JP2854203B2 - Method for producing liposomes - Google Patents

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-carrying liposome as a so-called drug delivery system, and more particularly, to a method for producing a liposome which can easily carry an organ-directed sensor.

[0002]

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

In order to solve this problem, attempts have been made to use liposomes as organ-directing sensors such as sugars and antibodies. That is, the liposome is coated with the lipid derivative of the organ directional sensor, and the organ directional sensor is exposed on the surface 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,
The lipid derivative of the organ-directed sensor does not efficiently cover the liposome, and in some cases, the particle size of the liposome is not constant, and in other cases, the stability of the liposome is low. Was.

[0004]

The object of the present invention is to provide a liposome which is excellent in stability, excellent in avoiding capillary endothelial tissue, excellent in directing to a specific organ, and excellent in drug retention function. Aim.

[0005]

Means for Solving the Problems The present inventors have made intensive studies to solve the above-mentioned problems, and as a result, have accomplished the present invention.

Hereinafter, the present invention will be described sequentially.

[0007] The present invention provides at least
1 mole of polar lipid, compound giving positive or negative charge
05 to 0.5 mol, cholesterol 0.3-1.5 mol, organ directional sensor in the molecule, polyethylene glycol having a degree of polymerization of 3-6 and Compound 0.02 mole and an aqueous solvent having at least two alkyl groups having a carbon number of 5-20 50-10
The present invention relates to a method for producing liposomes, wherein 0 l is used in this ratio.

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

Examples of the compound providing a positive charge include:
Examples of the compound that gives an aliphatic amine such as stearylamine and a negative charge include dicetyl phosphoric acid. These compounds are used in an amount of 0.05 mol per mol of polar lipid.
Used in proportions of .about.0.5 mol.

Cholesterol is used as a stabilizer. If the amount of cholesterol is too large, liposome formation is inhibited, or even if liposomes are formed, they are physicochemically unstable. Even stable liposomes cannot be made. Therefore, 0.3 to 1.5 per mole of polar lipid
Preference is given to using them in molar proportions.

Organ directional sensor in molecule , degree of polymerization 3 to
The polyethylene glycol 6 and a compound having at least two alkyl groups having 5 to 20 carbon atoms (hereinafter, sometimes referred to as a “lipid derivative”) cover the liposome and serve as an organ-directing sensor.

As the polyethylene glycol, those having a degree of polymerization of ethylene glycol of 3 to 6 must be used. Desirable results cannot be obtained by using a polymer having a degree of polymerization other than this.

The compounds having at least two alkyl groups of carbon <br/> prime 5-20 in the molecule of the lipid derivative and polyethylene
To explain with respect to the binding mode of glycol, in the lipid derivative molecule, and if the compound having a functional group such as carboxyl group polyethylene glycol may engage directly stitched to the hydroxyl group is bonded, it may engage binding to the hydroxyl group In some cases, the compound is bonded to a linker having a functional group, for example, glutamic acid. The latter is advantageous because two compounds can be attached to the linker. Ma
In addition, the hydroxyl group of polyethylene glycol is an amino group or
Also replaces other reactive functional groups,
As described above, the alkyl group is directly bonded to the reactive functional group.
And when linked via a linker
There is.

[0014] donor alkyl group include a fatty acid, fatty alcohols and aliphatic primary amines. These fatty acids, aliphatic alcohols, aliphatic amines, and the like may be any of a linear type and a branched type.

An alkyl group having 5 to 20 carbon atoms has at least 2
Compounds that individual chromatic include ethylene glycol, rather good even like alkyl <br/> Le group having 2 or more carbon atoms 5-20 via a linker glutamic acid is connected, also two or more
A branch having 5 to 20 carbon atoms
Those having a alkyl group may be used.

[0016] The lipid derivative of the present invention should contain at least
The number of carbon atoms in the alkyl group of the compound having two alkyl groups is 5 in both the linear and branched alkyl groups.
-20, preferably 12-18 .

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 (ie, not functionalized).

The hydroxyl group at one end of polyethylene glycol of the lipid derivative is bonded directly to the alkyl group functional group or via a linker as it is or as described above, by substituting with another functional group such as an amino group. , The other end is an antibody,
Ligands such as antigens, lectins, cell adhesion factors; proteins such as cell adhesion factors, antibodies, and immunoglobulins; sugars; amino acids; nucleic acids and nucleic acid-based compounds; .

The ratio of the lipid derivative to the polar lipid greatly affects the quality of the obtained liposome. That is,
When the amount is too large, a stable liposome cannot be produced, and when the amount is too small, the resulting liposome membrane does not change at all, and the effect of blending the lipid derivative is not exhibited. For this reason, lipid derivatives are used per mole of polar lipid.
It is used in a proportion of 0.02 to 0.5 mol, preferably 0.1 to 0.5 mol.

The drug to be included in the liposome includes, for example, 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 drugs are usually used by dissolving them in an aqueous solvent, but they may be used by being added to an organic solvent.

The concentration of these drugs in an aqueous solvent is not particularly limited, and is appropriately determined depending on the method of use, use, and the like of the liposome. The aqueous solvent is desirably 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 of preparing the liposome is the same as when the drug is added to the aqueous solvent.

The aqueous solution of the drug is used per mole of polar lipid.
Used in a proportion of 50-100 l.

The process itself for producing drug-carrying liposomes using an aqueous solution of a polar lipid, a compound imparting a positive or negative charge, cholesterol, a lipid derivative and a drug in the above-mentioned ratio as a raw material is performed by a special method. No need to be. For example, Nojima et al., “Liposomes” (Nan-Edo 1989
Published on September 15, 2008), which describes a number of methods for preparing liposomes, and any of these methods can be used in the present invention. As a method for preparing multilamellar liposomes, the method of Bangham (ADBangham et al., J. Mol. Biol., 13 ,
238 (1965) is known.In this method, a solvent is removed from an organic solvent solution of liposome raw materials by a stream of nitrogen gas or distillation under reduced pressure to form a lipid film on the container wall, and an aqueous solvent is added thereto. In addition, the lipid film is allowed to stand for a while to hydrate the lipid film, and then exposed to ultrasonic waves or vortexing to prepare liposomes. The resulting multilamellar liposomes have a large particle size and containment (drug)
Has high retention ability. On the other hand, small unilamellar liposomes can be prepared by sonication, ethanol injection, French press, etc., but generally have a low retention volume for water-soluble drugs. 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,
Methods for preparing large unilamellar liposomes, such as a melting method and a reverse phase evaporation method, are preferable because relatively stable liposomes can be obtained and a large amount of a water-soluble drug is retained.

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

The following three examples are shown as more preferable embodiments of the present invention. The first is 1 mol of polar lipid, 0.05 to 0.5 mol of a compound providing a positive or negative charge, 0.3 to 1.5 mol of cholesterol and an organ-directed sensor in the molecule ,
Compound 0.02 with polyethylene glycol and at least two alkyl groups of carbon number 5-20 of polymerization degree 3-6
The solvent was removed from the organic solvent containing 0.5 moles in this proportion to form a lipid film, and the aqueous solution of the drug to be included in the liposome was then added to 50 moles per mole of phospholipid.
100100 l is added to the lipid film to form liposomes, and liposomes having a particle size of 50 to 1000 nm are selectively collected.

Although there is no particular limitation on the organic solvent, those which can dissolve and easily remove polar lipids, cholesterol and lipid derivatives, such as chloroform, ether, etc.
Chloroform-methanol mixture and the like can be mentioned.

There is no particular limitation on the concentration of the organic solvent solution as long as the organic solvent can dissolve the polar lipid.

The formation of the lipid film from the organic solvent solution can be carried out, for example, by removing the organic solvent by a stream of nitrogen gas to form a lipid film on the vessel wall, or by distilling the organic solvent under reduced pressure.

There is no particular limitation on the method for preparing a drug-retaining liposome by adding an aqueous solution of a drug to a lipid film. For example, an aqueous solution is added to a lipid film and the mixture is allowed to stand for a while to hydrate the lipid film. This can then be by exposure to ultrasound or by vortexing.

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

As a second embodiment, 1 mol of polar lipid, 0.05 to 0.5 mol of a compound imparting a positive or negative charge, 0.3 to 1.5 mol of cholesterol and organ finger in the molecule are used.
Tropism sensor, an aqueous solution of the agent to be included in the liposome in an organic solvent containing the compound from 0.02 to 0.5 mole of an alkyl group having polyethylene glycol and at least two carbon number 5-20 of polymerization degree 3-6 at this ratio Was added to 50 mol / l of phospholipid, and the mixed solution was sonicated to form a w / o emulsion, and then the organic solvent was removed under reduced pressure.
This is based on a so-called reverse evaporation method in which the organic solvent is removed again under reduced pressure if necessary after the emulsion is formed.

The purpose of subjecting the mixed solution of the organic solvent solution and the aqueous solution of the drug to sonication is to prepare a w / o emulsion, and therefore, the sonication is carried out under known ordinary conditions. be able to. Regarding the removal of the organic solvent, the present invention does not need to be replaced with a particularly ordinary method.

Vortexing is performed for the purpose of converting a w / o emulsion to an o / w emulsion. When the organic solvent is removed from the obtained o / w emulsion under reduced pressure, a target drug-containing 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, the 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 the liposome produced by the same method, when an organ directional sensor is carried at one end of ethylene glycol, the organ recognition of the liposome is improved, and the accumulation of the liposome in the organ or tissue is enhanced.

[0037]

【Example】

Example 1 (Synthesis Example (1)) 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.
Various compounds (invention) 256, 852, 331 and 531 were synthesized as follows.

The reactions in these synthesis 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)
ol, 1.3 eq) in methylene chloride (150 ml) was added with a solution of boron trifluoride ether complex (12.6 ml, 4.0 eq) in methylene chloride (30 ml) under ice-cooling, and the mixture was stirred overnight at room temperature. The obtained solution was added to ice water, extracted with chloroform (150 ml). The organic bath was washed twice with water and dried over anhydrous sodium sulfate. The solvent is distilled off under reduced pressure,
The residue was separated by 1000 ml of 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 × 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 = 3.4 Hz), 5.212 (dd, 1H, J = 10.5 Hz),
5.390 (brd, 1H).

2) Synthesis of compound 225 DMF (50 ml) was added to compound 254 (3.445 g, 12.91 mmol) and sodium azide (1.26 g, 19.4 mmol, 1.5 eq).
Was added and the mixture was heated and stirred at 60 ° C. for 17 hours. Water (100 ml) was added to the resulting solution and 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 of 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 × 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) were added to methanol (20 ml).
And ethyl acetate (40 ml). To the solution was added a Lindlar catalyst (0.2 g) and the mixture was stirred under a 50 psi hydrogen atmosphere for 8 hours. The catalyst was removed by filtration, and the solvent was distilled off under reduced pressure. The residue was methylene chloride (20 ml) and hexane (10 ml)
And 2-palmityl stearic acid (0.662 g),
N-hydroxysuccinic acid (0.150 g), triethylamine (0.181 ml) and dicyclohexylcarbodiimide
(0.268 g) and stirred overnight. The solution was concentrated under reduced pressure, ethyl acetate was added to the residue, and insolubles were removed by filtration. 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 (c 1.04, chloroform).

[0047] _{^{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), and
Eight drops of M / 1 methanol solution of sodium methylate were added and stirred overnight. Add strongly acidic ion exchange resin "Do
Wex 50w × 8 ”form H was added for neutralization. The solution was filtered, concentrated under reduced pressure, and the residue was treated with Sephadex LH-2.
0 "(chloroform: methanol = 9: 1, 22mmφ × 4
5 cm) to give the desired compound (252 mg, 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 = 7.8 Hz), 8.76 (bt, 1H).

(B) Synthesis of Compound 852 (FIG. 2) A solution of carboxylic acid 828 (180 mg) and N-hydroxysuccinimide (41 mg) in methylene chloride (10 ml) was added with N,
N'-Dicyclohexylcarbodiimide (74 mg) was added, and the mixture was stirred at room temperature for 1 hour. To this solution was added a solution of amino paratoluenesulfonate 818 (190 mg) and triethylamine (90 μl) in methylene chloride (5 ml), and the mixture was stirred at room temperature overnight. After filtering off insolubles, 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 the amide (310 mg, 77 mg).
%).

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

¹ H-NMR (CD ₃ OD): 0.90 (3H, t, J = 7 Hz), 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 = 3.5Hz).

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

Sodium methoxide (55 μl of a 28% methanol solution) was added to a solution of the amide (270 mg) obtained above in methanol (20 ml), and the mixture was stirred at room temperature for 5.5 hours.
After adding “Amberlyst 15E” until the solution becomes neutral, the resin is removed by filtration, the filtrate is concentrated, and the target compound 852 (21
5 mg).

[Α] _D ²⁶ +43.9 c (c 1.0, methanol).

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

¹ H-NMR (CD ₃ OD): 0.90 (3H, t, J = 7 Hz), 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), and BF ₃ .Et ₂ O (5.68 g) was added.
Stirred for days. Dilute the reaction solution with methylene chloride, add water,
% NaHCO ₃ and water, dried, and the solvent was distilled off under reduced pressure. The residue was purified by column chromatography (chloroform) using silica gel (200 g).
-Glycoside compound 327 (4.13 g) 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 The chlorinated 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 distilled off under reduced pressure. Silica gel (150 g)
And purified by 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,
10.0Hz).

3) Synthesis of compound 329 Azide compound 328 (386 mg) and p-toluenesulfonic acid (145 mg) were dissolved in ethanol (20 ml), a Lindlar catalyst (770 mg) was added, and catalytic reduction was carried out at room temperature at 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. 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 material was removed by filtration, 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 using silica gel (60 g) (chloroform-methanol 150: 1). The product was purified again by column chromatography (hexane-ethyl acetate 5: 4) using silica gel (60 g) to obtain the desired 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. Add “Amberlite IRC-
50 "was added, the insolubles were removed by filtration, and the filtrate was concentrated to dryness under reduced pressure. Chloroform was added to the residue, the insoluble material was removed by filtration, and the filtrate was concentrated to dryness under reduced pressure. The residue was washed with hexane under ice-cooling to obtain the desired 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, m
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 material was removed by filtration, 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 using silica gel (60 g) (chloroform-methanol 150: 1). The product was purified again by column chromatography (hexane-ethyl acetate 2: 1) using silica gel (60 g) to obtain the desired 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 stirred at room temperature for 2.5 hours. When the reaction mixture contains "Amberlite IRC
-50 "was added, the insolubles were removed by filtration, and the filtrate was concentrated to dryness under reduced pressure. The residue was added to chloroform, the insolubles were removed by filtration, the filtrate was concentrated to dryness under reduced pressure, and the residue was washed with ether to obtain the desired 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 a control, the above compounds 256, 852, 331
And 531 respectively, four kinds of compounds having one alkyl group having 5 to 20 carbon atoms (control compounds) 22
8, 851, 333 and 529 were synthesized as follows.

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

(A ′) Synthesis of Compound 228 (FIG. 5) 1) Synthesis of Compound 225 β-D-galactose pentaacetate compound 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. Here, 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), it was washed with saturated saline, 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) 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 catalyst were added thereto, and the mixture was catalytically reduced at 50 psi for 4 hours. Add 0.509 g of Lindlar's catalyst and add 50
Catalytic reduction was performed at psi for 6 hours. The catalyst was removed by filtration to obtain 1.001 g of the desired product as a pale brown oil. It was used for the following reaction without further purification.

3) Synthesis of compound 227 To 1.001 g of compound 226, 20 ml of methylene chloride was added, and 214 μl of triethylamine was added thereto to obtain a homogeneous solution, which was stirred under ice-cooling. 0.815 g of (N-palmitoyloxy) succinimide dissolved in 6 ml of methylene chloride was added thereto, 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 = 0.5 Hz, 3.3 Hz), 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 the mixture was stirred under ice cooling. To this was added 6 drops of a 28% sodium methoxide methanol solution to adjust the pH to 12, followed by stirring at room temperature for 12.5 hours. A "Dowex 50X-8" ion exchange resin (H type) was added to neutralize the mixture, and the resin was removed by filtration. The solvent was distilled off under reduced pressure, and the residue was purified by "Sephadex LH-20" (elution solvent: chloroform-methanol 1: 1) to obtain 1.11 g of the desired 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 = 7.6 Hz), 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 N-palmitoyloxysuccinimide (115 mg) in toluene (5 ml) was added to a solution of the amino para-toluenesulfonate 818 (167 mg) in methylene chloride (10 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 = 7 Hz), 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).

To a solution of the palmitoyl compound (141 mg) obtained above in methanol (15 ml) was added sodium methoxide (40 μl of a 28% methanol solution), and the mixture was stirred at room temperature for 5.5 hours. A cation exchange resin "Amberlyst 15E" (manufactured by Rohm and Haas) was added until the solution became neutral, the resin was filtered off, the filtrate was concentrated, and the target compound 851 (1010
4 mg, 90%).

[Α] _D ²⁶ +43.9 c (c 1.0, methanol).

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

¹ H-NMR (CD ₃ OD): 0.90 (3H, t, J = 7 Hz), 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), and triethylamine was dissolved in triethylamine. (65 mg) and stirred at room temperature for 4 hours. The reaction solution was diluted with methylene chloride, washed sequentially with water, 10% aqueous citric acid and water, dried, and the solvent was distilled off under reduced pressure. The residue was purified by column chromatography (chloroform-methanol 100: 1) using silica gel (50 g) to obtain the desired 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 stirred at room temperature for 4 hours. Add “Amberlite IRC-
50 "was added, the insolubles were removed by filtration, and the filtrate was concentrated to dryness under reduced pressure. Chloroform was added to the residue, the insolubles were removed by filtration, the filtrate was concentrated to dryness under reduced pressure, and the residue was washed with ether to obtain the desired 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 solution with methylene chloride, water, 5%
The extract was washed successively with aqueous NaHCO ₃ and water, dried, and the solvent was distilled off 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 hours. The reaction mixture was diluted with ethyl acetate and diluted with water, 5% aqueous NaHCO ₃ , water,
After washing with 10% aqueous citric acid and water successively, and drying, the solvent was distilled off 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 The chlorinated 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 distilled off under reduced pressure. Residue is silica gel (100 g)
The residue was purified by column chromatography (chloroform) using to give the azide compound 526 (2.98 g) 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), and a Lindlar catalyst (4.40 g) was added. Catalytic reduction was performed. After removing the catalyst by filtration, the filtrate was concentrated under reduced pressure to obtain an amine compound 527 (2.84 g) as a colorless oily substance.

4) Synthesis of Compound 528 The amine compound 527 (240 mg) and palmitic acid N-hydroxysuccinimide ester (143 mg) were dissolved in methylene chloride (20 ml), and triethylamine (82 mg) was added.
Stir at room temperature for 3 hours. The reaction solution was diluted with methylene chloride, washed sequentially with water, 10% aqueous citric acid and water, dried, and the solvent was distilled off under reduced pressure. The residue was purified by column chromatography (chloroform-methanol 100: 1) using silica gel (50 g) to obtain the desired 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 = 6.6 Hz), 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 stirred at room temperature for 2.5 hours. When the reaction mixture contains "Amberlite IRC
-50 "was added, the insolubles were removed by filtration, and the filtrate was concentrated to dryness under reduced pressure. Chloroform was added to the residue, the insoluble material was removed by filtration, and the filtrate was concentrated to dryness under reduced pressure. The residue was washed with ether to obtain the desired 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, m
d, J = 3.4Hz), 6.43 (1H, m).

Example 2 (Synthesis example (2)) 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.
243, 251, 830, 833, 856, 7
18, 1105, 1205, 4-3, 8-2, 12-8, 15-2, 2
8-2 and 18-2 were synthesized as follows.

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

(E) Synthesis of Compound 243 (FIG. 9) 1) Synthesis of Compound 240 Org, Chem. , 56 , 4326 (1991), 1,980 g of 2- {2- [2- (2-azidoethoxy) ethoxy] ethoxy} ethanol and 3.525 g of β-D-galacto-spentaacetate in 50 ml of methylene chloride.
And stirred under ice-cooling. To this, 4.44 ml of boron trifluoride diethyl ether complex was added dropwise. 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 a saturated saline solution, dried over magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue is purified by silica gel column chromatography (elution solvent: n-hexane-ethyl acetate 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 (c = 1.02, CHCl ₃ ).

2) Synthesis of compound 241 120 ml of ethyl acetate was added to 1.129 g of compound 240 and dissolved. Here, p-toluenesulfonic acid monohydrate 0.391 g
And 0.570 g of Lindlar's catalyst were added, and the catalyst was reduced at 50 psi for 5.5 hours. Further, 0.564 g of Lindlar's catalyst was added, and the catalyst was reduced at 50 psi for 5 hours. The catalyst was removed by filtration to give 1.172 g of the desired product as a pale brown oil. It was used for the following reaction without further purification.

3) Synthesis of Compound 1103 To 0.525 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 and filtered off under reduced pressure (three times). It was used for 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, followed by stirring under ice-cooling. Here 268μl of triethylamine
Was added, and the whole amount of the compound 1103 obtained in the above reaction was dissolved in 4 ml of methylene chloride, and the mixture was stirred for 14 hours while warming to room temperature. The mixture was diluted with methylene chloride, washed with water and saturated saline, 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 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 to 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 = 1.0 Hz, 3.5 Hz), 5.96 (t, 1H, J = 5.6 Hz).

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

5) Synthesis of Compound 243 Compound (242) (0.518 g) was added to methanol (5 ml) and benzene (10).
ml was added to dissolve. To this was added 6 drops of a 28% sodium methoxide methanol solution to adjust the pH to 10, followed by stirring at room temperature for 19 hours. A "Dowex 50X-8" ion exchange resin (H type) was added to neutralize the mixture, and the resin was removed by filtration. The solvent was distilled off under reduced pressure, and the residue was purified with "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 1.55 g (60% dispersion) of sodium hydride was washed with n-hexane and suspended in 40 ml of N, N-dimethylformamide. The mixture was stirred under ice cooling. To this, 4.47 ml of malonic acid dibenzyl ester was dissolved in 10 ml of N, N-dimethylformamide and added dropwise, followed by stirring at room temperature for 30 minutes. After cooling again with ice, 7.29 ml of n-octyl bromide was added, and the mixture was stirred at 60 ° C for 18 hours.
The solvent was removed by filtration under reduced pressure. Ethyl acetate and water were added to the residue,
The organic layer was separated. The extract was washed with saturated saline, dried over magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue is purified by silica gel column chromatography (elution solvent: n-
Hexane-ethyl acetate 20: 1) to obtain 7.227 g of the desired product as a colorless oil.

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 to 7.32 (m, 10
H).

2) Synthesis of Compound JJJ 70 ml of ethyl acetate was added to 6.967 g of the compound HHH to dissolve it, and 0.105 g (dry matter) of 10% Pd-C was added thereto, followed by catalytic reduction under normal pressure for 11.5 hours. The catalyst was removed by filtration, 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 JJJ : 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 was added 1.5 ml of thionyl chloride, and
Heated at C for 2.5 hours. Thionyl chloride was distilled off under reduced pressure. Benzene was added to the residue to dissolve it and distilled off under reduced pressure (three times). It was used for 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, followed by stirring under ice-cooling. 259μl of triethylamine
Was added, and the whole amount of the compound KKK obtained in the above reaction was dissolved in 5 ml of methylene chloride, and the mixture was stirred for 12 hours while warming to room temperature. The mixture was diluted with chloroform, washed with water and saturated saline, dried over magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel chromatography (elution solvent: n-hexane-ethyl acetate 1: 2) to obtain 0.357 g of the desired product as a colorless amorphous.

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, 10.7 Hz), 4.13 (dd, 1H, J = 7.0 Hz, 11.3 Hz), 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 To 0.325 g of Compound 250, 10 ml of methanol and benzene 5
ml was added to dissolve. To this, 4 drops of a 28% sodium methoxide methanol solution was added to adjust the pH to 11, and the mixture was stirred at room temperature for 14 hours. A "Dowex 50X-8" ion exchange resin (H type) was added to neutralize the mixture, and the resin was removed by filtration. The solvent was distilled off under reduced pressure, and the residue was purified with "Sephadex LH-20" (elution solvent: chloroform-methanol 1:
1), 0.235 g of the target compound was obtained as a colorless amorphous.

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 = 3.4 Hz), 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 to react. I let it.

The reaction solution was subjected to centrifugation, and methanol was added to the obtained insoluble material, followed by centrifugation again.
49 mg (yield 67%) of c derivative compound 830 was obtained.

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 to 4.50 (m, 3H).

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

(H) Synthesis of compound 833 (FIG. 12 ) To a solution of amide compound 832 (69 mg) in methanol (3 ml) was added sodium methoxide (6 μl, 28% methanol solution), and the mixture was stirred at room temperature for 3 hours to react. I let it.

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

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 filtered off under reduced pressure. (Twice). It was used for 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. 139μl of triethylamine here
Was added, and the whole amount of the compound obtained in the above reaction was dissolved in 5 ml of methylene chloride, and the mixture was stirred for 3 days while warming to room temperature. The mixture was diluted with methylene chloride, washed with water and saturated saline, dried over magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue is purified by silica gel column chromatography (elution solvent: methylene chloride-methanol 70:
1) 0.318 g of the target product was obtained as a colorless amorphous.

By ¹ H-NMR, a mixture of about 1: 1 of two kinds of conformers was obtained.
One hydrogen of the conformer is shown counted 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 was mixed with 0.300 g of benzene 4 ml and methanol 2
The mixture was added and dissolved, and stirred under ice cooling. To this was added 3 drops of a 28% sodium methoxide methanol solution to adjust the 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 removed by filtration. The solvent was distilled off under reduced pressure, and the residue was purified by "Sephadex LH-20" (elution solvent: chloroform-methanol 1: 1), and 0.147 g of the target compound was obtained as a colorless powder.
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-lactosoctoacetate (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 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), it was washed with saturated saline, dried over magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue is purified by silica gel column chromatography (elution solvent: n-hexane-ethyl acetate 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 716 150 ml of ethyl acetate was added to 2.69 g of compound 715 and dissolved. 0.65 g of p-toluenesulfonic acid monohydrate and 1.32 g of Lindlar catalyst were added thereto, and the mixture was catalytically reduced at 50 psi for 4.5 hours. Add 1.31g of Lindlar catalyst and add 50ps
i for 2.5 hours. The catalyst was removed by filtration to obtain 3.11 g of the desired product as a pale brown oil. The following reaction was used without further purification.

4) Synthesis of Compound 717 To 1.51 g of Compound 716, 20 mL of methylene chloride and 220 μL of triethylamine were added and dissolved. The entire amount of Compound 1103 obtained by the above reaction was dissolved in 5 mL of methylene chloride, and the mixture was further stirred for 19.5 hours. did. The mixture was diluted with methylene chloride, washed with 10% citric acid, water and saturated saline, dried over magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (eluent: n-hexane-ethyl acetate 1: 2) to give 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 = 5.5 Hz).

[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. Five drops of a 28% sodium methoxide methanol solution were added thereto 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 removed by filtration. The solvent was distilled off under reduced pressure, and the residue was purified by "Sephadex LH-20" (elution 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 = 7.5 Hz), 5.06 (d, 1H, J = 8.0 Hz), 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 β-D-glucose pentaacetate 6.16 g 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 methene 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 with water four times (the aqueous layer became neutral), wash with saturated saline,
After drying over magnesium sulfate, the solvent was distilled off under reduced pressure.
The residue was purified by silica gel column chromatography (elution 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 compound 1101, 150 ml of ethyl acetate was added and dissolved. 0.79 g of p-toluenesulfonic acid monohydrate and 1.04 g of Lindlar catalyst were added thereto, and the mixture was subjected to catalytic reduction at 50 psi for 6.5 hours. Add 1.04g of Lindlar catalyst and add 50ps
Catalytic reduction with i for 3 hours. The catalyst was removed by filtration to give 2.49 g of the desired product as a pale brown oil. Without further purification
The following reaction was used.

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 refluxed for 6 hours. Thionyl chloride was distilled off under reduced pressure. Benzene was added to the residue to dissolve it, and evaporated under reduced pressure (twice). It was used for 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. The entire amount of compound 1103 obtained in the above reaction was dissolved in 5 ml of methylene chloride, and the mixture was further stirred for 2 hours. And further stirred at room temperature for 30 minutes. The mixture was diluted with methylene chloride, washed with 1N hydrochloric acid, water and saturated saline, 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),
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, 1H
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 To 0.62 g of compound 1104, 6 ml of methanol and 12 ml of benzene were added.
Was added and dissolved, and the mixture was stirred under ice cooling. Five drops of a 28% sodium methoxide methanol solution were added thereto 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 removed by filtration. The solvent was distilled off under reduced pressure, and the residue was purified by "Sephadex LH-20" (elution 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-Ribosetetraacetate (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 stirred under ice cooling. Here is the 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 washing with water (the aqueous layer became neutral), the mixture was washed with saturated saline, 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),
1.967 g of the desired product was obtained as a colorless oil.

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 1.844 g of Compound 1202 was dissolved by adding 100 ml of ethyl acetate. Here, 0.809 g of p-toluenesulfonic acid monohydrate
And 0.944 g of Lindlar's catalyst, and the mixture was catalytically reduced at 50 psi for 4 hours. Further, 0.855 g of Lindlar's catalyst was added,
Catalytic reduction was performed at 50 psi for 3 hours. The catalyst was removed by filtration to obtain 2.422 g of the desired product as a pale brown oil. It was used for 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 refluxed for 2.5 hours. Thionyl chloride was distilled off under reduced pressure. Benzene was added to the residue to dissolve it and distilled off under reduced pressure (three times). It was used for the following reaction without further purification.

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

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 0.477 g of compound 1204, 3 ml of methanol and 6 parts of benzene were added.
ml was added to dissolve. 6 drops of a 28% sodium methoxide methanol solution are added thereto to adjust the pH to 12, and the
Stirred for hours. A "Dowex 50X-8" ion exchange resin (H type) was added to neutralize the mixture, and the resin was removed by filtration. The solvent was distilled off under reduced pressure, and the residue was separated using "Sephadex LH-20".
(Elution solvent: chloroform-methanol 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, 1H
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 an alcohol (compound 3-
1) (8.04 g) was dissolved in methylene chloride (180 ml),
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%
After washing with NaHCO ₃ water and water sequentially and drying, the solvent was distilled off under reduced pressure. The residue was purified twice by column chromatography (chloroform) using silica gel (200 g).
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 = 10.02H)
z), 4.88 (1H, dd, J = 3.67Hz, 10.26H
z), 5.17 (1H, dd, J = 9.77 Hz, 10.02H
z), 5.21 (1H, d, J = 3.67 Hz), 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 and dried, and the solvent was distilled off under reduced pressure. The residue was purified by column chromatography (chloroform) using silica gel (200 g) to give the desired compound 3-4 (2.10 g) as a colorless oil.

¹ H-NMR (CDCl ₃ ) δ: 2.02 (3
H, s), 2.03 (3H, s), 2.06 (3H, s), 3.40
(2H, t, J = 5.13 Hz), 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 =
9.77 Hz, 10.26 Hz), 5.21 (1H, d, J = 3.66H)
z), 5.54 (1H, dd, J = 9.77 Hz, 10.26 H
z). (iii) Synthesis of Compound 3-5 Compound 3-4 (1.70 g) and p-toluenesulfonic acid monohydrate (0.66 g) were dissolved in ethanol (100 ml), and a Lindlar catalyst (3.00 g) was added. Catalytic reduction was performed at 40 psi for 5 hours.

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

(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)
Of methylene chloride (20 ml) was stirred at room temperature for 20 hours.
To the reaction solution, an amine compound (compound 3-5) (364 mg) dissolved in methylene chloride (5 ml) was added, and then triethylamine (115 mg) was added, followed by stirring at room temperature for 3 hours.

The reaction solution was diluted with chloroform and diluted with water, 5%
After washing with NaHCO ₃ aqueous solution, water, citric acid aqueous solution and water successively and drying, the solvent was distilled off under reduced pressure. The residue was purified by column chromatography (hexane-ethyl acetate 1: 1) using silica gel (30 g) to obtain the desired 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 =
6.59 Hz), 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 = 3.66 Hz, 10.26 Hz), 5.17 (1
H, dd, J = 9.28 Hz, 10.26 Hz), 5.24 (1H,
d, J = 3.66 Hz), 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 stirred at room temperature for 24 hours. The reaction solution was concentrated under reduced pressure, and methanol (3 ml) and 0.1 N aqueous NaOH (2 ml) were added to the residue.
Stirred at room temperature for 6 hours.

The reaction mixture 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 (6H, t, J =
6.84 Hz), 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 between 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 and diluted with water, 5%
After washing with NaHCO ₃ water and water sequentially and drying, the solvent was distilled off under reduced pressure. The residue was subjected to column chromatography using silica gel (150 g) (chloroform-methanol 200: 1).
→ Purified 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 (1H, dd, J
= 11.70 Hz, 12.95 Hz), 1.89 (3H, s), 2.01
(3H, s), 2.03 (3H, s), 2.06 (3H, s),
2.15 (3H, s), 2.43 (1H, dd, J = 4.88 Hz,
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 = 2.44 Hz, 12.4
6 Hz), 5.25 (1H, ddd, J = 4.88 Hz, 10.99
Hz, 11.70 Hz), 5.28 (1H, ddd, J = 2.44H)
z, 3.42 Hz, 8.30 Hz), 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 was silica gel (50
g) and purified by column chromatography (chloroform-methanol 150: 1) using the desired compound 7-3.
(510 mg) was obtained as a colorless oil.

[Α] _D + 2.6 ° (c 1.02, CHCl ₃ ). ¹ H-NMR (CDCl ₃ ) δ: 1.89 (1H, dd, J
= 11.70 Hz, 12.94 Hz), 1.89 (3H, s), 2.01
(3H, s), 2.03 (3H, s), 2.06 (3H, s),
2.15 (3H, s), 2.44 (1H, dd, J = 4.89 Hz,
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 = 8.55 Hz, 12.46 Hz), 4.14 (1H, ddd, J
= 10.02 Hz, 10.50 Hz, 10.75 Hz), 4.46 (1
H, dd, J = 2.20 Hz, 10.50 Hz), 4.90 (1H,
dd, J = 2.44 Hz, 12.46 Hz), 5.25 (1H, dd)
d, J = 4.89 Hz, 10.75 Hz, 11.70 Hz), 5.27
(1H, ddd, J = 2.44Hz, 3.42Hz, 8.55H
z), 5.40 (1H, dd, J = 2.20 Hz, 3.42 Hz),
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), and a Lindlar catalyst (1.50 g) was added. The catalytic reduction was performed for 7 hours. After removing the catalyst by filtration, the filtrate was concentrated under reduced pressure to obtain the desired compound (590 mg).

(Iv) Synthesis of compound 8-1 Methylene chloride (40 ml) of carboxylic acid (compound 4-1) (153 mg), N-hydroxysuccinimide (35 mg) and N, N'-dicyclohexylcarbodiimide (62 mg) And a mixed solution of 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, and then triethylamine (53 mg) was added and the mixture was stirred at room temperature for 24 hours. The reaction solution was concentrated under reduced pressure, diluted with chloroform, and water, 5% NaHCO ₃
The extract was washed sequentially with water, water, citric acid water and water, dried, and the solvent was distilled off under reduced pressure. The residue was subjected to column chromatography using silica gel (50 g) (chloroform-methanol).
Purification by 150: 1) gave the desired 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 =
6.84 Hz), 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 (1H, dd, J = 11.48 Hz, 12.94H
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 = 8.55 Hz, 12.46 Hz), 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 = 2.44 Hz, 12.46 H)
z), 5.25 (1H, ddd, J = 2.44 Hz, 3.42 Hz,
8.55 Hz), 5.28 (1H, ddd, J = 4.89 Hz, 10.7
5 Hz, 11.48 Hz), 5.41 (1H, dd, J = 2.44H)
z, 3.42 Hz), 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% NaOMein in MeOH (20 µl) was added, and the mixture was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, the residue was dissolved in methanol (12 ml), 0.1 N aqueous NaOH (3 ml) was added, and the mixture was stirred at room temperature for 16 hours. After heating the reaction solution, "Amberlite IRC-50" was added, the insoluble material was removed by filtration, 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 = 6.59 Hz), 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) A mixture of 50 g (86.7 mmol), 2 ml of 28% sodium methylate and 600 ml of methanol was stirred at room temperature for 4 hours and treated in a conventional manner to give 1,6-anhydrolactose (compound 12-2) ( 28.5 g).

[Α] _D -46.5 ° (c 1.0, H ₂ O). Compound 12-2 (16.2 g) was dissolved in 150 ml of pyridine, and trityl chloride (20.9 g) was added, followed by a reaction at 50 ° C. for 2 hours. Then, a pyridine solution of benzoyl chloride (5
0.6 g) and 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 obtain compound 12-3.
(35 g) was obtained.

The elementary analysis value of C ₆₆ H ₅₄ O ₁₅ was calculated: C, 72.93; H, 5.01, actually measured value, C, 72.63;
H, 5.30.

[Α] _D + 54.2 ° (c 1.0, CHCl ₃ ). _{^{1 H-NMR (CDCl 3)}} : 3.16 (1H, 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 solution was neutralized with saturated aqueous sodium hydrogen carbonate, the solvent was distilled off, extracted with chloroform, and purified by silica gel column chromatography (ethyl acetate-hexane, 1: 1) to obtain compound 12-4 (18.6 g). ) Got.

Elemental analysis value of C ₄₇ H ₄₀ O ₁₅ was calculated: C, 66.86; H, 4.77, actually measured: 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, cooled with ice, added with diphenyl phosphorochloridate (8.27 g), returned to room temperature, and stirred for 1 hour.

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

The elemental analysis value of C ₅₉ H ₄₉ O ₁₈ P was calculated: C, 65.79; H, 4.59; P, 2.88, but found 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 = 7.6 Hz), 4.04 (1H, ddd, J
= 10.5Hz, 7.5Hz, 7.0Hz), 4.22 (1H, dd
d, J = 10.5 Hz, 8.0 Hz, 7.0 Hz), 4.34 (1
H, t, J = 6.0 Hz), 4.56 (1 H, d, J = 5.4 H)
z), 4.96 (1H, s), 5.31 (1H, d, J = 8.1H)
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 (35H, m). (iv) Synthesis of compound 12-6 A mixture of compound 12-5 (18.2 g), trifluoroacetic acid 29 ml and acetic anhydride 356 ml was stirred at room temperature for 20 hours.

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

The elemental analysis value of C ₆₃ H ₅₅ O ₂₁ P was calculated as follows: C, 64.18; H, 4.70; P, 2.63;
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 = 10.5 Hz, 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 = 10.3 Hz), 4.22 (dd, J =
11.4 Hz, 3.4 Hz), 4.27 (dd, J = 11.4 Hz, 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 = 10.5 Hz, 7.0 Hz), 3.56
(Dt, J = 10.5Hz, 10.5Hz, 7.0Hz), 3.77
(M), 4.71 (d, J = 7.8 Hz), 5.50 (dd, J =
10.0 Hz, 8.3 Hz), 5.64 (dd, J = 10.5 Hz, 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), 150 ml of methylene chloride and "Molecular Sieve 4A" (MS4A) was ice-cooled, and trimethylsilyl triflic was obtained. A 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 bicarbonate, 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. Elementary analysis as C ₈₅ H ₁₀₁ O ₂₃ P is Calculated: C,
67.08; H, 6.69 found: 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
= 10.0 Hz, 3.0 Hz), 5.37 (1H, dd, J = 10.0)
Hz, 7.9 Hz), 5.64 (1H, dd, J = 10.0 Hz,
7.9 Hz), 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 normal pressure for 20 hours using platinum oxide as a catalyst.
The catalyst was filtered off, neutralized with 28% sodium methylate (p
H6) The solvent was distilled off. The residual solid was dissolved in benzene-methanol (1: 1, 24 ml) and 28% sodium methylate was dissolved.
0.6 ml was added and the mixture was stirred at room temperature for 20 hours.

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

Mp: 195-199 ° C (dec). C ₃₆ H ₆₉ O ₁₇ PNa Elementary analysis as ₂ · 4H ₂ O is Calculated: C, 46.85; H, 8.40 ; P, 3.36; Na,
Found 4.98; C, 47.06; H, 8.32; P, 3.31; N
a, 4.83.

[Α] _D −4.5 ° (c 0.8, H ₂ O). ¹ H-NMR (D ₂ O): 0.91 (3H, 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) ) In methylene chloride (300 ml) was added with a solution of a 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 extracted by adding 200 ml of chloroform. The organic layer was washed four times with water and dried over anhydrous sodium sulfate. The solvent was concentrated under reduced pressure, and the residue was separated by 1700 ml of 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
0.2153 (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 and stirred at 60 ° C. for 17 hours.

To the obtained solution was added 100 ml of water, and the mixture was extracted with ethyl acetate. The organic layer was washed three times with water and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was separated by 350 ml of 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 treated with a sodium methylate methanol solution (5 mol / l).
Was added and stirred at room temperature for 3 hours. The resulting solution was neutralized by adding an acidic ion exchange resin “DOWEX 50W × 8”, and the resin was separated by filtration, 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 and stirred 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 to dissolve the residue.
The mixture was heated and stirred at 50 ° C. for 5 hours.

The obtained solution was neutralized with triethylamine, and the solvent was distilled off 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 removed.
Separation by 500 ml of silica gel column chromatography (hexane: ethyl acetate = 2: 1-1: 1) gave the desired product. 2.4106 g, 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 chloride (1.493 g) in pyridine (3 ml) under ice cooling, and the mixture was stirred at room temperature overnight. . The obtained 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 saline, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was separated by 500 ml of 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.
g) was added and the mixture was stirred for 6 hours under a 50 psi hydrogen atmosphere. Further, a Lindlar catalyst (1.0 g) was added, and the mixture was stirred under a 50 psi hydrogen atmosphere 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-palmityl stearic acid (1.354 g), N-hydroxysuccinimide (306 mg)
And a mixture of triethylamine (0.683 ml) were dissolved in 40 ml of methylene chloride and 20 ml of hexane.
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 silica gel column chromatography of 200 ml (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 50 mg of platinum oxide were added 20 ml of ethyl acetate and 10 ml of methanol, 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. 2 ml of benzene and 6 ml of methanol were added to the residue and dissolved, and a methanol solution of sodium methylate (5 mol / mol) was added.
1) The pH was adjusted to 11 by adding 30 drops, and the mixture was stirred at room temperature overnight.

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 column chromatography on 150 ml of silica gel (chloroform: methanol: water = gradient elution from 65: 25: 3 to 60: 35: 7, (Total 2000 ml, 100 fractions). 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 the resin was filtered off. The solvent is concentrated under reduced pressure,
It was dissolved in a mixed solvent of chloroform: methanol = 9: 1, and the mixture was dissolved in “Sephadex LH-20” (22 mmφ × 400 mm).
, Chloroform: methanol = 9: 1). The desired product was obtained by concentrating the fraction 5-10.
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.
Reflux under heating for hours. Thionyl chloride is distilled off under reduced pressure,
Benzene was added to the residue and evaporated under reduced pressure (twice). It
It was used for 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, followed by stirring under ice-cooling. Here triethylamine 350 μ
and the whole amount of the compound 11-4 obtained in (a) is dissolved in 5 ml of methylene chloride and added.
Stirred for hours.

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), 1.23
2 g were 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,
11.5 Hz, 8.7 Hz), 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, 30 ml of tetrahydrofuran and 10 ml of methanol were added and dissolved. Here platinum oxide 0.01
5 g was added, and the mixture was catalytically reduced at normal pressure for 6 days. The catalyst was removed by filtration, and the solvent was distilled off under reduced pressure. 8 ml of benzene and 8 ml of methanol were added to the residue to dissolve it, and 22 drops of a 28% sodium methoxide methanol solution were added (pH = 11), followed by stirring at room temperature for 11.5 hours.

After cooling with ice and neutralizing with 1N hydrochloric acid, the solvent was distilled off under reduced pressure. The residue was purified on a "Sephadex LH-20" column (resin: about 150 ml, elution solvent: chloroform-methanol-water 10: 10: 3). 0.640 g of the desired 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 and 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 extracted with chloroform (150 ml). 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 of 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 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 of silica gel column chromatography (hexane: ethyl acetate = 2: 1.5-1: 1) to obtain the desired product. 5.30 g.

[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
1.2 Hz), 4.176 (dd, 1H, J = 6.3 Hz), 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 a sodium methylate methanol solution (5 mol / l).
10 drops were added and 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 distilled off under reduced pressure. The residue was dissolved in pyridine (30 ml), and to this solution was added diphenyl phosphate 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 stirred overnight.

The solution was added to ice water, and chloroform (200 m
l) Extracted in The organic layer was washed twice with 2N hydrochloric acid and three times with saturated saline, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was separated by 500 ml of silica gel column chromatography (hexane: ethyl acetate =
2: 1-1: 1) to obtain the desired product. 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 = 7.1H)
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 = 10.5 Hz), 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), and Lindlar catalyst ( 1.2 g) was added, and the mixture was stirred under a 50 psi hydrogen atmosphere for 7 hours.

The catalyst was filtered off, 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-palmityl stearic acid (1.188 g), N-hydroxysuccinimide (269 mg)
And 4-dimethylaminopyridine (523 mg) in DMF
(100 ml), N, N'-dicyclohexylcarbodiimide (482 mg) was added to the 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 deposited N, N'-dicyclohexylurea was filtered off and the solvent was distilled off at 40.degree. Ethyl acetate (40 ml) was added to the residue, and insolubles were removed by filtration. The filtrate was concentrated under reduced pressure, and the residue was separated by 200 ml of 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,
10.7 Hz, 8.5 Hz), 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 = 10.4 Hz), 5.778 (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 normal pressure hydrogen atmosphere. 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 a sodium methylate methanol solution (5 mol / l) were added to adjust the pH to 11, followed by stirring at room temperature for 5 hours.

After neutralizing the solution with 1N hydrochloric acid, the solvent was distilled off under reduced pressure. Residue in chloroform: methanol =
It was dissolved in a 1: 1 mixed solvent, 2.5 g of silica gel was added, and the solvent was distilled off under reduced pressure. The dried silica gel obtained
150 ml of chloroform: methanol: water = 65: 25: 4
And a fraction containing the target substance was collected (22 mmφ × 400 mm, chloroform: methanol: water = 65: 25: 4 to 2: 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 “Sephadex LH-20” was packed with the solvent.
(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) Among the compounds of the present invention synthesized in Examples 1 and 2,
Liposomes were prepared as follows using each of a total of 16 compounds except 1205 and 28-2.

80 μmol of L-α-dipalmitoylphosphatidylcholine, 80 μmol of cholesterol, 8 μmol of dicetyl phosphate and 16 μmol of the compound of the present invention were dissolved in a chloroform / methanol filtrate (1: 1 by volume).
Next, the organic solvent was removed in a stream of nitrogen gas 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 slightly sonicated 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 be converted into liposomes in the same manner.

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

Example 4 (Evaluation Test) Samples The four compounds of the invention and the four control compounds synthesized in Example 1 were used instead of 1 mM inulin for ³
A total of eight kinds 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 Eight kinds of prepared samples were each used for SD male rats (body weight).
200-250 g) L-α per 100 g body weight from hind limb vein
5 μmol of dipalmitoylphosphatidylcholine and cholesterol were injected in total.

Six hours after the administration, the rats were sacrificed, about 200 mg of various tissues were taken, dried and burned by a combustion device, and the radioactivity was determined by a liquid scintillation method.
The inulin concentration per g was determined.

C. Results The results are shown in FIGS.

From these results, it can be seen that the liposome containing the lipid derivative having at least two alkyl groups (the present invention)
Shows improved organ recognition compared to liposomes (control) produced by the same method as the method of the present invention except that a compound having one alkyl group was used.

[0278]

According to the present invention, a liposome having excellent stability, excellent organ directivity, and excellent drug-retaining function can be easily provided.

[Brief description of the drawings]

FIG. 1 shows the reaction in Example 1.

FIG. 2 shows a reaction in Example 1.

FIG. 3 shows a reaction in Example 1.

FIG. 4 shows a reaction in Example 1.

FIG. 5 shows a reaction in Example 1.

FIG. 6 shows a reaction in Example 1.

FIG. 7 shows a reaction in Example 1.

FIG. 8 shows the reaction in Example 1.

FIG. 9 shows a reaction in Example 2.

FIG. 10 shows the reaction in Example 2.

FIG. 11 shows a reaction in Example 2.

FIG. 12 shows a reaction in Example 2.

FIG. 13 shows a reaction in Example 2.

FIG. 14 shows a reaction in Example 2.

FIG. 15 shows a reaction in Example 2.

FIG. 16 shows a reaction in Example 2.

FIG. 17 shows a reaction in Example 2.

FIG. 18 shows a reaction in Example 2.

FIG. 19 shows a reaction in Example 2.

FIG. 20 shows a reaction in Example 2.

FIG. 21 shows a reaction in Example 2.

FIG. 22 shows a reaction in Example 2.

FIG. 23 shows the results in Example 4.

FIG. 24 shows the results in Example 4.

FIG. 25 shows the results of Example 4.

FIG. 26 shows the results of Example 4.

──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Shiro Miyoshi 206-1 Aku, Mishima-shi, Shizuoka Prefecture No.5 Tamura Heights 5 (72) Inventor Katsutoshi Aono 2-529-4 Gakuen Asahimotomachi, Nara-shi, Nara Excel Heights B-308 (72) Inventor Hitoshi Yamauchi 2-32-12 Okudo, Katsushika-ku, Tokyo (72) Inventor Naoichi Murahashi 7-2-4, Matsumaedai, Moriyacho, Kitasoma-gun, Ibaraki Prefecture (72) Inventor Atsushi Sasaki, Tsukuba City, Ibaraki Prefecture 4-19-13 Kasuga Eisa I Shizan Ryo 307 (72) Inventor Hiroshi Watanabe 7-131 Seven Pier 301, Shinmatsudo, Matsudo-shi, Chiba (72) Inventor Hideo Kaneko 1-1-25 Nakamuracho, Minami-ku, Yokohama-shi, Kanagawa-ken ( 58) Surveyed fields (Int.Cl. ⁶ , DB name) A61K 9/127 A61K 47/28

Claims (4)

(57) [Claims] As raw materials, at least (1) 1 mol of a polar lipid, (2) a compound imparting a positive or negative charge of 0.05
~ 0.5 mol, (3) cholesterol 0.3-1.5 mol, (4)
Organ directional sensor in the molecule, compounds 0.02 to 0.5 mole with a polyethylene glycol and at least two alkyl groups of carbon number 5-20 of polymerization degree 3-6 and (5) the aqueous solvent 50 to 100 l at this ratio A method for producing a liposome, which is used. Wherein (1) polar lipids 1 mol, (2) Compound 0.05 mole which gives a positive or negative charge, (3) cholesterol 0.3-1.5 mol, and (4) Organ directional sensor in the molecule
Compounds having a polyethylene glycol and at least two alkyl groups having a carbon number of 5-20 Sir polymerization degree 3-6 0.
The solvent is removed from an organic solvent containing 02 to 0.5 mol in this ratio to form a lipid film, and then an aqueous solution of the drug to be included in the liposome is added to the lipid film in an amount of 50 to 100 l per mol of polar lipid. 2. The method for producing liposomes according to claim 1, further comprising forming liposomes and selectively collecting liposomes having a particle size of 50 to 1000 nm. (3 ) 1 mol of a polar lipid, (2) 0.05 to 0.5 mol of a compound imparting a positive or negative charge, (3) 0.3 to 1.5 mol of cholesterol , and (4) an organ-directing sensor in a molecule.
Sir, compounds having a polyethylene glycol and at least two alkyl groups of carbon number 5-20 of polymerization degree 3-6
50 to 100 l of an aqueous solution of the drug to be included in the liposome is added to an organic solvent containing 0.02 to 0.5 mol in this ratio to 1 mol of the polar lipid, and then the mixed solution is subjected to ultrasonic treatment to obtain a w / o-type mixture. 2. The liposome production according to claim 1, wherein the organic solvent is removed under reduced pressure after forming the emulsion, and the organic solvent is removed again under reduced pressure as needed after performing vortexing to form an o / w emulsion. Law. (4) As a raw material, at least (1) 1 mol of a polar lipid, (2) a compound which gives a positive charge or a negative charge.
05 to 0.5 mol, (3) cholesterol from 0.3 to 1.5 mol list
To (4) Organ directional sensor in the molecule, polyethylene glycol having a degree of polymerization of 3-6 and at least two carbon number 5
A liposome containing 0.02 to 0.5 mol of a compound having 20 alkyl groups in this ratio, and using an organic solvent containing a drug and 50 to 100 l of an aqueous solvent per 1 mol of polar lipid in this ratio. Manufacturing method.