JPH0940632A - Diacylglycerol derivative - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject new compound having polyvalent hydroxyl group, exhibiting excellent storage stability and useful as a vesiculation agent such as liposome or fat emulsion and as a raw material for pharmaceuticals, agrochemicals and cosmetics. SOLUTION: The objective diacylglycerol derivative is expressed by formula I [X is group of formula II (M is an univalent counter ion) of formula III (R<3> is H or methyl); R<1> C(=O) and R<2> C(=O) are each an acyl residue of a 3-30C fatty acid; (n) is 2-6], e.g. the compound of formula IV. The compound of formula I wherein X is a group of formula II can be produced e.g. by reacting a diacylphosphatidyl ethanolamine having R<1> (=O) and R<2> C(=O) groups such as dimyristoylphosphatidyl ethanolamine with a sugar lactone such as D-gulonic acid-γ-lactone.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a diacylglycerol derivative having a polyvalent hydroxyl group. The diacylglycerol derivative of the present invention is used as an endoplasmic reticulum forming agent such as a liposome or a fat emulsion, and as a raw material for medicines, agricultural chemicals, cosmetics and the like.

[0002]

2. Description of the Related Art Liposomes are phospholipid bilayer vesicles, and their applications have been tried in various fields. In particular, attention is focused on its application to drug carriers, diagnostic / detection carriers, and the like, and improvement of liposome stability and maintenance of blood concentration have become major issues. By the way, diacylglycerol derivatives are widely used as a membrane-forming component of liposomes, but diacylglycerol derivatives having a polyvalent hydroxyl group represented by the general formula (1) described later are not known.

[0003]

An object of the present invention is to provide a new and useful diacylglycerol derivative having a polyvalent hydroxyl group.

[0004]

DISCLOSURE OF THE INVENTION The present invention is a diacylglycerol derivative represented by the following general formula (1).

Embedded image (In the formula (2), M represents a monovalent counter ion, and in the formula (3), R ³ represents a hydrogen atom or a methyl group). R ¹ C (═O) and R ² C (═O) represent an acyl residue of a fatty acid having 3 to 30 carbon atoms and may be the same or different. n represents a positive number of 2 to 6. )

The acyl residue of the fatty acid represented by R ¹ C (═O) or R ² C (═O) in the general formula (1) is an acyl residue having 3 to 30 carbon atoms, preferably 8 to 20 carbon atoms. is there.
Specific examples of such acyl residues include propionic acid, butyric acid, caproic acid, caprylic acid, pelargonic acid, capric acid, undecanoic acid, lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, behenic acid. Acyl residues of saturated fatty acids such as acids, lignoceric acid, solotic acid, montanic acid, melissic acid, 2-ethylhexanoic acid; oleic acid, linoleic acid, linolenic acid, erucic acid, 2,4-octadecadienoic acid, etc. An acyl residue of an unsaturated fatty acid; an acyl residue of a branched fatty acid such as isostearic acid; an acyl residue of a fatty acid having a hydroxyl group in the alkyl group such as ricinoleic acid and 1,2-hydroxystearic acid. R ¹ C (═O) and R ² C (═O) may be the same or different.

When the diacylglycerol derivative of the present invention is used as a membrane-forming component of liposome, R ¹ C
The acyl residue of the fatty acid represented by (= O) or R ² C (= O) is capable of forming stable liposomes, and therefore myristic acid, palmitic acid, stearic acid, oleic acid, 2,4-octadeca An acyl residue such as dienoic acid is preferable, and an acyl residue such as palmitic acid, stearic acid and oleic acid is particularly preferable.

In the general formula (2), M is a monovalent counter ion and is not limited as long as it is a monovalent cation which is a counter ion of PO ₄ ⁻ . Specifically,
H ⁺ ; alkali metal ions such as Na ⁺ and K ⁺ ; NH ₄ ⁺ , (C
^{_{H 3) 4 N +, (}} C 2 H 5) 4 N +, (C 3 H 7) 4 N +, (CH 3) 3 N +
_{H, (C 2 H 5)} 3 N + H, (C 3 H 7) 3 N + H, (HOCH 2 CH
₂ ) Ammonium ions such as ₃ N ⁺ H and C ₅ H ₆ N ⁺ H (pyridinium ions).

The diacylglycerol derivative in which X in the general formula (1) is represented by the general formula (2) can be easily produced, for example, by the following production method. That is, dimyristoylphosphatidylethanolamine, dipalmitoylphosphatidylethanolamine,
Diacylphosphatidylethanolamines having R ¹ C (═O) and R ² C (═O) acyl groups such as distearoylphosphatidylethanolamine, and D-gulonic acid-γ-lactone, gluconolactone, mannonic acid- γ
-Lactone, D-ribbonic acid-γ-lactone, α, β-glucooctanoic acid-γ-lactone, α-D-glucoheptonic acid-γ-lactone, etc. Diacylglycerol derivatives can be prepared. In this case, if necessary, trimethylamine, triethylamine, tributylamine,
Basic compounds such as amines such as pyridine and alcoholates such as sodium methoxide, sodium ethoxide, potassium methoxide and potassium ethoxide can be used.

In the above reaction, the reaction charge ratio of the diacylphosphatidylethanolamines and the saccharide lactone is not particularly limited, but the molar ratio of the diacylphosphatidylethanolamines: sugar lactone is 1:
0.5 to 1:10, preferably 1: 0.9 to 1: 1.5
Is desirable. The amount of the basic compound used is preferably equimolar or more to the diacylphosphatidylethanolamines, and the basic compound may be used as a reaction solvent.

For the reaction, chloroform, ethyl acetate, benzene, toluene, acetonitrile, tetrahydrofuran, 1,4-dioxane, dimethylformamide, dimethylacetamide, dimethylsulfoxide, triethylamine, tributylamine, tri (2-hydroxyethyl) amine, pyridine, etc. In an organic solvent and a mixed solvent thereof at a reaction temperature of −100 to + 150 ° C., preferably +30 to + 100 ° C., for 30 minutes to 200 hours, preferably 1 to 24 hours. After completion of the reaction, extraction, recrystallization, reprecipitation, adsorption treatment,
It can be isolated and purified by methods such as column treatment, ion exchange resin treatment, gel filtration, ultrafiltration, dialysis, and thin layer chromatography.

In the above production method, the reaction when D-gulonic acid-γ-lactone is used as the lactone of the saccharide is shown in the following reaction formula (4). In the formula, R ¹ C (═O) and R ² C (═O) have the same meanings as described above.

Embedded image

As another production method, an active esterifying agent is used in the presence of a dehydrating condensing agent to prepare D-gulonic acid and D-gulonic acid.
After activating aldonic acids such as gluconic acid, mannonic acid, D-ribbonic acid, α, β-glucooctanoic acid, α-D-glucoheptonic acid, dimyristoylphosphatidylethanolamine, dipalmitoylphosphatidylethanolamine, distearoylphosphatidyl The diacylglycerol derivative of the present invention can also be produced by reacting diacylphosphatidylethanolamines having an R ¹ C (═O) and R ² C (═O) acyl group such as ethanolamine.

As the dehydration condensation agent, a BOP reagent,
1,1′-carbonyldiimidazole, N-cyclohexyl-N ′-(2-morpholinoethyl) carbodiimide meso-p-toluenesulfonate, dicyclohexylcarbodiimide, diethylphosphorusanidate, N-
Examples thereof include ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride.

As the activated ester agent, N, N '
-Disuccinimidyl carbonate, 1-hydroxybenzotriazole, N-hydroxyphthalimide, isobutyl chloroformate, 4-hydroxyphenyl dimethyl sulfonium methyl sulfate, etc. are mentioned. The reaction can be carried out using the same reaction solvent as above under the same conditions.

As another production method, D-gulonic acid, D-gluconic acid, mannonic acid, D-ribbonic acid, α,
The diacylglycerol derivative of the present invention can also be produced by reacting the acid chloride of an aldonic acid such as β-glucooctanoic acid or α-D-glucoheptonic acid with the phosphatidylethanolamine.
The reaction can be carried out using the same reaction solvent as above under the same conditions.

The diacylglycerol derivative in which X in the general formula (1) is represented by the general formula (3) can be easily produced, for example, by the following production method. That is, 1,2-dimyristoyl glycerol, 1,2-dipalmitoyl glycerol, 1,2-distearoyl glycerol and the like R ¹ C (═O) and R ²
After reacting 1,2-diacylglycerols having a C (═O) acyl group with N, N′-carbonyldiimidazole (hereinafter sometimes referred to as CDI), glucamine and N-methylglucamine The diacylglycerol derivative of the present invention can be produced by reacting an amino group-containing monosaccharide such as

In the above reaction, the reaction charge of CDI is 1: 0.5 to 1:10, preferably 1: 0.9 to 1: 1, in the molar ratio of 1,2-diacylglycerols: CDI.
1.1, the amount of the amino group-containing monosaccharide charged in the reaction was
The molar ratio of diacyl glycerols: amino group-containing monosaccharides is 1: 0.5 to 1:10, preferably 1: 0.9 to 1 :.
A value of 1.5 is desirable.

The reaction is carried out in an organic solvent such as chloroform, ethyl acetate, benzene, toluene, acetonitrile, tetrahydrofuran, 1,4-dioxane, dimethylformamide, dimethylacetamide, dimethylsulfoxide and the like and a mixed solvent thereof at a reaction temperature of -100 to 100.degree. +
It can be carried out by stirring at 150 ° C., preferably +30 to + 100 ° C., for 30 minutes to 200 hours, preferably 1 to 24 hours. After completion of the reaction, extraction, recrystallization,
Reprecipitation, adsorption treatment, column treatment, ion exchange resin treatment,
It can be isolated and purified by a method such as gel filtration, ultrafiltration, dialysis, and thin layer chromatography.

In the above production method, the reaction when glucamine is used as the amino group-containing monosaccharide is shown in the following reaction formula (5). In the formula, R ¹ C (═O) and R ² C (═O) have the same meanings as described above.

Embedded image

The diacylglycerol derivative of the present invention thus obtained can be used as a membrane-forming component of liposomes. In this case, a polyvalent hydroxyl group derived from a monosaccharide can be introduced into the surface of the obtained liposome, whereby the storage stability can be improved and a high blood concentration can be maintained for a long period of time. Further, the diacylglycerol derivative of the present invention is
It can be used as a raw material for fat emulsions, medicines, agricultural chemicals, cosmetics and the like.

[0021]

The diacylglycerol derivative of the present invention is novel and useful. INDUSTRIAL APPLICABILITY The diacylglycerol derivative of the present invention can be used as a liposome membrane-forming component, and in this case, a polyvalent hydroxyl group derived from a monosaccharide can be introduced on the surface of the obtained liposome, thereby improving storage stability. In addition, it is possible to maintain a high blood concentration for a long period of time. Further, the diacylglycerol derivative of the present invention can be used as a raw material for fat emulsions, medicines, agricultural chemicals, cosmetics, and the like.

[0022]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in more detail by way of examples, but the present invention is not limited thereto. Example 1 0.5 g (723 μmol) of dipalmitoylphosphatidylethanolamine, 0.11 g (1.08 mmol) of triethylamine and 0.19 g (1.08 mmol) of gluconolactone were dissolved in 10 ml of chloroform, and the mixture was dissolved at 60 ° C. for 10 hours. It was stirred. This was washed with saturated brine, the organic layer was dried over anhydrous sodium sulfate,
By concentrating under reduced pressure, the target diacylglycerol derivative represented by the following formula (6) in the form of white powder was obtained. The obtained compound was identified by performing IR spectrum and elemental analysis after confirming that it was one spot in thin layer chromatography using chloroform / methanol (10: 1 (v / v)) as a developing solvent. did. The analysis results are as follows.

[0023]

Embedded image

IR (KBr method, cm ⁻¹ ) 3341 (Polyvalent hydroxyl group) 1737 (Carbonyl group (ester bond)) 1648 (Carbonyl group (amide bond)) 1070 (P—O—C bond) Elemental analysis (C ₄₃ H ₈₃ NO ₁₄ P) Calculated value C: 59.4%, H: 9.6%, N: 1.6% Actual value C: 59.0%, H: 10.3%, N: 1.4%

Example 2 250 mg (439 μmol) of 1,2-dipalmitoylglycerol and N, N ′ in 5 ml of chloroform.
-Carbonyldiimidazole 0.071 g (439 μm
ol) was added and the mixture was stirred at room temperature for 30 minutes. Add 0.119 g (658 μmol) of glucamine, and add 2 at room temperature.
Stir for 4 hours. This was washed with saturated saline, the organic layer was dried over anhydrous sodium sulfate, and then concentrated under reduced pressure to obtain the desired diacylglycerol derivative represented by the following formula (7) as a white powder. . The obtained compound was identified by performing IR spectrum and elemental analysis after confirming that it was one spot in thin layer chromatography using chloroform / methanol (10: 1 (v / v)) as a developing solvent. did. The analysis results are as follows.

[0026]

[Chemical 6]

IR (KBr method, cm ^-1 ) 3300 (Polyvalent hydroxyl group) 1731 (Carbonyl group (ester bond)) 1649 (Carbonyl group (amide bond)) Elemental analysis (C ₄₂ H ₈₁ NO ₁₁ ) Calculated value C: 65.0%, H: 10.5%, N: 1.8% Actual value C: 64.7%, H: 11.0%, N: 1.8%

Example 3 250 mg (439 μmol) of 1,2-dipalmitoylglycerol and N, N 'in 5 ml of chloroform.
-Carbonyldiimidazole 0.071 g (439 μm
ol) was added and the mixture was stirred at room temperature for 30 minutes. Furthermore, 0.128 g (658 μmol) of N-methylglucamine was added, and the mixture was stirred at room temperature for 24 hours. This was washed with saturated brine, the organic layer was dried over anhydrous sodium sulfate, and then concentrated under reduced pressure to obtain the desired diacylglycerol derivative represented by the following formula (8) as a white powder. . The obtained compound was identified by performing IR spectrum and elemental analysis after confirming that it was one spot in thin layer chromatography using chloroform / methanol (10: 1 (v / v)) as a developing solvent. did. The analysis results are as follows.

[0029]

[Chemical 7]

IR (KBr method, cm ^-1 ) 3298 (Polyvalent hydroxyl group) 1735 (Carbonyl group (ester bond)) 1624 (Carbonyl group (amide bond)) Elemental analysis (C ₄₈ H ₈₃ NO ₁₁ ) Calculated value C: 65.4%, H: 10.6%, N: 1.8% Actual value C: 65.2%, H: 11.1%, N: 1.7%

Reference Example Egg yolk phosphatidylcholine 20 mg (26 μmol),
Cholesterol 3.9 mg (10 μmol) and diacylglycerol derivative 2.4 mg obtained in Example 1
(10% by weight based on the total amount of the above two) was placed in an eggplant-shaped flask, dissolved in 2 ml of benzene, and then freeze-dried. To this, 1 ml of physiological saline was added, and a multi-layer liposome was obtained by bath-type ultrasonic irradiation and a vortex mixer. 3.0 by Extruder,
A large unilamellar liposome was obtained by sequentially passing through a polycarbonate membrane of 1.0 and 0.2 μm. The particle size of the obtained liposome was measured by a laser scattering particle size distribution analyzer (NIC
The particle size was 180 nm (CV value 14%) when measured using OCOM, NICOMP370HPL (trademark). After leaving the liposomes at 5 ° C. for 3 weeks,
When the particle size was measured again, it was 192 nm (CV value 18%).
It was excellent in stability.

Claims (1)

[Claims] 1. A diacylglycerol derivative represented by the following general formula (1). Embedded image (In the formula (2), M represents a monovalent counter ion, and in the formula (3), R ³ represents a hydrogen atom or a methyl group). R ¹ C (═O) and R ² C (═O) represent an acyl residue of a fatty acid having 3 to 30 carbon atoms and may be the same or different. n represents a positive number of 2 to 6. )