|Title of Invention||
"1-(4-SULFAMYLARYL)-3-TRIFLUOROMETHYL-5-ARYL-2-PYRAZOLINES AS INHIBITORS OF CYCLOOXYGENASE-2"
|Abstract||Compounds of the formula wherein: X is selected from the group consisting of trihalomethyl, C1-C6 alkyl, and a group of formula II: wherein: R3 and R4 are independently selected from the group consisting of hydrogen; halogen; hydroxyl; nitro; C1-C6 alkyl; C1-C6 alkoxy; carboxy; C1-C6 trihaloalkyl; and cyano; Z is selected from the group consisting of substituted and unsubstituted aryl; or a pharmaceutically acceptable salt thereof. The compounds are inhibitors of cyclooxygenase-2 activity. They are useful for treating cyclooxygenase- mediated disorders, including, for example, inflamation, neoplastic disorders and angiogenesis-mediated disorders.|
PYRAZOLINES AS INHIBITORS OF CYCLOOXYGENASE-2
Cross-Reference to Related Application
The benefit of the filing date of U.S. provisional patent
application Ser. No. and 60/139,416, filed June 16,1999 is hereby claimed
pursuant to 35 U.S.C. 119(e). The entire disclosure of the aforesaid
provisional application is incorporated herein by reference.
Field of the Invention
The invention relates generally to anti-inflammatory drugs,
and more particularly to novel compounds which inhibit the activity of
Background of the Invention
The metabolites of arachidonic acid, such as prostaglandins,
lipoxygenases and thromboxane products are produced in a wide variety
of tissues and play a key role in several biological responses.
Prostaglandins mediate both beneficial and undesirable biological
reactions. The production of prostaglandins induces pain, swelling, heat
and redness which are characteristic features of inflammation. The chronic
inflammation associated with prostaglandin production leads to the
breakdown of the injured tissue and angiogenesis. In pathologic chronic
inflammation, normal tissues can be destroyed and the new blood vessel
formation can support growth of abnormal tissue. Prostaglandins are also
important for normal physiological processes in different organs. In the
stomach, prostaglandins protect mucosa from acid. They also regulate
blood flow and salt-water balance in the kidney. Prostaglandins are also
important in platelets aggregation and participate in memory and other
Prostaglandins are produced from cell membrane
phospholipids by a cascade of enzymes. The enzymatic activities involve
release of arachidonic acid from the cell membrane by phospholipase A2,
followed by the conversion of arachidonic acid to a common prostaglandin
precursor, PGH2, by cyclooxygenase (also called prostaglandin H
synthase). PGH2 is finally converted to various types of prostaglandins
(PGE,, PGE2, PGI2 or prostacyclin, PGF^ and thromboxane) by cellspecific
Aspirin, and other nonsteroidal anti-inflammatory drugs
(NSAIDs) block the formation of prostaglandins by inhibiting
cyclooxygenase activity. They have analgesic, antipyretic and antiinflammatory
activities. However, chronic treatment with the available
NSAIDs often leads to disruption of beneficial prostaglandin-mediated
processes. The side effects associated with constant usage of NSAIDs
include gastrointestinal (Gl) irritation and formation of life-threatening Gl
A dramatic advance in the field of inflammation research
came with discovery of multiple enzymes for each step of the prostaglandin
synthase cascade. The research suggested that in some situations, such
as inflammation, cyclooxygenase was inducible. The cyclooxygenase
known at the time, cyclooxygenase-1 (COX-1), was clearly non-inducible
or modulated by glucocorticoids. A second, inducible form of
cyclooxygenase known as cyclooxygenase-2 (COX-2) was subsequently
identified and cloned by several groups of investigators. COX-1 is the
constitutive cyclooxygenase isoform and is mainly responsible for the
synthesis of cytoprotective prostaglandins in the Gl tract and the synthesis
of thromboxane which triggers platelet aggregation in blood platelets.
COX-2 is inducible and short lived except in the case of certain tumors
where it is constitutively activated. COX-2 expression is stimulated in
response to endotoxins, cytokines, hormones, growth factors and mitogens.
These observations suggest that COX-1 and COX-2 serve different
physiological and pathophysiological functions. Indeed, it has been
suggested that COX-1 is responsible for endogenous basal release of
prostaglandins and hence is important to the physiological functions of
prostaglandins such as Gl integrity and renal blood flow. On the other
hand, it has been suggested that COX-2 is mainly responsible for the
pathological effects of prostaglandins, where induction of the enzyme
occurs in response to inflammatory agents, hormones, growth factors and
cytokines. See, U.S. Pat. 5,604,253, incorporated herein by reference, for
a discussion of the advantages of selective COX-2 inhibition. Principally,
a selective COX-2 inhibitor is expected to possess similar antiinflammatory,
antipyretic and analgesic properties to a conventional NSAID
but with reduced potential for gastrointestinal toxicity, and a reduced
potential for renal side effects.
The differential tissue distribution of COX-1 and COX-2
provides an approach to develop selective inhibitors for COX-2 with
reduced effect on COX-1, thereby preventing gastric side effects.
A number of selective COX-2 inhibitors have been reported.
These include diaryl heterocyclics (Penning etal., J. Med. Chem, 40,1347-
1365 (1997); acetoxyphenyl alkyl sulfides (Kalgutkar etal., J. Med. Chem,
41,4800-4818 (1998); methane sulfonanilides (Li etal., J. Med. Chem, 38,
4897-4905 (1995); and tricyclic inhibitor classes (Wilkerson etal., J. Med.
Chem., 38, 3895-3901 (1995). U.S. Pat. 5,604,253 discloses Nbenzylindol-
3-yl propanoic acid derivatives as cyclooxygenase inhibitors.
What is needed are additional COX-2 inhibitors, particularly
compounds which selectively inhibit the cyclooxygenase activity of COX-2
Summarv of the invention
It is an object of the invention to provide compounds and
pharmaceutical compositions thereof for inhibiting the biological activity of
COX-2, in particular the cyclooxygenase activity of COX-2.
It is an object of the invention to provide for methods of
treating disease conditions which are associated with undesired
prostaglandin production and/or secretion.
It is an object of the invention to provide for the treatment of
It is an object of the invention to provide compounds which
selectively inhibit COX-2 over COX-1.
It is an object of the invention to provide methods for
synthesizing compounds of the invention and intermediates thereof.
These and other objects of the invention shall become
apparent from the following disclosure.
Compounds of formula I, and pharmaceutically acceptable
salts thereof, are provided
X is selected from the group consisting of CrC6 trihalomethyl,
preferably trifluoromethyl; C,-C6 alkyl; and an optionally substituted or disubstituted
phenyl group of formula II:
R3 and R4 are independently selected from the group
consisting of hydrogen, halogen, preferably chlorine,
fluorine and bromine; hydroxyl; nitro; CrC6 alkyl,
preferably CrC3 alkyl; C,-C6 alkoxy, preferably CrC3
alkoxy; carboxy; C,-C6 trihaloalkyl, preferably
trihalomethyl, most preferably trifluoromethyl; and
Z is selected from the group consisting of substituted and
The carbon chains in the alkyl and alkoxy groups which may
occur in the compounds of the invention may be straight or branched. The
expression aCrC6 alkyl" thus extends to alkyl groups containing one, two,
three, four, five or six carbons. The expression "C,-C6 alkoxyl" thus
extends to alkoxy groups containing one, two, three, four, five or six
The term "aryl", alone or in combination, means a carbocyclic
aromatic system containing one, two or three rings wherein such rings may
be attached together in a pendent manner or may be fused. The term
"an/I" is intended to include not only aromatic systems containing only
carbon ring atoms but also systems containing one or more non-carbon
atoms as ring atoms. Such systems may be known as "heteroaryl11
systems. The term "aryl" is thus deemed to include "heteroaryl".
Preferred aryl groups Z include phenyl and heteroaryl, which
may be substituted or unsubstituted. By "substituted" is meant any level of
substitution, although mon- di- and tri-substitution are preferred. The
substituents are independently selected. The substituents are preferably
selected from the group consisting of halogen, particularly chlorine, fluorine
and bromine; hydroxyl; nitro; C,-C6 alkyl, preferably CrC3 alkyl, most
preferably methyl; C,-C6 alkoxy, preferably CrC3 alkoxy, most preferably
methoxy; carboxy; CrC6 trihaloalkyl, preferably trihalomethyl, most
preferably trifluoromethyl; and cyano. Although mono-, di- and trisubstitution
is preferred, full substitution, particularly when the aryl group
is phenyl, is possible. According to one preferred embodiment, Z is phenyl,
and is mono-, di-, tri-, tetra- or penta-substrtuted with halogen. The
halogen atoms may be the same or different.
According to another embodiment, Z is an aryl group other
than phenyl or substituted phenyl, and is particularly substituted or
unsubstituted heteroaryl. Such heteroaryl radicals include, for example,
pyridyl, particularly 2-, 3- and 4-pyridyl; thienyl, particularly 2- and 3-thienyi;
furyl, particularly 2- and 3-furyl; indolyl, particularly 3-, 4-, 5-, 6-, 7- and 8-
indolyl; benzothienyl, particularly 3-, 4-, 5-, 6-, 7- and 8-benzothienyl;
benzofuryl, particularly 3-, 4-, 5-, 6-, 7- and 8 benzofuryl; imidazolyl,
particularly 2- and 5-imidazolyl; pyrazolyl, particularly 3- and 5-pyrazolyl; 2-
thiazolyl; 2-benzothazolyl; quinolinyi, particularly 2-, 3-and4-quinolinyl; and
4-(2-benzyloxazolyl). Representative preferred substituted heteroaryl
groups include 6-methyl-2-pyridyl, 5-halo-2-thienyl, 5-methyl-2-thienyl, 5-
halo-2-furyl, 5-halo-3-furyl, 2,5-dimethyl-3-thienyl and 2,5-dimethyl-3-furyl.
According to one preferred embodiment of the invention, Z is an
optionally 2- or 4-substituted (or 2-, 4-di-substituted) phenyl group of the
wherein R, and R2 are independently selected from the group consisting of
hydrogen; halogen, particularly fluorine, bromine and chlorine; hydroxyl;
nitro; CrC6 alkyl; CrC6 alkoxy; and carboxy.
According to another preferred embodiment, wherein X is optionally
mono- or di-substituted phenyl according to formula II, R3 and R4 are
independently selected from the group consisting of hydrogen, halogen,
hydroxyl; nitro; CrC6 alkyl, CrC6 alkoxy and carboxy, most preferably
hydrogen, fluorine, bromine, chlorine, C,-C3 alkyl, C,-C3 alkoxy, hydroxy
and nitro. When R3 is hydrogen and R4 is other than hydrogen, the
preferred ring attachment position of R4 is the 2- or 4- position, most
preferably the 4-position. Where both R3 and R4 are other than hydrogen,
the preferred positions of substitution are the 2- and 4-positions, or the 3-
The invention is also directed to isolated optical isomers of
compounds according to formula I or V. By "isolated" means a compound
which has been substantially purified from the corresponding optical
isomer(s) of the same formula. Preferably, the isolated isomer is at least
about 80%, more preferably at least 90% pure, even more preferably at
least 98% pure, most preferably at least about 99% pure, by weight.
The invention is also directed to novel intermediates of the formula
where X and Z are defined as above.
The invention is also directed to methods for preparing the aforesaid
novel intermediates. A method for preparing a compound of formula IV
(a) reacting a ketone compound selected from the group consisting
(i) 1 , 1 , 1 -trihaloacetone, preferably 1,1,1 -trifluoroacetone; and
(ii) a compound of the formula
wherein X is CrC6 alkyl, or a radical of the formula
wherein R3 and R4 are defined above;
with a compound of the formula
wherein Z is selected from the group consisting of substituted
and unsubstituted aryl; and
(b) isolating a compound according to formula IV from the reaction
products. According to a preferred embodiment, the reaction temperature
is maintained in the range of from about 15°C to about 30°C, but higher
temperatures are possible depending on the boiling points of the reactants.
An alternative method is provided for preparing the aforesaid
intermediates of formula IV wherein X is trihalomethyl, preferably trifluoro-,
tribromo-, or trichloromethyl. The method comprises:
(a) reacting diethyl methylphosponate with an Nphenyltrihaloacetimidoyl
chloride and a compound of the formula
wherein Z is selected from the group consisting substituted and
unsubstituted aryl; and
(b) isolating a compound according fdrmuia IV wherein X is
trihalomethyl from the reaction products.
According to another embodiment of the invention, a compound of
the formula V is provided:
X is selected from the group consisting of trihalomethyl, CrC6 alkyl,
and a group of formula II:
R3 and R4 are independently selected from the group
consisting of hydrogen; halogen; hydroxyl; nitro; CrC6 alkyl;
CrC6 alkoxy; carboxy; C,-C6 trihaloalkyl; and cyano;
Z is substituted or unsubstituted aryl, preferably substituted or
unsubstituted heteroaryl; and
R5 is selected from the group consisting of
wherein Rg is (VC6 alkyl and M is Na, K or Li; or a pharmaceutically
acceptable salt thereof.
Methods are also provided for preparing compounds according to
formula I, by reacting the formula IV intermediate, wherein X and Z are
defmed as above, with 4-sulfamyl phenyl hydrazine or salt thereof; and
isolating a compound according to formula I from the reaction products.
The invention is also directed to a pharmaceutical composition of
one or more compounds of formula I in combination with a
pharmaceutically effective carrier.
According to yet another embodiment of the invention, a method for
treating a cyclooxygenase-mediated disease is provided comprising
administering an effective amount of a compound according to formula I to
an animal in need of such treatment. The expression "animal" is inclusive
of human beings.
. Description of the Figure
Fig.1 shows the inhibition of colorectal cancer cell colony growth in
the presence of compounds of the invention, as compared to celecoxib.
Detailed Description of the Invention
The compounds of formula I are potent inhibitors of COX-2. COX-2
activity was demonstrated by a cell-free assay in which human recombinant
COX-2 was incubated with test compound and [14C]-arachidonic acid. The
resulting radiolabeled prostanoid compounds, i.e., the products of COX-2
reaction with arachidonic acid, were quantified.
The compounds of the invention may be prepared via an
intermediate of formula IV:
wherein X and Z are defined as above.
The compounds of formula I are prepared by reacting the
intermediate of formula IV with sulfamyl phenyl hydrazine hydrochloride.
According to another embodiment of the invention a compound
according to formula I may be further reacted with an anhydride of the
or an acylating compound of the formula
wherein R6 is C.,-C6 alkyl, to form the corresponding sulfonamide, that is,
a compound according to formula V:
and Re is defined as above. The corresponding alkali metal salt, that is, a
compound where R5 is
and M is Na, K or Li, may be formed by reacting the above sulfonamide
with an alkali hydroxide, selected from the group consisting of NaOH, KOH
The following are general procedures for preparation of the formula
I compounds or intermediates thereof:
Procedure 1: Synthesis of trans-1,1,1-trifluoro-4-aryl-3-
To a solution of 10% sodium hydroxide in ethanol (25 ml), 1,1,1-
trifluoroacetone (10 mmol) is added and stirred at 15-20°C. To this a
solution of the appropriate araldehyde (10 mmol)
where Z is defined as above, is added and stirred vigorously for 4 hrs. The
temperature of the-reaction is maintained at 15-20°C throughout the
reaction. The solution is then poured into ice water and acidified with
concentrated hydrochloric acid. The resulting separated frans-1,1,1-
trifluoro-4-aryl-3-buten-2-one of formula IV (X=CF3) is extracted with ether
dried over anhydrous MgSO4. Evaporation of the dried ethereal layer yields
the frans-1,1,1-trifiuoro-4-aryl-3-buten-2-one which is purified by
Procedure 1 A: Alternative synthesis of frans-1,1,1-
To a cooled solution of (-70°C) lithium diisopropylamide (10 mmol), diethyl
methylphosphonate (5 mmol) is added; After the mixture is stirred for 30
minutes at -70°C, N-phenyltrifluoroacetimidoyl chloride (5 mmol) is
gradually added and stirring is continued at -70°C for 1 hour. The
appropriate araldehyde (Smmol)
where Z is defined as above, is added dropwise for 10 minutes. The
resulting mixture is warmed to room temperature over 2 hours and then
stirred overnight. Then 20 ml of dilute hydrochloric acid is added and
stirred at room temperature for 4 hours. The solution is extracted thrice
with diethyl ether (20 ml each time) and washed successively with 5%
sodium bicarbonate and brine until the pH of the solution is 6. The ethereal
layer is separated, dried over anhydrous sodium sulfate and concentrated
under reduced pressure to yield crude trans -1,1,1 -trifluoromethyl -4-aryl-3-
buten-2-one. The product is purified either by column chromatography or
The appropriate 1,1,1-trihaloacetone can be substituted for 1,1,1-
trifluoroacetone in Procedure 1 to provide othertrans-1,1,1-trihalo-4-aryl-3-
buten-2-one intermediate. Similarly, other N-phenyltrihaloacetimidoyl
chlorides can be substituted for N-phenyltrifluoroacetimidoyl chloride in
Procedure 1A to produce other trans-1,1,1-trihalo-4-aryl-3-buten-2-one
Procedure 2: Synthesis of frans-1-(alkyl or optionally
substituted aryl)-3-aryl-2-propen-1 -one
To a solution of 10% sodium hydroxide in ethanol (25 ml), a ketone
of the formula
wherein X is CrC6 alkyl (20 mmol), or a radical of formula II
wherein R3 and R4 are defined as above (10 mmol), is added and stirred at
15-20°C. To this a solution of the appropriate araldehyde (10 mmol)
where Z is defined as above, is added and stirred vigorously for 4 hours.
The temperature of the reaction is maintained at 15-20°C throughout the
reaction. The solution is then poured into ice water and acidified with
concentrated hydrochloric acid. The resulting separated frans-1-(alkyl or
optionally substituted aryl)-3-aryl-2-propen-1-one of formula IV (X - CrC6
alkyl, or radical of formula II) is extracted with ether dried over anhydrous
MgSO4. Evaporation of the dried ethereal layer yields the frans-1-(alkyl or
optionally substituted aryl)-3-aryl-2-propen-1-one, which is purified by
distillation or recrystallization.
Procedures: Synthesis of 1-(4-sulfamylaryl)-3-
To a solution of a frans-1,1l1-trifluoro-4-aryl-3-butene-2-one (5
mmol) of formula IV (X = CF3) in absolute methanol is added 4-sulfamyl
phenyl hydrazine hydrochloride (6 mmol). The mixture is refluxed with
15 stirring overnight on a hot plate with a stirrer. The solution is cooled and
poured onto crushed ice and solid material is separated by filtration.
Recrystallization of the solid material with appropriate solvent yields the
pure 1-(4-sulfamylaryl)-3-trifluorornethyl-5-aryl-2-pyrazoline of formula la:
Procedure4: Synthesis of 1-(4-sulfamylaryl)-3-
To a solution of a frans-1-(alkyl or optionally substituted aryl)-3-aryl-
2-propen-1-one (5 mmol) of formula IV in absolute methanol is added 4-
sulfamyl phenyl hydrazine hydrochloride (6 mmol). The mixture is retluxed
with stirring overnight on a hot plate with a stirrer. The solution is cooled
and poured onto crushed ice and solid material is separated by filtration.
Recrystallization of the solid material with appropriate solvent yields the
pure 1-(4-sulfamylaryl)-3-(alkyl or optionally substituted aryl)-5-aryl-2-
pyrazoline of formula I, wherein X is defined as in Procedure 2.
Procedures: Synthesis of N - [ 4 - ( 5 - A r y l - 3 -
t r i f l u r o m e t h y l p y r a z o l i n - 1 -y I)
To a solution of a 1-(4-sulfamylphenyl)-3-trifluoromethyl-5-aryl-2-
pyrazoline (10 mmol) in tetrahydrofuran (40 ml), acetic anhydride (20
mmol), 4-dimethylaminopyridine (10 mmol) and triethylamine (11 mmol) is
added and stirred for 16 hours at room temperature. The reaction mixture
is then poured into water (100 ml) and extracted with ethyl acetate. The
ethyl acetate layer is separated, washed successively with water, brine and
then dried over anhydrous sodium sulfate. The dried organic layer is
filtered and evaporated under reduced pressure to yield crude N-[4-(5-
aryl-3-trif I uromethy I pyrazol in-1-y I) phenylsulfonyljacetamide.
Recrystallization from a mixed solvent yields a pure compound.
Other sulfonamides may be prepared by substituting an anhydride
of the formula
where Rg is CrC6 alkyl, for acetic anhydride in Procedure 5 to yield
compounds of the formula VI, wherein X is trifluoromethyl:
SO2NHCR Procedures: Synthesis of N - [ 4 - ( 5 - A r y I - 3 -
t r i f l u r o m e t h y l p y r a z o l i n - 1 - y l )
phenylsulfonyljacetamide sodium salt
To a solution of N-[4-(5-aryl-3-trifluoromethylpyrazolin-1-yl)
phenylsulfonyljacetamide (5 mmol) in ethanol (100ml), sodium hydroxide
(5 mmol in 20 ml of water) is added and stirred for 5 hours. The solution
is then concentrated in vacuum to give a solid hydrated sodium salt of 1-
Salts of other sulfonamides may be prepared in the same manner
by substituting the appropriate amide according to formula V as the starting
Procedure?: Synthesis of N-[4-(5-Aryl-3-[alkyl or
optionally substituted aryl]pyrazolin-1-yl)
N-[4-(5-Aryl-3-[alkyl or optionally substituted aryl]pyrazolin-1-yl)
phenylsulfonyl]acetamides according to formula V (X = CrC6 alkyl or
optionally substituted or di-substituted phenyl) are prepared according to
Procedure 5, substituting the appropriate 1-(4-sulfamylphenyl)-3-(alkyl or
optionally substituted phenyl)-5-aryl-2-pyrazoline for 1 -(4-sutfamylphenyl)-3-
trifluoromethyl-5-aryl-2-pyrazoline as the staring material.
In similar fashion, sulfonamides according to formula V (X = CrC6
alkyl or optionally substituted or di-substituted phenyl), other than
acetamides, may be prepared by substituting the appropriate anhydride for
acetic anhydride in Procedure 5. These compounds may be converted to
salts according to Procedure 6.
The compounds of the invention preferably are characterized by a
selectivity ratio for COX-2 inhibition over COX-1 inhibition of at least about
50, more preferably at least about 100. COX inhibition may be determined
in vitro by enzyme assays well-known to those skilled in the art, such as the
enzyme assay method described later herein.
The compounds of the present invention may take the form or
pharmaceutically acceptable salts. The term "pharmaceutically acceptable
salts", embraces salts commonly used to form alkali metal salts and to form
addition salts of free acids or free bases. Where reference is made to
"compound of formula I (or formula V)" or a "compound of the invention",
it is understood that pharmaceutically acceptable salts are also included.
The nature of the salt is not critical, provided that it is pharmaceuticallyacceptable.
Suitable pharmaceutically acceptable acid addition salts may
be prepared from an inorganic acid or from an organic acid. Examples of
such inorganic acids are hydrochloric, hydrobromic, hydroiodic, nitric,
carbonic, sulfuric and phosphoric acid. Appropriate organic acids may be
selected from aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic,
carboxylic and sulfonic classes of organic acids, example of which are
formic, acetic, propionic, succinic, glycolic, gluconic, lactic, malic, tartaric,
citric, ascorbic, glucuronic, maleic, fumaric, pyruvic, aspartic, glutamic,
benzoic, anthranilic, mesylic, salicyclic, salicyclic, 4-hydroxybenzoic,
phenylacetic, mandelic, embonic (pamoic), methanesulfonic,
ethanesulfonic, benzenesulfonic, pantothenic, 2-hydroxyethanesulfonic,
toluenesulfonic, sulfanilic, cyclohexylaminosulfonic, stearic, algenic, betahydroxybutyric,
salicyclic, galactaric and galacturonic acid. Suitable
pharmaceutically acceptable base addition salts of compounds of formula
I include metallic salts made from calcium, lithium, magnesium, potassium,
sodium and zinc or organic salts made from N.N'-dibenzylethylenediamine,
chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (Nmethylglucamine)
and procaine. All of these salts may be prepared by
conventional means from the corresponding compound of formula I or V by
reacting, for example, the appropriate acid or base with the compound of
formula I or V.
The compounds of the present invention may be administered in the
form of a pharmaceutical composition, in combination with a
pharmaceutically acceptable carrier. The active ingredient in such
formulations may .comprise from 0.1 to 99.99 weight percent. By
"pharmaceutically acceptable carrier" is meant any carrier, diluent or
excipient which is compatible with the other ingredients of the formulation
and to deleterious to the recipient.
The compounds of the invention may be administered to individuals
(animals, most particularly mammals including humans) afflicted with any
disorder characterized by undesirable prostaglandin production resulting
from cyclooxygenase activity, particularly COX-2 activity ("cyclooxygenasemediated
disorder"). In particular, the compounds of the invention are
believed useful in treating inflamation and inflamation-related disorders, by
administering to a subject having or susceptible to such inflamation or
inflamation-related disorder and effective amount of a compound according
to formula I. Inflamation is associated with a variety of disease conditions.
For a list of such disease conditions treatable by cyclooxygenase inhibitors,
and COX-2 inhibitors in particular, see U.S. Patents 5,604,253 and
5,908,852, the entire disclosures of which are incorporated herein by
reference. Such conditions include, for example, arthritis, including but not
limited to rheumatoid arthritis, spondyloarthropathies, gouty arthritis,
osteoarthritis, systemic lupus erythematosus and juvenile arthritis. Such
conditions further include rheumatic fever, symptoms associated with
influenza or other viral infections, common cold, low back and neck pain,
dysmenorrhea, headache, toothache, sprains and strains, myositis,
neuralgia, synovitis, gout and ankylosing spondylitis, bursitis, and following
surgical and dental procedures. The compounds of the invention are
believed useful as analgesics for treating or alleviating all forms of pain.
The compounds are believed useful in the treatment of other disorders
including asthma, bronchitis, tendinitis, bursitis; skin related conditions such
as psoriasis, eczema, bums and dermatitis; gastrointestinal conditions such
as inflammatory bowel disease, Crohn's disease, gastritis, irritable bowel
syndrome and ulcerative colitis and for the prevention of colorectal cancer;
the treatment of inflamation in such diseases as vascular diseases,
migraine headaches, periarteritis nodosa, thyroiditis, aplastic anemia,
Hodgkin's disease, sclerodoma, type I diabetes, myasthenia gravis,
sarcoidosis, nephrotic syndrome, Behcet's syndrome, polymyositis,
gingivitis, hypersens'rtivity, conjunctivitis, swelling occurring after injury,
myocardial ischemia, and the like. The compounds of the invention are
believed useful as antipyretics for the treatment of fever.
In addition, compounds of the invention may inhibit cellular
neoplastic transformations and metastatic tumor growth and hence can be
used in the treatment of cancer. In particular, the present invention
provides a method for treating or preventing a neoplasia that produces a
prostaglandin in a subject in need of such treatment or prevention, the
method comprises treating the subject with a therapeutically effective
amount of a compound of formula I or V. The term "neoplasia" includes
neoplasia that produce prostaglandins or express a cyclooxygenase,
including both benign and cancerous tumors, growths and polyps.
Neoplasias believed treatable with cyclooxygenase inhibitors are discussed
in U. S. Pat. 5,972,986, the entire disclosure of which is incorporated
herein by reference. The compounds may be used to inhibit the growth or
an established neoplasm, i.e., to induce regression, or to prevent or delay
the onset of the neoplasm.
According to U.S. Pat. 5,972,986, neoplasias that produce
prostaglandins, and which are therefore believed treatable with the
compounds of the invention, include brain cancer, bone cancer, epithelial
cell-derived neoplasia (epithelial carcinoma) such as basal cell carcinoma,
adenocarcinoma, gastrointestinal cancer such as lip cancer, mouth cancer,
esophageal cancer, small bowel cancer and stomach cancer, colon cancer,
liver cancer, bladder cancer, pancreas cancer, ovary cancer, cervical
cancer, lung cancer, breast cancer and skin cancer, such as squamous cell
and basal cell cancers, prostate cancer, renal cell carcinoma, and other
known cancers that effect epithelial cells throughout the body.
The compounds of the invention may also be useful in the treatment
of angiogenesis-mediated disorders. Thus, a method for treating, inhibiting
or delaying the onset of an angiogenesis-mediated disorder in a subject is
provided comprising administering to a subject in need of such treatment
an effective amount of a compound according to the present invention.
Angiogenesis-mediated disorders which may be treatable with
cyclooxygenase inhibitors are discussed in U. S. Pat. 6,025,353, the entire
disclosure of which is incorporated herein by reference. According to U. S.
Pat. 6,025,353, such disorders include, for example, metastasis, comeal
graft rejection, ocular neovascularization, retinal neovascularization,
diabetic retinopathy, retrolental fibroplasia, neovascular glaucoma, gastric
ulcer, infantile hemaginomas, angiofibroma of the nasopharynx, avascular
necrosis of bone, and endometriosis.
The compounds may be administered by any route, including oral
and parenteral administration. Parenteral administration includes, for
example, intravenous, intramuscular, intraarterial, intraperitoneal,
intranasal, rectal, or subcutaneous administration. The active agent is
preferably administered with a pharmaceuticaliy acceptable carrier selected
on the basis of the selected route of administration and standard
The active agent may be formulated into dosage forms according to
standard practices in the field of pharmaceutical preparations. See
Alphonso Gennaro, ed., Remington's Pharmaceutical Sciences, 18th Ed.,
(1990) Mack Publishing Co., Easton, PA. Suitable dosage forms may
comprise, for example, tablets, capsules, solutions, parenteral solutions,
troches, suppositories, or suspensions.
For parenteral administration, the active agent may be mixed with
a suitable carrier or diluent such as water, an oil, saline solution, aqueous
dextrose (glucose) and related sugar solutions, or a glycol such as
propylene glycol or polyethylene glycol. Solutions for parenteral
administration preferably contain a water soluble salt of the active agent.
Stabilizing agents, antioxidizing agents and preservatives may also be
added. Suitable antioxidizing agents include sulfite, ascorbic acid, citric
acid and its salts, and sodium EDTA. Suitable preservatives include
benzalkonium chloride, methyl- or propyl-paraben, and chlorbutanol.
For oral administration, the active agent may be combined with one
or more solid inactive ingredients for the preparation of tablets, capsules,
or other suitable oral dosage forms. For example, the active agent may be
combined with carboxymethylcellulose calcium, magnesium stearate,
mannitol and starch, and then formed into tablets by conventional tableting
The specific dose of compound according to the invention to obtain
therapeutic benefit will, of course, be determined by the particular
circumstances of the individual patient including, the size, weight, age and
sex of the patient, the nature and stage of the disease, the aggressiveness
of the disease, and the route of administration. For example, a daily
dosage of from about 0.01 to about 150 mg/kg/day may be utilized. Higher
or lower doses are also contemplated.
The compounds of the present invention are optically active due to
the presence of a chiral carbon atom at position 5 of the pyrazoline
Other chiral carbon atoms may also be present. The present invention is
meant to comprehend diastereomers as well as their racemic and resolved,
enantiomerically pure forms and pharmaceutically acceptable salts thereof.
Isolated optical isomers may be purified from racemic mixtures by wellknown
chiral separation techniques. According to one such method, a
racemic mixture of a compound having the structure of formula I or V, or
chiral intermediate thereof, is separated into 99% wt.% pure optical isomers
by HPLC using a suitable chiral column, such as DA1CEL CHIRALPAK AD
(Daicel Chemical Industries, Ltd., Tokyo, Japan). This column contains a
packing of amylosetris(3,5-dimethylphenyl) carbamate coated on a 10 pm
silica-gel substrate. The column has a size of 250 x 4.6 mm (L x I.D.). The
column is operated according to the manufacturer's instructions. A flow
rate should be maintained that will result in column pressures of less than
430 psi (30 kg/cm2). A typical flow rate is 1.0 ml/min. The operating
temperature range is 0°C - 40°C. The maximum operating pressure is
1200 psi. One suitable mobile phase system is hexane/2-propanol (100/0
to 0/100 v/v). A typical hexane/2-propanol mobile phase is hexane/2-
propanol (90/10 v/v). Another suitable mobile phase system is
hexane/ethanol (100/0 to 85/15 v/v), (40/60 to 0/100 v/v). Suitable mobile
phase modifiers include N.N-diethylamine for a basic sample, and
trifluoroacetic acid for an acidic sample.
The practice of the invention is illustrated by the following nonlimiting
1 - (4-sulfamylphenyl)-3-trifluoromethyl-5-phenyl-2-pyrazoline
A. Trans-1,1,1-trifluoro-4-phenyl-3-buten-2-one was prepared
according to Procedure 1 from 1,1,1-trifluoroacetone and benzylaldehyde.
B. A solution of frans-1,1,1-trifluoro-4-phenyl-3-buten-2-one (5
mmol) and 4-sulfamylphenyl hydrazine hydrochloride (6 mmol) was
subjected to Procedure 3. The title compound was obtained in 73% yield,
m.p. 132-135°C; C, H analysis (C18H1SS02N
Table 1, Examples 2-23, lists additional compounds which are
prepared by reacting a frans-1,1,1 -trifluoro-4-(substituted)phenyl-3-buten-2-
one (5 mmol) and 4-sulfamylphenyl hydrazine hydrochloride according to
A. Trans-l.l.l-trifluoro-^S-indolyO-S-buten^-one was prepared
according to Procedure 1 from 1,1,1-trifluoroacetone and 3-indolyl
B. A solution of frans-1,1,1 -trifluoro-4-(3-indolyl)-3-buten-2-one (5
mmol) and 4-sulfamylphenyl hydrazine hydrochloride (6 mmol) was
subjected to Procedure 3. The title compound was obtained in 82% yield,
m.p. 178-180 °C; C, H analysis (C16H14S02N4F3):
Table 2, Examples 25-30, lists additional compounds which are
prepared by reacting £rans-1,1,1-trifluoro-4-aryl-3-buten-2-one and 4-
sulfamylphenyl hydrazine hydrochloride according to Procedure 3.
Table 3, Examples 31-40, lists additional compounds which were
prepared according to Procedures 2 and 4.
Cyclooxygenase Enzyme Assay
Compounds were tested for inhibitory activity against COX-1 and
COX-2. The compounds of Examples 1 and 24 had the highest selectivity
for inhibiting COX-2.
Cyclooxygenase activity of ovine COX-1 (Oxford Biomedical
Research Inc.) and human recombinant COX-2 (Oxford Biomedical
Research Inc.) was assayed by a thin layer chromatography (TLC) method
as follows. All inhibitors were dissolved in dimethyl sulfoxide to a stock
solution of 5mM. Human recombinant COX-2 (3 units) or ovine COX-1 (15
units) was incubated with inhibitors at several concentrations in a solution
containing 100 mM Tris-HCI, pH7.8, 500 uM phenol and hematin for 90
to 120 minutes at room temperature (24°C). In controls, equal volumes of
DMSO without drug were added to the incubation mixture. After incubation
for 90-120 minutes, [1-14C] arachidonic acid (50uM, 51mCi/mmo!) (DuPont
MEN) was added and incubated at 37°C for 2 minutes. The reaction was
terminated by extraction with 1 ml of ethyl acetate. The ethyl acetate layer
was transferred into a fresh tube and evaporated to dryness in a Speedvac
vacuum dryer. The contents of the tubes were reconstituted in 20 ml of
ethyl acetate and spotted on a TLC plate (J.T. Baker, Phillipsburg, NJ) and
developed in a mobile phase containing chloroform/methanol (95:5) at4°C.
Radiolabeled prostanoid compounds (the products of COX enzymatic
reaction with radiolabeled arachidonic acid substrate) were quantitated with
a radioactivity scanner (Fuji, Phosphorimager). The percentage of total
products observed at different inhibitor concentrations was divided by the
percentage of the products observed for protein samples pre incubated for
the same time with DMSO. The results are shown in Table 4. The
Example 1 and 2 compounds are more than one thousand times more
active in inhibiting COX-2 compared to COX-1.
Table 4: Inhibition of Cyclooxygenase Activity
Ex. 2 COX-2 COX-1
1 C6H5 0.10 100
24 3-indofyl 0.078100
Soft Agar Assay
The Example 1 and 24 compounds were compared to the COX-2
inhibitor celecoxib in inhibiting the growth of DLD-1 cells in soft agar. DLD-
1 cells are human colorectal carcinoma cells that overexpress COX-2.
DLD-1 cells grow in soft agar and form tumors in nude mice. The soft agar
assay was performed as follows. A layer of bottom agar (8% noble agar)
was placed onto 60 mm2 tissue culture dishes. The tumor cells were
trypsinized from normal growth flasks while in exponential growth. The
cells were counted by using a hemacytometer and 1.0 x 105 cells were
placed into the top agar mixture containing growth medium, 4% noble agar
and various concentrations of drugs. The concentration range was
normally between 10 uM to 75 uM. The cells were not refed during the
assay system; therefore, the cells were treated with one dose of the
agents. The plates were stained 20 days later with a 0.05% (w/v) nitroblue
tetrazolium solution (which stains only viable cells) for 48 hours. The
results are shown in Fig. 1, the y-axis being the percent of cell colonies
remaining in comparison to untreated control cells. Even at the highest
concentration tested, celecoxib obtained only about partial inhibition,
compared to 100% for the compounds of the invention.
All references cited herein are incorporated herein by reference.
The present invention may be embodied in other specific forms
without departing from the spirit or essential attributes thereof and,
accordingly, reference should be made to the appended claims, ratherthan
to the foregoing specification, as indication the scope of the invention.
1. A l-(4-sulfamylaryl)-3-trifluoromethyl-5-aryl-2-pyrazoline compound of the formula:
Z is a substituted or unsubstituted aryl other than substituted or unsubstituted phenyl; and
wherein said aryl includes carbocyclic aromatic systems and heteroaryl systems, which systems contain one, two or three rings wherein such rings may be attached together in a pendent manner or may be fused,
wherein said heteroaryl is selected from indolyl, furyl, thienyl, pyridyl, benzofuryl, benzothienyi, imidazolyl, pyrazolyl, thiazolyl, benzothiazolyl, quinolinyl, and 4-(2-benzyloxazolyl);
or a pharmaceutically acceptable salt of such a compound.
2. A compound as claimed in claim 1 wherein Z is 3-indolyl; or a pharmaceutically
acceptable salt thereof.
3. The compound as claimed in claim 2 which is l-(4-sulfamylphenyl)-3-
trifluoromethyl-5-(3-indolyl)-2-pyrazoline; or a pharmaceutically acceptable salt thereof.
4. A compound as claimed in any of claims 1 to 3, as and when used for preparation
of a pharmaceutical composition.
|Indian Patent Application Number||IN/PCT/2002/0001/DEL|
|PG Journal Number||40/2008|
|Date of Filing||01-Jan-2002|
|Name of Patentee||TEMPLE UNIVERSITY- OF THE COMMONWEALTH SYSTEM OF HIGHER EDUCATION|
|Applicant Address||BROAD STREET AND MONTGOMERY AVENUE, PHILADELPHIA, PA 19122, U.S.A.|
|PCT International Classification Number||A61K 31/415|
|PCT International Application Number||PCT/US00/16656|
|PCT International Filing date||2000-06-16|