JPH0680560A - Production of liposome - Google Patents

Abstract

PURPOSE:To provide a liposome having excellent stability, reticuloendothelial avoidance, organotropic property, drugretaining function, etc. CONSTITUTION:The objective liposome can be prepared by compounding (i) a polar lipid, (ii) a compound giving positive or negative charge, (iii) cholesterol, (iv) a compound containing a polyethylene glycol unit having a polymerization degree of 3-6 and at least two 5-20C alkyl groups (or its lipid derivative), (v) a drug component and (vi) a solvent at specific ratios.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for producing a drug-holding liposome as a so-called drug delivery system, and more specifically to a method for producing a liposome which facilitates carrying an organ-oriented sensor.

[0002]

2. Description of the Related Art Liposomes and lipid microspheres are used as one means of drug delivery system for delivering effective drug treatment by delivering a drug to be administered to a living body to a required tissue in a required amount at a required time. It is known to utilize particulate carriers such as. However, when these fine particle carriers are administered intravascularly, they are easily trapped in the reticuloendothelial tissues represented by the liver, spleen, etc., and therefore, sustained-release preparations that control drug release and drugs for target tissues. It is well known that there are still problems in using targeting formulations for delivery.

Therefore, in order to solve this problem, it has been attempted to make the liposome carry an organ-directing sensor for sugars, antibodies and the like. That is, the liposome is coated with the lipid derivative of the organ-directed sensor to expose the organ-directed sensor on the surface layer of the liposome. However, according to this conventional method, in some cases, the organ directional sensor does not correctly or frequently recognize the organ, and in some cases,
There is a problem that the lipid derivative of the organ-directed sensor does not coat the liposome efficiently, and in some cases, the particle size of the liposome is not constant, and in other cases, the stability of the liposome is low. It was

[0004]

DISCLOSURE OF THE INVENTION The present invention aims to provide a liposome which is excellent in stability, avoidance of microcapillary endothelium tissue, excellent in directing to a specific organ, and excellent in drug retention function. To aim.

[0005]

The present inventor has accomplished the present invention as a result of earnest research to solve the above problems.

The present invention will be sequentially described below.

The present invention provides, as a raw material, at least 1 mol of a polar lipid, a compound 0.05 which gives a positive or negative charge.
.About.0.5 mol, cholesterol 0.3 to 1.5 mol, polyethylene glycol having a degree of polymerization of 3 to 6 and a compound having at least two alkyl groups having 5 to 20 carbon atoms in the molecule 0.02
.About.0.5 mol and 50 to 100 l of aqueous solvent in this proportion are used.

[0008] Polar lipids are the main constituents that form the membrane of liposomes. Examples of such polar lipids include polar lipids such as diphosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine, and phosphatidylcholine, and mixtures of these polar lipids, and phospholipids such as phosphatidylcholine are particularly preferable. As the phosphatidylcholine, dimyristoylphosphatidylcholine, dipalmitoylphosphatidylcholine, distearoylphosphatidylcholine, egg yolk lecithin, soybean lecithin and the like are preferable.

Examples of the compound which gives a positive charge include:
Aliphatic amines such as stearylamine, and examples of compounds that impart a negative charge include dicetyl phosphoric acid. These compounds are 0.05 mols per mol of polar lipid.
Used in proportions of ~ 0.5 mol.

Cholesterol is used as a stabilizer, and if the amount used is too large, the formation of liposomes is inhibited, or even if liposomes are formed, it is physicochemically unstable. Can not form stable liposomes. Therefore, 0.3 to 1.5 per mole of polar lipid
Preference is given to use in molar proportions.

A compound having polyethylene glycol having a degree of polymerization of 3 to 6 and at least two alkyl groups having 5 to 20 carbon atoms in the molecule (hereinafter, may be referred to as "lipid derivative") is coated on a liposome to form an organ. It serves as a directional sensor.

Polyethylene glycol having a degree of polymerization of ethylene glycol of 3 to 6 must be used. Even if a polymerization degree other than the above is used, the desired result cannot be obtained.

Explaining the alkyl group having 5 to 20 carbon atoms, which should be contained in at least two in the molecule of the lipid derivative, polyethylene glycol is a functional group capable of directly bonding to the hydroxyl group in the lipid derivative molecule. There are cases where an alkyl group having a carboxyl group is bonded and cases where an alkyl group is bonded to a linker having a functional group capable of bonding to the hydroxyl group, such as glutamic acid. The latter is advantageous because it allows two alkyl groups to be attached to the linker.

As the donor of the alkyl group, a fatty acid having 5 to 20 carbon atoms can be mentioned as an example of the former case.
And as an example of the latter case, a C5-C20 fatty acid, an aliphatic alcohol, and an aliphatic primary amine can be mentioned. These fatty acids, fatty alcohols, fatty amines and the like may be either linear or branched.
Also, when the hydroxyl group of polyethylene glycol is replaced with an amino group or other reactive functional group, as in the above case, when the alkyl group is directly bonded to the reactive functional group and the linker is In some cases, they are linked via. Examples of the alkyl group donor include the above-mentioned fatty acids having 5 to 20 carbon atoms, aliphatic alcohols and aliphatic amines.

At least 2 alkyl groups having 5 to 20 carbon atoms
The alkyl group when it has one may be one in which two or more alkyl groups are connected via a linker such as ethylene glycol or glutamic acid, but each alkyl group has 5 to 20 carbon atoms.

The number of carbon atoms of the alkyl group to be contained in the lipid derivative of the present invention is 5 to 20, preferably 12 to 18 in both straight chain and branched chain alkyl groups, but depending on the type of linker, Even if the number of carbon atoms is 6, preferable results can be obtained.

In the lipid derivative, the carbon atom at one end of the alkyl group has a functional group capable of binding to polyethylene glycol or a linker as described above, but the carbon atom at the other end is not bound to anything other than a hydrogen atom. It is a methyl group (that is, it is not functionalized).

As described above, the hydroxyl group at one end of the polyethylene glycol of the lipid derivative is bonded to the functional group of the alkyl group directly or through a linker by itself or by substituting with another functional group such as an amino group. , The other end is an antibody,
Ligands such as antigens, lectins and cell adhesion factors; proteins such as cell adhesion factors, antibodies and immunoglobulins; saccharides; amino acids; nucleic acids and nucleic acid compounds; and are responsible for organ-oriented sensors such as compounds having various polarities .

The ratio of the lipid derivative to the polar lipid has a great influence on the quality of the obtained liposome. That is,
If it is too large, stable liposomes cannot be produced, and if it is too small, it does not change the liposome membrane to be obtained, and the effect of incorporating the lipid derivative is not exhibited. For this reason, the lipid derivative is used for 1 mol of polar lipid.
It is used in a proportion of 0.02 to 0.5 mol, preferably 0.1 to 0.5 mol.

Drugs to be contained in the liposome include various therapeutic agents such as anticancer agents and antifungal agents, as well as test agents. These include proteins, saccharides, nucleic acids and nucleic acid-based compounds, peptides and various synthetic compounds.
These agents are usually used by dissolving them in an aqueous solvent, but they may be used by adding them to an organic solvent.

The concentrations of these drugs in the aqueous solvent are not particularly limited and can be appropriately determined depending on the method of use and the application of the liposome. The aqueous solvent is preferably an isotonic solution such as physiological saline, and the pH must not be extremely acidic or alkaline.

When the drug is added to the organic solvent, the drug is not added to the aqueous solvent, but the method for preparing liposomes is the same as when the drug is added to the aqueous solvent.

An aqueous solution of the drug is used for 1 mol of polar lipid.
Used at a rate of 50-100 liters.

The preparation method itself for preparing drug-loaded liposomes using polar lipids, compounds imparting positive or negative charges, cholesterol, lipid derivatives and aqueous solutions of drugs as raw materials in the above-mentioned proportions is a special method. It doesn't have to be. For example, Nojima et al. “Liposome” (Nankodo 1989
A large number of methods for preparing liposomes are listed in (September 15, 2013), and any of these methods can be used in the present invention. The method of Bangham as a method for preparing multilamellar liposomes (AD Bangham et al., J. Mol. Biol., 13 ,
238 (1965) is known, in this method, the solvent is removed from the organic solvent solution of the raw material of the liposome by nitrogen gas flow or vacuum distillation to form a lipid film on the container wall, and an aqueous solvent is added to this. In addition, the lipid film is allowed to stand for a while to hydrate and then exposed to ultrasonic waves or vortexed to prepare liposomes. The resulting multilamellar liposomes have a large particle size, and the encapsulated substance (drug)
Also has a high retention capacity. On the other hand, small unilamellar vesicles can be prepared by an ultrasonic treatment method, an ethanol injection method, a French press method, etc., but generally have a low retention volume for a water-soluble drug. However, depending on the type of drug, good results may be obtained. On the other hand, ether injection method, cholic acid (surfactant) method, calcium ion fusion method, freezing-
The method for preparing large unilamellar vesicles such as the melting method and the reverse phase evaporation method is preferable because relatively stable liposomes can be obtained and the amount of the water-soluble drug retained is large.

The particle size of the drug-carrying liposome of the present invention is 50 to 1000 nm, more preferably 50 to 300 nm, from the viewpoint of stability and pharmacokinetics. The particle size of the liposome can be adjusted by, for example, pressure filtration. When it is necessary to select liposomes having a particle size within the above range, a usual method such as a gel filtration method or a membrane separation method may be appropriately improved and used.

The following three are exemplified as more preferable embodiments of the present invention. The first is 1 mol of a polar lipid, 0.05 to 0.5 mol of a compound that gives a positive or negative charge, 0.3 to 1.5 mol of cholesterol and polyethylene glycol having a degree of polymerization of 3 to 6 in a molecule, and at least two of 5 to 20 carbon atoms.
The compound containing 0.02 to 0.5 mol of the compound having an alkyl group is removed from the organic solvent to form a lipid film, and then an aqueous solution of the drug to be incorporated into the liposome is added in an amount of 50 to 100 per 1 mol of the phospholipid. 1 is added to the lipid film to form liposomes, and the particle size is 50
The liposomes of ~ 1000 nm are selectively collected.

There is no particular limitation on the organic solvent, but it is preferable that the organic solvent dissolves polar lipids, cholesterol and lipid derivatives and can be easily removed, for example, chloroform, ether,
A chloroform-methanol mixed solution and the like can be mentioned.

The concentration of the organic solvent solution is not particularly limited as long as the organic solvent can dissolve the polar lipid.

The lipid film can be prepared from the organic solvent solution by, for example, removing the organic solvent with a nitrogen gas stream to form a lipid film on the container wall, or distilling the organic solvent off under reduced pressure.

There is no particular limitation on the method for preparing a drug-supporting liposome by adding an aqueous solution of a drug to a lipid film. For example, the aqueous solution is added to the lipid film and left standing for a while to hydrate the lipid film. It can be followed by exposure to ultrasound or vortexing.

From the liposomes thus prepared, those having a particle size of 50 to 1000 nm are selectively collected. The selection method itself can be a usual method such as a gel filtration method or a molecular sieve membrane method.

In a second embodiment, 1 mol of polar lipid, 0.05 to 0.5 mol of a compound that gives a positive or negative charge, 0.3 to 1.5 mol of cholesterol and polyethylene glycol having a degree of polymerization of 3 to 6 and at least two are included in the molecule. A compound having an alkyl group having 5 to 20 carbon atoms, 0.02 to 0.
An aqueous solution of the drug to be incorporated into the liposome in an organic solvent containing 5 mol in this ratio is 50 to 50 mol per mol of the phospholipid.
100 l was added, then the mixed solution was sonicated and w
/ O type emulsion, the organic solvent is removed under reduced pressure, vortexing is further performed to form an o / w type emulsion, and then the organic solvent is removed again under reduced pressure, if necessary, by the so-called reverse evaporation method.

The purpose of subjecting the mixed solution of the organic solvent solution and the aqueous solution of the drug to ultrasonic treatment is to prepare a w / o emulsion, and therefore, this ultrasonic treatment is specifically carried out under known ordinary conditions. be able to. With respect to the removal of the organic solvent, it is not necessary to change the method to a usual method in the present invention.

Vortexing is for the purpose of converting a w / o emulsion into an o / w emulsion. When the organic solvent is removed from the obtained o / w emulsion under reduced pressure, the target drug-supporting liposome is obtained.

The third embodiment uses the drug as an organic solvent solution, whereas the above two embodiments use the drug as an aqueous solvent solution.

The drug-carrying liposome prepared by the method of the present invention, that is, a liposome containing a lipid derivative having at least two alkyl groups is the same as the method of the present invention except that a compound having one alkyl group is used. Compared with liposomes produced by the same method, when an organ-directing sensor is carried at one end of ethylene glycol, the organ recognition property of the liposomes is improved, and the accumulation property of the liposomes in the organs or tissues is increased.

[0037]

【Example】

Example 1 (Synthesis Example (1)) 4 having polyethylene glycol having a degree of polymerization of 3 to 6 and at least two alkyl groups having 5 to 20 carbon atoms in a molecule
The compounds of the present invention (invention) 256, 852, 331 and 531 were each synthesized as follows.

The reactions in these synthetic reactions are shown in FIGS.
Shown in.

(A) Synthesis of compound 256 (FIG. 1) 1) Synthesis of compound 254 Galactose peracetate (10.0 g, 25.62 mmol) and monochlorotriethylene glycol (5.616 g, 33.3 mm)
To a methylene chloride (150 ml) solution of ol, 1.3 eq), a solution of boron trifluoride ether complex (12.6 ml, 4.0 eq) in methylene chloride (30 ml) was added under ice cooling, and the mixture was stirred overnight at room temperature. The resulting solution was added to ice water and extracted with chloroform (150 ml). The organic tank was washed twice with water and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure,
The residue was separated by 1000 ml silica gel column chromatography (hexane: ethyl acetate = 2: 1-1: 1),
The desired product was obtained (6.51 g, 50.9%).

[Α] _D ²⁰ = -0.4 ° (c 1.098, chloroform).

¹ H-NMR (CDCl ₃ , δ); 1.986, 2.051,
2.063, 2.152 (4s, 3H x 4), 3.63-3.78 (m, 11H), 3.95-
3.98 (m, 1H), 3.917 (brt, 1H), 4.131 (dd, 1H, J = 6.8Hz, 1
1.2Hz), 4.178 (dd, 1H, J = 6.6Hz), 4.576 (d, 1H, J = 8.1Hz)
, 5.023 (dd, 1H, J = 3.4Hz), 5.212 (dd, 1H, J = 10.5Hz),
5.390 (brd, 1H).

2) Synthesis of Compound 225 Compound 254 (3.445 g, 12.91 mmol) and sodium azide (1.26 g, 19.4 mmol, 1.5 eq) in DMF (50 ml).
Was added, and the mixture was heated with stirring at 60 ° C. for 17 hours. Water (100 ml) was added to the obtained solution, and the mixture was extracted with ethyl acetate. The organic layer was washed with water and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was separated by 500 ml silica gel column chromatography (hexane: ethyl acetate = 2: 1.5).
-1: 1) The desired product was obtained (5.30 g, 81.2%).

[Α] _D ¹⁷ = -3.2 ° (c 1.04, CHCl ₃ ).

¹ H-NMR (CDCl ₃ , δ); 1.984, 2.048,
2.060, 2.148 (4s, 3H x 4), 3.398 (t, 2H, J = 5.0Hz), 3.
63-3.69 (m, 8H), 3.73-3.78 (m, 1H), 3.95-3.98 (m, 1H)
， 3.910 (brt, 1H) ， 4.131 (dd, 1H, J = 6.8Hz, 11.2Hz) ，
4.176 (dd, 1H, J = 6.3Hz), 4.571 (d, 1H, J = 7.8Hz), 5.023
(dd, 1H, J = 3.4Hz), 5.210 (dd, 1H, J = 10.5Hz), 5.387 (dd,
1H, J = 1.0Hz).

3) Compound 225 (0.417 g) and paratolusulfonic acid monohydrate (0.190 g) in methanol (20 ml)
And ethyl acetate (40 ml) were added and dissolved. Lindlar's catalyst (0.2 g) was added to the solution, and the mixture was stirred under a hydrogen atmosphere of 50 psi for 8 hours. The catalyst was filtered off, and the solvent was distilled off under reduced pressure. The residue is methylene chloride (20 ml) and hexane (10 ml)
Dissolved in 2-palmitylstearic acid (0.662 g),
N-hydroxysuccinic acid (0.150 g), triethylamine (0.181 ml) and dicyclohexylcarbodiimide
(0.268 g) was added, and the mixture was stirred overnight. The solution was concentrated under reduced pressure, ethyl acetate was added to the residue, and the insoluble material was filtered off. The filtrate was concentrated and separated by silica gel column chromatography (hexane: ethyl acetate = 2: 1) to obtain the desired product (0.504 g, 57.1%).

[Α] ^D ₁₈ = -0.4 ° (c 1.04, chloroform).

¹ H-NMR (CDCl ₃ , δ); 0.88 (t, 6H, J = 7.0H
z), 1.18-1.26 (m, 56H), 1.34-1.43 (m, 2H), 1.52-1.62
(m, 2H), 1.99 (bs, 4H), 2.05 (s, 3H), 2.06 (s, 3H), 2.1
5 (s, 3H), 3.46-3.49 (m, 2H), 3.54 (t, 2H, J = 5.0Hz), 3.5
9-3.67 (m, 6H), 3.74 (ddd, 1H, J = 3.6Hz, 7.2Hz, 10.8Hz)
, 3.90-3.93 (m, 1H), 3.98 (dt, 1H, J = 4.4Hz), 4.13 (d
d, 1H, J = 6.8Hz, 11.2Hz), 4.18 (dd, 1H, J = 6.6Hz, 11.2Hz),
4.55 (d, 1H, J = 7.9Hz), 5.02 (dd, 1H, J = 3.4Hz, 10.5Hz), 5.
21 (dd, 1H, J = 7.9Hz, 10.5Hz), 5.39 (dd, 1H, J = 3.4Hz, 1.0H
z), 5.92 (t, 1H, J = 5.6Hz).

4) Synthesis of compound 256 Compound 225 (341 mg) was dissolved in benzene (10 ml) to give 5
Eight drops of M / 1 sodium methylate methanol solution were added and stirred overnight. Strong acidic ion exchange resin "Do"
Wex 50w × 8 ”H type was added to neutralize. The solution was filtered, concentrated under reduced pressure, and the residue was treated with "Sephadex LH-2.
0 ”(chloroform: methanol = 9: 1, 22mmφ × 4
5cm) to obtain the target compound (252mg, 90
%).

[Α] ^D ₂₀ = -1.1 ° (c 1.12, chloroform: methanol = 9: 1).

¹ H-NMR (pyridine-d ₅ -D ₂ O, δ); 0.88 (t,
6H, J = 7.0Hz), 1.21-1.39 (m, 52H), 1.43-1.62 (m, 6H),
1.91-1.99 (m, 6H), 2.51-2.57 (m, 1H), 3.61-3.66 (4H,
m), 3.70-3.78 (6H, m), 3.93 (dt, 1H, J = 5.3Hz, 10.7H
z), 4.02-4.04 (m, 1H), 4.13 (dd, J = 3.3Hz, 9.4Hz), 4.26
(dt, 1H, J = 10.7Hz, 4.8Hz), 4.40-4.44 (m, 3H), 4.54 (bd,
1H), 4.78 (d, 1H, J = 7.8Hz), 8.76 (bt, 1H).

(B) Synthesis of Compound 852 (FIG. 2) Carboxylic acid 828 (180 mg) and N-hydroxysuccinimide (41 mg) in methylene chloride solution (10 ml) were added with N,
N'-Dicyclohexylcarbodiimide (74 mg) was added, and the mixture was stirred at room temperature for 1 hr. A solution of the para-toluenesulfonate of the amino compound 818 (190 mg) and triethylamine (90 μl) in methylene chloride (5 ml) was added to this solution, and the mixture was stirred at room temperature overnight. After the insoluble matter was filtered off, the filtrate was washed with water and half-saturated saline, and dried over magnesium sulfate. The solvent was distilled off, and the residue was purified by silica gel column chromatography (silica gel 40 g, chloroform: methanol = 98: 2) to give an amide compound (310 mg, 77).
%) Was obtained.

IR (KBr): 3700, 3600, 1745, 1710, 151
0, 1480, 1420 cm ^-1 .

¹ H-NMR (CD ₃ OD): 0.90 (3H, t, J = 7Hz), 1.9
3 (3H, s), 1.95 (3H, s), 2.03 (3H, s), 2.14 (3H, s), 4.65
(1H, d, J = 8.5Hz), 5.07 (1H, dd, J = 3.5,11.5Hz), 5.33 (1H,
d, J = 3.5 Hz).

R _f = 0.6 (chloroform: methanol = 93: 7).

Sodium methoxide (55 μl of 28% methanol solution) was added to a solution of the amide compound (270 mg) obtained above in methanol (20 ml), and the mixture was stirred at room temperature for 5.5 hours.
"Amberlyst 15E" was added until the liquid became neutral, the resin was filtered off, and the filtrate was concentrated to give the desired compound 852 (21
5 mg) was obtained.

[Α] _D ²⁶ + 43.9 ° (c 1.0, methanol).

IR (KBr): 3340, 3330, 1655, 1555, 1470 cm ^-1 .

¹ H-NMR (CD ₃ OD): 0.90 (3H, t, J = 7Hz), 2.0
0 (3H, s), 3.84 (1H, d, J = 3Hz), 4.45 (1H, dJ = 8.3H).

R _f = 0.37 (chloroform: methanol = 9: 1).

(C) Synthesis of Compound 331 (FIG. 3) 1) Synthesis of Compound 327 α-D-mannose pentaacetate (Compound 302)
(3.90 g) and 2- [2- (2-chloroethoxy) ethoxy] ethanol (3.37 g) were dissolved in methylene chloride (200 ml), BF ₃ · Et ₂ O (5.68 g) was added, and the mixture was stirred at room temperature for 5 hours.
It was stirred for a day. Dilute the reaction solution with methylene chloride, and add water, 5
% NaHCO ₃ water and water, and the solvent was distilled off under reduced pressure. The residue was purified by column chromatography (chloroform) using silica gel (200 g), α
-Glycoside compound 327 (4.13g) was obtained as a colorless oil.

[Α] _D + 38.7 ° (c 1.51, CHCl ₃ ).

¹ H-NMR (CDCl ₃ ) δ: 2.04 (3H, s), 2.05 (3
H, s), 2.10 (3H, s), 2.16 (3H, s), 3.63-3.70 (9H, m), 3.
75-3.85 (3H, m), 4.07 (1H, ddd, J = 2.4Hz, 5.1Hz, 10.0Hz)
, 4.11 (1H, dd, J = 2.4Hz, 12.2Hz), 4.29 (1H, dd, J = 4.9Hz,
12.2Hz), 4.88 (1H, d, J = 1.5Hz), 5.27 (1H, dd, J = 1.5Hz, 3.
4Hz), 5.29 (1H, t, J = 10.0Hz), 5.36 (1H, dd, J = 3.4Hz, 1
0.0Hz).

2) Synthesis of Compound 328 Chloro compound 327 (3.68 g) was dissolved in DMF (50 ml), sodium azide (0.72 g) was added, and the mixture was stirred at 60 ° C. for 24 hours. The reaction solution was diluted with ethyl acetate, washed with water, dried and the solvent was evaporated under reduced pressure. The residue is silica gel (150 g)
Column chromatography (chloroform-acetone 10: 1) using azide compound 328 (3.05
g) was obtained as a colorless oil.

[Α] _D + 35.6 ° (c 2.54, CHCl ₃ ).

¹ H-NMR (CDCl ₃ ) δ: 1.99 (3H, s), 2.04 (3
H, s), 2.11 (3H, s), 2.16 (3H, s), 3.40 (2H, t, J = 5.1Hz),
3.62-3.86 (10H, m), 4.06 (1H, ddd, J = 2.4Hz, 4.9Hz, 10.0H
z), 4.10 (1H, dd, J = 2.4Hz, 12.2Hz), 4.29 (1H, dd, J = 4.9H
z, 12.2Hz), 4.88 (1H, d, J = 1.5Hz), 5.27 (1H, dd, J = 1.5Hz,
3.4Hz), 5.29 (1H, t, J = 10.0Hz), 5.36 (1H, dd, J = 3.4Hz,
10.0 Hz).

3) Synthesis of compound 329 Azide compound 328 (386 mg) and p-toluenesulfonic acid (145 mg) were dissolved in ethanol (20 ml), Lindlar catalyst (770 mg) was added, and catalytic reduction was carried out at room temperature of 50 psi for 7.5 hours. went. After removing the catalyst by filtration, the filtrate was concentrated under reduced pressure to obtain an amine compound 329.

4) Synthesis of compound 330 2- (1-hexadecyl) octadecanoic acid (153 mg) was dissolved in a mixed solvent of hexane (15 ml) and methylene chloride (20 ml), and N-hydroxysuccinimide (35 mg) and N,
N'-Zinclohexylcarbodiimide (62 mg) was added, and the mixture was stirred at room temperature for 24 hours. The amine compound 329 (235 mg) dissolved in acetonitrile (15 ml) was added to the reaction mixture, and then toluethylamine (67 mg) was added, followed by stirring at room temperature for 23 hours. The insoluble matter was filtered off, the filtrate was diluted with chloroform, washed with water, dried and the solvent was distilled off under reduced pressure.
The residue was purified by column chromatography (chloroform-methanol 150: 1) using silica gel (60 g). The product was purified again by column chromatography using silica gel (60 g) (hexane-ethyl acetate 5: 4) to obtain the target compound (130 mg) as a colorless oil.

[Α] _D + 18.2 ° (c 1.02, CHCl ₃ ).

¹ H-NMR (CDCl ₃ ) δ; 0.88 (6H, t, J = 6.8H
z), 1.20-1.33 (56H, m), 1.34-1.43 (2H, m), 1.53-1.62
(2H, m), 2.00 (3H, s), 2.04 (3H, s), 2.11 (3H, s), 2.16
(3H, s), 3.47 (2H, q), 3.55 (2H, t), 3.60-3.73 (7H, m),
3.80-3.85 (1H, m), 4.07 (1H, ddd, J = 2.4Hz, 4.9Hz, 10.0H
z), 4.12 (1H, dd, J = 2.4Hz, 12.2Hz), 4.29 (1H, dd, J = 4.9H
z), 4.89 (1H, d, J = 1.7Hz), 5.27 (1H, dd, J = 1.7Hz, 3.4Hz)
, 5.30 (1H, t, J = 10.0Hz), 5.36 (1H, dd, J = 3.4Hz, 10.0H
z), 6.00 (1H, m).

5) Synthesis of Compound 331 Compound 330 (123 mg) was dissolved in methanol (10 ml),
28% NaOME in MeOH (20 μl) was added, and the mixture was stirred at room temperature for 4 hours. "Amberlite IRC-
50 "was added, the insoluble material was filtered off, and the filtrate was concentrated to dryness under reduced pressure. Chloroform was added to the residue, the insoluble material was filtered off, the filtrate was concentrated to dryness under reduced pressure, and the residue was washed with hexane under ice-cooling to obtain the target compound 231 (101 mg) as a colorless powder.

[Α] _D + 19.5 ° (c 0.59, CHCl ₃ ).

¹ H-NMR (CDCl ₃ ) δ: 0.88 (6H, t, J = 6.8H
z), 1.20-1.33 (56H, m), 1.34-1.44 (2H, m), 1.52-1.61
(2H, m), 2.00 (1H, m), 3.42-3.96 (18H, m), 4.89 (1H,
s (, 6.10 (1H, m).

(D) Synthesis of Compound 531 (FIG. 4) 1) Synthesis of Compound 530 2- (1-hexadecyl) octadecanoic acid (153 mg) was dissolved in a mixed solution of hexane (15 ml) and methylene chloride (20 ml), N-hydroxysuccinimide (35 mg) and N,
N'-Dicyclohexylcarbodiimide (62 mg) was added, and the mixture was stirred at room temperature for 24 hours. Amine compound 527 (214 mg) in which acetonitrile (15 ml) was dissolved was added to the reaction mixture, and then triethylamine (67 mg) was added, followed by stirring at room temperature for 24 hours. The insoluble matter was filtered off, the filtrate was diluted with chloroform, washed with water, dried and the solvent was distilled off under reduced pressure.
The residue was purified by column chromatography (chloroform-methanol 150: 1) using silica gel (60 g). It was again purified by column chromatography using silica gel (60 g) (hexane-ethyl acetate 2: 1) to obtain the target compound 530 (165 mg) as a colorless oil.

[Α] _D -51.4 ° (c 0.84, CHCl ₃ ).

¹ H-NMR (CDCl ₃ ) δ: 0.88 (6H, t, J = 6.8H
z), 1.14 (3H, d, J = 6.4Hz), 1.20-1.33 (56H, m), 1.34-1.4
3 (2H, m), 1.56-1.62 (2H, m), 1.99 (3H, s), 2.07 (3H,
s), 2.17 (3H, s), 3.47 (2H, m), 3.54 (2H, m), 3.58-3.70
(7H, m), 3.76-3.83 (1H, m), 4.22 (1H, dq, J = 1.2Hz, 6.4
Hz), 5.12 (1H, dd, J = 3.7Hz, 10.0Hz), 5.13 (1H, d, J = 3.7H)
z), 5.29 (1H, dd, J = 1.2Hz, 3.4Hz), 5.37 (1H, dd, J = 3.4Hz, 1
0.0Hz), 6.02 (1H, m).

2) Synthesis of Compound 531 Compound 530 (138 mg) was dissolved in methanol (10 ml),
28% NaOMe in MeOH (20 μl) was added, and the mixture was stirred at room temperature for 2.5 hours. "Amberlite IRC"
-50 "was added, the insoluble material was filtered off, and the filtrate was concentrated to dryness under reduced pressure. Chloroform was added to the residue, the insoluble material was filtered off, the filtrate was concentrated to dryness under reduced pressure, and the residue was washed with ether to obtain the target compound (101 mg) as a colorless powder.

[Α] _D -36.7 ° (c 0.69, CHCl ₃ ).

¹ H-NMR (CDCl ₃ ) δ: 0.88 (6H, t, J = 6.8H
z), 1.20-1.30 (56H, m), 1.30 (3H, d, J = 6.8Hz), 1.34-1.4
3 (2H, m), 3.37-3.72 (11H, m), 3.74-3.85 (3H, m), 3.88
-3.94 (1H, m), 4.03 (1H, q, J = 6.8Hz), 4.92 (1H, d, J = 3.4H
z), 6.37 (1H, m).

As controls, the above compounds 256, 852, 331
And 431 each having one alkyl group having 5 to 20 carbon atoms (control compound) 22
8, 851, 333 and 529 were synthesized as follows.

The reactions in these synthetic reactions are shown in FIGS.
Shown in.

(A ′) Synthesis of compound 228 (FIG. 5) 1) Synthesis of compound 225 β-D-galactose pentaacetate compounds 201, 5.
254 g and 3.066 g of 2- [2- (2-azidoethoxy) ethoxy] ethanol were dissolved in 50 ml of methylene chloride,
The mixture was stirred under ice cooling. 6.62 ml of boron trifluoride diethyl ether complex was dissolved in 10 ml of methylene chloride and added dropwise over 10 minutes. After stirring at room temperature for 14 hours, the mixture was poured into ice water and the organic layer was separated. After washing three times with water (the aqueous layer became neutral), the mixture was washed with saturated saline and dried over magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (eluting solvent: n-hexane-ethyl acetate 1: 1) to obtain 2.62 g of the desired product as a colorless oil.

¹ H-NMR (δ, CDCl ₃ ): 1.99 (s, 3H), 2.05
(s, 3H), 2.06 (s, 3H), 2.15 (s, 3H), 3.40 (t, 2H, J = 5.0H
z), 3.64-3.69 (m, 8H), 3.73-3.78 (m, 1H), 3.90-3.93
(m, 1H), 3.94-3.98 (m, 1H), 4.12 (dd, 1H, J = 6.8Hz, 11.2H
z), 4.18 (dd, 1H, J = 6.3Hz, 11.2Hz), 4.57 (d, 1H, J = 8.1H
z), 5.02 (dd, 1H, J = 3.4Hz, 10.5Hz), 5.21 (dd, 1H, J = 8.1H
z, 10.5Hz), 5.39 (dd, 1H, J = 1.0Hz, 3.4Hz).

[Α] _D ²⁰ = -8.1 ° (c = 1.03, CHCl ₃ ).

2) Synthesis of compound 226 To 0.928 g of compound 225, 70 ml of ethyl acetate was added and dissolved. 0.350 g of p-toluenesulfonic acid monohydrate and 0.506 g of Lindlar's catalyst were added thereto and catalytically reduced at 50 psi for 4 hours. Add 0.509 g of Lindlar's catalyst to 50
Catalytic reduction was performed at psi for 6 hours. The catalyst was filtered off, and 1.001 g of the desired product was obtained as a light brown oily substance. It was used in the following reaction without further purification.

3) Synthesis of Compound 227 20 ml of methylene chloride was added to 1.001 g of Compound 226, 214 μl of triethylamine was added thereto to make a uniform solution, and the mixture was stirred under ice cooling. 0.815 g of (N-palmitoyloxy) succinimide was dissolved in 6 ml of methylene chloride and added, and the mixture was stirred for 3 days while warming to room temperature. The solvent was distilled off under reduced pressure, and the residue was purified by silica gel chromatography (elution solvent: n-hexane-ethyl acetate 1:
1), 0.38 g of the desired product was obtained.

¹ H-NMR (δ, CDCl ₃ ): 0.87 (t, 3H, J = 7.0H
z), 1.16-1.32 (m, 24H), 1.59-1.65 (m, 2H), 1.99 (s, 3
H), 2.05 (s, 3H) 2.06 (s, 3H), 2.15 (s, 3H), 2.18 (t, 2H,
J = 7.6Hz), 3.44-3.48 (m, 2H), 3.55 (t, 2H, J = 5.3Hz), 3.
59-3.67 (m, 6H), 3.72-3.76 (m, 1H), 3.90-3.93 (m, 1H)
， 3.97-4.00 (m, 1H) ， 4.13 (dd, 1H, J = 6.8Hz, 11.2Hz) ，
4.18 (dd, 1H, J = 6.3Hz, 11.2Hz), 4.55 (d, 1H, J = 7.8Hz), 5.
02 (dd, 1H, J = 3.3Hz, 10.3Hz), 5.21 (dd, 1H, J = 7.8Hz, 10.3H
z), 5.39 (dd, 1H, J = 0.5Hz, 3.3Hz), 6.04 (bs, 1H).

[Α] _D ²³ = -7.1 ° (c = 1.02, CHCl ₃ ).

4) Synthesis of Compound 228 To 1.42 g of Compound 227, 20 ml of methanol was added and stirred under ice cooling. Six drops of 28% sodium methoxide methanol solution was added thereto to adjust the pH to 12, and the mixture was stirred at room temperature for 12.5 hours. "Dowex 50X-8" ion exchange resin (H type) was added thereto for neutralization, and the resin was filtered off. The solvent was evaporated under reduced pressure, and the residue was purified by "Sephadex LH-20" (eluting solvent: chloroform-methanol 1: 1) to obtain 1.11 g of the target compound.

¹ H-NMR (δ, pyridine-d ₅ -D ₂ O): 0.87 (t,
3H, J = 7.0Hz), 1.23-1.38 (m, 24H), 1.80 (quintet, 2H, J =
7.6Hz), 2.40 (t, 1H, J = 7.6Hz), 3.55-3.57 (m, 2H), 3.58
-3.60 (m, 6H), 3.63-3.71 (m, 6H), 3.89 (dt, 1H, J = 5.3H
z, 10.6Hz), 4.02-4.05 (m, 1H), 4.13 (dd, 1H, J = 3.4Hz, 9.5
Hz), 4.25 (dt, 1H, J = 5.3Hz, 10.6Hz), 4.41-4.44 (m, 3H)
, 4.54 (bd, 1H), 4.78 (d, 1H, J = 7.6Hz), 8.54 (bt, 1H)
.

[Α] _D ²⁵ = -1.7 ° (c = 1.00, CHCl ₃ -MeOH 1: 1).

FAB-MS: [M + H] ⁺ ; m / z = 550.

(B ′) Synthesis of Compound 851 (FIG. 6) A solution of the para-toluenesulfonate of the amino compound 818 (167 mg) in methylene chloride (10 ml) was added to a solution of N-palmitoyloxysuccinimide (115 mg) in toluene (5 ml). Triethylamine (90 μl) was added, and the mixture was stirred overnight. The reaction solution was washed with water and a half-saturated saline solution, and dried with magnesium sulfate. The solvent was distilled off, and the residue was purified by silica gel column chromatography to obtain a palmitoyl compound (181 mg).

[Α] _D ²³ −20.6 ° (c 1.2, chloroform).

IR (KBr): 3700, 3600, 2980, 2435, 152
0, 1480, 1420 cm ^-1 .

¹ H-NMR (CD ₃ OD): 0.90 (3H, t, J = 7Hz), 1.9
3 (3H, s), 1.95 (3H, s), 2.02 (3H, s), 2.14 (3H, s), 4.46
(1H, d, J = 4.6Hz), 5.06 (1H, dd, J = 3.5,11.5Hz), 5.33 (1H,
brs).

R _f = 0.4 (chloroform: methanol = 93: 7).

Sodium methoxide (40% of a 28% methanol solution) was added to a solution of the palmitoyl compound (141 mg) obtained above in methanol (15 ml), and the mixture was stirred at room temperature for 5.5 hours. The cation exchange resin "Amberlyst 15E" (made by Rohm and Haas) was added until the liquid became neutral, the resin was filtered off, and the filtrate was concentrated to obtain the target compound 851 (10
4 mg, 90%) was obtained.

[Α] _D ²⁶ + 43.9 ° (c 1.0, methanol).

IR (KBr): 3340, 3330, 1640, 1560, 1470 ^-1 cm.

¹ H-NMR (CD ₃ OD): 0.90 (3H, t, J = 7Hz), 1.9
8 (3H, s), 2.19 (2Ht, J = 7Hz,), 3.83 (1H, d, J = 3Hz), 3.90
(1H, t, J = 8.5Hz), 4.44 (1H, d, J = 8.5Hz).

R _f = 0.26 (chloroform: methanol = 9: 1).

(C ′) Synthesis of Compound 333 (FIG. 7) 1) Synthesis of Compound 332 Amine compound 329 (210 mg) and palmitic acid N-hydroxysuccinimide ester (113 mg) were dissolved in methylene chloride (20 ml) to give triethylamine. (65 mg) was added, and the mixture was stirred at room temperature for 4 hours. The reaction solution was diluted with methylene chloride, washed successively with water, 10% citric acid water and water, dried and the solvent was evaporated under reduced pressure. The residue was purified by column chromatography (chloroform-methanol 100: 1) using silica gel (50 g) to obtain the target compound 332 (194 mg).

[Α] _D + 23.7 ° (c 1.07, CHCl ₃ ).

¹ H-NMR (CDCl ₃ ) δ: 0.88 (3H, t, J = 6.8H
z), 1.23-1.33 (24H, m), 1.63 (2H, m), 2.00 (3H, s), 2.04
(3H, s), 2.11 (3H, s), 2.16 (3H, s), 2.17 (2H, t, J = 7.8H
z), 3.45 (2H, q), 3.56 (2H, t), 3.60-3.73 (7H, m), 3.80
-3.85 (1H, m), 4.07 (1H, ddd, J = 2.4Hz, 4.9Hz, 10.0Hz),
4.11 (1H, dd, J = 2.4Hz, 12.2Hz), 4.28 (1H, dd, J = 4.9Hz, 12.
2Hz), 4.89 (1H, d, J = 1.5Hz), 5.27 (1H, dd, J = 1.5Hz, 3.4H
z), 5.30 (1H, t, J = 10.0Hz), 5.36 (1H, dd, J = 3.4Hz, 10.0H)
z), 6.06 (1H, m).

2) Synthesis of Compound 333 Compound 332 (185 mg) was dissolved in methanol (5 ml),
28% NaOMe in MeOH (20 μl) was added, and the mixture was stirred at room temperature for 4 hours. "Amberlite IRC-
50 "was added, the insoluble material was filtered off, and the filtrate was concentrated to dryness under reduced pressure. Chloroform was added to the residue, the insoluble material was filtered off, the filtrate was concentrated to dryness under reduced pressure, and the residue was washed with ether to obtain the target compound 333 (128 mg) as a colorless powder.

[Α] _D + 28.2 ° (c 1.02, CHCl ₃ ).

¹ H-NMR (CDCl ₃ ) δ: 0.88 (3H, t, J = 6.6H
z), 1.20-1.34 (24H, m), 1.62 (2H, m), 2.18 (2H, t, J = 7.6H
z), 3.42-3.93 (18H, m), 4.88 (1H, s), 6.31 (1H, m).

(D ′) Synthesis of Compound 529 (FIG. 8) 1) Synthesis of Compound 524 L-fucose tetraacetate (Compound 523) (10.16)
g) and 2- [2- (2-chloroethoxy) ethoxy] ethanol (10.31 g) were dissolved in methylene chloride (300 ml), BF ₃ .Et ₂ O (17.36 g) was added, and the mixture was stirred at room temperature for 25 hours. . Dilute the reaction mixture with methylene chloride, water, 5%
It was washed successively with NaHCO ₃ water and water, dried and the solvent was evaporated under reduced pressure. The residue was dissolved in pyridine (26 ml), acetic anhydride (20 ml) was added, and the mixture was stirred at room temperature for 15 hr. The reaction mixture was diluted with ethyl acetate, water, 5% NaHCO ₃ water, water,
The mixture was washed successively with 10% citric acid water and water, dried and the solvent was evaporated under reduced pressure. The residue was purified by column chromatography (chloroform) using silica gel (250 g) to obtain α-glycoside compound 524 (4.40 g, 33%) and β-glycoside compound 525 (1.79 g) as colorless oils.

Α-Glycoside compound 524: [α] _D -112.3 ° (c 0.96, CHCl ₃ ).

¹ H-NMR (CDCl ₃ ) δ: 1.14 (3H, d, J = 6.6H
z), 1.99 (3H, s), 2.08 (3H, s), 2.17 (3H, s), 3.62-3.84
(12H, m), 4.23 (1H, q, J = 6.6Hz), 5.11 (1H, d, J = 3.7Hz),
5.12 (1H, dd, J = 3.7Hz, 10.0Hz), 5.30 (1H, d, J = 3.4Hz), 5.
37 (1H, dd, J = 3.4Hz, 10.0Hz).

2) Synthesis of Compound 526 Chloro compound 524 (3.56 g) was dissolved in DMF (50 ml), sodium azide (1.05 g) was added, and the mixture was stirred at 70 ° C. for 2 days. The reaction solution was diluted with ethyl acetate, washed with water, dried and the solvent was evaporated under reduced pressure. The residue is silica gel (100 g)
Was purified by column chromatography (chloroform), and the azide compound 526 (2.98 g) was obtained as a colorless oil.

[Α] _D -113.7 ° (c 0.96, CHCl ₃ ).

¹ H-NMR (CDCl ₃ ) δ: 1.14 (3H, d, J = 6.6H
z), 1.99 (3H, s), 2.07 (3H, s), 2.16 (3H, s), 3.40 (2H, t,
J = 5.0Hz), 3.62-3.70 (9H, m), 3.78-3.84 (1H, m), 4.23
(1H, dq, J = 1.2Hz, 6.6Hz), 5.10 (1H, d, J = 3.7Hz), 5.12 (1
H, dd, J = 3.7Hz, 10.3Hz), 5.30 (1H, dd, J = 1.2Hz, 3.4Hz),
5.37 (1H, dd, J = 3.4Hz, 10.3Hz).

3) Synthesis of compound 527 Azide compound 526 (2.21 g) and p-toluenesulfonic acid (0.94 g) were dissolved in ethanol (100 ml), Lindlar catalyst (4.40 g) was added, and the mixture was heated at room temperature of 50 psi for 7 hours. Catalytic reduction was performed. After the catalyst was filtered off, the filtrate was concentrated under reduced pressure to give amine compound 527 (2.84 g) as a colorless oily substance.

4) Synthesis of compound 528 Amine compound 527 (240 mg) and palmitic acid N-hydroxysuccinimide ester (143 mg) were dissolved in methylene chloride (20 ml), triethylamine (82 mg) was added,
Stir at room temperature for 3 hours. The reaction solution was diluted with methylene chloride, washed successively with water, 10% citric acid water and water, dried and the solvent was evaporated under reduced pressure. The residue was purified by column chromatography (chloroform-methanol 100: 1) using silica gel (50 g) to obtain the target compound (241 mg).

[Α] _D −65.0 ° (c 1.01, CHCl ₃ ).

1H-NMR (CDCl ₃ ) δ: 0.88 (3H, t, J = 6.8H
z), 1.14 (3H, d, J = 6.6Hz), 1.20-1.34 (24H, m), 1.63 (2H,
m), 1.99 (3H, s), 2.07 (3H, s), 2.17 (3H, s), 2.18 (2H,
m), 3.46 (2H, m), 3.56 (2H, m), 3.58-3.68 (7H, m), 3.80
(1H, m), 4.23 (1H, dq, J = 1.2Hz, 6.6Hz), 5.12 (1H, dd, J =
3.7Hz, 10.3Hz), 5.13 (1H, d, J = 3.7Hz), 5.29 (1H, dd, J = 1.
2Hz, 3.4Hz), 5.37 (1H, dd, J = 3.4Hz, 0.3Hz), 6.10 (1H,
m).

5) Synthesis of Compound 529 Compound 528 (208 mg) was dissolved in methanol (5 ml),
28% NaOMe in MeOH (20 μl) was added, and the mixture was stirred at room temperature for 2.5 hours. "Amberlite IRC"
-50 "was added, the insoluble material was filtered off, and the filtrate was concentrated to dryness under reduced pressure. Chloroform was added to the residue, the insoluble material was filtered off, the filtrate was concentrated to dryness under reduced pressure, and the residue was washed with ether to give the object compound 529 (151 mg) as a colorless powder.

[Α] _D -55.2 ° (c 0.56, CHCl ₃ ).

¹ H-NMR (CDCl ₃ ) δ: 0.88 (3H, t, J = 6.8H
z), 1.20-1.34 (24H, m), 1.30 (3H, d, J = 6.6Hz), 1.62 (2H,
m), 2.18 (2H, m), 3.40-3.72 (11H, m), 3.74-3.84 (3H, m)
, 3.90-3.94 (1H, m), 4.04 (1H, q, J = 3.3Hz), 4.92 (1H,
d, J = 3.4Hz), 6.43 (1H, m).

Example 2 (Synthesis Example (2)) A polyethylene glycol having a degree of polymerization of 3 to 6 and at least two alkyl groups having 5 to 20 carbon atoms are contained in the molecule.
Compounds of the invention (invention) 243, 251, 830, 833, 856, 7
18, 1105, 1205, 4-3, 8-2, 12-8, 15-2, 2
8-2 and 18-2 were respectively synthesized as follows.

The reactions in these synthetic reactions are shown in FIGS.
Shown in.

(E) Synthesis of Compound 243 (FIG. 9) 1) Synthesis of Compound 240 J. Org, Chem. , 56 , 4326 (1991), synthesized by the method of 1,2- (2- [2- (2-azidoekito) ethoxy] ethoxy} ethanol (1,980 g) and β-D-galacto-spentaacetate (3.525 g) in methylene chloride (50 ml).
And was stirred under ice-cooling. 4.44 ml of boron trifluoride diethyl ether complex was added dropwise thereto. After stirring at room temperature for 17.5 hours, the mixture was poured into ice water and the organic layer was separated. The extract was washed 5 times with saturated brine, dried over magnesium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (elution solvent: n-hexane-ethyl acetate 1: 1:
5), 1.238 g of the desired product was obtained as a colorless oil.

¹ H-MNR (δ, CDCl ₃ ): 1.99 (s, 3H), 2.05
(s, 3H), 2.06 (s, 3H), 2.15 (s, 3H), 3.40 (t, 2H, J = 5.0H
z), 3.62-3.69 (m, 12H), 3.75 (ddd, 1H, J = 3.7Hz, 7.4Hz, 1
1.1Hz), 3.90-3.93 (m, 1H), 3.94-9.98 (dt, 1H, J = 4.3Hz,
11.1Hz), 4.13 (dd, 1H, J = 6.8Hz, 11.2Hz), 4.18 (dd, 1H, J
= 6.6Hz, 11.2Hz), 4.57 (d, 1H, J = 8.1Hz), 5.02 (dd, 1H, J =
3.4Hz, 10.5Hz), 5.21 (dd, 1H, J = 8.1Hz, 10.5Hz), 5.39 (d
d, 1H, J = 1.0Hz, 3.4Hz).

[Α] _D ²³ = -5.4 ° (c = 1.02, CHCl ₃ ).

2) Synthesis of Compound 241 120 ml of ethyl acetate was added to 1.129 g of compounds 240 to dissolve them. 0.391 g of p-toluenesulfonic acid monohydrate
And 0.570 g of Lindlar's catalyst were added and catalytic reduction was carried out at 50 psi for 5.5 hours. Further, 0.564 g of Lindlar catalyst was added, and the catalyst was reduced at 50 psi for 5 hours. The catalyst was filtered off to obtain 1.172 g of the desired product as a light brown oily substance. It was used for the following reaction without further purification.

3) Synthesis of Compound 1103 20.5 ml of thionyl chloride was added to 0.525 g of 2- (n-hexadecyl) octadecanoic acid, and the mixture was heated under reflux for 6 hours. Thionyl chloride was distilled off under reduced pressure. Benzene was added to the residue to dissolve it, and the residue was filtered off under reduced pressure (3 times). It was used in the following reaction without further purification.

4) Synthesis of compound 242 To 0.618 g of compound 241, 10 ml of methylene chloride was added and dissolved, and the mixture was stirred under ice cooling. 268 μl of triethylamine
Then, the whole amount of the compound 1103 obtained in the above reaction was dissolved in 4 ml of methylene chloride and added, and the mixture was stirred for 14 hours while warming to room temperature. It was diluted with methylene chloride, washed with water and saturated brine, dried over magnesium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (eluting solvent: n-hexane-ethyl acetate 2: 3),
0.550 g of the desired product was obtained.

¹ H-NMR (δ, CDCl ₃ ): 0.88 (t, 6H, J = 7.0H
z), 1.19-1.33 (m, 56H), 1.35-1.42 (m, 2H), 1.53-1.62
(m, 2H), 1.97-2.03 (m, 1H), 1.99 (s, 3H), 2.05 (s, 3
H), 2.06 (s, 3H), 2.15 (s, 3H), 3.45-3.48 (m, 1H), 3.48
-3.54 (t, 2H, J = 5.0Hz), 3.61-3.68 (m, 10H), 3.75 (ddd, 1
H, J = 3.7Hz, 7.3Hz, 11.0Hz), 3.90-3.94 (m, 1H), 3.96 (d
t, 1H, J = 4.4Hz), 4.13 (dd, 1H, J = 7.1Hz, 11.2Hz), 4.17 (d
d, 1H, J = 6.6Hz, 11.2Hz), 4.56 (d, 1H, J = 8.1Hz), 5.02 (dd,
1H, J = 3.5Hz, 10.5Hz), 5.21 (dd, 1H, J = 8.1Hz, 10.5Hz), 5.
39 (dd, 1H, J = 1.0Hz, 3.5Hz), 5.96 (t, 1H, J = 5.6Hz).

[Α] _D ²³ = -2.5 ° (c = 1.00, CHCl ₃ -MeOH 1: 1).

5) Synthesis of Compound 243 To 0.518 g of Compound 242, 5 ml of methanol and 10 parts of benzene were added.
ml was added and dissolved. To this, 6 drops of 28% sodium methoxide methanol solution was added to adjust the pH to 10, and the mixture was stirred at room temperature for 19 hours. "Dowex 50X-8" ion exchange resin (H type) was added thereto for neutralization, and the resin was filtered off. The solvent was distilled off under reduced pressure, and the residue was purified by "Sephadex LH-20" (elution solvent: chloroform-methanol 1:
1), 0.403 g of the target compound was obtained.

¹ H-NMR (δ, pyridine-d ₅ -D ₂ O): 0.88 (t,
6H, J = 7.0Hz), 1.02-1.39 (m, 52H), 1.44-1.62 (m, 6H),
1.92-2.00 (m, 2H), 2.51-2.57 (m, 1H), 3.63-3.77 (m, 14
H), 3.93 (dt, 1H, J = 5.3Hz, 10.7Hz), 4.02-4.05 (m, 1H),
4.14 (dd, 1H, J = 3.4Hz, 9.5Hz), 4.26 (dt, 1H, J = 4.9Hz, 10.7
Hz), 4.40-4.44 (m, 3H), 4.54 (bd, 1H), 4.78 (d, 1H, J
_{1, 2} = 7.6Hz), 8.776 (bt, 1H).

[Α] _D ²⁸ = -3.6 ° (c = 1.00, CHCl ₃ -MeOH 1: 1).

FAB-MS: [M + H] ⁺ ; m / z = 846.

(F) Synthesis of Compound 251 (FIG. 10) 1) Synthesis of Compound HHH Sodium hydride (1.55 g, 60% dispersion) was washed with n-hexane and suspended in 40 ml of N, N-dimethylformamide. The mixture was stirred under ice cooling. Malonic acid dibenzyl ester (4.47 ml) was dissolved in N, N-dimethylformamide (10 ml) and added dropwise, and the mixture was stirred at room temperature for 30 minutes. The mixture was ice-cooled again, 7.29 ml of n-octyl bromide was added, and the mixture was stirred at 60 ° C. for 18 hours.
The solvent was filtered off under reduced pressure. Ethyl acetate and water were added to the residue,
The organic layer was separated. The extract was washed with saturated brine, dried over magnesium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (elution solvent: n-
Hexane-ethyl acetate 20: 1) and the target product was obtained as a colorless oil (7.227 g).

¹ H-NMR (δ, CDCl ₃ ): 0.87 (t, 6H, J = 7.1H
z), 1.02-1.09 (m, 4H), 1.15-1.30 (m, 20H), 1.86-1.89
(m, 4H), 5.10 (s, 4H), 5.10 (s, 4H), 7.25-7.32 (m, 10
H).

2) Synthesis of Compound JJJ To 70 mg of ethyl acetate was added to 6.967 g of compound HHH to dissolve it, 0.105 g (dry matter) of 10% Pd-C was added, and the mixture was subjected to catalytic reduction under normal pressure for 11.5 hours. The catalyst was filtered off, and the solvent was distilled off under reduced pressure. Compound III was obtained as a colorless powder. This was heated at 140 ° C. for 2.5 hours under an argon atmosphere. Allow to cool and compound JJJ
Was obtained as colorless crystals (3.824 g).

Compound III : ¹ H-NMR (δ, CDCl ₃ ): 0.
87 (t, 6H, J = 7.1Hz), 1.20-1.32 (m, 24H), 1.94-1.97 (m, 4
H).

Compound JJ : ¹ H-NMR (δ, CDCl ₃ ): 0.
88 (t, 6H, J = 7.0Hz), 1.21-1.34 (m, 24H), 1.43-1.51 (m, 2
H), 1.58-1.64 (m, 2H), 2.33-2.38 (m, 1H).

3) Synthesis of Compound KKK To 0.336 g of Compound JJJ , 1.5 ml of thionyl chloride was added to give 80%.
Heated at ℃ for 2.5 hours. Thionyl chloride was distilled off under reduced pressure. Benzene was added to the residue to dissolve it, and the residue was evaporated under reduced pressure (three times). It was used in the following reaction without further purification.

4) Synthesis of Compound 250 To 0.442 g of Compound 226, 5 ml of methylene chloride was added and dissolved, and the mixture was stirred under ice cooling. 259 μl of triethylamine
Was added, and the entire amount of the compound KKK obtained in the above reaction was added by dissolving 5 ml of methylene chloride, and the mixture was stirred for 12 hours while warming to room temperature. It was diluted with chloroform, washed with water and saturated brine, dried over magnesium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel chromatography (eluting solvent: n-hexane-ethyl acetate 1: 2) to obtain 0.357 g of the desired product as a colorless amorphous substance.

¹ H-NMR (δ, CDCl ₃ ): 0.88 (t, 6H, J = 7.0H
z), 1.17-1.32 (m, 24H), 1.36-1.43 (m, 2H), 1.53-1.62
(m, 2H), 1.97-2.03 (m, 1H), 1.99 (s, 3H), 2.05 (s, 3
H), 2.06 (s, 3H), 2.15 (s, 3H), 3.46-3.49 (m, 2H), 3.54
(t, 2H, J = 5.0Hz), 3.58-3.69 (m, 6H), 3.74 (ddd, 1H, J = 4.
0Hz, 6.7Hz, 10.7Hz), 3.91 (brt, 1H), 3.98 (dt, 1H, J = 4.4
Hz, 10.7Hz), 4.13 (dd, 1H, J = 7.0Hz, 11.3Hz), 4.18 (dd, 1
H, J = 6.6Hz, 11.3Hz), 4.55 (d, 1H, J = 8.0Hz), 5.02 (dd, 1H,
J = 3.4Hz, 10.5Hz), 5.21 (dd, 1H, J = 8.0Hz, 10.5Hz), 5.39
(dd, 1H, J = 1.0Hz), 5.96 (t, 1H, J = 5.5Hz).

[Α] _D ²⁵ = -5.6 ° (c = 0.98, CHCl ₃ ).

5) Synthesis of compound 251 Compound 250, 0.325 g, methanol 10 ml and benzene 5
ml was added and dissolved. Four drops of 28% sodium methoxide methanol solution was added to adjust the pH to 11, and the mixture was stirred at room temperature for 14 hours. "Dowex 50X-8" ion exchange resin (H type) was added thereto for neutralization, and the resin was filtered off. The solvent was distilled off under reduced pressure, and the residue was purified by "Sephadex LH-20" (elution solvent: chloroform-methanol 1:
1), 0.235 g of the target compound was obtained as a colorless amorphous substance.

¹ H-NMR (δ, pyridine-d ₅ -D ₂ O): 0.85 (t,
6H, J = 7.0Hz), 1.16-1.38 (m, 20H), 1.40-1.58 (m, 6H),
1.89-1.98 (m, 2H), 2.49-2.55 (m, 1H), 3.60-3.65 (m, 4
H), 3.69-3.77 (m, 6H), 3.93 (dt, 1H, J = 5.3Hz, 10.7H
z), 405 (brt, 1H), 4.15 (dd, 1H, J = 3.4Hz), 4.27 (dt, 1
H, J = 5.0Hz, 10.7Hz), 4.42 (d, 2H, J = 6.1Hz), 4.44 (dd, 1H,
J = 7.6Hz, 9.5Hz), 4.55 (br d, 1H), 4.79 (d, 1H, J = 7.6H
z), 8.75 (brt, 1H).

[Α] _D ²⁵ = -2.0 ° (c = 0.98, CHCl ₃ -MeOH 1: 1).

FAB-MS: [M + H] ⁺ ; m / z = 578.

(G) Synthesis of Compound 830 (FIG. 11 ) To a solution of amide compound 829 (87 mg) in methanol (4 ml) was added sodium methoxide (8 μl, 28% methanol solution), and the mixture was stirred at room temperature for 1.5 hours for reaction. Let

The reaction solution was centrifuged, methanol was added to the obtained insoluble matter, and the mixture was centrifuged again to obtain GalNA.
49 mg (yield 67%) of the c derivative compound 830 was obtained.

[0150] R _F 0.37 (chloroform - methanol - water 10: 6: 1).

¹ H-NMR (CD ₃ OD + CDCl ₃ ) δ (ppm) in 500
MHz: 0.89 (s, 6H), 2.01 (brs, 9H), 4.45-4.50 (m, 3H).

IR (KBr): 3400, 3300, 1650, 1560 cm ^-1 .

(H) Synthesis of Compound 833 (FIG. 12) Sodium methoxide (6 μl, 28% methanol solution) was added to a solution of amide compound 832 (69 mg) in methanol (3 ml), and the mixture was stirred at room temperature for 3 hours for reaction. Let

The reaction solution was centrifuged, methanol was added to the obtained insoluble matter, and the mixture was centrifuged again to obtain GalNA.
42 mg (yield 73%) of the c derivative compound 833 was obtained.

[0155] R _F 0.35 (chloroform - methanol - water 10: 6: 1).

[Α] _D −2.6 ° (c 0.58, chloroform-methanol-water 10: 10: 3).

¹ H-NMR (CD ₃ OD + CDCl ₃ ) δ (ppm) in 500
MHz: 0.89 (t, 3H), 2.02 (s, 9H), 4.4-4.5 (m, 3H).

IR (KBr): 3400, 3300, 1650, 1550 cm ^-1 .

(I) Synthesis of Compound 852 (FIG. 13) 1) Synthesis of Compound 1103 To 0.291 g of 2- (n-hexadecyl) octadecanoic acid was added 2 ml of thionyl chloride, and the mixture was heated under reflux for 6 hours. Thionyl chloride was distilled off under reduced pressure. Benzene was added to the residue to dissolve it, and the residue was filtered off under reduced pressure. (Twice). It was used in the following reaction without further purification.

2) Synthesis of Compound 855 To 0.278 g of Compound 818, 5 ml of methylene chloride was added and dissolved, and the mixture was stirred under ice cooling. 139 μl of triethylamine
Then, the whole amount of the compound 1103 obtained in the above reaction was dissolved in 5 ml of methylene chloride and added, and the mixture was stirred for 3 days while warming to room temperature. It was diluted with methylene chloride, washed with water and saturated brine, dried over magnesium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (elution solvent: methylene chloride-methanol 70:
1), 0.318 g of the desired product was obtained as colorless amorphous.

The ¹ H-NMR shows a mixture of two conformers of about 1: 1.
One hydrogen of the conformer is counted and shown as 1H.

¹ H-NMR (δ, CD ₃ OD): 0.90 (t, 12H, J = 7.0H
z), 1.20-1.42 (m, 116H), 1.51-1.59 (m, 4H), 1.93 (s,
3H), 1.94 (2,3H), 1.95 (s, 6H), 2.03 (s, 3H), 2.14 (s, 3
H), 2.15-2.21 (m, 2H), 3.34-3.41 (m, 4H), 3.36 (t, 2H,
J = 5.6Hz), 3.57-5.76 (m, 10H), 3.90-3.96 (m, 2H), 4.02
(brt, 2H), 4.07-4.19 (m, 6H), 4.64 (d, 1H, J = 8.5Hz), 4.
65 (d, 1H, J = 8.5Hz), 5.05 (dd, 1H, J = 9.5Hz, 3.4Hz), 5.08
(dd, 1H, J = 9.4Hz, 3.3Hz), 5.33 (brd, 1H), 5.34 (brd, 1H)
.

[Α] _D ²⁶ = -12.2 ° (c = 1.00, CHCl ₃ -MeOH 1: 1).

3) Synthesis of Compound 856 (GalNAc-t-pas) Compound 855, 0.300 g, was mixed with 4 ml of benzene and 2 parts of methanol.
ml was added and dissolved, and the mixture was stirred under ice cooling. To this, 3 drops of 28% sodium methoxide methanol solution was added to adjust pH to 12, and the mixture was stirred at room temperature for 12 hours. Here, "Dowex 50X
-8 "ion exchange resin (H type) was added for neutralization, and the resin was filtered off. The solvent was distilled off under reduced pressure, the residue was purified by "Sephadex LH-20" (eluting solvent: chloroform-methanol 1: 1), and the target compound was colorless powder 0.147 g.
Obtained.

¹ H-NMR (δ, pyridine-d ₅ -D ₂ O): 0.88 (t,
3H, J = 7.0Hz), 1.19-1.41 (m, 52H), 1.43-1.62 (m, 6H),
1.92-2.00 (m, 2H), 2.13 (s, 3H), 2.52-2.59 (m, 1H), 3.
63-3.80 (m, 10H), 3.91 (dt, 1H, J = 5.4Hz), 3.99-4.01 (br
t, 1H), 4.19 (dt, 1H, H = 4.6Hz, 11.0Hz), 4.33 (dd, 1H, J =
3.2Hz, 10.6Hz), 4.37-4.42 (m, 3H), 4.49 (dm1H, J = 3.2H
z), 4.82 (dd, 1H, J = 8.4Hz, 10.6Hz), 5.05 (d, 1H, J = 8.4H
z), 8.84 (br t, 1H).

[Α] _D ²⁵ = -5.9 ° (c = 0.99, CHCl ₃ -MeOH 1: 1).

FAB-MS: [M + H] ⁺ ; m / z = 843.

(J) Synthesis of Compound 718 (FIG. 14) 1) Synthesis of Compound 715 β-D-lacto-soctaacetate (Compound 701) 6.
99 g and 2- [2- (2-azidoethoxy) ethoxy]
2.35 g of ethanol was dissolved in 40 ml of methylene chloride, and the mixture was stirred under ice cooling. Here, boron trifluoride diethyl ether complex 5.
1 ml was dissolved in 15 ml of methylene chloride and added dropwise over 15 minutes.
After stirring at room temperature for 15 hours, the mixture was poured into ice water and the organic layer was separated. After washing three times with water (the aqueous layer became neutral), the mixture was washed with saturated saline and dried over magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (elution solvent: n-hexane-ethyl acetate 1: 1:
2), 2.87 g of the desired product was obtained as a colorless oil.

¹ H-NMR (δ, CDCl ₃ ): 1.97 (s, 3H), 2.04
(s, 9H), 2.06 (s, 3H), 2.12 (s, 3H), 2.15 (s, 3H), 3.40
(t, 2H, J = 5.0Hz), 3.58-3.75 (m, 10H), 3.79 (t, 1H, J = 9.4H
z), 3.85-3.88 (m, 1H), 3.89-3.93 (m, 1H), 4.06-4.15
(m, 2H), 4.47-4.50 (m, 2H), 4.57 (d, 1H, J = 8.0Hz), 4.9
0 (dd, 1H, J = 8.0Hz, 9.5Hz), 4.95 (dd, 1H, J = 3.5Hz, 10.3H
z), 5.11 (dd, 1H, J = 8.0Hz, 10.3Hz), 5.20 (t, 1H, J = 9.4H
z), 5.35 (bd, 1H, J = 3.5Hz).

[Α] _D ²⁴ = −9.8 ° (c = 1.03, CHCl ₃ ).

2) Synthesis of Compound 716 To 2.69 g of Compound 715, 150 ml of ethyl acetate was added and dissolved. 0.65 g of p-toluenesulfonic acid monohydrate and 1.32 g of Lindlar's catalyst were added thereto and catalytically reduced at 50 psi for 4.5 hours. Furthermore, 1.31 g of Lindlar catalyst is added, 50 ps
It was subjected to catalytic reduction with i for 2.5 hours. The catalyst was filtered off to obtain 3.11 g of the desired product as a light brown oily substance. The following reaction was used without further purification.

4) Synthesis of Compound 717 Compounds 716 and 1.51 g were dissolved by adding 20 ml of methylene chloride and 220 μl of triethylamine, and the whole amount of compound 1103 obtained by the above reaction was dissolved in 5 ml of methylene chloride and added, and further stirred for 19.5 hours. did. It was diluted with methylene chloride, washed with 10% citric acid, water and saturated brine, dried over magnesium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (eluting solvent: n-hexane-ethyl acetate 1: 2) to obtain 1.62 g of the desired product as a colorless oil.

¹ H-NMR (δ, CDCl ₃ ): 0.88 (t, 6H, J = 7.0H
z), 1.19-1.43 (m, 58H), 1.54-1.61 (m, 2H), 1.97 (s, 3
H), 1.97-2.11 (m, 1H), 2.04 (s, 3H), 2.05 (s, 6H), 2.06
(s, 3H), 2.12 (s, 3H), 2.15 (s, 3H), 3.44-3.73 (m, 12H),
3.79 (t, 1H, J = 9.4Hz)), 3.86-3.89 (m, 1H), 3.91-3.95
(m, 1H), 4.06-4.15 (m, 3H), 4.48-4.51 (m, 2H), 4.55
(d, 1H, J = 8.0Hz), 4.90 (dd, 1H, J = 8.0Hz, 9.4Hz), 4.96 (d
d, 1H, J = 3.4Hz, 10.5Hz), 5.11 (dd, 1H, J = 8.1Hz, 10.5Hz),
5.20 (t, 1H, J = 9.4Hz), 5.34 (dd, 1H, J = 0.9Hz, 3.4Hz), 5.
95 (t, 1H, J = 5.5Hz).

[Α] _D ²⁴ = -6.6 ° (c = 1.05, CHCl ₃ ).

5) Synthesis of compound 718 Compound 717 (0.79 g) was added to methanol (5 ml) and benzene (10 ml).
Was added and dissolved, and the mixture was stirred under ice cooling. Then, 5 drops of 28% sodium methoxide methanol solution was added to adjust the pH to 12, and the mixture was stirred at room temperature for 3 hours. Here, "Dowex 50X
-8 "ion exchange resin (H type) was added for neutralization, and the resin was filtered off. The solvent was evaporated under reduced pressure, and the residue was purified by "Sephadex LH-20" (eluting solvent: chloroform-methanol 2: 1) to obtain 0.60 g of the desired compound.

¹ H-NMR (δ, pyridine-d ₅ -D ₂ O): 0.88 (t,
6H, J = 6.8Hz), 1.20-1.61 (m, 58H), 1.90-2.00 (m, 2H),
2.50-2.58 (m, 1H), 3.59-3.82 (m, 11H), 3.84-3.91 (m, 2
H), 3.99 (bt, 1H), 4.10-4.14 (m, 2H), 4.18-4.26 (m, 3
H), 4.34 (dd, 1H, J = 5.0Hz, 11.0Hz), 4.40-4.50 (m, 5H)
, 4.77 (d, 1H, J = 7.5Hz), 5.06 (d, 1H, J = 8.0Hz), 8.82 (b
t, 1H).

[Α] _D ²² = -3.0 ° (c = 1.01, CHCl ₃ -MeOH 2: 1).

FAB-MS: [M + H] ⁺ ; m / z = 964.

(K) Synthesis of Compound 1105 (FIG. 15) 1) Synthesis of Compound 1101 6.16 g of β-D-glucose pentaacetate and 2-
[2- (2-azidoethoxy) ethoxy] ethanol 6.
59 g was dissolved in 50 ml of methylene chloride and stirred under ice cooling. 8.7 ml of boron trifluoride diethyl ether complex was dissolved in 10 ml of methienyl chloride and added dropwise over 5 minutes. After stirring at room temperature for 19 hours, the mixture was poured into ice water and the organic layer was separated. After washing 4 times with water (the water layer became neutral), wash with saturated saline,
It was dried over magnesium sulfate and the solvent was evaporated under reduced pressure.
The residue was purified by silica gel column chromatography (eluting solvent: n-hexane-ethyl acetate 2: 1) to obtain 2.27 g of the desired product as a colorless oil.

¹ H-NMR (δ, CDCl ₃ ): 2.01 (s, 3H), 2.03
(s, 3H), 2.05 (s, 3H), 2.09 (s, 3H), 3.41 (t, 2H, J = 5.0H
z), 3.63-3.78 (m, 10H), 3.93-3.97 (m, 1H), 4.14 (dd, 1
H, J = 2.0Hz, 12.5Hz), 4.26 (dd, 1H, 5.0Hz, 12.5Hz), 4.62
(d, 1H, J = 7.9Hz), 5.00 (dd, 1H, J = 7.9Hz, 9.8Hz), 5.09
(t, J = 9.8Hz), 5.21 (t, 1H, J = 9.8Hz).

[Α] _D ²⁰ = -12.1 ° (c = 1.01, CHCl ₃ ).

2) Synthesis of compound 1102 To 2.11 g of the compounds 1101 and 150 ml of ethyl acetate was added and dissolved. 0.79 g of p-toluenesulfonic acid monohydrate and 1.04 g of Lindlar's catalyst were added thereto and catalytically reduced at 50 psi for 6.5 hours. Furthermore, 1.04 g of Lindlar catalyst is added, and 50 ps
It was catalytically reduced with i for 3 hours. The catalyst was filtered off to obtain 2.49 g of the desired product as a light brown oily substance. Without further purification,
Used for the following reactions.

3) Synthesis of compound 1103 To 0.91 g of 2- (n-hexadecyl) octadecanoic acid was added 5 ml of thionyl chloride, and the mixture was heated under reflux for 6 hours. Thionyl chloride was distilled off under reduced pressure. Benzene was added to the residue to dissolve it, and the residue was evaporated under reduced pressure (twice). It was used in the following reaction without further purification.

4) Synthesis of compound 1104 To 0.90 g of compound 1102, 30 ml of methylene chloride and 190 μl of triethylamine were added and dissolved, and the whole amount of compound 1103 obtained in the above reaction was dissolved in 5 ml of methylene chloride and added, followed by stirring for 2 hours. And further stirred at room temperature for 30 minutes. It was diluted with methylene chloride, washed with 1N hydrochloric acid, water and saturated brine, dried over magnesium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (elution solvent: n-hexane-ethyl acetate 1: 1),
0.67 g of the desired product was obtained as a colorless oil.

¹ H-NMR (δ, CDCl ₃ ): 0.88 (t, 6H, J = 7.0H
z), 1.22-1.43 (m, 58H), 1.56-1.62 (bs, 2H), 1.97-2.02
(m, 1H), 2.01 (s, 3H), 2.03 (s, 3H), 2.05 (s, 3H), 2.09
(s, 3H), 3.44-3.76 (m, 12H), 3.95-3.98 (m, 1H), 4.10-
4.16 (m, 2H), 4.26 (dd, 1H, J = 4.8Hz, 12.3Hz), 4.60 (d, 1
H, J = 8.1Hz), 5.00 (dd, 1H, J = 8.1Hz, 9.5Hz), 5.09 (t, 1H,
J = 9.5Hz), 5.21 (t, 1H, J = 9.5Hz), 5.99 (t, 1H, J = 5.6Hz).

[Α] _D ²¹ = -8.3 ° (c = 1.02, CHCl ₃ ).

5) Synthesis of compound 1105 Compound 0.64 g of compound 1104 was added with 6 ml of methanol and 12 ml of benzene.
Was added and dissolved, and the mixture was stirred under ice cooling. Then, 5 drops of 28% sodium methoxide methanol solution was added to adjust the pH to 12, and the mixture was stirred at room temperature for 5 hours. Here, "Dowex 50X
-8 "ion exchange resin (H type) was added for neutralization, and the resin was filtered off. The solvent was evaporated under reduced pressure, and the residue was purified by "Sephadex LH-20" (eluting solvent: methylene chloride-methanol 1: 1) to obtain 0.46 g of the desired compound.

¹ H-NMR (δ, pyridine-d ₅ -D ₂ O): 0.88 (t,
6H, J = 6.8Hz), 1.22-1.60 (m, 58H), 1.92-1.99 (m, 2H),
2.51-2.58 (m, 1H), 3.59-3.67 (m, 4H), 3.68-3.78 (m, 6
H), 3.90-3.95 (m, 2H), 4.00 (t, 1H, J = 7.8Hz), 4.15-4.
27 (m, 3H), 4.32 (dd, 1H, J = 5.5Hz, 11.7Hz), 4.51 (d, 1H, J
= 11.7Hz), 4.85 (d, 1H, J = 7.8Hz), 8.78 (t, 1H, J = 5.0H)
z).

[Α] _D ²¹ = -7.1 ° (c = 1.02, CHCl ₃ ).

FAB-MS: [M + H] ⁺ ; m / z = 802.

(L) Synthesis of Compound 1205 (FIG. 16) 1) Synthesis of Compound 1202 β-D-ribose tetraacetate (Compound 1201) 4.29
9 g and 2- [2- (2-azidoethoxy) ethoxy]
1.183 g of ethanol was dissolved in 40 ml of methylene chloride, and the mixture was stirred under ice cooling. Here is boron trifluoride diethyl ether complex
3.32 ml was dissolved in 6 ml of methylene chloride and added dropwise. After stirring at room temperature for 1 hour, the mixture was poured into ice water and the organic layer was separated. 6
After being washed with water (the aqueous layer became neutral), it was washed with saturated saline and dried over magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (elution solvent: n-hexane-ethyl acetate 1: 1),
The desired product was obtained as a colorless oily substance (1.967 g).

¹ H-NMR (δ, CDCl ₃ ): 2.06 (s, 3H), 2.09
(s, 3H), 2.11 (s, 3H), 3.40 (t, 2H, J = 5.0Hz), 3.64-3.69
(m, 9H), 3.85 (ddd, 1H, J = 3.1Hz, 5.1Hz, 10.3Hz), 4.14
(dd, 1H, J = 6.0Hz, 11.4Hz), 4.28-4.32 (m, 1H), 4.34 (dd,
1H, J = 3.8Hz, 11.4Hz), 5.06 (s, 1H), 5.28 (d, 1H, J = 4.8H
z), 5.35 (dd, 1H, J = 4.8Hz, 7.0Hz).

[Α] _D ²⁰ = -14.5 ° (c = 1.04, CHCl ₃ ).

2) Synthesis of compound 1203 To 12044 g of compound 1202, 100 ml of ethyl acetate was added and dissolved. 0.809 g of p-toluenesulfonic acid monohydrate
And 0.944 g of Lindlar's catalyst were added, and catalytic reduction was carried out at 50 psi for 4 hours. Furthermore, 0.855 g of Lindlar catalyst is added,
Catalytic reduction was carried out at 50 psi for 3 hours. The catalyst was filtered off, and 2.422 g of the desired product was obtained as a pale brown oily substance. It was used in the following reaction without further purification.

3) Synthesis of compound 1103 To 1.024 g of 2- (n-hexadecyl) octadecanoic acid was added 5 ml of thionyl chloride, and the mixture was heated under reflux for 2.5 hours. Thionyl chloride was distilled off under reduced pressure. Benzene was added to the residue to dissolve it, and the residue was evaporated under reduced pressure (three times). It was used in the following reaction without further purification.

4) Synthesis of compound 1204 Compound 1203 was mixed with 20 ml of methylene chloride and 23 of triethylamine.
6 μl was added and dissolved, and the mixture was stirred under ice cooling. To this, 283 μl of triethylamine was added, and the entire amount of compound 1103 obtained in the above reaction was dissolved in 5 ml of methylene chloride and added, and the mixture was stirred for 17 hours while warming to room temperature. Dilute with methylene chloride,
The extract was washed with water and saturated brine, dried over magnesium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (elution solvent: n-hexane-
Ethyl acetate 3: 2) and 0.910 g of the desired product were obtained.

¹ H-NMR (δ, CDCl ₃ ): 0.88 (t, 6H, J = 7.0H
z), 1.21-1.34 (m, 56H), 1.35-1.44 (m, 2H), 1.56-1.63
(m, 2H), 1.96-2.02 (m, 1H), 2.06 (s, 3H), 2.09 (s, 3
H), 2.11 (s, 3H), 3.45-3.48 (m, 2H), 3.54 (t, 2H, J = 5.0H
z), 3.59-3.66 (m, 7H), 3.84-3.88 (m, 1H), 4.15 (dd, 1)
H, J = 5.3Hz, 10.9Hz), 4.29-4.32 (m, 1H), 4.33 (dd, 1H, J =
4.0Hz, 10.9Hz), 5.06 (s, 1H), 5.28 (d, 1H, J = 5.0Hz),
5.35 (dd, 1H, J = 5.0Hz, 6.7Hz), 6.01 (t, 1H, J = 5.6Hz).

[Α] _D ²⁰ = -6.9 ° (c = 1.03, CHCl ₃ ).

5) Synthesis of compound 1205 To compound 1204, 0.477 g, was added methanol 3 ml and benzene 6
ml was added and dissolved. Add 6 drops of 28% sodium methoxide methanol solution to adjust the pH to 12, and add 1.5 at room temperature.
Stir for hours. "Dowex 50X-8" ion exchange resin (H type) was added thereto for neutralization, and the resin was filtered off. The solvent was distilled off under reduced pressure, and the residue was "Sephadex LH-20".
(Elution solvent: chloroform-methanol 1: 1:
1), 0.389 g of the desired product was obtained.

¹ H-NMR (δ, pyridine-d ₅ -D ₂ O): 0.89 (t,
6H, J = 7.0Hz), 1.20-1.39 (m, 52H), 1.44-1.62 (m, 6H),
1.92-2.01 (m, 2H), 2.55-2.60 (m, 1H), 3.65-3.78 (m, 7
H), 4.09 (ddd, 1H, J = 3.3Hz, 6.0Hz, 10.8Hz), 4.14 (dd, 1
H, J = 5.3Hz, 11.8Hz), 4.27 (dd, 1H, J = 3.3Hz, 11.8Hz) 4.55
(d, 1H, J = 4.9Hz), 4.72-4.75 (m, 1H), 4.84-4.86 (m, 1H)
, 5.48 (s, 1H), 8.84 (bt, 1H).

[Α] _D ²⁰ = -13.3 ° (c = 1.00, CHCl ₃ -MeOH 1: 1).

FAB-MS: [M + H] ⁺ ; m / z = 772.

(M ) Synthesis of Compound 4-3 (FIG. 17) (i) Glycosylation Reaction of Compound 1-1 and Compound 3-1 Compound 1-1 (6.00 g) and alcohol compound (Compound 3-
1) (8.04g) is dissolved in methylene chloride (180ml),
F ₃ · Et ₂ O (9.05 g) was added, and the mixture was stirred at room temperature for 5 days.

The reaction solution was diluted with chloroform and diluted with water, 5%.
It was washed successively with NaHCO ₃ water and water, dried and the solvent was evaporated under reduced pressure. The residue was purified twice by column chromatography (chloroform) using silica gel (200 g) and α-
Glycoside (Compound 3-2) (3.66 g) and β-glycoside (Compound 3-3) (1.60 g) were obtained.

(Α-Glycoside) [α] _D + 92.1 ° (c 0.91, CHCl ₃ ). ¹ H-NMR (CDCl ₃ ) δ: 2.02 (3H, s), 2.0
3 (3H, s), 2.06 (3H, s), 3.63-3.73 (9
H, m), 3.75 (3H, s), 3.77 (2H, m), 3.82
-3.87 (1H, m), 4.45 (1H, d, J = 10.02 H
z), 4.88 (1H, dd, J = 3.67Hz, 10.26H
z), 5.17 (1H, dd, J = 9.77Hz, 10.02H
z), 5.21 (1H, d, J = 3.67Hz), 5.54 (1H,
dd, J = 9.77 Hz, 10.26 Hz). (ii) Synthesis of compound 3-4 Compound 3-2 (3.70 g) was dissolved in DMF (40 ml), sodium azide (0.74 g) was added, and the mixture was stirred at a bath temperature of 60 ° C for 20 hours.

The reaction mixture was diluted with ethyl acetate, washed with water, dried and the solvent was evaporated under reduced pressure. The residue was purified by column chromatography (chloroform) using silica gel (200 g) to obtain the target compound 3-4 (2.10 g) as a colorless oily substance.

¹ H-NMR (CDCl ₃ ) δ: 2.02 (3
H, s), 2.03 (3H, s), 2.06 (3H, s), 3.40
(2H, t, J = 5.13Hz), 3.63-3.73 (9H,
m), 3.75 (3H, s), 3.82-3.87 (1H, m), 4.
45 (1H, d, J = 10.26 Hz), 4.88 (1H, dd,
J = 3.66Hz, 10.26Hz), 5.17 (1H, dd, J =
9.77Hz, 10.26Hz), 5.21 (1H, d, J = 3.66H)
z), 5.54 (1H, dd, J = 9.77Hz, 10.26H
z). (iii) Synthesis of Compound 3-5 Compound 3-4 (1.70 g) and p-toluenesulfonic acid monohydrate (0.66 g) are dissolved in ethanol (100 ml), Lindlar catalyst (3.00 g) is added, and the mixture is allowed to stand at room temperature. Catalytic reduction was carried out at 40 psi for 5 hours.

After filtering the catalyst, the filtrate was concentrated under reduced pressure to obtain the target compound 3-5 (2.05 g) as a colorless foamy substance.

(Iv) Synthesis of Compound 4-2 Carboxylic acid (Compound 4-1) (204 mg), N-hydroxysuccinimide (HOSu) (46 mg) and N, N '.
-Dicyclohexylcarbodiimide (DCC) (83 mg)
A methylene chloride (20 ml) solution of was stirred at room temperature for 20 hours.
The amine compound (compound 3-5) (364 mg) dissolved in methylene chloride (5 ml) was added to the reaction solution, triethylamine (115 mg) was added, and the mixture was stirred at room temperature for 3 hours.

The reaction mixture was diluted with chloroform, water, 5%
The organic layer was washed successively with NaHCO ₃ water, water, citric acid water and water, dried and the solvent was distilled off under reduced pressure. The residue was purified by column chromatography using silica gel (30 g) (hexane-ethyl acetate 1: 1) to obtain the target compound 4-2 (65 mg) as a colorless powder.

[Α] _D + 50.0 ° (c 0.83, CHCl ₃ ). ¹ H-NMR (CDCl ₃ ) δ: 0.88 (6H, t, J =
6.59Hz), 1.20-1.33 (56H, m), 1.34-1.43 (2
H, m), 1.53-1.63 (2H, m), 1.98-2.05 (1
H, m), 2.03 (6H, s), 2.06 (3H, s), 3.45
-3.50 (2H, m ₂ ), 3.25-3.57 (2H, m), 3.58
-3.64 (4H, m), 3.64-3.69 (2H, m), 3.70-
3.76 (1H, m), 3.75 (3H, s), 3.81-3.87 (1
H, m), 4.45 (1H, d, J = 10.26 Hz), 4.88
(1H, dd, J = 3.66Hz, 10.26Hz), 5.17 (1
H, dd, J = 9.28Hz, 10.26Hz), 5.24 (1H,
d, J = 3.66Hz), 5.55 (1H, dd, J = 9.28H
z, 10.26 Hz), 6.04 (1H, m). (v) Synthesis of Compound 4-3 Compound 4-2 (540 mg) was added to methanol (3 ml),
28% NaOMe in MeOH (20 μl) was added, and the mixture was stirred at room temperature for 24 hours. The reaction mixture was concentrated under reduced pressure, methanol (3 ml) and 0.1 N NaOH aqueous solution (2 ml) were added to the residue,
Stir at room temperature for 6 hours.

The reaction solution was ice-cooled and the mother liquor was removed by decantation to obtain the precipitated target compound 4-3 (35 mg) as a colorless powder.

[Α] _D + 23.1 ° (c 0.46, MeOH). ¹ H-NMR (CD ₃ OD) δ: 0.90 (6 H, t, J =
6.84Hz), 1.12-1.34 (56H, m), 1.34-1.45 (2
H, m), 1.50-1.58 (2H, m), 2.19 (1H,
m), 3.34-3.46 (4H, m), 3.52-3.56 (2H,
m), 3.60-3.64 (2H, m), 3.64-3.74 (6H,
m), 3.88-3.92 (1H, m), 3.93 (1H, d, J =
10.26 Hz), 4.86 (1H, d, J = 3.91 Hz). (n) Synthesis of compound 8-2 (Fig. 18) (i) Glycosylation reaction of compound 6-1 and compound 3-1 Compound 6-1 (876 mg) and alcohol (compound 3-1)
(830 mg) was dissolved in methylene chloride (50 ml), and BF ₃ ·
Et ₂ O (932 mg) was added, and the mixture was stirred at room temperature for 21 hours.

The reaction solution was diluted with chloroform, water, 5%
It was washed successively with NaHCO ₃ water and water, dried and the solvent was evaporated under reduced pressure. The residue was subjected to column chromatography using silica gel (150 g) (chloroform-methanol 200: 1).
→ Purify with chloroform-methanol 100: 1), β-
Glycoside (Compound 7-2) (566 mg), followed by β-glycoside (Compound 7-2) and α-glycoside (Compound 7)
A 3: 1 mixture of -1) (208 mg) was obtained.

(Β-Glycoside) [α] _D + 4.6 ° (c 1.75, CHCl ₃ ). ¹ H-NMR (CDCl ₃ ) δ: 1.89 (1 H, dd, J
= 11.70 Hz, 12.95 Hz), 1.89 (3 H, s), 2.01
(3H, s), 2.03 (3H, s), 2.06 (3H, s),
2.15 (3H, s), 2.43 (1H, dd, J = 4.88Hz,
12.95 Hz), 3.51-3.56 (1H, m), 3.62-3.85
(9H, m), 3.80 (3H, s), 3.88-3.97 (2H,
m), 4.12 (1H, dd, J = 8.30Hz, 12.46H
z), 4.13 (1H, ddd, J = 10.26 Hz, 10.51 H
z, 10.99 Hz), 4.56 (1H, dd, J = 2.20 Hz,
10.51 Hz), 4.89 (1H, dd, J = 2.44Hz, 12.4
6 Hz), 5.25 (1H, ddd, J = 4.88Hz, 10.99
Hz, 11.70 Hz), 5.28 (1H, ddd, J = 2.44H
z, 3.42Hz, 8.30Hz), 5.40 (1H, dd, J = 2.
20Hz, 3.42Hz), 5.93 (1H, d, J = 10.26H
z). (ii) Synthesis of compound 7-3 Compound 7-2 (675 mg) was dissolved in DMF (15 ml), sodium azide (137 mg) was added, and the mixture was stirred at a bath temperature of 60 ° C for 40 hours.

The reaction mixture was diluted with ethyl acetate, washed with water,
After drying, the solvent was distilled off under reduced pressure. The residue is treated with silica gel (50
The target compound 7-3 was purified by column chromatography (chloroform-methanol 150: 1) using g).
(510 mg) was obtained as a colorless oily substance.

[Α] _D + 2.6 ° (c 1.02, CHCl ₃ ). ¹ H-NMR (CDCl ₃ ) δ: 1.89 (1 H, dd, J
= 11.70 Hz, 12.94 Hz), 1.89 (3 H, s), 2.01
(3H, s), 2.03 (3H, s), 2.06 (3H, s),
2.15 (3H, s), 2.44 (1H, dd, J = 4.89Hz,
12.94 Hz), 3.43-3.50 (2H, m), 3.50-3.54
(1H, m), 3.61-3.73 (7H, m), 3.75-3.90
(2H, m), 3.80 (3H, s), 4.12 (1H, dd,
J = 8.55Hz, 12.46Hz), 4.14 (1H, ddd, J
= 10.02 Hz, 10.50 Hz, 10.75 Hz), 4.46 (1
H, dd, J = 2.20Hz, 10.50Hz), 4.90 (1H,
dd, J = 2.44 Hz, 12.46 Hz), 5.25 (1H, dd
d, J = 4.89Hz, 10.75Hz, 11.70Hz), 5.27
(1H, ddd, J = 2.44Hz, 3.42Hz, 8.55H
z), 5.40 (1H, dd, J = 2.20Hz, 3.42Hz),
5.94 (1H, d, J = 10.02 Hz). (iii) Synthesis of compound 7-4 Compound 7-3 (510 mg) and p-toluenesulfonic acid monohydrate (150 mg) were dissolved in methanol (50 ml), Lindlar catalyst (1.50 g) was added, and room temperature was 50 psi. The catalytic reduction was performed for 7 hours. After removing the catalyst by filtration, the filtrate was concentrated under reduced pressure to obtain the target compound (590 mg).

(Iv) Synthesis of Compound 8-1 Carboxylic acid (Compound 4-1) (153 mg), N-hydroxysuccinimide (35 mg) and N, N'-dicyclohexylcarbodiimide (62 mg) in methylene chloride (40 ml) A mixed solution of hexane and hexane (20 ml) was stirred at room temperature for 15 hours.
Amine compound 7-4 (200 mg) dissolved in acetonitrile (20 ml) was added to the reaction solution, triethylamine (53 mg) was added, and the mixture was stirred at room temperature for 24 hours. The reaction mixture was concentrated under reduced pressure, diluted with chloroform, water, 5% NaHCO ₃
The extract was washed successively with water, water, citric acid water and water, dried and the solvent was distilled off under reduced pressure. Column chromatography of the residue using silica gel (50 g) (chloroform-methanol)
Purification with 150: 1) gave the target compound 8-1 (162 mg) as a colorless wax.

[Α] _D + 5.9 ° (c 1.03, CHCl ₃ ). ¹ H-NMR (CDCl ₃ ) δ: 0.88 (6H, t, J =
6.84Hz), 1.20-1.33 (56H, m), 1.33-1.44 (2
H, m), 1.53-1.64 (2H, m), 1.86 (3H,
s), 1.90 (1 H, dd, J = 11.48 Hz, 12.94 H
z), 1.99-2.05 (1H, m), 2.01 (3H, s), 2.
03 (3H, s), 2.05 (3H, s), 2.15 (3H,
s), 2.43 (1H, dd, J = 4.89Hz, 12.94H
z), 3.30-3.38 (1H, m), 3.40-3.70 (9H,
m), 3.80 (3H, s), 3.79-3.89 (1H, m), 4.
12 (1H, dd, J = 8.55Hz, 12.46Hz), 4.12
(1H, ddd, J = 10.02 Hz, 10.75 Hz, 10.75
Hz), 4.51 (1H, dd, J = 2.44Hz, 10.75H
z), 4.91 (1H, dd, J = 2.44Hz, 12.46H
z), 5.25 (1H, ddd, J = 2.44Hz, 3.42Hz,
8.55Hz), 5.28 (1H, ddd, J = 4.89Hz, 10.7
5 Hz, 11.48 Hz), 5.41 (1H, dd, J = 2.44H
z, 3.42Hz), 6.28 (1H, m), 6.36 (1H, d,
J = 10.02 Hz). (v) Synthesis of Compound 8-2 Compound 8-1 (155 mg) was dissolved in methanol (4 ml), 28% NaOMe in MeOH (20 μl) was added, and the mixture was stirred at room temperature for 3 hr. The reaction mixture was concentrated under reduced pressure, the residue was dissolved in methanol (12 ml), 0.1 N NaOH aqueous solution (3 ml) was added, and the mixture was stirred at room temperature for 16 hr. After heating the reaction solution, "Amberlite IRC-50" was added, the insoluble matter was filtered off, the filtrate was concentrated to dryness under reduced pressure, and the residue was washed with ether,
The target compound 8-2 (99 mg) was obtained as a colorless powder.

¹ H-NMR (CD ₃ OD) δ: 0.90 (6
H, t, J = 6.59Hz), 1.20-1.44 (58H, m), 1.
50-1.58 (2H, m), 1.63 (1H, dd, J = 11.72)
Hz, 12.95 Hz), 1.99 (3H, s), 2.18 (1H,
m), 2.42 (1H, dd, J = 4.89Hz, 12.95H
z), 3.36-3.40 (2H, m), 3.43-3.47 (1H,
m), 3.51-3.59 (3H, m), 3.60-3.74 (8H,
m), 3.76-3.83 (2H, m), 3.84-3.94 (2H,
m), 3.96-4.02 (1H, m). (o) Synthesis of compound 12-8 (FIG. 19) (i) Synthesis of compound 12-3 1,6-anhydrolactose peracetate (compound
12-1) (50 g, 86.7 mmol), 28% sodium methylate (2 ml) and methanol (600 ml) were stirred at room temperature for 4 hours and treated by a conventional method to prepare 1,6-anhydrolactose (compound 12-2) ( 28.5 g) was obtained.

[Α] _D −46.5 ° (c 1.0, H ₂ O). Compound 12-2 (16.2 g) was dissolved in pyridine (150 ml), trityl chloride (20.9 g) was added, and the mixture was reacted at 50 ° C for 2 hr. Then, at room temperature, a solution of benzoyl chloride in pyridine (5
0.6 g) was added and the mixture was stirred overnight.

The reaction solution was poured into ice water, extracted with chloroform and purified by silica gel column chromatography (ethyl acetate-hexane, 1: 1) to give compound 12-3.
(35 g) was obtained.

Elemental analysis values as C ₆₆ H ₅₄ O ₁₅ were calculated values: C, 72.93; H, 5.01, but actually measured values, C, 72.63;
It was H, 5.30.

[Α] _D + 54.2 ° (c 1.0, CHCl ₃ ). ¹ H-NMR (CDCl ₃ ): 3.16 (1 H, dd, J =
9.5 Hz, 8.8 Hz), 3.35 (1H, dd, J = 9.5H
z, 5.6 Hz), 3.74 (1H, s), 3.76 (1H, d
d, J = 7.6 Hz, 6.0 Hz), 4.01 (1H, d, J =
7.6 Hz), 4.08 (1H, dd, J = 8.8 Hz, 5.6 H
z), 4.53 (1H, d, J = 5.4 Hz), 4.94 (1H,
s), 5.24 (1H, d, J = 8.1 Hz), 5.63 (1H,
dd, J = 10.3Hz, 3.4Hz), 5.64-5.65 (2H,
m), 5.78 (1H, dd, J = 10.3Hz, 8.1Hz),
7.0-8.1 (40H, m). (ii) Synthesis of compound 12-4 Compound 12-3 (34.5 g) was added to chloroform-methanol-
It was dissolved in 400 ml of a mixed solvent of water (100: 100: 1), paratoluenesulfonic acid (1.2 g) was added, and the mixture was reacted at 50 ° C. for 6 hours.

The reaction mixture was neutralized with saturated aqueous sodium hydrogen carbonate, the solvent was evaporated, the mixture was extracted with chloroform and purified by silica gel column chromatography (ethyl acetate-hexane, 1: 1) to give compound 12-4 (18.6 g). ) Got.

The elemental analysis values as C ₄₇ H ₄₀ O _{15 are} as follows: calculated value: C, 66.86; measured value for C, 66.86; C, 66.43;
H, 4.95.

[Α] _D + 104.3 ° (c 1.02, CHCl ₃ ). (iii) Synthesis of Compound 12-5 Compound 12-4 (18.5 g) was dissolved in 90 ml of pyridine and ice-cooled, diphenylphosphorochloridate (8.27 g) was added, and the mixture was returned to room temperature and stirred for 1 hour.

The reaction solution was poured into ice water, extracted with chloroform, and purified by silica gel column chromatography (ethyl acetate-hexane, 1: 1) to give compound 12-
5 (18.6 g) was obtained.

The elemental analysis values as C ₅₉ H ₄₉ O ₁₈ P were the calculated values: C, 65.79; H, 4.59; P, 2.88, whereas the measured values were C, 65.66; H, 4.83; P, 3.24.

[Α] _D + 53.8 ° (c 0.976, CHCl ₃ ). ¹ H-NMR (CDCl ₃ ): 3.78 (2H, m), 4.02
(1H, d, J = 7.6Hz), 4.04 (1H, ddd, J
= 10.5Hz, 7.5Hz, 7.0Hz), 4.22 (1H, dd
d, J = 10.5Hz, 8.0Hz, 7.0Hz), 4.34 (1
H, t, J = 6.0Hz), 4.56 (1H, d, J = 5.4H)
z), 4.96 (1H, s), 5.31 (1H, d, J = 8.1 H
z), 5.58 (1H, dd, J = 10.4Hz, 3.4Hz),
5.67 (2H, m), 5.86 (1H, dd, J = 10.4Hz,
8.1 Hz), 7.0-8.1 (35 H, m). (iv) Synthesis of compound 12-6 A mixture of compound 12-5 (18.2 g), 29 ml of trifluoroacetic acid and 356 ml of acetic anhydride was stirred at room temperature for 20 hours.

The reaction solution was concentrated under reduced pressure, and the residual solid was subjected to silica gel column chromatography (ethyl acetate-hexane,
Compound 12-6 (α / β = 71/2) after purification by 1: 1)
9, 17 g) was obtained.

The elemental analysis values as C ₆₃ H ₅₅ O ₂₁ P were calculated values: C, 64.18; H, 4.70; P, 2.63, but actually measured values:
C, 64.10; H, 4.79; P, 2.88.

[Α] _D + 50.5 ° (c 1.02, CHCl ₃ ). ¹ H-NMR (CDCl ₃ ): (α-anomer); 2.0
3, 2.14 (each s), 3.43 (dt, J = 10.5Hz, 7.0
Hz, 7.0 Hz), 3.63 (dt, J = 10.5Hz, 10.5H
z, 7.0 Hz), 3.90 (t, J = 7.0 Hz), 4.03
(M), 4.06 (t, J = 10.3Hz), 4.22 (dd, J =
11.4Hz, 3.4Hz), 4.27 (dd, J = 11.4Hz, 1.
7 Hz), 4.77 (d, J = 8 Hz), 5.37 (dd, J =
8.5 Hz, 3.5 Hz), 5.67 (dd, J = 10.2 Hz, 7.8
Hz), 5.69 (s), 5.97 (dd, J = 10.3 Hz, 8.5
Hz), 6.47 (d, J = 3.5 Hz), 7.1-8.0
(M). (Β-anomer); 2.02, 2.10 (each s, 2 × OA
c), 3.40 (dt, J = 10.5Hz, 7.0Hz), 3.56
(Dt, J = 10.5Hz, 10.5Hz, 7.0Hz), 3.77
(M), 4.71 (d, J = 7.8 Hz), 5.50 (dd, J =
10.0Hz, 8.3Hz), 5.64 (dd, J = 10.5Hz, 7.
8 Hz), 5.74 (t, J = 9.5 Hz), 5.86 (d, J =
8.3 Hz), 7.1-8.0 (m). (v) Synthesis of compound 12-7 A mixture of compound 12-6 (10 g), triethylene glycol n-octadecyl ether (3.4 g), methylene chloride 150 ml and "Molecular Sieve 4A" (MS4A) was ice-cooled and trimethylsilyl triflate. The rate (2.82 g) was added dropwise, and the mixture was reacted at room temperature for 3 hours.

The reaction solution was poured into 10% aqueous sodium hydrogen carbonate, and the organic layer was purified by silica gel column chromatography (benzene-ethyl acetate-hexane, 1: 1: 1) to give compound 12
-7 (2.7 g) was obtained.

Mp: 39-40 ° C. The elemental analysis value as C ₈₅ H ₁₀₁ O ₂₃ P was calculated value: C,
67.08 ； H, 6.69 Actual value: C, 66.65 ； H, 6.72
Met.

[Α] _D + 18.4 ° (c 0.964, CHCl ₃ ). ¹ H-NMR (CDCl ₃ ): 0.89 (3H, t, J = 7.0
Hz), 1.27 (30H, s), 1.57 (2H, m), 2.00
(3H, s), 3.3-3.7 (14H, m), 3.65 (1H,
ddd, J = 9.4 Hz, 5.0 Hz, 1.7 Hz), 3.98
(1H, t, J = 9.4 Hz), 4.15 (1H, dd, J =
12.0Hz, 5.0Hz), 4.32 (1H, dd, J = 12.0H)
z, 1.7 Hz), 4.72 (1H, d, J = 7.9 Hz), 4.
74 (1H, d, J = 7.9 Hz), 5.34 (1H, dd, J
= 10.0Hz, 3.0Hz), 5.37 (1H, dd, J = 10.0
Hz, 7.9 Hz), 5.64 (1H, dd, J = 10.0Hz,
7.9Hz), 5.67 (1H, m), 5.69 (1H, t, J = 1
0.0Hz), 7.1-8.0 (35H). (vi) Synthesis of Compound 12-8 Compound 12-7 (0.9 g) was dissolved in 18 ml of ethyl acetate and catalytically reduced at room temperature and atmospheric pressure for 20 hours using platinum oxide as a catalyst.
After removing the catalyst by filtration and neutralizing with 28% sodium methylate (p
H6) The solvent was distilled off. The remaining solid was dissolved in benzene-methanol (1: 1, 24 ml) and 28% sodium methylate was added.
0.6 ml was added and stirred at room temperature for 20 hours.

3.1 ml of 1N HCl was added to the reaction solution, the solvent was distilled off, and column chromatography (“CHP-
20 ", 22 mm x 40 cm, 0-50% acetonitrile) to give a powder of compound 12-8 (0.402 g, 80%).
A portion of this was recrystallized from 0.5N NaOH-ethanol to give a disodium salt of compound 12-8 as an analytical sample.

Mp: 195-199 ° C. (dec). The elemental analysis values as C ₃₆ H ₆₉ O ₁₇ PNa ₂ .4H ₂ O are calculated values: C, 46.85; H, 8.40; P, 3.36; Na,
Measured values for 4.98 C, 47.06; H, 8.32; P, 3.31; N
It was a, 4.83.

[Α] _D −4.5 ° (c 0.8, H ₂ O). ¹ H-NMR (D ₂ O): 0.91 (3 H, bs), 1.33 (3
0H, bs), 1.60 (2H, bs), 3.3-4.5 (28
H, bm). (p) Synthesis of compound 15-2 (FIG. 20) (i) Synthesis of compound 14-1 Mannose peracetate (20.0 g, 51.24 mmol) and 2- [2- (2-chloroethoxy) ethoxy] ethanol (11.232 g) ) Of methylene chloride (300 ml) was added with a solution of boron trifluoride ether complex (25.21 ml) in methylene chloride (25 ml) under ice cooling, and the mixture was stirred overnight at room temperature.

The obtained solution was added to ice water, and 200 ml of chloroform was added for extraction. The organic layer was washed 4 times with water and dried over anhydrous sodium sulfate. The solvent was concentrated under reduced pressure, and the residue was separated by 1700 ml silica gel column chromatography (hexane: ethyl acetate = 2: 1-1: 1) to obtain the desired product. 6.861 g.

[Α] _D ²³ = + 33.3 ° (c 1.35, CHCl ₃ ). ¹ H-NMR (CDCl ₃ , δ): 1.987, 2.039, 2,10
0.2.153 (4s, 3H), 3.397 (t, 2H, J = 5.1
Hz), 3.61-3.85 (m, 10H), 4.064 (m, 1
H), 4.107 (dd, 1H, J = 2.4 Hz, 12.2H
z), 4.286 (dd, 1H, J = 5.1 Hz), 4.873
(D, 1H, J = 1.7 Hz), 5.270 (dd, 1H, J
= 3.4 Hz), 5.288 (t, 1H, J = 10.0 Hz), 5.
364 (dd, 1H). (ii) Synthesis of Compound 14-2 Compound 14-1 (5.861 g) and sodium azide (1.
DMF (50 ml) was added to 146 g) and the mixture was heated with stirring at 60 ° C. for 17 hours.

100 ml of water was added to the obtained solution, and the mixture was extracted with ethyl acetate. The organic layer was washed 3 times with water and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure, and the residue was separated by 350 ml silica gel column chromatography (hexane: ethyl acetate = 1: 1) to obtain the desired product. 4.468 g.

[Α] _D ²¹ = + 35.6 ° (c 1.04, CHCl ₃ ) ¹ H-NMR (CDCl ₃ , δ): 1.988, 2.040, 2.100
, 2.153 (4s, 3H), 3.640 (t, 2H, J = 5.9
Hz), 3.64-3.84 (m, 10H), 4.068 (m, 1)
H), 4.117 (dd, 1H, J = 2.4 Hz, 12.2H
z), 4.287 (dd, 1H, J = 5.1 Hz), 4.874
(D, 1H, J = 1.5 Hz), 5.267 (dd, 1H, J
= 3.7 Hz), 5.289 (t, 1H, J = 10.0 Hz), 5.
362 (dd, 1H). (iii) Synthesis of Compound 14-3 A solution of Compound 14-2 (4.4676 g) in methanol (100 ml) was added with a sodium methylate methanol solution (5 mol / l).
10 drops of was added and stirred at room temperature for 3 hours. To the resulting solution was added an acidic ion exchange resin "Dowex 50W x 8" for neutralization, the resin was filtered off, and the solvent was distilled off under reduced pressure. The residue was dissolved in 100 ml of pyridine and trityl chloride (3.20
3 g) was added and the mixture was heated with stirring at 50 ° C. for 3 hours. Further, trityl chloride (1.231 g) was added, and the mixture was heated at 70 ° C for 14 hours. The resulting solution was cooled to room temperature, benzoyl chloride (3.692 ml) was added, and the mixture was stirred overnight at room temperature. The solution was added to ice water and extracted with 200 ml of chloroform. The organic layer was washed twice with 2N hydrochloric acid and three times with water, and dried over anhydrous sodium sulfate. The solvent was concentrated under reduced pressure, 200 mg of paratoluenesulfonic acid was added to the residue, and 50 ml of a mixed solvent of chloroform: methanol: water = 65: 15: 1 was added and dissolved.
The mixture was heated and stirred at 50 ° C for 5 hours.

The obtained solution was neutralized with triethylamine, and the solvent was evaporated under reduced pressure. 50 ml of water was added to the residue and extracted with chloroform. The solvent was distilled off under reduced pressure, and the residue was
Separation was performed by 500 ml of silica gel column chromatography (hexane: ethyl acetate = 2: 1-1: 1) to obtain the desired product. 2.4106g, 42.0%.

[Α] _D ²³ = −107.2 ° (c 1.75, CHCl ₃ ). ¹ H-NMR (CDCl ₃ , δ): 3.376 (t, 2H,
J = 5.1 Hz), 3.68-3.86 (m, 12H), 3.92-3.96
(M, 1H), 4.166 (brd, dd), 4.196 (br
d, 1H), 5.168 (d, 1H, J = 1.5 Hz), 5.70
2 (dd, 1H, J = 3.4 Hz), 5.823 (t, 1H,
J = 10.0Hz), 5.993 (dd, 1H), 7.23-7.63
(M, 9H), 7.80-8.12 (m, 6H). (iv) Synthesis of compound 14-4 To a solution of compound 14-3 (2.4066 g) in pyridine (20 ml) was added dropwise a solution of diphenylphosphoric acid chloride (1.493 g) in pyridine (3 ml) under ice cooling, and the mixture was stirred overnight at room temperature. . The resulting solution was added to ice water and extracted with chloroform (100 ml). The organic layer was washed twice with 2N hydrochloric acid and three times with saturated brine, and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure, and the residue was separated by 500 ml silica gel column chromatography (hexane: ethyl acetate = 2: 1) to obtain the desired product. 2.848 g, 87.2%.

[Α] _D ²⁷ = −58.6 ° (c 1.04, CHCl ₃ ). ¹ H-NMR (CDCl ₃ , δ): 3.384 (t, 2H,
J = 5.0 Hz), 3.68-3.75 (m, 9H), 3.83-3.89
(M, 1H), 4.39-4.43 (m, 1H), 4.45-4.48
(M, 2H), 5.092 (d, 1H, J = 1.7 Hz), 5.
683 (dd, 1H, J = 2.5 Hz), 5.86-5.92 (m,
2H), 7.10-7.59 (m, 19H), 7.80-8.08 (m, 6
H). (v) Synthesis of Compound 14-5 Compound 14-4 (2.8443 g) and paratoluenesulfonic acid monohydrate (613.5 mg) were dissolved in a mixed solvent of 20 ml of methanol and 20 ml of ethyl acetate to prepare a Lindlar catalyst (1.0
g) was added, and the mixture was stirred under a hydrogen atmosphere of 50 psi for 6 hours. A Lindlar catalyst (1.0 g) was further added, and the mixture was stirred under a hydrogen atmosphere of 50 psi for 3 hours.

The catalyst was filtered and the solvent was distilled off under reduced pressure to obtain the desired product. 3.075 g. This compound was used for the next step synthesis without any particular purification.

(Vi) Synthesis of Compound 15-1 Compound 14-5 (2.1047), 2-palmitylstearic acid (1.354 g), N-hydroxysuccinimide (306 mg)
And a mixture of triethylamine (0.683 ml) are dissolved in 40 ml of methylene chloride and 20 ml of hexane and D is added to this solution.
CC (549 mg) was added, and the mixture was stirred overnight at room temperature. The solution was added to ice water and extracted with chloroform. The organic layer was washed with water and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was separated by 200 ml silica gel column chromatography (hexane: ethyl acetate = 2: 1-1:
1), the desired product was obtained. 796 mg.

[Α] _D ²⁷ = -52.4 (c 1.11, CHCl ₃ ). ¹ H-NMR (CDCl ₃ , δ): 0.876 (t, 6H,
J = 6.9 Hz), 1.18-1.39 (m, 56H), 1.52-1.60
(M, 4H), 1.95-2.01 (m, 1H), 3.43-3.47
(M, 2H), 3.538 (m, 2H), 3.62-3.77 (m, 11
H), 3.85-3.89 (m, 1H), 4.39-4.48 (m, 3
H), 5.103 (d, 1H, J = 1.8 Hz), 5.679 (d
d, 1H, J = 2.7 Hz), 5.86-5.93 (m, 2H),
6.062 (brt, 1H), 7.09-7.59 (m, 19H), 7.
80-8.08 (m, 6H). (vii) Synthesis of compound 15-2 To compound 15-1 (780 mg) and platinum oxide (50 mg) were added ethyl acetate (20 ml) and methanol (10 ml), and the mixture was stirred overnight under a hydrogen atmosphere at normal pressure. The catalyst was filtered and the solvent was distilled off under reduced pressure. To the residue, 2 ml of benzene and 6 ml of methanol were added and dissolved, and a sodium methylate methanol solution (5 mol /
The pH was adjusted to 11 by adding 30 drops of l), and the mixture was stirred overnight at room temperature.

After the solution was neutralized with 1N hydrochloric acid, the solvent was distilled off under reduced pressure. The residue was dissolved in 2 ml of a mixed solvent of chloroform: methanol: water = 65: 15: 1 and subjected to 150 ml of silica gel column chromatography (chloroform: methanol: water = gradient elution from 65: 25: 3 to 60: 35: 7, The total amount was 2000 ml and 100 fractions) were separated. Fractions 38-47 were concentrated. The residue was dissolved in a mixed solvent of chloroform: methanol: water = 65: 15: 1, treated with a strongly acidic ion exchange resin “Dowex 50W × 8”, and then the resin was filtered off. The solvent is concentrated under reduced pressure,
Dissolved in a mixed solvent of chloroform: methanol = 9: 1, "Sephadex LH-20" (22mmφ x 400mm
, Chloroform: methanol = 9: 1 elution). Fractions 5-10 were concentrated to obtain the desired product.
230 mg, 65.8%.

[Α]_D ^{twenty three}= + 15.3 ° (c 1.12, chloro
Form: methanol = 9: 1). (q) Synthesis of compound 28-2 (Fig. 21) (i) Synthesis of Compound 11-4 To 0.745 g of Compound 11-3, 4 ml of thionyl chloride was added, and 5
The mixture was refluxed under heating for an hour. Thionyl chloride was distilled off under reduced pressure,
Benzene was added to the residue and evaporated under reduced pressure (twice). It
It was used in the following reaction without the above purification.

(Ii) Synthesis of Compound 28-1 To 1.175 g of Compound 27-4, 10 ml of methylene chloride was added and dissolved, and the mixture was stirred under ice cooling. Triethylamine 350 μ
1 was added, and the whole amount of compound 11-4 obtained in (a) was dissolved in 5 ml of methylene chloride and added, and the temperature was raised to room temperature.
Stir for hours.

The solvent was distilled off under reduced pressure, and the residue was purified by silica gel chromatography (eluting solvent: n-hexane-
Ethyl acetate 1: 1), 1.23 as the desired product as colorless amorphous
2 g was obtained.

¹ H-NMR (δ, CDCl ₃ ): 0.88
(T, 6H, J = 7.0 Hz), 1.18-1.31 (m, 56
H), 1.33-1.41 (m, 2H), 1.52-1.60 (m, 2
H), 1.93-1.99 (m, 1H), 3.33-3.34 (m, 2)
H), 3.37-3.42 (m, 4H), 3.45-3.48 (m, 2)
H), 3.56-3.64 (m, 2H), 3.73 (ddd, 1H,
J = 3.6 Hz, 7.2 Hz, 10.9 Hz), 3.94 (ddd,
1H, J = 3.5 Hz, 4.7 Hz, 11.2 Hz), 4.06-4.
10 (m, 1H), 4.41 (ddd, 1H, J = 6.3 Hz,
11.5Hz, 8.7Hz), 4.47 (ddd, 1H, J = 2.5
Hz, 7.3 Hz, 11.5 Hz), 4.89 (d, 1H, J = 8.
0 Hz), 5.45 (dd, 1H, J = 8.0 Hz, 9.8 H
z), 5.45 (t, J = 9.8 Hz), 5.85 (t, 1H, J
= 9.8 Hz), 5.96 (brs, 1H), 7.13-7.54
(M, 19H), 7.81 (dd, 2H, J = 1.2 Hz, 8.3
Hz), 7.90 (dd, 2H, J = 1.2 Hz, 8.3 H
z), 7.95 (dd, 2H, J = 1.2 Hz, 8.3 Hz). [Α] _D ²⁵ = + 1.3 ° (c = 1.02, CHCl ₃ -MeO
H 1: 1) _Rf : 0.27 (n-hexane-AcOEt 2: 3). (iii) Synthesis of compound 28-2 To 1.216 g of compound 28-1, 1.2 ml of tetrahydrofuran was added and dissolved with 30 ml of tetrahydrofuran and 10 ml of methanol. Platinum oxide 0.01
5 g was added, and catalytic reduction was carried out under normal pressure for 6 days. The catalyst was filtered off, and the solvent was distilled off under reduced pressure. The residue was dissolved by adding 8 ml of benzene and 8 ml of methanol, 22 drops of 28% sodium methoxide methanol solution (pH = 11) was added, and the mixture was stirred at room temperature for 11.5 hours.

The mixture was ice-cooled and neutralized with 1N hydrochloric acid, and the solvent was distilled off under reduced pressure. The residue was purified by "Sepphadex LH-20" column (resin: about 150 ml, elution solvent: chloroform-methanol-water 10: 10: 3). 0.640 g of the target compound was obtained as a colorless powder.

¹ H-NMR (δ, CDCl ₃ -D ₂ O
5: 1): 0.88 (t, 6H, J = 7.0 Hz), 1.16-1.
34 (m, 56H), 1.34-1.44 (m, 2H), 1.46-1.59
(M, 2H), 2.04-2.11 (m, 1H), 3.26-6.36
(M, 19H). ¹³ C-NMR (δ, CDCl ₃ -CD ₃ OD-D ₂ O
10: 10: 3): 14.25, 23.04, 27.83, 29.71, 29.84
, 30.02, 32.30, 33.07, 39.23, 47.78, 63.09
(J = 2.4 Hz), 69.00-70.50, 69.28, 74.12,
75.70, 76.42 (J = 5.5 Hz), 103.55, 178.38. [Α] _D ²⁶ = -14.2 ° (c = 1.05, CHCl ₃ -MeO
H 1: 1). _{R f: 0.44 (CHCl 3 -MeOH} -H 2 O 60:35:
7). FAB-MS: [M + H] ⁺ ; M / Z = 882, [M + N
a] ⁺ ; M / Z = 904. (r) Synthesis of compound 18-2 (FIG. 22) (i) Synthesis of compound 17-1 Galactose peracetate (10.0 g) and 2- [2
-(Chloroethoxy) ethoxy] -ethanol (5.616
g) in methylene chloride (150 ml) solution of boron trifluoride ether complex (12.6 ml) in methylene chloride (30 ml)
The solution was added under ice cooling and stirred overnight at room temperature.

The obtained solution was added to ice water, and chloroform (150 ml) was added for extraction. Wash the organic layer twice with water,
It was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was separated by 1000 ml silica gel column chromatography (hexane: ethyl acetate = 2: 1-1:
1), the desired product was obtained. 6.51 g.

[Α] _D ²⁰ = −0.4 ° (c 1.098, chloroform). ¹ H-NMR (CDCl ₃ , δ): 1.986, 2.051, 2.0
63, 2.152 (4s, 3H), 3.63-3.78 (m, 11
H), 3.95-3.98 (m, 1H), 3.917 (brt, 1
H), 4.131 (dd, 1H, J = 6.8 Hz, 11.2H
z), 4.178 (dd, 1H, J = 6.6 Hz), 4.576
(D, 1H, J = 8.1 Hz), 5.023 (dd, 1H, J
= 3.4 Hz), 5.212 (dd, 1H, J = 10.5Hz),
5.390 (brd, 1H). (ii) Synthesis of Compound 17-2 Compound 17-1 (6.445 g) and sodium azide (1.
DMF (50 ml) was added to 26 g) and the mixture was heated with stirring at 60 ° C. for 17 hours.

Water (100 ml) was added to the obtained solution, and the mixture was extracted with ethyl acetate. The organic layer was washed with water and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure, and the residue was separated by 500 ml silica gel column chromatography (hexane: ethyl acetate = 2: 1.5-1: 1) to obtain the desired product. 5.30g.

[Α] _D ¹⁷ = −3.2 ° (c 1.04, CHCl ₃ ). ¹ H-NMR (CDCl ₃ , δ): 1.984, 2.048, 2.0
60, 2.148 (4s, 3H), 3.398 (t, 2H, J =
5.0 Hz), 3.63-3.69 (m, 8H), 3.73-3.78
(M, 1H), 3.95-3.98 (m, 1H), 3.910 (br
t, 1H), 4.131 (dd, 1H, J = 6.8 Hz, 1
1.2Hz), 4.176 (dd, 1H, J = 6.3Hz), 4,5
71 (d, 1H, J = 7.8 Hz), 5.023 (dd, 1
H, J = 3.4 Hz), 5.210 (dd, 1H, J = 10.5H
z), 5.387 (dd, 1H, J = 1.0 Hz). (iii) Synthesis of compound 17-3 To a solution of compound 17-2 (4.957 g) in methanol (70 ml) was added sodium methylate methanol solution (5 mol / l).
10 drops were added, and the mixture was stirred at room temperature for 3 hours. The solution was neutralized by adding an acidic ion exchange resin “Dowex 50W × 8”, and the solvent was evaporated under reduced pressure. The residue was dissolved in pyridine (30 ml), and diphenylphosphoric chloride (3.
A pyridine solution of 688 g) was added dropwise, and the mixture was stirred at room temperature for 8 hours. Benzoyl chloride (4.55 ml) was added to the resulting solution, and the mixture was stirred overnight.

The solution was added to ice water, and chloroform (200 m
l). The organic layer was washed twice with 2N hydrochloric acid and three times with saturated brine, and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure, and the residue was separated by 500 ml silica gel column chromatography (hexane: ethyl acetate =
2: 1-1: 1), the target product was obtained. 3.468 g.

[Α] _D ²¹ = + 91.8 ° (c 1.02, CHCl ₃ ). ¹ H-NMR (CDCl ₃ , δ): 3.305 (t, 2H,
J = 5.0 Hz), 3.35-3.63 (m, 8H), 3.753
(M, 1H), 3.978 (m, 1H), 4.196 (brt,
1H), 4.364 (ddd, 1H, J = 5.6 Hz, 10.7H
z, 8.3 Hz), 4.467 (ddd, 1H, J = 7.1 H
z, 9.0 Hz), 4.859 (d, 1H, J = 8.1 Hz),
5.524 (dd, 1H, J = 3.4 Hz), 5.746 (dd,
1H, J = 10.5Hz), 5.881 (brd, 1H), 7.14
-7.62 (m, 19H), 7.78-8.05 (m, 6H). (iv) Synthesis of compound 17-4 Compound 17-3 (3.424 g) and paratoluenesulfonic acid monohydrate (739 mg) were dissolved in a mixed solvent of methanol (20 ml) and ethyl acetate (120 ml) to give a Lindlar catalyst ( 1.2 g) was added, and the mixture was stirred under a hydrogen atmosphere of 50 psi for 7 hours.

The catalyst was filtered and the solvent was distilled off under reduced pressure to obtain the desired product (4.00 g). This compound was used for the next step synthesis without any particular purification.

(V) Synthesis of Compound 18-1 Compound 17-4 (2.00 g), 2-palmitylstearic acid (1.188 g), N-hydroxysuccinimide (269 mg)
And 4-dimethylaminopyridine (523 mg) in DMF
N, N'-Dicyclohexylcarbodiimide (482 mg) was added to the (100 ml) solution, and the mixture was stirred overnight at room temperature. N, N '
-Dicyclohexylcarbodiimide (200 mg) was further added, and the mixture was further stirred overnight.

The N, N'-dicyclohexylurea that had come out was filtered off, and the solvent was distilled off at 40 ° C. Ethyl acetate (40 ml) was added to the residue, and the insoluble material was filtered off. The filtrate was concentrated under reduced pressure and the residue was separated by 200 ml silica gel column chromatography (hexane: ethyl acetate = 2: 1-1:
1), the desired product was obtained. 773 mg.

[Α] _D ²¹ = + 58.9 ° (c 1.13, CHCl ₃ ). ¹ H-NMR (CDCl ₃ , δ): 0.877 (t, 6H,
J = 6.9 Hz), 1.16-1.34 (m, 56H), 1.52-1.61
(M, 4H), 1.91-1.99 (m, 1H), 3.30-3.33
(M, 2H), 3.35-3.63 (m, 8H), 3.73-3.77
(M, 1H), 3.97-4.01 (m, 1H), 4.196 (br
t, 1H), 4.357 (ddd, 1H, J = 5.6 Hz,
10.7Hz, 8.5Hz), 4.468 (ddd, 1H, J = 7.
1 Hz, 9.0 Hz), 4.848 (d, 1H, J = 8.1 H
z), 5.523 (dd, 1H, J = 3.5 Hz), 5.747
(Dd, 1H, J = 10.4Hz), 5.878 (brd, 1
H), 7.13-7.62 (m, 19H), 7.77-8.05 (m, 6
H). (vi) Synthesis of Compound 18-2 Tetrahydrofuran (20 ml) and methanol (10 ml) were added to Compound 18-1 (506 mg) and platinum oxide (50 mg), and the mixture was stirred overnight under a hydrogen atmosphere at normal pressure. Filter the catalyst,
The solvent was distilled off under reduced pressure. Methanol (15 ml) was added to the residue to dissolve it, and 20 drops of sodium methylate methanol solution (5 mol / l) was added to adjust the pH to 11, and the mixture was stirred at room temperature for 5 hours.

After the solution was neutralized with 1N hydrochloric acid, the solvent was distilled off under reduced pressure. The residue is chloroform: methanol =
It was dissolved in a 1: 1 mixed solvent, 2.5 g of silica gel was added, and the solvent was evaporated under reduced pressure. The dried silica gel obtained
150 ml of chloroform: methanol: water = 65: 25: 4
The mixture was packed in a silica gel column packed with the solvent mixture described above, and the fraction containing the desired product was collected (22 mmφ × 400 mm, chloroform: methanol: water = 65: 25: 4 to 2 :).
2: Gradient elution up to 0.5, total volume 1000 ml, 100 fractions). Fractions 13-41 were concentrated. The residue was dissolved in a mixed solvent of chloroform: methanol = 9: 1 and filled with the same solvent "Sephadex LH-20".
(22mmφ × 400mm, chloroform: methanol = 9:
(1 elution). Fractions 5-8 were concentrated to give the desired product. 201 mg.

[Α] _D ²⁸ = −3.8 ° (c 1.09, chloroform: methanol = 9: 1). Mass M / Z: 882 (M + H). ¹ H-NMR (CDCl ₃ -CD ₃ OD = 5/1,
δ): 0.884 (t, 6H), 1.20-1.33 (m, 56H),
1.36-1.44 (m, 2H), 1.51-1.59 (m, 2H), 2.
05-2.11 (m, 1H), 3.416 (brt, 2H, J =
5.0 Hz), 3.51-3.77 (m, 12H), 3.931 (br
d, 1H, J = 1.0 Hz), 4.01-4.20 (m, 3H),
4.267 (d, 1H, J = 7.6 Hz). ¹³ C-NMR (CDCl ₃ -CD ₃ OD = 5/1,
δ): 14.16, 22.87, 27.83, 32.13, 33.18, 39.1
8, 47.88, 64.55 (J = 4.9 Hz), 67.97, 68.53
, 70.17, 70.31, 70.51, 70.60, 71.51, 73.33
, 73.54 (J = 8.1 Hz), 103.77, 177.71. Example 3 (Preparation of liposome) Compound of the compounds of the present invention synthesized in Examples 1 and 2
Liposomes were prepared as follows using each of 16 kinds of compounds except 1205 and 28-2.

80 μmol of L-α-dipalmitoylphosphatedylcholine, 80 μmol of cholesterol, 8 μmol of dicetylphosphoric acid and 16 μmol of the compound of the present invention were dissolved in a filtrate of chloroform and methanol (volume ratio 1: 1).
Next, the organic solvent was removed in a nitrogen gas stream to form a lipid film on the glass wall of the centrifuge tube.

To this, 8 ml of 1 mM inulin phosphate-buffered saline (pH 7.4) preheated to about 45 ° C. was added, shaken, and further sonicated gently to prepare a liposome suspension. . This was heated to 45 to 60 ° C. and then passed through a polycarbonate membrane filter having a pore size of 0.08 μm to prepare a suspension of liposomes having a particle size of about 0.08 μm.

Compounds 1205 and 28-2 can also be made into liposomes in the same manner.

Similarly, a liposome suspension was prepared by using the four control compounds synthesized in Example 1 instead of the compound of the present invention.

Example 4 (Evaluation test) a. Samples 4 compounds of the invention synthesized in Example 1 and 4 control compounds were used instead of 1 mM inulin ³
A total of 8 types of liposomes were obtained in the same manner as in Example 3 except that 1 mM inulin containing 140 μCi of H-inulin was used, and these were used as samples.

B. Test method Each of the prepared 8 types of samples was used for SD male rats (body weight
200-250 g) L-α per 100 g body weight from hindlimb vein
-Dipalmitoylphosphatidylcholine and cholesterol were infused at 5 μmol.

Six hours after the administration, the rat was sacrificed, about 200 mg of each tissue was taken, dried and burned in a burning device, and its radioactivity was determined by the liquid scintillation method.
The inulin concentration per gram was determined.

C. Results Results are shown in Figures 23-26.

From these results, liposomes containing the lipid derivative having at least two alkyl groups (the present invention)
In comparison with the liposome (control) produced by the same method as the method of the present invention except that a compound having one alkyl group is used, the organ recognition is improved.

[0278]

EFFECTS OF THE INVENTION According to the present invention, liposomes having excellent stability, organ directivity, and drug retaining function are easily provided.

[Brief description of drawings]

1 shows the reaction in Example 1. FIG.

2 shows the reaction in Example 1. FIG.

FIG. 3 shows the reaction in Example 1.

FIG. 4 shows the reaction in Example 1.

FIG. 5 shows the reaction in Example 1.

FIG. 6 shows the reaction in Example 1.

FIG. 7 shows the reaction in Example 1.

FIG. 8 shows the reaction in Example 1.

FIG. 9 shows the reaction in Example 2.

FIG. 10 shows the reaction in Example 2.

FIG. 11 shows the reaction in Example 2.

FIG. 12 shows the reaction in Example 2.

FIG. 13 shows the reaction in Example 2.

FIG. 14 shows the reaction in Example 2.

FIG. 15 shows the reaction in Example 2.

16 shows the reaction in Example 2. FIG.

FIG. 17 shows the reaction in Example 2.

FIG. 18 shows the reaction in Example 2.

FIG. 19 shows the reaction in Example 2.

FIG. 20 shows the reaction in Example 2.

FIG. 21 shows the reaction in Example 2.

FIG. 22 shows the reaction in Example 2.

FIG. 23 shows the results of Example 4.

FIG. 24 shows the results of Example 4.

FIG. 25 shows the results of Example 4.

FIG. 26 shows the results of Example 4.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Shiro Miyoshi 206-1 Anku, Mishima City, Shizuoka Prefecture Tamura Heights No. 5 (72) Inventor Ao Masaru 2-529-4 Asahimotocho, Nara City, Nara Prefecture Excel Heights B-308 (72) Inventor Hitoshi Yamauchi 2-32-12 Okuto, Katsushika-ku, Tokyo (72) Inventor Naoichi Murahashi 7-2-4 Matsumaedai Moriya-cho, Kitasoma-gun, Ibaraki Prefecture (72) Inventor Atsushi Sasaki Tsukuba, Ibaraki Prefecture Kasuga 4-19-13 Eisai Shiyama Dormitory 307 (72) Inventor Hiroshi Watanabe 7-131 Shinmatsudo, Matsudo City, Chiba Prefecture 7-131 Sevenpia 301 (72) Inventor Hideo Kaneko 1-1-25 Nakamuracho, Minami-ku, Yokohama City, Kanagawa Prefecture

Claims (4)

[Claims] 1. At least polar lipid 1 as a raw material
Mol, compound giving positive or negative charge 0.05 to 0.5 mol, cholesterol 0.3 to 1.5 mol, degree of polymerization of 3 in molecule
6 polyethylene glycol and at least 2
A method for producing liposomes, which comprises using 0.02 to 0.5 mol of a compound having an alkyl group having 5 to 20 carbon atoms and 50 to 100 l of an aqueous solvent in this proportion. 2. A polar lipid 1 mol, a compound giving a positive or negative charge 0.05 to 0.5 mol, cholesterol 0.3 to 1.
The solvent is removed from an organic solvent containing 5 moles and 0.02 to 0.5 moles of a polyethylene glycol having a degree of polymerization of 3 to 6 and at least two compounds having an alkyl group having 5 to 20 carbon atoms in this ratio. A lipid film is formed, and then an aqueous solution of the drug to be incorporated into the liposome is added to the lipid film in an amount of 50 to 100 l per mol of the polar lipid to form a liposome.
The liposome of claim 1 is selectively collected.
A method for producing the described liposome. 3. A polar lipid 1 mol, a compound giving a positive or negative charge 0.05 to 0.5 mol, cholesterol 0.3 to 1.
Drugs to be incorporated into liposomes in an organic solvent containing 5 mol and 0.02 to 0.5 mol of a polyethylene glycol having a degree of polymerization of 3 to 6 and at least two compounds having an alkyl group having 5 to 20 carbon atoms in this ratio in 5 mol. 50-100 liters of the aqueous solution of (1) are added to 1 mol of polar lipid, and then the mixed solution is sonicated to form a w / o type emulsion, the organic solvent is removed under reduced pressure, and vortexing is further performed. The method for producing liposomes according to claim 1, wherein the organic solvent is again removed under reduced pressure after forming the / w emulsion. 4. At least polar lipid 1 as a raw material
Mol, a compound giving positive or negative charge 0.05 to 0.5 mol, cholesterol 0.3 to 1.5 mol, polyethylene glycol having a degree of polymerization of 3 to 6 and a compound having at least two alkyl groups having 5 to 20 carbon atoms in the molecule 0.02 ~ 0.
A method for producing liposomes, characterized in that 5 mol is contained in this ratio, and 50 to 100 l of an aqueous solvent is used per mol of polar lipid as well as an organic solvent containing a drug.