NZ206932A - Certain 6-(heterocyclyl or amino) mitosanes and pharmaceutical compositions - Google Patents

Description

New Zealand Paient Spedficaiion for Paient Number £06932 206932 No.: Date: "i-fity D?" Complete Specification tiieo: Class: .CffWl&Ijtfy B ay • • ■ .. • | y -AUB • ? Publication Date; I, No: .l?rC|Cj P.O. Journal, NEW ZEALAND PATENTS ACT, 1953 v C f N' COMPLETE SPECIFICATION MITOMYCIN ANALOGS ?/ We, UNIVERSITY PATENTS, INC., a Delaware corporation, of 537 Newtown Avenue, Norva.lk, Connecticut 06851 , United States of America, hereby declare the invention for which ^ / we pray that a patent may be granted to ixqe/us, and the method by which it is to be performed, to be particularly described in and by the following statement: - (followed by page la) 208932 - iq- "MITOMYCIN ANALOGS" BACKGROUND The present invention relates generally to antibiotic mitosane compounds and to their use in the treatment of neoplastic disease states in animals.

The disclosures of my U.S. Letters Patent No. 4,268,676; my U.S. Patent 4,460,599; and my NZ Patent Specification No. 199,617, - - . I are specifically incorporated by reference herein to the extent that they may provide essential and nonessential material relating to the present invention.

Briefly summarized, said U.S. Patents Nos. 4,268 ,676 and 4,460,599 : set forth a statement of the background of the ongoing search in the art for new and useful compounds which are structurally related to the mitomycins, which possess antibiotic activity, which have low toxicity and which display a substantial degree of antitumor activity in animals. More particularly, they disclose new compounds of the formula I, O ch2ocnh2 OCH- 3 -IcJ wherein: Y is hydrogen or lower alkyl; and X is a thiazolamino radical, a furfurylamino radical o^ radical of the formula, 206932 R I N- - 2 -1 1 2 in which R, R and R are the same or different and selected from the group consisting of hydrogen and lower alkyl, and R3 is selected from the group consisting of lower alkenyl, halo-lower alkenyl, lower alkynyl, lower alkoxycarbonyl, thienyl, formamyl, tetrahydrofuryl and benzene sulfonamide.

The term "lower" as used herein to qualify the terms alkyl, alkenyl, alkynyl, alkaxy, alkoxy carbonyl and alkylamino means a carbon-containing substituent of frcm 1 to 6 cation atcms.

Said US patents also disclose novel methods for treatment of neoplastic disease states in animals, which methods comprise administering a therapeutically effective amount of a compound of the formula, la, ,CH2OCNH2 rfe 1 1 I 1 r~ga OCH3 la ;NY 3-la wherein: Y is hydrogen or lower alkyl; and 2 is a thiazolamino radical, a furfurylamino radical, a cyclopropylamino radical, a pyridylaminc radical, or a radical of the formula/ R' i — N" R" I C- •R .6 - 206932 4 5 6 in which R , R , and R are the sajne or different and selected from the group consisting of hydrogen and 7 lower alkyl, and R is selected from the group consisting of lower alkenyl, halo-lower alkenyl, lower alkynyl, 5 lower alkoxycarbonyl, halo-lower alkyl, hydroxy-lower alkyl, pyridyl, thienyl, formamyl, tetrahydrofuryl, benzyl, and benzene sulfonamide.

New Zealand Patent Specification No. 199,617 also discloses compounds with a substantial 10 degree of antitumor activity in animals of the following formula Ila, Ila 3 ~la wherein: Y is hydrogen or lower alkyl; and Z Is a lower alkoxy substituted quinolinylamino radical, a cyano substituted pyrazolylamino radical or a mono-or di-lower alkyl substituted thiazolamino radical, or a nitrogen-containing heterocyclic radical 25 selected from the group consisting of 1-pyrrolinyl, 1-indolinyl, N-thiazolidinyl, N-morpholinyl, 1-piperazinyl, and N-thiomorpholinyl radicals, or a cyano, phenyl, carboxamido or lower alkoxycarbonyl substituted 1-aziridinyl radical, or a lower alkyl, formyl or acetylphenyl substituted 1-piperazinyl radical, or an hydroxy or piperidyl substituted l-pip^rl&^l( radical, or . a lower alkoxy, amino or halo siabs-titute^^ NOV 1986 35 pyridylamino radical, or V 206932 a carboxamido, mercapto or methylenedioxy substituted anilino radical, or R I a radical of the formula,, -N-R1 5 wherein R is hydrogen or lower alkyl and R' is a nitrogen-containing heterocyclic radical selected from the group consisting of quinuclidinyl, pyrazolyl, 1-triazolyl, isoquinolinyl, indazolyl, benzoxazolyl, thiadiazolyl and benzothiadiazolyl, and lower alkyl and 10 halo substituted derivatives thereof, or a butyrolactonyl radical, or an adamantyl radical, or a mono-lower alkoxy substituted phenyl radical, or a substituted lower alkyl radical selected from the group consisting of mercapto lower alkyl, carboxy lower alkyl, mono-, di- and tri-lower alkoxy lower alkyl, lower alkyl thio lower alkyl and lower alkoxycarbonyl substituted derivatives thereof, cyano 20 lower alkyl, mono-, di- and tri-lower alkoxy phenyl lower alkyl, phenyl cyclo lower alkyl, 1-pyrrolidinyl lower alkyl, N-lower alkyl pyrrolidinyl lower alkyl, N-morpholinyl lower alkyl, and lower dialkylamino lower alkyl.

Also pertinent to the background of the present invention are the following references: Cosulich, et al., U.S. Patent No. 3,332,944 and New Zealand Patent Specification No. 129764; Matsui, et al., U.S.

Patent No. 3,410,867; Nakano, et al., U.S. Patent No. 4,231,936; Matsui, et al., U.S.. Patent No. 3,429,894; 30 Remers, U.S. Patent No. 4,268,676; Matsui, et al., U.S.

Patent No. 3,450,705; Matsui, et al., U.S. Patent No. 3,514,452; and Imai, et al., Gann, 71, pp. 560-562 ,/a I (1980). / V '7 NOV 1986 V 206933 BRIEF SUMMARY According to the present invention, there are provided novel compounds of the formula, III, ^ CH0OCNH0 s-*-- ^ III -1 a is or or wherein: Y is hydrogen or lower alkyl; and X an hydroxy substituted 1-pyrrolidinyl radical, a lower alkyl substituted 1-piperidyl radical, an acetamino, acetyl, carbamido, cyano, ~carboxy lower alkylamino, di-lower alkoxy, nitro, or sulfamyl substituted anilino radical, or R I i a radical of the formula, -N-R 2 5 wherein R is hydrogen or lower alkyl and R"*" is a nitrogen containing heterocyclic radical selected from the group consisting of amino substituted triazolyl, lower alkyl substituted isothiazolyl, benzothiazolyl, and nitro and halo substituted derivatives of benzo-30 thiazolyl, or R1 is a substituted lower alkyl radical selected from the group consisting of amino lower alkyl, lower alkylamino lower alkyl, hydroxy lower alkylamino lower alkyl, hydroxy lower alkoxy lower alkyl, imidazolyl 35 lower alkyl, nitro substituted imidazolyl lower al.|^l", (17 NOV 1986 7 2Q6$32 mong- and di-hydroxy phenyl lower alkyl, nitro substituted pyridylamino lower alkyl, and piperazinyl lower alkyl.

Also provided according to the invention are 5 novel methods for treatment of neoplastic disease states in animals, which methods comprise administering a therapeutically effective amount of a compound of the formula, Ilia, 7c, '6 1 | 1 I r-Sa.

Ilia ^NY 2. S—la wherein: Y is hydrogen or lower alkyl; and Z or or an hydroxy substituted 1-pyrrolidinyl radical, a lower alkyl substituted 1-piperidyl radicals a 1-piperazinyl radical or an acetamino, acetyl, carbamido, cyano, carboxy lower alkylamino, di- lower alkoxy, nitro, sulfamyl, or lower alkyl substituted anilino radical, or a radical of the formula, -N-R wherein R is hydrogen or lower alkyl and R"*" 30 is a, nitrogen containing heterocyclic radical selected from the group consisting of amino substituted tri-azolyl, lower alkyl substituted isothiazolyl, benzo-thiazolyl, and nitro and halo substituted derivat^.; of benzothiazolyl, or R^ j-s %• a substituted lower alkyl radical select^ from the group consisting of amino lower alkyl, lower 206932 alkylamino lower alkyl, hydroxy lower alkylamino lower alkyl, hydroxy lower alkoxy lower alkyl, imidazolyl lower alkyl, nitro substituted imidazolyl lower alkyl, mono- and di-hydroxy phenyl lower alkyl, nitro sub-5 stituted pyridylamino lower a,lkyl, piperazinyl lower alkyl, and pyridyl ethyl.

Unless otherwise indicated, the term "lower", applied to "alkyl" radicals shall designate such straight or branched chain radicals as to include from one to 10 six carbon atoms. By way of illustration, "lower alkyl" shall mean and include methyl, ethyl, propyl, butyl, pentyl and hexyl radicals as well as isopropyl radicals, t-butyl radicals and the like. Similarly, "lower" as applied to "alkoxy" shall designate a radical 15 having one to six carbon atoms.

It will be apparent that the compounds of formula III are all comprehended by the specifications of formula Ilia. Put another way, all the novel antibiotic mitomycin derivatives of formula III are useful in 20 practice of the novel antineoplastic therapeutic methods which involve administration of compounds of formula Ilia.

Mitomycin derivatives of the invention are prepared by the reaction of mitomycin A with the appro-25 priately selected amine compounds. The N-alkylmitomycin (e.g., N-methylmitomycin) derivatives are similarly prepared by the reaction of a selected amine with N-alkylmitomycin A prepared from mitomycin C, e.g., according to the methods generally disclosed in Cheng, 30 et al., J.Med.Chem., 20,. No. 6, 767-770 (1977). The preparative reactions generally yield the desired product a,s a crystalline solid which is readily soluble in alcohol.

Therapeutic methods of the invention comprehend 35 the administration of effective amounts of one or more of the compounds of formula Ilia, as an active ingredient, 205932 together with desired pharmaceutically acceptable diluents, adjuvants and carriers, to an animal suffering from a neoplastic disease state. Unit dosage forms of compounds administered according to. the methods of the 5 invention may range from about 0.001 to about 5.0 mg and preferably from about 0.004 to about 1.0 mg, of the compounds. Such unit dosage quantities may be given to provide a daily dosage of from about 0.1 to about 100 mg per kg, and preferably from about 0.2 to about 51.2 10 mg per kg, of body weight of the animal treated.

Parenteral administration, and especially intraperitoneal administration, is the preferred route for practice of the inventive methods.

Other aspects and advantages of the present 15 invention will become apparent upon consideration of the following description.

DESCRIPTION OF INVENTION The following Examples 1 through 32, describing preparation of certain presently preferred compounds according to the invention, are for illustrative purposes only and are not to be construed as limiting the invention. Unless otherwise indicated, all reactions were carried 25 out at room temperature (20°C), without added heat.

Unless otherwise indicated, all thin layer chromatographic (TLC) procedures employed to check the progress of reactions involved the use of a pre-coated silica-gel plate and a mixture of methanol and chloroform (2:8 30 by volume) as a developing solvent.

Example 1 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-roethoxy-5-methyl-6-(3-hydroxy-l-pyrrolidinyl)-azirino[2' , 3 ':3/4] pyrrolo[1,2-aJ indole-4,7-dione carbamate A solution of mitomycin A (50 mg) in 6 ml of anhydrous methanol was treated with 3-pyrrclidinol (13 mg) under nitrogen at room temperature. Vfhen thin- layer chromatography on silica gel (2:8 methanol- chloroform as solvent) showed that starting material no longer was present, the mixture was filtered and evaporated under reduced pressure. The residue was purified by preparative thin-layer chromatography using the same solvent system. This procedure gave 23 mg (40% yield) of the desired product having a melting point of 82°- 85°C (decomposition) and providing the following analysis: NMR (DMSO-dc, TS): 'o' values in ppm. b Absence of the 6-methoxy peak at 4.02 and the appearance of new peaks at 1.6-2.2 (m,2), 2.8-3.1 (broad s,5) and 4.0-4.3 (m,l).

Example 2 - 1^s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- (3-methylpiperidin-l-yl) -azirino f2',3',:3,41 pyrrolo [1,2-a]indole-4,7-dione carbamate This compound was prepared by the procedure described in Example 1. From 70 mg of mitomycin A and 200 mg of 3-methyl piperdine was obtained 46 mg (55% yield) of the desired product haying a melting point of 30 75°-88°C (decomposition) and providing the following analysis: NMR (CDCl^, TS) : *51. values in ppm.

Absence of the 6-methoxy peak at 4.02 and the appearance of new peaks at 0.85 (d,3), 1.10-2.15 Cm,5) and 2.15-3.32 (m,4).

Example 3 1, s-la, 2, s-8, r~8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-~5-methyl-6-(1-piperazinyl)-azirino[2',3';3,4]pyrrolo-[1,2-a]indole-4,7-dione carbamate This compound was prepared by the procedure described in Example 1. From 60 mg of mitomycin A and mg of anhydrous piperazine was obtained 23 mg (34% yield) of the desired product having a melting point greater than 200°C (decomposition) and providing the following analysis: NMR (DKSO-d,, , TS): 'o1 values in ppm. o Absence of the 6-methoxy peak at 4.02 and the appearance of new peaks at 1.9 (broad s,l) 20 and 2.9 (s,8).

Example 4 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy- -methyl-6-[4-(acetylamino)aniljno]-azirino[2 ' , 3 ' : 3,4] " pyrrolo[1r2-a]indole-4,7-dxone carbamate This compound was prepared by the procedure described in Example 1. From 100 mg of mitomycin A and excess 4-(acetylamino) aniline wa,s obtained 102 mg (76% 30 yield) of the desired product having a melting point of 143°-145°C (decomposition) and providing the following analysis: «i .) NMR (CDCl^r TS) : '5' values in ppm. pr) Absence of the 6-methoxy peak at 4.02 and the !-\p > appearance of new peaks at 2.1 [s,3], 7.4 <- (d, 2), 7.6 (s,l) and 8.9-9.3 (s,l).

Example .5 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6-[3-(acetylamino)anilinoj-azirino[2',3':3,4] pyrrolo[1,2-a]indole-4,7-dione carbamate This compound was prepared by the procedure described in Example 1, except that a small amount of solid potassium carbonate was added. From 70 mg of mitomycin A and 150 mg of 3-(acetylamino)aniline was 15 obtained 67 mg (72% yield) of the desired product having a melting point of 140°-143°C (decomposition) and providing the following analysis: NMR (Acetone-d-, TS): 'o1 values in ppm. o Absence of the 6-methoxy peak at 4.02 and the 20 appearance of new peaks at 2.1 (s,3), 6.7-7.5 (m,4), 8.0 (broad s,l) and 9.3 (s,l).

Example 6 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6-(4-acetylanilino)-azirino[2',3':3,4j pyrrolo [1,2-a]indole-4,7-dione carbamate This compound was prepared by the procedure 30 described in Example 1, except that a small amount of solid potassium carbor.a,te was $dded. From 70 mg of mitomycin A and 510 mg of 4-acetvlaniline was obtained 25 mg (28% yield) of the desired product having a melting point of 103°-104°C (decomposition) and providing 35 the following analysis: NMR (CDC1_, TS) : '5 ' Values in, ppn\.

Qs- Absence of the 6-methoxy peak at 4.02 and the appearance of new peaks at 2.1 (s,3), 6.6 (d,2), 7.3 Cd/2) and 7.0-7.3 (broad s,l).

O , - "Wo Example -7 1ts-la,2,s-8,r-8a,s-8b-Hexahydro-8-(hydroxymethyl)-8a-methoxy-5-methyl-6-[4-(1-ureido)anilino]-azirino[2', I 1:3,4]pyrrolo [1,2-a]indole-4,7-dione carbamate This carbamiao-substituted compound was prepared by the procedure described in Example 1. From 50 mg of mitomycin A and 227 mg of 4-(1-ureido)aniline was obtained 49 mg (67% yield) of the desired product having a melting 15 point of 93°-95°C (decomposition) and providing the following analysis: NMR (CDCl^/ TS): 'o' values in ppm.

Absence of the 6-methoxy peak at 4.02 and the appearance of new peaks at 5.0 3 (s,2), 6.9 20 (d,2) , 7.3 (d, 2) , 8.0 (s,l) and 8.4 (s,l).

Example 8 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy- 5-methvl-6- (4-cyanoanilino) -azirino [ 2 ' , 3 ' : 3,4] pyrrolo [l~, 2~a] 25 indole-4,7-dione carbamate This compound wa,s prepared by the procedure described in Example 1, except that a small amount of solid potassium carbonate wa,s added. From 70 mg of 30 mitomycin A and 472 mg of 4-aminobenzonitrile wa,s obtained 23 mg (24% yield} of the desired product having a melting point of 124°-126°C (decomposition) and providing the following analysis: 0 13 O O NMR (CDC1-, TS): ' <5' values in ppm. 3 Absence of the 6-methoxy peaJc at 4.02 and the appearance of new peaks at 6.6 (d,2), 7.4 (d,2) and 7.0-7.3 (broad s,l).

Example 9 1rs-la,2,s-8,r-8a,s-8b-Hexahydro-8-(hydroxymethyl)-8a-methoxy-5-methyl-6-(3-cyanoanilino)-azirino(2', 3 ':3,4]pyrrolo[1,2-a] indole-4,7-dione carbamate described in Example 1, except that a small amount of solid potassium carbonate was added. From 71 mg of mitomycin A and 500 mg of 3-aminobenzonitrile was 15 obtained 30 mg (34% yield) of the desired product having a melting pcint of S7°-98°C (decomposition) and providing the following analysis: 1,s-la,2,s-8,r-8a,s-8b-Hexahydro-8-(hydroxymethyl)-8a-methoxy-5-25 methyl-6-[4-(N-glycyl)anilino]-azirino[2',3':3,4]pyrrolo [1,2-a]indole-4,7-dione carbamate described in Example 1. From 50 mg of mitomycin A and 249 mg of 4-(N-glycyl)aniline was obtained 62 mg (90% 30 yield) of the desired product having a melting point of 83°-85°C (decomposition) and providing the following analysis: This compound was prepared by the procedure NMR (CDCl^f TS) : '<5' values in ppm.

Absence of the 6-methoxy peak at 4.02 and the appearance of new peaks at 7.2-7.8 (m,4).

Example 10 This compound was prepared by the procedure NMR (DMSO-dg, TS): '6' values in ppm-Absence of the 6-methoxy peak at 4.02 and the appearance of new peaks at 3.1 (s,2), 6.3-6.6 (broad s,2), 6.6-6.8 (broad s,2) and 6.6-7.1 5 (broad s , 2) .

Example 11 1, s-la ,2, s-8, r-8a, s-8b-Hexahvdro-8- (hvdroxvmethvlj-8a-methoxy-5 -~met.hyl-6- ( 3 , 4-axmetiioxyanilino) -azirino [2 ' , 3 ' : 3 , 4 ] pyrrolo 10 [1,2-a]indole-4,7-dione carbamate This compound was prepared by the procedure described in Example 1. From 50 mg of mitomycin A and 229 mg of 3,4-dimethoxyaniline was obtained 61 mg (91% yield) 15 the desired product having a melting point of 114°-116°C (decomposition) and providing the following analysis: NMR (CDCl^, TS): 161 values in ppm.

Absence of the 6-methoxy peak at 4.02 and the 20 appearance of new peaks at 3.8 (s,6), 6.3-6.9 (m,3) and 7.7 (s,l). t - Example 12 1,s-la,2,s-8,r-8a,s-8b-Hexahydro-8-(hydroxymethyl)-8a-methoxy-5- j metnyl-6-(3,5-dimethoxyanilino)-azirino[21,3 'T3,4]pyrrolo [1,2-a]indole-4,7-dione carbamate This compound was prepared by the procedure described in Example 1. From 50 mg of mitomycin A and 30 229 mg of 3,5-dimethoxyaniline was obtained 60 mg (88% yield) of the desired product having a melting point of 98°-100°C (decomposition) and providing the following analysis: NMR (CDC13, TS): '6' values'in ppm.

Absence of the 6-methoxy peak at 4.02 and the _ N o appearance of new peaks at 3.8 (s,6), 5.9-6.4 Oq (broad s,3) and 7.6 (s,l).

Example 13 v 1, s-la ,2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-S -methyl-6- (4-nitroanilino) -azirino [2 ' , 3 ' : 3, 4 J pyrrolo 1.1,2-a] indole-4,7-dione carbamate This compound was prepared by the procedure described in Example 1, except that a small amount of solid potassium carbonate was added. From 70 mg of mitomycin A and 276 mg of 4-nitroaniline was obtained 16 mg (9% yield) of the desired product having a melting point of 132°-134° C (decomposition) and providing the following analysis: NMR (Acetone-d^, TS): '6' values in ppm. b Absence of the 6-methoxy peak at 4.02 and the appearance of new peaks at 6.9-7.3 (d,2), 7.4-7.9 (d,2) and 7.9-8.4 (broad s,l).

Example 14 1, s-la, 2, s-8 / r-8a, s-8b-Hexahydro~8- (hydroxymethyl) -8a-methoxy- 5 -methyl-6-(4-sulfamyianilino)-azirino[2',3':3,4]pyrrolo [1,2-a]indole-4,7-dione carbamate This compound was prepared by the procedure described in Example 1, except that a small amount of solid potassium carbonate was added. From 70 mg of mitomycin A and 6 88 mg of sulfanilamide was obtained 25 30 mg (26% yield) of the desired product having a melting point of 113°-115°C (decomposition) and providing the following analysis: NMR (CDCl^ / TS) : '<$' values in ppm.

Absence of the 6-methoxy peak at 4.02 and the 35 appearance of new peaks at 7.0 (d,2),' 7.5 (s,l) and 7.9 (d,2). ■•v *1-JUL1987 Example 15 3 1, s-la, 2, s-8 , r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxv- 5-methyl-6-(4-methylanilino)-azirino[2', 3 ':3,4 J pyrrolo[1,2-a] indole-4,7-dione carbamate This compound was prepared by the procedure described in Example 1. From 60 mg of mitomycin A and excess 4-methylaniline was obtained 63 mg (86% yield) of the desired product having a melting point of 113°-10 115°C (decomposition) and providing the following analysis: NMR (CDCl^r TS) : ' <5 1 values in ppm.

Absence of the 6-rnethoxy peak at 4.02 and the appearance of new peaks at 2.3 (s,3), 6.5-7.3 15 (broad s,4) and 7.6 (broad s,l). 1 _ "<>;p Example 16 1,s-la,2,s-8,r-8a,s-8b-Hexahydro-8-(hydroxymethyl)-8a-methoxy-5-2o methyl-6-(3-methylanilino)-azirino[2 ' , 3 ':3,4]pyrrolo[1,2-a] indole-4,7-dione carbamate This compound, was prepared by the procedure described in Example 1. From 70 mg of mitomycin A and 276 mg of 3-methylaniline was obtained 66 mg (78% yield) of the desired product having a melting point of 89°-91°C (decomposition) and providing the following analysis: NMR (CDCl^/ TS): '6' values in ppm.

Absence of the 6-methoxy peak at 4.02 and the appearance of new peaks at 2.4 (s,3), 6.7-7.5 (in, 4) and 7.8 (s, 1) .

Example 17 1, s-la, 2, s-8, r-8a, s-8t>-Hexahydro-8- (hydroxymethyl) -8a-methoxy- 5-methyl-6-[(5-amino-l,2,4-triazol-3-yl)amino]-azirino [2',3':3,4]pyrrolo [1,2-a]indole-4,7-dione carbamate 12 V-JUL1987" ' t? E f 'iP' « * t\ »; This, compound was prepared by the procedure ""•0 described in Example 1, except that a small amount of ^7'fo solid potassium carbonate was added. From 50" mg of mitomycin A and 30 mg of 3,5-diamino-l, 2,4-triazole was obtained 13 mg (.5.5% yield) of the desired product having a melting point of 117°-120°C (decomposition) and providing the following analysis: NMR (DMSO-d-r TS): 'q' values in pom. 6 Absence of the 6-methoxy peak at 4.02 and the 10 appearance of a new peak at 5.37 (s,3).

Example 18 1,s-la,2,s-8, r-8a,s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy- 5~ methyl-6-[(3-methylisothiazol-5-yl)aminoJ-azirino[2',5': J, 4] pyrrolo[1,2-a]indole-4,7-dione carbamate This compound was prepared by the procedure described in Example 1, except that 0.5 ml of triethylamine was added. From 60 mg of mitomycin A and 30 mg of 5-2 0 amino-3-methyl'iscthiazole hydrochloride was obtained 4.5 mg (8.5% yield) of the desired product having a melting point of 87°-90°C (decomposition) and providing the following analysis: NMR (CDCl^r TS): '6' values in ppm. 25 Absence of the 6-methoxy peak at 4.0 2 and the appearance of new peaks at 2.3 (s,3), 6.1 (s, 1) and 6.4 (s,1).

Example 19 1, s-la, 2, s-8, r-8a, s-8b-Hexahydr o-8- (hydroxymethyl) -8a-methoxv-5 -methyl-6 - [ (2-benzothiazolyl) amino] -azirino [2 ' , 3 ' : 3, 4 ] pyrrolo [1,2-a]indole-4,7-dione carbamate This compound was prepared by the procedure described in Example 1, except that a small "amount of solid potassium carbonate was added. From 50 mg of mitomycin A and 25 mg of 2-aminobenzathiazo.le was obtained 12 mg (18% yield) of the desired product having a melting point of 82°-85°C (decomposition) and providing the following analysis: 5 NMR (CDC13, TS): 'o' values in ppm.

Absence of the 6-methoxy peak at 4.02 and the appearance of new peaks at 7.1-8.0 (m,5).

Example 20 — i n 1, s-la, 2, s-8, r-8a, s-8b-Hexahvdro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6-[(6-nitr0ben20thiaz0l-2-yl)amino]-azirino [2',3':3,4]pyrrolo [1,2-a]indole-4,7-dione carbamate This compound was prepared by the procedure described in Example 1, except that a small amount of solid potassium carbonate was added. From 50 mg of mitomycin A and 30 mg of 2-amino-6-nitrobenzothiazcle was obtained 20 mg (27% yield) of the desired product having a melting point of 86°-89°C (decomposition) and providing the following analysis: NMR (DMSO-d,., TS): 'o' values in ppm. — b Absence of the 6-methoxy peak at 4.02 and the appearance of new peaks at 6.9-8.3 (nr, 4).

Example 21 1, s-la,2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy- 5-methyl-b-[(4-chiorobenzothiazol-2-yl)amino]-azirino [21,3':3,4]pyrrolo[1,2-a]indole-4,7-dione carbamate This compound was prepared by the procedure described in Example 1, except that a small amount of solid potassium carbonate was added. From 150 mg of mitomycin A and 27 mg of 2-amino- (4-chloro) -benzothiazole was obtained 30 mg (14% yield) of the desired product 35 having a melting point of 89°-91°C (decomposition) and providing the following analysis: .1 NMR (CDCl^/ TS): '5' values in ppm-Absence of the 6-methoxy peak at 4.02 and the appearance of new peaks at 7.1-8.0 Cbroad s, 4) .

Example 22 1,s-la,2,s-8,r-8a,s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy- 5-methy1-6-[(2-aminoethyl)aminoJ-azirino[2',5 ' : 3,4]pyrrolo [1,2-a]indole-4,7-dione carbamate : This compound was prepared by the procedure described in Example 1, except that the solvent was dichloromethane. From 50 mg of mitomycin A and 10 mg of 1,2-diaminoethane was obtained 35 mg (6 5% yield) of 15 the desired product having a melting point of 202°-205°C (decomposition) and providing the following analysis: NMR (CDCl^, TS): 'o' values in ppm.

Absence cf the 6-methoxy peak at 4.02 and the 20 appearance of new peaks at 1.5 (broad s,2) and 3.5 (broad s,4).

Example 23 1, s-la,2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy- 5-methyi-b- [metnyi (2-methylaminoethyr) aminoJ-azirino [2',3':3,4]pyrrolo[1,2-a]indole-4,7-dione carbamate This compound was prepared by the procedure described in Example 1, except that a small amount of 30 solid potassium carbonate was added. From 50 mg of mitomycin A and 25 mg of sym-dimethylethylenedjamine was obtained 28 mg (50% yield) of the desired product having a melting point of 99-101°C (decomposition) and providing the following analysis: NMR (CDCl^, TS) : '6'. values in ppm.

Absence of the 6-methoxy peak at 4.02 and the appearance of new peaks at 1.3 (s,1) , 2.5 (s,6), ana 2.7 (s,4).

Example 24 1,s-la,2,s-8,r-8a,s-8b-Hexahydro-8-(hydroxymethyl) -8a-methoxy- 5-methyl-6-[2-(2-hydroxyethylamino)ethylamino]-azirino [2',3':3,4]pyrrolo[1,2-a]indole-4,7-dione carbamate This compound was prepared by the procedure described in Example 1, except that the solvent was dichlcromethane. From 50 mg of mitomycin A and 18 mg of 2-(2-aminoethylamino)ethanol was obtained 35 mg (58% 15 yield) cf the desired product having a melting point of 115°-118°C (decomposition) and providing the following analysis: NMR (CDCl^r TS): 'o' values in ppm.

Absence of the 6-methoxy peak at 4.02 and the 20 appearance of new peaks at 2.7 (broad s,7) and 3.7 (t,3).

Example 25 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy- 5-methyl-6- [2- (2-hydroxyethoxy) ethylamino] -azi~rincT[21 t 3 ' : 3 ,4] pyrrolo[1,2-a]indole-4,7-dione carbamate This compound was prepared by the procedure described in Example 1, except that the solvent was 30 dichloromethane. From 60 mg of mitomycin A and 20 mg of 2-(2-aminoethoxy) ethanol was obtained 30 mg (.42% yield) of the desired product having a melting point of 99°-102°C (decomposition) and providing the following analysis: NMR (CDCl^, TS) : '5'. values in ppm.

Absence of the 6-methoxy peak at 4.02 and the' appearance of new peaks at 3.3-3.9 (broad s,91 and 6.4-6.8 (broad s,l).

Example 26 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5 -methyl-6-[2-(4-imidazoly1)ethylamino]-azirino[2' , 3 ' ;3 , 4] pyrrolo[1,2-a]indole-4,7-dione carbamate This compound was prepared by the procedure described in Example 1, except that 128 mg of sodium methoxide was added. From 70 mg of mitomycin A and 36 8 mg of histamine dihydrochloride was obtained 61 mg (71% 15 yield) of the desired product having a melting point of 72°-73°C (decomposition) and providing the following analysis: NMR (DMSO-dc, TS): '6' values in ppm. o Absence of the 6-methoxy peak at 4.02 and the appearance of new peaks at 3.0-3.27 (m,4), 7.5 (s,l), 8.0-8.7 (broad s,2) and 8.1 (s,l).

Example 27 1, s-la,2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy- 5-me~thyl-6- [ (2-nitro-l-imidazolyl) ethylamino] -azirino [2 ' , 3':3,4]pyrrolo[1,2-a]indole-4, 7-dione carbamate This compound was prepared by the procedure described in Example 1. From 72 mg of mitomycin A and 30 excess 1-(2-aminoethyl)-2-nitroimidazole was obtained 60 mg (70% yield) of the desired product haying a melting point of 83°-85°C (decomposition) and providing the following analysis: NMR (CDCl^f TS) : '<5' values in ppm. 35 Absence-of the 6-methoxy peak at 4.02 and the appearance of new peaks at 3.4 (t,2), 4.6 (t,2) , 7.3 (broad s,2) and 7.6 (sv 11." Example 28 1, s-la,2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-roethoxy- 5-methy1-6-[2-(4-hydroxyphenyl)ethylamino]-azirino[2',3':3,4] pyrrolo[1,2-a]indole-4,7-dione carbamate This compound was prepared by the procedure 10 described in Example 1. From 130 mg of mitomycin A and 510 mg cf tyramine was obtained 138 mg (81% yield) of the desired product having a melting point of 120°-125°C (decomposition) and providing the following analysis: NMR (CDCl^/ TS): 'c' values in ppm.

Absence of the 6-methoxy peak at 4.02 and the appearance of new peaks at 2.6 (t,2), 2.8 (t,2), 6.7 (a,2), 7.0 (a,2) and 8.0 (s,l).

Example 29 1,s-la,2,s-8,r-8a,s-8b-Hexahydro-8-(hydroxymethyl)-8a-methoxy- 5-"methy1-6-[2-(3,4-dihydroxyphenyl)ethylamino]-azirino [2',3':3,4]pyrrolo[1,2-a]indole-4,7-dione carbamate This compound was prepared by the procedure described in Example 1, except that 138 mg of sodium methoxide was added. From 110 mg of mitomycin A and 660 mg of 3-hydroxytyramine hydrobromiae was obtained 60 mg (40% yield) of the desired product decomposing 30 without melting above 125°C and providing the following analysis: NMR (CDCl^/ TS): '6' values in ppm.

Absence of the 6-methoxy peak at 4.02 and the appearance of new peaks at 2.6 (t,2), 2.8 35 (t,2), 6.4-6.8 (m,3) and 8.3 (broad s,2). ' ' ' - 23 - " Example 30 f- o v . . •• _") 1 rs-la,2,s-8,r-8a,s-8b-Hexahydro-8-(hydroxymethyl) -8a-methoxy- 5-methyl-6-j 2- [ (5-nitro-2-pyridyl) amino] ethylamino J- -azirinc [2',3':3,4]pyrrolo [1,2-a]indole-4,7-dione carbamate This compound was prepared by the procedure described in Example 1, except that the solvent was dichloroir.ethane. From 50 mg of mitomycin A and 30 mg of 2-(2-aminoethylamino)-5-nitropyridine was obtained 40 10 mg (56% yield) of the desired product having a melting point of 76°-79°C (decomposition) and providing the following analysis: NMR (CDCl^/ TS): '6' values in ppm.

Absence of the 6-methoxy peak at 4.02 and the 15 appearance of new peaks at 3.3-4.0 (m,4), 6.2-6.7 (broad s,2), 8.1 (d,l), 8.2 (d,l) and 9.0 (s,1) .

Example 31 1 r s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy- 5 -methyl-6-[2-(1-piperazinyl)etnyiamino]-azirino[2',3':3,4] pyrrolo[1,2-a]indole-4,7-dione carbamate This compound was prepared by the procedure 25 described in Example 1, except that the solvent was dichloromethane. From 50 mg of mitomycin A and 20 mg of N-(2-aminoethyl) piperazine was obtained 23 mg (36% yield) of the desired product having a melting point of 138°-141°C (decomposition) and providing the following 30 analysis: NMR (CDCl^, TS): '6' values in ppm.

Absence of the 6-methoxy peak at 4.02 and the appearance of new peaks at 1.6-2.1 (broad s,l), 2.2-2.6 (broad s,8), 2.6-2.8 (broad 35 s,4) and 6.9 (t,l). - 24" - Example 32 1, s-la f 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy- 5-methvl-6- [ 2- (2-pyridyl). ethylamino J -azirino [2 ' , 3 ' : 3 , 4 j pyrrolo[1,2-a]indole-4,7-dione carbamate This compound was prepared by the procedure described in Example 1. From 70 mg of mitomycin A and 250 mg excess of 2-(2-aminoethyl)pyridine was obtained 51 mg (56% yield) of the desired product having a 10 melting point of 64°-77°C (decomposition) and providing the following analysis: NMR (CDCl^/ TS) : ' <5 1 values in ppm.

Absence of the 6-methoxy peak at 4.02 and the appearance of new peaks at. 2.8 (m,4), 7.0-7.8 15 Cm,3) and 8.5 (d,l).

With specific reference to the compounds comprehended by formula Ilia, the above examples illustrate the following structural variations. 1. Compounds wherein Z is a hydroxy substituted 1-pyrrolidinyl radical represented by Example 1. ' 2. Compounds wherein Z is a lower alkyl substituted piperidyl radical represented by Example 2. 3. Compounds wherein Z is a 1-piperazinyl radical or an acetamino, acetyl, carbamido, cyano, carboxy lower alkylamino, di-lower alkoxy, nitro, sulfamyl or lower alkyl substituted anilino radical are represented, respectively, by Examples 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 and 16. 4. Compounds wherein Z is a radical of the formula R » 1 1 -N-R and wherein R is a nitrogen containing heterocylic radical selected from the group consisting of amino substituted triazolyl, lower alkyl substituted isothiazolyl benzothiazolyl and nitro and halo substituted derivative of benzothiazolyl are represented, respectively, by Examples 17, 18, 19, 20 and 21.

. Compounds wherein Z is a radical of the formula i 1 1 1 .

-N-R and wherein R is a substituted lower alkyl radical selected from the group consisting of amino lower alkyl, lower alkylamino lower alkyl, hydroxy lower alkylamino lower alkyl, hydroxy lower alkoxy lower alkyl, imidazolyl lower alkyl, nitro substituted imidazolyl lower alkyl, mono- and di-hydroxy phenyl lower alkyl, nitro substituted pyridylamino lower alkyl, piperazinyl lower alkyl and pyridyl ethyl are represented, respectively, by Examples 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 and 32.

While none of the foregoing examples are illustrative of compounds wherein Y is other than hydrogen, compounds wherein Y is lower alkyl are nonethe less within the comprehension of the invention, referenc 20 being made to analogously substituted compounds of my aforesaid U.S. Patent Nos. 4, 268, 676 and 4, 460, 599 .. and New Zealand Patent Specification No. 199,617.

Compounds according to the present invention are believed to possess anti-bacterial activity against 25 gram-positive and gram-negative microorganisms in a manner similar to that observed for the naturally occurring mitomycins and are thus potentially useful as therapeutic agents in treating bacterial infections in humans and animals.

Usefulness of compounds of formula Ilia in the antineoplastic therapeutic methods of the invention is demonstrated by the results cf in vivo screening procedures wherein the compounds are administered in varying dosage amounts to mice in which a P38 8 leukemic 35 condition is induced. The procedures were carried out according to "Lymphocytic Leukemia P38 8 - Protocol 1069 1.200", published in Cancer Chemotherapy Reports, Part 3, Vol. 3, No. 2, page 9 (September, ,1972). Briefly put, the screening procedures involved administration of the test compound to CDF^" female mice previously g infected with 10 ascites cells implanted intraperitoneally. Test compounds were administered on the first day of testing only, and the animals were monitored for vitality, inter alia, over a 35-day period.

Results of screening of compounds of Examples 10 1 through 32 are set forth in Table I below. Data given includes optimal dose ("O.D."), i.e., that dosage in mg/kg of body weight of the animal at which the maximum therapeutic effects are consistently observed.

Also included is the median survival time ("MST") 15 expressed as the MST of the test animals compared to the MST of controls x 100 ("% T/C"). Within the context of the in vivo P388 procedure noted above, a % T/C value of 125 or greater indicates significant antineoplastic therapeutic activity. The lowest dose in 20 mg/kg of body weight at which the 125% T/C value is obtained is known as the minimum effective dose ("MED"). These doses also are listed in Table I. It is worthy of note that the exceptionally high MST values obtained in the P388 screenings reported in Table I are also 25 indicative of the absence of substantial toxicity of the compounds at the dosages indicated.

TABLE 1 Example Optimal Dose MST MED No. mg/kg as % T/C 1 25.6 163 0.8 2 25.6 238 <0.2 3 12.8 200 0.2 4 25.6 >333 <0.2 25.6 231 0.2 10 6 6.4 167 0.4 7 25.6 194 1.6 8 3.2 150 0.8 9 12.8 172 <0.2 25.6 322 0.8 15 11 12.8 >333 0.2 12 6.4 161 0.4 13 3.2 172 >0.2 14 25.6 225 0.2 . 12.8 167 0.4 20 16 12.8 181 0.4 17 12.8 181 1.6 18 25.6 169 0.8 19 25.6 150 12.8 25.6 128 25.6 25 21 25.6 144 1.6 22 3.2 178 0.4 23 25.6 133 12.8 24 12.8 133 12.8 25.6 181 0.4 30 26 25.6 163 1.6 27 25.6 150 3.2 28 25.6 218 1.6 29 12.8 139 12.8 12.8 144 6.4 35 31 25.6 138 12.8 32 -25.6 >375 0.2

Claims (6)

2 0 & 9 3 2 - 28 - Clearly among the most preferred compounds employed as antineoplastic agents according to the invention are those exhibiting more than twice the relative life-extending capacity generally characterized 5 as evidencing significant therapeutic potential, i.e., those having an MST % T/C value greater than 2 x 125. The class of such compounds is seen to include the compounds of Examples 4, 10, 11 and 32. As may be noted from Table I, initial single 10 dosages of as little as 0.2 mg/kg showed substantial long term antineoplastic activity. Accordingly, the methods of the invention may involve therapeutic administration of unit dosages of as little as 0.001 mg or as much as 5 mg, preferably from 0.004 mg to 10 mg, of the 15 compounds as the active ingredient in a suitable pharmaceutical preparation. Such preparations may be administered in a daily regimen calling for from 0.1 mg to 100 mg per kg, preferably from about 0.2 to about 51.2 mg per kg, of the body weight of the animal 20 suffering from neoplastic disease. It is preferred that the compounds be administered parenterally. Pharmaceutical compositions suitable for use in practice of methods of the invention may comprise simple water solutions of one or more of the compounds of formula 25 Ilia, but may also include well known pharmaceutically acceptable diluents, adjuvants and/or carriers such as saline suitable for medicinal use. Further aspects and advantages of the present invention are expected to occur to those skilled in the 30 art upon consideration of the foregoing description and consequently only such limitations as appear in the appended claims should be placed thereon. 35 206932 - 29 - yY ] ■ J j\ j / ,Y •*- .0 i— v.: ■ i I • WIIAT IC CLAIMED 13:

1. Compounds of the formula, 5 0 o wherein: Y is hydrogen or lower alkyl; and X is an hydroxy substituted 1-pyrrolidinyl radical, 15 or a lower alkyl substituted 1-piperidyl radical, or an acetamino, acetyl, carbamido, cyano, carboxy lower alkylamino, di-lower alkoxy, nitro, or 20 sulfamyl substituted anilino radical, or R a radical of the formula, -L-R^" wherein R is hydrogen or lower alkyl and R"*" is a nitrogen containing heterocyclic radical selected 25 from the group consisting of amino substituted triazolyl, lower alkyl substituted isothiazolyl, benzothiazolyl, and nitro and halo substituted derivatives of benzothiazolyl, or R^" is a substituted lower alkyl radical selected 30 from the group consisting of amino lower alkyl, lower alkylamino lower alkyl, hydroxy lower alkylamino lower alkyl, hydroxy lower alkoxy lower alkyl, imidazolyl lower alkyl, nitro substituted imidazolyl lower alkyl, mono- and di-hydroxy phenyl lower alkyl, nitro substituted 35 pyridylamino lower alkyl, and piperazinyl lower alk^l. \ 17 NOV 1986n & ...... -30- ' fj \ i . rp.n

2. The .carpaunds according to Claim 1 named: & o , 1,s-la,2,s-8,r-8a,s-8b-Hexahydro-8-(hydroxymethyl) -8a- ' P methoxy-5-methyl-6-(3-hydroxv-l-pyrrolidinyl)-azirino [21,3':3,4]pyrrolo CI,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methaxy-5-fnethyl-6-(3-methylpiperidin-l-yl) -azirino[2' ,3' :3,4] pyrrolo[1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8— (hydroxymethyl) -8a-methoxy-5-methyl-6-[4-(acetylamino)anilino]-azirino 10 [21,3':3,4]pyrrolo(1,2-a]indole-4,7-dicne carbamate; \ 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a- methcxy-5-methvl-6-[3-(acetylamino)anilino]-azirino [21,31:3,4]pyrrolo[1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-15 methoxy-5-methy1-6-(4-acetylanilino)-azirino[21,3':3,4] pyrrolo[1,2-a]indole-4,7-dione carbamate; 1,s-la,2,s-8,r-8a,s-8b-Hexahydro-8-(hydroxymethyl)-8a-methcxy-5-iaethvl-6-[4-(1-ureido)anilino]-azirino[2',3':3,4] pyrrolo(1,2-a)indole-4,7-dione carbamate; 20 ifs-la,2,s-8,r-8a,s-8b-Hexahydro-8-(hydroxymethyl)-Sa me thoxy- 5 -methyl-6- (4-cyanoanilino)-azirino[2',31:3,4] pyrrolo[1,2-a]indole-4,7-dione carbamate; 1/ s-la, 2, s-8, r-8a, s-8b-Hexahydr o-8- (hydr oxymethy 1) -8a- = methoxy-5-methyl-6-(3-cvanoanilino)-azirino[2',31:3,4] 25 pyrrole[1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-5a, s-8b-Hexahydro-8- (hydroxymethyl) -Same thoxy-5-me"thyl-6- [4-(N-glycyl)anilino]-azirino[21,3's3,4] pyrrolo[1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-30 methoxy-5-methyl-6-(3,4-dimethoxyanilino)-azirino[21,3':3,4] pyrrolo[1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-xaethoxy-5-methyl-'6- (3 , 5-dime thoxy anilino) -azirino [2 1 , 3 1 :3, 4] pyrrolo[1,2-a]indole-4,7-dione carbamate; 35 l, s-la, 2, s-8, r-8a,s-8b-Hexahydro-8- (hydroxymethyl) -Sa me thoxy-5-methyl-6-(4-nitroanilino)-azirino[21,3':3,4] pyrrolo[1,2-a]indole-4,7-dione carbamate; l,s-la,2,s-8,r-8a,s-8b-Hexahydro-8-(hydroxyinethyl)-8a-methoxy-5-methyl-6- (4-sulfamylanilino) -azirino [2 ' ,3' :3,4] pyrrclo[1,2-a]indole-4r7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-5 methoxy-5-methyl-6-[(5-amino-l,2,4-triazol-3-yl)amino]- azirino[2', 3 ':3,4]pyrrclo[1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydr o-8- (hydroxymethyl) -Same thoxy-5-methyl-6- [(3-methylisothiazol-5-yl)amino]-azirino [233,4]pyrrolo[1,2-a]indole-4,7-dione carbamate; lu 1' s—laf 2, s—81 r—8a, s—8b—Hexahydro—8— (hydroxymethyl) —8a— methoxy-5-methyl-6-[(2-benzothiazolyl)amino]-azirino [2' , 3':3,4]pyrrole[1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8 ,r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-5-methvl-6-[(6-nitrobenzothiazcl-2-yl)amino]-azirino 15 [231:3,4]pyrrolo[1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxv-5-methyl-6-[(4-chlorobenzothiazol-2-yl)amino]-azirino[2',3':3,4]pyrrolo[1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydr oxyme t hy 1) - 8 a-20 methoxy-5-methyl-6-[(2-aminoethyl)amine]-azirino[2',3',3,4] pyrrolo[1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -Same thoxy-5-me thy 1-6- [methyl (2-methylaminoethyl) amino] -azirino j; [233,4]pyrrolo[1,2-a]indole-4,7-dione carbamate; 25 1, s-la ,2,s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -Sa me thoxy-5-me thy1-6-[2-(2-hydroxyethylamino)ethylamino]-azirino[21,31:3,4]pyrrolo[1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6-[2-(2-hydroxyetnoxy)ethylamino]-azirino .30 [231:3,4]pyrrolo[1,2-a]indole-4,7-dione carbamate; 1, s-la f 2, s-8 ,r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -Same thoxy—5-me thy 1-6- [2-(4-imidazolyl}ethylamino]-azirino [2',3':3,4]pyrrolo[1,2-a]indole-4,7-dione carbamate; 1,s-la,2,s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-35 . methoxy-5-methyl-6-[(2-nitro-l-imidazolyl)ethylamino]- azirino[2',3':3,4]pyrrolo[1,2-a]indole-4,7-dione carbamate; 10 - 32 - cT« LP 1, s-la , 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy~5-methyl-6-[2- (,4-hydroxyphenvl) ethylamino]-azirino .. [2 1 , 3 ':3,4]pyrrolo[1,2-a]indole-4,7-dione carbamate; 1ts-la,2,s-8,r-8a,s-8b-Hexahydro-8-(hydroxymethyl)-Same thoxy- 5 -me thy 1-6- [2-(3,4-aihydroxyphenvl)ethylamino]-azirino[2',3':3,4]pyrrclo[1,2-a]indcle-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6-{2-I(5-nitro-2-pyridyl)amino]ethylamino}-azirino[21,3':3,4]pyrrolo [1,2-a]indole-4,7-dione carbamate;and 1,s-la,2,s-8,r-8a,s-8b-Hexahydro-8-(hydroxymethyl)-8a-methoxy-5-methyl-6-[2-(1-piperazinyl)ethylamino]-azirino [2' , 3':3,4]pyrrolo[1,2-a]indole-4,7-dione carbamate. .0

3. A tnethod for treatment of a neoplastic disease 15 state in a non-human animal, said method comprising administering to said animal having such a disease a therapeutically effective amount of a compound of the formula, 20 25 30 is or or ¥ CH2OCNH2 OCH. Ilia S-la wherein: Y is hydrogen or lower alkyl; and Z an hydroxy substituted 1-pyrrolidinyl radical, a lower alkyl substituted 1-piperidyl radical, a 1-piperazinyl radical or an acetamino, acetyl, carbamido, cyano, carboxy lower alkylamino, di- lower alkoxy, nitro, sulfamyl, or lower alkyl substituted 35 anilino radical, cr - 33 - 10 R 15 a radical of the formula, -N-R wherein R is hydrogen or lower alkyl" and R^" ■ is a nitrogen containing heterocyclic radical selected from the group consisting of amino substituted triazolyl, lower alkyl substituted isothiazolyl, benzothiazolyl, and nitro and halo substituted derivatives of benzothiazolyl, or R^" is a substituted lower alkyl radical selected from the group consisting cf amino lower alkyl, lower alkylamino lower alkyl, hydroxy lower alkylamino lower alkvl, hydroxy lower alkoxy lower alkyl, imidazolyl lower alkyl, nitro substituted imidazolyl lower alkyl, mono- and di-hydroxy phenyl lower alkyl, nitro substituted pyridylamino lower alkyl, piperazinyl lower alkyl, and pyridyl ethyl. -0 % :v

4. The method of Claim 3 wherein the compound is selected from the croup consisting of: 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-20 methoxy-5-methyl-6-(3-hydroxy-l-pyrrolidinyl) -azirino [2 ' , 3 ' : 3 , 4 ]pyrrolo(1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a- methoxy-5-niethyl-6-(3-methylpiperidiii-l-yl)-azirino[2' ,3' ;3,4] pyrrolo[1,2-a]indole-4,7-dione carbamate; 25 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a- methoxy-5-methyl-6-(1-piperazinyl)-azirino[2 1 , 31:3,4] pyrrolo[1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6-[4- (acetylamino)anilino]-azirino .30 [23 3,4 ] pyrrolo (1, 2-a] indole-4 , 7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -Same thoxy-5-me thy 1-6- [3-(acetylamino)anilino]-azirino [2',3':3,4]pyrrolo [1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-35 methoxy-5-methvl-6- (4-acetylanilino)-azirino[21 , 3 ' :3,4] pyrrolo[1,2-a]indole-4,7-dione carbamate; ' v ' ' * 34 <:£?_ 1, s-la,2,s-8 f r-8a,s-8b-Hexahydro-8-(hydroxymethyl)-8a- 0-Pv 3 methoxy-5-methvl-6- 14- (1-ureido) anilino] -a2iri.no [2 ' , 3 ' : 3 ,4] r- pyrrolo(1,2-a]indole-4,7-dione carbamate; 1, s-la ,2, s-8 r r-8a, s-8b-Hexahydro-&~- (hydroxymethyl) -8a- 5 methoxy-5-methyl-6- (4-cyanoanilino)-azirino[21,3':3,4] pyrrolo[1,2-a]indole-4,7-dione carbamate; 1,s-la,2,s-8 , r-8a,s-8b-Hexahydro-8-(hydroxymethyl)-8a-methoxy-5-methyl-6- (3-cyanoanilino)-azirino[2', 31:3,4] pyrrolo[1,2-a]indole-4,7-dione carbamate; 10 1", s-la ,2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a- methoxy-5-methyl-6-[4-(K-glycyl)anilino]-azirino[2',31:3,4] pyrrclo[1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -Same thoxy-5-methyl-6- (3 ,4-dime thoxy anilino) -azirino [2 ' , 3 1 :.3 , 4] 15 pyrrole[1,2-a]indole-4,7-dione carbamate; 1,s-la,2,s-8,r-8a,s-8b-Hexahydro-8-(hydroxymethyl)-Same thoxy -5 -me thy 1-6- (3,5-dimethoxyanilino)-azirino[2',3':3,4] pyrrolo[1,2-ajindole-4,7-dione carbamate; 1,s-la,2,s-8,r-8a,s-8b-Hexahydro-8-(hydroxymethyl)-8a- 20 methoxy-5-methyl-6 - (4-nitroanilino)-azirino[2',3':3,4] pyrrolo[1,2-a]indole-4,7-dione 'carbamate; 1,s-la,2,s-8,r-8a,s-8b-Hexahydro-8-(hydroxymethyl)-8a-methoxy-5-methyl-6- (4-sulfamylanilino)-azirina[2' , 3 1 : 3 ,4 ] pyrrolo[1,2-a]indole-4,7-dione carbamate; 1,s-la,2,s-8, r-8a, s-'8b-Hexahydr 0-8- (hydroxymethyl) -Same thoxy-5-methyl-6- (4-methylanilir>o) -azirino [2 ' , 31:3,4] pyrrolo[1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -Same thoxy-5-methyl-6- (3-methylanilinc)-azirino[21,31;3,4 3 30 pyrrole[1,2-a1indole-4,7-dione carbamate; 1,s-la,2,s-8,r-8a,s-8b-Hexahydro~8-(hydroxymethyl)-Same thoxy -5 -me thy 1-6- [(5-amino-l,2,4-triazol-3-yl)amino]-azirino[2',3':3,4]pyrrolo[1,2-a]indole-4,7-dione carbamate; 1,s-la,2,s-8,r-8a,s-8b-Hexahydro-8-(hydroxymethyl)-8a-35 methoxy-5-methyl-6-i (3-methvlisothiazol-5-yi)amino]-azirino [2',3':3,4]pyrrolo [1,2-a]indole-4,7-dione carbamate; - 35 - " ^ ^ ■ '~^9 1, s-la, 2, S78, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) - ''cr ~ ^ 8a-methoxy-5-methyl-6- [ (2-benzothiazolyl)amino]-azirino ^3 [2',3' :3,4]pyrrolo [ 1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8,r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a- 5 methoxy-5-methyl-6-[ (6-nitrobenzothiazol-2-yl)amino]-azirino [2',3':3,4]pyrrolo [1, 2-a]indole-4,7-dione carbamate; 1, s-la ,2,s-8,r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6- [ (4-chlorobenzothiazol-2-yl)amino]-azirino[2',3':3,4]pyrrolo[1,2-a]inacle-4,7-diorie carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -Same thoxy-5-methyl-6- [(2-aminoethvl)amino]-azirino[21,3':3,4] pyrrolo[1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a- methoxy-5-methvl-6-[methyl(2-methylaminoethyl)amino]-azirino 15 [21,33,4]pyrrolo[1,2-a]indole-4,7-dione carbamate; 1, s-la , 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -Same thoxy-5-me thy 1-6- [2- (2-hydroxyethylamino)ethylaminoj-azirino [ 2 ' , 3 1 :3,4]pyrrclo[1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a,s-8b-Hexahydro-8- (hydroxymethyl) -8a-20 methoxy-5-methyl-6-[2-(2-hvdroxyethoxy)ethylamino]-azirino [21,31:3,4]pyrrolo[1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6-[2-(4-imidazolvl) ethylamino.] -azirino [21,3':3,4]pyrrolo[1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a- methoxy-5-methvl-6- [ (2-nitro-l-imidazolyl)ethylaminoj-azirino[2',31:3,4]pyrrolo[1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -Same thoxy-5-me thy 1-6- [2- (4-hydroxyphenyl)ethylamino]-azirino 30 [231:3,4]pyrrolo[1,2-a]indole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-Sb-Hexahydro^S- (hydroxymethyl) -Same thoxy-5-me thyl-6-[2- (3,4-dihydroxypnenyl)ethylamino]-azirino[2',3':3,4]pyrrolo[1,2-alindole-4,7-dione carbamate; 1, s-la, 2, s-8, r-8a, s-8b-Hexahydr 0-8- (hydroxymethyl) -8a-35 methoxy-5-methyl-6-{2-[(5-nitro-2-pyridyl)amino]ethylamino;- I^^ ■" ''J I > - 36 - 1, s-la, 2 , s-8, r-8a,s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6-[2-(1-piperazinyl)ethylamino]-azirino . [2',3' : 3,4]pyrrolo[1,2-a]indole-4,7-dione carbamate; and • ■* 1, s-la, 2, s-8, r-8a, s-8b-Hexahydro-8- (hydroxymethyl) -8a-methoxy-5-methyl-6-[2- (2-pyridyl)ethylamino]-azirino [2',3':3,4]pyrrolo[1,2-a]indole-4, 7—dione carbamate. O o. C . o 10

5. The method of claim 3 wherein the-amount of the compjund administered comprises a daily dose of from __ -0.2 mg to 51.2 mg per kg of the body weight of the animal.

6. A pharmaceutical composition for use in treatment of a neoplastic disease in an animal, said 15 composition comprising a pharmaceuticallv acceptable solvent, diluent, adjuvant or carrier and, as the active ingredient, from .. 0.001 to ' 5 mg of a compound of the formula O 20 OCH- 25 ^ch2ocnh2 S -I£ is 30 or or wherein: Y is hydrogen or lower alkyl; and Z an hydroxy substituted 1-pyrrolidinyl radical, a lower alkvl substituted 1-piperidyl radical, a 1-piperazinyl radical or an acetamino, acetyl, carbamido, cyano, carboxy lower alkylamino, di- 35 lower alkoxy, nitro, sulfamyl, or lower alkyl substituted anilino radical, or - 37 - R a radical of the formula, -N-R^" wherein R is hydrogen or lower alkyl and R"1" is a nitrogen containing heterocyclic radical selected from the group consisting of amino substituted tri-5 azolyl, lower alkyl substituted isothiazolyl, benzothiazolyl, and nitro, and halo substituted derivatives of benzo- from the group consisting of amino lower alkyl, lower 10 alkylamino lower alkyl, hydroxy lower alkylamino lower alkyl, hydroxy lower alkoxy lower alkyl, imidazolyl lower alkyl, nitro substituted imidazolyl lower alkyl, mono- and di-hydroxy phenyl lower alkyl, nitro substituted pyridylamino lower alkyl, piperazinyl lower 15 alkyl, and pyridyl ethyl. thiazolyl, or R^" is a substituted lower alkyl radical selected U&m&dty. ByJ4+6/Their Authorised .Agerrts, A. J. PARK & SON 20 25 30 35