JPH0565283A - Beta-carboline derivative - Google Patents

Abstract

PURPOSE:To provide the subject new compound having antitumor activity. CONSTITUTION:The compound of formula I [R<1> is group of formula II or III (n is 6 or 7)]. The compound of formula I can be produced by reacting a compound of formula IV with a compound of formula V in the presence of DPPA and triethylamine at room temperature.

Description

Detailed Description of the Invention

[0001]

The present invention relates to β having antitumor activity.
-Relating to carboline derivatives.

[0002]

2. Description of the Related Art Manzamine C having a chemical structure similar to that of the compound of the present invention is known to have antitumor activity (Tetrahedron Letters 28 , 5493, (1
987))

[0003]

DISCLOSURE OF THE INVENTION The present invention is directed to manzamine C.
By providing a similar compound, further excellent antitumor activity can be expected.

[0004]

The substance related to manzamine C according to the present invention is a compound represented by the following formula (1).

[Chemical 1] (Where R ¹ is

[Chemical 2] Or

[Chemical 3] And n is 6 or 7) The compound of the present invention is a compound of formula (2) (wherein R ² represents a lower alkyl group) and a compound of formula (3) (n is 6 or 7) Obtained by reacting the compound represented in the presence of DPPA and triethylamine at room temperature.

[Chemical 4]

[Chemical 5] The compounds of the formulas (2) and (3) used in this reaction are tetrahedron letter 30 , 6549 (1989).
Year).

[0005]

【Example】

Example 1 N- (1β-carbolinylacetyl) azacycloheptanamide 1-ethoxycarbonylmethyl-β-carboline (25
4 mg) was dissolved in ethanol (10 ml) and KOH0.
1g of 13g aqueous solution was added and stirred at room temperature for about 2 hours.
It was confirmed by LC (developing solvent: chloroform-methanol 9: 1) that the spot of the raw material disappeared and only the spot near the origin was left, and the reaction solution was concentrated to dryness under reduced pressure. DMF (5 ml) and azacycloheptane (0.11 ml) were added to the obtained residue, DPPA (0.71 ml) and then triethylamine (0.45 ml) were added, and the mixture was stirred at room temperature for about 1 ml.
Stir for 2 hours. R by TLC (developing solvent: ethyl acetate)
After confirming that no more spots around _f 0.3 were generated, the reaction solution was made alkaline with 10% NaOH aqueous solution and extracted with a mixed solvent of ethyl acetate-benzene (3: 1). The organic layer was washed with saturated brine, dried over Na ₂ SO ₄ , concentrated to dryness, and 680 mg of the resulting residue was separated using a silica gel column (Fuji-BW-300, 20 g, developed with ethyl acetate) to obtain 320 mg of the target compound. Was given. IR (KBr): 3600 to 3300 br, 2910,
1680, 1620, 1500, 1475, 1450,
1420, 1370, 1320, 1240, 1200,
800,740 cm ⁻¹ High resolution mass spectrum: Calculated value as C ₁₉ H ₂₁ N ₃ O 307.1686 Measured value 307.1681 PMR: 1.38 to 1.44 (m, 8H), 1.66
(M, 4H), 3.50 (t, 2H, J = 6.1H
z), 3.72 (t, 2H, J = 6.1 Hz), 7.2
6 (m, 1H), 7.56 (m, 2H), 7.86
(D, 1H, J = 5.2 Hz), 8.10 (d, 1H,
J = 7.7 Hz), 8.38 (d, 1H, J = 5.2H
z), 10.1 (s, 1H)

Example 2 N- (1β-carbolinylethyl) azacycloheptane N- (1β-carbolinylacetyl) azacycloheptaneamide (87.8 mg) was dissolved in THF (20 ml). LiAlH ₄ (90 mg) was added to this solution at room temperature and stirred for about 1 hour, and TLC (developing solvent: dichloromethane-ethyl acetate-methanol (7: 2: 1)) was used.
_A new spot appeared near _f 0.3, and the spot of the raw material disappeared. The reaction mixture was concentrated and the concentrated residue was converted into CH ₂
It was diluted with Cl ₂ (about 20 ml), 10% aqueous sodium hydroxide solution was added to decompose excess LiAlH ₄ , and the target product was extracted with CH ₂ Cl ₂ . The organic layer was dried over Na ₂ SO ₄ and concentrated to dryness. The concentrated residue (52 mg) was added to a silica gel column (F
uji-BW-300, 5 g, dichloromethane-ethyl acetate-methanol (developed with 7: 2: 1) was used to obtain a fraction containing the target substance, and this fraction was further added to an alumina column (Al.
₂ O ₃ , 20 g, developed with acetonitrile) to obtain 38 mg of the target substance. IR (KBr): 3700 to 3000 br, 745 cm
^-1 High resolution mass spectrum: Calculated value as C ₁₉ H ₂₃ N ₃ 293.1893, measured value 293.1900 PMR: 1.84 (m, 8H), 2.88 (m, 4)
H), 2.99 (t, 2H, J = 5.50 Hz), 3.
40 (t, 2H, J = 5.50 Hz), 7.26 (m,
1H), 7.55 (m, 2H), 7.84 (d, 1H,
J = 5.49 Hz), 8.13 (d, 1H, J = 7.8)
8 Hz), 8.27 (d, 1H, J = 5.31 Hz),
12.70 (s, 1H)

Example 3 N- (1β-carbolinylacetyl) azacyclooctanamide 1-ethoxycarbonylmethyl-β-carboline (45
0 mg) was dissolved in ethanol (10 ml) and KOH0.
1 ml of an aqueous solution containing 23 g was added, and the mixture was stirred at room temperature for about 2 hours, and TLC (developing solvent: dichloromethane-ethyl acetate-
It was confirmed that the spot of the raw material disappeared with methanol (7: 2: 1), leaving only the spot near the origin.
The reaction solution was concentrated to dryness under reduced pressure. DMF was added to the obtained residue.
(9 ml) azacyclooctane (200 mg) was added,
DPPA (0.83 ml) and then triethylamine (1.06 ml) were added, and the mixture was stirred at room temperature for about 12 hours. T
After confirming by LC (developing solvent: ethyl acetate) that a spot near R _f 0.3 was not formed any more, 10 was added to the reaction solution.
% NaOH aqueous solution to make it alkaline, ethyl acetate-
It was extracted with a mixed solvent of benzene (3: 1). The organic layer was washed with saturated brine, dried over Na ₂ SO ₄ and concentrated to dryness. The resulting residue (1.28 g) was purified by silica gel column (Fuji-).
After separation with BW-300 (50 g, developed with ethyl acetate), 505 mg of the target substance was obtained. IR (KBr): 3600 to 3000 br, 1620,
745 cm ⁻¹ High resolution mass spectrum: Calculated value as C ₂₀ H ₂₃ N ₃ O 321.1843 Measured value 321.1839 PMR: 1.25 (m, 2H), 1.37 (m, 2)
H), 1.43 (m, 2H), 1.71 (m, 2H),
1.75 (m, 2H), 3.44 (t, 2H, J = 6.
05Hz), 3.68 (t, 2H, J = 6.05H
z), 4.31 (s, 2H), 7.26 (m, 1H),
7.55 (m, 2H), 7.87 (d, 1H, J = 5.
50Hz), 8.09 (d, 1H, J = 7.69H
z), 8.33 (d, 1H, J = 5.22 Hz)

Example 4 N- (1β-carbolinylethyl) azacyclooctane N- (1β-carbolinylacetyl) azacyclooctaneamide (100 mg) was dissolved in THF (10 ml). LiAlH ₄ (100 mg) was added to this solution at room temperature, and the mixture was stirred for about 1 hour. Then, TLC (developing solvent: dichloromethane-ethyl acetate-methanol (7: 2: 1), R _f
A new spot appeared around 0.3, and the spot of the raw material disappeared. The reaction mixture was concentrated and the concentrated residue was converted to CH ₂ C.
l ₂ to decompose the excess LiAlH ₄ was added diluted 10% caustic soda aqueous solution (about 20 ml), the objective product was extracted with _CH 2 Cl _2. The organic layer was dried over Na ₂ SO ₄ and concentrated to dryness. The concentrated residue (84 mg) was passed through a silica gel column (Fu
ji-BW-300 (8 g, developed with dichloromethane-ethyl acetate-methanol (7: 2: 1)) to obtain a fraction containing the target compound, and this fraction was further added to an alumina column (Al ₂ O).
_3, 20 g, was obtained 63.1mg of the purified target substance by expansion in acetonitrile). IR (KBr): 3700 to 3000 br, 745c
m ⁻¹ high resolution mass spectrum: calculated as C ₂₀ H ₂₅ N ₃ 307.2051 measured 307.2037 PMR: 1.81 (m, 10H), 2.92 (m, 4)
H), 3.05 (t, 2H, J = 5.50 Hz), 3.
44 (t, 2H, J = 5.50 Hz), 7.26 (m,
1H), 7.53 (m, 2H), 7.84 (d, 1H,
J = 5.31 Hz), 8.12 (d, 1H, J = 7.8)
7 Hz), 8.28 (d, 1H, J = 5.31 Hz),
12.70 (s, 1H)

[0009]

The test results of the antitumor activity of the compounds of the present invention are shown below. Test method Cells used: P388, P388 / ADR, MKN-2
8, MKN-1, QG-56, PC-10, PC-1
4, C-1 medium used: RPMI1640 medium with fetal bovine serum 10%
And 2-hydroxymethyl disulf
The medium to which 10 μM of ide was added was used as a growth medium. Drug preparation: All samples were dissolved in DMSO to a concentration of 20 mg / ml. Then RPMI1640
Dilute with medium to test concentration range 100-0.41μg /
ml (common ratio 3). Culturing: After the addition of the drug, culturing was performed for 72 hours at 37 ° C. and 5% CO ₂ . Measurement: Viable cells were measured by the MTT assay method (Note 1). Judgment: The ratio of the number of surviving cells in the untreated control and the drug-added group was determined to obtain the cell survival rate (T / C%), and the 50% inhibitory concentration (IC ₅₀ ) was calculated from this. Note 1 MTT assay method According to the improved method of Mosmann's method. That is, 0.5% 3- (4,5-dimethyl) was added to the cell culture medium.
hiazol-2-yl) -2,5-diph-eny
ltetrazolium bromide (MTT)
The solution was added at 10 μ / well, reacted at 37 ° C. under 5% CO ₂ for 6 hours, and 0.01N HCl-10% sod was generated when MTT form-azan was produced.
Add the immodo-ecyl sulphate solution to 100
μl / well was added to dissolve MTTfor-mazan. After that, MTT form in each well
Absorbance of az-an is 577 nm as the main wavelength and 630 as the sub-wavelength.
The cell survival rate was determined by measuring with an ELISA analyzer (ETY-96, Toyo Sokki Co., Ltd.) in nm.

The results are shown in Table 1 below.

[Table 1] As is clear from the above table, the compounds of the present invention exhibit antitumor activity, and are particularly sensitive to adriamycin (doxorubicin) resistant cells (P388 / ADR), and to human-derived cells (MKN-28 to C-1). It is characterized in that the sensitivity is not significantly reduced as compared to the sensitivity to P388.

Claims (1)

[Claims] 1. A β-carboline derivative represented by the following formula (1): (Here, R ¹ is Or Represents, and n is 6 or 7)