of each of Examples 1 to 6 in the following Table 1 was slowly added dropwise, and the reaction solution was stirred at 35 ° C for 4 hours.

After completion of the reaction, the reaction mixture is adjusted to a pH of 2 to 3 using a 10% HCl solution and then washed with purified water and 5% sodium bicarbonate solution. Afterwards, use purified water and wash until neutrality. The organic layer was dehydrated with sodium sulfate anhydrous and vacuum distilled to prepare the target compound.

Example Fatty acid chloride Input (g) Yield (g) yield(%) Appearance Compound name  One Decanoyl chloride 19.22 20.40 82.5 Light yellow wax N- (2,3-dihydroxypropyl) decanamide 2 Lauroyl chloride 22.04 23.09 83.8 Light white solid N- (2,3-dihydroxypropyl) dodecanamide 3 Myristoyl chloride 24.87 25.55 84.1 Light white solid N- (2,3-dihydroxypropyl) tetradecanamide 4 Palmitoyl chloride 27.70 27.69 83.4 Light white solid N- (2,3-dihydroxypropyl) palmitamide 5 Stearoyl chloride 30.53 30.45 84.5 Light white solid N- (2,3-dihydroxypropyl) stearamide 6 Oleoyl chloride 30.32 30.49 85.1 Light yellow wax N- (2,3-dihydroxypropyl) oleamide

< Example  7 > to < Example  12> Preparation of fatty acid ethyl ester and 3-Amino-1,2-propanediol by enzymatic reaction

10.1 g (MW = 91.11) of each of the fatty acid ethyl esters of the following Reaction Compounds 7 to 12 and 3-Amino-1,2-Propanediol are placed in a glass reactor and 20 ml of Tert-Butanol is added as a solvent. Then, 0.202 g of Novozyme 435 enzyme (Novozyme 435, Novo) was added thereto, followed by shaking at 45 ° C for 24 hours. After confirming that the spot of the fatty acid ethyl ester completely disappeared after the reaction, the enzyme of the reaction mixture was filtered with 100 mesh sieve, and then 500 ml of acetonitrile was added. After stirring for 30 minutes, the reaction mixture was allowed to stand at 10 ° C or lower for 4 hours . After filtration of the precipitate, acetonitrile was removed to obtain an enzyme reaction product of fatty acid ethyl ester and 3-amino-1,2-propanediol.

Example Cholesterol in fatty acids Input (g) Yield (g) yield(%) Appearance 7 Decanoic acid ethyl ester 33.31 20.28 74.6 Light yellow wax 8 Lauric acid ethyl ester 37.97 23.42 77.3 Light white solid 9 Myristic acid ethyl ester 42.64 25.40 76.0 Light white solid 10 Palmitic acid ethyl ester 47.30 30.60 83.8 Light white solid 11 Stearic acid ethyl ester 51.97 32.09 80.9 Light white solid 12 Oleic acid ethyl ester 51.63 31.89 80.9 Light yellow wax

The NMR and mass data of the compounds obtained by the above Examples 1 to 6 and the data of the compounds obtained by the methods according to Examples 7 to 12 were confirmed to be the same compounds.

< Example  13> to < Example  18> Vegetable oil and  3-Amino-1,2- propanediol  Production method using enzyme reaction

50 g of each vegetable oil and the amount of 3-Amino-1,2-Propanediol (MW = 91.11) as determined in Examples 13 to 20 were placed in a glass reactor and 100 ml of Tert-Butanol was added as a solvent. Add 4 g of Novozyme 435 enzyme (Novozyme 435, Novo) and incubate at 45 ° C with shaking for 24 hours. After the reaction, confirm that the spot of the oil part disappears completely through the TLC confirmation test. The enzyme of the reaction solution is filtered with a 100 mesh sieve, 1,000 ml of purified water is added, and the mixture is stirred for 30 minutes and then left at 10 ° C or lower for 4 hours.

The aqueous layer was separated and washed twice. The residue was distilled off under a vacuum of 60 ° C to obtain a complex fatty acid amide compound, which is an enzyme reaction product of 3-Amino-1,2-propanediol in vegetable oil.

Example Oil Input (g) Yield (g) yield(%) Appearance 13 Camellia japonica seed oil 161.58 169.58 87.1 Mythical liquid wax 14 Cocos nucifera (coconut) oil 120.82 133.15 86.5 Light yellow wax 15 Ricinus communis (castor) seed oil 170.49 179.16 88.0 Light yellow wax 16 Canola oil 161.42 165.35 85.0 Light yellow wax 17 Olea europaea (olive) fruit oill 159.56 169.58 87.1 Light yellow wax 18 Elaeis guineensis (palm) oil 155.05 133.15 86.5 Light yellow wax

Test Example  1 to 3, Comparative Example  One. emulsion  Produce

An oil-in-water type emulsion was prepared according to a conventional method with the composition shown in Table 4 below.

ingredient Content (% by weight) Test Example 1 Test Example 2 Test Example 3 Comparative Example 1 N- (2,3-dihydroxypropyl) decanamide 1.0 - - - N- (2,3-dihydroxypropyl) tetradecanamide - 1.0 - - N- (2,3-dihydroxypropyl) stearamide - - 1.0 - 3-Amino-1,2-propanediol - - - 1.0 Cetearyl alcohol 3.0 Stearic acid 2.0 Glyceryl monostearate 1.5 Isopropyl palmitate 3.0 Squalane 3.0 Dimethicone 0.5 Cyclomethicone 1.5 Polyoxyethylene sorbitan monostearate 0.5 Hydrogenated castor oil 0.2 Carboxyvinyl polymer 0.2 Triethanolamine 0.1 1,2-hexanediol 0.2 1,3-propanediol 5.0 Butylene glycol 5.0 Pigment Suitable amount Spices Suitable amount Purified water Balance

Experimental Example  1. Antibacterial test

1) Paper disc antibacterial test against acne bacterium

Amino-1,2-propanediol was diluted with 70% ethanol to concentrations (0.25%, 0.5%, 1%) of the above Examples 1, 3, 5 and 3-Amino-1,2-propanediol. 10 ml of an acne bacterium, Propionibacterium Acnes (P. acnes), was inoculated into liquid medium (GAM Broth, Gifu Anaerobic Medium Broth) and subcultured three times at 37 ° C for 24 hours to obtain an antimicrobial activity test strain Respectively. A paper disc was dropped on the liquid medium in which the strain was cultured, and 20 μl of the test solution was added dropwise to the paper disc. Thereafter, the cells were cultured at 37 ° C for 72 hours, and the clear zone diameter of the bacteria produced around the discs was measured to compare antimicrobial activities. The results are shown in Table 5 below. At this time, phenoxy ethanol was used as a positive control to compare and contrast the antibacterial activities of the mixtures.

Growth inhibition ring size (cm) 0.25% 0.5% One% Example 1 0.8 1.1 1.8 Example 3 1.0 1.4 2.0 Example 5 0.9 1.2 1.9 3-Amino-1,2-propanediol 0.6 1.0 1.7 Phenoxyethanol 0.7 1.1 1.6

From the results shown in Table 5, it was confirmed that Examples 1, 3, and 5 exhibited excellent antimicrobial activity against acne P. acenes as compared to the positive control and 3-Amino-1,2-propanediol.

2) Minimum inhibitory concentration measurement for acne bacteria

In order to further demonstrate the antimicrobial activity of the above Examples 1, 3, 5 and 3-Amino-1,2-propanediol against acne bacteria, a minimal inhibitory concentration (MIC) test for acne bacteria was conducted .

Escherichia coli (E. Coli, ATCC 8739) and Pseudomonas aeruginosa (AT. Pneumoniae, ATCC 9027) were used for the strains Staphylococcus aureus and Staphylococcus epidermidis. Each strain was prepared by sterilizing a nutrient broth solution, and each cultured strain was inoculated into 10 ml of a nutrient broth solution, and cultured in a shaking incubator for 24 hours to prepare a strain for antibacterial experiment Respectively. Example 1, 3, 5 and 3-Amino-1,2-propanediol were added to the medium to a concentration of 0.01 to 1% by weight, and then sterilized to obtain S. aureus, S. cerevisiae, and P. aeruginosa at a concentration of about 1 × 10 ⁸ CFU / ml. epidermidis, E. coli and Ps. aeruginosa was inoculated and cultured under aerobic conditions at 37 ℃ for 24 hours. As a control group, no experimental solution was added. In order to confirm the growth of each strain in the cultured media, approximately 1 ml of each sample was placed in a sterile room and dispensed into a sterilized cuvette. Absorbance was measured at 650 nm using a spectrophotometer, And the minimum inhibitory concentration of each strain was measured and compared with that of the control. The results for the minimum inhibitory concentration (% by weight) measurement are shown in Table 6 below.

Test strain Minimum inhibitory concentration (% by weight) Example 1 Example 3 Example 5 3-Amino-1,2-propanediol Control group S. aureus 0.20 0.07 0.18 0.32 0.91 S. epidermidis 0.16 0.08 0.12 0.21 0.87 E. coli 0.30 0.21 0.25 0.50 1.34 Ps. aeruginosa 0.18 0.14 0.16 0.24 1.10

From the results shown in Table 6, it was confirmed that Examples 1, 3, and 5 exhibited excellent antibacterial activity at a concentration of 0.3% by weight or less with respect to acne bacteria, and showed antibacterial activity as compared with the control group and 3-Amino-1,2-propanediol It is confirmed that it is excellent.

Experimental Example  2. Anti-inflammatory test

In order to confirm the anti-inflammatory effect of Example 3, Nitric oxide (NO) formation inhibition test was performed by GRIESS using mouse macrophage RAW264.7 cell line (ATCC number: CRL-2278). RAW264.7 cells were subcultured several times, placed in a 24-well plate at a concentration of 3 × 10 ⁵ cells / well, and cultured for 24 hours. Thereafter, the cell culture medium of Example 3 was diluted to a final concentration of 1, 10 and 100 ppm. At this time, L-NMMA (L-NG-Monomethylarginine), which is an inhibitor of NO production, was treated together as a positive control and cultured for 30 minutes. Lipopolysaccharide (LPS) 100 μl of the supernatant was transferred to a 96-well plate, and 100 μl of GRIESS solution was added thereto at room temperature for 10 minutes. The absorbance at 540 nm was measured, and the results are shown in Table 7 below.

division NO production inhibition rate (%) Example 3 1 ppm 12.04 10 ppm 20.14 100 ppm 27.57 L-NMMA (10 ppm) 40.04 The control (10 ppm) -

From the results shown in Table 7, it can be seen that the activity of Example 3 is lower than that of L-NMMA, which is a typical anti-inflammatory drug, but exhibits excellent activity. As the concentration increases, Respectively.

Experimental Example  3. Evaluation of skin moisturizing efficacy

To evaluate the skin moisturizing effect, 20 adult men and women aged 30 to 50 years old were divided into four sections of 2 × 2 cm ^{2 on} both sides of the upper and lower portions, and the cosmetic compositions of Test Examples 1 to 3 and Comparative Example 1 were applied A corneometer (Courage + Khazaka electronic GmbH), a moisture analyzer, was used to measure the amount of water present in the skin at constant temperature and humidity (24 ° C, 40% relative humidity) at 120 minutes and 240 minutes after application The skin moisture was measured 5 times each time, and the average value was obtained. The results are shown in Table 8 below. The increment of the measured value was expressed as a percentage (%) when a certain time elapsed after coating, based on the coneometer value measured just before the start of the test.

division Percutaneous water increase rate (%) 120 minutes after application 240 minutes after application Test Example 1 26.4 24.5 Test Example 2 27.3 25.7 Test Example 3 26.0 23.9 Comparative Example 1 24.5 23.4

As shown in Table 8, Test Examples 1 to 3 had a higher rate of increase in transdermal water than Comparative Example 1, and thus, it was confirmed that there was an excellent moisturizing improvement effect.

Experimental Example  4. Evaluation of emulsion stability

The long-term storage stability for Test Examples 1 to 3 was evaluated in a constant temperature bath (-5 DEG C, 37 DEG C, cycle) and room temperature, and the results are shown in Table 9 below. The cycle evaluation allowed circulation of 45 ° C at intervals of 24 hours at -5 ° C.

Stability -5 ℃ Room temperature 37 ℃ cycle 1 week 4 weeks 1 week 4 weeks 1 week 4 weeks 1 week 4 weeks Test Example 1 Good Good Good Good Good Good Good Good Test Example 2 Good Good Good Good Good Good Good Good Test Example 3 Good Good Good Good Good Good Good Good

As clearly confirmed from the stability test result table in Table 9, it was confirmed that Test Examples 1 to 3 were very stable in emulsion stability.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments. Accordingly, the embodiments disclosed herein are intended to be illustrative rather than limiting, and the spirit and scope of the present invention are not limited by these embodiments. The scope of protection of the present invention should be construed according to the following claims, and all the techniques within the scope of equivalents should be construed as falling within the scope of the present invention.

Claims (8)

In the method for producing the isoserinolamide derivative,
A) mixing an ethyl ester represented by the following formula (2) with 3-amino-1,2-propanediol;
B) adding Novozyme 435 (Novo) 435 (Novozyme 435, Novo) enzyme in an amount of 0.1 to 15% based on the weight of the ester of erucic acid represented by the formula 2, and then reacting;
C) removing the enzyme after completion of the reaction;
D) heating with an organic solvent and stirring;
E) filtration to obtain an isoserinolamide derivative represented by the following general formula
&Lt; Formula 1 >

(2)

(In the above formulas (1) and (2), R is a straight chain alkyl group or a straight chain alkenyl group of C ₅ to C ₂₉ .
delete The method according to claim 1, wherein the enzyme is added in step B), and the mixture is reacted with stirring at 35 to 65 ° C
The method according to claim 1, wherein the enzyme is added in step (B) and the mixture is reacted for 12 to 48 hours with stirring
The method according to claim 1, wherein in step (D), the organic solvent is acetonitrile
The method according to claim 1,
The compound represented by the formula (1)
N- (2,3-dihydroxypropyl) decanamide, N- (2,3-dihydroxypropyl)
N- (2,3-dihydroxypropyl) dodecanamide, N- (2,3-dihydroxypropyl) dodecanamide,
N- (2,3-dihydroxypropyl) tetradecanamide, N- (2,3-dihydroxypropyl) tetradecanamide,
N- (2,3-dihydroxypropyl) palmitamide, N- (2,3-dihydroxypropyl)
N- (2,3-dihydroxypropyl) stearamide (N- (2,3-dihydroxypropyl) stearamide) or
Characterized in that it is N- (2,3-dihydroxypropyl) oleamide.
A cosmetic composition comprising an isoserinolamide derivative prepared according to any one of claims 1 and 6 as an emulsifier
The cosmetic composition of claim 7, wherein the cosmetic composition is selected from the group consisting of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing oils, powder foundations, emulsion foundations, wax foundations and sprays At least one cosmetic composition selected from