JPS59175493A - Mitomycin derivative and its preparation - Google Patents

Abstract

NEW MATERIAL:A compound shown by the formula I {X is group shown by the formula II, group shown by the formula R3-S-S-CH2CH2-[R3 is 1-7C alkyl, cycloalkyl, -(CH2)nCO2R4 (n is 1-3; R4 is methyl or ethyl), pyridyl, 1-3C alkanoylmino-substituted phenyl, group shown by the formula III]; R1 and R2 are H, methyl but not H simultaneously; wavy line is alpha, beta bond; when carbamoyloxy methyl at 9-position is alpha, R3 is amino group-substituted 1-7C alkyl, or not group shown by the formula III, and when carbamoyloxymethyl at 9-position is beta and R1 is methyl, R3 is not H}. EXAMPLE:7-N, 7'-N-Dithiodiethylene dimitomycin D. USE:An antimicrobial agent. PREPARATION:A compound shown by the formula V is reacted with a compound shown by the formula R3'-SH[R3' is 1-7C alkyl, cycloalkyl, -(CH2)nCO2R4, or 1-3C alkanoylamino-substituted phenyl].

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to useful new mitomycin derivatives and methods for their production.

In more detail, the present invention is based on the general formula (1) [wherein R3 is a linear or branched alkyl or cycloalkyl having 1 to 7 carbon atoms (the alkyl or cycloalkyl is a hydroxyl or cycloalkyl having 1 to 3 carbon atoms] may be substituted with amino acids),
-(,C!H2)nO02R4(n is 1, 2 or 3, R
4 is methyl or ethyl), pyridyl, carbon number 1-
Phenyl substituted with alkanoylamino of 3, Suar.

RI and R2 are the same or different and are hydrogen or methyl, but not hydrogen at the same time.

W represents an α or β bond.

Regardless of the above definition, when carbamyloxymethyl at the 9-position is α, R3 has 1 to 7 carbon atoms substituted with an amine group.
A mitomycin derivative (hereinafter referred to as compound (1
). The same applies to compounds with other formula numbers) and their production methods.

Mitomycins are generally known as antibiotics having antibacterial and antitumor activities. Typical mitomycins include mitomycin A, B, O, porphyromycin (Merck Index 9th edition), mitomycin D5E (Japanese Patent Application Laid-Open No. 122797/1982), mitomycin F (% open 55-45322), etc. can be given. These mitomycins have the following chemical structures and can be obtained by culturing Streptomyces kespitosas strains.

X), 9 to 8 to 10 R),
RBM 1 tomy C1rl, A 0(
jH31II II II C! H31 (tt
B 00 horse 1 HC horsettONH
2 cloudy old + m0H3H p D NH2□ H0H3s
E NH2-m 0H3C! H3tt
F 0OH3mm+ OH30yen Porfiromycin NH2++um
OH3CH3 Also, various derivatives of these mitomycins are known. Among these derivatives, compounds in which the amino group at the 7-position of mitomycin C is substituted with the following groups are known: 2-thiazole amino, full 7-lylamino, 2-
Propenylamino, 2-propynylamino, 2-thenylmethylamino, carbamoylmethylamino [US Pat. No. 4,268,676, J, Me4. -Qhem,,
Embryo, 975 (1981)]. Furthermore] Japanese Patent Publication No. 57-18
859 is a mitomycin derivative in which the amino group at position 7 is substituted with various heterocycles such as quinolinyl, pyrazolyl, or thiazole skeleton; a mitomycin derivative in which position 7 is substituted with a nitrogen-containing heterocycle such as 1-pyrrolinyl or 1-indolinyl; 2-Mercaptoethylamino (Example 11S, 33) or 2-(ethylthio)ethylamino (Example 32)
Mitomycin derivatives substituted with and the like have been disclosed.

However, in all of the above documents, the 7-position amino group is -5
There is no disclosure of mitomycin derivatives substituted with groups having an -S- bond.

As a result of research on new mitomycin derivatives, 7
A compound having a substituent having a -5-S-bond at the position (1
) was found to have excellent antibacterial activity.

Next, the present invention will be explained in more detail.

In the definition of R3 in general formula (1), a straight chain or branched alkyl or cycloalkyl having 1 to 7 carbon atoms (the alkyl or cycloalkyl may be substituted with 1 to 3 hydroxyl or 1 amine) ) as methyl, ethyl,
n-propyl, i-propyl, n-butyl, B-butyl, t-butyl, cyclopentyl, cyclohexyl, 2
-Hydroxyethyl, 3-hydroxypropyl, 2.3
-Dihydroxypropyl, 2-aminoethyl, 6-aminopropyl, 4-aminobutyl, 4-hydroxycyclohexyl, etc. 1, in the definition of R3 -(OH,)
nc 02R4 is ethoxycarbonylmethyl,
Examples of the pyridyl include 2-pyridyl and 4-pyridyl, and examples of the phenyl substituted with alkanoylamino having 1 to 3 carbon atoms include 4-acetamidophenyl.

There are two steps to obtain compound (I). One of them is a compound represented by the general formula (U) (in which R2p R3 has the same meaning as above) and a compound represented by the general formula (R3-8-8, /NH2' (@(in the formula, R
3 has the same meaning as above) or its acid addition salt, or 4-amino-1,2-dithiolane hydrochloride under basic conditions in an inert solvent, usually at room temperature from 0°C. This is the way to do it.

As acid addition salts, hydrochlorides, sulfates, nitrates, etc. are used.

As the inert solvent, methanol, ethanol, impropatol, methylene chloride, chloroform, acetonitrile, tetrahydrofuran, dioxane, dimethylformamide, etc. are used alone or in combination. Triethylamine, pyridine, etc. are used as a base for basic conditions. The reaction is carried out using a solvent, the compound used (I
I), (It varies depending on the type of powder, but usually takes several tens of minutes to several hours, and when using 4-amino-1,2-dithiolane, it usually takes several hours to several days. After the reaction is complete, water is injected.) After extraction with chloroform or ethyl acetate, the extract is concentrated under reduced pressure and purified by column chromatography, preparative chromatography, recrystallization, etc.In addition, 4-amino-1,2-dithiolane hydrochloride The synthesis method is chemical
Abstracts 65. 18061 (1965).

In compound (1), the general formula (■ [in the formula, R3' is a straight chain or branched alkyl or cycloalkyl having 1 to 7 carbon atoms (the alkyl or cycloalkyl has 1 to 3 hydroxyl or 1 (optionally substituted with amino), -(OH2)nC!02R4 (, n is 1, 2 or 3, R4 is methyl or ethyl), or phenyl substituted with alkanoylamino having 1 to 6 carbon atoms. and R1 + R2 have the same meanings as above] is a compound represented by the general formula (IV) (wherein R1 and R2 have the same meanings as above) obtained by the above method, and a compound represented by the general formula (IV) (wherein R1 and R2 have the same meanings as above). V) A compound represented by R3'5H(V) (wherein R31 has the same meaning as above) or a base addition salt thereof (e.g., sodium salt, potassium salt) in an inert solvent, usually at room temperature from 0°C. The solvent used in the reaction is an organic solvent such as methanol, ethanol, isopropanol, acetonitrile, dimethylformamide dimethyl sulfoxide, or water, which may be used alone or in combination.

Although the reaction varies depending on the solvent and the type of compound (V) used, it is usually completed in several minutes to several hours. After the reaction is complete, water is injected, extracted with chloroform or ethyl acetate, and the extract is concentrated under reduced pressure, followed by column chromatography.
Purify by preparative thin layer chromatography, recrystallization, etc., or inject chloroform or ethyl acetate, extract with water, concentrate the solution under reduced pressure, and then purify by column chromatography, etc. For methods of disulfide synthesis and utilization using such 2-pyridyl disulfide, see Biochem, J.

173, 723 (1978), Bioc.
hem, Biophy, Ree.

Oomm, 101, 599 (1981)
, Chancer Ree,, 42°457
(j982), Nature, 290.145 (
1981) and others.

Next, Table 1 shows the minimum inhibitory concentration of compound (1) against various bacteria (skin/-1 agar dilution method PH7.0).

The symbols of compounds in Table 1 indicate the following compounds.

7-IN, 7'-N-dithiodiethylenesimatomycin D 7-N, 7'-N-dithiodiethylenesimatomycin E 7-N-(2-aminoethylditheoethyl) mitomycin D 0 7-N-(2 -pyridyldithioethyl) mitomycin O 7-N-(2-pyridyldithioethyl)porphyromycin 7 - Tomycin D 0 7- 0 0 ONE2 7-N-ethyldithioethyl mitomycin 7-N-propyldithioethyl mitomycin C 7 -N-(2-hydroxyethyl, dithioethyl) mitomycin C 7-N-(4-acetamidophenyldithioethyl) mitomycin C 7-N- (4-1.2-dithiolyl)
Mitomycin C7-N-(4-1.2-dithiolyl)
Porphyromycin 7-N- (4-1.2-dithiolyl)
Mitomycin D 7-N- (4-1.2-dithiolyl)
Mitomycin E 7-N-(2-pyridyldithioethyl)mitomycin E ○ 7-N-(2,3-dihydroxyclopyldithioethyl)mitomycin C 7-N-methoxycarbonylmethyldithioethylmitomycin C 7-N-cyclohexyldithioethyl Theoethyl mitomycin C 1st Table MM-0: Mitomycin C OA Candida albicans ATOO10
231SF Streptococcus faecalis A
TOO10541SA Staphylococcus aureus ATC! O/+538PEC Escherichia coli ATOO26BS Bacillus subtilis 10
707Pv Frotheus Vulgaris ATC! 0689
7SS Shigella Zonei ATC! 09290EI
T Salmonella taihosa ATOC9992K
P pLevsiella pneumoniae ATOC10
[031] Next, examples of the present invention will be shown.

Example 1 Preparation of 7-N,7'-N'-dithiodiethylenesimatomycin D: 12.9 mf of cystamine dihydrochloride is dissolved in 3-methanol and 50 μt of triethylamine is added with stirring. Add mitomycin B40ap in several portions and stir at room temperature for 12 hours.

Add water to the reaction solution and extract with chloroform. After drying the extract with sulfuric acid (IJ), the solvent is distilled off under reduced pressure, and the resulting oil is purified by silica gel column chromatography. Chloroform-methanol (4:1;
v/v) as a developing solvent and concentrate the eluted portion to obtain the title compound 42■ as green crystals (yield 93%).
.

Melting point: >220°C decomposition elemental analysis, H5, 43, 051.94, N1 4,
0 2% (as 034H42N8010E2, N5.37%.

C51, 90, N14.249J) IH-NMR (py-c15) δ2.10 (3H, s)
, 2.13 (3H, S), 2.22 (IH, d), 2
．． 47 (IH, d).

2.91 (2H, t), 3.6/1 (IH, a), 58
7 (2H.

q), 4.23 (iH, da), 4.44 (IH, d)
.

5.21 (IH, t), 5.47 (IH, ad'), 7
．． 26 (IH, t), 7.46 (2E, br, a)
Engineering R (KBr) 3420, 3250, 1703, 154
3゜1503.1453.13426n'' Example 2 Preparation of 7-N,7'-N'-dithiodiethylenesimatomycin E: Dissolve 12.4 η of cystamine dihydrochloride in methanol 2- and add 50 μt of triethylamine with stirring. Add mitomycin J40 in several portions and incubate at room temperature.
Stir for 0 hours. Add water to the reaction solution and extract with chloroform. After drying the extract over sodium sulfate, the solvent is distilled off under reduced pressure, and the resulting oil is purified by silica gel column chromatography. The eluted portion was concentrated using chloroform-methanol (95: '5. v/v) as a developing solvent, and the title compound 43 was obtained as a black powder.
'Hi was obtained (yield: 96 H).

Melting point: 132-5 to 134°C Elemental analysis N5.74, 053.21%, N1147
% (C!xeH4sNso+osz and CN5.6
9%.

C5506chi, N13.75%) 'H-NMR (ODO43) δz, o1 (3a, S),
2.22 (IH.

cl), 241 (3H, 8), 2.37 (IH, c
od).

2.85 (2H, t), 3.30 (3H, s), 3.5
7 (1'H.

da), 3.86 (2H, q), 4.02 (IH,, d
).

4.45 (IH, cud), 4.74 (2H, br,
s), 4.80 (IH, aa), &47 (IH, t)
.

Engineering R (KBr) 3300,2930. 1716
,1553°1510,' 1445. '1555
．． 1050c+++-'Example 3 Preparation of 7-N-('2-aminoethyldithioethyl)mitomycin D: Cystamine dihydrochloride 10311! f methanol 2.5
Triethylamine 0.4-
drip. Add 40 kg of mitomycin B in several portions and stir at room temperature for 40 minutes.

Pour water and extract with chloroform. After drying the extract with anhydrous sulfuric acid, the solvent was distilled off under reduced pressure, and the resulting oil was subjected to column chromatography using Diaion Hp-20 (trade name of adsorption resin, manufactured by Mitsubishi Kasei). refine.

The eluted portion with water-methanol (1:4:v/v) was concentrated to give the title compound 2 as a dark green paste.
3■ is obtained (yield 43%).

Elemental analysis N5.82.04 & 67, N14.70 (0
19H27N505S2 N5.80.
C4a6o.

N 1 4.9 1 ) ' H-NMR(py-(15) δ2.oa(xa
, θ), 2.13 (3H.

s), 2.23 (IH, da), 147 (IH, a).

2.78-3.17 (6H, nt), 5.66CIH,
(1).

586 (2J q'), 4.21 (IH,'dd)
, '4.43 (IH, cl), 5.19 (IH, act
), 5.46 (IH, act).

7.23(1'H,t)t 7.45(2H,br,
s) Engineering R (KBr) 3280,1707. 1/
)29. 1547°1511.1451,1332,
1071 cm2L Example 4 Production of 7-N-(2-pyridyldithioethyl) mitomycin C: 50 μm of pyridyldithioethylamine-2 hydrochloride described in JP-A-55-136261 was added to 2.5 μm of methanol.
0.2-triethylamine dissolved in d was added dropwise with stirring. Mitomycin A40■ was added in several portions and stirred at room temperature for 5 hours. Water is poured into the reaction solution and extracted with chloroform. After drying the extract over sodium sulfate, the solvent is distilled off under reduced pressure, and the resulting oil is purified by silica gel column chromatography. The eluted portion was concentrated using ethyl acetate-methanol (9s:s; v/v) as a developing solvent to obtain the title compound 54 as a black-purple powder.
η is obtained (yield 94%).

Elemental analysis H5,06,052,58,N 13.6
3 (022N25 N5 o582 as H5,
OCI, 0 52.47°N 1591) IH-NMR (py4g) δ2.06 (3H, 8),
2.7iS (IH, br ), 1 mouth 8 (2H, t),!
c14 (IH, br).

3.6o (1H, aa), 3.9o (1a, q),
4o.

(IH, da), 4.53 (IH, d), 5.06 (1
H.

t), 5.40 (IH, da), 7.06 (2H, m)
.

z61 (2H, m), a66 (1u, m) engineering R (KBr
) 3290, 2930, 1713, 1632°15
53.1505, 1444, 1414, 1330°10
61,7556n-" Example 5 Production of 7-N-(2-pyridyldithioethyl)porphyromycin: Pyridyldithioethylamine dihydrochloride 50 and mitomycin P40q were prepared as black-purple powder in the same manner as in Example 4. The title compound 56 is obtained (yield: 98 cm).

Elemental analysis H5,50,05L41. N 13.
32 (023HzyNsOsSz as H5,26
,053,37°N I&53) IH-NMR (a: oat3) δ1.94 (3H, s)
, 2.27 (3H, s), 2.27 (2H, m), 2.
'? 6 (2H, t).

A18 (3H, s), 545 (IH, dd), 3.57
(IH, da), ! L86 (2H, q), 4.24 (
IH, d).

4.35 (IH, aa), 4.72 (IH, aa), 4
．． 73 (2H, br, s), 7.10 (IH, br,
), 715CIH.

m), 7.50-7.72 (2J m), δ59 (1H
, m) Engineering R (KBr) 3290, 2930, 171
9.1632°1557.1508,1445,132
6,1060°758tM-1 Example 6 Production of 7-N-(2-pyridyldithioethyl) mitomycin D: 50 μ of pyridyl dithioethylamine dihydrochloride and 40 μ of mitomycin B were prepared in the same manner as in Example 4. The title compound 57Ilv is obtained as a colored powder (yield: 99cm).

Elemental analysis H5,03,052,51,N IA69
(022H2S ~5 o5B2 as Hs, o
o, 0 52.47°N 1591) ”H-NMR (CDO43) δ1,91 (3H,8),
2.27C5H,s), 2.27(2H,m), 2.9
6 (2H, t).

3+49 (IH, br, d), ! 1.71 (1H, t
), 3.84 (2H, q), 4.14 (IH, a), 4
．． 48 (ILbr, ), 4.72 (2H, a), 4
81 (2H,br,).

7.02 (IH, br,), 7.10 (IH, m),
7.50-7.70 (2H, m), δ56 (IJ m
) Engineering R (KBr) 3270, 1713, 1631,
1601°1548.1510,1450,1414,
1552゜1113.1058,758cyn-" Example 7 Production of 7-N-ethyldithioethyl mitomycin C: 7-N-(2-pyridyldithioethyl) mitomycin 050++y synthesized in Example 4 was dissolved in methanol 2.5
- dissolve in and add 8 μt of ethanethiol. After stirring at room temperature for 10 minutes, saturated aqueous sodium bicarbonate was poured into the mixture, followed by extraction with chloroform. After drying the extract over sodium sulfate, the solvent is distilled off under reduced pressure and the resulting oil is purified by silica gel column chromatography. Concentrate the eluate using chloroform-methanol (96:4゜v/v) as a developing solvent, and drop the chloroform solution of the residue into cyclohexane to obtain the title compound 43fiv as a grayish-blue powder (yield: 96cm). .

Elemental analysis H5,7905[129N 12.16(
019Hzs ~405 Bz as H5,'7
6 C50,2ON tz3s) 'H-NMR (CDO23) δ1.33 (5H, t),
2.03 (3H, s), 2.67 (2H, q), 2.8
4 (2H, t).

2.85 (2H, m), A21 (5H, 8), 151 (
IH.

cud), A62 (IH, da),! L87 (2H, q
); 428 (IH, a), tso (1H, aa), 4.
68 (2H, br, s), 4.71 (IH, dd), -
6,49 (IH.

br, t) Engineering R' (KBr) 3270.1718,1633
．． 1'554゜1507.1447,1328,106
Example 8 Preparation of 7-N-propyldithioethyl mitomycin C 050 my of N-(2-pyridyldithioethyl) mitomycin and 9 μt of propanethiol were prepared in the same manner as in Example 7 to produce the title compound as a gray-blue powder. 29 capes are obtained (yield 62 cm).

Elemental analysis H6,0,3C! , 51.32 M
11.75 (as 020H28N40SS2 H'
6.02 0 51.26N11.96) IH-NMR (aDcz,) α99 (5J t),
1.71 (2H.

5extet), 2.03(3H,s), 2.6
8 (2H, t).

2.83 (2H, t), 2.85 (2H, m), 3.
21 (5H.

s), ! L51 (IH, dd), 560 (IH, aa
).

584 (2H, q), 428 (IH, d), 4.50 (
IH.

ad), 4.6B (2H, br, s), 4.71' (I
H, dd).

6.48 (IH, br, t) Engineering R (KBr) 3290,1721. 1634,
1558°1509.144B, 1327. 1060
cfn-1 Example 9 Production of 7-N-(2-hydroxyethyldithioethyl) mitomycin C Using near-N-(2-pyridyldithioethyl) mitomycin 050M1i and 7 μt of 2-mercaptoethanol, the same procedure as in Example 7 was carried out. The title compound 4111v is obtained as a gray-blue powder (yield: 88 cm).

Elemental analysis H5,6004&12 N 11.62
(0111HHIN4 o6s, as H5,5
70,4a5ON11,91) IH-NMR (cDcts) 2.62 (3H, s)
, 2.80-2.97 (2H, m), 2.86 (2H
, t), 2.90 (2H, t).

121 (3H, 8),! L51 (IH, ad), 5.5
9 (IH, ad), ! L87 (2J q), Engineering 88 (
2Jt).

4.27 (IH, cl), 4.50 (IH, da), 4
．． 70 (1H, ad), 4.78 (2J br, s)
, 6.49 (IH.

br, t) Engineering R (KBr) 3430.5280,1712.
1652°1551.1510,1448,1328
, 1060cm ”Example 10 Production of 7-N-(4-acetamidophenyldithioethyl) mitomycin C -N,-(2-pyridyldithioethyl) mitomycin C30Wf and 4-acetamidothiophenol 17w
In the same manner as in Example 7, the title compound 38 was obtained as a gray-blue powder (yield: 69).

Elemental analysis H5, 2505569 N 12.30
(C!zs Hzs Ns Og St as H5
,22,05165°N 12.51)'H-NMR(py-ds) 2.06 (3H,s
), 2.16 (,3Js), 2.75(IH,b
r, d), XQ3(2H, t).

WLl5(IH, (1), 523 (3H, s), Δ59
(IH.

br, a), x9o (2n, q), x99 (1n, aa
).

4.52 (IH, (1), 5.04 (IH, aa), 5
．． 39 (1H, ad), 7.13 (IH, br, t
), 7.66 (2H.

br. s), Z59-a03 (4H, AA'BB'
), 1a76 (IB, br, s) Engineering R (KBr) 3290. 1713, 1655
,1589°1553.1510,1323,1062
．． 755cyy+"Example 11 Production of 7-N-(4-1,2-dithiolyl)mitomycin C: 14 ml of 4-amino-1,2-dithiolane hydrochloride was mixed with dimethylformamide-methanol (4:1°v/v) 5
- in addition to triethylamine 0. Add IWLl and stir at room temperature. Add mitomycin 83019,
Stir at room temperature for 3 days. After injecting water and extracting with chloroform, the solvent is dried over sodium sulfate and distilled off under reduced pressure. The remaining oil was diluted with ethyl acetate-acetone (7:3
．． (v/v) was purified by silica gel thin-layer chromatography using a developing solution to obtain the title compound 32sy as a dark brown paste (yield: 85cm).

Elemental analysis H&09 C49,42N 12-51
(H5,070 as 01BH21”40581
49,5ON 12.78) MS (l knee MS) m/z 438 (Mzu, 406
,395°577 275 IH-NMR (aDaz3) δ z, ol(+a,
s), 2.82 (1, H, aa), 2.91 (IH,
a), 3.11~x+a (4H.

m), 21 (3H, 8), &51 (1H, dd).

! L61 (IH, d4), 4.24 (IH, d), 4.
50 (IH, dd), 4.72 (IL ad), 4.
73 (2H, da).

5.24 (IH, m), 6.38 (1H, br, d) Engineering R (KBr) 3280. 2930. 171
8. 1/+33°1561, 1501. 1447
．． 1338. 1059°758 cm-' Example 12 Production of y-N-(4-1,2-dithiolyl)porphyromycin: Performed using 4-amino-1,2-dithiolane hydrochloride 16 and porphyromycin 30η Similarly to example 11,
The title compound 28'q is obtained as a dark brown powder (yield: 7
5%).

Elemental analysis H5,37,050,49,N 12.2
[1 (019H24A405 S2 as H5j
5. 0 50.43°N 12.5B) MS (FD-MS) m/z 453 (M+H)+
'H-NMR (cDaz3) δ 2.00 (3H,
s), 2! :27 (3L s), 2.27 (2H, m
), 505-435 (4H.

m), hls (sH, s), 3.45 (IH, aa).

xs6 (1a, aa), 420 (IH, a), 4.34
(IH, da), 4.70 (IH, da), 4.71 (
2H, br.

8), 5.24 (IH, m), 6.39 (IH, b
r, a) Engineering R (KBr) 3450, 3350,
3250, 2960°1722, 1699. 163
9. 1566, 1508°1341.1055.7
44cm-1 Example 13 Production of 7-N-(4-1,'2-dithiolyl)mitomycin D: Produced in the same manner as in Example 11 using 4-amino-1,2-dithiolane hydrochloride 14η and mitomycin B50Tq. The title compound 32η is obtained as a brown powder (85% yield).

Elemental analysis H5,08,049,36,N 12.5
9 (as C'18 A22 B'4 o5S2
H5,07,049,30°N 12.78) MS (FD-Ms) m/z 439 (M+H)+IH
-NMR (CDC73) δ 1.99 (3H, 8)
, 2.27 (3H, 8), 3.05-440 (4H, m
), 150 (IH.

aa), x7o(1a; t), 4;12(IH,a
).

4°49 (IH, br, s), 4.71 (2H, d),
4.77 (2H, br, s), 5.24 (IH, m),
A39 (IH.

br, (1) Engineering R (KBr) j350,2960,1:A3,1
6/+0°1631.155! l, 1501, 1342
, 1052crn-1 Example 14 Production of 7-N-(4-1,2-dithiolyl)mitomycin E: From 4-amino-1,2-dithiolane hydrochloride 13η and mitomycin J30■, in the same manner as in Example 11. The title compound 27 is obtained as a dark brown powder (yield 72%).

Elemental analysis H5,36,C! 50.44. N
12.09 (019A24 A40S S2 as E 5.35. C5CL43°N 12
．． 38) IH-NMR (CDC13) δ 1.99 (3H,
θ), 2.22 (IH, a), 132 (3H, 8), 2
．． 38 (IH, aa).

504-3.39 (4H, m), 3.31 (3B, =)
.

558 (IH, ad), X81 (IL clcl),
3.99 (IH, a), 4.45 (IH, da), 46
7 (2H.

br,,G), 4.79 (IH, dd), 5.24 (I
H, m).

6.34 (IH, br, d) Engineering R (KBr) 3560.295G, 1717,
1631°1557.1501. j444,133
3,1048゜750cn' Example 15 Production of 7-N-(2-pyridyldithioethyl) mitomycin E: Pyridyldithioethylamine dihydrochloride 50 to tomitomycin J41] The title compound 55 is obtained as a powder (yield 96%).

Elemental analysis A5.28. 0 52.47. , N
1i26 (023H27N505S2 as H
5, 26, C! 53.37°N 13.53) "H-NMR (CDC/6) δ 1.93 (3H,
s), 2.21 (IH, cl), 2.32 (3H, e)
, 2.37 (LH, dd).

2.96 (2H, t), xsl (sa, 8), A37 (
IH, ad), 181 (IH, ad), 3.85 (2H
,q).

ζ02 (IH, a), 44s (1H, aa), 4.
/16 (2H, br, s), 4.82 (IH, da),
A99 (IH.

br, t), 7.11 (I H, m), 7.
58 (2H, nt).

3s7 (1u,'m) 1 piece (KBr) 3280. 2950. 17
17. 1631°1551.1507,1445,1
413,1332゜1112.1047.757σ "1 Example 16 Production of 7-N-(z, 3-dihydroxypropyldithioethyl) mitomycin C Near-N-(2-pyridyldithioethyl) mitomycin 040■ and 1-mercapto -2,6-Propanediol Z1μt A black paste was obtained in the same manner as in Example 7, and further freeze-dried to obtain the title compound 3 as a black powder.
3■ was obtained (yield: 83cm).

Elemental analysis H5,62,047,96,N 11.0
1 (026H2B N4 o7s2 as H5,
64, C4799°N11.19) 'H-NMR (py-4g) δz1x (xH, 11),
2.73 (IH, m), five o! +(2H,t), 3
．． 12 (IH,,a).

5.22 (5B, g), 557 (2H, d),! L59
(IH.

aa), xa3-4.11 (3H, m), 4.08 (2
H, a).

4.44 (IL m), 4.52 (IH, d), 5.0
3 (IH.

m), 5.38(IH, (1(1), 7.28(i
H2br,).

7.57 (2H, br,) Engineering R (KBr)' 3430.52B0,2930,
1901°1652.1553,1510,1449.
1332°1064, 755. crn-1 Example 17 Production of 7-N-methoxycarbonylmethyldithioethylmicin C: 05 (] mg of 2N-(2-pyridyldithioethyl) mitomycin and 8 μL of methyl thioglycolate a were prepared in the same manner as in Example 7. The title compound 3 was obtained as a gray-blue powder.
4 pieces were obtained (yield: 69 pieces).

Elemental analysis H5, 28, C4a21. N 10.
97 (OzoH2gL 07 B2 as H5,
26,04a18゜B111.24) IH-NMR (cDaz3) δ2.02 (3,H,S
), 2.84 (2H.

br, e), 2.95 (2H, t), 3.21 (3H,
s).

3.51 (2H, s), 3.52 (IH, 'br, d)
, 560 (IH, ad), ni 76 (3H, e), 58
9 (2H, q).

4.28 (IH, a), 450 (IH, ad), 4.7
3 (IH, ad), 4.74 (2H, br, s), &4
7 (1H.

br, t) Engineering R (KBr) 3280, 2930, 1723, 1
554°1508, 1445. l 330.106O
CA-' Example 18 Preparation of 7'-N-cyclohexyldithioethyl mitomycin C 050 q of -N-('2-pyridyldithioethyl) mitomycin was dissolved in 2.5 d of methanol, 12.1 μt of cyclohexanethiol was added, After stirring at room temperature under a nitrogen stream for 12 hours, the mixture was treated in the same manner as in Example 7 to obtain 49.5 W of the title compound as a gray-blue powder (yield: 98%).

Elemental analysis H6,55,054,33,N 1O-7
6 (as 023H3□N4o5s2 H6,34
, Q 54.31°N11.01 1H-NMR (ODO63) δ1.02~2.20 (
10H, m).

2.03 (3H, l1lI), 2.68 (IH, br
, ), 2=82 (4H, m), 3.20 (3H, 8)
, 3h51 (IH, da).

161 (IJ aa),! +, 86 (2H, q), 4.
28 (IH, d), 4.50 (IH, dd), 4.72
(IH, d4).

4.83 (2H, br, s), 6.49 (IH, br
,t) Engineering R (KBr) 3saa, 295o, 2
850,1718°1633.1558,1509,1
448.1329゜1062cIn-' Patent Applicant (102) Kyowa Rakko Kogyo Co., Ltd. Procedural Amendment May 13, 1980 Dear Commissioner of the Japan Patent Office 1. Indication of the case 1988 Patent Application No. 50186 2. Name of the invention Mitomycin derivatives and their manufacturing method 3, relationship with the amended case Patent applicant postal code 100 Address 6-1 Otemachi-chome, Chiyoda-ku, Tokyo Name
(102) Contents of amendment in Pack 5 of Detailed Description of the Invention in Kyowa Hakko Kogyo Co., Ltd. Specification (1) "Azole" in line 5 of page 8 is corrected to "azolyl".

(In the figure on page 2116g, r Ns −s...” is replaced with “＼
/S-8...''.

f31 In the diagram on page 16, "s-s..." is replaced with "small,, t.
S 8..." is corrected.

(4) On page 17, A! , page 18 m, n compound group r4
-t,2-dithiolyl” to “1,2-dithiolane-4-
Corrected to "il".

(Page 5133, line 7, page 34, line 15, page 35, line 15.

r4-x,z-dithiolyl” on page 36, line 14 is replaced with “1,2
-dithioran-4-yl".

+6) Correct rz, 62J on page 31, line 16 to rz, o2j.

(7) Correct "1901" on page 39, line 13 to r1701J.

Mr. Commissioner of the Japan Patent Office 1. Indication of the case Patent Application No. 50186 of 1982 2. Name of the invention Mitomycin derivative and its manufacturing method 3. Person making the amendment Relationship to the case Patent applicant Postal code 100 Address Ote, Chiyoda-ku, Tokyo Town-chome 6-inspection 1 name (
102) Kyowa Hakko Kogyo Co., Ltd. (Location: 03-201-7
211 extension 2751) Contents of amendment in Column 5 of Detailed Description of the Invention in the Specification (1) Page 19 After the description of compound r, the following description is added.

[7-N-(2-aminoethyldithioethyl)porphyromycin” (2+20 pages rrJ and “
Add the following values for "S" and 6 during the description of MM-cl.

(The following example is added after the description of Example 18 on page 3141.

Example 19 Preparation of 7-N-(2-aminoethyldithioethyl)porphyromycin: Dissolve 100 ml of cystamine dihydrochloride in 2.54 ml of methanol, add 0.4 d of triethylamine, and stir.Mitomycin F 40149 was added in several portions and stirred at room temperature for 15 minutes, then the drained water was injected and extracted with chloroform.The extract was dried over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and the resulting oil was subjected to silica gel chromatography. Chloroform-methanol (3:
2; V/V) was concentrated to obtain the title compound 12117 as a black base (yield 23%).

Elemental analysis H6,07, C49,76, N 14.23
(T(6,04,C4 as C20H29NSO5S2
9,671N 14.48)H-NMR (1:17-
as) δ2.14 (3H,8), 2.16 (LH
, m).

2.24 (3H, 8), 2.53 (IH, d), 2
．． 80-3.19 (6H.

m), 3.19 (3H, s), 3.52 (IH, a
d), 3.94 (IH, aa), 3.94 (2H1
ff), 4.45 (IH, d).

4.77 (IH, (ld), 5.31 (IH, d (1
), 7.27 (IH.

br, t), 7.64 (2H, br, e)IR(K
Br) 3360.1714.1633.1554.
1510°1459, 1328.1062 cm-”
J Procedural Amendments Mr. Commissioner of the Japan Patent Office 1. Indication of the case Patent Application No. 50186 of 1982 2. Name of the invention Mitomycin derivatives and their production method 1. Relationship with the case Patent applicant Postal code 100 Address Tokyo Chiyoda-ku Otemachi-chome 6-1 Name
(102) Column 5 of the detailed description of the invention in the specification of Kyowa Hakko Kogyo Co., Ltd., contents of the amendment All amendments made in the procedural amendment dated May 31, 1988 will be deleted.

Procedural Amendment Written July J, 1980 Mr. Commissioner of the Japan Patent Office 1. Indication of the case 1988 Patent Application No. 50186 2. Name of the invention Mitomycin derivatives and their manufacturing method 3. Person making the amendment Relationship with the case Patent applicant Postal code 100 Address 6-1 Otemachi, Chiyoda-ku, Tokyo Name
(102) Kyowa Hakko Kogyo Co., Ltd. (location: 03-201
-7211 extension 2751) Add the following sentence between column 5 of the detailed description of the invention in the specification, content of amendment, page 10, and between the bottom line and line 8.

[In this case, a compound (■) in which R3 is a linear or branched alkyl or cycloalkyl having 2 to 7 carbon atoms substituted with one amine group (hereinafter referred to as compound (■')) When more than twice the mole of compound (I) is used, the compound (I
A product obtained by reacting I) and compound (■') in a ratio of 1=1 was obtained as the main product (see Example 3), and approximately 0.5 times the molar amount of compound (■') relative to compound (n) was obtained. When used, a product obtained by reacting compound (If) and compound (■') in a ratio of 2:1 is obtained as the main product (see Example 1). "hand
Continuing Amendment Written July 9, 1972 Director-General of the Patent Office 1. Description of the case 1982 Patent Application No. 50186 2. Name of the invention Mitomaitsuno derivative and its manufacturing method 3. Case of the person making the amendment Relationship with Patent Applicant Postal Code 100 Address 6-1 Otemachi-chome, Chiyoda-ku, Tokyo Name (102) l@Wahokko Kogyo Co., Ltd. Contents of the Detailed Description of the Invention Column 5 of the Specification (1) Correct "Porphyromynon" in the bottom two lines of page 34 to "Mitomynon F".

(2) r C52, 47J in line T1 on page 37 to 1'C53
, 47j.

Procedural amendments dated February ζ, 1980, Commissioner of the Japan Patent Office 1, Indication of the case, Patent Application No. 50186 of 1982, 2, Name of the invention, mitomaine derivatives and its manufacturing method 3, Person making the amendment Relationship with the case Patent application Postal code: 100 Address: 6-1 Otemachi-chome, Chiyoda-ku, Tokyo Name (
102) Kyowa Hakko Kogyo Co., Ltd. (Location: 03-201-7
211 Extension 2751 > Detailed explanation of the invention in the specification ’-゛’5, Contents of amendment (3) Page 36, line 6, after r)l, s), and j, r2.2
7 (2H, m), join J.

(4) Page 39, line 8, r3.37 (2H, d) J to r3
．． 35 (2H,,i+), corrected to J.

Claims (1)

[Claims] (1) General formula [wherein R3 is a straight chain or branched alkyl or cycloalkyl having 1 to 7 carbon atoms (the alkyl or cycloalkyl may be substituted with 1 to 5 hydroxyl or 1 amino good), -(CH2)nCO2R4 (n is 1,2
or 3, R4 is methyl or ethyl), pyridyl,
Phenyl substituted with alkanoylamino having 1 to 2 carbon atoms, or 1R1+ R
2 are the same or different and are hydrogen or methyl, but are not hydrogen at the same time. W represents an α or β bond. Regardless of the above definition, when carbamyloxymethyl at position 9 is α, R3 has 1 to 7 carbon atoms substituted with an amino group.
When the straight-chain or branched alkyl or carbamyloxymethyl at the 9-position is β and R1 is methyl, R2 is not hydrogen. ) A mitomycin derivative represented by 12) General formula U (In the formula, R1, R, are the same or different and are hydrogen or methyl, but are not hydrogen at the same time. Regardless of the above definition, carbamyloxymethyl at the 9-position is β and η\ When R1 is methyl, R2 is not hydrogen.) and R's'-f3E [wherein R3' is a straight chain or branched alkyl or cycloalkyl having 1 to 7 carbon atoms (the alkyl or Cycloalkyl is optionally substituted with 1 to 3 hydroxyl or 1 amino-(aH,)nco2R4 (n is 1,
2 or 3, R4 is methyl or ethyl), or phenyl substituted with alkanoylamino having 1 to 3 carbon atoms. 2 t R1' has the same meaning as above) A method for producing a mitomycin derivative.