JPH0819161B2 - Novel glycosphingolipid-related compounds - Google Patents

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel fucose-substituted lactoseries glycosphingolipid-related compound.

2. Description of the Related Art SSEA-1 antigen (Stage-specific Embryonic Antigen), which is one of the differentiation antigens, is 1978.
Year, Solter and Knowles, etc. (Dee Salter and BB Knowles: Proc. Natl.
Acad. Sci. USA., 75, 5565-5569, 197.
8) is an antigen recognized by a monoclonal antibody obtained by immunizing a syngeneic mouse with mouse tetracarcinoma cell F9. Its structure is Le as an antigenic determinant at the non-reducing end.
^The fucose-substituted lactoseries glycosphingolipid having an ^X structure was reported in 1982 by Hakomori et al. Kannagi, B. Nudelman, SB Levery
and S. Hakomori): J. Biol. Chem., 257, 148.
65-14878, 1982; Earl. R. Kannagi, E. Nuderman and R. Kannagi, E. Nu
delman, and S. Hakomori): Proc. Natl. Acad. Sci.
USA., 79, 3470-3474, 1982],
Was revealed.

These are not initially thought of as a single antigen molecule, but they form a wide variety of sugar chain groups depending on the extent of fucosylation and the active extension of the N-acetyllactosamine structure, resulting in various cancers. It has been isolated from tissues as a cancer-associated glycolipid. [H. Jay Young and S.
Hakomori (HJ Yang and S. Hakomori): J. Biol.Ch
em., 246, 1192, 1971; S. Hakomori,
S. Hakomori, E. Nudelman, SB Levery
and R. Kannagi): J. Biol. Chem., 259, 467.
2, 1982; Fukushi Wai, S. Hakomori, E.
Nudermann and NA Cochilan (Y. Fukushi,
S. Hakomori, E. Nudelman and NA Cochran): 25
9, 4681-4685, 1984].

The present inventors have found that the glycolipid (Le ^x ) Galβ1 → 4 [Fucα1 → 3] GlcNAcβ1 → 3Galβ1 → 4Gl consisting of five sugar residues, which can be said to be the basic structure of the SSEA-1 antigen group.
Successful synthesis of cβ1 → Cer.

[0004]

THE INVENTION Problems to be Solved] In addition, appears to be closely related to the canceration in order, glycolipids of 8 sugar residues having a Le ^x repeated as structure (2Le ^x) Galβ1 → 4 [Fucα1 → 3] GlcNAcβ1 → Galβ1 → 4
Precise synthesis of [Fucα1 → 3] GlcNAcβ1 → 3Galβ1 → 4Glcβ1 → Cer is indispensable for promoting the elucidation of the biochemical role of these cancer-related sugar chains from the viewpoint of organic chemistry, and these glycolipids. Is significant as it is expected to be applied to diagnosis and treatment in the future. An object of the present invention is to provide a glycosphingolipid-related compound which is an important intermediate in the synthesis of a glycosphingolipid-related compound 2Le ^x consisting of 8 sugar residues.

[0005]

The present invention has the general formula [I]
The present invention provides a glycosphingolipid-related compound represented by

Embedded image In the formula, R ₁ , R ₂ and R ₃ are independently a hydrogen atom, a benzyl group or an acetyl group, R ₄ is a phthaloylamino group or an acetylamino group, R ₅ is a benzyl group,
An acetyl group or a -C (= NH) -CCl ₃ group. The above compound of the present invention can be produced, for example, by the following steps (a) to (d). (a) A step of reacting the compound [15] and the compound [16] to produce the compound [17]. (b) A step of treating compound [17] with hydrazine to eliminate the phthaloyl group and then acetylating to produce compound [19]. (c) A step of producing a compound [20] by debenzylating the compound [19] and then acetylating it. (d) After treating the compound [20] with hydrazine / acetic acid,
A step of producing compound [21] by treating with trichloroacetonitrile.

[0006]

Embedded image

[0007]

[Chemical 4]

[0008]

Embedded image

[0009]

[Chemical 6]

[0010]

[Chemical 7]

[0011]

Embedded image

Octasaccharide compound (2Le ^x ) which is the final compound
Can be produced, for example, by steps (e) and (f). (e) Compound [21] is reacted with ceramide compound [22] to give compound [23]. (f) The protecting group of compound [23] is removed to produce compound [1], that is, 2Le ^x .

[0013]

[Chemical 9]

[0014]

[Chemical 10]

[0015]

[Chemical 11]

In this specification, Ac represents an acetyl group, Bn represents a benzyl group, Ph represents a phenyl group, and Phth represents a phthaloyl group. Hereinafter, the method for producing the glycosphingolipid-related compound of the present invention will be described in more detail. First known compound (2), NaOCH _3, NaOEt, Na ₂ CO _3, NaHCO presence of a base such as _3, methanol, ethanol and the like in a solvent -1
It is deacetylated by treating at 0 ° C to 80 ° C for 5 minutes to 1 day to obtain a novel compound [3]. Then, in a solvent such as DMF (dimethylformamide), acetone, THF (tetrahydrofuran), toluene or dioxane, the compound [3] is converted to dimethoxypropane in the presence of toluenesulfonic acid, benzenesulfonic acid, anhydrous hydrochloric acid or zinc chloride. A novel compound [4] is obtained by reacting at 0 ° C to 120 ° C for 1 hour to 2 days for isopropylidene formation.
At this time, a novel compound [5] is also obtained as a by-product. Then, the compound [4] is reacted with benzyl bromide, silver oxide and potassium iodide at −20 to 100 ° C. for 5 minutes to 1 day in a solvent such as DMF, THF or dioxane to benzylate the novel compound [6]. ]] Is obtained. Furthermore,
The compound [6] is treated with trifluoroacetic acid / water, acetic acid / water, dilute hydrochloric acid or the like at −10 ° to 100 ° C. for 5 minutes to 1 day to eliminate the isopropylidene group to obtain a novel compound [8]. At this time, a novel compound [7] from which fucose has been eliminated is also produced as a by-product.

[0017]

[Chemical 12]

[0018]

[Chemical 13]

[0019]

Embedded image

Then, the compound [8] is heated in a solvent such as pyridine, dichloroethane, dichloromethane, chloroform, THF or dioxane at a temperature of -10 to 80 ° C for 5 minutes to
A novel compound that is acetylated by reacting with acetic anhydride for 2 days

Obtain [9]. In addition, the compound in a solvent such as acetic acid aqueous solution, methanol or ethanol

Compound [10] is obtained by treating [9] with palladium chloride at 0 to 120 ° C. for 30 minutes to 2 days. Then, in a solvent such as chloroform, dichloromethane, dichloroethane, toluene or THF in the presence of DBU (diazabicycloundecane), -2
The compound [10] is reacted with trichloroacetonitrile at 0 to 60 ° C. for 5 minutes to 1 day to give a novel compound [11].
Get. In a solvent such as chloroform, dichloromethane, dichloroethane, toluene or THF, BF ₃ · E
Compound [11] in the presence of a glycicylation catalyst such as t ₂ O or TMSOTf (trimethylsilyl triflate)
Then, the compound [12] is reacted to obtain a novel compound [13] having a pentasaccharide residue. Compound [14] as a by-product
Can also be obtained.

[0021]

[Chemical 15]

[0022]

Embedded image

[0023]

[Chemical 17]

Then, the compound [13] is treated with NaOCH ₃ , NaOC ₂ H ₅ , Na ₂ CO ₃ or NaHCO _{3 in} a solvent such as methanol or ethanol at −10 to 80 ° C. for 5 minutes to 1 day. Deacetylation gives a new compound [15]. Further, in a solvent such as chloroform, dichloromethane, dichloroethane, toluene or THF, BF ₃ · E
In the presence of a catalyst such as t ₂ O or TMSOTf, the compound [1
The known compound [16] in 5] is reacted at −20 to 60 ° C. for 5 minutes to 1 day to obtain a novel compound [17] and a novel compound [18] as a by-product.

[0025]

Embedded image

Further, the compound [17] is treated with hydrazine in a solvent such as methanol or ethanol at 0 to 120 ° C. for 5 minutes to 2 days to eliminate the phthaloyl group, and then pyridine, chloroform, dichloroethane, dichloromethane or THF. In a solvent of 10 to 80 ° C. for 30 minutes to 2
For a day, if necessary, it is reacted with acetic anhydride in the presence of 4-DMAP (4-dimethylaminopyridine) to acetylate to obtain a novel compound [19]. Then, in a solvent such as methanol, ethanol or acetic acid, 10% Pd-C, 5% Pd-
Compound [19] in the presence of a catalyst such as BaSO ₄ and 5% Pt-C
Is dehydrogenated by hydrogen catalytic reduction at 0 to 100 ° C. for 1 hour to 3 days, and then acetic anhydride, pyridine and 4-DMAP are added to -10 to 10 in a solvent such as pyridine, chloroform, dichloroethane, dichloromethane or THF. Acetylation is performed by reacting at 80 ° C. for 30 minutes to 2 days to obtain a novel compound [20] (yield 78%). Then DMF, DM
Compound [20] is treated in the presence of hydrazine / acetic acid in a solvent such as SO, THF or dioxysan at -10 to 100 ° C for 5 minutes to 1 day to remove the acetyl group at the 1-position, and then chloroform, dichloromethane or dichloroethane. ,
In a solvent such as toluene or THF in the presence of DBU,
Compound [21] is obtained by reacting with trichloroacetonitrile at 20 ° C to 60 ° C for 5 minutes to 1 day. Then, in a solvent such as chloroform, dichloromethane, dichloroethane, toluene or THF, BF ₃ · Et ₂ O and TMS are added.
The compound [21] is reacted with the ceramide compound [22] at −20 to 60 ° C. for 5 minutes to 1 day in the presence of a catalyst such as OTf to obtain a novel compound [23]. At this time, a novel compound [24] is obtained as a by-product.

[0027]

[Chemical 19]

[0028] Then n-Bu ₄ NF presence in a solvent such as THF or dioxane, compound [23] -20～10
After reacting at 0 ° C. for 5 minutes to 2 days, the reaction mixture is added with a solvent such as methanol, ethanol or THF to obtain NaOCH ₃ , NaOC.
With a base such as ₂ H ₅ , Na ₂ CO ₃ or NaHCO ₃ , -10 to 8
The target compound [1] (2Le ^x ) is obtained by removing the acetyl group by treatment at 0 ° C. for 5 minutes to 1 day.

[0029]

The glycosphingolipid-related compound of the present invention is an octasaccharide compound essential for promoting the elucidation of the biochemical role of cancer-related sugar chains from the viewpoint of organic chemistry.
It is an important synthetic intermediate for Le ^x . This 2 Le ^x
Is a glycolipid consisting of octasaccharide residues which is a basic structure of SSEA-1 antigen group, and anti-SS formed using this as a antigen
The EA-1 antibody can be applied as a serum tumor marker in the diagnosis and treatment of lung adenocarcinoma, ovarian cancer, pancreatic cancer and the like.
Moreover, 2Le ^x can be produced in good yield by using the glycosphingolipid-related compound of the present invention.

The present invention will be described in more detail with reference to Reference Examples and Examples. [Reference Example 1] (Synthesis of Compound [3]) Compound [2] (5.62 g, 4.73 mM) was added to methanol / T.
Dissolve in HF (20ml / 20ml) and add 0.2N NaO
5 ml of CH ₃ methanol solution was added and stirred at room temperature for 1 hour. After completion of the reaction, Amberlyst 15 was added to neutralize the reaction solution, which was then filtered off, and the filtrate was evaporated under reduced pressure. The residue was purified by column chromatography (acetone / toluene = 1/1 development) to obtain 4.82 g (yield, quantitative) of an oil. Rf = 0.46 (toluene / acetone = 1/1) [α] ^{_{D 25 -19.9 ° (C = 0.88}} , CHCl 3) Elemental _{analysis: C 57 H 63 NO 16 ·} 1 / 2H 2 O = 1027.141 calculated value C; 66.65 H; 6.28 N; 1.36 measured value C; 66.68 H; 6.27 N; 1.34 ¹ H-NMR (400 MHz, CDC1 ₃ ) δ; 7.75-6.9
8 (m, 24H, aromatic H), 5.736 (m, 1H, -CH ₂ - CH = C
H ₂ ), 5.219 (d, 1H, H-1a, J = 8.8Hz), 5.145 (dd, 1H, = CH
₂ , J = 1.5, 17.4HZ), 5.061 (dd, 1H, = CH ₂ , J = 1.5, 1
0.8Hz), 4.873 (d, 1H, H-1c, J = 2.7Hz), 4.489 (d, 1
H, H-1b, J = 7.0 Hz), 1.052 (d, 3H, H-6c, J = 6.4 Hz) ¹³ C-NMR (CDCl ₃ ) δ; 168.230 ( C ═O), 99.85
3 (C-1b), 98.661 (C-1a), 96.984 (C-1c), 55.860 (C-2a),
16.362 (C-6c)

Reference Example 2 (Synthesis of Compounds [4] and [5]) Compound [3] (4.58 g, 4.5 mM) was dissolved in DMF (20 ml), and dimethoxypropane (4.68 g, 45 m) was added.
M) and p-TsOH (300 mg) were added, and the mixture was stirred at room temperature for 2 days. After the reaction is completed, neutralize by adding triethylamine,
The mixture was extracted with ethyl acetate, washed successively with water and saturated brine, dried over magnesium sulfate, and the solvent was evaporated. The residue was dissolved in 50 ml of methanol, Amberlyst 15 was added thereto, the mixture was stirred at room temperature for 30 minutes, filtered, and the filtrate was evaporated under reduced pressure. The residue was purified by column chromatography (toluene / acetone = 5/1 development) and compound [4] 3.52 g
(Yield 73.9%) and Compound [5] 0.77 g (Yield 1
6.2 g) was obtained.

Compound [4] Rf = 0.65 (toluene / acetone = 2/1) [α] _D ²⁵ −7.0 ° (C = 0.79, CHCl ₃ ) Elemental analysis value: C ₆₀ H ₆₇ NO Calculated as ₁₆ = 1058.199 C; 68.10 H; 6.38 N; 1.32 Measured C; 67.92 H; 6.43 N; 1.27 ¹ H-NMR (400 MHz, CDCl ₃ ) δ; 7.73-6.96
(m, 24H, aromatic H), 5.727 (m, 1H, -CH ₂ - CH ＝ C
H ₂ ), 5.195 (d, 1H, H-1a, J = 8.5Hz), 5.140 (dd, 1H, =
CH ₂ , J = 1.5, 17.4Hz), 5.055 (dd, 1H, = CH ₂ , J = 1.5,
10.4Hz), 4.784 (d, 1H, H-1c, J = 3.4Hz), 4.396 (d,
1H, H-1b, J = 8.2Hz), 1.469, 1.313 (2 × s, 6H, = C
(CH ₃ ) ₂ ), 1.052 (d, 3H, H-6c, J = 6.4 Hz) ¹³ C-NMR (CDCl ₃ ) δ; 168.178 (C═O), 110.20
4 ( = C (CH ₃ ) ₂ ), 99.476 (C-1b), 98.718 (C-1a), 96.984 (C
-1c), 55.914 (C-2a), 28.119, 26.277 (C (CH ₃ ) ₂ ), 16.525
(C-6c)

Compound [5] Rf = 0.43 (toluene / acetone = 2/1) [α] _D ²⁴ + 2.5 ° (C = 0.56, CHCl ₃ ) Elemental analysis value: C ₆₀ H ₆₇ NO ₁₆ · 1 / 2H ₂ O = 1067.206 calculated C; 67.53 H; 6.42 N; 1.31 measurements C; 67.49 H; 6.39 N; 1.21 1 H-NMR (400MHz, CDCl 3) δ; 7.72-7.02
(m, 24H, aromatic H), 5.649 (m, 1H, -CH ₂ - CH ＝ C
H ₂ ), 5.113 (d, 1H, H-1a, J = 8.5Hz), 5.070 (dd, 1H,
= CH ₂ , J = 1.5, 17.1Hz), 4.983 (dd, 1H, = CH ₂ , J = 1.
5, 10.4Hz), 4.833 (d, 1H, H-1c, J = 3.7Hz), 4.551
(d, 1H, H-1b, J = 7.6Hz), 1.392, 1.301 (2 × s, 6H,
= C (CH ₃ ) ₂ ), 1.098 (d, 3H, H-6c, J = 6.4 Hz) ¹³ C-NMR (CDCl ₃ ) δ; 168.013 (C = O), 101.426
(C-1b), 98.718 ( = C (CH ₃ ) ₂ ), 97.686 (C-1a), 97.469 (C
-1c), 56.402 (C-2a), 29.474, 18.475 (C (CH ₃ ) ₂ ), 16.687
(C-6c)

[Reference Example 3] (Synthesis of Compound [6]) In an argon gas atmosphere, a flask containing 3.01 g (13 mM) of silver oxide was charged with Compound [4] 2.65 g (2.5 mM).
And 4.27 g (25 mM) of benzyl bromide are dried D
It was dissolved in 25 ml of MF and injected. 2.16 g (13 mM) of potassium iodide was added to this reaction solution under ice cooling, and the mixture was stirred for 1 hour. After completion of the reaction, the reaction solution was diluted with ethyl acetate, the insoluble matter was filtered off from Celite, and the filtrate was diluted with water, diluted hydrochloric acid,
The organic layer was washed successively with saturated brine and dried over magnesium sulfate, and the solvent was distilled off. The residue was purified by column chromatography (toluene / ethyl acetate = 7/1 development) to obtain 2.72 g (yield, 87.9%) of an oily substance. Rf = 0.59 (toluene / ethyl acetate = 6/1) [α] _D ²⁷ −2.2 ° (C = 0.87, CHCl ₃ ) Elemental analysis value: Calculated as C ₇₄ H ₇₉ NO ₁₆ = 1238.452 C; 71.77 H; 6.43 N; 1.13 Measured value C; 71.80 H; 6.43 N; 1.13 ¹ H-NMR (400 MHz, CDCl ₃ ) δ; 7.72-7.
01 (m, 34H, aromatic H), 5.647 (m, 1H, -CH ₂ - CH = CH
₂ ), 5.134 (d, 1H, H-1a, J = 8.5Hz), 5.069 (dd, 1H, =
CH ₂ , J = 1.5, 17.1Hz), 4.983 (dd, 1H, = CH ₂ , J = 1.5, 1
0.4Hz), 4.805 (d, 1H, H-1c, J = 3.2Hz), 4.155 (t, 1
H, H-2a, J = 9.5Hz), 3.528 (d, 1H, H-4c, J = 1.5Hz), 1.
321, 1.293 (2 × s, 6H, = C (CH ₃ ) ₂ ), 0.990 (d, 3H, H
-6c, J = 6.4Hz) ¹³ C-NMR (CDCl ₃ ) δ; 168.013 (C = 0), 109.4
98 (= C ( CH ₃ ) ₂ ), 101.470 (C-1b), 97.850 (C-1a), 97.469
(C-1c), 56.510 (C-2a), 28.011, 26.277 (= C (CH ₃ ) ₂ ),
16.849 (C-6c)

[Reference Example 4] (Synthesis of Compounds [7] and [8]) 2.62 g (2.11 mM) of Compound [6] was mixed with trifluoroacetic acid / THF / water (20 ml / 25 ml / 5 ml) mixed solvent. After dissolution, the mixture was stirred under ice cooling for 4 hours and at room temperature for 2 hours. The reaction solution was extracted with ethyl acetate, washed successively with water, saturated aqueous sodium hydrogen carbonate and saturated brine, and dried over magnesium sulfate.
The solvent was distilled off. The residue was purified by column chromatography (toluene / ethyl acetate = 3/1 development) to give 517 mg of compound [7] (yield, 31.3%) and compound [8] .1.
72 g (yield, 68.0%) was obtained. Compound [7] Rf = 0.08 (toluene / ethyl acetate = 3/1) [α] _D ²⁷ + 6.3 ° (C = 0.73, CHCl ₃ ) Elemental analysis value: C ₄₄ H ₄₇ NO ₁₂ = 781.866 Calculated value as C; 67.59 H; 6.06 N; 1.79 Measured value C; 67.60 H; 6.10 N; 1.63 ¹ H-NMR (400 MHz, CDCl ₃ ) δ; 7.85-7.
16 (m, 19H, aromatic H), 5.739 (m, 1H, -CH ₂ - CH = CH
₂ ), 5.256 (d, 1H, H-1a, J = 8.5Hz), 5.132 (dd, 1H, =
CH ₂ ,, J = 1.5, 17.4Hz), 5.047 (dd, 1H, = CH ₂ ,, J = 1.5, 1
0.4Hz), 4.340 (d, 1H, H-1b, J = 7.3Hz), 4.289 (ddt,
1H, O -CH ₂ -CH = CH ₂ , J = 1.2, 4.9, 13.1Hz), 4.056 (ddt,
1H, O- CH ₂ CH = CH ₂ , J = 1.2, 6.4, 13.1Hz) ¹³ C-NMR (CDCl ₃ ) δ; 103.537 (C-1b), 97.3
61 (C-1a), 56.239 (C-2a)

Compound [8] Rf = 0.45 (toluene / ethyl acetate = 3/1) [α] _D ²⁷ -11.4 ° (C = 0.76, CHCl ₃ ) Elemental analysis value: C ₇₁ H ₇₅ Calculated value with NO ₁₆ = 1198.386 C; 71.16 H; 6.31 N; 1.17 Measured value C; 71.70 H; 6.38 N; 1.49 ¹ H-NMR (400 MHz, CDCl ₃ ) δ; 7.72-7.
02 (m, 34H, aromatic H), 5.652 (m, 1H, -CH ₂ - CH = CH
₂ ), 5.162 (d, 1H, H-1a, J = 8.5Hz), 5.074 (dd, 1H, =
CH ₂ ,, J = 1.8, 17.4Hz), 4.986 (dd, 1H, = CH ₂ ,, J = 1.5, 1
0.4Hz), 1.086 (d, 3H, H-6c, J = 6.4Hz) ¹³ C-NMR (CDCl ₃ ) δ; 168.013 (C = 0), 101.6
95 (C-1b), 97.742 (C-1a), 97.469 (C-1c), 56.294 (C
-2a), 16.795 (C-6c)

Reference Example 5 (Compound

Synthesis of [9]] 450 mg (0.37 mM) of compound [8] was dissolved in acetic anhydride / pyridine / 4-dimethylaminopyridine 12 ml / 2 ml / catalyst amount), and the mixture was stirred overnight at room temperature. The reaction solution was extracted with ethyl acetate, washed successively with water, saturated sodium bicarbonate water, diluted hydrochloric acid, and saturated saline, and dried over magnesium sulfate,
The solvent was distilled off. The residue was purified by column chromatography (toluene / ethyl acetate = 6/1 development) to obtain 420 mg of an oily substance (yield, 87.3%). Rf = 0.66 (toluene / ethyl acetate = 5/1) [α] _D ²⁶ −7.9 ° (C = 0.80, CHCl ₃ ) Elemental analysis value: Calculated as C ₇₅ H ₇₉ NO ₁₈ = 1282.462 C; 70.24 H; 6.21 N; 1.09 Measured value C; 70.27 H; 6.22 N; 0.98 ¹ H-NMR (400 MHz, CDCl ₃ ) δ; 7.72-6.
99 (m, 34H, aromatic H), 5.639 (m, 1H, -CH ₂ - CH = CH
₂ ), 5.328 (d, 1H H-4b, J = 2.7Hz), 5.109 (d, 1H, H-1
a, J = 8.5Hz), 5.065 (dd, 1H, = CH ₂ , J = 1.5, 17.4Hz),
4.983 (dd, 1H, = CH ₂ , J = 1.5, 10.4Hz), 4.826 (d, 1H,
H-1c, J = 3.6Hz), 4.566 (d, 1H, H-1b, J = 7.6Hz), 3.884
(dd, 1H, H-3c, J = 2.7, 10.1Hz), 3.777 (dd, 1H, H-2
c, J = 3.6, 10.1Hz), 3.519 (d, 1H, H-4c, J = 3.1Hz),
1.911, 1.749 (2 × S, 6H, CO CH ₃ ), 1.128 (d, 3H, H-6
C, J = 6.4 Hz), ¹³ C-NMR (CDCl ₃ ) δ; 169.747, 169.530, 16
8.013 (C = 0), 101.968 (C-1b), 97.794 (C-1a), 97.469
(C-1c), 56.402 (C-2a), 20.588 (CO CH ₃ ), 17.012 (C-
6c)

Reference Example 6 (Synthesis of Compound [10]) Compound

[9] 405 mg (0.316 mM), palladium chloride 265 mg (1.5 mM) and sodium acetate 123 mg
(1.5 mM) was dissolved in 90% acetic acid and stirred at room temperature for 4 hours. The reaction solution was diluted with ethyl acetate, the insoluble matter was filtered off from Celite, the filtrate was washed successively with water, saturated aqueous sodium bicarbonate solution and saturated brine, dried over magnesium sulfate, and the solvent was evaporated. The residue was purified by column chromatography (toluene / ethyl acetate = 5/1 development) to give an oil 2
78 mg (yield, 70.9%) were obtained. Rf = 0.58 (toluene / ethyl acetate = 2/1) [α] _D ²⁷ + 1.6 ° (C = 0.84, CHCl ₃ ) Elemental analysis value: C ₇₂ H ₇₅ NO ₁₈ = 1242.396 Calculated value C 69.61 H; 6.08 N; 1.13 Measured value C; 69.40 H; 6.09 N; 1.04 ¹ H-NMR (400 MHz, CDCl ₃ ) δ; 7.71-6.
89 (m, 34H, Aromatic H), 5.324 (d, 1H, H-4b, J =
2.7Hz), 4.830 (d, 1H, H-1c, J = 3.6Hz), 3.877 (dd, 1
H, H-3c, J = 2.7, 10.1Hz), 3.784 (dd, 1H, H-2c, J = 3.
6, 10.4Hz), 1.917, 1.745 (2 × S, 6H, COCH ₃ ), 1.135
(d, 3H, H-6c, J = 6.4Hz)

Reference Example 7 (Synthesis of Compound [11]) 232 mg of Compound [10] (0.1) under an argon gas atmosphere.
86 mM) and 288 mg of trichloroacetonitrile (2
(mM) was dissolved in 5 ml of dichloromethane, and 30 μl (0.2 mM) of DBU was added thereto under ice cooling and stirred for 1 hour. The reaction solution was directly purified by column chromatography (toluene / ethyl acetate = 5/1 development) to give oil 2
08 mg (yield, 81.2%) was obtained. Rf = 0.46 (toluene / ethyl acetate = 5/1) [α] _D ²⁷ + 6.5 ° (C = 1.0, CHCl ₃ ) ¹ H-NMR (400 MHz, CDCl ₃ ) δ; 8.528
(s, 1H, NH), 7.69-6.99 (m, 34H, aromatic H), 6.394
(d, 1H, H-1a, J = 8.8Hz), 5.328 (d, 1H, H-4b, J = 2.9
Hz), 4.828 (d, 1H, H-1c, J = 3.6Hz), 4.627 (d, 1H, H
-1b, J = 8.0Hz), 3.885 (d, d, 1H, H-3c, J = 2.4, 10.2H
z), 3.786 (dd, 1H, H-2c, J = 3.6, 10.2Hz), 1.915, 1.
759 (2 × S, 6H, COCH ₃ ), 1.300 (d, 3H, H-6c, J = 6.6H
z)

[Reference Example 8] (Synthesis of Compounds [13] and [14]) MS (AW) sufficiently dried in advance under an argon gas atmosphere.
400) Compound [1
1] 578 mg (0.42 mM) and [12] 605 mg (0.4
62 mM) was dissolved in 15 ml of dichloroethane and added. −
BF ₃ · Et ₂ O 50μ under 23 ℃ (CCl ₄ -Dry ice)
1 (0.42 mM) was added and stirred for 1 hour. Triethylamine was added, and after the reaction was completed, it was diluted with chloroform,
The insoluble matter was filtered off from Celite, and the filtrate was washed successively with water, saturated sodium bicarbonate water and saturated saline, dried over magnesium sulfate, and then the solvent was distilled off. The residue was purified by column chromatography (n-hexane / ethyl acetate = 5/1 development) to obtain 472 mg of compound [13] (yield, 51.1%) and 90 mg of compound [14] (yield, 17.5%). %), And 372 mg of the compound [12] (yield, 61.5%) was recovered.

The compound [13] Rf = 0.40 (n-hexane / ethyl acetate = 2/1) [α] ^{_{D 27 -23.5 ° (C = 0.52}} , CHCl 3) Elemental analysis: C ₁₃₃ H ₁₃₇ NO ₂₈ = 2197.564 Calculated value C; 72.69 H; 6.28 N; 0.64 Measured value C; 72.22 H; 6.24 N; 0.70 ¹ H-NMR (400 MHz, CDCl ₃ ) δ; 7.38-6.
84 (m, 69H, aromatic H), 5.351 (d, 1H, H-4d, J =
3.6Hz), 5.345 (d, 1H, H-1c, J = 8.2Hz), 1.926,1.740
(2 × S, 6H, CO CH ₃ ), 1.131 (d, 3H, H-6e, J = 6.7 Hz) ¹³ C-NMR (CDCl ₃ ) δ; 169.695, 169.478, 16
7.632 (C = 0), 102.454 (C-1a, C-1b), 102.020 (C-1d),
99.961 (C-1c), 97.577 (C-1e), 57.106 (C-2c), 20.5
34 (CO CH ₃ ), 16.958 (C-6e)

Compound [14] Rf = 0.29 (n-hexane / ethyl acetate) [α] _D ²⁴ -41.4 ° (C = 0.69, CHCl ₃ ) Elemental analysis value: C ₇₂ H ₇₄ NO ₁₇ = 1225.389 calculated value C; 70.57 H; 6.09 N; 1.14 measured value C; 70.63 H; 6.03 N; 1.08 ¹ H-NMR (400 MHz, CDCl ₃ ) δ; 7.72-7.
04 (m, 34H, Aromatic H), 6.714 (s, 1H, H-1a), 5.
439 (d, 1H, H-4b, J = 2.7Hz), 4.955 (dd, 1H, H-3b, J =
3.6, 10.4Hz), 4.797 (d, 1H, H-1c, J = 3.4Hz), 3.380
(d, 1H, H-4c, J = 1.5Hz), 2.039, 1.952 (2 × S, 6H, CO
CH ₃ ), 0.836 (d, 3H, H-6c, J = 6.7Hz) ¹³ C-NMR (C
DCl ₃ ) δ; 170.015, 169.851, 168.013 (C = 0), 10
7.655 (C-1a), 102.779 (C-1b), 98.661 (C-1c), 20.642
(CO CH ₃ ), 16.525 (C-6c)

[Reference Example 9] (Synthesis of compound [15]) 446 mg (0.203 mM) of compound [13] was dissolved in methanol / THF (10 ml / 4 ml), and 0.2 N N was added thereto.
2 ml of aOMe methanol solution was added, and the mixture was stirred at room temperature for 2 hours. After completion of the reaction, Amberlyst 15 was added to neutralize the reaction solution, which was then filtered off, and the filtrate was evaporated. The residue was purified by column chromatography (toluene / ethyl acetate = 5/1 development) to obtain 374 mg of an oily substance (yield, 87.2%). Rf = 0.46 (toluene / ethyl acetate = 3/1) [α] ^{_{D 25 -21.6 ° (C = 0.69}} , CHCl 3) Elemental analysis: Calculated C ₁₂₉ H ₁₃₃ NO ₂₆ = 2113.488 C; 73.31 H; 6.34 N; 0.66 measured value C; 73.42 H; 6.38 N; 0.70 ¹ H-NMR (400 MHz, CDCl ₃ ) δ; 7.52-7.
00 (m, 69H, aromatic H), 5.400 (d, 1H, H-1c, J = 8.
5Hz), 1.087 (d, 3H, H-6e, J = 6.4Hz) ¹³ C-NMR (CDCl ₃ ) δ; 167.683 (C = 0), 102.
454 (C-1a, C-1b), 101.804 (C-1d), 99.961 (C-1c), 9
7.742 (C-1e), 57.052 (C-2c), 16.904 (C-6e),

Example 1 (Synthesis of Compounds [17] and [18]) MS (AW) which was sufficiently dried in advance under an argon gas atmosphere.
400) 500 mg in a 2 diameter brown flask containing compound [15] 211 mg (0.1 mM) and [16] 182 mg
(0.14 mM) was dissolved in 7 ml of dichloroethane and added.
-23 ° C (carbon tetrachloride / dry ice), BF ₃ · Et ₂ O
12 μl (0.1 mM) was added and stirred for 1 hour. Triethylamine was added and after the reaction was completed, the reaction mixture was diluted with chloroform, the insoluble matter was removed by filtration from Celite, the filtrate was washed successively with saturated aqueous sodium bicarbonate solution and saturated brine, dried over magnesium sulfate, and the solvent was distilled off. did. The residue was subjected to gel filtration (Bio-boads S
-X2, Nippon Bio-Rad Laboratories, developed with benzene) and column chromatography (developed with toluene / ethyl acetate = 4/1), compound [17] 252 mg
(Yield, 77.8%), Compound [18] 20.0 mg (Yield,
12.0%) and 14 mg of compound [15] (yield, 6.6%)
Was recovered.

Compound [17] Rf = 0.72 (toluene / ethyl acetate) [α] _D ²⁶ 27.4 ° (C = 0.92, CHCl ₃ ) Elemental analysis value: C ₁₉₁ H ₁₉₈ N ₂ O _45. Calculated value with H ₂ O = 3259.707 C; 70.38 H; 6.18 N; 0.86 Measured value C; 70.07 H; 6.09 N; 0.96 ¹ H-NMR (400 MHz, CDCl ₃ ) δ; 7.41-6.
79 (m, 93H, aromatic H), 5.316 (d, 1H, H-1f, J = 8.
3Hz), 5.236 (d, 1H, H-4h, J = 2.7Hz), 5.148 (d, 1H, H-1c,
J = 8.3Hz), 5.008 (dd, 1H, H-2h, J = 8.3,10.3Hz), 1.999,
1.982, 1.947, 1.824 (4 × S, 12H, COCH ₃ ), 1.194 (d, 3
H, H-6g, J = 6.4Hz), 0.999 (d, 3H, H-6e, J = 6.4Hz) ¹³ C-NMR (CDCl ₃ ) δ; 169.851, 169,634, 16
8,446, 167,849 (C = 0), 102.402, 101.643, 99.961, 99.
584, 98.934, 97.469 (C-1a-h), 57.106, 56.510 (C-2
c, f), 21.347, 20.371 (CO CH ₃ ), 16.687 (C-6e, g)

The compound [18] Rf = 0.40 (toluene / ethyl acetate) [α] ^{_{D 24 -36.9 ° (C = 0.51}} , CHCl 3) Elemental _{analysis: C 62 H 65 NO 19:} 1 / 2H ₂ O = 1137.212 calculated C; 65.48 H; 5.85 N; 1.23 measurements C; 65.10 H; 5.79 N; 1.23 1 H-NMR (400MHz, CDCl 3) δ; 7.73-7.
04 (m, 24H, aromatic H), 6.686 (s, 1H, H-1a), 5.
372 (d, 1H, H-4b, J = 2.4Hz), 5.259 (dd, 1H, H-2b, J = 8.2,1
0.7Hz), 4.984 (dd, 1H, H-3b, J = 3.3,10.7Hz), 4.824 (d,
1H, H-1c, J = 3.0Hz), 4.713, (d, 1H, H-1b, J = 7.9Hz), 3.747
(q, 1H, H-5c, J = 6.1Hz), 3.390 (d, 1H, H-4c, J = 1.5Hz), 2.
163, 2.064, 1.988, 1.924, (4 × S, 12H, COCH ₃ ), 0.8
43 (d, 3H, H-6c, J = 6.4Hz)

Example 2 (Synthesis of Compound [19]) 252 mg (0.077 mM) of compound [17] was dissolved in 20 ml of 2% hydrazine-ethanol, and the mixture was refluxed and stirred overnight.
The solvent was evaporated, the residue was dissolved in acetic anhydride / pyridine / 4-dimethylaminopyridine (2 ml / 2 ml / catalytic amount), and the mixture was stirred at room temperature for 6 hours. After completion of the reaction, the reaction solution was extracted with ethyl acetate, washed successively with water, saturated sodium bicarbonate water, dilute hydrochloric acid and saturated brine, dried over magnesium sulfate, and the solvent was evaporated. Column chromatography of the residue (ethyl acetate /
Purified with toluene = 1/1 development), 208 mg oil
(Yield, 86.3%) was obtained. Rf = 0.43 (ethyl acetate / toluene = 1/1) [α] ^{_{D 25 -30.9 ° (C = 1.1}} , CHCl 3) Elemental _{analysis: C 181 H 198 N 2 O} 44 · H 2 Calculated value as O = 3123.567 C; 69.60 H; 6.45 N; 0.89 Measured value C; 69.27 H; 6.43 N; 0.84 ¹ H-NMR (400 MHz, CDCl ₃ ) δ; 7.42-7.
06 (m, 85H, aromatic H), 5.501 (d, 1H, NH), 5.44
7 (d, 1H, H-4d, J = 3.3Hz), 5.419 (d, 1H, NH), 5.278 (d, 1H,
H-4h, J = 3.0Hz), 2.019, 1.947, 1.934, 1.862, 1.832,
1.469, 1.266, (7 × S, 21H, COCH ₃ ), 1.170 (d, 3H, HH-
6 e or g , J = 6.4 Hz), 1.114 (d, 3H8, H-6 g or e
, J = 6.4 Hz) ¹³ C-NMR (CDCl ₃ ) δ; 169.747, 169.366, 16
8.828 (C = 0), 102.454, 101.102, 99.745, 97.469, 96.
819 (C-1a-h), 59.111, 56.889 (C-2c, f), 22.972, 2
1.401, 20.859, 20.534 (C OC H ₃ ), 17.121, 16.741 (C-6
e, g)

[Example 3] (Synthesis of compound [20]) 203 mg (0.065) of compound [19] was prepared under a hydrogen gas atmosphere.
mM) and 10% Pd-C 50 mg methanol acetic acid (3 ml
The solution was stirred overnight at room temperature. After completion of the reaction, the catalyst was filtered off, the solvent was distilled off, the residue was dissolved in acetic anhydride / pyridine / 4-DMAP (2 ml / 2 ml / catalyst amount), and the mixture was stirred at room temperature overnight. The reaction solution is extracted with ethyl acetate,
The extract was washed successively with water, saturated sodium bicarbonate water, diluted hydrochloric acid, and saturated brine, dried over magnesium sulfate, and the solvent was evaporated. Column chromatography of the residue (acetone / toluene = 1
1/1 development), 117 mg of oily matter (yield, 78.0
%) Was obtained. Rf = 0.43 (acetone / toluene = 2/1) Elemental analysis value: C ₉₆ H ₁₃₂ N ₂ O ₆₁ = 2290.098 Calculated value C; 50.35 H; 5.81 N; 1.22 Measured value C; 49.98 H; 5.75 N; 1.28 ¹ H-NMR (400 MHz, CDCl ₃ ) δ; 6.254
(d, 1H, H-1aα, J = 3.7Hz), 2.191-1.966 (mS, 72H, COCH
₃ ), 1.203, 1.156, (2 × d, 2 × 3H, H-6e, gJ = 6.4H
z)

[Example 4] (Synthesis of compound [21]) 103 mg (0.045 mM) of compound [20] was added to 2 ml of DMF.
Hydrazine acetate 9.2mg (0.1m
M) was added and the mixture was stirred at room temperature for 1 hour. The reaction solution was extracted with ethyl acetate, washed with water, dried over magnesium sulfate,
The solvent was distilled off. Dissolve the residue in 1 ml of dichloroethane,
To this was added 72 mg (0.5 mM) of trichloroacetonitrile. Argon gas atmosphere, under ice cooling, DBU 7
μl (0.045 mM) was injected and stirred for 1 hour. The reaction solution was directly purified by column chromatography (acetone / toluene = 1/1 development), and the compound [21] 78 mg
(Yield, 72.5%) was obtained. Rf = 0.50 (acetone / toluene = 3/2) [α] ^{_{D 25 -25.0 ° (C = 1.00}} , CHCl 3) 1 H-NMR (400MHz, CDCl 3) δ; 8.651
(s, 1H, = NH), 6.482 (d, 1H, H-1a, J = 3.7Hz), 2.189-
1.901 (ms, 69H, COCH ₃ ), 1.200, 1.153, (2 × d, 2 × 3
(H, H-6e, g, J = 6.6Hz)

[Reference Example 10] (Synthesis of Compounds [23] and [24]) MS (AW) thoroughly dried in advance under an argon gas atmosphere.
300) 300 mg in a 2 diameter brown flask containing compounds [21] 24.0 mg (0.1 mM) and [22] 9.0 mg (0.1 mM).
(01 mM) was dissolved in 2 ml of chloroform and added. -23
° C. (CCl ₄ - dry ice) _{under, BF 3 · Et 2 O 2μl} (0.
(015 mM) was injected and stirred for 1 hour. Triethylamine was added, and after the reaction was completed, the reaction mixture was diluted with chloroform, the insoluble matter was filtered off from Celite, the filtrate was washed successively with saturated aqueous sodium bicarbonate solution and saturated brine, dried over magnesium sulfate, and the solvent was distilled off. did. The residue was subjected to column chromatography (toluene / acetone = 3/1 development) and gel filtration method (Bio-boa
Purified by ds S-X2, Nippon Bio-Rad Laboratories, developed with benzene), and compound [23] 1.5 mg (yield, 4.8)
%) And [24] 6.5 mg (yield 27.2%).

The compound [23] Rf = 0.65 (toluene / acetone = 1/1) [α] ^{_{D 24 -29.3 ° (C = 0.058}} , CHCl 3) 1 H-NMR (400MHz, CDCl 3) δ ; 7.67-7.
30 (m, 10H, aromatic H), 2.188-1.900 (ms, 69H,
COCH ₃ ), 1.202 (d, 3H, H-6 e or g , J = 6.7Hz), 1.15
5 (d, 3H, H-6 g or e , J = 6.4Hz), 1.004 (s, 9H,-
tBu), 0.881 (2 × t, 6H, 2 × CH ₃ (Cer), J = 5.5Hz) Compound [24] Rf = 0.36 (acetone / toluene = 1/1) [α] _D ²⁵ -47. 0 ° (C = 0.67, CHCl ₃ ) Elemental analysis value: Calculated value as C ₉₆ H ₁₃₄ N ₂ O ₆₀・ 2H ₂ O = 2312.146 C; 49.87 H; 6.01 N; 1.21 Measured value C; 49.55 H; 5.79 N; 1.23 ¹ H-NMR (400 MHz, CDCl ₃ ) δ; 2.192-
1.902 (ms, 69H, COCH ₃ ), 1.203, 1.154, (2 × d, 2
× 3H, H-6e, g, J = 6.7Hz)

Reference Example 11 (Synthesis of Compound [1] (2Le ^x )) Compound [23] 1.2 mg (3.8 × 10 ⁻⁴ mM) was added with THF 0.2.
It was dissolved in 5 ml, 5 μl of a THF 1M solution of tetra-n-butylammonium fluoride was added thereto, and the mixture was stirred at room temperature overnight. The reaction solution was distilled off under reduced pressure, and the residue was dissolved in methanol / THF (0.5 ml / 0.5 ml), to which 0.2
Add 0.1 ml of N NaOMe / methanol solution, and add 4 at room temperature.
Stirred for hours. After the reaction was completed, Amberlyst 15 was added to neutralize the reaction solution, and the insoluble solution was filtered off from Celite. After concentrating the filtrate, the residue was purified by gel filtration (LH-20, chloroform / methanol / water = 60/40 / 4.6), and compound [1] 0.5 mg (yield, 4.8% ) Got. Rf = 0.43 (n-butanol / ethanol / water = 2
/ 1/1) ¹ H-NMR (400 MHz, d ⁶ -DMSO) δ; 5.544 (d
f, 1H, H-5Cer, J = 6.1,14.6Hz), 5.357 (dd, 1H, H-4Ce
r, J = 7.3, 15.1Hz), 4.853 (d, 2H, H-1d, g, J = 3.4H
z), 4.730 (d, 1H, H-1c, J = 7.6Hz), 4.714 (d, 1H, H-
1f), 4.624 (q, 2H, H-5d, g, J = 6.1Hz), 4.337 (d, 1
H, H-1e, J = 6.8Hz), 4.291 (d, 1H, H-1h, J = 6.8Hz), 4.
267 (d, 1H, H-1b), 4.168 (d, 1H, H-1a, J = 7.8Hz),
3.065 (t, 1H, H-2a, J = 7.8Hz), 2.026 (t, 1H, H-2′C
er, J = 7.5Hz), 1.907 (s, 6H, 2 × NAc), 1.007 (d, 6
H, H-6d, g, J = 6.3Hz), 0.853 (t, 6H, 2 × CH ₃ Cer,
(J = 6.3Hz)

Claims (5)

[Claims] 1. A glycosphingolipid-related compound represented by the general formula [I]. Embedded image In the formula, R ₁ , R ₂ and R ₃ are independently a hydrogen atom, a benzyl group or an acetyl group, R ₄ is a phthaloylamino group or an acetylamino group, R ₅ is a benzyl group,
An acetyl group or a -C (= NH) -CCl ₃ group. 2. The method according to claim 1, wherein R ₁ is a benzyl group, R ₂ is an acetyl group, R ₃ is a hydrogen atom, R ₄ is a phthaloylamino group, and R ₅ is a benzyl group. Compound of. 3. The method according to claim 1, wherein R ₁ is a benzyl group, R ₂ is an acetyl group, R ₃ is an acetyl group, R ₄ is an acetylamino group, and R ₅ is a benzyl group. Compound. 4. The compound according to claim ₁ , wherein R ₁ , R ₂ and R ₃ are acetyl groups, R ₄ is an acetylamino group and R ₅ is an acetyl group. 5. R ₁ , R ₂ and R ₃ are acetyl groups, R ₄ is an acetylamino group, and R ₅ is -C (= NH) -CC.
The compound according to claim 1, which is an l ₃ group.