New mitomycins are produced by synthetic processes. The compounds exhibit broad spectrum antibacterial activity and are useful as antibacterial agents.

BACKGROUND OF THE INVENTION 
1. Field of the Invention 
This invention relates to new mitomycins and processes for production 
thereof. 
2. Description of Prior Art 
Mitomycins are generally known as compounds having anti-tumor, 
antibacterial activity. Representative mitomycins include mitomycin A, 
mitomycin B, mitomycin C, and porfiromycin, which are described in the 
Merck Index (Ninth Edition). In addition, Japanese Published Unexamined 
Patent Application No. 122797/1979 describes other mitomycins viz. 
mitomycin D and mitomycin E. These known mitomycins may be obtained by 
culturing a strain of Streptomyces caespitosus and have the following 
chemical structures: 
______________________________________ 
##STR1## 
X.sub.A 
9 10 R.sub.D R.sub.E 
______________________________________ 
Mitomycin A OCH.sub.3 CH.sub.3 
H 
Mitomycin B OCH.sub.3 H CH.sub.3 
Mitomycin C NH.sub.2 CH.sub.3 
H 
Mitomycin D NH.sub.2 H CH.sub.3 
Mitomycin E NH.sub.2 CH.sub.3 
CH.sub.3 
Porfiromycin 
NH.sub.2 CH.sub.3 
CH.sub.3 
______________________________________ 
Various derivatives of these compounds are also known. For example, 
1a-N-acetyl-mitomycin C (U.S. Pat. No. 3,514,452), 1a-N-butyryl-mitomycin 
C (U.S. Pat. No. 3,514,452), 7-ethylamino-7-demethoxy-mitomycin A (U.S. 
Pat. No. 3,514,452), 10-decarbamoyl-mitomycins A, B and C (U.S. Pat. No. 
3,738,998) 1a-N-acetyl-10-decarbamoyl-mitomycin C (Japanese Published 
Examined Patent Application No. 17,279/1974), 
1a-N-acetyl-10-decarbamoyl-10-p-toluene-sulfonyl-mitomycin C (Japanese 
Published Examined Patent Application No. 37,639/1972), 
10-decarbamoyl-10-p-toluene-sulfonyl-porfiromycin (Japanese Published 
Examined Patent Application No. 37,639/1972), etc. are known. In addition, 
mitomycins having a double bond between 9- and 10-positions, other than 
those described in the present specification, are disclosed in commonly 
owned U.S. Patent Application Ser. No. 58,670 filed on July 18, 1979. 
While the known mitomycins exhibit good activity, new antibacterial 
compounds are always in demand. To this end, the present inventors have 
found new mitomycins which exhibit good antibacterial activity. 
SUMMARY OF THE INVENTION 
In accordance with the present invention, new mitomycins are produced 
having the general formula [I] (hereinafter sometimes referred to as 
Compound [I]. Compounds of other formulae are similarly identified.) 
##STR2## 
wherein X is an amino group, a substituted amino group, a hydroxy group or 
an alkoxy group; Y is a hydrogen atom, a methyl group, --COR.sup.1 wherein 
R.sup.1 is a hydrogen atom, an alkyl group, a substituted alkyl group, an 
aryl group or a substituted aryl group, or --CH.sub.2 CH.sub.2 Z wherein Z 
is an alkoxycarbonyl group, an acyl group or a cyano group; and R.sub.A is 
a hydrogen atom and R.sub.B is --CH.sub.2 OH or --CH.sub.2 OSO.sub.2 
R.sup.2 wherein R.sup.2 is an alkyl group, a substituted alkyl group or 
an aryl group, or R.sub.A and R.sub.B combine to form =CH.sub.2. 
Notwithstanding the above definition, when R.sub.A is a hydrogen atom and 
R.sub.B is --CH.sub.2 OH, Y is --CH.sub.2 CH.sub.2 Z, when R.sub.A is a 
hydrogen atom and R.sub.B is --CH.sub.2 OSO.sub.2 R.sup.2, Y is a hydrogen 
atom or --CH.sub.2 CH.sub.2 Z and when Y is a methyl group, X is a hydroxy 
group and R.sub.A and R.sub.B combine to form =CH.sub.2. 
The invention also pertains to various synthetic processes for producing 
Compound [I]. 
The compounds of the present invention (Compound [I]) have broad 
antibacterial activity and are, therefore, useful to clean and sterilize 
laboratory glassware and surgical instruments. The compounds may also be 
used in combination with soaps, detergents and wash solutions for sanitary 
purposes. Compound [I] may also be useful as medicaments or intermediates 
in the preparation of other mitomycin derivatives having similar activity. 
DESCRIPTION OF THE INVENTION 
Compounds of the present invention are represented by the general formula 
[I]: 
##STR3## 
In formula [I], X is an amino group, a substituted amino group, a hydroxy 
group or an alkoxy group. The term "alkoxy" group includes an alkoxy group 
having 1-4 carbon atoms, e.g., methoxy, ethoxy, i-propoxy, n-butoxy, 
t-butoxy and the like. Y is a hydrogen atom, a methyl group, --COR.sup.1 
or --CH.sub.2 CH.sub.2 Z. In the representation --COR.sup.1, R.sup.1 is a 
hydrogen atom, an alkyl group, a substituted alkyl group, an aryl group or 
a substituted aryl group. The term "alkyl" group includes an alkyl group 
having 1-3 carbon atoms, e.g., methyl, ethyl, i-propyl and the like; the 
term "substituted alkyl" group includes chloromethyl and the like; the 
term "aryl" group includes phenyl and the like; and the term "substituted 
aryl" group includes p-methoxyphenyl and the like. In the representation 
--CH.sub.2 CH.sub.2 Z, Z is an alkoxycarbonyl group, an acyl group or a 
cyano group. The alkyl group of the term "alkoxycarbonyl" group includes 
an alkyl group having 1-4 carbon atoms, e.g., methyl, ethyl, t-butyl and 
the like; and the term "acyl" group includes --COR.sup.4 wherein R.sup.4 
is hydrogen or an alkyl group having 1-3 carbon atoms, e.g., methyl, 
ethyl, i-propyl and the like. In formula [I], R.sub.A is a hydrogen atom 
and R.sub.B is --CH.sub.2 OH or --CH.sub.2 OSO.sub.2 R.sup.2 wherein 
R.sup.2 is an alkyl group, a substituted alkyl group or an aryl group, or 
R.sub.A and R.sub.B combine to form =CH.sub.2. In the representation 
--CH.sub.2 OSO.sub.2 R.sup.2, R.sup.2 includes a methyl group, a 
trifluoromethyl group, a p-methylphenyl group and the like. 
Notwithstanding the above definition, when R.sub.A is a hydrogen atom and 
R.sub.B is --CH.sub.2 OH, Y is --CH.sub.2 CH.sub.2 Z; when R.sub.A is 
hydrogen and R.sub.B is --CH.sub.2 OSO.sub.2 R.sup.2, Y is a hydrogen atom 
or --CH.sub.2 CH.sub.2 Z; and when Y is a methyl group, X is a hydroxy 
group and R.sub.A and R.sub.B combine to form =CH.sub.2. 
The most preferred compounds of the present invention (Compound [I]) have a 
double bond between the 9- and 10-positions and are represented by the 
general formula [I'] 
##STR4## 
wherein when Ya is a hydrogen atom, X has the same meaning as defined 
before and when Ya is a methyl group, X is a hydroxy group. 
Specific examples of Compound [I] are set forth in the following Table. The 
physical properties of the compounds as well as processes for synthesis 
thereof are set forth in the examples identified. The compound numbers are 
also sometimes used in the description which follows to identify that 
particular compound. 
__________________________________________________________________________ 
No. of 
com- 
Name of Structural No. of 
pound 
compound formula example 
__________________________________________________________________________ 
1 1a-Nacetyl-10- decarbamoyloxy-9- dehydro- mitomycin 
##STR5## 1 
2 10-decarbamoyloxy- 9-dehydro- mitomycin C 
##STR6## 2,6, 8,11 
3 7-deamino-10- decarbamoyloxy- 9-dehydro-7-hydroxy- mitomycin 
##STR7## 9 
4 10-decarba- moyloxy-9- dehydro- mitomycin A 
##STR8## 10 
5 10-decarbamoyl- 10-methane- sulfonyl- mitomycin C 
##STR9## 7 
6 10-decarbamoyl- 1a-N(2-formyl- ethyl)-10- methanesulfonyl- mitomycin 
C 
##STR10## 4 
7 10-decarbamoyl- 1a-N(2-formyl- ethyl)- mitomycin C 
##STR11## 3 
8 10-decarba- moyloxy-9- dehydro-1a- N(2-formyl- ethyl)- mitomycin 
##STR12## 5 
9 10-decarba- moyloxy-9- dehydro-7- deamino-7- hydroxy- porfiromycin 
##STR13## 12 
__________________________________________________________________________ 
The minimum inhibitory concentration (.mu.g/ml) (agar dilution method, pH 
7.0) of the foregoing compounds against various bacteria identified below 
is illustrated in the following Tables. 
A: Serratia marcescens ATCC 4003 
B: Pseudomonas cepacia ATCC 25608 
C: Staphylococcus aureus ATCC 6538P 
D: Escherichia coli ATCC 26 
E: Bacillus subtilis No. 10707 
F: Proteus vulgalis ATCC 6897 
G: Shigella sonnei ATCC 9290 
H: Salmonella typhosa ATCC 9992 
I: Klebsiella pneumoniae ATCC 10031 
TABLE 1 
__________________________________________________________________________ 
Test Bacteria 
compound 
A B C D E F G H I 
__________________________________________________________________________ 
Compound 
&gt;200 &gt;200 
200 -- 
6.3 200 
-- &gt;200 
50 
Mitomycin 
2.5 &gt;10 
0.16 
5.0 
0.039 
0.078 
1.3 
5.0 0.020 
C 
__________________________________________________________________________ 
TABLE 2 
__________________________________________________________________________ 
Test Bacteria 
compound 
A B C D E F G H I 
__________________________________________________________________________ 
Compound 
&gt;11 
&gt;11 
2.6 -- 0.041 
1.3 -- &gt;11 
0.66 
Compound 
&gt;8.6 
&gt;8.6 
4.3 -- 0.54 
4.3 -- &gt;8.6 
4.3 
3 
Compound 
&gt;150 
&gt;150 
9.4 &gt;150 
0.59 
9.4 -- &gt;150 
4.7 
4 
Compound 
&gt;170 
83 42 -- 42 170 -- &gt;170 
42 
5 
Compound 
&gt;200 
50 50 -- 50 100 &gt;200 
&gt;200 
&gt;200 
6 
Compound 
220 54 3.4 220 
6.8 6.8 110 
54 6.8 
7 
Mitomycin 
5.6 5.6 0.087 
&gt;11 
0.044 
0.087 
5.6 5.6 0.011 
C 
__________________________________________________________________________ 
TABLE 3 
______________________________________ 
Test 
com- Bacteria 
pound A B C D E F G H I 
______________________________________ 
Com- &gt;50 50 13 -- 0.39 25 -- &gt;50 13 
pound 
Mito- 2.5 10 0.039 
&gt;10 0.020 
0.039 
5 2.5 0.0098 
mycin 
C 
______________________________________ 
TABLE 4 
______________________________________ 
Test 
com- Bacteria 
pound A B C D E F G H I 
______________________________________ 
Com- &gt;200 -- 100 200 &gt;0.098 
25 200 50 6.3 
pound 
Com- &gt;200 -- 100 -- 0.195 100 &gt;200 -- 3.1 
pound 
A* 
______________________________________ 
Processes for the production of Compounds [I] are exemplified by the 
following flow diagram. 
##STR14## 
In the above processes, Xa is an amino group, a substituted amino group or 
an alkoxy group (Xa=X where X.noteq.hydroxy). Xaa is amino or substituted 
amino (Xaa=Xa where Xa.noteq.alkoxy). Xab is alkoxy (Xab=Xa where 
Xa.noteq.amino or substituted amino). R.sup.1, R.sup.2 and Z have the same 
meaning as defined before. 
Compounds [I-1]-[I-8] are included in Compound [I]. Compounds [I-2-1] and 
[I-2-2] are included in Compound [I-2]. Compounds [II]-[V] are known 
compounds. Compound [VI] is disclosed in U.S. Patent Application Ser. No. 
58,670, filed July 18, 1979. 
Each of the above processes is described below. 
Process of Compound [V].fwdarw.Compound [IV] 
Included in Compound [V] are Mitomycin C wherein Xa is an amino group and 
Mitomycin A wherein Xa is a methoxy group. A compound included in Compound 
[V] wherein Xa is an alkoxy group and a process for production thereof are 
disclosed in U.S. Pat. No. 3,558,651. A compound included in Compound [V] 
wherein Xa is a substituted amino group and a process for production 
thereof are disclosed in J. Med. Chem., 14, 103 (1971). 
Compound [IV] is also known and may be obtained by a lithium aluminum 
hydride method (U.S. Pat. No. 3,738,998); hydrolysis by alklai (U.K. Pat. 
No. 1,250,063); a sodium alcoholate method [U.K. Pat. No. 1,250,063; J. 
Med. Chem., 14, 109 (1971)]; and by other like methods. 
A sodium alcoholate method which is illustrated in Reference Example 1 is 
briefly described below. 
Compound [IV] is obtained by eliminating carbamoyl from Compound [V] in the 
presence of an alcoholate in a solvent inert to the reaction. Suitable 
alcoholates for the reaction include alcoholates of methanol, ethanol, 
t-butanol and the like with sodium, potassium, and the like. Suitable 
solvents include methanol, ethanol, i-propanol, tetrahydrofuran, dioxane, 
dimethylformamide, benzene and the like. Preferably 5 to 7 times 
alcoholate is used per mole of Compound [V]. The reaction is usually 
carried out at room temperature and completed in several hours to several 
days. 
Process of Compound [IV].fwdarw.Compound [III] 
Compound [III] is obtained by reacting Compound [IV] with a reactive 
derivative of a carboxylic acid represented by the general formula 
EQU R.sup.1 COOH 
wherein R.sup.1 has the same meaning as defined before, such as an acid 
halide, an acid anhydride and the like (hereinafter referred to as an 
acylating agent), in the presence of a base in an inert solvent. 
Suitable acylating agents for the reaction include acetyl chloride, 
propionyl chloride, acetic anhydride and the like. Suitable bases include 
sodium carbonate, sodium hydride, triethylamine, pyridine and the like. 
Suitable solvents include tetrahydrofuran, dioxane, chloroform and the 
like. Certain bases, such as pyridine, may also function as the solvent. 
Preferably 1 to 2 times acylating agent is used per mole of Compound [IV]. 
Preferably 1 to 100 times base is used per mole of Compound [IV]. The 
reaction is usually carried out at -78.degree. to 30.degree. C. 
Process of Compound [III].fwdarw.Compound [II] 
Compound [II] is obtained by reacting Compound [III] with a reactive 
derivative of a compound represented by the general formula 
EQU R.sup.2 SO.sub.2 OH 
wherein R.sup.2 has the same meaning as defined before (hereinafter 
referred to as a sulfonylating agent), in the presence of a base in an 
inert solvent. 
Suitable sulfonylating agents for the reaction include halides, acid 
anhydrides and the like of the compound represented by the above general 
formula, such as methanesulfonyl chloride, trifluoromethanesulfonyl 
chloride, p-toluenesulfonyl chloride and the like. Suitable bases include 
inorganic bases such as sodium carbonate, sodium hydride and the like and 
organic bases such as triethylamine, pyridine and the like. These organic 
bases also function as the solvent. Suitable solvents include 
tetrahydrofuran, dioxane, chloroform and the like. 
Typically, 1 to 2 times sulfonylating agent is used per mole of Compound 
[III]. The base is typically used in an amount of 1 to 100 times per mole 
of Compound [III]. The reaction is usually carried out at -78.degree. to 
30.degree. C. 
Process of Compound [II].fwdarw.Compound [I-1] 
Compound [I-1] is obtained by eliminating alkyl (or aryl)-sulfonic acid 
from Compound [II] in the presence of a base in an inert solvent. 
Suitable bases for the reaction include inorganic bases such as sodium 
hydroxide, sodium hydride and the like and organic bases such as 
potassium-t-butoxide, 1,5-diazabicyclo[5.4.0]undecene-5, triethylamine, 
sodium methoxide, preferably potassium t-butoxide, 
1,5-diazabicyclo[5.4.0]-undecene-5 and the like. Suitable solvents include 
ethyl ether, tetrahydrofuran, dioxane, ethylene glycol dimethyl ether, 
n-hexane, petroleum ether, benzene, N,N-dimethylformamide, ethyl acetate, 
acetone, methylene chloride, dimethylsulfoxide and the like. 
The base is typically used in an amount of 1 to 20 times per mole of 
Compound [II]. The reaction is generally carried out at 0.degree. to 
80.degree. C. and is usually completed in one hour to one week. 
Process of Compound [I-1].fwdarw.Compound [I-2] 
Compound [I-2] is obtained by hydrolyzing Compound [I-1] in the presence of 
a base in water and an inert solvent. 
Suitable bases for the reaction include sodium hydrogen carbonate, ammonia, 
diethylamine, hydrazine and the like. Suitable inert solvents include 
methanol, ethanol, tetrahydrofuran, dimethylformamide, ethyl acetate, 
acetone, chloroform and the like. 
The base is typically used in an amount of 1 to 20 times per mole of 
Compound [I-1]. The reaction is generally carried out at 0.degree. to 
60.degree. C. and usually completed in 1 to 10 hours. 
Process of Compound [IV].fwdarw.Compound [I-3] 
Compound [I-3] is obtained by reacting Compound [IV] with a compound 
represented by the general formula 
EQU CH.sub.2 .dbd.CHZ 
wherein Z has the same meaning as defined before (hereinafter referred to 
as an alkylating agent) in an inert solvent to alkylate the 1a-N-position 
of Compound [IV]. 
When an alkylating agent active to the reaction (for example, acrolein) is 
used, the reaction proceeds simply by mixing the agent with Compound [IV]. 
Nevertheless, the reaction is usually carried out in the presence of a 
base such as triethylamine, sodium methoxide, potassium t-butoxide, and 
the like so that the reaction proceeds smoothly. Suitable solvents for the 
reaction include ethyl ether, tetrahydrofuran, dioxane, ethylene glycol 
dimethyl ether, n-hexane, petroleum ether, benzene, N,N-dimethylformamide, 
ethyl acetate, acetone, methylene chloride, chloroform, dimethylsulfoxide 
and the like. 
The alkylating agent is typically used in an amount of 1 to 30 times per 
mole of Compound [IV]. 
The reaction is carried out preferably at 20.degree. to 80.degree. C. and 
usually completed in one hour to one week. 
The alkylating method used in this process is known as a Michael addition 
and is disclosed in "Ethylenimine and Other Aziridines" 136, 1969 
(Academic Press, U.S.A.). 
Process of Compound [I-3].fwdarw.Compound [I-4] 
Compound [I-4] is obtained in the same manner as described in the process 
of Compound [III].fwdarw.Compound [II]. 
Process of Compound [I-4].fwdarw.Compound [I-5] 
Compound [I-5] is obtained in the same manner as described in the process 
of Compound [II].fwdarw.Compound [I-1]. 
Process of Compound [I-5].fwdarw.Compound [I-2] 
Compound [I-2] is obtained by subjecting Compound [I-5] to retro Michael 
reaction in the presence of an acid or a base in an inert solvent. 
Suitable acids for the reaction include inorganic acids such as perchloric 
acid, and the like. Suitable bases include tertiary amines such as 
N,N-dimethylaniline, and the like. Suitable solvents include ethyl ether, 
tetrahydrofuran, ethylene glycol dimethyl ether, n-hexane, pertroleum 
ether, benzene, N,N-dimethylformamide, ethyl acetate, acetone, methylene 
chloride, chloroform, dimethylsulfoxide and the like. 
The acid or base is typically used in an amount of 1 to 100 times per mole 
of Compound [I-5]. 
The reaction is preferably carried out at 0.degree. to 80.degree. C. and 
usually completed in 10 minutes to several hours. 
Retro Michael reaction is disclosed in Tetrahedron Letters, No. 49, 4295 
(1977), Synthesis, No. 12, 745 (1973). 
Process of Compound [I-4].fwdarw.Compound [I-6] 
Compound [I-6] is obtained in the same manner as described in the process 
of Compound [I-5].fwdarw.Compound [I-2]. 
Process of Compound [I-6].fwdarw.Compound [I-2] 
Compound [I-2] is obtained in the same manner as described in the process 
of Compound [II].fwdarw.Compound [I-1]. 
Process of Compound [I-2-1].fwdarw.Compound [I-7] 
Compound [I-7] is obtained by hydrolizing Compound [I-2-1] in an aqueous 
solution of a base. 
Suitable bases include inorganic bases such as sodium hydroxide, sodium 
carbonate and the like and organic bases such as triethylamine and the 
like. 
The base is usually used in a concentration of 0.01 to 1 normality. The 
reaction is usually carried out at room temperature and completed in 30 
minutes to several hours. 
Process of Compound [I-7].fwdarw.Compound [I-2-2] 
Compound [I-2-2] is obtained by reacting Compound [I-7] with an alkylating 
agent in an inert solvent. 
Suitable alkylating agents for the reaction include diazoalkanes such as 
diazomethane and the like, alkyl halides such as methyl iodide and the 
like, dialkyl sulfates such as dimethyl sulfate and the like. Suitable 
solvents include ethyl acetate, ethyl ether and the like. When an acid is 
produced by the reaction, the reaction is carried out in the presence of 
an acid acceptor such as potassium carbonate, triethylamine and the like. 
Process of Compound [I-2-2].fwdarw.Compound [I-2-1] 
Compound [I-2-1] is obtained by reacting Compound [I-2-2] with ammonia or 
an amine in an inert solvent such as methanol and the like. 
The ammonia or amine is typically used in an amount of 1 to 100 times per 
mole of Compound [I-2-2]. The reaction is usually carried out at 
20.degree. to 80.degree. C. and completed in 1 to several hours. 
Process of Compound [VI].fwdarw.Compound [I-8] 
Compound [I-8] is obtained in the same manner as described in the process 
of Compound [I-2-1].fwdarw.Compound [I-7]. 
Although Compound [VI] and preparation thereof are disclosed in U.S. Patent 
Application Ser. No. 58,670 filed July 18, 1979, the preparation is 
illustrated in Reference Example 4. 
In each of the above processes, recovery of the desired compound from the 
reaction mixture is carried out by conventional methods such as those 
described in the following Examples and Reference Examples.

Certain specific embodiments of the invention are illustrated by the 
following representative examples. 
EXAMPLE 1 
Preparation of Compound [I-1] from Compound [II] 
In this example, 38 mg of 
1a-N-acetyl-10-decarbamoyl-10-methanesulfonyl-mitomycin C obtained in 
Reference Example 3 is dissolved in 8 ml of ethylene glycol dimethyl 
ether. Then, 150 mg of 1,5-diazabicyclo[5.4.0]undecene-5 is added to the 
solution and the mixture is refluxed with heating for 2 hours in an 
atmosphere of nitrogen. The reaction mixture is then poured in saturated 
aqueous solution of sodium hydrogen carbonate and the mixture is extracted 
with ethyl acetate. The extract is washed with water, dried with anhydrous 
sodium sulfate and concentrated under reduced pressure to remove the 
solvent. The residue is then purified by silica gel column chromatography 
using a mixed solvent of chloroform and acetone (6:4) (volume ratio, as is 
the same hereinafter) as a developer to obtain 24 mg of deep green 
crystals having the following physical properties: 
Mass spectrum: 
The substance exhibits molecular ion peak at m/e 315 
.sup.1 HNMR spectrum (in pyridine-d.sub.5, .delta. (ppm)): 
2.03(s,3H), 2.05(s,3H), 3.17(s,3H), 3.57(dd,1H), 3.64(dd,1H), 3.78(d,1H), 
4.84(d,1H), 5.61(d,1H), 6.55(d,1H), 7.76(bs,2H) 
IR spectrum (KBr tablet, cm.sup.-1): 
3435(w), 3330(m) (N-H stretch), 1695(s), 1594(vs), 1537(vs) (C.dbd.O 
stretch), 1656(m) (C.dbd.C stretch) 
From the above properties, the substance is identified as 
1a-N-acetyl-10-decarbamoyloxy-9-dehydro-mitomycin C. Yield 82%. 
EXAMPLE 2 
Preparation of Compound [I-2] from Compound [I-1] 
In this example, 57 mg of 1a-N-acetyl-10-decarbamoyloxy-9-dehydro-mitomycin 
C is dissolved in 1 ml of methanol. Then, 1 ml of 10% aqueous solution of 
sodium hydrogen carbonate is added to the solution and the mixture is 
stirred at room temperature for 6 hours. After the completion of the 
reaction, the reaction mixture is extracted with ethyl acetate. The 
extract is then dried with anhydrous sodium sulfate and concentrated under 
reduced pressure. 
The residue is purified by silica gel column chromatography using a mixed 
solvent of chloroform and acetone (1:1) to obtain 36 mg of deep green 
crystals having the following physical properties: Mass spectrum: 
The substance exhibits molecular ion peak at m/e 273. Molecular weight 
obtained by high resolution mass spectrometry is 273.1130 (273.1113 as 
calculated). 
.sup.1 HNMR spectrum (in pyridine-d.sub.5, .delta. (ppm)): 2.00(s,3H), 
2.77(dd,1H), 3.04(d,1H), 3.17(s,3H), 3.59(dd,1H), 4.67(d,1H), 5.50(d,1H), 
6.50(d,1H), 7.57(bs,2H) 
IR spectrum (KBr tablet, cm.sup.-1) (FIG. 1): 3420(m) (N-H stretch), 
3320(m) (N-H) stretch), 3285(m) (N-H stretch), 1651(m) (C.dbd.C stretch), 
1592(vs) (C.dbd.O stretch), 1532(vs) (C.dbd.O stretch) 
From the foregoing properties, the substance is identified as 
10-decarbamoyloxy-9-dehydro-mitomycin C. Yield 73%. 
EXAMPLE 3 
Preparation of Compound [I-3] from Compound [IV] 
In this example, 66 mg of 10-decarbamoyl-mitomycin C obtained in Reference 
Example 1 is dissolved in 5 ml of methylene chloride. Then, 0.3 ml of 
acrolein (purity 90%) is added to the solution. The mixture is then 
stirred at room temperature for 4 days and concentrated under reduced 
pressure. The residue is purified by silica gel column chromatography 
using a mixed solvent of chloroform and methanol (93:7) to obtain 76 mg of 
purplish black crystals having the following physical properties: 
.sup.1 HNMR spectrum (in pyridine-d.sub.5, .delta. (ppm)): 2.00(s,3H), 
2.39-2.96(m,4H), 2.40(dd,1H), 2.91(d,1H) 3.22(s,3H), 3.61(dd,1H), 
3.83(dd,1H), 4.34(dd,1H), 4.49(d,1H), 4.75(dd,1H), 6.18(bs,1H), 
7.58(bs,2H), 9.76(t,1H) 
IR spectrum (KBr tablet, cm.sup.-1): 3430(m) (N-H stretch), 3335(m) (N-H 
stretch), 1722(m) (C.dbd.O stretch), 1604(s) (C.dbd.O stretch), 1554(vs) 
(C.dbd.O stretch) 
From the foregoing properties, the substance is identified as 
10-decarbamoyl-1a-N-(2-formylethyl)-mitomycin C. Yield 97%. 
EXAMPLE 4 
Preparation of Compound [I-4] from Compound [I-3] 
In this example, 62 mg of 10-decarbamoyl-1a-N-(2-formylethyl)-mitomycin C 
obtained in Example 3 is dissolved in 0.33 ml of anhydrous pyridine. Then, 
15 .mu.of methanesulfonyl chloride is added to the solution in an 
atmosphere of nitrogen and the mixture is stirred under ice cooling and 
sodium chloride for 2 hours. The reaction mixture is then poured into a 
saturated aqueous solution of sodium hydrogen carbonate and the mixture is 
extracted with chloroform. The extract is then washed with water, dried 
with anhydrous sodium sulfate and concentrated under reduced pressure. The 
residue is purified by silica gel column chromatography using a mixed 
solvent of chloroform and acetone (6:4) to obtain 44 mg of brownish black 
crystals having the following physical properties: 
.sup.1 HNMR spectrum (in pyridine-d.sub.5, .delta. ppm): 1.99(s,3H), 
2.17-2.35(m,2H), 2.46(dd,1H), 2.64(dt,2H), 2.84(d,1H), 3.20(s,3H), 
3.35(s,3H), 3.59(dd,1H), 4.01(dd,1H), 4.47(d,1H), 4.88(dd,1H), 
5.37(dd,1H), 7.66(bs,2H), 9.75(t.1H) 
IR spectrum (KBr tablet, cm.sup.-1): 3440(m) (N-H stretch), 3340(m) (N-H 
stretch), 1720(m) (C.dbd.O stretch), 1604(vs) (C.dbd.O stretch), 1555(vs) 
(C.dbd.O stretch), 1351(vs) (antisym. SO.sub.2, stretch), 1174(vs) (sym. 
SO.sub.2, stretch) 
From the foregoing properties, the substance is identified as 
10-decarbamoyl-1a-N-(2-formylethyl)-10-methansulfonyl-mitomycin C. Yield 
58%. 
EXAMPLE 5 
Preparation of Compound [I-5] from Compound [I-4] 
In this example, 52 mg of 
10-decarbamoyl-1a-N-(2-formylethyl)-10-methanesulfonyl-mitomycin C 
obtained in Example 4 is dissolved in 10 ml of anhydrous ethylene glycol 
dimethyl ether. Then, 210 mg of 1,5-diazabicylo [5.4.0] undecene-5 is 
added to the solution and the mixture is refluxed in an atmosphere of 
nitrogen for 2 hours. The reaction mixture is then poured into a saturated 
aqueous solution of sodium hydrogen carbonate and the mixture is extracted 
with ethyl acetate. The extract is washed with water, dried with anhydrous 
sodium sulfate and concentrated under reduced pressure. The residue is 
purified by silica gel column chromatography using a mixed solvent of 
chloroform and acetone (6:4) to obtain 16 mg of deep green crystals having 
the following physical properties: 
.sup.1 HNMR spectrum (in pyridine-d.sub.5, .delta. (ppm)): 2.00(s,3H), 
2.29-2.78(m,4H), 2.40(dd,1H), 2.66(d,1H), 3.14(s,3H), 3.53(dd,1H), 
4.64(d,1H), 5.51(d,1H), 6.51(d,1H), 7.68(bs,2H), 9.71(t,1H) 
IR spectrum (KBr tablet, cm.sup.-1): 3420(m) (N-H stretch), 3225(m) (N-H 
stretch), 1718(m) (C.dbd.O stretch), 1649(m) (C.dbd.C stretch), 1592(vs) 
(C.dbd.O stretch), 1535(vs) (C.dbd.O stretch) 
From the foregoing properties, the substance is identified as 
10-decarbamoyloxy-9-dehydro-1a-N-(2-formylethyl)mitomycin C. Yield 40%. 
EXAMPLE 6 
Preparation of Compound [I-2] from Compound [I-5] 
In this example, 26 mg of 
10-decarbamoyloxy-9-dehydro-1a-N-(2-formylethyl)-mitomycin C obtained in 
Example 5 is dissolved in 4 ml of anhydrous methylene chloride. Thereupon, 
first 100 mg of a salt of N,N-dimethylaniline with perchloric acid and 
subsequently 0.4 ml of N,N-dimethylaniline are added to the solution. The 
mixture is stirred at room temperature for 15 minutes and poured into a 
saturated aqueous solution of sodium hydrogen carbonate. The mixture is 
then extracted with chloroform. The extract is washed with water, dried 
with anhydrous sodium sulfate and concentrated under reduced pressure. The 
residue is purified by silica gel column chromatography using a mixed 
solvent of chloroform and acetone (1:1) to obtain 15 mg of deep green 
crystals. 
Mass spectrum, .sup.1 HNMR spectrum and IR spectrum of the substance are in 
accord with those of the compound obtained in Example 2. Therefore, the 
substance is identified as 10-decarbamoyloxy-9-dehydro-mitomycin C. Yield 
70%. 
EXAMPLE 7 
Preparation of Compound [I-6] from Compound [I-4 
In this example, 33 mg of 
10-decarbamoyl-1a-N-(2-formylethyl)-10-methanesulfonyl-mitomycin C 
obtained in Example 4 is dissolved in 5 ml of anhydrous methylene 
chloride. Thereupon, first 120 mg of a salt of N,N-dimethylaniline with 
perchloric acid and subsequently 0.5 ml of N,N-dimethylaniline are added 
to the solution. The mixture is stirred at room temperature for 30 
minutes. The reaction mixture is poured into a saturated aqueous solution 
of sodium hydrogen carbonate and the mixture is extracted with chloroform. 
The extract is then washed with water, dried with anhydrous sodium sulfate 
and concentrated under reduced pressure. The residue is purified by silica 
gel column chromatography using a mixed solvent of chloroform and methanol 
(97:3) to obtain 21 mg of purplish red crystals having the following 
physical properties: 
.sup.1 HNMR spectrum (in pyridine-d.sub.5, .delta.(ppm)): 2.04(s,3H), 
2.82(dd,1H), 3.22(s,3H), 3.23(d,1H), 3.63 (dd,1H), 4.02(dd,1H), 
4.52(d,1H), 5.18(dd,1H), 5.32(dd,1H), 7.66(bs,2H) 
IR spectrum (KBr tablet, cm.sup.-1): 3440(w) (N-H stretch), 3315(m)(N-H 
stretch), 1604(vs) (C.dbd.O stretch), 1555(vs)(C.dbd.O stretch), 1352(vs) 
(antisym. SO.sub.2, stretch), 1174(vs) (sym. SO.sub.2, stretch) 
From the foregoing properties, the substance is identified as 
10-decarbamoyl-10-methanesulfonyl-mitomycin C. Yield 72%. 
EXAMPLE 8 
Preparation of Compound [I-2] from Compound [I-6 ] 
In this example, 20.5 mg of 10-decarbamoyl-10-methanesulfonyl-mitomycin C 
obtained in Example 7 is dissolved in 0.3 ml of anhydrous tetrahydrofuran. 
Then, 30 mg of 1,5-diazabicyclo[5.4.0] undecene-5 is added to the solution 
and the mixture is stirred at room temperature for 40 hours. The reaction 
mixture is then poured into a saturated aqueous solution of sodium 
hydrogen carbonate and the mixture is extracted with ethyl acetate. The 
extract is washed with water, dried with anhydrous sodium sulfate and 
concentrated under reduced pressure. The residue is then purified by 
silica gel column chromatography using a mixed solvent of chloroform and 
acetone (1:1) to obtain 1.2 mg of deep green crystals. 
Mass spectrum, .sup.1 HNMR spectrum and IR spectrum of the substance are in 
accord with those of the compound obtained in Example 2. Therefore, the 
substance is identified as 10-decarbamoyloxy-9-dehydro-mitomycin C. Yield 
7.9%. 
EXAMPLE 9 
Preparation of Compound [I-7] from Compound [I-2-1] 
In this example, 18 mg of 10-carbamoyloxy-9-dehydro-mitomycin C obtained in 
Example 2 is dissolved in 3.75 ml of 0.1N sodium hydroxide. The solution 
is stirred at room temperature for 45 minutes. The reaction solution is 
then adjusted to pH 4 with diluted hydrochloric acid and extracted with 
ethyl acetate. The extract is washed with water, dried with anhydrous 
sodium sulfate and concentrated under reduced pressure. The residue is 
then purified by silica gel column chromatography using a mixed solvent of 
chloroform and methanol (9:1) to obtain 12 mg of purplish black crystals 
having the following physical properties: 
Mass spectrum: 
The substance exhibits molecular ion peak at m/e 274. 
.sup.1 HNMR spectrum (in pyridine-d.sub.5, .delta.(ppm)): 2.05(s,3H), 
2.80(dd,1H), 3.08(d,1H), 3.19(s,3H), 3.58(dd, 1H), 4.50(d,1H), 5.55(d,1H), 
6.52(d,1H) 
IR spectrum (KBr tablet, cm.sup.-1), (FIG. 2): 3285(w) (N-H stretch, 
1644(vs) (C.dbd.C stretch), 1630(vs) (C.dbd.O stretch), 1548(C.dbd.O 
stretch) 
From the foregoing properties, the substance is identified as 
7-deamino-10-decarbamoyloxy-9-dehydro-7-hydroxy-mitomycin C. Yield 66%. 
EXAMPLE 10 
Preparation of Compound [I-2-2] from Compound [I-7 
In this example, 12 mg of 
7-deamino-10-decarbamoyloxy-9-dehydro-7-hydroxy-mitomycin C obtained in 
Example 9 is dissolved in 3 ml of ethyl acetate. An excess amount of ethyl 
ether solution of diazomethane is added dropwise to the solution under ice 
cooling and the mixture is allowed to stand for 10 minutes. The mixture is 
then concentrated under reduced pressure and the reside is purified by 
silica gel column chromatography using a mixed solvent of chloroform and 
acetone (6:4) to obtain 8 mg of purplish black crystals having the 
following physical properties: 
Mass spectrum: 
The substance exhibits molecular ion peak at m/e 288. Molecular weight 
obtained by high resolution mass spectrometry is 288.1120 (288.1110 as 
calculated). 
.sup.1 HNMR spectrum (in pyridine-d.sub.5, .delta.(ppm)): 1.83(dd,1H), 
3.07(d,1H), 3.16(s,3H), 3.53(dd,1H), 4.01(s,3H), 4.33(d,1H), 5.59(d,1H), 
6.57(d,1H) 
IR spectrum (KBr tablet, cm.sup.-1), (FIG. 3): 3305(vw) (N-H stretch), 
1650(vs) (C.dbd.O stretch), 1554(vs) (C.dbd.O stretch). 
From the foregoing properties, the substance is identified as 
10-decarbamoyloxy-9-dehydro-mitomycin A. Yield 63%. 
EXAMPLE 11 
Preparation of Compound [I-2-1] from Compound [I-2-2] 
In this example, 5.2 mg of 10-decarbamoyloxy-9-dehydro-mitomycin A obtained 
in Example 10 is dissolved in 2 ml of methanol saturated with ammonia. The 
solution is stirred at room temperature for 2 hours. The solution is then 
concentrated under reduced pressure and the residue is purified by silica 
gel column chromatography using a mixed solvent of chloroform and acetone 
(1:1) to obtain 4.1 mg of deep green crystals. 
Mass spectrum, .sup.1 HNMR spectrum and IR spectrum of the substance are in 
accord with those of the compound obtained in Example 2. Therefore, the 
substance is identified as 10-decarbamoyloxy-9-dehydro-mitomycin C. Yield 
83%. 
EXAMPLE 12 
Preparation of Compound [I-8] from Compound [VI] 
In this example, 134 mg of 10-decarbamoyloxy-9-dehydro-porfiromycin 
obtained in Reference Example 4 is dissolved in 25 ml of 0.1N sodium 
hydroxide. The solution is stirred at room temperature for 4 hours. The 
reaction solution is adjusted to pH 3 with diluted hydrochloric acid and 
extracted with ethyl acetate. The extract is washed with water, dried with 
anhydrous magnesium sulfate and concentrated under reduced pressure. The 
residue is purified by silica gel column chromatography using a mixed 
solvent of chloroform and methanol (9:1) and crystallized from a mixed 
solvent of chloroform and methanol to obtain 115 mg of dark green needle 
crystals having the following physical properties: 
Mass spectrum: 
The substance exhibits molecular ion peak at m/e 288. 
.sup.1 HNMR spectrum (in pyridine-d.sub.5, .delta.(ppm)): 2.03(s,3H), 
2.12(s,3H), 2.22(dd,1H), 2.48(d,1H), 3.13 (s,3H), 3.47(dd,1H), 4.44(d,1H), 
5.44(bs,1H), 6.48(bs,1H) 
IR spectrum: 
IR spectrum (KBr tablet) is shown in FIG. 4. 
From the foregoing properties, the substance is identified as 
10-decarbamoyloxy-9-dehydro-7-deamino-7-hydroxy-porfiromycin. Yield 85.3%. 
REFERENCE EXAMPLE 1 
Preparation of Compound [IV] from Compound [V] 
In this example, 230 mg of sodium is dissolved in 50 ml of isopropanol and 
500 mg of mitomycin C is added to the solution. The mixture is stirred at 
room temperature for 8 hours. The reaction mixture is then neutralized 
with an excess amount of dry ice. The precipitate is filtered out and the 
filtrate is concentrated under reduced pressure. The residue is purified 
by silica gel column chromatography using a mixed solvent of chloroform 
and methanol (9:1) to obtain 270 mg of purplish black crystals. 
The substance is identified as the known compound 10-decarbamoyl-mitomycin 
C from melting point, .sup.1 HNMR spectrum, TLC, and similar properties. 
Yield 62%. 
REFERENCE EXAMPLE 2 
Preparation of Compound [III] from Compound [IV] 
In this example, 138 mg of 10-decarbamoyl-mitomycin C obtained in Reference 
Example 1 is dissolved in 1 ml of pyridine. Then, 60 .mu.l of acetic 
anhydride is added to the solution under ice cooling and sodium chloride 
and the mixture is stirred in an atmosphere of nitrogen for 1 hour. The 
reaction mixture is then poured into a saturated aqueous solution of 
sodium hydrogen carbonate and the mixture is extracted with ethyl acetate. 
The extract is washed with water, dried with anhydrous sodium sulfate and 
concentrated under reduced pressure. The residue is then purified by 
silica gel column chromatography using a mixed solvent of chloroform and 
methanol (95:5) to obtain 153 mg of purplish black crystals having the 
following physical properties: Mass spectrum: 
The substance exhibits molecular ion peak at m/e 333. .sup.1 NHMR spectrum 
(in pyridine-d.sub.5, .delta.(ppm): 2.05(s,3H), 2.23(s,3H), 3.23(s,3H), 
3.57(dd,1H), 3.66(dd,1H), 3.94(dd,1H), 3.98(d,1H), 4.33(dd,1H), 
4.76(d,1H), 4.84(dd,1H), 7.57(bs,2H) 
IR spectrum (KBr tablet, cm.sup.-1): 3435(m), 3335(m) (N-H stretch), 
1694(m), 1605(s), 1555(vs) (C.dbd.O stretch) 
From the foregoing properties, the substance is identified as 
1a-N-acetyl-10-decarbamoyl-mitomycin C. Yield 97%. 
REFERENCE EXAMPLE 3 
Preparation of Compound [II] from Compound [III] 
In this example, 62 mg of 1a-N-acetyl-10-decarbamoylmitomycin C obtained in 
Reference Example 2 is dissolved in 0.5 ml of anhydrous pyridine. Then, 15 
.mu.l of methanesulfonyl chloride is added to the solution in an 
atmosphere of nitrogen under ice cooling and sodium chloride and the 
mixture is stirred for 2 hours. The reaction mixture is poured in a 
saturated aqueous solution of sodium hydrogen carbonate and the mixture is 
extracted with ethyl acetate. The extract is washed with water, dried with 
anhydrous sodium sulfate and concentrated under reduced pressure. The 
residue is purified by silica gel column chromatography using a mixed 
solvent of chloroform and methanol (96:4) to obtain 74 mg of purplish 
black crystals having the following physical properties: 
.sup.1 HNMR spectrum (in pyridine-d.sub.5, .delta.(ppm): 2.04(s,3H), 
2.11(s,3H), 3.20(s,3H), 3.46(s,3H), 3.60(dd,1H), 3.61(dd,1H), 3.79(d,1H), 
4.09(dd,1H), 4.84(dd,1H), 5.58(dd,1H), 7.71(bs,2H) 
IR spectrum (KBr tablet, cm.sup.-1): 3445(w), 3235(m) (N-H stretch), 
1697(s), 1605(s), 1563(vs) (C.dbd.O stretch), 1350(vs) (antisym. SO.sub.2 
stretch), 1173(vs) (sym. SO.sub.2 stretch) 
From the foregoing properties, the substance is identified as 
1a-N-acetyl-10-decarbamoyl-10-methanesulfonylmitomycin C. Yield 97%. 
REFERENCE EXAMPLE 4 
Synthesis of Compound [VI] 
In this example, 500 mg of porfiromycin is added to 150 ml of isopropanol 
containing 1.5 g of sodium isopropoxide. The mixture is stirred at room 
temperature for 6 hours. The reaction mixture is neutralized with an 
excess amount of dry ice and the deposit is filtered out. The filtrate is 
then concentrated under reduced pressure and the residue is purified by 
silica gel column chromatography using a mixed solvent of chloroform and 
methanol (94:6) to obtain 299 mg of purplish blue crystals having the 
following physical properties: 
.sup.1 HNMR spectrum (in CD.sub.3 OD, .delta.(ppm)): 1.75(s,3H), 
2.29(s,3H), 2.45(dd,1H), 2.54(d,1H), 3.20(s,3H), 3.34(dd,1H, 3.46(dd,1H), 
3.80(dd,1H), 4.09(dd,1H), 4.16(d,1H) 
From the foregoing properties, the substance is identified as 
10-decarbamoyl-porfiromycin. Yield 68.2%. 
Thereupon, 105.4 mg of 10-decarbamoyl-porfiromycin is dissolved in 2 ml of 
anhydrous pyridine. Then, 0.05 ml of methanesulfonyl chloride is added to 
the solution and the mixture is stirred for 20 minutes. The reaction 
mixture is poured into 10 ml of a saturated aqeuous solution of sodium 
hydrogen carbonate and the mixture is extracted with ethyl acetate. The 
extract is washed with water, dried with anhydrous sodium sulfate and 
concentrated under reduced pressure. The residue is purified by silica gel 
column chromatography using a mixed solvent of chloroform and methanol 
(95:5) to obtain 130.7 mg of purple solid having the following physical 
properties: Mass spectrum: 
The substance exhibits molecular ion peak at m/e 383. 
.sup.1 HNMR spectrum (in CD.sub.3 OD, .delta.(ppm)): 
1.74(s,3H), 2.33(s,3H), 2.45(dd,1H), 2.53(d,1H), 3.18(s,3H), 3.22(s,3H), 
3.46(dd,1H), 3.62(dd,1H), 4.19(d,1H), 4.42(dd,1H), 4.82(dd,1H) 
From the foregoing properties, the substance is identified as 
10-decarbamoyl-10-methanesulfonyl-porfiromycin. Yield 98.6%. 
Thereafter, 17 mg of 10-decarbamoyl-10-methanesulfonyl-porfiromycin, as 
thus obtained is dissolved in 1 ml of anhydrous tetrahydrofuran. Then, 54 
mg of 1,5-diazabicyclo[5.4.0]undecene-5 is added to the solution and the 
mixture is refluxed in an atmosphere of nitrogen for 5 hours. The reaction 
mixture is then concentrated under reduced pressure and the residue is 
purified by silica gel column chromatography using a mixed solvent of 
chloroform and acetone (4:1) to obtain 7.5 mg of purplish blue crystals 
having the following physical properties: 
Mass spectrum: 
The substance exhibits molecular ion peak at m/e 287. 
.sup.1 HNMR spectrum (in CD.sub.3 OD, .delta.(ppm)): 1.78(s,3H), 
2.21(s,3H), 2.44(bs,2H), 3.06(s,3H), 3.42(dd,1H, 4.26(d,1H), 5.34(d,1H), 
6.08(d,1H) 
From the above properties, the substance is identified as 
10-decarbamoyloxy-9-dehydro-porfiromycin. Yield 58.9%.