JPH0556324B2 - - Google Patents

Description

[Detailed description of the invention]

(Industrial Application Field) The present invention relates to a multichain surfactant that can form good liposomes and is suitable for biological applications. (Prior Art) Bangham discovered in 1965 that lecithin forms closed vesicles (liposomes) with a diameter of 0.05 to 10 μm consisting of a lipid bilayer membrane in water.
Furthermore, much research has been conducted on liposomes, including their use as a type of ultra-microcapsule material that encapsulates drugs and enzymes, their use as biological membrane models, their use as biologically similar reaction fields, and their application in genetic engineering. Synthetic surfactants such as didodecyldimethylammonium bromide have also been shown to form liposomes in water (T. Kunitake, Y.
Okahata, J.Am.Chem.Soc., 99 , 3860 (1977)). (Problem to be Solved by the Invention) However, liposomes made of phospholipids such as lecithin are sensitive to heat and light and easily decomposed, so when producing, storing, and using liposomes, they may deteriorate and degrade or have poor performance. Moreover, since the lifespan of liposomes is short, quality control such as refrigeration at an appropriate temperature requires time and effort. In addition, liposomes (synthetic bilayer membranes) formed from synthetic surfactants
However, they have many drawbacks such as safety, skin irritation, etc., and cannot be used for applications related to living organisms, and they do not necessarily form a good liposome structure. (Means for solving the problem) The present inventors have conducted extensive research in order to develop a liposome material that forms a fine, stable (long-life) and good liposome structure, and can also be used for biological purposes. As a result, by using a surfactant with a multifunctional structure derived from living organisms, microscopic liposome particles can be obtained, forming a good liposome structure that is stable, biocompatible, and biodegradable. It was discovered that a highly safe derivative can be obtained by using the method, and based on this knowledge, the present invention was completed. That is, the present invention provides the following general formulas [], []
The present invention provides a multi-chain surfactant characterized by comprising at least one kind selected from the compounds represented by [] and [ ].

【formula】

【formula】

[Formula] (In the formula, one of X ¹ and X ² is an amide-bonded alkylamino group, and the remaining one is -
OM (M is an alkali metal, alkaline earth metal, ammonium or organic base, Y ¹ , Y ² and Y ³
One or two of them are amide-bonded alkylamino groups, and the remaining two or one are -OM
(M is an alkali metal, alkaline earth metal, ammonium, or an organic base, one or two of Z ¹ , Z ² and Z ³ are amide-bonded alkylamino groups, and the remaining two or one -OM (M is an alkali metal, alkaline earth metal, ammonium or organic base) In the present invention, the compounds [] to [] above are spicrisporic acid (4,5-dicarboxy-
4-Pentadecanolide; case of []) (Yutaka Ishigami,
Shinsuke Yamazaki, Fragrance Journal) and its lactone ring opened ring acid (3-hydroxy-1,3,4-tetradecanetricarboxylic acid; in the case of []) and agaricic acid (2-hydroxy-1,2,3 -nonadecanetricarboxylic acid (in the case of It can be synthesized by introducing a hydrophilic group structure as a neutralization salt with a metal, ammonium, or an organic base such as morpholine or alkylamine. Normally, according to the culture method, 100% of spicrisporic acid (S-acid) has the chemical structure [ ].
It has a _C10 alkyl group, a lactone ring, and two carboxyl groups, has optical activity, and is insoluble in water. beginning,
In the culture solution, the tribasic acids shown in [] are
Upon heating, one of the carboxyl groups forms an internal ester, forming a dibasic acid with a γ-lactone ring (S-
acid). An open ring acid (O-acid) is obtained by adding an excess aqueous sodium hydroxide solution to the S-acid and heating it, neutralizing it with hydrochloric acid, and extracting the ethyl ether-soluble material. Also,
Agarisic acid (A-acid) contained in the fruit of the basidiomycete Polyporus officinalis is also a tribasic acid with a 100% crystalline structure and a _C18 alkyl group. In the present invention, the group derived from a carboxyl group in the compounds represented by the above general formulas [] to [] has at least one alkyl chain, and the total number of carbon atoms in the alkyl chain is determined from the viewpoint of liposome-forming property. is preferably 4-22, and preferably 8-18.
This alkyl chain may be a straight chain or a branched chain. Further, the compounds represented by the above general formulas [] to [] preferably have at least one group derived from a carboxyl group in the form of a neutralized salt with various bases. The multi-chain surfactant of the present invention further chemically bonds one or two chain alkyl groups to biosoap, and the resulting biosoap derivative acquires great lipophilicity. By neutralizing the remaining carboxyl groups of the derivative with various bases, it is possible to balance the lipophilicity and hydrophilicity of the generated novel multi-chain surfactant molecules and impart liposome-forming ability. Further, more preferred embodiments of the present invention will be described below. The multichain surfactant of the present invention must be oil-soluble in order to form liposomes in water.
a state in which it is emulsified or dispersed in water;
That is, if the surfactant has an HLB of 9 to 13, it will not form a liposome structure but will form micelles in water. However, what is important for liposome formation here is that even though it is oil-soluble, it must also be hydrophilic or water-swellable. In order to form such a chemical structure for forming liposomes, in the case of S-acid and O-acid, a decyl group is originally chemically bonded, so when chemically bonding another chain, a dodecyl group is preferable. It is preferable to neutralize two of the remaining two carboxyl groups with n-hexylamine. Further, if one chain is a decyl group, the remaining two carboxyl groups may be an octylamine salt. On the other hand, when a single chain dodecyl group is introduced into the A-acid, both carboxyl groups of the hydrophilic groups may be converted into sodium salts. Next, when a two-chain alkyl group is introduced to the O-acid, when an n-butyl group is used, the remaining carboxyl group is a butylamine salt. Note that in order to obtain liposome-forming derivatives from these biosoaps, it is desirable to neutralize the carboxyl groups. However, in applications where the solubility and compatibility of multi-chain surfactants with organic solvents and various organic compounds are important, neutralization is not necessarily necessary. In this case, the chain length of the alkyl group for the biosoap may be long or short, and therefore may be a methyl group. (Effects of the Invention) The novel multichain surfactant of the present invention has excellent liposome-forming ability, is biocompatible, biodegradable, and highly safe, and is suitable for biological use. According to this surfactant, it is possible to form a small, stable (long life) and good liposome structure. In addition, the multichain surfactant of the present invention is compatible with a wide range of organic compounds such as various organic solvents, animal and vegetable oils, plant essential oils, and mineral oils, so it can be used in oils and ointments for pharmaceuticals, agricultural chemicals, and cosmetics. Can be used as (Examples) Next, the present invention will be described in more detail based on Examples. Example 1 6.0 g (0.01731 mol) of open ring acid was added to 500
The mixture was placed in a ml eggplant-shaped flask with a slot, 200 ml of dehydrated tetrahydrofuran (THF) was added, and the mixture was stirred with a magnetic stirrer to dissolve. A dropping funnel (with slots) was connected to the flask, and a solution of 2.2 g of equimolar oxalyl chloride and 100 ml of THF was added dropwise from the dropping funnel over 3 hours at room temperature (approximately 25°C). After the dropwise addition was completed, the mixture was heated to 40°C and stirred for about 1 hour. After this, once THF is evaporated and removed,
Wash the residue with a mixture of benzene and hexane, and then rinse again.
THF was added, and a THF solution of 3.21 g (0.01731 mol) of dodecylamine was slowly added dropwise from the dropping funnel at room temperature over 2 hours while stirring. After the dropwise addition was completed, the mixture was heated to 40°C and stirred for about 1 hour. The reaction product was recrystallized from a mixed solvent of 2 parts of ethanol and 1 part of ethyl ether. 4.6 g of white powder was obtained. In the infrared absorption spectrum, absorption of the amide was observed at 1615 and 1555 cm ^-1 , absorption of the carboxyl group split into two at 1720 and 1755 cm ^-1 , and absorption based on the hydrocarbon chain at 2908, 1460 and 1373 cm ^-1 . . The actual value of elemental analysis is C (%) 67.17 (calculated value
67.93), H (%) 11.17 (10.81), N (%) 2.93 (2.7
3)
It was hot. The optical rotation is [α] ²⁸ _D = -16.8 (C = 0.803
g/dl, EtOH). This product was insoluble in water, easily soluble in methanol and chloroform, sparingly soluble in benzene and acetone, and insoluble in hexane. Next, when we prepared a neutralized salt of the remaining carboxyl group in the molecule with an alkali, the sodium salt was highly hydrophilic, the butylamine salt was also water-soluble, but the hexylamine salt was oil-soluble. 1
-(N-Dodecylcarbamoyl)-3-hydroxy-3,4-tetradecanedicarboxylic acid di-n-hexylamine salt is oil-soluble, but it is also slightly hydrophilic, making it difficult to form liposome structures in water. Predicted. Therefore, water was added to the chloroform solution according to a conventional method, the chloroform was evaporated after ultrasonic irradiation, and the mixture was sufficiently stirred using a vortex mixer to obtain a translucent water suspension. The generation of liposomes (synthetic bilayer membranes) was confirmed by electron microscopy (stained with uranyl acetate). Furthermore, closed vesicles with various sizes and shapes ranging from 0.1 to several μm were confirmed. Particle size measurement using a photon correlation method revealed that the average size was 1.5 μm. In addition, differential scanning calorimetry (DSC) shows that there is a gel-liquid crystal phase transition temperature (Tc) and that Tc is
It was found to be 52.1℃. In the above reaction procedure, a two-chain derivative was obtained by esterification using dodecyl alcohol instead of dodecylamine, and liposomes were also formed as a neutralized salt with hexylamine. Example 2 Open ring acid 2.2g in the same manner as Example 1
A THF solution of 2.44 g (0.019 mol) of oxalyl chloride was added dropwise over 4 hours to a THF solution of oxalyl chloride (0.0063 mol), and then 3.56 g (0.019
mol) was added dropwise. The reaction mixture was precipitated by adding hexane, and then purified using an ethanol-hexane system. In the infrared absorption spectrum, in addition to the same absorption of the amide as in Example 1, small absorption of the carboxyl group split into two is seen at 1730 and 1750 cm ^-1 , indicating that all three carboxyl groups in the molecule are absorbed by dodecylamine. It can be seen that it is not amidated. From the nitrogen content determined by elemental analysis, it was estimated that 2.4 of the three carboxyl groups were amidated. After neutralizing the carboxyl group with sodium hydroxide, the monosodium salt was obtained by recrystallization from ether. Formation of liposomes was confirmed using an electron microscope in the same manner as in Example 1. Note that those not considered as neutralized salts are insoluble in water, but are soluble in many organic solvents such as methanol, ethanol, acetone, chloroform, carbon tetrachloride, benzene, hexane, and benzyl alcohol.
It was also confirmed that it is compatible with essential oils such as bergamot oil and lanolin. Therefore, it can be used as an oil agent in cosmetics, and it was also found to have skin softening and fatliquing effects, which are thought to be due to the amide group. Example 3 A benzene solution containing 1.07 g (0.0084 mol) of oxalyl chloride was slowly added dropwise to a benzene solution containing 3.5 g (0.0084 mol) of agaricic acid over 2 hours. After heating at 45°C for 1 hour, remove benzene-insoluble matter by filtration and remove 1.56 g of dodecylamine (0.0084
mol) of benzene solution was slowly added dropwise over 1.5 hours. After the reaction, insoluble matter was purified by concentrating the filtrate and recrystallizing it from methanol. In the infrared absorption spectrum, secondary amide 1720 cm ^-1 at 1615 and 1555 cm ^-1
Absorption of carboxyl groups was observed. It was confirmed by infrared absorption spectrum and elemental analysis that one amide group was introduced into the molecule. By neutralizing the remaining two carboxyl groups of this product with sodium hydroxide, a two-chain surfactant that is sparingly soluble in water was obtained. It was confirmed that liposomes were produced in the same manner as described at the end of Example 1. Example 4 6.0 g (0.0182 mol) of spicrisporic acid was added to 500
The mixture was placed in a ml eggplant-shaped flask, and 200 ml of THF was added to dissolve it. A THF solution containing 6.76 g (0.036 mol) of dodecylamine was slowly dropped into the solution heated to 65° C. from a dropping funnel. After further heating for 2 hours, hexane was added to isolate the reaction product, which was recrystallized from ethanol. In the infrared absorption spectrum, the absorption of the lactone ring disappears, so
It was found that the lactone ring of spicrisporic acid was amidated. Elemental analysis confirmed that two amide groups were introduced into the molecule. Furthermore, an ethanol solution of this product showed no optical rotation, and it was clear that it was racemized by heating.

Claims (1)

[Scope of Claims] 1. A multi-chain surfactant comprising at least one compound selected from the following general formulas [], [] and []. [Formula] [Formula] [Formula] (In the formula, one of X ¹ and X ² is an amide-bonded alkylamino group, and the remaining one is -
OM (M is an alkali metal, alkaline earth metal, ammonium or organic base, Y ¹ , Y ² and Y ³
One or two of them are amide-bonded alkylamino groups, and the remaining two or one are -OM
(M is an alkali metal, alkaline earth metal, ammonium, or an organic base, one or two of Z ¹ , Z ² and Z ³ are amide-bonded alkylamino groups, and the remaining two or one -OM (M is an alkali metal, alkaline earth metal, ammonium or organic base)