JPH05178858A - Dc-89 derivative - Google Patents

Abstract

PURPOSE:To provide the subject new compound useful as an antitumor agent. CONSTITUTION:A compound of formula {the group of formula II is group of formula III, etc.; W is H. group of formula IV [W<1>, W<2> are H, OR<4> (R<4> is lower alkyl, lower alkenyl)]}. The compound of formula I is obtained by treating a compound of formula V, etc., with a base (e.g. sodium methoxide, triethylamine or potassium carbonate) in an inert solvent.

Description

Detailed Description of the Invention

[0001]

This invention relates to DC-89 derivatives. The present compound exhibits excellent antitumor activity and is useful as an antitumor agent.

[0002]

2. Description of the Related Art Compounds similar to the DC-89 derivative of the present invention include DC-89A1 represented by the following structural formula:
DC-89A2, DC-89B1 and DC-89B2
Are known to exhibit antibacterial activity against various bacteria, as well as antitumor activity against melanoma B-16 and the like.

[0003]

[Chemical 8]

DC-89A1 is WO87 / 06265
, DC-89A2, DC-89B1 and DC-89
B2 is disclosed in Japanese Patent Laid-Open No. 2-119787. In addition, SF2582A and SF2582B, which are the same compounds as DC-89A2 and DC-89A1, are disclosed in JP-A No.
-139590. In connection with the compounds of the present invention, DC-88A having the structural formula:
No. 06265, DC113 is disclosed in JP-A-2-177890, which shows antibacterial activity against various bacteria and antitumor activity against melanoma B-16 and the like.

[0005]

[Chemical 9]

Further, DC-88A and DC-89 derivatives are disclosed in JP-A-2-288879, JP-A-3-7287 and JP-A-3-128379.
-1377741, Japanese Patent Application No. 3-186637 and Japanese Patent Application No. 4-20059. A derivative of SF2582C having a structure similar to that of the compound of the present invention is disclosed in JP-A-1-
275581 and CC-1065 and its derivatives are disclosed in JP-A-54-64695 and JP-A-60-19398.
9, WO88 / 04659 and EP-A-35945.
4, respectively.

[0007]

SUMMARY OF THE INVENTION An object of the present invention is to provide a DC-89 derivative having excellent antitumor activity.

[0008]

The present invention provides a compound of formula (I)

[0009]

[Chemical 10]

{Wherein

[0011]

[Chemical 11]

[0012]

[0013]

[Chemical 12]

Or

[0015]

[Chemical 13]

X represents a hydrogen atom or CO ₂ CH ₃
, Z is Cl or Br, R is a hydrogen atom, C
ONR ¹ R ² (wherein R ¹ and R ² are the same or different and represent a hydrogen atom, a lower alkyl group or a phenyl group) or

[0017]

[Chemical 14]

(In the formula, n represents an integer of 0 to 4, R ³ is CH ₂ ,

[0019]

[Chemical 15], Oxygen atom, N-CH₃Or N-CH₂ CONR¹ R
 ²Represents R¹ And R ² Is as defined above. ) Table
And W is a hydrogen atom or

[0020]

[Chemical 16]

[In the formula, W ¹ and W ² are the same or different and each represent a hydrogen atom or OR ⁴ (in the formula, R ⁴ represents a lower alkyl group or a lower alkenyl group). ]] Is represented. } The DC-89 derivative represented by these, or its pharmacologically acceptable salt. Hereinafter, the compound represented by formula (I) is referred to as compound (I). Similarly, the compounds represented by formulas (II) to (IV) are referred to as compounds (II) to (IV). The compounds (I) a and (I) b are meant to be included in the compound (I). R ¹ , R ² and R ^{4 of the} above formula (I)
In the definition of, a lower alkyl group means a linear or branched alkyl group having 1 to 4 carbon atoms, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, se
C-butyl, tert-butyl and the like are included, and in the definition of R ⁴ , the lower alkenyl group includes C 2-4 vinyl, allyl, propenyl, butenyl and the like. Next, the method for producing the compound (I) will be described. Method 1 Of the compound (I),

[0022]

[Chemical 17]

[0023]

[0024]

[Chemical 18]

And a compound (I) in which W is a hydrogen atom
a is the formula described in JP-A-3-128379.

[0026]

[Chemical 19]

Compound (A) represented by or a formula

[0028]

[Chemical 20]

The compound (B) represented by
It can be produced by treating with a base.

[0030]

[Chemical 21]

As the base, sodium methoxide, sodium hydroxide, potassium hydroxide, potassium t-butoxide, triethylamine, 1,8-diazabicycloundecene (DBU), potassium carbonate and the like are used. ) Or (B) is used in an amount of 1 to 3 equivalents.
As the inert solvent, water, methanol, tetrahydrofuran (THF), dioxane, acetonitrile, etc. may be used alone or in combination. The reaction is usually performed at -20 ° C to 50 ° C and is completed in 30 minutes to 5 hours.

Method 2 Of the compound (I),

[0033]

[Chemical formula 22]

Is

[0035]

[Chemical formula 23]

The compound (I) b in which W is a group other than a hydrogen atom is obtained by reacting the compound (I) a with a reactive derivative of the corresponding carboxylic acid in the presence of a base in an inert solvent. Can be manufactured by.

[0037]

[Chemical formula 24]

(In the formula, W ^a represents W other than ^a hydrogen atom,
X has the same meaning as above. ) As the base, sodium hydride, lithium diisopropylamide, potassium t-butoxy, triethylamine,
4-dimethylaminopyridine and the like are included, and usually used in 1 to 2 equivalents based on compound (I) a. As the inert solvent, dimethylformamide, THF, toluene, dimethylsulfoxide, etc. may be used alone or in combination. Reactive derivatives of carboxylic acids include acyl halides, acyl halides such as acid bromides, p-nitrophenyl esters, 2,4,5-trichlorophenyl esters, active esters such as N-oxysuccinimide esters, and the like. The reactive derivative is usually 1 to the compound (I) a.
3 equivalents are used. The reaction is carried out at -50 ° C to 30 ° C and is completed in 30 minutes to 1 day.

Method 3 Of the compound (I),

[0040]

[Chemical 25]

However,

[0042]

[Chemical formula 26]

Compound (I) wherein R is a hydrogen atom
c is a compound (I) a or (I) b in an inert solvent,
It can be produced by reacting hydrochloric acid or hydrobromic acid.

[0044]

[Chemical 27]

(In the formula, X, W and Z have the same meanings as described above.) Hydrochloric acid or hydrobromic acid is usually used in an amount of 1 to 20 equivalents based on compound (I) a or (I) b. As the inert solvent, water, dimethylformamide, THF, toluene, dioxane, acetonitrile, etc. may be used alone or in combination. The reaction is usually carried out at -20 ° C to 50 ° C and is completed in 10 minutes to 1 hour.

Method 4-1 Among compound (I),

[0047]

[Chemical 28]

Is

[0049]

[Chemical 29]

And R is CONR ¹ R ² or

[0051]

[Chemical 30]

Compound (I) d which is
In the presence of a base in an inert solvent, the formula

[0053]

[Chemical 31]

(Wherein R ¹ and R ² have the same meanings as described above) or

[0055]

[Chemical 32]

It can be produced by reacting the compound (II) represented by the formula (wherein R ³ and n have the same meanings as described above).

[0057]

[Chemical 33]

(Where R ^a is CONR ¹ R in the definition of R)
² or

[0059]

[Chemical 34]

And X, W and Z are as defined above. The base includes triethylamine, pyridine, 4-dimethylaminopyridine and the like and is usually used in an amount of 1 to 5 equivalents relative to compound (I) c, but it may also be used in a large excess also as a solvent. As the inert solvent, pyridine, methylene chloride, dimethylformamide, THF, toluene and the like can be used alone or in combination. Compound (II) is usually used in the amount of 1 to 10 equivalents relative to compound (I) c. The reaction is carried out at -50 ° C to 50 ° C and is completed in 30 minutes to 1 day.

Method 4-2 Compound (I) d can also be produced by the following steps. (Step 1) Compound (III) can be produced by reacting compound (I) c with p-nitrophenyl chloroformate in the presence of a base in an inert solvent.

[0062]

[Chemical 35]

(In the formula, X, Z and W have the same meanings as described above.) P-Nitrophenyl chloroformate is usually used in an amount of 1 to 5 equivalents based on compound (I) c. As an inert solvent, pyridine, methylene chloride, dimethylformamide, THF,
Toluene and the like are used alone or in combination. The reaction is carried out at -80 ° C to 50 ° C and is completed in 30 minutes to 1 day.

(Step 2) Compound (I) d is compound (II) d
I) in the presence of a base, in an inert solvent, the formula

[0065]

[Chemical 36]

(Wherein R ¹ and R ² are as defined above) or

[0067]

[Chemical 37]

It can be produced by reacting the compound (IV) represented by the formula (wherein R ³ and n have the same meanings as described above).

[0069]

[Chemical 38]

(In the formula, R ^a , X, Z and W have the same meanings as described above.) The base includes triethylamine, pyridine, 4-dimethylaminopyridine and the like, and is usually 1 to 1 with respect to compound (III). It is used in an amount of 5 equivalents, but may be used in a large excess also as a solvent. As the inert solvent, pyridine, methylene chloride, dimethylformamide, THF, toluene and the like can be used alone or in combination. Compound (IV) is usually used in an amount of 1 to 5 equivalents based on compound (III). The reaction is-
It is performed at 80 ° C to 50 ° C and is completed in 30 minutes to 1 day.

After completion of the reaction in each step, water, an acid, a buffer solution and the like are added to the reaction solution if necessary, and the mixture is extracted with a non-water-soluble solvent such as ethyl acetate, chloroform and ether. The extract is washed with water, brine, etc., dried over anhydrous sodium sulfate, etc., and the residue obtained after evaporation of the solvent is subjected to column chromatography on silica gel, thin layer chromatography, high performance liquid preparative chromatography, recrystallization. And so on.

When it is desired to obtain a salt of compound (I), when compound (I) can be obtained in the form of a salt, it can be purified as it is. When it is obtained in a free form, compound (I) can be obtained. May be dissolved or suspended in a suitable solvent, and a suitable acid may be added to form a salt. In addition, the intermediate can be used in the next step without isolation and purification after the reaction. The compound (I) or a pharmaceutically acceptable salt thereof may exist in the form of an adduct with water or various solvents, and these adducts are also included in the present invention. Further, all the possible isomers including the optically active form of the compound (I) and a mixture thereof are included in the present invention.

Table 1 shows the structures and compound numbers of representative compounds belonging to compound (I).

[0074]

[Table 1]

Next, the pharmacological activity of the representative compound (I) will be described with reference to experimental examples. Experimental Example 1. HeLaS ₃ cell growth inhibition test 3 in MEM medium containing 10% fetal calf serum and 2 mM glutamine in each well of a 96-well microtiter plate
HeLaS ₃ cells adjusted to × 10 ⁴ cells / ml were dispensed in 0.1 ml aliquots. 37 ° C overnight in carbon dioxide incubator
After culturing in, the compound (I) appropriately diluted with the medium was added to 0.05
ml was added to each well.

Incubate the cells in a carbon dioxide incubator.
After culturing at ℃ for 72 hours, remove the culture supernatant, wash once with phosphate buffered saline (PBS), add 0.1 ml of medium containing 0.02% neutral red to each well, and
The cells were stained by culturing in a carbon dioxide gas incubator at ℃ for 1 hour. After removing the culture supernatant, it was washed once with physiological saline, and the pigment was extracted with 0.001 N hydrochloric acid / 30% ethanol. The absorption of the extract at 550 nm was measured with a microplate reader, and the absorption of untreated cells and cells treated with a known concentration of compound (I) was compared to determine the drug concentration that inhibits cell growth by 50% (IC ₅₀ ) Was calculated.

[0077]2. Therapeutic effect on sarcoma 180 tumor
Fruit Salco to 5 male ddY mice weighing 18 to 20 g per group
Ma 180 tumor 5 × 10 ^Five Individuals were transplanted subcutaneously in the axilla. Transfer
1 day after planting, containing the compound (I) at the concentration shown in Table 2
0.2 ml of saline was administered intravenously. T / 7 days after transplantation
C [T: Average tumor volume of test example (mm³), C: control (raw
Mean tumor body of 0.2 ml of saline solution administered intravenously)
Product (mm³)] Was measured.

The respective results are shown in Table 2.

[0079]

[Table 2]

The acutely toxic compound (I) was intravenously administered to 20 ± 1 g of male ddY-type mice. MLD (minimum lethal dose) was determined by measuring the mortality rate 14 days after administration. The results are shown in Table 3.

[0081]

[Table 3]

The compound (I) or a pharmaceutically acceptable salt thereof can be used alone or together with at least one pharmaceutically acceptable auxiliary agent as an antitumor composition.
For example, compound (I) or a salt thereof is dissolved in physiological saline or an aqueous solution of glucose, lactose, mannitol or the like to give a pharmaceutical composition suitable as an injection. Alternatively, compound (I) or a salt thereof is freeze-dried according to a conventional method, and sodium chloride is added thereto to prepare a powder injection.

The present pharmaceutical composition may contain additives well known in the field of formulation, such as pharmaceutically acceptable salts, if necessary. The dose of the composition varies depending on the age, symptoms, etc. of the patient, but 0.01 to 30 mg / kg / day of Compound (I) is administered to mammals including humans. Administration is carried out, for example, once a day (single administration or daily administration) or intermittently by intravenous injection 1 to 3 times a week and once every 2 to 3 weeks. If desired, intraarterial administration, intraperitoneal administration, intrathoracic administration, etc. can be performed in the same dosage and administration form. If desired, oral administration is possible in the same dosage and administration form. Oral dosage forms include tablets, capsules, powders, granules, ampoules and the like, which contain the pharmaceutical auxiliaries well known in the pharmaceutical field.

Examples of the present invention will be shown below. The physicochemical properties shown in the following examples were measured by the following instruments. In addition, silica gel is Wako Gel C manufactured by Wako Pure Chemical Industries, Ltd.
-200 was used.

[0085]

【Example】

Example 1 Synthesis of Compound 1 To 14 mg (0.029 mmol) of compound (A), 2 ml of methanol and 28 wt% sodium methoxide (0.0.
0.03 ml of a methanol solution containing 16 mmol) was added, and the mixture was stirred at room temperature for 1 hour. The reaction mixture was adjusted to pH 7 with 0.
2M phosphate buffer was added, and the mixture was extracted with chloroform. The chloroform layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was subjected to silica gel chromatography (20 ml, chloroform:
Purified with methanol = 25: 1), 7.4 mg of compound 1
(Yield: 99%) was obtained.

The physicochemical properties of Compound 1 are as follows. ¹ H-NMR (400MHz, CDCl ₃ + DMSO-d ₆ ) δ (ppm); 11.72 (1H, br
s), 6.92 (1H, br s), 5.34 (1H, s), 3.72 (3H, s), 3.67 (1
H, dd, J = 10.6,5.3Hz), 3.51 (1H, dd, J = 10.6,10.6Hz), 3.39
(1H, m), 2.46 (3H, s), 1.98 (1H, dd, J = 7.7,2.4Hz), 0.98
(1H, dd, J = 4.6,2.9Hz) SIMS (m / z); 259 (M + H) ⁺ IR (KBr) υ (cm ^-1 ); 1682,1607,1573,1458,1379,1305,12
73,1229,1194,1156,1108

Example 2 Synthesis of compound 2 Example 2 was prepared by adding 2 ml of N, N-dimethylformamide to 23 mg (0.57 mmol) of 60% sodium hydride.
2 ml of N, N-dimethylformamide solution containing 95.6 mg (0.37 mmol) of the compound 1 obtained in
The mixture was stirred at 0 ° C. for 2 hours under an argon atmosphere. After the reaction solution was cooled to -50 ° C, 2 ml of N, N-dimethylformamide solution containing 166 mg (0.56 mmol) of 4-nitrophenyl ester of 4-methoxycinnamic acid was added,
The mixture was stirred at -50 ° C to room temperature for 1 hour. To this reaction mixture was added 0.2M phosphate buffer (pH 7), and the mixture was extracted with ethyl acetate. The ethyl acetate layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was purified by silica gel chromatography (20 ml, chloroform: methanol = 50: 1) to obtain 132 mg of compound 2 (yield: 85%).

The physicochemical properties of Compound 2 are as follows. ¹ H-NMR (400MHz, CDCl ₃ ) δ (ppm); 11.14 (1H, br), 7.79 (1H,
d, J = 15.4Hz), 7.52 (2H, d, J = 8.7Hz), 7.26 (1H, s), 6.91
(2H, d, J = 8.8Hz), 6.75 (1H, d, J = 15.4Hz), 4.24 (1H, dd, J =
10.9,10.9Hz), 4.15 (1H, dd, J = 10.9,4.8Hz), 3.85 (3H,
s), 3.82 (3H, s), 3.56 (1H, m), 2.62 (3H, s), 2.39 (1H, d
d, J = 7.6,3.5Hz), 1.31 (1H, dd, J = 4.9,3.5Hz) EIMS (m / z); 4
19 (M) ⁺ IR (KBr) υ (cm ^-1 ); 1702,1601,1512,1390,1292,1241,12
25,1173,1110,1072

Example 3 Synthesis of compound 3 30 mg (0.072 mm) of compound 2 obtained in example 2
3 ml of acetonitrile and 48% hydrobromic acid
1.5 ml was added, and the mixture was stirred at room temperature for 1 hour. 1N Hydrobromic acid was added to this reaction mixture, and the mixture was extracted with chloroform. The chloroform layer was dried over anhydrous sodium sulfate and then concentrated under reduced pressure. The crude product obtained was mixed with 3 ml of methylene chloride.
3 ml of a methylene chloride solution containing 36 mg (0.179 mmol) of p-nitrophenyl chloroformate was added at −78 ° C., and then triethylamine 0.02 was added.
5 ml (0.180 mmol) was added and the mixture was stirred for 0.5 hours.
0.032 ml of 50% dimethylamine aqueous solution was added to this reaction solution.
(0.356 mmol) was added, and the temperature was -78 ° C to room temperature.
Stir for 5 hours. To this reaction mixture was added 0.2M phosphate buffer (pH 7), and the mixture was extracted with chloroform. The chloroform layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was purified by silica gel chromatography (30 ml, chloroform: methanol = 100: 1) to obtain 36 mg of compound 3 (yield: 90%).

The physicochemical properties of Compound 3 are as follows. ¹ H-NMR (400MHz, CDCl ₃ ) δ (ppm); 8.98 (1H, br), 8.25 (1H,
s), 7.80 (1H, d, J = 15.5Hz), 7.56 (2H, d, J = 8.7Hz), 6.93
(2H, d, J = 8.8Hz), 6.81 (1H, d, J = 15.5Hz), 4.56 (1H, m),
4.48 (1H, br d, J = 10.6Hz), 4.30 (1H, dd, J = 9.1,2.4Hz),
3.94 (3H, s), 3.86 (3H, s), 3.81 (1H, dd, J = 10.4,1.7Hz),
3.34 (1H, dd, J = 10.5,10.4Hz), 3.18 (3H, s), 3.06 (3H, s),
2.60 (3H, s) SIMS (m / z); 572,570 (M + H) ⁺ IR (KBr) υ (cm ^-1 ); 2364,1701,1686,1637,1601,1511,13
99,1250,1173,1109,1091

Example 4 Synthesis of Compound 4 20 mg (0.048 mm) of Compound 2 obtained in Example 2
5 ml of acetonitrile and 1N hydrobromic acid.
2.2 ml was added, and the mixture was stirred at room temperature for 1 hour. 1N Hydrobromic acid was added to this reaction mixture, and the mixture was extracted with chloroform. The chloroform layer was dried over anhydrous sodium sulfate and then concentrated under reduced pressure. The crude product obtained was mixed with 3 ml of methylene chloride.
And p-nitrophenyl chloroformate 19.3 mg (0.096 mmol) was added at -10 ° C, and then triethylamine 0.013 ml (0.096 mmo).
1) was added and the mixture was stirred for 0.5 hours. Piperidine is added to this reaction solution.
0.014 ml (0.14 mmol) was added, and the mixture was stirred at -10 ° C to room temperature for 0.5 hr. The reaction mixture was adjusted to pH 7
0.2M phosphate buffer was added, and the mixture was extracted with chloroform.
The chloroform layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting crude product was purified by silica gel chromatography (30 ml, n-hexane: ethyl acetate = 1: 1) to give compound 4 (22 m).
g (yield: 75%) was obtained.

The physicochemical properties of Compound 4 are as follows. ¹ H-NMR (400MHz, CDCl ₃ ) δ (ppm); 9.21 (1H, br), 8.22 (1H,
s), 7.80 (1H, d, J = 15.2Hz), 7.57 (2H, d, J = 8.7Hz), 6.94
(2H, d, J = 8.8Hz), 6.81 (1H, d, J = 15.1Hz), 4.54 (1H, m),
4.47 (1H, br d, J = 10.5Hz), 4.31 (1H, dd, J = 9.2,9.2Hz),
3.95 (3H, s), 3.86 (3H, s), 3.80 (1H, dd, J = 10.2,2.2Hz),
3.68 (2H, br), 3.55 (2H, br), 3.21 (1H, dd, J = 10.2,10.2H
z), 2.53 (3H, s), 1.68 (6H, br) SIMS (m / z); 612,610 (M + H) ⁺ IR (KBr) υ (cm ^-1 ); 2930,2364,1694,1649,1599 , 1511,14
31,1408,1244,1215,1092,1025

Example 5 Synthesis of compound 5 20 mg (0.048 mm) of compound 2 obtained in example 2
5 ml of acetonitrile and 1N hydrobromic acid.
2.2 ml was added, and the mixture was stirred at room temperature for 1 hour. 1N Hydrobromic acid was added to this reaction mixture, and the mixture was extracted with chloroform. The chloroform layer was dried over anhydrous sodium sulfate and then concentrated under reduced pressure. The crude product obtained was mixed with 3 ml of methylene chloride.
And p-nitrophenyl chloroformate 19.3 mg (0.096 mmol) was added at -10 ° C, and then triethylamine 0.013 ml (0.096 mmo).
1) was added and the mixture was stirred for 0.5 hours. Pyrrolidine is added to this reaction solution.
0.012 ml (0.14 mmol) was added, and the mixture was stirred at -10 ° C to room temperature for 0.5 hr. The reaction mixture was adjusted to pH 7
0.2M phosphate buffer was added, and the mixture was extracted with chloroform.
The chloroform layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting crude product was purified by silica gel chromatography (30 ml, chloroform: methanol = 100: 1) to give compound 5 as 2
2 mg (yield: 76%) was obtained.

The physicochemical properties of Compound 5 are as follows. ¹ H-NMR (400MHz, CDCl ₃ ) δ (ppm); 8.92 (1H, br), 8.26 (1H,
s), 7.80 (1H, d, J = 15.2Hz), 7.57 (2H, d, J = 8.7Hz), 6.94
(2H, d, J = 8.8Hz), 6.80 (1H, d, J = 15.1Hz), 4.57 (1H, m),
4.47 (1H, br d, J = 10.3Hz), 4.30 (1H, dd, J = 9.0,9.0Hz),
3.96 (3H, s), 3.86 (3H, s), 3.81 (1H, dd, J = 10.2,2.1Hz),
3.65 (2H, t, J = 6.6Hz), 3.51 (2H, t, J = 6.6Hz), 3.22 (1H, d
d, J = 10.2,10.2Hz), 2.69 (3H, s), 2.00 (4H, br) SIMS (m / z); 598,596 (M + H) ⁺ IR (KBr) υ (cm ^-1 ); 2944, 1697,1637,1491,1412,1313,12
18,1109,1087

Example 6 Synthesis of compound 6 50 mg (0.12 mmo) of compound 2 obtained in example 2
l) in 5 ml of acetonitrile and 48% hydrobromic acid 2.
5 ml was added, and the mixture was stirred at room temperature for 1 hour. 1N Hydrobromic acid was added to this reaction mixture, and the mixture was extracted with chloroform.
The chloroform layer was dried over anhydrous sodium sulfate and then concentrated under reduced pressure. The obtained crude product was dissolved in 5 ml of methylene chloride, 5 ml of a methylene chloride solution containing 61 mg (0.30 mmol) of p-nitrophenyl chloroformate was added at -78 ° C, and then 0.042 ml of triethylamine was added.
(0.30 mmol) was added and the mixture was stirred for 0.5 hours. 0.040 ml (0.36 mmo) of N-methylpiperazine was added to this reaction solution.
1) was added, and the mixture was stirred at -78 ° C to room temperature for 0.5 hours.
To this reaction mixture was added 0.2M phosphate buffer pH7,
It was extracted with chloroform. The chloroform layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was purified by silica gel chromatography (30 ml, chloroform: methanol = 20: 1) to obtain 67 mg of compound 6 (yield: 89%).

The physicochemical properties of Compound 6 are as follows. ¹ H-NMR (400MHz, CDCl ₃ ) δ (ppm); 9.79 (1H, br), 8.21 (1H,
s), 7.77 (1H, d, J = 15.2Hz), 7.57 (2H, d, J = 8.8Hz), 6.93
(2H, d, J = 8.8Hz), 6.78 (1H, d, J = 15.3Hz), 4.53 (1H, m),
4.41 (1H, br d, J = 10.5Hz), 4.25 (1H, dd, J = 9.3,9.3Hz),
3.95 (2H, br), 3.93 (3H, s), 3.86 (3H, s), 3.84 (2H, br),
3.79 (1H, dd, J = 9.7,2.6Hz), 3.22 (1H, dd, J = 10.2,10.2H
z), 2.94 (4H, br), 2.67 (3H, s), 2.62 (3H, s) SIMS (m / z); 627,625 (M + H) ⁺ IR (KBr) υ (cm ^-1 ); 2360, 1692,1647,1602,1507,1400,12
91,1217,1173,1094

Example 7 Synthesis of compound 7 To 45 mg (0.072 mmol) of compound 6, 2 ml of ethanol and 4 ml of methanol were added, 0.025 ml of 5.8N hydrogen chloride-ethanol was added, and the mixture was stirred at 0 ° C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain 47 mg of compound 7.
(Yield: 99%) was obtained.

The physicochemical properties of compound 7 are as follows. ¹ H-NMR (400MHz, DMSO-d ₆ ) δ (ppm); 12.07 (1H, br), 10.57
(1H, br), 8.10 (1H, s), 7.74 (2H, d, J = 8.8Hz), 7.58 (1H,
d, J = 15.3Hz), 7.06 (1H, d, J = 15.3Hz), 7.00 (2H, d, J = 8.8H
z), 4.50 (1H, m), 4.42 (3H, br), 4.17 (1H, br), 3.85 (3H,
s), 3.82 (3H, s), 3.79 (1H, br), 3.58 (3H, br), 3.50 (4H,
br), 2.86 (3H, s), 2.68 (3H, s) IR (KBr) υ (cm ^-1 ); 2364,1740,1705,1648,1599,1511,14
05,1251,1218,1173,1095,1023

Example 8 Synthesis of Compound 8 25 mg (0.06 mmo) of Compound 2 obtained in Example 2
l) with 1.5 ml of acetonitrile and 48% hydrobromic acid
1.5 ml was added, and the mixture was stirred at room temperature for 1 hour. 1N Hydrobromic acid was added to this reaction mixture, and the mixture was extracted with chloroform. The chloroform layer was dried over anhydrous sodium sulfate and then concentrated under reduced pressure. The crude product obtained was treated with dichloromethane (2 m).
It was dissolved in 1 and added with 37 mg (0.18 mmol) of p-nitrophenyl chloroformate at −78 ° C., and then 0.025 ml (0.18 mmol) of triethylamine to give 0.5.
Stir for hours. Piperidinopiperidine 35 was added to this reaction solution.
mg (0.21 mmol) was added at -78 ° C to 0 ° C.
It was stirred for 0.5 hour. To this reaction mixture was added 0.2M phosphate buffer (pH 7), and the mixture was extracted with chloroform. The chloroform layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was purified by silica gel chromatography (20 ml, chloroform: methanol = 10: 1) to obtain 38 mg of compound 8 (yield:
91%) was obtained.

The physicochemical properties of compound 8 are as follows. ¹ H-NMR (270MHz, CDCl ₃ ) δ (ppm); 9.14 (1H, br), 8.23 (1H,
s), 7.79 (1H, d, J = 15.2Hz), 7.58 (2H, d, J = 8.6Hz), 6.94
(2H, d, J = 8.6Hz), 6.80 (1H, d, J = 15.2Hz), 4.57 (1H, m),
4.45 (2H, m), 4.33 (2H, m), 3.96 (3H, s), 3.86 (3H, s), 3.
79 (1H, br d, J = 9.7Hz), 3.22 (1H, dd, J = 10.0,10.0Hz), 3.
03 (1H, m), 2.88 (1H, m), 2.66 (3H, s), 2.62 (5H, br), 1.9
2 (2H, br), 1.67 (6H, br), 1.49 (2H, br) FABMS (m / z); 695,693 (M + H) ⁺ IR (KBr) υ (cm ^-1 ); 1701,1642,1599 , 1560,1542,1515,14
09,1253,1208,1092

Example 9 Synthesis of compound 9 To 26 mg (0.037 mmol) of compound 8, 1 ml of ethanol and 0.5 ml of methanol were added, and 6.86N hydrogen chloride-ethanol 0.008 ml was added, and the mixture was stirred at room temperature for 1 hour. .. The reaction mixture was concentrated under reduced pressure to give compound 9
mg (yield: 96%) was obtained.

The physicochemical properties of Compound 9 are as follows. ¹ H-NMR (270MHz, DMSO-d ₆ ) δ (ppm); 12.02 (1H, s), 9.96 (1
H, br), 8.05 (1H, s), 7.74 (2H, d, J = 8.9Hz), 7.57 (1H, d, J
= 15.3Hz), 7.05 (1H, d, J = 15.4Hz), 6.99 (1H, d, J = 8.9Hz),
4.46 (2H, m), 4.41 (2H, br), 4.19 (1H, br d, J = 13.3Hz),
3.84 (3H, s), 3.80 (3H, s), 3.77 (1H, br), 3.47 (5H, br),
3.13 (1H, br d, J = 12.9Hz), 2.95 (2H, br), 2.66 (3H, s),
2.15 (2H, br), 1.82 (6H, br), 1.70 (2H, br) IR (KBr) υ (cm ^-1 ); 1688,1646,1598,1514,1407,1252,12
13,1093,1023

Example 10 Synthesis of Compound 10 25 mg of Compound 2 obtained in Example 2 (0.06 mmo)
l) with 1.5 ml of acetonitrile and 48% hydrobromic acid
1.5 ml was added, and the mixture was stirred at room temperature for 1 hour. 1N Hydrobromic acid was added to this reaction mixture, and the mixture was extracted with chloroform. The chloroform layer was dried over anhydrous sodium sulfate and then concentrated under reduced pressure. The crude product obtained was treated with dichloromethane (2 m).
It was dissolved in 1 and added with 37 mg (0.18 mmol) of p-nitrophenyl chloroformate at -78 ° C and then 0.025 ml (0.18 mmol) of triethylamine, and stirred for 0.5 hours. To this reaction solution, 39 mg (0.21 mmol) of N-isopropyl-1-piperazine acetamide was added,
The mixture was stirred at −78 ° C. to 0 ° C. for 0.5 hours. To this reaction mixture was added 0.2M phosphate buffer (pH 7), and the mixture was extracted with chloroform. The chloroform layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was subjected to silica gel chromatography (20 ml,
Purification with chloroform: methanol = 20: 1) yielded 31 mg of compound 10 (yield: 73%).

The physicochemical properties of Compound 10 are as follows. ¹ H-NMR (270MHz, CDCl ₃ ) δ (ppm); 8.96 (1H, br s), 8.25 (1
H, s), 7.80 (1H, d, J = 15.5Hz), 7.57 (2H, d, J = 8.5Hz), 6.9
4 (2H, d, J = 8.5Hz), 6.80 (1H, d, J = 15.5Hz), 6.80 (1H, m),
4.58 (1H, m), 4.47 (1H, br d, J = 10.9Hz), 4.31 (1H, dd, J =
8.9,8.9Hz), 3.96 (3H, s), 3.86 (3H, s), 3.79 (2H, m), 3.
63 (2H, br), 3.23 (1H, dd, J = 10.0,10.0Hz), 3.05 (2H, s),
2.67 (3H, s), 2.61 (4H, br), 1.27 (1H, m), 1.19 (6H, d, J =
6.3Hz) FABMS (m / z); 712,710 (M + H) ⁺ IR (KBr) υ (cm ^-1 ); 1701,1646,1602,1561,1514,1409,13
05,1249,1219,1189,1093

Example 11 Synthesis of Compound 11 18 mg of Compound 10 obtained in Example 10 (0.025)
mmol) ethanol 1 ml and methanol 1 ml
Was added, and 6.86N hydrogen chloride-ethanol 0.006 ml was added.
Was added and stirred at room temperature for 3 hours. The reaction liquid was concentrated under reduced pressure to obtain 18 mg of compound 11 (yield: 96%).

The physicochemical properties of Compound 11 are as follows. ¹ H-NMR (270MHz, DMSO-d ₆ ) δ (ppm); 12.21 (1H, s), 10.39 (1
H, br), 8.60 (1H, br s), 8.09 (1H, s), 7.75 (2H, d, J = 8.4H
z), 7.57 (1H, d, J = 14.3Hz), 7.06 (1H, d, J = 14.3Hz), 6.99
(1H, d, J = 8.4Hz), 4.41 (3H, m), 4.12 (1H, m), 3.91 (4H,
m), 3.84 (3H, s), 3.81 (3H, s), 3.77 (1H, m), 3.69 (6H,
m), 2.68 (3H, s), 1.22 (1H, m), 1.11 (6H, d, J = 6.4Hz) IR (KBr) υ (cm ^-1 ); 1678,1643,1599,1515,1409,1251 , 12
12,1095,1032

Example 12 Synthesis of Compound 12 25 mg of Compound 2 obtained in Example 2 (0.06 mmo)
l) with 1.5 ml of acetonitrile and 48% hydrobromic acid
1.5 ml was added, and the mixture was stirred at room temperature for 1 hour. 1N Hydrobromic acid was added to this reaction mixture, and the mixture was extracted with chloroform. The chloroform layer was dried over anhydrous sodium sulfate and then concentrated under reduced pressure. The crude product obtained was treated with dichloromethane (2 m).
It was dissolved in 1, and phenylisocyanate was 0.033 at 0 ° C.
ml (0.30 mmol), then triethylamine (0.042 ml, 0.30 mmol) and added at 0 ° C. for 3
Stir for hours. To this reaction mixture was added 0.2 M acetate buffer of pH 4, and the mixture was extracted with chloroform. The chloroform layer was washed with saturated saline and dried over anhydrous sodium sulfate,
It was concentrated under reduced pressure. The obtained crude product was subjected to silica gel chromatography (20 ml, chloroform: methanol =
30: 1) and 18 mg of compound 12 (yield: 4
8%) was obtained.

The physicochemical properties of Compound 12 are as follows. ¹ H-NMR (270MHz, CDCl ₃ ) δ (ppm); 9.69 (1H, s), 8.35 (1H,
s), 7.78 (1H, d, J = 15.5Hz), 7.71 (1H, br s), 7.53 (2H, d, J
= 8.9Hz), 7.29 ~ 6.99 (5H, m), 6.91 (2H, d, J = 8.8Hz), 6.7
5 (1H, d, J = 15.6Hz), 4.44 (1H, m), 4.35 (1H, br d, J = 11.2H
z), 4.14 (1H, dd, J = 10.2,8.9Hz), 3.91 (3H, s), 3.85 (3H,
s), 3.69 (1H, br d, J = 10.2Hz), 3.09 (1H, dd, J = 9.8,9.8H
z), 2.52 (3H, s) FABMS (m / z); 620,618 (M + H) ⁺ IR (KBr) υ (cm ^-1 ); 1733,1699,1642,1603,1514,1444,14
12,1305,1253,1204,1092

Example 13 Synthesis of Compound 13 16 mg (0.062 mmol) of Compound 1 obtained in Example 1 was added to 0.1 mg of N, N-dimethylformamide in 3 mg (0.075 mmol) of 60% sodium hydride.
0.3 ml of N, N-dimethylformamide solution of
The mixture was stirred under an argon atmosphere at -20 ° C for 3 hours. 25 mg (0.076 mmol) of N, N- was added to the reaction solution of 4-nitrophenyl ester of 4-propyloxycinnamic acid.
0.5 ml of dimethylformamide solution was added, and the mixture was stirred at -20 ° C to 0 ° C for 1 hr. The reaction mixture was adjusted to pH 7
0.2M phosphate buffer was added, and the mixture was extracted with ethyl acetate. The ethyl acetate layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was purified by silica gel chromatography (20 ml, chloroform: methanol = 100: 1) to give compound 13 (22 m).
g (yield: 80%) was obtained.

The physicochemical properties of Compound 13 are as follows. ¹ H-NMR (400MHz, CDCl ₃ ) δ (ppm); 10.01 (1H, br), 7.77 (1H,
d, J = 15.4Hz), 7.32 ~ 6.93 (4H, m), 6.84 (1H, d, J = 15.7H
z), 6.67 (1H, br), 4.24 (1H, dd, J = 11.0,11.0Hz), 4.15 (1
H, dd, J = 11.0,4.7Hz), 3.94 (2H, t, J = 6.3Hz), 3.82 (3H,
s), 3.55 (1H, m), 2.59 (3H, s), 2.39 (1H, dd, J = 7.5,3.2H
z), 1.82 (2H, m), 1.31 (1H, dd, J = 4.9,3.7Hz), 1.05 (3H,
t, J = 7.4Hz) SIMS (m / z); 447 (M + H) ⁺ , 259 IR (KBr) υ (cm ^-1 ); 1697,1654,1596,1437,1388,1292,12
46,1213,1110

Example 14 Synthesis of Compound 14 16 mg (0.062 mmol) of Compound 1 obtained in Example 1 was obtained by adding 0.1 ml of N, N-dimethylformamide to 3 mg (0.075 mmol) of 60% sodium hydride.
0.3 ml of N, N-dimethylformamide solution of
The mixture was stirred under an argon atmosphere at -20 ° C for 3 hours. 25 mg (0.077 mmol) of N, N of 4-nitrophenyl ester of 4-propenyloxycinnamic acid was added to this reaction solution.
-Add 0.5 ml of dimethylformamide solution, and add -20 ° C.
The mixture was stirred at 0 ° C for 1 hour. PH 7 to this reaction mixture
0.2M phosphate buffer solution was added and extracted with ethyl acetate.
The ethyl acetate layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was subjected to silica gel chromatography (20 ml, chloroform:
The compound 14 was purified with methanol = 100: 1) to give 24
mg (yield: 87%) was obtained.

The physicochemical properties of Compound 14 are as follows. ¹ H-NMR (400MHz, CDCl ₃ ) δ (ppm); 10.51 (1H, br), 7.77 (1H,
d, J = 15.6Hz), 7.33 ~ 6.95 (4H, m), 6.84 (1H, d, J = 15.6H
z), 6.77 (1H, br), 6.07 (1H, m), 5.43 (1H, dd, J = 17.4,1.8
Hz), 5.31 (1H, dd, J = 10.5,1.2Hz), 4.57 (2H, dt, J = 5.1,1.
5Hz), 4.25 (1H, dd, J = 10.9,10.9Hz), 4.15 (1H, dd, J = 11.
0,4.6Hz), 3.82 (3H, s), 3.57 (1H, m), 2.60 (3H, s), 2.40
(1H, dd, J = 7.6,3.5Hz ), 1.32 (1H, dd, J = 4.7,3.7Hz) SIMS (m / z); 445 (M + H) +, 259 IR (KBr) υ (cm - ¹ ); 1701,1603,1486,1445,1388,1292,12
46,1215,1109

Example 15 Synthesis of Compound 15 To 3 mg (0.075 mmol) of 60% sodium hydride, 0.1 ml of N, N-dimethylformamide was added, and 16 mg (0.062 mmol) of Compound 1 obtained in Example 1 was added.
0.3 ml of N, N-dimethylformamide solution of
The mixture was stirred under an argon atmosphere at -20 ° C for 3 hours. 27 mg (0.076 mmol) of N, N- 4-pentyloxycinnamic acid 4-nitrophenyl ester was added to the reaction solution.
0.5 ml of dimethylformamide solution was added, and the mixture was stirred at -20 ° C to 0 ° C for 1 hr. The reaction mixture was adjusted to pH 7
0.2M phosphate buffer was added, and the mixture was extracted with ethyl acetate. The ethyl acetate layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was purified by silica gel chromatography (20 ml, chloroform: methanol = 100: 1) to give compound 15 (24 m).
g (yield: 82%) was obtained.

The physicochemical properties of Compound 15 are as follows. ¹ H-NMR (400MHz, CDCl ₃ ) δ (ppm); 10.34 (1H, br), 7.78 (1H,
d, J = 15.4Hz), 7.52 ~ 6.93 (4H, m), 6.85 (1H, d, J = 15.4H
z), 6.67 (1H, br), 4.24 (1H, dd, J = 11.0,11.0Hz), 4.15 (1
H, dd, J = 11.0,4.6Hz), 3.99 (2H, t, J = 6.6Hz), 3.82 (3H,
s), 3.55 (1H, m), 2.61 (3H, s), 2.39 (1H, dd, J = 7.6,3.4H
z), 1.81 (2H, m), 1.44 (4H, m), 1.31 (1H, dd, J = 4.9,3.7H
z), 0.94 (3H, t, J = 7.0Hz) SIMS (m / z); 475 (M + H) ⁺ , 259 IR (KBr) υ (cm ^-1 ); 1701,1628,1599,1457,1389 , 1255,12
16,1109

[0115]

INDUSTRIAL APPLICABILITY According to the present invention, compound (I) or a pharmacologically acceptable salt thereof has a high antitumor activity and is useful as an antitumor agent.

Claims (1)

[Claims] 1. The formula:{In the formula,IsOr [Chemical 4]And X is a hydrogen atom or CO₂CH₃And Z is
Represents Cl or Br, R is a hydrogen atom, CONR¹R² 
(In the formula, R¹And R² Is the same or different
Represents a child, a lower alkyl group or a phenyl group. ) Or(In the formula, n represents an integer of 0 to 4, R³ Is CH₂ ,, Oxygen atom, N-CH₃Or N-CH₂ CONR¹ R
 ²Represents R¹And R ² Is as defined above. ) Table
And W is a hydrogen atom or[In the formula, W¹ And W² Are the same or different, hydrogen
Atom or OR^Four(In the formula, R^Four Is a lower alkyl group or
Represents a lower alkenyl group. ) Represents. ]] Is represented. } Table
-89 derivative or its pharmacologically acceptable
salt.