JPH0119386B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention provides novel derivatives of fredericamycin A, more specifically, the following general formula (): [In the formula, R represents a group -COR ₁ (wherein R ₁ represents an alkyl group, a lower alkyl group, a lower alkoxy group, or a phenyl group that may be substituted with a halogen atom)] A fredericamycin A-diacyl derivative represented by Regarding. Conventionally, fredericamycin A (FCRC-48), in which R in the formula (
A (NSC-305263)) is known to be isolated [J. Antibiotcs, Vol. 34, pp. 1389-1401 (1981) and Vol. 34, pp. 1402-1407 (1981)]. However, this fredericamycin A has weak antibacterial activity and is unstable. Therefore, in order to overcome this drawback of fredericamycin A, the present inventor synthesized various derivatives and examined their pharmacological action and stability, and found that the diacyl derivative represented by the above formula () was excellent. The present invention was completed based on the discovery that it has antibacterial and antitumor effects and is extremely stable compared to fredericamycin A. Accordingly, the present invention provides fredericamycin A-diacylated derivatives useful as antibacterial and anticancer agents. The acyl group of the diacyl derivative of the present invention (-
COR ₁ ) is one in which R ₁ is represented by an alkyl group or a phenyl group which may have a substituent, and the alkyl group preferably has 1 to 18 carbon atoms, and the substituent for the phenyl group is Examples thereof include linear or branched lower alkyl groups having 1 to 6 carbon atoms, lower alkoxy groups having 1 to 6 carbon atoms, and halogen atoms such as chlorine, bromine, fluorine, and iodine. The fredericamycin A-diacyl derivative of the present invention can be obtained by converting fredericamycin A into a carboxylic acid represented by R ₁ -COOH (where R ₁ is the same as above) or a reaction thereof. It is produced by reacting sexual derivatives. As the reactive derivatives, acid halides, acid anhydrides, mixed acid anhydrides, active esters, etc. are used, and in this case, 3 to 10% of fredericamycin A is used.
It is preferable to carry out the reaction using two molar equivalents of the acylating agent in a solvent such as pyridine at a temperature of 0 to 4°C for 2 to 48 hours. Alternatively, a condensing agent such as DCC can be used to directly react with the carboxylic acid. The results of testing the antibacterial activity, antitumor activity, and stability of the representative compound of the present invention thus obtained are as follows. (1) Antibacterial activity The minimum inhibitory concentration (MIC) of fredericamycin A-diacetate (compound 1) against various microorganisms is shown in Table 1 in comparison with fredericamycin A. Furthermore, fredericamycin A
The MIC for any microorganism is 100μg/ml
That's all. Test bacteria culture conditions: Inoculum size 10 ⁶ cells/ml. For bacteria, use Mueller Hinton Agar (manufactured by Difco) at 37°C for 18
The cells were cultured for ~20 hours, and in the case of yeast and mold, they were cultured for 120 hours at 28°C in a glucose-peptone medium.

【table】

[Table] Table 2 shows the MICs of various fredericamycin A-diacyl derivatives. The culture conditions for the test bacteria were the same.

[Table] (2) Antitumor effects of the compounds of the present invention and fredericamycin A on Ehrlich carcinoma (Ehrlich) and Meth-
The therapeutic effect on A (ascites type) was tested by the following method. The results are shown in Table 3. The survival effects in the table are expressed as a percentage of the ratio of survival days (T) in the treated group to survival days (C) in the untreated group. Experimental method: (i) Ehrlich 5 × 10 ⁶ tumor cells were implanted intraperitoneally into ICR mice (♀, CLEA Japan), and 24 hours later, the test compound was intraperitoneally administered once a day for a total of 10 times. did. (ii) Meth-A (ascites type) 1×10 ⁶ tumor cells were implanted into the peritoneal cavity of CDF ₁ mice (Charles River, Japan), and after 24 hours, the test compound was administered once a day for a total of 10 times. It was administered intraperitoneally.

【table】

[Table] (3) Stability The stability of Compound 5 of the present invention and Fredericamycin A in an aqueous solution was tested by the following method.
The results are shown in Table 4. Experimental method: Dissolve the test compound in dimethyl sulfoxide,
The final concentration of the test compound was adjusted to 10 μg/ml by diluting with physiological saline. For this test liquid,
The residual rate of the test compound after a predetermined time was measured by high performance liquid chromatography (HPLC method).

[Table] Next, examples will be given and explained. Example 1 0.54 g (1.0 mmol) of fredericamycin A was dissolved in 20 ml of pyridine, and 5 ml of pyridine was added under stirring at 0°C.
1.02 g (10 mmol) of acetic anhydride dissolved in was added dropwise over about 30 minutes, and the mixture was stirred at 0° C. for 3 hours. The reaction solution was poured into 200 ml of ice-cooled 2N hydrochloric acid, and extracted with ethyl acetate. The ethyl acetate layer was washed successively with dilute hydrochloric acid and water, and dried over anhydrous sodium sulfate. After evaporation, ethyl acetate was distilled off under reduced pressure, and the residue was recrystallized from a mixture of ethyl acetate and acetic acid to obtain 0.52 g (yield: 83.5%) of yellowish brown crystals of fredericamycin A-diacetate (compound 1). Melting point 300℃ or higher Mass M ⁺ m/z623 UV λ ^EtOH _nax (ε) 393nm (21200), 374nm (32100), 359nm
(27200), 333nm (22400), 319nm (21400),
305nm (17300), 258nm (49800), 235nm
(46600) IR ν ^KBr _nax cm ^-1 1780, 1720, 1690, 1655, 1625
(Figure 1) PMR δppm (CDCl ₃ ) 12.02 (1H, br.s), 10.32 (1H, br), 3.84
(3H, s), 3.21 (2H, t), 2.45 (6H, s),
1.56 (3H, d) (Figure 2) Example 2 Dissolve 0.54 g (1.0 mmol) of Fredericamycin A in 20 ml of pyridine, and add 5 ml of pyridine while stirring at 0°C.
0.60 g of isovaleric acid chloride dissolved in
(5.0 mmol) was added dropwise over about 30 minutes, and
Stir for hours. The reaction solution was poured into 200 ml of ice-cooled 2N hydrochloric acid, and extracted with ethyl acetate. The ethyl acetate layer was washed successively with dilute hydrochloric acid and water, and dried over anhydrous sodium sulfate. After the filtration, ethyl acetate was distilled off under reduced pressure, and 100 ml of ether was added to the residue to collect the precipitate.
Recrystallization from an ethyl acetate-acetic acid-methanol mixture gave 0.35 g (yield 49.5%) of yellowish brown crystals of fredericamycin A-diisovalate (compound 2). Melting point 260℃ MaSS M ⁺ m/z707 UV λ ^EtOH _nax (ε) 393nm (23800), 374nm (33700), 359nm
(28500), 333nm (23500), 319nm (22800),
305nm (19900), 260nm (50600), 235nm
(44000) IR ν ^KBr _nax cm ^-1 1775, 1720, 1690, 1655, 1620
(Figure 3) PMR δppm (CDCl ₃ ) 12.04 (1H, br.s), 9.82 (1H, br), 3.83 (3H,
s), 3.22 (2H, t), 2.63 (4H, t), 1.60
(3H, d), 1.09 (12H, d) (Figure 4) Example 3 0.54 g (1.0 mmol) of fredericamycin A was dissolved in 25 ml of pyridine, and 2.54 g of p-methylbenzoic anhydride ( 10 mmol) little by little,
After stirring at 0°C for 5 hours, the mixture was further left at 0°C for 1 day. Pour the reaction solution into 200ml of ice-cooled 2N hydrochloric acid,
The deposited precipitate was collected by filtration, washed with water, and dried. After washing this precipitate twice with 100 ml of hot isopropyl ether, it was recrystallized from a mixture of ethyl acetate, acetic acid, and methanol.
Yellowish brown crystals of methyl benzoate (compound 3)
0.61 g (yield 78.7%) was obtained. Melting point 300℃ or higher MaSS M ⁺ m/z775 UV λ ^EtOH _nax (ε) 393nm (23200), 374nm (34200), 359nm
(28800), 333nm (23700), 319nm (22800),
305nm (18900), 254nm (78300) IR ν ^KBr _nax cm ^-1 1750, 1720, 1690, 1660, 1625
(Fig. 5) PMR δppm (CDCl ₃ ) 12.02 (1H, br), 9.47 (1H, br), 8.12 (4H,
d), 7.27 (4H, d), 3.80 (3H, s), 3.17
(2H, t), 2.48 (2H, t), 2.39 (6H, s),
1.68 (3H, d) (Figure 6) Examples 4-11 The following compounds were obtained in the same manner as in Examples 1-3.
The compound is represented by R in the formula (). Compound 4 Melting point 300℃ or higher, yellowish brown crystal MaSS M ⁺ m/z651 UV λ ^EtOH _nax 393nm (22100), 374nm (31700), 359nm
(26200), 333nm (21900), 319nm (21200),
305nm (17700), 258nm (51200), 235nm
(46400) IR ν ^KBr _nax cm ^-1 1775, 1720, 1690, 1655, 1620
(Figure 7) PMR δppm (CDCl ₃ ) 12.01 (1H, br.s), 9.87 (1H, br), 3.87 (3H,
s), 3.25 (2H, t), 2.81 (4H, q), 2.49
(2H, t), 1.70 (3H, d), 1.33 (6H, t) (Figure 8) Compound 5 Melting point 162-164℃, yellowish brown crystal MaSS M ⁺ m/z735 UV λ ^EtOH _nax (ε) 393nm (23300), 374nm (34900), 359nm
(29200), 333nm (24200), 319nm (23300),
305nm (19000), 258nm (53800), 235nm
(50100) IR ν ^KBr _nax cm ^-1 1775, 1720, 1690, 1655, 1620
(Figure 9) PMR δppm (CDCl ₃ ) 12.04 (1H, s), 10.06 (1H, br.s), 3.86
(3H, s), 3.22 (2H, t), 2.77 (4H, t),
2.47 (2H, t), 1.59 (3H, d), 0.90 (6H,
t) (Figure 10) Compound 6 Melting point 120-122℃, yellowish brown crystal MaSS M ⁺ m/z847 UV λ ^EtOH _nax (ε) 393nm (23600), 374nm (35000), 359nm
(29300), 333nm (24100), 319nm (22800),
305nm (18500), 260nm (54000), 235nm
(50200) IR ν ^KBr _nax cm ^-1 1775, 1720, 1690, 1655, 1620
(Fig. 11) PMR δppm (CDCl ₃ ) 12.00 (1H, br.s), 10.27 (1H, br.s), 3.86
(3H, s), 3.21 (2H, t), 2.77 (4H, t),
2.47 (2H, t), 1.51 (3H, d), 0.84 (6H,
t) (Figure 12) Compound 7 Melting point 112-114℃, yellowish brown crystal MaSS M ⁺ m/z903 UV λ ^EtOH _nax (ε) 393nm (23100), 374nm (34100), 359nm
(28600), 333nm (23500), 319nm (22200),
305nm (18200), 260nm (53500), 235nm
(49500) IR ν ^KBr _nax cm ^-1 1775, 1720, 1690, 1655, 1625
(Fig. 13) PMR δppm (CDCl ₃ ) 12.02 (1H, br,) 10.16 (1H, br), 3.85 (3H,
s), 3.22 (2H, t), 2.76 (4H, t), 2.47
(2H, t), 1.58 (3H, d), 0.86 (6H, t) (Figure 14) Compound 8 Melting point 300℃ or higher, yellowish brown crystal MaSS M ⁺ m/z747 UV λ ^EtOH _nax (ε) 393nm (24000), 374nm (35400), 359nm
(29700), 333nm (24700), 319nm (24100),
305nm (20000), 255nm (64900), 237nm
(70000) IR ν ^KBr _nax cm ^-1 1750, 1720, 1690, 1655, 1625
(Fig. 15) PMR δppm (CDCl ₃ ) 12.08 (1H, br), 9.23 (1H, br), 3.81 (3H,
s), 3.19 (2H, t), 2.49 (2H, t), 1.73
(3H, d) (Figure 16) Compound 9 Melting point 300℃ or higher, yellowish brown crystal MaSS M ⁺ m/z831 UV λ ^EtOH _nax (ε) 393nm (23800), 374nm (35100), 359nm
(29500), 333nm (24600), 319nm (23600),
305nm (17800), 254nm (81700) IR ν ^KBr _nax cm ^-1 1750, 1720, 1690, 1660, 1625
(Fig. 17) PMR δppm (CDCl ₃ ) 12.08 (1H, br), 9.66 (1H, br), 8.16 (4H,
d), 7.32 (4H, d), 3.80 (3H, s), 3.16
(2H, t), 2.95 (2H, m), 2.48 (2H, t),
1.66 (3H, d), 1.26 (12H, d) (Figure 18) Compound 10 Melting point 300℃ or higher, yellowish brown crystal MaSS M ⁺ m/z807 UV λ ^EtOH _nax (ε) 393nm (23100), 374nm (33600), 359nm
(28300), 333nm (23600), 319nm (22800),
304nm (20700), 263nm (85700) IR ν ^KBr _nax cm ^-1 1740, 1720, 1690, 1655, 1620, 1605 PMR δppm (CDCl ₃ ) 12.04 (1H, br), 9.86 (1H, br), 8.19 (4H ,
d), 6.95 (4H, d), 3.82 (6H, s), 3.79
(3H, s), 3.15 (2H, t), 2.47 (2H, t),
1.61 (3H, d) (Figure 20) Compound 11 Melting point 300℃ or higher, yellowish brown crystal MaSS M ⁺ m/z815, 817, 819 UV λ ^EtOH _nax (ε) 393nm (24300), 374nm (36200), 359nm
(30800), 333nm (25800), 319nm (24900),
305nm (20800), 258nm (87200) IR ν ^KBr _nax cm ^-1 1750, 1720, 1690, 1655, 1625
(Figure 21) PMR δppm (CDCl ₃ ) 12.10 (1H, br), 9.40 (1H, br), 8.17 (4H,
d), 7.45 (4H, d), 3.82 (3H, s), 3.17
(2H, t), 2.48 (2H, t), 1.69 (3H, d)
(Figure 22)

[Brief explanation of drawings]

Figures 1 and 2 show the IR and PMR of compound 1.
It is a drawing showing a spectrum. FIG. 3 and FIG. 4 are drawings showing the IR and PMR spectra of Compound 2. Figures 5 and 6 show the IR of compound 3.
and a drawing showing a PMR spectrum. FIG. 7 and FIG. 8 are drawings showing the IR and PMR spectra of Compound 4. FIG. 9 and FIG. 10 are drawings showing the IR and PMR spectra of Compound 5. FIG. 11 and FIG. 12 are drawings showing the IR and PMR spectra of Compound 6. FIG. 13 and FIG. 14 are drawings showing the IR and PMR spectra of Compound 7. FIG. 15 and FIG. 16 are drawings showing the IR and PMR spectra of Compound 8. Figures 17 and 18 show the results of compound 9.
It is a drawing showing IR and PMR spectra. 1st
9 and 20 are drawings showing the IR and PMR spectra of Compound 10. Figures 21 and 22
The figure shows the IR and PMR spectra of Compound 11.

Claims (1)

[Claims] First-order general formula () [In the formula, R represents a group -COR ₁ (wherein R ₁ represents an alkyl group, a lower alkyl group, a lower alkoxy group, or a phenyl group that may be substituted with a halogen atom)] A fredericamycin A-diacyl derivative represented by .