Note: Descriptions are shown in the official language in which they were submitted.
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Title
ANTITUMOR COMPOSITIONS AND METHODS OF TREATMENT
Background of the Invention
In recent years fundamental advances have been made in
the development of chemical agents and regimens of therapy to combat
neoplastic diseases. Despite these continuing advances, cancers continue
to exact intolerable levels of human pain and suffering. The need for new
and better methods of treating neoplasms and leukemias continues to fuel
efforts to find new classes of antitumor compounds, especially in the area
of inoperable or metastatic solid tumors, such as the various forms of lung
cancer. Of the one million new cases of cancer diagnosed in the United
States each year, more than 90% represent non-hematopoetic tumors,
where improvements in five-year survival rates have been modest, at best.
B.E. Henderson, stet al., Science, 254:113Z-1137 (1991).
The recent avalanche of information regarding the basic
biological processes involved in neoplasms has led to a deeper
understanding of the heterogeneity of tumors. Ongoing work has led to
the realization that individual tumors may contain many subpopulations
of neoplastic cells that differ in crucial characteristics, such as karyotype,
morphology, immunogenicity, growth rate, cap acity to metastasize, and
response to antineoplastic agents.
It is because of this extreme heterogeneity among
populations of neoplastic cells that new chemotherapeutic agents should
have a wide spectrum of activity and a large therapeutic index. In
addition, such agents must be chemically stable and compatible with
other agents. It is also important that any chemotherapeutic regimen be
as convenient and painless as possible to the patient.
This invention reports a series of novel sulfonylureas that
are useful in the treatment of solid tumors. These compounds are orally
active -- which, of course, results in less trauma to the patient -- and are
relatively non-toxic. These compounds also have an excellent therapeutic
index. The compounds and their formulations are novel.
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Many sulfonylureas are known in the art. Certain of these
compounds are known to have hypoglycemic activities, and have been
used medicinally as such agents. In addition, some suifonylureas have
been taught to have herbicidal and antimycotic activities. General
reviews of compounds of this structural type are taught by Kurzer,
Chemical Revie~~, 50:1 (1952) and C.R. Kahn and Y. Shechter, Goodman
and Gilman's. The Pharmacological Basis of Therapeutics, (Gilman, et al.)
8th ed. 1990) 1484-1487.
Some diarylsulfonylureas have been reported as being active
ZO antitumor agents. g,gs, U.S. Patent 5,169,860, of F. Mohamadi and M.
Spees, issued December 8, 1992; U.S. Patent 4,845,128 of Harper, etet al.,
issued July 4, 1989; U.S. Patent 5,110,830 of Harper, etet al.) issued May 5)
1992; U.S. Patent 5,116,874 of G.A. Poore, issued May 26, 1992; U.S.
Patent 5,216,026) of J. Howbert, issued June 1, 1993; U.S. Patent
5,216,027, of J.E. Ray) etet al., issued June 1, 1993; U.S. Patent 5,260,338,
of R.W. Harper, etet al., issued November 9, 1993; U.S. Patent 5,594,028, of
R.W. Harper, etet al., issued January i4) 1997; U.S. Patent 5,302,724) of
J.J. Howbert, etet al., issued April 12, 1994; U.S. Patent 5,2?0,329, of W.L.
Scott, et al., issued December 14, 1993; U.S. Patent 5,234,955, of J.E. Ray,
etet al.) issued August 10, 1993; U.S. Patent 5,354,778, of J.E. Ray, etet
al.,
issued October 11, 1994; U.S. Patent 5,258,406, of J.E. Toth, etet al., issued
November 2, 1993; U.S. Patent 5,2f 2,440, of W.J. Ehlhardt, etet al., issued
November 16, 1993; U.S. Patent 5,254,582, of G.B. Boder, etet al., issued
October 19, 1993; U.S. Patent 5,565,494, of G.B. Grindey, etet al., issued
October 15, 1996; U.S. Patent 5,387,681, of W.D. Miller, et al., issued
February 7, 1995; and U.S. Patent 5,529,999, of J.E. Ray, et al., issued
June 25, 1996; the entirety of all of which are herein incorporated by
reference.
SummarLof the Invention
This invention provides novel compounds of Formula I
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O O . ~ Rs
Ria \\ // O
\\ SwN~N \
R2
H H
R1
wherein:
R1 is C2-Cg alkenyl;
Rla is hydrogen or hydroxy; and
RZ and R3 are independently selected from the group
consisting of hydrogen, halo, C 1-Cg alkyl, and trifluoromethyl, provided
that no more than one of R2 and R3 can be hydrogen;
or a pharmaceutically acceptable salt, solvate, or prodrug
thereof.
This invention also provides a method of treating a
susceptible neoplasm in a mammal which comprises administering to a
mammal in need of said treatment an effective amount for treating a
susceptible neoplasm of a compound of Formula I.
In addition, this invention provides pharmaceutical
formulations comprising an effective amount for treating susceptible
neoplasms of a compound of Formula I, or a salt, solvate or prodrug
thereof, in combination with a suitable pharmaceutical carrier, diluent, or
excipient. These formulations are useful in the treatment of mammals
suffering from susceptible neoplasms.
retailed T,escrintion and Preferred Embodiments
As used herein, the term "halo" refers to ffuoro, chloro,
bromo, and iodo. The term "C 1-Cg alkyl" refers to straight or branched,
monovalent, saturated aliphatic chains of 1 to fi carbon atoms and
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includes, but is not limited to, methyl, ethyl, propyl, isopropyl, butyl,
isobutyl, t-butyl, pentyl, isopentyl, and hexyl.
The term "C2-Cg alkenyl" as used herein represents a
straight or branched, monovaient, unsaturated aliphatic chain having
from two to eight carbon atoms. Typical C2-Cg aikenyl groups include
ethenyl (also known as vinyl), 1-methylethenyl, 1-methyl-1-propenyl, 1-
butenyl, 1-hexenyl, 2-methyl-2-propenyl, 1-propenyl, 2-propenyl, 1-
butenyl, 2-pentenyl, and the like.
Preferred methods of treatment employ compounds of
Formula I in which R1 is ethenyl or propenyl; and R2 and R3 are
independently selected from the group consisting of hydrogen, chloro,
fluoro, bromo, iodo, methyl, ethyl, and triffuoromethyl.
The compounds of Formulas I are generally referred to as
derivatives of N-[[(substituted phenyl)amino]carbonyl]-
(alkenylbenzene)sulfonamides. Alternatively, the compounds can be
referred to as 1-(substituted phenyl)-3-(substituted phenylsulfonyl)ureas
or N- and N'-substituted sulfonylureas.
The compounds of Formulas I can be prepared by methods
known in the literature. Generally, these methods involve either the
reaction of a sulfonamide with an isocyanate, a reaction of a
sulfonylisocyanate with an appropriately substituted aniline, or a reaction
of a sulfonylcarbamate with an appropriately-substituted aniline.
A preferred process for preparing a compound of Formula I
comprises reacting a sulfonylisocyanate of Formula II
R1
O
S- NCO
I!
O
II
with an aniline derivative of Formula III
R2
H2N ~ ~ R3
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III
where R1, R2, and R3 are the same as previously defined.
The reaction between compounds II and III is usually
performed using equimolar amounts of the two reactants, although other
ratios are operative. The reaction is preferably carried out in a solvent
which is nonreactive under the reaction conditions such as benzene,
toluene, acetonitrile, diethyl ether, tetrahydrofuran, dioxane, methylene
chloride, or acetone.
The reaction can be carried out at temperatures from about
O~C up to about 100~C. At the preferred temperature range of from about
20C to about 30C) the reaction produces a strong exotherm and the
reaction is usually complete within one hour. The product thus obtained
is recovered by filtration and can be purified, if desired, by any number of
methods known to those skilled in the art, such as chromatography or
crystallization.
An alternative preferred process for preparing a compound of
Formula I comprises reacting an appropriately substituted sulfonamide of
Formula IV
R1
O
i - NH2
O
IV
with an isocyanate of Formula V
R2
OCN ~ ~ R3
V
to provide the corresponding compound of Formula I.
The reaction is generally performed in a mixture of water
and a water-miscible, non-reactive solvent such as tetrahydrofuran or
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acetone in the presence of a base such as sodium hydroxide, potassium
hydroxide, lithium hydroxide, sodium methoxide, sodium hydride and the
Iike. Generally, an equimolar or slight molar excess of V is employed,
although other ratios are operative. Usually, the amount of base used is
approximately equimolar to the amount of IV. The reaction is generally
carried out from about 0~C up to about 100~C. At the preferred
temperature of about 20~C to about 30~C, the reaction is usually complete
within about three hours.
A preferred process for preparing a compound of Formula I
involves reacting a sulfonamide of Formula IV with an alkyl haloformate
of the formula XCOOR4~ where X is bromo or chloro and R4 is C 1-Cg alkyl,
to provide the carbamate of Formula VI and then reacting it with an
aniline derivative of Formula III to provide the corresponding product of
Formula I.
R1 0
IV + XCOOR4 ~ ~ (I III
II ~ COOR4 ~ I
O
VI
The transformation of IV into VI is usually accomplished in a non-reactive
solvent, such as acetone or methyl ethyl ketone, in the presence of an acid
scavenger, such as an alkali metal carbonate, for example potassium
carbonate. A molar excess of the haloformate is usually added, although
other ratios are operative. The reaction mixture is heated to a
temperature from about 30~C up to the reflux temperature of the mixture
for a period of about 1-6 hours to provide the desired intermediate VI.
Intermediate carbamate VI and the substituted aniline III are then
heated together in an inert high-boiling solvent, such as dioxane, toluene,
or diglyme, at temperatures from about 50~C up to the reffux temperature
of the mixture to provide the desired product of Formula I.
The carbamate of Formula VI can also be synthesized by the
procedure described by Atkins and Burgess. G. Atkins and E. Burgess,
Journal of the American Chemical Society, 94:6135 (1972). In this process
triethylamine and a substituted aniline are mixed in the presence of a
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solvent such as benzene. To this mixture a sulfamoyl chloride is added to
produce the carbamate of Formula VI.
Intermediates II, III) IV, V, and VI, and any other reagents
required for these methods of preparation are commercially available, are
known in the literature, or can be prepared by methods known in the art.
This invention encompasses the pharmaceutically acceptable
salts of the compounds defined by Formula I. Although generally neutral,
a compound of this invention can possess a sufficiently acidic functional
group, and accordingly react with any of a number of organic or inorganic
bases to form a pharmaceutically acceptable salt.
The term "pharmaceutically acceptable salt" as used herein,
refers to salts of the compounds of Formula I which are substantially non-
toxic to living organisms. Typical pharmaceutically acceptable salts
include those salts prepared by reaction of the compounds of the present
invention with a pharmaceutically acceptable organic or inorganic base.
Such salts are known as base addition salts.
Base addition salts include those derived from inorganic
bases, such as ammonium or alkali or alkaline earth metal hydroxides,
carbonates, bicarbonates, and the like. Such bases useful in preparing
the salts of this invention thus include sodium hydroxide, potassium
hydroxide, ammonium hydroxide, potassium carbonate) sodium
carbonate, sodium bicarbonate, potassium bicarbonate, calcium
hydroxide, calcium carbonate, and the like. The potassium and sodium
salt forms are particularly preferred. Organic bases can also be used,
including primary, secondary, and tertiary alkyl amines such as
methylamine, triethylamine, and the like.
It should be recognized that the particular counterion
forming a part of any salt of this invention is not of a critical nature, so
long as the salt as a whole is pharmacologically acceptable and as long as
the counterion does not contribute undesired qualities to the salt as a
whole.
This invention further embraces the pharmaceutically
acceptable solvates of the compounds of Formulas I. The Formula I
compounds can combine with solvents such as water, methanol, ethanol
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and acetonitrile to form pharmaceutically acceptable solvates such as the
corresponding hydrate, methanolate, ethanolate and acetonitrilate.
This invention also encompasses the pharmaceutically
acceptable prodrugs of the compounds of Formula I. A prodrug is a drug
which has been chemically modified and may be biologically inactive at its
site of action, but which may be degraded or modified by one or more
enzymatic or other inin vivo processes to the parent bioactive form. This
prodrug should have a different pharmacokinetic profile than the parent,
enabling easier absorption across the mucosal epithelium, better salt
formation or solubility, or improved systemic stability (an increase in
plasma half life, for example).
Typically, such chemical modifications include:
1) ester or amide derivatives which may be cleaved by
esterases or lipases;
2) peptides which may be recognized by specific or
nonspecific proteases; or
3) derivatives that accumulate at a site of action through
membrane selection of a prodrug form or a modified prodrug form;
or any combination of 1 to 3, sugra. Conventional procedures for the
selection and preparation of suitable prodrug derivatives are described,
for example, in H, Bundgaard, Design of Prodru~s, (1985).
The terms and abbreviations used in the instant examples
have their normal meanings unless otherwise designated. For example
"_C" refers to degrees Celsius; "N" refers to normal or normality; "mmole"
refers to milli.mole; "g" refers to gram or grams; "ml" means milli)iter or
milliliters; "M" refers to molar or molarity; "FDMS" refers to field
desorption mass spectrometry; "m.p." refers to melting point; and "NMR"
refers to nuclear magnetic resonance.
The following examples further illustrate the preparation of
the compounds of Formula I. These examples are illustrative only and
are not intended to limit the scope of the invention in any way.
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Examp~'~e 1
Preparation of N-([(4-chlorophenyl)amino]carbonyl]-4-
(ethenyl)benzenesulfonamide.
C1
o~ ~i o /
I
N N
H H
HaC w ~ /
To a solution of 4-vinylbenzenesulfonamide, prepared
essentially as described in R.H. Wiley, etet al., Journal of the American
Chemica~j Society) 78:2169 (1956), (5.1 g) 28 mmol) in 1N aqueous sodium
hydroxide solution (28 ml) and acetone (14 ml) was added, dropwise, a
solution of 4-chlorophenylisocyan ate (4.3 g, 2 8 mmoles) in acetone ( 14 ml)
over 10 minutes. Two hours later the mixture was filtered and the filtrate
treated with 1N hydrochloric acid {28 ml). The resulting solid was
collected by filtration and rinsed with water (100 ml). The crude product
was purified by dissolving in 100 ml water containing 40m1 1N sodium
hydroxide, followed by filtration of insoluble material and neutrailization
with 40 ml 1N hydrochloric acid. After filtration and washing (200 ml of
water), vacuum drying gave 4.9 g (52%) of the purified title sulfonylurea.
Analysis of the product gave the following results: mp=182-
184~C; Rf(911, CHCI3lMeOH)=0.30; 1H NMR (300 MHz, d6-DMSO) b 5.45
(d, 1H, J=1o.9 Hz, CI-~I , 5.9s (d, 1H, J=17.6 Hz, Cue, 6.83(dd, 1H,
J=10.9,17.6 Hz, CF-~) 7.2-7.4 (m, 4H, Ar-,~, 7.68 (d, 2H, J=8.3 Hz, Ar-~,
7.89 (d, 2H, J=8.3 Hz, Ar-~, 9.0 (s, 1H) exchanges with D20, N -~I and
10.9 (bs, 1H, exchanges with D20, S02N~J; IR(KBr) 3360) 1710, 1604,
1542, 1462, 1340, 1161, 1034 and 927 cm-1; W(EtOH) ~,m~(s) 251.2
{33271) and 204.4 (36099) nm; FDMS (MeOH) mle 336, 338(M+).
Analysis for C 15H 13C1N203S:
Theory: C, 53.49; H, 3.89; N, 8.32.
Found: C, 53.54; H, 3.96; N, 8.19.
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Exams
Preparation of N-[[(3,4-dichlorophenyl)amino]carbonyl]-4-
(ethenyl)benzenesulfonamide.
//~~ Cl
O
\ S~ ~ \
H H C1
The procedure of Example 1 was followed, using 4-
vinylbenzenesulfonamide (IO g, 55 mmoles), IN sodium hydroxide (55 ml)
and 3,4-dichlorophenylisocyanate (97%, 11 g, 55 mmoles). The crude
product was purified by stirring in ethanol (50 ml) for 30 minutes,
followed by filtration and vacuum drying to give 8.4 g (41%) of the title
sulfonylurea.
Analysis of the product gave the following results:
mp=179~C; Rf(911, CHC131MeOH)=0.24 ; 1H NMR (300 MHz, dg-DMSO) 8
5.45 (d, 1H, J=10.9 Hz, Cue, 5.98 (d, 1H, J=17.6 Hz) CI-~I , 6.83(dd, 1H,
J=10.9, 17.6 Hz, Cue, 7.25 (m, 1H, Ar-H>, 7.43 (d, 1H, J=8.7 Hz, Ar-I-~I ,
7.s-7.7 (m, 3H, Ar-Hue, 7.88 {d, 2H, J=8.7 Hz, Ar-I-~, 9.1 (s, 1H, exchanges
with D20, NI-~i and 11.0 (ins, 1H, exchanges with D20, S02NH~; IR(KBr)
3347, 3250, 1710, 1589, 1521, 1464, 1338, 1161, 1040 and 843 cm' I;
LTV(EtOH) ~.m~(s) 254.0 (33940) and 209.6 (41966) nm; FDMS (MeOH)
m/e 370, 3?2, 374(M+).
Analysis for C 15H 12C12N203S:
Theory: C, 48.53; H) 3.26; N, 7.55.
Found: C, 48.34; H, 3.29; N, 7.39.
xample 3
Preparation of N-[[(4-chlorophenyl)amino]carbonyl]-3-
(ethenyl)benzenesulfonamide.
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C1
O O /
/ Sw
H2C / \ H H \
/
The 3-vinylbenzenesulfonamide was prepared by reacting a
solution of 3-bromobenzenesulfonamide (24 g, 100 mmoles) with
vinyltributyltin (97%, 35 ml, 116 mmoles) in the presence of
tetrakis(triphenyiphosphine)palladium(0) (2.3 g, 2 mmoles) in toluene
(200 ml) and heated under nitrogen at reflux for 90 minutes. After
cooling and filtering through CELITE~) the reaction mixture was
evaporated to yield 50 g of yellow solid. Chromatography (silica gel, 100%
hexanes to 40% ethyl acetatelhexane) provided 5.2g (28%) of the 3-
vinylbenzenesulfonamide. Recrystallization from methanol gave an
analytical sample.
Analysis of the product gave the following results: mp=131-
132~C; Rf(l/1, EtOAc/hexane)=0.48 ; 1H NMR (300 MHz, d6-DMSO) 8
5.38 (d, 1H) J=10.9 Hz, Cue, 5.89 (d, 1H, J=17.6 Hz, Cue, 6.83(dd, 1H,
J=10.9,17.6 Hz, C -~i , 7.35 (s, 2H, S02N~) 7.50 (m, 1H, Ar-I-~I , 7.7 (m,
2H, Ar-~ and 7.89 (s, 1H, Ar- -~I ; IR(KBr) 3333, 3246, 1557) 1328, 1158,
and 890 cm-1; UV(EtOH} 7~max(E) 297.8 (452), 288.6 (730), 280.8 (763),
248.2 (11423) and 214.8 (25659) nm; FDMS (MeOH) mle 183(M+).
Analysis for CgHgN02S:
Theory: C, 49.62; H, 4.95; N, 7.64.
Found: C, 49.90; H, 4.61; N, 7.32.
The title compound was prepared essentially as described in
Example I, using the 3-vinylbenzenesulfonamide (3.0 g, 16.4 mmoles)
prepared su ra) 4-chlorophenylisocyanate (2.6 g) l6.6 mmoles) and 1N
sodium hydroxide (16.4 ml). The crude product was chromatographed
(silica gel, I-5% methanol in methylene chloride) to give 2.7 g (49%) of N-
[[(4-chlorophenyl)amino]carbonyl]-3-(ethenyl)benzenesulfonamide.
Analysis of the product gave the following results: mp=153-
154~C; Rf(9I1, CHCI3IMeOH)=0.15; 1H NMR (300 MHz, d6-DMSO) 8 5.38
(d, 1H, J=10.9 Hz, CSI , 5.89 (d, 1H, J=17.6 Hz, Cue, 6.83(dd, 1H,
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J=10.9,17.6 Hz, C -~I , 7.25-7.40 (m, 4H, Ar-~i , 7.58 (m, 1H, Ar-~i , 7.78-
7.85 (m, 2H, Ar-H~, 7.97 (s, 1H, Ar-~, 9.1 (s, 1H, exchanges with D20,
NI-~ and 10.8 (bs, 1H, exchanges with D20, S02NI-~i ; IR(KBr) 3329, 3239,
1705, 1598, 1534, I456, I337, 1158, 1038 and 927 cm-I; W(EtOH)
~.m~(s) 247.8 (32235) and 204.2 (35716) nm; FDMS (MeOH) m/e 336, 338
(M+). Analysis for C15H13C1N203S:
Theory: C, 53.49; H, 3.89; N, 8.32.
Found: C, 53.45; H, 3.98; N, 8.20.
Pre~ aration
Preparation of N-ethoxycarbonyl-4-hydroxy-3-(prop-2-en-I-
yl)phenylsulfonamide
O O
H2C g
\ N~O~CH3
H
HO
To a solution of 3-allyl-4-hydroxybenzenesulfonamide (2 g,
9.4 mM) [prepared as described in Patent Cooperation Treaty Publication
WO 96I09818, published April 4, 1996) in l00 ml methyl ethyl ketone was
added potassium carbonate (1.4 g, 10 mM) and the mixture heated under
nitrogen to reflux. After 10 minutes, ethyl chloroformate (1.8 ml, 18.8
mM, 2 equivalents) was added dropwise and the mixture refluxed for 3
hours. The cooled reaction was acidified with 1N hydrochloric acid (50
mI) and the layers separated. An additional wash with water (1 x 50 ml)
and brine (1 x 24 ml), followed by drying over sodium sulfate and
evaporation gave 3.3 g of the di-acylated product. The crude oil was
dissolved in water (90 ml), IN sodium hydroxide (29 ml) added and the
mixture heated to refl.ux 2 hours. After cooling in an ice-bath, 1N
hydrochloric acid (30 ml) was added and the resulting solid collected and
dried to 1:87 g. Chromatography (silica gel, 100% methylene chloride to
5% methanol/methylene chloride) gave the product as a white solid, 1.3 g
(49%).
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Analysis of the title compound gave the following results:
Rf (I/9 MeOHICHC13) = 0.29;
1H NMR (300 MHz, DMSO-dg) b 1.05 (t, 3H, J=6.9 Hz, OCH2C~), 3.30 (d)
2H, J=6.7Hz, CHZ=CHC,~), 3.95 (q, 2H, J=6.9Hz, OCH~C~), 5.05 (s, 1H,
C~=CHCH~, 5.05 (d, 1H, J= 4Hz, C -~I =CHCHZ}, 5.85-5.94 (m, 1H,
CHZ=CHCHz), 6. 91 (d, 1H, J=8.4Hz, Ar-H)) 7.50-7.60 (m, 2H, Ar-H),
10.6(bs, 1H, exchanges with D20, OH), and 11.63(bs, 1H, exchanges with
DZO, NH);
IR(KBr) 3409, 3212, 1728, 1592, 1488, 1425, 1366, I348, 1290, 1239,
1159, 1129, 835 and 772 cm-1;
UV(EtOH} ~,max(E) 244.5 (I3070) and 207.0 (31769) nm;
FDMS (MeOH) m/e 285 (M~.
Analysis for C12H15N05S:
Theory: C, 50.52; H, 5.30; N) 4.91.
Found: C, 50.39; H, 5.20; N, 4.66.
Preparation of N-[[[3,4-dichlorophenyl]amino]carbonyl]-4-hydroxy-3-
(prop-2-en-1-yl)phenyisulfonamide
O 0 /~ Cl
HaC \S// I
H H C1
HO
A solution of N-ethoxycarbonyl-4-hydroxy-3-(prop-2-en-1-
yl}phenylsulfonamide (500 mg, 1.75 mM) and 3,4-dichloroaniline (340 mg,
2.06 mM, 1.2 equivalents) was prepared in toluene (25 ml) and heated to
reflux under nitrogen, removing the toluene/ethanol azeotrope by use of a
Dean-Stark trap. After 4.5 hour and removal of about 10 ml azeotrope,
the solution was cooled in an ice-bath and the resulting precipitate
collected and rinsed with cold toluene (10 ml). Vacuum dry to a white
solid, 536 mg (76%).
s
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Analysis of the title compound gave the following results:
Rf {1l9 MeOHlCHCI3) = 0.19;
'H NMR (300MHz, DMSO-d6)8 3.31 (d, 2H, J=6.7Hz, CHZ=CHC -~i )) 5.01
(s, 1H, C~=CHCHZ), 5.06 {d, 1H, J= 7.9Hz, C -~I =CHCHZ), 5.85-5.94 (m,
1H) CHZ=CHCHZ), 6. 91 (d, 1H, J=8.4Hz, Ar-H), 7.22 (m, 1H, Ar-H), 7.45
(d, 1H, J=8.8Hz, Ar-H)) 7.50-7.65 (m, 3H, Ar-H), 9.03 (bs, 1H, exchanges
with D20, CONH), 10.6(bs, 1H, exchanges with D20, OH), and 10.8(bs,
1H, exchanges with D20, NH); ;
IR(KBr) 3383, 3206, 1712, 1700, 1591, 1521, 1460, 1432, 1394, 1339,
1279, 1239, 1152, 1 Z23, 1056, 864 and 706cm-';
FDMS (MeOH) mle 400,402 (M~, M++2).
Analysis for C 1gH 14C12N204S:
Theory: C, 47.89; H, 3.52; N, 6.98.
Found: C, 47.78; H, 3.61; N, 6.72.
The compounds of Formula I have been shown to be active
against human tumors in ' r . The in vitro data were obtained using
CCRF-CEM cells, a human leukemia cell line. Foley etet al., er, 18:522
(1965). These cells were grown using standard techniques. See, e.g., G.B.
Grindey, et , journal of Molecular Pharmacoloev, 16:601 {1979). Dose-
response curves were generated for various compounds to determine the
concentration required for 50% inhibition of growth (ICSp). Cluster plates
were prepared in duplicate with the compound at various concentrations.
Test compounds were dissolved initially in DMSO at a concentration of 4
mg/ml and further diluted with solvent to the desired concentration. Cells
in Roswell Park Memorial Institute 1640 media supplemented with 10%
dialyzed fetal bovine serum, and 25 mM HEPES buffer were added to the
well at a final concentration of 4.8 X 104 cellslwell in a total volume of 2.0
ml. After 72 hours of incubation (95% air, 5% C02), cell numbers were
determined on a ZBI Coulter counter. Cell number for indicated controls
at the end of incubation was usually (4-6) X 105 cellslwell. Cell viability
was also measured by staining with 3-[4,5-dimethylthiazol-2-yl]-2,5-
diphenyl-tetrazolium bromide (MTT) using standard techniques. R.I.
Freshney, Culture of Animal Cells: A Manual of Basic Technigue, 253-254
(2d ed. 1987).
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Table I, infra, shows the results of one such in vitro
screening panel. Column 1 refers to the example number of the compound
tested; Column 2 depicts the in ' r cytoxicity against CCRF-CEM cells
by relating the concentration of the test compound required for 50%
inhibition of growth (IC5p) of the cells in the well.
Table I
Activity of the Compounds of Formula I Against
Tumor Cells I_n Vitro
Example (CCRF-CEM)
No. ICSO ~.glml
1 19.8
2 9.3
3 14.4
4
The compounds of Formula I have also been shown to be
active against transplanted human tumors in vivo. To demonstrate the
anti-tumor activity of the compounds of Formula I, these compounds were
tested in mice bearing different allograft and xenograft tumors.
One of the tumor models used for showing the anti-neoplastic
activity of the sulfonylureas of this invention was the human colon
xenograft HXGC3. J.A. Houghton and D.M. Taylor, British Journal of
~, 37:213-223 (1978). This tumor was obtained from St. Jude's
Children's Research Hospital and has been widely used as a human
tumor model.
A second tumor model employed C3H mice bearing the
widely used allograft 6C3HED lymphosarcoma, also known as the
Gardner lymphosarcoma (GLS). The 6C3HED lymphosarcoma was
obtained from the Division of Cancer Treatment, National Cancer
Institute, Tumor Bank, maintained at E. G. and G. Mason Research
(Worcester, Massachusetts).
First passage tumors were stored in liquid nitrogen, using
standard techniques. The transplanted tumor was reestablished from the
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Tumor Bank every six months or as needed. The tumor was maintained
by serial passage twice weekly in the host mice.
In the procedures utilized here) the tumor was removed from
passage animals and minced into 1- to 3-mm cubic fragments using sterile
techniques. Tumor pieces were checked for sterility using both Antibiotic
Medium 1 and Brain Heart Infusion (Difco, Detroit, Michigan). The
xenograft tumor pieces were implanted into the recipient CD 1 Nu/Nu
mice subcutaneously in an axillary site by trochar. The allograft 6C3HED
tumor pieces were implanted into the recipient C3H mice in an analogous
fashion.
Drug therapy on the appropriate schedule was initiated
seven days after tumor implantation for the HXGC3 xenograft and the
day after tumor implantation for the 6C3HED allograft. The compound
being tested was mixed with 2.5% Emulphor EL620 from GAF
Corporation (1:40 dilution in 0.9% saline). The total dosage volume for
each administration was 0.5 ml. All animals were weighed at the
beginning and end of administration of the subject compounds. Food and
water were provided ad libitum.
Each control group and each dosage level of the treated
groups consisted of 9 or 10 mice selected at random from the pool of
implanted animals. The formulations were administered orally by gavage
with the use of an 18-gauge needle. Compounds were dosed daily for 10
days for the studies using the human tumor xenografts and 8 days for the
studies using the allograft.
The xenograft tumor was measured five days after treatment
ended with two dimensional measurements (width and length) of the
tumor taken using digital electronic calipers interfaced to a
microcomputer. J.F. Worzalla, et al.) Investigational New Drues, 8:241-
251 (1990). The allograft was measured in a like manner the day after
the dosing schedule ended. Tumor weights were calculated from these
measurements using the following formula:
Tumor weight (mg) = tumor length (mm) x jtumor width ~mm)12
2
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At least one control group of an equal number of mice was treated with
the same volume of 2.5% Emulphor only. The percent inhibition is
determined by subtracting the ratio of the mean tumor size of the test
group relative to the control group from one and multiplying the result by
100.
The results of several experiments in mice bearing the
HXGC3 human colon adenocarcinoma and the 6C3HED lymphosarcorna
when the Formula I compounds were administered orally are provided in
Table II. In the table, Column 1 refers to the example number of the
compound tested; Column 2 describes the particular human tumor
xenograft or mouse allograft being studied; Column 3 gives the dosage
level of the compound of Formula I in milligrams per kilogram of body
weight; Column 4 describes the percent inhibition of tumor growth; and
Column 5 tallies the number of mice which died during the course of the
experiment relative to the total number of animals in the group.
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Table iI
Activity of the Compounds of Formula I Against
Allograft and Xenograft Tumors In Vivo
Example Dosage Percent
No. Tumor (mg/k~) Inhibition Toxic/Total
1 6C3HE 300 99 1I10
D
150 99 0l10
6C3HE 1200 Toxic 10I10
D
600 100 2I10
300 10Q 1I10
150 97 OI10
75 66 O/10
3 7.5 34 0110
2 HXGC3 300 97 1I7
150 93 017
3 6C3HE 300 83 O/10
D
150 60 OI10
The compounds of Formula I are usually administered in the
form of pharmaceutical compositions. These compositions can be
administered by a variety of routes including oral, rectal, transdermal,
subcutaneous, intravenous, intramuscular, and intranasal. Such
compositions are useful in treating solid tumors including carcinomas
such as ovarian, non-small cell lung, gastric, pancreatic, prostate, renal
cell, breast, colorectal, small cell lung, melanoma, and head and neck; and
sarcomas such as Kaposi's sarcoma and rhabdomyosarcoma.
The Formula I compounds are preferably administered in the
form of oral pharmaceutical compositions. Such compositions are
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prepared in a manner well known in the pharmaceutical axt and comprise
at least one active compound.
In another aspect, the present invention also includes novel
pharmaceutical compositions which contain, as the active ingredient) the
compounds of Formula I associated with pharmaceutically acceptable
carriers. In making the compositions of the present invention the active
ingredient is usually mixed with an excipient, diluted by an excipient or
enclosed within such a carrier which can be in the form of a capsule,
sachet, paper or other container. When the excipient serves as a diluent,
it can be a solid, semi-solid, or liquid material, which acts as a vehicle,
carrier or medium for the active ingredient. Thus, the compositions can
be in the form of tablets, pills, powders, lozenges, sachets, cachets,
elixirs,
suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a
liquid medium), ointments containing for example up to 10% by weight of
the active compound, soft and hard gelatin capsules, suppositories) sterile
injectable solutions, and sterile packaged powders.
In preparing a formulation, it may be necessary to mill the
active compound to provide the appropriate particle size prior to
combining with the other ingredients. If the active compound is
substantially insoluble, it ordinarily is milled to a particle size of less
than
200 mesh. If the active compound is substantially water soluble, the
particle size is normally adjusted by milling to provide a substantially
uniform distribution in the formulation, e.g. about 40 mesh.
Some examples of suitable excipients include lactose,
dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium
phosphate, alginates, tragacanth, gelatin, calcium silicate,
microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup,
and methyl cellulose. The formulations can additionally include:
lubricating agents such as talc, magnesium stearate, and mineral oil;
wetting agents; emulsifying and suspending agents; preserving agents
such as methyl- and propylhydroxybenzoates; sweetening agents; and
flavoring agents. The compositions of the invention can be formulated so
as to provide quick, sustained or delayed release of the active ingredient
after administration to the patient by employing procedures known in the
art.
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The compositions are preferably formulated in a unit dosage
form, each dosage containing from about 5 to about 500 mg, more usually
about 25 to about 300 mg, of the active ingredient. The term "unit dosage
form" refers to physically discrete units suitable as unitary dosages
dosages for human subjects and other mammals, each unit containing a
predetermined quantity of active material calculated to produce the
desired therapeutic effect, in association with a suitable pharmaceutical
excipient.
The active compounds are effective over a wide dosage range.
For examples, dosages per day normally fall within the range of about 0.5
to about 600 mg/kg of body weight. In the treatment of adult humans, the
range of about 1 to about 50 mg/kg, in single or divided dose, is preferred.
However, it will be understood that the amount of the compound actually
administered will be determined by a physician, in the light of the
relevant circumstances, including the condition to be treated, the chosen
route of administration, the actual compound administered, the age,
weight, and response of the individual patient, and the severity of the
patient's symptoms, and therefore the above dosage ranges are not
intended to limit the scope of the invention in any way.
Typical compositions of this invention are described in the
following examples:
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Formulation Exam 1~1
Hard gelatin capsules containing the following ingredients
are prepared:
Quantity
Ingredient (mglca sule)
Active Ingredient 250.0
Starch 305.0
Magnesium stearate 5.0
The above ingredients are mixed and filled into hard gelatin
capsules in 560 mg quantities.
Formulation Example 2
A tablet formula is prepared using the ingredients below:
Quantity
Ingredient (mg/tablet)
Active Ingredient 250.0
Cellulose, microcrystalline 400.0
Colloidal silicon dioxide 10.0
Stearic acid 5.0
The components are blended and compressed to form tablets,
each weighing 665 mg.
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Formulation Examnl
A dry powder inhaler formulation is prepared containing the
following components:
In~:redient ~ %
Active Ingredient 5
Lactose 95
The active mixture is mixed with the lactose and the mixture
is added to a dry powder inhaling appliance.
Formulation Example 4
Tablets, each containing fi0 mg of active ingredient, are
prepared as follows:
Quantity
Ingredient ~~ne/tablet)
Active Ingredient 60.0 mg
Starch 45.0 mg
Microcrystalline cellulose 35.0 mg
Polyvinylpyrrolidone
(as IO% solution in water) 4.0 mg
Sodium carboxymethyl starch 4.5 mg
Magnesium stearate 0.5 mg
Talc 1.4 mg
Total 150 mg
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The active ingredient, starch and cellulose are passed
through a No. 20 mesh U.S. sieve and mixed thoroughly. The solution of
polyvinylpyrrolidone is mixed with the resultant powders, which are then
passed through a 16 mesh U.S. sieve. The granules so produced are dried
at 50-60C and passed through a 16 mesh U.S. sieve. The sodium
carboxymethyl starch, magnesium stearate, and talc, previously p assed
through a No. 30 mesh U.S. sieve, are then added to the granules which,
after mixing, are compressed on a tablet machine to yield tablets each
weighing 150 mg.
to
Formulation Exams
Capsules, each containing 80 mg of medicament are made as
follows:
Quantity
~,~xedient fmg/ca sule)
Active Ingredient 80.0 mg
Staxch 109.0 mg
Magnesium stearate 1.0 mg
Total 190.0 mg
The active ingredient, cellulose, starch, and magnesium
stearate are blended, passed through a No. 20 mesh U.S. sieve, and filled
into hard gelatin capsules in 190 mg quantities.
n ~ i
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Formulation Example 6
10
Suppositories, each containing 225 mg of active ingredient
are made as follows:
Ingredient Amount
Active Ingredient 225 mg
Saturated fatty acid glycerides to 2,000 mg
The active ingredient is passed through a No. 60 mesh U.S.
sieve and suspended in the saturated fatty acid glycerides previously
melted using the minimum heat necessary. The mixture is then poured
into a suppository mold of nominal 2.0 g capacity and allowed to cool.
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mulation~
Suspensions, each containing 50 mg of medicament per 5.0
ml dose are made as follows:
In a 'ent ount
Active Ingredient 50.0 mg
Xanthan gum 4.0 mg
Sodium carboxymethyl cellulose (11%)
Microcrystalline cellulose (89%) 50.0 mg
Sucrose 1.75 g
Sodium benzoate 10.0 mg
Flavor q.v.
Color q.v.
Purified water to 5.0 m1
The medicament, sucrose and xanthan gum are blended,
passed through a No. 10 mesh U.S. sieve, and then mixed with a
previously made solution of the microcrystalline cellulose and sodium
carboxymethyl cellulose in water. The sodium benzoate, flavor, and color
are diluted with some of the water and added with stirring. Sufficient
water is then added to produce the required volume.
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Formulation Example 8
Capsules, each containing 150 mg of medicament, are made
as follows:
Quantity
I~gre client ~m~lcan sule~
Active Ingredient 150.0 mg
Starch 407.0 mg
Magnesium stearate 3_U m~
Total 560.0 mg
The active ingredient, cellulose, starch, and magnesium
stearate axe blended, passed through a No. 20 mesh U.S. sieve, and filled
into hard gelatin capsules in 560 mg quantities.
