JPH0687845A - New gamma-lactone derivative - Google Patents

Abstract

PURPOSE:To obtain a new compound useful as an antitumor agent. CONSTITUTION:The gamma-lactone derivative of formula I [R is formula II (X is CH=CH-CH2 or CH(OH)-CH2); n is 1-3); R<1> are R<2> are H or lower alkyl; m is 0-1], e.g. (5R)-4,5-dihydro-5-[(9Z,12Z)-9,12-octadecadienoyloxymethyl]-2(3H)- furanone. This compound of formula I is obtained by reacting a 4,5-dihydro-5- hydroxymethyl-2(3H)-furanone of formula III with 4-dimethylaminopyridine in the presence of 1,3-dicyclohexylcarbodimide and linoleic acid in a solvent such as dichloromethane. The compound of formula I is capable of manifesting excellent synergistic effects by its use with an adriamycin anticancer agent in combination.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel γ-lactone derivative useful as an antitumor agent.

[0002]

2. Description of the Related Art Conventionally, various types of antitumor agents such as plant alkaloid-derived podophyllotoxin (PC) and fungal-derived camptotensin (CT) have been known. Further, the improvement thereof is also active, and derivatives of unsaturated higher fatty acid residues introduced into these PC and CT are also known. More specifically, it has been reported that PC prepared by introducing an unsaturated higher fatty acid ester residue into the 4-position side chain exhibits strong activity, and its antitumor activity is greatly influenced by the stereochemistry at the 4-position. . On the other hand, in the case of CT, C having a polyhydroxylated higher fatty acid ester residue at position 20
It has been reported that T exhibits strong activity.

[0003]

DISCLOSURE OF THE INVENTION We have tried to find a new antitumor agent, focusing on the effect of these higher fatty acid residues on the antitumor activity. As a result, they have found that a novel γ-lactone derivative obtained by introducing a higher fatty acid residue into the ring carbon atom of γ-lactone has the desired antitumor activity, and completed the present invention. This γ-lactone derivative has antitumor activity by itself,
It exhibits an excellent synergistic effect when used in combination with an adriamycin anticancer agent.

[0004]

That is, according to the present invention,
The novel γ-lactone derivative is represented by the following general formula [I].

[0005]

[Chemical 1]

(Where R is

[0007]

[Chemical 2]

Be [0008]; X is, -CH = CH-CH ₂ - or -CH (OH) -CH (OH) -CH 2 - and is;
R ¹ and R ² may be the same or different and each is a hydrogen atom or a lower alkyl group; n is an integer of 1 to 3; m is an integer of 0 to 1. ) Specific examples of -CO-R include unsaturated higher fatty acid residues such as oleic acid, linoleic acid, and linolenic acid as described below, or polyhydroxylated products thereof.

[0009]

[Chemical 3]

Next, one example of the method for producing a γ-lactone derivative according to the present invention will be described in detail in the case where R is a linoleic acid residue (R ^a ) or its polyhydroxylated product (R ^b ) according to the following scheme. explain. Of course, even when R is oleic acid, linolenic acid, or the like, the target compound can be similarly prepared.

[0011]

[Chemical 4]

1. 4,5-Dihydro-5-hydroxymethyl-2 (3H) -furanone (Compound 1) and 4,5
-Dihydro-3-hydroxy-4,4-dimethyl-2
Production of unsaturated higher fatty acid esters (compound 2 and compound 5) of (3H) -furanone (compound 4); basically an esterification reaction of an acid and an alcohol, and an ester that can be generally used for this type of reaction Any known method may be adopted as long as it is a chemical reaction. For example, the ester compound 2 is an organic solvent solution of 4,5-dihydro-5-hydroxymethyl-2 (3H) -furanone (Compound 1) and 4-dimethylaminopyridine (DMAP) under an inert gas atmosphere, for example, It can be obtained by adding an organic solvent solution obtained by dissolving linoleic acid and 1,3-dicyclohexylcarbodiimide (DCC) to a dichloromethane solution under ice cooling, for example, a dichloromethane solution and stirring the mixture for several tens of minutes to several hours. The ester compound 5 can also be obtained by the same method as described above. Instead of linoleic acid, the acid chloride may be used for esterification by a known method.

2. Production of polyhydroxy compound 3 and compound 6 by oxidation of compound 2 and compound 5; compound 3 and compound 6 are obtained by subjecting unsaturated higher fatty acid ester compound 2 and compound 5 obtained above to a known appropriate oxidation reaction. It is obtained by attaching. A preferred oxidation reaction is
This is a so-called osmic acid oxidation reaction by the combined use of N-methylmorpholine N-oxide and OsO ₄ . That is, the target compound, polyhydroxy compound 3, is obtained by adding N-methylmorpholine N-oxide to an acetone / water mixed solution of unsaturated higher fatty acid ester compound 2 and further adding OsO ₄ and stirring the mixture at room temperature for several days. Obtained by The compound 6 can also be obtained by subjecting the compound 5 to the same operation as above. The method for producing these compounds 2, 3, 5, 6 is not limited to the above method.

The compounds of the present invention are shown in Table 1 below.
It is as shown in Table-2. The compound of the present invention has a chiral center at a cyclic carbon atom, and any of the R isomer, the S isomer, or a mixture thereof is included in the scope of the present invention.

[0015]

[Table 1]

[Table 2]

The drug containing the compound of the present invention as an active ingredient is
By a conventional method, the compound is added as a pharmaceutical aid, for example, a pharmaceutically acceptable carrier, diluent, excipient, binder, disintegrating agent, lubricant, preservative, stabilizer, solubilizing agent, Obtained by formulating into a unit dosage form suitable for administration, such as tablets, capsules, powders, granules, microcapsules, syrups, elixirs, injections, suppositories, etc., together with flavoring agents and the like. be able to. Tablets, capsules, powders, solid preparations such as granules, the compound of the present invention, lactose, mannitol, glucose, hydroxypropylcellulose, microcrystalline cellulose, carboxymethylcellulose, starch, polyvinylpyrrolidone, aluminum metasilicate, Carriers or diluents such as talc; Lubricants such as magnesium stearate; Disintegrators such as calcium fibrin gluconate; Dissolution aids such as glutamic acid, aspartic acid; Stabilizers such as lactose be able to. If necessary, tablets may be coated with a gastric or enteric substance such as sucrose, gelatin, hydroxypropylcellulose, hydroxypropylmethylcellulose, or soft capsules. Syrup, elixir, solution,
In the case of liquid preparations such as emulsions and suspensions, the compound of the present invention is dissolved or dispersed in a pharmaceutically acceptable liquid medium such as purified water, physiological saline, buffer solution, ethanol and the like. Further, if necessary, a surfactant, a sweetener, a flavor, a fragrance, an antiseptic and the like can be appropriately added to prepare a formulation. On the other hand, injections for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions and emulsions. Such injections are prepared by diluting the compound of the present invention with water for injection, an aqueous diluent such as physiological saline or polyethylene glycol, propylene glycol, olive oil,
It can be prepared by mixing with a water-insoluble diluent such as ethanol. In addition, if necessary,
Auxiliary agents such as preservatives, wetting agents, surfactants, dispersants, stabilizers and solubilizing agents can also be included. These injections can usually be sterilized by filtration using a bacteria-retaining filter or the like, blending of a bactericide, irradiation, etc., and after further treatment, a solid preparation is obtained by a method such as freeze-drying. Immediately before use, sterile water or a sterile diluent for injection can be added for use. The compound of the present invention can be administered orally or rectally, or parenterally, such as intravenously, intramuscularly and subcutaneously. The dose may be varied depending on the type of compound used, the method of administration, the severity of the symptoms of the patient to be treated, the age and weight of the patient, the judgment of the doctor, etc. 1 to about 500 mg / Kg body weight once a day or 2 to 4
It is appropriate to administer in divided doses. However, the above dose range is merely a guide, and it is of course possible to administer an amount above or below the above range depending on the judgment of the doctor, the severity of symptoms, and the like.

Next, the following tests were conducted on the antitumor activity of the novel γ-lactone derivative according to the present invention, the synergistic effect in combination with adriamycin, and the vascular endothelial cell adhesion inhibitory activity. (1) In vitro antitumor test (i) Each test substance was dissolved in DMSO and RPMI16
It was diluted with 40 medium + 10% FCS + 10 μM 2-mercaptoethanol to a final concentration of 0.001 to 100 μg / ml. (Ii) P388AD in the logarithmic growth phase in the above test solution
M ^r cells at 3 × 10 ³ cells / well, total flow rate 10
After adding 0 μl and culturing for 48 hours, the number of viable cells was measured by the MTT method to determine the IC ₅₀ value. The results are shown in the table below.

[0018]

[Table 3]

(2) Synergistic effect of the combined use with adriamycin (i) The test substance (Compound No. 1, Example 2) was added to DMS.
Dissolve in O, RPMI1640 medium + 10% FCS + 1
0 μM 2-mercaptoethanol to give a final concentration of 0.00
It was diluted to 1 to 100 μg / ml. (Ii) Adriamycin was added to the above test solution to a final concentration of 0.01 to 10 μg / ml, and P388ADM ^r cells in the logarithmic growth phase were further added to 3 × 10 ³ c.
wells / well at a total flow rate of 100 μl, and after culturing for 48 hours, the number of viable cells was measured by the MTT method. (Iii) The concentration of the test substance that inhibits the proliferation to half of the number of cells in the target well containing no test substance and adriamycin was defined as the IC ₅₀ value. (Iv) When the test substance is present at a certain concentration, IC ₅₀
The concentration of adriamycin showing the inhibition of cell growth equal to the value was calculated, and GADDUM prot was performed to determine the synergistic effect of the test substance. The results are as shown in FIG. According to this, the original IC _{50 of the} test substance is 18.7 μg / ml and that of adriamycin is 1.5 μg / ml, but when considered by the arithmetic mean, the concentration of the test substance is 5.0 μg / ml. , The required concentration of adriamycin to achieve the same effect, ie to achieve the same IC ₅₀ , was 1.08.
According to the above test results, 0.56 μg / ml actually shows the same level of action and effect as μg / ml. Therefore, it can be said that the action of adriamycin was enhanced by about 1.9 times by the combined use of 5.0 μg / ml of the test substance.

[0020]

EFFECTS OF THE INVENTION According to the above test results, the γ-lactone derivative of the present invention has an antitumor activity by itself, and exhibits a remarkable synergistic effect in combination with an adriamycin anticancer agent, and is useful as an antitumor agent. is there.

[0021]

[Examples] Examples will be described in detail below, and the meanings of the respective abbreviations are as follows. THF: Tetrahydrofuran NMO: N-Methylmorpholine N-oxide IBCF: Isobutyl chloroformate Pd-C: Palladium-carbon DMAP: 4-Dimethylaminopyridine DCC: 1,3-dicyclohexylcarbodiimide Further, the present invention is not limited to these examples. It is not limited. (Example 1) (5R) -4,5-dihydro-5-[(9Z, 12Z)
-9,12-Octadecadienoyloxymethyl] -2
Production of (3H) -furanone [(R) -2]; (5R) -4,5-dihydro-5- (hydroxymethyl) -2 (3H) -furanone [(R) -1] (under a nitrogen atmosphere. 750
mg, 6.7 mmol) and 4-dimethylaminopyridine (DMAP) (50 mg, 0.41 mmol) in a dichloromethane (2 ml) solution, under ice cooling, with linoleic acid (2.
01 ml, 6.5 mmol) and 1,3-dicyclohexylcarbodiimide (DCC) (1.09 g, 6.5 m)
Dichloromethane (2 ml) solution was added, and the mixture was stirred for 3 hours. The generated DC urea was filtered off, washed with a 5% HCl aqueous solution, a saturated sodium hydrogen carbonate aqueous solution and a saturated saline solution, and dried with magnesium sulfate. The residue obtained by concentration is purified by silica gel column chromatography (dichloromethane) to give a colorless transparent oily substance [(R)-
2] (2.40 g, 98%) was obtained. [Α] ²⁴ _D : -17.69 ° (c = 1.41, CHCl ₃ ) IR (film) cm ^-1 : 3009,2931,2856,2360,1785,1741,1462,1
348,1158,1076,948,725 ¹ H-NMR (200MHz, CDCl ₃ ): 0.89 (t, 3H, J = 6.7Hz), 1.20-1.46
(m, 14H), 1.51-1.72 (m, 2H), 1.94-2.14 (m, 4H), 2.27-2.77
(m, 3H), 2.51-2.65 (m, 2H), 2.77 (t, 2H, J = 5.9Hz), 4.15 (dd,
1H, J = 12.2,5.4Hz), 4.31 (dd, 1H, J = 12.2,3.4Hz), 4.70-4.7
8 (m, 1H), 5.21-5.35 (m, 4H) HRMS (C ₂₃ H ₃₈ O ₄ ): Calculated 378.2770 Found 378.2788 (M ⁺ ).

(Example 2) (5S) -4,5-dihydro-5-[(9Z, 12Z)
-9,12-Octadecadienoyloxymethyl] -2
Production of (3H) -furanone [(S) -2]; (5S)-
4,5-dihydro-5- (hydroxymethyl) -2 (3
Example 1 except that H) -furanone [(S) -1] was used.
The title compound [(S) -2] was obtained in the same manner as in (Yield 88%). [Α] ²⁴ _D : + 18.36 ° (c = 1.13, CHCl ₃ ) IR (film) cm ^-1 : 3009,2928,2856,1785,1741,1462,1348,1
158,1076,948,726 ¹ H-NMR (200MHz, CDCl ₃ ): 0.89 (t, 3H, J = 6.7Hz), 1.20-1.46
(m, 14H), 1.51-1.72 (m, 2H), 1.94-2.14 (m, 5H), 2.27-2.77
(m, 3H), 2.51-2.65 (m, 2H), 2.77 (t, 2H, J = 5.9Hz), 4.15 (dd,
1H, J = 12.2,5.4Hz), 4.32 (dd, 1H, J = 12.2,3.4Hz), 4.70-4.7
8 (m, 1H), 5.21-5.35 (m, 4H) HRMS (C ₂₃ H ₃₈ O ₄ ): Calculated 378.2770 Measured 378.2774 (M ⁺ )

Example 3 (3R) -4,5-dihydro-4,4-dimethyl-3-
Preparation of [(9Z, 12Z) -9,12-octadecadienoyloxy] -2 (3H) -furanone [5]; (3
R) -4,5-Dihydro-3-hydroxy-4,4-dimethyl-2 (3H) -furanone (4) was used, but the procedure was the same as in Example 1 to obtain the title compound (5). (89
%) Was obtained. [Α] ²⁴ _D : -5.46 ° (c = 2.01, CHCl ₃ ) IR (film) cm ^-1 : 3009,2927,2856,1796,1752,1466,1400,1
377,1152,1102,1033,1015,999,723 ¹ H-NMR (200MHz, CDCl ₃ ): 0.89 (t, 3H, J = 6.6Hz), 1,11 (s, 3
H), 1.21 (s, 3H), 1.20-1.50 (m, 14H), 1.51-1.76 (m, 2H), 1.9
4-2.17 (m, 4H), 2.41-2.50 (m, 2H), 2.771 (t, 2H, J = 5.9Hz),
4.01 & 4.06 (ABd, 2H, J = 10.5Hz), 5.21-5.35 (m, 5H) HRMS (C ₂₄ H ₄₀ O ₄ ): Calculated 392.2925 Measured 392.2923 (M ⁺ )

(Example 4) (5R) -4,5-dihydro-5- (9,10,12,
Production of 13-tetrahydroxyoctadecanoyloxymethyl) -2 (3H) -furanone [(R) -3]; Compound [(R) -2] (2.86 g, 7.5 mmol) in acetone-water (9 1) (100 ml) solution in N-methylmorpholine N-oxide (NMO) (2.65 g,
22.6 mmol) and further OsO ₄ (4% aqueous solution 2.40 ml, 0.38 mmol) was added, and the mixture was stirred at room temperature for 5 minutes.
It was stirred for a day. Aqueous sodium hydrogen sulfate solution (5 ml) was added and the mixture was stirred for 20 minutes, and then acetone was distilled off under reduced pressure.
The residue was extracted with acetic acid methyl ester, washed with saturated brine, dried over magnesium sulfate and concentrated, and the residue obtained was recrystallized from methanol to give the title compound as a white powder [(R)-
3] (2.80 g, 82%) was obtained. mp: 158-159 ℃ IR (KBr) cm ^-1 : 3308,2931,2857,1779,1736,1469,1380,11
69,1065,984,949,857,807,724 ¹ H-NMR (200MHz, CDCl ₃ ): 0.91 (t, 3H, J = 5.9Hz), 1.15-1.72
(m, 22H), 1.85-2.43 (m, 4H), 2.52-2.64 (m, 2H), 3.36-3.75
(m, 4H), 4.16 (dd, 1H, J = 12.2,5.6Hz), 4.32 (dd, 1H, J = 12.2,
2.9Hz), 4.72-4.96 (m, 1H) HRMS (C ₂₃ H ₄₃ O ₈ ) M + 1: Calculated value 447.2956 Measured value 447.2937 (M + 1)

(Example 5) (5S) -4,5-dihydro-5- (9,10,12,
Preparation of 13-tetrahydroxyoctadecanoyloxymethyl) -2 (3H) -furanone [(S) -3]; the same operation as in Example 4 except that the compound [(S) -2] was used. The title compound [(S) -3] (84%) was obtained. mp: 155 ℃ IR (KBr) cm ^-1 : 3294,2931,2854,1779,1735,1741,1381,11
69,1065,984,950,919,881,857,807,725 ¹ H-NMR (200MHz, CDCl ₃ ): 0.91 (t, 3H, J = 5.9Hz), 1.15-1.72
(m, 22H), 1.85-2.43 (m, 4H), 2.52-2.64 (m, 2H), 3.36-3.75
(m, 4H), 4.16 (dd, 1H, J = 12.2,5.5Hz), 4.31 (dd, 1H, J = 12.3,
3.1Hz), 4.72-4.96 (m, 1H) HRMS (C ₂₃ H ₄₃ O ₈ ) M + 1: Calculated value 447.2957 Measured value 447.2956 (M + 1)

(Example 6) (3R) -4,5-dihydro-4,4-dimethyl-3-
Production of (9,10,12,13-tetrahydroxyoctadecanoyloxy) -2 (3H) -furanone (6);
The title compound (6) was obtained by the same procedure as in Example 4 except that the compound (5) was used (yield 85%). mp: 148-155 ℃ IR (KBr) cm ^-1 : 3753,3241,2917,2855,1794,1747,1470,13
77,1299,1160,1052,919,859,723 ¹ H-NMR (200MHz, CDCl ₃ ): 0.91 (t, 3H, J = 6.5Hz), 1.08 (s, 3
H), 1.12 (s, 3H), 1.21-1.74 (m, 22H), 2.47 (t, 2H, J = 7.3Hz),
3.36-3.75 (m, 4H), 4.00-4.16 (m, 2H), 5.51 (s, 1H) HRMS (C ₂₄ H ₄₅ O ₈ ) M + 1: Calculated 461.3114 Measured 461.3120 (M + 1)

[0027]

[Brief description of drawings]

FIG. 1 is a diagram showing a synergistic effect of the combined use with adriamycin.

Claims (2)

[Claims] 1. A γ-lactone derivative represented by the following general formula [I]. [Chemical 1] Where R is By and; X is, -CH = CH-CH ₂ - or -CH (O
H) —CH (OH) —CH ₂ —; R ¹ and R ² may be the same or different and each is a hydrogen atom or a lower alkyl group; n is an integer of 1 to 3; , And an integer of 0 to 1. 2. The γ-lactone derivative according to claim 1, wherein n is 2.