JPS6351396A - Lewis type-b glycolipid and production thereof - Google Patents

Abstract

NEW MATERIAL:The compound of formula I [R<1> is H or acetyl; R<2> is OH, acetyloxy, F or group of formula II (R<3> is H or benzoyl); Ac is acetyl]. USE:Agent for diagnosis, examination and remedy for various cancers. PREPARATION:The compound of formula III (Bn is benzyl) is reduced with hydrogen in the presence of a catalyst (preferably Pd-C, Pd-BaSO4, PtO2 or Pt-C).

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a novel glycolipid having an epitope and a method for producing the same.

[Conventional technology]

Glycolipids are a group of constituent components of the lipid bilayer that forms cell membranes.In the cell membrane, the lipid portion is embedded in the double membrane layer, and the sugar chain portion is directed toward the outside of the cell. The types of these glycolipids and the content of each glycolipid in the cell membrane are determined by cell type, tissue,
Different for each animal species.

It is also known that the carbohydrate pattern differs depending on the stage of development, differentiation, etc.

On the other hand, recently established techniques for producing monoclonal antibodies are
It is increasingly being applied to antibody production in fields such as immunology, embryology, and oncology. Among these, the antigenic substances of antibodies that exhibit cancer specificity are often sugar chains when searched at the human science level.

It is known to be one of the antigenic determinants (epitopes) of various blood group substances, which are a group of 7-1-substituted lacto-sphingoglycosphingolipids. Furthermore, many glycolipids having such epitopes are found in abnormal glycolipids that occur in cancerous cells such as gastric cancer, colon cancer, and pancreatic cancer. Therefore, research on the isolation and identification of glycolipids having epitopes has recently been actively conducted.

However, these glycolipids that can be isolated from nature are only in trace amounts and often contain impurities.

Based on these facts, it is of various significance to provide a method for obtaining organically pure glycosphingolipids of the fuyuse-substituted lactone series. That is, it can provide a means to elucidate the biological functions of cancer-related glycolipids due to cancerous changes. Furthermore, glycolipids can be used to diagnose various cancers,
It can be applied to inspection and treatment.

In particular, Lewis b antigen is known as a cell surface antigen of human colon cancer [J
, Biol, Chem, 256 (1981) 1
3223-13225).

Therefore, by establishing a synthetic route for the antigen, it is possible to synthesize the antigen more easily than by isolating it from natural products.
It is expected that the synthetic antigen can be applied to diagnosis and testing techniques for diagnosing malignant tumors.

[Purpose of the invention]

An object of the present invention is to provide a novel glycolipid having an epitope and a method for producing the same.

[Structure of the invention]

That is, the present invention relates to a compound represented by general formula [I].

(In the formula, R' is a hydrogen atom or an acetyl group, and R2
is a hydroxyl group, an acetoxy group, a fluorine atom, an -R3 atom, or a benzoyl group), and AC represents an acetyl group. ) Specific examples of the present invention are shown below.

R'[] Ac Niacetyl group, Bz: Benzoyl group The method for producing the compound of the present invention will be explained below.

First, a method for synthesizing starting compound (11), which is a hexasaccharide, using compound (1) as a starting material will be explained according to Schemes 1 and 2.

[Synthesis of trisaccharide compound (4)] Trisaccharide compound (4) is synthesized from trisaccharide compound (1) to compound (2) using D-lactose as a raw material by known means.
obtained via.

■Synthesis of compound (1) Compound (1) was prepared by Ogawa et al.
moto M,) is Carbohydrate Research (Ca
rbohydrateReserc, h) 135 (
It can be easily obtained from D-lactose by the method described in C3-C9 (1985).

■Synthesis of compound (2) Compound (2) can be obtained by reacting compound (1) with benzaldehyde dimethyl acecool in the presence of an acid catalyst, and then reacting the reaction product with a reducing agent in the presence of an acid catalyst. Can be done. The reaction with benzaldehyde dimethyl acecool is THF, dioxane, toluene, and DMF.
for 1 hour to 2 hours at a temperature of -10°C to 120°C in a solvent such as
It is preferable to carry out the reaction under stirring for several days. Examples of acid catalysts used in the reaction with benzaldehyde dimethyl acecool include p-)luenesulfonic acid, anhydrous hydrochloric acid, and benzenesulfonic acid. Furthermore, as a reducing agent, borane-trimethylamine complex (BH3・Me3
N), NaBCNH3, L+Af84, etc. Examples of the acid catalyst used in the reaction with the reducing agent include aluminum chloride, zinc chloride, tin chloride, anhydrous hydrochloric acid, and the like.

Incidentally, after the reaction with benzaldehyde dimethyl acecool is completed, the reaction product can be appropriately purified by known means such as column chromatography, and then subjected to the reaction with a reducing agent. At that time, the reaction mixture can also be neutralized with triethylamine or the like before purification.

■Synthesis of compound (4) Compound (4) is obtained by reacting compound (2) with compound (3) in the presence of a glycosidation catalyst such as ', 4eOTf, etc., and then reacting the reaction product with alkali methoxide. can be obtained by Compounds (2) and (3)
The reaction is dichloroethane, dichloromethane, nitromethane, acetonitrile, ethenobetoluene, THF,
It is preferable to carry out the reaction in a solvent such as dioxane at a temperature of -10°C to 1(10)°C with stirring for 5 minutes to 1 day. Further, the reaction with alkali methoxide is preferably carried out in a solvent such as methanol or ethanol at a temperature of -10°C to 80°C with stirring for 5 minutes to 1 day.

Compound (3) can be synthesized according to Scheme 5 using compound (A) as a raw material as follows.

(2) Synthesis of Compound (B) Compound G) can be obtained by reacting compound <A) with methylthiodo IJ 71-butyltin in the presence of 5na4. The reaction is carried out at -10°C in a solvent such as dichloromethane, dichloroethane, THF, or chloroform.
It is preferably carried out by stirring at a temperature of ~80°C for 30 minutes to 1 day.

■Synthesis of compound (C) Compound (C) is obtained by reacting compound (J3) with an alkali methoxide such as Na○), 10, etc. in a solvent such as Mecnor/Beethanol, and then reacting the resulting reaction product with T )IF,
It can be obtained by reacting with benzaldehyde dimethyl acecool in the presence of an acid catalyst such as p-toluenesulfonic acid in a solvent such as dioxane, toluene, or DMF. In addition, the reaction with alkali methoxide is -10~
It is preferable to carry out the reaction by stirring at 80° C. for 5 minutes to 1 day. Further, the reaction with benzaldehyde dimethyl acecool is preferably carried out by stirring at -10°C to 120°C for 1 hour to 2 days.

■Synthesis of Compound (D) Compound (C) can be synthesized by reacting Compound (C) with a reducing agent such as borane-trimethylamine complex in the presence of an acid catalyst such as aluminum chloride. -1 in a solvent such as THF, dioxane, toluene, etc.
It is preferable to carry out the reaction by stirring at 0°C to 80°C for 1 hour to 2 days.

(2) Compound (3) Compound (3) can be obtained by reacting compound (D) with acetic anhydride in the presence of 4-114AP.

The reaction is preferably carried out in a solvent such as pyridine, chloroform, dichloromethane, dichloroethane, THF, etc. by stirring at -10°C to 80°C for 30 minutes to 2 days.

[Synthesis of Tetrasaccharide Compound (7)] Tetrasaccharide compound (7) can be synthesized via trisaccharide compound (5) as shown in Schemes 1 and 2.

■Synthesis of compound (5) Compound [5] can be obtained by reacting compound (4) with hydrazine in a solvent such as ethanol, and then reacting the reaction product with acetic anhydride in a solvent such as methanol. Obtainable.

The reaction with hydrazine is preferably carried out at 0°C to 120°C with stirring for 5 minutes to 2 days. The reaction with acetic anhydride is preferably carried out at -10°C to 80°C with stirring for 5 minutes to 1 day.

(4) Synthesis of Compound (7) Compound (7) can be obtained by reacting compound (5) with compound (6) in the presence of a glycosidation catalyst. Examples of glycosidation catalysts include H
Mercury-based catalysts such as gBr2, Hg (CN)2 and Ag0
Examples include silver thread catalysts such as Tf and Ag silicate. Further, the reaction is carried out in a solvent such as dichloroethane, dichloromethane, nitromethane, toluene, acetonitrinope, THF, etc. under an inert atmosphere such as argon gas, from -10°C to
It is preferable to carry out the reaction by stirring at 1 (10)° C. for 1 hour to 2 days.

In addition, compound (6) is compound (E) according to scheme 6.
It can be synthesized as follows using as a raw material.

■Synthesis of compound (6) Compound (6) is obtained by reacting compound <E) with a reducing agent such as borane-trimethylamine complex in the presence of an acid catalyst such as aluminum chloride, and then converting the reaction product into the presence of 4-DMAP or the like. It can be obtained by reacting with acetic anhydride. The reaction with the reducing agent is T) (F) in a solvent such as dioxane or toluene at a temperature of -10°C to 80°C for 30 minutes to 2
It is preferable to carry out the reaction under stirring for several days. In addition, the reaction with acetic anhydride is carried out in a solvent such as pyridine, chloroform, dichloromethane, dichloroethane, THF, etc. at -10°C to 80°C.
It is preferable to carry out the reaction by stirring for 0 minutes to 2 days.

[Synthesis of hexasaccharide compound (U)]

Hexasaccharide compound (11) is compound (7) according to Scheme 2
Synthesized using raw materials.

■Synthesis of Compound (8) Compound (8) is produced by reacting Compound (7) with an alkali methoxide such as sodium methoxide in a solvent such as methanol or ethanol, and then converting the reaction product to toluenesulfonic acid, benzenesulfonic acid, It can be obtained by reacting with dimethoxypropane in the presence of an acid catalyst such as anhydrous hydrochloric acid or zinc chloride. The reaction with alkali methoxide is -1
It is preferable to conduct the reaction with stirring at a temperature of 0° C. to 80° C. for 5 minutes to 1 day.

Furthermore, the reaction with dimethoxypropane is acetone, TH
F, DMF,) in a solvent such as toluene, dioxane, etc., from 0°C
Preferably, the reaction is carried out by stirring at 120° C. for 1 hour to 2 days.

■Synthesis of compound α Compound α0 is the synthesis of compound (9) in the presence of a glycosidation catalyst in an inert gas atmosphere such as argon gas.
) can be obtained by reacting with Examples of the glycosidation catalyst include MeOTf, molecular sieve 4Δ, the aforementioned mercury-based catalysts, and silver thread catalysts. The reaction can also be carried out in a solvent such as dichloroethane, dichloromethane, nitromethane, acetonitrinobeatenobetoluene, THF, dioxane, etc.
It is preferably carried out by stirring for 5 minutes to 1 day at a temperature of 1 (10) °C to 1 (10) °C.

Compound (9) can be produced in accordance with Scheme 7 using compound (F) as a raw material via compound (G) as follows.

■Synthesis of Compound (G) Compound (G) can be obtained by reacting the easily available compound 4m(F) (fucostetraacetate) with methylthio(71-butyltin) in the presence of a catalyst such as tin tetrachloride. be able to. The reaction is preferably carried out in a solvent such as dichloroethane, chloroform, dichloromethinobe THE, etc. by stirring at -10°C to 80°C for 30 minutes to 1 day.

■Synthesis of compound (9) Compound (9) is produced by reacting compound (G) with an alkali methoxide such as NaOMe in a solvent such as methanol or ethanol at a temperature of -10°C to 80°C for 5 minutes to 1 day with stirring. It can be obtained by reacting the reaction product obtained by the reaction with sodium hydride and benzyl bromide. The reaction with sodium hydride and benzyl bromide is preferably carried out in a solvent such as THF, DMF, dioxane, toluene, etc. by stirring at a temperature of -10°C to 80°C for 5 minutes to 2 days.

■Synthesis of compound (11) Compound (n) is prepared by converting compound α into a solvent such as THF in the presence of an acid catalyst such as trifluoroacetic acid-water, acetic acid-water, or dilute hydrochloric acid.
It can be obtained by stirring at -10°C to 1(10)°C for 5 minutes to 2 days.

Below, compounds of the present invention 0, 03), 04), α6),
The manufacturing method for 0 is described according to schemes 3.4 and 5.

[Synthesis of compound 0] Compound a';! J is compound (11) with 10% Pd-C,
It can be obtained by hydrogen reduction in the presence of a catalyst such as 5% Pd-BaSO4, PtO2, or 5% Pt/C. The reaction is preferably carried out in a solvent such as methanol, ethanol, acetic acid, etc. by stirring at 0°C to 80°C for 1 hour to 4 days. After completion of the reaction, the catalyst is filtered off from the reaction mixture and the solvent is distilled off, followed by appropriate purification by known means such as gel filtration to obtain a pure product of Compound 0.

Note that the raw material for compound 03) does not require any particular purification.

[Synthesis of compound α3)] Compound 03) is prepared by diluting compound α in a solvent such as pyridine, dichloroethane, dichloromethane, THF, dioxane, etc.
- It can be obtained by reacting with acetic anhydride in the presence of DMAP. The reaction is preferably carried out by stirring at -10°C to 80°C for 30 minutes to 2 days. The resulting reaction mixture can be purified by known means such as column chromatography to obtain a pure compound α3).

[Synthesis of Compound 04) Compound 04) can be obtained by reacting Compound 0 with hydrazine acetate and then reacting the reaction product with diethylaminosulfur trifluoride (DAST). The reaction with hydrazine acetate is DMFXD
MS○, T) IF, in a solvent such as dioxane, -10°C ~
Preferably, the reaction is carried out under stirring at a temperature of 80° C. for 5 minutes to 10 hours. In addition, the reaction with DAST is carried out in a solvent such as dichloroethane, dichloromethane, THF, dioxane, toluene, etc., under an atmosphere of an inert gas such as argon gas, at -20°C to 6°C.
It is preferably carried out by stirring at 0°C for 5 minutes to 10 hours. After removing the solvent, the resulting reaction mixture is appropriately purified by known means such as column chromatography to obtain a pure compound α-Xun.

[Synthesis of compound 06) Compound 06) can be obtained by reacting compound 04) with ceramide compound α5) in the presence of a glycosidation catalyst. Examples of the glycosidation catalyst include Ag OTf and Snα2. The reaction is carried out in a solvent such as chloroform, dichloromethane, dichloroethane, toluene, THF, oxane, etc.
It is preferable to conduct the reaction with stirring at a temperature of -10°C to 60°C for 5 minutes to 1 day. After the reaction is completed, the solvent is removed from the reaction mixture, and the compound 06) can be purified as appropriate by known means such as column chromatography.

The ceramide compound α5) can be synthesized based on the description in JP-A-60-190745.

[Synthesis of Compound 0η] Compound 0η can be obtained by reacting compound α6) with an alkali methoxide such as sodium methoxide in a solvent such as methanol or ethanol. The reaction is preferably carried out at a temperature of -10°C to 80°C with stirring for 5 minutes to 1 day. By purifying the reaction mixture by known means such as gel filtration, compound α7) [Lewis b
type antigen] can be obtained.

〔Usefulness〕

The novel glycolipids of the present invention provide an effective means for elucidating the biological functions of abnormal carbohydrates due to cancerous changes in cells, and furthermore, monoclonal antibodies produced from the glycolipids can be used to diagnose various cancers. It is useful in that it can also be applied to testing and treatment.

The present invention will be explained in more detail below with reference to Reference Examples and Examples.

Reference Example 1 Synthesis of Compound (2) Compound (1) 17.6g (20mM), benzaldehyde dimethylacecool 9.1g (60mM) and p
-Toluenesulfonic acid 0.95 g (5 mM) was dissolved in 150 r+1 of tetrahydrofuran and stirred at room temperature overnight. After neutralizing the reaction solution by adding triethylamine, the tetrahydrofuran was concentrated under reduced pressure, and purified by column chromatography (developed with toluene/ethyl acetate - 5/1). was dissolved in 250 ml of tetrahydrofuran, and 13.2 g of borane-trimethylamine complex (BH3.Me, N) was dissolved in 250 ml of tetrahydrofuran.
(187mM) was added.

Under water cooling, add 24% of aluminum chloride powder to this reaction solution.
9g (187mM) was added little by little and stirred.

The reaction mixture was poured into ice water, extracted with ethyl acetate, washed successively with saturated sodium chloride water and saturated brine, dried over magnesium sulfate, the solvent was distilled off, and the oily residue was purified by column chromatography (toluene/acetic acid). The residue was purified with ethyl chloride (evolved with 5/1) to obtain 13.2 g (yield: 80%) of an oily substance.

Elemental analysis value C6 (H640HH・H20=991.1
Theoretical value as 99 C: 73.92 H: 6.48 Actual value
C: 73.85 H: 6.71 Optical rotation [α)o
-5,7° (C=1.52.CDα3)Rf=
0.50 (ethyl acetate/n-hexane:/=1/2)
'H-NMR (4(10)MHz, CDα3): δ7
．． 38-7.13 (m, 35H.

Ph), 4.49(d, IH, H-1b, J=
7.63Hz), 4.4Hd.

IH, H-1a, J=7.33) 13C-NMR (90MHz, CDα3): 6102
．． 454 (c, a, c, b) Reference Example 2 Synthesis of Compound (3) [Synthesis of Compound (B)] Compound (A) 4.8 g (10 mM) and methylthiotri-n-butyltin 3.8 g (, 10.5mM) was dissolved in 50ml of dichloroethane, 1.4ml of tin tetrachloride was added dropwise under ice cooling, and the mixture was stirred for 4 hours. After the reaction was completed, dichloroethane was distilled off under reduced pressure, and the residue was extracted with ethyl acetate.
Wash thoroughly with saturated potassium fluoride solution. The inorganic salts precipitated at this time were removed by suction filtration through Celite, the filtrate was washed with saturated saline, dried over magnesium sulfate, the solvent was distilled off, and 4.2 g of white crystals (yield 90%) were obtained. m, p, 145 -146℃.

Optical rotation [α], +49.7° (C=0.76, C
Dα3) Rf=0.40 (ethyl acetate/n-hex 7=1/1)'H-NMR (CDα3) δ near, 9-7.7 (m, 4H, aromatic).

5.40 (d, IH, H-1, J=10.3Hz).

2, 16(s, 3H, -3CH3), , 2.11
2.03.1.87 (3Xs, 3X3H.

3x [1Ac) [Synthesis of compound (C)] 107 g (0.23 M) of compound (B) was dissolved in methanol 1
1 and add 0. IN sodium methoxide 1
50mj7 was added and stirred at room temperature for 3 hours. Amberlyst 15 was added to the reaction solution and removed after neutralization, and the solvent was distilled off to give 71.2 g of white crystals (yield 91%) m, p, 16
>-B°C was obtained.

Of this, 6.78g (20mM) and 3.95g (26mM) of benzaldehyde dimethyl acecool
M) was dissolved in 80 ml of DMF, and p was added as an acid catalyst to this.
-Toluenesulfonic acid io Qmg was added and stirred overnight. The reaction solution was extracted with ethyl acetate, and saturated sodium hydroxide solution was extracted with ethyl acetate.
After sequentially washing with saturated saline and drying with magnesium sulfate,
The oil obtained by distilling off the solvent was purified by column chromatography (developed with ethyl acetate/n-hexane = 1/3) to obtain 6.82 g (yield: 82%) of an oil.

Optical rotation [α]. +2.0° (C=0.70.C
Dα3) Rf=0.67 (ethyl acetate/n-hext
= 1/1)'H-NMR (CD(J3) δ near, 9
-7.3 (m, 9H, aromatic).

5.56 (s, IH, Ph-CH).

5.28 ([3, IH, H-1, J・10.3Hz)
.

2,16 (s, 3H, -3CH3) [Compound (D
) Synthesis of Compound (C) 426mg (1mM) and Bora 7-)
438 mg (6 mM) of 'J methylamine complex was dissolved in 20 ml of tetrahydrofuran, and 798 mg (6 mM) of room temperature aluminum chloride powder was added little by little, followed by stirring for 2 hours. The reaction solution was poured into ice water, extracted with ethyl acetate,
After sequentially washing with saturated sodium chloride water and saturated brine, it was dried over magnesium sulfate, and the solvent was distilled off. The residue was purified by column chromatography (developed with ethyl acetate/toluene = 1/1) to obtain 320 mg (yield %) of an oily substance.

Optical rotation [α)D +4.5° (C=0.31.
CHα3)Rf=0.35 (ethyl acetate/toluene=2/1)'HNMR(CDα3)δ+7.8-7.2
(m, 9N, aromatic).

5.19 (d, IH, H-1, J=10.4Hz).

2.1NS, 3H, -3CH3) Elemental analysis value C2□H23N 06S=429.495
Theoretical value as C: 61.52 H: 5.40 N
: 3.26S near, 47 Actual value C: 61.06 H: 5.38 N: 3.
19S Near, 25 [Synthesis of compound (3)] 215 mg (0.5 mM) of compound (D) was dissolved in acetic anhydride/
It was dissolved in pyridine/4DMAP (1 ml/1 mβ/catalyst amount) and stirred at room temperature for 3 hours. Extract with ethyl acetate,
Washed with water, saturated soap water, diluted hydrochloric acid, and saturated saline, dried over magnesium sulfate, and distilled off the solvent.
mg (quantitative) m, p, 166-167°C was obtained.

Optical rotation CaE n +69.1@(C=0.87.
Ct(ci3)Rf=0.67 (ethyl acetate/
Toluene = 2/3)'It-NMR (CDC!3) δ near, 8-7.2 (m, 9H, aromatic).

5.37(d, 1)1. J’=10.4Hz).

2.17(S, 3H, -3[:H3).

1.91.1.86 (2XS, 2X3H.

2XOAc) Elemental analysis value C26H21N Os 5=513.57
Theoretical value C: 60.81. H:5.30
N: 2.73 S: 6.24 Actual value C: 60.67 H: 5.30 N: 2.
62S: 6.16 Reference Example 3 Synthesis of Compound (4) Compound (2) 78 g (8mM) and (3) 5.
2 g (10 mM) of dry nitromethane 1 (10)
ml and thoroughly dried in advance. M, 3.4A
The mixture was injected into a brown two-way flask containing 10 g under an argon gas stream.

Add 2.3 m of methyl triflate to this reaction solution at room temperature.
β (20mM) was added dropwise and stirred overnight.

The reaction mixture was filtered, and the filtrate was washed successively with water, saturated sodium chloride water, and saturated brine, dried over magnesium sulfate, and the solvent was distilled off under reduced pressure. The residual oil was dissolved in methanol-THF.
(Dissolved in 1(10)-20rr+j, this l:
0. I N Na OMe methanol solution IQrr
+j! was added and stirred at room temperature overnight. After neutralization by adding Amberlyst 15, the mixture was filtered off, and the filtrate was distilled off under reduced pressure.

The residual oil was purified by column chromatography (toluene/
Purified with ethyl acetate = 3/1 (M) to give an oily product of 8.5
4 g (yield 79%) was obtained.

Elemental analysis value C8□H,N O, = 1354.57, theoretical value C near 2.71 H: 6.18 Nil
, 03 actual value C near 2.51 H: 6.13 N
:1.15 optical rotation [α]. -27,9° (C=1.
85. ["Hα3)Rf=0.42 (ethyl acetate/toluene=1/1)'H-NMR (4(10) MI
(z, CDα3) δ7.36-6.9 (m, 44H
, Ph), 5.46 (d, IH.

H-1c, J=8.2tlz).

13C-NMR (90MHz, CDα3) δ102.
402 (Ca-, Cab) 99.745 (C, c') 55.264 (C2c) Reference Example 4 Synthesis of Compound (5) 3.02 g (2.23 mM) of Compound (4) at 2%
The mixture was dissolved in hydrazine-ethanol 1 (10) mβ and stirred under heating under reflux overnight.

Ethanol was distilled off under reduced pressure, and the residual oil was dissolved in methanol 1
(10) mβ, 1.5 ml of acetic anhydride was added dropwise thereto, and the mixture was stirred at room temperature for 1 hour.

Methanol was distilled off under reduced pressure, and the residue was extracted with ethyl acetate.
After washing with saturated brine, it was dried over magnesium sulfate, and the solvent was distilled off.

The residue was purified by column chromatography (developed with acetone/toluene=1/2) to give 2.08 g (yield 74%) of an oily substance.

Elemental analysis value C76H83N 016=1266, 50
Theoretical value as 6 C near 2.08 H: 6.61
N: 1.11 Actual value C near 2.01 H: 6.63
N: 1.08 optical rotation [αL -14,5°(
C=0.51. CHα3)Rf=0,32 (acetone/toluene=3/2)'H-NMR(4(10)
MHz, CDα3) δ7.37-7.10 (m, 4
0H, Ph), 5.52 (broadS.

LH, NH), 4.43 (d, IH, H-1b,
J=7.6Hz), 4.41(d, LH,H
-1a, J=7.9Hz).

1.37(s, 3H,HCO[:t13)”C-NM
R (90MHz, CDα3) δ102.510 (C,
,,C1b,C,c)58.569(C2c) Reference Example 5 Synthesis of Compound (6) 4.6-benzylidene derivative 9.36 g (30mM
) and borane-trimethylamine complex 13.1 g (1
80mM) was dissolved in 3 (10) ml of tetrahydrofuran, and 23.9 g (1
80 ml) was added little by little and stirred for 2 hours.

The reaction solution was poured into ice water, extracted with ethyl acetate, washed successively with saturated aqueous sodium bicarbonate and saturated brine, dried over magnesium sulfate, the solvent was distilled off, and the residue was subjected to column chromatography.
(Developed with acetone/toluene-1/1) to obtain 5.2 g (yield: 55%) of an oily substance.

This was mixed into acetic anhydride/pyridine/4-DMAP (5
mj! 15 mβ/catalyst amount) and stirred at room temperature overnight. The reaction solution was extracted with ethyl acetate, washed sequentially with water, saturated aqueous sodium bicarbonate, diluted hydrochloric acid, and saturated brine, dried over magnesium sulfate, the solvent was distilled off, and the resulting residue was subjected to column chromatography (ethyl acetate/ (Developed with toluene = 1/3).

Purified to give 7.2 g of white crystals (yield, quantitative), m, p.

53-54°C was obtained.

NMR (CDα3) δ: 7.3 (s, 5H, ar
omatic).

2, 72 (Q, 2H, -C8 February 1.' J=7.
5H2).

2, 05.・1.98 (3X S, 3X3H.

-DAC), 1.27(t, 3H, -CH2C
Flat.

J=7.5Hz) Optical rotation [αL −32,8” (c=o, 87.
CHα3)Rf=0.65 (ethyl acetate/toluene=1/2) Reference example 6 Synthesis of compound (7) Under an argon gas stream, compound (5N, 12 g (0
, 88mM) and compound (6) 790mg (1,8
mM) and dichloroethane-nitromethane (15-15
M, 3.4A 3.
g, 80 h+g (3.6 mM) of cupric bromide and 650 mg (1.8 mM) of mercuric bromide were poured into a brown double-necked flask, and the mixture was stirred at room temperature overnight.

The reaction mixture was filtered, and the filtrate was washed successively with saturated sodium chloride water and saturated brine, dried over magnesium sulfate, and the solvent was distilled off.

The residue was purified by column chromatography (developed with ethyl acetate/toluene=1/2) to obtain 1.11 g (yield: 77%) of an oil.

Optical rotation [α)n −9,2° (C=0.51.
CHα3)Rf=0.64 (ethyl acetate/toluene-3/2)'H-NMR (4(10)MHz, CDα
3) δ7.37-7.05 (m, 45H, Ph),
5.38(d, 1N.

H-4d, J=3.36Hz) 5.34(cl,
18. N.H.

J=3.05Hz) 2.05.1.97.1.95
(3XS.

3 x 31. 3X-0Ac)” CNMR (90MHz, CD C13) δ102
．． 454 (c, , c, b , c, c) 101° 154 (C, , ) 56.7.27 (C, c) Reference Example 7 Synthesis of Compound (8) Compound (7) 990 mg (0.6 mM ) was dissolved in methanol 15r+l, and Q, l N Na OM
2 ml of methanol solution was added, and the mixture was stirred at room temperature overnight. Neutralize by adding Amberlyst 15, remove by filtration, evaporate the filtrate under reduced pressure, and remove the oily residue with acetone-DMF (20 ml).
-2 ml) and add 190 ml of dimethoxypropane.
50 mg and p-)luenesulfonic acid were added, and the mixture was stirred under heating under reflux overnight. After neutralization by adding triethylamine, the solvent was distilled off, the residue was extracted with ethyl acetate, washed successively with water and saturated brine, dried over magnesium sulfate, and the solvent was distilled off under reduced pressure.

The residue was purified by column chromatography (developed with acetone/toluene-1/3) to obtain 660 mg (yield: 71%) of an oil.

Elemental analysis value C9□H+oJO2+ ・H20=157
Theoretical value as 6.855 C near 0.08 H: 6.71 N: 0.
89 Actual value C near 0.05 H: 6.62 N
:0.85 optical rotation [αL -7,8° (C=1.3
6. CHα3)Rr=0.56 (acetone/toluene=1/2)' HNMR (4(10)MHz, CD
α3) δ near, 40-7.10 (m, 45H, Ph
).

2, 03. 1.96 (2XS, 2X3H, 2x-
OAc) 1, 30 (s, 3) 1. NAc)13
C-NMR (90MHz, CDα3) δ: 102.
345 (Ca-, Cab) 55.644 (C2c
) 103.919 (C, c) 101.643 (C, d) Reference example 8 Synthesis of compound (9) 3.3 g (1
0mM) and 3.8g methylthiotri n-butyltin
(10.5mM) was dissolved in 40rr+j2 of dichloroethane, 1.4ml of tin tetrachloride was added dropwise under water cooling, and the mixture was stirred for 4 hours. After the reaction is complete, dichloroethane is distilled off under reduced pressure, and the residual oil is extracted with ethyl acetate and thoroughly washed with saturated potassium fluoride solution. At this time, the precipitated inorganic salt was removed by suction filtration through Celite, and the filtrate was washed with saturated brine, dried over magnesium sulfate, and the solvent was distilled off. The residue was purified by column chromatography (developed with ethyl acetate/n-hexane = 1/2) to obtain 1.1 g of α-thiomethyl derivative (yield 34%) and 1.8 g of β-thiomethyl derivative (yield 56%).

α-thiomethyl derivative m, p, 80-81°C Optical rotation [α], −222° (C = 1.2. CH
α3)'II-NMR (CDα3)δ: 2.16.
2.07. 2.05. 1.99 (4XS, 4X
3H, 4XOAc) 1, 17 (d, 3H, -CL, J=6.6Hz)
Optical rotation of β-thiomethyl derivative m, p, 139-141 [α], -0,7° (C = 1.0. CH
α3)'H-N), IR(CD(J3)δ: 4.35
(d, 18. H-1, J=9.4)1z)2.19
．． 2.17. 2.07. 1.99 (4XS, 4
X3H, 4XOAc) 1, 22 (d, 3H, -CL, J=6.6Hz)
27.2 g (85 mM) of the β-thiomethyl derivative was dissolved in 3 (10) ml of methanol, and 0.2 g (85 mM) of the β-thiomethyl derivative was dissolved in 3 (10) ml of methanol. IN sodium methoxide 50rr+jl! Add and at room temperature,
Stirred for 4 hours. Amberlyst 15 was added to the reaction solution to neutralize it, filtered off, and the solvent was distilled off to give 17.4 g of white crystals.
(Quantitative) m, I], 103-104°C was obtained.

Of this, 15.5g (80mM) was added to 60%
A suspension of 16 g (320 mM) of sodium hydride in 2 (10) ml of DMF was added little by little at room temperature and stirred for 30 minutes. Next, 38 ml of benzyl bromide (32
0mM) and stirred overnight. The reaction solution was poured into ice water, extracted with ethyl acetate, washed successively with dilute hydrochloric acid and saturated brine, dried over magnesium sulfate, and the solvent was distilled off. The residual oil was purified by column chromatography (ethyl acetate/n
-Developed with hexane = 1/10), and purified with oily substance 33
．． 5 g (yield 90%) was obtained.

Optical rotation [α]DO, 2° (C=1.54.
CHCl3)Rf=0.50 (ethyl acetate/n-hex/=1/4)'H-NMR (CDα3)δ: 7.
5-7.2 (m, 15H, aromatic) 1, 2
0 (d, 3H, -CL, J=6.4Hz) Reference Example 9 Synthesis of compound αQ Compound (8) 441mg (0,282mM) and (
9) 395mg (0.85mM) was dissolved in dry nitromethane 30mA and thoroughly dried beforehand), 1. S,
A brown double-necked flask containing 3 g of 4A was injected under a stream of argon gas.

Methyl triflate 2 (10
) μ! (1.7 m old was added dropwise and stirred for 2 hours.

The reaction mixture was filtered, and the filtrate was washed successively with water, saturated sodium chloride water, and saturated brine, dried over magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue was subjected to column chromatography (
The residue was purified using ethyl acetate/toluene (1/6 development) to obtain 402 mg (yield: 60%) of an oily substance.

Elemental analysis value Cl468I59N○2゜・H2O-24
Theoretical value as 09,899 C near 2.77 H: 6.73 N: 0.
5g actual value C near 2.39 H: 6.63 N
:0.59 optical rotation [α]. -37,0° (C・0.8
6. CDα3)Rf=0.54 (ethyl acetate/
Toluene = 1/3)'H-NMR (4(10)MHz,
CDC13) C7,36-7,07(m, 75H,
Ph), 5.38 (d, LH.

H-1f, J=3.67Hz) 5.09 (d,
IH, H-Le.

J=3.66Hz) 1.37.1.34. (2XS
, 2x3H.

1sopropylidene rnetyl),
1.14.1.11 (2Xd, 2X3H, 2XFu
c-CL, J=6.71Hz) "C-NMR (90MHz, CDcJl,) δ:1
02.345(C,,,C,b) 101.804
(C, c) 101.262 (C,,) 97.30
9 (C,.) 95.410 ([:, f) 59.1
11(C2c)27.849. 26.332 (isopropylidene-CH,) 22.972 (NAc) 16
．． 904. 16.525((5))Fue(-CH,
) Reference Example 10 Synthesis of Compound Ql) Compound αG 325mg <0.208mM) at 80%
It was dissolved in 10 mf of trifluoroacetic acid/THF (1/1) and stirred at room temperature overnight.

The mixture was diluted and extracted with ethyl acetate, washed successively with saturated sodium chloride water and saturated brine, dried over magnesium sulfate, and the solvent was distilled off. The residue was purified by column chromatography (ethyl acetate/
Purified with toluene-1/2 development) to obtain 320mg of oily substance.
(quantitative).

Elemental analysis value Cl43H155N029=2351.8
Theoretical value as 19 C near 3.03 H: 6.64
N: 0.60 Actual value C near 2.85 H: 6.
62 N: 0.59 optical rotation [α] knee-33,6
°(C=0.66, CDα3)Rf=0.48
(Ethyl acetate/toluene-1/2)' HN! JR (4
(10)! Jtlz, CDCf 3) δ near, 4-
7.05 (m, 75H, Ph), 5.22 (d
, L.H.

H-1f, J=3.66Hz) 5.05 (
d, IH, H-1e.

J=3.66Hz), 1.43(s, 3)1.
NAc).

1.15. 1.11. (2Xd, 2X3H,
2XFuc (-CH3), J=6.71Hz) 13C-NMR (90MHz, CDα3) δ: 10
2.345(C,,,C,b,C,c)1(10
). 828(C,,) 99.801(C,,)9
7.469(C,,) 59.219(C2e)2
3. (10)0(NAc) 16.904. 16.
579 (Fuc(-CH3)) Example 1 Synthesis of compounds α and θ 310 mg (0,131 ml 1 J) of compound (11) was dissolved in methanol-acetic acid (3-3 rTW), and 1
Add 0% Pd-C1 (10) mg, under hydrogen gas flow,
Stirred for 4 days. The Pd-C was filtered out, the filtrate was distilled off under reduced pressure, and 15 mg of the resulting residue was filtered using the gel filtration method (G-2
5) to obtain compound α. The remaining residue was dissolved in acetic anhydride/pyridine/4-dimethylaminopyridine (1 m
j! /1 mβ/catalyst amount) and stirred at room temperature overnight. The extract was extracted with ethyl acetate, washed successively with water, saturated sodium chloride water, diluted hydrochloric acid, and saturated brine, dried over magnesium sulfate, and the solvent was distilled off. The residue was purified by column chromatography (developed with acetone/toluene-1/3) to obtain 182 mg (yield: 91%) of an oil.

Compound (121 Optical rotation [αL-44,3° (C=0.56.820
)Rh0.22 (n-butanol/ethanol/water=2/1/1) 'H-NMR (4(10)MH2,D20) δ:5
．． 20(d, LH, It-1, α, , J=3.9
6Hz) 5, 13 (d, ltl, H-1r, ,
J'3.661(z).

5.01 (d, LH, H-1, J=3.66Hz)
.

4.64 (2xd, 2H, HL, HL, J=
7.63H2), 4.58(ill, LH,H-
LH, J=8.55Hz), 4.40(d, l
tl, H-1c, 2.04(s, 3H,N
Ac), , 1.26 (2xd, 2x3H.

Fuc (CH3) J=6.41Hz) Compound 0→ Rf=0.33.0.36 (acetone-toluene=
1/1)'H-NMR (4(10)MHz, CDCl
1Jδ: 5.42 (d, LH, H-1f) 5.1
3 (d, IH, H-1e) 4.51 (LH, d, H-
1d, J=7.63Hz), 4.34(d, L
H, H-1a, J=7.93), 2.2-1.9
2 (multi S, 54H, Ac-), 1.2
0d, 6H.

FUC(CH3)) Example 2 Synthesis of Compound 04) Compound Q3) 171 mg (0.1 mM) was DM
3 ml of F was dissolved, 18 mg (0.2 mM) of hydrazine acetate was added thereto, and the mixture was stirred at room temperature for 30 minutes.

The reaction solution was diluted with ethyl acetate, washed sequentially with water and saturated brine, dried over magnesium sulfate, the solvent was distilled off, and the residue was subjected to column chromatography (acetone/toluene=
3/7) to obtain 110 mg (yield: 65%) of an oily substance.

This material was mixed with dichloroecune/THF under an argon atmosphere.
(4/1 ml), 35 μl of diethylaminosulfur trifluoride (DAST) was added thereto under ice cooling, and the mixture was stirred for 30 minutes. The reaction solution was poured into cold saturated sodium hydrogen chloride water and extracted with chloroform. This was washed with saturated brine, dried over magnesium sulfate, the solvent was distilled off, and the residue was subjected to column chromatography (acetone/
Purified with toluene = 1/2 development) to obtain 57 mg of oil (
A yield of 52% was obtained.

Rf=0.47 (acetone-toluene=1/1)'
H-NMR (4(10)MHz, C[]α,)δ
:5.41(d, 1)1. H-1f, J=3.
36)1z).

5.12 (d, 18. H-1e, J=3.6
6Hz).

4.51 (d, IH, H-1d, J=7.63
).

4.26 (d, LH, H-1aβ, J□8.5
5Hz).

2.18-1.92 (multi S, 5114
．． -8C).

1.21 (d, 6H, 2xFuc(-CH,)) Example 3 Synthesis of compound α6) Under argon gas flow, compound α4) 52 mg (0,
031mM) and α5) 45mg (0.06mM)
was dissolved in chloroform/toluene (2/2ml),
Dry thoroughly beforehand. M, S, 4A 2(10)TI
1g, silver triflate 16mg (0.06mM
) and 12 mg (0.06 mM) of stannous chloride into a brown double-necked flask, and the mixture was stirred at room temperature for 4 hours.

The reaction mixture was filtered, and the filtrate was washed successively with saturated sodium chloride water and saturated brine, dried over magnesium sulfate, and the solvent was distilled off. The residue was subjected to column chromatography (toluene/
It was purified by developing with chloroform/acetone=2/1/1 to obtain 6.2 mg (yield 9%) of an oily substance.

Optical rotation [αlfl -46,9° (C=0.29.
Cllα3)Rf・Q, 50 (acetone/toluene=2/3)'II-NMR (4(10)MHz,
CDα3)δ:5.87(m, IH,H-5Cer
).

5.73 (m, LH, fl-4Cer).

5.40 (d, lft, tl-1f, J=3.
36Hz).

5, 12 (d, IH, H-1e, J=3.97H
z).

4.51 (d, LH, H-1d).

4.43 <d, IH, o-td, J=7.
63);4.30 (d, IH,1(-1a,
J=7.93).

2.17-1.92 (multi S, 51H, -
Ac).

0.87 (t, 6H, 2xCer(-Ctf, )) Example 4 Synthesis of compound αη 1 (10) mf of NNaOMe methanol solution was added and stirred overnight.

Amberlyst 15 was added to the reaction solution to make it neutral, then filtered off, and the filtrate was distilled off under reduced pressure. Gel filtrate the residue (L
H-2'0) (chloroform/methanol/water = 60/
Compound Q? ) 3. , 8 mg (yield q.

%) was obtained.

Optical rotation [αL, 42.1° (C=0.19.
CDα3) Rf=0.52 (n-butanol/ethanol/water=2/1/1) 'H-NMR (4(10)MHz, DMSO-d6)
δ: 5.55 (m, IH,)I-5Cer).

5.35 (m, IH, H-4Cer).

4.90 (d, IH, tl-1f, J=3.3
6Hz).

4.78 (d, IH, H-1e), 4.66 (q,
LH, H-5e).

4.58 (d, IH, H-1c, J=8.34
Hz).

4.54 (d, IH, H-1d, J=7.02
Hz).

4.28 (d, LH, H-1b), 4.21 (q,
IH,H-5f)4.18(d,LH,)l-
1a, J=7.93Hz).

1.85 (s, 3H, NAc), 1.15,1
．． 12 (2Xd.

2X3H, 2xFuc(CH3)).

Claims (11)

[Claims] (1) A compound represented by the general formula [I]. ▲There are mathematical formulas, chemical formulas, tables, etc.▼[I] (In the formula, R^1 is a hydrogen atom or an acetyl group, and R^
2 is a hydroxyl group, an acetyloxy group, a fluorine atom, or a ▲ mathematical formula, chemical formula, table, etc. ▼ group (however, R^3 is a hydrogen atom or a benzoyl group), and Ac represents an acetyl group. ) (2) Formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (In the formula, Bn represents a benzyl group and Ac represents an acetyl group.) From hydrogen reduction of the compound (11) in the presence of a catalyst. A method for producing a compound (12) represented by the formula ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (In the formula, Ac represents an acetyl group.) (3) The catalyst is Pd-C, Pd-BaSO_4, PtO_
2 or Pt-C. (4) By reacting the compound (12) represented by the general formula [I] (where R^1 is a hydrogen atom and R^2 is a hydroxyl group) with acetic anhydride in the presence of 4-DMAP. A method for producing a compound (13) represented by the general formula [I] (wherein R^1 is an acetyl group and R^2 is an acetoxy group). ▲There are mathematical formulas, chemical formulas, tables, etc.▼〔I〕 (5) The production method according to claim (4), in which pyridine, dichloroethane, dichloromethane, THF, or dioxane is used as the reaction solvent. (6) Compound (13) represented by general formula [I] (wherein R^1 is an acetyl group and R^2 is an acetoxy group) is reacted with hydrazine acetate, and then diethylaminosulfur trifluoride Compound (14) represented by the general formula [I] consisting of reacting with
) (wherein R^1 is an acetyl group and R^2 is a fluorine atom). ▲There are mathematical formulas, chemical formulas, tables, etc.▼〔I〕 (7) DMF as a solvent for the reaction with hydrazine acetate;
The manufacturing method according to claim 6, wherein DMSO, THF or dioxane is used and dichloroethane, dichloromethane, THF, dioxane or toluene is used as a solvent for the reaction with diethylaminosulfur trifluoride. (8) Compound (14) represented by the general formula [I] (where R^1 is an acetyl group and R^2 is a fluorine atom) in the presence of a catalyst ▲ where the mathematical formula, chemical formula, table, etc. There is a compound (16) represented by the general formula [I] which is obtained by reacting with ▼ (in the formula, Bz represents a benzoyl group).
However, R^1 is an acetyl group, and R^2 is ▲ formula,
There are chemical formulas, tables, etc. It is a ▼ group (in the formula, Bz represents a benzoyl group). ) manufacturing method. ▲There are mathematical formulas, chemical formulas, tables, etc.▼〔I〕 (9) The production method according to claim (8), wherein the catalyst is silver chloride and silver trifluorate, and the reaction solvent is chloroform, dichloroethane, dichloromethane, toluene, THF, or dioxane. (10) Compound (16) represented by general formula [I] (
However, R^1 is an acetyl group, and R^2 is a ▲ mathematical formula, chemical formula, table, etc. ▼ group (in the formula, Bz represents a benzoyl group). ) is reacted with alkali methoxide to form a compound (17) represented by the general formula [I] (where R^1 is a hydrogen atom and R^2
▲There are mathematical formulas, chemical formulas, tables, etc. ▼The manufacturing method of the group). ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ (11) The production method according to claim (10), in which methanol or ethanol is used as the reaction solvent.