JPH0472838B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to mitomycin analogues having one or more amidino groups. These compounds are mitomycin C derivatives in which either the 7-amino group and the carbamide nitrogen atom, or both, are part of the amidino substituent. These compounds are active antitumor agents in experimental animal tumors. Nomenclature: Systematic Chemical Abstracts for Mitomycin C
The name is: [1a R -(1aα, 8β, 8aα, 8bα)]-6-amino-8-[((aminocarbonyl)oxy)methyl]-1,1a,2,8,8a,8b-hexahydro-8a -methoxy-5-methyl-azilino [2',
3',3,4]-pyrrolo[1,2-a]indole-4,7-dione. Accordingly, the azilinopyrroloindole ring system is numbered as follows: The conventional nomenclature system that has been widely used in the literature for mitomycin treats the aforementioned ring system containing mitomycin's characteristic substituents as mitosane. Mitosane This system is convenient and suitable for a number of simple derivatives, such as those with N-substituents on the azilino ring nitrogen atom or in the 7- or 9a-position, but for general applications. It has certain ambiguity and shortcomings. Regarding the compounds of the present invention,
Some have substituents on both the azilino ring nitrogen atom and the side chain carbamoyl nitrogen atom, and there is no convenient numbering method to distinguish these positions. Therefore, we have chosen herein to use the mitozane nomenclature system and refer to the azilino nitrogen atom as N ^1a and the carbamoyl nitrogen atom as N ¹⁰ . With regard to the stereochemical configuration of the products of the invention, when identified by the basic name "Mitozan" or by the structural formula, it is assumed that the stereochemical configuration is the same as that of mitomycin C. . Mitomycin C is produced by fermentation and is already licensed in the United States for the treatment of widespread adenocarcinoma of the stomach and pancreas in proven combinations with other approved chemotherapeutic agents, as well as other similar agents. It is currently marketed for temporary relief when it is ineffective.
(Mutamycin Pristle Laboratories, Shiraki Youth, New York
13201, Physician's Death Cliff Arrangement 35th Edition 1981, pages 717 and 718 [Bristol
Laboratories, Syracuse, New York 13201,
Physicians′ Desk Reference〕）Might Machine C
and its production by fermentation are the subject matter of US Pat. The structures of mitomycin A, B, C and porfiromycin were obtained from American Cyanamid's Lederle Laboratories Division.
J.Amer.Chem.Soc. 84 , 3185-3187 (1962)
was first published in. One of the chemical transformations used in this structural study for mitomycin A and mitomycin C was by reaction of the former, 7-9α-dimethoxymitozane, with ammonia, to form the latter, 7-amino-9α-methoxymitozane. It was a conversion to Substitution of the 7-methoxy group of mitomycin A has proved to be a rather interesting reaction in the preparation of antitumor active derivatives of mitomycin C. The following documents and patents each deal with the conversion of mitomycin A into 7-substituted aminomitomacin C derivatives with antitumor activity. The aim of this work was to produce derivatives that are more active and, in particular, less toxic than mitomycin C. Matsui et al., “The Journal of Antibiotics”, XI, 189-198 (1968). Kinoshita et al., “J.Med.Chem.” 14 , 103−109
(1971). Iyengar et al., “J.Med.Chem.” 24 , 975−981
(1981). Iyengar, Sami, Remers, et al., “Bradner,
Abstracts of Papers Annual Meeting ef the
American Chemical Society, Las Vegas,
Nevada, March 1982, Abstract No.MEDI72, the following patent deals with the preparation of 7-substituted aminomitozans by the reaction of mitomycin A, mitomycin B or their N ^1a -substituted derivatives with primary or secondary amines. There is. Cosulich et al.: U.S. Patent No. 3,332,944 (July 1967)
Matsui et al.: U.S. Patent No. 3420846 (January 7, 1969) Matsui et al.: U.S. Patent No. 3450705 (June 1969)
17th) Matsui et al.: U.S. Patent No. 3514452 (May 1970)
26th) Nakano et al.: U.S. Patent No. 4,231,936 (November 1980)
(May 4, 1981) Remers: U.S. Patent No. 4,268,676 (May 1981)
(19th) Mitomycin C derivatives with an amino substituent substituted at position 7 have also been produced by direct biosynthesis. The method is to supplement the fermented broth with a range of primary amines and carry out a conventional mitomycin fermentation. (CAClaridge et al.
Abst.of the Annual Meeting of Amer.Soc.for
Microbiology1982.Abs.028) Mitomycin C is the primary mitomycin produced by fermentation and is also a commercially available form. The current technology for the conversion of mitomycin C to mitomycin A, which is used for the preparation of the semi-synthetic substituted amino analogues of mitomycin C mentioned in the aforementioned publications, is to convert mitomycin C into the corresponding 7-hydroxymitosan (extremely This process involves hydrolyzing the substance into an unstable substance (an unstable substance) and then methylating this substance with diazomethane, an extremely dangerous substance to handle. One attempt to avoid the use of diazomethane in the methylation of 7-O-demethylmitomycin A obtained by hydrolysis of mitomycin C involves the use of 7-acyloxymitozanes (Kyowa Hakko Kogyo KK: Japanese Patent Publication) , No. 56-73085, Farmdoc No. 56227D/31). The present invention provides monomers of mitomycin C in which either or both the 7-amino nitrogen atom and the N ¹⁰ carbamoyl nitrogen atom are part of an amidino substituent, or the 7-amino nitrogen is part of a guanidino group. A novel family of guanidino or mono- and bis-amidino analogs. Corresponding analogues of mitomycin A having a methoxy group in the 7 position and an amidino group in the N ¹⁰ position are also encompassed by the present invention. The compounds of the present invention are represented by the following structural formula (). [However: A is amino, methoxy, hydroxy,
(1-lower alkyl-2(1H)-pyridinylidene)
Amino, or a group represented by the formula [Formula] [Formula] [Formula] [Formula] [Formula] [Formula] [Formula], B is amino or an amidino group of the formula [Formula], and A or B at least one of the groups specified above other than amino, methoxy, and hydroxy, n is an integer of 0, 1, 2, or 3, and R ¹ is hydrogen, lower alkyl, or lower alkanoyl; , R ² is hydrogen or lower alkyl, R ³ is lower alkyl, lower alkoxy, or R ⁴ and, together with the nitrogen atom to which they are bonded, pyrrolidine, 2-, or 3-lower alkylpyrrolidine, piperidine, 2-, 3- or 4-lower alkylpiperidine, 2,6-di-lower alkylpiperidine, piperazine, 4-substituted piperazine (provided that the 4-substituent is an alkyl or carbalkoxy group each having 1 to 8 carbon atoms) ; phenyl, methylphenyl, methoxyphenyl, halofenyl,
nitrophenyl or benzyl), azepine, 2-, 3-, 4-, or 5-lower alkyl azepine, morpholine, thiomorpholine, thiomorpholine-1-oxide or thiomorpholine-1,1-dioxide. ^pyrrolidine , ²
- or 3-lower alkylpyrrolidine, piperidine, 2-, 3-, or 4-lower alkylpiperidine, 2,6-dilower alkylpiperidine, piperazine, 4-substituted piperazine (however, each of the 4-substituents is Alkyl or carbalkoxy with 8 carbon atoms; phenyl, methylphenyl, methoxyphenyl, halofenyl, nitrophenyl,
or benzyl), azepine, 2-, 3-,
4- or 5-lower alkyl azepine, morpholine, thiomorpholine, thiomorpholine-1-oxide, or thiomorpholine-1,1-dioxide is formed, and R ⁵ is C _1-18 alkyl other than tertiary alkyl; and R ⁷ and R ⁹ are independently H or lower alkyl, provided that the aforementioned lower alkyl, lower alkanoyl and lower alkoxy groups have 1 to 6 carbon atoms]. Those skilled in the art will recognize that tautomers exist for some of the aforementioned amidino groups. Such materials are also intended to be included in the foregoing formulas and the present invention. The aforementioned substances of formula () have antitumor activity in experimental animals. These substances are also useful as intermediates in the production of other compounds that have antitumor activity in animals. The present invention includes methods for producing the aforementioned substances and methods for converting the aforementioned substances into other useful compounds having antitumor activity in experimental animals, as described below. The method of the invention using the aforementioned substances as intermediates for the production of other compounds having antitumor activity in animals comprises compounds of formula () in which A or both A and B are said amidino groups. is reacted with a primary amine, resulting in cleavage of the N ¹⁰ -aminomethylene substituent, if present, resulting in mitomycin A
and its conversion into NH ₂ groups such as those present in mitomycin C. With some exceptions,
The primary amine also reacts by displacement of the amidino group at position 7, replacing it with an amino substituent corresponding to the reactant. These processes are expressed by the following equation. R ⁵ of the primary amine, which represents the nitrogen substituent of various known and novel 7-substituted mitomycin C compounds described herein, and which can convert the 7-amidino group of formula (), may be any group other than tertiary alkyl. C _1-18 alkyl. N ¹⁰ Represents a nitrogenous substituent of a primary amine in which only cleavage of the amidino substituent is possible
R ⁶ is the remainder of a very weakly basic aliphatic amine or a highly hindered alkyl or aralkyl amine. Examples are trifluoroethylamine, benzhydrylamine (ie aminodiphenylmethane) or tertiary-butyl-amine. Compounds of formula () are prepared by reaction of mitomycin C, 7-hydroxy-9a-methoxymitozane, or mitomycin A or a N ^1a -substituted analogue of any of the above substances with an amide acetal. A compound of formula () in which A instead of B is the amidino group is mitomycin C or its
The N ^1a substituent is reacted with a strong base to form an anion at N ⁷ , and then the anion is reacted with a reagent capable of producing an aminoethylene group, such as a halomethyleniminium salt. It can be manufactured even if Preferred compounds of the invention are mitomycin C analogues in which a 7-amino group is incorporated into a substituted or unsubstituted amidino group. These compounds have strong antitumor effects against tumors in experimental animals.
These compounds are prepared by reacting mitomycin C with a reagent capable of converting a 7-amino group to a 7-amidino group. Preferred reagents for this purpose are:
Mitomycin C in good yield under mild conditions
(Example 1)
-5, 18). Other groups of amidine-forming reagents include imidoyl halides (Example 17), halomethyleneiminium salts (Ex. Example 15), 2-halo-1-alkylpyridinium halide (Example 16) and iminoether or iminothioether (Example 13)
and 14). The conditions for deelectrification of mitomycin C are to treat a solution of mitomycin C in dimethylformamide with about 1.5 molar proportions of sodium hydride at room temperature. The reaction of the anionic form thus formed with one of the aforementioned reagents is
Preferably 1 to 1.5 for mitomycin C
It is carried out using molar proportions of reagents and at temperatures from room temperature to about -60°C. A neutral polar organic solvent such as dimethylformamide, hexamethylphosphoamide, dimethylsulfoxide or pyridine is used as the reaction medium. However, the method should not be limited to the formation of anionic mitomycin C in this particular manner. This is because improvements are easy for those skilled in the art. A preferred method for producing a compound of formula () in which B or each of A and B is ^an amidino group of formula (However, R ² , R ³ and R ⁴ are as above,
R ⁸ is lower alkyl or cycloalkyl having up to 6 carbon atoms, or two R ⁸ groups are joined to form an alkylene chain, with two oxygen atoms and an intervening carbon atom forming a 5- to 6-membered (forming a cyclic structure of a ring) with an amide acetal. The reaction of such amide acetals with primary amines is well known in the art, and those skilled in the art would be familiar with how to carry out the reaction with mitomycin C, mitomycin A or their N ^1a alkyl derivatives. Let's hang out. US Pat. No. 3,121,084 to HEWinberg (February 11, 1964) and R.
F. Abdulla et al., “The Chemistry of Formamide
Acetals”, Tetrehedron, Vol.35pp.1720−24
(1979) as a reference. Preferably, the reaction is carried out in a liquid, anhydrous reaction medium in which the diluent is a liquid compatible with the reaction conditions. Preferably the diluent is a lower halogenated aliphatic hydrocarbon or a lower alkanol or desirably a mixture of both. Chloroform and methanol and mixtures of both are highly suitable. The reaction is carried out at a temperature of 40° to 65°C for a sufficient period of time until the reaction is complete.
When a large excess (~60 times) of acetal is used, the major product formed is a bis-amidino product, a material of formula () where both A and B are amidino groups. N ^1a -formyl derivatives were sometimes formed as by-products. However, at moderate amounts (~10 times) of acetal, in addition to the bis-amidino product, a monoamidino product was also produced, a product of formula () where A is an amino group and B is an amidino group. From a mixture of these reaction products, individual products can be easily separated by chromatography, as described in the Examples below. [Table] Formula () in which R ² is cyano, di-lower alkylamino, lower alkoxy, or lower alkylthio
The material is prepared by replacing the amide acetal with the following orthocarbonate derivative in the method described above. These reagents are available according to the following literature: …Kantlehner, Liebigs Ann.Chem., 1981 ,
70−84. ,…H.Meerwein, Liebigs Ann.Chem.
641, 1 (1961). Amidino derivatives of formula () in which A is [Formula] or [Formula] are prepared from the anionic form of mitomycin C or its N ⁷ -substituted derivatives in the manner described above. Suitable halomethyleniminium salts for use in this process have been described in the literature; representative ones include:
“Advances in Organic Chemistry”, Vol.9,
Part 2, Wiley Interscience, 1979, pp.81and82.
Table 2 is quoted from the summary of the same document. [Table] [Table] When using halomethyleniminium salts such as those exemplified in Table 2, it is sometimes convenient to use the corresponding amide, ie the amide from which the iminium salt was prepared, as the solvent. If the corresponding amide is a solid, hexamethylphosphoramide or pyridine can be used. This was illustrated in Examples 17 and 19. Imidoyl chlorides derived from N-substituted formamides are also convenient reagents for this purpose. The manufacturing method is well established in the art and is illustrated in Table 3. Table 3 shows H. Ulrich “The
Chemistry of Imidoyl Halides”，Plenum
Press, New York, 1968, pp. 74-76. The reaction of imidoyl chloride with amines to form amidines is also well established, and SR
Sandler W. Karo “Organic Chemistry”, Vol.12
−, AT Blomquist and H. Wasserman, eds.
Academic Press, New York, 1972, p.227. [Table] [Table] Substances of formula () where A is a nitrogen-unsubstituted amidino group, [formula] or [formula] are amino-protected imino ethers and mitomycin C,
N ⁷ -substituted derivatives thereof, or any N ^1a -
It is produced by reaction with a lower alkyl derivative in anionic form as described above. Next, the protecting group is removed by a known method. Isopropylformimidate whose amino group is protected with a β-trimethylsilylethoxycarbonyl group is a suitable reagent (Example 13). (CH ₃ ) ₂ CHOCH=NH・HClClCO ₂ CH ₂ CH ₂ Si(CH ₃ ) ₃ ―――――――――――――――― (CH ₃ ) ₂ CHOCH=NCO ₂ CH ₂ CH ₂ When Si(CH ₃ ) ₃ A is (1-lower alkyl-2(1H)-pyridinylidene)amino or a group of the formula [Formula] or [Formula], the anionic form of mitomycin C is represented by the formula [Formula]. A production method is used in which R ² and R ³ of the compound are reacted with a cyclic halomethyleniminium salt or imidoyl halide in which R 2 and R 3 are bonded to form a ring. A suitable reagent for reaction with mitomycin C anion is 2-chloro-
1-Methylpyridinium iodide (Example
16), 2-chloro-4,5-dihydro-1-methyl-1(3H)-pyrrolidinium chloride (Table 2), N,N'-dimethyl-N,N'-trimethylene-chloroformamidinium chloride (Example
28) and 2-azetedinone, 2-pyrrolidinone,
Other cyclic imide halides derived from 2-piperidinone and 2-azepinone. Further, when R ⁷ or R ⁹ of the final product is hydrogen, a protecting group as described above is used in the intermediate cyclic halomethyleniminium salt. A substance of the formula () in which A or both A and B are amidino groups of the formula R ⁵ NH
primary amine (where R ⁵ is C _1-18 alkyl other than tertiary alkyl, C _1-18 alkenyl, C _1-18 alkynyl, C _1-18 haloalkyl, C _1-18 halooxyalkyl, C _{4 -8} cycloalkyl, or aryl or lower aralkyl each having up to 12 carbon atoms, or among heteroalicyclic or heteroaromatic groups having 3 to 8 membered rings, at least two of which are carbon atoms. reacts with The only limitations on the selection of the primary amine, other than the absence of functional groups that would conflict with the reaction conditions, are that the amino nitrogen atom must contain at least one hydrogen atom and that two or more aryl groups must be present. This means that it is bonded to a carbon atom that does not have a carbon atom. Anhydrous liquid organic compounds are used as the reaction medium; any material can be used as long as it is reasonable for the reaction conditions and does not participate in the reaction in a detrimental way. An excess of primary amine reactant on a molecular basis is generally used. Reaction temperatures of about -15°C to +50°C are preferred. The product of this reaction is 7-substituted amino-9α-
Methoxymitozane, i.e. on the 7-amino group
It is a mitomycin C derivative with a substituent as defined above for R ⁵ . Such compounds are known from the prior art to have a substantial degree of antitumor activity in experimental animals. It has been found that primary amines of the formula R ⁶ NH ₂ cannot be substituted with a 7-amidino group by the method described in the above section. In this R ⁶ , the carbon atom having an amino group is a tertiary carbon atom or 2
an alkyl, cycloalkyl, cycloalkyl, alkylaralkyl, or heteroalicyclic group having 4 to 18 carbon atoms which are secondary carbon atoms having 5 allyl groups. Weakly basic aliphatic amines such as trifluoroethylamine also cannot exchange the 7-amidino group. These amines are useful for converting compounds of formula (2), in which both A and B are said amidino groups, to compounds of formula (2), in which only A is said amidino group. Although these amines do not have the ability to replace the 7-amidino group, they can cleave the amidino group, denoted B, to NH ₂ to provide the carbamide functionality of unsubstituted mitozane. The amine itself can also act as a reaction medium or solvent system as defined in the previous section. This process is carried out at a reaction temperature range of 20°C to 60°C. The measurement method in the following examples is summarized. Melting points were determined on a Thomas-Hoover capillary melting point apparatus (no corrections were made). Temperatures are shown in degrees Celsius. Proton nuclear magnetic resonance spectra (hereinafter abbreviated as "NMR") were performed using a Varian XL100 spectrometer, using pyridine- _d5 unless otherwise noted.
(hereinafter abbreviated as "pyridine- _d5 ").
Infrared spectra (hereinafter abbreviated as "IR") were measured on samples compressed into KBr tablets using a Beckman 4240 spectrophotometer.
The IR numbers are ν _nax in cm ^-1 . Ultraviolet (hereinafter abbreviated as "UV") and visible spectra were measured with a Varian-Cary spectrophotometer. Using ultraviolet (UV) light as a measurement method,
Thin layer chromatography (hereinafter abbreviated as "TLC") was performed on a 0.25 mm precoated silica gel plate. Silica Woelm (32−
Flash chromatography was performed using 63 μm)]. The solvent was evaporated under reduced pressure below 50°C. Example 1 Compound 7 [(dimethylamino)methylene]amino-N ¹⁰ -(dimethylamino)-methylene-9a-
Methoxymitozane compound 7-[(dimethylamino)methylene]amino-N ¹⁰ -(dimethylamino)-methylene-
N ^1a -formyl-9a-methoxymitozane A total of 9.6 ml (0,
At 18, 21, and 23 hours, 2.4 ml of N,N-dimethylformamide dimethyl acetal was added and the suspension was stirred at about 50° C. for 41 hours. Evaporation of the solvent and excess reagent under reduced pressure gave a dark greenish-green residue. tel (methylene chloride/methanol 20:1)
The absence of mitomycin C and two types of green new components were confirmed (Rf=0.16 and 0.22). The main component (Rf=0.16) was isolated as a green solid (340 mg, 51.5%) by flash chromatography using methylene chloride/methyl 20:1 as eluent. This was dissolved in dimethyl ether and hexane was added to give the compound a dark green amorphous powder. NMR (pyridine d _-5 , δ); 2.18 (s, 3H), 2.70
(bs, 1H), 2.76 (s, 3H), 2.82 (s, 3H),
2.86 (s, 6H), 3.22 (s, 3H), 3.30 (bs, 1H),
3.60 (d, J=12Hz), 4.12 (dd, 1H, J=10,
4Hz), 4.43 (d, 1H, J=12Hz), 4.90 (bs,
1H), 5.10 (t, 1H, J=10Hz), 5.52 (dd,
1H, J=10, 4Hz), 7.85 (s, 1H), 8.64 (s,
1H). IR (KBr) ν _nax ′cm ^-1 : 3300, 2930, 1675, 1620,
1545, 1230, 1060. UV( _H2O )λ _nax′nm : 390, 244. Elemental analysis: ( _C21H28N6O5 ): Calculated: C, 56.71; H, 6.08 _: N, 18.90 Measured _{: C} , 56.20; H, 6.28; N, 17.88. A minor component (Rf = 0.22) of (180 mg, 25.35%) isolated as a non-crystalline solid precipitated from dimethyl ether and hexane was identified as the compound. NMR (pyridine d ₅ , δ): 2.20 (s, 3H), 2.60
-3.00 (3 singlets, 12H), 3.2 (s, 3H),
3.65 (m.2H), 4.04 (d, 1H, J=4Hz), 4.16
(dd, 1H, J = 12, 4Hz), 4.60 (d, 1H, J
= 13Hz), 4.86 (t, 1H, J = 12Hz), 4.90 (s,
1H), 5.48 (dd, 1H, J=12, 4Hz), 7.90 (s,
1H), 8.64 (s, 1H), 9.06 (s, 1H). IR (KBr) ν _nax ′cm ^-1 : 2490, 2860, 1698, 1630,
1600, 1540, 1250, 1060. UV ( _H2O ), λ _nax′nm : 390, 244. Elemental analysis: ( _C22H25N6O6 ): Calculated: C, 55.89; H, 5.93; _N , 17.78 Observed _: C _, 55.41; H, 5.96; N, 16.99. When a solution of the compound in ethyl acetate or N,N-dimethylformamide dimethyl acetal is allowed to stand at room temperature for more than 10 hours, the compound (Rf=
TLC revealed that 0.22) had been converted into a compound (Rf=0.16), resulting in a solution extremely rich in the latter. Examples 2-7 are Example 1 with the following modifications to synthesize various other compounds of the present invention.
This method was implemented. Example 2 Compound 7 - [(diisopropylamino)methylene]amino- ^N10- (diisopropylamino)-methylene-9a-methoxymitozane N,N-diisopropylformamide Mitomycin C in diethyl acetal (3 ml)
(200mg, 0.6mM) suspension for 15 hours with stirring.
Heated to 53°C. Pour the reaction mixture into 50ml of water,
Extracted with ethyl acetate (3x30ml). The combined organic extracts were dried (Na ₂ SO ₄ ) and evaporated to a dark green syrup. Rapidly moving impurities (Rf = 0.45 ~
It became clear that there was a green principal component (Rf = 0.43) with an Rf of 0.50). The main component was a dark green solid (156 mg, 46.8%) in methylene chloride/methanol by flash chromatography twice.
20:1 was used as the eluent. NMR ( _CDCl3 , δ): 1.10-1.50 (5 singlets, 24H), 1.94 (s, 3H), 2.78 (dd, 1H, J
= 4, 2Hz), 3.05 (d, 1H, J = 4Hz), 3.22
(s, 3H), 3.60 (m, 5H), 3.75 (dd, 1H, J
= 10, 4Hz), 4.24 (d, 1H, J = 12Hz), 4.56
(t, 1H, J=10Hz), 4.88 (dd, 1H, J=10,
4Hz), 7.83 (s, 1H), 8.67 (s, 1H). IR (KBr), ν _nax ′cm ^-1 : 3320, 2990, 2940,
1680, 1630, 1600, 1550, 1235, 1060. UV(MeOH)λ _nax′nm : 246, 393. Elemental analysis: ( _C29H44N6O5 ): Calculated: C, 62.55; H, 7.91; _N , 15.10 Measured _{: C} , 62.03; H, 7.80; N, 14.60. Example 3 Compound 7-[(dimethylamino)methylene]amino-
N ¹⁰ -(dimethylamino)-methylene-9a-methoxy-N ^1a -methylmitozane In this example, 10 ml of chloroform and 2 ml
methanol was used as the reaction solvent, 0.8ml
The starting material reacting with N,N-dimethylformamide dimethyl acetal (1.5 mM) was replaced with 130 mg (0.37 mM) of porfiromycin (N ^1a -methyl mitomycin C).
The reaction time was 50 minutes at 50°C. The compound was obtained as a syrup on evaporation of the reaction solvent. 20g silica gel and methylene chloride/methanol (20:
This was purified by flash chromatography using 1) as the eluent. NMR (pyridine d ₅ , δ): 2.22 (bs, 4H),
2.28 (s, 3H), 2.70 (d, 1H, J=4Hz),
2.80 (s, 3H), 2.84 (s, 3H), 2.90 (s, 6H),
3.20 (s, 3H), 3.52 (dd, 1H, J=2, 12Hz),
4.10 (dd, 1H, J=4, 11Hz), 4.38 (d, 1H,
J = = 12Hz), 4.92 (t, 1H, J = 11Hz), 4.96
(bs, 1H), 5.46 (dd, 1H, J=4, 11Hz),
7.86 (s, 1H), 8.70 (s, 1H). Rf=0.53 (methylene chloride/methanol 9:1
tlc). IR (KBr) ν _nax ′cm ^-1 : 2930, 1680, 1620, 1545,
1230, 1115. UV(MeOH)λ _nax′nm : 386, 243. Elemental analysis: ( _C22H30N6O5 ): Calculated: C, 57.60; _H , 6.55; _N , _18.33 Measured: C, 57.11; H, 6.11; N, 17.90. In this method, the compound 7-amino- ^N10 -dimethylamino-methylene-9a-methoxy- ^N1a-
Methylmitozane was produced as a by-product in 30% yield. tlc Rf = 0.40 (methylene chloride/methanol 9:1). NMR (pyridine d ₅ , δ): 2.02 (s, 3H),
2.16 (dd, 1H, J=2.5Hz), 2.25 (s, 3H),
2.66 (d, 1H, J=5Hz), 2.76 (s, 3H),
2.86 (s, 3H), 3.18 (s, 3H), 3.51 (dd, 1H,
J=2, 12Hz), 4.08(dd, 1H, J=4, 10
Hz), 4.50 (d, 1H, J=10Hz), 4.90 (t, 1H,
J=10Hz), 5.05(bs), 5.43(dd, 1H, J=4,
10Hz), 8.70 (s, 1H). IR (KBr) ν _nax ′cm ^-1 : 3430, 3330, 3270, 2940,
2960, 1690, 1625, 1553, 1230, 1125. UV(MeOH)λ _nax′nm : 358, 244, 216. Elemental analysis: ( _C19H25N5O5 ): Calculated: C, 56.53; H, 6.20; _N , 17.38 Measured: C _, 54.68; _H , 6.13; N, 16.59. Example 4 Compound 9a-Methoxy-7-[(1-piperidinylamino)methylene]amino- ^N10- (1-piperidinylmethylene)mitozane N-(diethoxymethyl)piperidine in chloroform (3 ml) solution 3ml and mitomycin
200 mg of C was reacted at 60°C for 2 hours. The product is 27.6
% yield. tlc Rf = 0.20 (methylene chloride/methanol 20:
1) NMR (pyridine d ₅ , δ): 1.38 (bs, 12H),
2.20 (s, 3H), 2.80 (bs, 1H), 3.24 (s, 3H),
3.00-3.40 (m, 5H), 3.40-3.80 (m, 5H),
4.13 (dd, 1H, J=10, 4Hz), 4.45 (d, 1H,
J = 12Hz), 4.90 (bs, 2H), 5.12 (t, 1H, J
= 10Hz), 5.56 (dd, 1H, J = 10, 4Hz), 7.87
(s, 1H), 8.70 (s, 1H). IR (KBr) ν _nax ′cm ^-1 : 3300, 2950, 2870, 1680,
1630, 110, 1550, 1200, 1070. UV( _H2O )λ _nax′nm : 394, 246. Elemental analysis: ( _C27H36N6O5 ): Calculated: C, 61.79; H, 6.87; _N , 16.02 Measured _{: C} , 61.01; H, 6.85; N, 15.34. N ^1a -formyl derivatives of the aforementioned substances, compounds
N ^1a -Formyl-9a-methoxy-7-[(1-piperidinylamino)methylene]amino-N ¹⁰ -
(1-piperidienylmethylene)mitozane was obtained as the main component in a yield of 43%. tle Rf=0.25 (methylene chloride/methanol 20:1) NMR (pyridine d ₅ , δ): 1.38 (bs, 12H),
2.23 (s, 3H), 3.00-3.40 (m, 4H), 3.23 (s,
3H), 3.40-3.90 (m, 6H), 4.07 (d, 1H, J
= 4Hz), 4.18 (dd, 1H, J = 11, 4Hz), 4.63
(d, 1H), 4.0 (t, 1H, J=11Hz), 4.94 (bs,
1H), 5.54 (dd, 1H, J=11, 4Hz), 7.94 (s,
1H), 8.71 (s, 1H), 9.08 (s, 1H). IR (KBr) ν _nax ′cm ^-1 : 2490, 2860, 1698, 1630,
1600, 1540, 1250, 1060. UV( _H2O )λ _nax′nm : 394, 247. Elemental analysis: ( _C28H36N6O6 ): Calculated: C, _60.08 ; H, 6.52; N, 15.21 Measured _: C _, 59.99; H, 6.17; N, 15.07. Example 5 Compound 9a-Methoxy-7-[(1-morpholino)methylene]amino- ^N10- (1-morpholino)-methylenemitozane Mitomycin C in chloroform (10 ml) and N-dimethoxymethyl morpholine (4 ml)
(200mg, 0.6mM) was heated at approximately 53°C for 42 hours. The reaction mixture was concentrated to a syrup under high vacuum. The greenish component was separated from the excess reagent by crude flash chromatography in (methylene chloride/methanol 25:1). The combined green components were dissolved in 20 ml of ethyl acetate and washed with water (3
The washings were combined and re-extracted with ethyl acetate (3 x 15 ml). Combine all ethyl acetate moieties,
It was dried (Na ₂ SO ₄ ) and evaporated to a dark green syrup. This tlc (methylene chloride/methanol 10:
In 1), a green component of Rf=0.33 with several green impurities (Rf=0.35-0.40) was clearly confirmed. The compound was identified as a dark green amorphous solid (130
mg, 56.8%). NMR (CDCl ₃ , δ): 1.91 (s, 3H), 2.80 (bs,
1H), 3.13 (d, 1H, J=2Hz), 3.22 (s,
3H), 3.30−3.94 (m, 18H), 4.20 (d, 1H, J
= 12Hz), 4.40 (bs, 1H), 4.54 (t, 1H, J =
10Hz), 4.88 (dd, 1H, J=10Hz, 4Hz), 7.74
(s, 1H), 8.51 (s, 1H). IR (KBr) ν _nax ′cm ^-1 : 3300, 2970, 2920, 1680,
1625, 1550, 1235, 1070. UV(MeOH)λ _nax′nm : 386, 244. Elemental analysis: ( _C22H32N6O7 ): Calculated: C, 56.78; _H , 6.06; _N , 15.90 Measured _: C, 53.07; H, 6.03; N, 15.37. Example 6 Compound 7-amino- ^N10 -dimethylaminomethylene-
9a-Methoxymitozane Dissolve mitomycin C (200 mg, 0.6 mM) in 10 ml chloroform and 2 ml methanol,
Add N,N-dimethylformamide dimethyl acetal (0..64ml, 4.8mM) and incubate at approximately 50°C.
Stir the solution for 50 minutes. Thin layer chromatography (methylene chloride/methanol 90:10) revealed traces of unreacted mitomycin C ((Rf = 0.22) and two new components (Rf = 042 and 0.33). The solution was syruped under reduced pressure. The syrup was separated by flash chromatography on (25 g silica gel) using methylene chloride/methanol (20:1) as the eluent. The rapid component (Rf = 0.42) was separated as a green amorphous solid ( 60
mg, 22.5%) and identified the compound by NMR spectrum (pyridine d- ₅ ).
The main component of blue color (Rf = 0.33) was converted into an amorphous solid (148mg,
63.3%) and confirmed that it was a compound. The analytical sample was n from a methylene chloride solution.
- Obtained by precipitation with pentane. NMR (pyridine d ₅ , δ): 2.02 (s, 3H),
2.76 (b, 4H), 2.86 (s, 3H), 3.21 (s, 3H),
328 (d, 1H, J = 4Hz), 3.62 (dd, 1H, J =
2, 13Hz), 3.94 (bs), 4.14 (dd, 1H, J=4,
12Hz), 4.56 (d, 1H, J=13Hz), 5.12 (t,
1H, J=10Hz), 5.52 (dd, 1H, J=4, 10
Hz). IR (KBr) ν _nax ′cm ^-1 : 3430, 3320, 3280, 2930,
1675, 1615, 1650, 120, 1115. UV( _H2O )λ _nax′nm : 364, 244, 219. Elemental analysis: (C ₁₈ H ₂₃ N ₅ O ₅ ): Calculated value: C, 55.48; H, 5.91; N, 17.98 Measured value: C, 54.70; H, 6.14; N, 17.95 Example 7 Compound 7,9a- Dimethoxy- ^N10 -dimethylaminomethylenemitozane N,N-dimethylformamide dimethyl in a chloroform/methanol (10:1) solution for 1 hour at 50°C using mitomycin A (170 mg) in place of mitomycin C in Example 1. Acetal (0.6ml)
I reacted. The desired product was obtained in 48%. tlc Rf
= 0.50 (methylene chloride/methanol 9:1). NMR (pyridine d ₅ , δ): 1.83 (s, 3H),
2.76 (bs, 4H), 2.86 (s, 3H), 3.22 (s, 3H),
3.28 (d, 1H), 3.56 (dd, 1H, J=2, 13Hz),
4.02 (s, 3H), 4.10 (dd, 1H, J=4, 10Hz),
4.24 (d, 1H, J = 13Hz), 5.10 (t, 1H, J =
10Hz), 5.50 (dd, 1H, J=4, 10Hz), 8.67
(s, 1H). IR (KBr) ν _nax ′cm ^-1 : 3300, 2930, 1675, 1655,
1625, 1500, 1235, 1120. UV( _H2O )λ _nax′nm : 530, 316, 244. Elemental analysis: ( _C19H24N4O6 ): Calculated: C, 56.39; H, 5.94; _N , 13.85 Measured _: C, 56.51; _H , 5.92; N, 13.71. The N ^1a -formyl derivative of the compound is 16.5% as the compound, 7,9a-dimethoxy-N ¹⁰ dimethylaminomethylene-N ^1a -formyl mitozane.
Obtained in yield. tlc Rf = 0.61 (methylene chloride/methanol 9:1) NMR (pyridine d ₅ , δ): 1.88 (s, 3H),
2.76 (s, 3H), 2.85 (s, 3H), 3.54 (d, 1H),
3.62 (bs, 1H), 4.05 (s, 3H), 4.05 (bs, 1H),
4.14 (dd, 1H, J=4, 12Hz), 4.40 (d, 1H,
J=13Hz), 4.86 (t, 1H, J=12Hz), 5.42
(dd, 1H, J=4, 12Hz), 8.66 (s, 1H),
9.08 (s, 1H). Example 8 Compound 7-(dimethylaminomethylene)amino-9a-
Amidinophenylmethane (2.2ml, 10.8mM) was added to the compound (600mg, 1.35mM) dissolved in methoxymitozane methanol (10ml) and the resulting solution was
Stirred at ℃ for 4 hours. The progress of the reaction was monitored by tic (methylene chloride/methanol 90:10). After 4 hours, the starting material (Rf = 0.35) had disappeared and in its place a main green band (Rf = 0.29) had formed. The solution was concentrated under reduced pressure and the resulting syrup was subjected to flash chromatographic analysis using methylene chloride/methanol 20:1 as eluent (25 g silica gel). Green component (Rf=
0.29) are collected and dried (Na ₂ SO ₄ ),
Concentrated. The compound is an obligatory crystalline solid (215 mg, 41
%). NMR (pyridine d ₅ , δ): 2.18 (s, 3H),
2.70 (bs, 1H), 2.80 (s, 3H), 2.88 (s, 3H),
3.08 (bs, 1H), 3.24 (s, 3H), 3.56 (bd, 1H,
J = 12Hz), 4.00 (dd, 1H), 4.44 (d, 1H, J
= 12Hz), 5.06 (t, 1H, J = 10Hz), 5.56 (dd,
1H, J=10, 4Hz), 7.58 (bs, 2H), 7.88 (s,
1H). IR (KBr) ν _nax ′cm ^-1 : 3300−3450, 2960−2910,
1715, 1620, 1535, 1050. UV( _H2O )λ _nax′nm : 390, 226. Elemental analysis: ( _C18H23N5O5 ): Calculated: C, 55.48; H, 5.91; _N , 17.98 Measured _: C _, 54.83; H, 5.67; N, 16.90. N ^1a in place of the starting material for the compound in Example 8
- formyl derivatives, using compounds at room temperature for 20
When reacted for hours, the compounds were obtained in virtually the same manner and yield. Example 9 Compound 7-(dimethylaminomethylene)amino-9a-
Methoxy- ^N1a -methylmitozane compound 1g (2.18mM) was dissolved in methanol (20ml), aminodiphenylmethane (3.5ml, 17.18mM) was added and the resulting solution was heated at room temperature for 5 hours and at 40°C. The mixture was stirred for 5 hours. Thin layer chromatography of the reaction mixture (CH ₂ Cl ₂ /
MeOH (90:10), most of the starting material (Rf = 0.55) had disappeared, and a new green main component (Rf = 0.48) was observed to have been produced. The compound was prepared as an amorphous solid (350 mg) using a similar procedure as in Example 8.
obtained as. Purification was performed using CH ₂ Cl ₂ /MeOH (250 ml,
Analytically pure samples were obtained by flash chromatography (7 g silica gel) using 96/4 v/v) and precipitation of the resulting solid (Rf = 0.48) from methylene chloride (5 ml) and hexane (50 ml). (314 mg, 35.7%) as a solid. NMR (CDCl ₃ , δ): 1.93 (s, 3H), 2.26 (bs,
1H), 2.26 (s, 3H), 3.06 (s, 3H), 3.08 (bs,
1H), 3.10 (s, 3H), 3.20 (s, 3H), 3.46
(bd, 1H, J = 12, 1Hz), 3.58 (dd, 1H, J
= 4, 10Hz), 4.17 (d, 1H, J = 12Hz), 4.38
(t, 1H, J=10Hz), 4.68 (m, 2H), 4.76
(dd, 1H, J=4.10Hz), 772 (s, 1H). IR (KBr) ν _nax ′cm ^-1 : 3440, 3350, 3190, 3020,
2940, 210, 1725, 1630, 1550, 1055. UV(MeOH)λ _nax′nm : 386, 231. Elemental analysis: ( _C19H25N5O5 ): Calculated: C, 56.53; H, 6.20; _N , 17.36 Measured _: C _, 53.90; H, 5.13; N, 15.81. Example 10 Compound XI 7-(n-propyl)amino-9a-methoxymitozane Compound (330 mg,
Dissolve n-propylamine (1.0
ml) was added. The reaction mixture was stirred at room temperature for 6 hours and at about 0°-4° for 16 hours. The solvent and excess reagents were evaporated under reduced pressure, and the residue was subjected to flash chromatographic analysis using silica gel as an adsorbent. The blue component (Rf = 0.40) obtained in a solution of methylene chloride/methanol 30:1 was reprecipitated from methylene chloride with hexane to give compound XI as an amorphous gray powder (125 mg, 44.5%). NMR (pyridine d ₅ , δ): 0.80 (t, 3H),
1.42 (m, 2H), 2.11 (s, 3H), 2.74 (bs, 1H),
3.12 (bs, 1H), 3.22 (s, 3H), 3.36 (q, 2H),
3.60 (d, 1H, J = 12Hz), 3.96 (dd, 1H, J =
11Hz, 4Hz), 4.54 (d, 1H, J=12Hz), 5.00
(m, 3H), 5.36 (dd, 1H, J=11, 4Hz),
6.90 (t, 1H). IR (KBr) ν _nax ′cm ^-1 : 3400, 3300, 2960, 2940,
1715, 1630, 1600, 1550, 1510, 1220, 1060. UV( _H2O )λ _nax′nm : 372, 222. Elemental analysis: ( _C18H24N4O5 ): Calculated: C, 57.40; H, 6.38; _N , _14.88 Measured _: C, 57.28; H, 6.41; N, 14.08. Example 11 Compound XII 7-(2-hydroxyethyl)amino-9a-methoxymitozane Compound (330 mg,
0.74mM) and ethanolamine (2ml).
was added. The reaction mixture was stirred at room temperature for 2 hours,
Diluted with water (50ml) and extracted with ethyl acetate (5x60
ml). The combined ethyl acetate extracts were dried (Na ₂ SO ₄ ) and concentrated to a blue-purple residue, which was concentrated by column chromatography using 10% methanol in methylene chloride, containing the blue component. 105 mg of compound XII as an amorphous solid from the fraction
(37%). NMR (pyridine d ₅ , δ): 2.14 (s, 3H),
2.81 (bs, 1H), 3.18 (d, 1H, J=4Hz),
3.24 (s, 3H), 3.65 (dd, 1H, J=2, 12Hz),
3.70−4.20 (m, 5H), 4.52 (d, 1H, J=13
Hz), 4.96 (t, 1H, J=12Hz), 7.38 (t,
1H), 7.58 (bs). IR (KBr) ν _nax ′cm ^-1 : 3300−3500, 2930, 1710,
1630, 1600, 1540, 1510, 1200, 1055. UV( _H2O )λ _nax′nm : 371, 221. Elemental analysis: ( _C72H22N4O6 ): Calculated: C, 53.92; H, 5.82; _N , 14.80 Measured _: C, 51.30; _H , 5.88; N, 14.80. Example 12 Compound 7-[2-benzylthioethyl]amino-9a-
Methoxymitozane Compound (200 mg,
S-benzyl 2-aminoethanethiol (0.5ml) was added and the solution was stirred at room temperature for 16 hours. The residue obtained by evaporation of the solvent under reduced pressure was dissolved in 6% methanol/methylene chloride (400 ml).
It was analyzed by flash chromatography (40 g, silica gel) using as eluent. The blue component (Rf ~0.5, 10% MeOH/CH ₂ Cl ₂ ) was liberated as an amorphous solid (65 mg, 29.8%). Its spectral data (NMR, IR, UV and mass spectrometry)
matched the planned structure. Elemental analysis: ( _C24H26N4O5S ) _Calculated : C, 59.49; H, 5.82; _N , 11.56 Measured _: C, 59.72; H, 5.94; N, 11.08. Example 13 Preparation of Formula (A) Diisopropylethylamine (2.1 mM) was slowly added to a solution of isopropylformimidate hydrochloride (1 mM) in 2 ml of dimethylformamide (DMF) at 0° C. under nitrogen atmosphere. β-Trimethylsilylethyl chloroformate was added dropwise to the resulting solution at 0°C. The resulting clear solution is referred to as solution A. (B) Mitomycin C (1mM) in 5ml DMF
To the solution was added a suspension of sodium hydride (1.5mM) in 2ml of DMF. Solution at room temperature 20
Stir for -40° to -50°C before adding solution A.
It was cooled to The solution was kept at -40°C for 1 hour and then allowed to warm up to room temperature. After standing at room temperature for about 6-18 hours, the reaction mixture was diluted _{with CH2Cl2} and filtered. The solid residue obtained on evaporation of the liquid was chromatographed on silica gel to separate the amidino-protected title compound. (C) The amidino-protecting group of the above-mentioned intermediate is the same as that of Carpino and Tsao (J.Chem.Soc.Chem.Comm.358 (1978)).
to give the unsubstituted amidino title compound. Example 14 Preparation of Formula (A) Diisopropylethylamine (21 mM) was slowly added to a solution of isopropylformimidate hydrochloride (1 mM) in DMF (2 ml) under a nitrogen atmosphere at 0°C. Methyl iodide was added to the resulting solution at 0°C. This is called solution B. (B) The method described in Example 13(B) was carried out using Solution B in place of Solution A to obtain the title compound. Example 15 A 0.5M N,N-dimethylchloromethylreniminium chloride solution was added to 25 ml of CHCl ₃ at 0°C.
Oxalyl chloride (1.57g, 12.5mM) was added dropwise to a DMF (915mg, 12.5mM) solution in the solution and then stirred for 30 minutes at room temperature. separately
Mitomycin C (334mg, 1mM) in 5ml DMF
To the solution was added 3 ml of a suspension of NaH (36 mg, 1.5 mM) in DMF. The solution was stirred at room temperature for 20 minutes, then cooled to -40° to -50°C and treated with N,N-dimethylchloromethyleniminium chloride (3
ml, 1.5mM) of the above solution was added. 10 at −40℃
After stirring for a minute, additional NaH (18 mg, 0.75 mM) was added. The solution was kept at −40°C for 1 hour, then
Diluted with CH ₂ Cl ₂ and filtered. The residue obtained by evaporation of the liquid was dissolved in silica gel (10% CH ₃ OH−
Chromatographic analysis was performed by thin layer chromatography using CH ₂ Cl ₂ as eluent. The extract of the main green zone is 78 mg (43 mg based on recovered mitomycin C).
%) of an amorphous solid whose NMR spectrum and TLC behavior were synthesized in Example 8. It was the same as the compound. The purple band extract was 150 mg mitomycin C. Example 16 7-(1-methyl-2-(H)-pyridinylidene)
Amino-9a-methoxymitozane Mitomycin C (242mg, 0.725mM) and
4ml DMF in a mixture of NaH (43.5mg, 1.81mM)
added. After stirring for 15 minutes, 2-chloro-1-
Methylpyridinium iodide (370mg,
1.45mM) was added at room temperature. The solution was stirred for 1.5 hours, diluted with ethyl acetate (EtOAc) and filtered. The residue obtained by evaporation of the liquid was purified on silica gel (5
% _{CH3OH -- CH2Cl2} as eluent) (tlc) chromatographic fractionation. A minor component (12 mg) was the compound (Example 8). The main product (75 mg) was further purified by silica gel TLC (10% CH ₃ OH-CH ₂ Cl ₂ ) to give 6 mg (2%) of the title compound. NMR (pyridine d ₅ , δ): 2.11 (s, 3H),
2.76 (bs, 1H), 3.20 (m, 1H), 3.26 (s, 3H),
3.49 (s, 3H), 3.63 (dd, 1H, J=13, 1Hz),
4.01 (dd, 1H, J=11, 4Hz), 4.51 (d, 1H,
J = 13Hz), 5.10 (t, 1H, J = 10Hz), 5.43
(dd, 1H, J = 10, 4Hz), 5.99 (dt, 1H, J
= 9, 2Hz), 6.09 (dd, 1H, J = 9, 1Hz),
6.95 (dd, 1H, J=9, 7, 2Hz), 7.32 (dd,
1H, J=7, 1Hz). Example 17 7-[(methylaminomethylene)amino]-9a-
Methoxymitozane To a solution of mitomycin C (167 mg, 0.5 mM) in 2 ml of hexamethylphosphoramide was added sodium hydride (12 mg, 0.5 mM) under a nitrogen atmosphere. Add N-methylformimidoyl chloride (19mg, 0.25mM NHBosshard) to this solution.
and H. Zollinger, Helv. Chim. Acta, 42 , 1659.
(1959)) was added. The solution was stirred at room temperature for 10 minutes, then added with NaH (6 mg, 0.25 mM) and N-methylformimidoyl chloride (9.5 mg,
0.13mM) was added. After stirring for 6-12 hours, the solution was diluted with ethyl acetate and filtered. Evaporation of the solvent and chromatographic purification of the residue gave the title compound. Example 18 Compound XI 9a-methoxy-7-(1-morpholinomethylene)aminomitozane 4-Diethoxymethyl morpholine (12.5 ml) was added to mitomycin C (600 mg, 1.8 mM) suspended in chloroform (30 ml), and the resulting suspension was diluted to 58
Heated at ℃ for 48 hours. TLC (20% MeOH in CH ₂ Cl ₂ ) reaction after 48 hours was found to be insufficient. The solution was concentrated under reduced pressure and the resulting syrup was added to water (100ml). After stirring for 20 minutes, the dark green solution was extracted with methylene chloride (5 x 50ml).
The combined extracts were dried and concentrated to syrup. To this syrup was added aminodiphenylmethane (6.5ml) in methanol (20ml) and the resulting solution was stirred at 30-35°C for 18 hours. Thin layer chromatography (20% MeOH in CH ₂ Cl ₂ ) revealed a green main component band with a slight purple band. The solution was concentrated under reduced pressure, and the resulting syrup was purified by conventional flash chromatography to obtain the title compound as a dark green amorphous solid (75
mg, 10%). Analytical samples were obtained by precipitation with n-hexane from a methylene chloride solution. NMR (pyridine d ₅ , δ): 2.16 (s, 3H),
2.76 (dd, 1H, J=5, 1Hz), 3.16 (d, 1H,
J = 5Hz), 3.24 (s, 3H), 3.28-3.80 (m,
10H), 4.02 (dd, 1H, J=10, 4Hz), 4.04
(d, 1H, J=12Hz), 5.06 (t, 1H, J=10
Hz), 5.46 (dd, 1H, J=10, 4Hz), 7.90 (s,
1H). IR (KBr) ν _nax ′cm ^-1 : 3360, 3280, 2960, 2920,
1720, 1600, 1520, 1230, 1050. UV(MeOH)λ _nax : 384, 234. Elemental analysis: ( _C20H25N5O6 ) _Calculated : C, 55.64; _H , 5.80; N, 16.23 Measured _: C, 55.07; H, 5.55; N, 15.88. Example 19 Compound XII 7-(1-pyrrolidinylmethylene)amino-
9a-Methoxymitozane Oxalyl in a solution of 1-formylpyrrolidine (2.48 g, 25 mM) in 50 ml of CHCl ₃ at 0 °C.
Chloride (3.17 g, 25 mM) was added dropwise and stirred at room temperature for 30 minutes to prepare a 0.5 molar solution of pyryridinylchloromethyleneium chloride.
Separately, sodium hydride (24 mg, 1 mM) was added to a solution of mitomycin C (334 mg, 1 mM) in 3 ml of 1-formylpyrrolidine under a nitrogen atmosphere.
After stirring the solution at room temperature for 20 minutes, it was cooled to -40° to -50°C and added with the iminiim salt solution prepared above (1 ml,
0.5mM) was added. Add 12mg (0.5mM) NaH, 0.5ml (0.25mM) to this mixture at 10 minute intervals.
iminium salt solution, 6 mg (0.25 mM) NaH,
0.25ml (0.125mM) iminium salt solution and finally 3mg (0.125mM) NaH and 0.125ml
(0.063mM) of iminium salt solution was added. After stirring at −30° C. for 30 minutes, the mixture was warmed to room temperature. It was diluted with ethyl acetate and the inorganic salts were separated.
The residue obtained after evaporation of the solvent was dissolved in silica gel (10%
Chromatographic fractionation was performed by thin layer _{chromatography} on _CH3OH -- _CH2Cl2 . The green band extract gave 120 mg (15% yield) of the title compound. NMR (pyridinc d ₅ , δ): 1.58 (m, 4H),
2.29 (s, 3H), 2.73 (m, 1H), 3.06−3.50 (m,
8H), 3.59 (dd, 1H, J=13, 1Hz), 4.03
(dd, 1H, J = 10, 4Hz), 4.44 (d, 1H, J
= 12Hz), 5.05 (t, 1H, J = 10Hz), 5.45 (dd,
1H, J=10, 4Hz), 8.04 (s, 1H). IR (KBr) ν _nax ′cm ^-1 : 3420, 3280, 2960−2870,
1715, 1625, 1560, 1300, 1055. Example 20 7-[N-methyl-N-(methylamino)methyl]amino-9a-methoxymitozane The method of Example 17 was repeated using a similar molecular weight of 9a-methoxy-7-(N-methylamino)mitozane (Matsui et al., The
Journal of Antibiotics, XI, 189-198
(1968)). Example 21 Compound 7-[1-(dimethylamino)ethylidene]amino- ^N10- [1-(dimethylamino)ethylidene]-9-methoxymitozane 600 mg mitomycin in 2 ml methanol
(1.79mM) was prepared and treated with 3ml of N,N-dimethylacetamide dimethylacetal. The suspension was heated to 75-80°C for 2 hours with stirring. Almost all mitomycin C was removed by tlc (CH ₂ Cl ₂ /methanol 10:1) at this step.
It showed that the amount was consumed by the reaction. The product appeared as a green band. The reaction mixture was concentrated to dryness under reduced pressure to obtain a syrup free of solvent and volatiles. This was dissolved in methylene chloride and packed into a silica gel column (40g silica gel), and the column was filled with 1% methanol in methylene chloride (200ml), 2% methanol in methylene chloride (200ml) and 5% methanol in methylene chloride. (400 ml). The fraction containing the product green band was collected and concentrated to 110 mg (13
% yield) of an amorphous solid was obtained. This material was dissolved in 2 ml of acetetone and precipitated from solution by the addition of hexane. The product was collected by filtration. Elemental analysis: ( _C23H32N6O5 ): Calculated: C, 58.46; H, 6.83; _N , _17.79 Measured: _C , 58.89; H, 6.89; N, 17.64. UV (MeOH) λ _nax゜nm: 235, 364. IR (KBr) ν _nax ′cm ^-1 : 3440, 3295, 2925, 1770,
1660, 1620, 1580, 1550, 1300, 1055. ^{The 1} H NM (pyridine d ₅ ) spectrum was consistent with the structure of the title compound. Example 22 Compound 7-[1-(dimethylamino)ethylideneamino]-9a-methoxymitozane 100 mg of compound in 2 ml chloroform
(0.21mM) of aminodiphenylmethane 2
ml was added and the solution was heated to approximately 55-60°C for several hours. Traces of compound remained in the reaction mixture at this stage. However, the concentrated residue was chromatographed on neutral alumina using a gradient elution starting with methylene chloride and ending with methanol/methylene chloride 2.5:1. 25 mg (29.4% yield) of the main component green band
It was isolated as an amorphous green solid. This material was dissolved in acetone and purified by adding hexane to the acetone solution until a precipitate formed. The product was collected by filtration. Elemental analysis: ( _C25H32N6O5 ) Calculated: C, _56.58 ; H, 6.20; N, 17.37 Measured _: C, 55.71; _H , 6.34; N, 15.23. UV (H ₂ O) λ _nax゜nm: 374, 230 (Shoulder). IR (KBr) ν _nax ′cm ^-1 : 3420, 3350, 3280, 2920,
1710, 1610, 1540, 1300, 1050. ^{The 1} H NMR (pyridine d ₅ ) spectrum was consistent with this structure. Example 23 Compound 7-[(1-methyl-2-pyrrolidinylidene)amino] ^-N10 -[(1-methyl-2-pyrrolidinylidene)amino-9a-methoxymitozane 2,2-dimethoxy-1 in 20ml methanol
-Methylpyrrolidine (H. Eilingsfeld. Angew.
Chem., 72 , 836 (1960)) 1.5g (10.3mM) and mitomycin C 280mg (0.34mM) at 55°C for 5 hours.
heated with. The reaction mixture was checked by TLC on an alumina plate in methylene chloride/methanol 97:3. TLC showed a green main component point, which is the product, and a blue point, which is the starting material mitomycin C. The solvent was removed by vacuum distillation at 40°C and the residue was dissolved in methylene chloride to give a solution consisting of 150 g of alumina.
cm column. Eluted with 50 ml of methylene chloride and 600 ml of 1% methanol in methylene chloride. Although most of the impurities were removed, no pure fraction was liberated. Combined eluent at 20℃
It was distilled and concentrated to give an oily residue. This includes 2,
It was clear that 2-dimethoxy-1-methylpyrrolidine was contained. This material was transferred again to an alumina column (25 g of alumina) in 200 ml of methylene chloride and 100 ml of 1% methanol in methylene chloride.
Chromatographic fractionation was performed. As a result, 2,2-dimethoxy-1-methylpyrrolidine was removed, and a fraction containing a small amount of impurities and a pure fraction were obtained. The pure fraction was confirmed to be the desired product by TLC (1 green dot). Yield 33mg. Elemental analysis: ( _{C25H32N6O5.0.85H2O} ) Calculated _values : C, _58.66 ; _H , 6.64; _N , 16.42. Measured values: C, 58.63; H, 6.46; N, 16.50. UV (MeOH) λ _nax゜nm: 354, 239. IR (KBr) ν _nax ′cm ^-1 : 3300, 3220, 2940,
11660, 1620, 1550, 1290, 1055. ^{The 1} H NMR (pyridine d ₅ ) spectrum was consistent with the structure of the title compound. Example 24 7-[(1-methyl-2-pyrrolidinylidene)amino]-9a-methoxymitozane 80mg of compound in 15ml chloroform
(0.16mM) and n-butylamine 0.48ml solution.
Heated under reflux for 48 hours. tlc (on 2% alumina,
Methanol/methylene chloride) appeared as the main component green dot and a small leading blue dot and a small trailing red dot that was part of the starting material. reaction solution
It was placed on a column containing 50 g of alumina and eluted with 200 ml of 1% methanol in methylene chloride and 400 ml of 2% methanol in methylene chloride. Fractions containing a single major green component that appeared on tlc were collected and concentrated to a residue of 24 mg of the desired product. NMR (pyridine d ₅ , δ): 1.72 (q, 2H),
2.04 (s, 3H), 2.16 (q, 2H), 2.72 (bs, 1H),
2.84 (s, 3H), 3.12 (m, 3H), 3.24 (s, 3H),
3.60 (dd, 1H, J=14, 2Hz), 4.00 (dd, 1H,
J = 12, 6Hz), 4.40 (d, 1H, J = 14Hz),
5.04 (t, 1H, J14Hz), 5.38 (dd, 1H, J=12,
6Hz), 7.48 (BS, 2H). Example 25 Compound 7-[(methoxyamino)methylene]amino-
9a-Methoxymitozane 660 mg of compound in 10 ml of methanol
(1.7mM) solution and add 170mg (2.0mM)
of methoxyamine and hydrochloric acid were added. The solution was stirred at 10° C. for 3 hours and at room temperature for 2 hours. TLC revealed only traces of unreacted compounds. The black precipitate formed upon standing was collected and washed with acetone to obtain 380 mg (57%) of the desired product. Elemental analysis: ( _C17H21N5O6 ) Calculated: C, _52.19 ; H, 5.40; N, 17.90 Measured _: C, 51.64; _H , 5.40; N, 17.83. UV (MeOH) λ _nax. nm: 376, 242. IR (KBr) ν _nax ′cm ^-1 : 3440, 3250, 3140, 2920,
1730, 1645, 1615, 1560, 1450, 1320, 1050. ^{The 1} H NMR (pyridine d ₅ ) spectrum was consistent with the structure of the title compound and the tautomeric form at C-7. Example 26 7-[(benzoyloxyamino)methylene]amino-9a-methoxymitozane A solution of 100 mg (0.26 mM) of the compound in 2 ml methanol containing 0.5 ml triethylamine was prepared and 400 mg (2.5 mM) o-benzyl-hydroxylamine hydrochloric acid was added. 2.5 Reactions at room temperature
Time progressed. tlc (CH ₂ Cl ₂ /methanol 10:
In 1), an orange/brown main component band appeared beyond the green band corresponding to the compound. The reaction mixture was concentrated to a residue and it was saturated with silica gel (20
g) Flash chromatographic analysis was carried out on top using CH ₂ Cl ₂ /methanol 20:1 as eluent.
The main brown band constituting the desired product was collected to give 80 mg (65.6% yield) of an amorphous solid. Elemental analysis: ( _C23H25N5O6 ) _Calculated : C, 59.10; _H , 5.35; N, 14.97 Measured _: C, 58.43; H, 5.48; N, 14.62. UV (MeOH) λ _nax゜nm: 376, 245, 209. IR (KBr) ν _nax ′cm ^-1 : 3460, 3300, 2945, 2920,
1745, 1720, 1570, 1275, 1220, 1060. ^{The 1} H NMR (pyridine d ₅ ) spectrum was consistent with the title compound and the C-7 tautomeric form [Formula]. 10 mg of unreacted starting material compound was recovered. Example 27 Compound 7-(1,3-dimethyl-2-imidazolinylidene)-9a-methoxymitozane Dissolve 0.34 g (1 mM) mitomycin C in 5 ml 1,3-dimethyl-2-imidazolidone and add 0.1 g sodium hydride (50% in oil,
2.08mM) was added. The mixture was kept at room temperature for 20 minutes and then cooled in an ice-salt bath (-15°C). After keeping the mixture at this temperature for 10 minutes, 0.65g (2mM)
of 2-chloro-1,3-dimethyl-4,5-dihydro-(3H)-imidazoliminium chloride was added. After being kept at -5°C for 1 hour, it was diluted with ethyl acetate and chromatographed on an alumina column. The column was eluted with methylene chloride, methylene chloride containing 2% v/v methanol. A green fraction corresponding to the desired product was obtained and an additional 10
It was purified by chromatography on alumina using methylene chloride containing % v/v methanol. Elemental analysis: ( _{C20H26N6O5.1-1} / _4H2O ) Calculated: C, _53.03 ; H, 6.34; _N , _18.55 Measured: C, 52.68; H, 6.21; N, 18.15. NMR (pyridine- _d5 , δ): 2.32 (s, 3H), 2.47
(s, 3H), 2.59 (s, 3H), 2.74 (m, 1H),
3.03−3.32 (m, 5H), 3.26 (s, 3H), 3.66 (bd,
1H, J=12Hz), 4.02 (dd, 1H, J=11, 4
Hz), 4.75 (d, 1H, J=12Hz), 5.09 (bt, 1H,
J = 11Hz), 5.44 (dd, 1H, J = 11, 4Hz). IR (KBr): 3400, 3280, 2930, 1700, 1610,
1480, 1330, 1055cm ^-1 . UV (MeOH, λ _nax ): 600, 375, 252 (sh),
222nm. Example 28 Compound 7-[(1,3-dimethyltetrahydropyrimidinylidene)amino-9a-methoxymitozane A solution of mitomycin C (680 mg, 2 mM) in 8 ml of 1,3-dimethyl-3,4,5,6-tetrahydro(1H,3H)-2-pyrimidinone was treated with sodium hydride (50% oil dispersion, 200mg,
4.2mM) was added. The mixture was kept at room temperature for 20 minutes and then cooled to -25°C. 2-chloro-1,3
-Dimethyl-2,3,4,5-tetrahydro-pyrimidinium chloride 0.73g (4mM) was added and the mixture was kept at -25°C for 3 hours. It was then diluted with ethyl acetate and 2 ml of methanol. The mixture was placed on a dry alumina chromatographic column and first diluted with methylene chloride and then 2% v/v.
Elution with methanol/methylene chloride gave 0.35 g (39.5% yield) of the desired product. Melting point 138-140
℃. Elemental analysis: ( _{C21H27N6O5.H2O} ) _Calculated value: C, _54.65 ; H, 6.33; N, _18.21 Measured value _: C, 54.78; H, 6.18; N, 18.21. NMR (pyridine- _d5 , δ): 1.80 (m, 2H), 2.42
(s, 3H), 2.52 (s, 3H), 2.64 (s, 3H),
2.76 (m, 1H), 2.90-3.60 (m, 5H), 3.26 (s,
3H), 3.74 (d, 1H, J=12Hz), 4.05 (dd,
1H, J=11, 4Hz), 4.97(d, 1H, J=12
Hz), 5.09 (t, 1H, J=11Hz), 5.41 (dd, 1H,
J=11, 4Hz). IR (KBr): 3430, 3280, 2930, 1710, 1570,
1480, 1450, ^{1350, 1050cm -1} . UV (MeOH, λ _nax ): 635, 377, 264 (sh),
223nm. Example 29 Compound 7-(tetramethyldiaminomethylene)amino-9a-methoxymitozane 425 mg (1.42 mM) of mitomycin C was mixed with 85.3 mg of sodium hydride in a 50% oil dispersion, to which 4 ml of dimethylformamide was added. The mixture was stirred for 10 minutes under an atmosphere of argon, then cooled to -35°C. Add 289mg (2.13mM) of tetramethylchloroformamidiminium chloride.
The mixture was warmed to 5° C. over time. Ground tri-ice was added to the mixture to quench the reaction, and then the solvent was removed by distillation under reduced pressure. The residue was chromatographed on an alumina column (100 g) using 3% v/v methanol in methylene chloride as elution. This material was further purified on alumina (5% v/v methanol in methylene chloride) and
Two fractions of 17 mg and 76 mg were obtained. The latter was recrystallized from acetone-ether to give the desired product. Melting point 193-195℃. (12% yield). Elemental analysis: ( _C20H28N6O5 ) Calculated: C, 55.54; H, 6.53; _N , 19.43 Measured _: C _, 54.92; H, 6.53; N, 19.29. NMR (pyridine- _d5 , δ): 2.26 (s, 3H), 2.59
(s, 6H), 2.68 (s, 6H), 2.75 (m, 1H),
3.15 (d, 1H, J=4Hz), 3.26 (s, 3H),
3.65 (d, 1H, J = 12Hz), 4.00 (dd, 1H, J =
11, 5Hz), 4.62 (d, 1H, J=12Hz), 5.04
(t, 1H, J=11Hz), 4.38 (dd, 1H, J=11,
5Hz). IR (KBr): 3430, 3280, 2920, 1710, 1610,
1495, 1335, 1055 cm ^-1 . UV (MeOH, λ _nax ): 610, 80, 260, 220nm. Example 30 Compound XI 7-(1-piperidinylmethylene)-amino-
9a-Methoxymitozane Oxalyl chloride (380mg, 3mM) was added dropwise to 6ml of chloroform containing 0.34g (3mM) of 1-formylpiperidine to prepare 0.5M of piperidinylchloromethyleniminium chloride.
A solution was prepared. Separately, a solution of mitomycin C (334 mg, 1 mM) in 3 ml of 1-formylpiperidine was added under nitrogen under sodium hydrogen (50% oil dispersion, 96%
mg, 2mM) was added. After stirring for 15 minutes at room temperature, the solution was cooled to -25°C and the iminium salt solution prepared above (4ml, 2mM) was added. 1 of the reaction mixture
The reaction was kept at −25° C. for an hour and then added with dry ice to quench the reaction. After adding methanol (1 ml), the product mixture was adsorbed onto neutral alumina. This material was placed on an alumina column (30 g) and the column was eluted first with methylene chloride and then with 3% v/v methanol in methylene chloride to give 360 mg.
(84%) of the title compound was obtained. Melting point 68-70℃. Elemental analysis: ( _{C21H25N5O6.1-1 / 4H2O} ) Calculated: C, _55.80 ; H, 6.58; N, _15.49 Measured: C, 55.57; H, 6.21; N, 15.91. NMR (pyridine- _d5 , δ): 1.42 (bs, 6H), 2.19
(s, 3H), 2.72 (m, 1H), 3.06−3.30 (m,
3H), 3.25 (s, 3H), 3.48−3.70 (m, 2H),
3.57 (d, 1H, J = 13Hz), 4.01 (dd, 1H, J =
11, 4Hz), 4.43 (d, 1H, J=13Hz), 5.02
(bt, 1H, 3=11Hz), 5.55 (dd, 1H, J=11,
4Hz), 7.86 (s, 1H). IR (KBr): 3440, 3350, 3300, 2935, 2835,
1710, 1615, 1520, 1445, 1305, 1250, 1200,
1055cm _- ¹ . UV (MeOH, λ _nax ): 590, 389, 262 (sh), 234,
212(sh)nm. Example 31 7-Hydroxy- ^N10 -dimethylaminomethylene-9a-methoxymitozane To a solution of 7-hydroxy-9a-methoxymitozane (20 mg) in methylene chloride (3 ml) was added dimethylformamide dimethyl acetal (1 ml) and the solution was stirred for 30 minutes at about 65°C. The progress of the reaction was followed by tic (10:1 CH ₂ Cl ₂ /MeOH). The product was collected by concentrating the mixture under reduced pressure and the residue was chromatographed on silica gel to give the title compound. Activity against P-388 Murine Leukemia Experiments in which CDF ₁ female mice implanted intraperitoneally with a tumor inoculum of ¹⁰⁶ ascites cells of P-388 Murine Leukemia were treated with various doses of the test compound of formula or mitomycin C. The results of the laboratory test are shown in Table 4.
Compounds were administered by intraperitoneal injection. Groups of 6 mice were used for each dose level and were given a single dose of compound only once a day. Ten
One saline-treated control mouse was included in each experimental series. A group receiving mitomycin C was included as a positive control. Measured in days for each group of mice. A 30-day protocol was adopted for mean survival time. Also
The number of survivors at the end of the 30 day period was recorded. Mice were weighed before treatment and on day 6. Changes in body weight were used as a measure of drug toxicity. With mice weighing 20 g each, weight loss of up to approximately 2 g was not considered excessive. The results are the mean survival time of the control group given saline (100)
It was measured using %T/C, which is the ratio of the mean survival time of the group administered the compound to that of the group administered the compound. Saline-treated control animals usually died within 9 days. The maximum effect in the following table is shown by the %T/C and dose at which the effect was achieved. The numbers in brackets are the values when mitomycin C was used as a positive control in the same experiment. Therefore, the relative activity of the substance of the present invention against mitomycin C could be measured.
The minimum effect in %T/C should be considered to be about 125. The dose that gave a %T/C of approximately 125 was reported as the minimum effect in the table below. The two values in the mean weight change column are the weight change per individual mouse at the maximum and minimum effective doses. [Table] [Table] Compounds and are of particular interest. This is because its activity clearly exceeds mitomycin C, both in terms of maximum efficacy and milligram efficacy (dose comparison for equal efficacy). A is the amidino group;
A compound of formula (), wherein B is _-NH2 . In other words, it is a mitomycin C derivative in which R ₂ , R ³ and R ⁴ are as defined above, and N ⁷ is substituted with an aminomethylene group of the formula [Formula]. The bis-amidino compounds of the present invention, wherein each of A and B is said amidino group, are also of substantial interest as active antitumor agents. See data for compounds meeting this structural requirement in Table 4. Table 5 shows the results of an antitumor test using B16 melanocytoma produced in mice. BDF ₁ mice were used and inoculated intraperitoneally with tumor implants. A 60 day protocol was used. 10 for each dose tested
Groups of 1 mouse were used to determine mean survival time. Control animals were inoculated and treated with injection vehicle in the same manner as the test animals, but none of the drugs showed a mean survival time of 21 days. The respective survival time (%T/C) relative to that of the control was used as a measure of efficacy. The highest and lowest effective doses were determined for each compound tested. The minimum effective dose is
It was defined as the dose that showed a %T/C value of 125. For each dose level, test animals were treated with the test compound by intraperitoneal method on days 1, 5, and 9. The change in body weight per day at the highest and lowest effective doses was used as a measure of toxicity. A weight loss of 2 g for a 20 g mouse was not excessive. [Table] [Table] Compound (Example 29) and Compound (Example 28) were effective against B16 murine melanocytoma.
A subcutaneous method of tumor inoculation was used and tested by intravenous drug administration. The treatment plan and survival time evaluation (adopting a 40-day protocol) were determined in the same manner as in Table 5. Changes in body weight after 12 days were measured. The highest effective dose of the compound was 156% T/C1
mg/Kg, and the body weight increased by 1.5g. Groups of 6 animals were used, and at this dose 3 animals survived the 40 day protocol. The minimum effective dose was 0.25 mg/Kg. At this dose, the change in body weight after 12 days was 1.0 g. The highest effective dose for the compound is 8 for a %T/C of 177.
mg/Kg, and the weight change was -0.6.
The minimally effective dose was 4 mg/Kg with body weight change +0.8. The most effective dose of mitomycin C in the same experiment was 3 mg/Kg with a % T/C of 195 and a body weight change of -0.5. The minimum effective dose of mitomycin C was not determined. In a short toxicological protocol using groups of 5 male BDF ₁ mice per dose administered intraperitoneally alone, the compound was administered at the most desirable effective dose (1.6 mg/Kg ip). ), no significant decrease occurred in lymphocyte count. The lack of significant increases in blood urea nitrogen (BUN) or serum glutamine phosphotransferase (SGPT) at this dose may be due to the absence of adverse effects of suppression of renal or hepatic function or lymphatic activity. It is shown that. In view of the outstanding antitumor activity observed in experimental animal tumors and the low toxicity compared to mitomycin C, the present invention encompasses the use of substances of formula () for the inhibition of mammalian tumors. To this end, substantially non-toxic, anti-tumor effective doses are systematically administered to the tumor-bearing mammal.

Claims (1)

[Claims] 1 General formula () [In the formula, A is amino, methoxy, hydroxy,
(1-lower alkyl-2(1H)-pyridinylidene)
Amino or formula [formula] [formula] [formula] [formula] [formula] [formula] B is amino or formula is an amidino group, and at least one of A or B is one of the specified groups other than amino, methoxy and hydroxy, n is an integer of 0, 1, 2, or 3, and R ¹ is hydrogen , lower alkyl or lower alkanoyl, R ² is hydrogen or lower alkyl, R ³ is lower alkyl, lower alkoxy, or R ⁴ and the nitrogen to which both (R ³ , R ⁴ ) are bonded. With atoms: pyrrolidine, 2- or 3-lower alkylpyrrolidine, piperidine, 2-, 3
-, or 4-lower alkylpiperidine, 2,6-
di-lower alkylpiperidine, piperazine, 4-substituted piperazine (wherein the 4-substituents are alkyl or carbalkoxy each having 1 to 8 carbon atoms; phenyl, methylphenyl, methoxyphenyl, halofenyl, nitrophenyl, or benzyl) , azepine, 2-, 3-, 4-, or 5-lower alkyl azepine, morpholine, thiomorpholine, thiomorpholine-1-oxide or thiomorpholine-1,1-dioxide, and R ⁴ is lower alkyl. or R ³ and the nitrogen atom to which they are bonded together with pyrrolidine, 2- or 3-lower alkylpyrrolidine, piperidine, 2
-, 3-, or 4-lower alkylpiperidine,
2,6-dilower alkylpiperidine, piperazine, 4-substituted piperazine (however, each of the 4 substituents is 1
alkyl or carbalkoxy having from 8 carbon atoms; phenyl, methylphenyl, methoxyphenyl, halofenyl, nitrophenyl, or benzyl), azepine, 2-, 3-, 4-,
or form a 5-lower alkyl azepine, morpholine, thiomorpholine, thiomorpholine-1-oxide or thiomorpholine-1,1-dioxide, ^R5 is _C1-18 alkyl other than tertiary alkyl, ^R7 and R ⁹ is independently H or lower alkyl, and the aforementioned groups such as lower alkyl, lower alkanoyl and lower alkoxy consist of 1 to 6 carbon atoms. 2 A and B each independently have the formula The compound according to claim 1, which is the amidino group of 3. The compound according to claim 2, wherein R ¹ and R ² are each hydrogen, and R ³ and R ⁴ are each methyl. 4 7-[(dimethylamino)methylene]amino-
The compound according to claim 1, which is N ¹⁰ -(dimethylamino)methylene-9a-methoxymitozane. 5 7-[(dimethylamino)methylene]amino-
The compound according to claim 1, which is ^N10- (dimethylamino)methylene- ^N1a -formyl-9a-methoxymitozane. 6. The compound according to claim 1, which is 7-[(diisopropylamino)methylene]amino- ^N10- (diisopropylamino)methylene-9a-methoxymitozane. 7 7-[(dimethylamino)methylene]amino-
The compound according to claim 1, which is ^N10- (dimethylamino)methylene-9a-methoxy- ^N1a -methylmitozane. 8 R ¹ and R ² are each hydrogen, and R ³ and
3. The compound according to claim 2, wherein R ⁴ forms a piperidine group together with the nitrogen atom to which they are bonded. 9 N ^1a -formyl-9a-methoxy-7-(1-
The compound according to claim 1, which is (piperidinylmethylene)amino- ^N10- (1-piperidinylmethylene)mitozane. The compound according to claim 1, which is 109a-methoxy-7-(1-piperidinylmethylene)amino- ^N10- (1-piperidinylmethylene)mitozane. 11. The compound according to claim 2, wherein R ¹ and R ² are each hydrogen, and R ³ and R ⁴ together with the nitrogen atom to which they are bonded form a morpholine group. 12. The compound according to claim 1, which is 129a-methoxy-7-(1-morpholino-methylene)-amino- ^N10- (1-morpholino-methylene)mitozane. 13 A is an amino group and B is the formula The compound according to claim 1, which is an amidino group of 14. The compound according to claim 13, wherein R ³ and R ⁴ are each methyl. 15. The compound according to claim 1, which is 7-amino- ^N10 -dimethylaminomethylene-9a-methoxy- ^N1a -methylmitozane. 16. The compound according to claim 1, which is 7-amino- ^N10 -dimethylaminomethylene-9a-methoxymitozane. 17 A is a methoxy group and B is the formula The compound according to claim 1, which is an amidino group of 18. The compound according to claim 17, wherein R ³ and R ⁴ are each methyl. 19 Claim 1 which is 7,9a-dimethoxy-N ¹⁰ -dimethylaminomethylene mitozane
Compounds described in Section. 20 7,9a-dimethoxy-N ¹⁰ -dimethylaminomethylene-N ^1a -formylmitozane. 21. The compound according to claim 1, wherein A is one of the amidino groups and B is an amino group. 22 A is the formula The compound according to claim 21, wherein the compound is an amidino group, and R ³ and R ⁴ are methyl groups. 23 7-(dimethylaminomethylene)amino-
The compound according to claim 1, which is 9a-methoxymitozane. 24 7-(dimethylaminomethylene)amino-
The compound according to claim 1, which is 9a-methoxy-N ^1a -methyl-mitozane. 25 7-(1-methyl-2(H)-pyridinylidene)
The compound according to claim 1, which is amino-9a-methoxymitozane. 26. The compound according to claim 1, which is 9a-methoxy-7-(1-morpholinomethylene)aminomitozane. 27 The compound according to claim 1, which is 7-(1-pyrrolidinylmethylene)amino-9a-methoxymitozane. 28 7-(1-dimethylamino)ethylidene]
The compound according to claim 1, which is amino- ^N10- [1-dimethylamino)ethylidene]-9a-methoxymitozane. 29 The compound according to claim 1, which is 7-[1-(dimethylamino)ethylideneamino]-9a-methoxymitozane. 30 7-[(1-methyl-2-pyrrolidinidene)
The compound according to claim 1, which is amino]-N ¹⁰ -[(1-methyl-2-pyrrolidinylidene)amino]-9a-methoxymitozane. 31 The compound according to claim 1, which is 7-[(1-methyl-2-pyrrolidinylidene)amino]-9a-methoxymitozane. 32 The compound according to claim 1, which is 7-[(methoxyamino)methylene]amino-9a-methoxymitozane. 33 7-[(benzyloxyamino)methylene]
The compound according to claim 1, which is amino-9a-methoxymitozane. 34 The compound according to claim 1, which is 7-(1,3-dimethyl-2-imidazolidylidene)-9a-methoxymitozane. The compound according to claim 1, which is 35 7-[(1,3-dimethyltetrahydropyrimidinylidene)-amino-9a-methoxymitozane. 36 The compound according to claim 1, which is 7-(tetramethyldiaminomethylene)amino-9a-methoxymitozane. 37 The compound according to claim 1, which is 7-(1-piperidinylmethylene)amino-9a-methoxymitozane. 38 7-[2-benzylthioethyl]amino-
9a-Methoxymitozane. 39 A compound selected from the group consisting of mitomycin C, mitomycin A, or N ^1a -lower alkyl derivatives of both of the above, and the general formula in a solution. [In the formula, R ² is hydrogen or lower alkyl, R ³ is lower alkyl, lower alkoxy, or
Or, together with R ⁴ and the nitrogen atom to which both (R ³ , R ⁴ ) are bonded, pyrrolidine, 2- or 3-lower alkylpyrrolidine, piperidine, 2-, 3
-, or 4-lower alkylpiperidine, 2,6-
di-lower alkylpiperidine, piperazine, 4-substituted piperazine (wherein the 4-substituents are alkyl or carbalkoxy each having 1 to 8 carbon atoms; phenyl, methylphenyl, methoxyphenyl, halofenyl, nitrophenyl, or benzyl) , azepine, 2-, 3-, 4-, or 5-lower alkyl azepine, morpholine, thiomorpholine, thiomorpholine-1-oxide or thiomorpholine-1,1-dioxide, and R ⁴ is lower alkyl. or R ³ and the nitrogen atom to which they are bonded together with pyrrolidine, 2- or 3-lower alkylpyrrolidine, piperidine, 2
-, 3-, or 4-lower alkylpiperidine,
2,6-dilower alkylpiperidine, piperazine, 4-substituted piperazine (however, each of the 4 substituents is 1
alkyl or carbalkoxy having from 8 carbon atoms; phenyl, methylphenyl, methoxyphenyl, halofenyl, nitrophenyl, or benzyl), azepine, 2-, 3-, 4-,
or to form a 5-lower alkyl azepine, morpholine, thiomorpholine, thiomorpholine-1-oxide or thiomorpholine-1,1-dioxide, each R ⁸ being independently lower alkyl having up to 6 carbon atoms or cycloalkyl or taken together to form alkylene, together with the oxygen atom to which it is attached and any intervening carbon atoms, 5 or 6
with an amide acetal of the general formula () at 40 to 65 °C in a liquid organic reaction medium compatible with anhydrous reaction. Both A and B of The general formula () is reacted until a reaction product, which is an amidino group of is an amidino group, R ¹ is hydrogen, lower alkyl or lower alkanoyl, R ² , R ³ and R ⁴ are as described above; A method for producing a compound having 6 carbon atoms. 40 The liquid organic reaction medium is a halogenated lower aliphatic hydrocarbon, and the amide acetal is used in a ratio of 2 moles or more to mitomycin C to produce a product in which both A and B are the amidino groups. 39. The method according to claim 39. 41. The method of claim 40, wherein the reaction medium is chloroform. 42. The method of claim 40, wherein the reaction medium is a mixture of a halogenated lower aliphatic hydrocarbon and a lower alkanol. 43. The method of claim 40, wherein the reaction medium is a mixture of chloroform and methanol. 44 General formula [In the formula, each of A and B is the formula is an amidino group, R ¹ is hydrogen, lower alkyl or lower alkanoyl, R ² is hydrogen or lower alkyl, R ³ is lower alkyl, lower alkoxy, or R ⁴ and both (R ³ , R ⁴ ) together with the nitrogen atom to which pyrrolidine, 2- or 3-lower alkylpyrrolidine, piperidine, 2-, 3
-, or 4-lower alkylpiperidine, 2,6-
di-lower alkylpiperidine, piperazine, 4-substituted piperazine (wherein the 4-substituents are alkyl or carbalkoxy each having 1 to 8 carbon atoms; phenyl, methylphenyl, methoxyphenyl, halofenyl, nitrophenyl, or benzyl) , azepine, 2-, 3-, 4-, or 5-lower alkyl azepine, morpholine, thiomorpholine, thiomorpholine-1-oxide or thiomorpholine-1,1-dioxide, and R ⁴ is lower alkyl. or R ³ and the nitrogen atom to which they are bonded together with pyrrolidine, 2- or 3-lower alkylpyrrolidine, piperidine, 2
-, 3-, or 4-lower alkylpiperidine,
2,6-dilower alkylpiperidine, piperazine, 4-substituted piperazine (however, each of the 4 substituents is 1
alkyl or carbalkoxy having from 8 carbon atoms; phenyl, methylphenyl, methoxyphenyl, halofenyl, nitrophenyl, or benzyl), azepine, 2-, 3-, 4-,
or 5-lower alkyl azepine, morpholine, thiomorpholine, thiomorpholine-1-oxide or thiomorpholine-1,1-dioxide is formed, and the aforementioned groups such as lower alkyl, lower alkanoyl and lower alkoxy contain 1 to 6 consisting of carbon atoms] with an aliphatic, cycloaliphatic, aromatic, heteroaromatic or heteroalicyclic primary amine in a liquid organic reaction medium compatible with anhydrous reaction at about -15°C. A method for producing 7-amidino-9a-methoxymitozane, which comprises reacting at a temperature of 50°C to 50°C. 45. The process of claim 44, wherein the liquid organic reaction medium compatible with anhydrous reactions consists of methanol, chloroform, methylene chloride, or other lower halogenated alkanes. 46 General formula () [In the formula, each of A and B is the formula is an amidino group, R ¹ is hydrogen, lower alkyl or lower alkanoyl, R ² is hydrogen or lower alkyl, R ³ is lower alkyl, lower alkoxy, or R ⁴ and both (R ³ , R ⁴ ) together with the nitrogen atom to which pyrrolidine, 2- or 3-lower alkylpyrrolidine, piperidine, 2-, 3
-, or 4-lower alkylpiperidine, 2,6-
di-lower alkylpiperidine, piperazine, 4-substituted piperazine (wherein the 4-substituents are alkyl or carbalkoxy each having 1 to 8 carbon atoms; phenyl, methylphenyl, methoxyphenyl, halofenyl, nitrophenyl, or benzyl) , azepine, 2-, 3-, 4-, or 5-lower alkyl azepine, morpholine, thiomorpholine, thiomorpholine-1-oxide or thiomorpholine-1,1-dioxide, and R ⁴ is lower alkyl. or R ³ and the nitrogen atom to which they are bonded together with pyrrolidine, 2- or 3-lower alkylpyrrolidine, piperidine, 2
-, 3-, or 4-lower alkylpiperidine,
2,6-dilower alkylpiperidine, piperazine, 4-substituted piperazine (however, each of the 4 substituents is 1
alkyl or carbalkoxy having from 8 carbon atoms; phenyl, methylphenyl, methoxyphenyl, halofenyl, nitrophenyl, or benzyl), azepine, 2-, 3-, 4-,
or 5-lower alkyl azepine, morpholine, thiomorpholine, thiomorpholine-1-oxide or thiomorpholine-1,1-dioxide is formed, and the aforementioned groups such as lower alkyl, lower alkanoyl and lower alkoxy contain 1 to 6 consisting of carbon atoms] at 20°C to 60°C with an amine selected from the group consisting of aminodiphenylmethane, trifluoroethylamine, and tertiary butylamine in a molar proportion or more. and B is an amino group. 47. The method of claim 46, wherein the reaction is carried out in an organic reaction medium compatible with anhydrous reactions with methanol, chloroform, methylene chloride, or other halogenated alkanes. 48 Reacting a solution of mitomycin C in dimethylformamide (or other suitable solvent) with 0.1 to 1.5 molar proportions of sodium hydride to form an anionic form of mitomycin C and converting the anionic form into an iminoether, iminothioether, etc. , selected from the group consisting of halomethyleniminium halides and iminohalide salts, of the formula [However, R ² is hydrogen or lower alkyl, R ³ is lower alkyl or lower alkoxy, or together with R ⁴ and the nitrogen atom to which both (R ³ , R ⁴ ) are bonded, pyrrolidine, 2 -, or 3-lower alkylpyrrolidine, piperidine, 2-, 3
-, or 4-lower alkylpiperidine, 2,6-
di-lower alkylpiperidine, piperazine, 4-substituted piperazine (wherein the 4-substituent is alkyl or carbalkoxy each having 1 to 8 carbon atoms; phenyl, methylphenyl, methoxyphenyl, halofenyl, nitrophenyl, or benzyl) , azepine, 2-, 3-, 4-, or 5-lower alkyl azepine, morpholine, thiomorpholine, thiomorpholine-1-oxide or thiomorpholine-1,1-dioxide, and R ⁴ is lower alkyl. or R ³ and the nitrogen atom to which they are bonded together with pyrrolidine, 2- or 3-lower alkylpyrrolidine, piperidine, 2
-, 3-, or 4-lower alkylpiperidine,
2,6-dilower alkylpiperidine, piperazine, 4-substituted piperazine (provided that each substituent is alkyl or carbalkoxy having 1 to 8 carbon atoms; phenyl, methylphenyl, methoxyphenyl, halofenyl, nitrophenyl, or benzyl) ), azepine, 2-, 3-, 4-,
or 5-lower alkylazepine, morpholine, thiomorpholine, thiomorpholine-1-oxide or thiomorpholine-1,1-dioxide] with an electrophilic reagent capable of forming the amidino group. () [However, A is the formula is a group represented by, B is _-NH2 , n is an integer of 0, 1, 2, or 3, ^R1 is hydrogen, lower alkyl, or lower alkanoyl, ^R2 , ^R3, and ^R4 is as described above, and the aforementioned lower alkyl, lower alkanoyl and lower alkoxy groups consist of 1 to 6 carbon atoms. 49 General formula [In the formula, A is amino, methoxy, hydroxy,
(1-lower alkyl-2(1H)-pyridinylidene)
Amino or formula [formula] [formula] [formula] [formula] [formula] [formula] is a group represented by, B is amino or a group represented by the formula is an amidino group, and at least one of A or B is one of the specified groups other than amino, methoxy and hydroxy, n is an integer of 0, 1, 2, or 3, and R ¹ is hydrogen , lower alkyl or lower alkanoyl, R ² is hydrogen or lower alkyl, R ³ is lower alkyl, lower alkoxy, or R ⁴ and the nitrogen to which both (R ³ , R ⁴ ) are bonded. With atoms: pyrrolidine, 2- or 3-lower alkylpyrrolidine, piperidine, 2-, 3
-, or 4-lower alkylpiperidine, 2,6-
di-lower alkylpiperidine, piperazine, 4-substituted piperazine (wherein the 4-substituents are alkyl or carbalkoxy each having 1 to 8 carbon atoms; phenyl, methylphenyl, methoxyphenyl, halofenyl, nitrophenyl, or benzyl) , azepine, 2-, 3-, 4-, or 5-lower alkyl azepine, morpholine, thiomorpholine, thiomorpholine-1-oxide or thiomorpholine-1,1-dioxide, and R ⁴ is lower alkyl. or R ³ and the nitrogen atom to which they are bonded together with pyrrolidine, 2- or 3-lower alkylpyrrolidine, piperidine, 2
-, 3-, or 4-lower alkylpiperidine,
2,6-dilower alkylpiperidine, piperazine, 4-substituted piperazine (however, each of the 4 substituents is 1
alkyl or carbalkoxy having from 8 carbon atoms; phenyl, methylphenyl, methoxyphenyl, halofenyl, nitrophenyl, or benzyl), azepine, 2-, 3-, 4-,
or 5-lower alkyl azepine, morpholine, thiomorpholine, thiomorpholine-1-oxide or thiomorpholine-1,1-dioxide is formed, R ⁵ is C _1-18 alkyl other than tertiary alkyl, and R ⁷ and R ⁹ is independently H or lower alkyl, and the aforementioned lower alkyl, lower alkanoyl, lower alkoxy, etc. groups consist of 1 to 6 carbon atoms.