JPS61112078A - Substituted 7-oxomitosane - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION BACKGROUND OF THE INVENTION Field of the Invention The present invention provides novel mitomycin analogs (class 260, subclass 326.24) containing disulfide groups and methods for their preparation. These compounds are mitomycin Al analogs having an organic substituent in which a 7-alkoxy group is combined with a disulfide group.

The present invention relates to mitomycin A and its derivatives (class 2
60, subclass 326.24). Mitomycin A is an antibiotic with established efficacy, and its 7-0-substituted mitosan! (The nine bodies have similar effects.

Nomenclature Systematic chemical abstract name for mitomycin A based on recent revisions [Shirhat
a) et al., Journal of the American Chemical Society (J, Am, Chem, Soc,), 105
.

7199 (1983)) is: CLa5-(laβ, 8β,'i3aα, 3bβ)]-
8-(((aminocarbonyl)oxy)methyl)-6,
8a-dimethoxy-1,la,2°8.8a,8b-hexahydro-5-methyl-azilinoC2',3',3.
4) Pyrrolo[1°2-a]indole-4,7-sion, according to which the azilinoviroloindole ring system is numbered as follows: Chemical Abstracts Cumitomycin Extensive use in the literature The accepted conventional system nomenclature identifies the ring system containing some of the characteristic substituents of mitomycin as mitosane.

Mitosan According to this system, mitomycin A is 7,9a-dimethoxymitosan, and mitomycin C is 7-amino-
9a-methoxymitosan.

Regarding the stereochemical configuration of the products of the invention, when identifying them by the basic name "mitosan" or by the structural formula, it is intended to identify the stereochemical configuration as similar to that of mitomycin A or C.

Prior Art Mitomycin C is an antibiotic produced by fermentation, currently under Food and Drug Administration approval for the treatment of disseminated adenocarcinoma of the stomach or pancreas, in proven combination with other approved chemotherapeutic agents. Mutamycin is sold as a palliative treatment when other physical therapies are ineffective.
ycin) (registered trademark) Bristol Laboratories, 5yra
Cuse, New York 13221), Physicians'Desk Reference
Reference) 37th edition, 1983, pp74
7 and 748). Mitomycin C and its production by fermentation was submitted to Japan on April 6, 1957.
[19] Claiming priority from an earlier application including the application
U.S. Patent No. 3,660.5, issued May 2, 1972.
This is the title of No. 78.

The structures of mitomycin A, BXC and porphyromycin were originally developed by Lederle Laboratories Division American Cyanamid Company (LederleLabo).
Ratories Division America
n Cyanamid Company) web (J
, S. Webb) et al., Journal of American
Chemical Society (J, Am.

Chem, Soc,), 84.3185-3187 (
1962). One of the chemical transformations used in this structural study for mitomycin A and mitomycin C was the formation of the former by reaction with ammonia, 7.
9a-dimethoxymitosan, the latter 7-amino-9a
-methoxymitosan. Substitution of the 7-methoxy group of mitomycin A has proven to be a rather interesting reaction in the preparation of antitumor active derivatives of mitomycin C.

Recently, it was shown that the stereochemical configuration of positions 1.1a, 8a and 8b is as indicated previously with respect to Chemical Abstracts Nomenclature [Silata (Sl+1rh
ata) et al., Journal of the American Chemical Society (J, Am, Chem, Soc,),
105.7199-7200 (1983)). Early literature indicates enantiomers.

The following publications and patents deal inter alia with the conversion of mitomycin A into 7-substituted aminomitomycin C derivatives with antitumor activity. The aim of this work was to produce derivatives that are more active and, in particular, less toxic than mitomycin C. : Matsui et al., Journal of Antibiotics (J, Antibiotics)
, XX [, 189-198 (1968), Konishita et al., Journal of Medicinal Chemistry (J, Med, Chem,), 14.103-1
09 (1971), Iyengar et al., Journal of
Medicinal Chemistry (J, I'led, CI+
Em, ), 14.975-981 (1981), Iyengar et al.
and Bradner), Lecture Proceedings (183rd
Annual Meeting of the American
Chemical Society)
f Papers 183 rd8 natural Me
eting of the America
an Chemical Society, ),
Las Hegas, Nevada, March 1982, Af'tract No. MED I 721)
Sulich et al., U.S. Pat. No. 3,332,944;
Matsui et al., U.S. Pat. No. 3,420,846, 1969, issued July 25, 1967.
Published January 7, Matsui et al., U.S. Pat. No. 3,450,705, 1969.
Matsui et al., U.S. Pat. No. 3,514,452, published January 17, 1970.
Nakano et al., U.S. Pat. No. 4,231.936, published May 26, 1980.
Remers, U.S. Pat. No. 4,268, issued November 4, 2013.
No. 676, published May 19, 1981. The following patent application deals with the preparation of 7-substituted aminomitomycins C, in which the substituents contain a disulfide bond.

Kono et al., European Patent Application No. 116,208 (1984
), Vyas et al., UK Patent Application No. 2,140,7
No. 99 (1984).

7-Alkoxy-substituted mitosans, which are structurally related to mitomycin A, have been described in the paper by Urakawa et al.
of Antibiotics (J.

^ntibiotics), ■, 804-809 (1
980) as a useful antibiotic with activity against tumors in experimental animals.

Mitomycin C is the main mitomycin produced by fermentation and is the commercially available form. Current methods of converting mitomycin C to mitomycin A have many deficiencies. 7-hydroxy-9a corresponding to mitomycin C
Upon hydrolysis to -methoxy-mitosane, diazomethane is then required for the methylation of the material, which is a very difficult material to handle on a manufacturing scale, and the 7-hydroxy intermediate is very unstable [ Matsui et al., Journal of Antibiotics (J.

Antibiotics), XXI, 189-198 (
1968)).

One attempt to circumvent these difficulties involves the use of 7-aziloxymitosans (Kyowa Hakko Kogyo KK'
EE Patent J56073-085, Farm Dock (F
armdoc) No. 56227D/31), Urakawa et al., Journal of Antibiotics (J, Antibiotics), 23.804~
Mitomycin A described by 809 (1980)
Alcoholysis is limited to the preparation of only specific 7-alkoxy structure types due to the availability and reactivity of the alcohol starting materials.

SUMMARY OF THE INVENTION The present invention relates to a group of mitomycin A analogs having a dithioorganic substituent combined in an alkoxy group in the 7-position.

These compounds can be represented by the following general formula: where R2 is an organic group, ie a constituent of the organic thiool of formula R25H, and A I K z and R- have the meanings given below. These compounds are alternatively described by the formulas (1) and (2) or their non-toxic formulation-acceptable salts.

In the formula, A I K r is R3 through its carbon atom
K has 1 to 6 carbon atoms when bonded to R
3 through its sulfur, oxygen or nitrogen atom! K
is a straight or branched alkylene group having 2 to 6 carbon atoms (in which case R3 and -SS- are bonded to different carbon atoms), and 2 to Aβ is bonded to it. With the sulfur and oxygen atoms /l! and any optional A substituents bonded to through oxygen, sulfur or nitrogen are bonded to /lKz different carbon atoms, said substituents being 1 or 2 C3_6 alkyl, C+~, alkanoyl , C1-, alkoxy, halogen, CI-6 alkoxycarbonyl, cyano, C4-6 alkylamino, Cl-6 dialkylamino, CI-6 alkanoylamino and CI-6 alkoxycarbonyl, is a straight or branched alkylene group having from 2 to 2 carbon atoms, AlK and AlK2 can contain a double bond, and R1 is hydrogen, lower alkyl, lower alkanoyl, benzoyl or , substituted benzoyl in which the substituents are lower alkyl, lower alkoxy, halogen, amino or nitro, R3 is halogen, carboxy, alkanoyloxy having 1 to 7 carbon atoms, oxygen atom is 3 to 6 carbon atoms hydroxy attached to ANKI with 1 to 12 carbon atoms, alkylamino or dialkylamino with 1 to 12 carbon atoms, N-alkoxy-alkylamino with 2 to 7 carbon atoms, alkanoylamino with 1 to 7 carbon atoms , to henshylamino or B-substituted hendhylamino [naphthoylamino or B-substituted naphthoylamino, phenylamino or B-substituted phenylamino, cycloalkyl each having 3 to 8 ring members or B-substituted cycloalkyl, each 5 to 8 ring members] cycloalkenyl or B-substituted cycloalkenyl, phenyl or B-substituted phenyl, naphthyl or B-substituted naphthyl, having 1 to 2 rings with 3 to 8 ring members in each ring and selected from oxygen, nitrogen and sulfur in each ring heteroaromatic and heteroalicyclic groups having 1 to 2 heteroatoms; pyridylamino or thiazolyl amino; alkoxy or argylthio each having 1 to 6 carbon atoms; alkoxycarbonyl or alkylaminocarbonyl with 7 carbon atoms, aminocarbonyl, phenoxycarbonyl or B-substituted phenoxycarbonyl, phenoxy or B-substituted phenoxy, naphthoxy or B-substituted naphthoxy, alkoxycarbonylamino with 2 to 6 carbon atoms, Ureid (NHCON
Hz), N-alkylurein with 2 to 7 carbon atoms (-NHCONII alkyl), N3-haloalkylurein with 3 to 7 carbon atoms, N3-haloalkyl with 3 to 7 carbon atoms -N:I-nitrosourein, dialkylaminocarbonyl having 3 to 13 carbon atoms, dialkylaminoalkoxy having 4 to 13 carbon atoms, alkanoylaminoalkoxy having 3 to 7 carbon atoms, and each 2 Hydroxyalkylamino or N, N- with ~8 carbon atoms
dihydroxyalkylamino, said device fI4 group being selected from the group consisting of one or two lower alkyl, lower alkanoyl, lower alkoxy, halogen, amino, carboxy, hydroxy and nitro groups, and R4 is 1 ~alkyl having 12 carbon atoms, each 3 ~
alkenyl or alkynyl having 12 carbon atoms, cycloalkyl or B-substituted cycloalkyl having 3 to 8 ring members, cycloalkenyl or B-substituted cycloalkenyl each having 5 to 8 ring members, phenyl or B-substituted phenyl, naphthyl or B-substituted naphthyl, heteroaromatic and heteroalicyclic rings having 1 to 2 rings, 3 to 8 ring members in each ring, and 1 to 2 heteroatoms selected from oxygen, nitrogen and sulfur in each ring A heterocyclic group is selected from the group consisting of a heterocyclic group selected from the group consisting of a heterocyclic group selected from the group consisting of a
That is, the carbon atom bonded to -8S- cannot itself be bonded to two other heteroatoms), the B substituent may contain 1 to 2 lower alkyl, lower alkanoyl, lower alkoxy, halogen, amino, selected from the group consisting of carboxy, hydroxy or nitro groups, R4 and the adjacent sulfur atom together constitute S-cysteinyl, which S-cysteinyl group is esterified and salt-formed within the non-toxic and non-allergenic peptide. or can be combined.

The compounds of the invention are inhibitors of experimental tumors in animals. In particular, the substances identified as compounds of Examples 17.20 and 21-34 are novel substances. They are used in a similar manner to mitomycin C. The dosages used are adjusted by the ratio of their toxicity compared to that of mitomycin C.

Higher doses are used if the new compound is less toxic.

In another aspect of the present invention, novel methods for producing mitosans of formulas (1) and (2) are provided. This new method includes formula ■, mitosan of formula ■ or formula VI, Ar-N "" N-NH-Allh-SS-AIK+
-R3VAr-N = N-NH-AIKz-SS-R
'Vl (in the formula, R', R3, R4, Al
K1 and AfKz are as described above, and Ar is an organic residue of a diazotizable aromatic amine) with triazene.

Other methods of preparing mitosan of formulas (1) and (2) are provided in variations of the invention. This method requires the formula ■ or ■
, R3AIK+SHR'SH ■■ thiol and formula Ib.

(b) Reaction with mitosan derivatives is included. Disulfide mitosane No. 21b is produced by the triazene method described above. More particularly, mitosan of formula [b, where AffiK2 is ethylene and R1 is hydrogen, is described in Example 20 and Co-pending Application No. 646,888, filed September 4, 1984.

In another aspect of the present invention, the formula (1) (wherein R5 is hydrogen or 01-6 alkyl and R6 is 01
~1□alkyl or substituted C3-I2 alkyl, C1-
I2 cycloalkyl or m c , CI~6 alkanoyl, C6-14 aroyl, cyano, trihalomethyl,
Amino, cI~.

Monoalkylamino, 02-,2dialkylamino, C
b~1□aryl, C6~l□aryloxy, c1~
h alkanoyloxy, 07-,470yloxy, 1
alkoxy, alkanoyl, aroyl, aryl, aryloxy, Alkanoyloxy, aroyloxy, and heterocyclo substituents are each optionally halogen, C1-h alkoxy, Ct-6 alkanoyl, cyano, trihalomethyl, amino, 01-. Alkylamino or C! An improved method is provided for producing a compound having 1 to 2 substituents selected from dialkylamino groups.

The compound of formula (1) is a known compound that has inhibitory activity against many in vivo experimental animal tumors. A number of new compounds corresponding to formula (1) were also prepared by this method and are considered as part of this invention. In particular, the substances identified as compounds of Examples 14, 15, 16 and 19 are novel substances and also have antitumor activity against experimental animal tumors. These compounds are part of this invention. They are mitomycin C
used in a similar way. The dosages used are adjusted proportionally to their toxicity relative to that of mitomycin C. Higher doses are used if the new compound has less toxicity.

A novel method for preparing compounds of formula (1) includes the preparation of mitosane of formula (which is an organic residue of a group amine) with a triazene.

"Lower alkyl" used in the specification and claims
, "lower alkoxy" and "lower alkanoyl" mean (unless otherwise indicated) straight-chain or branched alkyl, alkoxy or alkanoyl groups containing from 1 to 6 carbon atoms, such as methyl, ethyl, Means propyl, isopropyl, butyl, isobutyl, [-butyl, amyl, hexyl, etc. Preferably these groups contain 1 to 4 carbon atoms, most preferably they contain 1
or 2 carbon atoms. Unless otherwise indicated in the individual case, "
The term ``halogen'' shall include chlorine, fluorine, bromine and iodine. The term "non-toxic formulation-acceptable salts" is intended to include the salts of compounds of Formulas I and 1 with non-toxic formulation-acceptable acids or bases. Such acids are well known and include hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, maleic acid, fumaric acid, succinic acid, oxalic acid, benzoic acid, methanesulfonic acid, tartaric acid,
These include citric acid, camphorsulfonic acid, and levulinic acid. Such bases are well known and include, for example, non-toxic metal salts such as sodium, potassium, calcium and magnesium, ammonium salts and non-toxic amines such as trialkylamines, protylenes, dibenzylamines, pyridine, N-methylmorpholine. , N-methylpiperidine, and the like. Salts are manufactured by methods known in the art.

DESCRIPTION OF THE INVENTION The present invention provides a new process for preparing compounds of formula (1) comprising the reaction of mitosane of formula

Scheme 1 where R5 and R6 are as described above and Ar is an organic residue of a diazotizable aromatic amine. 1-Alkyl-3-aryl l-IJ acenes constitute a class of reagents known to be useful for reacting with carboxylic acids to form the corresponding lower alkyl esters. -Methyl-3-(4-methylphenyl)triazene was synthesized by organic synthesis (Org, Syn, ) by White (E, H, White) et al.
48.102-105 (196B) and can be prepared by the general procedure described in Procedure 1. However, this procedure only works well with water-soluble amines, and White (E, H, White) &
rs), No. 21.761 (1961) and herein described in Procedure 2, is more suitable for the preparation of water-insoluble amine triazenes.

The reagent l-methyl-3-(4-methylphenyl) triazene prepared as described above is conventionally used as a carboxylic acid, e.g.
．． The methyl ester of 4-dinitrobenzoic acid [E, H, White, Organi. Cephalosporanic acid (Mangia), Tetrahedron L, which occurs without isomerization
etters), No. 52-, pp. 52
19-20 (197B)). The reagent was also used to prepare 3-methoxy-cef-10 sporin derivatives by reaction with the corresponding 3-hydroxy-3-cephem-4-carboxylade in benzene solution at reflux temperature [Biederkehr ( Wiede
Rkeher et al., U.S. Pat. No. 4,069,324;
Published January 17, 1978].

Other 1-(lower alkyl)-3-aryl triazenes of formula >1 can be similarly prepared by reaction of other lower alkyl amines with aryldiazonium salts. Easily forms diazonium salt 6~
All amines with 12 carbon atoms have 1
, 3-disubstituted triazenes can be used as a source of the aryl moiety. Some examples of triazenes produced by this method and used in the present invention are: 1-(n-butyl)-3-(4-methylphenyl)triazene, 1-(1-methylethyl')-3- (4-methylphenyl)triazene, ■-(4-methylphenyl)-3-(2-(4-morpholinyl)ethyl)triazene, ■-(4-methylphenyl)-3-(2-(2-pyridyl)ethyl ] Triazene, ■-(2-benzylthiolethyl)-3-(4methylphenyl)triazene, 1-(4-chlorophenyl)-3-(2-methoxyethyl)triazene, ■-(4-chlorophenyl)-3-( 1,3-dioxol-2-ylmethyl)triazene, 1-(4-chlorophenyl)-3-(tetrahydrofuran-2-ylmethyl)triazene.

Other triazenes that are suitable reactants for use in the present process to provide 7-(substituted alkoximitosans) of formula (1) have been described in the literature. Daniels (T, A, Daniel
s) et al., Canadian Journal of Chemistry (Can, J. Cam,), 55.3751-37.
54 (1977) is typical.

aX=H, Y=CN bX=NOz, Y=CNcX-CO
zMe, Y=CNdX=Ac, Y=CNeX=NOx, Y=COzEtf
X = COzMe, Y = COZE tg
= COzMe , Y = COPhh X =
NOz , Y=CH' (OCH3h) We now further illustrate suitable triazene starting materials of formula XI for use in the present invention.

1-(n-butyl)-3-(α-naphthyl)triazene, ■-″”(n-hexyl)-3-phenyltriazene, 1-ethyl-3-(2,4-dimethylphenyl)triazene, 1 -(1-methylethyl)-3-(4-methoxydiphenyl)triazene.

In the production of mitomycin A, 3-methyl-1-(4-methylphenyl)triazene is preferably used as a methylating agent. Preferably at least a 2 molar ratio of the latter per molar ratio of 7-hydroxy-9a-methoxymitosane is used, and the reaction is preferably carried out in a liquid organic solvent relative to the 7-hydroxy-9a-methoxymitosane starting material. Preferred solvents are lower alkanols, lower alkyl esters of lower alkanoic acids, di-lower alkyl ethers, cycloaliphatic ethers and lower borino\logenated aliphatic hydrocarbons. These solvents contain up to 6 carbon atoms, but preferably those which boil at temperatures below 100°C. Particularly preferred solvents are methylene chloride, methanol, diethyl ether, ethyl acetate and mixtures thereof.

The reaction can be carried out at the reflux temperature of the reaction mixture or up to about 60°C. At temperatures above this, the mitosan reactant tends to decompose, resulting in reduced yields. Preferably, the reaction is carried out at room temperature or below, for example in the range from 0 to 25°C.

A convenient method of determining when the reaction is complete is thin layer chromatography. Mitomycin A is deep purple in color and can be easily distinguished from the starting material and by-products.

Mitomycin A in the solvent system methylene chloride/methanol (90/10) exhibits Rf=0.36. Chromatography on neutral alumina can be used to purify the product.

The reaction conditions and precautions described above are generally applicable to the preparation of other 7-R60-mitosans by this method.

The novel process of the present invention using 1-substituted-3-aryltriazenes can also be used for the preparation of compounds of formula 1 or 2, which can be obtained by combining mitosane of formula 2 with mitosane of formula 1 or reaction with triazene.

Scheme 2 %Formula% (wherein R', R3, R', AIKI and AIK
Aryltriazenes of the formula (I is as described above and Ar is an organic residue of a diazotizable aromatic amine) have the alkyl amine of the formula xn, R"-8S-AIKZNH2X XI and XIV can be prepared in the same manner as described above for the preparation of aryl triazenes of formula XI, except for substitution with the amino disulfide of, as described by XI and XIV.

Amino disulfides of formula x■ and formula XIV are known compounds and can be produced by various methods. For example, a suitable thiol R'A I K, SR or R'
SH and formula, N HzA I KzS S ()+Na X
The Bunte salt of V or the formula NH2AIKZSSCOCHI XVT can be prepared by reaction with sulfenylthiocarbonate.

Klayman et al., Journal of
Ofgani・7ri・Chemistry (J, Org, Chem
), 1E, 3737-3738 (1964) prepared the following 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 was found to be the preferred reaction solvent for the reaction of Bunte salt and thiol. A reaction temperature of θ to -10°C was found to be preferred for the use of this solvent. Higher temperatures were required with other solvents.

The main drawback of this method is that symmetrical disulfides are formed as by-products, possibly as a result of the disproportionation of the desired hybrid disulfides.

The mixed disulfide starting materials of formulas X■ and XIX are preferably prepared by reaction of a suitable thiol with a sulfenylthiocarbonate of formula XVI.

This has been published by Brois (S., J.) et al., Journal of the American Chemical Society (J.

Am, Chen+, Soc, ), 92.7629~7
631 (1970). Typically, this preliminary procedure involves adding the thiol to a methanolic solution of the amino-alkylsulfenylthiocarbonate of formula XVI and allowing the reaction to proceed at a temperature within the range of 0-25<0>C. Reaction times vary from virtually instantaneous to several hours depending on the particular thiol used. Progress of the reaction can be followed 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 promoter.

1-(Substituted disulfide)-3-aryl triazene of formula (1) or (2) is produced by reacting the amino disulfide of 2. be done. Arylamines having 6 to 12 carbon atoms that readily form diazonium salts can be used as a source of the minor moiety of 1,3-disubstituted triazenes. Some examples of disulfide triazenes made by this method and used in the present invention are L-(2-(2-acetamidoethyldithio)ethyl)
-3-(4-methylphenyl)triazene, 1-(2-(3-nitro-2-pyridylthio)ethyl)-3-(4-methylphenyl)triazene.

Suitable triazene starting materials of formula 1 or formula 2 for use in the present invention are further exemplified below: 1-[2-(3-nitro-2-pyridyldithio)ethyl]-3-(4-chlorophenyl)triazene; 1-[2-'(3-nitro-2-pyridyldithio)propyl]-3-(4-methylphenyl)triazene, 1-[2-(2pyridyldithio)ethyl l-3-(L
t-methylphenyl)triazene, V=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 l -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 invention, formula 1a-.

[wherein R2 is a backing group, i.e. a constituent of the organic thiol of the formula R''SH, which may alternatively be R'AI!k
l or R' (wherein R3, R4 and AItkl
is as described above)] is provided.

For the production of disulfide mitosan of formula 1a, 9 of formula X■
Preferably, a-methoxy-7-[2-(3-nitro-2-pyridyldithio)ethoxycomitosan is used in a thiol exchange process with a suitable organic thiol of formula R''SH as shown in Reaction Scheme 3. The driving force behind the formation of the disulfide of formula 1a is the stability of the by-product, i.e. 3-nitro-2-mercaptopyridine, which modestly
■Exist as.

a ■ X■ + R”SH a When used in the procedure shown, 2lb (wherein /l! and R1 are as above)
A disulfide mitosan having the following is obtained.

Two general synthetic procedures are described herein for the preparation of lipophilic and hydrophilic mitosan of formula 1a. General procedure A is a formula!
General procedure B is preferably used for water-soluble disulfides of formula Ta isolated as sodium salts or as zwitterionic forms. Preferably X
At least 1 equivalent of mercaptan R"SH is used per equivalent of mitosan in
This can be carried out in the presence of about 1 equivalent of base per equivalent of . Preferred bases are tertiary amines such as triethylamine, N-methylmorpholine, N-methylpiperidine,
Pyridine, 2,6-lutidine and inorganic bases such as sodium bicarbonate, potassium carbonate, potassium bicarbonate, and the like. Suitable inert solvents for the reaction of the starting materials of formulas Water. Organic solvents containing up to 8 carbon atoms are preferred, but those boiling at temperatures below 100° C. Particularly 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.

Preferably, the reaction is carried out at room temperature or below, for example within the range of 0 to 25<0>C.

The reaction conditions and precautions described above are generally applicable to the preparation of other dino, rufidomitosan of Formulas 1a and [b] by the general procedure shown in Scheme 3.

The following can be converted by reaction with Bunte salt xv or sulfenylthiocarbonate x to produce intermediates of formula X and XrV, which can then be converted to the products of the invention as described. Figure 1 is a representative 1,000-all enumeration of the formula R3A6klSH or R'SH. In case of preferred embodiments.

Representative thiols can be used in reaction with mitosans of formula lb or lb to produce products of the invention. However, the only limitations to the process of the present invention are the use of thiols containing primary alkyl amines that can be passed into the product mixture and the use of heteroaromatic thiols that cannot react with compounds of formula Ia or Ib. is of use.

SCH3 HSCHIC) (3 H S C HzC HtC H3 HSC) ( (CHi)z H S (C)it)zcHs H S C HzC H (C Hz)zS HS-CH, -C)I=C) (Z H S C Hz C H = C (C
H:l) 2HS-CH2-C3C11 HS-CH. -C3C-C H3H S (C
H2), 10 R'n=2-4;R'=H
, CCH3, ClhR3(CHz), CXR n=1-3; X=1101 NH,NR';
R/ R' = H, CH3 H3(CHz)ll NHR'n""2-4;R' x CH,, CH, CH,.

CHzCHtCHs, CCH3H3(CHz)
-NR'R rich n-2-4; R1/R" -CH5, CHzCHsH
S CHt CHt S CHsH3CHzC)I
tNHC(CHs)iR' -CR3, CCH3 R' = CH3; R2=H, CH3Hz lh ■ NHCOCHzcHgcHco□■ ■ HS -CH,C1 C0NHC1hCO! H HS CH= CH-N HCCH3R' =
CR3; R” = H, CH3NHCH.

NH8N(CHi)z n=2-4; R'=OCH2CH3n=1. 2:
X=0. S, NH n=1. 2:X-0,S, NHn=1. 2
;X−0,S, NHn=1+ 2:X”O,
S, NHn=1. 2, X-0,5,NH n=1. 2, X-0,S, NH N(CH3)z X = O, NH, N CH3, S R' = H, CH3 1.8. △7°～N(CHi)z H 118/\/ \/\. □ /\10H The usefulness of compounds of Formulas I and ■ in the anti-tumor therapeutics of the present invention was demonstrated by an in vivo screening procedure in which the compounds were administered at various dosages to mice induced with P-388 leukemia or BI3 melanoma conditions. This is shown by the results.

The compounds according to the invention appear to have antibacterial activity against Gram-positive and Gram-negative microorganisms similar to that observed for naturally occurring mitomycin, and are therefore potentially useful as therapeutic agents in the treatment of bacterial infections in humans and animals.

Activity against P-388 murine leukemia Table I shows tumor inocula of P-388 murine leukemia lO'ltl aqueous cells implanted into the peritoneal tissue and CDF treated with various doses of the test compound of formula f or ■ or mitomycin C. , including results from laboratory tests on mice. Compounds were administered by intraperitoneal injection. Groups of 6 mice were used for each dose and were treated with Compound 1 on the day of inoculation.
Treated with multiple doses. A group of IO saline-treated control mice was included in each series of studies. A mitomycin C treated group was included as a positive control. Using a 30-day protocol, the mean survival time (
days) were determined for each group of mice and the number of survivors at the end of the 30 day period was recorded.

Mice were weighed before treatment and on day 6. Changes in body weight were taken as a measure of drug toxicity. Mice weighing 20 g each were used, and weight loss of up to approximately 2 g was considered not excessive.

Results were determined by %T/C, which was the ratio of the mean survival time of the treated group to the mean survival time of the saline-treated control group multiplied by 100. Saline-treated control animals generally died within 9 days. In the table below, the maximum effect J is shown as %T/C, and the dose that produces that effect is shown. The values in parentheses are the values obtained with mitomycin C, which served as a positive control in the same study. , a measure of the relative activity of this substance towards mitomycin C can be evaluated. The minimum effect in terms of %T/C was considered to be 125. The minimum effective dose shown in the following table is The two values given on each side in the "Average Body Weight Change" column are the mean body weight change per mouse at the maximum and minimum effective doses, respectively.

Activity against BI3 melanoma Table 1 shows the results of an antitumor test using BI3 melanoma grown in mice. BDF and tumor transplanted tissues were subcutaneously inoculated into mice. A 60 day protocol was used.

Groups of IO mice were used for each dose tested, and the mean survival time for each group was determined. Inoculated in the same way as the test animals,
Control animals treated with injection vehicle and no drug showed a mean survival time of 24 days. Survival time compared to control survival time (%T/C) was used as a measure of efficacy to determine the maximum and minimum effective doses for each test compound.

The minimum effective dose was defined as the dose exhibiting a %T/C value of 125. For each dosage level, test animals were treated with the test compound on days 1.5 and 9 by the intravenous route.

Sunny W 20 110 (112) 3.2 (3)
3.2 +0.5; +0.
526 152 (145) 1.6 (3)
<1゜6 -0.6; -0,61, m
g/kg body weight21) Average of 87 days for each maximum and minimum effective dose3;
Values in parentheses are for mitomycin C tested in the same study.

In view of the anti-tumor activity observed in test animal tumors, the invention includes the use of the substances of the invention in the inhibition of mammalian tumors. For this reason, they are administered systemically to tumor-bearing mammals in antitumor effective amounts with substantially no toxicity.

The compounds of the present invention are primarily intended to be used by injection in much the same manner and for some of the same purposes as mitomycin C. More or less higher or lower doses may be used depending on individual tumor susceptibility. They are readily distributed as dry drug compositions containing diluents, buffers, stabilizers, solubilizers and ingredients that contribute to drug properties. These compositions are then formulated extemporaneously with an injectable liquid vehicle immediately prior to use. Suitable injectable fluids include water, isotonic saline, and the like.

DESCRIPTION OF PARTICULAR EMBODIMENTS In the following procedures and examples, all temperatures are given in degrees Celsius and melting points are uncorrected. Proton nuclear magnetic resonance ('
HNMR) spectra were obtained using Varian X in pyridine-d or DzO as shown.
L 100, Joel F X-90Q
or recorded on a Bruker WM 360 spectrometer. When pyridine-d is used as a solvent, the pyridine resonance at δ-8,57 is used as an internal reference, and when D, O is used as a solvent, TSP is used as an internal reference. Chemical shifts are given in δ and Kampling constants are given in Hertz.

A splitting button is indicated as follows: s, - doublet; d, doublet; t, triplet; q, quartet; m, multiplet; bs, broad signal; dd, doublet of doublet; Double line; dt, triple double line; The infrared spectrum is calculated by Beckma
n) Model 4240 spectrometer or Nic
olet) 5DXFT-IR spectrometer and is expressed in inverse centimeters. Ultraviolet (UV) spectra were measured using a Cary Model 290 spectrometer or a Hewlil, JPackard spectrometer.
) 8450 A spectrometer equipped with a multi-diode array detector. Thin layer chromatography (TLC) was performed using a 0.25 m Ana 1 t
ech) performed on Silica Kel CF plates. Flash chromatography is performed by Woelm J.
) Neutrile alumina (DCC grade) or Wallum silica gel (32°-63 μm) and the indicated solvents. All solvent evaporations were performed under reduced pressure and below 40°C.

-Alkyl-3-aryl triazenes constitute a class of reagents known to be useful for reacting with carboxylic acids to form the corresponding lower alkyl esters. ■−
Methyl-3-(4-methylphenyl)triazene can be produced as follows.

Step 1: White (E, H, White) et al., Organisox Synthesis (Org, Syn, AE, 10
2-195 (1968) 1-Methyl-3-tolyltriazene p-)luidine (50.2 g, 0.47 mol) at 20
Place in a 21 flask equipped with a 0mβ addition funnel and an effective stirrer and immerse the flask in an ice-salt bath at approximately -10°. A solution of 46.8 g (0.55 mol) of potassium nitrite in 150 ml of water is placed in a dropping funnel and 250 g of crushed ice are added.
and 140 mj2 of concentrated hydrochloric acid.
)-) Add to Luizin. Potassium nitrite solution until starch-potassium iodide test shows positive (Note 1) 1
Add slowly with continued stirring for ~2 hours and stir the mixture for an additional hour to complete reaction of all toluidine.

A solution of p-toluene diazonium chloride is then brought to a pt of 16.8-7.2 with cold aqueous sodium carbonate at 0°, the solution becomes red-orange in color and a small amount of red material precipitates. Transfer the cold neutral solution to a dropping funnel and slowly add to a vigorously stirred mixture of tsog sodium carbonate, 300 mn of 30-35% aqueous methylamine (Note 2) and 100 g of crushed ice in a 3e flask. The reaction mixture is approximately 45
Keep the addition width approximately -10', which requires 3 minutes (note 3). ? 8
The liquid was extracted with 13 parts of ether. The ether extract was dried over anhydrous sodium sulfate and concentrated on a rotary dryer at room temperature to yield crude l-methyl-3-p-tolyltriacene (Note 4).
65 g are obtained. Place this in a water-cooled Y1 flower pot,
Sublimation of triazene at 50° (1 dragon) gives a yellow crystalline sublimate, melting point 77-80°, 43.3 g (0,2
9 mol, 62%) are obtained (total of 5). The sublimate was recrystallized from hexane to give triacene as white needle-like crystals, melting point 80.
5 to 8165°. More conveniently it is dissolved in a minimum amount of ether and the solution is diluted with hexane 2
Dilution by volume and cooling to 0° give tabular crystals with a somewhat yellow tinge, melting point 79-81°. The yield of pure triazene is 33-37 g (47-53%) (
Note 6).

Note +11 Individual tests with potassium iodide starch paper should be performed 1-2 minutes after stopping the addition of potassium nitrite.

(2) Can be replaced with 40% aqueous methylamine.

(3) The reaction is complete when one drop of the solution no longer exhibits a red color due to the solution of β-naphthol in aqueous sodium carbonate.

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

(5) The sublimate contains trace amounts of 1,3-';-p-to' as shown by thin layer chromatography.
) ) Contains Lt triazene. When recrystallized, pure 1
-Methyl-3-1-tolyltriazene is obtained.

(6) This procedure works well only with water-soluble amines.

Procedure 2 below is more suitable for the production of water-insoluble amine triazenes.

Step 2 White (E, H, White) and others, Tetrahedron Letters (Tetrahedron L)
ettery, ), N1121. p, 761 (19
61).

1-n-butyl-3-p-chlorophenyltriazene
-p-chlorobenzenediazonium hexafluorophosphate (recrystallized from acetone-methanol) (2.87 g, 1
A solution of n-butylamine (0.73 g, 10.0 mmol), powdered sodium carbonate (15 g) and dimethyl Formamide (30 ml) was added slowly to the stirred mixture. The solution of the diazonium salt can be used at room temperature, but it is much better if the diazonium salt solution is prepared in a cold separatory funnel maintained at about -50° and then fed. Pure product is generally obtained.

and stirred until a negative test was obtained for 2-naphthol (usually completed within a few minutes). add ether,
The mixture was filtered and the filtrate was washed thoroughly with water and dried.

(The triazene can be isolated at this point and recrystallized from pentane at low temperature.) Step 3 7-Hydroxy-9a-methoxymitosan mitomycin c (2.2 g, 6.6 mmol) is dissolved in 0, IN methanolic solution. NaOH (5Q%)■40mI! and the reaction mixture was stirred at room temperature for 30 hours. Then the solution was
Adjust the pH to about 4.0 with N-HCJ and add ethyl acetate (4X
500 mJ). The combined ethyl acetate extracts were dried (Na2S04) and concentrated under reduced pressure at about 30-35 °C to give a solid residue, which was dissolved in ether and excess hexane was added to give a purple precipitate. . The precipitate was collected and air-dried to give the title compound as a slightly purple powder (
1.4g, 63%).

', HNMR (pyridine-length 5.δ): 2.05 (s,
3H), 2.14 (bs, LH)
, 2.74 (bs, IH) , 3.13 (d
, IH), 3. ．． 24(s, 3H),
3.56 (d, l H), 4.00 (dd
, LH> , 4.37 (d, IH)
, 5.05 (t, IH) , 5.40 (dd
, IH), 5.90 (bs, 2H).

Step 4 Mitomycin A 7-hydroxy-9a-methoxymitosan 100■ (
0.30 mmol) and 100 μm (0.30 mmol) of 3-methyl-t-tolyltriazene were dissolved in 1 ml of methylene chloride and 9 m6 of diethyl ether. The solution was gently refluxed for 6 hours and then stirred at room temperature for 18 hours. TLC [methylene chloride:methanol (90
:10)) showed the appearance of deep purple spots at Rf=0.36 and traces of impurities at R,=0.41. The reaction mixture was concentrated to dryness and chromatographed on WOLM neutral alumina using methylene chloride and methylene chloride:methanol (30:1) as the eluent. Fractions containing components at Rr"0.36 were combined and concentrated to dryness. Precipitation of the dry residue from methylene chloride and hexane gave the title compound as an amorphous purple fine powder (25■, 24
%), melting point 161°.

Elemental analysis Calculated value (C+6H1vNaO6)”C,54
,96:H,5,44;N,12.02 Measured value:C,5
3,96iH, 5,37;N, 11.99 IR (KBr), pm+ax, csI-': 3400.
3300.2950.1700.1630.1575.
1200.1060°'HNMR (Viridine S, δ): 1.82 (S, 3
H), 2.74 (dd, LH), 3.12 (d,
IH), 3.24 (s, 3H), 3.54 (d
d.

LH), 3.96 (dd, IH), 4.02 (
s.

3H) , 4.22 (d, 1) () , 4.84 (
bs.

2H), 5.02 (t, IH), 5.38 (d
d.

IH).

The yield in Procedure 4 increases to 63% by using methylene chloride as the reaction solvent and room temperature for 24 hours.

Step 5 Solid NazCO* in a 250 mJ 109-bottom flask,
A 35% aqueous solution of amine (same amount as in step 1) and water were added, and the suspension was stirred at -5°C (ice-salt bath). This suspension was added with p-chlorohenzenediazonium hexafluorophosphate [Aldrich Chemical Co.
ch Chemical Co, ) ) ice, water, Na
, a cold suspension in CO3 (approximately pH 1 solution) was added dropwise.

After the addition was complete, the reaction mixture was extracted with diethyl ether. Combine the diethyl ether extracts and backwash with water;
Dry (Na!504) and concentrate.

The yellow solid residue was dissolved in hexane-methylene chloride (1:1).
It was purified by column chromatography on a column on Wallum alumina using as eluent ('HNMR was recorded).

Examples 1 to 10 Triazenes 1 to 7 in Table 1 were prepared according to General Procedure 1 above, illustrating the triazene of Example 1.

Triazenes were purified by column chromatography on Wallum alumina.

Triazenes 8 to 10 in Table 1 were produced according to General Procedure 5 above.

Example 11 A solution of 1-C2-(3-nitro-2-pyridyldithio)ethyl]-3-(4-methylphenyl)triazene 4-methylphenyldiazonium chloride is prepared from p-toluidine as described in Procedure 1. and adjust the pl (6.8 to 7.2) at 0 °C as described in the procedure. A solution containing 21.15 mmol of diazonium salt in 45 mj? 5.34 g (20.0 mmol) of 2-(3-nitro-2-pyridyldithio)ethylamine added to the flask at
It was placed in an addition funnel connected to a 250 mg 309-bottomed flask containing 7 g of sodium carbonate and 150 mL of dioxane. 6 ml of saturated sodium carbonate aqueous solution! and 10 g of ice were added to the flask. The flask was cooled in a water bath and the contents mechanically stirred. The diazonium salt solution was then added dropwise from the addition funnel over a period of 1 hour. Once the addition was complete, the reaction mixture was allowed to warm to room temperature and then diluted with ether 40
Extracted in 3 parts of 0 mm.

Drying and concentration of the extract yielded the desired product, which was purified using an alumina-filled column, 1,000 in diameter and 10 inches long, in hexane:methylene chloride (4:1); hexane:methylene chloride (3:1); 2); Hexane:methylene chloride (L: 4); Purification was carried out by chromatography using methylene chloride containing 1% methanol after filtration and evacuation of the column. Appropriate fraction (T
(identified by LC) and concentrated to give the title compound 26
5g was obtained.

Examples 12-19 Solution of 7-alkoxy-9a-methoxymitosane''''19>+*trta 2 strokes ± 1+ji-111 triazene (2,4 eq.) in CH2Cl2:methanol (4:1) of 7-hydroxy-9a-methoxymitosane (prepared in step 3) in CHtCl
z: Added to solution in methanol (4:1). The reaction mixture was stirred at room temperature and the progress of the reaction was monitored by thin layer chromatography (TI, C) CH2Cl2
10% M e OH). The 7-alkoxy-9a-methoxymitosane product appears as a dark purple spot on TLC. When the reaction is judged to be complete based on TLC, the reaction mixture is chromatographed on walm alumina to yield 7-alkoxy-9a-methoxymitosane as an amorphous solid. The resulting products are identified in Table 1 as Examples 12-19.

Example 20 9a-methoxy-? -[2-(3-nitro-2=pyridylthio)ethoxycomitosan (20) 7-hydroxy-9a-methoxymitosan, 580 μm (1,73 mmol) is placed in a round bottom flask and dissolved in 60 ml of methylene chloride. . Approximately 2.5 g of triazene from Example 11 (5,7
mmol) was added to the solution in the flask and the mixture was stirred at 5° C. for 14 hours and then at room temperature for 8 hours. The progress of the reaction was monitored by silica '1'' L C using methylene chloride:methanol (9:1). The reaction was kept at room temperature for an additional 26 hours and then packed with alumina in a 3 inch wide by 12 inch long tube. The solvents used for development and elution were methylene chloride and 0.5% in methylene chloride.
Methanol, 1.0% methanol in methylene chloride, 1.5% methanol in methylene chloride, 2% methanol in methylene chloride and 4% methanol in methylene chloride 200 ml each. Met. The appropriate fractions were combined and concentrated to give the title compound, 470 ml.

Elemental analysis Calculated value (C2Z HZ 3 N s O*
S 2 ): C, 45,65; H, 4,09; N,
11.82 (corrected for CHt C1z 0.5 mol%): Measured value: C, 45,74; H, 4,14; N
, 11.61°IR (KBr), max, cm-': 3440-
3200.3060.2930.1720.1570.
1510.1395.1335.1210.1055°'HNMR (Pyridine-length 1, δ): 1.81 (s, 3
H), 2.00 (BS, IH), 2.6! (
bs, IH), 2.98 (bs, l II
), 3.08 (s, 3H), 3.20 (m, 2
H), 3.39 (d, IH), 3.83 (dd.

IH), 4.07 (d, IH), 4.59-4.89
(m, 3H), 5.21 (dd, LH), 7.
16 (dd, LH), 8.31 (dd, il+
), 8.71 (dd, IH).

The procedure of Examples 11 and 20 was repeated with 2 to 6 in the alkyl group.
By adapting to other ω-(3-nitro-2-pyridyldithio)alkylamines having 5 carbon atoms, mitosan derivatives of the following formula can be prepared.

n=2-6 R' = H, or C1-, Alkyl Examples 21-34 The 7-alkoxydithio-9a-methoxymitosans, 21-34, of Table V are described below, and the general compounds shown in Table ■ Manufactured by procedure A or B. Mitosan compounds 21-3
The physical data of 4 are shown in Table ■.

Procedure A To a deoxygenated solution of mitosan (~0.1 mmol) from Example 20 in acetone (3-5 mJ) was added triethylamine (~1.1 eq.) while stirring under argon, followed by acetone (1-2 mJ). ) mercaptan”
(1 equivalent ft.) dropwise or in portions. In many reactions, reaction progress is monitored by silica gel thin layer chromatography (10% CH30H in CH2C1Z).

Completion of the reaction is indicated by the disappearance of the spot corresponding to the starting material and the appearance of the product spot. At each point, the reaction mixture was concentrated under reduced pressure (at ~30 °C) and the residue was dissolved in CH
t(1') is chromatographed on a neutral wall alumina force column (%" separated from the pyridylthione by-product retained in the CH2Cl2.
The product eluted using CI+30H was further eluted with CH,C
Purified to 1 mass by flash silica gel chromatography using 5-7% CH30H in I12 as the eluent. The main band corresponding to the product is isolated and characterized as amorphous 7-alkoxydithio-9a-methoxymitosan.

Note (a) If the starting mercaptan is impure, 1
An equivalent amount of thiol is required.

(b) High performance liquid chromatography (HPLC) monitoring where the starting mitosan of Example 20 and the product have very close R2 values on TLC.
μBondapak-CIII column] is used.

Procedure B To a solution of the mitosan of Example 20 (~0.1 mmol) in 2-5% acetone 1 in methanol (10 mj) was added saturated pJ a HC03 aqueous solution b (~6 drops), then methanol c ( Mercaptan (1 eq.) in 1 ml) is added. Progress of the reaction is monitored by TLC (silica gel, CH
, (10% CH,OH in 112). At the end of the reaction, the reaction mixture is diluted with water (15 mf) and concentrated under reduced pressure at about 30'C to about 10 mj+. The resulting solution was subjected to stepwise gradient elution (100%
'H2O-H,.

80% CHjOH). The product eluting as the main red band is collected h11 and concentrated to give 7-alkoxydithio-9a-methoxymitosan as an amorphous solid.

If further purification is required, repeat the above chromatography step.

Note (al) Methylene chloride can also be used, but acetone is preferred.

(b) This base is not used when the mercaptan is L-cysteine.

(c) Water is used if the starting thiol is water soluble.

(dl Elution with water separates the yellow pyridylthione byproduct from the product retained on the column.

Engineering o　　　　　O- 7o Z

Claims (39)

[Claims] (1) Acceptable salts of compounds selected from the group having formula II or formula III or their non-toxic preparations; , AlK_1 is R^3 through its carbon atom
has 1 to 6 carbon atoms when bonded to AlK_1, and when R^3 is bonded to AlK_1 through its sulfur, oxygen or nitrogen atom (in that case R^3 and -SS- are bonded to different carbon atoms) ) a straight-chain or branched alkylene group having 2 to 6 carbon atoms, AlK_2 is a sulfur and oxygen atom bonded to it and any optional A substitutions bonded to AlK_2 through oxygen, sulfur or nitrogen. groups are bonded to different carbon atoms of AlK_2, and the A substituent is one or two C_1_-_6 alkyl, C_1
A selected from the group consisting of __~_6 alkanoyl, C_1_~_6 alkoxy, halogen, C_1_~_6 alkoxycarbonyl, cyano, C_1_~_6 alkylamino, C_1_~_6 dialkylamino, C_1_~_6 alkanoylamino, and C_1_~_b alkoxycarbonyl a straight-chain or branched alkylene group having 2 to 6 carbon atoms optionally with substituents, AlK_1 and AlK_2 can contain double bonds, R^1 is hydrogen, lower alkyl, lower alkanoyl, benzoyl or substituted benzoyl in which the substituent is lower alkyl, lower alkoxy, halogen, amino or nitro, R^3 is halogen, carboxy, alkanoyloxy having 1 to 7 carbon atoms, Hydroxy bonded to AlK_1 with 3 to 6 carbon atoms, 1 to 1
Alkylamino or dialkylamino with 2 carbon atoms, N-alkoxy-alkylamino with 2 to 7 carbon atoms, alkanoylamino with 1 to 7 carbon atoms, benzoylamino or B-substituted benzoylamino, naphtho ylamino or B-substituted naphthoylamino, phenylamino or B-substituted phenylamino, cycloalkyl or B-substituted cycloalkyl each having 3 to 8 ring members, cycloalkenyl or B-substituted cycloalkenyl each having 5 to 8 ring members, phenyl or B-substituted phenyl, naphthyl or B-substituted naphthyl, heteroaryl having 1 to 2 rings, 3 to 8 ring members in each ring, and 1 to 2 heteroatoms selected from oxygen, nitrogen and sulfur in each ring. Heterocyclic group selected from the group consisting of aromatic and heteroalicyclic groups, pyridylamino or thiazolyl amino, each 1 to 1
alkoxy or alkylthio with 6 carbon atoms, alkoxycarbonyl or alkylaminocarbonyl each with 2 to 7 carbon atoms, aminocarbonyl,
phenoxycarbonyl or B-substituted phenoxycarbonyl, phenoxy or B-substituted phenoxy, naphthoxy or B-substituted naphthoxy, alkoxycarbonylamino having 2 to 6 carbon atoms, ureido (-NHCONH
_2), N-alkylureylene with 2 to 7 carbon atoms (-NHCONHalkyl), N^3-haloalkylureylene with 3 to 7 carbon atoms, N with 3 to 7 carbon atoms ^3-Haloalkyl-N^3-nitrosoureylene, dialkylaminocarbonyl with 3 to 13 carbon atoms, dialkylaminoalkoxy with 4 to 13 carbon atoms, alkanoylamino with 3 to 7 carbon atoms alkoxy and hydroxyalkylamino or N, each having 2 to 8 carbon atoms;
N-dihydroxyalkylamino, said B substituent being selected from the group consisting of one or two lower alkyl, lower alkanoyl, lower alkoxy, halogen, amino, carboxy, hydroxy and nitro groups; ^4 is alkyl having 1 to 12 carbon atoms, each 3
alkenyl or alkynyl having ~12 carbon atoms, cycloalkyl or B-substituted cycloalkyl having 3 to 8 ring members, cycloalkenyl or B-substituted cycloalkenyl each having 5 to 8 ring members, phenyl or B-substituted phenyl, naphthyl or B-substituted naphthyl, heteroaromatic and heteroalicyclic rings having 1 to 2 rings, 3 to 8 ring members in each ring, and 1 to 2 heteroatoms selected from oxygen, nitrogen, and sulfur in each ring; from the group consisting of heterocyclic groups, where the heterocyclic group is bonded through a carbon atom that is bonded to at least another carbon atom;
Said B substituent is selected from the group consisting of 1 to 2 lower alkyl, lower alkanoyl, lower alkoxy, halogen, amino, carboxy, hydroxy or nitro groups, R
^4 and the adjacent sulfur atom together constitute S-cysteinyl, and the S-cysteinyl group can be esterified, salted or conjugated within the non-toxic and non-allergenic peptide. (2) The compound according to claim (1), wherein AlK_2 is ethylene and R^1 is hydrogen. (3) A compound according to claim (2), having the formula II in which AlK_1 is ethylene and R^3 is acetylamino. (4) The compound according to claim (2), having the formula II in which AlK_1 is ethylene and R^3 is acetyloxy. (5) AlK_1 is methylene and R^3 is 1,2-
A compound according to claim (2) having formula II which is dihydroxyethyl. (6) A compound according to claim (2) having formula II, wherein AlK_1 is ethylene and R^3 is carboxy or a non-toxic pharmaceutically acceptable salt thereof. (7) A compound according to claim (2) having formula II, wherein AlK_1 is methylene and R^3 is 1-carboxyaminomethyl or a non-toxic pharmaceutically acceptable salt thereof. (8) A compound according to claim (2), having formula II, wherein AlK_1 is methylene and R^3 is 1-methylimidazol-2-yl. (9) A compound according to claim (2), having the formula II in which AlK_1 is ethylene and R^3 is dimethylamino. (10) A compound according to claim (2), which has formula III, where R^4 is phenyl. (11) A compound according to claim (2), having the formula III, wherein R^4 is 4-nitrophenyl. (12) A compound according to claim (2), having formula III, wherein R^4 is 4-methoxyphenyl. (13) A compound according to claim (2), having the formula III, wherein R^4 is 4-aminophenyl. (14) A compound according to claim (2) having formula III, wherein R^4 is 2-carboxyphenyl or a non-toxic pharmaceutically acceptable salt thereof. (15) A compound according to claim (2) having formula III, wherein R^4 is 4-nitro-3-carboxyphenyl or a non-toxic pharmaceutically acceptable salt thereof. (16) having formula III, where R^4 is 4-pyridyl;
A compound according to claim (2). (17) A compound according to claim (2) or a salt thereof acceptable for non-toxic preparations, which has the following formula: ▲There are mathematical formulas, chemical formulas, tables, etc.▼. (18) Formula I b, ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (In the formula, R^1 and AlK_2 are defined in claim No.
As defined in paragraph 1)). (19) formula, ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ The compound according to claim (2), which has the following. (20) Claim No. (1) applies to mammals with tumors.
A method of inhibiting the growth of a tumor in a mammal, comprising systemically administering a substantially non-toxic, antitumor effective amount of a compound according to paragraph 1. (21) A pharmaceutical composition comprising a compound according to claim (1) in combination with a formulation-acceptable solvent, diluent, adjuvant or carrier. (22) Formula V or formula VI, Ar-N=N-NH-AlK_2-SS-AlK_1-
R^3 VAr-N=N-NH-AlK_2-SS-R
^4 At least one equivalent of triazene in VI is expressed as formula VI, ▲Mathematical formula, chemical formula, table, etc.▼ IV (wherein R^1, R^3, R^4, AlK_1 and A
lK_2 is the same as defined in claim (1), Ar is an organic residue of an aromatic amine that can be diazotized) and 1 equivalent of mitosan in an inert organic solvent under the reaction conditions. A process for preparing a compound according to claim 1, comprising reacting the product of formula II or formula III at a temperature of from about 0 to 60<0>C until an appreciable amount of the product of formula II or formula III is obtained. (23) The triazene of formula VI is 1-[2-(3-nitro-
2-Pyridyldithio)ethyl]-3-(4-methylphenyl)triazene, the method according to claim (22). (24) formula, R^3AlK_1SH or formula R^4SH
At least one equivalent of thiol and the formula I b, ▲ has a mathematical formula, chemical formula, table, etc. ▼ I b (wherein R^1, R^3, R^4, AlK_1 and A
lK_2 is the same as defined in claim (1). ), optionally in the presence of at least 1 equivalent of base, in an inert solvent at a temperature of about 0 to 60° C. to obtain an estimable amount of the product of formula II or formula III. A method for producing the compound according to claim (1), which comprises reacting up to (25) The method according to claim (24), wherein AlK_2 is ethylene and R^1 is hydrogen. (26) The method according to claim (24) or (25), wherein one equivalent of base is used in the reaction. (27) lower alkanols having up to 8 carbon atoms, lower alkyl esters of lower alkanoic acids, lower aliphatic ketones, cycloaliphatic ethers or lower polyhalogenated aliphatic hydrocarbons or water are used as reaction medium;
A method according to claim (24) or (25). (28) Formula IX, ▲There are mathematical formulas, chemical formulas, tables, etc.▼ IX (In the formula, R^5 is hydrogen or C_1_~_6 alkyl, R
^6 is C_1_ to_1_2 alkyl or substituted C_1_
~_1_2 alkyl, C_3_~_1_2 cycloalkyl or substituted C_3_~_1_2 cycloalkyl, in which the carbon atom bonded to the mitosan 7-oxygen atom has 1 to 2 hydrogen atoms, and said substituent is halogen, C_1_ ~_6 alkoxy, C_1_-_6 alkanoyl, C_6_-_1_4 aroyl, cyano, trihalomethyl, amino, C_1_-_6 monoalkylamino,
C_2_～_1_2 dialkylamino, C_6_～_1
_2 aryl, C_6_~_1_2 aryloxy, C
_1_-_6 alkanoyloxy, C_7_-_1_4
aroyloxy, heterocyclo having 1 to 2 rings and having 5 to 12 ring atoms containing up to 4 heteroatoms selected from nitrogen, oxygen and sulfur; alkoxy, alkanoyl, aroyl, aryl; Aryloxy, alkanoyloxy, aroyloxy, and heterocyclo substituents each optionally include halogen, C1_-_6 alkoxy, C_1_-_6 alkanoyl, cyano, trihalomethyl, amino, C_1_-_6
A method for producing a compound having the formula Formula X I, Ar-N=N-NH-R^6 X I (wherein R^5 and R^6 are as described above, and A
r is an organic residue of a diazotizable aromatic amine) with a triazene. (29) Triazene of formula X I is 3-methyl-1-(4
-methylphenyl)triazene, the method according to claim (28). (30) A process according to claim 27 or 28, wherein at least a 2 molar ratio of triazene with respect to the mitosan of formula X is used. (31) Reaction to mitosan of formula
The method according to item 27) or item (28). (32) Claims in which lower alkanols, lower alkyl alkanoates, di-lower alkyl ethers, lower polyhalogenated aliphatic hydrocarbons or cycloaliphatic ethers having up to 8 carbon atoms are used as reaction medium. The method according to paragraph (27) or paragraph (28). (33) Methylene chloride, methanol, diethyl ether, ethyl acetate or mixtures of two or more thereof are used as the reaction medium,
The method according to item 27) or item (28). (34) The method according to claim (27) or (28), wherein the reaction temperature is 0 to 60°C. (35) The method according to claim (27) or (28), wherein the reaction temperature is 0 to 25°C. (36) R^6 is 2-(benzylthio)ethyl,
The method according to claim (28), wherein R^5 is hydrogen. (37) R^6 is 2-(2-pyridyl)ethyl,
The method according to claim (28), wherein R^5 is hydrogen. (38) The method according to claim (28), wherein R^6 is 2-(4-morpholinyl)ethyl and R^5 is hydrogen. (39) R^6 is (1,3-dioxolan-2-yl)
The method according to claim (28), wherein R^5 is methyl and R^5 is hydrogen.