JP2861262B2 - Amino acid derivative and method for producing the same - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to an amino acid derivative and a method for producing the same. Specifically, the present invention is useful as a highly safe and biodegradable amino acid-based cationic surfactant. And a novel basic amino acid derivative comprising a basic amino acid and a cholesterol or cholestanol derivative, and a method for producing the same.

(Prior Art) An amphiphilic substance having a hydrophilic group and a hydrophobic group (lipophilic group) in one molecule is a surfactant.

Synthetic surfactants are widely used in a wide variety of fields, but in recent years biosurfactants that are safer than conventional synthetic surfactants and are expected to have new functions and properties, that is, derived from natural products and natural products Natural surfactants are receiving attention.

The most typical amino acid surfactants among these natural surfactants are already used in the field of cosmetics at present, and their types and uses are becoming more widespread.

However, the ionicity of amino acid surfactants is
Most are anionic, nonionic or amphoteric, and there are only a few examples of cationic ones.

Although cationic amino acid-based surfactants are expected to be used for various special uses as highly functional substances, there are few reports (N ^α disclosed in JP-A-51-5413).
Long chain acyl basic amino acid derivatives, Noda A. et al., Che
m. Pharm. Bull. , 19 , 196-199 (1971), and the like, and there are only basic amino acids and cholesterol (cholesterol) as the target of the present invention. ) Or an amino acid steroid derivative comprising a urethane bond with a cholestanol derivative is hardly known.

On the other hand, compounds used as specially charged substances (cationic surfactants) currently used in membrane materials for liposomes (lipid bilayer vesicles having an aqueous phase in the inner shell) There is only a synthetic surfactant, stearylamine.

However, reports on the toxicity of this stearylamine (eg, Tyrell, DA et al., Biochim . Biophys . Act.
a. , 457 , 259 (1976)), and there is a problem that it is necessary to pay sufficient attention to its safety when used in biological systems.

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

(Problems to be Solved by the Invention) An object of the present invention made in view of the present situation is to provide a highly functional cationic surfactant having high safety and excellent biodegradability, in particular, a positively charged cationic surfactant for a liposome membrane material. An object of the present invention is to provide a substance and a method capable of efficiently producing the substance.

(Means for Solving the Problems) According to the present invention for achieving the above object, a novel amino acid steroid derivative comprising a basic amino acid and a cholesterol or cholestanol derivative is a compound satisfying the above-mentioned demands, and It was made based on finding a method for efficiently producing a compound,
Hereinafter, the gist will be described in detail.

That is, the present invention provides a compound represented by the general formula [I]: [In the formula, R 1 —O— represents a cholesterol (cholesterol) or cholestanol group bonded at the 3-position. R2 represents a hydroxymethyl, alkyloxycarbonyl, or lower mono- or dialkylaminocarbonyl group. R3 represents a lower aminoalkyl group], an acid addition salt thereof, and a method for producing these compounds.

Also, the general formula [II]: [In the formula, R 1 —O— represents a cholesterol (cholesterol) or cholestanol group bonded at the 3-position. R2 represents a hydroxymethyl, alkyloxycarbonyl, aminocarbonyl, or lower mono- or dialkylaminocarbonyl group. R4 represents a terminal amino group of a lower aminoalkyl group having a protective group capable of leaving under reductive conditions or under acidic conditions]], and a method for producing these compounds. It is a feature.

The compound represented by the general formula [I] is specifically a basic amino acid such as lysine or ornithine, a carboxyl group of which is a reduced form of a hydroxymethyl group, an ester form of which is a t-butoxycarbonyl group, or An amide-type aminocarbonyl group or a compound having 1 carbon atom
A 6 straight-chain or compound that is converted to a mono or dialkylamino group consisting of branched alkyl groups, the N ^alpha - amino group and cholesterol (cholesterol) or 5-position, 6-position of the double bonds Is a basic amino acid derivative in which a hydroxyl group at the 3-position of reduced cholestanol is urethane-bonded via a carbonyl group between them, or an acid addition salt thereof.

This compound [I] 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 components.

The compound [II] represented by the general formula [II] is a compound wherein the terminal amino group of the amino acid portion of the compound [I] has a benzineoxycarbonyl group or a t-butoxycarbonyl group as an amino-protecting group. Compound [I] can be obtained by removing these protecting groups by a conventional method under reducing or acidic conditions.

The basic amino acid which is one of the constituent parts of the 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 at least one basic substituent such as lysine, ornithine, arginine, histidine, etc. may be used, but other highly safe amino acids may also be used. Can be used as well.

There are optical isomers of basic amino acids having an asymmetric carbon atom.
Both body and DL body can be used.

Among the basic amino acids that can be used in the present invention, lysine, which has been mass-produced and can be obtained at extremely low cost, is one of the most suitable as a starting amino acid.

Cholesterol (cholesterol), one of the other components of compounds [I] and [II], is one of the most typical sterins and is widely distributed in the animal kingdom, Occupies 50 mol% of the total lipids and is present in approximately the same number as the phospholipids. In addition, cholestanol, which is also one of the components, is slightly present in the animal body and can be easily obtained by catalytic reduction of cholesterol (cholesterol).

The method for producing the compounds represented by the general formulas [I] and [II] is as follows: [In the formula, R2 represents a hydroxymethyl, alkyloxycarbonyl, aminocarbonyl, or lower mono- or dialkylaminocarbonyl group. R4 represents a terminal amino group of a lower aminoalkyl group having a protective group capable of leaving under reductive conditions or under acidic conditions], and a general formula [IV]: [Wherein X represents a halogen, and a broken line represents a single bond or a double bond], by condensing with cholesterol haloformate or cholestanol haloformate represented by the following general formula [II]: [In the formula, R 1 —O— represents a cholesterol (cholesterol) or cholestanol group bonded at the 3-position. R2 represents a hydroxymethyl, alkyloxycarbonyl, aminocarbonyl, or lower mono- or dialkylaminocarbonyl group. R4 represents a terminal amino group of a lower aminoalkyl group having a protective group capable of leaving under reductive conditions or under acidic conditions], and a basic amino acid derivative represented by the general formula [II ]In
The compound represented by the general formula [I] can be produced by removing the protecting group attached to the terminal amino group of the lower alkyl group as R4.

The compound represented by the general formula [III] is specifically a basic amino acid such as lysine or ornithine, a carboxyl group of which is a reduced form of a hydroxymethyl group, an ester form of which is a t-butoxycarbonyl group, or a compound thereof. Amido-type aminocarbonyl group or 1 to 1 carbon atoms
6 is a linear or compound that was converted to a mono or dialkylamino group consisting of branched alkyl groups, the N ^alpha - Amino groups are free in unsubstituted,
The terminal amino group is a compound having a benzyloxycarbonyl group as a protecting group or a t-butoxycarbonyl group.

Among the basic amino acid derivatives of the compound represented by the general formula [III], a compound in which the R2 moiety has been converted to its reduced form, a hydroxymethyl group, can be obtained by a known method (for example, Shunic
hi Yamada et al . ; Chem. Pharm. 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 [III], an amino acid derivative in which a carboxyl group part is in an ester form or an acid addition salt thereof is used as a starting compound, and a general reduction method is used. For example, by applying a method such as contacting with a reducing agent in a suitable solvent, of the basic amino acid derivative of the compound represented by the general formula [III], the R2 portion is substituted with its reduced form, a hydroxymethyl group. Compound can be obtained.

Examples of the reducing agent in this case include metal hydride compounds such as lithium borohydride, sodium borohydride, and lithium aluminum hydride; and organic metal hydrides such as sodium dihydrobis- (2-methoxyethoxy) aluminate. Examples include a compound or diborane, but are not particularly limited thereto. Further, a halogen compound such as calcium chloride can be used as an additive together with these reducing agents.

As the solvent at the time of reduction, for example, when sodium borohydride is used, solvents such as alcohols and hydrous alcohols can be used, but other solvents that do not affect the reduction reaction, such as dioxane and tetrahydro Ethers such as furan, ethyl ether and diethylene glycol dimethyl ether, or aromatics such as benzene may also be used. As an additive together with these solvents, for example, diethylamine, pyridine, aniline, N
Amines such as -methylamine and γ-collidine may be used in combination.

The reaction temperature during the reduction generally proceeds even at room temperature, but the reaction may be suitably advanced by cooling or heating as appropriate.

Among the basic amino acid derivatives of the compound represented by the general formula [III], a compound in which the R2 moiety has been converted to an alkyloxycarbonyl group which is an ester form thereof, as described above,
Obtain and manufacture the basic amino acid derivative of the compound represented by the general formula [III] in the same manner as in the case of using as a starting material for synthesizing a compound in which the R2 moiety is converted to its reduced form, a hydroxymethyl group. it can.

Among the basic amino acid derivatives of the compound represented by the general formula [III], a compound in which the R2 portion is converted to an ester-type alkyloxycarbonyl group is a carboxyl that is R2 'of the compound represented by the general formula [III]. the moiety conventional method (e.g., J.Am.Chem.Soc 83, 719 (1961) ;. J.Org.Ch
em. , 28 , 1251 (1963), etc.), and an aminocarbonyl group in which R2 is an acid amide type, or a linear or branched chain having 1 to 6 carbon atoms. The compound converted to a mono- or dialkylaminocarbonyl group comprising an alkyl group has the general formula [II
The compound represented by I] can be produced by converting the carboxyl group moiety as R2 'to an acid amide type by a known method.

That is, as a known method, an active esterification method generally used for a peptide bond formation reaction, a mixed acid anhydride method,
Examples include an azide method, an acid chloride method, a symmetric anhydride method, and a method using a coupling reagent. Examples of the coupling reagent include N, N'-cyclohexylcarbodiimide, carbonylimidazole, diphenylphosphoryl azide and the like, and a method using N, N'-cyclohexylcarbodiimide and an additive. In this case, examples of the additive include N-hydroxysuccinimide, 1-hydroxybenzotriazole, N-hydroxy-5-norbornene-, 2,3-dicarboxylic imide and the like.
Examples of the solvent in this case include alcohols such as methanol and ethanol, chloroform, halogenated hydrocarbons such as dichloromethane, tetrahydrofuran,
Examples include ethers such as ether, N, N-dimethylformamide, pyridine and the like, but are not particularly limited thereto.

The compound represented by the general formula [IV] is specifically a haloformate of cholesterol (cholesterol) or cholestanol, and is a known method (for example, H. Wieland).
Chem. 130 , 326- (1923), etc.). It can also be easily obtained as a commercial 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 commonly used.

In order to obtain a compound represented by the general formula [II] by condensing the compounds represented by the general formulas [III] and [IV], both of them can be obtained by a conventional method in an appropriate solvent.
The reaction can be carried out under basic conditions.

As the solvent in this case, for example, alcohols such as methanol and ethanol, chloroform, halogenated hydrocarbons such as dichloromethane, tetrahydrofuran, ether, ethers such as dioxane, benzene, aromatics such as toluene, and further N, N-dimethylformamide,
Examples thereof include pyridine and the like, but are not particularly limited thereto, and any other solvent that does not affect the condensation reaction may be used.

Further, as the base used for the basic condition, for example, trimethylamine, triethylamine,
Examples include tertiary amines such as pyridine, alkali metals such as sodium and potassium, hydroxides, carbonates and hydrogencarbonates of alkaline earth metals such as magnesium and calcium, and particularly, those not limited thereto. Absent.

Further, as for the reaction temperature, this condensation reaction usually proceeds at room temperature, but the reaction may be advantageously promoted by appropriately cooling or heating.

In order to purify the target substance from the reaction station, general recrystallization or column chromatography can be applied.
For example, after completion of the above condensation reaction, the reaction solution is washed with water, the organic phase is extracted, a suitable drying agent (eg, anhydrous sodium sulfate, anhydrous magnesium sulfate, etc.) is added thereto, and after drying, under reduced pressure. The solvent was distilled off, and the residue was subjected to a conventional method.
Recrystallize or use a silica gel column, etc.
Purification can be performed by eluting with an appropriate mixed solvent.

As described above, the compound represented by the general formula [II] can be obtained.
Of the compounds represented by the formula, R2 moiety is an amide-type aminocarbonyl group, or a compound converted to a mono- or dialkylaminocarbonyl group having a linear or branched alkyl group having 1 to 6 carbon atoms, It can also be produced by the following method.

That is, the general formula [III '] [In the formula, R2 ′ represents a carboxyl group. R4 represents a lower aminoalkyl group having a terminal amino group to which a protecting group capable of leaving under reductive conditions has been added under acidic conditions or reductively; and a cholesterol haloformate or halogine represented by the general formula [IV] By condensing with acid cholestanol, the general formula [II '] [In the formula, R 1 —O— represents a cholesterol (cholesterol) or cholestanol group bonded at the 3-position. R2 '
Represents a carboxyl group. R4 represents a group in which a terminal amino group of a lower aminoalkyl group is provided with a protecting group capable of leaving under reductive conditions under acidic conditions or reductively.] To obtain an amino acid derivative represented by the general formula [II ']: R2 'is converted to an alkyloxycarbonyl, aminocarbonyl, or lower mono- or dialkylaminocarbonyl group, and a compound represented by the general formula [II] (where R2 is alkyloxycarbonyl, aminocarbonyl, or lower mono- or Represents only dialkylaminocarbonyl group)
Is a way to get

That is, specifically, it indicates that the carboxyl group of the basic amino acid for protecting the terminal amino group of R4 in the compound represented by the general formula [III '] is free. The compound represented by the general formula [III '] and the compound represented by the general formula [IV] are subjected to a condensation reaction in the same manner as described above to first obtain a compound represented by the general formula [II'], The carboxyl group of the compound represented by [II '] is esterified or amidated by a conventional method as described above, whereby the compound represented by the general formula [II] (where R2 is alkyloxycarbonyl, amino Carbonyl, or only a lower mono- or dialkylaminocarbonyl group).

In this case, for the conditions such as the condensation reaction, esterification or amidation reaction, and the purification step, the above-mentioned conditions for producing the compounds of the general formulas [III] and [II] can be similarly applied (however, in this case, As the base at the time of the condensation reaction, for example, caustic soda or the like can be used more advantageously than triethylamine or the like for the reaction to proceed).

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

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

That is, for example, as a reductive deprotection method, a catalytic reduction method and the like can be mentioned.

Examples of the catalyst used for the catalytic reduction include platinum catalysts such as platinum chloride and platinum oxide, palladium catalysts such as palladium and palladium-carbon, nickel oxide, catalysts such as Raney nickel, and catalysts such as Raney cobalt. Examples of the solvent for the catalytic reduction include water, methanol, ethanol, propanol, N, N-dimethylformamide, acetic acid, formic acid and the like, or a mixed solvent thereof.

Examples of the acid used in the deprotection method under acidic conditions include, for example, trifluoroacetic acid, hydrochloric acid, formic acid, hydrogen fluoride, and the like. Examples of the solvent in this case include dichloromethane, chloroform, dioxane, and tetrahydrofuran. , Alcohols, acetic acid and the like,
It is not particularly limited to these.

The reaction temperature at the time of deprotection usually proceeds even at room temperature, but the reaction can be advantageously advanced by cooling or heating as appropriate.

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

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

The compound represented by the general formula [I] is a natural amino acid derivative obtained by recombining and chemically modifying an amino acid that is a natural product component and a cholesterol derivative, and is an amino acid having high safety and excellent biodegradability. It can be said to be a cationic surfactant.

Further, the compound represented by the general formula [I] is safe and stable because the amino acid and the cholesterol derivative are bonded by urethane, and is advantageous when compounded as a raw material for formulation.

As described in many reports (for example, JP-A-58-134197), amino acid higher alkyl ester salts and their N-lower alkyl-substituted compounds have the following common disadvantages.
It is said that the compound has an ester bond in its structure and therefore has high hydrolyzability, and it is often difficult to stably incorporate it into products.

Furthermore, as a special use, a report using an amino acid derivative as a liposome-forming material (see, for example,
No. 2921). In this report, an amino acid-based anionic surfactant, which is an N-higher acylamino acid, is used as a lipid having a liposome-forming ability, which is easily degraded in a living body and has low toxicity.

In addition, there have been reports of attempts to target liposomes by modifying liposomes with amino acid lipid derivatives (Abstracts of the 7th Meeting of the Biopharmaceutical Society, pp. 49- (1989), etc.). N ^α which is a cationic surfactant
-Research on new properties of liposomes modified with amino acids using cocoyl-arginine ethyl ester (CEA) and the like.

However, the amino acid derivatives used in them all belong to amino acid acyl derivatives, and when applied to living bodies, there is a concern about safety problems such as high hemolysis.

On the other hand, reports on the surface modification of liposomes with cholesterol derivatives having amino groups (for example, KRPate
l et al., Biochimica et Biophysica Acta. 814 , 256-264 .
(1985)), but there is no report on amino acid steroid derivatives as in the present invention.

The constituent parts of liposomes are generally roughly classified into three types: various phospholipids serving as a substrate, cholesterol as a membrane-enhancing substance, and various cationic or anionic surfactants as a membrane-charged substance. In many cases.

In addition, by adding a charged substance to the liposome and imparting a surface charge, aggregation of the liposomes is prevented, an inner aqueous phase is expanded by electrostatic repulsion between the lamellar layers, and a water-soluble encapsulated substance is added to the liposome. It is known that the behavior of a liposome in a living body (for example, tissue transferability) is greatly affected by the type of charged substance used, while increasing the amount of encapsulated material and the like.

When the amino acid-based cationic surfactant of the present invention is used for a liposome membrane material, it has a characteristic that, in terms of its chemical structure, it is both a positively charged substance and cholesterol, a membrane-enhancing substance.

(Effects of the Invention) As is clear from the above description, according to the present invention, as a novel natural amino acid derivative, (1) high safety, excellent biodegradability, amino acid and cholesterol derivative Due to the urethane bond, (2) it can be stably incorporated into a product (formulation). When used as a liposome membrane material, (3) as a safe and novel membrane charged substance, (4) as a membrane charged substance having a structure also serving as a membrane reinforcing 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 has advantages such as the ability to impart unprecedented properties to liposomes.

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

(Examples) Next, the present invention will be described in more detail with reference to Examples, but it should not be construed that the invention is limited thereto.

Reference Example: N ^alpha - Synthesis N ^epsilon amino group protected Rijinoru - benzyloxycarbonyl-lysine methyl ester hydrochloride (Sigma) 3.31 g (10 m mol) in ethanol 10
The mixture was dissolved in 0 ml, and under ice cooling, 2.66 g (70 mmol) of sodium borohydride suspended in 20 ml of ethanol was added dropwise. After the completion of the dropwise addition, 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.
Washed with water. The organic layer was dried over anhydrous magnesium sulfate under reduced pressure, the solvent was distilled off, the resulting crude product was crystallized in acetonitrile, N ^epsilon the desired product - was obtained benzyloxycarbonyl lysine Nord 2.05 g (yield Rate 77%).

Example 1 N ^epsilon - Synthesis chloroformate cholesterol benzyloxycarbonyl lysine Nord cholesterol urethane (Tokyo Kasei Kogyo Co., Ltd.) 2.87 g (6.9
3 mmol) and N ^ε -benzyloxycarbonyl ridinol
Dissolve 1.68 g (6.35 mmol) in 100 ml of dichloromethane,
1.8 ml (12.9 mmol) of triethylamine was added, and the mixture was stirred at room temperature for 6 hours. The reaction solution was washed with water, the organic layer was dried over anhydrous magnesium sulfate, concentrated under reduced pressure to dryness, and the obtained residue was applied to a silica gel column and eluted with a mixed solvent of n-hexane-ethyl acetate (1: 1). The desired fraction was collected from the obtained eluted fraction using thin-layer chromatography (hereinafter abbreviated as TLC), concentrated under reduced pressure to dryness, and then 3.55 g of a solid substance (82% yield).
I got The solid was crystallized in methanol to obtain 3.02 g of N ^ε -benzyloxycarbonylridinol cholesterol urethane as a target product.

Example 2 N ^epsilon - Synthesis chloroformate cholesterol benzyloxycarbonyl lysine Le cholesterol urethane (Tokyo Kasei Kogyo Co., Ltd.) 6.51 g (14.
5 m mol) and N ^epsilon - benzyloxycarbonyl-lysine (Tokyo Chemical Industry Co., Ltd.) 4.09 g (14.6 m mol) was dissolved in chloroform 150 ml, stirring at room temperature, with two portions of 2N sodium hydroxide 14ml this solution 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 dropwise addition of 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 under reduced pressure to dryness, applied to a silica gel column of the obtained residue, and eluted with a mixed solvent of methanol-chloroform (1:20). The obtained target fraction was concentrated under reduced pressure to dryness to obtain 7.43 g (yield 74%) of N ^ε -benzyloxycarbonyl lysine cholesterol urethane solid.

Example 3 N ^epsilon - N was synthesized benzyloxycarbonyl diethylamide lysine cholesterol synthesis in Example 2 of urethane ^epsilon - dissolved benzyloxycarbonyl lysine cholesterol urethane 2.55 g (3.6 mmol) in tetrahydrofuran 50 ml, diethylamine thereto 57
Under ice cooling, 0.94 g (6.1 mmol) of hydroxybenztriazole and 1.51 g (7.3 mmol) of dicyclohexylcarbodiimide were added to 0 μl (5.5 mmol) of the mixture.
After stirring for an hour, the mixture was stirred at room temperature for one day. After the reaction solution was filtered, the filtrate was concentrated under reduced pressure to dryness. 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 sodium hydrogen carbonate, washed twice with water, dried over anhydrous magnesium sulfate, and concentrated to dryness under reduced pressure. The residue was applied to a silica gel column, and eluted with a mixed solvent of n-hexane-ethyl acetate (2: 1). The obtained target fraction was concentrated under reduced pressure to dryness, and N ^ε -benzyloxycarbonyldiethylamidolysine cholesterol urethane solid 2.29 g
(85% yield).

Example 4 Synthesis of lysinol cholesterol urethane 10% palladium carbon (Kojima Chemical Co., Ltd.) (172 mg) was suspended in ethanol (100 ml), and N ^ε -benzyloxycarbonyl ridinol cholesterol urethane 1.
36 g (2.0 mmol) was added, and a normal pressure hydrogenation device (manufactured by Ishii Shokai)
The mixture was stirred at room temperature for 6 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to dryness to obtain 1.083 g (yield: 99%) of a crude product. This crude product was crystallized in ethanol to obtain 0.50 g of the target product, lysinol cholesterol urethane.

 The analysis results are shown below.

R1: Cholesterin No. R2 R3 1. CH ₂ OH (CH ₂ ) ₄ NH ₂ 2. CH ₂ OH (CH ₂ ) ₃ NH ₂ 3. COOt-Bu (CH ₂ ) ₄ NH ₂ 4. C0NH ₂ (CH _{2) 4 NH 2 5. C0NEt 2} (CH 2) 4 NH 2 6. C0NEt 2 (CH 2) 3 physicochemical data mp of NH ₂ compound: (℃) IR: (cm -1) 1 H-NMR: _{(ppm, CDCL 3) MS:} (m / z, M +) No.1 mp: 168~169 IR: 3370,1695,1560,1470,1380,1255 NMR: 0.67 (3H, s), 0.78~2.08 ( 35H, m), 0.83 (6H, d),
0.88 (3H, d), 0.97 (3H, s), 2.13 to 2.42 (2H, m), 2.67
(2H, t), 3.47 ~ 3.72 (3H, m), 4.46 (1H, br), 4.87 (1
H, br), 5.36 (1H, br) MS: 545 No.2 mp: 187-190 IR: 3340,1695,1540,1380,1255 NMR: 0.67 (3H, s), 0.74-2.08 (33H, m) , 0.83 (6H, d),
0.88 (3H, d), 0.97 (3H, s), 2.13 to 2.42 (2H, m), 2.67
(2H, t), 3.47 ~ 3.72 (3H, m), 4.46 (1H, br), 4.87 (1
H, br), 5.36 (1H, br) MS: 531 No.3 mp: 165-168 IR: 3370,1730-1680,1525,1470,1355,1205 NMR: 0.59 (3H, s), 0.72-2.00 ( 34H, m), 0.77 (6H, d),
0.82 (3H, d), 0.92 (3H, s), 1.38 (9H, s), 2.17 ~ 2.32
(2H, m), 2.76 (2H, t), 4.03 (1H, br), 4.36 (1H, br),
5.28 (1H, br) MS: 615 No.4 mp: Amorphous IR: 3405,1705,1660,1610,1540 NMR: 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 (1H, br), 4.47 (1H, br), 5.22 (1H, b
r), 5.36 (1H, br), 5.42 (1H, br), 6.35 (1H, br) MS: 558 No.5 mp: 72-74 IR: 3450,1705,1635,1540,1470,1250 NMR: 0.64 (3H, s), 0.80 ~ 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 to 2.42 (2H, m), 2.70 (2H, t), 3.10 to 3.60 (4
H, m), 4.32 ~ 4.64 (2H, m), 5.35 (1H, br), 5.50 (1H, b
r) MS: 614 No.6 mp: 82-84 IR: 3425,1715,1635,1540,1470,1250 NMR: 0.64 (3H, s), 0.80-2.05 (32H, m), 0.82 (6H, d) ,
0.87 (3H, d), 0.97 (3H, s), 1.08 (3H, t), 1.19 (3H,
t), 2.15 to 2.42 (2H, m), 2.70 (2H, t), 3.10 to 3.62 (4
H, m), 4.32 ~ 4.64 (2H, m), 5.36 (1H, br), 5.50 (1H, b
r) MS: 600 R1: Cholesteryl No. R2 R4 7. CH ₂ OH (CH ₂ ) ₄ NH-CBZ 8. CH ₂ OH (CH ₂ ) ₃ NH-CBZ 9. COOH (CH ₂ ) ₄ NH-CBZ 10. COOH (CH ₂ ) ₄ NH-BOC 11. COOH (CH ₂ ) ₃ NH-CBZ 12. COOH (CH ₂ ) ₃ NH-BOC 13. COOt-Bu (CH ₂ ) ₄ NH-CBZ 14. COOt-Bu (CH ₂ ) ₄ NH-BOC 15. CONH ₂ (CH ₂ ) ₄ NH-CBZ 16. CONH ₂ (CH ₂ ) ₄ NH-BOC 17. CONEt ₂ (CH ₂ ) ₄ NH-CBZ 18. CONEt ₂ (CH ₂ ) ₄ NH- BOC 19. CONEt ₂ (CH ₂ ) ₃ NH-CBZ 20. CONEt ₂ (CH ₂ ) ₃ NH-BOC where CBZ: benzyloxycarbonyl BOC: tert-butoxycarbonyl Physicochemical data mp: (° C) IR ^{: (cm -1) 1 H-} NMR: (ppm) No.7 mp: 161~163 IR: 3440,1710,1695,1540,1260 NMR (CDCL 3): 0.67 (3H, s), 0.78~2.08 ( 33H, m), 0.88
(6H, d), 0.88 (3H, d), 0.97 (3H, s), 2.13-2.42 (2H,
m), 3.19 (2H, br), 3.60 (3H, br), 4.45 (1H, br), 4.73
~ 4.95 (2H, m), 5.06 (2H, s), 5.34 (1H, br), 7.33 (5
H, s) No.8 mp: 130~131 IR: 3440,1695,1540,1265 NMR (CDCL 3): 0.68 (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 (2H, br), 3.62 (3H, br), 4.48 (1H, br), 4.80
~ 4.97 (2H, m), 5.10 (2H, s), 5.37 (1H, br), 7.35 (5
H, s) No.9 mp: amorphous _{IR: 3350,1710~1695,1540,1255 NMR (CDCL 3 +} CD 3 OD): 0.63 (3H, s), 0.74~2.06 (32H,
m), 0.82 (6H, d), 0.87 (3H, d), 0.95 (3H, s), 2.13 ~
2.40 (2H, m), 3.13 (2H, br), 4.18 (1H, br), 4.41 (1H, br
br), 5.04 (2H, s), 5.31 (1H, br), 7.29 (5H, s) No. 10 mp: amorphous IR: 3350, 1720-1695, 1520, 1260 NMR (CDCL ₃ + CD ₃ OD): 0.63 (3H, s), 0.73 ~ 2.05 (32H,
m), 0.83 (6H, d), 0.87 (3H, d), 0.96 (3H, s), 1.38 (9
H, s), 2.12 to 2.38 (2H, m), 3.01 (2H, br), 4.12 (1H, b
r), 4.42 (1H, br), 5.32 (1H, br) No. 11 mp: amorphous IR: 3330, 1715 to 1705, 1540, 1260 NMR (CDCL ₃ + CD ₃ OD): 0.63 (3H, s), 0.74 ~ 2.06 (30H,
m), 0.82 (6H, d), 0.87 (3H, d), 0.95 (3H, s), 2.13 ~
2.40 (2H, m), 3.13 (2H, br), 4.18 (1H, br), 4.41 (1H, br
br), 5.04 (2H, s), 5.32 (1H, br), 7.29 (5H, s) No. 12 mp: amorphous IR: 3350, 1720-1695, 1520, 1250 NMR (CDCL ₃ + CD ₃ OD): 0.63 (3H, s), 0.73 ~ 2.05 (30H,
m), 0.83 (6H, d), 0.87 (3H, d), 0.96 (3H, s), 1.38 (9
H, s), 2.12 to 2.38 (2H, m), 3.01 (2H, br), 4.12 (1H, b
r), 4.42 (1H, br ), 5.32 (1H, br) No.13 mp: amorphous _{IR: 3340,1695,1535,1455,1250 NMR (CDCL 3)} : 0.65 (3H, s), 0.78~2.07 ( 32H, 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 (1H, br), 4.46
(1H, br), 4.79 (1H, br), 5.02 ~ 5.20 (1H, m), 5.07 (2
H, s), 5.34 (1H , br), 7.33 (5H, s) No.14 mp: 140~141 IR: 3370,1720,1705,1525,1370 NMR (CDCL 3): 0.65 (3H, s), 0.80 to 2.05 (32H, m), 0.86
(6H, d), 0.88 (3H, d), 0.98 (3H, s), 1.41 (9H, s), 1.
43 (9H, s), 2.18 to 2.41 (2H, m), 3.08 (2H, br), 4.16
(1H, br), 4.36-4.64 (2H, m), 5.14 (1H, br), 5.34 (1
H, br) No.15 mp: 157~160 IR: 3340,1695,1535,1455,1255 NMR (CDCL 3): 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 (1H, br), 4.49 (1H, br), 4.82
(1H, br), 5.10 (2H, s), 5.26-5.37 (3H, m), 6.21 (1
H, br), 7.33 (5H , s) No.16 mp: 152~155 IR: 3340,1695,1535,1455,1255 NMR (CDCL 3): 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 (2H, br), 4.12 (1H, br), 4.47
(1H, br), 4.63 (1H, br), 5.30-5.43 (3H, m), 5.53 (1
H, br), 6.24 (1H , br) No.17 mp: amorphous _{IR: 3320,1715,1635,1530,1460,1245 NMR (CDCL 3)} : 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 (1H, br), 4.65 (1H, br), 4.91 (1H, br), 5.06 (2
H, s), 5.35 (1H, br), 5.45 (1H, br), 7.33 (5H, s) No.18 mp: Amorphous IR: 3320,1715,1635,1520,1465,1250 NMR (CDCL ₃ ): 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 ~ 4.75 (3H, m), 5.37 (1H, br), 5.47
(1H, s) No.19 mp: 122~125 IR: 3325,1715,1635,1530,1465,1250 NMR (CDCL 3): 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 (1H, br), 4.65 (1H, br), 4.91 (1H, br), 5.06 (1
H, s), 5.35 (1H, br), 5.45 (1H, br), 7.33 (5H, s) No.20 mp: amorphous IR: 3325,1710,1640,1510,1385,1250 NMR (CDCL ₃ ): 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), 3.05-
3.62 (6H, m), 4.34 ~ 4.75 (3H, m), 5.37 (1H, br), 5.47
(1H, s)

────────────────────────────────────────────────── ─── Continuation of the front page (72) Kinya Yamamoto, Inventor 2-7-3, Tachibanadai, Midori-ku, Yokohama-shi, Kanagawa (58) Field surveyed (Int. Cl. ⁶ , DB name) C07J 41/00, 9/00 A61K 9/127

Claims (15)

(57) [Claims] 1. A compound of the general formula [I]: [In the formula, R 1 —O— represents a cholesterol (cholesterol) or cholestanol group bonded at the 3-position. R2 represents a hydroxymethyl, alkyloxycarbonyl, or lower mono- or dialkylaminocarbonyl group. R3 represents a lower aminoalkyl group] and acid addition salts thereof. 2. The general formula [II]: [In the formula, R 1 —O— represents a cholesterol (cholesterol) or cholestanol group bonded at the 3-position. R2 represents a hydroxymethyl, alkyloxycarbonyl, aminocarbonyl, or lower mono- or dialkylaminocarbonyl group. R4 represents a terminal amino group of a lower aminoalkyl group having a protective group capable of leaving under reductive conditions or under acidic conditions]. 3. A basic amino acid derivative according to claim 1, wherein R2 is a hydroxymethyl group, R3 is a 4-aminobutyl or 3-aminopropyl group, and an acid addition salt thereof. 4. The basic amino acid derivative according to claim 1, wherein R2 is a t-butoxycarbonyl group and R3 is a 4-aminobutyl or 3-aminopropyl group, and an acid addition salt thereof. 5. The method according to claim 1, wherein R2 is an aminocarbonyl group, or a linear or branched mono- or dialkylaminocarbonyl group having 1 to 6 carbon atoms;
A basic amino acid derivative which is an aminobutyl or 3-aminopropyl group and an acid addition salt thereof. 6. The method of claim 2, wherein R2 is a hydroxymethyl group, R4 is 4-aminobutyl, or a terminal amino group of a 3-aminopropyl group is provided with a protecting group capable of being removed under acidic conditions or reductively. A basic amino acid derivative. 7. The method according to claim 2, wherein R2 is a t-butoxycarbonyl group, R3 is a 4-aminobutyl, or a terminal amino group of a 3-aminopropyl group under the acidic condition or reductive elimination. A basic amino acid derivative attached. 8. The method according to claim 2, wherein R2 is an aminocarbonyl group, or a mono- or dialkylaminocarbonyl group comprising a linear or branched alkyl having 1 to 6 carbon atoms;
A basic amino acid derivative in which R4 is a 4-aminobutyl or 3-aminopropyl group having a terminal amino group with a protecting group capable of leaving under reducing or acidic conditions. 9. The basic amino acid derivative according to claim 2, wherein the protecting group for the terminal amino group of the lower aminoalkyl group as R4 is a benzyloxycarbonyl group or a t-butoxycarbonyl group. 10. The general formula [III]: [In the formula, R2 represents a hydroxymethyl, alkyloxycarbonyl, aminocarbonyl, or lower mono- or dialkylaminocarbonyl group. R4 represents a terminal amino group of a lower aminoalkyl group having a protective group capable of leaving under reductive conditions or under acidic conditions], and a general formula [IV]: [Wherein X represents a halogen, and a broken line represents a single bond or a double bond], by condensing with cholesterol haloformate or cholestanol haloformate represented by the following general formula [II]: [In the formula, R 1 —O— represents a cholesterol (cholesterol) or cholestanol group bonded at the 3-position. R2 represents a hydroxymethyl, alkyloxycarbonyl, aminocarbonyl, or lower mono- or dialkylaminocarbonyl group. R4 represents a terminal amino group of a lower aminoalkyl group having a protecting group capable of leaving under reductive conditions or under acidic conditions], to produce a compound represented by the formula: Method. [11] a protecting group attached to a terminal amino group of a lower alkyl group as R4 in the compound represented by the general formula [II] produced in the above [10], I]: [In the formula, R 1 —O— represents a cholesterol (cholesterol) or cholestanol group bonded at the 3-position. R2 represents a hydroxymethyl, alkyloxycarbonyl, or lower mono- or dialkylaminocarbonyl group. R3 represents a lower aminoalkyl group]. The method for producing a compound according to claim 1, wherein 12. A compound of the general formula [III ']: [Wherein R 2 ′ represents a carboxyl group, R 4 represents a lower aminoalkyl group having a terminal amino group to which a protective group capable of leaving under reductive elimination under acidic conditions] and a general formula [ By condensing with cholesterol haloformate or cholestanol haloformate represented by the formula [II '] [In the formula, R1 represents a cholesterol (cholesterol) or cholestanol group bonded at the 3-position. R2 'represents a carboxyl group. R4 represents a lower aminoalkyl group having a terminal amino group having a protective group capable of leaving under reductive conditions under acidic conditions or reductive conditions], and then obtaining an amino acid derivative represented by the general formula [II ']. The carboxyl group of R2 'in the compound to be converted is converted to an alkyloxycarbonyl, aminocarbonyl or lower mono- or dialkylaminocarbonyl group to obtain a compound represented by the general formula [II] (in this case, alkyloxycarbonyl, aminocarbonyl, or 3. The process for producing a compound according to claim 2, which comprises producing a lower mono- or dialkylcarbonyl group only). 13. The compound represented by the general formula [I], wherein the protecting group attached to the terminal amino group of the lower alkyl group as R4 in the compound represented by the general formula [II] produced in claim 12 is eliminated. 2. A method for producing a compound according to claim 1, wherein the compound is produced by the method described above, wherein R2 represents only an alkyloxycarbonyl, aminocarbonyl, or lower mono- or dialkylaminocarbonyl group. 14. A compound represented by the formula (I) or (II):
By hydrogenation to a compound representing a cholesterol (cholesterol) group in which O- is bonded at the 3-position, R1-O- in the general formula [I] or [II] is bonded at the 3-position 3. The method for producing a compound according to claim 1, wherein the compound representing a cholestanol group is produced. 15. The method according to claim 10, wherein the compound represented by the general formula [I
V], wherein the halogen of X is chlorine or bromine.