Note: Descriptions are shown in the official language in which they were submitted.
The prësent invention relates to novel aminoalkyl sub- .
S stituted nitrosourea derivatives as well as their acid
addition salts with pharmaceutically acceptable acids
showing pronounced antineoplastic effectiveness in
~nimal experiments.
.; .
~ he novel compounds of the present invention may be re-
presented by the following formula:
~70
X~ CH2-NH-CO~N-CH2-CH2Cl
Y ~lkylene ~ N
'
. ' '-. ^.,~ . .
~.
, . 2
Z183
wherein X and Y are the same or different, and are
selected from the group consisting of a phenyl group ~nd
.a cyclohexyl group, sald phenyl and cyclohexyl groups
being optionally substituted with one or two substituents
selected from a halogen atom, a lower alkyl group having
from one to four carbon atoms inclusive, a trifluore methyl
group, a cyano group, a phenyl group, a cyclohexyl group
and a lower alkyloxy group having from one to four carbon
atoms inclusive.
~Alkylene" is an alkylene group, branched or unbranched,
having from one to four carbon atoms inclusive, and
Rl. and R2 are the same or different, and are each
selected from the group consisting of lower alkyl groups
having from one to four carbon atoms inclusive, and benzyl
groups, or they form together with the nitrogen atom a
saturated five- or six-membered heterocyclic ring,
. such as a pyrrolidine,-piperidine, morfoline, thio-
: morfoline, or N-lower-alkyl-piperazine ring, said hetero-
cyclic ring being optionally substituted with lower alkyl
- 20 groups having from one to four carbon atoms inclusive,
as well as pharmaceutically acceptable acid addition salts
thereof.
Background of the Invention
_ . . .. _ ..
.
.In recent years the compounds 1-(2-Chloroethyl)-3-cyclo-
hexyl-l-nitrosourea (lomustine, CCNU) and 1,3-bis(2-
chloroethyl)-l-nitrosourea (BCNU) have been found
ll~Z183
effective as cytostatic agents in the treatment of various
tumors, either alone or in combination with other cytostatic
drugs.
Unfortunately, they also have serious untoward side effects
and are rather toxic compounds, and the therapeutic index,
the ratio between the toxic dose and the therapeutic dose,
is not very favorable.
Summary of the Invention
According to the present invention it has now surprisingly
been found that the compounds of Formula I, as well as
their acid addition salts in animal experiments show
cytostatic effects comparable to BCNU and, at the same time,
less toxicity.
Based upon the animal experiments the preferred compounds
of this invention are those of Formula I, in which X and Y
are unsubstituted phenyl groups or phenyl groups having
fluorine atoms at the para-position, "Alkylene" is a
trimethylene group, optionally substituted with a methyl
group in the 2-position, and Rl and R2 are methyl groups.
The present invention also includes pharmaceutically acceptable
acid addition salts of the compounds of Formula I. Such salts
are easily prepared by methods known to the art.
The base is reacted with either the calculated amount of
~l~Z1~33 4
organic or inorganic acid in an aqueous miscible solvent,
such as acetone or methanol, with isolation of the salt by
concentration and cooling or an excess of the acid in aqueous im-
miscible solvent,such as ethyl ether or methylene chloride,with the
desired salt separating directly. Exemplary of such organic
salts are those with maleic, fumaric, benzoic, ascorbic,
embonic (pamoic) , succinic, oxalic, bis-methylene-salicylic,
methanesulfonic, ethanedisulfonic, acetic, propionic, tartaric,
salicylic, citric, gluconic, lactic, malic, mandelic, cinnamic,
citraconic, aspartic, stearic, palmitic, itaconic, glycolic,
p-aminobenzoic, 1utamic, benzene sulfonic and theophylline
acetic acids as well as the 8-halotheophyllines, for example
8-bromotheophylline. Exemplary of inorganic salts of the
compounds of Formula I are those with hydrochloric, hydro-
bromic, sulfuric, sulfamic, phosphoric and nitric acids.Of course, these salts may also be prepared by the classical
method of double decomposition of appropriate salts which is
wellknown to the art.
The compounds of Formular I and the non-toxic acid addition
salts thereof may be administered both orally and parenterally,
for example in the form of tablets, capsules, powders, syrups
or solutions for injection.
The present invention moreover comprises a method for the
preparation of the novel nitrosourea derivatives of Formula I
whereby a compound of the following formula:
X~ ~CH2-NH-CO-NH-CH2-CH2Cl
/C \ ",Rl II
Y Alkylene N ~ R2
wherein X, Y, Rl, R2 and "Alkylene" are as defined above,
is nitrosated in acid medium to yield the compound of Formula I
which is isolated either as the free base or in the form of a
pharmaceutically acceptable acid addition salt thereof.
.
'
2i~3 ; ~ 5
The nitrosation may acc~rding to the inventlon preferably
be carried out in well-known manner by using sodiumnitrite
~n acid medium, such as formic acid solution, hydrochloric
acid solution, glacial acetic solution, or the like.
Sometimes it has been found advantageous to use the NO2 -anion
formed by reduction of the NO3 -anion in situ by means of copper
in for example glacial acetic acid solution.
- The starting materials of Formula II are also novel compounds
and form part of this invention. They may according to the
invention conveniently be prepared according to the following
scheme:
X\ /CN Rl
f + Cl alkylene - N < NaNH2
Y H
~C / Rl
Y Alkylene N < L~AlH4, AlC13
X~ ~CH2NH2 Rl ..
Y Alkylene N ~ III ClCH?CH? CNO
X \ ~CH2 NHCONHCH~CH2Cl
15~ C\ Rl II
Y Alkylene N < 2
.
In all these reactions are X 9 Y ~ Rl, R2 and "~lkylene" as
defined above.
., .
-5-
11421b3
The preparation of some of the compounds of Formula III is
described in German Patent No. 2438965, and others may be
prepared in analogous manner.
The following examples are given to illustrate the method
of the present invention but, they are to be understood
as exemplary only and are not to be construed as limiting.
. ) 7
1~21.E~3
Example 1. N-t2-Chloroethyl)-N'-(2,2-bls(4-fluorophenyl)-
5-(dimethylamino)pentyl)-N-nitrosourea and its
oxalate (Lu 16-035).
i The starting material, N-(2-Chloroethyl)-N'-(2,2-b~s(4-fluoro-
phenyl)-5-(dimethylamino)pentyl)urea, was prepared in the
following way:
112 Grams 0.5 mole of bis-(4-fluorophenyl)acetonitrile were
dissolved in dry toluene and 50 grams of a 50% suspension of
soaamide in toluene were added at once while stirring,
whereupon 70 grams of 3-chloro-N,N-dimethylprop~lamine were
added, and the reaction mixture was warmed to 60-degrees Centi-
grade, where a reaction started causing a rise in temPerature
to 90 degrees Centigrade, at which temperature the reaction
mixture was kept for 30 minutes. The reaction mixture was
poured into 1 liter of crushed ice. The aqueous phase was
separated off and extracted twice with 250 milliliters of
2N hydrochloric acid. The combined aqueous phases were made
- alkaline with sodium hydroxide solution (28%), the base which
separated out was extracted with methylene chloride, the
methylere chloride solution dried over anhydrous magnesium
sulfate, evaporated and distilled. The fraction obtained
at 165-175 degrees Centigrade/0.2mmHg consisted of almost pure
2,2-bis-(4-fluorophenyl)-5-(dimethylamino)pentanenitrile.
Yield: 129 grams.
25 Grams of lithium a~minium hydride were suspended in 500
mllliliters of diethyl ~her and 82 ~rams of anhydrous aluminium
chloride in 250 milliliters of dieth~ ether added, whereupon
a solution of 125 grams of the nitrile in 250 milliliters of
~ di~thyl ether was added dropwise while stirring during 30 minutes
under reflux . The reaction mixture was boiled under reflux
~or further 3 hours and hydrolysed by adding 500 milliliters
of sodium ~ydroxide solution (28%) dropwise over 30 minutes.
The etherphase was decanted and the residue washed twice with
50Q milliliters of diethyl ether. The combined etherphases
were extracted twice with 250 milliliters of 2N hydrochloric
acid and the combined aqueous phases made alkaline to pH 10
with sodium hydroxide solution (28%). The base which separated out
, ~ ~ " ~ .
-7-
., :
~ ` 1142183
was extracted with lOOOmilliliters of diethyl ether, the ether
solution dried over anhydrous magnesium sulfate and evaporated
on a heating bath at 50 degrees Centigrade. The residue was
dissolved in 500 ml acetone/ethanol, 99% (1:1), and oxalic
acid was added until pH 3. By standing at zero degrees Centi-
grade 50 grams of the oxalate of N5, N5-dimethyl-2,2-bis(4-
fluorophenyl)-1,5-pentanediamine crystallized out. M.P.: 85-
87 degrees Centigrade.
30 Grams of this oxalate were dissolved in 100 milliliters of
water, the solution made alkaline to pH 10 with sodium hydroxide
(28%), and the base which separated out extracted twice with
100 milliliters of methylene chloride. The combined methylene
chloride phases were dried over anhydrous magnesium sulfate,
filtered and to the resulting solution were added dropwise
at zero degrees Centigrade and while stirring 10 grams of
2-chloroethyl isocyanate, whereupon the stirring was continued
for further 30 minutes. The precipitate whicn crystallized out
consisted of 22 grams of N-(2-chloroethyl)-N'-(2,2-bis(4-fluoro-
phenyl)-5-(dimethylamino)pentyl)urea melting at 205-210 degrees
Centigrade after washing with diethyl ether.
13.6 Grams of this substance were dissolved in 100 milliliters
of 2N hydrochloric acid, the solution cooled to zero degrees
Centigrade, and a solution of 4 grams of sodium nitrite in
15 milliliters of water added dropwise at 0-5 degrees Centi-
grade, whereupon the solution was stirred for further 30 minutesat zero degrees Centigrade. 100 Milliliters of methylene chloride
were added, solid sodium carbonate added to pH 8, and the
reaction mixture stirred for further 30 minutes at zero degrees
Centigrade. The methylene chloride phase was separated, dried
over anhydrous magnesium sulfate and evaporated at 20 degrees
Centigrade. The resulting oil was dissolved in 100 milliliters
of acetone and oxalic acid added to pH 3. The resulting solution
was left standing at zero degrees Centigrade and the resulting
crystals sucked off. The crystals were dissolved in boiling
methanol, and after cooling were added 100 milliliters of
dry diethyl ether.
By standing and cooling at zero degrees Centigrade 6.9 grams of
the oxalate of N-(2-chloroethyl)-N'-(2,2-bis(4-fluorophenyl)-5-
.,
(dimethylamino)pentyl)-N-nitrosourea were obtained as white
crystals melting at 144-145 degrees Centigrade.
114Z183
Example 2. N-(2-chloroethyl)-N'-(2,2-diphenyl-5-(dLmethyl-
amino)pentyl)-N-nitrosourea a~ its oxalate (Lu15-132).
The starting material, N-(2-chloroethyl)-N'-~2,2-diphenyl-5-
(dimethylamino)penty})urea, was prepared in the following way:
170 Grams of N5~Ns-dimethyl-2~2-diphenyl-I~s-pentanediamine
were dissolved in 500 milliliters of methylene chloride
and the solution cooled to zero degrees Centigrade in a
cooling bath. 70 Grams of 2-chloroeth~ isocyanate were then
added dropwise at this temperature and the reaction mixture
stirred at 30 minutes. The mixture was then evaporated at
50 degrees Centigrade, the residue dissolved in 500 milliliters
ethyl æetate/diisopropyl ether(l:l). By standing and cooling
at zero degrees Centigrade 150 grams of N-(2-chloroethyl)-N'-
((2,2-diphenyl)-5-(dimethylamino)pentyl)urea were obtained as
white crystals melting at 110-112 degrees Centigrade.
23.2 Grams of the thus obtained urea~derivative were nitro-
sated with 6~9 grams of sodiu~ ~trite in hydrochloric acid
solution as described in Example 1, and 20 grams of the
oxalate of N-(2-chloroethyl)-N'-(2,2-diphenyl-5-(dimethyl-
amino)pentyl)-N-nitrosourea were obtained as white crystals
melting at 138-140 degrees Centigrade.
.
Example 3. N-(2-chloroethyl)-N'-(2,2-diphenyl-4-methyl-5-
(dimethylamino)pentyl)-N-nitrosourea (Lu 16-022).
The starting material, N-(2-chloroethyl)-N'-((2,2-diphenyl)-
4-methyl-5-(dimethylamino)-pentyl)urea, was prepared in the
following way:
; 118 Grams of N5,N5-dimethyl-2,2-diphenyl-4-methyl-1,5-
pentanediamine were dissolved in 300 milliliters of methylene
chloride, the solution cooled to 0-5 degrees Centigrade and
' ~ 3050 grams of 2-chloroethyl ~ocyanate added dropwise while
stirring. After standing for some time at zero degrees Centi-
grade 115 grams of N-(2-chloroethyl)-N'-((2,2-diphenyl)-4-
methyl-S-~dimethylamino)pentyl)urea crystallized which, after
washing with diethyl ether, melted at 138-140 degrees Centigrade.
i~21~3
, ) ,) 10
.
25 Grams of the thus obtained urea-derivative ~ere nitro-
sated with 7 grams of sodium nitrite in hydrochloric acid
solution as described in Example 1, whereby 25 grams of N-
(2-chloroethyl)-N'-(2,2-diphenyl-4-methyl-5-(dimethylamino)
pentyl)-N-nitrosourea were obtained after crystallization
from diisopropyl ~her as yellow crystals which melted at
105-107 degrees Centigrade.
Exam~le 4. N-(2-chloroethyl)-N'-(2,2-bis(4-fluorophenyl)-
4-methyl-5-(dimethyiamino)pentyl)-N-nitrosourea.
When Example 1 was carried out using 3-chloro-N,N,2-trimethyl-
propylamine inste~d of 3-chloro-N,N-dimethylpropylamine there
were obtained as intermediates:
2,2-bis(4-fluorophenyl)-4-methyl-5-(dimethylamino)pentanenitrile
boiling at 165-175 degrees Centigrade/o.2 mmHa.
N5,N5-dimethyl-2,2-bis(4-fluorophenyl)-4-methyl-1,5-pentane-
diamine, dioxalate. M.P.: 206-208 degrees Centigrade.
N-(2-chloroethyl)-N'-(2,2-bis(4-fluorophenyl)-4-methyl-5-(di-
methylamino)pentyl)urea. M.P.: 132-134 degrees Centigrade.
By nitrosating the last mentioned urea-derivative N-(2-chloro-
ethyl)-N'-(2,2-bis(4-fluorophenyl)-4-methyl-5-(dimethylamino)
pentyl)-N-nitrosourea was obtained as a crystalline substance
melting at 105-107 degrees Centigrade.
In the same manner were the following nitrosourea derivatives
of Formula I prepared:
' ' '' ' ' . ' . `
. ~ ' '
~; .
1 0- '
. .
;'~.'c, , ~.
~ ~",,
~1~21~93
lo a
N-(2-chloroethyl)-N'-(2,2-dicyclohexyl-5-(dimethylamino)pentyl)-
N-nitrosourea maleate.
,.
S N-(2-chloroethyl)-N'-(2,2-bis(~-chlorophenyl)-5-(dimethylamino)
pentyl)-N-nitrosourea.
N-(2-chloroethyl)-N'-(2,2-bis(l-fluorophenyl)-5-(dimethylamino)
pentyl)-N-nitrosourea hydrochloride.
N-(2-chloroethyl)-N'-(2,2-bis(4-fluorophenyl)-5-(di-n-propyl-
amino)pentyl)-N-nitrosourea.
N-(2-chloroethyl)-N'-(2,2-bis(4-fluorophenyl)-5-(N-piperidine)
pentyl)-N-nitrosourea.
N-(2-chloroethyl)-N'-(2,2-bis(4-fluorophenyl)-5-(N-pyrrolidino)
pentyl)-N-nitrosourea.
N-(2-chloroethyl)-N'-(2,2-bis(4-methylphenyl)-4-(dimethylamino)
butyl)-N-nitrosourea pamoate.
N-(2-chloroethyl)-N'-(2,2-bis(4-trifluoromethylphenyl)-6-(N-
morfolire)hexyl)-N-nitrosourea.
N-(2-chloroethyl)-N'-(2,2-bis(4-methoxyphenyl)-5-(dimethylamino)
pentyl)-N-nitrosourea.
N-(2-chloroethyl)-N'-(2,2-bis(4-cyclohexylphenyl)-5-(dimethyl-
amino)pentyl1-N-nitrosourea.
ll~Z1~3 11
The following compounds have been tested in vivo in mice
according to well recognized and reliable test methods
aginst tumors:
N-(2-chloroethyl)-N'-(2,2-bis(4-fluorophenyl)-5-(dimethyl-
amino)pentyl)-N-nitrosourea, called Lu 16-035 for short, and
N-(2-chloroethyl)-N'-(2,2-diphenyl-4-methyl-5-(dimethylamino)
pentyl)-N-nitrosourea, called Lu 16-022 for short.
Lu 16-035 and Lu 16-022 were tested against the following
tumors:
10 P 388 lymphocytic leukemia i.p. implanted
L 1210 lymphoid leukemia i.p. and intracerebrally
Colon adenocarcinoma 26 i.p.
Lewis lung carcinoma i.v.
B 16 melanocarcinoma i.p.
Ependymoblastoma intracerebrally implanted
In all these tumor systems,Lu 16-035 and Lu 16-022 have shown
a significant to good effect. In some of the tumor systems
the compounds in question have been compared with BCNU (1,3-
bis(2-chloroethyl)-1-nitrosourea).
.
1. L 1210 i.p. The drugs are administered intraperitonally
(i.p.) in a single dose on day one after the
implantation of the tumor in the mice.
Lu 16-035 Survivors/total
mg/kg on day 60
200 0/7 toxic dose
100 4/7 highest non-toxic dose
3/7
; 25 4/7
12.5 0/7
8CNU mq/kq
32 3/10 highest non-toxic dose
24 0/10
-" 11421~33
12
2. Colon adenocarcinoma 26 i.p. The drugs are administered
i.p. on day 1.5 and 9 after
the tumor implantation in the mice.
(T/C % = mean survival time of test animals in percent of control)
Lu 16-035 T/C %Survivors/total
mg/kg on day 60
100 41 2/10 toxic dose
255 9/10 highest non-toxic dose
255 9/10
12.5 221 4/10
Lu 16-022 mg/kg
400 46 0/10
200 154 4/10 ) toxic dose
100 254 8/10 highest non-toxic dose
50254 7/10
BCNU mg/kg
24 255 10/10 highest non-toxic dose
12 139 0/10
: 3. B 16 Melanocarcinoma i.p. The drugs are administered
in one daily dose from day 1
to 9 after implantation of the tumor.
Lu 16-035 T/C %Survivors/total
;
mg/kg on daY 60
358 4/10
12.5 2~0 1/10
6.25 164 0/10
Lu 16-022 mg/kq
100 153 0/10
50 332 2/10
- 30 25 237 1/10
1~4Z183 ~
13
There is no direct comparison with other nitrosoureas but,
~t is reported in the literature that BCNU, CCNU and Me-CCNU
can give at most a doubling of the survival time.
The compound N-(2-chloroethyl)-N'-~2,2-diphenyl-5-(dimethyl-
amino)pentyl)-N-nitrosourea (Lu 15-132 for short) has so far
only been tested against P 388 lymphocytic leukemia in mice.
It has shown about the same activity and toxicity as Lu 16-035.
From these results it is seen, that Lu 16-035 and Lu 16-022.
have better effects than BCNU on some tumors, and that they
show a more favorable therapeutic index.
The compounds of Formula I and the pharmaceutically acceptable
acid addition salts thereof may be a&inistered to animals
such as dogs, cats, horses, sheep or the like, including
human beings, both orally and parenterally, and may be used
for example in the form of tablets, capsules, powders, syrups
or in the form of the usual sterile solutions for injection.
Most conveniently the compounds of Formula I are administered
orally in unit dosage form such as tablets or capsules, each
dosage unit containing a compound of Formula I or a pharma-
ceutically acceptable salt thereof in an amount of from about1 to about 100 mg, most preferably, however, from about 10 to
50 calculated as the free amine.
The compounds of Fo~mula I are usually administered in intervals
of from four to ten weeks. The exact individual dosages in a
particular case will, of course, be determined according to
established medical principles under the direction of a physician.
The dosage range for such cytostatic drugs is usually given as
we~ght per squaremeter of body area and normally falls within
~rom about 10 to about 200 milligrams per squaremeter of body
area.
,9,~ ,,
.n~ ---1 3
~ ~14;~1~93 14
When preparing tablets, the active ingredient is for the most
part mixed with ordinary tablet adjuvants such as corn starch,
potato starch, talcum, magnesium stearate, gelatine, lactose,
gums, or the like.
As previously stated, when isolating the compounds of Formula I
in the for~ of an acid addition salt the acid is preferably
selected so as to contain an anion which is non-toxic and
pharmacologically acceptable, at least in usual therapeutic
doses. Representative salts which are included in this
preferred group are the hydrochlorides, hydrobromides,
sulphates, acetates, phosphates, nitrates, methanesulphonates,
ethane-sulphonates, lactates, citrates, tartrates or bi-
tartrates, embonates and maleates of the amines of Formula I.
Other acids are likewise suitable and may be employed if
desired. For example: Fumaric, benzoic, ascorbic, succinic,
salicylic, bismethylenesalicylic, propionic, gluconic, malic,
malonic, mandelic, c~nnamic,citraconic, stearic, palmitic,
; itaconic, glycolic, benzenesulphonic, and sulphamic acids
may also be employed as acid addition saltforming acids.
When it is desired to isolate a compound of the invention
in the form of the free base, this may be done according to
conventional procedure as by dissolving the isolated or un-
isolated salt in water, treating with a suitable alkaline
material, extracting the liberated free base with a suitable
organic solvent drying the extract and evaporating to dryness
or fractionally distilling to effect isolation of the free
basic amine.
The invention also comprises a method for the alleviation,
palliation, mitigation or inhibition of the manifestations
of certain physiological-psychological abnormalies of animals
by administering to a living animal body, including human beings,
an adequate quantity of a compound of Formula I or a non-toxic
acid addition salt thereof. An adequate quantity would be
from about 5 mg to about 100 mg per squaremeter of body area
in each unit dosage, and from about 15 milligrams to about
300 milligrams/squaremeter of body area every fourth to sixth
week.
114Z183 15
It is to be understood that the invention is not limited to
the exact details of operation or exact compound or compositions
shown and described, as obvious medifications and equivalents
will be apparent to one skilled in the art.
;. .
