JPH0426699A - Amino acid derivative and production thereof - Google Patents

Abstract

NEW MATERIAL:A compound of formula I [R1-O- is cholesterol (cholesterin) or cholestanol; R2 is hydroxymethyl, alkyloxycarbonyl or lower mono or dialkylaminocarbonyl; R3 is lower aminoalkyl ]. EXAMPLE:N<epsilon>-benzyloxycarbonyllysinolcholesterolurethane. USE:An amino acid-based cationic surfactant (with high safety and good biodegradability). PREPARATION:A condensation is made between (A) a compound of formula II [R2' is carboxyl; R4 is protective group eliminable under acidic condition (reductively) for the terminal amino group of lower aminoalkyl group] and (B) a cholesterol haloformate or cholestanol haloformate of formula III(X is halogen; broken line means single or double bond).

Description

Detailed Description of the Invention (Field of Industrial Application) The present invention relates to an amino acid derivative and a method for producing the same, and more specifically, it is useful as an amino acid-based cationic surfactant that is highly safe and has good biodegradability. The present invention relates to a novel basic amino acid derivative consisting of a basic amino acid and a cholesterol or cholestanol derivative, and a method for producing the same.

(Prior Art) A surfactant is an amphipathic substance that has a parent wood group and a hydrophobic group (lipophilic group) in its molecule.

Synthetic surfactants are widely used in a wide variety of fields, but
In recent years, natural products and natural surfactants derived from natural products have attracted attention because they are safer than conventional synthetic surfactants (and are expected to have new functions and properties).

Amino acid surfactants, one of the most typical natural surfactants, are currently used in large numbers in the cosmetics field, and their types and applications are expanding further.

However, most of the ionic properties of amino acid surfactants are anionic, nonionic, or amphoteric, and there are only a few examples of cationic surfactants.

Although cationic amino acid surfactants are expected to be used for various special purposes as highly functional substances,
A few reports (N shown in JP-A No. 51-5413)
a-long chain acyl basic amino acid derivatives, Noda A
, et al., Chem, Pharm, Bull, , et al.
19.196199 (1971)), and the present invention targets urethane bonds between basic amino acids and cholesterol (cholesterin) or cholestanol derivatives. Very little is known about the amino acid steroid derivatives consisting of

On the other hand, for special applications, compounds that are currently commonly used as positively charged substances (cationic surfactants) for membrane materials of liposomes (lipid bilayer vesicles with an aqueous phase in their inner shells) are , only stearylamine, a synthetic surfactant, is available.

However, reports regarding the toxicity of stearylamine (e.g. Tyrell, D. A., et al., Bioc.
Him.

Biohs, Acta, 457, 259 (197
6), etc.), and when used in biological systems, there is a problem that sufficient attention must be paid to its safety.

Therefore, in order to solve this problem, there is currently a strong demand for highly safe and highly functional positively charged substances for liposome membrane materials.

(Problems to be Solved by the Invention) The purpose of the present invention, which was made in view of the current situation, is to provide a highly functional cationic surfactant that is highly safe and has excellent biodegradability, particularly positively charged for liposome membrane materials. provide the substance, and
The objective is to provide a method that can produce this efficiently.

(Means for Solving the Problems) The present invention, which achieves the above-mentioned objects, is characterized in that a novel amino acid steroid derivative consisting of a basic amino acid and a cholesterol or cholestanol derivative is a compound that satisfies the above-mentioned needs, and This was accomplished based on the discovery of a method for efficiently producing compounds, and the gist thereof will be explained in detail below.

That is, the present invention is based on the general formula [I]: [In the formula, R]-0- represents a cholesterol (cholesterin) or cholestanol group bonded at the 3-position. R
2 represents hydroxymethyl, alkyloxycarbonyl, or lower mono- or dialkylaminocarbonyl group. R3 represents a lower aminoalkyl group] The present invention is characterized by basic amino acid derivatives and acid addition salts thereof, and methods for producing these compounds.

Further, the general formula [■]: 11] O [wherein, R1-0- represents cholesterol (cholesterin) or cholestanol group bonded at the 3-position. R
2 represents hydroxymethyl, alkyloxycarbonyl, aminocarbonyl, or a lower mono- or dialkylaminocarbonyl group. R4 represents a lower aminoalkyl group with a protective group attached to the terminal amino group that can be removed under acidic conditions or reductively], and furthermore, the method for producing these compounds. This is a characteristic feature.

Specifically, the compound represented by the above general formula [I] is a hydroxymethyl group whose carboxyl group is a reduced form of lysine or ornithine, which is a basic amino acid, or a t-butoxycarbonyl group whose carboxyl group is its ester form, or Aminocarbonyl group, which is the amide type, or has 1 to 1 carbon atoms
It is a compound converted into a mono- or dialkylaminocarbonyl group consisting of 6 straight-chain or branched-chain alkyl groups, and its N (Z-amino group and cholesterol (cholesterin) or the double bond at the 5th and 6th positions). is a basic amino acid derivative, or an acid addition salt thereof, in which the hydroxyl group at the 3-position of reduced cholestanol is bonded with urethane through a carbonyl group.

This compound [N is an amino acid-based cationic surfactant, and can be said to be a novel natural surfactant obtained by recombining and chemically modifying natural product constituents.

Compound [II] represented by general formula [n] is a compound in which a benzyloxycarbonyl group or a t-butoxycarbonyl group, which is a protecting group for the amino group, is attached to the terminal amino group of the amino acid moiety of compound [I]. Compound [I] can be obtained by removing these protecting groups by a conventional method under reductive or acidic conditions.

The basic amino acid, which is one of the constituent parts of these compounds [I] and [II], has one amino group and one carboxyl group in the molecule, as well as an amino group, a guanidino group, and an imidazolyl group. Any basic amino acid having one or more basic substituents such as, for example, naturally derived lysine, ornithine, arginine, histidine, etc. may be used, but other amino acids with high safety may also be used. It can be used in the same way.

Basic amino acids having an asymmetric carbon atom have optical isomers, and any of the L-form, 9-form, and DL-form can be used unless there is a particular safety problem.

Among the basic amino acids used in the present invention, lysine, which has been mass-produced and is available at an extremely low cost, is one of the most suitable amino acids as a raw material amino acid.

In addition, cholesterol (cholesterin), which is one of the other constituents of compounds [I] and [TI], is one of the most representative sterins, is widely distributed in the animal kingdom, and is found at the plasma membrane of animal cells. They account for 50 mol% of all lipids, and exist in approximately the same number as phospholipids. Furthermore, cholestanol, which is also one of the constituent components, exists in small amounts in animal bodies and can be easily obtained by catalytic reduction of cholesterol (cholesterin).

-IilU The method for producing compounds represented by formulas [I] and [II] includes the general formula [ITI]: [wherein R2 is hydroxymethyl, alkyloxycarbonyl, aminocarbonyl, or a lower mono- or dialkylaminocarbonyl group] represent. R4 represents a lower aminoalkyl group with a protective group attached to the terminal amino group that can be removed under acidic conditions or reductively, and a compound represented by the general formula [IV]: In the formula, X is a halogen , and the broken line portion represents a single bond or a double bond.] By condensing with cholesterol haloformate or cholestanol haloformate, the general formula [■]: R4N-C-0-R1 [ ■I O [In the formula, R1-0- represents cholesterol (cholesterin) or cholestanol group bonded at the 3-position. R
2 represents hydroxymethyl, alkyloxycarbonyl, aminocarbonyl, or "represents a lower mono- or dialkylaminocarbonyl group. R4 represents a lower aminoalkyl group whose terminal amino group has a protecting group that can be removed under acidic conditions or reductively. The basic amino acid derivative represented by the following general formula [
11], by removing the protecting group attached to the terminal amino group of the lower alkyl group R4 in the general formula [
A compound represented by I] can be produced.

Specifically, the compound represented by the general formula [m] is a hydroxymethyl group whose carboxyl group is the reduced form of lysine or ornithine, which is a basic amino acid, a t-butoxycarbonyl group whose carboxyl group is the reduced form, or a t-butoxycarbonyl group whose carboxyl group is the reduced form, or its ester form. It is a compound converted into an amide-type aminocarbonyl group or a mono- or dialkylaminocarbonyl group consisting of a linear or branched alkyl group having 1 to 6 carbon atoms, and the Na-amino group is unsubstituted and free. However, it is a compound in which a benzyloxycarbonyl group or a t-butoxycarbonyl group, which is a protective group, is attached to the terminal amino group.

Among the basic amino acid derivatives of the compound represented by the general formula [rll], the compound in which the R2 moiety is converted to its reduced form, hydroxymethyl group, can be prepared by a known method (for example, 5h
unichi Yamada et al; Chem, Pha
rm.

Bull, 11(9), 1140-1145 (
1963) and JP-A-52-148036).

That is, first, among the basic amino acid derivatives of the compound represented by the general formula [In], an amino acid derivative in which the carboxyl group moiety is in the ester form or an acid addition salt thereof is used as a raw material compound, and a general reduction method is applied to it. Among the basic amino acid derivatives of the compound represented by the general formula [m], for example, by applying a method such as contacting with a reducing agent in an appropriate solvent,
Compounds in which the R2 moiety is substituted with a hydroxymethyl group, which is its reduced form, can be obtained.

Examples of reducing agents in this case include metal hydride compounds such as lithium borohydride, sodium borohydride, lithium aluminum hydride, sodium dihydro bis=
Examples include, but are not limited to, hydrogenated organometallic compounds such as (2-methoxyethoxy)aluminate, diborane, and the like. In addition to these reducing agents, halogen compounds such as calcium chloride can also be used as additives.

As a solvent during reduction, for example, when using sodium borohydride, solvents such as alcohols or hydrous alcohols can be used, but other solvents that do not affect the reduction reaction, such as dioxane and tetrahydrohydride, can also be used. Ethers such as furan, ethyl ether, diethylene glycol dimethyl ether, and aromatics such as benzene may also be used. In addition to these solvents, additives such as diethylamine, pyridine, aniline, N-
Amines such as methylamine and γ-collidine may be used in combination.

The reaction temperature during reduction generally proceeds at room temperature, but the reaction may be appropriately cooled or heated to advantageously proceed.

Among the basic amino acid derivatives of the compound represented by the general formula [11T], the compound in which the R2 moiety is converted to an alkyloxycarbonyl group, which is the ester type thereof, is a compound represented by the general formula [I11], as described above. Among the basic amino acid derivatives, it can be obtained and produced in the same manner as when it is used as a starting material for synthesizing a compound in which the R2 moiety is converted to its reduced form, hydroxymethyl group.

Among the basic amino acid derivatives of the compound represented by the general formula [II], the compound in which the R2 moiety is converted to an ester-type alkyloxycarbonyl group is represented by the general formula [I11]
The carboxyl group moiety which is R2' of the compound represented by is removed by a conventional method (for example, J, Am, Chem.

Yin, translation, 719 (1961);
1251 (1963), etc.), and the R2 portion consists of an aminocarbonyl group in the acid amide form thereof, or a linear or branched alkyl group having 1 to 6 carbon atoms. Alternatively, a compound converted to a dialkylaminocarbonyl group has the general formula [
It can be produced by converting the carboxyl group portion, which is R2', of the compound represented by [III] into an acid amide type by a known method.

That is, known methods include the active esterification method, mixed acid anhydride method, azide method, acid chloride method, symmetric anhydride method, and methods using coupling reagents, which are generally used for peptide bond forming reactions. . Examples of coupling reagents include N.

N'-cyclohexylcarbodiimide, carbonylimidazole, diphenylphosphoryl azide, etc., and N,N
There is a method using '-cyclohexylcarbodiimide and an additive. In this case, the additive may be, for example, N
-Hydroxysufushimide, 1-hydroxybenzotriazole, N-hydroxy-5-norbornene-92,
Examples include 3-dicarboxylic acid imide. Examples of the solvent in this case include alcohols such as methanol and ethanol, halogenated hydrocarbons such as chloroform and dichloromethane, ethers such as tetrahydrofuran and ether, N,N-dimethylformamide, and pyridine. , but not particularly limited to these.

The compound represented by the general formula [rV] is specifically a haloformate of cholesterol (cholesterin) or cholestanol, and can be prepared by a known method (for example, H, Wie
land et al.; Z, Ph 5io1. Engineering hem.

(1923) etc.). It is also easily available as a commercially available product such as cholesterol chloroformate.

The halogen represented by X in the compound represented by the general formula [IV] is not particularly limited, but chlorine is generally used.

In order to obtain the compound represented by the general formula [11] by condensing the compounds represented by the general formulas [III] and [IV], both of them are combined in a suitable solvent under basic conditions by a conventional method. It can be done by reacting below.

Examples of solvents in this case include alcohols such as methanol and ethanol, halogenated hydrocarbons such as chloroform and dichloromethane, ethers such as tetrahydrofuran, ether and dioxane, aromatics such as benzene and toluene, and even N, N-dimethylformamide,
Examples include pyridine, but the solvent is not particularly limited to these, and other solvents that do not affect the condensation reaction may be used.

Examples of bases used to make basic conditions include tertiary amines such as trimethylamine, triethylamine, and pyridine; hydration of alkali metals such as sodium and potassium; and alkaline earth metals such as magnesium and calcium. Examples include carbonates, carbonates, hydrogen carbonates, etc., but are not particularly limited thereto.

Furthermore, as for the reaction temperature, this condensation reaction usually proceeds at room temperature, but the reaction may be suitably cooled or heated to advantageously proceed.

In order to purify the target product from the reaction station, general recrystallization or column chromatography can be applied. For example, after the completion of the above condensation reaction, the reaction solution is washed with water, the organic phase is extracted, an appropriate drying agent (for example, anhydrous sodium sulfate, anhydrous magnesium sulfate, etc.) is added thereto, and after drying, under reduced pressure, The solvent is distilled off, and the residue is purified by recrystallization in a conventional manner or by elution with a suitable mixed solvent using a silica gel column or the like.

As described above, the compound represented by the general formula [11] can be obtained, but apart from this method, the compound represented by the general formula [II
Among the compounds represented by I, compounds in which the R2 moiety is converted into an aminocarbonyl group in its amide form, or a mono- or dialkylaminocarbonyl group consisting of a linear or branched alkyl group having 1 to 6 carbon atoms are , also produced by the method described below.

That is, the general formula [111'] [wherein R2' represents a carboxyl group]. R4 represents a lower aminoalkyl group with a protective group attached to the terminal amino group that can be removed under acidic conditions or reductively] and cholesterol haloformate represented by the general formula [IV] or By condensing with cholestanol haloformate, a compound of the general formula [11'] [wherein R1-0- represents cholesterol (cholesterin) or cholestanol group bonded at the 3-position] is obtained. R
2' represents a carboxyl group. R4 represents a lower aminoalkyl group with a protective group attached to the terminal amino group that can be removed under acidic conditions or reductively] After obtaining the amino acid derivative represented by the general formula [■' R2 ' into an alkyloxycarbonyl, aminocarbonyl, or lower mono- or dialkylaminocarbonyl group,
A compound represented by the general formula [11] (however, in this case, R
2 represents only an alkyloxycarbonyl, aminocarbonyl, or lower mono- or dialkylaminocarbonyl group).

That is, specifically, the carboxyl group of the terminal amino group protecting group basic amino acid of R4 in the compound represented by the general formula [rll'l is in a free state.

The compound represented by the general formula [ITI'] and the general formula [T
V] is subjected to a condensation reaction in the same manner as above to obtain a compound represented by the general formula [■'], and then
By esterifying or amidating the carboxyl group of the compound represented by the general formula [■'1] as described above, the compound represented by the general formula [11] (
However, in this case, R2 may represent only alkyloxycarbonyl, aminocarbonyl, or a lower mono- or dialkylaminocarbonyl group).

In this case, the conditions for the condensation reaction, esterification or amidation reaction, purification step, etc. are based on the general formula [III]
The conditions for producing the compound of [III] can be similarly applied (however, in this case, the reaction proceeds more favorably if caustic soda or the like is used as the base during the condensation reaction, rather than, for example, triethylamine or the like).

A benzyloxycarbonyl group or a t-butoxycarbonyl group, which is a protecting group attached to the terminal amino group of the compound represented by the general formula [III, obtained as above, is removed by a conventional method under reducing or acidic conditions. By elimination, a compound represented by the general formula [ff] can be obtained.

The reaction for removing the protecting group can be carried out by a known method for peptide synthesis.

That is, for example, the reductive deprotection method includes a catalytic reduction method.

Examples of catalysts used for catalytic reduction include platinum chloride,
Examples include platinum catalysts such as platinum oxide, palladium catalysts such as palladium, palladium on carbon, catalysts such as nickel oxide, Raney nickel, and catalysts such as Raney cobalt.The solvent for catalytic reduction is usually water. , methanol, ethanol, propatool, N,N-dimethylformamide, acetic acid, formic acid, or a mixed solvent thereof.

In addition, examples of acids used in the deprotection method under acidic conditions include trifluoroacetic acid, hydrochloric acid, formic acid, and hydrogen fluoride, and examples of solvents in this case include dichloromethane, chloroform, dioxane, and tetrahydrofuran. , alcohols, acetic acid, etc., but are not particularly limited to these.

Regarding the reaction temperature during deprotection, the reaction usually proceeds at room temperature, but the reaction can be advantageously progressed by cooling or heating as appropriate.

Examples of the acid addition salt of the compound represented by the general formula [I] obtained as above include organic acid salts such as formate, acetate, trifluoroacetate, maleate, tartrate, and oxalate. Examples include addition salts, and inorganic acid addition salts such as hydrochloride, sulfate, hydrobromide, and nitrate, but are not particularly limited thereto.

The acid addition salt of the compound represented by the general formula [T] can be used by removing the addition salt by a conventional method and returning it to a free form, if necessary.

The compound represented by the general formula [I] is a natural amino acid derivative obtained by recombining and chemically modifying amino acids and cholesterol derivatives, which are components of natural products, and is highly safe and has excellent biodegradability. It can be said to be an amino acid-based cationic surfactant.

Moreover, since the compound represented by -formation [I] has an amino acid and a cholesterol derivative bonded with urethane, it is safe and stable, and is advantageous when used as a raw material for formulation.

Many reports (for example, JP-A-58-134197, etc.)
As mentioned above, amino acid higher alkyl ester salts and their N-lower alkyl substituted products have a common drawback of ester salts, as they have an ester bond in their structure and are therefore highly hydrolyzable, making it difficult to stably incorporate them into products. It is said that it is often difficult to do so.

Furthermore, for special purposes, there have been reports on the use of amino acid derivatives as liposome-forming materials (e.g.,
No. 2921, etc.). In this report, an amino acid-based anionic surfactant, which is an N-higher acylamino acid, is used as a lipid that is easily degradable in the living body, has low toxicity, and has the ability to form liposomes.

In addition, there are reports of attempts to target cancer cells by modifying liposomes with amino acid lipid derivatives (e.g., 7th Biopharmaceutical Research Society Abstracts, p. 49-(1989), etc.). Na-cocoyl-arginine ethyl ester (CEA), an amino acid-based cationic surfactant
We report on the new properties of liposomes whose membranes are modified with amino acids.

However, all of the amino acid derivatives used for these belong to amino acid acyl derivatives, and when these are applied to living organisms, there are concerns about safety issues such as high hemolysis.

On the other hand, reports on the surface modification of liposomes with cholesterol derivatives having amine groups (e.g., K, R,
Patel et al., Biochimica et Bio
h 5ica drunkenness” for Yu, 256-264 (19
85), etc.), but no reports regarding amino acid steroid derivatives such as those of the present invention have been found.

The constituent parts of liposomes are generally divided into three types:
That is, various phospholipids as a base material, cholesterol as a membrane-strengthening substance, and various cationic or anionic surfactants as a membrane-charged substance are often used.

In addition, by adding a charged substance to the liposomes and imparting a surface charge, the aggregation of liposomes is prevented, and the inner aqueous phase is expanded by electrostatic repulsion between the lamellar layers, allowing water-soluble encapsulated substances to form in the liposomes. It is known that the behavior of liposomes in vivo (for example, tissue migration) is greatly influenced by the type of charged substance used while increasing the amount of encapsulated and retained substances.

When the amino acid-based cationic surfactant of the present invention is used as a liposome membrane material, its chemical structure allows it to act as both a positively charged substance and cholesterol, which is a membrane-strengthening substance.

(Effects of the Invention) As is clear from the above explanation, according to the present invention, as a novel natural amino acid derivative, (1) it is highly safe, has excellent biodegradability, and combines amino acids and cholesterol derivatives; Because it is urethane-bonded, (2) it can be stably incorporated into products (formulations).

In addition, when used as a liposome membrane material, it can be used as (3) a safe and novel membrane-charged substance, (4) a membrane-charged substance with a structure that also serves as a membrane-strengthening substance, and (5) a membrane for amino acid-modified liposomes. As a substance,
It is possible to provide an amino acid-based cationic surfactant that imparts unprecedented properties to liposomes and has advantages such as stiffness.

Therefore, it is an amino acid derivative suitable for use in fields such as medical liposomes, which require consideration of biocompatibility and various affinities with biomembranes, as well as biomedical materials,
It is expected to be applied to cosmetics, etc., and is also expected to be used in a wide range of fields such as biochemistry, medicine, pharmacy, and engineering, and has great industrial value.

(Example) Next, the present invention will be explained in more detail with reference to Examples.
It is not limited to these.

Reference example, N (Synthesis of Z-amino group protected ricinol N
Dissolve 3.31 g (10 m mon) of 8-benzyloxycarbonyllysine methyl ester hydrochloride (Sigma) in 100 mρ of ethanol, and add 2 ml of ethanol under cooling with water.
Sodium borohydride suspended in 0 mj2 2.66
g (70 m moβ) was added dropwise. After the dropwise addition was completed, the mixture was refluxed in an oil bath for one day. The reaction solution was filtered, the solvent was distilled off under reduced pressure, and the concentrate was redissolved in chloroform and washed with water. After drying the organic layer over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and the resulting crude product was crystallized in acetonitrile to obtain 2.05 g of the target product, N8-penzyloxycarbonylridinol ( yield 77%).

Example 1 Synthesis of N8-penzyloxycarbonylridinol cholesterol urethane Cholesterol roformate (Tokyo Kasei Co., Ltd.) 2.87 g
(6,93m mo,e) and 1.69 g of N8-penzyloxytriponylridinol (6,35m mo °C)
was dissolved in dichloromethane (100 m.rho.), triethylamine (1.8 mJ2 (12.9 m.rho.)) was added thereto, and the mixture was stirred at room temperature for 6 hours. The reaction solution was washed with water, and the organic layer was dried over anhydrous magnesium sulfate and then concentrated to dryness under reduced pressure. The resulting residue was applied to a silica gel column and eluted with a mixed solvent of n-hexane and ethyl acetate (1:1). The target fractions were collected using thin layer chromatography (hereinafter abbreviated as TLC) for the obtained elution fractions, and after concentration under reduced pressure to dryness, 3.55 g of solid matter was obtained.
(yield 82%). By crystallizing this solid substance in methanol, the target product, N8-penzyloxylponylridinol cholesterol urethane 3.02
I got g.

Example 2 Synthesis of N8-benzyloxycarbonyl lysine cholesterol urethane Cholesterol dilorformate (Tokyo Kasei Co., Ltd.) 6.51 g
(14,5 m moI2) and N8-benzyloxycarbonyl lysine (Tokyo Kasei Co., Ltd.) 4.09 g (14,6
mmoβ) was dissolved in 150 mρ of chloroform, and while stirring at room temperature, 14% of 2N sodium hydroxide was added to this solution.
mβ was added in two portions, and the mixture was stirred at room temperature for 6 hours. A small amount of methanol was added to the reaction solution, and the reaction solution was acidified by dropping an appropriate amount of diluted hydrochloric acid, followed by extraction with a mixed solvent of methanol-chloroform (1:10). The extracted organic layer was dried over anhydrous magnesium sulfate, concentrated to dryness under reduced pressure, and the resulting residue was applied to a silica gel column and eluted with a methanol-chloroform (1:20) mixed solvent. The obtained target fraction was concentrated to dryness under reduced pressure to obtain N8 benzyloxycarbonyl lysine cholesterol urethane solid substance 7.
．． 43 g (yield 74%) was obtained.

Example 3 Synthesis of N8-benzyloxycarbonyldiethylamide lysine cholesterol urethane N8 synthesized in Example 2
- 2.55 g (3.6 mmol) of benzyloxycarbonyl lysine cholesterol urethane was dissolved in 50 mA of tetrahydrofuran, and 570 g of diethylamine was dissolved in it.
μj2 (5.5 mmo) was mixed with 0.94 g of hydroxybenztriazole (6,1 mmoβ) and 1.51 g of dicyclohexylcarbodiimide (7,3
mmoj2) was added, and the mixture was stirred for 2 hours under water cooling, and then stirred for 1 day at room temperature. After filtering the reaction solution, the filtrate was concentrated to dryness under reduced pressure.

The obtained solid was dissolved in a small amount of ethyl acetate, filtered again, and the filtrate was washed with a saturated aqueous solution of thorium bicarbonate.
After washing twice with water, the organic layer was dried over anhydrous magnesium sulfate and concentrated to dryness under reduced pressure. The resulting residue was applied to a silica gel column and eluted with a mixed solvent of n-hexane and ethyl acetate (2:1).

The obtained target fraction was concentrated to dryness under reduced pressure to obtain 2.29 g (yield: 85%) of N8-benzyloxycarbonyl diethylamide lysine cholesterol urethane solid.

Example 4 Synthesis of ricinol cholesterol urethane 172 mg of 10% palladium on carbon (Kojima Chemical Co., Ltd.) was mixed with 1 part of ethanol.
00 mρ, and to this suspension 1.3
6g (2,0mmolI2) was added and placed in an atmospheric hydrogenation apparatus (
(manufactured by Hoben Shokai) for 6 hours at room temperature. The reaction solution was filtered, and the filtrate was concentrated to dryness under reduced pressure to obtain a crude product of 1.083
g (yield 99%) was obtained. This crude product was crystallized in ethanol to obtain 0.50 g of the target ricinol cholesterol urethane.

The analysis results are shown below.

R1: Cholesterin N.

1, CH20H 2, CH20H 3, C00t-Bu 4, CONH2 5, CONEt 2 B, C0NEt 2 (CH2)4NH2 (CH2)3NH2 (CH2), NO3 (CH,14NH2 (CH,), NO3 (CH2)3NH2 Compound Physicochemical data of mp: ("C) IR: (am"') 'HNMR: (ppm, CDCLi) MS:
(m/z, M”) No. 1: 168-169 MR S: 3370, 1695, 1560, 1470.138
0,1255: 0.67 (3H, s), 0.78
~2.08 (358, m).

(6H, d), 0.88 (3H, d), 0.9
7 (3H, s).

2.13-2.42(2)(,m), 2.67(2
H,t).

3.47-3.72 (3H, m), 4.46 (LH
,br).

4.87 (LH, br), 5.36 (IH, b
r): 545 0.83 No, 2 = 187~19O MR S: 3340, 1695, 1540, 1380.125
5: 0.67 (3H, s), 0.74-2.08
(33H, m).

(6H, d), 0.88 (3H, d), 0.9
7 (3H, s).

2.13-2.42 (2H, m), 2.67 (2H
,t).

3.47-3.72 (3H, m), 4.46 (18
,br).

4.87 (IH, br), 5.36 (IH, b
r) = 531 0.83 No, 3 MR: 165-16g: 3370.1730-1680.1525.147
0.1355.1205: 0.59 (3H, s)
, 0.72-2.00 (34H, m), 0.7
7 (6H, d), 0.82 (3H, d), 0.
92 (3H, s).

1.38 (9H, s), 2.17-2.32 (2H
, m).

MS 2.76 (2H, t), 4.03 (IH, br)
, 4.36 (LH, br) , 5.28 (
IH, br) = 615 No, 4 mp: Amorphous IR: 3405.1705.1660.1610.1
54ONMR: 0.65 (3H, s), 0.80~
2.00 (34H, m), 0.84 (6H, d)
, 0.92 (3H, d), 1.00 (3H, s).

2.13-2.42 (2H, m), 2.70 (2H,
t) , 4.13 (IH, br) , 4.47 (IH
,br), 5.22(LH,br).

5,36 (LH,br), 5.42 (IH,b
r), 6.35 (LH, br); 558 S No, 5 NMR S: 72-74: 3450, 1705, 1635, 1540, 14
70.1250: 0.64 (3H, s), 0.8
0 to 2.05 (34H, m), 0.82 (6H, d
) , 0.87 (3H, d) , 0.97 (3H,
s), 1.08(3H,t), 1.19(3H
,t) ,2.15~2.42(2H,m) ,2
．． 70 (2H, t), 3.10-3.60 (4H,
m), 4.32 to 4.64 (2H, m), 5.
35 (LH, br), 5.50 (IH, br)
= 614 No, 6 mp: 82-84 IR: 3425 1715, 1635, 1540,
1470.125ONMR: 0.64 (3H, s)
, 0.80-2.05 (32H, m), 0.
82 (6H, d), 0.87 (3H, d), 0
．． 97 (3H, s) , ], 08 (3H, t)
, 1.19 (3H, t) , 2.15-2.42 (2
H, m), 2.70 (2H, t), 3.10~
3.62 (4H, m), 4.32-4.64 (2H
, m) , 5.36 (IH, br) , 5.50
(IH, br) MS: 600 N-C-0-R1 O R1: Cholesteryl N.

H20H CH,OH OOH OOH OOH 0OH COOt-Bu [Ir] (CH2)4NH-CBZ (CH2), NH-CBZ (CH2)4NH-CBZ (CH2)4NH-BOC (CH2) 3NH-CBZ (C:H,), NH-BOC (CH2)4NH-CBZ (:00t-Bu CON　I□ C0NH.

C0NEt.

0NEt2 CONET.

0NEt2 (C:Hg), NH-BOC (CH2)4NH-CBZ (CHg)JH-BOC: (CH2)4NH-CBZ (CH2)4NH-BOC ((:Hg)aNH-CBZ (CH,13NH-BOC compound Physicochemical data of mp: ('C) IR: (cm"') H-NMR: (ppm) No, 7 NMR = 161-163: 3440, 1710, 1695, 1540.12
60 ((:DCL*): 0.67(3)1.s)
, 0.78-2.08 (33H, m), 0.
88 (6H, d), 0.88 (3H, d)
.

0.97 (3H, s), 2.1:3-2.42 (2
)1. m) , 3.19 (2H, br) , 3.6
0 (3H, br), 4.45 (IH, br).

4.73-4.95 (2H, m), 5.06 (2H
,s).

5.34 (IH, br), 7.33 (5H, 5)
No. 8: 130-131 NMR: 3440, 1695, 1540.1265 (CD
Cl2): 0.6g (3H,s), 0.83
~2.08 (31H, m), 0.86 (6H, d)
, 0.91 (3H, d).

1.00 (3H, s), 2.17-2.43 (2H
, m) , 3.22 (2t1.br) , 3.62
(3H, br), 4.48 (LH, br).

4.8f) -4,97 (2H, m), 5.10 (2H,
s).

5,37 (LH,br), 7.35 (5H,
5) No. 9 NMR, amorphous: 3350.1710-1695.1540.125
5 (CDCL*+CD, 100): 0.63 (3
H, s), 0.74-2.06 (32H, m),
0.82 (68, d), 0.87 (3H, d).

0.95 (3H, s), 2.13-2.40 (2H
,m) ,3.13(2H,br) ,4.18(
LH, br), 4.41 (IH, br).

5.04 (2H, s), 5.31 (18, br)
, 7.29 (5H, 5) No, 1 O NMR: Amorphous + 3350.1720 to 1695.1520.126
0 (CDCL3+CD30D): 0.63 (3H
, s), 0.73-2.05 (32H, m),
0.83 (6H, d), 0.87 (3H, d).

0.96 (3H, s), 1.38 (9H, s), 2
．． 12-2.38 (2H, m), 3.01. (2H
,br), 4.12(IH,br).

4,42 (LH,br), 5.32 (IH,
br) No. 1 1 NMR: Amorphous: 3330.1715-1705.1540.126
0(CD(:L-+CDaOD): 0.63(3
H,s), 0.74-2,06 (30)1. m)
, 0.82 (6H,d) , 0.87 (3H
,d)0.95(3H,s) ,2.13~2.40(
2B, m) 3, 13 (2)1. br), 4.1
8 (IH, br).

4.4], (IH,br), 5.04(2H,s)
5.32 (IH, br), 7.29 (5H, 5) No.
, 12 NMR: Amorphous: 3350.1720-1695.1520.125
0(CDCLa+(:D30D): 0163(3
H, s), 0.73-2.05 (30H, m)
, 0.83 (6H, d), 0.87 (3H, d).

0.96(3t(,s), 1.38(9H,s)
, 2.12-2.38 (2H, m) , 3.01 (
2H,br), 4.12(II(,br).

4.42 (IH, br), 5.32 (LH, br)
No. 1 3 NMR: Amorphous: 3340.1695.1535.1455.125
0(C:DCL,): 0.65(3H,s)
,0.78~2.07(32t(,m),0.84
(6H, d), 0.89 (3H, d).

0.98 (3H, s), 1. ．． 43 (9H, s),
2.16-2.42 (2H, m), 3.17 (2H,
br) , 4.18 (LH, br) 4.46 (LH,
br), 4.79 (IH, br).

5.02-5.20 (IH, m), 5.07 (2
H, s) 5.34 (LH, br), 7.33 (
5H, 5) No, 1 4 = 140-141 NMR: 3370, 1720, 1705, 1,525.1
370 (CDC: L3): 0.65 (3H, s)
, 0.80-2.05 (32H, m) , 0.86 (
6H, d), 0.88 (3H, d) 0.98 (3H
,s) ,1.41(9H,s) ,1.43(9
1(,s)2.18-2.41i2H,m),3.
08 (2H, br).

4.16 (IH, br), 4.36-4.64
(2H, m).

5.14 (LH, br), 5.34 (LH, b
r) No, 15 NMR = 157-160: 3340, 1695, 1535, 1455.12
55 (CDC:L3): 0.68 (3H, s)
, 0.73-2.08 (32H, m), 0.86
(6H, d), 0.91 (3H, d).

0.99 (3H, s), 2.15-2.43 (2H
, m).

3.20 (2H, br), 4.12 (LH, br)
, 4.49 (IH, br), 4.82 (I
H, br), 5.10 (2H, s) 5.26-5.
37 (3H, m), 6.21 (IH, br).

7.33 (5H, 5) No, 1 6 p: 152-155 IR: 3340, 1695, 1535, 1455.
1255NMR (CDCLs): 0.65 (3H
, s), 0.80-2.10 (32H, m), 0.84
(6H, d), 0.90 (3H, d).

0.98 (3H, s), 1.44 (9H, s)
, 2.22-2.43 (2H, m), 3.10 (2
H, br), 4.12 (LH, br).

4.47 (IH, br), 4.63 (LH, b
r), 5.30-5.43 (3H, m), 5.
53 (IH, br), 6.24 (LH, b
r) No, 1 7 NMR: Amorphous: 3320, 1715, 1635, 1530.146
01245 (CDCL3): 0.65 (3H, s
), 0.78-2.05 (32H, m), 0.82
(3H, d), 0.87 (3H, d).

0.97 (3H, s), 1.08 (3H, t), 1
．． 19 (3H, t) 2.15-2.42 (2H, m)
, 3.08-3.58 (6H, m) 4.43 (IH,
br) , 4.65 (IH, br) , 4.91 (
IH,br), 5.06(2H,s), 5
．． 35 (IH, br) 5.45 (LH, br), 7
．． 33 (5H,5) No, 18 NMR: Amorphous: 3320.1715.1635.1520.146
5.1250 (CDCL, ): 0.66 (3H,
S), 0.79-2.10 (32H, m), 0.
85 (6H, d), 0.90 (3H, d).

0.99 (3H, s), 1.10 (3H, t)
, 1.21 (3H, t) 1.42 (9H, s) , 2
．． 16-2.46 (2H, m).

3.05-3.62 (6H, m), 4.34-4.7
5 (3B, m) 5.37 (IH, br), 5.47
(LH, 5) No, 1 9. 122-125 NMR: 3325,1?15,1635,1530,14
65.1250 (CDCLs): 0.63 (3H
, s), 0.78-2.05 (30H, m), 0.
82 (6H, d), 0.87 (3H, d).

0.97 (3H, s), 1.08 (3H, t)
, 1.19 (3H, t).

2.15-2.42 (2H, m), 3.05-3.
58 (6H, m).

4.43 (IH, br), 4.65 (IH, br), 4
．． 91 (IH, br), 5.06 (IH, s), 5.3
5 (18, br).

5.45 (IH, br), 7.33 (5H, 5
) No, 20: Amorphous NMR: 3325, 1710, 1640, 1510, 138
5.1250 (CDCL3): 0.66 (3H,
s), 0.79-2.10 (30H, m),
0.85 (6H, d), 0.90 (3H, d
).

0.99 (3H, s), 1.10 (3H, t)
, 1.21 (3H, t) 1.42 (9H, s) , 2
．． 16-2.46 (2H, m).

Claims (1)

[Claims] 1. General formula [I]: ▲There are mathematical formulas, chemical formulas, tables, etc.▼[I] [In the formula, R1-O- is cholesterol (cholesterin) or cholesterol bonded at the 3-position. Represents a stanol group. R
2 represents hydroxymethyl, alkyloxycarbonyl, or lower mono- or dialkylaminocarbonyl group. R3 represents a lower aminoalkyl group] Basic amino acid derivatives and acid addition salts thereof. ▲There are mathematical formulas, chemical formulas, tables, etc.▼ [II] [In the formula, R1-O- represents cholesterol (cholesterin) or cholestanol group bonded at the 3-position. R
2 represents hydroxymethyl, alkyloxycarbonyl, aminocarbonyl, or a lower mono- or dialkylaminocarbonyl group. R4 represents a lower aminoalkyl group with a protective group attached to the terminal amino group that can be removed under acidic conditions or reductively]. 3. In claim 1, R2 is a hydroxymethyl group, R
Basic amino acid derivatives in which 3 is 4-aminobutyl or 3-aminopropyl, and acid addition salts thereof. 4. The basic amino acid derivative and acid addition salt thereof according to claim 1, wherein R2 is a t-butoxycarbonyl group, and R3 is a 4-aminobutyl or 3-aminopropyl group. 5. In claim 1, R2 is an aminocarbonyl group, or a linear or branched mono- or dialkylaminocarbonyl group having 1 to 6 carbon atoms, and R3 is a 4-aminobutyl or 3-aminopropyl group. Basic amino acid derivatives and acid addition salts thereof. 6. In claim 2, R2 is a hydroxymethyl group, R
A basic amino acid derivative in which 4 is 4-aminobutyl or a 3-aminopropyl group with a protecting group attached to the terminal amino group that can be removed under acidic conditions or reductively. 7. In claim 2, R2 is a t-butoxycarbonyl group, R3 is 4-aminobutyl, or a 3-aminopropyl group with a protecting group attached to the terminal amino group that can be removed under acidic conditions or reductively. A basic amino acid derivative that is 8. In claim 2, R2 is an aminocarbonyl group, or a mono- or dialkylaminocarbonyl group consisting of a linear or branched alkyl group having 1 to 6 carbon atoms, and R4 is a 4-aminobutyl or 3-aminopropyl group. A basic amino acid derivative that has a terminal amino group attached with a protective group that can be removed under reductive or acidic conditions. 9. The basic amino acid derivative according to any one of claims 2 and 6 to 8, wherein the protecting group for the terminal amino group of the lower aminoalkyl group represented by R4 is a benzyloxycarbonyl group or a t-butoxycarbonyl group. 10. General formula [III]: ▲Mathematical formulas, chemical formulas, tables, etc.▼[III] [In the formula, R2 represents hydroxymethyl, alkyloxycarbonyl, aminocarbonyl, or a lower mono- or dialkylaminocarbonyl group. R4 represents a lower aminoalkyl group with a protective group attached to the terminal amino group that can be removed under acidic conditions or reductively] and the general formula [IV]: ▲ Numerical formula, chemical formula, table, etc. ▼ [IV] By condensing with cholesterol haloformate or cholestanol haloformate, which is represented by [In the formula, X represents a halogen and the broken line portion represents a single bond or double bond], general Formula [II]: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ [II] [In the formula, R1-O- represents cholesterol (cholesterin) or cholestanol group bonded at the 3-position. R
2 represents hydroxymethyl, alkyloxycarbonyl, aminocarbonyl, or a lower mono- or dialkylaminocarbonyl group. R4 represents a lower aminoalkyl group with a protective group attached to the terminal amino group that can be removed under acidic conditions or reductively. Method. 11. The following general formula [I], which is characterized by removing the protecting group attached to the terminal amino group of the lower alkyl group that is R4 in the compound represented by the general formula [II] produced in claim 10: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ [I] [In the formula, R1-O- represents cholesterol (cholesterin) or cholestanol group bonded at the 3-position. R
2 represents hydroxymethyl, alkyloxycarbonyl, or lower mono- or dialkylaminocarbonyl group. 2. The method for producing a compound according to claim 1, characterized in that R3 represents a lower aminoalkyl group. 12. General formula [III']: ▲Mathematical formulas, chemical formulas, tables, etc.▼[III'] [In the formula, R2' is a carboxyl group, and R4 is a lower aminoalkyl group under acidic or reductive conditions. By condensing a compound represented by [representing a compound with a removable protecting group] with cholesterol haloformate or cholestanol haloformate represented by general formula [IV], the compound represented by general formula [II'] ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ [II'] [In the formula, R1 represents cholesterol (cholesterin) or cholestanol group bonded at the 3-position. R2' represents a carboxyl group. R4 represents a lower aminoalkyl group with a protective group attached to the terminal amino group that can be removed under acidic conditions or reductively] After obtaining the amino acid derivative represented by the general formula [II'], R in the compound
The 2' carboxyl group is converted to an alkyloxycarbonyl, aminocarbonyl, or lower mono- or dialkylaminocarbonyl group to form a compound represented by the general formula [II] (however, in this case, alkyloxycarbonyl, aminocarbonyl, or lower mono- or dialkyl 3. The method for producing a compound according to claim 2, characterized in that the compound (representing only a carbonyl group) is produced. 13. The compound represented by general formula [I] is obtained by removing the protecting group attached to the terminal amino group of the lower alkyl group that is R4 in the compound represented by general formula [II] produced in claim 12 ( 2. The method for producing a compound according to claim 1, wherein R2 represents only an alkyloxycarbonyl, aminocarbonyl, or lower mono- or dialkylaminocarbonyl group. 14. In general formula [I] or [II], R1-O-
is bound at the 3rd position (cholesterin)
By hydrogenating a compound representing the group, the general formula [
3. The method for producing a compound according to claim 1 or 2, characterized in that a compound representing a cholestanol group in which R1-O- is bonded at the 3-position in [I] or [II] is produced. 15. The manufacturing method according to claim 10 or 12, wherein the halogen of X in general formula [IV] is chlorine or bromine.