JPS626556B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to novel mitomycins and methods for producing the same. Mitomycins are generally known as substances having antitumor activity, and representative compounds thereof include mitomycin A, B, C, and porphyromycin (The Merck Index 9th Ed.). The present applicant has also proposed mitomycin D (patent application 1983-29238) and mitomycin E (patent application 1983-29238).
29238), mitomycin F (patent application 1982-117836)
An application has been filed for an invention related to this invention. These mitomycins have the following structure.

[Table] Various derivatives of these compounds are also known. For example, 1a-N-acetylmitomycin C, 1a-N-butyrylmitomycin C (the above,
Special Publication No. 40-18117, U.S. Patent No. 3514452), 7-
Ethylamino-7-demethoxymitomycin A
(Special Publication No. 40-20718, U.S. Patent No. 3514452), 10-
Decarbamoyl mitomycin A, B and C
(Special Publication No. 46-31229, U.S. Patent No. 3738998), 1a-
N-acetyl-10-decarbamoyl mitomycin C (Special Publication No. 49-17279), 1a-N-acetyl-10
-decarbamoyl-10-paratoluenesulfonyl mitomycin C, 10-decarbamoyl-10-paratoluenesulfonylporphyromycin (Japanese Patent Publication No. 37639/1989), and the like. Recently, a series of mitomycins having double bonds at the 9 and 10 positions and having excellent antibacterial activity have been discovered, as shown in the following formula (Japanese Patent Application No. 86,748/1986). (In the formula, X _B represents NH ₂ or alkoxy, and Y
_A represents H or an alkyl group). Compounds with excellent properties are always in demand, and as a result of our studies on the synthesis of new mitomycins, the present inventors found mitomycins represented by the following general formula [] (hereinafter sometimes referred to as compounds []). The same applies to compounds with other formula numbers. ] was found to be a new compound with antibacterial properties, and the present invention was completed. [In the formula, X is amino, substituted amino, hydroxy or alkoxy, Y is hydrogen, -COR ₁ (in the formula, R ₁
represents hydrogen, alkyl, substituted alkyl, aryl or substituted aryl) or -CH ₂ CH ₂ Z
(In the formula _, Z represents alkyloxycarbonyl, acyl or cyano), R _A and R _B are hydrogen and R _B is -CH ₂ OH or -CH ₂ OSO ₂ R ₂ (in the formula,
R ₂ represents alkyl, substituted alkyl or aryl), or R _A and R _B taken together represent =CH ₂ . However, if R _A is hydrogen and R _B is -CH ₂ OH, then Y is -CH ₂ CH ₂ Z, and if R _A is hydrogen and R _B is -CH ₂ OSO ₂ R ₂ , Y is Hydrogen or -
Suppose that CH ₂ CH ₂ Z. ]. The object compound (compound []) of the present invention has a wide range of antibacterial activity. Therefore, it is useful for sterilizing instruments, etc. in hospitals, as a cleaning agent together with detergents, and as a medicine. Compound [ ] is also useful as an intermediate for creating new useful compounds. Next, the present invention will be explained in more detail. In the group X of the compound [], examples of alkoxy include alkoxy having 1 to 4 carbon atoms, such as methoxy, ethoxy, i-propoxy, and n-butoxy. In the group _R1 , the alkyl is an alkyl having 1 to 3 carbon atoms, such as methyl, ethyl, i-propyl, etc., the substituted alkyl is, for example, chloromethyl, the aryl is, for example, phenyl, and the substituted aryl is, for example, p-methoxyphenyl. can be given. In the group Z, the alkyl of alkyloxycarbonyl is an alkyl having 1 to 4 carbon atoms, such as methyl, ethyl, t-butyl, etc., and the acyl is -COR ₄ [wherein R ₄ is hydrogen or an alkyl having 1 to 3 carbon atoms] represents alkyl, such as methyl, ethyl, isopropyl, etc.]. Suitable radicals R ₂ include, for example, methyl, trifluoromethyl, p-methylphenyl, and the like. Representative compounds are shown below.

【table】

[Table] Hereinafter, the compounds with compound numbers 1, 2, etc. may be referred to as compounds 1, 2, etc. The physical property values of these compounds are as shown in each example. Minimum inhibitory concentration (μg/ml) of compounds 1 to 8 against various bacteria shown below (agar dilution method, PH
7.0) are shown in Tables 1 to 3. A. Serratia marcetuscens
marcescens) ATCC4003 B Pseudomonas cepacia (Pseudomonas
cepacia) ATCC 25608 C Staphylococcus aureus ATCC 6538P D Escherichia coli
ATCC 26 E Bacillus subtilis
#10707 F Proteus vulgaris
vulgalis) ATCC 6897 G Shigella sonnei
ATCC9290 H Salmonella taihosa (Salmonella
typhosa) ATCC 9992 I Klebsiella pneumoniae
pneu-moniae) ATCC 10031

【table】

【table】

【table】

[Table] Next, the method for producing compound [] will be explained.
An example of a manufacturing process diagram for compound [] is shown below. In the above process diagram, Xa represents amino, substituted amino or alkoxy (groups other than hydroxy in X). Xaa is amino,
Represents substituted amino (a group excluding the case where a is alkoxy). Xab represents alkoxy (a group other than amino or substituted amino in Xa). Z, R ₁ and R ₂ have the same meanings as above. Compounds [-1] to [-7] and [-2]
-1] and [-2-2] are both included in the compound []. Next, each of the above steps will be explained. Process of compound []→[] In compound [], if Xa=NH ₂ , it is mitomycin C, and if Xa=OCH ₃ , it is mitomycin A. Compounds [] in which Xa is alkoxy (excluding the case of OCH ₃ ) and their production methods are described in Japanese Patent Publication No. 42-22380, U.S. Patent No. 3558651, and Japanese Patent Publication No. 54-28250, when Xa is substituted amino. The compound [] and its production method are described in J.Med.Chem., 14 ,
103 (1971). Compound [] also has known compounds, and compound []
The lithium aluminum hydride method (Japanese Patent Publication No. 46-31229, U.S. Patent No. 3738998
), alkaline hydrolysis method (Japanese Patent Publication No. 47-24558, British Patent No. 1250063), sodium alcoholate method [Japanese Patent Publication No. 47-24560, British Patent No. 1250063, J.
Med.Chem., 64 109 (1971)]. The sodium alcoholate method shown in Reference Example 1 below will be briefly explained below. Compound [] in methanol, ethanol, t-
The compound [] is produced by hydrolysis with an alcoholate such as butyl alcohol. As the solvent, methanol, ethanol, isopropanol, tetrahydrofuran, dioxane, dimethylformamide, benzene, etc. are used. The appropriate amount of alcoholate to be used is 5 to 7 times the mole of compound []. The reaction generally proceeds at room temperature and is usually completed within several hours to several days. Process of compound []→compound [] Compound [] is a compound [] and a reactive derivative of a carboxylic acid represented by R ₁ COOH (wherein R ₁ has the same meaning as above), such as an acid halide, an acid anhydride. (hereinafter referred to as an acylating agent) in an inert solvent in the presence of a base. Although this method has already been reported (Japanese Patent Publication No. 17279/1983), it will be briefly explained below. As the acylating agent, acetyl chloride, propionyl chloride, acetic anhydride, etc. are used.
As the base, sodium carbonate, sodium hydride, triethylamine, pyridine, etc. are used. Tetrahydrofuran, dioxane, chloroform, etc. are used as the solvent, but a base may also serve as the solvent. It is appropriate to use the acylating agent in a molar amount of 1 to 2 times the amount of the compound [], and the base to be used in a molar amount of 1 to 100 times the amount of the compound []. A suitable reaction temperature is -78°C to 30°C. Process of compound [] → compound [] Compound [] is a compound _{[ ]} and a reactive derivative (hereinafter referred to as _sulfonylated agent) in an inert solvent in the presence of a base. Examples of the sulfonylating agent include halides and acid anhydrides of compounds of the above general formula, such as P-toluenesulfonic acid chloride, methanesulfonic acid chloride,
Trifluoromethanesulfonic anhydride and the like are used. As the base, inorganic bases (eg, sodium carbonate, sodium hydride, etc.) and organic bases (eg, triethylamine, pyridine, etc.) are used. These organic bases may also serve as a solvent. Tetrahydrofuran, dioxane, chloroform, etc. are used as the solvent. The sulfonylating agent is 1 to 2
The amount of base used is about 1 to 100 times the molar amount of the compound []. The reaction is suitably carried out at -78 to 30°C. Step of Compound [] → Compound [-1] Compound [-1] can be obtained by dealkylating (or aryl) sulfonating compound [] in the presence of a base in a solvent inert to the reaction. Bases include inorganic bases such as sodium hydroxide, sodium hydroxide, etc., and organic bases such as potassium tert-butoxide, 1.5-
Diazabicyclo [5.4.0] undecene-5,
Triethylamine, sodium methoxide, especially sodium t-butoxide, 1,5-diazabicyclo[5.4.0]undecene-5, etc. are used. Examples of solvents include ethers (e.g. ethyl ether, tetrahydrofuran, dioxane, ethylene glycol dimethyl ether, etc.), hydrocarbons (e.g. n-hexane, petroleum ether, benzene, etc.), amides (N/N-dimethylformamide, etc.), and esters. (ethyl acetate, etc.), ketones (acetone, etc.), halogenated hydrocarbons (methylene chloride, etc.), dimethyl sulfoxide, etc. are used. The base is used in an amount of about 1 to 20 times the molar amount of the compound []. The reaction is carried out at a temperature of 0 to 80°C, and usually completes in one hour to one week, although it varies depending on the type of base used, reaction temperature, etc. Step of compound [-1]→compound [-2] Compound [-2] is obtained by hydrolyzing compound [-1] in water and an inert solvent in the presence of a base. As the base, sodium hydrogen carbonate, ammonia, diethylamine, hydrazine, etc. are used. As the inert solvent, methanol, ethanol, tetrahydrofuran, dimethylformamide, ethyl acetate, acetone, chloroform, etc. are used. The base is used in an amount of 1 to 20 times the amount of compound [-1]. The reaction is carried out at 0 to 60°C and is usually completed in 1 to 10 hours. Process of compound [] → compound [-3] Compound [-3] is a compound [] and a compound (alkylating agent) represented by the general formula CH ₂ =CHZ (wherein, Z has the same meaning as above). It can be obtained by alkylating the 1a-position of the compound [] by reacting in an inert solvent. When using an active alkylating agent (e.g. acrolein), the reaction proceeds simply by mixing it with the compound [], but if necessary an organic base (e.g. triethylamine, sodium methoxide, potassium t-butoxide, etc.) may be used. ), the reaction proceeds quickly. Examples of solvents include ethers (ethyl ether, tetrahydrofuran, dioxane, ethylene glycol dimethyl ether, etc.), hydrocarbons (n-hexane, petroleum ether, benzene, etc.), amides (N/N-dimethylformamide, etc.), and esters (acetic acid). Organic solvents such as ethyl, etc.), ketones (acetone, etc.), halogenated hydrocarbons (methylene chloride, chloroform, etc.), dimethyl sulfoxide, etc. are used. The alkylating agent is used in a molar amount of 1 to 30 times the amount of compound []. The reaction is preferably carried out at 20 to 80°C, usually at 1
It will be completed within a week. Step of Compound [-3]→Compound [-4] The desired product can be obtained in the same manner as the step of Compound []→Compound []. Step of Compound [-4]→Compound [-5] The desired product can be obtained in the same manner as the step of Compound []→Compound [-1]. Process of compound [-5] → compound [-2] Compound [-2] can be obtained by subjecting compound [-5] to retroMichael reaction in the presence of an acid or base in a solvent inert to the reaction. . As the acid, an inorganic acid (for example, perchloric acid) is used, and as the base, a tertiary amine (for example, N.N-dimethylaniline) is used. Ethers (e.g. ethyl ether, tetrahydrofuran, ethylene glycol,
dimethyl ether), hydrocarbons (n-hexane, petroleum ether, benzene, etc.), amides (NN
-dimethylformamide, etc.), esters (ethyl acetate, etc.), ketones (acetone, etc.), halogenated hydrocarbons (methylene chloride, chloroform, etc.),
Dimethyl sulfoxide and the like are used. The acid or base is 1 to 1 for compound [-5]
Use about 100 times the molar amount. The reaction is preferably carried out at 0 to 80°C, usually at 10°C.
Finishes in minutes to several hours. Step of Compound [-4]→Compound [-6] The desired product can be obtained in the same manner as the step of Compound [-5]→Compound [-2]. Step of Compound [-6]→Compound [-2] The desired product can be obtained in the same manner as the step of Compound []→Compound [-1]. Step of compound [-2-1]→compound [-7] Compound [-7] is obtained by hydrolyzing compound [-2-1] in an aqueous solution of a base. As the base, inorganic bases (caustic soda, soda carbonate, etc.) and organic bases (triethylamine, etc.) are used. The concentration of base depends on the base used, but is usually
It is appropriate to use 0.01 to 1N. The reaction is usually carried out at room temperature and is completed in 30 minutes to several hours. Process of compound [-7] → compound [-2-2] Compound [-2-2] is a compound [-7] and diazoalkane (diazomethane, etc.), alkyl halide (methyl chloride, etc.), dialkyl sulfate (dimethyl sulfate, etc.) ) in a solvent (ethyl acetate, ethyl ether, etc.). When an acid is generated during the reaction, potassium carbonate, triethylamine, etc. are allowed to coexist as an acid binder. Process of compound [-2-2] → [-2-1] Compound [-2-1] is compound [-2-2]
and ammonia or an amine in an inert solvent (for example, methanol). Ammonia or amine is a compound [-2-
2] is used in a molar amount of 1 to 100 times. The reaction is usually carried out at 20 to 80°C and is completed in one to several hours. In each of the above steps, after the reaction is completed, the target product is recovered from the reaction solution by a conventional method, for example, by the method shown in the corresponding Examples and Reference Examples. Next, examples and reference examples will be shown. Example 1 (Compound []-[-1]) 1a-N-acetyl-10-decarbamoyl-10-
Dissolve 38 mg of methanesulfonyl mitomycin C in 8 ml of ethylene glycol dimethyl ether,
Add 150 mg of 1,5-diazabicyclo[5.4.0]undecene-5, and heat to reflux under a nitrogen stream for 2 hours. The reaction mixture is poured into saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The extract is washed with water, dried over anhydrous sodium sulfate, and the solvent is distilled off under reduced pressure. The residue was mixed with chloroform:acetone=6:
4 (volume ratio; the same applies to the following chromatography) is purified by silica gel column chromatography using a developing solvent. 24 mg of dark green crystals are obtained. The physical properties of this crystal are shown below. Mitomycin: gives molecular ion peak at m/e315. ¹ HNMR spectrum (in pyridine-d ₅ , δ
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 method, cm ^-1 ): 3435 (w),
3330 (m) (NH stretch.), 1695 (s),
1594 (vs), 1537 (vs) (C=O stretch.)
1656 (m) (C=C stretch.) is shown. According to the above, this substance is 1a-N-acetyl-
It is identified as 10-decarbamoyloxy-9-dehydromitomycin C. Yield 82%. Example 2 (Compound [-1] → [-2]) 57 mg of 1a-N-acetyl-10-decarbamoyloxy-9-dehydromitomycin C was dissolved in 1 ml of methanol, and 10% sodium hydrogen carbonate was dissolved in 1 ml of methanol.
ml and stirred at room temperature for 6 hours. After the reaction is completed, the reaction solution is extracted with ethyl acetate, dried over anhydrous sodium sulfate, and the solvent is distilled off under reduced pressure. The residue is purified by silica gel column chromatography using chloroform:acetone=1:1 as a developing solvent. 36 mg of dark green crystals are obtained. The physical properties of this crystal are shown below. Mass spectrum: gives molecular ion peak at m/e273. High-resolution mass spectrum results are actual measurements
2731130 (calculated value 2731113) ¹ HNMR spectrum (in pyridine-d ₅ , δ
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 method) (cm ^-1 ): 3420 (m)
(NH, stretch), 3320 (m) (NH,
stretch), 3285 (m) (NH, stretch), 1651
(m) (C=C, stretch), 1592 (vs) C=O,
stretch), 1532 (vs) (C=O, stretch) (Figure 1) From the above, this substance is 10-decarbamoyloxy-
It is identified as 9-dehydromitomycin C. Yield 73%. Example 3 (Compound [] → [-3]) 66 mg of 10-decarbamoyl mitomycin C was dissolved in 5 ml of methylene chloride, and acrolein (purity 90
%). Stir at room temperature for 4 days and remove the solvent under reduced pressure. The residue is purified by silica gel column chromatography using chloroform:methanol=93:7 as a developing solvent. black purple crystal
Get 76mg. The physical properties of this substance are shown below. ¹ HNMR spectrum (in pyridine-d ₅ , δ
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), 3.82 (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 method) (cm ^-1 ): 3430 (m)
(NH, stretch), 3335 (m) (NH,
stretch), 1722 (m) (C=O, stretch), 1604
(s) (C=O, stretch), 1554 (vs) (C=
O, stretch) From the above, this substance is 10-decarbamoyl-1a
-N-(2-formylethyl)mitomycin C
It is identified as Yield 97%. Example 4 (Compound [-3] → [-4]) 62 mg of 10-decarbamoyl-1a-(2-formylethyl) mitomycin C was dissolved in 0.33 g of anhydrous pyridine.
ml, add 15μ of methanesulfonic acid chloride under a nitrogen stream, and stir for 120 minutes while cooling with iced sodium monochloride. The reaction mixture is poured into saturated aqueous sodium bicarbonate solution and extracted with chloroform. After washing the extract with water, dry it with anhydrous sodium sulfate. After distilling off the solvent under reduced pressure, the residue is purified by silica gel column chromatography using chloroform:acetone=6:4 as a developing solvent. 44 mg of dark brown crystals are obtained. The physical properties of this substance are as follows. ¹ HNMR spectrum (in pyridine-d ₅ , δ
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) Infrared absorption spectrum (KBr tablet method) (cm ^-1 ): 3440
(m) (NH, stretch), 3340 (m) (N-
H, stretch), 1720 (m) (C=O, stretch),
1604 (vs) (C=O, stretch), 1555 (vs) (C
= O, stretch), 1351 (vs) (antisym, SO ₂ ,
strech), 1174 (vs) (sym., SO ₂ , stretch) From the above, this substance is 10-decarbamoyl-1a
It is identified as -N-(2-formylethyl)-10-methanesulfonyl mitomycin C. Yield 58%. Example 5 (Compound [-4]→[-5]) 10-decarbamoyl-1a-N-(2-formylethyl)-10-methanesulfonylmitomycin
Dissolve 52 mg of C in 10 ml of anhydrous ethylene glycol dimethyl ether, add 210 mg of 1,5-diazabicyclo[5,4,0]undecene-5, and heat under reflux for 2 hours under a nitrogen stream. The reaction mixture was poured into a saturated aqueous sodium bicarbonate solution, the extract was washed with ethyl acetate, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The residue is purified by silica gel column chromatography using chloroform:acetone=6:4 as a developing solvent. 16 mg of dark green crystals are obtained. The physical properties of this substance are shown below. ¹ HNMR spectrum (in pyridine-d ₅ , δ
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 method) (cm ^-1 ): 3420 (m)
(NH, stretch), 3225 (m) (NH,
stretch), 1718 (m) (C=O, stretch), 1649
(m) (C=C, stretch), 1535 (vs) (C=
O, stretch), 1535 (vs) (C=O, stretch) From the above, this substance is identified as 10-decarbamoyloxy-9-dehydro-1a-N-(2-formylethyl) mitomycin C. Yield 40%. Example 6 (Compound [-5]→[-2]) 10-decarbamoyloxy-9-dehydro-1a
-N-(2-formylethyl)mitomycin
Dissolve 26mg of C in 4ml of anhydrous methylene chloride,
- Add 100 mg of perchlorate of dimethylaniline;
Furthermore, add 0.4 ml of N.N-dimethylaniline. After stirring at room temperature for 15 minutes, the reaction mixture was poured into saturated aqueous sodium bicarbonate and extracted with chloroform. After washing the extract with water, it is dried over anhydrous sodium sulfate.
After distilling off the solvent under reduced pressure, the residue is purified by silica gel column chromatography using chloroform:acetone=1:1 as a developing solvent. 15 mg of dark green crystals are obtained. The mass spectrum, ¹ HNMR spectrum, and IR spectrum of this substance match those of the target compound of Example 2. Therefore, this substance is identified as 10-decarbamoyloxy-9-dehydromitomycin C. Yield 70%. Example 7 (Compound [-4]→[-6]) 10-decarbamoyl-1a-(2-formylethyl)-10-methanesulfonylmitomycin C33
Dissolve mg in 5 ml of anhydrous methylene chloride, and add 120 mg of perchlorate of N.N-dimethylaniline. Furthermore, 0.5 ml of N·N-dimethylaniline was added, and the mixture was stirred at room temperature for 30 minutes. The reaction solution was poured into a saturated aqueous sodium bicarbonate solution and extracted with chloroform. The extract is washed with water, dried over anhydrous sodium sulfate, and the solvent is distilled off under reduced pressure. The residue is purified by silica gel column chromatography using chloroform:methanol=97:3 as a developing solvent. Obtain 21 mg of reddish-purple crystals. The physical properties of this substance are shown below. ¹ HNMR spectrum (in pyridine-d ₅ , δ
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) and 7.57 (bs, 2H). IR spectrum (KBr tablet method) (cm ^-1 ) 3440 (w)
(NH, stretch), 3315 (m) (NH,
stretch), 1604 (vs) (C=0, stretch),
1555 (vs) (C=0, stretch), 1352 (vs)
(antisym., SO ₂ , stretch), 1174 (vs) (sym.
, SO ₂ , stretch) From the above, this substance is 10-decarbamoyl-10
-Identified as methanesulfonyl mitomycin C. Yield 72%. Example 8 (Compound [-6] → [-2]) 20.5 mg of 10-decarbamoyl-10-methanesulfonyl mitomycin C was added to anhydrous tetrahydrofuran.
Dissolve in 0.3 ml, add 30 mg of 1.5-diazabicyclo[5.4.0]undecene-5, and stir at room temperature for 40 hours. The reaction mixture was poured into saturated aqueous sodium bicarbonate and extracted with ethyl acetate. After washing the extract with water, it is dried over anhydrous sodium sulfate. After evaporating the solvent under reduced pressure, the residue was mixed with chloroform:acetone=1:1.
Purify by silica gel column chromatography using as a developing solvent. Dark green crystal 1.2mg
get. Mass spectrum of this substance, ¹ HNMR, IR,
TLC matches those of the product of Example 2. From the above, this substance is identified as 10-delcabamoyloxy-9-dehydromitomycin C. Yield 7.9%. Example 9 (Compound [-2-1] → [-7]) 18 mg of 10-dercarbamoyloxy-9-dehydromitomycin C was added to a 0.1N aqueous solution of caustic soda.
Dissolve in 3.75 ml and stir at room temperature for 45 minutes. After the reaction, adjust the pH to 4 with dilute hydrochloric acid and extract with ethyl acetate.
After washing the extract with water, dry it with anhydrous sodium sulfate,
The solvent is removed under reduced pressure. The residue is purified by silica gel column chromatography using chloroform:methanol=9:1 as a developing solvent. Obtain 12 mg of black-purple crystals. The physical properties of this substance are as follows. Ma spectrum: gives molecular ion peak at m/e274. ¹ HNMR spectrum (in pyridine-d ₅ , δ
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 method) (cm ^-1 ): 3285 (w)
(NH, stretch), 1644 (vs) (C=C,
stretch), 1630 (vs) (C=O, stretch),
1548 (vs) (C=0, stretch) (Figure 2) From the above, this substance is identified as 7-deamino-10-decarbamoyloxy-9-dehydro-7-hydroxy-mitomycin C. Yield 66%. Example 10 (Compound [-7] → [-2-2]) 7-deamino-10-decarbamoyloxy-9
-dehydro-7-hydroxymitomycin C12
Dissolve mg in 3 ml of ethyl acetate and cool with ice.
Add excess diazomethane-ether solution and let stand for 10 minutes. After removing the solvent under reduced pressure, chloroform:
Purify by silica gel chromatography using acetone = 6:4 as a developing solvent. black purple crystal 8
Get mg. The physical properties of this substance are as follows. Mass spectrum: gives molecular ion peak at m/e288. The high-resolution mass spectrum result is an actual measured value of 288.1120 (calculated value of 288.1110). ¹ HNMR spectrum (in pyridine-d ₅ , δ
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 method) (cm ^-1 ): 3305
(vw) (NH, stretch), 1650 (vs) (C=
0, stretch), 1554 (vs) (C=0, stretch)
(Figure 3) From the above, this substance is identified as 10-decarbamoyloxy-9-dehydromitomycin A. Yield 63%. Example 11 5.2 mg of 10-decarbamoyloxy-9-dehydromitomycin A is dissolved in 2 ml of saturated ammonia methanol solution and stirred at room temperature for 2 hours. After removing the solvent under reduced pressure, chloroform:acetone=1:1
Purify by silica gel chromatography using as a developing solvent. Obtain 4.1 mg of dark green crystals. Mass spectrum of this substance, ¹ HNMR, IR,
TLC was consistent with those of the target of Example 2.
From this, this substance is 10-decarbamoyloxy-
It is identified as 9-dehydromitomycin C. Yield 83%. Reference example 1 (Compound [] → []) Dissolve 230 mg of sodium in 50 ml of isopropanol, add 500 mg of mitomycin C, and dissolve at room temperature.
Stir for an hour. After neutralizing the reaction solution with excess dry ice and separating the precipitated sodium hydrogen carbonate, the solution is concentrated under reduced pressure. The residue is purified by silica gel column chromatography using chloroform:methanol=9:1 as a developing solvent. brown crystal
Get 270mg. This substance was identified as 10-decarbamoyl mitomycin C, a known substance, by its melting point, ¹ HNMR, TLC, etc. Yield 62%. Reference Example 2 (Compound [ ] → [ ]) Dissolve 138 mg of 10-decarbamoyl mitomycin C in 1 ml of pyridine, add 60 μl of acetic anhydride,
Cooled with ice-sodium chloride and under nitrogen stream 1
Stir for an hour. The reaction mixture is poured into saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. After washing the extract with water,
Dry over anhydrous sodium sulfate, and remove the solvent under reduced pressure. The residue is purified by silica gel column chromatography using chloroform:methanol=95:5 as a developing solvent. Obtain 153 mg of black-purple crystals. The physical properties of this substance are as follows. Mass spectrum: gives molecular ion peak at m/e333. ¹ HNMR spectrum (in pyridine-d ₅ , δ
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 method) (cm ^-1 ): 3435
(m), 3335 (m) (NH, stretch), 1694
(m), 1605 (s), 1555 (vs) (C=0,
stretch) From the above, this substance is identified as 1a-N-acetyl-10-dercarbamoylmitomycin C.
Yield 97%. Reference Example 3 (Compound [ ] → [ ]) Dissolve 62 mg of 1a-N-acetyl-10-decarbamoyl mitomycin C in 0.5 ml of anhydrous pyridine,
Add 15 μl of methanesulfonic acid chloride, and stir under a nitrogen stream while cooling with ice and sodium chloride. After 2 hours, the reaction mixture is poured into saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate.
After washing the extract with water, dry it with anhydrous sodium sulfate,
The solvent is removed under reduced pressure. The residue is purified by silica gel column chromatography using chloroform:methanol=96:4 as a developing solvent. Obtain 74 mg of black-purple crystals. The physical properties of this substance are as follows. ^1HNMR spectrum (in pyridine-d ₅ , δppm)
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 method) (cm ^-1 ): 3445
(w), 3235 (m) (NH, stretch), 1697
(s), 1605 (s), 1563 (vs) (C=0,
stretch), 1350 (vs) (antisym, SO ₂ ,
stretch), 1173 (vs) (smy.SO ₂ , stretch) According to the above, this substance is 1a-N-acetyl-
It is identified as 10-decarbamoyl-10-methanesulfonyl mitomycin C. Yield 97%.

[Brief explanation of the drawing]

FIG. 1 shows the infrared absorption spectrum of 10-decarbamoyloxy-9-dehydromitomycin C. FIG. 2 shows the infrared absorption spectrum of 7-deamino-10-decarbamoyloxy-9-dehydro-7-hydroxymitomycin C. Third
The figure shows the infrared absorption spectrum of 10-decarbamoyloxy-9-dehydromitomycin A.

Claims (1)

[Claims] 1. General formula [In the formula, X is amino, substituted amino, hydroxy or alkoxy, Y is hydrogen, -COR ₁ (in the formula, R ₁
represents hydrogen, alkyl, substituted alkyl, aryl or substituted aryl) or -CH ₂ CH ₂ Z (wherein Z represents alkyloxycarbonyl, acyl or cyano), R _A , R _B represents R _A is hydrogen; R _{B is -CH2OH or -CH2OSO2R2} (wherein _R2 represents alkyl, substituted alkyl or aryl)
Or, R _A and R _B together represent =CH ₂ . However, if R _A is H and R _B is -CH ₂ OH, Y is -
CH ₂ CH ₂ Z, and R _A is hydrogen and R _B is -
For CH ₂ OSO ₂ R ₂ , Y is hydrogen or −CH ₂ CH ₂ Z
It is. ] mitomycins. 2 General formula [Wherein, Xa is amino, substituted amino or alkoxy, _R2 is alkyl, substituted alkyl or aryl, and Y is hydrogen, _-COR1 (wherein, _R1 is hydrogen,
(represents alkyl, substituted alkyl, aryl or substituted aryl) or -CH ₂ CH ₂ Z (wherein Z represents alkyloxycarbonyl, acyl or cyano)] is dealkylated (or aryl) General formula characterized by sulfonation A method for producing a compound represented by the formula (wherein Xa and Y have the same meanings as above).