JPH0617307B2 - Antitumor agent - Google Patents

Description

TECHNICAL FIELD The present invention relates to an antitumor agent.

More specifically, the present invention provides the formula [Wherein R ¹ represents alkyl, R ² , R ³ and R ⁴ each represent hydrogen or lower alkyl, or Is a 5- to 6-membered cyclic ammonio group which may have N, S or O as a constituent atom and which may have a substituent, and A represents a lower alkylene] Alternatively, the present invention provides an antitumor agent containing a salt thereof.

BACKGROUND ART A compound represented by the formula (I) is disclosed in JP-A-55-28955.
Some are included in the scope of claims described in Japanese Patent Publication,
The patent publication only describes specific examples of glycerol derivatives in which the 1-position is acyl (ester bond), and the compound represented by the formula (I) of the present invention [where 1-position is alkyl (ether bond)] Glycerol derivative] is not disclosed at all. The 1-position acyl glycerol derivative disclosed in the patent publication has a 1-position acyl group that is easily hydrolyzed and inactivated in vivo, and thus has a higher activity strength and sustainability than the 1-position alkyl glycerol derivative. Inferior. In fact, lysolecithin does not activate macrophages even at about 1000 times the concentration of PAF, and its antibody-producing ability (PFC) and in vitro and in vivo antitumor activity is higher than that of the corresponding alkyl ether compound, lyso PAF. It is known to be significantly inferior.

On the other hand, the compound of the present invention represented by the formula (I) is less susceptible to such enzymatic degradation and inactivation, and has a long-lasting antitumor effect and is also strong.

It also has the formula as a natural phospholipid compound. [In the formula, R represents hexadecyl or octadecyl]
Platelet activating factor (PAF) represented by is known.

Synthetic phospholipid compounds similar to the compound (II) have PAF-like effects such as platelet activating action, neutrophil activating action, tissue-damaging action, although there are differences in strength based on structural differences.
It is known to have vascular permeability enhancing action, blood pressure lowering action and the like. On the other hand the formula as a derivative of natural phosphatidylcholine A synthetic phospholipid compound represented by the following formula is known (for example, JP-A-52-134027). The compound (III
Unlike natural phospholipid, a) has an antitumor action, but is also known to have a platelet aggregation action [D.
J. Hanahan et al., Biochem. Biophys. Res. C
ommun., 99 , 183 (1981)]. Such effects on platelets may cause cardiovascular disorders such as cerebral thrombosis and angina. In addition, compound (IIIa) has a hypotensive action and a local stimulating action, and all of these actions are side effects, which limits its use as a medicine.

In addition, literature [eg, Thrombosis Res., 30 , 143 (1
983)] is an expression Although the compound represented by the formula (1) is described, the compound has a platelet aggregation action, and its use as a drug is restricted like the compound (IIIa).

Further, in JP-A-57-67589, the formula is Although a synthetic phospholipid represented by the formula [wherein R represents tridecyl or tetradecyl] is described, it has a high toxicity such as a relatively low maximum tolerated dose (LD ₅₀ ) value, and thus it is still problematic for use as a medicine. It is left.

Problems to be Solved by the Invention Synthetic phospholipid compounds have actions such as platelet aggregation action and blood pressure lowering action generally as described above. These effects are side effects when using a synthetic phospholipid compound as an antitumor agent, and since the effective amount capable of exerting an antitumor effect and the side effect expression amount are extremely close,
As it is, it is difficult to use it as an antitumor agent.

The present inventors have conducted intensive studies for the purpose of increasing the drug treatment index, that is, the side effect expression amount / therapeutically effective dose. As a result, the inventors of the present invention have shown that the 2-acetoacetylglycerol compound represented by the formula (I) exhibits a remarkable antitumor activity by intravenous or intraperitoneal administration, and also has a macrophage activating effect. It was found that the effects that were thought to be parallel to the antitumor activity, such as platelet aggregation and blood pressure lowering, were surprisingly significantly weakened, and as a result, the drug treatment index was dramatically improved,
The present invention has been completed.

Means for Solving the Problems [Wherein R ¹ represents alkyl, R ² , R ³ and R ⁴ each represent hydrogen or lower alkyl, or Is a 5- to 6-membered cyclic ammonio group which may have N, S or O as a constituent atom and which may have a substituent, and A represents a lower alkylene] Alternatively, the present invention provides an antitumor agent containing a salt thereof.

Regarding the above formula (I), the alkyl group represented by R ¹ is
For example, n-tetradecyl, n-pentadecyl, n-hexadecyl, n-heptadecyl, n-octadecyl, n
-Eicosanyl, 3,7,11-trimethyldodecyl,
Examples thereof include linear or branched alkyl groups having about 14 to 20 carbon atoms such as 3,7,11,15-tetramethylhexadecyl, and among them, alkyl groups having about 15 to 19 carbon atoms are preferable.

R ² , R ³ and R ⁴ each represent hydrogen or lower alkyl, and examples of the lower alkyl group include C _1-5 alkyl groups (eg, methyl, ethyl, propyl, butyl, pentyl), and preferably It is methyl.

The cyclic ammonio group represented by is a pyridinio group, an oxazolio group, a thiazolio group, a pyridazinio group,
Examples thereof include quinolinio group, isoquinolinio group, pyrrolidinio group, piperidinio group, and these groups further include C
_It may have a substituent such as _1-4 alkyl group (eg, methyl, ethyl, propyl, butyl), hydroxy group, hydroxyethyl group, aminoethyl group, amino (imino) group, carbamoyl group, ureido group, etc. . In the cyclic ammonio group, any ^two groups of R ² , R ³ and R ⁴ form a ring with a quaternary nitrogen atom, and the remaining one group is, for example, C _1-4.
When it is alkyl (eg, methyl, ethyl, propyl, butyl), it is specifically an N-methylpyrrolidinio group, N-
Methylmorpholinio group, N-methylpiperidinio group, N
-A case where a group such as a methylpiperazinio group is formed is included.

Examples of the lower alkylene group represented by A include alkylene groups having about 2 to 5 carbon atoms such as ethylene, trimethylene, tetramethylene and pentamethylene, and among them, trimethylene or tetramethylene is preferable.

In the compound (I), there are two kinds of stereoisomers of R-coordinate and S-coordinate, and each of them or a mixture thereof and a racemate are included in the present invention.

The compound (I) has, for example, the formula [In the formula, X ⁻ represents anion such as chlorine ion, bromide ion, or iodine ion] and [Wherein M ⁺ represents an alkali metal (eg, Na, K) ion or an alkaline earth metal (eg, Ca, Mg) ion], which may be present in the form of a salt, and such a salt may be present. As the above, a pharmacologically acceptable salt is preferable.

The compound (I) of the present invention can be produced, for example, by the following method.

formula [Wherein R ¹ has the same meaning as defined above] was prepared [Helv. Chim. Acta, 65 , 1059 (198
2)], or a synthesis according to a method similar thereto], Compound (IV)
Expression [Wherein A represents the same meaning as described above, X and Y represent halogen (eg, chlorine, bromine, iodine)], and after the reaction, water is reacted to form a compound [Wherein each symbol has the same meaning as defined above] is obtained. The compound (VI) has the formula [Wherein each symbol has the same meaning as defined above], the compound represented by the formula [Wherein each symbol has the same meaning as defined above], and then the compound (VIII) is subjected to a catalytic reduction reaction known per se to give a compound of the formula [Wherein each symbol has the same meaning as defined above] is obtained. Compound (IX) obtained above in an inert solvent,
Compound (I) is obtained by reacting diketene under anhydrous conditions in the presence of a primary amine (eg, pyridine, triethylamine, etc.).

Also, in formula (I), A compound in which is a secondary, tertiary or quaternary amino group has the formula
(I) Medium, Wherein R is a primary, secondary or tertiary amino group and a compound of the formula RI [wherein R represents lower (C ^1-5 ) alkyl] or a compound of the formula (R) ₂ SO ₄ Wherein R represents lower (C _1-5 ) alkyl] or the formula R—O—SO ₂ —R ′ [wherein R represents lower (C _1-5 ) alkyl and R ′ represents Lower alkyl (C _1-4 ) alkyl or p-tolyl] can also be obtained by reacting the compound. The reaction is usually performed in a suitable solvent (eg, acetone, benzene, toluene, dichloromethane, chloroform, tetrahydrofuran) at 0 to 200 ° C.

Compound (IX) is also available in the literature [eg, Helvetica Chemica Act
a, 66 , 1210 (1983)],
It can be obtained by the following method.

formula [Wherein R ¹ is as defined above] is reacted with compound (V), and after the reaction, water is allowed to react to form a compound of the formula [Wherein each symbol has the same meaning as defined above] is obtained. The compound (XI) is reacted with the compound (VII) to give a compound of the formula After obtaining the compound represented by the formula [wherein each symbol has the same meaning as defined above], the compound (XII) is hydrolyzed to obtain the compound (IX). The hydrolysis reaction is preferably performed in the presence of tetraalkylammonium hydroxide (eg, tetra-n-butylammonium hydroxide).

In addition, compound (VIII) can also be produced by the following method.

Compound (IV) with formula After reacting the compound represented by or phosphorus oxychloride, by reacting with water, the formula A compound represented by the formula: wherein R ¹ is as defined above is obtained. The compound (XIV) has the formula [Wherein each symbol has the same meaning as described above, Z ⁻ represents an anion. Is a condensing agent [eg, trichloroacetonitrile, 2,4,6-trimethylbenzenesulfonyl chloride, 2,4,6-triisopropylbenzenesulfonyl chloride, 2,4,6-trimethylbenzenesulfonyl imidazole] , 2,4,6-Triisopropylbenzenesulfonyl-3-nitroazolide, etc.) to give compound (VIII).

In addition, compound (VIII) can also be obtained by reacting compound (IV) with phosphorus oxychloride, then reacting with compound (XV) without allowing water to act, and allowing water to act.

The representative production methods of compound (I) are described above, but the production method of compound (I) used in the present invention is not limited to these methods.

A compound in which A is alkylene having 3 or more carbon atoms, and A is ethylene and R ¹ has 14 to 15 carbon atoms or 1
Compounds that are 7-20 alkyl are new compounds.

Compound (I) can be administered per se or with a pharmacologically acceptable carrier.

Examples of the dosage form of the compound (I) antitumor agent include various pharmaceutical compositions such as injections, tablets, capsules, solutions and ointments, which can be safely administered parenterally or orally.

For the preparation of injections, drip injections, etc., for example, use physiological saline or an aqueous solution containing glucose or other auxiliary agent,
It is performed according to the usual method. Tablets, capsules and the like can also be prepared according to a conventional method. These dosage forms are in the form of dosage units depending on the purpose of administration, for example, in the case of injection, intravenous,
It is used by an appropriate administration route such as subcutaneous administration or direct administration to the affected area.

Action Compound (I) significantly reduces side effects (eg, platelet aggregation, hypotension, vascular permeability enhancement, tissue damage) and enhances main effects (eg, antitumor activity, macrophage activation) It can be administered as a safe antitumor agent to cancer-bearing warm-blooded animals. The administration method, administration route, and dose can be appropriately selected according to the administration subject and symptoms, but the dose for cancer-bearing warm-blooded animals is usually compound (I).
0.1 to 150 mg / Kg (body weight), preferably 2 to
It is about 50 mg / kg body weight. As for the number of administrations, the drug can be applied about once to three times a day, or at intervals of 2 to 7 days. Further, in order to maintain the drug concentration in the tissue at a required level for a long time, intravenous drip infusion may be performed for a long time.

Examples Reference Example 1 3-Hydroxypropyltrimethylammonium Tosylate 1,3-Propanediol 76 g (1.0 mol) was dissolved in 100 ml of triethylamine to give 95 g of tosyl chloride.
(0.50 mol) was added, and the mixture was stirred overnight at room temperature. The reaction mixture was concentrated under reduced pressure, 800 ml of dichloromethane was added to the residue, and water (150 ml), 1N hydrochloric acid (120 ml) and water (15 ml) were added.
The extract was washed with 0 ml) and saturated aqueous sodium hydrogen carbonate (100 ml) in that order, and dried over anhydrous magnesium sulfate. After the desiccant was filtered off, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel (600 g) column, eluting with dichloromethane-methanol (96: 4). The target fraction was concentrated under reduced pressure to obtain 1,3-propanediol monotosylate as a colorless oil. Yield 86.9 g (yield 78%) NMR (90 MHz, CDCl ₃ ) δ: 1.72 to 2.05 (2H, m), 2.
43 (3H, s), 3.69 (2H, t, J = 6Hz), 4.17 (2H, t, J = 6Hz), 7.33
(2H, d, J = 8Hz), 7.79 (2H, d, J = 8Hz) .IR (Neat) cm ^-1 : 3350,2930,2860,1360,1190,1175,965,9
30,815. To 2.0 g (9.2 mmol) of the above tosylate was added 7 ml of a 18% trimethylamine toluene solution, and the mixture was allowed to stand at room temperature for 3 days and the precipitated crystals were collected by filtration. After washing with toluene and drying under reduced pressure,
3-Hydroxypropyltrimethylammonium tosylate was obtained as colorless needles. Melting point 80 ℃ yield 2.3g
(Yield 92%). NMR (90 MHz, CDCl ₃ + CD ₃ OD) δ: 1.80 to 2.
11 (2H, m), 2.37 (3H, s), 3.13 (9H, s), 3.49 ~ 3.73 (2H, m), 3.
92 (2H, m), 7.21 (2H, d, J = 8Hz), 7.75 (2H, d, J = 8Hz) .IR (KBr) cm ^-1 : 3350,1625,1485,1205,1190,1130,1070 , 1
Reference Example 2 5-hydroxypentyltrimethylammonium tosylate 6.2 g of 1,5-pentanediol was prepared in the same manner as in Reference Example 1.
(60 mmol) and tosyl chloride 5.7 g (30 mmol) gave 1,5-pentanediol monotosylate as a colorless oil. Yield 4.1 g (53% yield). NMR (90 MHz, CDCl ₃ ) δ: 1.20 to 1.70 (6H, m), 2.
03 (1H, s), 2.43 (3H, s), 3.53 (2H, t, J = 6Hz), 4.00 (2H, t, J
= 6.5Hz), 7.32 (2H, d, J = 8Hz), 7.76 (2H, d, J = 8Hz) .IR (Neat) cm ^-1 : 3330,2930,1590,1350,1185,1170,950,8
10. 5-Hydroxypentyltrimethylammonium tosylate was obtained as colorless needle crystals from 2.0 g (7.8 mmol) of the above tosylate and trimethylamine. Melting point 143-1
44 ° C. Yield 2.3 g (yield 95%) NMR (90 MHz, d ₆ -DMSO) δ: 1.20 to 1.80 (6H,
m), 2.29 (3H, s), 3.03 (9H, s), 3.17 ~ 3.50 (4H, m), 4.40 (1H,
t, J = 5Hz), 7.11 (3H, t, J = 8Hz), 7.50 (2H, t, J = 8Hz) .IR (KBr) cm ^-1 : 3400,2950,1485,1215,1190,1170,1
115,1027,1005,820,680. Reference Example 3 2- (benzyloxy) -3- (octadecyloxy)
Propyl 3-trimethylammoniopropyl phosphate chloroform 20 ml, phosphorus oxychloride 0.91 g (5.9 mmol) and triethylamine 3.9 m (29 mmol)
A solution of 2-benzyloxy-3-octadecyloxypropanol (2.5 g, 5.8 mmol) in chloroform (40 ml) was added dropwise to the mixture under ice cooling over 30 minutes. Then, the mixture was stirred at room temperature for 1 hour and then ice-cooled to obtain 3-hydroxypropyltrimethylammonium tosylate obtained in Reference Example 1.
A solution of 3 g (8.4 mmol) of pyridine in 80 ml was added dropwise.
The reaction mixture was stirred at room temperature for 3 days, an aqueous solution of 3.9 g of sodium hydrogencarbonate was added to this, and the mixture was concentrated to dryness under reduced pressure. Chloroform-toluene (1: 1) 10 was added to the residue.
0 ml was added, the insoluble material was filtered off, and the filtrate was concentrated under reduced pressure. Then, 70 ml of chloroform was added to the residue, the insoluble material was removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was converted to silica gel (60
g) column chromatography, eluting with chloroform-methanol-water (65: 25: 4),
Concentrate the desired fraction to give 2- (benzyloxy) -3-
(Octadecyloxy) propyl 3-trimethylammoniopropyl phosphate was obtained as a colorless solid. Yield 2.3 g (yield 65%). Thin layer chromatography [silica gel, chloroform-methanol-water (65: 50: 8)] Rf = 0.18 single spot. NMR (90 MHz, CDCl ₃ ) δ: 0.87 (3H, m), 1.24 (30
H, s), 1.45 (2H, m), 1.97 (2H, m), 3.05 (9H, m), 3.26 ~ 4.33 (1
1H, m), 4.63 (2H, s), 7.29 (5H, m) .IR (KBr) cm ^-1 : 3420,2920,2850,1620,1480,1465,1
260,1120,1050,935,840,730. Reference Example 4 2- (benzyloxy) -3- (octadecyloxy)
Propyl 5-trimethylammoniopentyl phosphate 2.5 g (5.8 mmol) of 2-benzyloxy-3-octadecyloxypropanol and 2.3 g of 5-hydroxypentyltrimethylammonium tosylate obtained in Reference Example 2 in the same manner as in Reference Example 3. From (7.6 mmol), 2- (benzyloxy) -3- (octadecyloxy) propyl 5-trimethylammoniopentyl phosphate was obtained as a colorless solid product. Yield 2.8 g (yield 76%). Thin layer chromatography [silica gel, chloroform-methanol-water (65: 50: 8)] Rf = 0.18 single spot NMR (90 MHz, CDCl ₃ ) δ: 0.87 (3H), 1.25 (30H),
1.46 ~ 1.90 (8H), 3.10 (9H), 3.27 ~ 3.53 (4H), 3.70 ~ 3.95
(4H), 4.15 (3H), 4.66 (2H), 7.30 (5H) .IR (KBr) cm ^-1 : 3400,2920,2850,1465,1235,1115,1
100,1067,820. Reference Example 5 2- (Hydroxy) -3- (octadecyloxy) propyl 3-trimethylammoniopropyl phosphate 2.0 g of 2-benzyloxy compound obtained in Reference Example 3 (3.3 mmol) % Water-containing acetic acid 35 ml, and the mixture was stirred in the presence of 10% palladium-carbon 0.5 g in a hydrogen atmosphere at room temperature for 3 hours. The catalyst was filtered off from the reaction mixture, and the filtrate was concentrated under reduced pressure. The residue was chromatographed on a column of silica gel (30g), chloroform-methanol-
Elution with water (65: 25: 4) gave the desired fraction. This was concentrated under reduced pressure to obtain 2- (hydroxy) -3- (octadecyloxy) propyl 3-trimethylammoniopropyl phosphate as a colorless solid product. Yield 1.
4 g (yield 82%). Thin layer chromatography [silica gel, chloroform-methanol-water (65: 50: 8)] Rf = 0.09 single spot NMR (90 MHz, CDCl ₃ + CD ₃ OD) δ: 0.86 (3H,
m), 1.26 (30H, s), 1.53 (2H, m), 2.10 (2H, m), 3.21 (9H, s), 3.
33〜4.06 (11H, m). IR (KBr) cm ^-1 : 3410,2920,2845,1630,1465,1220,1
115,1050,945,850,715,680. Reference Example 6 2- (Hydroxy) -3- (octadecyloxy) propyl 5-trimethylammoniopentyl phosphate In the same manner as in Reference Example 5, the 2-benzyloxy form obtained in Reference Example 4 The target product was obtained as a colorless solid product from 2.5 g (3.9 mmol). Yield 1.8 g (84% yield) Thin layer chromatography [silica gel, chloroform-methanol-water (65: 50: 8)] Rf = 0.09 single spot NMR (90 MHz, CDCl ₃ ) δ: 0.87 (3H), 1.26 (30H),
1.40 ~ 2.03 (8H), 3.23 (9H), 3.30 ~ 3.56 (5H), 3.70 ~ 3.97
(4H), 4.20-4.67 (3H) .IR (KBr) cm ^-1 : 3400,3230,2920,2850,1490,1467,1
210, 1115, 1090, 1065, 1007. Reference Example 7 3-Hydroxypropylpyridinium tosylate 4.0 g of 1,3-propanediol monotosylate described in Reference Example 1 was dissolved in 10 ml of pyridine and stirred overnight at 60 ° C. The reaction solution was concentrated under reduced pressure to give 3-hydroxypropylpyridinium tosylate as a colorless oil. Yield 5.6 g (yield, quantitative). NMR (90 MHz, DMSO-d ₆ ) δ: 2.07 (2H, quinte
t, J = 7Hz), 2.26 (3H, s), 3.40 (1H, s), 3.43 (2H, t, J = 7Hz),
4.68 (2H, t, J = 7Hz), 7..06 (2H, d, J = 8Hz), 7.48 (2H, d, J =
8Hz), 8.10 (2H, m), 8.56 (1H, m), 9.07 (2H, m). Reference Example 8 2- (benzoyloxy) -3- (octadecyloxy) propyl 3-pyridiniopropyl phosphate chloroform 70 ml , Phosphorus oxychloride 1.62 g (10.5 g
Mmol) and 7.0 ml (52 mmol) of triethylamine under ice cooling, 4.6 g of 1-octadecyl-2-benzoylglycerin synthesized by the method described in Example 8
A 35 ml solution of (10 mmol) in chloroform was added dropwise over 40 minutes. Then, the mixture was stirred at room temperature for 1 hour and cooled with ice, and 3.9 g (12.6 mmol) of pyridine 3 of 3-hydroxypropylpyridinium tosylate obtained in Reference Example 7 was obtained.
The 0 ml solution was added dropwise. After stirring the reaction mixture overnight at room temperature, 7.0 g of a saturated aqueous solution of sodium hydrogen carbonate was added to the mixture, and the mixture was concentrated to dryness under reduced pressure. Toluene 1 in the residue
00 ml and 100 ml of dichloromethane were added, and the insoluble matter was removed by filtration. The filtrate was concentrated under reduced pressure, the residue was dissolved in water, and this was chromatographed on a column having 40 ml of Amberlite IRA-410 and 20 ml of Amberlite IR-120 connected, and eluted with water and then with 95% hydrous tetrahydrofuran. did. The eluate was concentrated and the residue was chromatographed on a column of silica gel (50 g), chloroform-methanol-water (65:25:
4) Elute and concentrate the desired fractions to give 2- (benzoyloxy) -3- (octadecyloxy) propyl.
3-Pyridiniopropyl phosphate was obtained as a colorless solid. Yield 2.4 g (yield 37.1%) Thin layer chromatography [silica gel, chloroform-methanol-water (65: 25: 4)] Rf = 0.23 single spot NMR (90 MHz, CDCl ₃ ) δ: 0.86 (3H ), 1.22 (30H),
1.46 (2H), 2.20 (2H), 3.30 ~ 4.15 (8H), 4.88 (2H), 5.34 (1
H), 7.40 (3H), 7.90 (4H), 8.23 (1H), 9.34 (2H) .IR (KBr) cm ^-1 : 3400,2930,2860,1720,1635,1495,1
465,1285,1240,1100,1070,710. Reference Example 9 2- (Hydroxy) -3- (octadecyloxy) propyl 3-pyridiniopropyl phosphate 2.4 g of 2-benzoyloxy compound synthesized in Reference Example 8 (3
(7 mmol) was dissolved in 5 ml of methanol, 11.6 g (4.45 mmol) of 10% tetrabutylammonium hydroxide aqueous solution was added, and the mixture was stirred at room temperature for 1.5 hours. Amberlite IRA-410 40 ml Amberlite I
Charge 20 ml of R-120 to the connected column and
Elution was performed with% hydrous tetrahydrofuran. The eluate was concentrated to dryness, the residue was chromatographed on a column of silica gel (30 g) and eluted with chloroform-methanol-water (65: 25: 4). Concentrate the desired fraction under reduced pressure,
Acetone was added to the residue, the insoluble material was collected by filtration and dried, and 2- (hydroxy) -3- (octadecyloxy) propyl 3-
Pyridiniopropyl phosphate was obtained as a colorless solid. Yield 1.65 g (Yield 81.9%) Silica gel thin layer chromatography (Merck Company Art 5
715); Rf = 0.11 (chloroform-methanol-
Water = 65: 25: 4) NMR (90 MHz, CDCl ₃ -CD ₃ OD) δ: 0.87 (3
H), 1.24 (30H), 1.50 (2H), 2.30 (2H), 3.40 (4H), 3.90 (5H),
4.84 (2H), 8.07 (2H), 8.43 (1H), 9.15 (2H) .IR (KBr) cm ^-1 : 3350,2925,2850,1635,1490,1470,1
230,1070,785. Reference Example 10 2- (Benzyloxy) -3- (hexadecyloxy)
Propyl 3-trimethylammoniopropyl phosphate 2.3 g (5.7 mmol) of 2-benzyloxy-3-hexadecyloxypropanol and 2.3 g of 3-hydroxypropyltrimethylammonium tosylate obtained in Reference Example 1 in the same manner as in Reference Example 3. From (8.4 mmol), 2- (benzyloxy) -3- (hexadecyloxy) propyl 3-trimethylammoniopropyl phosphate was obtained as a colorless solid product. Yield 2.3 g (yield 69%) Thin layer chromatography [silica gel, chloroform-methanol-water (65: 50: 8)] Rf = 0.18 single spot.

NMR (90 MHz, CDCl ₃ ) δ: 0.87 (3H), 1.25 (26H),
1.45 (2H), 1.98 (2H), 3.06 (9H), 3.26 ~ 4.34 (11H), 4.64 (2
H), 7.30 (5H). Reference Example 11 2- (hydroxy) -3- (hexadecyloxy) propyl 3-trimethylammoniopropyl phosphate 2-benzyl obtained in Reference Example 10 in the same manner as in Reference Example 5. The title compound was obtained as a colorless solid from 2.0 g (3.8 mmol) of the oxy form. Yield 1.6 g (yield 84%) Thin layer chromatography [silica gel, chloroform-methanol-water (65: 50: 8)] Rf = 0.1 single spot NMR (90 MHz, CDCl ₃ + CD ₃ OD) δ: 0.87 (3
H), 1.25 (26H), 1.50 (2H), 2.10 (2H), 3.22 (9H), 3.33 ~ 4.05
(11H). Reference Example 12 2- (benzyloxy) -3- (octadecyloxy)
Propyl 4-bromobutyl phosphate 2-benzyloxy-3-octadecyloxypropanol 5.86 g (13.5 mmol) and 4-bromobutyl
Phosphorodichloridate (4.37 g, 16.2 mmol) was dissolved in dry toluene (35 ml), and the mixture was stirred at room temperature for 30 minutes. Then 1.28 g (16.2 mmol) of dry pyridine was added dropwise. The reaction solution was stirred at room temperature for 18 hours and then concentrated to dryness under reduced pressure. 45 ml of water was added to the residue, and the mixture was heated under reflux for 30 minutes. After standing to cool, the reaction mixture was diluted with 200 ml of dichloromethane.
After extraction, the dichloromethane layer was dried and concentrated under reduced pressure to give the title compound. Yield 8.7 g (yield 99.2%).

NMR (90 MHz, CDCl ₃ -CD ₃ OD) δ: 0.87 (3H),
1.26 (30H), 1.50 (2H), 1.82 ~ 2.05 (4H), 3.32 ~ 3.55 (6H),
3.77 (1H), 4.03 (2H), 4.67 (2H), 7.31 (5H). Reference Example 13 2- (benzyloxy) -3- (octadecyloxy)
Propyl 4- (N-methylpyrrolidinio) butyl phosphate 8.70 g (13.4 mmol) of the compound obtained in Reference Example 12 was dissolved in 100 ml of dry toluene to give N-methylpyrrolidine.
4.56 g (53.6 mmol) was added, and the mixture was stirred at 60 ° C. for 23 hours. The reaction solution was concentrated to dryness under reduced pressure, and the residue was subjected to silica gel column chromatography (Merck, Art. 77).
34, developing solvent, chloroform: methanol: water = 6
5: 25: 4), and 3.75 g of the title compound (yield 4
2.8%).

Silica gel thin layer chromatography (Merck, Art. 5)
715): Rf = 0.35 (chloroform-methanol-
Water = 65: 25: 4) NMR (90 MHz, CDCl ₃ -CD ₃ OD) δ: 0.88 (3
H), 1.27 (30H), 1.50 ~ 2.00 (6H), 2.20 (4H), 2.29 (3H), 3.33
~ 3.60 (10H), 3.75 ~ 4.01 (3H), 4.70 (2H), 7.33 (5H) Reference Example 14 2- (Hydroxy) -3- (octadecyloxy) propyl 4- (N-methylpyrrolidinio) butyl phosphate 3.73 g (5.7 mmol) of the compound obtained in Reference Example 13 was dissolved in a mixed liquid of 20 ml of ethanol and 100 ml of 70% acetic acid, and 2.0 g of 10% Pd / C was added for catalytic reduction. After the reaction, the catalyst was removed and the filtrate was concentrated to dryness under reduced pressure to give the title compound (3.18 g, yield 99.0%).

Thin layer chromatography [silica gel, chloroform-
Methanol-water (65: 25: 4)] Rf = 0.20 single spot NMR (90 MHz, CDCl ₃ -CD ₃ OD) δ: 0.87 (3
H), 1.27 (30H), 1.61 (6H), 2.22 (4H), 3.03 (3H), 3.47 (8H),
3.77 (5H). Reference Example 15 2- (benzyloxy) -3- (octadecyloxy)
Propyl 5- (pyrrolidino) pentyl phosphate Phosphorus oxychloride 2.57 g (16.8 mmol) was dissolved in 3 ml of trichlorethylene, and while stirring this in an ice bath, 1.87 g of 5-bromopentanol (11.2 mmol). Was added. Immediately remove it from the ice bath and let it stand at room temperature for 15
I stirred it for a while. The mixture was concentrated under reduced pressure in a water bath at 40 ° C or lower, 20 ml of toluene was added, and the mixture was concentrated again.

The residue was dissolved in 20 ml of toluene, and 3 g (7 mmol) of 2-benzyloxy-3-octadecyloxypropanol and 1.8 g of pyridine were added thereto while stirring at room temperature. The mixture was stirred at room temperature for 1.5 hours and toluene was distilled off under reduced pressure. 40 ml of water was added to the residue and the mixture was heated under reflux for 2 hours.
After cooling the reaction mixture, it was partitioned with 80 ml of dichloromethane, the dichloromethane was distilled off, the residue was dissolved in 40 ml of ethanol, and 3.8 g (62.3 mmol) of pyrrolidine was added and the mixture was heated to 80 ° C. at 2.5.
After stirring for an hour, the mixture was dried under reduced pressure. The residue was partitioned between water and dichloromethane, the dichloromethane was distilled off, and the residue was purified by silica gel column chromatography (methanol),
2.3 g (50% yield) of the title compound was obtained.

Silica gel thin layer chromatography (Merck, Art. 5)
715) Rf = 0.35 (methanol), single spot NMR (60 MHz, CDCl ₃ -CD ₃ OD) δ: 0.90 (3
H), 1.27 (32H), 1.50 ~ 1.80 (6H), 1.90 ~ 2.27 (4H), 2.73 ~
3.17 (6H), 3.30~4.17 (9H ), 4.70 (2H), 7.33 (5H) IR (CHCl 3) cm -1: 2450,1090,1050,1000 Reference Example 16 2- (hydroxymethyl) -3- (octadecyl Oxy) propyl 5- (pyrrolidino) pentyl phosphate 2- (benzyloxy) -3- (octadecyloxy)
Propyl 5- (pyrrolidino) pentyl phosphate 2.
3 g was dissolved in 40 ml of ethanol, 1.2 g of 10% palladium-carbon was added, and catalytic reduction was carried out at room temperature under normal pressure in a hydrogen stream. The catalyst was removed by filtration, and the filtrate was concentrated under reduced pressure to give the title compound (1.75 g, yield 90%).

Silica gel thin layer chromatography (Merck, Art. 5)
715) Rf = 0.46 (chloroform: methanol: water = 65: 25: 4), single spot NMR (60 MHz, CDCl ₃ -CD ₃ OD) δ: 0.90 (3H),
1.27 (32H), 1.70 ~ 1.87 (6H), 2.00 ~ 2.30 (4H), 2.87 ~ 3.27
(6H), 3.33 to 3.67 (4H), 3.87 to 4.20 (5H) IR (CHCl ₃ ) cm ^-1 : 3340,2575,2450,1225,1205,110
0,1010 Example 1 2- (acetoacetyloxy) -3- (octadecyloxy) propyl 2-trimethylammonioethyl phosphate 2-hydroxy-3- (octadecyloxy) propyl 2-trimethylammonioethyl phosphate 1.0
g (1.96 mmol) was dissolved in a mixed solvent of 20 ml of pyridine and 20 ml of dichloromethane, and 3 ml of diketene was added dropwise over 30 minutes while stirring at 40 ° C. The reaction solution was evaporated, the residue was purified by column chromatography using 15 g of silica gel and an eluent (chloroform: methanol: water = 65: 25: 4), and the target fraction was concentrated to dryness. . Acetone was added to the residue for solidification to obtain the desired product as a pale yellow powder. Yield 810 mg (yield 69.4%).

Silica gel thin layer chromatography (Merck, Art. 5)
715): Rf = 0.51 (chloroform: methanol:
Water = 65: 25: 4). NMR (90 MHz, CDCl ₃ ) δ: 0.81 (3H), 1.25 (30H),
1.50 (2H), 2.26 (3H), 3.39 (9H), 3.50 (2H), 3.33 ~ 4.30 (10
H), 5.20 (1H). Example 2 (S) -2-acetoacetyloxy-3- (octadecyloxy) propyl 2-trimethylammonioethyl
Phosphate (S) -2-hydroxy-3- (octadecyloxy) propyl 2-trimethylammonioethyl phosphate 928 mg (1.8 mmol) and diketene 1 ml were treated in the same manner as in Example 1 to obtain 827 mg (77%) of the title compound. It was

Optical rotation: [α] ▲ ^{25 °} _D ▼ = -0.25 ° (c = 1.59,
Methanol) Example 3 2-acetoacetyloxy-3- (octadecyloxy) propyl 2-dimethylaminoethyl phosphate 2-benzyloxy-3- (octadecyloxy) propyl 2-bromoethyl phosphate 7.5 g (12 mmol) 12.8 % Alcoholic dimethylamine 42g
It was dissolved in (120 mmol) and left at room temperature for 7 days.
The solvent was distilled off under reduced pressure, the residue was dissolved in 60 ml of methanol, 3.3 g (12 mmol) of silver carbonate was added, and the mixture was heated under reflux for 1 hour. The insoluble matter was removed by filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform: methanol: water = 65: 25: 4) to give 2-
Benzyloxy-3- (octadecyloxy) propyl 2-dimethylaminoethyl phosphate 5.2 g (7
4%).

Silica gel thin layer chromatography (Merck, Art. 5)
715): Rf = 0.57 (chloroform: methanol:
Water = 65: 25: 4) NMR (60 MHz, CDCl ₃ ) δ: 0.87 (3H), 1.23 (32
H), 2.70 (6H), 3.30 (1H), 3.30 to 4.33 (10H), 4.63 (2H), 7.23
(5H). Dissolve 5.2 g of the above compound in 100 ml of ethanol and prepare 5%.
Palladium-carbon (2 g) was added, and the mixture was catalytically reduced in a hydrogen stream at room temperature and atmospheric pressure. The catalyst was removed by filtration, and the filtrate was concentrated under reduced pressure to give 2-hydroxy-3- (octadecyloxy) propyl 2-dimethylaminoethyl phosphate 3.1 g.
(70%) was obtained.

Silica gel thin layer chromatography (Merck, Art. 5)
715): Rf = 0.41 (chloroform: methanol:
Water = 65: 25: 4).

NMR (60 MHz, CDCl ₃ -CD ₃ OD) δ: 0.90 (3
H), 1.27 (32H), 2.90 (6H), 3.20-4.33 (11H). 1.49 g (3 mmol) of the above compound was added to 25 ml of pyridine, and 840 mg (10 mmol) of diketene was added to give 5
Stir vigorously for 1 hour at 0 ° C. n-propanol 2
0 ml was added to the reaction solution, the reaction solution was concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography (chloroform:
Purification with methanol: water = 65: 25: 4) yielded 1.5 g (86%) of the title compound.

Silica gel thin layer chromatography (Merck, Art. 5)
715): Rf = 0.52 (chloroform: methanol:
Water = 65: 25: 4) IR (CHCl ₃ ) cm ⁻¹ : 2930,2860,2470,1740,1715,146
5,1235,1085,1050. NMR (60MHz, CDCl ₃ ) δ: 0.87 (3H), 1.23 (32H),
2.27 (3H), 2.87 (6H), 3.10-4.40 (13H), 5.07-5.40 (1H). Example 4 2-acetoacetyloxy-3- (octadecyloxy) propyl 2-pyrrolidinoethyl phosphate 2-benzyloxy 7.5 g (12 mmol) of 3- (octadecyloxy) propyl 2-bromoethyl phosphate and 8.5 g (120 mmol) of pyrrolidine were dissolved in 30 ml of toluene and stirred at 60 ° C. for 18 hours. The solvent was distilled off under reduced pressure, and the residue was dissolved in 60 ml of methanol,
3.3 g (12 mmol) of silver carbonate was added and the mixture was heated under reflux for 1 hour. The insoluble material was removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform: methanol: water = 65: 25: 4) to give 2
4.94 g (68%) of -benzyloxy-3- (octadecyloxy) propyl 2-pyrrolidinoethyl phosphate were obtained.

Silica gel thin layer chromatography (Merck, Art. 5)
715): Rf = 0.68 (chloroform: methanol:
Water = 65: 25: 4) NMR (60 MHz, CDCl ₃ ) δ: 0.87 (3H), 1.23 (32H),
1.97 (4H), 2.93 to 4.33 (15H), 4.67 (2H), 7.27 (5H). 4.94 g of the above compound was added to ethyl acetate-ethanol (1:
1) Dissolved in 100 ml, added 5 g of 5% palladium-carbon, and catalytically reduced in a hydrogen stream at room temperature under normal pressure. The catalyst was removed by filtration, and the filtrate was evaporated under reduced pressure to give 2-hydroxy-3-
3.4 g (81%) of (octadecyloxy) propyl 2-pyrrolidinoethyl phosphate were obtained.

Silica gel thin layer chromatography (Merck, Art. 5)
715): Rf = 0.52 (chloroform: methanol:
Water = 65: 25: 4) NMR (60 MHz, CDCl ₃ ) δ: 0.87 (3H), 1.23 (32H),
2.07 (4H), 3.10-4.47 (15H) 887 mg (1.7 mmol) of the above compound, 840 mg of diketene
(10 mmol) was added to 20 ml of pyridine, and the mixture was vigorously stirred at 50 ° C. for 1 hour. 20 ml of n-propanol was added to the reaction solution, which was concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (chloroform: methanol:
After purification with water = 65: 25: 4), 742 mg of the title compound
(73%) was obtained.

Silica gel thin layer chromatography (Merck, Art. 5)
715): Rf = 0.55 (chloroform: methanol:
Water = 65: 25: 4) IR (CHCl ₃ ) cm ⁻¹ : 2930,2860,2480,1740,1715,146
NMR (60 MHz, CDCl ₃ ) δ: 0.90 (3H), 1.27 (32H),
2.10 (4H), 2.30 (3H), 3.03 to 4.43 (17H), 4.97 to 5.43 (1H). Example 5 2-acetoacetyloxy-3- (octadecyloxy) propyl 2- (N-methylpyrrolidinio) 7.5 g (12 mmol) of ethyl phosphate 2-benzyloxy-3- (octadecyloxy) propyl 2-bromoethyl phosphate and 10.2 g (120 mmol) of N-methylpyrrolidine were dissolved in 30 ml of toluene and left at room temperature for 8 hours. . The solvent was distilled off under reduced pressure, and the residue was added with 60 ml of methanol.
The solution was dissolved in, and 3.3 g (12 mmol) of silver carbonate was added, and the mixture was heated under reflux for 1 hour. The insoluble material was removed by filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform: methanol: water = 65: 25: 4) to give 2-benzyloxy-3- (octadecyloxy).
3.5 g (47%) of propyl 2- (N-methylpyrrolidinio) ethyl phosphate was obtained.

Silica gel thin layer chromatography (Merck, Art. 5)
715): Rf = 0.49 (chloroform: methanol:
Water = 65: 25: 4) NMR (60 MHz, CDCl ₃ ) δ: 0.90 (3H), 1.27 (32H),
2.03 (4H), 3.07 (3H), 3.30 ~ 4.40 (15H), 4.67 (2H), 7.27 (5
H). 3.5 g of the above compound was dissolved in 60 ml of ethanol, 1.5 g of 5% palladium-carbon was added, and catalytic reduction was carried out in a hydrogen stream at room temperature under normal pressure. The catalyst was removed by filtration, and the filtrate was evaporated under reduced pressure to give 2.57 g (86%) of 2-hydroxy-3- (octadecyloxy) propyl 2- (N-methylpyrrolidinio) ethyl phosphate.

Silica gel thin layer chromatography (Merck, Art. 5)
715): Rf = 0.36 (chloroform: methanol:
Water = 65: 25: 4).

NMR (60 MHz, CDCl ₃ -CD ₃ OD) δ: 0.90 (3
H), 1.27 (32H), 2.03 (4H), 3.13 (3H), 3.30-4.40 (15H). 911 mg (1.7 mmol) of the above compound and 840 mg of diketene.
(10 mmol) was added to 40 ml of pyridine, and 1 at 50 ° C.
The time was agitated. 20 ml of n-propanol was added to the reaction solution, which was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform: methanol: water = 65: 25: 4) to give 665 mg of the title compound.
(62%) was obtained.

Silica gel thin layer chromatography (Merck, Art. 5)
715): Rf = 0.49 (chloroform: methanol:
Water = 65: 25: 4). IR (CHCl ₃ ) cm ⁻¹ : 2925,2860,1710,1460,1070,104
0. NMR (60 MHz, CDCl ₃ ) δ: 0.90 (3H), 1.27 (32H),
2.23 (4H), 2.27 (3H), 3.27-4.60 (20H), 4.93-5.43 (1H). Example 6 2- (acetoacetyloxy) -3- (hexadecyloxy) propyl 2-trimethylammonioethyl phosphate 2-Hydroxy-3- (hexadecyloxy) propyl 2-trimethylammonioethyl phosphate 1.3
5 g (2.8 mmol) was added to 40 ml of pyrrolidine, 1.74 g (20 mmol) of diketene was added, and the mixture was vigorously stirred at 50 ° C. for 1 hour. 20 ml of n-propanol was added to the reaction solution, the reaction was concentrated under reduced pressure, the residue was purified by silica gel column chromatography (chloroform: methanol: water = 65: 25: 4) and solidified by adding acetone to give the title compound 1 0.11 g (70%) was obtained.

Silica gel thin layer chromatography (Merck, Art. 5)
715): Rf = 0.36 (chloroform: methanol:
Water = 65: 25: 4).

IR (CHCl ₃ ) cm ^-1 : 2930,2860,1745,1715,1250,109
NMR (60 MHz, CDCl ₃ ) δ: 0.90 (3H), 1.27 (28H),
2.27 (3H), 3.10 to 4.50 (22H), 4.87 to 5.27 (1H). Example 7 2-acetoacetyloxy-3- (tetradecyloxy) propyl 2-trimethylammonioethyl phosphate 2-hydroxy-3- ( Hexadecyloxy) propyl 2-trimethylammonioethyl phosphate 1.3
Add 6 g (3 mmol) to 40 ml of pyridine and add diketene.
1.74 g (20 mmol) was added and the mixture was vigorously stirred at 50 ° C. 20 ml of n-propanol was added to the reaction solution, the reaction solution was concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography (chloroform: methanol: water = 65: 2).
After purification by 5: 4) and addition of acetone to solidify, 1.11 g (69%) of the title compound was obtained.

Silica gel thin layer chromatography (Merck, Art. 5)
715): Rf = 0.35 (chloroform: methanol:
Water = 65: 25: 4) IR (CHCl ₃ ) cm ⁻¹ : 2930,2860,1740,1720,1255,109
NMR (60 MHz, CDCl ₃ ) δ: 0.90 (3H), 1.27 (24H),
2.30 (3H), 3.13-4.47 (22H), 4.97-5.40 (1H). Example 8 2-acetoacetyloxy-3- (octadecyloxy) propyl 2-pyrrolidinioethyl phosphate 1-trityl-3-octadecylglycerin 32.0 g
(53.4 mmol) was dissolved in 200 ml of dry dichloromethane, and stirred with stirring at 0 to 3 ° C. to obtain dry pyridine 22.4.
Then a solution of 6.7 ml (57.9 mmol) of benzoyl chloride-100 ml of dry dichloromethane was added dropwise over 30 minutes. After stirring the reaction mixture at room temperature for another 2 hours,
The reaction solution was concentrated under reduced pressure. 200 ml of ether was added to the residue, and the insoluble material was filtered off. After concentrating the filtrate, the residue was subjected to silica gel column chromatography (hexane-ethyl acetate =
9: 1) to give 2-benzoyl-3-octadecyl-1-tritylglycerin as an oil, 28.8 g (7
%)Obtained.

28.8 g (41 mmol) of the above compound and 20 ml of 1N hydrochloric acid
Was added to 400 ml of dioxane, and the mixture was stirred at 80 ° C. for 30 minutes. The reaction solution was neutralized by adding saturated aqueous sodium hydrogen carbonate under ice-cooling, and the solvent was evaporated under reduced pressure. Dichloromethane was added to the residue, and the insoluble material was filtered off. The filtrate was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform) to give an oily 1
-Octadecyl-2-benzoylglycerin 17.15g
(93%) was obtained.

8.98 g (20 mmol) of the above compound and 7.26 g (30 mmol) of 2-bromoethyl phosphoric acid dichloride were dissolved in 50 ml of dry toluene and stirred at room temperature for 30 minutes, and then 2.38 g (30 mmol) of dry pyridine was added. Dropped. After stirring at room temperature for another 1.5 hours, pyridine 2.3
8 g and 10 ml of water were added, and the mixture was stirred overnight at room temperature. After separating the organic layer, the aqueous layer was extracted with toluene, and the organic layers were combined and filtered. The filtrate was evaporated under reduced pressure, and the residue was subjected to silica gel column chromatography (chloroform: methanol:
Purification with water = 65: 2: 4) gave 8.83 g (69%) of oily 2-benzoyloxy-3- (octadecyloxy) propyl 2-bromoethyl phosphate.

The above compound (1.36 g, 5.66 mmol) was dissolved in dry pyridine (36 ml), and the mixture was stirred at 50 ° C. overnight. 60 more
After stirring at 24 ° C. for 24 hours, the reaction solution was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform: methanol: water = 65: 25: 4) to obtain 2.56 g (71%) of oily 2-benzoyloxy-3- (octadecyloxy) propyl-2-pyridinioethyl phosphate. Obtained.

2.22 g (3.5 mmol) of the above compound was added to 10 m of methanol.
Dissolve in 1 l, add 10.9 g (4.2 mmol) of 10% tetra-n-butylammonium hydroxide aqueous solution, and add 4.5 at room temperature.
Stir for hours. The reaction solution was purified by XAD-II column chromatography (elution with water and methanol in this order), and the fractions containing the desired product were concentrated under reduced pressure. Residue 1.57
When g was purified by silica gel column chromatography (chloroform: methanol: water = 65: 25: 4), solid 2-hydroxy-3- (octadecyloxy) propyl 2-pyridinioethyl phosphate 1.06 g was obtained.
(57%) was obtained.

100 mg (0.19 mmol) of the above compound was dissolved by heating in a mixed solution of 2 ml of dry pyridine and 2 ml of dry dichloromethane,
0.5 ml of diketene was added dropwise at 25 to 35 ° C. 23-38
After stirring at 30 ° C. for another 30 minutes, the mixture was concentrated under reduced pressure.
4 ml of acetone was added to the residue, and the mixture was left overnight at room temperature. The solid matter is collected by filtration, washed with a small amount of acetone, and then dried under reduced pressure (on anhydrous phosphoric acid) to give 2-acetoacetyloxy-3.
59 mg (51%) of-(octadecyloxy) propyl 2-pyridinioethyl phosphate were obtained.

Silica gel thin layer chromatography (Merck, Art.
5715): Rf = 0.24 (chloroform: methanol: water = 65: 25: 4) NMR (90 MHz, CDCl ₃ ) δ: 0.86 (3H), 1.25 (30H),
1.47 (2H), 2.20 (3H), 3.29 ~ 3.53 (4H), 3.45 (2H), 3.88 (2
H), 4.31 (2H), 5.01 (2H), 5.14 (1H), 8.05 (2H), 8.43 (1H), 9.
28 (2H). Example 9 2- (acetoacetyloxy) -3- (octadecyloxy) propyl 3-trimethylammoniopropyl
Phosphate 800 mg of the 2-hydroxyl product obtained in Reference Example 5 was dissolved in a mixed solution of 30 ml of pyridine and 10 ml of dichloromethane, and 2 ml of diketene was added while stirring at 40 ° C. Two
After a while, the solvent was distilled off from the reaction solution, the residue was chromatographed on a column of silica gel (15 g), and eluted with chloroform-methanol-water (65: 25: 4) to obtain a desired fraction. Obtained. This is concentrated under reduced pressure, acetone is added to the residue to solidify it, and 2- (acetoacetyloxy)
-3- (Octadecyloxy) propyl 3-trimethylammoniopropyl phosphate was obtained as a pale yellow solid. Yield 730 mg (Yield 77%) Thin layer chromatography [silica gel, chloroform-methanol-water (65: 25: 4) Rf = 0.13 single spot NMR (90 MHz, CDCl ₃ ) δ: 0.87 (3H), 1.26 ( 30H),
1.53 (2H), 2.13 (2H), 2.26 (3H), 3.30 (9H), 3.40-3.70 (6
H), 3.53 (2H), 3.83 to 4.03 (4H), 5.20 (1H) .IR (KBr) cm ^-1 : 3420,2920,2840,1740,1715,1465,1
235,1090,1055,840. Example 10 2- (acetoacetyloxy) -3- (octadecyloxy) propyl 5-trimethylammoniopentyl
Phosphate 800 mg (1.5 mg) of 2-hydroxyl compound obtained in Reference Example 6
Was treated in the same manner as in Example 9 to obtain the desired product as a pale yellow solid. Yield 750 mg (81% yield) Thin layer chromatography [silica gel, chloroform-methanol-water (65: 25: 4)] Rf = 0.15 single spot NMR (90 MHz, CDCl ₃ ) δ: 0.90 (3H), 1.26 (30H),
1.46 (2H), 1.60 (4H), 1.90 (2H), 2.26 (3H), 3.30 (9H), 3.40
−3.63 (6H), 3.46 (2H), 3.83−4.00 (4H), 5.23 (1H) .IR (KBr) cm ⁻¹ : 3400,2920,2850,1740,1715,1665,1
235,1090,1070,835. Example 11 2- (acetoacetyloxy) -3- (octadecyloxy) propyl 3-pyridiniopropyl phosphate 1.3 g of 2-hydroxyl compound obtained in Reference Example 9 (2.39)
(Mmol) was treated in the same manner as in Example 9 to obtain the desired product as a yellow solid. Yield 930 mg (yield 62.1%).

Thin layer chromatography [silica gel, chloroform-methanol-water (65: 25: 4)] Rf = 0.20 single spot NMR (90 MHz, CDCl ₃ -CD ₃ OD) δ: 0.87 (3
H), 1.26 (30H), 1.52 (2H), 2.26 (3H), 2.27 (2H), 3.33 ~ 4.03
(8H), 3.71 (2H), 4.82 (2H), 5.19 (1H), 8.06 (2H), 8.47 (1H),
9.10 (2H). IR (KBr) cm ^-1 : 3400,2920,2850,1735,1715,1630,1
490,1460,1230,1090,1060,970,840,810. Example 12 2- (acetoacetyloxy) -3- (hexadecyloxy) propyl 3-trimethylammoniopropyl
Phosphate 800 mg of 2-hydroxyl compound obtained in Reference Example 11 (1.
(6 mmol) was treated in the same manner as in Example 9 to obtain the title compound as a pale yellow solid. Yield 650 mg (yield 70
%).

Thin layer chromatography [silica gel, chloroform-methanol-water (65: 25: 4)] Rf = 0.12 single spot NMR (90 MHz, CDCl ₃ ) δ: 0.87 (3H), 1.25 (26H),
1.50 (2H), 2.12 (2H), 2.26 (3H), 3.30 (9H), 3.40 ~ 4.03 (10
H), 3.51 (2H), 5.21 (1H) .IR (KBr) cm ^-1 : 3420,2920,2850,1740,1715,1465,1
235,1090,1060,840. Example 13 2- (acetoacetyloxy) -3- (octadecyloxy) propyl 4- (N-methylpyrrolidinio) butyl phosphate Compound 2.00 g (3 0.55 mmol) was dissolved in a mixed solution of 20 ml of dry dichloromethane and 40 ml of dry pyridine, 4 ml of diketene was added, and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated to dryness under reduced pressure, and the residue was subjected to silica gel column chromatography (Merck, Art. 773).
4, developing solvent, chloroform: methanol: water = 65:
25: 4), and 1.45 g of the title compound (yield 63.2).
%) Was obtained.

Thin layer chromatography [silica gel, chloroform-
Methanol-water (65: 25: 4)] Rf = 0.30 single spot NMR (90 MHz, CDCl ₃ -CD ₃ OD) δ: 0.86 (3
H), 1.26 (30H), 1.50 ~ 2.03 (6H), 2.27 (7H), 3.04 (3H), 3.33
~ 3.63 (6H), 3.79 ~ 4.03 (4H), 5.18 (1H) IR (KBr) cm ^-1 : 3425,2920,2855,1740,1715,1650,1
465,1230,1065,825. Example 14 2- (acetoacetyloxy) -3- (octadecyloxy) propyl 5- (pyrrolidino) pentyl phosphate 2- (hydroxy) -3- (octadecyloxy) propyl 5- (pyrrolidino ) Pentyl phosphate 1.7g
(3 mmol) was dissolved in 20 ml of pyridine and 3.0 of diketene was added.
g (35.7 mmol) was added, and the mixture was stirred at 50 ° C for 0.5 hr. After adding 30 ml of ethanol and concentrating under reduced pressure, the residue was subjected to silica gel column chromatography (chloroform: methanol: water = 65: 25: 1 to 65:25:
Purification in 4) yielded 1.3 g of the title compound (yield 67%).

Silica gel thin layer chromatography (Merck, Art.
5715) Rf = 0.51 (chloroform: methanol:
Water = 65: 25: 4) single spot NMR (60 MHz, CDCl ₃ -CD ₃ OD) δ: 0.90 (3
H), 1.27 (32H), 1.53 ~ 1.87 (6H), 2.00 ~ 2.27 (4H), 2.33 (3
H), 2.83 to 3.27 (6H), 3.43 to 4.33 (9H), 5.07 to 5.40 (1H) IR (CHCl ₃ ) cm ^-1 : 2460,1740,1715,1230,1200,1050 Example 15 2- (aceto Acetyloxy) -3- (octadecyloxy) propyl 5- (N-methylpyrrolidinio) pentylphosphate 2- (acetoacetyloxy) -3- (octadecyloxy) propyl 5- (pyrrolidino) pentylphosphate 1.17 g (1.8 820 mg (10 mmol) of sodium bicarbonate dissolved in 30 ml of acetone and ground in a mortar.
Mmol) and 410 mg of methyl para-toluenesulfonate
(2.2 mmol) was added and the mixture was stirred at 50 ° C. for 13 hours. Acetone was distilled off under reduced pressure, the residue was dissolved in 30 ml of water, adjusted to pH 4 with 2N HCl, and then extracted with a mixed solvent of dichloromethane: ethanol 10: 1. The solvent was distilled off and the residue was purified by silica gel column chromatography (chloroform: methanol: water = 65: 25: 4) to obtain 550 mg (yield 46%) of the title compound.

Silica gel thin layer chromatography (Merck, Art.
5715) Rf = 0.25 (chloroform: methanol:
Water = 65: 25: 4) Single spot NMR (60 MHz, CDCl ₃ -CD ₃ OD) δ: 0.87 (3
H), 1.23 (32H), 1.50 ~ 1.80 (6H), 2.20 ~ 2.40 (4H), 2.30 (3
H), 3.07 (3H), 3.23 ~ 3.73 (11H), 3.83 ~ 4.10 (4H), 5.07 ~
5.40 (1H) IR (CHCl ₃ ) cm ⁻¹ : 1735,1715,1235,1200,1090,1065 Example 16 2- (acetoacetyloxy) -3- (octadecyloxy) propyl 4-trimethylammoniobutyl phosphate Reference The title compound was obtained as a yellow solid in the same manner as in Examples 1, 3, 5 and Example 9.

Thin layer chromatography [silica gel, chloroform-methanol-water (65: 25: 4)] Rf = 0.30 single spot NMR (90 MHz, CDCl ₃ -CD ₃ OD) δ: 0.87 (3
H), 1.27 (30H), 1.45-2.02 (6H), 2.27 (3H), 3.10 (9H), 3.43
(4H), 3.61 (2H), 3.91 (6H), 5.20 (1H) IR (KBr) cm ^-1 : 3410,2920,2850,1740,1715,1630,1
465, 1225, 1090, 1065, 830 The following compounds can be produced in the same manner as in the above Reference Examples and Examples.

2- (Acetoacetyloxy) -3- (octadecyloxy) propyl 3-pyrrolidinopropyl phosphate 2- (acetoacetyloxy) -3- (octadecyloxy) propyl 3-N-methylpyrrolidiniopropyl phosphate 2- (Acetoacetyloxy) -3- (octadecyloxy) propyl 3-thiazolyopropylphosphate 2- (acetoacetyloxy) -3- (octadecyloxy) propyl 4-pyridiniobutyl phosphate 2- (acetoacetyloxy) -3- (Octadecyloxy) propyl 4-pyrrolidinobutyl phosphate 2- (acetoacetyloxy) -3- (octadecyloxy) propyl 5-pyridiniopentyl phosphate 2- (acetoacetyloxy) -3- (heptadecyl Oxy) propyl 2-trimethylammonioethyl phosphate 2- (acetoacetyloxy) -3- (nonadecyloxy) propyl 3-trimethylammoniopropyl phosphate Formulation Example 1 50 g of the compound of Example 1 was dissolved in 1.0 distilled water, After aseptic filtration, 1 ml is dispensed into 1000 vials under aseptic conditions, freeze-dried, and sealed after drying.

Separately, 2 ml of distilled water for injection containing 100 g of xylit or mannitol is aseptically dispensed in 2 ml aliquots into ampules for injection and then sealed to prepare 1000 bottles.

At the time of use, the former 1 vial of powder is dissolved in xylit solution for injection (or mannitol solution) and used.

Formulation Example 2 As a usage amount per tablet (1) Compound of Example 3 100 mg (2) Lactose 200 mg (3) Corn starch 51 mg (4) Hydroxypropyl cellulose 9 mg are mixed and granulated by a conventional method, and corn starch (8 m
g), and after mixing with magnesium stearate (2 mg), the mixture is tableted to give a tablet of 370 mg and a diameter of 9.5 mm.

Formulation Example 3 Hydroxypropylmethylmethylcellulose phthalate (14
(mg) and castor oil (1 mg) are dissolved at a concentration of 7% to prepare an enteric coated tablet by applying a coating using an acetone-ethanol (4: 6) mixture.

Effect of the invention Test Example 1 Antitumor effect of 2- (acetoacetyloxy) -3- (octadecyloxy) propyl 2-trimethylammonioethyl phosphate (Example 1) 1 × per mouse for ICR mice (5 mice per group) 10 ⁵ Sarcoma 180 cells were transplanted intraperitoneally. Then,
The compound of Example 1 (0.33 mg / mouse) dissolved in physiological saline was intraperitoneally administered intraperitoneally three times at 1 hour, 1 day, and 2 days later. The control compound (IIIa) was also administered under the same conditions.
Table 1 shows the life-prolonging rate and the number of survivors on the 60th day after the start of the test for the dead mice relative to the control group to which no drug was administered.

Test Example 2 Drug 0.25 mg / mouse was used as C3H / He mouse (1 group 5
The mice were intraperitoneally administered daily for 4 days. Mouse 1 on day 6
1 × 10 ⁴ MM46 cells per animal were intraperitoneally transplanted, and 0.25 mg / mouse of the drug was intraperitoneally administered again for 4 days from the second after the transplantation. Table 2 shows the life prolongation rate and the number of surviving animals on the 46th day after transplantation of MM46 regarding the dead mice in the drug administration group to the control group not administered with the drug.

Test Example 3 The antitumor effect of the drug was measured by the same method as in Test Example 2. Table 3 shows the life-prolonging rate and the number of surviving mice on the 47th day after transplantation of MM46 regarding dead mice in the drug-administered group relative to the control group to which the drug was not administered.

Test Example 4 Antitumor activity of the compound of Example 1 ICR mice (5 mice per group) were subcutaneously transplanted with 1 × 10 ⁶ Sarcoma 180 cells per mouse. 8 after transplant
Sun, 9th, 10th, 13th, 14th, 15th, 16th,
After 17 days and 20 days, 0.1 mg / mouse and 0.3 mg / mouse of the compound of Example 1 dissolved in physiological saline were intravenously administered 9 times in total. The control compound (IIIa) 0.3 mg / mouse was administered under the same conditions. Twenty-one days later, the tumor tissue was excised and the tumor weight was measured. Table 4 shows the tumor growth inhibition ratio (comparison with that of the control group to which no drug was administered).

Test Example 5 Action on Platelets [Test Method and Results] Blood was collected from male rabbits using a syringe containing 3.15% citric acid (1 ratio to 9 blood) as an anticoagulant, and at room temperature. Platelet-rich plasma (PRP: Platelet Rich Plasm) by centrifugation at 1000 rpm for 10 minutes.
got a). PRP was further centrifuged at 1400 rpm for 15 minutes to obtain Platelet pellet, which was Ca ₊₊ free T
Suspended in yrode (containing 0.25% gelatin), Washed P
The RP was adjusted. This Washed PRP 250μ is 37
After stirring at 0 ° C. for 2 minutes, 0.2 to 0.5 mM Ca ⁺⁺ solution, 25 μ was added, and the mixture was further stirred for 30 seconds. Then, the test drug was added in an amount to give a predetermined concentration. Platelet aggregation was measured with an aggregometer (manufactured by Rika Denki). The results are shown in Table 5.

Test Example 6 Antihypertensive action Male Sprague-Dawley rats (200 to 290) of 7 weeks old
g) Pentobarbital sodium salt, 60 mg / kg
Was intraperitoneally administered for anesthesia, and the left carotid artery (blood pressure measurement) and the left femoral vein (for intravenous administration) were cannulated.

A predetermined amount of the test compound was administered and blood pressure decrease (ΔmmHg) was measured. The results are shown in Table 5.

Test Example 7 The antitumor effect of the drug was measured under the same conditions as in Test Example 1. Table 6 shows the life extension rate and the number of survivors 60 days after the start of the test with respect to the control group to which no drug was administered.

Test Example 8 By the same test method as in Test Example 5, the platelet aggregation action of the drug was measured, and the concentration of the drug exhibiting 50% platelet aggregation action was calculated. The results are shown in Table 7.

Test Example 9 C3H / He mice (5 mice per group) 1 × 1 per mouse
0 ^four MM46 cells were transplanted into the abdominal cavity. From the second day after the transplantation, 0.25 mg of the drug / mouse was intraperitoneally administered for 4 days. Table 8 shows the life prolongation rate of the dead mice in the drug-administered group relative to the control group not administered with the drug, and the number of surviving animals 60 days after transplantation of MM46.

Claims (1)

[Claims] 1. A formula [Wherein R ¹ represents alkyl, R ² , R ³ and R ⁴ each represent hydrogen or lower alkyl, or Represents a cyclic ammonio group which may have N, S or O as a constituent atom and which may have a substituent, and A represents a lower alkylene] Alternatively, an antitumor agent containing a salt thereof.