BE896963A - AMIDINES - Google Patents

Abstract

Compound of formula (I), where A represents an amino, methoxy, hydroxyl, (1-lower alkyl -2 (1H) -pyridinylidene) amino radical or a radical of formula (II), B represents an amino or amidino radical and minus one of A and B represents one of the specified radicals other than amino, methoxy or hydroxyl, n represents 0,1,2 or 3. The amidines derived from mitomycin C are useful intermediates for reactions with primary amines giving 7-amino-9a-methoxymitosans, some of which are known and others which are new and which show activity against certain experimental tumors in animals.

Description

       

  Amidines

  
The present invention relates to mitomycin analogs which contain one or more amidino radicals (class 260, subclass 326.24). These compounds are derivatives of mitomycin C in which the 7-amino radical and / or the nitrogen atom of the carbamido radical are part of an amidino radical. These compounds exhibit activity against experimental tumors in animals.

  
According to the systematic nomenclature of Chemical Abstracts, mitomycin C is called the

  
  <EMI ID = 1.1>

  
number of the azirinopyrroloindole ring system being as follows:

  

  <EMI ID = 2.1>

Chemical Abstracts

  
According to current terminology in the field of mitomycins, this ring system carrying different substituents characteristic of mitomycins is called mitosan.

  
&#65533;

  

  <EMI ID = 3.1>


  
Mitosan

  
This nomenclature is suitable for simple derivatives such as those bearing substituents on the nitrogen atom of the azirino radical and at positions 7 and 9a, but exposes to some ambiguities and drawbacks. No traditional numbering system distinguishes between the nitrogen atom of the azirino radical and the nitrogen atom

  
of the carbamoyl radical in the side chain which carry both of the substituents in certain of the compounds of the invention. For this reason, the nitrogen atom <EMI ID = 4.1>

  
  <EMI ID = 5.1>

  
mitosanes for the purposes of the invention. When the compounds of the invention are identified by the term "mitosan" or by the structural formula, their stereochemical configuration is that of mitomycin C.

  
Mitomycin C is an antibiotic produced by fermentation and currently used in particular in the United States of America for the treatment of concealed adenocarcinoma of the stomach or pancreas in proven combinations with other recognized chemotherapeutic agents, as well as for palliative treatment after the failure of another therapy

  
  <EMI ID = 6.1>

  
13201, Physicians' Desk Reference 35th edition, 1981, pages 717 and 718). Mitomycin C and its preparation by fermentation are the subject of United States patent n [deg.] 3,660,578 based in particular on a patent application in Japan of April 6, 1957.

  
The structures of mitomycins A, B and C and porfiromycin have been published by J.S. Webb et al. from Lederle Laboratories Division American Cyanamid Company, J. Amer. Chem. Soc. 84, 3185-3187
(1962). One of the chemical transformations carried out during this structural study to establish the relationship between mitomycin A and mitomycin C is the conversion of the first or 7-9 d-dimethoxymitosan, by reaction with ammonia in the second or 7-amino9o (-methoxymitosan. Elimination of the 7-methoxy radical from mitomycin A has proved to be a reaction of considerable interest for preparing the antitumor derivatives of mitomycin C. all relate to the conversion of mitomycin A to an antitumor 7- (substituted amino) -mitomycin C.

   This research aimed to prepare more active derivatives, but less toxic than mitomycin C: Matsui et al. "The Journal of Antibiotics", XXI, 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, and Bradner, Abstracts of Papers Annual Meeting of the American Chemical Society, Las Vegas, Nevada, March 1982,

  
Abstract No. MEDI 72.

  
The following patents relate to the preparation of 7- (substituted amino) -mitosans by reaction of mitomycin A, mitomycin B or one of their N derivatives -substituted with a primary or secondary amine: Cosulich et al. EUA patent No. 3,332,944 (July 25, 1967); Matsui et al. EU Patent No. 3,420,846 (January 7, 1969); Matsui et al. EU Patent No. 3,450,705 (June 17, 1969); Matsui et al. EUA patent No 3,514,452 (May 26
1970); Nakano et al. EUA Patent No. 4,231,936 (November 4
1980); Remers EUA patent No 4,268,676 (May 19, 1981).

  
Different derivatives of mitomycin C carrying an amino radical substituted in position 7 have also been obtained by direct biosynthesis, in this case

  
by adding different primary amines to the fermentation broth and by conducting the fermentation of

  
the usual way to produce mitomycin

  
(C.A. Claridge et al. Abst. Of the Annual Metting

  
of Amer. Soc. for Microbiology 1982. Abs. 028).

  
Mitomycin C is the main mitomycin obtained by fermentation and constitutes the marketed form. The usual technique of converting mitomycin C to mitomycin A suitable for producing

  
semi-synthetic mitomycin C analogs

  
which carry a substituted amino radical, such as

  
is exposed in the aforementioned memoirs, consists in hydrolyzing mitomycin C into the corresponding 7-hydroxymitosan, which is a highly unstable compound, then in methylating the latter using diazomethane, which is a very dangerous substance to handle. To avoid having to methylate with diazomethane 7-0-demethylmitomycin A obtained by hydrolysis of mitomycin C,

  
we also tried to use 7-acyloxymitosans
(Kyowa Hakko Kogyo KK Japanese patent No. J5 6073-085, Farmdoc No. 56227 D / 31).

  
The present invention relates to a new group of monoguanidino- or mono- and bis-amidino-analogs of mitomycin C in which the nitrogen atom of the 7-amino radical and / or the nitrogen atom of the radical

  
  <EMI ID = 7.1>

  
cal amidino or the nitrogen atom of the 7-amino radical is part of a guanidino radical. Corresponding mitomycin A analogs carrying a methoxy radical

  
  <EMI ID = 8.1>

  
are also the subject of the invention. The compounds of the invention correspond to the following structural formula:

  

  <EMI ID = 9.1>


  
where A represents an amino, methoxy, hydroxyl radical,

  
  <EMI ID = 10.1>

  
radical of formula:

  

  <EMI ID = 11.1>
 

  
B represents an amino radical or the amidino radical

  
  <EMI ID = 12.1>

  
of formula R R N-C = N- and at least one of A and B represents one of the specified radicals other than amino, methoxy or hydroxyl,

  
n represents 0, 1, 2 or 3,

  
  <EMI ID = 13.1>

  
lower, lower alkanoyl, benzoyl or substituted benzoyl whose substituent is a lower alkyl, lower alkoxy, halo, amino or nitro radical,

  
R2 represents a hydrogen atom or a lower alkyl, phenyl, (lower alkyl) phenyl radical,
(lower alkoxy) phenyl, halophenyl, aminophenyl, nitrophenyl, thienyl, furyl, cyano, di (lower alkyl) amino, lower alkoxy or lower alkylthio,

  
R <3> represents a lower alkyl radical, lower alkoxy or constitutes with R4 and the nitrogen atom to which they are united a pyrrolidine radical, 2-, or 3-

  
  <EMI ID = 14.1>

  
(lower alkyl) piperidine, 2,6-di (lower alkyl) piperidine, piperazine, 4-substituted piperazine (of which

  
the substituent is an alkyl or carbalkoxy radical each having 1 to 8 carbon atoms, phenyl, methylphenyl, methoxyphenyl, halophenyl, nitrophenyl

  
or benzyl), azepine, 2-, 3-, 4- or 5- (lower alkyl) azepine, morpholine, thiomorpholine, thiomorpholine-1oxide or thiomorpholine-1,1-dioxide,

  
R4 represents a lower alkyl radical or constitutes with R <3> and the nitrogen atom to which they are united

  
a pyrrolidine radical, 2- or 3- (lower alkyl) pyrrolidine, piperidine, 2-, 3- or 4 - inférieur lower alkyl) piperidine, 2,6-di (lower alkyl) piperidine, piperazine, piperazine 4- substituted (including the substituent

  
is an alkyl or carbalkoxy radical, each of which has 1 to 8 carbon atoms, phenyl, methylphenyl, methoxyphenyl, halophenyl, nitrophenyl or benzyl), azepine, 2-, 3-, 4- or 5- (lower alkyl) azepine, morpholine, thiomorpholine, thiomorpholine-1-oxide or thiomorpholine-1,1-dioxide,

  
  <EMI ID = 15.1>

  
genoalkyl, Cl-18 hydroxyalkyl, C4-8 cycloalkyl, aryl or lower aralkyl each of which has up to 12 carbon atoms or a 3 to 8 membered heteroalicyclic or heteroaromatic radical of which at least two are carbon atoms ,

  
R 7 and R 9 independently represent hydrogen atoms or lower alkyl radicals,

  
it being understood that each of the lower alkyl, lower alkanoyl and lower alkoxy radicals mentioned above has 1 to 6 carbon atoms.

  
It is obvious to the specialist that some of the amidino radicals give rise to tautomeric forms. These are also defined by the formulas above and are the subject of the invention.

  
The substances of formula I above have anti-tumor activity in experimental animals. They are also useful as intermediates for the preparation of other compounds exhibiting this activity. The invention also relates to the methods for

  
to prepare the above substances and to transform them into other useful compounds having anti-tumor activity in experimental animals, as described below.

  
The process of the present invention using the above substances as intermediates

  
for the preparation of other compounds having an antitumor activity in animals comprises the reaction of a compound of formula I where A and / or B represent the indicated radiocalamidino with a primary amine leading

  
  <EMI ID = 16.1>

  
  <EMI ID = 17.1>

  
mitomycin A and mitomycin C. With a few exceptions, primary amines also react

  
at position 7 by eliminating the amidino radical and replacing it with the amino radical which they provide. These processes are illustrated by the following equations:

  

  <EMI ID = 18.1>


  
formula IV

  
R representing_the substituent carried by the nitrogen atom in various known and new 7- (amino-substituted) mitomycins C described here and primary amines capable of displacing the 7-amidino radical of formula II is chosen from the radicals Cl-18 alkyl

  
  <EMI ID = 19.1>

  
alkyl, C4-8 cycloalkyl, aryl, lower aralkyl or lower aralkoxy each of which has up to 12 carbon atoms and heteroalicyclic and heteroaromatic radicals having 3 to 8 members, at least two of which are carbon atoms. R representing the substituent carried by the nitrogen of the primary amines ca

  
  <EMI ID = 20.1>

  
is the remainder of a very weakly basic aliphatic amine or of a highly sterically hindered alkylamine or aralkoylamine. Examples are trifluoroethylamine, benzhydrylamine (or aminodiphenylmethane) and t-butylamine.

  
The compounds of formula I are obtained by reacting mitomycin C, 7-hydroxy-9a-methoxymitosan or mitomycin A or else a substituted N analog of any of these compounds with an amide-acetal. The compounds of formula I in which

  
A but not B represents this amidino radical can also be prepared by reaction of mitomycin C or one of its analogues N-substituted with a strong base

  
  <EMI ID = 21.1>

  
reaction of the anion with a reagent capable of generating the aminomethylene radical, such as a halogenomethyleniminium salt.

  
The preferred compounds which are the subject of the invention are the analogs of mitomycin C in which the 7-amino radical is part of a substituted or unsubstituted amidino radical. They show potent activity against experimental tumors in animals. These compounds are prepared by reacting mitomycin C with a reagent capable of transforming the 7-amino radical into a 7-amidino radical. The preferred reagents for this purpose are amide acetals which react in good yield and under moderate conditions with mitomycin C (Examples 1-5 and 18).

   Other amidine-forming reagents are imidoyl halides (example 17), halogenethyleneimium salts (example 15), 2-halogenoalkylpyridinium halides (example 16) and iminoethers or iminothioethers (examples 13 and 14) which react with the anionic form of mitomycin C resulting from the deprotonation of the 7-amino radical by reaction with a strong base. The conditions for the deprotonation of

  
mitomycin C amounts to reacting mitomycin C in solution in dimethylformamide with approximately 1.5 molar proportion of sodium hydride at room temperature. The reaction of the resulting anionic form with one of the abovementioned reagents is preferably carried out with 1 to 1.5 molar proportion thereof relative to mitomycin C between room temperature and around -60 [deg.] C. Polar aprotic organic solvents such as dimethylformamide, hexamethylphosphoramide, dimethylsulfoxide or pyridine

  
are used as reaction media. The method, however, is not limited to the formation of anionic mitomycin C in this way alone, since different variants are obvious to the specialist.

  
The preferred method for preparing the compounds of formula I where B or A and 2B represent the

  
  <EMI ID = 22.1>

  
amidino radical of formula R R N-C = N- consists in the reaction of mitomycin C or mitomycin A or one of their N derivatives -substituted with an amide-acetal of formula:

  

  <EMI ID = 23.1>


  
where R, R and R are as defined above and R is a lower alkyl or cycloalkyl radical containing up to 6 carbon atoms or both

  
  <EMI ID = 24.1>

  
with the two oxygen atoms and the intermediate carbon atom a 5- or 6-membered ring structure. The reaction of these amide acetals with primary amines is conventional and it is the skill of the specialist to carry out the reaction with mitomycin C, mitomycin A or their Nla-alkyl derivatives. One can refer on this subject, for example, to H.E. Winberg patent of the United States of America n [deg.] 3,121,084 (February 11
1964) and to R.F. Abdulla et al. "The Chemistry of Formamide Acetals", Tetrahedron, volume 35 pages 1720-1724
(1979).

  
The Applicant prefers to carry out the reaction in a liquid anhydrous reaction medium in which the diluent is a liquid which is compatible with the reaction conditions. Preferably, this liquid is a lower halogenated aliphatic hydrocarbon or a lower alkanol or more advantageously a mixture

  
both. Chloroform and methanol as well as their mixtures are eminently suitable. The reaction is carried out at a temperature of 40 to 65 [deg.] C for a time sufficient for it to be completed.

  
When using a large excess (about 60 times) of acetal, the predominant product is

  
the bis-amidin product &#65533; that is to say a compound of formula I where A and B each represent the amidino radical. The N-formylated derivative sometimes appears as a by-product. However, when the quantity of acetal is limited (approximately 10 times), in addition to the bis-amine product, a mono-amine product is formed, namely a product of formula I where A represents the amino radical and B represents the radical. amidino. The mixtures of these reaction products are easily separated by chromatography as described in the examples below.

  
Various commercial amide-acetals which are suitable for this process are mentioned in Table I taken from the abovementioned article by Abdulla et al., Page 1685.

TABLE I

  
Various amide-acetals marketed

  

  <EMI ID = 25.1>


  
  <EMI ID = 26.1>

  
a cyano, di (lower alkyl) amino, lower alkoxy or lower alkylthio radical are obtained by replacing the amide-acetals in the above process with the following orthocarbonates:

  

  <EMI ID = 27.1>


  
These reagents can be obtained as described for that of formula VI by Kantlehner et al., Liebigs Ann. Chem., 1981, 70-84 and for those of formulas VII, VIII and IX by H. Meerwein, et al., Liebigs Ann. Chem.
641, 1 (1961).

  
The amine derivatives of formula I where represent

  
  <EMI ID = 28.1>

  
sense R NHC = N-, or R N = C-N- are obtained from

  
the anionic form of mitomycin C or one of its N7-substituted derivatives as described above. Halogenomethyliminium salts suitable for the process of the invention have been described in the literature. Examples are given by W. Kantlehner in "Advances in Organic Chemistry", volume 9, part 2, Wiley Interscience, 1079, pages 81 and 82. The table

  
II below is taken from Kantlehner's article.

  
Table II

Halogenomethyliminium salts

  

  <EMI ID = 29.1>
 

  

  <EMI ID = 30.1>
 

  
When a halogenomethyliminium salt

  
as illustrated in table II serves as a reagent, it is sometimes advantageous to use the corresponding amide as solvent, that is to say the amide from which

  
iminium salt. When the corresponding amides are solid, hexamethylphosphoramide or pyridine are also suitable. This is illustrated in Examples 17 and 19.

  
Imidoyl chlorides from N-substituted formamides are also suitable as reagents for this purpose. Their preparation is known, as it appears

  
  <EMI ID = 31.1>

  
of Imidoyl Halides ", Plenum Press, New York, 1968, pages 74-76. The reaction of these compounds with the amine-forming amines is also well known, as described by SR Sandler and W. Karo in" Organic Chemistry ", volume 12-111, AT Blomquist

  
and H. Wasserman, editors, Academic Press New York, page 227.

  
Table III

  
Imidoyl chloride
  <EMI ID = 32.1>
 Table III (continued)
  <EMI ID = 33.1>
 Table III (continued)

  

  <EMI ID = 34.1>


  
The compounds of formula I where A represents the

R <2>

  
amidino radical unsubstituted on nitrogen, H2N-C = N-,

  
R <2> R5

  
or HN = C-N- are obtained by reaction of an imino ether

  
  <EMI ID = 35.1>

  
in either anionic form as described above. The protective radical is then removed in a conventional manner. Isopropyl formimidate whose amino radical is protected by the radical

  
  <EMI ID = 36.1>

  
formula

  

  <EMI ID = 37.1>


  
the preparation methods consist in reacting the anionic form of mitomycin C with cyclic halomethyleneethylene salts or cyclic imidoyl halides where R2 and R <3> of the formula
  <EMI ID = 38.1>
 are united in one cycle. Suitable reagents

  
for the reaction with anionic mitomycin C are 2-chloro-1-methylpyridinium iodide (example 16),

  
2-chloro-4,5-dihydro-1-methyl-1 (3H) -pyrrolidinium chloride (Table II), N, N'-dimethylN, N'-trimethylenechloroformamidinium chloride (Example 28) and others cyclic imidoyl halides from 2-azetidinones, 2-pyrrolidinones, 2-piperidinones and 2-azepinones. Again, when R 7 or R9 in

  
the final product represents a hydrogen atom, the protective radical as above is used in

  
the cyclic halogenomethyleneiminium intermediate salt.

  
Compounds of formula I where A or A and B

  
represent an amidino radical of formula

  

  <EMI ID = 39.1>


  
react with primary amines of formula R5NH2

  
  <EMI ID = 40.1>

  
coyl, aryl or lower aralkyl each of which has up to 12 carbon atoms and heteroalicyclic or heteroaromatic radicals having 3 to 8 members, at least two of which are carbon atoms.

  
The only limitation weighing on the choice of the primary amine, other than the absence of functional radicals incompatible with the reaction conditions, is that the nitrogen atom of the amine function is united to a carbon atom which brings to the minus one hydrogen atom and less than two aryl radicals. An anhydrous liquid organic compound serves as a reaction medium and any such compound is suitable as long as it is compatible with the reaction conditions and does not adversely participate in the reaction. A molecular excess of the primary amine is generally used.

  
A reaction temperature of about -15 [deg.] C to +50 [deg.] C is preferred. The reaction product is a 7- (amino

  
  <EMI ID = 41.1>

  
tomycin C carrying a substituent as defined in

  
  <EMI ID = 42.1>

  
known to have significant activity against experimental tumors in animals.

  
Certain primary amines of formula R6NH2

  
have been found to be unsuitable for displacing the 7-amidino radical by the method described in the above paragraph. R6 represents an alkyl, cycloalkyl, cycloalkylalkyl, aralkyl or heteroalicyclic radical from 4 to

  
18 carbon atoms including the carbon atom bearing

  
the amino radical is a tertiary carbon atom

  
or a secondary carbon atom carrying two aryl radicals. Certain other weakly basic aliphatic amines, such as trifluoroethylamine also do not make it possible to displace the 7-amidino radical. These amines are useful for transforming a compound of formula I where A and B each represent the amino radical

  
formula dino

  

  <EMI ID = 43.1>


  
into a compound of formula I

  
where only A represents this amidino radical. These amines, although they do not have the possibility of displacing the 7-amidino radical, nevertheless have the capacity to split the amidino radical B into an NH2 radical to give rise to the carbamido function characteristic of unsubstituted mitosanes. The amine itself is suitable as a reaction medium or a solvent system of the type defined in the previous paragraph can be used. This process is preferably carried out at a temperature of 20 to 60 [deg.] C.

  
Melting points are determined using a ThomasHoover capillary melting point device and are not corrected. Temperatures are expressed in degrees Celcius. The proton nuclear magnetic resonance (NMR) spectra are recorded using a Varian XL100 spectrometer, in pyridine d5 unless otherwise indicated. Infrared spectra
(IR) are measured using a Beckman 4240 spectrophotometer on the sample mixed with bromide

  
  <EMI ID = 44.1>

  
using a Varian-Cary 219 spectrophotometer.

  
Thin layer chromatography (TLC) is carried out on plates pre-coated with 0.25 mm of silica gel which are observed in UV light. Rapid chromatography is performed on silica

  
  <EMI ID = 45.1>

  
reduced pressure below 50 [deg.] C.

  
EXAMPLE 1 -

  
Compound V

  
  <EMI ID = 46.1>

  
methylene-9a-methoxymitosan

  
Compound VI

  
  <EMI ID = 47.1>
  <EMI ID = 48.1>
 
  <EMI ID = 49.1>
 A total of 9.6 ml (2.4 ml fractions at 0, 18, 21 and 23 hours) of N, N-dimethylformamide dimethyl acetal is added to a suspension of 500 mg (1.50 mmol) of mitomycin C in 25 ml of chloroform and the suspension is stirred at about 50 [deg.] for 41 hours. After evaporation of the solvent and the excess reactants under reduced pressure, a dark green residue is obtained; TLC (methylene chloride / methanol 20: 1) reveals the absence of mitomycin C and the presence of two new green constituents (Rf = 0.16 and 0.22).

   The major constituent is isolated (Rf = 0.16) by rapid chromatography with elution using methylene chloride / methanol 20: 1 to obtain a green solid
(340 mg, 51.5%) giving, by dissolution in diethyl ether, then addition of hexane, compound V which is a dark green amorphous powder.

  
  <EMI ID = 50.1>

  
(s, 3H), 2.82 (s, 3H), 2.86 (s, 6H), 3.22 (s, 3H), 3.30
(if, 1H), 3.60 (d, J = 12Hz), 4.12 (dd, 1H, J = 10, 4Hz), 4.43 (d, 1H, J = 12Hz), 4.90 (if , 1H), 5.10 (t, 1H, J = 10 Hz), 5.52 (dd, 1H, J = 10, 4Hz), 7.85 (s, 1H), 8.64 (s, 1H) .

  
  <EMI ID = 51.1>

  
calculated C, 56.71; H, 6.08; N, 18.90% found C, 56.20; H, 6.28; N, 17.88% The minor constituent is isolated (Rf = 0.22)
(180 mg, 25.35%) as an amorphous solid by precipitation from diethyl ether and hexane and identified as Compound VI.

  
  <EMI ID = 52.1>

  
gulets, 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).

  
  <EMI ID = 53.1>

  
calculated C, 55.89; H, 5.93; N, 17.78%

  
found C, 55.41; H, 5.96; N, 16.99% TLC examination of the solutions of compounds V and VI in ethyl acetate or in dimethyl acetal

  
  <EMI ID = 54.1>

  
ambient for more than 10 hours reveals that compound VI (Rf = 0.22) is converted into compound V (Rf = 0.16) to give a solution highly enriched in the latter.

  
Examples 2 to 7 are carried out according to the procedure of Example 1 with the differences indicated below in order to obtain different other compounds in accordance with the invention.

  
EXAMPLE 2 -

  
Compound VII

  
  <EMI ID = 55.1>

  
amino) -methylene-9a-methoxymitosan

  
It is heated with stirring for 15 hours

  
at 53 [deg.] a suspension of mitomycin C (200 mg, 0.6 millimole) in diethylacetal of N, N-diisopropylformamide (3 ml). The reaction mixture is poured into 50 ml of water and extracted with ethyl acetate (3 x 30 ml).

  
  <EMI ID = 56.1>

  
they are evaporated to obtain a dark green syrup which

  
TLC (methylene chloride / methanol 10: 1) reveals

  
the presence of a major constituent at Rf = 0.43 in the presence of more mobile impurities (Rf = 0.45-0.50). The major constituent VII is isolated in the form of a dark green solid (156 mg, 46.8%) by two rapid chromatographies with elution using methylene chloride / methanol 20: 1.

  
  <EMI ID = 57.1>

  
2.78 (dd, 1H, J = 4.2Hz), 3.05 (d, 1H, J = 4Hz), 3.22 (s,

  
  <EMI ID = 58.1>

  
1H, J = 12 Hz), 4.56 (t, 1H, J = 10 Hz), 4.88 (dd, 1H, J = 10, 4Hz), 7.83 (s, 1H), 8.67 ( s, 1H).

  
  <EMI ID = 59.1>

  
1600, 1550, 1235, 1060.

  
UV (MeOH) At max, nm: 246 and 393

  
Analysis for C29H44N605.

  
calculated C, 62.55; H, 7.91; N, 15.10% found C, 62.03; H, 7.80; N, 14.60% EXAMPLE 3 -

  
Compound XIV

  
  <EMI ID = 60.1>

  
methylene-9a-methoxy-N -methylmitosan

  
In the present example, one starts from porfiromycin (Nla-methyl mitomycin C) in an amount of 130 mg,
(0.37 millimole) for the reaction with 0.8 ml (1.5 millimole) of N, N-dimethylformamide dimethylacetal in
10 ml chloroform and 2 ml methanol as solvent

  
  <EMI ID = 61.1>

  
isolates compound XIV in the form of a syrup after evaporation of the solvents and is purified by rapid chromatography on 20 g of silica gel which is eluted with methylene chloride / methanol (20: 1).

  
  <EMI ID = 62.1> (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, llHz), 4.38 (d, 1H, J = 12 Hz), 4.92 (t, 1H, J = llHz), 4.96 (sl, 1H), 5.46 (dd , 1H, J = 4, 11Hz), 7.86

  
(s, 1H), 8.70 (s, 1H).

  
TLC with methylene chloride / methanol 9: 1, Rf = 0.53.

  
  <EMI ID = 63.1>

  
calculated C, 57.60; H, 6.55; N, 18.33%

  
found C, 57.11; H, 6.11; N, 17.99%

  
In this way, compound XV or 7-amino-N-dimethylaminomethylene-9a-methoxy-N -methylmitosan is obtained in the form of a by-product.
30% TLC, Rf = 0.40 (methylene chloride / methanol 9: 1).

  
  <EMI ID = 64.1>

  
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, 10Hz), 4.50 (d, 1H, J = 10Hz), 4.90
(t, 1H, J = 10 Hz), 5.05 (sl), 5.43 (dd, 1H, J = 4, 10Hz), 8.70 (s, 1H).

  
  <EMI ID = 65.1>

  
1625, 1553, 1230, 1125.

  
UV (MeOH) &#65533; max, nm: 358, 244 and 216.

  
  <EMI ID = 66.1>

  
calculated C; 56.53; H, 6.20; N, 17.38% found C, 54.68; H, 6.13; N, 16.59% EXAMPLE 4 -

  
Compound IX

  
  <EMI ID = 67.1>

  
(1-piperidinylmethylene) mitosan

  
3 ml of N- (diethoxymethyl pip piperidine and 200 mg of mitomycin C at 60 [deg.]) Are reacted for 2.5 hours in solution in chloroform (3 ml).

  
The product is obtained with a yield of 27.6% whose TLC indicates an Rf of 0.20 (methylene chloride / methanol 20: 1).

  
  <EMI ID = 68.1>

  
2.80 (sl, 1H), 3.24 (s, 3H), 3.00-3.40 (m, 5H), 3.403.80 (m, 5H), 4.13 (dd, 1H, J = 10, 4Hz), 4.45 (d, 1H, J = 12 Hz), 4.90 (if, 2H), 5.12 (t, 1H, J = 10Hz), 5.56
(dd, 1H, J = 10 4Hz), 7.87 (s, 1H), 8.70 (s, 1H).

  
  <EMI ID = 69.1>

  
calculated C, 61.79; H, 6.87; N, 16.02%

  
found C, 61.01; H, 6.85; N, 15.34%

  
As a major constituent, the N-formylated derivative of the above compound is obtained, which is compound VIII or Nla-formyl-9a-methoxy-7 - [(1-piperidinylamino) -

  
  <EMI ID = 70.1>

  
with a yield of 43% and whose TLC indicates an Rf of 0.25 (methylene chloride / methanol 20: 1).

  
  <EMI ID = 71.1>

  
4.90 (t, 1H, J = llHz), 4.94 (si, 1H), 5.54 (dd, 1H, J = ll, 4Hz), 7.94 (s, 1H), 8.71 ( s, 1H), 9.08 (s, 1H).

  
  <EMI ID = 72.1>

  
Analysis for C28H36N606

  
calculated C, 60.08; H, 6.52; N, 15.21% found C, 59.99; H, 6.17; N, 15.07% EXAMPLE 5 -

  
Compound X

  
  <EMI ID = 73.1>

  
pholinomethylene) mitosan

  
A stirred suspension of mitomycin C (200 mg, 0.6 millimole) in chloroform (10 ml) and N-diethoxymethylmorpholine (4 ml) is heated to about 53 [deg.] For 42 hours. The reaction mixture is concentrated to a syrup under high vacuum. Separation is carried out by summary rapid chromatography
(methylene chloride / methanol 25: 1) to isolate green compounds from excess reagents. All the green compounds are dissolved in 20 ml of ethyl acetate and the solution is washed with water (3 x 20 ml). The mixture is extracted from the washing water again with ethyl acetate (3 x 15 ml). After combining all the fractions in ethyl acetate, they are dried (Na2SO4) and evaporated to obtain a dark green syrup whose TLC (methylene chloride / methanol 10: 1) reveals a defined green constituent at

  
Rf = 0.33, in addition to various green impurities (Rf = 0.35 to 0.40). By rapid chromatography, the compound of Rf = 0.33 (130 mg, 56.8%) which is a dark green amorphous solid identified as compound X is isolated.

  
  <EMI ID = 74.1>

  
1H, J = 2Hz), 3.22 (s, 3H), 3.30-3.94 (m, 18H), 4.20 (d, 1H, J = 12Hz), 4.40 - &#65533; sl, 1H), 4.54 (t, 1H, J = 10Hz), 4.88
(dd, 1H, J = 10Hz, 4Hz), 7.74 (s, 1H), 8.51 (s, 1H).

  
  <EMI ID = 75.1>

  
calculated C, 56.78; H, 6.06; N, 15.90%

  
found C, 53.07; H, 6.03; N, 15.37% EXAMPLE 6 -

  
Compound XVI

  
7-Amino-N-dimethylaminomethylene-9a-methoxymitosan

  
Mitomycin C (200 mg, 0.6 millimole) is dissolved in 10 ml of chloroform and 2 ml of methanol and 0.64 ml of N, N-dimethylformamide dimethylacetal (4.8 mmol) are added, then the solution is stirred at about 50 [deg.] for 50 minutes. TLC (methylene chloride: methanol 90:10) reveals a trace of unreacted mitomycin C (Rf = 0.22) and two new constituents (Rf = 0.42 and 0.33 respectively). The solution is concentrated under reduced pressure in a syrup which is subjected to rapid chromatography (25 g

  
silica gel) with elution with methylene chloride / methanol (20: 1).

  
The most mobile constituent is isolated (Rf = 0.42) in the form of a green solid (60 mg, 22.5%)

  
  <EMI ID = 76.1>

  
being compound V.

  
The major blue constituent is isolated (Rf = 0.33) in the form of an amorphous solid (148 mg, 63.3%) which is characterized as being compound XVI. An analytical sample is prepared by precipitation from methylene chloride and n-pentane.

  
  <EMI ID = 77.1>

  
(s, 3H), 3.21 (s, 3H), 3.28 (d, 1H, J = 4Hz), 3.62 (dd, 1H, J = 2, 13Hz), 3.94 (si), 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, 10Hz).

  
  <EMI ID = 78.1>

  
calculated C, 55.48; H, 5.91; N, 17.98% found C, 54.70; H, 6.14; N, 17.95% EXAMPLE 7 -

  
Compound XVII

  
  <EMI ID = 79.1>

  
The mitomycin C of Example 1 is replaced by mitomycin A (170 mg) which is reacted with the dimethylacetal of N, N-dimethylformamide (0.6 ml) in solution in chloroform / methanol (10: 1) at 50 [deg.] for 1 hour. The desired compound is obtained with a yield of 48%, the TLC of which indicates an Rf of 0.50
(methylene chloride / methanol 9: 1).

  
  <EMI ID = 80.1>

  
(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

  
  <EMI ID = 81.1>

  
calculated C, 56.39; H, 5.94; N, 13.85%

  
found C, 56.51; H, 5.92; N, 13.71%

  
The Nla-formylated derivative of compound XVII, that is to say the compound, is obtained with a yield of 16.5%.

  
  <EMI ID = 82.1>

  
b formylmitosan, whose TLC indicates an Rf of 0.61 (methylene chloride / methanol 9: 1).

  
  <EMI ID = 83.1>

  
(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 XIX

  
7- (Dimethylaminomethylene) amino-9a-methoxymitosan

  
Aminodiphenylmethane (2.2 ml,
10.8 millimoles) to 600 mg of compound V (1.35 millimoles) dissolved in methanol (10 ml) and the solution is stirred at 54 [deg.] For 4 hours. The progress of the reaction is observed by TLC (methylene chloride / methanol 90:10). At the end of the 4 hours, the starting compound (Rf = 0.35) has disappeared and a new important green zone (Rf = 0.29) has appeared. The solution is concentrated under reduced pressure and the resulting syrup is subjected to rapid chromatography (25 g of silica gel) with elution using methylene chloride / methanol 20: 1. We gather the fractions containing

  
the green compound (Rf = 0.29), they are dried (Na2SO4) and

  
we concentrate them. The compound XIX is isolated in the form of an amorphous solid (215 mg, 41%).

  
  <EMI ID = 84.1>

  
(s, 3H), 2.88 (s, 3H), 3.08 (si, 1H), 3.24 (s, 3H), 3.56
(if, 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
(if, 2H), 7.88 (s, 1H).

  
  <EMI ID = 85.1>

  
calculated C, 55.48; H, 5.91; N, 17.98%

  
found C, 54.83; H, 5.67; N, 16.90%

  
By replacing compound V in Example 8 as the starting compound with the N-formylated derivative or compound VI and carrying out the reaction for 20 hours at room temperature, compound XIX is obtained in substantially the same way and with the same yield.

  
EXAMPLE 9 -

  
Compound XX

  
7- (Dimethylaminomethylene) amino-9a-methoxy-Nla-methylmitosan

  
1 g of compound XIV (2.18 mmol) is dissolved in methanol (20 ml) and aminodiphenylmethane (3.5 ml, 17.18 mmol) is added, then the resulting solution is stirred for 5 hours at room temperature and for 5 hours at 40 [deg.]. The TLC (CH2C12MeOH 90:10) of the reaction mixture reveals that the major part of the starting compound (Rf = 0.55) has been consumed and that a significant new green zone has formed (Rf = 0.48 ). Continuation of operations as in Example 8 leads to compound XX

  
which is an amorphous solid (350 mg). The purification is completed by rapid chromatography (7 g of silica gel) with elution using methylene chloride / methanol (250 ml, 96/4 v / v) and by precipitation of the resulting solid (Rf = 0.48) at starting from methylene chloride (5 ml) and hexane (50 ml) to isolate the analytically pure compound XX (314 mg, 35.7%) which is a solid.

  
  <EMI ID = 86.1>

  
10Hz), 4.17 (d, 1H, J = 12Hz), 4.38 (t, 1H, J = 10Hz), 4.68 (m, 2H), 4.76 (dd, 1H, J = 4, 10Hz), 7.72 (s, 1H).

  
  <EMI ID = 87.1>

  
calculated C, 56.53; H, 6.20; N, 17.36%

  
  <EMI ID = 88.1>

  
EXAMPLE 10 -

  
Compound XI

  
7- (n-Propyl) amino-9a-methoxymitosan

  
330 mg of compound V (0.74 millimole) are dissolved in anhydrous methanol (10 ml) and n-propylamine (1.0 ml) is added thereto. The reaction mixture is stirred for 6 hours at room temperature

  
  <EMI ID = 89.1>

  
solvent and excess reagents under reduced pressure and the residue is subjected to rapid chromatography

  
on silica gel. We reprecipitate the blue constituent
(Rf = 0.40), obtained by elution with methylene chloride / methanol 30: 1, from methylene chloride with hexane to obtain compound XI which is an amorphous gray powder (125 mg, 44 , 5%).

  
  <EMI ID = 90.1>

  
(s, 3H), 2.74 (sl, 1H), 3.12 (sl, 1H), 3.22 (s, 3H), 3.36 (q, 2H), 3.60 (d, 1H, J = 12Hz), 3.96 (dd, 1H, J = llHz, 4Hz), 4.54 (d, 1H, J = 12Hz), 5.00 (m, 3H), 5.36 (dd, 1H, J = 11.4 Hz), 6.90 (t, 1H).

  
  <EMI ID = 91.1>

  
calculated C, 57.40; H, 6.38; N, 14.88%

  
found C, 57.28; H, 6.41; N, 14.08%

  
EXAMPLE 11 -

  
Compound XII

  
7- (2-Hydroxyethyl) amino-9a-methoxymitosan

  
330 mg of compound V (0.74 millimole) are dissolved in anhydrous methanol (5 ml) and ethanolamine (2 ml) is added. The reaction mixture is stirred at room temperature for 2 hours, then diluted with water (50 ml) and extracted with ethyl acetate (5 x 60 ml). The mixture of extracts in ethyl acetate (Na2SO4) is dried and concentrated to a bluish purple residue, giving, by column chromatography with elution using 10% methanol in methylene chloride and by concentration of the fractions pool containing compound XII, 105 mg
(37%) of compound XII. Which is an amorphous solid.

  
  <EMI ID = 92.1>

  
(d, 1H, H = 4Hz), 3.24 (s, 3H), 3.65 (dd, 1H, J = 2, 12Hz), 3.70-4.20 (m, 5H), 4.52 (d, 1H, J = 13Hz), 4.96 (t, 1H, J = 12Hz), 7.38 (t, 1H), 7.58 (if).

  
  <EMI ID = 93.1>

  
Analysis for C17H22N406

  
calculated C, 53.92; H, 5.82; N, 14.80%

  
found C, 51.30; H, 5.88; N, 14.80%

  
EXAMPLE 12 -

  
Compound XIII

  
7- [2-Benzylthioethyl] amino-9a-methoxymitosan

  
200 mg of compound V (0.45 millimole) are dissolved in methanol (2 ml) and S-benzyl2-aminoethanethiol (0.5 ml) is added, then the solution is stirred at room temperature for 16 hours. The residue which the evaporation of the solvent gives under reduced pressure is subjected to rapid chromatography (40 g of silica gel) with elution using 6% methanol in methylene chloride (400 ml). The blue constituent is isolated (Rf about 0.5 in methanol at
10% in methylene chloride) as an amorphous solid (65 mg, 29.8%). The spectroscopic data (NMR, IR, UV and mass) are compatible with the assigned structure.

  
  <EMI ID = 94.1>

  
calculated C, 59.49; H, 5.82; N, 11.56% found C, 59.72; H, 5.94; N, 11.08%
  <EMI ID = 95.1>
(A) Diisopropylethylamine is added slowly
(2.1 millimoles) at 0 [deg.] In a nitrogen atmosphere to a solution of isopropyl formimidate hydrochloride
(1 millimole) in dimethylformamide (DMF). To the resulting solution is added dropwise to 0 [deg.] <EMI ID = 96.1> resulting limpid tion is the solution'A.

  
(B) Add a solution of mitomycin C (1 millimole) in 5 ml of DMF to a suspension of sodium hydride
(1.5 millimole) in 3 ml of DMF. The suspension was stirred at room temperature for 20 minutes and cooled to about -40 to -50 [deg.] Before adding solution A above. We keep the solution at
-40 [deg.] For 1 hour, then allowed to warm to room temperature. After standing at room temperature for about 6-18 hours, the reaction mixture is diluted with methylene chloride and filtered.

   The solid residue which the evaporation of the filtrate gives is subjected to chromatography on silica gel to isolate the desired compound whose amidino function is protected.
(C) The protective radical of the amidino function of the above intermediate is eliminated according to the method described by Carpino and Tsao (J. Chem. Soc. Chem. Comm.
358 (1978)) so as to obtain the desired unsubstituted amidine.
  <EMI ID = 97.1>
(A) Diisopropylethylamine (21 millimoles) is added slowly at 0 [deg.] Under a nitrogen atmosphere to a solution of isopropyl formimidate hydrochloride (1 millimole) in DMF (2 ml). Methyl iodide is added to <EMI ID = 98.1>

  
solution B.

  
(B) The operations of Example 13 (B) are repeated, replacing solution A with solution B in order to obtain the desired compound.

  
EXAMPLE 15 -

  

  <EMI ID = 99.1>


  
compound XIX

  
A 0.5 M solution of N, N-dimethylchloromethyleniminium chloride is prepared by adding drop

  
oxalyl chloride (1.57 g, 12.5 millimoles)

  
  <EMI ID = 100.1>

  
in 25 ml of chloroform, then stirring the mixture for 30 minutes at room temperature. Separately, a solution of mitomycin C (334 mg,

  
1 millimole) in 5 ml DMF to a NaH suspension
(36 mg, 1.5 millimole) in 3 ml of DMF. The solution is stirred at room temperature for 20 minutes

  
and it is cooled to about -40 to -50 [deg.], then the above solution of N, N-dimethylchloromethyleniminium chloride (3 ml, 1.5 millimole) is added to it. An additional NaH (18 mg, 0.75 millimole) is added after 10 minutes of stirring at -40 [deg.]. The solution is stored at -40 [deg.] For 1 hour, then it is diluted with methylene chloride and filtered. The residue which the evaporation of the filtrate gives is subjected to TLC on silica gel (elution with 10% methanol in methylene chloride). The extraction of the main green strip gives
78 mg (43% based on the mitomycin C collected) of an amorphous solid whose NMR spectrum and TLC are identical to those of compound XIX obtained in Example 8. The extraction of the purple band gives 150 mg of mitomycin C.

  
EXAMPLE 16 -

  
7- (1-Methyl-2- (H) -pyridinylidene) amino-9a-methoxymitosan

  

  <EMI ID = 101.1>


  
  <EMI ID = 102.1>

  
C (242 mg, 0.725 millimole) and NaH (43.5 mg, 1.81 millimole). After 15 minutes of stirring, 2-chloro-1-methylpyridinium iodide (370 mg, 1.45 millimole) is added at room temperature. The solution is stirred for 1.5 hours and then diluted with ethyl acetate, then filtered. The residue which the evaporation of the filtrate gives is subjected to TLC on silica gel (elution with 5% methanol in chloride

  
methylene). The minor product (12 mg) is compound XIX (Example 8). The major product (75 mg) is purified by TLC on silica gel (10% methanol in methylene chloride) to collect 6 mg (2%) of the desired compound.

  
  <EMI ID = 103.1>

  
5.10 (t, 1H, J = 10 Hz), 5.43 (dd, 1H, J = 10, 4Hz), 5.99 (dt, 1H, J = 9.2 Hz), 6.09 (dd , 1H, J = 9.1 Hz), 6.95 (dd, 1H, J = 9, 7.2 Hz), 7.32 (dd, 1H, J = 7.1 Hz). EXAMPLE 17 -

  
&#65533; [(Methylaminomethylene) amino] -9a-methoxymitosan

  

  <EMI ID = 104.1>


  
Sodium hydride (12 mg, 0.5 millimole) is added under a nitrogen atmosphere to a solution of mitomycin C (167 mg, 0.5 millimole) in 2 ml of hexamethylphosphoramide. To this solution is added N-methylformimidoyl chloride (19 mg, 0.25 millimole), N.H. Bosshard and H. Zollinger, Helv. Chim. Acta, 42,
1659 (1959). The solution is stirred at room temperature for 10 minutes, then sodium hydride (6 mg, 0.25 millimole) and N-methylformimidoyl chloride (9.5 mg, 0.13 millimole) are added thereto. After 6 to 12 hours of stirring, the solution is diluted with ethyl acetate and filtered. By evaporation of the solvent and chromatographic purification of the residue, the desired compound is obtained.

  
EXAMPLE 18 -

  
Compound XXI

  
9a-Methoxy-7- (1-morpholinomethylene) aminomitosan

  

  <EMI ID = 105.1>


  
4-diethoxymethylmorpholine is added
(12.5 ml) to mitomycin C (600 mg, 1.8 millimole) suspended in chloroform (30 ml) and the resulting suspension is heated to 58 [deg.] For 48 hours. After 48 hours, the TLC (20% methanol in methylene chloride) reveals that the reaction is not completed. The solution is concentrated under reduced pressure to obtain a syrup to which water is added
(100 ml). After 20 minutes of stirring, the dark green solution is extracted with methylene chloride (5 × 50 ml.) And the combined extracts are dried and concentrated to

  
a syrup. Aminodiphenylmethane (6.5 ml) is added to the syrup in methanol (20 ml) and the solution is stirred-EXAMPLE 23 -

  
Compound XXV

  
  <EMI ID = 106.1>

  

  <EMI ID = 107.1>


  
Heated to 55 [deg.] For 5 hours 1.5 g of 2,2-dimethoxy-1-methylpyrrolidine (10.3 millimoles)

  
(H. Eilingsfeld et al. Angew. Chem., 72, 836 (1960)), and 280 mg of mitomycin C (0.34 millimole) in 20 ml of methanol. The reaction mixture was examined by thin layer chromatography on an alumina plate with methylene chloride / methanol 97: 3 as solvent. The TCC reveals a main green spot corresponding to the product and a minor blue spot corresponding to the starting mitomycin C. The solvent is distilled off under vacuum at 40 [deg.] And the residue is dissolved in methylene chloride, then the solution is poured onto a 4.5 cm column containing 150 g of alumina. The elution is carried out with 50 ml of methylene chloride, then 600 ml of 1% methanol in

  
methylene chloride. Large impurities are removed, but without isolating pure fractions. The combined eluate is concentrated by distillation at 20 [deg.] To an oily residue which appears to contain a certain amount of 2,2-dimethoxy-1-methylpyrrolidine. The substance is again chromatographed on an alumina column (25 g) using 200 ml of methylene chloride, then 100 ml of 1% methanol in methylene chloride. The 2,2-dimethoxy-1methylpyrrolidine is thus eliminated in order to obtain a certain number of fractions containing minor impurities and some pure fractions confirmed by TLC (a single green spot) providing the desired product, yield 53 mg.

  
  <EMI ID = 108.1>

  
calculated C, 58.66; H, 6.64; N, 16.42% found C, 58.63; H, 6.46; N, 16.50% UV (MeOH) max max. nm: 354, 239

  
  <EMI ID = 109.1>

  
compatible with the structure of the compound.

  
EXAMPLE 24 -

  
7 - [(1-Methyl-2-pyrrolidinylidene) aminoJ-9a-methoxymitosan

  

  <EMI ID = 110.1>


  
A solution of 80 mg (0.16 millimole) of compound XXV and 0.48 ml of n-butylamine in 15 ml of chloroform is heated at reflux for 48 hours.

  
The TLC (methanol / methylene chloride, 2% on alumina) reveals a main green spot, a small more advanced blue spot and a small more late red spot which are due to the starting compound. The reaction solution is poured onto a column containing 50 g of alumina which is eluted with 200 ml of 1% methanol in methylene chloride, then 400 ml of 2% methanol in methylene chloride. The fractions containing a single main green constituent revealed by the TLC are combined and concentrated in a residue which is the desired product with a weight of 24 mg.

  
  <EMI ID = 111.1>

  
(q, 2H), 2.72 (sl, 1H), 2.84 (s, 3H), 3.12 (m, 3H), 3.24 (s, .3H), 3.60 (dd, 1H , J = 14.2 Hz), 4.00 (dd, 1H, J = 12.6 Hz), 4.40 (d, 1H, J = 14 Hz), 5.04 (t, 1H, J = 14 Hz), 5.38 (dd, 1H, J = 12.6 Hz), 7.48 (sl, 2H). EXAMPLE 25 -

  
Compound XXVI

  
7 - [(Methoxyamino) methylene] amino-9a-methoxymitosan

  

  <EMI ID = 112.1>


  
A solution of 660 mg of compound XIX (1.7 millimole) in 10 ml of methanol is prepared and 170 mg of methoxyamine hydrochloride (2.0 millimoles) are added thereto. The solution is stirred at 10 [deg.] For 3 hours and at room temperature for 2 hours. The TCC reveals only a trace of the unreacted compound XIX. A black precipitate is formed on standing, which is collected and washed with acetone to obtain 380 mg of the desired product (57%).

  
  <EMI ID = 113.1>

  
calculated C, 52.19; H, 5.40; N, 17.90% found C, 51.64; H, 5.40; N, 17.83%

  
  <EMI ID = 114.1> compatible with the structure of the announced compound or that of its C-7 tautomer, namely

  

  <EMI ID = 115.1>


  
EXAMPLE 26 -

  
Compound XXVII

  
7 - [(Benzyloxyamino) methylene] amino-9a-methoxymitosan

  

  <EMI ID = 116.1>


  
A solution of 100 mg of compound XIX (0.26 millimole) in 2 ml of methanol containing 0.5 ml of triethylamine is prepared and 400 mg (2.5 millimoles) of O-benzylhydroxylamine hydrochloride are added thereto. The reaction is allowed to progress for 2.5 hours at

  
Room temperature. The TLC (methylene chloride / methanol 10: 1) reveals a main orange brown zone in front of the green zone, which is that corresponding to compound XIX. The reaction mixture is concentrated to a residue which is subjected to rapid chromatography on silica gel (20 g) using methylene chloride / methanol 20: 1 for elution. The main brown zone is collected providing the desired product which is collected in the form of an amorphous solid with a weight of 80 mg (yield of 65.6%).

  
  <EMI ID = 117.1>

  
calculated C, 59.10; H, 5.35; N, 14.97% found C, 58.43; H, 5.48; N, 14.62%

  
  <EMI ID = 118.1> 1570, 1275, 1220, 1060.

  
  <EMI ID = 119.1>

  
compatible with the structure of the compound announced or that of its C-7 tautomer, namely

  

  <EMI ID = 120.1>


  
10 mg of compound XIX which has not reacted are recovered.

  
EXAMPLE 27 -

  
Compound XXVIII

  
7- (1,3-Dimethyl-2-imidazolinylidene) -9a-methoxymitosan

  

  <EMI ID = 121.1>


  
0.34 g of mitomycin C (1 millimole) is dissolved in 5 ml of 1,3-dimethyl-2-imidazolidone and 0.1 g of sodium hydride is added thereto at room temperature

  
(50% in oil, 2.08 millimoles). The mixture is stored for 20 minutes at room temperature, then cooled in an ice and salt bath (-15 [deg.]

  
The mixture is stored for 10 minutes at this temperature, then 0.65 g (2 millimoles) of 2-chloro-1,3-dimethyl-4,5-dihydro- (3H) -imidazoliminium chloride is added thereto. The mixture is kept at -15 [deg.] For

  
1 hour, then diluted with ethyl acetate and

  
chromatography on an alumina column. The column is eluted with methylene chloride, then with methylene chloride containing 2% by volume of methanol. A green fraction is collected consisting of the desired product which is further purified by chromatography on alumina which is eluted with methylene chloride containing 10% by volume of methanol, production 20 mg (5%).

  
  <EMI ID = 122.1>

  
calculated C, 53.03; H, 6.34; N, 18.55% found C, 52.68; H, 6.21; N, 18.15%

  
  <EMI ID = 123.1>

  
(s, 3H), 2.74 (m, 1H), 3.03-3.32 (m, 5H), 3.26 (s, 3H), 3.66 (dl, 1H, J = 12 Hz) , 4.02 (dd, 1H, J = ll, 4 Hz), 4.75 (d, 1H, J = 12 Hz), 5.09 (tl, 1H, J = ll Hz), 5.44

  
  <EMI ID = 124.1>

  
IR (KBr) 3400, 3280, 2930, 1700, 1610, 1480, 1330,

  
  <EMI ID = 125.1>

  
EXAMPLE 28 -

  
Compound XXIX

  
7 - (1,3-Dimethyltetrahydropyrimidinylidene) amino-9amethoxymitosan

  

  <EMI ID = 126.1>


  
Sodium hydride (50% dispersion in oil, 200 mg, 4.2 millimoles) is added in a nitrogen atmosphere to a solution of mitomycin C (680 mg, 2 millimoles) in 8 ml of 1, 3-dimethyl-3,4,5,6-tetrahydro (1H, 3H) -2-pyrimidinone. The mixture is stored for 20 minutes at room temperature, then

  
it is cooled to -25 [deg.]. 0.73 g of 2-chloro-1,3-dimethyl-2,3,4,5-tetrahydro-pyrimidinium chloride (4 millimoles) is added thereto and the new mixture is stored at -25 [deg.] For 3 hours. It is then diluted with ethyl acetate and 2 ml of methanol. The mixture is poured without further treatment onto a dry column of chromatographic alumina which is eluted first with methylene chloride, then with 2% methanol in methylene chloride to obtain 0.35 g of the desired compound (yield 39.5%),

  
P.F. 138-140 [deg.].

  
  <EMI ID = 127.1>

  
calculated C, 54.65; H, 6.33; N, 18.21% found C, 54.78; H, 6.18; N, 18.21%

  
  <EMI ID = 128.1>

  
(s, 3H), 2.64 (s, 3H), 2.76 (m, 1H), 2.90-3.30 (m, 5H), 3.26 (s, 3H), 3.74 ( d, 1H, J = 12Hz), 4.05 (dd, 1H, J = ll, 4 Hz), 4.97 (d, 1H, J = 12 Hz), 5.09 (t, 1H, J = ll Hz), 5.41 (dd, 1H, J = ll, 4 Hz).

  
IR (KBr) 3430, 3280, 2930, 1710, 1570, 1480, 1450,

  
  <EMI ID = 129.1>

  
EXAMPLE 29 -

  
Compound XXX

  
7- (Tetramethyldiaminomethylene) amino-9a-methoxymitosan

  

  <EMI ID = 130.1>

V

  
425 mg of mitomycin C (1.42 millimole) is mixed with 85.3 mg of sodium hydride in dispersion

  
at 50% in oil and 4 ml of dimethylformamide are added thereto. The mixture is stirred at room temperature in an argon atmosphere for 10 minutes, then cooled to -35 [deg.]. 289 mg of tetramethylchloroformamidinium chloride (2.13 millimoles) are added and

  
the mixture is allowed to warm up to 5 [deg.]

  
2 hours. Then add crushed dry ice

  
to the mixture to stop the reaction and the solvent is distilled off under reduced pressure. The residue is chromatographed on a column of 100 g of alumina which is eluted with 3% by volume methanol in methylene chloride. The compound is further purified by TLC on alumina (5% methanol by volume in methylene chloride) to obtain two fractions of 17 mg and 76 mg. The latter is recrystallized from acetone-ether to obtain the desired product, mp 193-195 [deg.], (Yield 12%).

  
  <EMI ID = 131.1>

  
calculated C, 55.54; H, 6.53; N, 19.43% found C, 54.92; H, 6.53; N, 19.29%

  
  <EMI ID = 132.1>

  
(s, 6H), 2.75 (m, 1H), 3.15 (d, 1H, J = 4 Hz), 3.26 (s, 3H), 3.65 (d, 1H, J = 12 Hz ), 4.00 (dd, 1H, J = ll, 5 Hz), 4.62 (d, 1H, J = 12 Hz), 5.04 (t, 1H, J = ll Hz), 4.38
(dd, 1H, J = 11.5 Hz).

  
IR (KBr) 3430, 3280, 2920, 1710, 1610, 1495, 1335,

  
  <EMI ID = 133.1>

  
EXAMPLE 30 -

  
Compound XXXI

  
  <EMI ID = 134.1>
  <EMI ID = 135.1>
 A 0.5 M solution of piperidinylchloromethyleniminium chloride is prepared by adding drop

  
oxalyl chloride (380 mg, 3 millimoles)

  
to 6 ml of chloroform containing 0.34 g (3 millimoles) of 1-formylpiperidine. Sodium hydride (96 mg) (50% oil dispersion, 2 millimoles) is added separately in a nitrogen atmosphere to a solution of mitomycin C (334 mg, 1 millimol.e) in 3 ml of 1-formylpiperidine. After 15 minutes of stirring at room temperature, the solution is cooled to -25 [deg.]

  
and there is added the iminium solution prepared previously (4 ml, 2 millimoles). The reaction mixture is kept at -25 [deg.] For 1 hour and the reaction is stopped by adding dry ice. After addition of methanol (1 ml), the product is absorbed on neutral alumina. The loaded alumina is deposited on a column of 30 g of alumina. The column is eluted first with methylene chloride, then with 3% by volume of methanol in methylene chloride to obtain 360 mg (84%) of the desired compound, P.F. 68-70 [deg.].

  
  <EMI ID = 136.1>

  
calculated C, 55.80; H, 6.58; N, 15.49% found C, 55.57; H, 6.21; N, 15.91%

  
  <EMI ID = 137.1>

  
(m, 1H), 3.06-3.30 (m, 3H), 3.25 (s, 3H), 3.48-3.70

  
(m, 2H), 3.57 (d, 1H, J = 13 Hz), 4.01 (dd, 1H, J = ll, 4 Hz). 4.43 (d, 1H, J = 13 Hz), 5.02 (tl, 1H, 3 = 11 Hz), 5.55 (dd, 1H, J = ll, 4 Hz), 7.86 (s, 1H).

  
IR (KBr) 3440, 3350, 3300, 2935, 2835, 1710, 1615, 1520,

  
  <EMI ID = 138.1>

  
212 (shoulder) nm.

  
EXAMPLE 31 -

  
7-Hydroxy-N-dimethylaminomethylene-9a-methoxymitosan
  <EMI ID = 139.1>
 Dimethylformamide dimethylacetal (1 ml) is added to a solution of 7-hydroxy-9a-methoxymitosan (20 mg) in methylene chloride (3 ml)

  
and the solution is stirred at about 65 [deg.] for 30 minutes. The progress of the reaction is observed by TLC
(methylene chloride / methanol 10: 1). The product is collected by concentrating the mixture under reduced pressure and by chromatographing the residue on silica gel to isolate the desired compound.

  
Table IV collates the test results.

  
  <EMI ID = 140.1>

  
those to which a tumor inoculum of 106 mouse P-388 leukemia ascites cells was intraperitoneally implanted and to which different doses of an experimental compound of formula I or of mitomycin C were administered. administered by intraperitoneal injection. For each dose, groups of six mice are taken and given a single dose of the compound once a day. For each series of experiments, a group of ten control mice receiving saline physiological solution is taken. The positive control is formed by the groups of mice receiving mitomycin C. A 30-day program is applied and the average survival in days is determined in each group of mice, as well as the number of mice surviving after 30 days .

   The mice are weighed before treatment and again on the sixth day. We assess the toxicity

  
of the active ingredient based on the evolution of weight loss. We choose mice weighing 20 g and

  
weight loss of up to 2 g is considered not to be excessive. We express the results under

  
the form% T / t which is the ratio of the average survival time in the treated group to the average survival time in the control group receiving the physiological saline solution, which multiplies 100. Usually, the animals receiving the physiological saline solution die in the nine days. The "maximum effect" in the table below is indicated in% T / t and the dose ensuring this effect is specified. Values in parentheses

  
are those obtained with mitomycin C during

  
of the same experience on the positive control. It is therefore possible to estimate the toxicity of the compounds of the invention by comparison with that of mitomycin C. It is considered that a minimum effect in terms

  
% T / t is 125. The minimum effective dose mentioned in the table below is the dose ensuring a% T / t of approximately 125. The two values specified in each case in the column entitled "average weight change" are respectively the average change of

  
weight per mouse at maximum effective dose and minimum effective dose.

TABLE IV

  
Inhibition of mouse P-388 leukemia

  

  <EMI ID = 141.1>
 

TABLE IV (continued)

  
Inhibition of mouse P-388 leukemia

  

  <EMI ID = 142.1>


  
 <1> in mg / kg bodyweight

  
 <2> in g per mouse, on days 1 and 6 at the maximum and minimum effective doses

  
Compounds XIX and XX are exceptionally interesting because their activity clearly outweighs that of mitomycin C both by the maximum effect and by the power in mg (importance of the doses ensuring equivalent effects). These are each of the compounds of formula I where A represents the radical

  
  <EMI ID = 143.1>

  
  <EMI ID = 144.1>

  
  <EMI ID = 145.1>

  
an aminomethylene radical of the formula R <3> R4N-C =

  
  <EMI ID = 146.1>

  
The bis-amidines of formula I forming the subject of the invention where A and B each represent such an amidino radical are also of significant interest by their activity against tumors. In this connection, reference can be made to the data specified in the table for the compounds V, VI, VII, IX, X and XIV which have this structural characteristic.

  
Table V collates the results of tests against melanoma B16 maintained in mice. We

  
  <EMI ID = 147.1>

  
peritoneally an implant of the tumor. The program lasts 60 days. Groups of ten mice are taken for each dose and the time is determined

  
average survival in each group. Control animals inoculated as test animals and receiving the injection vehicle without active ingredient

  
have an average survival time of 21 days. Efficacy is evaluated by comparing the survival time with that of the controls (% T / t) and the maximum effective dose and the minimum effective dose are determined for each compound. The minimum effective dose is that giving

  
a% T / t of 125. For each dose, the animals are administered the test compound on days 1, 5 and 9 intraperitoneally. The toxicity is evaluated by measuring the average change in weight on the day indicated, at the maximum effective dose and at the minimum effective dose. A 2 g weight loss for a 20 g mouse is not excessive.

TABLE V

  
Inhibition of melanoma B16 Change

  

  <EMI ID = 148.1>


  
* The values in brackets are those of the controls receiving mitomycin C.

  
Compound XXX (example 29) and compound XXIX (example 28) are tested on mouse B16 melanoma after subcutaneous implantation of the tumor

  
and by intravenous administration of the active ingredient. The survival time is determined as above during a 40-day treatment. We note the change in weight on the 12th day. The maximum effective dose of compound XXX is 1 mg per kg and provides a% T / t of 156 with a weight gain of 1.5 g. Groups of six animals are used, of which three animals survive for 40 days with this dose. The minimum effective dose is 0.25 mg / kg, for which the change in weight on day 12 is 1.0 g. The maximum effective dose of compound XXIX is 8 mg / kg with a% T / t

  
of 177 and a change in weight of -0.6. The minimum effective dose is 4 mg / kg with a weight change of +0.8. In the same experiment, the maximum effective dose of mitomycin C is 3 mg / kg with a% T / t of 195 and a change in weight of -0.5. The minimum effective dose of mitomycin C has not been determined.

  
During a rapid toxicological study on groups of five male BDFl mice per dose receiving a single intraperitoneal dose of compound XIX, no significant reduction in the number of lymphocytes was observed at the optimal effective dose of this compound
(1.6 mg / kg i.p.). At this dose, there is no appreciable increase in blood uric nitrogen or serum glutamic phosphotransferase, which reveals the absence of effect on renal or hepatic function or the suppression of activity. lymphocyte.

  
Because of the remarkable activity manifested against experimental tumors in animals and of a lower toxicity than that of mitomycin C, the invention also relates to the administration of the compounds of formula I for inhibiting tumors in mammals. To this end, the compounds can be administered systemically in an amount which is substantially non-toxic, but effective against tumors, to an animal carrying a tumor.


    

Claims (1)

R E V E N D I C A T I O N S 1 - Compound of formula:  <EMI ID = 149.1> where A represents an amino, methoxy, hydroxyl radical, (1-lower alkyl-2 (1H) -pyridinylidene) amino or a radical of formula:  <EMI ID = 150.1> B represents an amino radical or the amidino radical of formula  <EMI ID = 151.1> and at least one of A and B represents one of the specified radicals other than amino, methoxy or hydroxyl, n represents 0, 1, 2 or 3, R1 represents a hydrogen atom or lower alkyl, lower alkanoyl, benzoyl or substituted benzoyl radical, the substituent of which is a lower alkyl, lower alkoxy, halo, amino or nitro radical, <EMI ID = 152.1> lower alkyl, phenyl, (lower alkyl) phenyl, (lower alkoxy) phenyl, halophenyl, aminophenyl, nitrophenyl, thienyl, furyl, cyano, di (lower alkyl) amino, lower alkoxy or lower alkyl, R <3> represents a lower alkyl radical, lower alkoxy or constitutes with R4 and l ' nitrogen atom to which they are united a pyrrolidine radical, 2-, or 3- (lower alkyl) pyrrolidine, piperidine, 2-, 3- or 4- (lower alkyl piperidine, 2,6-di (lower alkyl) piperidine, piperazine, piperazine-4-substituted (of which the substituent is an alkyl or carbalkoxy radical each having 1 to 8 carbon atoms, phenyl, methylphenyl, methoxyphenyl, halophenyl, nitrophenyl or benzyl), azepine, 2-, 3-, 4- or 5- (lower alkyl) azepine, morpholine, thiomorpholine, thiomorpholine-1-  <EMI ID = 153.1>  <EMI ID = 154.1>  <EMI ID = 155.1> a pyrrolidine radical, 2- or 3- (lower alkyl) pyrrolidine, piperidine, 2-, 3- or 4 - inférieur lower alkyl) piperidine, 2,6-di (lower alkyl) piperidine, piperazine, piperazine 4- substituted (the substituent of which is an alkyl or carbalkoxy radical, each of which has 1 to 8 carbon atoms, phenyl, methylphenyl, methoxyphenyl, halophenyl, nitrophenyl or benzyl), azepine, 2-, 3-, 4- or 5- (lower alkyl ) azepine, morpholine, thiomorpholine, thiomorpholine-1-oxide or thiomorpholine-1,1-dioxide, R represents a Cl-18 alkyl radical other than  <EMI ID = 156.1> aryl or lower aralkyl each of which has up to 12 carbon atoms or a heteroalicyclic or heteroaromatic radical of 3 to 8 members, at least two of which are carbon atoms,  <EMI ID = 157.1> R and R independently represent hydrogen atoms or lower alkyl radicals, it being understood that each of the lower alkyl, lower alkanoyl and lower alkoxy radicals mentioned above has 1 to 6 carbon atoms. 2 - Compound according to claim 1, in the dual formula A and B each represent indeoendam- the amidino radical of formula  <EMI ID = 158.1> 3 - Compound according to claim 2, in  <EMI ID = 159.1> of hydrogen and R and R4 each represent a methyl radical. 4 - Compound according to claim 1, which is  <EMI ID = 160.1> methylene-9a-methoxymitosan. 5 - A compound according to claim 1, which is  <EMI ID = 161.1> pylamino) methylene-9a-methoxymitosan. 7 - A compound according to claim 1, which is  <EMI ID = 162.1> no) methylene-9a-methoxy-N -methylmitosan. 8 - Compound according to claim 2, in the formula of which R <1> and R2 each represent a hydrogen atom and R <3> and R4 constitute with the nitrogen atom to which they are united the piperidine radical. 9 - A compound according to claim 1, which is  <EMI ID = 163.1> (1-piperidinylmethylene) mitosan. 11 - Compound according to claim 2, in the  <EMI ID = 164.1> of hydrogen and R and R4 constitute with the nitrogen atom to which they are united the radical morpholine. 12 - Compound according to claim 1 which is <EMI ID = 165.1> (1-morpholino-methylene) mitosan. 13 - Compound according to claim 1, in  <EMI ID = 166.1>  <EMI ID = 167.1> represents the amidino radical of formula  <EMI ID = 168.1> 14 - Compound according to claim 13, in  <EMI ID = 169.1> methyl dical. 15. The compound according to claim 1, which is 7-amino-N-dimethylaminomethylene-9a-methoxyN -methylmitosan. 16 - Compound according to claim 1, which is 7-amino-N -dimethylaminomethylene-9a-methoxymitosan. 17 - Compound according to claim 1, in the formula of which A represents a methoxy radical and B represents the amidino radical of formula  <EMI ID = 170.1> 18 - Compound according to claim 17, in the formula of which R3 and R4 each represent a methyl radical. 19 - A compound according to claim 1, which  <EMI ID = 171.1> formylmitosan. 21 - Compound according to claim 1, in the formula of which A represents one of the amidino radicals indicated and B represents an amino radical. 22 - Compound according to claim 21, in the formula of which A represents the amidino radical of formula  <EMI ID = 172.1>  <EMI ID = 173.1> and, R and R each represents a methyl radical. 23 - Compound according to claim 1, which is 7- (dimethylaminomethylene) amino-9a-methoxymitosan. 24 - Compound according to claim 1, which is 7- (dimethylaminomethylene) amino-9a-methoxy-Nlamethylmitosane. 25 - Process for the preparation of a compound according to claim 1, characterized in that a compound chosen from mitomycin C,  <EMI ID = 174.1> either with an amide-acetal of formula  <EMI ID = 175.1> where R, 2 R <3> and R 4 are as defined in claim 1 and the R8 each independently represent a lower alkyl or cycloalkyl radical having up to 6 carbon atoms or else together constitute an alkylene radical forming with the atoms nitrogen attached and the intermediate carbon atom a 5- or 6-membered ring structure, in solution in an anhydrous liquid organic reaction medium compatible with the reaction at a temperature of 40 to 65 [deg.] C until formation of a reaction product in which B or A and B in the formula of claim 1 represent the amidino radical of formula  <EMI ID = 176.1> 26 - Process according to claim 25, characterized in that the liquid organic reaction medium comprises a lower halogenated aliphatic hydrocarbon and more than two molecular proportions of the amide-acetal relative to mitomycin C are used to obtain a compound in which A and B each represent this amidino radical. 27 - Process according to claim 26, charac-terized in that the reaction medium is chloroform. 28 - Process according to claim 26, characterized in that the reaction medium is a mixture of a lower halogenated aliphatic hydrocarbon and a lower alkanol. 29 - Process according to claim 28, characterized in that the reaction medium is a mixture of chloroform and methanol. 30 - Process for preparing a 7- (substituted amino) -9a-methoxymitosan, characterized in that a compound is reacted according to claim 1 in the formula of which B or A and B represent the radical  <EMI ID = 177.1> amidino of formula R R N-C = N- with an aliphatic, alicyclic, aromatic, heteroaromatic or heteroalicyclic primary amine in which the carbon atom carrying the amino radical carries at least one hydrogen atom and less than two aryl radicals in an anhydrous liquid organic reaction medium compatible with the reaction at a temperature of about -15 [deg.] C to +50 [deg.] C. 31 - Method, characterized in that a first compound according to claim 1 is reacted in the formula of which A and B each represent the radical formula amidino  <EMI ID = 178.1> with more than one proportion molecular tion of an amine selected from aminodiphenylmethane, trifluoroethylamine and t-butylamine at a temperature of 20 to 60 [deg.] C until conversion of the first compound according to claim 1 into a second compound according to claim 1 in the formula of which A represents this amidino radical and B represents an amino radical. 32 - Process according to claim 30 or 31, characterized in that an anhydrous liquid organic reaction medium compatible with the reaction which uses methanol, chloroform, methylene chloride or another lower haloalkane is used. 33 - Process for the preparation of a compound according to claim 1, in the formula of which B represents  <EMI ID = 179.1> lower- 2 (1H) -pyridinylidene) amino or a radical of formula:  <EMI ID = 180.1>  <EMI ID = 181.1> in claim 1, characterized in that a solution, in dimethylformamide (or another compatible solvent), of mitomycin C is reacted with 1.0 at 1.5 molecular proportions of sodium hydride to produce an anionic form of mitomycin C and this anionic form is reacted with an electrophilic reagent capable of forming this amidino radical and chosen between an iminoether, an iminothioether, a halide of halogenethyleneiminium and a saline iminohalide. 34 - A compound according to claim 1, which is 7- (1-methyl-2 (H) -pyridinylidene) amino-9a-methoxymitosan. 35 - A compound according to claim 1 which  <EMI ID = 182.1> tosane. 36 - A compound according to claim 1, which is 7- (1-pyrrolidinylmethylene) amino-9a-methoxymitosan. 37 - A method for inhibiting a tumor in a mammal, characterized in that systemically administered to the mammal carrying a tumor a compound according to claim 1 in an effective anti-tumor dose substantially non-toxic. 38 - 7- [2-Benzylthioethyl] amino-9a-methoxymitosan. 39 - A compound according to claim 1 which  <EMI ID = 183.1> methylamino) ethylidene] -9a-methoxymitosan. 40 - A compound according to claim 1 which  <EMI ID = 184.1> mitosan. 41 - A compound according to claim 1, which  <EMI ID = 185.1> xymitosan. 43 - A compound according to claim 1, which is 7 - [(methoxyamino) methylene] amino-9a-methoxymitosan. 44 - A compound according to claim 1, which is 7 - [(benzyloxyamino) methylene] amino-9a-methoxymitosan. 45 - A compound according to claim 1, which is 7- (1,3-dimethyl-2-imidazolidylidene) -9a-methoxymitosan. 46 - Compound according to claim 1, which is 7- (1,3-dimethyltetrahydropyrimidinylidene) -amino9a-methoxymitosan. 47 - A compound according to claim 1, which is 7- (tetramethyldiaminomethylene) amino-9a-methoxymitosan. 48 - A compound according to claim 1, which is 7- (1-piperidinylmethylene) amino-9a-methoxymitosan.  <EMI ID = 186.1>    <EMI ID = 187.1>  should read:  <EMI ID = 188.1> methyl-amino) ethylidene] -9a-methoxymitosan. - Page 47, line 10 7- (1,3-Dimethyl-2-imidazolinylidene) -9amethoxymitosan. should read: 7- (1,3-Dimethyl-2-imidazolidinylidene) amino-9amethoxymitosan. - Page 50, the formula  <EMI ID = 189.1>  should read:  <EMI ID = 190.1>    <EMI ID = 191.1>  The undersigned is aware that no document attached to the file of an invention patent can be such as to bring, either to the description or to the drawings, modifications substantive and declares that the content of this note does not make such modifications and has no other object than to report one or more material errors. It recognizes that the content of this note cannot have the effect of rendering fully or partially valid the patent which will be granted on the aforementioned patent application, if it was not granted in whole or in part under the law currently in force. It authorizes the Administration to attach this note to the patent file and to issue a photocopy. resulting tion at 30-35 [deg.] for 18 hours. The CCM (20% methanol in methylene chloride) reveals a major major zone and a minor slower purple zone. The solution is concentrated under reduced pressure and the resulting syrup is purified by rapid chromatography to obtain the desired compound which is a dark green amorphous solid (75 mg, 10%). An analytical sample is obtained by precipitation from a solution in methylene chloride using n-hexane.  <EMI ID = 192.1> (m, 10H), 4.02 (dd, 1H, J = 10 and 4Hz), 4.40 (d, 1H, J = 12Hz), 5.06 (t, 1H, J = 10Hz), 5.46 (dd, 1H, J = 10 and 4Hz), 7.90 (s, 1H)  <EMI ID = 193.1> 1520, 1230, 1050 UV (MeOH) &#65533; : 384 and 234 max  <EMI ID = 194.1> calculated C, 55.64; H, 5.80; N, 16.23% found C, 55.07; H, 5.55; N, 15.88% EXAMPLE 19 - 7- (1-Pyrrolidinylmethylene) amino-9a-methoxymitosan  <EMI ID = 195.1>    <EMI ID = 196.1>  A 0.5 molar solution of pyrrolidinylchloromethyleniminium chloride is prepared by adding dropwise oxalyl chloride (3.17 g, 25 millimoles) at 0 [deg.] to a solution of 1-formylpyrrolidine (2.48 g, 25 millimoles) in 50 ml of chloroform and stirring the mixture for 30 minutes at room temperature. Sodium hydride (24 mg, 1 millimole) is added separately in a nitrogen atmosphere to a solution of mitomycin C (334 mg, 1 millimole) in 3 ml of 1-formylpyrrolidine. After 20 minutes of stirring at room temperature, the solution is cooled to about -40 at -50 [deg.] and the solution of the iminium salt prepared above (1 ml, 0.5 millimole) is added thereto. We add to this mixture alternately, at 10 minute intervals, 12 mg (0.5 millimole) NaH, 0.5 ml (0.25 millimole) of the iminium salt solution, 6 mg (0.25 millimole) of NaH, 0.25 ml (0.125 millimole) of the iminium salt solution and finally 3 mg (0.125 millimole) of NaH and 0.125 ml ( 0.063 millimole) of the iminium salt solution. After 30 minutes of stirring at -30 [deg.], The mixture is warmed up to room temperature. It is diluted with ethyl acetate and the inorganic salt is filtered off. The residue from the evaporation of the solvent is subjected to TLC on silica gel (10% methanol in methylene chloride). By extraction of the green band, 120 mg (15 /) of the desired compound are obtained.  <EMI ID = 197.1> <EMI ID = 198.1> 4.03 (dd, 1H, J = 10.4 Hz), 4.44 (d, 1H, J = 12 Hz), 5.05 (t, 1H, J = 10 Hz), 5.45 (dd, 1H, J = 10, 4 Hz), 8.04 (s, 1H).  <EMI ID = 199.1> 1560, 1300, 1055. EXAMPLE 20 - 7- [N-Methyl-N- (methylamino) methyl] amino-9a-methoxymitosan  <EMI ID = 200.1> The procedure of Example 17 is repeated, replacing the mitomycin C by an equal molecular quantity of 9a-methoxy-7- (N-methylamino) mitosan (Matsui et al., The Journal of Antibiotics, XXI, 189- 198 (1968)). EXAMPLE 21 - Compound XXIII  <EMI ID = 201.1> amino) ethylidene] -9-methoxymitosan  <EMI ID = 202.1>  A suspension of 600 mg of mitomycin C (1.79 millimole) in 2 ml of methanol is prepared and 3 ml of N, N-dimethylacetamide dimethylacetal are added thereto. The suspension is heated to 75-80 [deg.] For 2 hours with stirring. At this time, TLC (methylene chloride / methanol 10: 1) reveals that almost all of the mitomycin C has been consumed by the. reaction. The product appears as a green area.  The solvent and volatile constituents are removed by evaporation of the reaction mixture to dryness under reduced pressure to obtain a residue which is dissolved in methylene chloride and which is poured onto a column of silica gel (40 g) which developing with 1% methanol in methylene chloride (200 ml), 2% methanol in methylene chloride (200 ml) and 5% methanol in methylene chloride (400 ml). We combine the fractions containing the green zone where the product is found and they are concentrated into an amorphous solid with a weight of 110 mg (yield of 13%). This solid is dissolved in 2 ml of acetone and reprecipitated therefrom by addition of hexane. The product is collected by filtration.  <EMI ID = 203.1> calculated C, 58.46; H, 6.83; N, 17.79% found C, 58.89; H, 6.89; N, 17.64%  <EMI ID = 204.1> 1620, 1580, 1550, 1300, 1055.  <EMI ID = 205.1> compatible with the structure of the desired compound. EXAMPLE 22 Compound XXIV 7- [1- (Dimethylamino) ethylideneamino] -9a-methoxymitosan  <EMI ID = 206.1> A solution of 100 mg (0.21 millimole) of compound XXIII in 2 ml of chloroform is added to 2 ml of aminodiphenylmethane and the solution is heated for a few hours at 55-60 [deg.]. At this time, traces of compound XXIII remain in the reaction mixture which is nevertheless concentrated to a residue which is chromatographed on neutral alumina which is eluted with a gradient starting with methylene chloride and ending with methanol / methylene chloride 2.5: 1. The main green zone is isolated in the form of a green amorphous solid with a weight of 25 mg (29.4%). The compound is purified by dissolution in acetone and adding hexane to the acetone solution until precipitation occurs. The product is collected by filtration and dried.  <EMI ID = 207.1> calculated C, 56.58; H, 6.20; N, 17.37% found C, 55.71; H, 6.34; N, 15.23%  <EMI ID = 208.1> compatible with the structure.