JP2640245B2 - 1,4-dihydropyridine derivative - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to a novel 1,4-dihydropyridine derivative having excellent pharmacological activity, and more specifically, a 1,4-dihydropyridine derivative useful as a therapeutic agent for tumors and the like. About.

[Prior art] Many compounds have been already known for 1,4-dihydropyridine derivatives.

Among the known 1,4-dihydropyridine derivatives, there are many compounds known to have pharmacological activity,
Most of them have pharmacological activity on the circulatory system, and other pharmacological activities have anti-inflammatory effects,
Only a few have reported hepatoprotective effects.

On the other hand, 1,4-
For the dihydropyridine derivative, see Japanese Patent Publication 55-500577.
The publication describes that a 1,4-dihydropyridine compound having no substituent at the 4-position has a transfer inhibiting effect on certain tumors. JP-A-60-6613 describes an antitumor and antitumor metastatic agent containing 1,4-dihydropyridine such as nifedipine or nimodipine as an active ingredient. Further, Japanese Patent Application Laid-Open No. 62-87516 describes a method for treating a malignant tumor using a platinum coordination compound in combination with a compound such as nifedibin or nimodibin.

[Problems to be solved by the present invention] However, Japanese Patent Application Laid-Open Nos.
The invention described in 87516 discloses that a drug having a calcium pathway-blocking effect is used as an antitumor drug or used in combination with a platinum coordination compound that is an antitumor drug, and is not necessarily practical in terms of side effects. There is a disadvantage that. In other words, calcium pathway blockers are potent drugs from the beginning, and they act on the heart, blood vessels, etc., even in very small amounts.
It has the disadvantage that adverse effects on the vasculature are inevitable.

The present inventors have extensively screened the 1,4-dihydropyridine derivative for the combined effect with an antitumor drug and the presence or absence of a calcium pathway blocking effect. In particular, they have found that they have the effect of significantly increasing the sensitivity of tumor cells that have acquired resistance and do not have the calcium pathway blocking effect, and have reached the present invention.

[Means for Solving the Problems] According to the present invention, formula (I) (In the formula, R represents a pyridyl group, an N-morpholyl group, or an N-benzyl-N-methylamino group which may be substituted with one methyl group or ethyl group, and n represents an integer of 1 to 4. ) Is provided.

In the present invention, among the 1,4-dihydropyridine derivatives represented by the above formula (I), those having a particularly remarkable action of enhancing drug sensitivity are those wherein R is a pyridyl group or a pyridyl group substituted by one methyl group. , N is an integer of 1 to 3.

As described in detail in later test examples, the compounds having these specific substituents have a more remarkable drug sensitivity-enhancing effect than other compounds, and are particularly useful as medicaments.

All of the 1,4-dihydropyridine derivatives represented by the above formula (I) can be produced by using a well-known reaction conventionally used for producing 1,4-dihydropyridines. For example, 2-formyl-p-dioxene is reacted with β-aminocrotonate and acetoacetate by heating or heating to reflux in the presence or absence of an organic solvent (method A); It is produced by reacting p-dioxene with acetoacetate and aqueous ammonia in the presence or absence of an organic solvent by heating, preferably by heating to reflux (method B). These reactions are represented by the following reaction formulas.

Method A Method B The reactions used in these production methods are conventionally 1,4
-Known reactions used in the production of dihydropyridine compounds (for example, used in the methods described in JP-B-46-40625, JP-B-56-37225, JP-A-60-214786, etc.) Reaction). Therefore, the 1,4-dihydropyridine derivative of the present invention can be produced by appropriately applying other reactions described in these known documents in addition to the above-mentioned method.

The compounds of the family family used in the above production method are all known compounds and can be easily obtained or produced by those skilled in the art as needed. That is,
Both acetoacetate and β-aminocrotonate are compounds that are commonly used as raw materials for the production of 1,4-dihydropyridine compounds, and commercially available products can be obtained as needed and can be easily synthesized. be able to. Also, 2-formyl-p-dioxene is MSSh
ostakovskii; Izvest.Akad.Nauk.SSSROtdel.Khim.Na
uk., 1685, (1961).

According to the present invention, the reaction product produced by the above-mentioned method, that is, the 1,4-dihydropyridine derivative represented by the formula (I) is prepared by a conventional method such as extraction with a solvent, chromatography, crystallization and the like. From and purified.

[Examples] Next, examples will be described to describe synthesis examples of the 1,4-dihydropyridine derivative according to the present invention and results of pharmacological tests performed to confirm its usefulness. It goes without saying that the present invention is not limited to the embodiments.

Example 1 4- [2- (5,6-dihydro-p-dioxinyl)]-
Synthesis of 2,6-dimethyl-1,4-dihydropyridine-3,5-dicarboxylic acid bis (2-pyridylmethyl) ester 2-formyl-p-dioxene 4.06 g (0.040 mol),
16.4 g of acetoacetic acid-2-pyridylmethyl ester (0.085
Mol) and 6.6 ml of 28% aqueous ammonia were dissolved in 40 ml of isopropyl alcohol, and the mixture was heated under reflux for 20 hours. After cooling, the reaction solution was concentrated, the residue was dissolved in ethyl acetate, and impurities were extracted and removed with a small amount of water. After the ethyl acetate layer was dried over sodium sulfate, the solvent was distilled off and the residue was washed with ethyl alcohol and n-
Crystallization was performed with a mixed solvent of hexane to obtain 6.4 g of crystals. Then, recrystallization from isopropyl alcohol gave 5.48 g (yield 29.3%) of the target substance as white crystals. The analytical values of this substance are as follows.

Melting point: 157.5-160.0 ° C 3300 (NH), 1680 (C = O), 1200 (C-O) NMR (CDCl 3, TMS, PPM) 2.30 (6H, s, 2,6 -position _{CH 3) 3.90 (4H, s} , dioxene ring OCH _{2 CH 2 O) 4.68 (1H,} s, 4 -position H) 5.31 (4H, s, 2 × COOCH 2) 5.85 (1H, s, vinyl H) 6.30 (1H, b, NH) 7.0-7.8,8.4-8.7 ( Elemental analysis (C ₂₅ H ₂₅ N ₃ O ₆ ) Calculated: C, 64.79: H, 5.44: N, 9.07 Found: C, 64.66: H, 5.62: N, 8.86 Performed Example 2 4- [2- (5,6-dihydro-p-dioxinyl)]-
Synthesis of 2,6-dimethyl-1,4-dihydropyridine-3,5-dicarboxylic acid bis [2- (6-methylpyridyl) methyl] ester 2-formyl-p-dioxene 4.00 g (0.035 mol),
16.8 g (0.081 mol) of acetoacetic acid-2- (6-methylpyridyl) methyl ester and 15 ml of 28% aqueous ammonia were dissolved in 25 ml of isopropyl alcohol, and the mixture was heated under reflux for 20 hours. After cooling, the reaction solution was concentrated, the residue was dissolved in ethyl acetate, and impurities were extracted and removed with a small amount of water. After drying the ethyl acetate layer with sodium sulfate, the solvent was distilled off and the residue was subjected to silica gel column chromatography with an ethyl acetate solvent,
The obtained crystals were recrystallized from isopropyl alcohol to obtain 1.70 g (yield 10.0%) of the target substance as white crystals. The analytical values of this substance are as follows.

Melting point: 158.0-160.0 ° C 3270 (NH), 1690 (C =), 1260 (CO) NMR (CDCl ₃ , TMS, PPM) 2.33 (6H, s, 2, 6-position CH ₃ ) 2.53 (6H, s, 2 × pyridine ring 6) position _{CH 3) 3.81 (4H, s} , dioxene ring _{OCH 2 CH 2 O) 4.64 (} 1H, s, 4 -position H) 5.23 (4H, ABq, 2 × COOCH 2) 5.83 (1H, s, vinyl H) 5.86 ( 1H, b, NH) 7.02 (2H, b, 2 × pyridine ring 5-position H) 7.08 (2H, b, 2 × pyridine ring 3-position H) 7.45 (2H, t, 2 × pyridine ring 4-position H) 34- [2- (5,6-dihydro-p-dioxinyl)]-
Synthesis of 2,6-dimethyl-1,4-dihydropyridine-3,5-dicarboxylic acid bis [2- (N-morpholyl) ethyl] ester 2-formyl-p-dioxene 4.00 g (0.035 mol),
Acetoacetic acid-2- (N-morpholyl) ethyl ester 1
7.4 g (0.081 mol) and 15 ml of 28% aqueous ammonia were dissolved in 25 ml of isopropyl alcohol, and the mixture was heated under reflux for 20 hours. After cooling, the reaction solution was concentrated, the residue was dissolved in ethyl acetate, and impurities were extracted and removed with a small amount of water. After the ethyl acetate layer was dried over sodium sulfate, the solvent was distilled off, and the resulting crystals were recrystallized from isopropyl alcohol to obtain 1.30 g (7.3% yield) of the target substance as pale yellow crystals. The analytical values of this substance are as follows.

Melting point: 113.5-115.0 ° C 3320 (NH), 1690 (C = O), 1260 (C-O) NMR (CDCl 3, TMS, PPM) 2.31 (6H, s, 2,6 -position _{CH 3) 2.39-2.79 (12H, m} , 2 × COOCH ₂ CH ₂ , 2 × morpholine ring 3,5
CH ₂ ) 3.53-3.82 (8H, m, 2 × CH _{2 at the} 2,6-morpholine ring) 3.87 (4H, s, dioxene ring OCH ₂ CH ₂ O) 4.06-4.37 (4H, m, 2 × COOCH ₂ CH ₂ ) 4.42 (1H, s, 4-position H) 5.65 (1H, b, NH) 5.88 (1H, s, vinyl H) Example 4 4- [2- (5,6-dihydro-p-dioxinyl)]-
Synthesis of 2,6-dimethyl-1,4-dihydropyridine-3,5-dicarboxylic acid bis [3- (3-pyridyl) propyl] ester 2-formyl-p-dioxene 4.00 g (0.035 mol),
Acetoacetic acid-3- (3-pyridyl) propyl ester 1
7.9 g (0.081 mol) and 15 ml of 28% aqueous ammonia were dissolved in 25 ml of isopropyl alcohol, and the mixture was heated under reflux for 20 hours. After cooling, the reaction solution was concentrated, the residue was dissolved in ethyl acetate, and impurities were extracted and removed with a small amount of water. After the ethyl acetate layer was dried over sodium sulfate, the solvent was distilled off and the residue was subjected to silica gel column chromatography with a mixed solvent of chloroform and methanol, and the obtained crystals were washed with ether to give 2.40 g of the target substance as pale yellow crystals (yield Rate 13.2%). The analytical values of this substance are as follows.

Melting point: 113.5-115.0 ° C 3250 (NH), 1680 (C = O), 1260 (CO) NMR (CDCl ₃ , TMS, PPM) 1.79-2.18 (4H, m, 2 × COOCH ₂ CH ₂ CH ₂ ) 2.32 (6H, s, 2,6 position _{CH 3) 2.74 (4H, t} , 2 × COOCH 2 CH 2 CH 2) 3.92 (4H, s, dioxene ring _{OCH 2 CH 2 O) 3.93-4.41 (} 4H, m, 2 × COOCH 2 CH 2 _{CH 2) 4.52 (1H, s} , 4 -position H) 5.87 (1H, s, vinyl H) 5.96 (1H, b, NH) 7.03-7.27 (2H, m, 2 × pyridine ring 5-position H) 7.35-7.56 ( 2H, m, 2 × pyridine ring 4-position H) 8.26-8.51 (4H, m, 2 × pyridine ring 2,6-position H) Example 5 4- [2- (5,6-dihydro-p-dioxinyl)] −
Synthesis of 2,6-dimethyl-1,4-dihydropyridine-3,5-dicarboxylic acid bis 2- (N-benzyl-N-methylamino) ethyl ester hydrochloride 2.40 g (0.030 mol) of 2-formyl-p-dioxene ,
Acetoacetic acid-2- (N-benzyl-N-methylamino)
Dissolve 14.5 g (0.058 mol) of ethyl ester and 7.0 ml of 28% aqueous ammonia in 50 ml of isopropyl alcohol,
The mixture was heated under reflux for an hour. After cooling, the reaction solution was concentrated, the residue was dissolved in ethyl acetate, the residue was subjected to silica gel column chromatography with a mixed solvent of chloroform and methanol, the resulting oil was dissolved in ethanol, and 2 equivalents of hydrochloric acid were added. The solvent was distilled off. The obtained residue is triturated with ethyl acetate to give 1.82 g of the powdered target substance (9.7% yield).
I got The analytical values of this substance are as follows.

3250 (NH), 1710 (C = O), 1240 (C-O) NMR ( free _{base) (CDCl 3, TMS, PPM} ) 2.24 (6H, s, 2,6 -position _{CH 3) 2.28 (6H, s} , 2 × N-CH ₃ ) 2.68 (4H, t, 2 × COOCH ₂ CH ₂ N) 3.53 (4H, s, 2 × N− ₂ CH ₂ Ph) 3.85 (4H, s, dioxene ring OCH ₂ CH ₂ O) 4.23 (4H, t, 2 × COOCH ₂ CH ₂ N) 4.47 (1H, s, 4-position H) 5.57 (1H, b, NH) 5.85 (1H, s, vinyl H) 7.23 (10H, s, 2 × Ph) Example 6 4- [2- (5,6-dihydro-p-dioxinyl)]-
Synthesis of 2,6-dimethyl-1,4-dihydropyridine-3,5-dicarboxylic acid bis [2- (2-pyridyl) ethyl] ester 2-formyl-p-dioxene 4.00 g (0.035 mol),
Acetoacetic acid-2- (2-pyridyl) ethyl ester 17.0
g (0.082 mol) and 15 ml of 28% aqueous ammonia were dissolved in 100 ml of isopropyl alcohol, and the mixture was heated under reflux for 48 hours. After cooling, the reaction solution was concentrated, the residue was dissolved in ethyl acetate, and impurities were extracted and removed with a small amount of water. After the ethyl acetate layer was dried over sodium sulfate, the solvent was distilled off and the residue was subjected to silica gel column chromatography with a mixed solvent of ethyl acetate and acetone, and the obtained crystals were recrystallized from isopropyl alcohol to give pale yellow crystals. 4.34 g of target substance (yield 25.2
%). The analytical values of this substance are as follows.

Melting point: 130.0-131.0 ° C 3340 (NH), 1690 (C = O), 1210 (C-O) NMR (CDCl 3, TMS, PPM) 2.20 (6H, s, 2,6 -position _{CH 3) 3.13 (4H, t} , 2 × COOCH 2 _{CH 2 CH 2) 3.83 (4H} , s, dioxene ring _{OCH 2 CH 2 O) 4.31 (} 1H, s, 4 -position H) 4.31-4.69 (4H, m, 2 × COOCH 2 CH 2) 5.54 (1H, s, Vinyl H) 5.60 (1H, b, NH) 7.00-7.23 (4H, m, 2 × pyridine ring 3,5-position H) 7.43-7.67 (2H, m, 2 × pyridine ring 4-position H) 8.48 (2H, d Example 7 4- [2- (5,6-dihydro-p-dioxinyl)]-
Synthesis of 2,6-dimethyl-1,4-dihydropyridine-3,5-dicarboxylic acid bis [2- (5-ethylpyridyl) methyl] ester 2-formyl-p-dioxene 5.00 g (0.044 mol),
22.7 g (0.103 mol) of acetoacetic acid-2- (5-ethylpyridyl) methyl ester and 10 ml of 28% aqueous ammonia were dissolved in 30 ml of isopropyl alcohol, and the mixture was heated under reflux for 20 hours. After cooling, the reaction solution was concentrated, the residue was dissolved in ethyl acetate, and impurities were extracted and removed with a small amount of water. After the ethyl acetate layer was dried over sodium sulfate, the solvent was distilled off, and the residue was subjected to silica gel column chromatography with a mixed solvent of n-hexane and ethyl acetate.The obtained crystals were recrystallized from isopropyl alcohol and ether to give a pale yellow color Crystal target substance 4.
90 g (22.1% yield) was obtained. The analytical values of this substance are as follows.

Melting point: 124.0-126.0 ° C 3190 (NH), 1690 (C = O), 1260 (CO) NMR (CDCl ₃ , TMS, PPM) 1.23 (6H, t, 2 × CH ₂ CH ₃ ) 2.31 (6H, s, 2,6 position) _{CH 3) 2.61 (4H, q} , 2 × CH 2 CH 3) 3.89 (4H, s, dioxene ring _{OCH 2 CH 2 O) 4.62 (} 1H, s, 4 -position H) 5.24 (4H, ABq, 2 × COOCH 2 5.86 (1H, s, vinyl H) 5.87 (1H, b, NH) 7.29-7.51 (4H, m, 2 × pyridine ring 3,4 position H) 8.32-8.35 (2H, m, 2 × pyridine ring 6 position H) example 8 (test example) 10 ⁶ vincristine in CDF ₁ mice per group six anticancer agent enhancing effect in vincristine-resistant tumor-bearing mice (VC
R) Implantation of resistant mouse leukemia (P388 / VCR) cells intraperitoneally, administration of the compound of the present invention and VCR intraperitoneally for 5 days,
Observation was performed to determine the number of surviving days, and the survival rate (T / C)% relative to the control was determined. The potentiating effect (T / V)% of the anticancer drug was determined by the following equation. Verapamil (Ve
rapamil). The results are shown in Tables 1 to 5. In the table, Compound 1 is Example 1, Compound 2 is Example 2, Compound 3 is Example 3, Compound 4 is Example 4, and Compound 5 is Compound 5.
Is the compound obtained in Example 5, Compound 6 is the compound obtained in Example 6, and Compound 7 is the compound obtained in Example 7.

Example 9 (Test example) Inhibitory effect on potassium contracture in rat rectal smooth muscle A rat rectum (about 1.5 cm) was excised and suspended in a Magnus tube. Intestinal constriction was induced by using a lock solution as a nutrient solution and changing the potassium (K) concentration of the solution from 5.6 mmol (mM) to 56.0 mM. This contraction is a contraction through a membrane-dependent Ca-channel based on depolarization by K concentration. The contraction force is FD-transducer,
Recorded via polygraph. The compound of the present invention acted 5 minutes before changing the K concentration, and acted simultaneously with changing the K concentration. The concentration of the compound of the present invention in the nutrient solution is
^{They were} 10 ^-8 and 10 ^-7 mol (M). The determination was made 5 minutes after the K concentration change, and the difference in contractile force from that before the K concentration change was indicated by a% suppression value. Verapamil and nicardipine were used as positive control compounds. The results are shown in Table 6. In the table, compound 1 to compound 6 have the same meaning as described above (Example 8).

[Effect of the Invention] The 1,4-dihydropyridine derivative according to the present invention enhances its action when used in combination with an antitumor agent. The effect is particularly significant for clones that have acquired resistance to antineoplastic agents. For example, as is clear from Tables 1 to 5, the antitumor agent alone hardly prolongs the life of the antitumor agent alone when the P388 / VCR cells, which are vincristine-resistant clones, are transplanted intraperitoneally into mice. When used together, the life is extended by 112 to 131%. Verapamil, an existing calcium antagonist, is 121-130% in combination with an antitumor agent
Life prolonged. Calcium antagonists such as verapamil and nicardipine have been reported to have a resistance overcoming effect, but their calcium antagonism is very strong. On the other hand, the compound of the present invention is characterized in that the calcium antagonism is much weaker than that of the above calcium antagonist. For example, the sixth
As is clear from the table, the compound of the present invention has a very weak action as compared to verapamil or nicardipine, and even at 10 ^-7 mol, almost no contraction-inhibiting action was observed. Further, verapamil requires 30 to 60 mg / kg (intraperitoneal administration) as shown in Table 5 to obtain the effect of overcoming tolerance, but its blood pressure lowering effect as a calcium antagonist is 0.2 to 0.4 mg / kg.
It is expressed at mg / kg (intravenous administration), and when verapamil is used, a stronger blood pressure lowering effect than the resistance overcoming effect appears as a side effect. Verapamil at 75 mg / kg has killed as many as 22 of 24 animals. In view of the above, combined administration of a compound of the present invention and an antitumor agent has fewer side effects and more effective effect of overcoming resistant cancer than administration of a calcium antagonist such as verapamil or nicardipine in combination with an antitumor agent. can get.

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Claims (5)

(57) [Claims] (1) Formula (I) (In the formula, R represents a pyridyl group, an N-morpholyl group, or an N-benzyl-N-methylamino group which may be substituted with one methyl group or ethyl group, and n represents an integer of 1 to 4. 1,)-dihydropyridine derivative represented by the formula: 2. The 1,4-dihydropyridine derivative according to claim 1, wherein R is a pyridyl group optionally substituted by one methyl group or ethyl group, and n is an integer of 1 to 3. . 3. The method according to claim 2, wherein R is a 2-pyridyl group, a 3-pyridyl group or a 6-methyl-2-pyridyl group.
The 1,4-dihydropyridine derivative according to the above item. 4. The 1,4-dihydropyridine derivative according to claim 1, wherein R is an N-morpholyl group and n is an integer of 1 to 3. 5. The 1,4-dihydropyridine derivative according to claim 1, wherein R is an N-benzyl-N-methylamino group and n is an integer of 1 to 3.