CA1327037C - Mitomycin derivatives having reduced bone marrow toxicity, processes for their preparation, and the uses thereof - Google Patents

Abstract

ABSTRACT

The invention relates to certain derivatives of mitomycins A and C having good anti-neo-plastic properties and low bone marrow and overall toxicity comprising:

wherein, n is 0 or 1;
Y is selected from the group consisting of glucopyranosyl, galactopyranosyl, mannopyranosyl, xylopyranosyl, cellobiosyl, lactosyl, glucofuranosyl, maltosyl, and 1,3-cyclohexanediol-2-yl or the hydroxyl-protected acetate derivatives thereof;
R is hydrogen;
R1 is hydrogen, C1-C4 alkyl or C1-C4 allyl substituted by phenyl, hydroxyphenyl,indolyl, mercapto, C1-C4 allylthio, hydroxy, carboxy, amino, guanidino, inidazole or carbamyl;
or R and R1 form a five or six membered ring containing nitrogen; or wherein, n, Y, R and R1 are the same as above; and R2 is NH2 or CH3O-; or wherein, R2 is as defined above; and R3 is a 3-cyano-4-morpholinyl-2-deoxypyranosyl saccharide or a 4-morpholinyl-2-deoxypyranosyl saccharide;
and to methods for the preparation and use of the mitomycin derivatives.

Description

-~ ~327~37 IITLE OF THE INVENTION
MITOMYCIN DERIVATIVES HA~IN6 REDUCEP BONE MARRO~ TOXTCITY~
PROCESSES FOR THEIR PREPARAT~ON ~ND THE ~SES THEREOF

FIELD OF THE INYENTION:

The invention is in the field of pharmaceutical agents and the uses thereof.

BACKGROUND OF THE INVENTION

The mitomycins are a family of compounds having the following general formula (I):

.

H3C r~

~ -2- ~327~37 Mitomyc~ns A, B and C are related to one another as set forth in Table 1 below, the designat;ons X, Y and Z being those of formula 1.

MitomYcin: X Y Z

C -~H2 -OCH3 -H
Mitomycins are derived from mitosane compounds having the follow-ing skeleton (II~:

H~C ~

The mitosanes are formed during the cultivation of the micro-organism $treptomvces caesPitosus in a liquid nutrient medium under artificially controlled conditions. After separating the resulting mycellium, the various mitomycins may be isolated from the latter by active carbon or preferably non-ion exchange resin ~dsorption, organ;c solvent extraction or chromatography on alumina, as disclosed in U.S.
Patent No. 3,650,578 to Hata et al~
Although the mitosanes are excellent antibiotics, they have limited utility due to their toxicity to human blood ~see U.S. Patent No. 3,450,705 to Matsui ~t al.). The relatively highly toxin nature of the co~pounds has prompted seareh for derivatives of mitomycin to ~ncrease the antibiotic activity and to decrease toxicity.

A68. 2 .WP 082288 For exa~ple, Matsui et al., U.S. Patent No. 3,450,705, disclose mitomycin compounds substituted at the 7-position with amino, lower alkylamino, phenylamino, or pyridyl, and substituted at the la position with haloalkanoyl, halobenzoyl, nitrobenzoyl, alkenoyl, acetyl glycyl, sorboyl, or acetyl methionyl.
Matsui et al., U.S. Patent No. 3,558,651, disclose mitosane derivatives comprising la-acyl-7-acyloxy-9a-methoxy compounds.
Certain mitomycins and mitomycin derivatives also possess antitumor activity. Oboshi et al., Gann 58:315-321 (1967); Usubuchi et al., Gann 58:307-313 (1967~; Matsui et al., J. Antibiotics XXI:189-198 (1968); Japanese Patent No. 68 06 627 to Matsui et al. (Chemical Abstracts 69:86986k (1968)); and Cheng et al., J. Med. Chem. 20:767-770 (1977)-While mitomycin C is active against a relatively broad spectrumof experimental tumors, its toxicity and myelosuppressive effects limit its use in clinical practice ~MitomYcin 0: Current Status and New Developments, Carter et al. (eds.), Academic Press, New York (1979)). In preclinical and clinical studies, mitomycin C has shown activity against a variety of murine and human neoplasms, but has also shown severe, delayed bone marrow toxicity. Goldin, A., et al., NCI-EORTC Svm wsium on Mitomyc _ C, Brussels, Belgium (1981).
In other studies, a combination of 5-fluorouracil, adriamycin and mitomycin C was found to be effective for the treatment of patients with advanced gastric and colorectal cancer. This regimen incor-porated mitomycin C administration in a single dose schedule every two months, to decrease the treatment-limiting delayed myelosuppressive effects of the compound. Schein, P.S., et al., MitomYcin C Current Status and New DeveloDment~, pp. 133-143, Carter e~ al. (eds.), A~ademic Press, New York (1979~.
Numerous synthetic derivatives of mitomycin C have been prepared in the hope of obtaining compounds with improved therapeutic proper-ties. These derivatives include substitution on the aziridine ring, carbamoyl, or acyl group substituticn on the hydroxymethyl side chain, A68.2.WP 082288 4 1327~37 and replacement of the 7- substituent in the quinone ring with other functional groups, especially substituted amines. However, as disclosed by Remers, U.S. Patent No. 4,~68,676, nnne of these analogs have emerged as a clinical agent, with the possible exception of the 7-hydroxy analog of the mitomycin C, which has been involved in a recent study in Japan. This analog is asserted to be less leukopenic than mitomycin C, but is also less potent. Also disclosed by Remers, suDra, are totally synthetic m1tomycin analogs of the mitnsane type (Mott et al., J. Med. Chem. 21:493 (1978)J, prepared mainly for their antibacterial activity.
Kinoshita, S., et al., J. Med. ChemO 14:103-112 (1971), disclose several derivatives of mitomycin substituted in the la, 7, and 9a positions. In particular, compounds substituted at the la position with sulfonyl, ortho-substituted benzoyl, and acyl derivatives were reported.
Iyengar, B.S., et al., J. Med. Chem. 24:97~-981 (1981), disclose a series of 31 mitomycin C and porfiromycin analogues with various substituents at the 7- and 1a-positions. The most active substituents at the 7-position included aziridine, 2-methylaziridine, propargyl-amine, furfurylamine, methyl glycinate and 3-aminopyridine.
Iyengar, B., et al., J. Med. Chem. 26:16-20 (1983), disclose a series of 7-substituted mitomycin C and porfiromycin derivatives and the screening the~eof in standard antitumor systems. The authors report that the 7- position controls the reduction of the quinone ring, thus suggesting that it would be possible to alter the substitu-tion of the 7- position to gain selectivity between normal cells and certain cancer c~lls.
Iyengar, B.S., et al., J. Med. Chem. 26:1453-1457 (1g83), disclose 20 mitomycin C analogues substltuted with secondary amines at the 7-position. Eleven of these analogues were more aetive than m1tomycin C against P388 murine leukemia and two of these cleven were significantly less leukopenic. The authors report that no quantita-tive correlation between antitumor activity and physiochemical A68.2.WP 08?288 ~ ~327~3~
s-properties of the analogues was e~ident, although the relative ease of quinone reduction may be related to activity.
Iyengar, B.S., et al., J._Med. Chem. 29.1864-1868 (1986), disclose the preparation of 7-substituted a~ino 1,2-aziridinomito-senes.. The authors reported that a ~e~hyl group on the aziridine nitrogen gave increased potency. The 7-amino mitosene derivatives which were difficult to reduce to hydroquinones ~ere essentially inactive.
. Sami, S., et al., J~ Med. Chem. ?7:701-708 (1984)9 disclose a series of 30 N7-phenyl-subst~tuted mitomycin C analogs. Two of the compounds having pyrazolyl or aminopyridyl substituents at the 7-position were disclosed as clearly superior to ~itomycin C in activity against P388 murine leukemia~
Sami, T., et al., J. Med. Chem. ~:247-250 11979), also disclose N-(2-chloroethyl)-N-nitrosocarbamoyl derivatives of glycosylamines, including three disaccharide derivatives which exhibited strong antitumor actiYity against leukemia 1210 in ~ice. In addition, glucopyranos~ derivatives of N-nitrosoureas possess i~munogenic and marrow-sparing properties. Anderson et al., Cancer Research 35:761-765 (1975); Panasci et al., ~L_s~ln,_lcy~s~ 64:1103-1111 (1979).
In U.S. Patent No. 4,720,543, compounds ha~ing the following general formula (III~ are disclosed:

1l ~ff2 ON~2 ~l_f ~CH3 111 H~C~--N~ CS-~tlR 2 A68. 2 .IJP 082288 - 6 - ~L32~37 where R1 is selected from the group consisting of NH2, C1-C4 alkoxy and a glycosyl residue; and R2 is selected from hydrogen, C1-C4 alky], and a glycosyl residue, with the proviso that either R
or Rz, but not both, contain a glycosyl group.
The compounds represented by formul.a III have excellent antineoplastic activity and at the same time possess reduced bone marrow toxicity and lower overall toxicity.
Despite the above-listed mitomycin derivatives, a need continues .: to exist for improved mitomycin derivatives having good anti-neoplastic properties and low bone marrow and overall toxicity.

SUMMARY OF THE INVENTION
The invention relates to a mitomycin derivative having the following general formula (IV):

o o CH20-~-NH2 H

wherein n is 0 or l;
Y is selected from the group consisting of glucopyranosyl, galactopyranosyl, mannopyranosyl, xylopvranosyl, cellobiosyl, .; lactosyl, glucofuranosyl, maltosyl, and l,3-cyclohexanediol-2-yl, or a hydroxyl-protected peracetate derivative thereof;
.

D`

~ 1327~

R is hydrogen;
Rl is hydrogen, Cl-C4 alkyl or Cl-C4 alkyl substituted by phenyl, hydroxyphenyl, indolyl, mercapto, C1-C4 alkylthio, hydroxy, carboxy, amino, guanidino, imidazole or carbamyl; or R and Rl together form a f~ve or six membered nitrogen containing ring.
The invention also relates to a mitomycin derivative having the following general foMmul a (V):

o CH20-~-NH2 R2 ~ ~ C-CH2-CHz-C-(N-CIH;T)n-NH-Y V

wherein n, R, R1 and Y are as defined above and R2 is NH~- or CH30-.
The invention also relates to a mitomycin derivative having the following structural formul a (V I ):

t) CH20-C-~H2 R2 ~N -SCNH- R3 Vl wherein R2 is as defined above; and R3 is a 2-(3-cyano-4-morpholinyl)-2-deoxypyranosyl saccharide or a 2-~4-morpholinylJ-2-deoxypyranosyl saccharide.
The invention also relates t~ a process for preparing a mitomycin derivatiYe havlng the formula (IV) A68.2.~P 9~2288 -~ 32 7~3~

o o ~H2~-C-NH2 Y-NH-(-~-~R~~~)n ~ IV

wherein n is 1;
Y is selected from the group consisting of glucopyranosyl, galactopyranosyl, mannopyranosyl, xylopyranosyl, cello-biosyl, lactosyl, glucofuranosyl, maltosyl, and 1,3-cyclo-hexanediol-2-yl, or a hydroxyl-protected peracetate deriva-tive thereof, R is hydrogen;
R1 is hydrogen, C1-C4 alkyl or C1 C4 alkyl substituted by phenyl, hydroxyphenyl, indolyl, mercapto, C1-C4 alkylthio, hydroxy, carboxy, amino, guanidino, imidazole or carbamyl; or R and ~1 together form a five or six membered nitrogen containing ring;
comprising:
(a) condensing an N-protected amino acid w;th an alcohol in the presence of a dehydration reagent to give an activated ester, (b) condensing the activated ester obtained in step (a) with an amino compound to give a protectcd amino acid amino compound con-jugate, :: ~c) removing the protecting group of the protected amino acid-amino compound conjugate obtained in s~ep (b) to give an amino acid-amino compound con~usate, and : ~ ld~ condensing the amino acid-amino compound conjugate obtained in step (c) with mitomycin A to give the mitomycin derivative.

A68.2.WP 082288 ~27~37 The invention also relates to a process for the preparation of a mitomycin derivative having the formula (VI)o o 1 CH2o_~_NH2 VI
R2 ~ CNH-R3 O
wherein R2 is NH2 or CH30-; and R3 is a 2-(3-cyano-4-morpholinyl)-2-deoxy pyranosyl saccharidç;
comprising (a) condensing bis(acetaldehyde-2-yl) ether with a 2-amino-2-deoxy saccharide in the presence of a salt of cyanoborohydride to give a 2-deoxy-2-(3-cyano-4-morpholinyl) pyranosyl saccharide and a 2-deoxy-4-morpholinyl pyranosyl saccharide;
(b) separation of the 2-deoxy-2-(3-cyano-4-morpholinyl) pyranosyl saccharide from the 2-deoxy-4-morpholinyl pyranosyl saccharide obtained in step (a);
(c) reaction of the 2-deoxy-2-(3-cyano-4-morpholinyl) . pyrano yl saccharide obtained in step (b) with an acetyl halide to give a 2-deoxy 1-halo-2-(3-cyano-4-morpholinyl) peracetyl pyranosyl saccharide;
(d) treatment of the 2-deoxy-1-halo-2-(3-cyano-4-morpholinyl) peracetyl pyranosyl saccharide obtained in step (c) with silver thiocyanate to give a pyranosyl saccharide-1-thiocyanate;

11~' - lo - 1 ~ 2 7 ~ ~7 (e) reaction of the pyranosyl saccharide-l-thiocyanate obtained in step (d) with mitomycin C or mitomycin A to give a mitomycin C- or mitomycin A-pyranosyl saccharide peracetate carbothioamide; and (f) hydrolysis of the acetate groups of the mitomycin-C-pyranosyl saccharide peracetate obtained in step (e) to give the mitomycin derivative.
The invention also relates to a process for the preparatian of a mitomycin derivative having the following formula (VI) CH3~N_CNH-R3 Vl wherein R2 is NH2- or CH30-; and R3 is a (4-morpholinyl)-2-deoxy pyranosyl saccharide;
comprising (a~ condensation of bis(acetaldehyde-2-yl) ether with a 2-amino-2-deoxy pyransoyl saccharide in the presence of a salt of cyanoborohydride to give a 2-deoxy-2-(3-cyano-4-morpholinyl) pyranosyl saccharide and a 2-deoxy-2-(4-morpholinyl~ paranosyl saccharide;
(b) separation of said 2-deoxy-2-(4-morpholinyl) pyranosyl saccharide from said 2-deoxy-2-~3-cyano-4-morpholinyl) pyranosyl saccharide obtained in step (a)i (c) reaction of the 2-deoxy-2-(4-morpholinyl pyranosyl saccharide obtained in step (b) with an acetyl halide to give a 2-deoxy-1-halo-2-(4-morpholinyl) peracetyl pyranosyl saccharide;
(d) treatment of the 2-deoxy-1-halo-~-(4-morpholinyl) peracetyl pyranosyl saccharide obtained in step (c) with silver thiocyanate to give a pyranosyl saccharide-l-thiocyanate;
(e) reaction of the pyranosyl saccharide-l-thiocyanate obtained in step (d) with mitomycin A o~ C to give a mitomycin A- or C-pyranosyl saccharide percetate carbothioamide; and (f) hydrolysis of the acetate groups of the mitomycin-C-pyranosyl saccharide peracetate obtained in step (e) to give the mitomycin derivative.
The invention also relates to a process for the preparation of a mitomycin derivative having the following formula (V) CH3 ~ 3 C-tH2-cll2-c-(-~-clH-c)n-NH-y wherein n is 9 or 1;
Y is selected from the group consisting of glucopyranosyl, galactopyranosyl, : ~ mannopyranosyl, xylopyranosyl, cellobiosyl, lactosyl, glucofuranosyl, maltosyl, and 1,3-cyclohexanediol-2-yl;
R is hydrogen;
R1 is hydrogen, C1-C4 alkyl or C1-C4 alkyl substituted by phenyl, hydroxyphenyl, indolyl, mercapto, C1-C4 alkylthio, hydroxy, carboxy, amino, guanidino, imidazole or carbamyl; or R and R1 together form a five or six mem}}ered nitrogen containing ring;

-12- ~327~37 R2 is NH2-;
comprising:
(a) condensation of mitomycin C with succinic anhydride under basic conditions to give mitomycin C-la-succinic acid ester;
(b) condensation of the ~;tomycin C-la-succinic acid ester obtained in step (a~ with a compound of the Formula (VII).

~(N-cH-~)n-N~l-YP VII

wherein R, R1 and n are defined above and YP is a hydroxyl-protected saccharide selected from the group consisting of the hydroxyl-pro-tected derivatives of glucopyranosyl, galactopyranosyl, mannopyrano-syl, xylopyranosyl 9 cellobiosyl, lactosyl, glucofuranosyl, maltosyl, and 1,3-cyclohexamediol-2-yl;
~ c~ removal of the hydroxyl protecting groups to give the mitomycin derivative.

The invention also relates to a process for the preparation of a mitomycin derivat;ve having the following formula (V):
O cH20-C-NH~ p o n CH3 ~ ~ / C-CHz-CH2-C-(-N-CH-C~-NH-Y V
wherein n is 0 or 1;
Y is selected from the group consisting of glucopyranosyl, g al actopyranosyl, mannopyranosyl, xylopyranosyl, cello-biosyl, lactosyl, glucofuranosyl, maltosyl, and 1,3-cyclo-hexanediol-2-yl;
A68.2.WP 082288 _13- 13~7037 R is hydrogen;
R1 is hydrogen, C1-C4 alkyl or C1-C4 alkyl substituted by phenyl, hydroxyphenyl, indolyl, mercapto, C1-C4 alkylthio, hydroxy, carboxy, amino, guanidino, imidazole or carbamyl; or R and Rl together form a five or six membered nitrogen containing ring;
R2 is CH30-;
comprising:
(a) condensation of mitomycin A with succinic anhydride under basic conditions to give mitomycin A-la-succinic acid ester;
(b~ condensat;on of the mitomycin A-la-succinic acid ester obtained in step ~a) with a compound of the Formula (VII) Il P
~(N-cH-c)n-NH-y VII

wherein R, R1 and n are as defined above and YP is a hydroxyl-pro-tected saccharide selected from the group consist;ng of the hydroxyl-protected derivatives of glucopyranosyl, galactopyranosyl, manno-pyranosyl, xylopyranosyl, cellob;osyl, lactosyl, glucofuranosyl, maltosyl, and 1,3-cyclohexanediol-2-yl;
(c) removal of the hydroxyl protecting groups to give the mitomycin derivative.
The invention also relates to pharmaceutical compositions comprising a therapeutically effective amount of the mitomycin derivatives of the invention together with a pharmaceutically accep-table carrier.
The invention also relates to methods for the treatment of bacterial inf2ctions compr;sing admin;stering the pharmaceutical compositions of the invention to an animal.
The invention also relates to methods for the treatment of cancer by suppress~ng growth of ca~cer cells susceptible to growth suppres-A68.2.~P 082288 -14- ~27~37 sion comprising administering the pharmaceutical compositions of the invention to an animal.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

The synthetic preparation of the mitomycin deri~atives of the invention ha~e, as their starting poin~, mi~omycin C. Mitomycin C may be prepared according to the methods generally disclosed in Cheng et al., J. Med. Chem. 2~:767-770 ~1977~. Alternatively, mitomycin C can be obtained from mitomycin A by treatment of mitomycin A with a methanolic-ammonia solution as described by Matsui, M., et al., J.
Antibiotics XXI:189 (1968~.
The mitomycin derivatives of Formula IV, wherein n=O (X), may he obtained by displacement of the methoxy group of mitomycin A (VIII) with the amino group of an amino compound, for example, glucosamine (Y-NH2; (IX)) under basis conditions in a polar organic solvent to give the N7 substituted mitomycin derivative (X) (see Scheme I below).
Amino compounds (Y-NH2) which may be substituted at the 7-position include, but are not limited to glucosamine, galactosam;ne, mannosamine, xylosamine, eellob;osamine, maltosamine and 2-amino-1,3-cyclohexanediol and the hydroxyl-protected peracetate derivatives thereof. Preferably, the saccharide comprising the group "Y" is substituted at the 2-position with the amino group. Polar organic solvents which may be used ln the practice of the invention include methanol, ethanol, propanol, dimethylsulfoxide, and dimethylformamide.
Suitable bases fsr providing the basic conditions of the reaction include alkylamines such as C1-C3 trialkyl amines, diisopropylethyl-amine, 1,8-d~azabicyclo~5.4.0]undec-7-ene ~DBU) and dimethylamino-pyr;dine (DMAP). In general, mitomycin A and the amino derivative are present in a 1:1 molar ratio, although excess amino derivative may be present. Sufficient base is present in the reaction mixture to insure that the reaction remains basic throughout.

A68.2.WP 082288 1327~7 Preferred mitomycin derivatives having Formula X include N7-(2-deoxyglucopyranosyl)mitomycin C, N7-(2-deoxygalactopyranosyl)mitomycin C, N7-(tetraacetyl-2-deoxyglucopyranosyl~mitomycin C, and N7-(tetra-acetyl-2-deoxygalactopyranosyl~mitomycin C.
Scheme I

o CH~-C-NH2 NHz ~H Mitomycin C

1. NaOH/MeOH

2. CH2N2 o ~H2D-C-NH2 CH30 ~ " OCH3 CH ~ N ~ 'NH Mitvmycin A

¦ Y-NH2 IX

o SH~O- C-NH2 Y-NH ~H X

A6B . 2 . ~P 082 288 ~ ~7~37 The amino acid linked mitomycin derivatives of formula IV, wherein n=l, may be prepared (Scheme II) from mitomycin A by condensa-tion of the N-protected amino acid, for example, the N-benzyloxy-carbonyl derivative (XI)~ with an alcohol such as N-hydroxysuc-cinamide, which is capable of generating an activated ester, and a dehydrating reagent to give the activated ester (XII). Dehydrating reagents which may be used ln this process include, but are not limited to dicyclohexylcarbodiimide (DCC) and diethylazodicarboxylate (DEAD) and triphenylphosphine. Treatment of the activated ester (XII) with any of the above-listed amino compounds (IX) gives the protected amino acid-amino compound conjugate ~XIII). Removal of the protecting group, for example, by hydrogenolysis of the N-benzyloxycarbonyl group, gives the free amino derivative (XIV). Compound (XIV) may then be condensed with mitomycin A (VIII) by displacement of -OCH3 as described abnve to give the amino acid linked mitomycin derivative (IV).
Preferred mitomycin derivatives having Formula (IV~, wherein n=l, Rl = H and R ~ H include N7-[1[(2-deoxy-2-glucopyranosyl)amino]
carbonyl]methyl] mitomycin C and N7-[~[(tetraacetyl-2-deoxy-2-glucopy-ranosyl)amino]carbonyl]methyl]mitomycin C.

A68.2.WP 082288 17 132~

Scheme I I

l~aO~~N~ 01~ CC els2~N--I:a-C-O-N~
N~ ~ ~ R R

¦ Y-NH2, IX

Pd/Cg 8a H ,~1 R
XIV l;~J!I Xlll P
cH R cH2o c NH~

3~ H
~1111 Q o ~H~o-~-hH2 ~-NH-(-~ -N-~n~, ~H3~ N~"~H

IY

A68. 2 .WP 08228 -18- ~3~7~

Mitomycin derivatives having Formula (V), wherein ~ is NH2 and n is O ~Formula (XV), below), may be prepared (Scheme III) by condensa-tion of mitomycin C. (XVI) with succinic anhydride to give the amide ~XVII) which may then be condensed with the hydroxyl-protected amino derivative YP-NH2 (XVIII) using any of the above-listed dehydrating reagents followed by deprotection to give (XV). Protec~ing groups for the amino derivative include, but are not limited to, C2-C4 acyl esters.
Preferred mitomyein derivatives hav~ng Formula (XV) include N1-[[2-[[(2-deoxy-2-glucopyranosyl)amino~carbonyl]ethyl]carbonyl]
mitomycin C.

S~heme IIl Il O O
O CH O-C-NH

CH3 ~ CH3 ~ C-tHZ-CH2 C-~H
XVI XVlI

/1. )P-N~ XV I I I
2. Depro ec~

CH O~ H

~v A68. 2 .WP 082~88 ~7~

Mitomycin derivatives hav;ng Formula (V), wherein R2 is -OCH3 and n is O ~Fnrmula ~XIX~, below), may be prepared (Scheme IV) by treat-ment of mitomyc~n C (XVI) with sodium methoxide in absolute methanol ~o give mitomycin A (VIII) followed by condensation with succinic anhydride to give the mitomycin A-la-succinic acld ester (XX).
Condensation of the carboxylic ac~d group of (XX) with the hydroxyl-protected amino derivative YP-NH2 (XVIII), as described above, followed by deprotection g~ves (XIX).
Preferred mitomycin derivatives having Formula XVIII include Nl-[[2-[~(2-deoxy-2-~lucopyranosyl)aminoJcarbonyl]ethyl]carbonyl]
mitomycin A.

Scheme IY

o CH20-c-NH2 R
NH2 ~ 1. NaOHIMe~

~Xvl) O ~ cS
o CH~o_C_~H
C~O ¦ ~ ~CH3 -cH2-c-NH-Y O
~3 ~ ~ O CH20-C~NH,2 XIX p ~ ~3~ ,0 ~C~3 ~ ~
1. Y -NH XVI I I \ ~ CH~-cH2 ~-~H
2. Depro~ect ~H3 ~N~; N

XX

A68.2.iP 082288 ,;

~ ~.327~7 -2~-The mitomycin derivatives of Formula YI may be prepared according to the sequence depicted in Scheme Y. Treatment of 3,4-dihydroxy-tetrahydrofuran (XXI) with aqueous sodium periodate in a polar organic solvent gives bis(acetaldehyde-2-yl) ether (XXII) which may be con-densed with a 2-amino-2-deoxy-saccharide (XXIII) in the presence of a salt of cyanoborohydride to give a mixture of 2-deoxy-2-(3-cyano-4-morpholino) saccharide (~XXIVa), ~ - -CN), and 2-deoxy-4-morpholinyl saccharide ((XXIVb), Q ~ -H) which may be separated, for example, by column chromatography. Sal~s of cyanoborohydride may include any of the alkali metal salts of cyanoborohydride, preferably sodium cyanobo-rohydride. Treatment of the saccharide deriva~ive (XXIVa) with an acetyl halide gives the 2-deoxy-1-halo-(4-morpholinyl) peracetyl saccharide which may be reacted with silver thiocyanate to give a 1-thiocyanate saccharide (XXV). Condensation of the thiocyanate (XXV) with mitomycin C (XVI) gives the mitomycin C-saccharide peracetate carbothioamide (XXVI). Deacylation of (XXVI), for example, with methanolic ammonia, gives (VI) (Q = -CN or H).
Preferred mitomycin derivatives having Formula VI include 2-(3-cyano-4-morpholinyl) 2-deoxyglucopyranosyl mitomycin-la-carbothioamide ~nd 2-(3-cyano 4-morpholinyl)-2-deoxygalactopyanosylmitomycin-la-carbothioamide.

A68.2.WP 082288 -21- ~3~71~7 Scheme V

f ~20H C132C~13 1~0 ~ N~194 ~,eB~ O ~ ~ ~ N~ 3e~ ~ y ~C:H2~ 30 BO~OH I~O~OH
XXI XXII N~12~C~
XXIII ~ J
XXIV
1. AcBr 2. AgSC~I

NH2~--N 2 ~O NCS
l ~Q\ ~NHC- Mitomycin C ~?Ac / ~A~ c~ ~
A~ ~, . ~N~ Q
rQ ~ XXV ~oJ
~o~ \ NH3lcH3oH
XXVI
N~32- co-~H2 CH2~H ~33~ 2 NHC-~p~LCH3 ~0~

A68 . 2 . WP 082288 ~3~7~

The compounds of the invention may be present as pharmaceutically acceptable salts. Among the preferred an;onic counter ions are those of the halides (derived from hydrohalic acids~, such as chloride, brom;de, or fluoride. Other anions include sulfonate, or D-toluene-sulfonate.
As an antibiotio, the compounds of the present invention are useful aga;nst all microorgan;sms susceptible to the anti-bacterial action of the parent compounds, these microorganisms including, but not limited to, Pseudomonas, $taDhYlococcus, Sarcinia, D;plococcus, Str~ptococcus, CorYnebacterium t ~emo~hi1us, Escherichia, Klebsiella, Proteus, Salmonella, Shiaella, Brucella, MYcobacterium, Nocardia, SaccharomYces, Cand;da, Penicill;um, and As~erqillusO Specific microorganism treatable with the compounds of the present invention include Pseudomonas aer~q~_osa, Staphvlococcus aureus, StaphYlococcus albus, StaDhvlococcus c treus, Sarcina lutea, Diplococcus Dneumoniae, Streptococcus hemol Yti CU$, strePtococcus lactis, CorYnebacterium diphtheriae, Hemophilus pertussis, Escherichia coli, Klebsiella Pneumoniae, Proteus vulqaris, Salmonella _~yPhosa, Salmonella Para-~Yeh~, Shiqella dYsenteriae, Brucella abortus, Brucella meqatherium, Brucella mYcoides, Brucella anthracius, MYcobacterium ATCC_ 607, MvcQbacterium avium, Mvcobacterium Dhlei, Nocardia asteroides, SaccharomYces cervisiae, Candida albicans, Penicillium qlacum, and AsPergillus niqer.
The mitomycin derivatives of the present invention are useful in vitro as antiseptics, i.e. ~or disinfecting. The compounds are also useful top;cally and internally as therapeutic agents in combating pathogenic bacteria, e.g. in cases of staphylodermatitis, bacterial pneumoniae, leptopserosis, rickettsiosis, salmonellosis, and the like.
Typ;callyl for top kal application, the mitomycins of this invention are applied in compositions having concentrations in the range of 0.01 to l000 ug~ml.
As antineoplastic agents, the compounds of the present invention are useful in treating a variety of cancers, including, but not A68.2.WP 082288 -23- 1327~37 limited to, those cancers susceptible to cell growth suppression by the parent compounds. Treatment o~ cancers with the parent compounds are described in the following references:
Driscoll, J.S. ~t al., Cancer ChemotheraDY Rep. 4:1 (1974).
Kojima, R., et al., Cancer Chemotherauv Rep. 3:111 (1972).
Sugiura, K., Cancer Res. 19:438 (1959).
Oboshi, S., et al., Gann ~:315 (1967).
Sugiura, K., Cancer Chemotherapv Re~. 13:51 (1961).
Venditti, J.M., et al., Advances in Cancer ChemotheraDv, pp. 201-209 (1978) Editors: H. Umezawa et al., Japan Soc. Press, ~okyo/Univ.
Park Press, Balt;more.
Usubuchi, I., et al., ~Q 58:307 (1967).
Typical cancers treated by the mitomycin derivatives of this invention include, but are not limited to gastric and pancreatic neoplasms (Schein, P.S. et al., in M;tomYcin C: Curr*nt Status and New DeveloPments, pp. 133-143, Carter et al. Eds., Academic Press, New York (1979)~. Other cancers that may be treated using the compounds of the invention include lung, breast, anal, colorectal, head and neck, and melanoma.
The compounds of the invention are also active against the following tumor systems: Leukemia L-1210, Leukemia P388~ P1534 leukemia, Friend Virus Leukemia, Leukemia L4946, Mecca lymphosarcoma, Gardner lymphosarcoma, Ridgway Osteogenic sarcoma, Sarcoma 180 (ascites~, Wagner osteogenic sarcoma, Sarcoma T241, Lewis lung carcinoma, Carcinoma 7555 CD8F, Mammary Carcinoma, Colon 38, Carcinoma 1025, Ehrlich carcinoma (ascites & solid1, Krubs 2 carcinoma ~as-cites), Bashford carclnoma 63, Adenocarcinoma E 0771, B16 Melanoma, Hardin-Passey melanoma, 6~10m2 26, Miyona adenocarcinoma, Walker carcinosarcoma 256, Flexner-Jobling carcinoma, Jensen sarcoma, Iglesias sarcoma, Ig12sias ovarian tumor, Murphy-Sturn lymphosarcoma, Yoshida sarcoma, Dunning leukemia, Rous chicken sarcoma, and Crabb hamster sarcoma.

A68.2.WP ` 082288 ~ 24- ~327037 The pharmaceutical compositions of the present invention may be administered by any means that achieve their intended purpose. For example, admin;stration may be by parenteral, subcutaneous, intra-venous, intramuscular, intraperitoneal, transdermal, or buccal routes.
Alternatively, or concurrently, administration may be by the oral route. The dosage administered will be dependent upon the age, health, and weight of the recipient, kind of concurrent treatment, if any, frequency of treatment, and the nature of the effect desired.
Compositions within the ssope of this invention include all compositions wherein the mitomycin derivatiYe is contained in an amount effective to achieve its intended purpose. While individual needs vary, determ;nation of optimal ranges of effective amounts of each component is ~ith the skill of the art. Typical dosage forms contain 10 to 300 ~mole/kg animal of the mitomycin derivative, or an equivalent amount of the pharmaceutically acceptable salt thereof.
In addltion to the pharmacologically active compounds, the new pharmaceutical preparations may contain suitable pharmaceutically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. Preferably, the preparations, particu-larly those preparations which can be administered orally and which can be used for the preferred type of administration, such as tablets, dragees? and capsules, and also preparations which can be administered rectally, such as suppositories, as well as suitable solutions for administration by iniection or orally, contain from about n.ol to 99 percent, preferably from about 25 to 75 percent of active compound(s), together with the excipient.
The pharmaceutical preparations of the present invention are manufactured in a manner which is itself known, fur example, by means of conventional mixing, granulating, dragee-makiny, ~issolving, or lyophilizing processes. Thus, pharmaceutical preparatiuns for oral use can be obtained by combining the active cumpounds with solid excipients, optionally grinding the resulting mixture and processing A68.2.WP 082288 - ~ 1327~3~

the mixture of granulcs9 after adding suitable auxiliaries, if desired or necessary, to obtain tablets or dragee cores.
Suitable excipients are, in particular, fillers such as sac-charides, for example lactose or sucrose, mannitol or sorbitol, cellulose preparations and/or calcium phosphates, for example tri-calc;um phosphate or calcium hydrogen phosphate, as well as binders such as starch paste, using, for example9 ma ke starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropylmethylccllulose, sodium carboxymethylcellulose, and/or polyvinyl pyrrolidone. If desired, disintegrating agents may be added such as the above-mentioned starches and also carboxymethyl-starch, cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof, such as sodium alg;nate. Auxiliaries are, above all, flow-regulating agents and lubricants, for example, silica, talc, steric acid or salts thereof, such as magnesium stearate or calcium stearate, and/or polyethylene glycol. Dragee cores are provided with suitable coatings which, if desired, are res1stant to gastric juices. For this purpose, concentrated saccharide solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, poly-ethylene glycol and/or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. In order to produce coatings resistant to gastric juices, solutions of suitable cellulose preparations such as acetylcellulose phthalate or hydroxypropymethyl-cellulose phthalate, are used. Dye stuffs or pigments may be added to the tablets Dr dragee coatings, for example, for identification or in order to characterize combinations of active compound doses.
Other pharmaceutical preparations which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer such as glycerol or sorbitol. ~he push-fit capsules can contain the active compounds in the form of granules which may be mixed with fillers such as lactose, binders such as starches, and~or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active A68.2.WP 082288 --26- ~32~37 compounds are preferably dissolved or suspended in suitable liquids, such as fatty oils, or liquid paraffin. In addition, stabilizers may be added.
Possible pharmaceutical preparations which can be used rectally include, for example, suppos;tories, which consist of a combination of the active compounds with a suppository base. Suitable suppository bases are, for example, natural or synthetic triglycerides, or paraffin hydrocarbons. In addition, it is also possible to use gelatin rectal capsules which consist of a combination of the active compounds with a base~ Possible base materials include, for example, liquid triglycerides, polyethylene glycols, or paraffin hydrocarbons.
Suitable formulat;ons for parenteral administration include aqueous solutions of the active compounds in water-soluble form, for example, water-soluble salts. In addition, suspensions of the active compounds as appropriate oily injection suspensions may be adminis-tered. Suitable lipophilic solYents or vehicles include fatty oils, for example, sesame oil, or synthetic fatty acid esters, for example, ethyl oleate or triglycerides. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension include, for example, sodium carboxymethyl cellulose, sorbitol, and/or dextran. Optionally, the suspension may also contain stabilizers.
The following examples are illustrative, but not limiting, of the method and compositions of the present invention. Other suitable modifications and adaptations of the variety of conditions and parameters normally encountered in clinical therapy and which are obvious to those skilled in the art are within the spirit and scope of the invention.

A68.2.WP 082288 ~'~2~3~

EXAXPLES

Exam~le 1: Preparation of M;$omYcin A

Mitomycin C (50 mg, 0.15 mmol) was dissolved in 3 ml of a solution of 50% methanol and 50% 0.1 N NaOH and stirred at room temperature for 18 hrs. After completion of the reaction (TLC, CHCl3:MeOH, 10:1), the reaction mixture was quenched with dry ice to neutralize sodium hydroxide. The mixture was then freeze-dried in vacuo, and the mitosane compound was removed with methanol. The methanol solution was concentrated in vacuo to dryness, and thP
residue was redissolved in a minimum amount of methanol and then precipitated with ether to give 20 mg of a red-purplish powder. ~his was dissolved in 15 ml of ethyl acetate and cooled to 5~C, treated with diazomethane (etherial solution of diazomethane was prepared according to the procedure of Arndt, Ora. SYnthesis, Collective Volume Il, pp. 165-167~, and stirred for 20 minutes (TLC, CHCl3:MeOH, 10:1).
After completion of the reaction, the solvent was first removed under water aspirator reduced pressure and then dried in vacuo. The residue was recrystallized from ether to give 18 mg of reddish needles, m.p.
159-160~C. TLC (EtOAc:acetone, 1:1) gave one spot Rf = 0.91, and (acetone:benzene, 4:1) one spot R~ s 0.48. UV (methanol3 216 and 358 mu. NMR (acetone-d6, middle peak nf acetone at 2.10~, ~, 5.94 (br, 2H); 4.76 (dd, 1H~; 4.38 (t, lH); 4.07 (s, 3H~; 3.96 (d, lH)i 3.54 (dd, lH); 3.41 (d, lH); 3.35 (d, lH); 3.25 Is, 3H); 2.99-2.264 (mmm);
2.87 (s); 1.640 ~s, 3H).

Example 2: Prepar~tion of N7-(2-deoxvqlucopYranosyl)mitomvcin O

To a solution of mitomycin A ~10 mg, 0.028 mmol) in absolute methanol was added a methanolic solution of glucosamine HOl (70 mg, 0.325 mmol) and diisopropylethylamine ~100 ul~. This mixture was stirred under N2 atmosphere at room temperature until the reaction was A68.2.~P 082288 -28- ~ 3~3~

complete by TLC (EtOAc: acetone, 1 1) at which time (10 hrs.) the solution had changed color from reddish to dark purple. The solution was concentrated by evaporation with a N2 s~ream and chromatoyraphed on a preparative silica plate eluted with acetone-ethyl acetate (1:1).
The purple band remaining close to the origin was scraped off and eluted with methanol. Further purification by HPLC (C18 reversed phase, semi-preparative column, methanol: O.lN phosphate buffer, 1:1) gave a purple powder, NMR (D20) ~ 5.32 ~d, lH~ saccharide anomeric H~;
3.85 (s, 3H, 9a-OCH3); and the disappearance of singlet at 4.09 (Matsui7 M., et al., J. Antibiot. 21:189 (1968~; Cheng, L., and Remers, W.A., J. Med. Chem. 20:767 (1977); Vyas, D.M., e~ al., J. Ora.
Chem. 51:4307 (1986)).

Example 3: Preparation of N-(2~ DihvdroxYcYclohexyl~qlYcinamide To a solution of N-benzyloxycarbonylglycine (3 g, 14.3 mmol) in dioxane was added N-hydroxysuccinimide (1.65 9, 14.3 mmol) and N,N-dicyclohexylcarbodiimide ~2.96 y, 14.3 mmol) with cooling. The reaction mixture was stirred at 0-5C for one hour and allowed to stand under refrigeration overnight. The urea precipitate was removed by suction filtration and the filtrate was concentrated in vacuo to dryness. The yellowish residue was recrystallized from ethyl acetate-ether to give an 84% yield of the glycine activated ester mp. 112-114C. NMR ~CHC13).
The above-prepared activated ester of glycine (25 9, O.C08 mol) was dissolved in 15 ml of dry DMF (dimethyl formamide), chilled to below S-C, and 2-amino-l~3-cyclohexanediol (2.1~ 9, 0.016 mol) in DMF
was added drop-wise with stirring under N2 atmosphere. After comple-tion of the reaction (TLC, CHC13:MeOH~ , DMF was removed under reduced pressure and the result~ng solid residue was crystallized from ethyl acetate to give white crystals in 84% yield, m.p. 170-172~C.
NMR (D20): ~ 7.45 ~s, 5H, aromatic H); 5.20 (s, 2H, benzylic-CH2);
3.95 (s, 2H, -CO-CH2^NH2); 3.6 (t, lH, ClH of cyclohexane ring); 3.45 A68.2.WP 082288 ~L327~37 (m, 2H, C2H and C6H of cyclohexane ring); 2.0, 1.8 and 1.35 (m,m,m, 2 to 1 to 3H; C3, C4 and C5 hydrogens of cyclohexane). The product comprises the N-protected benzyloxycarbonyl derivative of N-(2,6-dihyroxycyclohexyl)glycinamide.
N-protected benzyloxycarbonyl N-(2,6-dihydroxycyclohexyl) glycinamide (39, 0.033 mol) was dissolved in 100 ml of absolute ethanol with a molar equivalent of 10% HCl. Hydrogenolysis with 5%
Pd/C at 30 psi, removal of the catalyst over celite, and subsequent evaporation of solvents in vacuo yielded a pale brownish solid which was triturated with ether and recrystallized from ethyl acetate and ether, m.p. 207-210C. NMR (320~ ~, 3.65 (t, 3H, ClH of cyclohexane ring); 3.55 (m, 2H, C2H and C6H of cyclohexane~; 3.4 (s, 2H, -C0-CH2-NH2); 2.05, 1.80, and 1.38 (m,m,m, 2 to 1 to 3H9 hydrogens of C3, C4 and C5 of cyclohexane~.

ExamDle 4 Animal Studies The compound N7-(2-deoxyglucopyranosyl)mitomycin C, prepared according to Example 2, was evaluated for both murine P388 leukemia antitumor activity and toxicity to bone marrow in normal mice.

A. Determination of murine antitumor actiyitY
The murine P388 leukemia system, maintained intraperitoneally in female DBA/2 mice, was used to evaluate antitumor activity. This tumor was selected because of its known sensitivity to the parent compound, mitomycin C (Driscoll ~ ancer ChemotheraDv ReDorts 4:1 (1974~). N7-(2-deoxyylucopyranosyl~mitomycin C was dissolved in sterile water (at 4-C) immediately prior to administration. Mitomycin C was dissolved in ethanol, and the resultant solution was adjusted to 5% ethanol, 95% sterile water.

A68.2.WP 082288 ~2~

Each compound was administered ;ntraperitoneally to groups of CD2F1 male mice on Day 1 after intraperitoneal implantation of 1 x 106 P388 leukemia cells. The P388 antileukemic activity of the test compound was assessed by mean survival days and percentage increased life span (ILS). The % ILS was calculated as follows:
%ILS ~ (T-C3/C x 100;
where ~ is the mean survival days of the treated mice and C is the mean survival days of the untreated mice.
P388 antitumor activi$y for N7-(2-deoxyglucopyranosyl)mitomycin C9 in comparison with the parent mitomycin C, is summarized in Table 2:
Table 2 Antitumor Activitv Against P388 Leukemia DruqDose (mg/kq~ %ILS Mean Survival (davs) N7-(2- Sa 42% 14.2 deoxygluco-pyranosyl) mitomycin C13.5a 61% 16.1 Mitomycin C 4.5b ~1% 18.1 Control C 10 . 0 aLDo dose bApproximate LD1~ dose CTreated with drug vehicle B. Determination of the effects of N7-(?-deoxYqlucoP.Yranosyl) mitomycin C on the hematopoietic svstem in mice Measurement of peripheral 7eukocyte (WBC) count was performed us;ng a 20-ul sample of retro-or~ital sinus blood obtalned from normal CD2F1 male ~ice on Day 3 ~ollowing i.p. administration of 13.5 mg/kg of N7-~2-deoxyglucopyranosyl~mitomycin C or 4.5 mg/kg of mitomycin C.
Afi8.2.WP 082288 _31_ ~327~37 Blood samples obtained were diluted in 9.98 ml of Iso~on (a neutral, isotonic buffer solution) and counted in a Coulter counter after lysis with Zapoglobin ~an enzyme solution which lyses red blood cells, but not white blood cells). WBC counts are expressed as a percentage of ~alues fro~ control mice receiving drug vehicle only. The results are summarized in the following table:
Table 3 In vivo Murine WBC DePression WBC Count on Day 3 Dose (as Dercent of control) N7-(deoxy- 13.5 mg/kg ~4%
glucopyranosyl) mitomycin C
Mitomycin C 4.5 mg/kg 56-66%

In summary, these in vivo studies demonstrate that N7-(2-deoxy-glucopyranosyl)mitomycin C has significant activity against the murine P388 tumor system, at doses producing no sign;f;cant bone marrow toxicity, as determined by depression of peripheral leukocyte ~WBC) count.

Example 5: Antibacterial Activitv N7-(2-deoxyglucopyranosyl)mitomycin C was evaluated for activity against Gram-negatiYe bacteria, in a comparative study with the parent mitnmycin C. Minimum inhibition concentration (M.I.C.~ against a Gram-negative strain of bacteria (HB101) was estimated by the dilution methnd, with graded concentrations of drug added to agar at 37-40C.
N7-(2-deoxyglucopyranosyl)m;tomycin C was dissolved in 50% sterile water-50% ethanol at 4-C, and mitomycin C was dissolved in ethanol.
The agar, containing drug, quickly solidified at room temperature, and A68.2.WP 082288 -32- ~3~37 the bacteria were plated immediately. After 24 hours at 37C, the agar plates were observed for inhibition of bacterial grow~h. The results are summarized in Table 4.
Table 4 M.l.C.
Compound Çram-neqative Bacteria N7-(2-deoxyglucopyranosyl) 1.66-3.3 ~cg/ml ~itomycin C
Mitomycin C 0O3 -0.5 mcg/ml .

Having now fully described this invention, it will be understood by those of skill in the art that the same can be pe-rformed within a wide and equivalent range of conditions, formulations and other parameters without affecting the scope of the invention or any embodiment thereof.

A68.2.~P 082288

Claims (35)

1. A mitomycin derivative having the formula:

wherein, n is 0 or 1;
Y is selected from the group consisting of glucopyranosyl, galactopyranosyl, mannopyranosyl, xylopyranosyl, cellobiosyl, lactosyl, glucofuranosyl, maltosyl, and 1,3-cyclohexanediol-2-yl, or the hydroxyl-protected acetate derivatives thereof;
R is hydrogen;
R1 is hydrogen, C1-C4 alkyl or C1-C4 alkyl substituted by phenyl, hydroxyphenyl, indolyl, mercapto, C1-C4 alkylthio, hydroxy, carboxy, amino, guanidino, imidazole or carbamyl; or R and R1 form a five or six membered ring containing nitrogen.

2. The mitomycin derivative of claim 1, comprising N7-(2-deoxyglucopyranosyl)mitomycin C.

3. The mitomycin derivative of claim 1, comprising N7-(2-deoxygalactopyranosyl)mitomycin C.

4. The mitomycin derivative of claim 1, comprising N7-(tetraacetyl-2-deoxyglucopyranosyl)mitomycin C.

5. The mitomycin derivative of claim 1, comprising N7-(tetraacetyl-2-deoxygalactopyranosyl)mitomycin C.

6. The mitomycin derivative of claim 1, comprising N7-[[[(tetraacetyl-2-deoxy-2-glucopyranosyl)amino]carbonyl]
methyl]mitomycin C.

7. The mitomycin derivative of claim 1, comprising N7-[[[(2-deoxyglucopyranosyl)amino]carbonyl]methyl]
mitomycin C.

8. A mitomycin derivative having the formula wherein, n is 0 or 1;
Y is selected from the group consisting of glucopyranosyl, galactopyranosyl, mannopyranosyl, xylopyranosyl, cellobiosyl, lactosyl, glucofuranosyl, maltosyl, and 1,3-cyclohexanediol-2-yl, or the hydroxy-protected acetate derivative thereof;
R is hydrogen;
R1 is hydrogen, C1-C4 alkyl or C1-C4 alkyl substituted by phenyl, hydroxyphenyl, indolyl, mercapto, C1-C4 alkylthio, hydroxy, carboxy, amino, guanidino, imidazole or carbamyl; or R and R1 form a five or six membered ring containing nitrogen; and R2 is NH2- or CH3O-.

9. The mitomycin derivative of claim 8, comprising N1-[[2-[[(2-deoxyglucopyranosyl)amino]carbonyl]ethyl]-carbonyl]
mitomycin C.

10. The mitomycin derivative of claim 8, comprising N1-[[2-[[(2-deoxyglucopyranosyl)amino]carbonyl]ethyl] carbonyl]mitomy-cin A.

11. A mitomycin derivative having the formula wherein, R2 is NH2- or CH3O-; and R3 is a 3-cyano-4-morpholinyl-2-deoxypyranosyl saccharide or a 4-morpholinyl-2-deoxypyranosyl saccharide.

12. The mitomycin derivative of claim 11, comprising 2-(3-cyano-4-morpholinyl)-2-deoxyglucopyranosyl-1a-carbothioamide mitomycin.

13. The mitomycin derivative of claim 11, comprising 2-(3-cyano-4-morpholinyl)-2-deoxygalactopyranosyl-1a-carbothioamide mitomycin.

14. The mitomycin derivative of claim 11, comprising 2-(4-morpholinyl)-2-deoxyglucopyranosyl-1a-carbothioamide mitomycin.

15. A pharmaceutical composition comprising the mitomycin derivative of any one of claims 1, 8 or 11 and a pharmaceutically acceptable carrier.

16. The use of an antibacterial amount of the mitomycin derivative of any one of claims 1, 8 or 11 and a pharmaceutically acceptable carrier for treating bacterial infection in an animal.

17. The use in accordance with claim 16, wherein said bacterial infection is caused by a bacteria selected from the group consisting of Escherichia, Pseudomonas, Salmonella, Staphylococcus, Klebsiella and Listeria.

18. The use of a cancer cell growth suppressing amount of the mitomycin derivative of any one of claims 1, 8 or 11, and a pharmaceutically acceptable carrier for treating cancer by suppressing growth of cancer cells susceptible to growth suppression in an animal.

19. The use in accordance with claim 18, wherein said cancer is selected from the group consisting of leukemia, melanoma, sarcoma, and carcinoma.

20. A process for preparing an N7-substituted mitomycin deriva-tive of the formula:

wherein, n is 0;
Y is selected from the group consisting of glucopyranosyl, galactopyranosyl, mannopyranosyl, xylopyranosyl, cellobiosyl, lacto-syl, glucofuranosyl, maltosyl, and 1,3-cyclohexanediol-2-yl, or the hydroxyl-protected peracetate derivative thereof;
comprising:
reacting mitomycin A with an amino compound under basic condi-tions in a polar organic solvent to give the N7-substituted mitomycin.

21. The process of claim 20, wherein said amino compound is selected from the group consisting of glucosamine, galactosamine, mannosamine, xylosamine, cellobiosamine, maltosamine, and 2-amino-1,3-cyclohexanediol.

22. The process of claim 20, wherein said N7-substituted mitomycin is N7-(2-deoxyglucopyranosyl) mitomycin C.

23. A process for preparing a mitomycin derivative having the formula wherein n is 1;

Y is selected from the group consisting of glucopyranosyl, galactopyranosyl, mannopyranosyl, xylopyranosyl, cello-biosyl, lactosyl, glucofuranosyl, maltosyl, 1,3-cyclohexane-diol-2-yl, or the hydroxyl-protected peracetyl derivative thereof;
R is hydrogen;
R1 is hydrogen, C1-C4 alkyl or C1-C4 alkyl substituted by phenyl, hydroxyphenyl, indolyl, mercapto, C1-C4 alkylthio, hydroxy, carboxy, amino, guanidino, imidazole or carbamyl; or R and R1 together form a five or six membered nitrogen containing ring;
comprising:
(a) condensing an N-protected amino acid with an alcohol in the presence of a dehydration reagent to give an activated ester, (b) condensing the activated ester obtained in step (a) with an amino compound to give a protected amino acid-amino compound con-jugate, (c) removing the amino acid protecting group of the protected amino acid-amino compound conjugate obtained in step (b) to give an amino acid-amino compound conjugate, and (d) condensing the amino acid-amino compound conjugate obtained in step (c) with mitomycin A to give the mitomycin derivative.

24. The process of claim 23, wherein said N-protected amino acid is selected from the group consisting of the N-benzyloxycarbonyl derivatives of alanine, valine, leucine, isoleucine, proline, phenyl-alanine, tryptophan, methionine, glycine, serine, threonine, cysteine, tyrosine, asparagine, glutamine, aspartic acid, glutamic acid, lysine, arginine and histidine.

25. The process of claim 23, wherein the amino acid protecting group is removed by hydrogenolysis.

26. The process of claim 20, wherein the base is a hindered amine selected from diisopropylethylamine, a C1-C3 trialkylamine, DBU or DMAP.

27. A process for the preparation of a mitomycin derivative having the formula:

wherein R2 is NH2- or CH3O-; and R3 is a 2-(3-cyano-4-morpholinyl)-2-deoxy pyranosyl saccharide;
comprising (a) condensing bis(acetaldehyde-2-yl) ether with a 2-amino-2-deoxy pyranosyl saccharide in the presence of a salt of cyanoborohydride to give a 2-deoxy-2-(3-cyano-4-morpholinyl) pyranosyl saccharide and a 2-deoxy-4--morpholinyl pyranosyl saccharide;
(b) separation of the 2-deoxy-2-(3-cyano-4-morpholinyl) pyranosyl saccharide from the 2-deoxy-4-morpholinyl pyranosyl saccharide obtained in step (a);
(c) reaction of the 2-deoxy-2-(3-cyano-4-morpholinyl) pyranosyl saccharide obtained in step (b) with an acetyl halide to give a 2-deoxy 1-halo-2(3-cyano-4-morpholinyl) peracetyl pyranosyl saccharide;

(d) treatment of the 2-deoxy-1-halo-2-(3-cyano-4-morpholinyl) peracetyl pyranosyl saccharide obtained in step (c) with silver thiocyanate to give a pyranosyl saccharide-1-thiocyanate;
(e) reaction of the pyranosyl saccharide-1-thiocyanate obtained in step (d) with mitomycin C or mitomycin A to give a mitomycin C- or mitomycin A-pyranosyl saccharide peracetate carbothioamide; and (f) hydrolysis of the acetate groups of the mitomycin-C-pyranosyl saccharide peracetate obtained in step (e) to give the mitomycin derivative.

28. A process for the preparation of a mitomycin derivative having the formula:

wherein R2 is NH2- or CH3O-; and R3 is a (4-morpholinyl)-2-deoxy pyranosyl saccharide;
comprising (a) condensation of bis(acetaldehyde-2-yl) ether with a 2-amino-2-deoxy pyranosyl saccharide in the presence of a salt of cyanoborohydride to give a 2-deoxy-2-(3-cyano-4-morpholinyl) pyranosyl saccharide and a 2-deoxy-2-(4-morpholinyl) pyranosyl saccharide;
(b) separation of said 2-deoxy-2-(4-morpholinyl) pyranosyl saccharide from said 2-deoxy-2-(3-cyano-4-morpholinyl) pyranosyl saccharide obtained in step (a);

(c) reaction of the 2-deoxy-2-(4-morpholinyl) pyranosyl saccharide obtained in step (b) with an acetyl halide to give a 2-deoxy-1-halo-2-(4-morpholinyl) peracetyl pyranosyl saccharide;
(d) treatment of the 2-deoxy-1-halo-2-(4-morpholinyl) peracetyl pyranosyl saccharide obtained in step (c) with silver thiocyanate to give a pyranosyl saccharide-1-thiocyanate;
(e) reaction of the pyranosyl saccharide-1-thiocyanate obtained in step (d) with mitomycin A or C to give a mitomycin A- or C-pyranosyl saccharide peracetate carbothioamide; and (f) hydrolysis of the acetate groups of the mitomycin-C-pyranosyl saccharide peracetate obtained in step (e) to give the mitomycin derivative.

29. The process of claim 28 or 29, wherein said 2-amino-2-deoxy pyranosyl saccharide is selected from the group consisting of glucosamine, galactosamine, mannosamine, xylosamine, cellobiosamine and maltosamine.

30. The process of claim 28, wherein said mitomycin derivative is 2-(3-cyano-4-morpholinyl)-2-deoxyglucopyranosylmitomycin-1a-carbothioamide.

31. The method of claim 29, wherein said mitomycin derivative is 2-(4-morpholinyl)-2-deoxyglucopyranosyl-mitomycin-1a-carbothioamide.

32. A process for the preparation of a mitomycin derivative having the following formula wherein n is 0;
Y is selected from the group consisting of glucopyranosyl, galactopyranosyl, mannopyranosyl, xylopyranosyl, cello-biosyl, lactosyl, glucofuranosyl, maltosyl, 1,3-cyclohexane-diol-2-yl;
R is hydrogen;
R1 is hydrogen, C1-C4 alkyl or C1-C4 alkyl substituted by phenyl, hydroxyphenyl, indolyl, mercapto, C1-C4 alkylthio, hydroxy, carboxy, amino, guanidino, imidazole or carbamyl; or R and R1 together form a five or six membered nitrogen containing ring;
R2 is NH2- or CH3O;
comprising:
(a) condensation of mitomycin C with succinic anhydride to give mitomycin C-1a-succinic acid ester;
(b) condensation of the mitomycin C-1a-succinic acid ester obtained in step (a) with a hydroxyl-protected amino derivative selected from the group consisting of glucosamine, galactosamine, mannosamine, xylosamine, cellobiosamine, maltosamine, and 2-amino-1,3-cyclohexanediol; and (c) removal of the hydroxyl protecting groups to give the mitomycin derivative.

33. A process for the preparation of a mitomycin derivative having the following formula wherein n is O;
Y is selected from the group consisting of glucopyranosyl, galactopyranosyl, mannopyranosyl, xylopyranosyl, cellobiosyl, lacto-syl, glucofuranosyl, maltosyl, 1,3-cyclohexanediol-2-yl;
R is hydrogen;
R1 is hydrogen, C1-C4 alkyl or C1-C4 alkyl substituted by phenyl, hydroxyphenyl, indolyl, mercapto, C1-C4 alkylthio, hydroxy, carboxy, amino, guanidino, imidazole or carbamyl; or R and R1 together form a five or six membered nitrogen containing ring;
R2 is NH2- or CH3O-;
comprising:
(a) condensation of mitomycin A with succinic anhydride to give mitomycin A-1a-succinic acid ester;
(b) condensation of the mitomycin A-1a-succinic acid ester obtained in step (a) with a hydroxyl-protected amino acid-saccharide conjugate of the formula wherein R, R1 and n are as defined above and YP is a hydroxyl-pro-tected saccharide selected from the group consisting of the hydroxyl-protected derivatives of glucopyranosyl, galactopyranosyl, manno-pyranosyl, xylopyranosyl, cellobiosyl, lactosyl, glucofuranosyl, maltosyl, and 1,3-cyclohexanediol-2-yl; and (c) removal of the hydroxyl protecting groups to give the mitomycin derivative.

34. The use of an antibacterial amount of the mitomycin derivative of any one of claims 1, 8 or 11 for the preparation of a medicament for treating bacterial infection in an animal.

35. The use of a cancer cell growth suppressing amount of the mitomycin derivative of any one of claims 1, 8 or 11, for the preparation of a medicament for treating cancer by suppressing growth of cancer cells susceptible to growth suppression in an animal.