SE465929B - SUBSTITUTED 7-OXOMITOSANES - Google Patents

Description

465 929 10 15 20 25 30 35 A trivial nomenclature system, which has found widespread use in the mitomycin literature, identifies the foregoing ring system comprising several of the characteristic substituents in the mitomycins such as mitosan.

Mitosan According to this system, mitomycin A is 7,9a-dimethoxymitosane and mitomycin C is 7-amino-9a-methoximitosan. Regarding the stereochemical configuration of the products of the present invention, these are identified by the parent expression "mitosan" or by structural formulas for identifying the stereochemical configuration thereof in the same manner as for mitomycin A or C. Û CH20CONH2 Mitomycin A g = R1 = 0CH Mitomycin c R; 2 I R1-: AND 3 Ä / 10 15 20 25 30 35 465 929 'Mitomycin C is an antibiotic, which is produced by fermentation and is currently commercially available with permission from the Food and Drug Administration for therapy of disseminated adenocarcinoma of the stomach or pancreas in tried combinations with other approved chemotherapeutic agents and as palliative treatment when other therapies have failed (Mutamyciášïßristol Laboratories, Syracuse, New York 13221, Physicians' Desk Reference 37th Edition, 1983, pp. 747 and Mitomycin C and its preparation by fermentation are the subject of U.S. Patent 3,660,578, issued May 2, 1972.

The structures of mitomycin A, B, C and of porfiromycin were first published by J. S. Webb et al. at Lederle Laboratories Division American Cyanamid Company, J. Am.

Chem. Soc., 84, 3185-3187 (1962). One of the chemical transformations used in this structural study to relate mitomycin A to mitomycin C was the conversion of the former, 7,9a-dimethoxymitosan, by reaction with ammonia to the latter, 7-amino. - 9% -methoxymitosan. The repression of the 7-methoxy group in mitomycin A has been found to be a reaction of considerable interest in the preparation of active antitumor derivatives of mitomycin C. Recently, the stereochemical configurations of positions 1, 1a, 8a and 8b have been found to be those set forth above with regarding the nomenclature according to Chemical Abstracts [Éhirhata et al., J. Am. Chem. Soc., 105, 7199-7200 (1983)]. The previous literature refers to the enantiomer.

The following articles and patents refer to e.g. the conversion of mitomycin A to a 7-substituted derivative of aminomitomycin C with antitumor activity. The aim of this study was to produce derivatives which were more active and in particular which were less toxic than mito- «: mycine C: 465 929 10 15 20 25 30 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); 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. Pat. 3,332,944 of July 25, 1967; Matsui et al., U.S. Pat. 3,420,846 of January 7, 1969; Matsui et al., U.S. Pat. 3,540,705 of June 17, 1969; Matsui et al., U.S. Pat. 3,514,452 of May 26, 1970; Nakano et al., U.S. Pat. 4,231,936 of November 4, 1980.

Remers, U.S. 4,268,676 of May 19, 1981.

The following patent applications relate to the preparation of 7-substituted derivatives of aminomitomycin C, wherein the substituent includes a disulfide bond.

Kono et al., European Patent Application No. 116 208 (1984); Vyas et al., British Patent Application No. 2,140,799 (1984). 7-alkoxy-substituted mitosanes, which are structurally related to mitomycin A, are described as useful antibiotics with activity on tumors in experimental animals according to an article by Urakawa et al., J. Antibiotics, 23, 804-809 (1980).

Mitomycin C is the main mitomycin product produced by fermentation and as the most widely available form. Current technology for the conversion of mitomycin C to mitomycin A has a number of shortcomings. Hydrolysis of mitomycin C to the corresponding 7-hydroxy-9a-methoxymitosan and then methylation of this substance requires diazomethane, a very poor substance to handle on a production scale, and the 7-hydroxy intermediate is very unstable [Matsui et al., J. Antibiotics, XXI, 189-198 (1968)]. An attempt to avoid these difficulties involves the use of 7-acyloximitosants (Kyowa Hakko 10 15 20 25 30 35 465 929 'Kogyo KK Japanese Patent J5 6073-085, Farmdoc No. 56227 D / 31). Alcoholysis of mitomycin A, as described by Urakawa et al., J. Antibiotics, 23, 804-809 (1980), is limited to the preparation of only specific 7-alkoxy structure types due to the availability and reactivity of the alcohol starting materials.

The present invention relates to a group of mitomycin A analogs having an organic dithio substituent incorporated with the alkoxy group in the 7-position. These compounds can be illustrated by the general formula R 2 ° cnzocrmz a -ss-Aikz-o wherein R 2 is an organic group, namely the structural component of an organic thiol of the formula R 2 SH, and Alk 2 and R 1 have the meanings given below. These compounds may alternatively be described by formulas II and III. wherein Alk1 Alk Alk R3 is halogen, carboxy, alkanoyloxy having 1-7 carbon atoms III is a straight or branched chain alkylene group having 1-6 carbon atoms when R3 is joined thereto via a carbon atom thereof and having 2-6 carbon atoms when R 3 is joined thereto via a sulfur, oxygen or nitrogen atom thereof and R 3 and -SS- in this case are attached to different carbon atoms; is a straight or branched chain alkylene group having 2 to 6 carbon atoms, optionally bearing a substituent A, the sulfur and oxygen atoms attached thereto and an optional substituent A, which is attached thereto via oxygen, sulfur or nitrogen, being linked to different carbon atoms in Alk 2, wherein the substituent A is one or two C 1-6 alkyl, C 1-6 alkanoyl, C 1-6 alkoxy, halogen, C 1-6 alkoxycarbonyl, cyano, C 1-6 alkylamino, C 1-6 dialkylamino, C 1-6 alkanoylamino or C 1-6 alkanoylamino; and Alkz may contain a double bond; is hydrogen, lower alkyl, lower alkanoyl, benzoyl or substituted benzoyl, wherein the substituent is lower alkyl, lower alkoxy, halogen, amino or nitro; more, hydroxy wherein the oxygen atom is bonded to Alk1 alkylamino having 1-12 carbon atoms, N-alkoxyalkylamino having 2-7 carbon atoms, alkanoylamino having 1-7 having 3-6 carbon atoms, alkylamino or dicolatoms, benzoylamino or B-substituted benzoylamino , naphthoylamino or B-substituted naphthoylamino, phenylamino or B-substituted h, phenylamino, cycloalkyl or B-substituted cycloalkyl each having 3-8 ring members, cycloalkenyl or B-substituted cycloalkenyl each having 5 -8 ring, phenyl or B-substituted phenyl, naphthyl or B-substituted naphthyl, a heteroaromatic or heteroalicyclic group having 1-2 rings, 3-8 rings in each ring and 1-2 heteroatoms in each ring, which heteroatom consists of oxygen, nitrogen and / or sulfur, pyridylamino or thiazolylamino, alkoxy or alkylthio each having 1-6 carbon atoms, alkoxycarbonyl or alkylaminocarbonyl each having 2-7 carbon atoms, aminocarbonyl, phenoxycarbonyl or B-substituted phenoxycarbonyl, phenoxy B-su substituted phenoxy, naphthoxy or B-substituted naphthoxy, alkoxycarbonylamino having 2-6 carbon atoms, ureido (-NHCONH2), N-alkylureylene (-Nnconnealkyl) having 2-7 carbon atoms, N3-whole-alkylureylene having 3-7 carbon atoms, N3 -haloalkyl- N3-nitrosoureylene having 3-7 carbon atoms, dialkylaminocarbonyl having 3-13 carbon atoms, dialkylaminoalkoxy having 4-13 carbon atoms, alkanoylaminoalkoxy having 3-7 carbon atoms and hydroxyalkylamino or N, N-dihydroxyalkylamino each having 2-8 carbon atoms, the B substituent being one or two lower alkyl, lower alkanoyl, lower alkoxy, halogen, amino, carboxy, hydroxy or nitro groups, and R 4 is alkyl of 1-12 carbon atoms, alkenyl or eikynyl each with 3-12 carbon atoms, cycloalkyl or B-substituted cycloalkyl having 3-8 ring links, cycloalkenyl or B-substituted cycloalkenyl each having 5-8 ring links, phenyl or B-substituted phenyl, naphthyl or B-substituted naphthyl, a heteroaromatic or heteroalicyclic group having 1-2 rings, 3-8 ring members in each ring and 1-2 hot 465,929 heteroatoms in each ring, which are 465,929 atoms are oxygen, nitrogen and / or sulfur, with the proviso that the heterocyclic group is linked via a carbon atom, which is bonded to at least one other carbon atom (ie the carbon atom). atom attached to -SS- may not itself be attached to two other heteroatoms), the B substituent being one or two lower alkyl, lower alkanoyl, lower alkoxy, halogen, amino, carboxy , hydroxy or nitro groups, or R 4 and the adjacent sulfur atom together form S-cysteinyl, wherein the S-cysteinyl group may be esterified, in salt form or in a non-toxic and non-allergenic peptide, and non-toxic, pharmaceutically acceptable salts thereof.

The compounds of the present invention are inhibitors of experimental tumors in animals. The substances which in the present context are referred to as the compounds according to Examples 17, 20 and 21-34 are more specifically new substances. They were used in a manner similar to the use of mitomycin C. The dosages used are adjusted according to their toxicity relative to the toxicity of mitomycin C1. In cases where the new compound is less toxic, a higher dose was used.

According to a further aspect of the present invention there is provided a novel process for the preparation of mitosanes of formulas II and III. This new process involves reacting a mitosane of formula IV O 0 CN CN Hzm fl z 35 H and IV cH -R 1 'n with a triazene of formula V or formula VI Ar-N = N- Nu-A1k2-ss-A1k1-R3 Ar-N = NH-A1k2-ss-R4 V VI. 1 3 4. wherein R, R, R, Alk1 and Alkz have the meanings given above and Ar is the organic residue of a diazotizable aromatic amine.

According to a variant of the present invention, there is provided an alternative process for the preparation of mitosanes of formulas II and III. This process involves reacting a thiol of formula VII or VIII R3Alk1SH R4SH VII VIII with a mitosan derivative of formula Ib NO 2 0 oi / \ss-Alkz-o C32 NH? AND sr 3 Ib C113 _R The disulfide mitosanes of formula Ib are prepared by the triazene method described below. The mitosans of formula Ib, wherein Aikz is erylene and R1 is hydrogen, are described in more detail in Example 20.

According to another aspect of the present invention there is provided an improved process for the preparation of compounds of formula IX wherein R 5 is hydrogen or C 1-6 alkyl and R 6 is C 1-12 alkyl or substituted C 1-12 alkyl, C 3-12 -cycloalkyl or substituted C 3-12 cycloalkyl, wherein the carbon atom attached to the mitosane-7-oxygen atom has 1-2 hydrogen atoms and the substituents are halogen, C 1-6 alkoxy, C 1-6 alkanoyl, C 1-4 alkylamino, C 2-12- dialkylamino, C 6-12 aryl, C 6-12 aroyl, cyano, trihalomethyl, amino, C 1-6 monoaryloxy, C 1-6 alkanoyloxy, C 7-14 aroyloxy, 1 or 2 ring heterocycle and 5-12 ring atoms containing up to 4 heteroatoms , which consists of nitrogen, oxygen and / or sulfur, and wherein the alkoxy, alkanoyl, aroyl, aryl, aryloxy, alkanoyloxy, aroyloxy and heterocycle substituents optionally contain 1-2 substituents, which consist of halogen, C1-6 alkoxy, C 1-6 alkanoyl, cyano, trihalomethyl, amino, C 1-6 alkylamino or C2-12 dialkylamino.

Many of the compounds of formula IX are known compounds with inhibitory activity with respect to experimental animal tumors in vivo. A number of novel compounds of formula IX have also been prepared by this process and fall within the scope of the present invention. In particular, the substances hereinafter referred to as the compounds of Examples 14, '15, 16 and 19 are novel substances and these also exhibit antitumor activity with respect to experimental animal tumors. These compounds fall within the scope of the present invention. They are used in a manner similar to the use of mitomycin C. The dosages used are adjusted relative to their toxicities relative to the toxicity of mitomycin C. In cases where the compound is less toxic, a higher dose.

The new process for the preparation of compounds of formula IX involves reacting a mitosane of formula X CH with a triazene of formula XI 6 Ar-N = N-NH-R XI wherein R 5 and R 6 have the meanings given above and Ar is the organic residue of a diazotizable aromatic amine.

The terms "lower alkyl", "lower alkoxy" and "lower alkanoyl" as used herein (unless otherwise indicated) mean straight or branched chain alkyl, alkoxy or alkanoyl groups having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl , butyl, isobutyl, t-butyl, amyl, hexyl, etc. Preferably these groups contain 1-4 carbon atoms and in particular they contain 1 or 2 carbon atoms, unless otherwise stated in any particular case the term "halogen" is intended to include chlorine, fluorine, bromine and iodine The term "non-toxic pharmaceutically acceptable salt" is intended to include 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, sulfonic acid, phosphoric acid, nitric acid, maleic acid, fumaric acid, succinic acid, oxalic acid, benzoic acid, methanesulfonic acid, tartaric acid, citric sulfonic acid and similar. Such bases are well known and include, for example, non-toxic metal salts such as sodium, potassium, calcium and magnesium, the ammonium salt and salts with non-fatty amines, for example trialkylamines, procaine, dibenzylamine, pyridine, N-methylmorpholine, N-methylpiperidine and the like. The salts are prepared by methods known in the art.

As stated above, the present invention provides a novel process for the preparation of compounds of formula IX, which process comprises reacting a mitosan of formula X with a triazene of formula XI as shown in Reaction Scheme 1.

Reaction Scheme 1 XI wherein R 5 and R 6 have the meanings given above and Ar is the organic residue of a diazotizable aromatic amine.

The 1-substituted-3-aryltriazenes of formula XI, and more particularly the 1-alkyl-3-aryltriazenes, constitute a class of reagents known to be useful for reaction with carboxylic acids to give corresponding lower alkyl esters. 1-Methyl-3- (4-methylphenyl) triazene can be prepared according to the general procedures described by E. H. White et al. In Org. Syn., 48, 102-105 (1968) and described in Preparative Example 1 below. This process can only be carried out with water-soluble amines and a second process, which is described by E. H. White et al., Tetrahedron Letters, No. 21, 761 (1961) and which is also described in Preparative Example 2 below, is more suitable for the preparation of triazenes of water-insoluble amines.

The reagent 1-methyl-3- (4-methylphenyl) triazene, prepared in the above manner, has been previously used for the preparation of methyl esters of carboxylic acids such as 2,4-dinitrobenzoic acid [E. H. White et al. Org. Syn., 48, 102-105 (1968)] and cephalosporanic acids which give the desired A3 compound without isomerization of the γ β-isomer [Mangia, Tetrahedron Letters, No. 52, pp. 5219-20 (1978)]. The reagent has also been used to prepare a 3-methoxy-cephalosporin derivative by reaction with the corresponding 3-hydroxy-3-cephem-4-carboxylate in benzene solution at reflux temperature (Wiederkeher et al. U.S. 4,069,324, issued January 17, 1978). ).

Other 1- (lower alkyl) -3-aryltriazenes of formula XI can likewise be prepared by reacting other lower alkylamines with aryldiazonium salts in a similar manner.

Each arylamine having 6-12 carbon atoms, which readily forms a diazonium salt, can be used as the source of the aryl moiety in the 1,3-disubstituted triazene. Some examples of triazenes prepared in this manner and used in accordance with the present invention are: 1- (n-butyl) -3- (4-methylphenyl) triazene; 1- (1-methylethyl) -3- (4-methylphenyl) triazene; 1- (4-methylphenyl) -3- [E- (4-morpholinyl) ethylphthriazen; 1- (4-methylphenyl) -3- [2- (2-pyridyl) ethyl] triazene; 1- (2-benzylthioethyl) -3- (4-methylphenyl) triazene; 1- (4-chlorophenyl) -3- (2-methoxyethyl) triazene; 1- (4-chlorophenyl) -3- (1,3-dioxol-2-ylmethyl) triazene; 1- (4-chlorophenyl) -3- (tetrahydrofuran-2-ylmethyl) triazene.

Other triazenes have been described in the literature, which are more suitable reactants for use in the process of the present invention to provide 7- (substituted alkoxymitosane) of formula IX. Those described by T. A. Daniels et al., Can. J. Chem., 55, 3751-3754 (1977) are examples thereof.

X - <::::> - N = N-NH-CH2-Y = H, Y = CN = NO2, Y = CN = C02Me, Y = CN = Ao, Y = CN - NO2, Y - C02Et = C02Me , Y = CO2Et = CO 2 Me, Y COPh = NO 2, Y Ul fl l-hß fl- OUW NNNNNNNN I ll .Il -CH (OCH3) 2 The following are further examples of suitable triazene starting materials of formula XI for use in the present invention. 1- (n-butyl) -3- (f-'-naphthyl) triazene 1- (n-hexyl) -3-phenyltriazen 1-ethyl-3- (2,4 = dimethylphenyl) triazene 1- (1-methylethyl ) -3- (4-methoxyphenyl) triazene For the preparation of mitomycin A, they prefer to use 3-methyl-1- (4-methylphenyl) triazene as methylation reagent. Preferably at least two molar portions of the latter are used per molar portion of the 7-hydroxy-9α-methoxymitosane and the reaction is preferably carried out in a liquid organic solvent for the 7-hydroxy-9α-methoxymitosane starting material. Preferred solvents are lower alkanols, lower alkanoic acid (lower alkyl alkoxy, di (lower alkyl) ethers, cyclic aliphatic ethers and lower polyhalogenated aliphatic hydrocarbons. These solvents contain up to 6 carbon atoms but those boiling at temperatures below 100 ° C are especially preferred. methylene chloride, methanol, diethyl ether, ethyl acetate and mixtures thereof The reaction can be carried out at the reaction mixture at reflux temperature or up to about 60 DEG C. At temperatures exceeding this, the mitosane reactant tends to decompose with an accompanying yield reduction. at room temperature or below, for example in the range 0-25 ° C.

A convenient way to determine whether the reaction is complete is by thin layer chromatography. Mitomycin A is dark purple and can be easily distinguished from the starting material method. In the solvent system methylene chloride / methanol (90/10), mitomycin A shows Rf = 0.36. Chromatography on neutral alumina can be used to purify the product.

The above reaction conditions and precautions are generally applicable to the preparation of other 7-R 60 mitosanes of formula IX according to the present process.

The novel process of the present invention utilizing 1-substituted-3-aryltriazenes can also be used to prepare compounds of formulas II or III, reacting a mitosane of formula IV with a triazene of formula V or VI as indicated in Reaction Scheme 2.

Reaction Scheme 2 Ar-N = N-NH-Alkz-ss-Alk1-R3 _, II V> II or 111 or Ar-N = N-NH-Alkz-SS-R4 VI wherein R1, R3, R4, Alk1 and Alkz has the meanings given above and Ar is the organic residue of a diazotizable aromatic amine.

Aryltriazenes of formulas V or VI can be prepared in a similar manner as described above for the preparation of aryltriazenes of formula XI except that the alkylamines used therein are replaced by amino disulfides of formula XII R2-SS-Alk NH XII 2 2 which alternatively can be described by formulas XIII and XIV 3 R -Alk1-SS-Alkz-NH2 And R -SS-Alkz-NH2 XIII XIV 10 15 20 25 30 35 17 465 929 'Aminodisulfides of formulas XIII and XIV are known compounds and can be prepared by various methods.

For example, they can be prepared by reacting the appropriate thiol R Alk1SH or R4SH with a Bunte salt of the formula NH2Alk2SSO3Na XV or with a sulphenylthiocarbonate of the formula OI NH2Alk2SSáOCH3 XVI Klayman et al., J. Org. Chem., 29, 3737-3738 (1964) have prepared the following compounds by the Bunte salt method: 2-aminoethyl-n-butyl disulfide; 2-aminoethyl-n-hexyl disulfide; 2-aminoethyl-n-octyl disulfide; 2-aminoethyl-n-decyl disulfide; 2-aminoethyl-phenyl-disulfide; 2-Aminoethyl-benzyl disulfide.

Methanol has been found to be the preferred reaction solvent for reacting the Bunte salt with the thiol.

Reaction temperatures of from 0 DEG to -10 DEG C. have been found to be preferred using this solvent. Higher temperatures were necessary with other solvents. The major disadvantage of this method is the formation of symmetrical disulfides as a by-product, probably as a result of disproportionation of the desired mixed disulfide.

The mixed disulfide starting materials of formulas XIII and XIV are preferably prepared via reaction of the appropriate thiol with a sulfenyl thiocarbonate of formula XVI. This is the method set forth by S. J.

Brois et al., J. Am. Chem. Soc., 92, 7629-7631 (1970).

Typically, this preparation process involves adding the thiol to a methanol solution of the aminoalkylsulphenylthiocarbonate of formula XVI and allowing the reaction to proceed at a temperature in the range of 0-25 ° C. The reaction times vary from mainly instantaneous turnover to several hours depending on the particular thiol in question used. The course of the reaction can be monitored by measuring the presence of unreacted thiol in the reaction vessel. If the reaction is slow, a catalytic amount of triethylamine can be added as a reaction accelerator. The 1- (substituted disulfide) -3-aryltriazines of formulas V or VI are prepared by reacting the amino disulfide of formula XII with aryldiazonium salts in a manner similar to that described above for the preparation of aryltriazenes of formula XI. Each arylamine 6-12 carbon atoms, which readily form a diazonium salt, can be used as the source of the aryl moiety in the 1,3-disubstituted triazine. Some examples of disulfide triazenes prepared in this manner and used in the present invention are 1- [2- (2-acetamidoethyldithio) ethyl] -3- (4-methylphenyl) triazene); 1- [2- (3-nitro-2-pyridyldithio) ethyl] -3- (4-methylphenyl) triazene.

Below are further examples of suitable triazene starting materials of formulas V or VI for use in the present invention. 1- [2- (3-nitro-2-pyridyldithio) ethyl] -3- (4-chlorophenyl) triazene; 1- [E- (3-nitro-2-pyridyldithio) propyl] -3- (4-methylphenyl) triazene; 1- [2- (z-pyridylethyl) ethyl] -5- (4-methylphenyl) triazene; 1- [2- (phenyldithio) ethyl] -3- (4-methylphenyl) triazene; 1- [1- (butyldithio) ethyl] -3- (4-methylphenyl) triazene; 1- [1- (4-methoxyphenyldithio) ethyl] -3- (4-methylphenyl) triazene; 1- [2- (4-nitrophenyldithio) ethyl] -3- (4-methylphenyl) triazene; 1- {E- [12-benzoylaminoethyl) dithio] ethyl} -3- (4-methylphenyl) triazene; 1- [2- (4-chloro-2-naphthyldithio) ethyl] -3- (4-methylphenyl) -triazene; 1% [N- (cyclopropylmethyldithio) ethyl] -3- (4-methylphenyl) triazene; 1- {2 - [(2-phenoxyethyl) dithio] ethyl} -3- (4-methylphenyl) triazene.

In a preferred embodiment of the present invention there is provided an alternative process for the preparation of disulfide mitosanes of formula Ia ° CHZOÉNHZ e ° CH3 2 R SS-CHZCHZO Ia CH3 N wherein R2 is an organic group, namely the structural component of an organic thiol of formula R 2 SH, which may alternatively be described by R 3 Alk 1 or R 4, where R 3, R 4 and Alk 1 have the meanings given above.

For the preparation of the disulfide mitosanes of formula Ia, it is preferred to use 9α-methoxy-7- [E- (3-nitro-2-pyridylthio) ethoxy] mitosane of formula XVII in a thiol exchange process with a suitable organic thiol of formula RZSH, as shown in Reaction Scheme 3. The driving force behind the formation of the disulfides of formula Ia is the stability of the by-product, namely 3-nitro-2-mercaptopyridine, which exists only as ion XVIII. 20 465 929 10 15 20 25 30 35 Reaction scheme 3 ° 2 o cazoäuaz y _- N \\ 3 C83 XVII 2 ° ca 03x32 - QR -SS-CH2CH2O * \\ 2 JIZH a 3 + N C33 H XVI Ia If it is It is desirable to prepare mitosanes of formula II or III, wherein Alkz has other meaning than ethylene, such as trimethylene or propylene, alternatively the appropriate triazene of formula V or VI is used in the process set forth in Reaction Scheme 2 to prepare disulfide mitosanes of formula Ib I m2 ° n oími / \ r 0 ° = = SS-A '2 §ocx3 Ib 31 i -n C33 10 15 20 25 30 35 21 465 929' wherein Alk2 and R1 have the meanings given above.

There are two general synthetic procedures, described in more detail below, for the preparation of both lipophilic and hydrophilic mitosanes of formula Ia. General method A is used for the preparation of either lipophilic or moderately soluble disulfides of formula Ia, while general method B is used for water-soluble disulfides of formula Ia, which are preferably isolated as the sodium salts or as the zwitterionic forms. Preferably, at least one equivalent of the mercaptan R2SH per equivalent of mitosan of formula XVII is used and the reaction may be carried out in the presence of about one equivalent of base per equivalent of mercaptan R2SH. Preferred bases are the tertiary amines, for example triethylamine, N-methylmorpholine, N-methylpiperidine, pyridine and 2,6-lutidine, and inorganic bases, for example sodium bicarbonate, potassium carbonate, potassium bicarbonate and the like. Suitable inert solvents for reacting the starting materials of formulas XVII and R 2 SH are lower alkanols, lower alkanoic acid (lower alkyl) esters, lower aliphatic ketones, cyclic aliphatic ethers, lower polyhalogenated aliphatic hydrocarbons and water. The organic solvents contain up to 8 carbon atoms, but those which boil at temperatures below 10,000 are preferred. Specifically 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 about 60 ° C. It is preferred to carry out the reaction at room temperature or below, for example in the range 0-25 C.

The above reaction conditions and precautions are generally applicable to the preparation of other disulfide mitosanes of formulas Ia and Ib according to the general procedure set forth in Reaction Scheme 3. 22 465 929 10 15 20 The following is a list of representative thiols of the formula R3Alk1SH or R4SH , which via the Bunte salt XV or the sulphenylthiocarbonate XVI can be converted into intermediates of formulas XIII and XIV, which in turn are converted into products according to the present invention. In the case of the preferred embodiment, the representative thiols can be used in a reaction with mitosanes of formulas Ia or Ib to produce products of the present invention.

However, the only limitations placed on the process of the present invention are the use of thiols which contain terminal primary alkylamines which may lead to a mixture of products, and the use of heteroaromatic thiols which may not react with compounds of formulas Ia or Ib . 23 qscn3 HSCHZCHB HSCH2CH2CH3 HSCH (CH3) 2 HS (CH2) 3CH3 HS ~ $ H-CH CH CH 2 3 3 nsca2cH (ca3) 2 HS “<3 465 929 ',' 465 9” l HS HS-CH HS-CH2 -CH = GH2 Hs-ca -cH = c (cH3) 2 2 HS-CH fC5CH 2 HS-CH -C5C-CH 2 3 .1 1 U Hs (cH2) noR n = 2-4; R = H, ccH3, cH3 Hs (cn2) ncxR n = 1-3; x = 0, NH, NR1; R / R1 ~ = -H, C33 W 1 _ _ _ 1 = Hs (cH2) nNHR n - 2 4, R CH3, cH2cH3, cnzcnzc fl s, c% g3 Hs (cH2) nNR1R2 n = 2-4; nl / R2 = CH3, bH2cH3 ns-cH2cH2scn3 Hs-cH2cH2NHc (CH3) 3 H Hs-CH -c-än NHR1 nl - ca 3 H3 Hs-cn cn NH 2 2 25 465 929 ca: as-cnzcnz-N \\ I n = 3-5 cnznnlaz as-Én-ca Nnlaz R1 = ca - az = H cn 2 3 '' 3 ns-c fl -cn-cs NHR1 nl - cn 2 2 '3 1 1 NHR HS-CI-Iz- çl-I-NHZ '+ peptides: cozn HS-Cliz-EZH-CHZ-NHZ + peptides cozn HS-Clíz-Ctll-I-CHZ-COZH + peptides “H2 çnzcozn fls-CilzCHztlï fi + peptides NH2 * V2 §HcHcoH2c2 cnzfn conacnzcoz fl Il 'Hs-cH = cH-NHccH3 HS-CHZ-fï-I-CHZOH OH CHB l HS-CHZCH-CHZOH' 26 465 929 HS-CH2? H-CH2 OCH3 nalnz nl - C113; 1: 2 - a, caa HS-CH CH2-É-CH3 2 CH3 HS-CHZCHZNH ns-cnzcnzwa / Nan: N / Hs-cH2cH2-NH- <E s CH HS-CH CH -O-å-â fi 2 2 '3 H3 HS NHCH3 OH HS NHCCB3 27 2 HS \ 4 II HS N 1 /% \ § HS O | \ HS / N magh 465 929 '46 -5 929, HS- (CHZM / \ N n HS (CH) N / äul n _ 2 n- \; N / Nšn Hs- (cHQn-NW n Rl HS- (CHZH -ÄN-à n H HS- (Cïíznll / JS n HS- (CHZH-GK 11 - / 5 ° I-IS- (CHQn-Å-xß n N Hs- (CHQn-éš fi- NH: n / X x / Hs- (cn fl n N / Å- N32 n N (C335 / HS-CHZ-Ä SRN R1).

= H, CH 28 1.2 2.4 2-4; R1 1.2 1, 2: X = 1, 2; X = 1, 1, 1, 1, 3.

= AND CH 0, 0, 0, 0, 0, 2 3 NH NH NH NH NH NH J. ns-cnz-Z ENR -CH -N \ L -_ / X HS-CH2 2 Hs-CHZÖ N H54! NRI N / \ (cnz) m us- (cnfn Ä H 2 ns-cn N 2 7LR1 N 1 Hs-æZ-fm * oj RI'- 29 H, CH3 x 3 o, NH, SW Il W ll H, CH H, CH3 a, C113 1; R = H, CH3 465 929 465 929 10 15 20 25 30 35 30 N (CH Hs / \ / Im \ / \ on OH acw / xml The utility of the compounds of formulas I and IX for antineoplastic therapy is apparent from the results of in vivo screening experiments in which the compounds have been administered in varying doses to mice in which a P-388 leukemic or B16 melanomic condition has been induced.

The compounds of the present invention are believed to exhibit antibacterial activity against gram-positive and gram-negative microorganisms in a manner similar to that found for the naturally occurring mitomycins and are thus potentially useful as therapeutic agents in the treatment of bacterial infections in humans and animals. . P * 465 929 'Activity with respect to P-388 leukemia in mice Table I contains the results of laboratory experiments with female CDF1 mice, in which a tumor inoculum comprising 10 ascites cells of P-388 mouse leukemia has been implanted intraperitogeally and which were treated with different doses of either a test compound of formula I or II or with mitomycin C. The compounds were administered by intraperitoneal injection. Groups of six mice were used at each dose level and they were treated with a single dose of the compound on the day after inoculation. A group of ten saline-treated control mice was included in each experimental series. The mitomycin C-treated groups were used as a positive control. A 30-day observation period was used, noting the mean survival time in days for each group of mice and the number of survivors towards the end of the 30-day period.

The mice were weighed before treatment and again on the 6th day.

The weight change was taken as a measure of the drug's toxicity. Mice weighing 20 g each were used and a weight loss of up to about 2 g was not considered excessive. The results were determined as the percentage T / C, which is the ratio between the mean survival time of the treated group (T) and the mean survival time of the saline-treated control group (C), multiplied by 100. The saline-treated control animals usually died within 9 days. The "maximum effect" in the following table is expressed as a percentage of T / C and the dose that gives this effect. The values in parentheses are the values obtained with mitomycin C as a positive control in the same experiment. Thus, one can get an idea of the relative activity of the present substances relative to mitomycin C. A minimum effect expressed as a percentage of T / C was considered to be 125. The minimum effective dose given in the table below is the dose giving a percentage of T / C of about 125. The two values given in the column entitled "average weight change" are the 465 929 _ 32 average weight change per mouse at the maximum effective dose and at the minimum effective dose, respectively. 33 465 929 m. ° | mß-Ol “«. ~ | “H- fl l«>. = | “N.O + vü fl ubcm | mx fi> v fifi luu fi cwâøamw mo.ov mc.ov mc.O. ~ .M.«. W Awomvwm fi @ íLß & Y ~ = um ~ = u ~ = o | . ~. ~. ~ .m. @ om.n- /. \ ~ = u ~ = u «\, N N. ~ .m.«. w. @ ° ~ .- ~ o z-: U = u | . ~ .m. @. ~. @ = ~.> w ~ Hm | m «. ~ .mw ~« .w ° m.> w fi = m | m won u \ swm F UMOMMM Hßñ fl xßä 30 Om MWÛFÜWÛÃ HEÜÅÜÜH æwmlmm xw fl AMwOHPHQHÄCH H Qnmmæm w fi ma wa. mm fi mm fi .xw .Hmm U fifl nwuwm 341 465 929 ~. ~ + “v.o + m. ~ + ~ o. ~ | >. ~ + »M.o + n. ~ + ~ W.o | n.o | «~. ~ | mc fl uwcm | mxH> u flfi uuu fi cmšocww mc.cv mc.Ov m0 _ \ Q m> fi uxwmww ß fl m fifl ä P. . uxwmwm Hmš fl xmä. @. «. @ .- .m ~ m ^. @ ~ fl ~ oz wm ~ = o ~ = o | IIZ. @. «. <. W .fl m ~ ^ .w ° ~ / \\ mm ~ zu ~ = o | ~ oz o /,. ~. ~. <. °. ~ ß ~.> w ~ = o ~ = u | o \. ~ .n.v. °. ~> ~ .- ~ m = uo ~ = u ~ = u | Nmwmzoummu. ~. ~. «. @. @ ° ~. ~ @ ~ ~ = Umm ~ = u ~ uu | mon Uxä w M mm om mn ma ß fi .Nm .Ham bn flfl wnwm ^ .m-conv H H fi mnma 465 929 35 xmmumu määmm @ fl> WOW N> HUUHÜUHMQ .NUWÉHE .S00 mm fl mumwu Eom U G fl wfim mwwu. Aowmf. mow | «ow .MN .wu fl uo fi n fi ucæ .n ..Hm vw msmxmun \ .m m fi mñ fl xmš w fi Hm> U fi> www Hmm Emuw Hmß fl m Mm fl auu fifl mäo fl ww .N vx fl> mmm0Ex mM \ + øm +. H. = v b flfl umnm wow | mx fl> w fifi. m> fl u «w @» w mßmc fl z | UUH ~ MWEÛCÜÜ _.

SJK .m 323 33 ~ = z mm ~ = o ~ = u | 2 fmwm Uxä æ M Jxw .Ham uxwmmw fl mš fl xmä U flfi cmu fl m _.muHOwv H H fl wnmß 36 465 929 Activity against B16 melanoma Table It contains the results of antitumor experiments using B16 melanoma growth in mice. BDF1 mice were utilized and the tumor implant was inoculated intraperitoneally. A 60-day long-term network was used. Groups of ten mice were used for each dose dose tested and the mean survival time for each group was determined. Control animals, which had been inoculated in the same manner as the experimental animals and treated with the injection vehicle but not with any drug, showed a mean survival time of 24 days. the survival time relative to that of the control animals (% T / C) was used as a measure of efficacy and the maximum effective dose and the minimum effective dose for each test compound were determined. The lowest effective dose was defined as the dose that gave a% T / C value of 125. For each dose level, the experimental animals were treated with the test compound on days 1, 5 and 9 intravenously. 20 __ p_ B16 melanoma Compound Maximum effect Minimum effective Avg. ex. % T / C dose dose 1 weight change za 1s7 (112) 31, e (3) 3> 214 (145) 2.4 (3) <1.6 -2.4; -1.9 110 110 (112) 3.2 (3) 3.2 + 0; 5; + 0f5 30 26 152 (145) 1.6 (3) _ <1.6 ~ 0.6; -0.6 1. mg / kg body weight 2. average number of grams per day for each maximum and minimum effective dose 3. the values in parentheses refer to mitomycin C tested in the same experiment. Due to the antitumor activity observed on tumors in experimental animals, the substance of the present invention can be used to inhibit tumors in mammals. For this purpose, they are systematically administered to a mammal with a tumor in a substantially non-toxic antitumor inhibitory effective dose.

The compounds of the present invention are primarily intended for use by injection in the same manner and for the same purposes as mitomycin C. Slightly larger or smaller doses may be used depending on the particular tumor susceptibility in question. The compounds can be readily prepared as dry pharmaceutical compositions containing diluents, buffering agents, stabilizers, solubilizing agents and other ingredients which contribute to the pharmaceutical refinement. These compositions are then reconstituted with an injectable liquid medium just before use. Suitable injectable liquids include water, isotonic saline and the like.

The invention is further illustrated by the following exemplary embodiments, in which all temperature indications refer to degrees Celsius and the melting points are uncorrected. Proton nuclear magnetic resonance (1 H NMR) spectra were recorded on a Varian XL100, Joel FX-90Q or Bruker WM 260 spectrometer in either pyridine-d5 or DZI as indicated. When pyridine-ds is used as solvent, the pyridine resonance at § = 8.57 is used as internal reference, while together with D20 as solvent TSP is used as internal reference. The chemical shifts are given in 8 units and the coupling constants in Hertz. The cleavage patterns are designated as follows: s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; bs, broad signal; dd, doublet of doublet; dt, doublet of triplet. Infrared spectra were determined on either a Beckman Model 4240 spectrometer or a Nicolet SDX FT-IR spectrometer and are given in cm; 1. Ultraviolet (UV) - 465,929 10 15 20 25 30 35 38 spectra were determined on either a Cary Model 290 spectrometer or a Hewlitt Packard 8450A spectrometer equipped with a multi-diode detector. Thin layer chromatography (TLC) was performed on 0.25 mm Analtech silica gel plates GF.

Flash chromatography was performed on either Woelm neutral alumina (DCC grade) or Woelm silica gel (32-63 μm) and with the specified solvents. All evaporation of solvent was carried out under reduced pressure and at a temperature below 40 ° C. The 1-alkyl-3-aryltriazenes are a class of reagents which are known to be useful for reaction with carboxylic acids to form the corresponding lower alkyl esters. 1-Methyl-3- (4-methylphenyl) triazene can be prepared as follows: Preparation Example 1 according to E. H, White et al., Org. Syn., 48 102-195 (1968). 1-methyl-3-on fi olvlfriaze fi. 50.2 g (0.47 mol) of p-toluene are added to a 2 liter flask equipped with a 200 ml dropping funnel and an efficient stirrer and the flask is immersed in an ice-salt bath at about -100. A solution of 46.8 g (0.55 mol) of potassium nitrite in 150 l of hydrogen is introduced into the dropping funnel and a mixture of 250 g of crushed ice and 140 ml of concentrated hydrochloric acid is added to the p-toluidine with stirring. The potassium nitrite solution is added slowly with continued stirring. for 1-2 hours until a positive starch-potassium iodide test is obtained (Note 1) and the mixture is stirred for a further 1 hour to ensure the conversion of all toluidine.

The solution of the petoluenediazonium chloride is then adjusted to pH 6.8-7.2 at 0 DEG C. with a cold concentrated aqueous solution of sodium carbonate, the color of the solution turning red to orange and a small amount of red material precipitating.

The cold neutral solution is then transferred to a dropping funnel and slowly added to a vigorously stirred mixture of 150 g of sodium carbonate, 300 ml of a 30-35% aqueous solution of methylamine (Note 2). and 100 g of crushed ice in a 3 liter flask. The reaction mixture is kept at about -100 during the addition, which requires about 45 minutes (Note 3). The solution is extracted with three 1 liter portions of ether. The ether extracts are dried over anhydrous sodium sulfate and evaporated on a rotary evaporator at room temperature to give 65 g of crude 1-methyl-3-p-tolyltriazene (Note 4). This is introduced into a water-cooled sublimer and the triazene is sublimed at 500 (1 mm); 43.3 g (0.29 mol; 62%) of a yellow crystalline sublimate with m.p. 77-800 (Note 5). The sublimate can be recrystallized from hexane to give the triazene as white needles, m.p. 80.5-81.50. It is more suitable to dissolve it in a minimum amount of ether and dilute the solution with two volumes of hexane and cool to 0 DEG C. to obtain plates of a light yellow hue and m.p. 79-810. The yield of pure triazene is 33-37 g (47-53%) (Note 6).

NOTES 1. The individual tests with starch-potassium iodide paper should be performed 1-2 minutes after stopping the addition of potassium nitrate. A 40% aqueous solution of methylamine can be used. The reaction is complete when a drop of solution no longer gives a red color with a solution of β-naphthol in an aqueous solution of sodium carbonate. The main impurity is 1,5-di-p-tolyl-3-methyl-1,4-pentazadiene (m.p. 1480). This can be removed by fractional crystallization but it is easier to sublimate the triazene from the reaction mixture. 5. The sublimate contains traces of 1,3-di-p-tolyltriazen, as can be ascertained by thin layer chromatography. Crystallization gives the pure 1-methyl-3-p-tolyltriazenene. 6. This procedure can only be performed with water-soluble amines. The procedure of Preparative Example 2 below is more suitable for the preparation of triazenes from water-insoluble amines.

Preparation Example 2 according to E. H. White et al., Tetrahederon Letters No. 21, p. 761 (1961). 1-n-butyl-3-p-chlorophenyltriazene. A solution of 2.87 g (10.1 mmol) of p-chlorobenzenediazonium hexafluorophosphate (recrystallized from acetone-methanol) in dimethylformamide (dimethylamine free) was slowly added to the stirred mixture of 0.73 g (10.0 mmol) of n-butylamine. 15 g of powdered sodium carbonate and 30 ml of dimethylformamide and the mixture was stirred and kept at -50. The diazonium salt solution can be used at room temperature; however, a purer product is usually obtained if the diazonium salt solution is prepared in and delivered from a cooled separatory funnel, which is kept at about -50 °. The mixture was heated to 0 ° C and stirred until a negative test was obtained with 2-naphthol (only a few minutes are usually required). Ether was added, the mixture was filtered and the filtrate was washed thoroughly with water and then dried. (The triazene can be isolated at this stage and recrystallized from pentane at low temperatures.

Preparation Example 3. 7-Hydroxy-9α-methokimitosane. 2.2 g (6.6 mmol) of mitomycin C were dissolved in 140 ml of 0.1N methanolic sodium hydroxide (50%) and the reaction mixture was stirred for 30 hours at room temperature. The solution is then adjusted to a pH of about 4.0 with 1N hydrochloric acid and extracted with 4 x 500 ml of ethyl acetate. The combined ethyl acetate extracts were dried over sodium sulfate and evaporated under reduced pressure at about 30-35 ° C to give a solid residue, which on dissolution in ether and treatment with an excess of 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%). 1 H NMR (pyran-gs, 3) = 2, os (s, sn), 2.14 (bs, 1H), z, 74 (bs, 1H), 3.13 (a, 1H), 3, z4 ( s, sn), 3, ss (a, 1H), 4, oo (aa, 1H), 4.37 (a, 1H), s, os (t, 1n>, s, 4o, s, 9o Preparation Example 4 Mitomycin A. 100 mg (0.30 mmol) of 7-hydroxy-9α-methoxymitosan and 100 mg (0.67 mmol) of 3-methyl-1-p-tolyltriazene were dissolved in 2 ml of methylene chloride and 10 ml of diethyl ether.

After gentle inlet boiling for 6 hours, the solution was stirred for 18 hours at room temperature. TLC (methylene chloride: methanol 90:10) revealed the presence of a dark purple spot at Rf = 0.36 more than trace amount of impurity at Rf = 0.41. The reaction mixture was concentrated to dryness and chromatographed on Woelm neutral alumina using methylene chloride and methylene chloride: methanol (30: 1) as eluting solvent. The fractions containing the component at RF = 0.36 were combined 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 °).

Analysis calculated for C 16 H 19 N 3 O 6: C, 54.96; H, 5.44: N, 12.02 Found: C, 53.96; H, 5.37; N, 11.99. 465 929 10 15 20 25 30 35 42 IR (KBr): rmax, cm-1: 3400, 3300, 2950, 1700, 1575, 1200, 1060.

1 H NMR (Pyridine-g 5.8): 1.82 (s, 3H), 2.74 (dd, 1H), 3.12 (d, 1H), 3.24 (s, 3H), 3.54 ( dd, 1H), 3.96 (s, 3H), 4.22 (d, 1H), 4.84 (bs, 2H), 5.02 (t, 1H), 5.38 (dd, 1H). 1630, 1 The yield in this example can be increased to 63% if methylene chloride is used as the reaction solvent and at room temperature for a period of 24 hours.

Preparation Example 5 In a 250 ml single neck round bottom flask was introduced solid sodium carbonate, a 35% solution of amine (amount as in Preparation Example 1) and ice and the suspension was stirred at -SOC (ice-salt bath). To this suspension was added dropwise a cold suspension of p-chlorobenzene diazonium hexafluorophosphate (Aldrich Chemical Co.) in ice, water and sodium carbonate (solution about pH 7). After complete addition, the reaction mixture was extracted with diethyl ether. The combined diethyl ether extracts were washed again with water, dried over sodium sulfate and concentrated. The yellowish solid residue was purified by column chromatography on Woelm alumina using hexane-methylene chloride (1: 1) as eluent (1 H NMR recorded).

Egèmgél 1-10 The triazenes 1-7 of Table III below were prepared according to Preparative Example 1 above, exemplifying the triazene of Example 1. The triazenes were purified by column chromatography on Woelm alumina.

The triazenes 8-10 in Table III below were prepared according to Preparative Example 5 above. n NN: uouzzwzu: omm 465 929 43 = u ~ = u = z »zuz <= wun = u | a ÛNzuwwzu ~ zuxz | znz <= won = u | m .. ~ = z ~ = o ~ = omm ~. .u ~ = u = zuo ~ = u ~: z ~ = u ~: um ~ = o | Aïl% 1v uz ^ í %% xvr ~ = u ~ = u = z | z | zv = øun = u »m 1 ~ = z ~ = u ~ = u «^ É% ¶% vm O: ~: u ~: u: z | z | z <= oun = u..n ._ ~ ._z ~: u ~ = u | zo ._ / IK / | \ ~ _ ~ = u. = u = z »= | z- = wun = um = ~ = z = u ~. ~ = o. n. n = u ~. ~ = u.4 | znz ~ = won = u | m = ~ = z ~. ~ = u. ~ = u ~ = ~ = u: z | znzv = wun = o | m | «u + ~ z | ^ ï1ïLv | ~ = u ~ = zn = u.

HHÜNNHHÉ. ÖÉÖWMHÜHAHÛNÜHQ. .Hg fi mwaw fifi wvmm ~ ~ z = z | znz x Al |||||||||||||| @ <@ zx +: zu Hw fl wNm fi HB HHH HHGQME 44 465 929 Q c ~ = u |: z..z ._. ze: @u «u | m .. ~ = z ~: u \ ~ ow S o, ~ ww ~ ~ o T: u = u = z | zuz _. 03 .... ._: z: UÄ i m \ o O. n = uo ~: u ~: u = z | zuz ~: @ u ~ u | m nwå + ~ z åo ~ = z ~ = u ~ = uon: u _. : www-BB u fi mmëø fl conm fl n CÉE u: .Awdwwnwxnnl Am @ Howv HHH dammas 10 15 20 25 30 35 45 465 929 'Example 11 1- [2- (3-nitro-2-pyridylthio) ethyl] -3- (4-methylphenyl Triazene A solution of 4-methylphenyldiazonium chloride was prepared as described in Preparation Example 1 starting from p-toluene and adjusted to pH e, a-7.2 at 0 ° as described in this Preparation Example. A solution containing 21.15 mmol of the diazonium salt in 45 ml of solution was prepared in this way and introduced into a dropping funnel, which was connected to a 250 ml three-necked round bottom flask containing 5.34 g (20.0 mmol) of 2 - (3-nitro-2-pyridylthio) -ethylamine, 7 g of sodium carbonate and 150 ml of dioxane, which had been added to the flask in that order. 6 ml of a saturated aqueous solution of sodium carbonate and 10 g were added to the flask. The flask was cooled in an ice bath and the contents were stirred mechanically. The diazonium salt solution was then added dropwise over 1 hour from the dropping funnel. After complete addition, the reaction mixture was allowed to warm to room temperature and then extracted with 3 x 400 ml of ether. Drying and evaporation of the extracts gave the desired product, which was purified by chromatography using an alumina-filled column 2.5 cm in diameter and 25 cm in length and using hexane-methylene chloride (4: 1). , hexane methylene chloride (3: 2), hexane: methylene chloride (1: 4) and finally methylene chloride containing 1% methanol to develop and elute the column. The appropriate fractions in question (identified by TLC) were combined and evaporated to give 2.5 g of the title compound.

Example 2 Preparation of 7-alkoxy-9α-methoxymitosanes (12-19) A solution of 2.4 equivalents of triazene in methylene chloride 465 929 46 46 methanol (4: 1) was added to a solution of 7-hydroxy- 9a-methoxymitosane (prepared according to Preparative Example 3) in methylene chloride / methanol (4: 1). The reaction mixture was stirred at room temperature and the course of the reaction was followed (10% methanol in methylene chloride). The 7-alkoxy-9a-methoximitosan product was obtained by thin layer chromatography (TLC) and appeared as a dark purple spot on TLC. The reaction mixture was chromatographed on Woelm alumina when the reaction was judged to be complete on the basis of TLC and the 7-alkoxy-9a-methoxymitosane was obtained as an amorphous solid. The products prepared were identified as Examples 12-19 in Table IV below. 47 465 929 .. = ~ .uø. ««. M .. = ~ .s. °° .m ._ = ~ .u. «~ .- ..: ~ .øo. = Ov ._ = ~. u.wm.n .. = m .mv ~ .n _ ._ = ~ .un.w ~ .m ..: ~ .un.w>. ~ .. = ~ .mn.ø ~. ~ 1 | _ .. = n ^ m. = w. ~ .. = ~ .u. @ ~. ~ .. = ~ .u. ° ~. ~ ~ .n = u. = u | nm nn. .. = ~ .mn. «@.> .. = ~ .uuq -«. m .. = ~. ». <° .m .. = ~ .em ~.« .. = ~ .mu. ° e . «.. = ~ .u. ~ M. ~ .. = ~ .m. = ~. ~ Ø = .e ~ .z“ m ~ .w. = N -. @ M .u nu .. = ~ .mn. ~ = .n .. = ~ .un .-. ~ .. = ~ .mn. ==. ~ <~ .e ~ .z “- <. @ _:“ e @. @ m .u ua 1 .. = n .a. «@. ~ .. = - .a. ~ f. ~ ._ = m.». = @. = @ = u ~ = u ~: u ~ = u «uz n ~. ann un mä fiflfifl umv fl wzåw fi u fIEO. zu 2 .mlx .Åman-amy: Zz: d HwEHOh Hwn fifi wv fi m o M \: u oz O HUGMWOUHEMNOHÜEIÜGIHXOMHMIN> H H fl w fl d fi \ | .. \ .. 48 .. = ~ .uo. ° wm .. = H .u .. <.. = ~ .u. <~. <mwo fl. = fi- .. = ~ .uu. @@. fi .. = ~. ». w>. ~ .. =. .ø. ~ m. ~ .m ~ w ~. =.> ~ .. = n. «.« ~. ~ ._ = m .s. @ °. ~ ._ =. _nn. @>. ~ 1. = ~ @ ~ .øemn .. = ~ .un ~ f ~. ~ .. = ~. @. «°. ~ .. = n. @. = @ .. n = uou = z ~ = u ~ = u »ww ~ = u ~ = u || m ß .mn.w ~.» ..s.w «.> ._ = ~ .vu. @ ~ .m 465 929.» ». @ .m www. .. = H. ».-. W .. = ~ .e. = @ .- .. = ~. @. @ ~ .- .n @ .ø .z .ww.m. =“ W ~. @ m .u nu. = _-. = ~ w ~ .. =. .øu. <°. ~ .. = ~. =. @@. n .. = ~ .u. @ m. ~ = w. @ .u .mmm fi. = fl @ ~ .. = n .u. @ ~ .n .. = ~. @ n. @ ~ .n .. = ~. ». <@. ~ @ w. @ .z _ =». m _: “» fl. @ m .u “e. = @ -. ° -M .. = ~ .nn. @>. ~ ._ = ~. @ P. ~ .. ~ .. = n. @. ~ @. ~ A¶s <@ Yw = um ~ = u ~ = u | um ø fl = w ° ~ .. = ~ .uo. @ nm ._e. ~ @ .-. ° .- .m ~ w ~ .. = ~ .u. @ ~. ~ .. = ~ .øu. ~ «.n .. = ~ _v. ° m. ~ nø. ~. .z .wø.m _: ~ va. @ m .u nu .m.> «.ønau .. = n.». e ~ .n .. = ~ .u.ø ~ .n .. = ~. nn. = and ~>. ~. .z. «~ .m _:.« n. @ m .u nu .eonn .ønfn .. = _. @ n. ~>. ~ .. = ~ _-n. @ «. ~ .. = @ _n_@w.~ ^ Xï¶Av1 ~ = u ~ = u || ø m ~ | .z ._ = ~ .uu. «-. m ..- u ~ = u mme fl .. = ~ .u. ~ ° .m .. = ~ .e. = W. ~ .. = ~ .w. @ ~. <Wø fi oš mo Mmm umuwmäuhox o.- .eww fl ..: fl .wø.øe.v ..: ~ .E . @ wn .. = ~ .u.vm.n ~ <.. ~ _:. = e. @. = .mß. ~ m .o .m .m ~> «. = <@ ~ .. = @ . =. -. ~ .. = H .ø. @ ~ .N .. = ~. @ N. @>. ~ \ | 4; / _ <._. .z _m «.m. = _ = w. ~ m .o ua .ewwm .øwvn .. = ~ .u. @ m. ~ .. = <.e. = -. ~ .. = ~ .u. @ m. ~ oz | ~ = u ~ = u || æ vu-Å Emm .HQ mmh fi mcmumucwšw fl m Jaså: H. @. m || .äuqu? = m: z .: Hmšnom ammïwxm Aw # HOmv> H HAQQMH 49 465 929 .u fl o .OOH ..Hm uw m3mxmuD ~ <c fl oæëoß fl ñ> m mæ fi ønox fl m Eöcwm Hmnaamuwëmnw cmë fl AU mzHu | ^ |. : Oo | ^ +. fl umnm> HmwnO som Hmaæxw fi oâ AU Aoæmfv moæ | woæ .mm ~ mo fi uO fl Q fl uG <.h ..am um mšmxmub An ßwåäw nm .Hšm fi wuowp ua E ..: ~ .mn. ~ m.> cwc ~. ~. .vo.w ~ .m .. = ~ .ew ~ .m .. = ~. ». @@. ~ .m ~ @ ~. = -_ _. = ~ .a.ow.- .. = ~ .u. ° ~ .v ._ = ~ _uv. <@. n vß.ø _:. @ ~ .m _:. «v.nm .u _». ° w @ ~. ° = - ..: ~.:. «@. ~ .. = ~ .øu. ~ Mn .. = ~. @. ° ~ .n of ~ n.« .Z. = Mm _: hm fi.- m .u na _ ° @ ~ n. = @ «~ .. = H .u. @ ° .n .. = ~ .s .-. ~ .. = ~.«. ~ @ .. @@ ø \ = u | ~ = u | um »_ mwø fi .. = ~ .ø¶. °« .m .. = ~. ».« =. m ._ = ~ .s. @ m. «. . ° = - .omw fl .. = ~ .u. ° ~. «.. = ~ .uø. @@. N .. = ~ .s. ~ M. ~ -. =. .z .m ~ .m. = ~ m «. ~ m .u nu .øøß fi ~ ° mm ~ .. = n .w.« ~ .n .. = ~ .n. ° ~. ~ .. =. .w. = °. ~. øu.ø ~ .z _ @@. m _: “wa .. ~ .. = ~ .mn. °°. ~ .. = n .m. @@. ~ ~ = uo ~: u ~ = u | | z nu.

Emm .HG mw fi mcmwmwsmqw fi m. ~ | Su. z fl .Q .mw |. = fl u fi ~> m. mzz = ~ Hmmswxm ^ .muHOmv> H a fi mnm fi 465-929 10 15 20 25 30 35 50 Example 20 9a-methoxy-7- [2- (3-nitro-2-pyridyldition) ethoxyfmitosan (20) 580 mg (1.73 mmol) 7-hydroxy-9a-methoximitosane was introduced into a round bottom flask and dissolved in 60 ml of methylene chloride.

The triazene of Example 11, about 2.5 g (5.7 mmol), was added to the solution in the flask and the mixture was stirred for 14 hours at SOC and then 8 hours at room temperature.

The course of the reaction was monitored by silica TLC using methylene chloride: methanol (9: 1). The reaction mixture was kept at room temperature for a further 26 hours and then worked up by column chromatography on a column with a diameter of 85 mm and a length of 30.5 cm and which was filled with alumina.

The solvents used sequentially for development and elution were 200 ml portions each 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 calculated for C 22 H 23 N 5 O 8 S 2: C, 45.65; H, 4.09; (corrected for 0.5 mol% CH 2 Cl 2): N, 11.82 Found: C, 45.74; H, 4.14; N, 11.61 IR (KBr), Vmax, cm-1: 3440-3200, 3060, 2930, 1720, 1570, 1510, 1395, 1335, 1210, 1055. 111. mm (pyran-gs, 8): 1 , s1 (s, an), 2.0 (bs, 1H), 2.61 (bs, 1H), 2.98 (bs, 1H), 3.08 (s, 3H), 3.20 (m, 2H), 3.39 (d, 1H), 3.83 (dd, 1H), 4.07 (d, 1H), 4.59- 4.89 (m, 3H), 5.21 (dd, 1H) ), 7.16 (dd, 1H), 8.31 (dd, 1H), 8.71 (dd, 1H). By applying the procedures of Examples 11 and 20 to andraul (3-nitro-2-pyridyldithio) alkylamines having 2-6 carbon atoms in the alkyl group, mitosane derivatives of the following formula can be prepared.

N \ ss- (C112) n-O / ° 2 ca3 D = 2-6 1 _ R - H VPÄIGI 'C1-ö- = 2 * 6 N R1 = H or C1-6-alkyl Examples 21-34 7-alkoxyditio- The 9a-methoximitosanes 21-34 of Table V below were prepared using General Method A or B below, as set forth in Table V. Physical data for the mitosane compounds 21-34 are set forth in Table VI below.

Method A To a deoxygenated solution of mitosan according to Example 20 (about 0.1 mmol) in 3-5 ml of acetone is added with stirring and under argon about 1.1 equivalents of triethylamine, followed by dropwise or portionwise addition of one equivalent or a mercaptan ( in the case where the starting mercaptan is impure, more than one equivalent of thiol) in 1-2 ml of acetone is required.

In most reactions (in cases where the starting mitosan of Example 20 and the product have very close R values at TLC, monitoring by high pressure liquid chromatography (HPLC; | LBondapak-C18 column) is used) the course of the reaction is monitored by silica gel thin layer chromatography (10% methanol in methylene chloride). 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 stage, the reaction mixture is concentrated under reduced pressure (at about 30 ° C) and the residue is chromatographed on a neutral Woelm alumina column (6.4 mm x 25.4 cm), which has been filled by slurry with 2- 5% methanol in methylene chloride. This procedure separates the desired mitosan product from the pyridyl thione by-product retained on the column. The product thus eluted using 2-5% methanol in methylene chloride is further thoroughly purified by silica gel flash chromatography using 5-7% methanol in methylene chloride as eluting solvent.

The main band corresponding to the product is isolated and the amorphous 7-alkoxydithio-9a-methoxymitosane is characterized.

To a solution of about 0.1 mmol of mitosan according to Example 20 in 2-5% acetone (methylene chloride may also be used but acetone is preferred) in 10 ml of methanol is added about 6 drops of a saturated aqueous solution of sodium bicarbonate (in the case where mercaptan is L-cysteine, this base is not used), followed by the addition of one equivalent of mercaptan in 1 ml of methanol (water is used if the starting thiol is water soluble). The course of the reaction is monitored by TLC (silica gel, 10% methanol in methylene chloride). After completion of the reaction, the reaction mixture is diluted with 15 ml of water and concentrated to about 10 ml under reduced pressure at about 30 ° C. The resulting solution is chromatographed on a C-18 reverse phase column with stepwise gradient elution (100% water to 80% methanol in water; elution with water separates the yellow pyridyl ion product from the product retained on the column). The product, which elutes as a red headband, is recovered and concentrated to give 7-alkoxydithio-9a-methoxymitosane as an amorphous solid. If further purification is required, the above chromatography steps are repeated. 465 929¶4 S4 Table V, 7-alkoxydithio-9a-methoximitosanes à or Rss / \ v / 0 asu + g_0_ ---->, B CH3 Example Thiol no (353) Method Product _ 'H' 21 ethyl-2 -metcapto- AR = -CH 2 CH 2 OCH 3 acetate 22 _3-mercapto-1,2-AR = -CH 2 CH (OH) CH 2 OH propanz _ - + 23 3-mer aptoproplon B = -CH 2 CH 2 COO Na acid 2 4 cysteix; B R = -cH2cH (Nnf) coo 'thiophenol * A R: Gus 28' p-aminobenzenethiol A R; '<::::> “NH2 W ll 26 p-nitrobenzenethiol A 27 p-methoxybenzenethiol AR 5 * 465 929 Tåbell V (cont.) Exemel Thj-Ol nr (RSH) mietod Product coo'Na * 29 Z-mercaptobenzoic acid B 'R = 30 Z-Nizro-β1-Gnergapto- BR = NO2 benzoic acid _ + C00 Na 31 4-Mercaptopyridine AR = ~ <í: ïSš N 32 2-Mercaptomethyl-1- AR = -CH2- <ï: Än methylimidazole NI CH3 * fä §HcocH2cH2cHcoo'Na 33 glutathione BR = -cæ12 <|: H coNHcH2coo'Na + 3 4 dimethylamine- BR = -CH2CH2N (C33) 2 ethanethiol 56 465 = 929 .Omc fi .mwø fl .Ocm nnm. .owmn .. ~ = m. «. ~. ~ H. = ~ .øø.m ~ .m .. ~ = @. =«. = ~. ». ~ m. <.. = ~ .a.wm . «._ ~ =>. @. = ~ .U.

-. «.. ~ = H .-. = H.v.m °.« .. ~ = m. «. ~. ~ H. = H.uu.« W.m .. ~ mH .-. = ~ .U. > mm. = nw> =. ~ .. = ~ .mn.mm. ~ .. ~ = ~ .w. = ~ .u.wm. ~ .. ~ =>. w. = fl. = «> ~ wH ~. = ~. ».> @. ~ ..mH. @ n. @ m. ~ .. ~ = m.>. = ~ @ ~. ° ~ mn. ° ~ mm.« -. ° ~ m . = fi. ».> m. ~ ..m ~ _m.« @. ~ .. = mmwß.H m = umo ~ = u ~ mu | nm H ~ .ansa .x ~ s>. .ac 1xøe <. .Emm. @ .. Hwsnom Mc. ~ M =. m fi. = on = o. > = w.mm;, = fl u «u> m. «» ~ W «mzz = ~ Hwlawx fl zu. .HGGMWOHAE fl NOUQEIN QIOHUÉHÜHXOMHMIN.

H> .HHwnmw 465 929 57 .mwc fi. ° - ~. = - ~. = ° - .e «m ~ .m °« ~ .am «~ .m> m ~ .ø ~ w«. ° mw ~. m ~> ~ .o ~ a ~ .om «m ..: ~ .e.« w.> .. ~ m @. ° ~ .H. «_ m ~ .uw.o ~ .m _.:Hs« @. «.. = ~ .E.mm.« .. ~ = m. ~ H. = Fi. W. @ =. «.. ~ ==. HH. °.«. = H.oo. ~ wm .. ~ = <. ~ «. = ~ .U.wm.m .. ~ =«.>. = ~ .U.> ~ .N ø fl m .. = ~ .m. ° ø.m .. = m_a . «M. ~ .. ~ = v.» .m-.m ~ fl. = ~. ~. @>. ~ ..: H.wn.ow. ~ _. = m.m. >>. ~ + øz | ooo ~ = u ~ = o | um m ~. .. = ~ .am> .. = ~ .mn.mw.w _. = H.mn.m ~ .w..m ~ .uw.m ~ .m .. ~ = @. ° H. ». = H. ~ m. «.. = Hs« @. «.. = fl. E.> Mv __ = H.mn. @ ~.« _. ~ = «.-. = Hw @ °.« ._ = ~. ° ~ ° ~ .mn. «Mm. ~ = ~ .H ~ .m.«. M ~ .oø. «@. M. => = ~ .MH-.m ° ~ H _. ~ M «. ~ H. = Ho> mm. ~ = ~. ~ Hm«. = H. = «W ~ .o ~ v ~ .mm« ~ .wø.æ ~. ~ .. = HsmH.m _. = nmß =. ~. ° @ m ~ .mmw ~ .mm @ ~ c ~ m _. = ~ .ao ° .n .. ~ = m. @. = ~. ». mo.m. = ~> ~ .m @ m ~. = «« ~. «-. m- _. = H.mn.o @. ~ ._: H.mn.m« .H..mm.w. ~>. ~ mo ~ mu = u ~ = u | nm - ä .HE.-E>. _E = .x ~ s «. .Emm. @. Hwawom Mg _umz. mw. = omzo. > = m. W:: «ø« u> m. mums mzz = Hwaawxm ~ ^ .wuuomv> H fifl wnma 58 '465 929 .mmøn fi ä fifl Äzm fi .ocm fi .mmm fi- omv fi ._ ~ = ~ .ß. = ~ .ø. ° m.> _. ~ = ~.>. ~. >. = ~ .øw.m ~.> .. ~ = ~.> .: Hw

HH. ~ ._ ~ ==. HH.m. «. = Fi. Øu.m ~ .m .. = fi_ a. ~ m. «.. = Hs> m.« .. = Hsm «.« .. ~ = «. ~ H. = ~ .uw °.« .. ~ = °. ~ fi. m. «. = ~ .uu. ~ @. @ .m> m ~ .ø ~ w ~. = mwH qwm _. = ~ .um ~ .m .. = nmw = .n .. = ma @ m, ~ .m ~> ~ . ° ~ «~. =« «~.« ~ Mw ° ~ .. = ~ .mn. ~ W. ~ .. = ~ .mn.mm. ~ .. = mmm> .H um m ~ .. ~ m @. ° Hwv = H_uu. . = @ ø ~. = H- @w. «.. = ~ .e. @@.« .. ~ = m. ° ~. = ~. ». ° ~. #. °° n ~ .ø« m ~. ° m <~ .. ~ = ~. @. o. «. = H.ou. = fi.« _. ~ = m ~. = ~ .u. .mmv ~ .m> mH.mmw ~ @ ~ m.H @ m ~ =. «.. ~ = ø.m ~. °.«. = H.uu.wn.m .. = n.m. .m ~> ~ .e ~ «~ .øm« n. «~ nw- m ~ .m .. = msmH.m | mm. ~ \. = nm ~ mH = oou = u ~ = uum« ~ _ Nm ».H42 .x ~ s>. .E = .x ~: <. .aan. @. Hwagou M:. ~ Mz. z fi _ = o ~ = u. > = m. ml = fl @ fi ~> @. ßu fl w mzz = Hw fl swxm a A. muïuOwl H> HHQQMB '465 929 59. = ~ ° ~ .mmc ~ .ø ° -. => -. o- ~ .m «~ H.mm-.omm ~. °° < ~. ° vv ~ .om «~ .o> m ~ .m ~ wH.m ~> ~. ° m @ ~. ° om @ .o @« m .. ~ =>. @. = ~ .U. ° m.> .. ~ =>. W. = ~ .Ø “« @. @ .. ~ = ° .HH. ~. «. = H.øuV> ~ .m_. ~ N @ .oH _. = ~ .u.mm.v..m ~ .a.mm. «.. ~ = m.-. = ~ .n.wo. <.. ~ = o. fi H. ~.«. = ~. @ ø . «@. M _. = Nm fl m.m .. ~ = o. ~ H. = H.ø.> M. ~ .. = ~ .w. @ °. ~ .. = ~.». ~ O .m _. = ~ .mn.o °. ~ wm ~.-m .. = ~ .mn.mw. ~ .. = H_e.oo. ~ .. = mm-, H mnuo | um »N .. ~: w.w. = ~ .ø. ~ o. @ .. ~ z @. @. = ~ .ø.mm. ~ ._ ~ = Q.HH. ~. «, mm = ~ .m = H ~ .oH-. = ~ .uu. ° ~ .m..N ==. HH. = H. ». w @.« .. = ~ .mm-.mmm ~ .o = «~ .e.mm.« .. ~: ~ .-. = Hu ~ °. «.. ~ m °. ~ Fl .mv« ~ .m = m ~. ° ~ m ~. ~. «. = ~ .Øø.m> .n .. ~: H. ~ H. = ~ .ø.m ~. ~. ° ~ @ ~ .m «w ~ .m- ~ .. = mm« ° .m .. = ~. ». ° om. = fi. mn.m @. ~ . ° «m ~. = M ~ mo @« m mH ~.-M .. = H.mn.> m. ~ .. = ~ .a. «MH. = Mwm>. ~ Noz um w ~ ~ fl lEU ~% @ E> ~ .EC .xdäxv -EQQ ~ exercises Hwšuow .Hz. ~ Nz. zw. = o ~ = u. > =. «. m | J = fi ø «u> m._ ~» «@ mzz: H Hwmawxm n; . 11 ... ^ .muH0wv H> H fi wnmä 60 465 929 .mNcn.oßO fi ~ O fififl .m fl Nn.mOm fl.0 vm ~ .omm ~ .mmc ~ .mmm fi .. ~ z>.>. = ~ .Ø.> «. w _. ~ = fl. @. = fi. wm ~. @ ._ = ~ .mn.m. ~ .. ~ = v.>. @.>. = H.øø .-.> .. ~ = <.ß. = «.». = °.> .. ~ = «. ° H_ ~.«. = ~ .oø. @ Hm _. = Hs> m. «.. m ~ .e.mm.« ._ ~ = <. ~ H. = Fi. Um =. «.. ~: =. H ~. ~. @ .: fl. Uu.o @ .m .. ~ = @ .-. = H. @ . @ mm. = mm @ °. ~ ßz OOU. ° @ w ~ .mww ~ .o ~> ~ «~ m. = ~ @ .. = ~ .mn.mm. ~ .. ~ m ~. @. = ~. ». = M. ~ + | . ° ~ «~. ° = mn.omnm .w« ~ .v- .. = H.wn.mm. ~ .. = fl. S.mm. ~ .. = mmw> .H um @ ~ .. ~ = ~. @. = ~ .ø.> m.> .. ~ = ~. @. = ~ .ø. ~ ß.w .. = H.mn.vw.m .. ~ ==. H ~ . ~. «.: ~. @ W. ~« .M .. ~ mw. ° ~. = ~. ». .m @ ° H.mo- wm. «.. = ~ .s.mm.« .. ~ = m .-. = ~ .u. .ø- ~. == -. mn- «°.« .. ~ = ° .H ~. ~. «. = ~ .øu.« @. m. = H «~. ° m« ~ .mm « ~ .. ~ = m. ~ H. = H_w.> @. m ._ = mw »° .m .m> m~.°m@~.o~> ~ .. = n_s.~Hm|@m . ~ .. = ~ .mn.ow. ~. ° ~ m ~ .c> -. Qm «m ~ m ~. @ - _. ~ = ww = Hum °. ~ .. = ~ .m.«> .H ~ = z um @ ~ _ .B: sei ... a .äeá ... aFš fl ëæä ä nl. m I Hwmšmxm .uma. : H .mo zu. > = w. @ |.: «ø fi u> m. mumw mzz: H A.wuu0uv H> H fi wnmä 465 929 61 .mwc fl .Q fl m fl .ocm fl -mmm fi .mo fifi .om-Ä .. ~ = m. «. mH = ~. @ ø. @«. @ .. = ~ .ew> ._ ~ = m. «. m. ~. = ~ .øu.mm.> .. ~ = m.oH. ~ .v. = ~ .øø.m ~ .m _. = ~ .mn . ° m. «.. m ~ .s. ~ M;« .. ~ = «. ~ H. =«. U.> °. «_. ~ = O.HH. ~ .v. = Fi. Uu . ~ wm _. ~ = m.HH. = H.ø.mm.m .. = mm @ om .m> m ~ .o ~ @ ~. ° m @ ~ -mm ~ m _. = HaoQ. m .. ~ = fi. w. = ~. ». ~ = .m..mä 11 .ø ~> ~ .ø ~ m ~. ° m« m. ~ v ~ .m- .wn. = w. ~ ..: fi. wn.m @ .H..m. = mw> .H z /. \ ua än ..z ~ .mn. «H. @ .. N = n. @. = ~ .w. m @.> .. ~ = m.m. = ~ .ø.m «.> .. ~ = ~. ° ~. ~ .v.m ~ .ww.H ~ .m .. ~ = m.o ~. = ~ .u. => = ~ .mH @w. «_. = ~ .a.mm.« .. ~ mH .-. = fl. u.wo. «.ovm ~. ° =« fi. ° m «~ _. ~ = mQ ~ .m. «. = H.øv. ~ @ .m .. ~ = @ .- az oou. = ~ m ~. =« m ~. = ~ w ~ ° ~ mo ~ m. = ~ .ø.mm.m _. = ~ .s. ° m. ~ .. ~ = mw + 1. ° - ~. ° ~ @ ~ .ø «« ~ .H ~ mm-.: ~. ». mm. ~ ..mH.mn.wm. ~ .. = ~ .mm> »~ ~ oz um om ..- E .x ~ e>. .e = .x ~ e <. .Emm. @. HQEMQM Hg .um =. mH = om = o. > o w. mf c fl ø fi uæm. mama mzz m Hwmawxm fl TmQHOwJ H> HHwQmB 62 465 929 .omø fl .møø fi .mean .o fi N fi .oøm fl .ovm fi .mouå .mme fl .emma. ..N:>. O..m.v. =. .øø.m @. «.. = ~ .e.w«. «.. ~ = m. = ~. = H.». om .... ~ = H.m ~ .z ~ .w. ~ °. «.. = .. e. cm. ~ | °>. ~ .. ~: m. ° H. «.«. = ~. @ u. °> .m ..N = H. ~ .. = H.ø. ~ wm. ~ = H. <.. w. «. = .. uu. @ ~ .M .. = mm@~.m._=me .m ~ w ~. = m @ ~ .m ~ »H« ~ m oH.m | m @. ~ .. ~ m ~ .wm ~. = ~ .uu.mm. ~ + wz | ooo ~: u = zow. . ° ~ «~.°m~mo@mm .w @ mw ~ n .. ~ = ~.« H. «.>. = ~ .Wu. @ H. ~ .. = @. M.Hm. Fi muwmunnm mm _ + ßz | oommu ~ = u ~: uou = L i .. = ~ .v.mm.w .. ~ = m. ° H. «. <. = H.øu.m ~ .m .. = .. e.mw. «.mwc fi .. = ~ .e. @«. «.. = ~ .m.HH.« .. ~ = m .-. = H .m ° -. = - ~. °° m ~ .ø.w =. «.. ~ m °. Fl H.«. «. = .. øu.« Wm »m - .. ° m« ~. ° m «. _. = ~ .mm «. ~ _. = ~ .s. °«. @ .. nn ~ = u .ø @ m ~. = m @ .. ° mw ~ cwm .w. @ ø.m .. ~ = ~. «. M ~ .w. @ M. ~ ._ ~ =«. Ww. ° ~> ~. = Wm ~ .envm .n ~ n. @ -. = ~. ». N>. ~ .. = H.mn.mm. ~ .. = ~ .m. «>. ~. 21 »v | ~ = u | um - z: Ja-U .Kmëâ 1 :: .UBEÉ Ašmm. .av HmÉHOw u: .unz. nu. = om = o. > = m. mf c fi w fl uæm. øumw mzz = ~ Hwmamxm ..m @ ~ o «. H> HHw @ fl a_ 465 929 63 .MM S00 WN C®% Qs ~ ®vm® PÜHHCUÜ .HÜQÜWOPHE .HÛH WÜÜGWïKÄÜ ÛNQ »ß. ~ = No ~ \ m.«. = H.øu. @ ~ .m .. = ~ .a.> m. «.. ~ = m .-. = Hu wo.v..N = ~ .cH.m. «. = ~ .uø. <@. ~ | .. = fi. a. = «. m .. = mm @ =. m .. = m .e.mo.m | ° @. ~ .. = ms ~ w. ~ .. = @ .m.« ~. ~ .. = H.mn.mm.H .. = mm >>. Fi ~ .m = oz ~ = o ~ = u | um «n ~ | E .xmE>. .And .XmE .Hmx. mH _mom = u. >: m. m |. = fi o «u> m. mumø mzz = fl wmemxm A TmMHOwC H> H fl mnmå.

Claims (20)

465 929 10 15 20 25 30 35 PATENTKRAV êë Compounds of formula II or formula III α-0 ca ocna 'R3-Aik -ss-Aik 2 2 - 1 2 II ca -al R4-SS-Alk2- III CH wherein Alk1 is a straight or gregain chain alkylene group having 1- 6 carbon atoms when R is joined thereto via a carbon atom thereof and with 2-6 carbon atoms when R 3 is joined thereto via an oxygen or nitrogen atom thereof and R 3 and -SS- in this case are attached to different carbon atoms; Alk is a straight or branched chain alkylene group with 2. -6 carbon atoms, R 1 is hydrogen or alkyl with 1-6 chelate atoms; Q R 3 is carboxy, alkanoyloxy having 1-7 carbon atoms, di-V alkylamino having 1-12 carbon atoms, alkanoylamino having 1-7 carbon atoms, a heteroaromatic group having 1-2 rings, 3-8 ring members in each ring and 1-2 heteroatoms in each ring, which heteroatom 2. Compounds according to claim 1, (h. Vr 465 929 - consists of oxygen, nitrogen and / or sulfur, or 1,2-dihydroxyethyl; and is a heteroaromatic group having 1-2 rings, 3-8 ring members in each ring and 1-2 heteroatoms in each ring, which heteroatoms consist of oxygen, nitrogen and / or sulfur, with the proviso that the heterocyclic group is connected via a carbon atom, which is bonded to at least another carbon atom (ie the carbon atom attached to -SS- may not itself be attached to two other heteroatoms), phenyl or B-substituted phenyl or a glutathione residue, the B substituent being one or two lower al coxy, amino, carboxy or nitro groups, or R 4 and the adjacent sulfur atom together form S-cysteinyl, wherein S-cyst the einyl group may be esterified or present in salt form, and non-toxic, pharmaceutically acceptable salts thereof. / hence that Alkz is ethylene and R1 is hydrogen. Compounds according to claim 2 of formula II, characterized in that Alk is ethylene 1 and R 3 is acetylamino. k ä n n e t e c k n a d ë 465 929 “f 10, \ 15 20 25 30 35 Compounds according to claim 2 of the formula II, characterized in that Alk1 is ethylene and R3 is acetyloxy. Compounds according to claim 2 of formula II, characterized in that Alk1 is methylene and R3 is 1,2-dihydroxyethyl. Compounds according to claim 2 of formula II, characterized in that Alk1 is ethylene and R3 is carboxy or a non-toxic pharmaceutically acceptable salt thereof. Compounds according to claim 2 of formula II, characterized in that Alk1 is methylene and R3 is 1-carboxyaminomethyl or a non-toxic pharmaceutically acceptable salt thereof. Compounds according to claim 2 of the formula II, characterized in that Alk1 is methylene and R3 is 1-methylimidazol-2-yl. Compounds according to claim 2 of the formula II, characterized in that Alk1 is ethylene and R3 is dimethylamino. Compounds according to claim 2 of formula III, characterized in that R 4 is phenyl, 4-nitrophenyl, 4-methoxyphenyl, 4-aminophenyl, 2-carboxyphenyl or a non-toxic pharmaceutically acceptable salt thereof, 4-nitro-3- carboxyphenyl or a non-toxic pharmaceutically acceptable salt thereof or 4-pyridyl. The compound of claim 2 having the formula é? 465 929 'NH ao coaân O 2 zcnzconn O CH OÉNH cncnzsscxzcazo 2 2 Hozccxznnco 3 C33 5 or a non-toxic pharmaceutically acceptable salt thereof. 10 Compounds according to claim 1 of the formula Ib NO 2 O CH 'NH _ 2G // \\ SS Alk 0 2 2 1 5 __ èpCI-I3 Ib css f -al 20 wherein R 1 and Aik have the meanings given in claim 1. 2 The compound of claim 12 having the formula 25/02 ° CH O fl na ss-cszcgz -o 2 _'N -oca e 3 XVII 30 C83 14. A pharmaceutical composition characterized in that it contains a compound according to claim 1 together with a pharmaceutically acceptable solvent, diluent, adjuvant and / or carrier. 465 929 éf 10 .Û 15 20 25 30 35 Process for the preparation of a compound according to claim 1, characterized in that at least one equivalent of a triazene of formula V or formula VI is reacted Ar-N = N-A1k2-ss-A1k1-R3 Ar-N = N-A1x <2-ss-R4 V VI with one equivalent of a mitosan of the formula IV OI CH 2 OCNH 2 H 1 l wherein R 1, R 3, R 4, Alk and Alk have the meanings given in claim 1 and Ar is the organic the residue of a diazotizable aromatic amine, under reaction conditions in an inert organic solvent at a temperature from 0 to 60 ° C until a substantial amount of the product of formula II or formula III has been obtained. 16. A process according to claim 15, characterized in that the triazene of formula VI is 1- [E- (3-nitro-2-pyridyldithio) ethyl] -3- (4-methylphenyl) triazene. A process for the preparation of a compound according to claim 1, characterized in that at least one equivalent of a thiol of the formula R3Alk1SH or the formula R4SH- is reacted with one equivalent of a mitosane of the formula Ib 30 35 ß? Wherein R 1, R 3, R 4, Alk 1 and Alk 2 have the meanings given in claim 1, and optionally in the presence of at least one equivalent of a base in an inert solvent at a temperature. temperature from 00 to 60 ° C until a substantial amount of the product of formula II or formula III has been obtained. 18. A process according to claim 17, characterized in that Alk 2 is ethylene and R 1 is hydrogen. IN 19. A process according to claim 17 or {8, characterized in that an equivalent base is used in the reaction. Process according to Claim 17 or 18, characterized in that a lower alkanol, a lower alkanoic acid (lower alkyl) ester, a lower aliphatic ketone, a cyclic aliphatic ether or a lower polyhalogenated aliphatic hydrocarbon having up to 8 carbon atoms or water.