CH667094A5 - SUBSTITUTED 7-OXOMITOSANS. - Google Patents

Description

DESCRIPTION

The present invention relates to novel mitosan analogs containing a disulfide group and processes for their preparation. These compounds are mitomycin A analogs, in which the 7-alkoxy group carries an organic substituent which includes a disulfide group. The present invention also provides a process for the preparation of mitomycin A and derivatives thereof. Mitomycin A is an antibiotic of recognized utility and the 7-0 substituted mitosan analogs thereof have a similar purpose.

Nomenclature: The systematic name according to "Chemical Abstract" for mitomycin A based on the recent revision is (Shirhata et al., "J. Am. Chem. Soc.", 105, 7199 [1983]):

[laS- (lass, 8ß, 8aa, 8bß)] - 8 - [((aminocarbonyl) oxy) methyl] -. 6,8a-dimethoxy-1,1 a, 2,8,8a, 8b-hexahydro-5-methylarizino [2 ', 3', 3,4] pyrrolo [1,2, a-indole-4,7 -dione after which the azirinopyrroloindole ring system is numbered as follows:

wherein R1, R3, R4, Alki and Alk2 have the meaning given in claim 1 and Ar is the organic radical of a diazotizable, aromatic amine, is reacted under the reaction conditions of an inert, organic solvent at 0-60 ° C. 65

23. The method according to claim 22, characterized in that the triazene of the formula VI is 1- [2- (3-nitro-2-pyrodyldi-thio) ethyl] -3- (4-methylphenyl) triazene.

"Chemical Abstracts"

667 094

4th

A trivial nomenclature system, which has found widespread use in Mitomy literature, identifies the above ring system including some of the characteristic substituents of Mitomycins as Mitosan.

10th

According to this system, mitomycin A is 7,9a-dimethoxy-mitosan and mitomycin C is 7-amino-9a-methoxymitosan. As for the stereochemical configuration of the products of the present invention, it is intended that when drawn by the tribe name "Mitosan" or by the structural formula, their stereochemical configuration is the same as that of Mitomycin A or C.

Mitomycin C is a fermentatively produced antibiotic and is currently on sale with the approval of the Food and Drug Administration for the therapy of disseminated adenocarcinoma of the stomach or pancreas, in tested combinations with other approved chemotherapeutic agents and as a soothing agent if other modalities are unsuccessful (Mutamycin®, Bristol Laboratories, Syracuse, New York 13221, Physicians' Desk Reference, 37th Edition, 1983, pp. 747-748). Mitomycin C and its production by fermentation is the subject of U.S. Patent 3,660,578, issued May 2, 1972, claiming priority from earlier applications including an application filed in Japan on April 6, 1957.

The structures of Mitomycin A, B, C and Porfiro-mycin were first described by J.S. Webb et al. from the department Lederle Laboratories Dicision American Cyanamid Company in «J. At the. Chem. Soc. », 84.3185-3187 (1962). One of the chemical conversions used in this structural study in connection with mitomycin A and mitomycin C was the conversion of the former, 7,9a-dimethoxymitosan, by reaction with ammonia to the latter, 7-amino-9a-methoxymitosan. The displacement of the 7-methoxy group of mitomycin A has proven to be a reaction of considerable interest in the preparation of the antitumor active derivative of mitomycin C. Recently, the stereochemical configurations of positions 1, la, 8a and 8b are as described above according to the «Chemical Abstracts »nomenclature have been shown (Shirhata et al.,« J. Am. Chem. Soc. », 105, 7199-7200 [1983]). The previous literature relates to the enantiomer.

The following articles and patents relate, inter alia, to the conversion of mitomycin A to a 7-substituted aminomitomycin C derivative which has anti-tumor activity. The object of this research was to produce derivatives that are more active and, in particular, less toxic than Mitomycin C:

Matsui et al., "J. Antibiotics », XXI, 189-198 (1968);

Konishita et al., "J. Med. Chem. », 14.103-109 (1971);

Iyengar et al., "J. Med. Chem. », 24.975-981 (1981); io Iyengar, Sami, Remers and Bradner "Abstracts of Papers", 183rd Annual Meeting of the American Chemical Society, Las Vegas, Nevada, March 1982, "Abstract No. MEDI 72";

Cosulich et al., U.S. Patent 3,332,944, issued July 25, 1967; i5 Matsui et al., U.S. Patent 3,420,846, issued January 7, 1969;

Matsui et al., U.S. Patent 3,450,705, issued June 17, 1969;

Matsui et al., U.S. Patent No. 3,514,452, issued May 26, 1970;

Nakano et al., U.S. Patent 4,231,936, issued November 4, 1980;

20 Remers, U.S. Patent 4,268,676, issued May 19, 1981.

The following patent applications relate to the preparation of 7-substituted aminomytin-C derivatives in which the substituent contains a disulfide bond.

Kono et al., European Patent Application No. 116 208 25 (1984);

Vyas et al., GB-A-2 140 799 (1984).

The 7-alkoxy-substituted mitosanes, which are structurally related to mitomycin A, according to an article by Urakawa et al., «J. Antibiotics », 23, pp. 804-809 (1980) represent 30 useful antibiotics which have been shown to be active in experiments with experimental animal tumors.

Mitomcyin C is the major mitomycin produced by fermentation and is the commercially available form. The common technique for converting 35 mitomycin C to mitomycin A has a number of shortcomings. The hydrolysis of mitomycin C in the corresponding 7-hydroxy-9a-methoxy-mitosan and the subsequent methylation of this substance requires dia-zomethane, which is a very dangerous substance when handled on an industrial scale, and the 7-hydroxy intermediate is very unstable (Matsui et al., "J. Antibiotics", XXI, 189-198 [1968]). One attempt to avoid these difficulties involves the use of 7-acyloxymitosans (Kyowa Hakko Kogyo KK, JP-PS J5 45 6073-085, Farmdoc No. 56227 D / 31). The alcoholysis of mitomycin A as described by Urakawa et al. in «J. Antibiotics », 23, pp. 804-809 (1980), is only limited to the production of those specific 7-alkoxy structure types whose alcohol starting material has been found to be available and reactive.

The present invention relates to the mitomy cin A analogs of the formula I.

wherein R2 is an organic group, namely the structural component of an organic thiol of the formula R2SH, where R2 is R3-alkali or R4. Alternatively, these compounds can also be defined by formulas II and III according to claim 1. The meanings of

5

667 094

Symbols R1, R3, R4, Alki and Alk2 are specified in claim 1.

The compounds according to the invention are inhibitors of experimental animal tumors. In particular, the substances of Examples 17, 20 and 21-34 are new compounds. They are used in a similar way to Mitomycin C. The doses used are adjusted according to the ratio of their toxicity to the toxicity of mitomycin C. In cases where the new compound is less toxic, a higher dose is used.

Another object of the present invention is the new process for the preparation of the mitosanes of the formulas II and III. In this process, a mitosan of the formula

CH.

I-R-

with a triazene of formula V or VI

Ar-N = N-NH-Alkî-SS-Alki-R3 V

Ar-N = x N-NH-Alkî-SS-R4 VI

implemented, wherein R1, R3, R4, Alki and Alk2 are as defined above and Ar is the organic radical of a diazotizable aromatic amine.

The invention also relates to the following process for the preparation of the mitosanes of the formulas II and III. This process involves reacting a thiol of formula VII or VIII

R3AlkiSH R4SH VII VIII

with a mitosan derivative of the formula

NO.

OCH

(Ib)

i-R '

CH

\

The disulfide mitosanes of the formula Ib are prepared by the triazene process described here. Specifically, the preparation of the mitosan of Formula Ib, wherein Alka is ethylene and R1 is hydrogen, is described in Example 20 and in parallel U.S. Application No. 646,888, filed September 4, 1984.

The terms "lower alkyl", "lower alkoxy" and "lower alkanoyl" used in the description and the claims mean (unless expressly stated otherwise in the appropriate place) straight or branched chain alkyl, alkoxy or alkanoyl with 1-6 C atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, amyl, hexyl, etc. These groups preferably contain 1-4 C atoms and particularly preferably 1-2 C atoms. Unless otherwise stated in the appropriate place, the term used in the description and claims means

«Halogen» chlorine, fluorine, bromine and iodine. The term "non-toxic, pharmaceutically acceptable salt" means salts of the compounds of formulas I and II with any non-toxic, pharmaceutically acceptable acid or base. Such acids are well known and include hydrochloric acid, hydrobromic acid, sulfuric acid, benzoic acid, methanesulfonic acid, tartaric acid, citric acid, camphorsulfonic acid, levulinic acid and the like. Such bases are well known and include e.g. non-toxic metallic salts such as sodium, potassium, calcium, magnesium and ammonium salts and salts with non-toxic amines, e.g. Trialkyla-mine, procaine, dibenzylamine, pyridine, N-methylmorpholine and N-methylpiperidine. The salts are produced by known processes.

The new process of the present invention, which uses l-substituted-3-aryltriazenes, can also be used to prepare compounds of formulas II or III by using a mitosane of formula IV with a triazene of formula V or VI, as in the scheme below

2, is implemented.

I-R "

CH

IV

Ar-N = N-NH-Alk, -SS-Alk, -R3

V 1

+> II or III

or Ar-N = N-NH-Alk2-SS-R4

VI

wherein R1, R3, R4, Alki and Alk2 are as defined above and Ar is the organic radical of a diazotizable aromatic amine.

Aryltriazines of the formulas V or VI can be prepared in a similar manner to that described above for the preparation of the aryltriazenes of the formula VI, the alkylamines used being aminodisulfides of the formula XII

R2-SS-Alk2NH2 (XII)

which are alternatively described by formulas XIII and XIV

R3-Alk.-SS-Alk2NH2 and R4-SS-Alk2NH2

XIII XIV

Aminodisulfides of the formulas XIII and XIV are known compounds and can be prepared by various methods. For example, they can be reacted with an appropriate thiol R3AlkiSH or R4SH with a colored salt of the formula XV

NH2Alk2SSO> Na XV)

or with a sulfenylthiocarbonate of the formula

O

II

NH2Alk2SSCOCH3 (XVI)

be preserved.

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

667 094

6

Klayman et al., "J. Org. Chem. », 29, pp. 3737-3738 (1964), have produced the following compounds according to the Bunte salt method:

2-aminoethyl-n-butyl disulfide 2-aminoethyl-n-hexyl disulfide 2-aminoethyl-n-octyl disulfide 2-aminoethyl-n-dexyl disulfide 2-aminoethyl-phenyl disulfide 2-aminoethyl-benzyl disulfide

Methanol has proven to be the preferred reaction solvent for the reaction of the colored salt with the thiol. The reaction temperatures of 0 to 10 ° C were found to be preferred when using this solvent. With other solvents, higher temperatures were necessary. The main disadvantage of this process is the formation of the symmetrical disulfide as a by-product, presumably as a result of disproportionation of the desired mixed disulfide.

The mixed disulfide starting materials of the formulas XIII and XIV are preferably prepared via the reaction of a suitable thiol with a sulfenylthiocarbonate of the formula XVI. This is the method of S. J. Brois et al., «J. At the. Chem. Soc. », 92, 769-7631 (1970). Typically, this manufacturing process involves adding the thiol to a methanol solution of amino-alkylsulfenylthiocarbonate of Formula XVI and proceeding the reaction at a temperature in the range of 0-25 ° C. The reaction times vary from essentially instantaneously to a few hours, depending on the particular thiol used. The progress of the reaction can be followed by determining the presence of the unreacted thiol in the reaction vessel. If the reaction is sluggish, a catalytic portion of triethylamine can be added as a reaction accelerator.

The 1- (substituted disulfide) -3-aryltriazenes of the formulas V or VI are obtained by reacting aminodisulfides of the formula XII with aryldiazonium salts in a manner similar to that described above for the preparation of aryltriazenes of the formula VI. Any arylamine with 6-12 C atoms, which easily forms a diazonium salt, can be used as a source for the aryl part of the 1,3-disubstituted triazene. Some examples of the disulfide-triazenes prepared in this way and used in the present invention are 1- (2-acetamidoethyldithio) ethyl) -3- (4-methylphenyl) triazene;

1- (2- (3-Nitro-2-pyridyldithio) ethyl) -3- (4-methylphenyl) triazene.

Below are some useful triazene starting materials of formulas V and VI for use in the present invention.

1- (2- (3-nitro-2-pyridyldithio) ethyl) -3- (4-chlorophenyl) triazene;

1- (2- (3-nitro-2-pyridyldithio) propyl) -3- (4-methylphenyl) triazene;

1 - (2- (2-pyridyldithio) ethyl) -3- (4-methylphenyl) triazene; 1 - (2- (phenyldithio) ethyl) -3- (4-methylphenyl) triazene; 1 - (2- (butyldithio) ethyl) -3- (4-methylphenyl) triazene; 1- (2- (4-methoxyphenyldithio) ethyl) -3- (4-methylphenyl) triazene;

1- (2- (4-nitrophenyldithio) ethyl) -3- (4-methylphenyl) triazene;

1- (2 - ((2-Benzoylaminoethyl) dithio) ethyl) -3- (4-methylphenyl) triazene;

1- (2- (4-chloro-2-naphthyldithio) ethyl) -3- (4-methylphenyl) triazene;

1- (2-cyclopropylmethyldithio) ethyl) -3- (4-methylphenyl) triazene;

1- (2 - ((2-Phenoxyethyl) dithio) ethyl) -3- (4-methylphenyl) triazene.

In a preferred embodiment of the present invention, a further process for the preparation of the disulfide mitosanes of the formula Ia is proposed in which the organic group R2 is derived from an organic thiol of the formula R2SH, where R2 has the meaning 20 R3Alki or R4, where R3, R4 and Alki have the meaning given above.

For the preparation of the disulfide mitosanes of the formula Ia, 9a-methoxy-7- (2- (3-nitro-2-pyridyldithio) -ethoxy) mitosane of the formula XVII is preferably used, a thiol-25 exchange process having a suitable organic thiol of the formula R2SH is carried out according to reaction scheme 3. The driving force behind the formation of the disulphides of formula Ia is the stability of the by-product, namely 3-nitro-2-mercaptopyridine, which exclusively exists from thion XVIII.

Scheme 3

+ R SH>

50

O

R2-SS-CH2CH2Q

55

Yes

XVIII

Alternatively, if the production of mitosans of formulas II or III, in which Alla is different from ethylene, such as trimethylethylene or propylene, is desired, a convenient triazene of formula V or VI can be used in a process as shown in Scheme 2 , whereby a disulfide mitosan of formula Ib is obtained,

MO.

(Ib)

och

CH

I-R '

wherein Alk2 and R1 are as defined above.

Two synthesis methods for the production of lipophilic as well as hydrophilic mitosanes of the formula Ia are specified here. Process A is used for the preparation of either lipophilic or moderately soluble disulphides of the formula Ia, while process B is used for water-soluble disulphides of the formula Ia, which is preferably isolated as the sodium salt or in zwitterionic form. Preferably at least one equivalent of the mercaptan R2SH is used per equivalent of mitosan of the formula XVII and the reaction can be carried out in the presence of one equivalent of base per equivalent of mercaptan R2SH. Preferred bases are the tertiary amines, e.g. Triethylamine, N-methylmorpholine, N-methylpiperidine, pyridine, 2,6-lutidine and the inorganic bases, e.g. Sodium bicarbonate, potassium carbonate or potassium bicarbonate. Suitable inert solvents for the reaction of the starting material of the formula XVII and R2SH are the lower alkanols, the lower alkyl esters of lower alkane carboxylic acids, lower aliphatic ketones, cyclic aliphatic ethers, lower polyhalogenated aliphatic hydrocarbons and water. The organic solvents contain up to 8 carbon atoms, but these are preferred with boiling temperatures of less than 100 ° C. Specific preferred solvents are methylene chloride, methanol, acetone, water and mixtures thereof. The reaction can be carried out at the reflux temperature of the reaction mixture or up to 60 ° C. It is preferred to carry out the reaction at room temperature or below, e.g. in the range of 0-25 ° C.

The reaction conditions and precautionary measures described above can be used for the preparation of other disulfide mitosanes of the formulas Ia and Ib according to the general process shown in Scheme 3.

Below is a list of representative thiols of the formula R3AlkiSH or R4SH, which can be converted via the reaction with the colored salt XV or the sulfenylthiocarbonate XVI to produce the intermediates of the formulas XIII and XIV, which can then be converted into the products according to the invention. In the preferred embodiment, the representative thiols can be used in a reaction with mitosanes of the formula Ia or Ib to prepare the compounds according to the invention. The only limitations in the methodology of the present invention are the use of thiols containing terminal primary alkyl amines which can result in a mixture of products and the use of heteroaromatic thiols which cannot react with the compounds of formula Ia or Ib .

667 094

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hsch2ch3

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667 094

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r = h, ch3

n = 1-3; m = 1-3

r2 = h, ch .; r1 - h, ch3

r1 = h, ch3

r = h, ch3

n (ch3) 2

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667 094

12

The usefulness of the compounds of formula I for anioneoplastic therapeutic methods is demonstrated by the in vivo screening methods, in which the compounds were administered in varying dose proportions to mice which were induced with P-388 leukemia or a B16 melanoma was.

It is believed that the compounds of the present invention have antibacterial activity against gram-positive and gram-negative microorganisms in a manner similar to that observed for the naturally occurring mitomycins; the compounds of the invention may therefore be useful as therapeutic agents for the treatment of bacterial infections in humans and animals.

Activity against murine leukemia P-388

Table I contains the results of laboratory tests with CDFi mice which were implanted intraperitoneally with a tumor inoculum of 106 ascites cells of murine leukemia P-388 and which were treated with different doses of either a test compound of the formula I or II or with mitomycin C. The compounds were administered by intraperitoneal injection. Groups of 6 mice were used for each dose level and treated with a single dose of the compound the day after the inoculation. A group of 10 saline control mice were included in each series of experiments. In a 30-day protocol, the mean survival in days determined for each group of mice and the number of survivors at the end of the 30-day period were recorded. The mice were weighed before each treatment and again on day 6. The weight 5 change was taken as a measure of the toxicity of the drug. Mice weighing 20 g each were used and a weight loss of about 2 g was not considered excessive. The results were determined as% T / C, which means the ratio of the mean survival time of the treated group to the mean survival time of the control group treated with saline x 100. The control animals treated with saline normally died within 9 days. The “maximum effect” in the following table is expressed as% T / C and by 15 the dose by which this effect was achieved. The values in parentheses are those obtained with mitomycin C as a positive control in the same experiment. As a result, the extent of the relative activity of the substances present to mitomycin C can be estimated. A minimal effect, determined as% T / C, was considered 125. The minimum effective dose shown in the table below is that which gives a% T / C of 125. The two values, which are given in the "mean weight change" column, are the mean weight change per mouse at the maximum effective dose or the minimum effective dose.

Table I Inhibition of Murine Leukemia P-388

Connection of example no.

% t / c maximum effect dose minimal effect

^ • 1 dose of mean weight change2

10th

-ch2ch2ss

NH ". 194 (> 333) 3.2 (4.8}

<0.1

-3.1; +1.8

17th

-ch2ch2ssch2 ch3conhch2

183 (306)

6.4 (3.2)

0.1

-1.2; -0.3

20th

N02

-ch2CH2SS- ^ y

206 (> 333)

6.4 (4.8)

■ 0.2

-1.0; + 1.S

26

-ch2ch2ss - <ryno2

178 (> 333)

12.8 (4.8)

0.2

+0.4; +2.1

1. mg / kg body weight

2. mean value in g per day for each minimum and maximum effective dose

3. Urakawa et al., "J. Antibiotics", 23, 804-809 (1980)

4. Values in brackets refer to tests with mitomycin C in the same experiment

13

667 094

Activity against melanoma B16

Table II contains the results of antitumor tests using melanoma B16 grown in mice. BDFi mice were used and inoculated subcutaneously with the tumor implant. A 60 day protocol was used. Groups of 10 mice were used and the mean survival for each group was determined. In the same manner as the test animals, control animals were inoculated and treated with the injection vehicle, and an average survival of 24 hours was achieved without drugs. The survival time based on that of the controls (% T / C) was used as a measure of the effectiveness and the maximum effective dose and the minimum effective dose were determined for each test compound. The minimum effective dose was defined as that which gave a% T / C of 125. For each dose level, the test animals were treated intravenously with the test compound on day 1, 5 and 9.

Table II B16 melanoma

Connection Maximum effect

Minimal

Middle one

of the example% T / C dose1

effective

Weight

No.

Dose1

modification

10 167 (112) 3 1.6 (3) 3

<0.4

-0.9

+ 1.4

> 214 (145) 2.4 (3)

<1.6

-2.4

-1.9

2 110 (112) 3.2 (3)

3.2

+ 0.5

+ 0.5

8 152 (145) 1.6 (3)

<1.6

-0.6

-0.6

1 mg / kg body weight

2 mean value in g per day for each maximum and minimum effective dose

3 Values in brackets refer to mitomycin C tests in the same experiment

In view of the anti-tumor activity, which was observed in the experimental animal tumor, the compounds according to the invention can be used for the inhibition of tumors in humans and mammals. For this purpose, they are administered systemically and in a substantially non-toxic, anti-tumor dose to the tumor-bearing human or animal.

The compounds according to the invention are used primarily for administration by injection, predominantly in the same way and for the same purpose as mitomycin C. Depending on the particular sensitivity to tumors, somewhat smaller or larger doses can be used. They can easily be commercialized as dry pharmaceutical compositions containing diluents, buffers, stabilizers, solubilizers and / or additives to impart pharmaceutical elegance. These compositions can then be assembled with an injectable liquid medium immediately before use. Useful injectable liquids include water, isotonic saline and the like.

In the following procedures and examples, all temperatures are given in ° C and the melting points are uncorrected. The magnetic proton resonance spectra ('H NMR) were carried out on a Varian XL100, Joel FX-90Q or Bruker WM360 spectrometer either in pyridine-ds or in d2o as indicated. When pyridine-d5 was used as the solvent, the pyridine resonance at 5 = 8.57 served as the internal reference, while when using d2o as the solvent, TSP served as the internal reference. The chemical shifts are given in 8 units and the coupling constants in Hertz. The splits are given as follows: s single «, d doublet, t triplet, q quartet, m multiple«, bs broad signal, dd doublet of a doublet, dt doublet of a triplet. The infrared spectra were determined either on a Beckmann model 4240 spectrometer or a Nicolet-5-DX-FT-IR spectrometer and are given in reciprocal centimeters. The ultraviolet (UV) spectra were carried out either on a Cary spectrometer, model 290, or on a Hewlett-Packard 8450A spectrometer equipped with a multi-diode directional detector. Thin layer chromatography (TLC) was carried out on 0.25 mm Analtech silica gel GF plates. Flash chromatography was carried out either on neutral aluminum oxide, Woelm (DCC grade) or silica gel, Woelm (32-63 µm) and with the solvents indicated. All evaporation of solvents was carried out under reduced pressure and below 400 ° C.

The l-alkyl-3-aryltriazenes are a class of reagents which are known to react with carboxylic acids and form the corresponding lower alkyl esters. l-Methyl-3- (4-methylphenyl) triazene can be prepared as follows:

Procedure 1

E.H. White et al., «Org. Syn. », 48, pp. 102-195 (1968).

l-methyl-3-p-tolyltriazene. p-Toluidine (50.2 g, 0.47 mol) is placed in a 2-1 bottle equipped with a 200 ml dropping funnel and an efficient stirrer, and the bottle is placed in an ice salt bath from -10 ° C hung. A solution of 46.8 g (0.55 mol) of potassium nitrite in 150 ml of water is added to the dropping funnel, and a mixture of 250 g of chopped ice and 140 ml of concentrated hydrochloric acid is added to p-toluidine with stirring. The potassium nitrite solution is slowly added with continued stirring for 1-2 hours until a positive starch potassium iodide test is noted (Note 1) and the mixture is additionally stirred for 1 hour to ensure that all of the toluidine is reacted.

The solution of p-toluenediazonium chloride is then brought to a pH of 6.8-7.2 at 0 ° C. with cold concentrated aqueous potassium carbonate, the solution becoming red to orange and a small proportion of red material precipitating out. The cold neutral solution is transferred to a dropping funnel and slowly added to a vigorously stirred mixture of 150 g of sodium carbonate, 300 ml of 30-35% aqueous methylamine (note 2) and 100 g of chopped ice in a 3-1 bottle. The reaction mixture is maintained at a temperature of approximately -10 ° C during the addition, which takes approximately 45 minutes (Note 3). The solution is extracted with three 1 liter parts of ether. The ethereal extracts are dried with anhydrous sodium sulfate and evaporated on a rotary evaporator at room temperature to give 65 g of crude l-methyl-3-p-tolyltriazene (Note 4). This is placed in a water-cooled sublimation device and the triazing is sublimed at 50 ° C (1 mm); 43.3 g (0.29 mol, 62%) of a yellow crystalline sublimate F. 77-80 ° C is obtained (Note 5). The sublimate can be recrystallized from hexane, the triazine being obtained as white needles, melting point 80.5-81.5 ° C. Usually it is dissolved in a minimal amount of ether and the solution is diluted with 2 volumes of hexane and cooled to 0 ° C to give weak platelets of a pale yellow excretion; M. 79-81 ° C. The yield of pure triazene is 33-37 g (47-53%) (Note 6).

5

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50

55

60

65

667 094

14

Remarks

1. The individual tests with starch potassium iodide paper should be done 1-2 minutes after the addition of sodium nitrite is complete.

2. Can be replaced with 40% aqueous methylamine.

3. The reaction is complete when a drop of solution with a solution of β-naphthol in the aqueous sodium carbonate no longer produces a red color.

4. The main impurity is 1,5-di-p-tolyl-3-methyl-1,4-pentazadiene (mp 148 ° C). This can be removed by fractional crystallization, but it is easier to sublimate the triazene from the reaction mixture.

5. The sublimate contains a trace of 1,3-di-p-tolyltriazene, as shown by thin layer chromatography. The recrystallization gives the pure l-methyl-3-p-tolyltriazene.

6. This process works only well with water-soluble amines. Method 2 below is more appropriate for the preparation of triazines from water-insoluble amines.

Procedure 2

E.H. Whit et al., "Tetrahedron Letters", No. 21, p. 761 (1961).

l-n-butyl-3-p-chlorophenyl triazene. A solution of p-chlorobenzenediazonium hexafluorophosphate (recrystallized from acetone / methanol, 2.87 g, 10.1 mmol) in dimethylformamide (dimethylamine-free) was slowly added to a stirred mixture of n-butylamine (0.73 g, 10.0 mmol) of powdered sodium carbonate (15 g) and dimethylformamide (30 ml) were added, stirred and kept at -5 ° C. The diazonium salt solution can be used at room temperature; However, a purer product is obtained if the diazo-nium salt solution is prepared in a separating funnel at a temperature of -50 ° C. and is released from such a funnel. The mixture is warmed to 0 ° C and stirred until a negative test is obtained with 2-naphthol (it usually only takes a few minutes). Then ether is added and the mixture is then filtered, and the filtrate is carefully washed with water and dried (the triazene can at this point be recrystallized from pentane at low temperatures).

Procedure 3

7-hydroxy-9a-methoxymitosan. Mitomycin C (2.2 g, 6.6 mmol) was dissolved in 140 ml of 0.1N methanolic NaOH (50%) and the reaction mixture was stirred at room temperature for 30 h. The mixture was then adjusted to pH 4 with IN HCl and extracted with ethyl acetate (4 × 500 ml). The combined ethyl acetate extracts were dried (Na2SC> 4) and evaporated under reduced pressure at 30-35 ° C to give a solid residue which, when diluted with ether and treated with excess hexane, gave a purple precipitate. The precipitate was collected and air dried to give the title compound as a fine purple powder (1.4 g, 63%).

'H NMR (pyridine-ds, 5): 2.05 (s, 3H), 2.14 (bs, 1H), 2.74 (bs, 1H), 3.13 (d, 1H), 3.24 (s, 3H), 3.56 (d, IH), 4.00 (dd, 1H), 4.37 (d, IH), 5.05 (t, IH), 5.40 (dd, IH) , 5.90 (bs, 2H).

Procedure 4

Mitomycin A. An amount of 100 mg (0.30 mmol) of 7-hydroxy-9a-methoxymitosan and 100 mg (0.67 mmol) of 3-methyl-l-p-tolyltriazene was dissolved in 2 ml of methylene chloride and 10 ml of diethyl ether. After 6 hours of moderate reflux, the solution was stirred at room temperature for 18 hours. Thin layer chromatography (methylene chloride: methanol, 90:10) showed the appearance of a dark purple spot at an Rr of 0.36 with a trace of contamination at Rf = 0.41. The reaction mixture was concentrated to dryness and chromatographed on neutral alumina Woelm using methylene chloride and methylene chloride: methanol (30: 1) as eluents. The fractions containing the component with the Rp value of 0.36 were pooled and concentrated to dryness. Precipitation of the dry residue from methylene chloride and hexane gave the title compound as a fine amorphous purple powder.

(25 mg, 24%), mp 161 ° C.

Analysis for C16H19N3O6:

Calculated: C 54.96 H 5.44 N 12.02 Found: C 53.96 H 5.37 N 11.99

IR (KBr), ymax, cm- ': 3400.3300.2950, 1700.1630, 1575, 1200.1060.

'H NMR (pyridine-ds, 5): 1.82 (s, 3H), 2.74 (dd, 1H), 3.12 (d, 1H), 3.24 (s, 3H), 3.54 (dd, IH), 3.96 (dd, IH), 4.02 (s, 3H), 4.22 (d, 1H), 4.84 (bs, 2H), 5.02 (t, IH) , 5.38 (dd, 1H).

The yield in Process 4 can be increased to 63% when methylene chloride is used as the reaction solvent at room temperature for 24 hours.

Procedure 5

Solid NaîCCb, 35% aqueous solution of amine (fraction as in method 1) and ice were placed in a 250 ml round-bottomed flask and the solution was stirred at -5 ° C. (ice salt bath). A cold suspension of p-chlorobenzenediazonium hexafluorophosphate (Aldrich Chemical Co.) in ice, water, NaîCCh (solution approximately pH 7) was added dropwise to this suspension. After the addition was complete, the reaction mixture was extracted with diethyl ether. The combined diethyl ether extracts were washed with water, dried (NaîS04) and concentrated. The yellowish solid residue was purified by column chromatography over Woelm alumina and using hexane / methylene chloride (1: 1) as the eluent ('H NMR excellent).

Triazen production 1

The triazing of Table III below was made according to Method 1 described above. The preparations were purified by column chromatography on Woelm alumina.

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15

Table III

667 094

rnh2 +

preparation

• Ami n

Triazenes

JUST

Diazonium salt triazene ch3ocnhch2ch2ssch2ch2nh2 ch ^ n2 + ci ~ p-ch3c6h4n-m-nhch2ch2ssch2ch2mhcoch3

Triazen production 2

1- (2- (3-Nitro-2-pyridyldithio) ethyl) -3- (4-methylphenyl) triazene

A solution of 4-methylphenyldiazonium chloride is prepared as described in process 1 from p-toluidine and the pH is adjusted to 6.8-7.2 at 0 ° C. as described in this process. A solution containing 21.15 mmol of the diazonium salt in 45 ml of solution was prepared in this way and placed in a dropping funnel which was connected to a 250 ml three-necked round bottom flask, which contained 5.34 g (20.0 mmol ) 2- (3-nitro-2-pyridyldit-hio) ethylamine, 7 g sodium carbonate and 150 ml dioxane, which was added to the bottle in this order. Saturated aqueous sodium carbonate (60 ml) and 10 g ice were added to the bottle. The bottle was cooled in an ice bath and the contents were stirred mechanically. The diazonium salt solution was then dropped from the dropping funnel over a period of 1 hour. After the reaction was completed, the reaction mixture was warmed to room temperature and extracted with three 400 ml portions of ether. Drying and evaporation of these extracts gave the desired product, which was purified by chromatography using an aluminum oxide packed column with a diameter of

2.5 cm and 25 cm in length and using hexane: methylene chloride (4: 1), hexane: methylene chloride (3: 2), hexane: methylene chloride (1: 4) and finally methylenechlo-20 rid containing 1% methanol for the development and elution of the pillar. The appropriate fractions (identified by TLC) were combined and evacuated to give 2.5 g of the title compound.

25 examples

General procedure for the preparation of 7-alkoxy-9a-methoxymitosanes

A solution of triazene (2.4 equivalents) in ch2ck: methanol (4: 1) was added to a solution of 30 7-hydroxy-9a-methoxymitosan (prepared according to method 3) in ch2ck: methanol (4: 1). The reaction mixture was stirred at room temperature and the progress of the reaction was followed by thin layer chromatography (TLC, 10% MeOH in ch2ci2). The 7-alkoxy-9a-35 methoxymitosan product appeared as a dark purple spot on the TLC. The reaction mixture was chromatographed on Woelm alumina when the reaction was thought to be complete based on TLC and the 7-alkoxy-9a-methoxymitosane was obtained as an amorphous solid 40.

Table IV

7-alkoxy-9a-methoxymltosane

Si NMR (Pyridine-dj, 6) IR (cm "" 1)

Example formula ppin ~

No.

R — CHjCH2SS-CH2CH2NHCOCH3 1.88 (8, 3H), 2.04 (s, 3H), 2.24 (ba, 1H), 3300, 2920,

2.76 (bs, 1H), 3.08 (m, SH), 3.24 (s, 3H), 1710, 1625, 3.52 (d, 1H), 3.76 (t, 2H), 3.98 (dd, 1H>, 1210, 1065 4.24 (d, IH), 4.72 (t, 2H), 5.04 (t, 1H),

5.40 (dd, 1H).

667 094

16

Procedure 6

9a-methoxy-7- (2- (3-nitro-2-pyridyldithio) ethoxy) mitosane

7-Hydroxy-9a-methoxymitosan, 580 mg (1.73 mmol) is placed in a round bottom flask and dissolved in 60 ml of methylene chloride. The triazing of Example 11, approximately 2.5 g (5.7 mmol) was added to the solution in the flask and the mixture was stirred at 5 ° C for 14 h and then at room temperature for 8 h. The progress of the reaction was monitored by thin layer chromatography on silicon oxide using methylene chloride methanol (9: 1). The reaction was allowed to stand at room temperature for an additional 26 hours and then worked up by column chromatography using a column packed with alumina which was 0.5 cm in diameter and 30 cm in length. The solvents used for the development and elution were 200 ml portions of methylene chloride, 0.5% methanol in methylene chloride, 1.0% methanol in methylene chloride, 1.5% methanol in methylene chloride, 2% methanol in methylene chloride and 4% Methanol in methylene chloride. The appropriate fractions were combined and evaporated to give the title compound, 470 mg.

Analysis for C22H23N5O8S2:

Calculated: C 45.65 H 4.09 N 11.82 (corrected for 0.5 mol CH2CI2)

Found: C 45.74 H 4.14 NI 1.61.

IR (KBr) vmax, cm-1: 3440-3200, 3060, 2930, 1720, 1570, 1510, 1395, 1335, 1210, 1055.

> H NMR (pyridine-ds, 5): 1.81 (s, 3H), 2.00 (bs, 1H), 2.61 (bs, IH), 2.98 (bs, IH), 3.08 (s, 3H), 3.20 (m, 2H), 3.39 (d, 1H), 3.83 (dd, IH), 4.07 (d, 1H), 4.59-4.89 ( m, 3H), 5.21 (dd, 1H), 7.16 (dd, IH), 8.31 (dd, IH), 8.71 (dd, 1H).

Example 2

The mitosane derivatives of the following formula can be prepared by adapting the processes of triazene preparation 1 and process 6 to other ca- (3-nitro-2-pyridyldithio) alkylamines having 2-6 C atoms in the alkyl group:

. n rf \ vss- (ch_) -0 1 ■ i n

ch.

n = 2-6

R '= H, or C' ~ 6 alkyl

Examples 3-16

The 7-alkoxydithio-9a-methoxymitosans 21-34 of Table V below were prepared according to Method A or B, described below, and reported in Table V. The physical data for Mit-osanverbindungen 21-34 are given in Table VI below.

Procedure A

To an oxygen-free solution of the mitosane of Example 20 (0.1 mmol) in acetone (3-5 ml) was added triethylamine (1.1 equivalents) with stirring and under argon 5 and then dropwise with a mercaptana (1 equivalent) in acetone (1-2 ml) was added. Most reactions b, the progress of the reaction is followed by thin layer chromatography on silica gel (10% CH3OH in CH2CI2). The completion of the reaction is indicated by the disappearance of the stain corresponding to the starting material and the appearance of the product stain. At this point the reaction mixture is concentrated under reduced pressure (at 30 ° C) and the residue is chromatographed on a neutral Woelm aluminum 15 oxide column (6 x 250 mm), which was packed with a slurry of 2-5% CH3OH in CH2CI2. This process separates the desired mitosane product from the pyridylthione by-product which is retained in the column. The product eluted using 2-5% CH3OH in CH2CI2 is then carefully purified by flash silica gel chromatography using 5-7% CH3OH in CH2CI2 as the eluent. The main band corresponding to the product is isolated and the amorphous 7-alkoxydithio-9a-methoxymitosan is characterized.

a If the starting mercaptan becomes impure, more than

1 equivalent of thiol required.

30 b If the starting mitosan according to Example 20 and the product in thin layer chromatography have Rr values that are very close to one another, high-pressure liquid chromatography (HPLC) is used for the monitoring (nBondapak-Cis column).

35

Procedure B

Saturated aqueous NaHCO 3 solution b (6 drops) is added to a solution of the mitosane from Example 20 (0.1 mmol) in 2-5% acetone 3 in methanol (10 ml), then 40 with mercaptan (1 equivalent) in methanol 0 (1 ml) was added. The progress of the reaction is followed by TLC (silica gel, 10% CH3OH in CH2CI2). After the reaction is complete, the reaction mixture is diluted with water (15 ml) and concentrated to 10 ml at 30 ° C under reduced pressure. 45 The resulting solution is chromatographed on a reverse phase C-18 column using a graded gradient elution (100% d H2O to 80% CH3OH in H2O). The product, which is eluted as the main red band, is collected and concentrated to give the 7-alkoxydithio-9a-5o methoxymitosan as an amorphous solid. If further purification is required, the above chromatography step is repeated.

a Methylene chloride can also be used, but 55 acetone is preferred.

b If the mercaptan is L-cysteine, this base is not used.

0 Water is used if the starting thiol is water-soluble.

60 d Elution with water separates the yellow pyridylthione by-product from the product, which is retained in the column.

17 667 094

Table V

7-AUtoxydithio-9a-methoxymitosane ch2ocnh2

A or HSS ^ ° Y RSH + 20 ->

g ru n i> 'H

Example thiol

No. (FSH) procedure product

1

3 ethyl 2-mercapto-A R = -CH2 CH2 OCCH ^ acetate

4 •. 3-mercapto-1,2-A R = -CH2CH (OH) CH2OH. propanediol

5 3-mercaptopropionic acid B R = -CH2CH2COO Na +

Cysteine B R = -CH2CH (NH3 +) COO "

7. Thiophenol

8 p-nitrobenzenethiol A R = -V> NO

■ O

2nd

p-methoxybenzenethiol A 0CH3

O-

10 p-aminobenzenethiol

A .R * \ N> -NH2 coo ~ Na +

11 2- Mercaptobenzoic acid B "R ■

12 2-Nitro-4-mercapto- B R »

benzoic acid

13 4-mercaptopyridine A R

667 094

18th

Example thiol

No. (ESH) process product

14 • 2-mercaptomethyl-1- • methylimidazole

R = -CH.

'- <0

N I

CH.

15

Glutathione

NH_

I - +

NHCOCH2CH2CHCOO Na B R = -CH, CH

2I - +

C0NHCH2C00 Na

16

Dime thy lami n-ethanethiol

25th

Table VI

R = -CH2CH2N (CH3) 2

7-alkoxydithio-9a-methoxymitosane

Example No.

formula

NMR data (pyridine -dg) a (5, ppm)

uv (ch3oh)

(X,

Max'

nm)

IR (KBr) <% ax '

R — CH2CH2

i

OCCH,

? "

R »-CH2CHCH2OH

520.324, 216

1.76 (s, 3H), 1.84 (s, 3H), 1.97 (t, lH, 7.9Hz), 2.59 (ba.lH), 2.87 (t, 2H, 6.7Hz), 2.96 (t, 2H, 6.2Hz) , 2.99 (bs, 1U), 3.07 (s, 3H), 3.37 (d, lH, 12.1Hz), 3.84 (dd, 1H, 11.2, 4.3Hz), 4.05 (d, lH, 12.1Hz), 4.27 (t , 2H, 6.7Hz), 4.56 (m, 2H), 4.92 (t, 1H, 10.8Hz), 5.25 (dd, 1H, 11.2,4.3Hz).

1.77 (s, 3H), 1.99 (bs, 1H), 2.60 (bs, lH), 215.324,

3.03 (t, 2H, 6.3Hz), 3.00 (m, lH), 3.07 (s, 3H), 3.15 (m, lH), 3.28 (dd, 1H, 4.5,13.2Hz), 3.37 (d, lH, 12.4 Hz), 3.84 (dd, 1H, 4.5.11.2Hz), 3.94 (bs, 2H), 4.06 (d, lH, 12.4Hz), 4.28 (bs, lH), 4.57 (m, 1H), 4.64 (m, 1H), 4.92 (lH, t, 10.6Hz), 5.25 (dd, lH), 6.29 (bs, lH), 6.69 (bs, 1H), 7.5 (m, 2H).

520

3370,3310,2930, 1740,1720,1640, 1560,1520,1335, 1220,1200,1065, 1030.

3440, 2965, 1720, 1655, 1635, 1580, 1455, 1410, 1340, 1305, 1215, 1070, 1020.

19th

667 094

Example XH NMR Data (Pyridine "d5) a UV (CHjOH) IR (KBr)

Formula (? »PPm> <Amax 'nm) {Vmax'

R — CH2CH2CCX> "Na +

10th

f2

r «ch2chcooh

R- ^

- <^) - N02

OCH,

NU-

11 R-

coo na

1.77 (s, 3H), 2.60 (bs, 1H), 2.78 (t, 2H, 215.323, 7.4Hz), 2.94 '(m, 3H), 3.00 {s, 3H), 510

3.17 (t, 2H, 7.4Hz), 3.38 (d, lH, 12.4Hz), 3.83 (dd, lH, 4.0.11.0Hz), 4.06 (d, lH, 12.5Hz), 4.58 (m, 2H), 4.84 (m, lH),

5. 20 (dd, 1H, 4.1, 10.6Hz), 7.64 (m, 2H).

1. 92 (s, 3H), 2.95-3.15 (m, 5H), 3.29 216.324, (s, 3H), 3.36 (dd, lH, 4.0.15.0Hz), 4.02 381.526 (d, lH, 13Hz), 4.10 (dd, lH, 4.0.8.7Hz), 4.30 (t, lH, 10.5Hz), 4.48 (m, 2H), 4.66 (dd, lH, 4.8.10.8Hz).

1.75 (s, 3H), 1.99 (bs, 1H), 2.61 (bs, lH), 206.324, 2.98 (m, 3H), 3.08 (s, 3H), 3.39 (d, 1H), 524 3.82 (dd, lH , 4.3.11.0Hz), 4.06 (d, lH, 12.4Hz), 4.49 (m, 1H), 4.57 (m, lH), 4.92 (m, lH), 5.23 (dd, lH, 4.3.11.0Hz ), 7.11 (d, 1H, 7.2Hz), 7.19 (dd, 2H, 7.2.7.2Hz), 7.50 (d, 2H, 7.2Hz).

I.73 (s, 3H), 1.94 (m, lH), 2.57 (bs, lH), 322.215 2.95 (bs, lH), 3.00 (t, 2H), 3.04 (s, 3H),

3. 35 (d, 1H, 12 '. 1Hz), 3.79 (dd, 1H, 4.2,

II.0Hz), 4.02 (d, lH, 12.1Hz), 4.53 (m, 2H), 4.86 (t, lH, 11.0Hz), 5.20 (dd, 1H, 4.2.11.0Hz), 7.55 (d, 2H, 8.8Hz), 8.02 (d, 2H, 8.8Hz).

1.73 (s, 3H), 2.00 (m, lH), 2.69 (bs, lH), 321.236

3.00 (bs, lH), 3.02 (t, 2H), 3.08 (s, 3H),

3.37 (d, lH, 12.0Hz), 3.51 (s, 3H),

3.84 (dd, 1H, 4.2.11.0Hz), 4.06 (d, 1H,

12.5Hz), 4.59 (m, 2H), 4.93 (t, lH,

10.6Hz), 5.27 (dd, 1H, 4.2.11.0Hz),

6. 84 (d, 2H, 8.7Hz), 7.50 (d, 2H, 8.7Hz).

I.74 (s, 3H), 2.05 (t, lH, 6.6Hz), 318.252 2.60 (bs, 1H), 2.96-3.11 (m, 3H),

3.07 (s, 3H), 3.37 (d, lH, 12.5Hz), 3.84 (dd, 1H, 4.2.11.0Hz), 4.09 (d, lH, Ì2.5Hz), 4.53 (m, 2H), 4.96 (t , lH, 10.6Hz), 5.41 (dd, lH, 4.2,

II.0Hz), 5.84 (bs, 1H), 6.72 (d, 2H, 8.2Hz), 7.37 (d, 2H, 8.2Hz):

1.76 (s, 3H), 1.95 («, 1H), 2.59 (bs, lH), 214.246,

2.90 (t, 2H, 6.2Hz), 2.95 (bs, 1H), 320.524

3.06 (s, 3H), 3.38 (d, lH, 12.8Hz),

3.80 (dd, lH, 4.2.11.0Hz), 4.05 (d, lH,

12.4Hz), 4.55 (m, 2H), 4.87 (m, lH),

5.18 (dd, 1H, 4.2,10.4Hz), 7.08 (t, lH,

7.4Hz), 7.23 (dd, lH, 7.8.7.4Hz),

7.5 (bs, 2H), 8.2S (d, 1H, 8.1Hz),

8.47 (d, 1H, 7.7Hz).

3430, 2920, 1715, 1650, 1620, 1575, 1450, 1405, 1340, 130.0, 1210, 1110, 1065.

3430, 2920, 1715, 1635, 1575, 1495, 1450, 1340, 1300, 1210, 1060.

3440.29 20.1715, 1650.1630.1575, 1450.1335.1300, 1210.1105.1055.

3460.3290.2940, 1715.1645.1620, 1570.1505.1445, 1400.1335.1295, 1210.1105.1055,

3460,3300,2930, 1715,1625,1570, 1490,1440,1400, 1330,1295,1245, 1210,1170,1100, 1055,1020.

3450.3370.29 20, 1720.1630.1575, 1495.14 50.1410, 1335.1300.1210, 1105.1065.

3390,3300,2930, 1720,1665,1580, 1555,1455,13 90, 1340,1305,1215, 1110,1070,10 25.

667 094

20th

Example No.

formula

XH NMR data (pyridine -dg) a (5, ppm)

UV (CH3OH)

Max'

nm)

IR (KBr) (vmax 'ö «" 1 »

12 R- - ^ - NO2

COO Well

13

14

-O

R = -CH

rCj)

N-I

CU,

1.75 (s, 3H), 2.58 (bs, IH), 2.93 (t, 2H, 215.321, 6.3Hz), 2.90 (m, 1H), 3.38 (d, 1H, 520 520 12.8Hz), 3.82 (dd , lH, 4.3,10.9Hz), 4.06 (d, lH, 12.1Hz), 4.53 (m, 2H), 4.88 (t, lH, 10.5Hz), 5.21 (dd, lH, 4.2, 10.7Hz), 7.45 ( d, 1H, 8.5Hz), 7.65 (d, lH, 8.3Hz), 8.14 (bs, lH).

1.76 (3H, s), 1.95 (bs, 1H), 2.60 (bs, 215.241,

1H), 3.01 (t, 2H, 6.1Hz), 3.00 (m, lH), 323.522

3.08 (9.3H), 3.39 (d, lH, 11.5Hz),

3.83 (dd, 1H, 4.2.11.0Hz), 4.07 (d, lH,

12.4Hz), 4. 52 (m, 2H), 4.90 (bs, lH),

5.25 (dd, lH, 4.3, 10.5Hz), 7.39 (dd,

2H, 1.5,4.5Hz), 7.6 (m, 2H),

8.48 (dd, 2H, 1.5.4.5Hz).

1.74 (s, 3H), 2.59 (bs, 1H), 2.73 (t, 2H, 218.323,

6.4Hz), 2.98 (d, lH, 4.2Hz), 3.08 (s, 5 20

3H), 3.40 (m, 1H), 3.43 (s, 3H),

3.84 (dd, 1H, 4.4.11.0Hz), 4.06 (d,

1H, 12.5Hz), 4.11 (a, 2H), 4.46 (m, 2H),

4.89 (m, lH), 5.25 (dd, 1H, 4.4,10.5Hz),

6.95 (q, 2H).

3440, 2930, 1720, 1620, 1580, 1520, 1450, 1400, 1340, 1215, 1070

3430, 2920, 1720, 1650, 1630, 1575, 1450, 1405, 1335, 1300, 1210, 1065.

3430, 2960, 1720, 1650, 1630, 1580, 1490, 1450, 1335, 1300, 1210, 1105, 1065.

15

16

r * _ +

JIHCOCH2CH2CHCOO Na R = »- CH, CH

I - +

CONHCHjCOO Well

R = —CHjCHjN (CH3) 2

1.91 (s, 3H), 2.16 (dd, 2H, 7.4,14.2Hz), 2.55 (dt, 2H, 2.3,8.2Hz), 2.95-3.10 (m, 5H), 3.28 (s, 3H), 3.28 (dd , lH, 4.6,14.1Hz), 3.61 (d, lH, 13.1Hz), 3.70 (dd, lH, 4.4,10.5Hz), 3.70-3.90 (m, 4K), 4.02 (d, 2H, 13.1Hz), 4.30 (t, lH, 10.5Hz), 4.46 (m, 2K), 4.65 (dd, 1H, 4.5,10.7Hz).

1.77 (s, 3H), 1.93 (bs, lH), 2.14 (3, 6H), 2.61 (m, 3H), 2.80-3.05 (m,. 5H), 3.08 (s, 3H), 3.40 (m, lH ), 3.84 (dd, 1H, 4.3.10.3Hz), 4.06 (d, lH, 12.5Hz), 4.57 (m, 2H), 5.26 (dd, lH, 4.3.10.3Hz)

326,366,524

3380.3290.2920, 1715.1650.1635, 1580.1455.1405, 1340.1300.1210, 1105.1065.1030.

a) D20 was used for the mitosans according to Examples 6 and 15.

G

Claims (24)

667 094 2nd PATENT CLAIMS 1. Compounds of formula II or III ii t-r ' ch r-ss-alk., - iii f-r ' ce. bliss Alki is straight or branched chain alkylene, with 1-6 C atoms if R3 is attached to it via a carbon atom, or with 2-6 C atoms if R3 is attached to it via sulfur, oxygen or nitrogen, and R3 and -SS- in this case are attached to different C atoms , Alk2 is straight or branched alkylene, with 2-6 C atoms, which optionally bears a substituent A, to which the sulfur and oxygen atoms are bonded and any A substituent present thereon to various C atoms via oxygen, sulfur or nitrogen Alkì is bonded, the said substituent A being selected from 1 or 2 members of the group C ^ -alkyl, C] _6-alkanoyl, Ci_6-alkoxy, halogen, C, "6-alkoxycarbonyl, cyano, Cj ^ -alkylamino, Ci_6 -Dialkylamino, C | _6-alkanoyl-amino and C i _6-AI koxycarbonyl, Alki and Alk2 may contain a double bond, R1 denotes hydrogen, lower alkyl, lower alkanoyl, benzoyl or substituted benzoyl, where the substituent is selected from lower alkyl, lower alkoxy, halogen, amino or nitro, R3 is selected from the group consisting of halogen, carboxy, alkanoyloxy with 1-7 C atoms, hydroxy, the oxygen atom being bonded to alki with 3-6 C atoms, alkylamino or dialkylamino with 1-12 C atoms, N. -Al-koxy-alkylamino with 2-7 C atoms, alkanoylamino with 1-7 C atoms, benzoylamino or B-substituted benzoyl-amino, naphthoylamino or B-substituted naphthoyl-amino, phenylamino or B-substituted phenylamino, cycloalkyl or B-substituted cycloalkyl with 3-8 ring members, cycloalkenyl or B-substituted cycloalkenyl with 5-8 ring members, phenyl or B-substituted phenyl, naphthyl or B-substituted naphthyl, heterocyclic group selected from the group heteroaromatic and heterocyclic groups with 1- 2 rings, with 2-8 ring members in each ring and 1-2 heteroatoms in each ring, selected from oxygen, nitrogen and sulfur, pyridylamino or thiazolylamino, alkoxy or alkylthio with 1-6 C atoms, alkoxycarbonyl or alkylaminocarbonyl with 2-7 atoms each, aminocarbonyl, phenoxycarbonyl or B-substituted phenoxy-carbonyl, phenoxy or B-substituted phenoxy, naphthoxy or B-substituted naphthoxy, alkoxycarbonylamino with 2-6 C atoms, ureido of the formula -NHCONH2, N -Alkyl- ureylene of the formula -NHCONHalkyl with 2-7 C atoms, N3-haloalkylureylene with 3-7 C atoms, N3-haloalkyl-N3-nitrosoureylene with 3-7 C atoms, dialkylaminocarbonyl with 3-13 C atoms, dialkylaminoalkoxy with 4-13 C-Ato-men, alkanoylaminoalkoxy with 3-7 C-atoms and hydroxy-alkylamino or N, N-dihydroxyalkylamino with 2-8 C-atoms each, wherein said substituent B 1 or 2 members of the group lower alkyl, lower alkanoyl , Lower alkoxy, halogen, amino, carboxy, hydroxy and nitro, and R4 is selected from the group alkyl with 1-12 C atoms, alkenyl or alkynyl with 3-12 C atoms, cycloalkyl or B-substituted cycloalkyl with 3-8 ring members, cycloalkenyl or B-substituted cycloalkenyl, each with 5-8 Ring members, phenyl or B-substituted phenyl, naphthyl or B-substituted naphthyl, heterocyclic group selected from heteroaromatic and heteroalicyclic groups with 1-2 rings, each with 3-8 ring members per ring and 1-2 heteroatoms per ring, selected from oxygen, nitrogen and sulfur, with the proviso that the heterocyclic group is connected by a carbon atom which is bonded to at least one other carbon atom, the substituent B being one or two radicals from the group lower alkyl , Lower alkanoyl, lower alkoxy, halogen, amino, carboxy, hydroxy or nitro and R4 and the adjacent S atom together can mean S-cysteinyl, where the said S-cysteinyl group can be esterified, salified or linked to a non-toxic and non-allergic peptide or a non-toxic, pharmaceutically acceptable salt thereof. 2. A compound according to claim 1, wherein Alla is ethylene and R1 is hydrogen. 3. A compound according to claim 2 of formula II, wherein Alki is ethylene and R3 is acetylamino. 4. A compound according to claim 2 of formula II, wherein Alki is ethylene and R3 is acetyloxy. 5. A compound according to claim 2 of formula II, wherein Alki is methylene and R3 is 1,2-dihydroxyethyl. 6. A compound according to claim 2 of formula II, wherein Alki is ethylene and R3 is carboxy or a non-toxic, pharmaceutically acceptable salt thereof. 7. A compound according to claim 2 of formula II, wherein Alki is methylene and R31-carboxyaminomethyl or a non-toxic pharmaceutically acceptable salt thereof. 8. A compound according to claim 2 of Formèl II, wherein Alki is methylene and R3 is l-methylimidazol-2-yl. 9. A compound according to claim 2 of formula II, wherein Alki is ethylene and R3 is dimethylamino. 10. A compound according to claim 2 of formula III, wherein R4 is phenyl. 11. A compound according to claim 2 of formula III, wherein R4 is 4-nitrophenyl. 12. A compound according to claim 2 of formula III, wherein R4 is 4-methoxyphenyl. 13. A compound according to claim 2 of formula III, wherein R4 is 4-aminophenyl. 14. A compound according to claim 2 of formula III, wherein R4 Is 2-carboxyphenyl or a non-toxic, pharmaceutically acceptable salt thereof. 15. A compound according to claim 2 of formula III, wherein R4 is 4-nitro-3-carboxyphenyl or a non-toxic, pharmaceutically acceptable salt thereof. 16. A compound according to claim 2 of formula III, wherein R4 is 4-pyridyl. 17. A compound according to claim 2 of the formula 5 10th 15 20th 25th 30th 35 40 45 50 55 60 65 667 094 ? H2 H02CCHCH2CH2C0NH H02CCH2NHC0 or a non-toxic, pharmaceutically acceptable salt CHCH _SSCHnCH-, 0 I 2 24. A process for the preparation of a compound according to wherein R1 and Alla are as defined in claim 1. 19. A compound according to claim 2 of formula XVII SS-CH2CH2-0, 25th 30th 35 XVII 20. A compound according to claim 1 as an agent for inhibiting a tumor in humans or mammals. 21. A pharmaceutical composition containing a compound according to claim 1 together with a pharmaceutically acceptable solvent, diluent, additive or carrier. 22. A process for the preparation of a compound according to claim 1, characterized in that at least one equivalent of a triazene of the formula V or VI Ar-N = N-NH-Alk2-SS-Alki-R3 Ar-N = N-NH-Alkî-SS-R4 with one equivalent of Mitosan of formula IV O II CH-OCNH- (V) (VI) 45 50 IV wherein R1, R3, R4, Alki and Alkî have the meaning given in claim 1, is reacted in an inert solvent and at a temperature of 0-60 ° C. 25. The method of claim 24, wherein Alla is ethylene and R1 is hydrogen. 26. The method of claim 24 or 25, wherein one equivalent of a base is used in the reaction. 27. The method according to claim 24 or 25, wherein a lower alkanol, a lower alkyl ester of a lower alkanoic acid, a lower aliphatic ketone, a cyclic aliphatic ether or a lower polyhalogenated, aliphatic hydrocarbon with up to 8 C atoms or water is used as the reaction medium.