JPS58164596A - Phospholipid derivative, its preparation and antitumor agent containing the same - Google Patents

Abstract

NEW MATERIAL:A compound of formulaI(R<1> is alkoxyl; R<2> is H, lower alkyl or aryl; A is lower alkylene; formula II is 6-membered nitrogen-containing heterocyclic group). EXAMPLE:Racemic-1-O-octadecyl-2-O-methylcarbamoyl-glycerine-3- 2-(1-pyridino) ethyl -phosphate. USE:An antitumor agent. PROCESS:A compound of formula III (R<3> is a protecting group of the hydroxy group), e.g. racemic-1-O-octadecyl-3-O-trityl-glycerine, is reacted with an isocyanate of the formula OCNR<2>, e.g. methyl isocyanage, in the presence of a base, e.g. dibutyltin dichloride, to give a compound of formula IV, which is then subjected to the elimination reaction of the protecting group of the hydroxy group to afford a compound of formula V. The resultant compound of formula V is then reacted with a compound of formula VI (X is an acid residue), e.g. 2- bromoethyl dichlorophosphate, to give a compound of formula VII, which is further reacted with a 6-membered nitrogen-containing heterocyclic compound, e.g. pyridine.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to phospholipid derivatives, and more particularly to a novel phospholipid derivative having antitumor activity, a method for producing the same, and a pharmaceutical formulation containing the same.

The phospholipid derivative targeted by this invention is a new compound and is represented by the following general formula (+).

[In the formula, R is an alkoxy group; R is a hydrogen atom, a lower alkyl group, or an aryl group; A is a lower alkylene group; group-0
means a nitrogen-containing 6-membered heterocyclic group optionally substituted with a lower alkyl group] In the above description and the following description of this specification, appropriate definitions of various definitions included within the scope of this invention will be used. Examples and explanations are detailed below.

"Lower" means 1 to 1 carbon atom, unless otherwise specified.
6 and "higher" shall mean 7 to 25 carbon atoms.

Preferred "alco,oxy groups" of H1 are linear or branched alkoxy containing 1 to 25 carbon atoms, methoxy, ethoxy, propoxy, impropoxy, butoxy, tertiary butoxy, pentyloxy, Hexyloxy, hexyloxy, octyloxy, nonyloxy, decyloxy, undecyloxy, dodecyloxy, tocosyloxy, tetradecyloxy, pentadecyloxy, hexadecyloxy, heptadecyloxy, octadecyloxy, nonadecyl oxine, eicosyloxy, hexadecyloxy , tocosyloxy, tocosyloxy,
Examples include tetradecyloxy, bentacyloxy, etc. Among these, particularly preferred alkoxy for R1 is higher alkoxy.

Suitable "lower alkyl group" for R2 is a linear or branched alkyl group containing 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, 6th-butyl, pentyl. , hexyl, etc.

Suitable "aryl groups" for R2 include phenyl, tolyl,
Includes xylyl, mesityl, cumenyl, naphthyl, etc.

A preferred "lower alkylene group" is a linear or branched alkylene containing 2 to 6 carbon atoms, and includes ali, ethylene, trimethylene, propylene, tetramethylene, pentamethylene, hexamethylene, and the like.

The "nitrogen-containing 6-membered heterocyclic group" represented by the group -O is a saturated or unsaturated 6-membered heterocyclic group having 71 to 2 nitrogen atoms in the ring, and preferable examples include pyridinio,
Examples include pyrimidinio, pyridacinio, pyrazinio, piperazino, 1-piperazinyl and the like. These nitrogenous 6
The membered heterocyclic group may be substituted at any position with a lower alkyl group as described above.

The target compound m can also be expressed in the form of an inner salt; for example, when the group -0 in compound [1] is an unsaturated nitrogen-containing 6-membered heterocyclic group, the compound [as one or the group - 0
is a saturated nitrogen-containing 6-membered heterocyclic group, the compound [
1′].

[White] [I″]

[In the formula, R1, F2 and A have the same meanings as before, the group in formula [1'] is an unsaturated nitrogen-containing 6-membered heterocyclic group, and the group in formula [1'] is a saturated In this specification, the formula [l'
] type salts such as pyridiniophosphate, pyrimidiniophosphate, pyridaciniophosphate, and salts of the formula (1') such as piperidinophosphate, piperazinyl phosphate. These are named for convenience, but all of them are included in the salts of the compounds represented by the general formula.

In addition, the salt of the target compound 1) may include the above-mentioned inner salt o.
Also included are intermolecular salts such as those represented by formulas [1a], [Ib], and [IC].

(Ia) (l1l)) [In the formula, R1, R2, A and group -0 each have the same meanings as before, " means an anionic residue of an acid, and Z+ means a cationic residue of a base, respectively." Suitable "acid 7 anion residues" for Y- include, for example, chloride anion, promide anion, and yo! Halide anions such as hydrogen anions, hydroxide anions, sulfate anions such as methanesulfonyloxy-anions, benzenesulfonyloxy-anions, tosyl-oquine anions, sulfonate anions such as trifluoroacetoxy-anions, etc.
Anions, inorganic or organic anionic residues such as carboxylate anions such as tertarroyloxy-anions may be mentioned.

Suitable [base cationic residues] represented by Z+ include:
For example, alkali metal ions such as sodium ions and potassium ions are included. The salt of the target compound (1) includes:
Also included are hydrates and solvates thereof.

The target compound (1) includes all optical isomers based on the asymmetric carbon atom in the molecule of the compound (1).

Furthermore, in this specification, in order to simplify the description, all chemical formulas of the 1,2-propanediol moieties will be represented by formula (a) instead of formula (b).

[In the formula, R has the same meaning as before]

The object compound of this invention and its salt can be produced by the following method.

Manufacturing method (vl) [V]
r group 2 [In the formula, R, RSA and group -0 each have the same meaning as before, R3 means a protecting group for a hydroxy group, and X means an acid residue, respectively] (a) Synthesis of compound ffl: Compound ff) is compound m) or its salt and compound (5)
) in the presence of a base such as dibutyltin dichloride, triethylamine, trimethylamine, pyridine, etc., in an inert solvent such as benzene, toluene, etc., by heating or under heating according to a conventional method. It is obtained by

Suitable [hydroxy protecting group] represented by R5 includes, for example, lower alkoxycarbonyl such as methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, aralkyloxycarbonyl such as benzyloxycarbonyl, alkanoyl such as formylthyl, benzyl, trityl, etc. Commonly used protecting groups such as aralkyl are mentioned.

(1) Synthesis of compound [group] Compound [button] can be produced by subjecting compound [V] or a salt thereof to an elimination reaction of a hydroxy protecting group.

The elimination reaction of the hydroxy protecting group can be carried out by a known method such as reduction or hydrolysis depending on the type of the protecting group.

(c) Synthesis of compound [11] Compound m is the synthesis of compound [button or its salt], compound (IV
It can be prepared by stirring I and an organic solvent at room temperature or under heating. Suitable acid residues represented by X in compound rlV1 include, for example, fluorine, chlorine,
Examples include halogens such as bromine and iodine, and acyloxy groups such as benzenesulfonyloxy and tosyloxy.

It can be produced by reacting the salt with a nitrogen-containing 6-membered heterocyclic compound or a salt thereof.

This reaction is usually carried out in a solvent such as acetone, methanol, tetrahydrofuran, chloroform, or benzene, but any solvent can be used as long as it does not adversely affect the reaction. When the nitrogen-containing 6-membered heterocyclic compound or its salt is a liquid, it can also be used as a solvent.

Although the reaction temperature is not particularly limited, it is desirable to carry out this reaction at room temperature or under heating.

In this reaction, the target compound is of the formula (I
When obtained in the form of intermolecular salts such as those represented by Formula [1'] 1 can be changed into an inner salt represented by (1').

The target compound produced by the above production method can be isolated from the reaction mixture and purified by conventional methods.

The target compound [11] in free form can be converted into its salt form as desired, and the target compound in the form of an internal salt can also be converted into an acid addition salt by a conventional method.

Among the raw material compounds, the new compounds can be produced by the production methods shown in the Examples below or by methods equivalent thereto.

Examples of the salt of the raw material compound include the same salts as mentioned above as the salt of the target compound.

The following pharmacological test data is based on the target compound of this invention [11
has been shown to exhibit high antitumor activity.

Test Method A BALB/C female mice, 12 weeks old and weighing 19 to 212 mm, were used in each group of 6 mice.

Fibrosarcoma Meth A cells (hereinafter abbreviated as Meth A)
used BALB/C Maunus and transplanted and subcultured them in the form of ascites cells every 7 days, and in the test, MethA in the ascites cells collected on the 6th day after transplantation was used as tumor cells.

Each BALB10-v [MethA cell number IX10”
including Q. 0.5 ml Hank's solution was inoculated sarcastically.

Test compounds were dissolved or suspended in 0.5% methylcellulose saline solution and intratumorally injected into each mouse once daily from day 1 to day 4 after implantation at a dose of 100 μV/mouse.

The control group received only vehicle in the same manner.

The antitumor effect of the test compound is expressed by the tumor growth inhibition rate (1-T/C
) was evaluated.

T: Average diameter of tumors in the administration group after 21 days C: Average diameter of tumors in the control group after 21 days Test compound Example 1-(4) and target compound of Example 4 Average - Rice = Tumor growth inhibition rate (capital) ((1-T/C)Xl 0
0) Rice C: Ratio of mice with no tumor growth [number of mice with no tumor growth/number of mice used as a group] Rice d: Average body weight at the time of Meth A transplantation (FA rice e:
Average body weight on day 4 after transplantation (7) Test method B BAL 8-9 weeks old, weight 18.0-22.55'
Eight B/C female mice were used per group.

Fibrosarcoma Meth A cells (hereinafter abbreviated as MethA)
used BALB/C mice, and transplanted and subcultured them in the form of ascites cells every 7 days, and in the test, MethA in the ascites cells collected 6 to 7 days after transplantation was used as tumor cells.

Number of MethA cells in each BALB/C mouse: 5 x 10”
0.1 m+/ of Hank's solution was inoculated intrathoracically.

The test compound was dissolved in a phosphate buffered saline solution and injected intrathoracically into each mouse at a dose of 100 μ910.05 m 1 5 times: 14 days before tumor implantation, 1 hour after tumor implantation, and 3 days after tumor implantation.

The control group received only vehicle in the same manner.

The antitumor activity of the test compounds was evaluated by comparing the mean survival days of the two groups.

T: Average survival days of the drug group C: Average survival days of the control group Test compound Example 2-(4), Example 3-(4), and Example 4
Compound f: T/CXI 00 As is clear from the above test results, the target compound (1) of the present invention exhibits antitumor activity without adversely affecting body weight, and is useful as an antitumor agent.

The active ingredient is usually 0.1" in the form of preparations such as tablets, granules, powders, capsules, syrups, injections, and suppositories.
P/# ~500' l/ky (D administration fi-t' is administered 1 to 4 times a day. However, the above dosage may be increased or decreased depending on the age, body weight, condition of the patient, or administration method.

The various formulations described above can be manufactured by conventional methods using conventional carriers and additives.

The present invention will be explained in more detail below according to examples.

Example 1 Racemic-1-0-octadecy/L'-3-0-)lythylglycerin (9,981) and dibutyltin dichloride (
After adding methyl isocyanate (7,09) to a solution of 52 mg) in dry toluene (150 l),
Stir at 0°C for 3 hours. When the reaction solution was concentrated under reduced pressure, 12
．． 985' racemic-1-〇-octadecytv-2-
0-methylcarbamoy/l/-3-0-)! J Chilluglycerin is obtained as an oil.

IR (film): 3330, 3050.2900, 2
850°1740ff' The obtained compound can be used in the next reaction without purification.

The compound obtained in (1) (15SM methylene chloride (1
00m/) solution, add trifluoroacetic acid (15,7+wf
After adding the ingredients all at once, stir at room temperature for 8 minutes. Ice water (100 g/) is added to the reaction solution, the organic layer is separated, washed with an aqueous sodium bicarbonate solution and water, and then concentrated under reduced pressure. The obtained residue was dissolved in n-hexane (50 g?), filtered, and the filtrate was concentrated under reduced pressure to give a waxy solid racemic-1-0-,octadecyA/-2-0-methylcarbamoyl-glycerin. We get (8,447).

IP (nuji=a/l'): 3380, 1670
cI12-bromoethyl dichlorophosphate (7,6
Add the above (29) to a dry chloroform (jaw?) solution of
A solution of the compound (8,4y) obtained in step 2) and triethylamine (3,181) in dry chloroform (10 t) was added
Add dropwise over 30 minutes at below 0°C. The reaction solution was heated at the same temperature for 2
After stirring for 0 min and then for 1 h at room temperature, it is cooled in a water bath.

Water (10.5 g
/) and pyridine (21 ml) was added dropwise at below 15°C. After the dropwise addition was completed, the mixture was stirred at room temperature for 50 minutes.
The solvent is removed under reduced pressure. The residue is dissolved in a mixture of sodium hydrogen hydroxide aqueous solution and diethyl ether, and the aqueous layer is separated. The aqueous layer was washed with diethyl ether, and purified with 10% hydrochloric acid.
After adjusting to H1, extract twice with ethyl acetate. When the extract was washed with water, dried and concentrated under reduced pressure, 7.42 racemic -1-〇-
Octadecy/l/-2-0-methylcarbamoyl-glycerin-5-(2-prowamoethyl/I/) phosphate is obtained as a beak-like solid.

Engineering'R (nuji v-zshi): 3300.1700.156
0.1210a++ 'A mixture of compound (2,91') obtained in (3) above and pyridi 7 (9 g/) was added to 10
After stirring for 1.5 hours at 0'C, concentrate under reduced pressure. The residue was triturated with acetone, then collected and dissolved in 90% aqueous methanol (2
6g+/) and chloroform (41). After treating this solution with silver acetate (1,67y) for 1 hour, the precipitate is filtered and washed with methanol. The filtrate and washing solution were combined and dried under reduced pressure, and the residue was collected using silica gel (7 oy.

Column chromatography (eluting with a mixed solvent of chloroform, methanol, and water = 65:25:4) yielded 2.01P racemic-i-o-octadecy/l/-
2-0-methylcarbamoyl-glycerin-5-(2-
(1-Pyridinio)ethyl cophosphate is obtained (recrystallized from a mixed solvent of chloroborum and acetone). Melting point 1
95°C (disassembly) Engineering R (Nuji-1-/L/): 3350
, 5250.1710, 1640°235a1 NMR (CD30D) ppm: 0.90 (3H, m)
, 1.06-1.71 (32H, m), 2.71 (3
H,S), 3.28-4.99 (11H,m).

8.05-9.15 (5H, m) Elemental analysis: Calculated value as C3oH55N207P・1.75H2o: C58,28, H9,54, N4.53
Actual measurement M: 05B, 2B, H9,75, N4.5° Example 2 Same as Example 1-(1), racemic-1-0 to octadecyl-3-0-) lythyl-glycerin (8,22y) and phenyl isocyanate (1,83i) in the presence of butyltin dichloride (42) to give racemic-1
-0-Octadecy/l/-2-0-phenylcarbamoyl-5-0-) lytyl-glycerin (10,40y) is obtained as an oil.

IR (Film): 3300.3050.1735.1
700.1600 Example 1-(2), the above (
The compound-0-phenylcarbamoyl-glycerin (4,21) obtained in 1) is obtained as a waxy solid.

IR (Nujichiichiru): 3400, 3250, 1710.
1620as01 (Similar to Example 1-(3), the compound (4,17Si) obtained in the above (2) and 2-bromoethyldichlorophosphate) (2,835') were reacted to form a racemic -1-
〇-Octadecyl-2-0-phenylcarbamoyl-glycerin-3-(2-bromoethyl)phosphate (3
, 1i as a waxy solid.

Engineering R (nuji = r #): 3300.1695.160
0.1245 cM Similarly to Example 1-(4), the above (3
) compound (2,871 and pyridine (8,6m
1) to form racemic-1-o-octadecy/L/
-2-0-phenylcarbamoyl-glycerin-3-C
2-(1-pyridinio)ethyl]phosphate)(1,7
2P) melting point 195°C (decomposition) IR (Nujol): 3300.1725.1600.
1220cm NMR (CD50D) ppm: 0.90 (
3H, m), 1.10-1.71 (32H, m),
3.38-5.00 (11H, m), 6.96-7.4
8 (5H, m), 7.91-8.98 (5H, m)
Elemental analysis: C35H57N207P・5/2 Calculated value as H20: C62,20, H8,95, H4,15
Actual values: C62,50, H8,89, N3.77 Example 5 Similarly to Example 1-(1), racemic-1-o-octadecy/L/-3-0-) lythylglycerin (10, 57
2) Totrichloroacetylisocyanate) (4292)
and racemic-1-〇-octadecyl-2-
0-lupamoyl-5-0-) lythylglycerin (6
, 91y) are obtained. Melting point 75°CIn (Nushor):
3450.3300.1705.1680.1600a
mSimilarly to Example 1-(2), the compound (6,35') obtained in the above (1) was treated with trifluoroacetic acid (6d) to give racemic-1-〇-octadecytL/-2 -0-carbamoyl-glycerin (2,981 is obtained.

Melting point 77°C IFt (Nujo-+): 5420, 1675, 1
62011111M In the same manner as in Example 1-(3), the compound (2,71y-) obtained in the above (2) was reacted with 2-promoethyldichlorophonufate (2,54ti) to produce racemic-1 −
0-octadecyl-2-0-carbamoyl-glycerin-3-(2-bromoethyl/I/)phosphate (1,9
Get 7i. Melting point 76°CIR (nuji a-1v): 33
00.3150.1660.1070°1095 cI
i″1 In the same manner as in Example 1-(4), the compound (1,95F) obtained in the above (3) was reacted with pyridine (9 g/) to produce racemic-1-0-octadecy)v- 2-0-Carbamoyl-glycerin-3-(2-(1-hyricinio)ethyl)phosphate-1-(1,301t-i. Melting point 202
°C (decomposition) 1 1R (nuji rose): 3350.3150.1715.
1630.1235αNMR (CD30D) ppm: 0
．． 89 (3H, t, J=5Hz), 1.04-1.6
4 (32H, m), 3.36-4.94 (11H, m)
, 8.00=9.04 (5H, m) Elemental analysis: C2, H55N207P-H2o calculated value: C58,96, H9,1, N4.74 actual value:
C58,46, H9,61, N4.73 Example 4 In the same manner as Example 1-(4), racemic-1-0-octadecyl-2-〇-ruva obtained in Example 5-<3) Moyl glycerin-5-<2-bromoethyl) phosphate (3,509) and N-methylpiperazine (12,2 g
/) to form racemic-1-〇octadecyl-2
-0-rubamoylglycerin-3-(2-(4-methy/L/-1-piberazini/L/)ethyl)honufate (2,36i is obtained.

Melting point 153°C (decomposition)
5,10551m 'NME (CD30D)pp,:
0.9 (3H, m), 1.04-1.8 (32H,
m), 2.70 (3H, S), :28-3.2 (10H
, m).

3.4-5.7 (4H, rn), 3.9-4.2 (
4H, m).

5.0 (IH, m) Elemental analysis: C29H6oN307P, calculated value as 2H20: C55,30, Hl 0.24. N6.67 actual value: C55,48, H9,91, N6.58 Patent applicant Fujisawa Pharmaceutical Co., Ltd.

Claims (1)

[Claims] (1) General formula: [In the formula, R1 is an alkoxy group; R2 is a hydrogen atom, a lower alkyl group, or an aryl group; A is a lower alkylene group, and the group -0 is substituted with a lower alkyl group. Nitrogen-containing 6
phospholipid derivatives and salts thereof, each representing a membered heterocyclic group. (2) Formula: [In the formula, R1 is an alkoxy group, R2 is a hydrogen atom, a lower alkyl group, or an aryl group; A is a lower alkylene group;
X means an acid residue respectively] or a salt thereof is reacted with a nitrogen-containing 6-membered heterocyclic compound or a salt thereof which may be substituted with a lower alkyl group, and A have the same meanings as above, and group -0 means a nitrogen-containing 6-membered heterocyclic group which may be substituted with a lower alkyl group] or a salt thereof. (6) Formula: [In the formula, R1 is an alkoxy group; R is a hydrogen atom, a lower alkyl group, or an aryl group; A is a lower alkylene group;
O is nitrogen-containing 6 which may be substituted with a lower alkoxy group
An antitumor agent comprising a phospholipid derivative or a salt thereof as an active ingredient.