KR960015108B1 - Movel sophoro lipid derivatives and the surfactatnt - Google Patents

Abstract

This invention relates to useful sophoro lipid derivatives having the following formula(I) as a surfactant and gelation agent. In the formula, R1 is hydrogen or acethyl group, R2 is fatty acid or lipid group.

Description

[Name of invention]

New vesicle lipid derivatives and surfactants using them

Detailed description of the invention

[Industrial use]

The present invention relates to a lipid derivative and a surfactant using the same as vesicles represented by the following general formula (I), which are useful as water-soluble or oil-soluble surfactants and gelling agents.

(Wherein R ¹ is hydrogen or acetyl group, R ² is hydrogen, aliphatic or aromatic group)

[Prior art]

Conventionally, sorbitan fatty acid esters, sorbitol fatty acid esters, dibenzylidene sorbitol, various alcoholic glycosides, dextrin fatty acid esters and the like are known as surfactants derived from sugars. However, they exist in a large number of isomeric mixtures by side reactions and the like, and since the chemical structure is difficult to specify, it is difficult to reproduce the properties in actual use, and the esterified materials are susceptible to hydrolysis. Natural saccharide surfactants include vesicles, lipids, rhamno lipids, emalzan and the like. These are good surfactants and exhibit inherent properties. Among these, Soropo Lipid is most efficiently produced by culturing Glucose and Triglyceride as substrates by Candida bombicola (more than 70 g per 1 liter of fermentation solution) and is widely studied worldwide.

The vesicle lipids were first reported by P. J. A. Gorins in the culture of yeast Candida (Torulpsis) bombicola [Canadian Journal of Chemistry, 39,846 (1961)]. (Note: Torlobp genus is currently classified as Candida genus).

The vesicle lipids are also relatively in the culture by other yeasts such as Candida bogoriensis, Candida magnolie, Candida gropengisseri, Candida apicola. Large quantities have been reported. [R. Hommel, Biodegradation, 1, 107 (1991).

These vesicle lipids are sugar lipids based on derivatives of sorbose which are always disaccharides in the side chain fatty acid component which is a non-saccharide part. As the fatty acid component, 17-hydroxy octadecanoic acid, 17-hydroxy-9-octadecenoic acid, 13-hydroxycodic acid and the like are known so far. In addition, if the carbon source at the time of culture is appropriately selected, the structure of the side chain portion can be changed.

The vesicle lipid is usually produced by supplying non-aqueous carbon sources such as hydrocarbons and fats and oils, and is mainly produced in the late orientation (normal phase) in which the increase in cell mass is stopped. However, among the yeasts described above, it is also possible to produce lipid vesicles directly from sugars. have. Candida (Torulpsis) bombicola In the cultivation of ATCC 22214, lipids are obtained with about 70 g of parcels per liter of culture by appropriate combination of sugars such as glucose and natural oils such as sunflower oil as a carbon source. Is reported by the Coopers. Applied and Environmntal Microbiology, 47, 173 (1984).

Lipids can also be obtained by using the above-described resting cells of yeast (cells in which growth has stopped). Gobert, et al., Reacted the cells obtained by using a medium made of glucose, yeast extract, urea, etc. with a solution containing several carbon sources, and the lipids of about 18 g of vesicles per liter of the reaction solution. It is reported to be produced. Biotechnology Letters, 6,225 (1984).

It has been reported that the vesicle lipids have an amphiphilic structure, which is a glycolipid, and thus have a great surface activity and strong growth inhibition against bacteria or fungi. R. H Ommel, Applied Microbiology and Biotechnology, 26,199 (1987).

Furthermore, since the sweetener, stepia, has a skeleton as a vesicle, a method of obtaining lipids into vesicles by hydrolysis of stepia has been studied. (Patent Publications 56-39317 and 56-39318). Although vesicle lipid esters are expected to have various industrial uses, they have a drawback of losing their function due to hydrolysis when used in a neutral to alkaline region and at high temperatures.

[Problem to Solve Invention]

The inventors of the present invention provide a lipid derivative as a raw material, which reacts with various amines, and has high chemical stability and facilitates intermolecular hydrogen bonding. The task is to provide.

[Means for solving the problem]

MEANS TO SOLVE THE PROBLEM The present inventors came to complete this invention as a result of repeating research in order to solve the said subject. That is, in the present invention, a lipid derivative is provided as a vesicle of the following general formula (I).

(Wherein R ¹ represents hydrogen or an acetyl group, R ² represents hydrogen, a fatty acid group or an aromatic group) In the present invention, the above-mentioned vesicles are provided with a surfactant derived from a lipid derivative.

The vesicle lipid derivative according to the present invention is prepared by using lipid as vesicles represented by the following formula (1) or (2) obtained by a conventionally known culture method.

In the preparation of the compound of the present invention using lipid as a raw material of the vesicle of the formula (1), the preparation of the compound of the present invention using N-methyl-2-chloro-pyridinium urethotite in a methylene chloride solvent was used as the raw material vesicle. And by reacting with an amine compound in the presence of tributylamine. In this case, the reaction formula is represented by the following equation.

In said formula, R <^2> is hydrogen, an aliphatic group, or an aromatic group. As an aliphatic group, an alkyl group, an alkenyl group, and a cycloalkyl group are mentioned, The carbon number is 1-18 normally. Examples of the aromatic group include alkyl groups such as phenyl, tril and naphthyl, and alkyl groups such as benzyl and phenethyl.

The vesicle obtained in the above reaction is a compound in which R ¹ is represented by an acetyl group in the general formula (I) of the lipid derivative. In preparing a compound of the present invention using the lipid derivative as a raw material of the lactone vesicles of Formula (2), first, the lipids are hydrolyzed into lactone vesicles of the general formula (2) and converted into lipids into acidic vesicles. Lipids of the acid type vesicles obtained by this hydrolysis are represented by the following formula.

The lipid derivatives (R ¹ = H in formula (I)) can be obtained by reacting the lipids with R ² NH _{2 in the} same manner as the above.

Next, an Example is given and this invention is demonstrated in detail.

[Comparative Example]

[Recipes manufacturing method by parcel]

100 g of glucose, 100 g of corn oil, 3 g of salt lake ammonium phosphate, 1 g of dihydrogen phosphate, 5 g of magnesium sulfate, and 5 g of yeast extract were added to a total amount of 1 L, and put into a 2 L fermentation tank to sterilize it. The fermentation broth was inoculated with a cell solution (50 mL) of Candida bombicola ATCC 22214 obtained by incubating the culture at 30 ° C. for 48 hours in a culture medium having the same composition. The temperature was 30 ° C., agitation water 500 rpm, and aeration volume 1 L /. Fermentation was started under min conditions. Fermentation was stopped 7 days after the start of the culture.

The culture solution taken out of the fermentor was centrifuged, the culture drug and the cells were separated, and the culture solution was mixed with a double volume of ethyl acetate and a separatory funnel, and lipids were extracted with vesicles. The separated upper layer was separated, the solvent was removed, and lipids were obtained with about 26 g of the syrup-like vesicles remaining. Hexane was added to the lipids with this vesicles, followed by repeated washing and dissolving and removing a substantial amount of raw material oil and the like mixed as impurities.

Lipids were subjected to hexane treatment, and the lipids were subjected to silica gel column chromatography to develop and elute in a chloroform-acetone system (multiple stages of acetone concentration).

Liposomal lipids were not eluted with chloroform alone, but were detected in chloroform-acetone mixed solution. The elution behavior and the degree of purification were determined by thin layer chromatography using chloroform-methanol-water as the developing solvent. At this time, the lipid component was confirmed by α-naphthol reagent with various vesicles on a thin plate.

After the purification operation, the recovered parcels had a lipid of about 18 g. Confirmation of lipids with the obtained vesicles was basically carried out for comparing Rf values in thin layer chromatography with known vesicles. The content of acid form [compound of formula (1)] at the time of lipid vesicle obtained by pH titration was 44%, and the lactone type [compound of formula (2)] was 44%.

A 0.1% aqueous solution (pH 6.8) was prepared, and the surface tension was measured by the Will Helm method (Shimazu ST-1 type). As a result, 37.0 mN / m (25 ° C) and spin drop type interfacial tension meter (Core Laboratories 500 type) The interfacial tension with respect to hexadecane at) was measured to be 5.1 mN / m (25 ° C).

Example 1

As a vesicle obtained in Comparative Example, 1.3 g of lipid (acid type) was placed in a two-necked 50 ml flask, and 20 ml of methylene chloride was added to dissolve, followed by 0.23 g of benzylamine, 0.43 g of tributylamine, and N-methyl-2-chloro-. After adding 0.5 g of pyridinium yodozit, a drying tube containing reflux cooler and pellets of kallium hydroxide was attached to the flask, and the flask was kept at 40 ° C. using a constant temperature water bath and stirred for 5 hours while stirring with a magnetic stirrer. I was.

After completion of the reaction, saturated aqueous sodium bicarbonate solution was added to the reaction mixture to neutralize, and further washed with water until the pH became neutral. After the water was removed, anhydrous sodium sulfate was added, dried, filtered, methylene chloride was removed by evaporation, and dried to obtain 1.4 g of lipid benzylamide as a vesicle.

TLC analysis of the preparation using Merck Co., Ltd. Kigel gel 60 (thickness 0.5 mm) and a mixed solvent of CHCL ₃ : CH ₃ OH: H ₂ O = 65: 15: 2 as the developing solvent. The six spots were separated by the process. Among the two large peaks, the spot of Rf0.7 below was lipid benzylamide in vesicles, confirmed by IR analysis from the presence of 1660 and 1570 cm ⁻¹ of amide absorption. Furthermore, separation was carried out in HPLC.

In other words, Waters Corporation (Waters Co.) We share (Bondashere) 5μNH ₂ -100A writing a column of acetonitrile as a developing solvent by HPLC system (system): about once in 1 (flow rate 10㎖ / min): water = 9 16 mg of lipid benzylamide was obtained from the 50 mg reaction mixture with vesicles. The surface tension (25 占 폚) of the 0.1% aqueous solution was 39.0 mN / m, and the interfacial tension (25 占 폚) for hexadecane was 2.5 mN / m.

Example 2

1.3 g of the lipid preparation was taken into a two-neck 50 ml flask by dissolution of the vesicle obtained from the comparative example, and 20 ml of methylene chloride was added thereto to dissolve 0.34 g, 0.43 g of tributylamine and N-methyl-2-chloro-pyridinium iodozide. After adding 0.5 g, a drying tube containing a reflux condenser and potassium hydroxide pellets was attached to the flask, and the flask was kept at 40 ° C. using a constant temperature water bath and reacted for 5 hours while stirring with a magnetic stirrer.

After completion of the reaction, saturated aqueous sodium bicarbonate solution was added to the reaction mixture to neutralize, and further washed with water until the pH became neutral. After removing water, anhydrous sodium sulfate was added, dried, filtered, methylene chloride was evaporated and dried to obtain 1.5 g of a preparation of lipid decylamide as a vesicle.

TLC analysis of the preparation was carried out using a Merck Kiesel gel 60 (5 mm thick), and a mixed solvent of CHCl ₃ : CH ₃ OH: H ₂ O = 65: 15: 2 as a developing solvent. Six spots were separated.

The spot of Rf 0.85 in the two large peaks was identified from the presence of 1654 and 1555 cm ⁻¹ amide uptake by IR analysis, which is lipid decylamide into vesicles. Furthermore, two separations were performed in HPLC.

That is, 21 mg of lipid decylamide was obtained as vesicles from about 50 mg of the reaction mixture under the same conditions as in Example 1 by Waters HPLC. Even when a 0.1% aqueous solution was prepared, it usually became turbid, but no precipitation occurred. The surface tension (25 degreeC) of the 0.1% solution was 33.1 mN / m, and the interface tension (25 degreeC) with respect to hexadecane was 3.3 mN / m.

Example 3

1.0 g of lipid was added to a 100 ml round flask with lactone vesicles obtained in the comparative example, and 10 ml of dry methanol was added to dissolve. Separately, 10 ml of a solution of 140 mg of metal natrium dissolved in dry methanol was slowly added, followed by Pellet-dried tube was attached to the flask and allowed to react for 8 hours while stirring with a magnetic stirrer at room temperature. A cation exchange resin (Aorgano Co., Ltd. Ambarist) was added to the reaction mixture to neutralize, fractionated and evaporated to dryness.

It was eluted by silica gel column chromatography on a chloroform-acetone system. In this way, the ring opening was ring-opened, the acetyl group was hydrolyzed to obtain lipid [compound of formula (4)], which was changed to a free hydroxyl group, and was treated in the same manner as in Example 2 to prepare lipid decylamide as a vesicle 1.3 g was obtained.

A 0.1% aqueous solution was prepared, and the surface tension (25 ° C.) and the interfacial tension with respect to hexadecane were measured. The results were 33.1 mN / m and 3.3 mN / m, respectively.

[Effects of the Invention]

The new vesicle lipid derivatives according to the present invention have superior interfacial tension lowering ability as compared to existing vesicle lipid lipids, and can be used as water-soluble or oil-soluble surfactants and gelling agents.

Claims (2)

Lipid derivatives as vesicles represented by the following general formula (I) (Wherein R ¹ is hydrogen or acetyl group, R ² is hydrogen, aliphatic acid or aliphatic) Surfactant consisting of lipid derivatives with vesicles of the general formula (I) (Wherein R ¹ is hydrogen or acetyl group, R ² is hydrogen, aliphatic group or aliphatic)