JPS61189283A - 7-n-aminomethylenemitomycin derivative, production thereof and antitumor agent - Google Patents

Abstract

NEW MATERIAL:The compound of formula I (R1 and R2 are H or lower alkyl; R3 is H and R4 is -CH2OCONH2 or group of formula II, or R3 and R4 together form =CH2; Y and Z are H or methyl; the wavy line represents alpha-bond or beta-bond; when R4 has beta-configuration, Y is H). EXAMPLE:9a-O-demethyl-7-N-dimethylaminomethylenemitomycin G. USE:An antitumor agent exhibiting antibacterial and antitumor activities. It has excellent effect especially to leukemia p-388. PREPARATION:The compound of formula I can be produced by the solvolysis of the compound of formula III at -30-+70 deg.C, using a catalyst selected from an inorganic salt such as alkali metal or alkaline earth metal carbonate or bicarbonate, and amine such as triethylamine, pyrrolidine, etc.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of the Invention The present invention relates to novel mitomycin derivatives having antibacterial and antitumor activities, methods for producing them, and antitumor agents containing them.

BACKGROUND OF THE INVENTION Mitomycins are generally known as antibiotics having antibacterial and antitumor activities. Typical mitomycins include mitomycin A1 mitomycin B1 mitomycin C1 porphyromycin (all of which are listed in the Merck Index 10th edition), mitomycin D1 and mitomycin E (all of which are listed in JP-A-1227-1227).
97. ), Mytoisin F (Unexamined Japanese Patent Publication No. 5
5-45322. ) etc. These mitomycins can be isolated from the culture fluid of Streptomyces kespitosus. Furthermore, 9-epi-mitomycin B and 9-epi-mitomycin D are chemically induced from mitomycin B (JP-A-56
-30978). Recently, the absolute configuration of mitomycin has been corrected (J, Am, Chem, Sac, 105
゜7199 (1983). Formulas A and B show the structures of the above various mitomycins modified accordingly (the same applies to 2C).

Formula Δ Main mitomycins derived from nature Mitomycin xA YA
ZA C-9A
0CR3CH3HβB OCH31
1CH3αCN1-12 CH3HβD
Nf12 It,
CH3αP NH2CH
3CH3αP OCH3CH3[:N
3 βJ OCH3CH,CH
3α volpiromycin NL C
H3CH3β2B 9β-mitomycin with hydroxyl group at position 9 9-epi-mitomycin B XB=O
CH39-epi-mitomycin D XB=NH
2 Furthermore, mitomycins with double bonds at the 9th and 10th positions are also known, and their production method is described in JP-A-55-15.
408.56-7787. Formula C shows the structure of double-bonded mitomycin.

Formula C double-bonded mitomycin mitomycin HXc”Ctls Yc=HZc=C
Hs mitomycin G XC-N)+2 YC=C
H3ZC=CH3 mitomycin K Xc:CN3Y
C=CH3Zc=CH39a-〇-demethylmitomycin G XC”N)+2 YC=HZc-C
Lla-demethylmitomycin G XC=N
H2Yo=CH3ZC-1 (1a-demethylmitomycin K XC-0CH3YC=CH3ZC=H
Although mitomycins have excellent antitumor activity, they also have side effects such as a decrease in white blood cells. Therefore, many derivatives have been synthesized with the aim of enhancing activity or reducing toxicity, and their biological properties have been investigated. I've been exposed to it.

Among these mitomycin derivatives, the amino group at position 7, which is relevant to the present invention, has been modified.
1).

J. Med. Chem., 26.16 (198
3), J. Med. Chem.

26.1453-(1983), J. Me'd, C.
An example is reported in Hem., 27.701 (1984). These documents also describe that mitomycin derivatives modified with an amino group at the 7-position exhibit antitumor activity in vivo. Among mitomycin derivatives modified with an amine group at the 7-position, an example of a compound closely related to the present invention is the recently published Japanese Patent Application Laid-open No. 5
9-1486. Among them, 7-(dimethylaminomethylene)amino-9a-methoxymitosan (hereinafter referred to as compound A) described in Example 8 of the patent is said to have particularly excellent antitumor activity. However, the compound described in the patent is a compound using mitomycin A1 mitomycin C1 porphyromycin and 7-N-methylmitomycin C as starting materials, and stereochemically, mitomycin C
It is limited to compounds having the same stereochemistry as (see JP-A-59-1486, page 10).

On the other hand, the mitomycin derivative having antitumor activity included in the present invention is represented by the following formula (I-1).

It is represented by (1-2) and (I-3).

(In the formula, R,, R2 represent a hydrogen atom or a lower alkyl group. Y and Z each represent a hydrogen atom or a methyl group, and V represents an α or β bond.

However, when the substituent at position 9 has a β configuration, Y represents a hydrogen atom.

Compound represented by formula (I-1) [hereinafter referred to as compound (I-1)
). Compounds with other large numbers are expressed in the same way. ] is synthesized using mitomycin B or an analog thereof as a raw material when the substituent at the 9-position is α. Such analogs include mitomycin D and mitomycin E, both of which have an amino group at the 7-position, but these can be obtained from nature as extremely small amounts of components during fermentation of mitomycin C (Japanese Patent Application Laid-open No. 122797-1989). ), is a compound that is difficult to obtain unless it is chemically derived from mitomycin B.

On the other hand, mitomycin B is a compound that is isolated as a trace component during mitomycin C fermentation, and it has been difficult to manually prepare an amount that can be used as a raw material for chemical synthesis. However, the research collaborators of the present inventors have investigated in detail the culture conditions and separation and purification methods for mitomycin C fermentation, with the aim of producing a larger amount of mitomycin B, and have significantly improved the fermentation unit of mitomycin B. succeeded in. Therefore, it has become possible to manually produce mitomycin B in large quantities at low cost, and the present invention, which can use mitomycin B as a starting material, has come to have important industrial significance.

Furthermore, when the substituent at the 9-position is β, it belongs to the generally well-known mitomycin. A representative example thereof is mitomycin C. However, in mitomycins in which the substituent at the 9-position is β, almost all mitomycins obtained from the natural world have a methoxy group at the 9a-position. Mitomycin (see formula A), which has the same stereochemistry as mitomycin C, i.e. 9-β and 9a-OH, is generally a difficult compound to handle, and the 9-epi-mitomycin D involved in the present invention is a special compound that can be obtained by using mitomycin B as a raw material.

As described above, the compound included in the present invention has an extremely unique chemical structure that is difficult to study even by those skilled in the art, even though it is called mitomycin. Mitomycin [Compound (I-3)]
The same applies to

Further, the compounds included in the present invention include the above-mentioned JP-A-59-1
It has been experimentally proven that it exhibits superior antitumor activity against leukemia p-388 compared to Compound Δ (shown in the reference example) described in No. 486 (described later). , there are clear characteristics compared to conventional techniques.

Problems to be Solved by the Present Invention As is clear from the documents cited above, a large number of mitomycin derivatives have already been synthesized, but there are still many mitomycin derivatives that have been synthesized. The development of excellent antitumor agents is desired. In the course of repeated research aimed at producing a substance with these properties, the present inventors also synthesized Compound A, independent of JP-A-59-1486, and noted that it had excellent antitumor activity. Was. And even more excellent antibacterial・E” Jf TO)-!-Il tongue skin・Rhinoceros Ai Kuroku:
As a result of repeated research aimed at producing nodomycin derivatives, a compound with biochemical properties superior to that of Compound A was discovered.
We were able to complete the present invention.

Means for Solving the Problems The present invention is based on the formula (T) (wherein R1 and R2 are the same or different and represent a hydrogen atom or a lower alkyl group. R3 and R4 together represent -CH2.Y and Z are the same or different and represent a hydrogen atom or a methyl group.

w represents an α bond or a β bond. However, when R1 has β coordination, Y represents a hydrogen atom. ) is related to a mitomycin derivative represented by

Compound (I> is composed of the aforementioned compounds (1-1), (1-2) and (I-3).

In formula (1), in the definition of R1 and R2, the lower alkyl group includes straight or branched pilates having 1 to 5 carbon atoms.

Next, a process formula for obtaining compound (Ii) is shown.

Process 1 Process 2 (I-2) -□-(II) Process 3
(IV) + (III) -1 (TI) Compound (1) is a mixture of compound (rJ) and compound (TI) in an inert solvent.
III). At this time, compounds (I-2) and (IV) are produced as by-products, but compound (I-2) can be led to compound (II) by selectively solvolyzing the 10-position 10CON=CHNR,R2. (Step 2). Compound (IV) is further compounded by (I
) under the same conditions as in step 1 to obtain compound (I-2), which can be led to compound (I-1) in step 3). The solvent used in step 1 is chloroform, dichloromethane, diethyl ether, tetrahydrofuran, dioxane, ethylene glycol dimethyl ether, acetonitrile, dimethyl sulfoxide, dimethylformamide, etc., and these may be used alone or in combination.

The reaction temperature and reaction time vary depending on the compound (IV) and the concentration of the reaction reagent, but are usually between -30°C and 7°C.
Several tens of minutes to several hours at a temperature of 0°C is sufficient.

As the solvent used in step 2, lower alcohols such as methanol, propatool, etc. are suitable.

Although these can be used alone, they can also be used in combination with ethers, acetonitrile, dimethyl sulfoxide, dimethylformamide, and the like. The reaction temperature and reaction time are usually in the range of -30°C to 70°C, and may be from several tens of minutes to several hours.

Although weak inorganic bases can be used as catalysts, amines that are weakly basic and highly sterically hindered are suitable.

Examples of the latter include aminodiphenylmethane and tertiary butylamine, but the solvolysis of such dialkylaminomethyleneimine groups is described in JP-A-5
9-1486.

Compound (I-1) is compound (■) (JP-A-56-778
7) can also be synthesized by the following process scheme using as a starting material.

That is, after reacting compound (V) with trichloroacetyl isocyanate to form compound (Vl) (step 4)
, Compound (1) is reacted in the same manner as in Step 1 to obtain Compound (■) (Step 5). Compound (■) is prepared by first reacting compound (V) and compound (III) to form compound (■).
It can be synthesized in the same manner by reacting trichloroacetyl isocyanate after that. Compound M) is obtained by solvolyzing the thus obtained compound (■) (Step 6). The trichloroacetylcarbamoylation in step 4 and the carbamoyl group production reaction in step 6 can be performed in the same manner as known methods (J, Natural Product 425
49 (197'9) and Proceedings of the 43rd Spring Annual Meeting of the Chemical Society of Japan, 910 pages, 1981, Tokyo). Solvents used in step 4 include chloroform, dichloromethane, diethyl ether, tetrahydrofuran, dioxane, ethylene glycol dimethyl ether, benzene, and toluene. These may be used alone or in combination. The reaction temperature and reaction time are usually in the range of -30°C to 30°C, and may be from several minutes to several hours. As the catalyst in step 6, inorganic salts such as carbonates or bicarbonates of alkali metals or alkaline earth metals, amines such as triethylamine, chloride (15), sopropylethylamine, pyrrolidine, piperidine, etc. are used. Reaction temperature, reaction Time is usually -3
The temperature may be in the range of 0°C to 70°C for several tens of minutes to several tens of hours. Step 5 is similar to step 1 above.

Next, a method for producing compound (1-3) will be explained.

Compound (I-3) is produced by reacting compound (IX) and compound (III) in an inert solvent. This step can be performed in the same manner as step 1 above.

Next, the present invention will be specifically explained with reference to Examples.

The physicochemical data of each compound was measured by the following instruments.

'H-NMR: JBOL FX-100) Lome-9-0M5: Old Tachi M-80B mass spectrometer.

IR: Shimazu IR-27-G and JA
SCOIR810゜Silica gel TLC: Mark
Art 5714゜Example 19a-0-demethyl-7
-N-dimethylaminomethylene mitomycin G (compound 1) To 1 ml of dried dimethylformamide (hereinafter abbreviated as DMF), 0.1 ml of dimethylformamide dimethyl acetal (hereinafter abbreviated as DMFA) and 090 mg of 9a-〇-demethylmitomycin were added, and nitrogen Stir for 6 hours at room temperature under air flow.

After distilling off the solvent under reduced pressure, C)(C1,-MeOH(
The product is separated by silica gel column chromatography using 98:2 v/v) as a developing solvent.

The product is purified again by silica gel column chromatography, and the solution eluted with AcOEt-MeOH (99:IV/V) is concentrated under reduced pressure to isolate a dark green paste. When this material is dissolved in a small amount of CHcL and the solution is added dropwise into cyclohexane, Compound 1 precipitates out as a green precipitate.

This was separated in a furnace to obtain 81 mg of Compound 1. Yield 75%
.

'H-NMR (CH2DCONHCJ!3): δ1.8
6 (3N, s).

2.19 (3H, S), 2.26 (2H, m), 3
．． 03 (311, S), 3.07 (3tl, s),
3.48 (lft, dd, J=12.7.1.3)
.

4.17 (LH, d, J=12.7), 5.44 (
IH, d, J=0.5).

I R (KB r ): 3400. 292B,
2854. 1644. 1620°1529, 1
376, 1306. 1113. 1054cm-'
.

MS (m/z): 328 (M”) 311
．． 284. 295° high resolution MS: M” =
328.1517 (calculated for CI7H2ON403 -328,1533).

TLC (CHCA3 MeOH, 19: 1 v
/v): Rf=0.40° Example 2 53 mg of 1a-demethyl-7-N-dimethylaminomethylene mitomycin G (compound 2)
Example 1 using demethylmitomycin G as a starting material
Compound 247m was obtained as a green powder by almost the same procedure as
g is obtained.

Yield 74%.

'H-NMR(CH2DCONH C13)
: δ1.95 (3H, s).

2.85 (28, br, s), 3.05 (3t (
, s), 3. IQ (3ft, s).

3.11 (3H, s), 3.55 (IH, br, d,
J=12.9).

4.27 (1N, d, J=12.9), 5.42
(11(,d, J=0.6).

6.22 (IH, d, J=0.6), 7.74 (I
H, 5) IR (KBr): 3292.2930.2
854.1645.1622゜1599, 1572.1
532.1435.1376, 1304.1254゜1
214, 1111.1083.1053.939 cm
-'.

MS (m/z): 328 (M+), 313.2
97.282,254° High resolution MS: M”=
328.1532 (c, 7■2oN, 111 matches against male balls on o3 = 3981 et al. 34) -TLC (CHCβ3
MeOH, 19: 1 v/v): Rf=
0.48° Example 3 7-N-dimethylaminomethylene mitomycin D (compound 3a) and 7-N,Nl0-his(dimethylaminomethylene) mitomycin D (compound 3b) (In the naming of the present compounds, N10 is mitomycin This refers to the nitrogen atom of the carbamoyloxy group at position 10. The same applies below.) D, dry DMFO containing 80 μp of MFA, 4 m
83 mg of mitomycin D was dissolved in 1 liter of water and stirred for 40 minutes at room temperature under a nitrogen stream.

After distilling off the solvent under reduced pressure, it was subjected to silica gel column chromatography, and CHCβ3 MeOH (95:5V
Collect the green fraction (referred to as fraction A) eluting at /v). Continuing, CHCj? 3MeO)1(93:7
v/v) and collect another green fraction (referred to as fraction B). Finally, remove the solvent from CHC13-CH30H (85
: 15 v/v) and collect the resulting fractions, from which 26 mg of mitomycin D is recovered.

After fraction B is concentrated, it is purified again by silica gel column chromatography. ACOEt-M eOH (9
The green fractions eluting at 2:8 v/v) are collected and concentrated to yield 25 mg of compound 3a as a dark green powder. Yield 45%.

'H-NMR (CH2DCONHCA3): δ1.9
0 (3H, s).

2.25 (511,s), 3.03 (311,s),
3.08 (311, s).

3, 46 (LH, br, d, J=12.7), 3
．． 7H111,t, J=4.2)4.08(lft,
d, J=12.7), 4.20 (IH, br,)
.

4.70 (21i, d, J=4.2),
4.72 (2t i, br, s).

7.70 (LH, s).

I R (KB r ) +3430.2930.173
0.1621.1535°1310cm-'.

MS (m/z) +389 (M”), 371.
346.328° 310, 295.273.259° High resolution MS: M”-389,1694 (C,,
Calculated value for H23N505 -389,1697).

TLC (CHCII3 MeOH, 19: 1 y/v
): Rf=0.17° After fraction A is concentrated, it is further purified by preparative silica gel thin layer chromatography. AcOEt-MeOl
Expand with l (] 9 : 1 v/v), Rf=0
．． The green band of 09 is collected to give 26 mg of compound 3b as a dark green powder. Yield 34%.

'H-NMR(CH2DCONH(13):61.90
(311, S).

2, 25 (311s), 2.28 (2H, m), 3
．． 03 (3H, s).

3.05 (3H,s), 3.07 (3H,s),
3.1H311,s).

3.48 (LH, dd, J=12.7. 1.7)
, 3.78 (ill, m).

4.08(], H, d, J=12.7), 4
．． 51 (1N, br,).

4.75(2)1. m>, 7.70(II(
, s), 8.38 (IN, s>.

I R (KB r ) +3380. 2920.
1617. 1526. 1419.

1374, 1309. 1263. 1226,11
13. 1076, 1059 cm'.

MS (m/, Z): 444 (M”,
Calculated value for C21828NBO5 -444), 4
26.328.

TLC (CHCj!3 MeOH, 19: 1 v
/v): Rf=0.29゜li Example 4 7-N-dimethylaminomethylene mitomycin E (compound 4a) and 7-N,N''-bis(dimethylaminomethylene) mitomycin E (compound 4b)
.

Dissolve 39 mg of mitomycin E in 4 ml of dry HD U F O containing 80# DMFA and stir for 1 hour and 20 minutes at room temperature under nitrogen flow. After distilling off the solvent under reduced pressure, column chromatography is performed using CH (13-MeOH (97: 3 v/v) as an eluent) to collect a green fraction (referred to as fraction A). , continued C)l(J!
Elution with 3-MeOH (92:8 v/v) gives another blue fraction (designated fraction B).

Fraction B was concentrated and purified by silica gel thin layer chromatography to recover 15 mg of mitomycin E. After concentration, fraction A was subjected to silica gel thin layer chromatography and developed with ΔcOEt-MeOH (19:IV/V). Compound 4a dark green powder (25% yield) as 7 mg dark green powder from green band at Rf=0.16
and 4 mg of dark green powder (yield 13%) of compound 4b are isolated from the green hand with Rf=0.02, respectively.

Compound 4a'H-NMR (CH2DCONH (J!3): δ1.
91 (31-1, s).

2.20 (LH, d, J=4.6), 2.31 (
3H,s), 2.36(iH.

dd, J=4.6.2.0), 3.03(3H,S
), 3.07 (3H,S).

3.31 (3tl, s), 3.56 (ltl, d
d, J=12.7.2.0).

3.84 (III, dd, J=9.5.3.9),
3.95 (LH, d, J=12.7).

4.43 (11-1, dd, J=10.7.9.5)
, 4.6N2)1. br, s).

4.84 (LH, dd, J=10.7.3.9),
7.68 (ill, s).

IR(KBr) :3450.3362.2922
．． 1713.1625°1548, 1538.1326
．． 1304.1117.1074.1051 cm-'.

MS (m/z): 403 (M”), 372.
342.327° 311, 295° High resolution MS: M+= 403.1871 (C,
Calculated value for 91125N505 -403,1854
>.

Compound 4b'H-NMR (CH2DCONHCβ3); δ1.9H
311, s).

2.3H3H,s), 2.33(211,m),
3.02 (3ft, s).

That is, after reacting compound (V) with trichloroacetyl isocyanate to form compound (Vl) (step 4)
, Compound (I) is reacted in the same manner as in Step 1 to obtain Compound (■) (Step 5). Compound (■) can be synthesized in the same manner by first reacting compound (V) and compound (I) to form compound (■) and then reacting it with trichloroacetyl isocyanate. By solvolyzing the compound (■) thus obtained, the compound (
1) is obtained (Step 6). The trichloroacetylcarbamoylation in step 4 and the carbamoyl group production reaction in step 6 can be performed in the same manner as known methods (J, Natural Product 42549
(197'9) and the 43rd Spring Annual Meeting of the Chemical Society of Japan,
Proceedings 910 pages, 1981, Tokyo). Solvents used in step 4 include chloroform, dichloromethane, dietheretherepetetrahydrofuran, dioxane, ethylene glycol dimethyl ether, benzene, and toluene. These may be used alone or in combination. The reaction temperature and reaction time are usually in the range of -30°C to 30°C, and may be from several minutes to several hours. As the catalyst in step 6, inorganic salts such as carbonates or bicarbonates of alkali metals or alkaline earth metals, amines such as triethylamine, diisopropylethylamine, pyrrolidine, piperidine, etc. are used. Reaction temperature and reaction time are usually -30℃ to 70℃
It may take from several tens of minutes to several tens of hours. Step 5 is similar to step 1 above.

Next, a method for producing compound (13) will be explained.

Compound (I-3) is produced by reacting compound (IX) and compound (I) in an inert solvent. This step can be performed in the same manner as step 1 above.

Next, the present invention will be specifically explained with reference to Examples.

The physicochemical data of each compound was measured by the following instruments.

'H-NMR: JBDL PX-100 Shek) 0 meters.

MS: Hitachi M-808 mass spectrometer.

IR: Shimazu IR-27-G and J SCH2DCONH IR810゜1J Silica Gel TLC: Merk Art 5714
Example 19a - 0-Demethyl-7-N-dimethylaminomethylene mitomycin G (compound 1) Add 0.1 ml of dimethylformamide dimethylacecool (hereinafter abbreviated as DMFA) to 1 ml of dried dimethylformamide (hereinafter abbreviated as DMF). 9a-〇-demethylmitomycin G
90. mg and stirred at room temperature for 6 hours under nitrogen flow.

After distilling off the solvent under reduced pressure, CHCRs M e OH
The product is separated by silica gel column chromatography using (98:2 v/v) as a developing solvent.

The product is purified again by silica gel column chromatography, and the solution eluted with AcOEt-MeOH (99:IV/V) is concentrated under reduced pressure to isolate a dark green paste. When this material is dissolved in a small amount of CHCl3 and the solution is added dropwise into cyclohexane, compound 1 precipitates out as a green precipitate.

This was sent door to door to obtain 81 mg of Compound 1. Yield 75%
.

'H-NMR(CH2DCONH CI 3)
: δ1.86 (3H, S).

2.19 (3H, s), 2.26 (28,'m),
3.03 (3H, s).

3.07 (3H, s), 3.48 (ltl, dd,
J=12.7.1.3).

IR (KBr) +3400. 2928. 285
4. 1644. 1620°1529, 1376,
1306. 1113. 1054cm-'.

MS (m/z): 328 (M+) 311. 2
84. 295° high resolution MS: M”-328,
1517 (c, calculated value for LoLO3 -328,1
533).

TLC (CHCj2s M eOH, 19: 1
v/v ): Rf = 0.40° Example 2 53 mg of 1a-demethyl-7-N-dimethylaminomethylene mitomycin G (compound 2)
Example 1 using demethylmitomycin G as a starting material
Compound 247m was obtained as a green powder by almost the same procedure as
g is obtained.

Yield 74%.

'H-NMR (CH2DCONHCl2): δ1.9
5 (3N, s).

2.85 (2H, br, s), 3.05 (3H, s)
, 3.10 (3fl, s).

3.1H3H,s), 3.55(IH,br,d,
J=12.9).

4.27 (18, d, J=12.9), 5.42 (
IH, d, J=0.6).

6.22 (IH, d, J=0.6), 7.74 (I
H, S) I R (KB r ) +3292.293
0.2854.1645.1622゜1599, 157
2.1532.1435.1376, 1304.125
4゜1214, 1111.1083.1053.939
cm-'.

MS (m/z): 328 (Ma, 313.29
7.282,254°calculated value -389), 371.3
46.328.295°TLC (CHCI3-CH30
H,9: 1 v/v): Rf=0.33° Fraction A is concentrated to yield 40.3 mg of compound 6d as a dark green solid. Yield 36%.

It is clear from the following fact that the compound obtained from fraction A is 6d.

40.3 mg of the solid obtained from Δ was added to 1 ml of CH,O
Dissolved in H and added 4 mg K 2 CO 3 at room temperature.
Stir for 0 minutes. Add 9 ml of CHCA3 to the reaction solution,
Chromatography is performed using a short silica gel column, eluting with CHC13-MeOH (9:1 v/v). The eluate was concentrated under reduced pressure and again CHCl3-'CH3
Silica gel column chromatography is performed using 0H (92:8 v/v), and the eluted green band portion is concentrated under reduced pressure. A small amount of n-hexane is added to the residue, and the solvent is distilled off again under reduced pressure and dried to obtain 13.8 mg of dark green powder. The physicochemical properties of this substance are those of the above compound 6C.
is the same as Yield 47%.

Example 7 The antibacterial activity of some compounds included in the present invention against various bacteria is shown in Table 1 in terms of minimum growth inhibitory concentration (μg/ml). The minimum growth inhibitory concentration is determined by the agar dilution method.
Measured at H7.0. The symbols used for bacteria in the table are as follows.

SF Streptococcus faecalis ATCC10
541, sA Sphphylococcus aureus ΔTC
C6538P, BS Bacillus subtilis 107
07. PV Proteus vulgaris ATCC689
7, SS Shigella Sonnei ATCC924) O, K
P Klebscilla pneumoniae ATCC10031
MM-C in the table represents mitomycin C.

Table 1 Antibacterial activity (minimum inhibitory concentration, x/ml>Example 8 Antitumor activity and toxicity against Sarcoma 180 solid tumor.

Table 2 shows the antitumor activity (EDso), acute toxicity (LDso), and effect on peripheral white blood cell count (WBC4000) against Sarcoma 180 solid tumor using some compounds included in the present invention as examples. Ta.

WBC,. . . represents the dose of drug that reduces the peripheral leukocyte count to 4000/mm3. All experiments were conducted using ddy mice, and specific experimental procedures are shown in Japanese Patent Application No. 12428.

Table 2 Example 9 Antitumor activity against leukemia P-388 P-388 ascites tumor bile cancer mouse (DBA/2) 7 days after transplantation
Ascitic fluid was collected from the peritoneal cavity of the patient. The number of P-388 cells in this ascites was counted, and a suspension of 5 x 106 cells/ml of tumor cells was prepared using sterile physiological saline, and 0.2 ml of it (
I x 106 cells), weighing 20-25
g CH2DCONHF was implanted intraperitoneally into mice.

CH2DCONHF of 6 animals per group 24 hours after tumor implantation
The test drug was intraperitoneally administered once to mice, and the number of survival days was observed for 33 days. The effectiveness of the drug was determined by the ratio of the average survival days on the 333rd day after transplantation to the average survival days of the control group (untreated group) (survival prolongation rate, ILS%, Increased
Life 5pan) and the results are shown in Table 3. Table 4 also shows the results of a similar experiment conducted using P-388, which is resistant to mitomycin C. From Tables 3 and 4, the maximum life extension rate of Compound 3a is 213%, respectively.

143%, compared with 75% and 73% of Compound A (synthesis example shown in Reference Example), it is clear that 3a has superior antitumor activity. In comparison, superior clinical effects can be expected. In addition, in a similar experiment, the ILS of Compound 1 at a dose of 10 mg/kg was 83%.

Table 3 Effect on P-383leukemia (ILS%) Dose Compound 3a Compound Δmg/kg 0, 0625 150, 125
480, 25
2fl 450, 5 53
631, 0 58
682, 0 58 754.0
145 206, 0
213 - Table 4 Mitomycin C resistance p
-Effect on 3381eukelTlia (II, 8
%) Reference example? -N-dimethylaminomethylene mitomycin C (compound A; compound described in Example 8 of JP-A-59-1486) Compound Δ is obtained by the same operation as in Example 3 using mitomycin C as a raw material.

'H-NMR (CH2DCONH(13): δ1.93
(3H,s).

2.80 (IH, dd, J=4.4.2.0), 2.
90 (IH, d, J=4.4).

3.04 (3H, s), 3.08 (38, s), 3
．． 22 (3H, S).

3.50 (IH, dd, J=12, ?, 2.0),
3.61 (IH, dd.

J=io, 5.4.4), 4.20(LH, d, J
= 12. n.

4.50 (IH, dd; δ40.7.10.5>,
4.74 (2N, br,).

4.78 (11 (, aa, J=10.7.4.4),
7.69 (ltl, s).

I R (KB r ): 3310.2940.17
18.1624.15401306, 1059cm-'
.

MS (m/z): 389 (M”), 357
．． 346. 328°313, 297.

High resolution MS: M+=389.1720 (c,
Calculated value for art□3NSO5 = 389.1697
).

TLC (CHCR3M e OH19: lv/v)
:Rf=0.40° Effect of the invention Compound (1) has excellent antibacterial and antitumor effects.

Patent applicant (102) Representative of Kyowa Hakko Kogyo Co., Ltd.
Mikio Kato 32) Procedural amendment 1, Indication of case 1985 Patent Application No. 31074 2, Title of invention 7-N-aminomethylene mitomycin derivative, manufacturing method and antitumor agent 3, Person making amendment case Relationship with Patent applicant postal code 100 Address 6-1 Otemachi-chome, Chiyoda-ku, Tokyo Name (
102>Kyowa Hakko Kogyo Co., Ltd.

/nI Q nguest Q4:, trz r/ ll:oZ
l? Digitize the Q OZI I-.

(3) Correct rl 730J on page 20, line 9 to rl 703.

(4) r3 on page 23, line 1. o 2 (3H, s), delete J.

Procedural amendment 1985? Month/Date 1, Indication of the case 1985 Patent Application No. 31074 2, Name of the invention 7-N-aminomethylene mitomycin derivative, manufacturing method and antitumor agent 3, Person making the amendment Relationship with the case Patent applicant's mail Number 100 Address 6-1 Otemachi-chome, Chiyoda-ku, Tokyo Name (
102) Column 5 of the detailed description of the invention in the Kyowa Hakko Kogyo Co., Ltd. specification, contents of amendment (1) "1-propyl" on page 10, line T1 to page 11, line 1 (3) r on page 12, line 3 (I)J r(TI)J
Correct.

(4) r (rV) J in the 7th line directly below 12 as r(II
), corrected to J.

(5) Change “rMerk” on page 17, line 1 to “MerCk”
Correct to.

Claims (8)

[Claims] (1) Formula (I) ▲There are mathematical formulas, chemical formulas, tables, etc.▼(I) (In the formula, R_1 and R_2 are the same or different and represent a hydrogen atom or a lower alkyl group. R_3 and R_4 are R_
3 is a hydrogen atom and R_4 represents -CH_2OCONH_2 or ▲There are mathematical formulas, chemical formulas, tables, etc.▼, or R_3 and R_4 together represent =CH_2. Y and Z are the same or different and represent a hydrogen atom or a methyl group. ■ represents an α-bond or a β-bond. However, when R_4 has β coordination, Y represents a hydrogen atom. ) mitomycin derivative. (2) In formula (I), R_3 is a hydrogen atom and R_4 is -CH
The mitomycin derivative according to claim 1, which is _2DCONH_2 [hereinafter referred to as compound (I-1)]. (3) R_3 is β-hydrogen atom and R_4 is α-CH_2O
The mitomycin derivative according to claim 2, which is CONH_2. (4) The mitomycin derivative according to claim 2, wherein R_1, R_2 and Z are methyl groups, and Y is a hydrogen atom. (5) The mitomycin derivative according to claim 1, wherein in formula (I), R_3 is a hydrogen atom and R_4 is ▲a numerical formula, a chemical formula, a table, etc.▼. (6) The mitomycin derivative according to claim 1, wherein R_3 and R_4 together represent =CH_2 in formula (I). (7) A compound characterized by solvolyzing a compound represented by the formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (In the formula, R_1, R_2, Y, Z and ■ have the same meanings as above) (I-1) Production method. (8) An antitumor agent containing a mitomycin derivative represented by formula (I) as an active ingredient.