WO1997028152A1 - Dihydropyridine compounds and medicinal composition comprising the same - Google Patents

Abstract

Dihydropyridine compounds represented by general formula (I) or pharmacologically acceptable salts thereof and an anticancer tolerance restorative or an anticancer agent potentiator comprising the same as the active ingredient; wherein R1 represents -A-(3-pyridyl) (wherein A represents linear C¿3-6? alkylene); R?2¿ represents C¿2-6? alkyl, alkenyl or alkynyl, optionally substituted lower alkyl or lower alkenyl, or optionally substituted cycloalkyl; R?3¿ represents a group having one or two optionally substituted phenyl groups; and n is an integer of from 1 to 4.

Description

 TECHNICAL FIELD The present invention relates to dihydroxypyridine compounds and pharmaceutical compositions containing the same.

 The present invention relates to a novel dihydropyridin compound having an effect of overcoming an anticancer drug resistance or an effect of enhancing an anticancer agent effect, and a pharmaceutical composition comprising the compound or a pharmacologically acceptable salt or hydrate thereof and a carrier. In particular, the present invention relates to an anticancer drug resistance overcoming agent or an anticancer agent effect enhancer. Background art

 Many compounds are already known as dihydroxypyridine derivatives. Most of these known dihydropyridine derivatives have pharmacological activity on the circulatory system, and few other pharmacological activities have anti-inflammatory activity, hepatoprotective activity, etc. It is just that.

 On the other hand, at present, in cancer chemotherapy, "acquisition resistance", in which the effects of anticancer drugs are lost during treatment, has become a problem, and multidrug resistance, which is resistant to various anticancer drugs, has become an important problem . As a method of overcoming this multidrug resistance, it has been reported that combined administration of anticancer drugs and some calcium antagonists (such as dihydropyridin compounds such as dicardipine) has been reported to be effective. ., 41, 1967-1972 (1981), Cancer and Chemotherapy, Vol. 11, 750-759 (1984)].

Further, JP-A-2-40383, JP-A-2-240081, JP-B-6-92391, and JP-B-6-92401 disclose a dioxen ring or a dichain at the 4-position of dihydropyridine. Compounds having a ring bonded thereto are disclosed in JP-A-5-117235 and JP-A-2-138221. It is described that a compound in which an aromatic ring such as a phenyl group is bonded to the 4-position of dropyridine has an action of overcoming anticancer drug resistance.

 However, the invention described in the above Cancer Res., 41, 1967-1972 (1981) and Cancer and Chemotherapy, Vol. 11, 750-759 (1984) disclose that a drug having a calcium pathway-blocking action is an anticancer drug resistance overcoming drug. The disadvantage is that it is not always practical in terms of side effects. In other words, since calcium pathway blockers are potent drugs by nature, they act on the heart and blood vessels, etc., even in very small amounts. The disadvantage is that unavoidable effects are unavoidable.

 As described in JP-A-2-40383 and JP-A-2-240081, dihydropyridines described in the above-mentioned publications have an anticancer agent effect enhancing effect or an anticancer agent resistance overcoming effect. In addition, some compounds have some favorable properties because they have almost no calcium pathway-blocking effect, and these compounds are not sufficiently satisfactory in terms of the effect of overcoming the anticancer drug resistance. Furthermore, the dihydropyridin compounds described in these publications each have, as a substituent at the 4-position, a heterocyclic group such as a dioxene ring or a dichen ring or an aromatic ring group such as a phenyl group. There is no description of compounds having an alkyl group, an alkenyl group or a cycloalkyl group. Disclosure of the invention

Accordingly, an object of the present invention is to provide a compound having an effect of significantly increasing the sensitivity of cancer cells to an anticancer drug, particularly the sensitivity of cancer cells that have acquired resistance to the anticancer drug (an effect of overcoming anticancer drug resistance), or an effect of enhancing the effect of the anticancer agent. An object of the present invention is to provide a compound having (an anticancer drug effect enhancing action). Another object of the present invention is to provide a compound which exhibits an effect of extending the survival time of a tumor-bearing animal when used in combination with an anticancer agent, has little or no blocking action on the calcium pathway, and has low toxicity.

 According to the present invention, formula (I):

(Wherein, R ¹ represents one A— (3-pyridyl), A represents a C ₃ -C e linear alkylene group, R ² represents a C ₂ -C 6 alkyl group, an alkenyl group Or an alkynyl group, a lower alkyl group or a lower alkenyl group having a substituent, or a cycloalkyl group optionally having a substituent, and R ³ represents a phenyl group optionally having a substituent. Or a pharmacologically acceptable salt or hydrate thereof as an active ingredient, wherein n represents an integer of 1 to 4. And, in particular, an anticancer drug resistance overcoming agent or an anticancer agent effect enhancer. BEST MODE FOR CARRYING OUT THE INVENTION

 Hereinafter, the present invention will be described in more detail.

Preferred examples of R ¹ include groups in which A is a trimethylene group, a tetramethylene group, and particularly a group in which A is a trimethylene group.

R ³ represents a group having one or two fluorinated groups which may have a substituent such as a methoxy group, a methyl group, an ethyl group, a halogen atom and a nitro group. Preferred examples of R ³ include a group having one or two phenyl groups, specifically, a benzhydryl group, a benzyl group, and a phenyl group. And a diphenylacetyl group (benzhydrylcarbonyl group), in particular, a benzhydryl group.

n represents an integer of 1 to 4, preferably an integer of 2 or 3. Preferred examples of R ² include a C ₃ -C ₆ alkyl group, an alkenyl group, an alkynyl group, a lower (eg, C, mono) alkyl group or a cycloalkyl group substituted by a phenyl group. Can be. Specifically, n-propyl, iso-propyl, n-butyl, n-pentyl, n-hexyl, 1-methylpropyl, 2-methylpropyl, 1-methylbutyl Groups, 1-ethylpropyl group, 2,2-dimethylpropyl group, 2-methyl-1-propenyl group, cyclopropyl group, cyclopentyl group, cyclohexyl group, etc. Can be done.

 In the present invention, specific examples of preferable compounds include Compounds 1 to 12 described in the following Examples.

 The 1,4-dihydropyridine compound represented by the formula (I) provided by the present invention can be any of the well-known 1,4-dihydropyridine compounds conventionally used in the production of 1,4-dihydropyridine compounds. It can be manufactured according to the method.

That is, the compound represented by the formula (I) is composed of an aldehyde represented by the formula (と), an acetate acetate represented by the formula (お よ び) and a 3-amino compound represented by the formula (IV) A method in which a crotonic acid ester is reacted in the presence or absence of an organic solvent (method A), or an aldehyde represented by the formula (II) and an acetate represented by the formula (V) It can be produced by, for example, a method (method B) in which an ester and a 3-amino carboxylic acid ester represented by the formula (VI) are reacted in the presence or absence of an organic solvent.

 Compound (I)

 (Method B)

) = CHCOOR

 > Compound (I)

(Wherein, R ¹ , R ² , R ^3, etc. have the same meanings as those of the formula (I). The reaction used in these production methods is a known reaction conventionally used in the production of dihydropyridin compounds. (For example, the methods described in JP-B-46-40625, JP-B-56-37225, JP-A-60-214786, etc.). In addition to the above-mentioned method, the dropyridine compound can also be produced by appropriately applying other reactions described in these known documents. Are known compounds, which can be easily obtained or manufactured by those skilled in the art as required, for example, acetate acetate is obtained by reacting diketene with alcohols. It can be manufactured by , 3-Aminocrotonates can be produced by reacting the above acetate acetate with ammonia gas.Aldehydes can be produced by a known method widely used in their synthesis. The compound of formula (I) obtained by the present method, which can be easily produced by reduction of certain esters or oxidation of alcohols, can be obtained by a known treatment method (eg, extraction, chromatography, recrystallization, etc.). Separation, purification CT / JP96 / 02590.

The dihydropyridine compound represented by the formula (I) has an asymmetric carbon atom and thus has an optical isomer. In the present invention, any of the optical isomers and a mixture of the isomers are included. Further, the compound in which R ² is an alkenyl group may have a geometric isomer in addition to the optical isomer in some cases. In the present invention, both isomers and a mixture of isomers are included. In addition, the isomer mixture can be separated into the respective isomers by a fractional crystallization method or chromatography, if necessary.

 The compound according to the present invention exhibits an effect of enhancing the effect of an anticancer agent, further exhibits an effect of overcoming the anticancer agent resistance against adriamicin-resistant cancer and vincristine-resistant cancer, and shows the effect of using a cancer-bearing animal in combination with an anticancer agent. It is useful as an anticancer drug resistance overcoming agent or an anticancer drug effect enhancer because it extends the survival time.

 When the compound according to the present invention is used as an anticancer drug resistance overcoming agent or an anticancer agent effect enhancer, it can be administered by an appropriate administration method such as oral or parenteral. Oral dosage forms include, for example, tablets, granules, capsules, pills, powders, and liquids. Parenteral dosage forms include, for example, injections, suppositories, and the like. Can be These preparations can be prepared by a conventional method using the compound of the present invention or a pharmacologically acceptable salt thereof and a usual preparation carrier.

For example, in the case of oral preparations, excipients such as lactose, glucose, corn starch, and sucrose, disintegrants such as calcium carboxymethylcellulose and quinopenyl cellulose, and calcium stearate Lubricants such as magnesium stearate, tanolek, polyethylene glycol, hydrogenated oil, hydroxypropylcellulose, hydroxypropylmethylcellulose, carboxymethylcellulose, polypropylene Nyl alcohol, gelatin, arabic gum, and other binders, glycerin, wetting agents such as ethylene glycol, and other surfactants and flavoring agents as needed to achieve the desired dosage form. Can be prepared.

 In the case of parenteral preparations, diluents such as water, ethanol, glycerin, propylene glycol, polyethylene glycol, agar, and tragarant gum are used. If necessary, a dissolution aid, a buffer, a preservative, a fragrance, a coloring agent, and the like can be used.

 When the compound of the present invention is formulated as an anticancer drug resistance overcoming agent or an anticancer drug effect enhancer, the dosage unit of the compound of the present invention is 5 to 1,000 mg per adult per day for oral administration and 5 to 1,000 mg per day for oral administration. 5 to 200 mg, preferably 1 to 500 mg / day, preferably 1 to 200 mg / day for parenteral administration, and a desired therapeutic effect can be expected by each 1 to 3 times a day in divided doses. . Example

 Next, Synthesis Examples, Formulation Examples, and Test Examples of the compound according to the present invention are shown as Examples.

 (Synthesis example)

 A synthesis example is shown below, and the NMR data shows a signal of NMR-NMR measured in a CDCh solvent.

Example 1

 2, 6 — dimethyl 1 4 — η — pentyl 1, 4-dihydroxypyridin-3, 5 — dicarboxylic acid 3 — [3 — (4 — benzylpiperazine 1 11 yl) propyl] Synthesis of Ester, 5— [3— (3—Pyridyl) propyl] Ester (Compound 1)

Acetate acetic acid 3 — (4 — benzylpiperazine-1 1 — yl) propyl 3.2 g of the ester and 2.2 g of aminocrotonic acid 3-(3-pyridyl) propyl ester and caprolaldehyde l. Lg are heated under reflux in 10 ml of isopropanol for 7 hours. After completion of the reaction, the reaction solution is concentrated to dryness under reduced pressure, the residue is dissolved in 1N hydrochloric acid, washed with ethyl acetate, made alkaline with sodium hydroxide, and extracted with ethyl acetate. . The extracted solution is washed with water, dried over anhydrous sodium sulfate, and dried under reduced pressure. The oily substance was purified by silica gel chromatography to obtain 3.9 g (65.0%) of the target compound.

 NMR: 0.83 (3H, t), 1.20-1.31 (8H, m), 1.85 (2H, m), 2.00 (2H, m), 2.28 (3H, s), 2.29 (3H, s), 2.42-2.46 ( 2.73 (2H, t), 3.49 (2H, s), 3.94 (lH, t), 4.10-4.20 (4H, m), 5.68 (1H, br), 7.18-7.3 l (6H, m) ), 7.51 (lH, m), 8.43-8.46 (2H, m)

Example 2

 2, 6 — Dimethyl 1 4 1 n — Pentyl-1, 4 — Dihydroxypyridin 1, 3, 5 — Dicarboxylic acid 3 — [3 — (4 Benzhydrinolepiperazine 1 1 — yl) Synthesis of [Propyl] ester, 5— [3— (3-pyridyl) propyl] ester (Compound 2)

 Acetate acetic acid 3-(3-pyridyl) propyl ester 2.2 g, amino crotonic acid 3-(4-benzhydrinolepiperazine-1 1 -yl) 4.0 g and caprolaldehyde l.lg were reacted in 20 ml of ethanol at room temperature for 3 days. After completion of the reaction, the reaction solution is concentrated to dryness under reduced pressure, and extracted with ethyl acetate. The extracted solution is washed with water, dried over anhydrous sodium sulfate, and dried under reduced pressure. The oily substance was purified by silica gel chromatography to obtain 5.5 g (80.9%) of the target compound.

NR: 0.82 (3H, 0, 1.25 (8H, m), 1.84 (2H, m), 1.99 (2H, m), 2.27 (3H, s), 2.29 (3H, s), 2.44 (10H, m), 2.72 (2H, t) ヽ 3.94 (lH, t), 4. 13 (4H, m), 4.20 (lH, s), 5.71 (1H, br), 7.15-7.27 (7H, m), 7.40 (4H, d), 7.49 (lH, d), 8.42 (1H, d ), 8.45 (1H, s)

 The compounds of each of the examples synthesized according to Example 1 or Example 2 are listed below together with the starting materials used and the analysis values of NMR. The purification of the compound was carried out by recrystallizing the obtained crude substance with an appropriate solvent or, if necessary, by subjecting it to silica gel chromatography chromatography. Three

 2, 6 — dimethyl 4- 1-isopropyl-1, 4, 4-dihydroxypyridine 3, 5 — dicarboxylic acid 3 — [3-(4 — benzhydrinolepiperazine 1 1 yl) ) Synthesis of [propyl] ester, 5— [3— (3—pyridyl) propyl] ester (Compound 3)

 material :

 Acetate acetic acid 3 — (4 — Benzhydrinolepiperazine)

 Aminocrotonic acid 3 — (3 — pyridyl) propyl ester isobutyl aldehyde

 NMR: 0.74 (3H, d), 0.76 (3H, d), 1.60 (1H, m), 1.84 (2H, m), 1.99 (2H, m), 2, 30 (3H, s), 2.31 (3H, s), 2.39 (10H, m), 2.73 (2H, t), 3.93 (lH, d), 4.13 (4H, m), 4.19 (lH, s), 5.60 (1H, br), 7.15-7.28 (7H, m), 7.40 (4H, d), 7.49 (lH, d), 8.42 (lH, m), 8.46 (1H, d)

Example 4

 2, 6 — dimethyl 4 — n — butyl 1, 4 — dihydropyridin — 3, 5 — dicarboxylic acid 3 — [3-(4 — benzhydrinolepiperazine 1 1 yl) Synthesis of [Propyl] ester, 5— [3— (3—Pyridyl) propyl] ester (Compound 4)

material : Acetate acetic acid 3 — (4-benzoyl benzene)

 Aminocrotonic acid 3 — (3 — pyridyl) propyl ester n — Barrel aldehyde

 NMR: 0.83 (3H, t), 1.20 (4H, m), 1.33 (2H, m), 1.84 (2H, m), 1.99 (2H, m), 2.28 (3H, s), 2.29 (3H, s) , 2.39 (10H, m), 2.72 (2H, t), 3.94 (1H, t), 4.15 (4H, m), 4.20 (lH, s), 5.67 (1H, br), 7.15-7.28 (7H , m), 7.40 (4H, d), 7.49 (lH, d), 8.42 (1H, m), 8.46 (lH, d)

Example 5

 2, 6 — Dimethylyl 4 _ n — Pentyl-1, 4 — Dihydropyridin-1,3, 5 — Dicarboxylic acid 3 — [3-(4 — Difenylacetylpiperazine-1 1 —yl) Synthesis of [Propyl] ester, 5— [3- (3-pyridyl) propyl] ester (Compound 5)

 material :

 Acetate acetic acid 3 — (4 — diphenylacetylpiperazine — 1 — yl) propyl ester

 Aminocrotonic acid 3 — (3 — pyridyl) propyl ester Caproaldehyde

 NMR: 0.82 (3H, t), 1.19-1.29 (8H, m), 1.80 (2H, m), 1.19 (2H, m), 2.13 (2H, m). 2.24 (3H, s), 2.25 (3H, s), 2.37 (4H, m), 2.72 (2H, t), 3, 42 (2H, m), 3.67 (2H, m), 3.93 (lH, t), 4.14 (4H, m), 5.20 (lH , S), 6.26 (1H, br), 7.20-7.32 (11H, m), 7.51 (lH, d), 8.45 (2H, m) Example 6

2, 6-dimethyl-4, n-pentyl-1,4-dihydroxypyridine-1,3, 5-dicarboxylic acid 3— [3— (4-phenylbiperazine-1-1, yl) Synthesis of [Propyl] ester, 5— [3— (3-pyridyl) propyl] ester (Compound 6) Hara:

 Acetate acetic acid 3 — (4-phenylbiperazine)

 Aminocrotonic acid 3 — (3 — pyridyl) propyl ester Caproaldehyde

NMR: 0.84 (3H, t), 1.27 (8H, m), 1.94 (2H, m), 2.03 (2H, m), 2.30 (6H, s) _s 2.50 (2H, t), 2.58 (4H, m) , 2.74 (2H, t), 3.17 (4H, m), 3.97 (1H, t), 4.18 (4H, m), 5.60 (1H, br), 6.88 (3H, m), 7.24 (3H, m), 7.52 (1H, d), 8.45 (lH, m), 8.47 (1H, d)

Example 7

 2,6—dimethyl 4- (3—methyl 2—butyl) 1—4—dihydroxypyridine 3,5—dicarboxylic acid 3— [3— (3—pyridyl) propyl] ester Synthesis of, 5— [3— (4—benzhydrinolepiperazine-1 1-yl) propyl] ester (Compound 7)

 material :

 Acetate acetic acid 3 — (3 — pyridyl) propyl ester

 Aminocrotonic acid 3 — (4 — Benzhydrinolepiperazine) 1-propyl ester

 I Sovarel Aldehid

 Picture: 0.87 (6H, q), 1.13 (2H, t), 1.55 (lH, m), 1.84 (2H, m), 1.97

(2H, m), 2.28 (3H, s), 2.29 (3H, s), 2.43 (10H, m), 2.72 (2H, m), 3.

96 (1H, t), 4.13 (4H, m), 4.19 (lH, s), 7.14 to 8.45 (14H, m)

Example 8

2, 6 — dimethyl 4 — (3 — methyl 2 — butenyl) 1, 4 — dihydroxypyridine 1 3, 5 — dicarboxylic acid 3 — [3-(3 — pyridyl) propyl ] Ester, 5— [3— (4—Benzhydrinolepiperazine-1 1-yl) propyl] Ester (Compound 8) Synthesis material :

 Acetate acetic acid 3 — (3 — pyridyl) propyl ester

 Aminocrotonic acid 3 — (4 — Benzhydrinolepiperazine 1 — yl) propyl ester

 3 — methyl 2 — butenal

 NMR: 1.60 (3H, s), 1.77 (3H, s), 1.83 (2H, m), 1.98 (2H, m), 2.27

(3H, s), 2.28 (3H, s), 2.41 (10H, m), 2.69 (2H, m), 4.14 (4H, m), 4.

19 (lH, s), 4.59 (lH, d), 4.95 (lH, d), 7.15 to 8.45 (14H, m)

Example 9

 2,6—Dimethyl-1-cyclopropyl-1,4, -Dihydropyridin-1,3,5-Dicarboxylic acid 3— [3— (3—Pyridyl) propyl] ester, 5 —Synthesis of [3— (4-benzhydrinolepiperazine-1 1-yl) propyl] ester (Compound 9)

 material :

 Acetate acetic acid 3 — (3 — pyridyl) propyl ester

 Aminocrotonic acid 3 — (4 — Benzhydrinolepiperazin-1-yl) propyl ester

 Cyclopro pilaldehyde

 Hidden: 0.23 (4H, d), 0.82 (1H, m), 1.84 (2H, m), 1.99 (2H, m), 2.30

(3H, s), 2.31 (3H, s) _s 2.44 (10H, m), 2.72 (2H, m), 3.67 (lH, d), 4.

15 (4H, m), 4.19 (lH, s), 7.14 to 8.46 (14H, m)

Example 10

 2,6—Dimethylyl 4-cyclohexyl-1,4—Dihydroxypyridin-1,3,5-Dicarboxylic acid 3— [3- (3—Pyridyl) propyl] ester, 5— [3 —Synthesis of (4—benzhydrylpyrazin-1-yl) propyl] ester (Compound 10)

material : Acetoacetic acid 3 — (3 — pyridyl) propyl ester aminocrotonic acid 3 — (4 — benzhydryl pyridine 1 -yl) propyl ester

 Cyclohexylaldehyde

 Marauders: 0.81 to 1.64 (11H, m), 1.84 (2H, m), 1.99 (2H, m), 2.29 (3H, s), 2.31 (3H, s), 2.44 (10H, m), 2.73 (2H, m m), 3.93 (lH, d), 4.12 (4

H, m), 4.19 (lH, s), 7.14-8.46 (14H, m)

Example 11

 2,6—Dimethyl-1-4 (2—phenylethyl) 1-1,4—Dihydroxypyridin-1,3,5—Dicarboxylic acid 3— [3— (3—Pyridyl) propyl] ester Synthesis of, 5 — [3 — (4 — Benzhydrylpyrazine — 1-yl) propyl] ester (Compound 11)

 material :

 Acetate acetic acid 3 — (3 — pyridyl) propyl ester

 Aminocrotonic acid 3 — (4 Benzhydrinolepiperazin-1-yl) Propyl ester

 Neudro Sinnam Aldehid

 Hidden: 1.68 (2H, m), 1.84 (2H, m), 1.98 (2H, m), 2.29 (3H, s), 2.31

(3H, s), 2.44 (10H, m), 2.54 (2H, m), 2.71 (2H, m), 4.11 (1H, t), 4.

17 (4H, m), 4.20 (lH, s), 7.11-8.44 (19H, m)

Example 12

 2, 6 — dimethyl 1 4 — (n-pentyl) 1 1, 4 — dihydroxypyridin 1, 3, 5-dicarboxylic acid 3-[3-(3-pyridyl) propyl] ester Synthesis of, 5— [2— (4-benzylhydrazine) 1-yl) ethyl] ester (Compound 12)

 material :

Acetate acetic acid 3 — (3 — pyridyl) propyl ester Aminocrotonic acid 2 — (4 — Benzhydrinolepiperazine)

 Capron aldehyde

 NMR: 0.82 (3H, t), 1.18 to 1.30 (8H, m), 1.95 (2H, m), 2.28 (3H, s), 2.29 (3H, s), 2.44 to 2.54 (10H, m), 2, 69 (2H, m), 3, 93 (1H, t), 4.07 (lH, m), 4.12 (lH, m), 4.18 (1H, s), 4.25 (2H, m), 7, 15 to 8 · 46 (14H, m)

Example 13

 25 mg of compound 2 was dissolved in ethanol and adjusted to 1 ingZml. Inject 20〃1 of the sample solution into a CHIRACEL 0D column (Daicel Chemical Industry Co., Ltd., connecting two 4.0 mm Φ X 250 mm tubes) under the following conditions [mobile phase: hexane / isopropyl alcohol = 9 Z1, detection: 230 nm, flow rate: 0.5 ml / min] to separate optical isomers. Holding time about 100 ~

At 110 minutes (about 104 minutes peak) and about 110-120 minutes (about 115 minutes peak), each enantiomer, which is an optical isomer, eluted.

 (Formulation example)

Example 14 (Preparation of tablet)

 Compound of the present invention (Compound 3) 25 g

 Lactose 62 g

 Corn starch 40 g

 Hydrokin Pro pill cellulose 2 g

 Magnesium stearate 1

After the above-mentioned compound of the present invention, lactose and constarch are mixed until uniform, a 5 W / V% ethanol solution of hydroxyquine propylcellulose is added, and the mixture is kneaded and granulated. After passing through a 16-mesh sieve and sieving, the tablets were compressed in a conventional manner to give tablets each having a weight of 130 mg, a diameter of 7 mm, and a main drug content of 25 mg. (Test example)

Test example 1

 In vitro (in vitro) test of cancer cell growth inhibitory effect by combined use with doxorubicin

 Human nasopharyngeal carcinoma-derived KB cells (susceptible cells) and their multidrug-resistant clones, VJ-300 cells (resistant cells), were used as test cells. The culture solution was 10% fetal bovine serum (Flow Laboratories) and 0.29 mg Zml L-Glumin (Fab Laboratories) containing Eagle MEM medium (Yuisui Pharmaceutical Co., Ltd.). Manufactured). The test of the combined use of the test compound and doxorubicin (adriamycin, ADM), which is an anticancer drug, for the effect of overcoming the anticancer drug resistance or enhancing the effect of the anticancer drug was carried out as follows.

Suspend each test cell in the culture medium and adjust the cell density to about 200 cells / ml. The cell suspension was dispensed in a 2 ml Zutsushi petri dish, and cultured for 24 hours at 37 ° C in carbonate gas wells down Kyubeta one (5% C0 _2, 95% air). Thereafter, add 5 to 10 1 of each of the ADM aqueous solution of the specified concentration and the DMS0 solution of the test compound of the specified concentration, and continue culturing for another 7 days. After completion of the culture, the cells were fixed with methanol, stained with Giemsa, the number of colonies in each dish was counted, and a volume response curve was prepared. From this calculate the 50% ADM concentration of cell viability (LD _{5 D),} to determine the anticancer drug resistance overcoming effect及beauty anticancer effect enhancing effect. LD ₅ of ADM results in ADM alone group in KB cells. The concentration is shown as resistance level 1 and each LD _{5 is} shown below. The degree of resistance was calculated using the concentration as a relative ratio and is shown in Tables 1 and 2. In the table, ADM alone (control) refers to the ADM alone group, ADM + Compound 1 refers to the ADM + Compound 1 (1 / zg / ml) combination group, and similarly ADM + Compound 12 refers to ADM and Compound 12 (1 U g / mL). ) Represents the combination group. The results obtained using verapamil and dicardipine as reference compounds are shown in the table. In the table, ADM + verapamil refers to the combination group of ADM and verapaminole (1 fig / ml), and ADM + dicardipine. Represents the combination of ADM and dicardipine (3M)

Test example 2

In vitro study of the inhibitory effect of cancer cells on growth in combination with vincristine (i (n vitro) test

 In the same manner as in Test Example 1, a test was performed using bincristin (VCR) as an anticancer drug, a dose response curve was prepared, and the degree of resistance was calculated. The results are shown in Table 3. In the table, VCR alone (control) represents the VCR alone group, and VCR + compound 2 represents the VCR + compound 2 (I fig / ml) combined group. In addition, the results of using veravamil as a control compound are shown in the table. In the table, VCR + veravamil represents the combination group of VCR and verapa minole (1 gZml).

試験例 3

Test example 3

 In vivo test of the inhibitory effect of cancer cells on proliferation by combined use with anticancer drugs

 Anti-drug resistance overcoming effect in vincristine (VCR) resistant mice with leukemia cells

Group Six CDF, 10 ⁶ VCR resistant murine leukemia (P3 88 / VCR) cells to male mice transplanted intraperitoneally, the compound of the present invention UOOmgZkg) and VCR (100 ^ g / kg) for 5 days i.p. After the administration, the number of days of survival was determined, and the survival rate (TZ C) relative to the control was determined. The anticancer drug resistance overcoming effect (TZV) was determined by the following equation.

 Anticancer resistance overcoming effect (T / V)%

 Combination of VCR and the compound of the present invention

 Case life extension (TZ C)

 X 100

Life extension when using VCR alone (TZ C) VCR was dissolved in physiological saline, and the compound of the present invention was used by suspending it in a 0.5% carboxymethylcellulose sodium solution containing 0.1% Tween-80. The results are shown in Tables 4 and 5. In the table, control is the non-administration group, VCR alone is the VCR (100 g / kg) administration group, VCR + Compound 1 is the VCR (100 / zg / kg) and Compound 1 (100 mg / kg) combination group, and Similarly, VCR + Compound 6 represents the combination group of VCR (100 ^ g / kg) and Compound 6 (100 mg / kg).

 In addition, VCR + Compound 2 * marked with * indicates a group in which VCR (100 g Zkg) and Compound 2 (10 mg / kg) were used in combination.

 ¾ 4

 Table 5

 Test example 4

Action on chloride contraction in rat aorta Rats create a-ring specimens having a length of about 2 hidden from the thoracic aorta of Bok, 37 ° C, 95% 02 - a 5% C0 ₂ was suspended Magunu scan tube filled with aerated click Repusu solution. After stabilizing the sample for about 60 minutes, inject a potassium chloride solution into the Manugs tube to a final concentration of 50 M, and when the contraction reaction that occurs reaches equilibrium, a positive control compound or the compound of the present invention. was dissolved in dimethyl Chirusuruhokishi de were cumulatively injected at 1 X 10- '° M by Ri 1 X 10- ⁴ M or. The contraction force was recorded via FD-Pickup (Nihon Kohden) and polygraph (Nihon Kohden). The results are shown at 50% inhibitory concentration of high mosquito re um contraction in Table 6 (IC _5. Value).

 Table 6

試験例 5

Test example 5

 Acute toxicity test

 Animals used: ddY male mice (Japan SLC-1) 4-5 weeks old, 3-5 mice per group were used.

Test method: 0.5% carboxymethyl cellulose sodium (CMC-Na) containing 0.1% Tween 80 or 0.1% Tween 80 0.5% CMC_Na containing 10% ethanol containing the compound of the present invention And no deaths in 3-5 animals per group from 100 mgZkg to 1 / root 2 Mice until intraperitoneal injection. The survival of the animals is monitored up to 7 days after administration, and LDs are obtained by the Probit method. The value was calculated.

As a result, LD _{5 of} compound 2 was obtained. The value was over lOOOOmgZkg. Industrial applicability

 The dihydropyridine compound according to the present invention enhances its action when used in combination with an anticancer agent. The effect is particularly remarkable for clones that have acquired resistance to anticancer drugs. For example, as is clear from Table 3, VJ-300 cells, which are multidrug-resistant clones of KB cells derived from human nasopharyngeal carcinoma, are 709.1 times as large as cells that have not acquired resistance. The same effect (50% cell viability) cannot be obtained without using an anticancer agent at a concentration of 2%, whereas the same effect can be obtained at a concentration of 2.0 times with the compound 2 of the present invention (compound 1 g Zml). can get.

 In addition, the compounds of the present invention have low toxicity and weak calcium antagonism, and are effective in both in vitro (in vitro) and in vivo (in vivo) tests. Useful as an effect enhancer.

Claims

1. Formula (I)

Contract (Wherein, R ′ represents one A— (3-pyridyl), A represents a C ₃ -C 6 linear alkylene group, R ² represents a C ₂ -C e alkyl group, and A alkenyl group or an alkynyl group, a lower alkyl group or a lower alkenyl group having a substituent, or a cycloalkyl group which may have a substituent R ³ represents a group having one or two fluoro groups which may have a substituent, and n represents an integer of 1 to 4. Compound. 2. The dihydropyridine compound according to claim ¹ , wherein A of R ¹ is a trimethylene group. 3. The dihydropyridine compound according to claim 1 or 2, wherein R ³ represents a group having one or two phenyl groups, and n represents an integer of 2 or 3. 4. The dihydropyridinine compound according to claim ^3, wherein R ^{3 is a} benzhydryl group, a benzyl group, a phenyl group, or a diphenylacetyl group. 5. The dihydropyridine compound according to claim 4, wherein R ^{3 is a} benzhydryl group. 6. Any one of claims 1 to 5, wherein R ² is a C ₃ -C 6 alkyl group, alkenyl group or alkynyl group, a lower alkyl group substituted by a phenyl group, or a cycloalkyl group. Jihide described in item 1 Mouth pyridine compounds. 7. R ² n-propyl, isopropyl, n-butyl, n-pentyl, n-hexyl, 1-methylpropyl, 2-methylpropyl, 1_ Methylbutyl group, 1-ethylpropyl group, 2, 7. The dihydropyridine compound according to claim 6, which is a 2-dimethylpropyl group, a 2-methyl-11-propenyl group, a cyclopropyl group or a cyclohexyl group.  8. The dihydropyridine compound according to any one of claims 1 to 7 or a pharmacologically acceptable salt or hydrate thereof and a carrier. A pharmaceutical composition comprising:  9. A dihydropyridin compound or a pharmacologically acceptable salt or hydrate thereof according to any one of claims 1 to 7 and a carrier. An anticancer drug resistance overcoming agent.  10. A dihydropyridine compound or a pharmacologically acceptable salt or hydrate thereof according to any one of claims 1 to 7 and a carrier. Anticancer drug effect enhancer.