JPH0525176A - Mitomycin derivative - Google Patents

Abstract

PURPOSE:To obtain a new compound having antitumor and antimicrobial activities. CONSTITUTION:A compound expressed by formula I [W is formula II (A is-N=, etc.; Q<1> and Q<2> are H, hydroxyl, amino, etc.); X is methoxy or amino; Y is H or methyl; Z is H, methyl or lower alkanoyl; either of R<1> and R<2> is carbamoyloxymethyl and the other is H or R<1> and R<2> together form methylenel, e.g. 1,6-demethyl-6-[(2-pyridylthio)methyl]mitomycin C. Furthermore, the compound expressed by formula I is obtained by reacting a compound expressed by formula III with a compound expressed by the formula WSH in the presence of a base such as triethylamine in an inert organic solvent such as THF at 0-30 deg.C and reacting the resultant compound expressed by formula IV with ammonia or ammonium acetate in an organic solvent such as methanol or with alcoholates of an alkali metal in a solvent such as methanol.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel mitomycin derivative having antitumor activity.

[0002]

BACKGROUND ART A mitomycin derivative modified with a methyl group at the 6-position related to the present invention is a mitomycin derivative in which a hydrogen atom of a methyl group is replaced with deuterium ( ² H) or tritium ( ³ H). 1-70490) and formula

[0003]

[Chemical 7]

[Wherein W'is RO or RS (wherein R is hydrogen,
A substituted or unsubstituted alkyl having 1 to 22 carbon atoms, a substituted or unsubstituted aralkyl, or a substituted or unsubstituted aryl)] is known (Japanese Patent Application Laid-Open No. 2-167282). ing.

[0005]

DISCLOSURE OF THE INVENTION The present invention is to provide a novel mitomycin derivative in which the methyl group at the 6-position is modified.

[0006]

The present invention provides formula (I)

[0007]

[Chemical 8]

{Where W is

[0009]

[Chemical 9]

[Wherein A represents -N = or -CR ³ = (in the formula, R ³ represents hydrogen, hydroxy, amino, aryl or lower alkyl), and Q ¹ and Q ² are the same or different. Represents hydrogen, hydroxy, amino, aryl or lower alkyl],

[0011]

[Chemical 10]

[Wherein A ¹ has the same meaning as A above, Q ³ has the same meaning as Q ¹ above, and B and D are the same or different.
Represents -N = or -CR ⁴ = (wherein R ⁴ has the same meaning as R ³ ) and E
Represents -O-, -S- or -NR ^5- , where R ⁵ has the same meaning as R ^3. ],

[0013]

[Chemical 11]

(Wherein A ² has the same meaning as A above, E ¹ has the same meaning as E above, and Q ⁴ has the same meaning as Q ¹ ),

[0015]

[Chemical 12]

(Wherein Q ⁵ , Q ⁶ , Q ⁷ and Q ⁸ are the same or different and are hydrogen, hydroxy, lower alkanoyloxy,
Hydroxymethyl or lower alkanoyloxymethyl) or

[0017]

[Chemical 13]

Wherein A ³ has the same meaning as A above, E ² has the same meaning as E above, and Q ⁹ has the same meaning as Q ¹ above, and X represents methoxy or amino. , Y represents hydrogen or methyl, Z represents hydrogen, methyl or lower alkanoyl, and R ¹ and R ^{2 each} represent carbamoyloxymethyl and the other represents hydrogen, or methylene (=
CH ₂ )}. Hereinafter, the compound represented by the formula (I) is referred to as the compound (I).
The same applies to compounds having other formula numbers.

In the definition of each group of formula (I), the lower alkyl group means a linear or branched alkyl group having 1 to 6 carbon atoms, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec- Butyl, tert-butyl, pentyl, neopentyl, hexyl and the like, and the alkanoyl moiety in the lower alkanoyl group and lower alkanoyloxy group and lower alkanoyloxymethyl group has 1 to 6 carbon atoms, for example, formyl, acetyl, propionyl, butyryl. , Isobutyryl, pivaloyl, valeryl and isovaleryl and the like, and aryl includes phenyl and naphthyl and the like.

Next, a method for producing the compound (I) will be described. Compound (I) can be produced according to the following steps.

[0021]

[Chemical 14]

(In the formula, n represents an integer of 2 or 3, and W, Y,
Z, R ¹ and R ² are as defined above) Step 1: Compound (III) is compound (II) and WSH (W is as defined above) in the presence of a base, if necessary. First, it can be obtained by reacting in a solvent inert to the reaction. Examples of the solvent used in the reaction include ether,
Tetrahydrofuran, dichloromethane, chloroform,
There are dimethylformamide (hereinafter referred to as DMF) and the like, which are used alone or in combination.

Examples of the base used in the reaction include organic bases such as pyridine and triethylamine.
The reaction is usually completed in 10 minutes to 24 hours at 0 to 30 ° C.
The starting compound (II) is a known compound described in JP-A 1-6275 and JP-A 1-70490. Step 2: Compound (Ia) [a compound (I) wherein X is NH ₂ ] can be obtained by reacting compound (III) with ammonia or ammonium acetate in a solvent inert to the reaction. it can.

The solvent used in the reaction may be any solvent which is inert to the reaction and dissolves the compound (III), and examples thereof include alcohols such as methanol and ethanol, ethers such as ether and tetrahydrofuran, dichloromethane and chloroform. Halogenated alkanes, acetonitrile,
DMF, dimethyl sulfoxide and the like can be used alone or in combination.

The reaction is usually carried out in the range of 0 to 30 ° C. for 1 hour to 14 hours.
It ends in days. Step 3: Compound (Ib) [a compound (I) wherein X is CH ₃ O] can be obtained by reacting compound (III) in methanol in the presence of a base. The base used in the reaction is an alkali metal or alkaline earth metal alcoholate, hydroxide, carbonate or bicarbonate, a tertiary amine or a quaternary ammonium hydroxide. The amount of base is in the range of 0.001 to 10 equivalents, preferably 0.01 to 3 equivalents, relative to compound (III).

The reaction is completed at 1 to 24 hours at 0 to 30 ° C.
In the compound (III) in which Z is represented by lower alkanoyl, the deacylation reaction also proceeds under the reaction conditions of steps 2 and 3, and the compound (Ia) obtained in steps 2 and 3 respectively
In and (Ib), a compound in which Z is a hydrogen atom can also be obtained.

The intermediates and target compounds in the above-mentioned production method are purified by a method commonly used in synthetic organic chemistry, for example,
It can be isolated and purified by subjecting to neutralization, filtration, extraction, washing, drying, concentration, recrystallization, various chromatography and the like. Further, the intermediate may be subjected to the next reaction without being particularly purified. Further, the compound (I) is
It may exist in the form of an adduct with water or various solvents, and these adducts are also included in the present invention.

Specific examples of the compound (I) obtained by the above production method are shown in Table 1.

[0029]

[Table 1]

[0030]

[Table 2]

* In the table, Ph represents phenyl and AcO represents acetoxy. Next, typical antitumor activity and acute toxicity test examples of compound (I) are shown. Test example 1. HeLaS ₃ cell growth inhibition test: HeLaS ₃ cells prepared at a concentration of 3 × 10 ⁴ cells / ml in MEM medium containing 10% fetal bovine serum and 2 mM glutamine were dispensed into each well of a 96-well microtiter plate in an amount of 0.1 ml.

After culturing overnight at 37 ° C. in a carbon dioxide gas incubator, a solution of the test compound appropriately diluted with the culture medium was added to 0.2.
Added 05 ml each. Cells in a carbon dioxide incubator
After culturing for 1 hour, the culture supernatant was removed, washed once with phosphate buffered saline, 0.1 ml of fresh medium was added to each well, and the cells were cultured at 37 ° C. for 72 hours in a carbon dioxide incubator. After removing the culture supernatant, 0.1 ml of a culture solution containing 0.02% neutral red was added to each well, and the cells were stained by culturing at 37 ° C. for 1 hour in a carbon dioxide gas incubator. After removing the culture supernatant, wash once with physiological saline and
The dye was extracted with N hydrochloric acid / 30% ethanol, and the amount of the extracted dye was measured as the absorbance at 550 nm using a microplate reader. Absorbance of untreated cells and test compounds
The growth inhibition rate of cells was calculated from the absorbance of cells treated with (various concentrations) according to the following formula.

[0033]

[Equation 1]

From the obtained growth inhibition rate, the cell growth was determined to be 50
The concentration of the test compound that caused% inhibition was calculated and used as the IC ₅₀ value. The results are shown in Table 2.

[0035]

[Table 3]

Test Example 2. Antitumor activity against Sarcoma 180 solid tumor: 5 × 10 ⁶ Sarcoma 180 cells were intraperitoneally transplanted into ddY mice, and cells were collected from ascites on day 7. After washing the collected cells once with sterile physiological saline,
The cells were suspended in sterile physiological saline to prepare a cell suspension of 5 × 10 ⁷ cells / ml.

0.1 ml of the obtained cell suspension was d with a body weight of 20 ± 2 g.
A solution of the test compound dissolved in physiological saline or a physiological saline containing polyoxyethylene sorbitan monolaurate was subcutaneously transplanted subcutaneously in the right axilla of a dY male mouse 24 hours after tumor implantation, and the solution was intravenously administered to 5 mice per group. 0.1 to 0.2 ml was administered.

[0038] 7 days after transplantation to measure the major axis of tumor (a) and minor axis (b), and tumor volume was calculated the value of a × b ^2/2. The antitumor activity of the test compound was represented by the ratio T / C of the tumor volume (C) of the control group to which the drug was not administered to the tumor volume (T) of the test compound administration group. The vertical axis represents T / C on a normal scale and the horizontal axis represents the dose on a logarithmic scale.T / C at each dose was plotted, and the relationship between dose and T / C was analyzed by the least squares method. ..
The dose showing T / C = 0.5 was calculated from the obtained regression line, and ED
₅₀ was calculated.

The results are shown in Table 3.

[0040]

[Table 4]

Test Example 3. Acute toxicity One test compound was intravenously administered once to 5 ddy mice per group, and life and death were observed for 14 days after the administration, and LD ₅₀ was calculated according to the Behrens-Kelver method from the mortality rate of each administration group. ..

The results are shown in Table 4.

[0043]

[Table 5]

The compound obtained by the present invention is useful as an antitumor agent and can be used as it is or in various dosage forms. For example, when the compound (I) is used as an injection, it is dissolved in a commonly used diluent in this field such as physiological saline, glucose injection, lactose injection, mannitol injection, or the like. It may be a freeze-dried injection or a powder injection mixed with sodium chloride according to the Pharmacopoeia. Further, polyethylene glycol, an auxiliary agent such as HCO-60 (surfactant; manufactured by Nikko Chemical Co., Ltd.) and the like, a carrier such as ethanol and / or liposomes and cyclodextrin may be contained. These injections are usually used for intravenous administration, but intraarterial administration, intraperitoneal administration, and intrathoracic administration are also possible.

Compound (I) and a suitable excipient, disintegrant,
Binders, lubricants and the like can be mixed and molded by a conventional method to give tablets, granules, powders, syrups and the like, which can be used as an oral preparation. Furthermore, compound (I) and a commonly used carrier can be mixed and molded by a conventional method to give a suppository, which can be rectally administered. The dose varies depending on the administration method, the type of compound (I), age, symptoms and the like, and the administration method can also be changed depending on the symptoms and dose. For example, it is possible to administer the dose in the range of 0.06 to 6 mg / kg once a week or once every three weeks.

Examples will be shown below.

[0047]

Example The physicochemical data of each compound were measured by the following instruments. ¹ H-NMR: JEOL JNM-GX270 (270MHz) Varian EM390 (90MHz) MS: JEOL JSM-D300 (measured by FAB method) IR: JEOL IR-810 (measured by KBr method) TLC: Silica gel Art5715 (Merck (Made by the company)

Example 1 6-demethyl-6-[(2-pyridylthio) methyl] mitomycin C (Compound 1) 1a-acetyl-7-demethoxy-6-demethyl-6,7-dihydro-7
-Ethylenedioxy-6-methylenemitomycin A (JP-A-1-70490, hereinafter referred to as compound α) (416 mg) was dissolved in dichloromethane (30 ml) to give 2-mercaptopyridine (112 mg).
Was added. After stirring the resulting solution at room temperature for 30 minutes, the reaction mixture was washed successively with saturated aqueous sodium hydrogen carbonate, phosphate buffer (pH 4) and saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and the desiccant was filtered off. After separation, the solvent was distilled off under reduced pressure.

The residue obtained is subjected to column chromatography.
Purify with (silica gel; chloroform: acetone = 1: 1),
A red fraction was obtained. After distilling off the solvent of this fraction, the obtained residue was dissolved again in a small amount of chloroform, and n-hexane was added to the solution to obtain a powdery substance. After the solvent was distilled off, it was dried sufficiently under vacuum to give 1a-acetyl-7-demethoxy-6-demethyl-6,7-dihydro-7-ethylenedioxy-6-[(2- 322 mg (61% yield) of pyridylthio) methyl] mitomycin A (compound a) was obtained.

Compound a (159 mg) was dissolved in methanol (50 ml), ammonium acetate (500 mg) was added, and the mixture was stirred at room temperature for 22 hours. The residue obtained by evaporating the solvent of the reaction mixture was purified by column chromatography (silica gel; chloroform: methanol = 30: 1 to 20: 1) to obtain a purple fraction.
By subjecting this fraction to the same operation as described above, 22.0 mg (yield 16%) of purple powdery compound 1 was obtained.

TLC: Rf = 0.31 (chloroform: methanol
= 9: 1) FAB-MS (m / z): 444 (M ⁺ +1); C ₂₀ H ₂₁ N ₅ O ₅ S = 443 IR (cm ^-1 ): 3420, 3300, 3200, 2930, 1720, 1610, 155
0, 1340, 1070 ¹ H-NMR: δ, ppm (270MHz, pyridine-d ₅ ) 2.12 (bs, 1H), 2.74 (bs, 1H), 3.12 (bs, 1H), 3.18 (s,
3H), 3.57 (bd, J = 13Hz, 1H), 4.01 (dd, J = 4.3 & 11.3
Hz, 1H), 4.50 (s, 2H), 4.56 (d, J = 12.8Hz, 1H), 5.09
(bt, J = 11Hz, 1H), 5.40 (dd, J = 4.3 & 10.3Hz, 1H),
6.91 (m, 1H), 7.2 ~ 7.25 (m, 1H), 7.37 (dt, J = 1.9 &
7.8Hz, 1H), 7.4 ~ 8.2 (br, 4H), 8.41 (bd, J = 4.2Hz,
1H)

Example 2 6-Demethyl-6-[[(3-hydroxypyridin-2-yl) thio] methyl] mitomycin C (Compound 2) 313 mg of compound α was dissolved in 30 ml of dichloromethane to give 3-hydroxy-2-mercapto. Pyridine (95 mg) and triethylamine (100 μl) were added, and the mixture was stirred at room temperature for 30 minutes. The reaction mixture was washed successively with phosphate buffer (pH 4), saturated aqueous sodium hydrogen carbonate solution and saturated brine, dried over anhydrous sodium sulfate, the desiccant was filtered off, and the solvent was evaporated under reduced pressure.

The obtained residue is subjected to column chromatography
Purification with (silica gel; chloroform: methanol = 20: 1) gave an orange fraction. By treating this fraction in the same manner as above, red powdery 1a-acetyl-7-demethoxy-6-demethyl-6,7-dihydro-7-ethylenedioxy-6-
190 mg (yield 47%) of [[(3-hydroxypyridin-2-yl) thio] methyl] mitomycin A (compound b) were obtained.

190 mg of compound b was added to anhydrous tetrahydrofuran 2
It was dissolved in 0 ml and allowed to stand at room temperature for 49 hours in a dry ammonia atmosphere. The residue obtained by distilling off the solvent of the reaction mixture was purified by column chromatography (silica gel; chloroform: methanol = 20: 1 to 10: 1) to obtain a purple fraction. By subjecting this fraction to the same treatment as above, 47.6 mg (yield 30%) of compound 2 in the form of a purple powder was obtained.

TLC: Rf = 0.19 (chloroform: methanol = 9: 1) FAB-MS (m / z): 460 (M ⁺ +1); C ₂₀ H ₂₁ N ₅ O ₆ S = 459 IR (cm ^-1 ): 3360, 3120, 2920, 2850, 1710, 1600, 155
0, 1430, 1370, 1330, 1280, 1260, 1210, 1060 ¹ H-NMR: δ, ppm (270MHz, pyridine-d ₅ ) 2.21 (bs, 1H), 2.79 (bs, 1H), 3.16 (bs, 1H ), 3.23 (s,
3H), 3.64 (d, J = 12.3Hz, 1H), 4.05 (dd, J = 4.3 & 11.
1Hz, 1H), 4.57 (d, J = 12.8Hz, 1H), 5.04 (bt, J = 10Hz,
1H), 5.41 (dd, J = 4.4 & 10.4Hz, 1H), 5.66 (d, J = 1
4.5Hz, 1H), 5.80 (d, J = 14.5Hz, 1H), 6.53 (dd, J = 6.8
& 7.7Hz, 1H), 7.00 (dd, J = 1.3 & 7.7Hz, 1H), 7.4 ~
8.0 (bs, 2H), 7.91 (dd, J = 1.1 & 6.6Hz, 1H), 8.6 ~ 9.
0 (bs, 2H), 9.33 (bs, 1H)

Example 3 6-Demethyl-6-[(2-pyrimidinylthio) methyl] mitomycin C (Compound 3) 420 mg of compound α was dissolved in dichloromethane, 111 mg of 2-mercaptopyrimidine and 50 μl of triethylamine were added, and the mixture was stirred at room temperature for 3 hours. It was stirred. The reaction mixture is phosphate buffered
(pH 4), washed successively with saturated saline and dried over anhydrous sodium sulfate, the desiccant was filtered off and the solvent was distilled off under reduced pressure. The obtained residue was purified by column chromatography (silica gel; chloroform: methanol = 25: 1) to obtain an orange fraction. By treating the obtained fraction in the same manner as in Example 1, red powdery 1a-acetyl-7-demethoxy-6-demethyl-6,7-dihydro-7-ethylenedioxy-6-
362 mg of [(2-pyrimidinylthio) methyl] mitomycin A (compound c) was obtained.

362 mg of compound c was added to anhydrous tetrahydrofuran 5
It was dissolved in 0 ml and allowed to stand at room temperature for 150 hours in a dry ammonia atmosphere. The residue obtained by evaporating the solvent of the reaction mixture was subjected to column chromatography (silica gel; chloroform:
Purification with methanol = 30: 1 to 15: 1) gave a purple fraction. The obtained fraction was treated in the same manner as in Example 1 to obtain 121 mg (yield 40%) of purple powdery compound 3.

TLC: Rf = 0.27 (chloroform: methanol = 9: 1) FAB-MS (m / z): 445 (M ⁺⁺ 1); C ₁₉ H ₂₀ N ₆ O ₅ S = 444 IR (cm ^-1 ): 3400, 3330, 3180, 2930, 1710, 1600, 157
0, 1550, 1460, 1440, 1380, 1340, 1200, 1070 ¹ H-NMR: δ, ppm (270MHz, pyridine-d ₅ ) 1.9 to 2.4 (bs, 1H), 2.74 (bs, 1H), 3.13 (bs , 1H), 3.1
9 (s, 3H), 3.60 (bd, J = 12Hz, 1H), 4.03 (dd, J = 4.2 &
11.2Hz, 1H), 4.53 (s, 2H), 4.56 (d, J = 12.8Hz, 1H),
5.08 (t, J = 10.4Hz, 1H), 5.42 (dd, J = 4.2 & 10.4Hz,
1H), 6.85 (t, J = 4.9Hz, 1H), 7.4 ~ 7.9 (bs, 2H), 7.4
~ 8.7 (br, 2H), 8.47 (d, J = 4.8Hz, 2H)

Example 4 6-Demethyl-6-[[(4,6-dimethylpyrimidin-2-yl) thio] methyl] mitomycin C
(Compound 4) 419 mg of compound α was dissolved in 30 ml of dichloromethane, 139 mg of 4,6-dimethyl-2-mercaptopyrimidine and 10 μl of triethylamine were added, and the mixture was stirred at room temperature for 2 hours and 15 minutes. The reaction mixture was directly subjected to column chromatography (silica gel, chloroform: methanol = 50: 1) for purification to obtain an orange fraction. By treating the obtained fraction in the same manner as in Example 1, red powdery 1a-acetyl-7-demethoxy-6-demethyl-6-[(4,6-dimethylpyrimidin-2-yl) thio] was obtained. 383 mg (69% yield) of methyl-6,7-dihydro-7-ethylenedioxymitomycin A (compound d) was obtained.

Compound d 383 mg was added to anhydrous tetrahydrofuran 5
It was dissolved in 0 ml and allowed to stand at room temperature for 48 hours in a dry ammonia atmosphere. The residue obtained by evaporating the solvent of the reaction mixture was purified by column chromatography (silica gel; chloroform: methanol = 30: 1 to 20: 1) to obtain a purple fraction. The obtained fraction was treated in the same manner as in Example 1 to obtain 200 mg of Compound 4 in the form of purple powder (yield 62%).

TLC: Rf = 0.28 (chloroform: methanol = 9: 1) FAB-MS (m / z): 473 (M ⁺⁺ 1); C ₂₁ H ₂₄ N ₆ O ₅ S = 472 IR (cm ^-1 ): 3380, 3200, 2950, 1720, 1610, 1580, 155
0, 1440, 1340, 1270, 1230, 1070 ¹ H-NMR: δ, ppm (270MHz, pyridine-d ₅ ) 2.13 (bs, 1H), 2.26 (s, 6H), 2.74 (bs, 1H), 3.14 ( bs,
1H), 3.18 (s, 3H), 3.57 (bd, J = 12.6Hz, 1H), 4.01 (d
d, J = 4.2 & 11.2Hz, 1H), 4.49 (s, 2H), 4.56 (d, J = 1
2.8Hz, 1H), 5.09 (bt, J = 10.8Hz, 1H), 5.40 (dd, J =
4.3 & 10.4Hz, 1H), 6.52 (s, 1H), 7.4 ~ 7.9 (bs, H), 7.
9 ~ 8.8 (br, 2H)

Example 5 6-Demethyl-6-[(2-imidazolylthio) methyl] mitomycin C (Compound 5) 422 mg of compound α was dissolved in 40 ml of dichloromethane, and 105 mg of 2-mercaptoimidazole and 50 ml of triethylamine.
Was added, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was washed with saturated brine, dried over anhydrous sodium sulfate, the desiccant was filtered off, and the solvent was evaporated under reduced pressure. The obtained residue was purified by column chromatography (silica gel; chloroform: methanol = 20: 1 to 10: 1) to obtain a yellow fraction.
By treating the obtained fraction in the same manner as in Example 1, 1a-acetyl-7-demethoxy-6-demethyl-6,7-dihydro-7-ethylenedioxy-6-[(2 -Imidazolylthio) methyl] mitomycin A (Compound e) 146mg
(Yield 28%) was obtained.

103 mg of compound e was dissolved in anhydrous tetrahydrofuran 2
It was dissolved in 0 ml and allowed to stand at room temperature for 191 hours in a dry ammonia atmosphere. The residue obtained by evaporating the solvent of the reaction mixture was subjected to column chromatography (silica gel; chloroform:
Purification with methanol = 9: 1) gave a brown fraction. By treating the obtained fraction in the same manner as in Example 1, 60.6 mg (yield 70%) of a brown powdery compound 5 was obtained.

TLC: Rf = 0.20 (chloroform: methanol = 9: 1) FAB-MS (m / z): 433 (M ⁺⁺ 1); C ₁₈ H ₂₀ N ₆ O ₅ S = 432 IR (cm ^-1 ): 3370, 3200, 3020, 2940, 1720, 1710, 161
0, 1570, 1550, 1540, 1460, 1340, 1070 ¹ H-NMR: δ, ppm (270MHz, pyridine-d ₅ ) 2.09 (bs, 1H), 2.78 (dd, J = 1.8 & 4.3Hz, 1H), 3.14
(d, J = 4.4Hz, 1H), 3.21 (s, 3H), 3.65 (dd, J = 1.8 &
12.7Hz, 1H), 4.00 (dd, J = 4.3 & 11.1Hz, 1H), 4.60
(d, J = 12.7Hz, 1H), 5.03 (bt, J = 11Hz, 1H), 5.18 (d,
J = 14.4Hz, 1H), 5.30 (d, J = 14.4Hz, 1H), 5.37 (dd, J
= 4.4 & 10.5Hz, 1H), 6.70 (d, J = 1.7Hz, 1H), 7.14 (d,
J = 2.2Hz, 1H), 7.3 ~ 7.8 (br, 2H), 8.40 (bs, 1H),
9.19 (bs, 1H), 14.0 (bs, 1H)

Example 6 6-demethyl-6-[[(1-methylimidazol-2-yl) thio] methyl] mitomycin C (Compound 6) 331 mg of compound α was dissolved in 40 ml of dichloromethane to give 2-mercapto-1-methyl. Imidazole 94.6 mg and triethylamine 100 μl were added, and the mixture was stirred at room temperature for 3 hours and 15 minutes. The reaction mixture was washed successively with phosphate buffer (pH 4) and saturated saline and dried over anhydrous sodium sulfate, then the desiccant was filtered off and the solvent was distilled off under reduced pressure. The obtained residue was subjected to column chromatography (silica gel; chloroform: methanol = 3
Purification at 0: 1) gave a yellow fraction. By treating the obtained fraction in the same manner as in Example 1, 1a-acetyl-7-demethoxy-6-demethyl-6,7-dihydro-7-ethylenedioxy-6-[[(( 224 mg of 1-methylimidazol-2-yl) thio] methyl] mitomycin A (Compound f) (yield 5
8%) obtained.

207 mg of compound f was mixed with anhydrous tetrahydrofuran 3
It was dissolved in 0 ml and left standing at room temperature for 232 hours in a dry ammonia atmosphere. The residue obtained by evaporating the solvent of the reaction mixture was subjected to column chromatography (silica gel; chloroform:
Purification with methanol = 30: 1) gave a brown fraction. By treating the obtained fraction in the same manner as in Example 1, 89.0 mg (yield 51%) of brown powdery compound 6 was obtained.

TLC: Rf = 0.32 (chloroform: methanol = 9: 1) FAB-MS (m / z): 448 (M ⁺ +2); C ₁₉ H ₂₂ N ₆ O ₅ S = 446 IR (cm ^-1 ): 3370, 3180, 2930, 1720, 1710, 1660, 160
0, 1560, 1550, 1450, 1440, 1400, 1350, 1330, 1220,
1060 ¹ H-NMR: δ, ppm (90MHz, pyridine-d ₅ ) 2.04 (s, 1H), 2.75 (bs, 1H), 3.10 (bs, 1H), 3.18 (s, 3
H), 3.48 (s, 3H), 3.58 (dd, J = 2 & 12Hz, 1H), 3.94 (d
d, J = 4 & 10Hz, 1H), 4.53 (d, J = 12Hz, 1H), 4.96 (b
t, J = 10Hz, 1H), 5.11 (bs, 2H), 5.32 (dd, J = 4 & 10H
z, 1H), 6.76 (d, J = 2Hz, 1H), 6.99 (d, J = 2Hz, 1H),
7.3 ~ 7.6 (bs, 2H), 8.1 ~ 8.5 (bs, 1H), 8.5 ~ 9.0 (bs, 1
H)

Example 7 6-Demethyl-6-[[(1,2,4-triazol-3-yl) thio] methyl] mitomycin C (Compound 7) Compound α 420 mg was dissolved in dichloromethane 40 ml to give 3-mercapto- 110 mg of 1,2,4-triazole was added, and the mixture was stirred at room temperature for 4 hours. The reaction mixture was washed with saturated brine and dried over anhydrous sodium sulfate, the desiccant was filtered off, and the solvent was evaporated under reduced pressure. The obtained residue was purified by column chromatography (silica gel; chloroform: methanol = 30: 1) to obtain a reddish purple fraction. By treating the obtained fraction in the same manner as in Example 1, red-purple powdery 1a-acetyl-7-demethoxy-6-demethyl-6,7-dihydro-7-ethylenedioxy-6-[[[ (1,2,4-triazol-3-yl) thio]
Methyl] mitomycin A (compound g) 77.2 mg (15% yield)
Obtained.

52.2 mg of compound g was added to anhydrous tetrahydrofuran
Dissolve in 20 ml, 30 hours at room temperature in a dry ammonia atmosphere for 30 hours
Let stand for a minute. The residue obtained by distilling off the solvent of the reaction mixture was purified by column chromatography (silica gel; chloroform: methanol = 9: 1) to obtain a purple fraction. By treating the obtained fraction in the same manner as in Example 1,
1.5 mg (3.5% yield) of purple powdery compound 7 was obtained.

TLC: Rf = 0.16 (chloroform: methanol
= 9: 1) FAB-MS (m / z): 434 (M ⁺ +1); C ₁₇ H ₁₉ N ₇ O ₅ S = 433 IR (cm ^-1 ): 3370, 3200, 3000, 2930, 1720, 1710, 166
0, 1600, 1560, 1550, 1480, 1450, 1340, 1210, 1070 ¹ H-NMR: δ, ppm (270MHz, pyridine-d ₅ ) 2.15 (bs, 1H), 2.79 (bs, 1H), 3.15 (bs , 1H), 3.22 (s,
3H), 3.63 (bd, J = 12.8Hz, 1H), 4.01 (dd, J = 4.2 & 1
1.2Hz, 1H), 4.55 (d, J = 12.6Hz, 1H), 5.06 (bt, J = 10
Hz, 1H), 5.09 (d, J = 14.5Hz, 1H), 5.17 (d, J = 14.5H
z, 1H), 5.38 (dd, J = 4.4 & 10.4Hz, 1H), 7.4 ~ 7.8 (b
s, 4H), 8.51 (s, 1H), 8.6 ~ 8.9 (br, 1H)

Example 8. 6-Demethyl-6-[[(4-methyl-1,
2,4-Triazol-3-yl) thio] methyl] mitomycin C (Compound 8) Compound a (421 mg) was dissolved in dichloromethane (40 ml) to prepare 3-mercapto-4-methyl-1,2,4-triazole (116 mg) and triethylamine (100 μ). In addition, the mixture was stirred at room temperature for 1 hour and 45 minutes. The reaction mixture was washed successively with a phosphate buffer (pH 4) and saturated brine, dried over anhydrous sodium sulfate, the desiccant was filtered off, and the solvent was evaporated under reduced pressure. The obtained residue was purified by column chromatography (silica gel; chloroform: methanol = 30: 1) to obtain a yellow fraction. By treating the obtained fraction in the same manner as in Example 1, 1a-acetyl-7-demethoxy-6-demethyl-6,7-dihydro-7-ethylenedioxy-6-[[(( 196 mg (yield 37%) of 4-methyl-1,2,4-triazol-3-yl) thio] methyl] mitomycin A (compound h) was obtained.

154 mg of compound h was added to anhydrous tetrahydrofuran 4
It was dissolved in 0 ml and allowed to stand at room temperature for 327 hours under a dry ammonia atmosphere. The residue obtained by evaporating the solvent of the reaction mixture was subjected to column chromatography (silica gel; chloroform:
Purification with methanol = 30: 1) gave a purple fraction. The obtained fraction was treated in the same manner as in Example 1 to obtain 91.0 mg (yield 70%) of Compound 8 in the form of purple powder.

TLC: Rf = 0.41 (chloroform: methanol = 9: 1) FAB-MS (m / z): 449 (M ⁺ +2); C ₁₈ H ₂₁ N ₇ O ₅ S = 447 IR (cm ^-1 ): 3350, 3130, 2920, 2850, 1720, 1710, 160
0, 1570, 1550, 1540, 1450, 1340, 1210, 1060 ¹ H-NMR: δ, ppm (90MHz, pyridine-d ₅ ) 1.96 (bs, 1H), 2.60 (m, 1H), 3.01 (m, 1H ), 3.06 (s, 3
H), 3.38 (s, 3H), 3.47 (bd, J = 12Hz, 1H), 3.87 (dd, J
= 4 & 11Hz, 1H), 4.46 (d, J = 12Hz, 1H), 4.90 (bt, J =
11Hz, 1H), 5.13 (dd, J = 4 & 11Hz, 1H), 5.37 (s, 2
H), 7.48 (bs, 2H), 8.13 (bs, 2H), 8.28 (s, 1H)

Example 9 6-Demethyl-6-[[(1-methyltetrazol-5-yl) thio] methyl] mitomycin C (Compound 9) 1a-Acetyl-7-demethoxy-6-demethyl-6,7-dihydro -7
418 mg of -ethylenedioxy-6-methylenemitomycin A (JP-A-1-70490) was dissolved in 40 ml of dichloromethane, 115 mg of 5-mercapto-1-methyltetrazole and 100 μl of triethylamine were added, and the mixture was stirred at room temperature for 1 hour.
The reaction mixture was washed successively with a phosphate buffer (pH 4) and saturated brine, dried over anhydrous sodium sulfate, the desiccant was filtered off, and the solvent was evaporated under reduced pressure. The obtained residue was purified by column chromatography (silica gel; chloroform: methanol = 30: 1) to obtain an orange fraction. By treating the obtained fraction in the same manner as in Example 1, 1a-acetyl-7-demethoxy-6-demethyl-6,7-dihydro-7-ethylenedioxy-6-[[(( 1-methyltetrazole-
5-yl) thio] methyl] mitomycin A (compound i)
7 mg (yield 55%) was obtained.

277 mg of compound i was added to anhydrous tetrahydrofuran 4
It was dissolved in 0 ml and allowed to stand at room temperature for 107 hours in a dry ammonia atmosphere. The residue obtained by distilling off the solvent of the reaction mixture was used for preparative TLC (silica gel; chloroform: methanol = 9: 1).
And purified to give a purple fraction. The obtained fraction was solvent-extracted, and the obtained extract was treated in the same manner as in Example 1 to obtain 57.5 mg (yield 25%) of purple powdered compound 9.

TLC: Rf = 0.27 (chloroform: methanol
= 9: 1) FAB-MS (m / z): 437 (M ⁺ +1); C ₁₆ H ₂₀ N ₈ O ₅ S = 436 IR (cm ^-1 ): 3350, 3300, 3200, 2940, 1710, 1600, 156
0, 1550, 1450, 1340, 1070 ¹ H-NMR: δ, ppm (90MHz, pyridine-d ₅ ) 2.0 (bs, 1H), 2.63 (m, 1H), 3.03 (m, 1H), 3.10 (s, 3
H), 3.49 (bd, J = 13Hz, 1H), 3.65 (s, 3H), 3.90 (dd, J =
5 & 11Hz, 1H), 4.46 (d, J = 13Hz, 1H), 4.93 (bt, J =
11Hz, 1H), 5.22 (dd, J = 5 & 10Hz, 1H), 5.34 (s, 2
H), 7.3 ~ 7.6 (br, 2H), 8.0 ~ 8.5 (br, 2H)

Example 10.6-Demethyl-6-[[(1-phenyltetrazol-5-yl) thio] methyl] mitomycin C
(Compound 10) 420 mg of compound α was dissolved in 40 ml of dichloromethane, 179 mg of 5-mercapto-1-phenyltetrazole and 100 μl of triethylamine were added, and the mixture was stirred at room temperature for 4 hours and 10 minutes. The reaction mixture was washed successively with a phosphate buffer (pH 4) and saturated brine, dried over anhydrous sodium sulfate, the desiccant was filtered off, and the solvent was distilled off. The obtained residue was subjected to column chromatography (silica gel; chloroform: methanol = 30: 1).
To 20: 1) to give a yellow fraction. By treating the obtained fraction in the same manner as in Example 1, yellow powdery 1a-acetyl-7-demethoxy-6-demethyl-6,7-dihydro was obtained.
-7-Ethylenedioxy-6-[[(1-phenyltetrazole-
5-yl) thio] methyl] mitomycin A (compound j)
0 mg (yield 53%) was obtained.

Compound j 304 mg was added to anhydrous tetrahydrofuran 4
It was dissolved in 0 ml and allowed to stand at room temperature for 18 hours in a dry ammonia atmosphere. The residue obtained by distilling off the solvent of the reaction mixture was used for preparative TLC (silica gel; chloroform: methanol = 9: 1).
And purified to give a purple fraction. The obtained fraction was subjected to solvent extraction, and the obtained extract was treated in the same manner as in Example 1 to obtain 11.9 mg (yield 5%) of purple powdery compound 10.

TLC: Rf = 0.42 (chloroform: methanol = 9: 1) FAB-MS (m / z): 512 (M ⁺ +2); C ₂₂ H ₂₂ N ₈ O ₅ S = 510 IR (cm ^-1 ): 3400, 3200, 2930, 1720, 1710, 1600, 156
0, 1550, 1450, 1340, 1070 ¹ H-NMR: δ, ppm (90MHz, pyridine-d ₅ ) 2.00 (bs, 1H), 2.56 (m, 1H), 3.04 (m, 1H), 3.12 (s, 3
H), 3.52 (dd, J = 2 & 13Hz, 1H), 3.93 (dd, J = 5 & 11H
z, 1H), 4.48 (d, J = 13Hz, 1H), 4.96 (t, J = 11Hz, 1
H), 5.30 (dd, J = 5 & 11Hz, 1H), 5.47 (s, 2H), 7.2 ~
8.1 (m, 7H), 8.28 (bs, 2H)

Example 11 1. Demethyl-6-[(2-thiazolinylthio) methyl] mitomycin C (Compound 11) Compound α (415 mg) was dissolved in dichloromethane (30 ml), 2-mercaptothiazoline (120 mg) and triethylamine (50 μl) were added, and the mixture was stirred at room temperature for 45 minutes. It was stirred. The reaction mixture was washed successively with saturated aqueous sodium hydrogen carbonate, phosphate buffer (pH 4) and saturated brine, dried over anhydrous sodium sulfate, the desiccant was filtered off, and the solvent was evaporated. The obtained residue is subjected to column chromatography (silica gel; chloroform: methanol = 30: 1 to 20: 1).
Purification was carried out to give an orange fraction. The obtained fraction was used in Example 1
1a-acetyl-7-demethoxy-6-demethyl-6,7-dihydro-7-ethylenedioxy-6-[(2-thiazolinylthio) methyl] mitomycin A (compound 336 mg (yield 63%) of k) was obtained.

336 mg of compound k was added to anhydrous tetrahydrofuran 3
It was dissolved in 0 ml and allowed to stand at room temperature for 116 hours in a dry ammonia atmosphere. The residue obtained by evaporating the solvent of the reaction mixture was subjected to column chromatography (silica gel; chloroform:
Purification with methanol = 20: 1) gave a purple fraction. The obtained fraction was treated in the same manner as in Example 1 to obtain 188 mg (yield 67%) of Compound 11 in the form of purple powder.

[0082] TLC: Rf = 0.29 (chloroform: methanol = 9: 1) FAB-MS (m / z): 452 (M + +1); C 18 H 21 N 5 O 5 S 2 = 451 IR (cm - ¹ ): 3380, 3200, 2940, 1720, 1710, 1600, 157
0, 1550, 1540, 1450, 1340, 1320, 1220, 1060 ¹ H-NMR: δ, ppm (270MHz, pyridine-d ₅ ) 2.21 (bs, 1H), 2.81 (bs, 1H), 3.04 (t, J = 8.0Hz, 2H),
3.15 (bs, 1H), 3.25 (s, 3H), 3.64 (bd, J = 12.8Hz, 1
H), 3.96 (t, J = 8.2Hz, 2H), 4.03 (dd, J = 4.2 & 11.2H
z, 1H), 4.58 (d, J = 12.8Hz, 1H), 4.81 (d, J = 14.3Hz,
1H), 5.09 (bt, J = 11Hz, 1H), 5.13 (d, J = about 14Hz, 1
H), 5.39 (dd, J = 4.1 & 10.5Hz, 1H), 7.4 ~ 7.9 (bs, 2
H), 7.8 ~ 8.0 (bs, 1H), 8.4 ~ 8.6 (bs, 1H)

Example 12 6. Demethyl-6-[(2-thiazolylthio) methyl] mitomycin C (Compound 12) 416 mg of compound α was dissolved in 30 ml of dichloromethane, 116 mg of 2-mercaptothiazole and 50 μl of triethylamine were added, and the mixture was stirred at room temperature for 2 hours. Stir for 10 minutes. The reaction mixture was directly purified by column chromatography (silica gel; chloroform: methanol = 50: 1 to 20: 1) to obtain an orange fraction. By treating the obtained fraction in the same manner as in Example 1, 1a-acetyl-7-demethoxy-6-demethyl-6,7-dihydro-7-ethylenedioxy-6-[(2 -Thiazolylthio) methyl] mitomycin A (compound m) 335m
g (yield 63%) was obtained.

335 mg of compound m was added to anhydrous tetrahydrofuran 5
It was dissolved in 0 ml and allowed to stand at room temperature for 72 hours in a dry ammonia atmosphere. The residue obtained by evaporating the solvent of the reaction mixture was purified by column chromatography (silica gel; chloroform: methanol = 30: 1 to 20: 1) to obtain a purple fraction. The obtained fraction was treated in the same manner as in Example 1 to obtain 177 mg of purple powdery compound 12 (yield 63%).

TLC: Rf = 0.31 (chloroform: methanol
= 9: 1) FAB-MS (m / z): 450 (M ⁺ +1); C ₁₈ H ₁₉ N ₅ O ₅ S ₂ = 449 IR (cm ^-1 ): 3370, 3200, 2930, 1720, 1600 , 1570, 155
0, 1450, 1340, 1210, 1060 ¹ H-NMR: δ, ppm (270MHz, pyridine-d ₅ ) 2.20 (bs, 1H), 2.80 (bs, 1H), 3.16 (bs, 1H), 3.23 (s,
3H), 3.63 (d, J = 12.5Hz, 1H), 4.02 (dd, J = 4.2 & 11.
2Hz, 1H), 4.56 (d, J = 12.8Hz, 1H), 5.08 (bt, J = 11Hz,
1H), 5.18 (d, J = 14.3Hz, 1H), 5.30 (d, J = 14.5Hz, 1
H), 5.38 (dd, J = 4.4 & 10.4Hz, 1H), 6.78 (d, J = 4.6H
z, 1H), 7.4 ~ 7.9 (bs, 2H), 7.51 (d, J = 4.6Hz, 1H),
8.4 ~ 8.7 (bs, 2H)

Example 13 6. Demethyl-6-[(2-benzothiazolylthio) methyl] mitomycin C (Compound 13) 263 mg of compound α was dissolved in 30 ml of dichloromethane, and 105 mg of 2-mercaptobenzothiazole and triethylamine 1
00 μl was added, and the mixture was stirred at room temperature for 4 hours. The reaction mixture was washed successively with a phosphate buffer (pH 4) and a saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, the desiccant was filtered off, and the solvent was evaporated under reduced pressure. The obtained residue was subjected to column chromatography (silica gel; chloroform: methanol =
30: 1) to give a yellow fraction. The obtained fraction was treated in the same manner as in Example 1 to give a yellow powder of 1a-acetyl-6-[(2-benzothiazolylthio) methyl-7-demethoxy-6-demethyl-6,7-dihydro. 198 mg (yield 54%) of -7-ethylenedioxymitomycin A (compound n) was obtained.

198 mg of compound n was added to anhydrous tetrahydrofuran 2
It was dissolved in 0 ml and allowed to stand in a dry ammonia atmosphere for 41 hours. The residue obtained by evaporating the solvent of the reaction mixture was purified by column chromatography (silica gel; chloroform: methanol = 30: 1) to obtain a purple fraction. The obtained fraction was treated in the same manner as in Example 1 to obtain 61.3 mg (yield 36%) of purple powdered compound 13.

TLC: Rf = 0.40 (chloroform: methanol
= 9: 1) FAB-MS (m / z): 500 (M ⁺ +1); C ₂₂ H ₂₁ N ₅ O ₅ S ₂ = 499 IR (cm ^-1 ): 3450, 3370, 3150, 2950, 1720 , 1600, 156
0, 1550, 1540, 1460, 1330, 1060 ¹ H-NMR: δ, ppm (90MHz, pyridine-d ₅ ) 2.19 (bs, 1H), 2.80 (bs, 1H), 3.16 (bs, 1H), 3.21 ( s,
3H), 3.70 (bd, J = 12.3Hz, 1H), 4.00 (dd, J = 4.2 & 1
1.2Hz, 1H), 4.66 (d, J = 12.8Hz, 1H), 5.04 (bt, J = 12
Hz, 1H), 5.36 (dd, J = 4.0 & 10.4Hz, 1H), 5.59 (d, J =
15.0Hz, 1H), 5.71 (d, J = 15.0Hz, 1H), 7.15 ~ 7.25 (m,
1H), 7.4 ~ 7.9 (br, 3H), 7.5 ~ 7.6 (m, 2H), 8.3 ~ 8.5
(bs, 1H), 8.62 (m, 1H)

Example 14.6 Demethyl-6-[[(5-methyl-1,
3,4-Thiadiazol-2-yl) thio] methyl] mitomycin C (Compound 14) Compound α 430 mg was dissolved in dichloromethane 40 ml, and 2-mercapto-5-methyl-1,3,4-thiadiazole 136 mg and triethylamine 100 μl were dissolved. In addition, the mixture was stirred at room temperature for 1 hour and 50 minutes. The reaction mixture was washed successively with a phosphate buffer (pH 4) and saturated brine, dried over anhydrous sodium sulfate, the desiccant was filtered off, and the solvent was evaporated under reduced pressure. The obtained residue was purified by column chromatography (silica gel; chloroform: methanol = 30: 1) to obtain an orange fraction. By treating the obtained fraction in the same manner as in Example 1, 1a-acetyl-7-demethoxy-6-demethyl-6,7-dihydro-7-ethylenedioxy-6-[[(( 448 mg (yield 79%) of 5-methyl-1,3,4-thiazol-2-yl) thio] methyl] mitomycin A (compound p) was obtained.

428 mg of compound p was added to anhydrous tetrahydrofuran 4
It was dissolved in 0 ml and left to stand under a dry ammonia atmosphere for 63 hours. The residue obtained by evaporating the solvent of the reaction mixture was purified by column chromatography (silica gel; chloroform: methanol = 30: 1) to obtain a purple fraction. By treating the obtained fraction in the same manner as in Example 1, 176 mg (yield 49%) of purple powdered Compound 14 was obtained.

[0091] TLC: Rf = 0.28 (chloroform: methanol = 9: 1) FAB-MS (m / z): 465 (M + +1); C 18 H 20 N 6 O 5 S 2 = 464 IR (cm - ¹ ): 3400, 3300, 3200, 2920, 1720, 1600, 156
0, 1550, 1340, 1210, 1060 ¹ H-NMR: δ, ppm (270MHz, pyridine-d ₅ ) 2.12 (bs, 1H),
2.12 (s, 3H), 2.72 (bs, 1H), 3.13 (bs, 1H), 3.15 (s, 3
H), 3.56 (bd, J = 12.9Hz, 1H), 4.00 (dd, J = 4.3 & 11.
1Hz, 1H), 4.56 (d, J = 12.7Hz, 1H), 5.05 (bt, J = 11H
z, 1H), 5.37 (dd, J = 4.2 & 10.5Hz, 1H), 5.53 (s, 2H),
7.4 ~ 7.8 (br, 2H), 7.9 ~ 8.5 (br, 2H)

Example 15 5. Demethyl-6-[(2-benzoxazolylthio) methyl] mitomycin C (Compound 15) Compound α (422 mg) was dissolved in dichloromethane (30 ml), and 2-mercaptobenzoxazole (157 mg) and triethylamine (50 μl) were added. The mixture was stirred at room temperature for 1 hour and 40 minutes. The reaction mixture was washed successively with a phosphate buffer (pH 4) and saturated brine, dried over anhydrous sodium sulfate, the desiccant was filtered off, and the solvent was evaporated under reduced pressure. The obtained residue is subjected to column chromatography (silica gel; chloroform: methanol = 30: 1).
And purified to give a yellow fraction. The obtained fraction was used in Example 1
By the same treatment as that of 1a-acetyl-6-[(2-benzoyloxazolylthio) methyl] -7-
Demethoxy-6-demethyl-6,7-dihydro-7-ethylenedioxymitomycin A (compound r) was obtained at 393 mg (68% yield).

373 mg of compound r was added to anhydrous tetrahydrofuran 3
It was dissolved in 0 ml and left for 22 hours under a dry ammonia atmosphere. The residue obtained by distilling off the solvent of the reaction mixture was purified by column chromatography (silica gel; chloroform: methanol = 30: 1 to 20: 1) to obtain a purple fraction.
By treating the obtained fraction in the same manner as in Example 1, 166 mg (yield 53%) of compound 15 in the form of purple powder was obtained.

TLC: Rf = 0.35 (chloroform: methanol = 9: 1) FAB-MS (m / z): 484 (M ⁺⁺ 1); C ₂₂ H ₂₁ N ₅ O ₆ S = 483 IR (cm ^-1 ): 3450, 3370, 3310, 3180, 2930, 1730, 172
0, 1600, 1570, 1550, 1480, 1450, 1390, 1340, 1070 ¹ H-NMR: δ, ppm (270MHz, Pyridine-d ₅ ) 2.14 (bs, 1H), 2.78 (bs, 1H), 3.14 (bs , 1H), 3.21 (s,
3H), 3.67 (bd, J = 12.8Hz, 1H), 4.01 (dd, J = 4.3 & 1
1.1Hz, 1H), 4.61 (d, J = 12.6Hz, 1H), 5.04 (bt, J = 9.
7Hz, 1H), 5.28 (bs, 2H), 5.37 (dd, J = 4.1 & 10.5Hz,
1H), 7.06 to 7.17 (m, 2H), 7.29 to 7.32 (m, 1H), 7.4 to 7.
7 (bs, 2H), 7.81 ~ 7.85 (m, 1H), 8.1 ~ 8.4 (bs, 1H),
8.5 ~ 8.9 (bs, 1H)

Example 16 6.-Demethyl-6-[[(2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) thio] methyl] mitomycin C (Compound 16) Compound α 215 mg was added to dichloromethane 20 ml. Dissolved in 1-thio
195 mg of -β-D-glucose tetraacetate and 50 μl of triethylamine were added, and the mixture was stirred at room temperature for 3 hours and 50 minutes. The reaction mixture was washed successively with a phosphate buffer (pH 4) and saturated brine, dried over anhydrous sodium sulfate, the desiccant was filtered off, and the solvent was evaporated under reduced pressure. The obtained residue was purified by column chromatography (silica gel; chloroform: methanol = 30: 1) to obtain a yellow fraction. By treating the obtained fraction in the same manner as in Example 1, 1a-acetyl-7-demethoxy-6,7-dihydro-7-ethylenedioxy-6-demethyl-6-[[(( 182 mg (yield 45%) of 2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) thio] methyl] mitomycin A (compound s) was obtained.

182 mg of compound s was mixed with anhydrous tetrahydrofuran 3
It was dissolved in 0 ml and allowed to stand in a dry ammonia atmosphere for 187 hours. The residue obtained by evaporating the solvent of the reaction mixture was purified by column chromatography (silica gel; chloroform: methanol = 30: 1) to obtain a purple fraction. The obtained fraction was treated in the same manner as in Example 1 to obtain 98.1 mg (yield 60%) of purple powdery compound 16.

TLC: Rf = 0.36 (chloroform: methanol = 9: 1) FAB-MS (m / z): 697 (M ⁺⁺ 1); C ₂₉ H ₃₆ N ₄ O ₁₄ S = 696 IR (cm ^-1 ): 3430, 3330, 3200, 2940, 1750, 1600, 155
0, 1430, 1370, 1340, 1230, 1040 ¹ H-NMR: δ, ppm (270MHz, pyridine-d ₅ ) 1.9 to 2.2 (bs, 1H), 1.99 (s, 3H × 2), 2.01 (s, 3H × 2),
2.75 (bs, 1H), 3.12 (bs, 1H), 3.24 (s, 3H), 3.60 (d, J
= 12.8Hz, 1H), 4.02 (d, J = 13.9Hz, 1H), 4.02 (dd, J =
3.9 & 10.8Hz, 1H), 4.13 (ddd, J = 2.6, 4.8 & 10.1Hz,
1H), 4.22 (d, J = 13.2Hz, 1H), 4.40 (dd, J = 2.3 & 12.
4Hz, 1H), 4.58 (dd, J = 4.9 & 12.6Hz, 1H), 4.59 (d, J
= 12.8Hz, 1H), 5.03 (m, 1H), 5.23 (d, J = 10.1Hz, 1
H), 5.38 (dd, J = 4.3 & 10.5Hz, 1H), 5.52 (t, J = 9.5H
z, 1H), 5.58 (t, J = 9.5Hz, 1H), 5.74 (t, J = 9.2Hz, 1
H), 7.3 ~ 7.8 (bs, 2H), 7.95 (bs, 2H)

Example 17.6-Demethyl-6-[(β-D-glucopyranosylthio) methyl] mitomycin C (Compound 17) Compound 16 (24.7 mg) obtained in Example 16 was added to a solution of ammonia in methanol (6.1 It was dissolved in N), the reaction vessel was sealed, and the mixture was allowed to stand at room temperature for 10 hours and 30 minutes. The solvent of the reaction mixture was distilled off, and reverse phase short column chromatography (Bond Elute
C18; water to water-acetonitrile, 60:40) to give a purple fraction. The organic solvent of this fraction was distilled off under reduced pressure,
The obtained aqueous solution was freeze-dried to obtain 14.8 mg (yield 79%) of purple amorphous compound 17.

FAB-MS (m / z): 530 (M ⁺ +2); C ₂₁ H ₂₈ N ₄ O ₁₀ S =
528 IR (cm ^-1 ): 3400, 3320, 2920, 1710, 1600, 1560, 155
0, 1530, 1450, 1340, 1070 ¹ H-NMR: δ, ppm (270MHz, pyridine-d ₅ ) [main peak] 2.01 (bs, 1H), 2.70 (bs, 1H), 3.09 (b
s, 1H), 3.14 (s, 3H), 3.55 (bd, J = 12Hz, 1H), 3.9 ~ 4.
2 (m, 6H), 4.21 (d, J = 13.4Hz, 1H), 4.29 (dd, 6.8, 11.
5Hz, 1H), 4.53 (d, J = 12.6Hz, 1H), 4.60 (bd, J = 10.1
Hz, 1H), 5.04 (bt, J = 11Hz, 1H), 5.11 (d, J = 9.2Hz,
1H), 5.33 (dd, J = 4.0 & 10.4Hz, 1H), 6.60 (bs, 1H),
7.11 (bs, 1H), 7.3 ~ 7.7 (bs, 2H), 7.78 (bs, 2H)

Example 18.6-Demethyl-6-[(2-thienylthio) methyl] mitomycin C (Compound 18) 1a-Acetyl-7-demethoxy-6-demethyl-6,7-dihydro-7
430 mg of -ethylenedioxy-6-methylenemitomycin A (JP-A-1-70490) was dissolved in 40 ml of dichloromethane, 100 μl of 2-mercaptothiophene and 100 μl of triethylamine were added, and the mixture was stirred at room temperature for 1 hour and 20 minutes.

The reaction mixture was washed successively with a phosphate buffer (pH 4) and saturated saline, dried over anhydrous sodium sulfate, filtered by a desiccant and the solvent was distilled off under reduced pressure. The obtained residue was subjected to column chromatography (silica gel; chloroform:
Purification with methanol = 30: 1) gave a red fraction. By treating the obtained fraction in the same manner as in Example 1, red powdery 1a-acetyl-7-demethoxy-6-demethyl-6,7-
189 mg (yield 34%) of dihydro-7-ethylenedioxy-6-[(2-thienylthio) methyl] mitomycin A (compound t) was obtained.

176 mg of the compound t obtained above was dissolved in 40 ml of anhydrous tetrahydrofuran, and the mixture was allowed to stand at room temperature for 163 hours in a dry ammonia atmosphere. The residue obtained by evaporating the solvent of the reaction mixture is purified by column chromatography (silica gel; chloroform: methanol = 20: 1), and further purified by preparative TLC (chloroform: methanol = 9: 1). Gave a purple fraction. The obtained fraction was solvent-extracted, and the obtained extract was treated in the same manner as in Example 1 to obtain 47.5 mg (yield 32%) of Compound 18 in the form of purple powder.

TLC: Rf = 0.40 (chloroform: methanol
= 1: 1) FAB-MS (m / z): 449 (M ⁺ +1); C ₁₉ H ₂₀ N ₄ O ₅ S ₂ = 448 IR (cm ^-1 ): 3400, 3320, 3270, 3200, 1720 , 1710, 160
0, 1560, 1550, 1440, 1340, 1220, 1070 ¹ H-NMR: δ, ppm (270MHz, pyridine-d ₅ ) 1.9 ~ 2.2 (br, 1H), 2.72 (bs, 1H), 3.12 (bs, 1H ), 3.1
9 (s, 3H), 3.54 (bd, J = about 13Hz, 1H), 4.01 (dd, J = 4.3
& 11.1Hz, 1H), 4.25 (s, 2H), 4.42 (d, J = 12.7Hz, 1
H), 5.02 (bt, J = about 11Hz, 1H), 5.39 (dd, J = 4.3 & 10.
4Hz, 1H), 6.91 (dd, J = 3.7 & 5.4Hz, 1H), 7.15 ~ 7.25
(m, overlap with pyridine, 1H), 7.38 (dd, J = 1.2 & 5.4Hz, 1
H), 7.4 ~ 7.7 (br, 2H), 8.0 ~ 8.3 (br, 2H)

[0104]

INDUSTRIAL APPLICABILITY The present invention provides a novel mitomycin derivative having excellent antitumor activity.

Claims (1)

What is claimed is: (Claim 1) Formula (I) {In the formula, W is [Chemical formula 2] [In the formula, A represents -N = or -CR ³ = (in the formula, R ³ represents hydrogen, hydroxy, amino, aryl or lower alkyl), and Q ¹ and Q ² are the same or different and each represents hydrogen or hydroxy. , Amino, aryl or lower alkyl], embedded image [Wherein A ¹ has the same meaning as A described above, Q ³ has the same meaning as Q ¹ described above, and B and D are the same or different and -N = or -CR
⁴ = (wherein R ⁴ has the same meaning as R ³ ) and E represents -O-, -S- or -NR ^5- (wherein R ⁵ has the same meaning as R ³ )] , (In the formula, A ² has the same meaning as A, E ¹ has the same meaning as E, and Q ⁴ has the same meaning as Q ^1. ) (Wherein Q ⁵ , Q ⁶ , Q ⁷ and Q ⁸ are the same or different and represent hydrogen, hydroxy, lower alkanoyloxy, hydroxymethyl or lower alkanoyloxymethyl) or (Wherein A ³ has the same meaning as A above, E ² has the same meaning as E above, Q ⁹ has the same meaning as Q ¹ above), X represents methoxy or amino, and Y Represents hydrogen or methyl, Z
Represents hydrogen, methyl or lower alkanoyl, and R ¹ and R ^{2 each} represent carbamoyloxymethyl and the other represents hydrogen, or they together represent methylene (= CH ₂ )}.