JPH0925235A - Suppressant for cancer metastasis containing 1-o-acylglycerol 2,3-phosphate derivative as active ingredient - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new suppressant for cancer metastasis, containing a glycerophospholipid as an active ingredient and having a property of suppressing infiltrations by cancer cells. SOLUTION: This suppressant for cancer metastasis contains a 1-O- acylglycerol 2,3-phosphate derivative of the formula (R is a 2-30C alkyl, alkenyl or alkynyl; any of which may contain a cycloalkane ring ; M is H or a counter cation) as an active ingredient. The derivative can suitably be blended with other usually used ingredients and prepared as a powder, a granule, a capsule, a tablet, a syrup, a suspension, an injectable solution, etc. The daily dose is 1-50mg administered in several divided portions. 1-O-[(Z)-9-Hexadecenoyl(-2,3-0- isopropylidene-sn-glycerol is exemplified as the compound of the formula.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a cancer metastasis inhibitor containing glycerophospholipid as an active ingredient, which is expected to be used as a carcinostatic agent.

[0002]

2. Description of the Related Art Among deaths due to adult diseases, the percentage of those who die due to cancer tends to increase year by year. Although researches on cancer treatment methods and metastasis mechanisms have made rapid progress in recent years, no epoch-making treatment method has been found yet, and treatment with chemotherapeutic agents does not produce sufficient effect. Has not been done. Most cancer deaths are caused by metastases, and suppressing metastases is considered a good measure to reduce cancer deaths. Conventionally, ovostatin, matristatin, aprotinin and the like have been known as cancer metastasis suppressors, but their effects have been insufficient.

On the other hand, a novel lysophosphatidic acid, 1-
O-[(9'S, 10'R) -9 ', 10'-methanohexadecanoyl] -sn-glycerol-2,3-phosphate (PHYLPA) is known to have DNA polymerase α inhibitory activity [K. Mur
akami-Murofushi et al., J. Biol. Chem., 267, 21512-
21517 (1992).]. Also, 1-O-alkanoylglycerol-2,3-phosphate, 1-O-alkenoylglycerol-
2,3-phosphate or 1-O-alkinoylglycerol-2,3-phosphate is known to have a cytostatic effect [Murobushi et al., Bioindustry, 11,
484-496 (1994).]. EP-A-317,968 also describes 1-O-oleylglycerol-2,3-phosphate as an electrostatic liquid developer. However, it has not been reported so far that these compounds have a cancer metastasis inhibitory action.

[0004]

Cancer metastasis begins with the detachment of cancer cells from the primary lesion, destruction of extracellular matrix, invasion into blood vessels, adhesion in distant organs, vascular endothelial cell layer It takes a multi-step process of infiltration and proliferation beyond.
Various factors are involved in these processes, and their inhibitors are drawing attention as new cancer metastasis inhibitors. Among them, invasion is the most characteristic step of the phenomenon of metastasis, and it is considered highly possible that an excellent anticancer agent will be developed from substances that suppress the invasion of cancer cells. An object of the present invention is to provide a novel cancer metastasis inhibitor that can suppress invasion of cancer cells.

[0005]

[Means for Solving the Problems] As a result of intensive investigations by the present inventors on substances that suppress the invasion of cancer cells, a specific 1-O-acylglycerol derivative strongly suppresses the invasion of cancer cells. As a result, they have completed the present invention.

That is, the present invention has the following general formula:

[0007]

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(In the formula, R represents a linear or branched alkyl group, alkenyl group or alkynyl group having 2 to 30 carbon atoms, and the alkyl group, alkenyl group or alkynyl group contains a cycloalkane ring. M represents a hydrogen atom or a counter cation group) 1-O-
The present invention relates to a cancer metastasis inhibitor containing an acylglycerol-2,3-phosphate derivative as an active ingredient.

[0009]

BEST MODE FOR CARRYING OUT THE INVENTION The substituent R in the above formula has 2 to 8 carbon atoms.
Thirty linear or branched alkyl, alkenyl or alkynyl groups, which may contain cycloalkane rings. Specific examples of these substituents R, butyl group, hexyl group, octyl group, undecyl group, pentadecyl group, hexadecyl group, alkyl groups such as heptadecyl group, 4-pentenyl group, 6-heptenyl group,
8-decenyl group, 8-undecenyl group, 8-dodecenyl group, 8-
Pentadecenyl group, 8-heptadecenyl group, heptadeca-
8,11-dienyl group, heptadeca-8,11,14-trienyl group,
Nonadeca-4,7,10,13-tetraenyl group, Nonadeca-4,7,1
Alkenyl groups such as 0,13,16-pentaenyl group and henicosa-3,6,9,12,15,18-hexaenyl group, or alkynyl groups such as 8-decynyl group, 8-pentadecynyl group, 8-heptadecynyl group Can be mentioned. Examples of the cycloalkane ring include a cyclopropane ring, a cyclobutane ring, a cyclopentane ring, a cyclohexane ring, a cyclooctane ring and the like.

When M in the above formula is a counter cation group, examples thereof include sodium ion, potassium ion, lithium ion and ammonium ion.

1-O-acylglycerol according to the present invention
2,3-phosphate can be prepared by the method described in the literature (S. Kobayashi et.
al., Tetrahedron Letters, 34, 4047 (1993).).

The 1-O-acylglycerol-2,3-phosphate of the present invention can be administered orally or parenterally. The dosage form includes, for example, powders, granules,
Capsules such as tablets, pills, syrups, suspensions and injections can be exemplified. These are conventional excipients (for example, starch, lactose, crystalline cellulose, magnesium aluminometasilicate, silicic acid anhydride, mannitol, etc.), binders (for example hydroxypropyl) depending on the patient's symptoms, age, and therapeutic purpose. Cellulose, polyvinylpyrrolidone etc.), lubricants (eg magnesium stearate, talc etc.), disintegrants (eg carboxymethyl cellulose, carboxymethyl cellulose calcium etc.) coating agents (eg hydroxyethyl cellulose) flavoring agents, solubilizers or solubilizing agents (eg Distilled water for injection, physiological saline propylene glycol, etc.), suspension agents (eg surfactants such as polysorbate 80) pH adjusters (eg organic acids or metal salts thereof), aqueous or oily solubilizers (eg alcohol) , Fatty acid esters, etc.), viscosity Manufactured using an ordinary manufacturing method (eg, the method prescribed in the 12th revised Japanese Pharmacopoeia) using a binder (eg, carboxyvinyl polymer, polysaccharides, etc.), an emulsifier (eg, surfactant, etc.), a stabilizer, etc. can do. Furthermore, it is also possible to prepare a sustained-release preparation by a known technique. The dose varies depending on the condition of the patient, age, weight, purpose of treatment and the like, but is generally 1 mg to 50 mg, and is administered once or divided into several times a day.

Hereinafter, the present invention will be described in more detail with reference to production examples, examples and test examples, but it goes without saying that the present invention is not limited thereto.

[0014]

【Example】

 Production Example 1

[0015]

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Metallic lithium (393 mg, 56.6 mmol) was added to liquid ammonia (10 mL) under an argon atmosphere at −78 ° C., and the mixture was stirred until a uniform blue solution was obtained. A tetrahydrofuran solution (3 mL) of 1-O-benzyl-2,3-O-isopropylidene-sn-glycerol (1.05 g, 4.72 mmol) was added to this solution, and the mixture was refluxed for 1 hour. Ammonium chloride (3.03 g, 56.6 mmol) was added in portions and the ammonia was evaporated. Brine was added to the reaction solution, and the product was extracted with ether. The ether layer was washed with saturated saline,
Dry over anhydrous sodium sulfate. Concentrate under reduced pressure to give crude 2,3-
O-isopropylidene-sn-glycerol was obtained. A solution of crude 2,3-O-isopropylidene-sn-glycerol, (Z) -9-hexadecenoic acid (1.00 g, 3.94 mmol) and dimethylaminopyridine (96.1 mg, 0.79 mmol) thus obtained in methylene chloride. A methylene chloride solution (2 mL) of dicyclohexylcarbodiimide (852 mg, 4.13 mmol) was added to (6 mL) at 0 ° C under an argon atmosphere, and the mixture was further stirred at room temperature for 10 hours. The insoluble matter was filtered off, and the insoluble matter was washed with methylene chloride. The methylene chloride layers were combined, washed successively with 2% hydrochloric acid, saturated aqueous sodium hydrogen carbonate solution and saturated brine, and dried over anhydrous sodium sulfate. After concentration under reduced pressure, the residue was purified by silica gel column chromatography (hexane / ethyl acetate = 95/5) and 1-O-[(Z) -9-hexadecenoyl]-
2,3-O-isopropylidene-sn-glycerol (876 mg, 2.
38 mmol, 61%) was obtained as a colorless oil.

¹ H-NMR (400 MHz, CDCl ₃ ): δ = 0.88 (3H,
t, J = 6.8 Hz), 1.24-1.36 (16H, m), 1.37 (3H, s),
1.44 (3H, s), 1.58-1.68 (2H, m), 1.50-2.06 (4H.
m), 2.34 (2H, t, J = 7.5 Hz), 3.74 (1H, dd, J = 6.
2, 8.5 Hz), 4.08 (1H, dd, J = 6.5, 8.5 Hz), 4.09 (1
H, dd, J = 6.0, 11.5 Hz), 4.17 (1H, dd, J = 4.7, 1
1.5 Hz), 4.31 (1H, dddd, J = 4.7, 6.0, 6.2, 6.5 H
z), 5.30-5.40 (2H, m). ¹³ C-NMR (CDCl ₃ ): δ = 14.0, 22.6, 24.8, 25.4, 26.6,
27.1, 27.2, 28.9, 29.0 (2), 29.1, 29.6, 29.7, 31.7,
34.1, 64.5, 66.3, 73.6, 109.8, 129.7, 130.0, 173.
6. IR (neat): 2920, 2850, 1740, 1450, 1370, 1210, 116
0, 1080, 1050 cm ^-1 .EI-MS: m / z = 368 (M ⁺ ), 353 (M ⁺ -Me), 310. HRMS: m / z = 368.2918 (368.2924 calcd for C ₂₂ H ₄₀ O ₄ ,
M ⁺ ). [Α] _D ²⁰ +0.79 (c 1.01, CHCl ₃ ).

Production Example 2

[0019]

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The 1-O-[(Z) -9-hexadecenoyl] -2,3-O-isopropylidene-sn-glycerol (866 mg, 2.35 mmol) obtained in Preparation Example 1 in isopropyl alcohol (10
(mL) -water (2 mL) mixed solution was added pyridinium p-toluenesulfonate (118 mg, 0.47 mmol) and 2.5 hours 80
Heated to ° C. The residue obtained by concentration under reduced pressure was subjected to silica gel chromatography (hexane / ethyl acetate = 7 /
Purification was performed in 3) to obtain 1-O-[(Z) -9-hexadecenoyl] -sn-glycerol (662 mg, 2.02 mmol, yield 86%) as a colorless oily substance.

¹ H-NMR (400 MHz, CDCl ₃ ): δ = 0.89 (3H,
t, J = 6.9 Hz), 1.22-1.38 (16H, m), 1.58-1.68 (2H,
m), 1.97-2.07 (4H, m), 2.35 (2H, t, J = 7.5 Hz),
2.47 (1H, br), 2.86 (1H, br), 3.56-3.64 (1H, brd
d), 3.66-3.74 (1H, brdd), 3.89-3.97 (1H, br), 4.15
(1H, dd, J = 6.0, 11.6 Hz), 4.20 (1H, dd, J = 4.
8,11.6 Hz), 5.29-5.39 (2H, m). ¹ H-NMR (400MHz, CDCl ₃ / D ₂ O): δ = 0.89 (3H, t, J =
6.9 Hz), 1.22-1.38 (16H, m), 1.58-1.68 (2H, m), 1.
97-2.07 (4H, m), 2.35 (2H, t, J = 7.5 Hz), 3.58 (1
H, dd, J = 5.9, 11.6 Hz), 3.68 (1H, dd, J = 3.8, 1
1.6 Hz), 3.92 (1H, dddd, J = 3.8, 4.8, 5.9, 6.0 H
z), 4.15 (1H, dd, J = 6.0, 11.6 Hz), 4.20 (1H, dd,
J = 4.8, 11.6 Hz), 5.29-5.39 (2H, m). ¹³ C-NMR (CDCl ₃ ): δ = 14.1, 22.6, 24.9, 27.1, 27.2,
29.0, 29.06 (2), 29.11, 29.6, 29.7, 31.7, 34.1, 6
3.3, 65.1, 70.2, 129.7, 130.0, 174.3. IR (neat): 3400, 2920, 2850, 1730, 1460, 1180, 112
0, 1050 cm ^-1 .EI-MS: m / z = 328 (M ⁺ ), 310 (M ⁺ -H ₂ O), 297 (M ⁺ -CH ₂ O)
H) .CI-MS: m / z = 329 (M + H ⁺ ), 311 (M ⁺ -OH) .HRMS: m / z = 328.2617 (328.2612 calcd for C ₁₉ H ₃₆ O ₄ ,
M ⁺ ) [α] _D ²⁰ -0.53 (c 1.12, CHCl ₃ ).

Production Example 3

[0023]

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Triazole (118 m
g, 1.71 mmol) in tetrahydrofuran (3 mL), phosphorus oxychloride (53.2 μL, 0.57 mmol) and triethylamine (0.37 mL, 2.67 mmol) were added at 0 ° C., and
The mixture was stirred for 5 minutes to prepare phosphoryl tristriazolide. 1-O-obtained in Production Example 2 was added to the above reaction solution at 0 ° C.
[(Z) -9-Hexadecenoyl] -sn-glycerol (156 mg,
0.476 mmol) in tetrahydrofuran (3 mL)
Stir for 15 minutes. Ether was added to the reaction solution and cooled with ice.
It was poured into 2% hydrochloric acid (50 mL) and extracted with ether. 0.05N aqueous sodium hydroxide was added until the pH of the aqueous layer reached 7. The aqueous layer was freeze-dried to give 1-O-[(Z) -9-hexadecenoyl] -sn-glycerol sodium salt of 2,3-phosphate (1,196 mg, 0.476 mmol, quantitative yield) as a white powder. Obtained.

¹ H-NMR (400 MHz, CD ₃ OD): δ = 0.90 (3H,
t, J = 6.9 Hz), 1.26-1.40 (16H, m), 1.57-1.67 (2H,
m), 2.00-2.08 (8H, m), 2.36 (2H, t, J = 7.4 Hz),
3.94 (1H, ddd, J = 7.0, 9.1, 9.1 Hz), 4.18 (1H, d
d, J = 6.0, 11.8 Hz), 4.23 (1H, dd, J = 5.2, 11.8)
Hz), 4.24 (1H, ddd, J = 6.3, 9.1, 12.7 Hz), 4.52-
4.60 (1H, m), 5.30-5.40 (2H, m). ¹³ C-NMR (CDCl ₃ / CD ₃ OD = 3/1): δ = 13.5, 22.2, 24.4, 2
6.7, 26.8, 28.6, 28.7 (2), 28.8, 29.29, 29.32, 31.
4, 33.6, 63.8 (d, J _CP = 6.2 Hz), 65.5, 72.9 (d, J
_CP = 2.5 Hz), 129.3, 129.6, 173.7. IR (KBr): 2920, 2850, 1730, 1460, 1250, 1140, 1090
cm ^-1 .SI-MS: m / z = 435 (M + Na ⁺ ), 413 (M + H ⁺ ).

Production Example 4

[0027]

[Chemical 6]

In the same manner as in Production Example 1, liquid ammonia (10 mL), metallic lithium (311 mg, 44.8 mmol), 1-O-
Benzyl-2,3-O-isopropylidene-sn-glycerol (8
28 mg, 3.73 mmol) was used to prepare 2,3-O-isopropylidene-sn-glycerol. The crude 2,3-O-isopropylidene-sn-glycerol thus obtained was treated in the same manner as in Production Example 1 with (E) -9-hexadecenoic acid (527 mg, 2.07 mmol) and dicyclohexylcarbodiimide (470 mg, 2.28 mmo
l), dimethylaminopyridine (50.6 mg, 0.42 mmol)
1-O-[(E) -9-hexadecenoyl] -2,3-O-isopropylidene-sn-glycerol (707 mg, 1.92 mmol, 9
3%) as a colorless oil.

[0029]¹H-NMR (400MHz, CDCl_Three): δ = 0.88 (3H,
t, J = 6.9 Hz), 1.24-1.36 (16H, m), 1.37 (3H, s),
1.44 (3H, s), 1.58-1.68 (2H, m), 1.92-2.02 (4H,
m), 2.34 (2H, t, J = 7.5 Hz), 3.74 (1H, dd, J = 6.
2, 8.5 Hz), 4.08 (1H, dd, J = 6.5, 8.5 Hz), 4.09 (1
H, dd, J = 6.0, 11.5 Hz), 4.17 (1H, dd, J = 4.5, 1
1.5 Hz), 4.31 (1H, dddd, J = 4.5, 6.0, 6.2, 6.5 H
z), 5.32-5.44 (2H, m).¹³ C-NMR (CDCl_Three): δ = 14.1, 22.6, 24.9, 25.4, 26.7,
28.8, 28.9, 29.05, 29.27, 29.5, 29.6, 31.7, 32.5,
32.6, 34.0, 64.5, 66.3, 73.7, 109.8, 130.2,130.5,
173.6. IR (neat): 2920, 2850, 1740, 1450, 1370, 1250, 121
0, 1160, 1080, 1050 cm ^-1.EI-MS: m / z = 368 (M⁺), 353 (M⁺-Me), 310.HRMS: m / z = 368.2948 (368.2925 calcd for C_{twenty two}H₄₀O_Four,
 M⁺). [α]_D ²⁰ +2.3 (c 1.04, CHCl_Three).

Production Example 5

[0031]

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1-O-[(E) -9-hexadecenoyl] -2,3-O-isopropylidene-sn-glycerol (696 mg, 1.89 mmol) obtained in Preparation Example 4 and pyridinium p-toluenesulfonate. (95.1 mg, 0.38 mmol) was reacted in the same manner as in Production Example 2 to give 1-O-[(E) -9-hexadecenoyl] -sn-glycerol (557 mg, 1.70 mmol, yield 90%) as a colorless oil. Obtained as a substance.

[0033]¹H-NMR (400MHz, CDCl_Three): δ = 0.88 (3H,
t, J = 6.9 Hz), 1.22-1.39 (16H, m), 1.58-1.68 (2H,
 m), 1.91-2.02 (4H, m), 2.35 (2H, t, J = 7.5 Hz),
2.54 (1H, brdd, J = 6.0, 6.3 Hz) D_TwoO exchangeable,
 2.91 (1H, brd, J = 5.0 Hz) D_TwoO exchangeable, 3.60
(1H, ddd, J = 6.0, 6.0, 11.6 Hz), 3.70 (1H, ddd, J
= 3.8, 6.3, 11.6 Hz), 3.93 (1H, ddddd, J = 3.8, 4.
9, 5.0, 6.0, 6.0 Hz), 4.14 (1H, dd, J = 6.0, 11.6
Hz), 4.19 (1H, dd, J = 4.9, 11.6 Hz), 5.32-5.44 (2
H, m).¹³ C-NMR (CDCl_Three): δ = 14.1, 22.6, 24.9, 28.8, 28.9,
 29.1 (2), 29.5, 29.6, 31.7, 32.5, 32.6, 34.1, 63.
3, 65.1, 70.2, 130.2, 130.5, 174.3. IR (neat): 3350, 2920, 2850, 1730, 1460, 1390, 125
0, 1210, 1180, 1060 cm ^-1.EI-MS: m / z = 328 (M⁺), 310 (M⁺-H_TwoO), 297 (M⁺-CH_TwoO
H). CI-MS: m / z = 329 (M + H⁺), 311 (M⁺-OH) .HRMS: m / z = 328.2636 (328.2612 calcd for C₁₉H₃₆O_Four,
 M⁺). Anal. Calcd for C₁₉H₃₆O_Four: C, 69.47; H, 11.05. Foun
d: C, 69.34; H, 11.19. [α]_D ²⁰ -0.56 (c 1.06, CHCl_Three).

Production Example 6

[0035]

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In the same manner as in Production Example 3, triazole (116 mg, 1.68 mmol) in tetrahydrofuran (3 mL),
Phosphoryl tris triazolide was prepared from phosphorus oxychloride (52.0 μL, 0.56 mmol) and triethylamine (0.36 mL, 2.61 mmol). The phosphoryl tristriazolide thus prepared and 1-O-[(E) prepared in Preparation Example 5
-9-Hexadecenoyl] -sn-glycerol (153 mg, 0.46
6 mmol) was reacted in the same manner as in Production Example 3 to obtain 1-O-[(E) -9
The sodium salt of -hexadecenoyl] -sn-glycerol 2,3-phosphate (2,192 mg, 0.466 mmol, quantitative yield) was obtained as a white powder.

¹ H-NMR (400 MHz, CD ₃ OD): δ = 0.90 (3H,
t, J = 6.9 Hz), 1.25-1.40 (16H, m), 1.56-1.66 (2H,
m), 1.93-2.03 (4H, m), 2.36 (2H, t, J = 7.4 Hz),
3.94 (1H, ddd, J = 7.0, 9.1, 9.1 Hz), 4.18 (1H, d
d, J = 6.1, 11.8 Hz), 4.23 (1H, dd, J = 5.2, 11.8
Hz), 4.24 (1H, ddd, J = 6.3, 9.1, 12.7 Hz), 4.51-
4.60 (1H, m), 5.33-5.43 (2H, m). ¹³ C-NMR (CDCl ₃ / CD ₃ OD = 3/1): δ = 13.5, 22.2, 24.4, 2
8.4, 28.5, 28.67, 28.69, 29.1, 29.2, 31.3, 32.10,
32.15, 33.5, 63.8 (J _CP = 6.2 Hz), 65.5, 72.9 (J
_CP = 2.4 Hz), 129.8, 130.1, 173.7.IR (KBr): 2920, 2850, 1740, 1460, 1250, 1180, 1140
cm ^-1 .SI-MS: m / z = 435 (M + Na ⁺ ), 413 (M + H ⁺ ).

Production Example 7

[0039]

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In the same manner as in Production Example 1, liquid ammonia (15 mL), metallic lithium (484 mg, 69.7 mmol), 1-O-
Benzyl-2,3-O-isopropylidene-sn-glycerol (6
2,3-O-isopropylidene-sn-glycerol was prepared from 46 mg, 2.91 mmol). The crude 2,3-O-isopropylidene-sn-glycerol thus obtained was treated in the same manner as in Production Example 1 with (Z) -9-octadecenoic acid (1.07 g, 3.78 mmol) and dicyclohexylcarbodiimide (719 mg, 3.49 mmo
l), dimethylaminopyridine (70.9 mg, 0.58 mmol)
1-O-[(Z) -9-octadecenoyl] -2,3-O-isopropylidene-sn-glycerol (594 mg, 1.50 mmol, 5
2%) as a colorless oil.

[0041]¹H-NMR (400MHz, CDCl_Three): δ = 0.88 (3H,
t, J = 6.8 Hz), 1.23-1.36 (21H, m), 1.37 (3H, s),
1.44 (3H, s), 1.58-1.68 (2H, m), 1.97-2.40 (4H,
m), 2.34 (1H, t, J = 7.5 Hz), 3.74 (1H, dd, J = 6.
2, 8.5 Hz), 4.08 (1H, dd, J = 6.5, 8.5 Hz), 4.09 (1
H, dd, J = 6.0, 11.5 Hz), 4.17 (1H, dd, J = 4.7, 1
1.5 Hz), 4.32 (1H, dddd, J = 4.7, 6.0, 6.2, 6.5 H
z), 5.29-5.40 (2H, m).¹³ C-NMR (CDCl_Three): δ = 14.1, 22.6, 24.9, 25.3, 26.7,
 27.1, 27.2, 29.1 (3), 29.3 (2), 29.5, 29.7 (2), 31.9,
 34.1, 64.5, 66.3, 73.7, 109.8, 129.7, 130.0, 173.
6. IR (neat): 2920, 2850, 1740, 1460, 1370, 1250, 121
0, 1160, 1090, 1060 cm ^-1.EI-MS: m / z = 381 (M⁺-Me), 338.CI-MS: m / z = 381 (M⁺-Me), 339.HRMS: m / z = 381.3014 (381.3003 calcd for C_{twenty three}H₄₁O_Four,
 M⁺-Me). [Α]_D ²⁰ +1.55 (c 1.03, CHCl_Three).

Production Example 8

[0043]

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1-O-[(Z) -9-octadecenoyl] -2,3-O-isopropylidene-sn-glycerol (479 mg, 1.21 mmol) obtained in Preparation Example 8 and pyridinium p-toluenesulfonate (60.8 mg, 0.24 mmol) was reacted in the same manner as in Production Example 2 to give 1-O-[(Z) -9-octadecenoyl] -sn-glycerol (387 mg, 1.09 mmol, yield 90%) as a colorless oil. Obtained as a substance.

¹ H-NMR (400 MHz, CDCl ₃ ): δ = 0.88 (3H,
t, J = 6.8 Hz), 1.22-1.38 (20H, m), 1.59-1.60 (2H,
m), 1.96-2.06 (4H, m), 2.35 (2H, t, J = 7.5 Hz),
2.47 (1H, d, J = 4.9 Hz) D ₂ O exchangeable, 3.60 (1
H, ddd, J = 5.7, 5.7, 11.6 Hz), 3.70 (1H, ddd, J =
4.1, 6.3, 11.6 Hz), 3.94 (1H, ddddd, J = 4.1, 4.
8, 4.9, 5.7, 6.1 Hz), 4.15 (1H, dd, J = 6.1, 11.7
Hz), 4.22 (1H, dd, J = 4.8, 11.7 Hz), 5.30-5.40 (2
H, m). ¹³ C-NMR (CDCl ₃ ): δ = 14.1, 22.7, 24.9, 27.2 (2), 2
9.1 (2), 29.2, 29.3 (2), 29.5, 29.7, 29.8, 31.9, 3
4.2, 63.4, 65.2, 70.3 129.7, 130.1, 174.4.IR (neat): 3400, 2920, 2850, 1740, 1460, 1380, 118
0, 1120, 1050 cm ^-1 .EI-MS: m / z = 356 (M ⁺ ), 338 (M ⁺ -H ₂ O), 325 (M ⁺ -CH ₂ O
H) .CI-MS: m / z = 357 (M + H ⁺ ), 339 M ⁺ -HO).

Production Example 9

[0047]

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In the same manner as in Preparation Example 3, triazole (65.8 mg, 0.953 mmo) was added to tetrahydrofuran (2 mL).
l), phosphorus oxychloride (46.2 μL, 0.50 mmol), and triethylamine (0.32 mL, 2.31 mmol) were used to prepare phosphoryl tristriazolide. The phosphoryl tristriazolide thus prepared and 1-O- prepared in Preparation Example 8
[(Z) -9-Octadecenoyl] -sn-glycerol (103 mg,
1.49 mmol) was reacted in the same manner as in Production Example 3 to obtain 1-O-
[(Z) -9-Octadecenoyl] -sn-glycerol sodium salt of 2,3-phosphate (3, 162 mg, 0.368 mmol, yield 8
9%) as a white powder.

¹ H-NMR (400 MHz, CD ₃ OD): δ = 0.91 (3H,
t, J = 6.9 Hz), 1.27-1.38 (20H, m), 1.56-1.67 (2H,
m), 1.99-2.09 (4H, m), 2.37 (2H, t, J = 7.4 Hz),
3.94 (1H, ddd, J = 7.0, 9.1, 9.1 Hz), 4.19 (1H, d
d, J = 6.0, 11.8 Hz), 4.23 (1H, dd, J = 5.0, 11.8
Hz), 4.24 (1H, dd, J = 6.3, 9.1, 12.7 Hz), 4.52-4.
60 (1H, m), 5.30-5.40 (2H, m). ¹³ C-NMR (CDCl ₃ / CD ₃ OD = 3/1): δ = 13.7, 22.3, 24.5, 2
6.8, 26.9, 28.8 (2), 28.9, 29.0 (2), 29.2, 29.4
(2), 31.6, 33.6, 63.9 (d, J _CP = 6.3Hz), 65.6, 73.0
(d, J _CP = 2.2 Hz), 129.4, 129.7, 173.7. IR (KBr): 2920, 2850, 1730, 1630, 1460, 1240, 118
0, 1140, 1090 cm ^-1 .SI-MS: m / z = 463 (M + Na ⁺ ), 441 (M + H ⁺ ).

Production Example 10

[0051]

Embedded image

In the same manner as in Production Example 1, liquid ammonia (10 mL), metallic lithium (396 mg, 57.1 mmol), 1-O-
Benzyl-2,3-O-isopropylidene-sn-glycerol (1.06g, 4.76 mmol) to 2,3-O-isopropylidene-sn-
Glycerol was prepared. The crude 2,3-O-obtained in this way
Using isopropylidene-sn-glycerol in the same manner as in Production Example 1, 9-hexadecynoic acid (1.00 g, 3.97 mmol), dicyclohexylcarbodiimide (858 mg, 4.17 mmol),
Dimethylaminopyridine (96.8 mg, 0.79 mmol) was reacted to give 1-O- (9-hexadecinoyl) -2,3-O-isopropylidene-sn-glycerol (759 mg, 2.07 mmol, 52%) as a colorless oily substance. Obtained.

¹ H-NMR (400 MHz, CDCl ₃ ): δ = 0.89 (3H,
t, J = 6.9 Hz), 1.20-1.42 (12H, m), 1.37 (3H, s),
1.44 (3H, s), 1.43-1.52 (4H, m), 1.63 (2H, quinte
t, J = 7.6Hz), 2.11-2.17 (4H, m), 2.34 (2H, t, J =
7.6 Hz), 3.74 (1H, dd, J = 6.2, 8.5 Hz), 4.08 (1H,
dd, J = 6.5, 8.5 Hz), 4.09 (1H, dd, J = 6.1, 11.5
Hz), 4.17 (1H, dd, J = 4.7, 11.5 Hz), 4.32 (1H, dd
dd, J = 4.7, 6.1, 6.2, 6.5 Hz). ¹³ C-NMR (CDCl ₃ ): δ = 14.0, 18.67, 18.71, 22.5, 24.
8, 25.4, 26.6, 28.5, 28.6, 28.7, 28.97, 29.02, 29.
1, 31.3, 34.0, 64.5, 66.3, 73.6, 80.0, 80.3, 109.7,
173.5. IR (neat): 2920, 2850, 1740, 1460, 1370, 1210, 116
0, 1080, 1050 cm ^-1 .EI-MS: m / z = 366 (M ⁺ ), 351 (M ⁺ -Me). HRMS: m / z = 366.2769 (376.2768 calcd for C ₂₂ H ₃₈ O ₄ ,
M ⁺ ). [Α] _D ²⁰ +0.20 (c 0.96, CHCl ₃ ).

Production Example 11

[0055]

Embedded image

1-O- (9-hexadecinoyl) -2,3-O-isopropylidene-sn-glycerol obtained in Preparation Example 10 (74
9 mg, 2.05 mmol) and pyridinium p-toluenesulfonate (103 mg, 0.41 mmol) were reacted in the same manner as in Production Example 2 to obtain 1-O- (9-hexadecinoyl) -sn-glycerol (599 mg, 1.84 mmol). , Yield 90%) was obtained as a colorless oily substance.

¹ H-NMR (400 MHz, CDCl ₃ ): δ = 0.89 (3H,
t, J = 7.0 Hz), 1.22-1.42 (12H, m), 1.42-1.52 (4H,
m), 1.58-1.68 (2H, m), 2.10-2.18 (4H, m), 2.35 (2
H, t, J = 7.6 Hz), 2.56 (1H, br, D ₂ O exchangeable),
2.93 (1H, br, D ₂ O exchangeable), 3.60 (1H, dd, J
= 5.9, 11.5 Hz), 3.70 (1H, dd, J = 3.9, 11.5 Hz),
3.93 (1H, dddd, J = 3.9, 4.9, 5.9, 6.0 Hz), 4.15
(1H, dd, J = 6.0, 11.6Hz), 4.19 (1H, dd, J = 4.9,
11.6 Hz). ¹³ C-NMR (CDCl ₃ ): δ = 14.0, 18.66, 18.70, 22.5, 24.
8, 28.5, 28.6, 28.7, 28.95, 29.00, 29.1, 31.3, 34.
1, 63.3, 65.1, 70.2, 80.0, 80.3, 174.3.IR (neat): 3400, 2920, 2850, 1730, 1460, 1410, 138
0, 1330, 1280, 1250, 1210, 1180, 1120, 1090, 1050,
990, 930 cm ^-1 .EI-MS: m / z = 326 (M ⁺ ), 295 (M ⁺ -CH ₂ OH) .CI-MS: m / z = 327 (M + H ⁺ ), 309 (M ⁺ -OH) .HRMS: m / z = 326.2443 (326.2454 calcd for C ₁₉ H ₃₄ O ₄ ,
M ⁺ ). [Α] _D ²⁰ -0.70 (c 1.14, CHCl ₃ ).

Production Example 12

[0059]

Embedded image

In the same manner as in Production Example 3, triazole (107 mg, 1.56 mmol) in tetrahydrofuran (2 mL),
Phosphoryl tris triazolide was prepared from phosphorus oxychloride (48.4 μL, 0.52 mmol) and triethylamine (0.34 mL, 2.4 mmol). The phosphoryl tristriazolide thus prepared and 1-O- (9 prepared in Preparation Example 11
-Hexadecinoyl) -sn-glycerol (141 mg, 0.433
mmol) in the same manner as in Production Example 3 to give 1-O- (9-hexadecinoyl) -sn-glycerol 2,3-phosphate sodium salt (4, 171 mg, 0.417 mmol, yield 95%) in white. Obtained as a powder.

¹ H-NMR (400 MHz, CD ₃ OD): δ = 0.91 (3H,
t, J = 7.0 Hz), 1.24-1.50 (16H, m), 1.58-1.68 (2H,
m), 2.09-2.16 (4H, m), 2.37 (2H, t, J = 7.5 Hz),
3.94 (1H, ddd, J = 7.0, 9.1, 9.1 Hz), 4.18 (1H, d
d, J = 5.7, 11.8 Hz), 4.23 (1H, dd, J = 5.2, 11.8
Hz), 4.25 (1H, ddd, J = 6.3, 9.1, 12.7 Hz), 4.52-
4.60 (1H, m). ¹³ C-NMR (CDCl ₃ / CD ₃ OD = 3/1): δ = 13.5, 18.28, 18.32,
22.2, 24.4, 28.1, 28.3, 28.4, 28.67, 28.70, 28.7
5, 31.0, 33.6, 63.8 (d, J _CP = 6.2 Hz), 65.5, 73.0
(d, J _CP = 6.2 Hz), 79.7, 80.0, 173.6. IR (KBr): 2930, 2850, 1730, 1240, 1180, 1140, 1100
cm ^-1 .SI-MS: m / z = 433 (M + Na ⁺ ), 411 (M + H ⁺ ).

Production Example 13

[0063]

Embedded image

In the same manner as in Production Example 1, liquid ammonia (30 mL), metallic lithium (382 mg, 55.1 mmol), 1-O-
Benzyl-2,3-O-isopropylidene-sn-glycerol (1.02g, 4.59 mmol) to 2,3-O-isopropylidene-sn-
Glycerol was prepared. The crude 2,3-O-obtained in this way
Isopropylidene-sn-glycerol was reacted with palmitic acid (1.29 g, 5.05 mmol), dicyclohexylcarbodiimide (993 mg, 4.82 mmol) and dimethylaminopyridine (112 mg, 0.92 mmol) in the same manner as in Production Example 1
-O-palmitoyl-2,3-O-isopropylidene-sn-glycerol (1.17 g, 3.17 mmol, 69%) was obtained as a colorless oily substance.

¹ H-NMR (400 MHz, CDCl ₃ ): δ = 0.88 (3H,
t), 1.22-1.34 (24H, m), 1.37 (3H, s), 1.44 (3H, s),
1.58-1.68 (2H, m), 2.34 (2H, t), 3.74 (1H, dd, J
= 6.2, 8.5 Hz), 4.08 (1H, dd, J = 6.5, 8.5 Hz), 4.
09 (1H, dd, J = 5.9, 11.5 Hz), 4.17 (1H, dd, J =
4.7, 11.5 Hz), 4.32 (1H, dddd, J = 4.7, 5.9, 6.2,
6.5 Hz). ¹³ C-NMR (CDCl ₃ ): δ = 14.1, 22.7, 24.9, 25.4, 26.7,
29.1, 29.2, 29.3, 29.4, 29.6 (2), 29.7 (4), 31.9,
34.1, 64.5, 66.3, 73.7, 109.8, 173.6. IR (KBr): 2920, 2850, 1730, 1470, 1390, 1370, 124
0, 1220, 1190, 1170, 1160, 1080, 1050, 1030, 850 c
m ^-1 .EI-MS: m / z = 355 (M ⁺ -CH ₃ ). Elemental analysis (C ₂₂ H ₄₂ O ₄ ): Calculated value; C 71.31, H 11.42: Measured value; C 71.30, H 11.53. [α] _D ²⁰ +0.55 (c 1.08, CHCl ₃ ).

Production Example 14

[0067]

Embedded image

1-O-palmitoyl-obtained in Production Example 13
2,3-O-isopropylidene-sn-glycerol (243 mg, 0.
657 mmol) and pyridinium p-toluenesulfonate (3
3.0 mg, 0.13 mmol) was reacted in the same manner as in Production Example 2 to obtain 1-O-palmitoyl-sn-glycerol (212 mg, 0.642).
mmol, yield 98%) was obtained as a colorless oily substance.

¹ H-NMR (400 MHz, CDCl ₃ ): δ = 0.88 (3H,
t), 1.19-1.37 (24H, m), 1.58-1.68 (2H, m), 2.05 (1
H, t, J = 6.3 Hz), 2.35 (2H, t, J = 7.5 Hz), 2.50
(1H, d, J = 5.2 Hz), 3.60 (1H, ddd, J = 5.7, 6.3,
11.5 Hz), 3.70 (1H, ddd, J = 4.0, 6.3, 11.5 Hz), 3.
94 (1H, ddddd, J = 4.0, 4.7, 5.2, 5.7, 6.1 Hz), 4.
15 (1H, dd, J = 6.1, 11.7 Hz), 4.21 (1H, dd, J =
4.7, 11.7 Hz). ¹³ C-NMR (CDCl ₃ ): δ = 14.1, 22.6, 24.9, 29.1, 29.2,
29.3, 29.4, 29.6 (5), 31.7, 34.1, 63.3, 65.1, 70.
2, 174.4.IR (KBr): 3350, 2900, 2850, 1730, 1460, 1390, 122
0, 1190, 1180, 1100, 1050 cm ^-1 .EI-MS: m / z = 299 (M ⁺ -H). Elemental analysis (C ₁₉ H ₃₈ O ₄ ): Calculated value; C 69.05, H 11.59: Measured Value; C 68.87, H 11.71. [Α] _D ²⁰ +0.84 (c 0.95, CHCl ₃ ) .mp 67-69 ℃.

Production Example 15

[0071]

Embedded image

In the same manner as in Production Example 3, triazole (65.8 mg, 0.95 mmo) was added to tetrahydrofuran (2 mL).
l), phosphorus oxychloride (29.6 μL, 0.32 mmol), and triethylamine (0.21 mL, 1.5 mmol) were used to prepare phosphoryl tristriazolide. The phosphoryl tristriazolide thus prepared and 1-prepared in Preparation Example 14
O-palmitoyl-sn-glycerol (87.4 mg, 0.27 mmo
l) was reacted in the same manner as in Production Example 3 to obtain the sodium salt of 1-O-palmitoyl-sn-glycerol 2,3-phosphate (5, 88.3 mg, 0.213 mmol, yield 81%) as a white powder. .

¹ H-NMR (400 MHz, CD ₃ OD): δ = 0.90 (3H,
t, J = 6.9 Hz), 1.27-1.35 (24H, m), 1.56-1.66 (2H,
m), 2.36 (2H, t, J = 7.0, 9.1 Hz), 4.16-4.29 (3H,
m), 4.52-4.60 (1H, m) .IR (KBr): 2900, 2850, 1730, 1470, 1240, 1140, 800
cm ^-1 .

Production Example 16

[0075]

Embedded image

In the same manner as in Production Example 1, liquid ammonia (15 mL), metallic lithium (232 mg, 33.4 mmol), 1-O-
Benzyl-2,3-O-isopropylidene-sn-glycerol (6
2,3-O-isopropylidene-sn-glycerol was prepared from 18 mg, 2.78 mmol). The crude 2,3-O-isopropylidene-sn-glycerol thus obtained was treated in the same manner as in Production Example 1 with eicosanoic acid (1.04 g, 3.34 mmol), dicyclohexylcarbodiimide (630 mg, 3.06 mmol) and dimethylamino. React with pyridine (67.8 mg, 0.56 mmol) 1-
O-eicosanoyl-2,3-O-isopropylidene-sn-glycerol (756 mg, 1.77 mmol, 64%) was obtained as white crystals.

¹ H-NMR (400 MHz, CDCl ₃ ): δ = 0.88 (3H,
t, J = 6.9 Hz), 1.20-1.37 (32H, m), 1.37 (3H, s),
1.44 (3H, s), 1.57-1.68 (2H, m), 2.34 (2H, t, J =
7.6 Hz), 3.74 (1H, dd, J = 6.2, 8.5 Hz), 4.08 (1H,
dd, J = 6.5, 8.5 Hz), 4.09 (1H, dd, J = 5.9, 11.5
Hz), 4.17 (1H, dd, J = 4.7, 11.5 Hz), 4.32 (1H, dd
dd, J = 4.7, 5.9, 6.2, 6.5 Hz). ¹³ C-NMR (CDCl ₃ ): δ = 13.8, 22.4, 24.6, 25.1, 26.4,
28.8, 28.9, 29.1, 29.2, 29.3, 29.4 (9), 31.6, 33.
8, 64.2, 66.1, 73.4, 109.5, 173.4. IR (KBr): 2910, 2850, 1740, 1470, 1380, 1240, 121
0, 1170, 1080, 1050, 840 cm ^-1 .EI-MS: m / z = 411 (M ⁺ -CH ₃ ). Elemental analysis (C ₂₆ H ₅₀ O ₄ ): Calculated value; C 73.19, H 11.81: Found; C 72.93, H 11.72. [Α] _D ²⁰ +1.50 (c 0.93, CHCl ₃ ) .mp 48-51 ° C.

Production Example 17

[0079]

Embedded image

1-O-eicosanoyl obtained in Production Example 16
-2,3-O-isopropylidene-sn-glycerol (354 mg,
0.831 mmol) and pyridinium p-toluenesulfonate (41.8 mg, 0.17 mmol) were reacted in the same manner as in Production Example 2 to obtain 1-O-eicosanoyl-sn-glycerol (321 mg, 0.1.
813 mmol, quantitative yield) was obtained as white crystals.

¹ H-NMR (400 MHz, CDCl ₃ ): δ = 0.88 (3H,
t, J = 6.9 Hz), 1.22-1.35 (32H, m), 1.98-2.08 (1H,
br), 2.35 (2H, t, J = 7.6 Hz), 2.46-2.52 (1H, b
r), 3.56-3.64 (1H, brdd), 3.66-3.75 (1H, brdd), 3.
90-3.98 (1H, br), 4.15 (1H, dd, J = 6.1, 11.7 Hz),
4.22 (1H, dd, J = 4.6, 11.7 Hz). ¹³ C-NMR (CDCl ₃ ): δ = 14.1, 22.7, 24.9, 29.1, 29.2,
29.3, 29.4, 29.6 (3), 29.7 (7), 31.9, 34.1, 63.3,
65.2, 70.3, 174.4.IR (KBr): 3350, 2910, 2850, 1730, 1460, 1390, 119
0, 1180, 1100, 1050 cm ^{- 1} .EI-MS: m / z = 386 (M ⁺ ). Elemental analysis (C ₂₃ H ₄₆ O ₄ ): Calculated value; C 71.45, H 11.99: Measured value; C 71.34 , H 11.82. [Α] _D ²⁰ +2.40 (c 0.83, CHCl ₃ /MeOH=1/1).mp 80-81.5 ° C.

Production Example 18

[0083]

Embedded image

In the same manner as in Production Example 3, triazole (73.4 mg, 1.06 mmo) was added in tetrahydrofuran (2 mL).
l), phosphorus oxychloride (33.0 μL, 0.35 mmol), and triethylamine (0.23 mL, 1.65 mmol) were used to prepare phosphoryl tristriazolide. The phosphoryl tristriazolide thus prepared and 1-prepared in Preparation Example 17
O-Eicosanoyl-sn-glycerol (114 mg, 0.295 mm
ol) was reacted in the same manner as in Production Example 3 to obtain the sodium salt of 1-O-eicosanoyl-sn-glycerol 2,3-phosphate (6, 105 mg, 0.223 mmol, yield 75%) as a white powder. .

¹ H-NMR (400 MHz, CD ₃ OD): δ = 0.90 (3H,
t, J = 6.9 Hz), 1.27-1.33 (32H, m), 1.56-1.66 (2H,
m), 2.36 (2H, t, J = 7.5 Hz), 3.94 (1H, dt, J =
7.1, 9.1 Hz), 4.16-4.29 (3H, m), 4.52-4.61 (1H,
m). IR (KBr): 2910, 2850, 1730, 1470, 1240, 1170, 114
0, 1100, 800 cm ^-1 .

Production Example 19

[0087]

[Chemical 21]

PHYLPA is a method described in the literature (S. Kobayashi e
Tetrahedron Letters, 34, 4047 (1993).).

Test Example 1

Methods described in the literature (H. Akedo et al., Cance
r Res., 46, 2416 (1986) and M. Mukai et al., ibid.,
47, 2167 (1987.), the mesentery of normal rats was adjusted to 0.
Treated with 25% trypsin, transferred single cells (peritoneal mesothelial cells) were transferred to a 35 mm culture dish, and 10% fetal bovine serum was added to Eagle-MEM (2x amino acids, 2x vitamins) medium (MEM). Culture was performed in the presence of liquid to form a monolayer. MEM without culture medium and without fetal bovine serum
Rat ascites hepatoma cells (MM1) (2x10 ⁵ ) were suspended in 2 mL, and 5 mM 1-O-oleoyllysophosphatidic acid (L
PA) (5 μL, final concentration: 12.5 μM) was added to overlay the mesothelial cell layer. 20 hours later, the supernatant (including uninfiltrated cancer cells) was removed, the remaining monolayer was fixed with 10% formalin, and the number of cancer cells invading under the mesothelial cell layer was counted under a microscope. The infiltration ability was calculated by converting the number of cells per 1 cm ² . On the other hand, each 1-O-acylglycerol-2,3-phosphate derivative contained 1% (w / v) bovine serum albumin-phosphate buffer
-Dissolve in physiological saline (PBS) and test compound solution (5 mM)
Was prepared. Test compound solution 5 mM, 5 μL (final concentration: 12.5
μM) or 10 μL (final concentration: 25 μM) 5 mM LPA 5 μL
(Final concentration: 12.5 μM) was added to MM1 suspended in MEM and overlaid on the peritoneal mesothelial cell layer, and the number of invading cancer cells was measured 20 hours later. In Table 1, Production Examples 3, 6, 9, 12, 1
7 examples produced in 5, 18 (1, 2, 3, 4, respectively)
5, 6 and PHYLPA), the number of invading cells when each compound was added to the number of invading cancer cells when the test compound was not added was shown as an inhibition rate.

[0091]

[Equation 1] Inhibition rate = 100− (Number of invading cancer cells when added / Number of invading cancer cells when not added) × 100

[0092]

[Table 1]

[0093]

INDUSTRIAL APPLICABILITY The cancer metastasis inhibitor containing 1-O-acylglycerol 2,3-phosphate of the present invention as an active ingredient has an excellent activity of suppressing invasion of cancer cells, Is expected to be useful.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Rikuko Mukai 4-2-8 Kusunoki-cho, Nada-ku, Hyogo Prefecture

Claims (1)

[Claims] (1) The following general formula: (In the formula, R represents a linear or branched alkyl group having 2 to 30 carbon atoms, an alkenyl group or an alkynyl group, and the alkyl group, alkenyl group or alkynyl group may contain a cycloalkane ring. , M represents a hydrogen atom or a counter cation group), a cancer metastasis inhibitor comprising a 1-O-acylglycerol-2,3-phosphate derivative as an active ingredient.