Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
252~
"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; No. 4,460,529; and my co-pending Canadian
Patent Application Serial No. 3B9,102 filed March 11,
1982 may provide material relating to the present
invention.
Briefly summarized, said UOS. Patents 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,
H ~ NH 2
wherein: Y is hydrogen or lower alkyl; and X
is a thiazolamino radical, a furfurylamino radical or a
radical of the formula,
-- 2
R ~1
---N - C - R
12
in which R, Rl and R2 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, tetrahydrofuxyl
and benzene sulfonamide.
Said U.S. 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, Ia,
0
0 11
~ ~ CH2OCNH2
\,/\
II I 10CH3 Ia
H3C~/ ~b
o
wharein: Y is hydro~en or lower alkyl; and Z
is a thiazolamino radical, a furfurylamino radical, a
cyclopropylamino radical, a pyridylamino radical, or a
radical of the formula,
R4 R5
I i 7
- N ~ - R
R
~'
~2~
-- 3
in which R4, R5, and R6 are the sa~e ox different and
selected from the group consistin~ of hydrogen and
lower alkyl, and R7 is selçcted from the group consist-
ing of lower alkenyl, halo-lower alken~l, lower alkynyl,
5 lower alkoxycarbonyl, halo-lower alkyl, hydroxy-lower
alkyl, pyridyl, thienyl, formamyl, tetrahydrofuryl,
benzyl, and benzene sulfonamide.
~ .S. Patent NoO 4,460,529 also discloses
compounds with a substantial degree of antitumor activity
10 in animals of the following formula IIa,
O O
~JI~ / CH20CNH2
lS ~ IIa
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 rom the group consistin~ of l-pyrrolinyl, 1-
indolinyl, N-thiazolidinyl, N-morpholinyl, l-piperazinyl,
and N-thiomorpholinyl radicals, or
a cyano, phenyl, carboxamido or lower alkoxy-
carbonyl substituted l-aziridin~l radical, or
a lower alkyl, formyl or acetylphenyl substituted
1-piperazinyl radical, or
an hydrox~ or pipe~idyl substituted l-piperidyl
radical, or
a lower alkoxy, amino or halo substituted
35 pyridylamino radical, or
`
a c~rboxamido, mercapto ox ~ethylenedioxy
substituted anilino radical, or
R
a radical of the for~ula, -N-R1
wherein R is hydro~en or lower alkyl and R'
is a nitrogen-containing heterocyclic radical selected
from the group consistin~ 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 ~roup 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, l-pyrrolidinyl
lower alkyl, N-lower alkyl pyrrolidinyl lower alkyl, N-
morpholinyl lower alkyl, and lower dialkylamino lower
alk~l.
Also pertinent to the back~round of the
present invention are the followin~ references: Cosulich,
et al., U.S. Patent No. 3,332,944; Matsui, et al., U.S.
Patent No. 3,410,867; ~akano, et al., U.SO Pate~t No.
4,231,936; Matsui, et al., U.S. Patent No. 3,429,894;
30 Remers, U.S. Patent No~ 4,268,676; ~atSui, et al., U.S.
Patent No. 3,450,705i Matsui, et al., U.S. Patent No~
3,514,452; and I~ai, et al., Gann, 71, pp. 560-562
(1980).
- 35
~5~
BRIEF SUMMARY
Accordin~ to the present invention, there are
provided no~el compounds of the formula, III,
O
Il 11
~CE~20CNH ;~
¦ OCH3
~ ~ NY III
wherein: Y is hydro~en or lower alkyl; and X
15 is
an hydroxy substituted l-pyrrolidinyl radical,
or
a lower alkyl substituted piperidyl radical,
or
an acetamino, acetyl, carbamido, cyano,
carboxy lower alkylamino, di-lower alkoxy, nitro, or
sulfamyl substitu~ed anilino radical, or
a radical of the formula, -N-R
wherein R is hydrogen or lower alkyl and Rl
is a nitrogen containing heterocyclic radical selected
from the group consisting o~ amino substituted triazolyl,
lower alkyl substituted isothiazolyl, benzothiazolyl,
and nitro and halo substituted derivatives of benzo-0 thiazolyl, or Rl is
a substituted lower alkyl radical selected
from the group consisting of amino lowex alkyl, lower
alkylamino lower alkyl, hydroxy lower alkylamlno lower
alkyl, hydroxy lower alkoxy lower alkyl, imidazolyl
35 lower alkyl, nitro substituted imidazolyl lower alkyl,
~2~
-- 6 --
nitro substituted pyridylamino lower alkyl, and
piperazinyl lower alkyl.
Also provided according to the invention are
novel methods for treatment of neoplastic disease states
in animals, which methods comprise administering a
therapeutically effective amount of a compound of the
formula, IIIa,
Z 11
~ / ~ ~--CH20CNH2
l ~ CH3
H3 ~ ~ ~ IIIa
wherein: Y is hydrogen or lower alkyl; and Z
is
an hydroxy substituted l-pyrrolidinyl radical,
or
a lower alkyl substituted piperidyl radical,
or
an acetamino, acetyl, carbamido, cyano,
carboxy lower alkylamino, di-lower alkoxy, nitro,
sulfamyl, or lower alkyl substituted anilino radical,
or R
a radical of the formula, -N-Rl
wherein R is hydrogen or lower alkyl and Rl is
a n.itrogen containing heterocyclic radical selected ~rom
the group consisting of amino substituted triazolyl,
lower alkyl substituted isothiazolyl, benzothiazolyl,
and nitro and halo substituted derivatives of benzo-
thiazolyl, or Rl is
a substituted lower alkyl radical selected
from the group consisting of amino lower al]cyl, lower
~2~
7 --
alkylamino lower alkyl, hyd~oxy lower alkylamino lower
alkyl, hydroxy lower alkoxy, lower alkyl, imidazolyl
lower alkyl, nitro substi~uted imidazolyl lower alkyl,
nitro substituted pyridylamino lower alkyl, piperazinyl
lower alXyl, 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 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
having one to six carbon atoms.
It will be apparent that the compounds of
formula III are all comprehended by the specifications
of formula IIIa. Put another way, all the novel anti-
biotic mitomycin derivatives of formula III are useful
in practice of the novel antineoplastic therapeutic
~0 methods which involve administration of compounds of
formula IIIa.
Mitomycin derivatives of the invention are
prepared by the reaction of mitomycin A with the
appropriately selected amine compounds. The N-alkyl-
~5 mitomycin (e.g., N-methylmitomycin) derivatives are
similarly prepared by the reaction of a selected amine
with N-alkylmitomycin h prepared from mitomycin C, e.g.,
according to the methods generally disclosed in Cheng,
et al., J.Med.Chem., 20, No. 6, 767-770 (1977). The
preparative reactions generally yield the desired
product as a crystalline solid which is readily soluble
in alcohol.
Therapeutic methods of the invention compre-
hend the administration of effective amounts of one or
more of the compounds of formula IIIa, as an active
ingredient, . . . . . . . . . . . . . . . . . . . . .
._
together with desired pharmaceutically acceptable
diluents, adjuvants and carrie~s, to ~n animal suffering
from a neoplastic disease state. Unit dosa~e 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 0.00~ 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 a~out 0.2 to about 51.2
10 mg per kg, of body weight o~ 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.
DESCRIPTIO~ OF INVENTION
The following Examples 1 through 32, descri~ing
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
~5 out at room temperature (20QC), without added heat.
Unless otherwise indicated, all thin layer chromato-
graphic (TLC) procedures emplo~ed to check the progress
of reactions involved the use of a pre-coated silica-
gel plate and a mixture o~ methanol and chloroform (2~8
30 by volume~ as a developin~ solvent~
E~
1,la,2,8,8a,8b-Hexahydro-8-(hydroxymethyl~-8a-methoxy-
5-methyl-6- 3-hydroxy-1-pyrrolidinyl)-azirino~2',3':3,4]
pyrrolo[l~2-a~indole 4,7-dione carbamate
A solution of mitomycin A (50 mg) in 6 ml of
anhydrous methanol was treated with 3-pyrrolidinol (13
mg) under nitrogen at room temperature. W~en thin-
layer chromatography on silica gel (2:8 methanol-
chloroform as solvent) showed that staxting material no
10 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-
5 85C (decomposition) and providing the following analysis:NMR (DMSO-d6, TS): '~' values in ppm.
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).0
Example_2
1,la,2,8,8a,8b-Hexahydro-8-(hydroxymethy1)-8a-methox~t-
5-methyl-6-(3-meth=ylpiperdyl)-azirino[2',3':3,4]pyrrol~o
[1,2-a~indole-4,7-dione carbamate5
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 pxoduct having a melting point of
30 75-88C (decomposition) and p~oviding the following
analysis:
-- 10 --
NMR (CDC13, TS)~ alues 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 (m,5) and 2.15-3.32 (m,4).
s
Example 3
1,la,2,S,8a,3b~Hexahydro-8-(hydroxymethyl)-8a-methoxy-
5-methyl-6~ piperazinyl)-azirino[2',3':3,4]pyrxolo-
[1,2-a]indole-4,7-dione carbamate
This compound was prepared by the procedure
described in Example 1. From 60 ~g of mitomycin A and
30 mg of anhydrous piperazine was obtained 23 mg (34%
yield) of the desired product having a melting point
15 greater than 200C (decomposition) ~nd providin~ the
following analysis:
NMR (DMSO-d6, TS): '~' values in ppm.
Absence of the 6-~ethoxy peak at 4.02 and the
appearance of new peAks at 1.9 (broad s,l)
and 2.9 (s,8).
Example 4
1,la,2,8,8a,8b-Hexahydro-8-(h dxoxymethyl~-8a-methoxy-
5-methYl-6-[4-(acetylamino)anilino]-azirino[2',3':3,4]
25 Eyrrolo[1,2-a]indole-4,7-dlone carbamate
This compound was p~epaxed by the procedure
described in Example 1. ~xom 100 m~ of mitomycin A and
excess 4-(acetylamino)aniline ~s obtained 102 ~g (76%
30 yield~ of the desired pxoduct haYin~ a melting point of
143q-145C (decomposition) and pxo~iding the ~ollowin~
analysis:
~25~
11 --
NMR (CDC13, TS): '~' values in ppm.
Absence of th~ 6-~ethoxy peak at 4.02 and the
appearance o~ 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,la,2,8,8a,8b-Hexahydro-8-(hydroxymethyl)-8a-methoxy-
5-methyl-6-[3-(acetylamino)anilino]-aæirino[2l,3':3,4]
rrolo[l 2-a]indole-4,7-dione caxbamate
F ,~ r
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 o~ 140-143~C (decomposition)
and providing the following analysis:
N~R (Acetone-d , TS): '~' values in ppm.
Absence of the 6-methoxy peak at 4.02 and the
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
25 1,la,2,8,8a,8b-Hexahydro-8-(hYdroxYmethyl)-8~-methoX -
5-methvl-6-('4-acetYlanilino)-azirinoL2',3':3-,4]pyrrolo
Ll,2-a~indole-4,7-dione caxbamate
This compound wa~ prep~red by the procedure
30 described in Example 1, except that a small amount of
solid potassium carbonate was ~dded. From 70 mg o
mitomycin A and 510 mg of 4-acet~Ylaniline was obtained
25 mg (28~ yield) of the desired product having a
melting point o 103-104C (decomposition~ and providing
-- 3i the o}1Owing analysis:
NMR (CDC13, TS), I~ valu~s in pp~.
~bsence of the 6-methoxy peak at 4.02 and the
~ppearance of ne~ peaks at 2.1 (s,3~, 6.6(d,2),
7.3 (d,2) and 7.0-7.~ (broad s,l~.
Ex'amp1e 7
1,la,2,8,8a,8b-Hexahydro-8-(hydroxymethyl)-8a-me'thoxy-5
meth~l-6-[4-(1-ureido)anilino]-azirino[~',3':3,4]pyrrolo
[1,2-a]indole-4,7-dione carbamate
_
' This carbamido-substituted compound was prepared
by the procedure described in ~xample 1. From 50 mg of
mitomycin ~ and 227 mg o~ 4-(1-ureido)aniline was obtained
49 mg (67% yield) of the desired p~oduct having a melting
15 point of ~3-95C (decomposition) and providing the fol-
lowing analysis:
NMR (CDC13, TS): '~' values in ppm.
Absence of the 6-methoxy peak at 4.02 and the
appearance of new peaks at 5.03 (s,2), 6.9
(d,2), 7.3 (d,2), 8.0 (s,l) and 8.4 (s,l).
Example 8
1,la,2,8,8a,8b-Hexahydro-8-(h~d~oxymeth 1~-8a-methox -5-
methyl-6-(4--cyanoanilino)-azirino[2',3l:3r4]pyrrolo[1,2-a3
25 indole-4,7-dione carb'amate
This compound wa~ prepared by the procedure
described in Example l, except that a small amount of
solid potassium carbonate waS added. From 70 m~ of
30 mitom~cin A and 472 mg of 4-aminobenzonitxile was
obt~ined 23 mg (24% ~ield~ of the desi~ed product
having a melting point of 124-126QC (decomposition~
and providing the following anal~sis:
- 13 -
NMR (CDC13, TS): '~ ' values in ppm.
Absence of the 6-methoxY peak 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~.
Exam~le 9
1,la,2,8,8a,8b-Hexahydro-8-(hydroxymethyl)-8a-methoxy-5~
methyl-6-(3-cyanoanilino)-azirino[2',3':3,4]pyrroloEl,2-a]
indole-4,7-dione carbamate
_ _ _ _
This compound was prepared by the procedure
described in Example 1, except tha-t a small amount of
solid potassium carbonate was added. From 71 mg of
mitomycin A and 500 mg of 3-aminobenzonitrile was
1~ obtained 30 mg (34% yield) of the desired product
having a melting point of 97-98C (deoomposition) and
providing the following analysis:
NMR (CDC13, TS): '~' values in ppm.
Absence of the 6-methoxy peak at 4.02 and the
~0 appearance of new peaks at 7.2-7.8 (m,4).
Example 10
1,la,2,8,8a-,8b-Hexahvdro-8 (hYdroxYmethyl)-8a-methoxY-5-
~- methyl-6-[4-(N-glycv ~ ,3':3,4]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
249 mg of 4-(N-glycyl)aniline was obtained 62 mg (90~
yield) of the desired product having a melting point of
83-85C (decomposition) and providing the following
analysis:
- 14 -
NMR (DMSO-d6, TS): '~' values ln ppm.
~bsence of the 6-methoxy peak at 4.02 ~nd 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
(broad s,2).
E ample 11
1,la,2,8,8a,8b-Hexahydro-8-(h~droxymethyl)-8a-methoxy-5-
methyl-6-(3,4-''dimethoxyanilino)-azirlno[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 50 mg of mitomycin A and 229
mg of 3,4-dimethoxyaniline was obtained 61 mg (91~ yield) of
15 ~he desired product having a melting point of 114-
116C (decomposition) and providing the following
analysis:
NMR (CDC13, TS): '~' values in ppm.
Aksence of the 6-methoxy peak at 4.02 and the
appearance of new peaks at 3.8 (s,6), 6.3-~.9
(m,3) and 7.7 (s,l).
Example 12
1,la,'2,8,8a,3b-Hexahydro-8-(hydroxymethyl~)-8a-methoxy-5-
~ methyl-6-(3,5-dimethoxyanilino)-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. F~om 50 mg of mitomycin A and
30 229 mg of 3,5-dimethoxy~niline was obtained 60 mg (88~
yield) of the desired product having a meltin~ point of
98-100C ~decomposition) and providing the following
analysis:
-- NMR tcDcl3~ TS): '~' values in ppm.
Absence of the 6-methoxy peak at 4.02 and the
appearance of new peaks at 3.8 (s,6~, 5.9-6.4
(broad s,3) and 7.6 (s,l~.
Example 13
1,la,2,8,8a,8b-Hexahydro-8-(hydroxymethyl)-8a-methox~-5-
meth~l-6-(4-nitroanilino)-a~irino~2',3':3,4~p~rrolo[1,2--a]
indole-4,7-dione carbamate
Th.is 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 the5 follo~ing analysis:
NMR (Acetone-d6, TS): '~' values in ppm.
Absence of the 6-methoxy peak at 4.02 and the
appearance of new peaks at 6.9-7.3 (d,2),
7.4-i.9 (d,2) and 7.9-8.4 (broad s,l).
Example 14
1,la,2,8 t 8a,8b-Hexahydro-8-(hydroxymethyl)-8a-methoxy-5
methyl-6-(4-sulfamylan lino)-azirlno[2' ! 3' 3,4]pyrrolo
[1,2-a]indole-4,7-dione carbamate
~5
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 o
mitomycin A and 688 mg of sul~anilamide was obtained 25
mg (26% yield) of the desired product having a melting
point of 113-115C (decomposition) and providing the
following analysis:
NMR (CDC13, TS): '~' values in ppm.
Absence of the 6-methoxy peak at 4.02 and the
appearance o~ new peaks at 7.0 (d,2), 7.5
(s,l) and 7.9 (d,2).
~2~'7~5~
- 16 -
Example 15
1,la,2,8,8a,8b-Hexahydro-8-(hydroxymethyl~-8a-~ethoxy-5-
metnyl~6-(4-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 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-
115C (decomposition) and providing the followinganalysis:
NMR (CDC13, TS): '~' values in ppm.
Absence of the 6-methoxy peak at 4.02 and the
appearance of new peaks at 2.3 (s,3), 6.5-7.3
(broad s,4) and 7.6 (broad s,l).
Example 16
1,la,2,8,8a,8b-Hexahydro-8-(hydroxymethyl')-8a-methoxy-5-
methyl-6-(3-methylanilino)-azirino[2',3':3,4]pyrrolo~1,2-a]
indole-4,7-di`one 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%
~5 yield) of the desired product having a melting point of
89-91C (decomposition) and providing the following
analysis:
NMR (CDC13, TS): '~' values in ppm.
Absence of the 6-methox~ peak at 4.02 and the
appearance o~ new peaks at 2.4 (s J 3), 6.7-7.5
~m,4) and 7.8 (5,1).
Example 17
1,la,2,8,8a,'8b-Hexahydro-8-(hydroxymet~lyl~-8a-methoxv-5-
methvl-6-[(5-amino-1,2,4-triazol-3-yl~amino]-a~irino
[2',3':3',4~pyrro'lo['1,'2-a'lindole-4,7-dione carbamate
'7
- 17 -
This compound was prepared by the pro-edure
described in Example 1, exce~t that a sm~11 amount of
solid potassium carbonate was added. From 50 mg of
mitomycin A and 30 mg of 3,5-diamino-1,2,4-tria~ole was
obtained 13 mg (5.5O- yield) of the desired product
having a melting point of 117-120C (decomposition)
and providing the following analysis:
NMR (DMSO-d I TS): '~' values in ppm.
Absence of the 6-methoxy peak at 4.02 and the
appearance of a new peak at 5.37 (s,3).
Example 18
1,la,2,8,8a,8b-Hexahydro-8-(hydroxymethyl)-8a-methox~
methx1-6-[(3-methylisothiazol-5-yl)amino]-azirino[2',3':3,4]
pyrrolo[l,2-a]indole-4,7-dione caxbamate
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-
a~ino-3-methylisothiazole hydrochloride was obtained
~.5 mg (8.5~ yield) of the desired product having a
melting point of 87-90C (decomposition) and providing
the following analysis:
N~R (CDC13, TS): '~' values in ppm.
Absence of the 6-methoxy peak at 4.02 and the
appearance of new peaks at 2.3 (s,3~, 6.1
~s,l) and 6.4 (s,l).
Example 19
1,la,2,8,8a,8b-Hexah~dro-8'-(hydroxymethyl)-8a-met~oxy-5-
meth~l-6-[l2-benzothiazolyl~amino]-azlrino[2',3':3,4]_yrrolo
[1,2-a]indole-4,7-dione carbamate
~ .
This compound was prepared by the procedure
described in Example 1, except that a small amowlt of
solid potassium carbonate was added. From 50 mg of
~'~52~
- 18 -
mitomycin A and 5 mg of 2-aminobenz~thiazole was
obtained 12 mg (18% yield~ of the desired p~oduct
having a meltirIg point of 82-85PC (decomposition~ and
providing the following analysis:
NMR (CDC13, TS) '~' 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
1,la,2,8,8a,8b Hexahxdro-8-(hydroxymethyl)-8a-methoxy
5-methyl-6-[(6-nitrobenzothiazol-~-~l)amlno]-azirino
[2',~3i.3,4]pyrrolo[1 ! 2-a]indole-4,7-dione carbamate
This compound was prepared by the procedure
15 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-nitrobenzothiazole
was obtained 20 mg (27~ yield) of the desired product
having a melting point of 86-89C (decomposition) and0 providing the following analysis:
NMR (DMSO-d , TS): '~' values in ppm.
Absence of the 6-methoxy peak at 4.02 and the
appearance of new peaks at 6.9-8.3 (m,4).
Example 21
1,la,2,8,8a,8b-Hexahydro-8-(hydroxymethyl)-8a-methoxy-5-
methyl-6-[(4-chlorobenzothiazol-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 150 mg of
mitomycin A and 27 mg of 2-amino-chlorobenzothiazole
was obtained 30 mg (14% yield) of the desired product
35 having a melting point of 89-91C (decomposition) and
providing the following analysis:
-- 19 --
N~R (CDC13, TS)o '~' values in pp~
Absence o~ the 6-methoxy peak at 4.02 ~nd the
appearance of new peaks at 7.1-8.0 (broad
s , ~ ) .
Example 22
1,la,2/8,8a,8b-Hexahydro-8-(hydroxymethyl)-8a-methoxy~5-
methyl-6-~(2-aminoethyl)amino~-azirino[2',3':3,4]pyrrolo
~1,2-a~indole-4,7-dione carbamate
a '- -~-~ ~---~~'- ~~'
This compound was prepared by the procedure
described in Example 1, except that the solvent was
dichlorome-~hane. From 50 mg of mitomycin A and 10 mg
of 1,2-diaminoethane was obtained 35 mg (65% yield) of
1~ the desired product having a melting point of 202-
205C (decomposition) and providing the following
analysis:
NMR (CDC13, TS): '~' values in ppm.
A~sence of the 6-methoxy peak at 4.02 and the
appearance of new peaks at 1.5 (broad s,2)
and 3.5 (broad s,4).
25 1llal2l8`l8al8b-Hexahydro-8-(hydroxymethyl)-8a-methoxy-5
methyl-6-[methyl(2-methylaminoethyl)amino]-azirino
[2',3':3,4]pyrrolol1,2-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 ~g of sym-dimethylethylenediamine
was obtained 28 mg (50~ yieldl of the desired product
having a melting point of 99-101C (decomposition~ and
providing the following analysis:
- 20 -
NMR (CDC13, TS)~ values in pp~.
Absence of the 6-methox~ peak at 4.02 and the
appearance of new peaks at 1.3 (s,].), 2.5
(s,6), and 2.7 (s,4).
Exam~le 24
1,la,2,8,8a,8b-Hexah dro-8-(hydroxymethY1)-8a-methoxy-5-
methyl-6-[2-(2-hydroxyethylamino)ethy-lamïno]-azirino
[2'~,3'.3,4]pyrrolo[1,2-a]indole-4,7-dione carbamate
1~
This compound was prepared by the procedure
described in Example 1, except that the solvent was
dichloromethane. From 50 mg of mitomycin ~ and 18 mg
of 2-(2-aminoethylamino)ethanol was obtained 35 mg (58~
15 yield) of the desired product having a melting point of
115~-118C (decomposition) and providing the ollowing
analysis:
NMR (CDCl , TS): '~' values in ppm.
Absence of the 6-methoxy peak at 4.02 and the
~0 appearance of new peaks at 2.7 (broad s,7)
and 3.7 (t,3~.
Exam~le 25
~5 1,la,2,8,8a,8b-Hexahy~dro-8-(hydroxymethyl)-8a-meth~y~
methyl-6-[2-_(2-hydroxyethoxy)ethylaminol-azi_ino[2',3l:3,4]
pyrrolo[l,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 mitom~cin A and 20 mg
of 2-t2-aminoethoxy)ethanol was obtained 30 m~ (42%
yield) of the desired product having a melting point of
99-102C (decomposition) and providing the ~ollowing
analysis:
- 21 -
NMR (CDC13, TS): '~' values in pp~.
Absence of the 6-methoxy peak at 4.02 and the
appearance of new peaks at 3.3-3.9 (broad
s,9) and 6.4-6.8 (broad s,l).
Example 26
1r la,2,8,8a,8b-Hexahy~ro-8-(hydroxymethyl)-8a-metho~y-5-
methyl-6-~2-(4-imidazolyl)ethylamino]-azirino[2',3':3l4]
pyrrolo[l,2-a]ind'ole-4!7~dione carb~amate
This compound was prepared by the procedure
described in Example 1, except that 128 mg of sodium
methox~de was added. From 70 mg of mitomycin A and 368
mg of histamine dihydrochloride was obtained 61 mg (71~
15 yield) of the desired product having a melting point of
72-73C (decomposition) and providing the following
analysis:
NMR (DMSO-d6, TS~ ' values in ppm.
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).
E_ample 27
25 1,la,2,8,8a,8b-Hexahydro-8-(hydroxymethyl)-8a-methoxy-5-
methYl-6-[(2-nitro-1-imidazolyl)ethylamino]-aziri'no' ' '~
~2',3':3,4jpyrrolo[1,2-a]indole-4,7-dlone carbamate
This compound was prepared by the procedure
described in Example 1. From 72 mg o~ mitomycin A and
30 excess 1-(2-aminoethyl)-2-nitroimidazole was obta~ned
60 mg (70% yield) of the desired product havin~ a
melting point of 83-85C (decomposition) and provlding
the following analysis:
_ NMR (CDC13, TS): '~' values in ppm.
. 35 Absence-of the 6-methoxy peak at 4.02 and the
~5~
- 22 -
appearance of new peaks at 3.4 (t,2), 4.6
(t,2), 7.3 (broad s,2) and 7.6 (s,l~.
Example 28
1,la,2,8,8a,8b-Hexahydro-8-(hydroxymethyl)-8a-methoxy-5-
methy__6-[2-(4-hydrox~phenyl)ethylamino]-azirino[2',3':3,4]
pyrrolo[l,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 of tyramine was obtained 138 mg t81% yield) of
the desired product having a melting point of 120-
125C (decomposition) and providing the following
analysis:
NMR (CDC13, TS): '~' values in ppm.
~bsence 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 (d,2), 7.0 (d,2) and 8.0 (s,l).
Example 29
1,la,2,8,8a,8b-Hexah~dro-8-(hydroxymethyl)-8a-methoxy-5-
methyl-6-[2-(3,4-dihydroxyphenyl)ethy~_ino]-a~irino
[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 hydrobromide was obtained
60 mg (40% yield) of the dssired product decomposing
30 without melting above 125C and providing the following
analysis:
NMR (CDC13, TS): '~' values in ppm 7
Absence o the 6-methoxy peak ~t 4.02 and the
appearance of new peaks at 2.6 (t,2), 2.8
(t,2), 6.4-6.8 (m,3) and 8.3 (broad s,2~.
~:25~
Example 30
1,la,'2,8,8a,8b-Hexahydro-8-(hydroxymethyl)-8a'-methoxy-5-
methyl-5-~2-[(5-nitro-2-pyridyl~amino]ethylamino'~-azirlno
[2',3':3,4~pyrrolo[1,2-a]indole-4,7~dione carbamate
This compound was prepared by the procedure
described in Example 1, e~cept that the solven~ was
dichloromethane. From 50 mg of mitomycin ~ 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 76-79C (decomposition) and providing the
following analysis:
NMR (CDC13, TS): '~' values in ppm.
Absence of the 6-methoxy peak at 4.02 and the
appearance of new peaks at 3.3-4.0 (m,43,
6.2-6.7 (broad s,2), 8.1 (d,l), 8.2 (d,l) and
g.0 (s,l).
Example 31
1,la,2,8,8a,8b-HexahYdro-8-(hydroxYmethyl')-8a-me`thoxy-5-
methyl-6-[2-(1-pipérazinyl)ethylamino]-azir-ino[2',3'-3,4]
E~rrolo[1,2-a]indole-4,7-dione carbamate
This compound was prepared by the procedure
~5 described in Example 1, except that the solvent was
dichloromethane. From 50 mg of mitomycin ~ ~nd 20 mg of
N-(2-aminoethyl) piperazine was obtained 23 mg (36%
yield) of the desired product having a melting point o~
138-141C (decomposition) and providing the following0 analysis:
N~R (CDC13, TS): '~' values in ppm.
Absence o~ the 6-methox~ 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
s,4) and 6.9 (t,l).
s~
24 -
Example 32
1,la,2,8,8a,8b-Hexahydro-8-(hydroxymethyl)-8a-methoxy 5
methyl-6-L2 (2-pyrid~l)ethyla~ino]-azixinoE2',3':3,4
pyrrolo~l,2-a]indole-4,7-dione carbamate
-
This compound was prepar2d 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~ o~ the desired product havin~ a
10 melting point of 64-77C (decomposition) and providing
the following analysis:
NMR (CDCl , TS): '~' 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
1~ (m,3) and 8.5 (d,l).
With specific reference to the compounds
comprehended by formula IIIa, the above examples illustrate
the following structural variations.
~0 1. Compounds wherein Z is a hydroxy substituted
l-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
25 radical or an acetamino, acetyl, carbamido, cyano,
carboxy lower alkylamino, di-lo~er 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 ~herein Z is a radical of the
formula
R
_N_~l and wherein Rl is a nitro~en containin~ heterocylic
radical selected from the group consistin~ o~ amino
35 substituted triazolyl, lower alkyl substituted isothiazolyl,
benzothiazolyl and nitro and halo substituted derivatives
~25~
- 25 -
of benzothiazolyl are represented, respectively, ~y
Examples 17, 18, 19, 20 and 21.
5. Compounds wherein Z is a radical of the
formula
R
-N-Rl and wherein Rl 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
alk~l, imidazolyl lower alkyl, nitro substituted
imidazolyl lower alkyl, mono- and di-hydroxy phenyl
lower alkyl, nitro substituted pyridylamino lower alXyl,
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
nonetheless within the comprehension of the invention,
reference being made to analogously substituted
compounds of my aforesaid U~S. Patent No. 4,268,676 and
U.S. Patent Nos. 4,460,599 and 4,617,389.
Compounds according to the present invention
are believed to possess anti-bacterial activity against
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 IIIa in the
antineoplastic therapeutic methods of the invention is
demonstrated by the results of in vivo screening
procedures wherein the compounds are administered in
varying dosage amounts to mice in which a P388 leukemic
condition is induced. The procedures were carried out
according to "~ymphocytic Leukemia P388 - Protocol
g.~;
- 26 -
1.200", published in Cancer Chemotherapy Repoxts, Part
3, Vol. 3, No. 2, page 9 (September, 1972). Briefly
put, the screening procedures involved administration
of the test comp~und to CDFl female mice previously
5 infected with 106 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 _ vivo P388 procedure noted above, a % T/C
value of 125 or greater indicates significant anti-
neoplastic 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 ~ST ~alues 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.
- 35
~2~d~9
- 27 -
TABLE 1
ExampleOptimal Dose~lST MED
No.mg/kg as % T/C
1 25.6 153 0.8
2 25.6 238 ~0.2
3 12.8 200 0.2
4 25.6 ~333 ~0.2
~5.6 231 0.2
6 6.4 167 0.4
7 25.6 194 1.6
8 3.2 150 0.8
9 12.8 172 ~0.2
10 25.6 322 0.8
11 12.8 > 333 0.2
12 6.4 161 0.4
13 3.2 172 ~002
14 25.6 225 0.2
15 12.8 167 0.4
~0 16 12.8 181 0.4
17 12.8 181 1.6
18 25.6 169 0.8
19 25.6 150 12.8
20 25.6 128 25.6
21 25.6 144 1.6
22 3.2 178 0.4
23 25.6 133 12.8
24 12.8 133 12.
25 25.6 181 0.4
26 25.6 ~ 63 1.6
27 25.6 150 3. ~
28 25.6 218 1.6
29 12.8 139 12.8
30 12.8 144 6.4
_ 35 31 25.6 138 12.8
32 25.6 > 375 0.2
~2~
-- 28 -
Clearly amon~ the most preferred compounds
employed as antineoplastic agents accoxding to the
invention are those exhibiting moxe 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 elass of such compounds is seen to include the
eompounds of Examples 4, 10, 11 and 32.
As may be noted from Table I, initial single
10 dosa~es of as little as 0.2 mg/kg showed substantial
long term antineoplastic activity. Accordingly, the
methods of the invention may involve ~herapeutic administra-
tion 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 phar-
maceutical 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
~0 suffering from neoplastic disease. It is preferred
that the compounds be administered parenter~lly.
Ph~rmaceutical compositions suitable for use in practice
o~ methods o~ the invention may eomprise simple water
solutions of one or more o~ the compounds of formula
~5 ~IIa, but may also include ~ell known pharmaceutieally
acceptable diluents, adjuvants and/or carriers sueh as
saline suitable for medicinal use.
Further aspects and advanta~es o~ the present
invention are expected to oecur to those skilled in the
30 ar~ upon consideration of the ~oregoin~ descxiption and
consequently only sueh limitations ~s appeax in the
appended claims should be pl~ced thereon.
_ 35
