FI80698C - FOERFARANDE FOER FRAMSTAELLNING AV TERAPEUTISKT ANVAENDBARA DERIVAT AV MITOMYCIN A. - Google Patents

Description

1 80698

This invention relates to a process for the preparation of novel therapeutically useful mitomycin A derivatives. The compounds are used in the treatment of tumor conditions in animals.

U.S. Patent Nos. 4,268,676, 4,460,599 and 4,617,389 to the Applicant are incorporated herein by reference in their entirety to the extent that they are capable of providing essential or less important material in connection with the present invention.

In short, the aforementioned U.S. Patent Nos. 4,268,676 and 4,460,599 provide background information for the ongoing 15 research aimed at new and useful

to compounds which are structurally related to mitomycin, have antibiotic activity and low toxicity, as well as significant antitumor activity in animals. More specifically, they describe 20 new compounds of formula I.

0?

X "A / CH.OCNH 2 I

Wherein Y is hydrogen or lower alkyl; and X is a thiazole amino radical, a furfural amino radical or a radical of formula R R 35 -NC-R 1 R 2 80698 wherein R, R 1 and R 2 are the same or different groups which are hydrogen and lower alkyl, and R 3 is lower alkenyl, lower haloalkenyl, lower alkynyl, lower alkoxycarbonyl, thienyl, formamyl, tetrahydrofuryl or Benz-5-sulfonamide. The aforementioned U.S. patents also disclose novel methods for treating tumor disease states in animals comprising administering a therapeutically effective amount of a compound of formula Ia 10 o 0 Ζ'Ύ \ _ / CH2 ° "™ 2

'3C

0 0 '- wherein Y is hydrogen or lower alkyl; and Z is a thiazole amino radical, a furfurylamino radical, a cyclopropylamino radical, a pyridylamino radical or a radical of the formula R 4 R 5 I - N - C - R 7 if 25 wherein R 4, R 5 and R 6 are the same or different groups which are hydrogen or lower alkyl, and R 7 is lower alkenyl, lower haloalkenyl, lower alkynyl, lower alkoxycarbonyl, lower haloalkyl, lower hydroxyalkyl, pyridyl, thienyl, formamyl, tetrahydrofuryl, benzyl or benzenesulfonamide.

U.S. Patent No. 4,627,389 also discloses compounds having significant antitumor activity in animals and having the formula Ha

II

Where Y is hydrogen or lower alkyl; and Z is a (lower alkoxy) -substituted quinolinylamino radical, a cyano-substituted pyrazolylamino radical or a mono- or disubstituted mono- or disubstituted by a lower alkyl. a nitrogen-containing heterocyclic radical which is 1-pyrro-linyl, 1-indolinyl, N-thiazolidinyl, N-morpholinyl, 1-piperazinyl or N-thiomorpholinyl, or cyano, phenyl, carboxamido or (lower alkoxycarbonyl) a substituted 1-aziridinyl radical, or a (lower alkyl), formyl or acetylphenyl-substituted 1-piperazinyl-20-liradical radical, or a hydroxy- or piperidyl-substituted 1-piperidyl radical, or a (lower alkoxy), amino- or halo-substituted amino radical , a mercapto- or methylenedioxy-substituted aniline radical, or a radical of formula 25

R

-NR 'wherein R is hydrogen or lower alkyl and R' is a nitrogen-containing heterocyclic radical selected from quinuclidinyl, pyrazolyl, 1-triazolyl, isoquinolinyl, indazolyl, benzoxazolyl, thiadiazolyl or benzothiadiazolyl or lower alkyl or lower alkyl thereof. a halogen-substituted derivative, or a butyrolactonyl radical, 35 or an adamantyl radical; or a lower alkoxy monosubstituted phenyl radical, or a substituted lower 4,80698 alkyl radical which is lower mercaptoalkyl, lower carboxyalkyl, mono-, di- or tri (lower alkoxy) (lower alkyl), (lower alkyl) thio (lower alkyl) or their lower alkoxycarbonyl-substituted derivative, lower cyanoalkyl, mono-, di- or tri (lower alkoxy) phenyl (lower alkyl), (lower N-alkyl) pyrrolidinyl (lower alkyl), N-morpholinyl (lower alkyl) or (lower dialkylamino) (lower alkyl).

Also related to the background of the present invention are the following references:

Coluslich et al., U.S. Patent 3,332,944;

Matsui et al., U.S. Patent 3,410,867;

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

Matsui et al., U.S. Patent 3,429,894; 15 Remers, U.S. Patent 4,268,676;

Matsui et al., U.S. Patent 3,514,452; and Imai et al., Gann. 71 (1980), ss. 560 - 562.

This invention relates to a process for the preparation of therapeutically useful mitomycin A derivatives of formula III

0? X JI / ch2ocnh2 I I L ^ OCH, I L_J> 1 2 3 4 5 6 li wherein Y is hydrogen or lower alkyl; and X is a hydroxy-substituted 1-pyrrolidinyl radical or a lower alkyl-substituted piperidinyl radical or an acetamido, 4-acetyl, urea, cyano, carboxy (lower alkyl-amino) -, di- (lower alkoxy) -, nitro- or sulfamyl-substituted aniline radical or radical of formula 5 80698

R

-N-R 1 wherein R b is hydrogen or lower alkyl and R 1 is a nitrogen-containing heterocyclic radical which is amino-substituted triazolyl, lower alkyl-substituted isothiazolyl, benzothiazolyl or a nitro- or halogen-substituted alkyl derivative of benzothiazolyl, or which is hydroxy (lower alkylamino) (lower alkyl), hydroxy (lower alkoxy) (lower alkyl), imidazolyl (lower alkyl), (nitro-substituted imidazolyl) (lower alkyl), (nitro-substituted pyridylamino) (lower alkyl) or piperazinyl (lower alkyl).

Unless otherwise indicated, the term "lower" when applied to "alkyl" radicals means straight-chain or branched radicals containing 1 to 6 carbon atoms. "Lower alkyl" is, for example, methyl, ethyl, propyl, butyl, pentyl or hexyl, as well as isopropyl, t-butyl or the like. Likewise, the term "lower" in the context of "alkoxy" means a radical having 1 to 6 carbon atoms.

All of the novel antibiotic mitomycin derivatives of formula III are useful in the application of new antitumor therapies.

The process for the preparation of mitomycin derivatives according to the invention is characterized in that mitomycin A or N-alkyl-mitomycin A is reacted with an amine which is pyrrolidinone, or a lower alkylpiperidine, or acetamino, acetyl, uramido, cyano, carboxy (lower alkylamino) - di (lower alkoxy) -, nitro- or sulfamyl-substituted aniline, or 35 amino or a (lower alkyl) substituted derivative thereof which is amino-substituted triazolylamine, 6,80698 (lower alkyl) substituted isothiazolylamine, benzo-thiazine - or halogen-substituted benzothiazolylamine, hydroxy (lower alkylamino) (lower alkylamine), hydroxy (lower alkoxy) (lower alkylamine), imidazolyl (lower alkylamine), nitro-substituted imidazolyl (lower alkylamine), lower alkylamino-substituted (nitroamino), nitro-substituted piperazinyl (lower alkylamine), and the resulting product is isolated from the reaction mixture.

N-alkylmithomycin A is prepared from mitomycin C, for example, by the method described by Cheng et al. have described in general [J. Med. Chem. 20, No. 6 (1977), p. 767-770]. The production reactions generally give the desired product as a solid which is highly soluble in alcohol. The compounds of this invention are believed to have antibacterial activity against gram-positive and gram-negative microorganisms in the same manner as naturally occurring mitomycins, and thus are potentially useful as therapeutic agents for the treatment of bacterial infections in humans and animals.

The utility of the compounds of the invention in methods of treating tumors is demonstrated by the results obtained in in vivo experiments in which the compounds are administered at varying doses to mice induced with P388 leukemia. The experiments were performed according to Lymphocytic Leukemia P388 - Protocol 1,200 [Cancer Chemotherapy Reports, Part 3, Door.3, No. 2 (September 1972) p. 9]. Briefly, in experiments, 30 test compounds were administered to female CDF1 mice pre-infected by implantation of 106 peritoneal cells intraperitoneally. The test compounds were administered only on the first day of the experiment, and the animals were monitored e.g. their vitality over 35 days.

Il 7 80698

The experimental results obtained for the compounds of Examples 1-27 are shown in Table I. The data given include the optimal dose ("O.D."), i. the dose (mg per kg body weight of the animal) at which the best Hoi-5 effects are achieved permanently. The table also gives the mean survival time ("MST") expressed as (MST of test animals) / (MST of control animals) x 100 ("% T / C"). In connection with the above in vivo P388 procedure, a% T / C of 125 or greater indicates a significant anti-tumor therapeutic effect. The lowest dose (mg / kg body weight) at which a% T / C value of 125 is obtained is known as the lowest effective dose ("MED"). These doses are also given in Table I. It is noteworthy that the exceptionally high MST values obtained in the P388 experiments, the results of which are shown in Table I, also indicate that the compounds are not significantly toxic at the doses used.

Table I

8 80698

Eg Dose MST MED

n: mg / kg% T / C_ 5 1 25.6 163 0.8 2 25.6 238 <0.2 3 25.6> 333 <0.2 4 25.6 231 0.2 5 6.4 167 0.4 10 6 25.6 194 1.6 7 3.2 150 0.8 8 12.8 172 <0.2 9 25.6 322 0.8 10 12.8> 333 0.2 15 11 6, 4,161 0.4 12 3.2,172> 0.2 13 25.6 225 0.2 14 12.8 167 0.4 15 12.8 181 0.4 20 16 12.8 181 1.6 17 25, 6,169 0.8 18 25.6 150 12.8 19 25.6 128 25.6 20 25.6 144 1.6 25 21 12.8 133 12.8 22 25.6 181 0.4 23 25.6 163 1.6 24 25.6 150 3.2 25 12.8 144 6.4 30 26 25.6 138 12.8 27 25.6> 375 0.2 9 80698

By far the most preferred compounds for use as antitumor agents according to the invention are those which provide a prolongation of life of 2 times that which is considered to be an indication of a significant therapeutic effect, i. compounds with an MST% T / C greater than 2 x 125. The compounds of Examples 3, 9, 10 and 27 are considered to belong to this group.

As shown in Table I, even initial doses as low as 0.2 mg / kg had a significant long-term antitumor effect. Thus, the active compounds can be administered in unit doses of 0.001 to 5 mg, preferably 0.004 to 10 mg, as suitable pharmaceutical preparations. Such preparations may be administered daily at 0.1 to 100 mg, preferably from about 0.2 to 51.2 mg per kilogram of body weight to an animal suffering from a tumor disease. It is preferred to administer the compounds parenterally. Suitable pharmaceutical compositions may also contain known pharmaceutically acceptable diluents, excipients and / or carriers, such as saline suitable for medical use.

The compounds of the invention show that the former are less toxic compared to the compounds of U.S. Patent No. 4,231,936. Table I of the present application provides optimal doses of antitumor therapy for the compounds of this invention. These optimal doses to achieve optimal survival are close to or greater than the LD50-30 values reported in the U.S. patent. The LD50 value is thus the dose that causes death in 50% of animals. This demonstrates the advantages of the present compounds, i.e., that they are certainly less toxic.

Mention should also be made of Examples 35 and III of U.S. Pat. No. 35, i.e. Tables 3 and 4, which show the antitumor effects in the tested animals compared to the control animals as% ILS (% prolongation of life). These values can be compared to the mean survival time (MST) reported in Table I of the present application, expressed as the MST of the test animals compared to 5 control animals x 100 (% T / C). The compounds of the present invention have exceptionally high MST values, which also indicates that the compounds are relatively non-toxic compared to the compounds of U.S. Patent No. 4,231,936.

Other advantages of the present invention will become apparent from the following Examples 1-27, which illustrate the preparation of certain preferred compounds.

Unless otherwise stated, all reactions were performed at room temperature (20 ° C) without heating. Unless otherwise noted, all thin layer chromatography (TLC) experiments used to monitor the progress of the reaction used pre-coated silica gel plates and a 2: 8 v / v mixture of methanol and chloroform as eluent.

Example 1 1,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- (3-hydroxy-1-pyrrolidinyl) -azlorine [2 ', 3': 3 .4] pyrrolo [1,2-a] indole-4,7-dione carbamate A solution of 50 mg of mitomycin A in anhydrous methanol (6 ml) was treated with 3-pyrrolidinol (13 mg) under nitrogen at room temperature. When thin layer chromatography on silica gel plates (eluent: methanol / chloroform 2: 8) showed no more starting material, the mixture was filtered and evaporated under reduced pressure. The residue was purified by preparative thin layer chromatography using the same solvent mixture. There was thus obtained 23 mg (yield 40%) of the desired product, m.p. was 82-85 ° C (decomposed), and analytical data 35 were as follows: il 11 80698 NMR (DMSO-d 6, TS): δ values (ppm): 6-methoxy peak 4.02 is absent and the spectrum has new peaks 1.6 - 2.2 (m, 2), 2.8-3.1 (broad s, 5) and 4.0-4.3 (m, 1).

Example 2 5,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- (3-methylpiperidinyl) -azirine [2 ', 3': 3,4] pyrrole [1,2-a] Indole-4,7-dlonicarbamate This compound was prepared by the procedure of Example 1. Mitomycin A (70 mg) and 3-methylpiperidine (200 mg) gave 46 mg (55% yield) of the desired product, m.p. was 75-88 ° C (decomposed), and the analytical data are as follows: NMR (CDCl 3, TS): δ values (ppm): 6-methoxy peak 4.02 missing and the spectrum has new peaks 0.85 (d, 3 ), 1.10 to 2.15 (m, 5) and 2.15 to 3.32 (m, 4).

Example 3 1,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- [4- (acetylamino) aniline] -20 azirine ^ ', 3': 3.4 ] pyrrolo [1,2-a] indole-4,7-dione carbamate This compound was prepared by the procedure described in Example 1. From mitomycin A (100 mg) and excess 4- (acetylamino) aniline, 102 mg (76% yield) of the desired product were obtained, m.p. was 143-145 ° C (decomposed), and the analytical data are as follows: NMR (CDCl 3, TS): δ values (ppm): 6-methoxy peak 4.02 is absent and the spectrum has new peaks 2.1 (s, 3), 7 , 4 (d, 2), 7.6 (s, 1) and 8.9-9.3 (s, 1).

Example 4 1,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- [3- (acetylamino) aniline] -azine [2 ', 3 *: 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate 35 This compound was otherwise prepared as in Example 1, but a small amount of solid 12,80698 potassium carbonate was added. Mitomycin A (70 mg) and 3- (acetylamino) aniline (150 mg) gave 67 mg (72% yield) of the desired product, m.p. was 140-143 ° C (decomposes) and the analytical data are as follows: δ NMR: (acetone-d 6, TS): δ values (ppm): 6-methoxy peak 4.02 is absent and the spectrum has new peaks 2.1 (s, 3), 6.7-7.5 (m, 4), 8.0 (broad s, 1) and 9.3 (s, 1).

Example 5 10 11a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- (4-acetylaniline) -azine [21,31: 3,4] pyrrole [1,2-a] Indole-4,7-dione carbamate This compound was prepared according to Example 1, except that a small amount of solid potassium carbonate was added. Mitomycin A (70 mg) and 4-acetylaniline (50 mg) gave 25 mg (28% yield) of the desired product, m.p. was 103-104 ° C (decomposed), and the analytical data are as follows: 20 NMR (CDCl 3, TS): δ values (ppm): 6-methoxy peak 4.02 absent and new peaks in the spectrum 2.1 (s, 3) , 6.6 (d, 2), 7.3 (d, 2) and 7.0-7.3 (broad s, 1).

Example 6 1,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-25-methoxy-5-methyl-6- [4- (1-ureido) aniline] -azlorin 82 *, 3 *: 3,4] Pyrrolo [1,2-a] indole-4,7-dione carbamate This urea-substituted compound was prepared according to Example 1. Mitomycin A (50 mg) and 4- (1-30 ureido) aniline (227 mg) gave 49 mg (67% yield) of the desired product, mp 93-95 ° C (decomposed), and the analytical data were as follows: NMR ( CDCl 3, TS): δ values (ppm): 6-methoxy peak 4.02 is absent and the spectrum has new peaks 5.03 (s, 2), 6.9 (d, 2), 35 8.0 (s, 1 ) and 8.4 (s, 1).

Il 13 80698

Example 7 1,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- (4-cyanoaniline) -azirine- [2 ', 3': 3,4] pyrrolo [1,2-a] Indole-4,7-dione carbamate 5 This compound was otherwise prepared according to Example 1, but a small amount of solid potassium carbonate was added. Mitomycin A (70 mg) and 4-aminobenzonitrile (472 mg) gave 23 mg (24% yield) of the desired product, m.p. was 124-126 ° C (decomposed), and the analytical data were as follows: NMR (CDCl 3, TS): δ values (ppm): 6-methoxy peak 4.02 missing and new peaks in the spectrum 6.6 (d, 2) , 7.4 (d, 2) and 7.0-7.3 (broad s, 1).

Example 8 1,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- (3-cyanoaniline) -azirine- [2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate This compound was prepared according to Example 1, but a small amount of solid potassium carbonate was added. Mitomycin A (71 mg) and 3-aminobenzonitrile (500 mg) gave 30 mg (34% yield) of the desired product, m.p. was 97-98 ° C (decomposed), and the analytical data are as follows: 25 NMR (CDCl 3, TS): δ value (ppm): 6-methoxy peak 4.02 is absent and the spectrum has new peaks 7.2-7.8 (m , 4).

Example 9 1,1a, 2,8,8a, 8b-Hexahydro-8-hydrocymethyl-8a-methoxy-5-methyl-6- [4- (N-glycyl) aniline] -30 azirine ^ ', 3': 3 , 4] pyrrolo [1,2-a] indole-4,7-dione carbamate This compound was prepared according to Example 1. Mitomycin A (50 mg) and 4- (N-glycyl) aniline (249 mg) gave 62 mg (90% yield) of the desired product, m.p. was 83-85 ° C (decomposed), and the analytical data are as follows: ---- 14 -___ i4 80698 NMR (DMSO-d6, TS): δ values (ppm): 6-methoxy peak absent and new peaks in the spectrum 3, 1 (s, 2), 6.3 to 6.6 (broad s, 2), 6.6 to 6.8 (broad s, 2), and 6.6 to 7.1 (broad s, 2). Example 10 5,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- (3,4-dimethoxyanilinyl) -azine [21,3 ': 3, 4] Pyrrolo [1,2-a] indole-4,7-dione carbamate This compound was prepared according to Example 1. Mitomycin A (50 mg) and 3,4-dimethoxyaniline (229 mg) gave 61 mg (91% yield) of the desired product, m.p. was 114-116 ° C (decomposed), and the analytical data are as follows: NMR (CDCl 3, TS): δ values (ppm): 6-methoxy peak 4.02 is absent and the spectrum has new peaks 3.8 (s, 6), 6 , 3-6.9 (m, 3) and 7.7 (s, 1).

Example 11 1,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- (3,5-dimethoxyaniline) -azirine [2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-20 dione carbamate This compound was prepared according to Example 1. Mitomycin A (50 mg) and 3,5-dimethoxyaniline (229 mg) gave 60 mg (88% yield) of the desired product, m.p. was 98-100 ° C (decomposed), and the analytical data are as follows: 25 NMR (CDCl 3, TS): δ values (ppm): the 6-methoxy peak 4.02 is absent and the spectrum has new peaks 3.8 (s, 6), 5.9 to 6.4 (broad s, 3) and 7.6 (s, 1).

Example 12 1,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-30-methoxy-5-methyl-6- (4-nitroaniline) -azirine- [2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dionicarbamate This compound was otherwise prepared according to Example 1, but a small amount of solid potassium carbonate was added. Mitomycin A (70 mg) and 4-nitroaniline (276 mg) gave 16 mg (9% yield) of the desired product, m.p. 80698, m.p. was 132-134 ° C (decomposed), and the analytical data are as follows: NMR (acetone-d 6, TS): δ values (ppm): 6-methoxy peak 4.02 is absent and the spectrum has new peaks 6.9-7, 3 (d, 2), δ 7.4-7.9 (d, 2) and 7.9-8.4 (broad s, 1).

Example 13 1,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- (4-sulfamylaniline) -azirine- [2 ', 3 *: 3.4 ] pyrrolo [1,2-a] indole-4,7-dione-10 carbamate This compound was otherwise prepared according to Example 1, but a small amount of solid potassium carbonate was added. Mitomycin A (70 mg) and sulfanilamide (688 mg) gave 25 mg (26% yield) of the desired product, m.p. was 113-115 ° C (decomposed), and the analytical data are as follows: NMR (DCCl 3, TS): δ values (ppm): 6-methoxy peak 4.02 is absent and the spectrum has new peaks 7.0 (d, 2) , 7.5 (s, 1) and 7.9 (d, 2).

Example 14 1,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- (4-methylaniline) -azirine [2 ', 3': 3.4 ] pyrrolo [1,2-a] indole-4,7-dione carbamate This compound was prepared as described in Example 1. Mitomycin A (60 mg) and excess 4-methylaniline gave 63 mg (86% yield) of the desired product, m.p. was 113-115 ° C (decomposed), and the analytical data are as follows: 30 NMR (CDCl 3, TS): δ values (ppm): 6-methoxy peak 4.02 is absent and the spectrum has new peaks 2.3 (s, 3), 6.5 to 7.3 (broad s, 4) and 7.6 (broad s, 1).

35 16 80698

Example 15 1,13,2,8,83,8b-Hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- (3-methyllanillin) -azirine [2 ', 31: 3,4] pyrrolo [1,2-a] Indole-4,7-dione-5-carbamate This compound was prepared according to Example 1. Mitomycin A (70 mg) and 3-methylaniline (276 mg) gave 66 mg (78% yield) of the desired product, m.p. was 89-91 ° C (decomposed), and the analytical data are as follows: 10 NMR (CDCl 3, TS): δ values (ppm): 6-methoxy peak 4.02 absent and new peaks in the spectrum 2.4 (s, 3) , 6.7-7.5 (m, 4) and 7.8 (s, 1).

Example 16 1,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-15-methoxy-5-methyl-6 - [(5-amino-1,2,4-triazol-3-yl) amino] ] -azirine [21,31: 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate This compound was otherwise prepared according to Example 1, but a small amount of solid potassium carbonate was added. Mitomycin A (50 mg) and 3,5-diamino-1,2,4-triazole (30 mg) gave 13 mg (5.5% yield) of the desired product, m.p. was 117-120 ° C (decomposed), and the analytical data were as follows: NMR (DMSO-d 6, TS): δ values (ppm): 6-methoxy peak 4.02 25 is missing and the spectrum has a new peak 5.37 (s , 3). Example 17 1,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-Same-toxo-5-methyl-6 - [(3-methyl-isothiazol-5-yl) -amino] -azirine [2t, 3 ': 3,4] pyrrolo [1,2-a] -30 indole-4,7-dione carbamate This compound was otherwise prepared according to Example 1, but 0.5 ml of triethylamine was added. Mitomycin A. (60 mg) and 5-amino-3-methylisothiazole hydrochloride (30 mg) gave 4.5 mg (8.5% yield) of the desired product, m.p. was 87-90 ° C (decomposed), and the analytical data are as follows: 11 17 80698 NMR (CDCl 3, TS) δ values (ppm): 6-methoxy peak 4.02 absent and new peaks in the spectrum 2.3 (s, 3) , 6.1 (s, 1) and 6.4 (s, 1).

Example 18 5,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6 - [(2-benzothlazolyl) amino] azirine [2 ', 3': 3, 4] pyrrolo [1,2-a] indole-4-dione carbamate This compound was otherwise prepared according to Example 1, but a small amount of solid potassium carbonate was added. Mitomycin A (50 mg) and 2-aminobenzothiazole-Sol (25 mg) gave 12 mg (18% yield) of the desired product, m.p. was 82-85 ° C (decomposed), and the analytical data are as follows: 15 NMR (CDCl 3, TS): δ values (ppm): 6-methoxy peak 4.02 is absent and the spectrum has a new peak 7.1-8.0 (m , 5). Example 19 1,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6 - [(6-nitrobenzothiazol-2-20yl) amino] azirine [2 ', 3 ': 3,4] Pyrrolo [1,2-a] indole-4,7-dione carbamate This compound was otherwise prepared as described in Example 1, but a small amount of solid potassium carbonate was added. Mitomycin A (50 mg) and 2-amino-25-nitrobenzothiazole (30 mg) gave 20 mg (27% yield) of the desired product, m.p. was 86-89 ° C (decomposed), and the analytical data are as follows: NMR (DMSO-d 6, TS): δ values (ppm): 6-methoxy peak 4.02 is absent and the spectrum has new peaks 6.9-8.3 ( m, 4).

Example 20 1,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6 - [(4-chlorobenzothiazol-2-yl) amino] azirine [2,3,3 *: 3,4] Pyrrolo [1,2-a] indole-4,7-dione carbamate This compound was otherwise prepared according to Example 1, but a small amount of solid potassium hard 80698 carbonate was added. Mitomycin A (150 mg) and 2-aminochlorobenzothiazole (27 mg) gave 30 mg of the desired product (yield 14%), m.p. was 89-91 ° C (decomposed), and the analytical data are as follows: δ NMR (CDCl 3, TS): δ values (ppm): 6-methoxy peak 4.02 absent and a new peak in the spectrum 7.1 - 8.0 ( broad s, 4). Example 21 1,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- [2- (2-hydroxyethylamino) ethylamino] azine [2,3,3 ': 3,4] Pyrrolo [1,2-a] indole-4,7-dione carbamate This compound was otherwise prepared according to Example 1, but using dichloromethane as the solvent. From mitomycin A (50 mg) and 2-82-aminoethylamino) ethan-15 (18 mg), 35 mg (58% yield) of the desired product were obtained, m.p. was 115-118 ° C (decomposed), and the analytical data are as follows: NMR (CDCl 3, TS): δ values (ppm): 6-methoxy peak 4.02 is absent and the spectrum has new peaks 2.7 (broad s, 7) and 3.7 20 (t, 3).

Example 22 1,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- [2- (4-imidazolyl) ethylamino] azirine [2,3-g] .4] pyrrolo [1,2-a] indole-25 4,7-dione carbamate This compound was otherwise prepared according to Example 1, but dichloromethane was used as the solvent. Mitomycin A (60 mg) and 2-82-aminoethoxy) ethanol (20 mg) gave 30 mg (42% yield) of the desired product, m.p. was 99-102 ° C (decomposed), and the analytical data are as follows: NMR (CDCl 3, TS): δ values (ppm): 6-methoxy peak 4.02 is absent and the spectrum has new peaks 3.3-3.9 (broad s , 9) and 6.4 to 6.8 (broad s, 1).

35 19 80698

Example 23 1,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- [2- (4-imidazolyl) ethyl-βΜίηο] -βΐ8ίΓϋηί [2 *, 3 ' : 3,4] Pyrrolo [1,2-a] -5-indole-4,7-dione carbamate This compound was otherwise prepared as described in Example 1, but 128 g of sodium methoxide was added. Mitomycin A (70 mg) and histamine hydrochloride (368 mg) gave 61 mg (71% yield) of the desired product, m.p. was 72-73 ° C (decomposed), and the analytical data were as follows: NMR (DMSO-d 6, TS): δ values (ppm): 6-methoxy peak 4.02 missing and new peaks in the spectrum 3.0-3.27 (m, 4), 7.5 (s, 1), 8.0-8.7 (broad s, 2) and 8.1 (s, 1).

Example 24 1,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6 - [(2-nitro-1-imidazolyl) ethylamino] azirine [21.3 ': 3,4] Pyrrolo [1,2-a] indole-4,7-dione carbamate This compound was prepared according to Example 1.

Mitomycin A (72 mg) and excess 1- (2-aminoethyl) -2-nitroimidazole gave 60 mg (70% yield) of the desired product, m.p. was 83-85 ° C (decomposed), and the analytical data are as follows: 25 NMR (CDCl 3, TS): δ values (ppm): 6-methoxy peak 4.02 absent and new peaks in the spectrum 3.4 (t, 2) , 4.6 (t, 2), 7.3 (broad s, 2) and 7.6 (s, 1).

Example 25 1,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-30-methoxy-5-methyl-6- {2- [5-nitro- (2-pyridyl) amino] ethylamino} -azirine [2 *, 3 *: 3,4] -pyrrolo [1,2-a] indole-4,7-dione carbamate This compound was otherwise prepared as described in Example 1, but using dichloro-35 methane as the solvent. Mitomycin A (50 mg) and 2- (2-aminoethylamino) -5-nitropyridine (30 mg) gave 40 mg (yield 20 8 0 6 9 8 56%) of the desired product, m.p. was 76-79 ° C (decomposed), and the analytical data are as follows: NMR (CDCl 3, TS): δ values (ppm): 6-methoxy peak 4.02 is absent and the spectrum has new peaks 3.3-3.0 (m, 4), 6.2 -5.67 (broad s, 2), 8.1 (d, 1), 8.2 (d, 1) and 9.0 (s, 1).

Example 26 1,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- [2- (1-piperazinyl) ethylamino] azirine [21,3 ': 3,4] Pyrrolo [1,2-a] indole-10,4,7-dione carbamate This compound was otherwise prepared according to Example 1, but using dichloromethane as the solvent. Mitomycin A (50 mg) and N- (2-aminoethyl) piperazine (20 mg) gave 23 mg (36% yield) of the desired product, m.p. was 138-141 ° C (decomposed), and the analytical data are as follows: NMR (CDCl 3, TS): δ values (ppm): 6-methoxy peak 4.02 is absent and the spectrum has new peaks 1.6-2.1 (broad s , 1), 2.2 to 2.6 (broad s, 8), 2.6 to 2.8 (broad s, 4), and 6.9 (t, 1).

Example 27 1,1a, 2,8,8a, 8b-Hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- [2- (2-pyridyl) ethylamino] azirine [3,3 ': 3, 4] pyrrolo [1,2-a] indole-4,7-25 dione carbamate This compound was prepared as described in Example 1. Mitomycin A (70 mg) and an excess of 250 mg of 2- (2-aminoethyl) pyridine gave 51 mg (56% yield) of the desired product, m.p. was 64-77 ° C (decomposed), and the analytical result was as follows: NMR (CDCl 3, TS): δ values (ppm): 6-methoxy peak 4.02 missing and new peaks in the spectrum 2.8 (m, 4) 7 .0-7.8 (m, 3) and 8.5 (d, 1).

35

II

2i 80698

The above examples illustrate the following structural changes: 1. Compounds in which Z is a hydroxy-substituted 1-pyrrolinyl radical represented by Example 1.

2. Compounds in which Z is a lower alkyl-substituted piperidyl radical represented by Example 2.

3. Compounds in which Z is substituted by acetamino, acetyl, carbamido, cyano, carboxy (lower alkyl), di- (lower alkoxy), nitro, sulfamyl or (lower alkyl) aniline radical, represent Examples 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 and 15, respectively.

4. Compounds in which Z is a radical of formula

R

I.

-N-R1 and wherein R1 is a nitrogen-containing heterocyclic radical which is amino-substituted triazolyl, lower alkyl-substituted isothiazolyl, benzothiazolyl or a nitro- or halogen-substituted derivative of benzothiazolyl are represented by Examples 16, 17, 18, 19, respectively.

5. Compounds wherein Z is a radical of formula

R

I.

-N-R1 wherein R1 is a substituted lower alkyl radical which is hydroxy (lower alkylamino) (lower alkyl), hydroxy- (lower alkoxy) (lower alkyl), imidazolyl (lower alkyl), (nitro-substituted pyridylamino) (lower alkyl), piperazinyl (lower alkyl) and pyridylethyl, represent Examples 21, 22, 23, 24, 25, 26 and 30 27, respectively.

Although none of the above examples illustrate compounds in which Y is other than hydrogen, compounds in which Y is lower alkyl are nevertheless within the scope of the invention; reference is made herein to U.S. Patent Nos. 4,268,676, 4,460,599 and 4,617,387.

Claims (25)

A process for the preparation of therapeutically useful derivatives of mitomycin A of the formula (x 1 L A process according to claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- (3-hydroxy-1-pyrrolidinyl) -azirino [2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with 3-pyrrolidinol. A process according to claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- (3-methylpiperidyl) -azirino [ 2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with 3-methyl-piperidine. A process according to claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- [4- (acetylamino Janilino] azirino [2 ', 3': 3,4] -pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with 4- (acetylamino) aniline. Process according to claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- [3- (acetylamino) anilino] - azirino- [2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with 3- (acetylamino) )aniline. A process according to claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- (4-acetylanilino) -azirino [2 ', 3': 3, 4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with 4-acetyl-aniline. 7. A process according to claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- [4- (ureido) analino ] -azlrino [2 ', 3': 3, 4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with 4- (1-ureido) aniline . 8. A process according to claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- [4- (cyanoanilino] -azirino) [2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with 4-aminobenzonitrile. A process according to claim 1 for the preparation of 1,1a, 2,8,8a, 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, characterized in that mitomycin A is reacted with 3-30 aminobenzonitrile. A process according to claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- [4- (N-glycyl) anilino] - azirino- [2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with 4- (N-glycyl) )aniline. Il 31 80698 10. I] I / AND I L_D> Y where Y is hydrogen or lower alkyl; and X is a hydroxy substituted 1-pyrrolidinyl radical or a lower alkyl substituted piperidinyl radical or one with acetamino, acetyl, carbamido, cyano, carboxy (lower alkylamino), di- (lower alkoxy), nitro or sulfamyl substituted aniline radical or a radical of the formula R -N-R1 wherein R is hydrogen or lower alkyl and R1 is a nitrogen-containing heterocyclic radical which is an amino substituted triazolyl, lower alkyl substituted isothiazolyl, benzothiazolyl or a nitro or halogen substituted benzothiazolyl derivative, or R 1 is substituted lower alkyl radical, which is hydroxy (lower alkylamino) (lower alkyl), hydroxy (lower alkoxy) (lower alkyl), imidazolyl (lower alkyl), (nitrosubstituted imidazolyl) (lower alkyl) , (nitrosubstituted pyridylamino) (lower alkyl), or piperazinyl (lower alkyl), characterized in reacting mitomycin A or N-alkyl-mitomycin A with an amine in in the form of pyrrolidinol, or lower alkylpiperidine, or with acetamino, acetyl, carbamido, cyano, carboxy (lower alkylamino), di- (lower alkoxy), nitro or sulfamyl substituted aniline, or with an amine or a (lower alkyl) ) substituted derivative thereof which is an amino-substituted triazolylamine, (lower alkyl) substituted isothiazolylamine, benzothiazolylamine or a nitro- or halogen-substituted benzothiazolylamine, hydroxy- (lower alkylamino) (lower alkylamine), hydroxy- (lower alkoxy) ), imidazolyl (lower alkylamine), (nitrosubstituted imidazolyl) (lower alkylamine), (nitrosubstituted pyridylamino) (lower alkylamine) or piperazinyl (lower alkylamine), and isolates the desired compound from the reaction mixture. A process according to claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- (3,4-dimethoxy-anilino) -azirino- [ 2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with 3,4-dimethoxyaniline. A process according to claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- (3,5-dimethoxy-anilino) -azirino- [ 2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with 3,5-dimethoxyaniline. A process according to claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- (4-nitroanilino) -azirino- [2 ', 3': 3,4] pyrrolo- [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with 4-nitroaniline. A process according to claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- (4-sulfamylanilino) -azirino- [2 ', 3': 3,4] pyrrolo- [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with 4-sulfanilamide. 25 A process according to claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- [5-amino-1,2,4- triazol-3-yl) amino] -azirino- [2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with 3,5-diamino-1,2,4-triazole. The process according to claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6 - [(3-methylisothiazol-5-yl) amino ] -azirino- [21,3 ': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with 5-amino 3-methylisothiazole. 32 80698 The process of claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- [(2-benzothiazolyl) amino] -azirino [2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with 2-aminobenzothiazole. The process according to claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6 - [(6-nitrobenzothiazol-2-yl) amino ] -azirino [2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with 2-amino 6-nitrobenzothiazole. 19. A process according to claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6 - [(4-chlorobenzothiazol-2-yl) amino] -azirino- [2 ', 33,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with 2-amino-chlorobenzothiazole. 20. A process according to claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- [2- (2-hydroxyethylaminoethylamino] -azirino) - [2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with 2- (2-aminoethylamino) ethanol. A process according to claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- [2- (2-hydroxyethoxy) ethylamino] - azirino- [2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with 2- (2-aminoethoxy) ethanol . A process according to claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- [2- (4-imidazolyl) ethylamino] - azirino- [21,31: 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with histamine dihydrochloride. Il 33 80698 A process according to claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6 - [(2-nitro-1-imdazolyl) ethylamino ] -azirino- [2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with 1- (2-aminoethyl) - 2-nitroimidazole. The process of claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- {2- [(5-nitro-2 pyridyl) amino] ethylamino} -azirino [2 ', 3': 3,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with 2- ( 2-aminoethyl) -5-nitropyridine. A process according to claim 1 for the preparation of 1,1a, 2,8,8a, 8b-hexahydro-8-hydroxymethyl-8a-methoxy-5-methyl-6- [2- (1-piperazinyl) ethylamino] -azirino- [233,4] pyrrolo [1,2-a] indole-4,7-dione carbamate, characterized in that mitomycin A is reacted with N- (2-aminoethyl) piperazine.