WO 2004/052893 PCT/US2003/038932
SUBSTITUTED DlHYDROPYRANO INDOLE-3,4-DlONE DERIVATIVES AS
INHIBITORS OF PLASMINOGEN ACTIVATOR INHIBITOR-1 (PAI-1)
This invention relates to substituted dihydnopyrano indole-3,4-dione
5 derivatives useful as inhibitors of plasminogen activator inhibitor-1 (PAI-1) and
from fibrinolytic disorders such as deep vein thrombosis and coronary heart disease,
and pulmonary fibrous.
10 BACKGROUND OF INVENTION
Plasminogen, activator inhibitor (PAI-1) is a major regulatory component of
the plasminogen-plasmin system. PAI-1 is the principal physiologic inhibitor of both
tissue type plasminogen activator (tPA) and urokinase type plasminogen activator
1 5 (uPA). Elevated plasma levels of PAI-1 have been associated with thrombotic events
as indicated by animal experiments (Krishnamurti, Blood, 69, 796 (1937); Reilly,
Arteriosclerosis and Thrombosis 11. 1276 (1991); Carrnellet, Journal of Clinical
Investigation, 92, 2756 (1993)) and clinical studies (Rocha, Fibrinolysis, 8,294, 1994;
Aznar, Haemostasis 24, 243 (1994)). Antibody neutralization of PAI-1 activity
20 resulted in promotion of endogenous thrombolysis and reperfusion (Biemond,
Circulation, 91, 1176 (1995) Levl, Circulation 85, 305, (1992)). Elevated levels of
PAI-1 have also been implicated in diseases of women such as polycystic ovary
syndrome (Nordt, Journal of clinical Endocrinology and Metabolism, 85, 4, 1563
(2000)) and bone loss induced by estrogen deficiency (Daci, Jouroal of Bone and
25 Mineral Research, 16. 8, 1510 (2000)), Accordingly, agents that inhibit PAI-1 would
be of utility in treating conditions originating from fibrinolytic disorder such as deep
vein thrombosis, coronary heart disease, pulmonary fibrosis, Alzheimer's disease,
polycystic ovary syndrome, etc.
30 WO 99/43654 and WO 99/43651 disclose indole derivatives, of the following
formuIa as inhibitors of phospholipase enzymes useful in preventing inflammatory
conditions.

WO 2004/052893 PCT/US2003/038932

5 wherein; A is a five-membered, or six-membered ring helerocycie; X is a bond, an
alkylene. or a vinylene radical; R1 is a H, alkyl, alkenyl, cycloalkyl, cycloalkenyl.
carboxyl, cyano, alkylcarbonyl, alkaxycarbonyl. amincarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl or aryl radical: R2 is H, alkyl, trihalogenomethyl, hydroxyl
cycloalkyl, cyano, carboxyl, cycloalkenyl, carboxyl, cyano, alkylcarbonyl, alkoxy-
10 carbonyl aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl or aryl radical;
and R3 is a hydrogen atom.

15 wherein R, is a H, halogen, nitro, carboxy, protected carboxy, lower alkenyl, or acyl;
R2 is H, halogen, carboxy, lower alkenyl, or acyl; R3 is a lower alkenyl, or lower
alkenyl, both optionally substituted; and R4 is carboxy, protected carboxy, or acyl.
WO 9938297 relates to substituted indoles of the following formula with
20 thrombin inhibiting effect and fibrinogen receptor antagonist effiect.
-2-

WO 2004/052893 PCT/US2003/038932

where: Ra is halogen, carboxy, R3R4N-CO-, R3R4SO2-, or R4R5N-; Rb and Rd are
either alkyl or R2-A where R2 is a phenyl optionally substituted and A is an alkylene of
a substituted alklene; and Rc is a hydrogen, or alkyl.
5
EP 0 655 439 teaches 5,6 fused ring bicyclic compounds inclusive of indoles,
benzofurans, and benzothiophenes corresponding the following formula as platelet

10 SUMMARY OF THE INVESTIGATION

15
wherein:
X is hydrogen, an alkali metal or a basic amine moitely;
R1 is, C1-C2 alkyl, preferably C1-C6 alkyl, C2-C6 cycloalkyl, -CH2-C5-C6
20 cycloalkyl, pyridinyl. -CH2-pyridinyl, phenyl or benzyl, wherein the rings of the
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WO 2004/052893 PCT/US2003/038932
Cycloalkyl, pyridinyl, phenyl and benzyl groups may be optionally substituted by from
1 to 3 groups independenty selected from, halogen, C1-C5 alkyl. C1-C6 perfluoroalkyl,
-O-C1-C6 perfluoroalkyl, C1-C6 alkoxy, -OH, -NH2. or -NO2;
5 R2 is hydrogen, halogen, C1-C6 alkyl, C1-C3 perfluoroalkyl, C1-C6 alkoxy. C3-C8
cycloalkyl, -CH2-C4-C5 cycloalkyl, hydroxy, -NH2 or-NO2;
R3 is hydrogen, halogen, C1-C6 alkyl. C1-C3 perfluoroalkyl, C1-C6, C3-C6
cycloalkyl, CH2-C3-C5 cycloalkyl, hydroxy, -NH2 -NO2, phenyl, benzyl, benzyloxy,
10 pyridinyl, or -CH2-pyridinyl, wherein the rings of these groups may be optionally
substituted by from 1 to 3 groups independently selected from phenyl, halogen, C1-C6
alkyl, C1- C6 perfluoroalkyl, -O-C1-C6 perfluoroalkyl, C1-C6 alkoxy, -OH, -NH2, or -NO2;
or a pharmaceutically acceptable salt or ester form thereof.
15 DETAILED DESCRlPTlON OF THE INVENTION

20
wherein R1, R2, and R3 are as defined above, or a pharmaceutically acceptable salt
of ester form thereof.

WO 2004/052893 PCT/US2003/038932

wherein:
5 R1 is C1-C5 alkyl, preferably C1-C6 alkyl, C3-C6 cycloalkyl, -CH2-C3-C6
cycloalkyl, or benzyl, wherein the rings of the cycloalkyl and benzyl groups may be
optionally substituted by from 1 to 3 groups independently selected from halogen, C1-
C5 alkyl. C1-C6 perfluoroalkyl. -O-C1-C6 perfluoralkyl, C1-C6 alkoxy, -OH. -NH2, or
-NO2,
10
R2 is hydrogen, halogen, C1-C6 alkyl. C1-C3 perfluoroalkyl, preferably -CF3
C1-C6 alkoxy, C3-C8 cycloalkyl, -CH2-C3-C8 cycloalkyl, hydroxy, -NH2, or-NO2
R4. R5 and R6 are each independently hydrogen, phenyl, halogen, C1-C3 alkyl,
15 C1-C3 perfluoroalkyl, preferably -CF3, -O-C1-C2 perfluoroalkyl, C1-C3 alkoxy, -OH,
-NH2, or-NO2: or a pharmaceutically acceptable salt or ester form thereof.
The alkali metals suitable for use in the present invention include: sodium,
potassium, lithium, calcium, magnesium, etc.. The basic amine moleties include:
20 amonia, primary amines, secondary amines, tertiary amines, pyridine, aromatic
amines, benzyl amines, etc. The term "alkyI" includes both straight and branched
carbon chains. The preferred C1-C3 perfluoroalkyl substituent is -CF3 and the
preferred O-C1-C3 perfluoroalkyl substituent is -0CF2.
25 The present invention further comprises a method of inhibiting in a mammal
plasminogen activator type 1 (PAI-1) comprising administering to a mammal in need
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WO 2004/052893 PCT/US2003/038932

5
X is an alkali metal or a basic amine moiety as defined above;
R1 is C1-C5 alkyl, preferably C1-C6 alkyl, C3-C6 cycloalkyl, -CH2-C3-C6 cyclo-
alkyl pyridinyl, -CH2-pyridinyl, phenyl or benzyl, wherein the rings of the cycloalkyl,
10 pyridinyl. phenyl and benzyl groups may be optionally substituted by from 1 to 3
groups selected from, halogen, C1-C3 alkyl, C1-C3 perfluoroalkyl, -O-C1-C3
perfluoroalkyl, C1-C3 aIkoxy, -OH, -NH2, or -NO2
R2 is hydrogen, halogen, C1-Ca alkyl, C1-C3 perfluonoalkyl. C1-C5 alkoxy, C3-C6
15 cycloalkyl, -CH2-C3-C5 cycloalkyl, hydroxy, -NH2, or -NO2;
R3 is hydrogen, halogen, C1-C5 alkyl. C1-C3 pperfluoroalkyl, C1-C8, C3-C6
cycloalkyl I, CH2-C3-C6 cycloalkyl. hydroxy, -NH2, -NO2, phenyl, benzyl, benzyloxy,
pyridinyl. or -CH2-pyridinyl, wherein the rings of these groups may be optionally
20 substituted by from 1 to 3 groups selected from halogen, C1-C3 alkyl. C1-C3
perfluoroalkyl, -O-C1-C3 perfluoroalkyl, C1-C3 alkoxy, -OH, -NH2. or -NO2; or a
pharmaceutically acceptable salt or ester form thereof.
The preferred salt forms of the compounds herein include but are not
25 limited to sodium salts, and potassium sails. Other useful salt forms of these
compounds include those formed with pharmaceutically acceptable inorganic and
organic bases known in the art. Salt forms prepared using inorganic bases include
hydroxides, carbonates or bicarborsates of the therapeuticalty acceptable alkail
metals or alkaline earth metals, such as sodium potassium, magnesium, calcium and
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WO 2004/052893 PCT/US2003/038932
the like. Acceptable organic bases include amines, such as benzylzmine, mono, di-
and trialkylamines preferably those having alkyl groups of from 1 to 6 carbon atoms,
more preferably 1 to 3 carbon atoms, such as methylamine, dimethylamine,
trimethylamine, ethylamine, diethylamine, triethylamine, mono-, di-, and tirethanol-
5 amine. Also useful are alkylene diamines containing up to 6 carbon atoms, such as
hexamethylenediamine; cyclic saturated or unsaturated bases containing up to 6
carbon atoms, including pyrrolidine, peperidine, morpholine, piperazine and their N-
alkyl and N-hydroxyalkyl derivatives, such as N-methyl-morpholine and N-(2-
hyroxyethyl)-piperidine, or pyridine. Quaternary salts may also be formed, such as
10 tetralkyl forms, such as tetramethyl forms, alkyl-alkanol forms, such as methyl-
triethanol or trimethyl-monoethanol forms, and cyclic ammonium salt forms, such as
morpholinium, N-methyl-N-(2-hydroxyethyl)-morpholinium, or N,N-dimethyI-
piparidinium salt forms. These salt forms may be prepared using the acidic
15 compound(s) of Formula I and procedures known in the art.
Ester forms of the compounds of this invention include straight chain alkyI
esters having from 1 to 6 carbon atoms or branched chain alkyl groups containing 3
or 6 carbon atoms. Including methyl, ethyl, propl, butyl, 2-methylpropyl and 1,1-
20 dimethylethyl esters. Other esters useful with this invention include those of the
formula -COOR7- wherein R7 is selected from the formulae:

wherein R8, R9, R10, R11 are independently selected from hydrogen, alkyl of from 1 to
10 carbon atoms, aryl of 6 to 12 carbon atoms. arylalikyl of from 6 to 12 carbon
25 atoms; hetergoryl or alkyheteroaryl wherein the heteroaryl ring is bound by an alkyI
chain of from 1 to 6 carbon atoms.
Among the preferred ester forms of the compounds herein include but not
limited to C1-C8 alkyl esters, C3-C6 branched alkyl esters, benzyl esters, etc.
30
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wo 2004/052893 PCT/US2003/038932
As used herein. "aryl" refers to an unsaturated aromatic carbocyclic group of
from 6 to 14 carbon atoms having a single ring (e.g., phenyl) or multiple condensed
(fused) rings (e.g., naphthyl or anthryl). Preferred aryl groups include phenyl,
naphthyl and the like. As used herein, "heteroaryl" refers to an aromatic heterocyclic
5 ring system (monocyclic or bicyclic) where the heteroaryl moieties are five or six
membered rings containing 1 to 4 heteroatoms selected from the group consisting of
S, N, and O, Such heteroaryl groups can have a single ring, such as pyridyt, pyrrolyl
or furyl groups, or multiple condensed rings, such as indolyl, indolizinyl, benzofuranyl
or benzothienyl groups. Preferred heteroaryls include pyridyl, pyrrotyl and furyl.
10
Unless otherwise limited by the definition for the aryl or heteroaryl groups
herein, such groups can optionally be substituted with from 1 to 5 substituents
selected from the group consistinjg of acyloxy, hydroxy, acyl, alkyl of 1 to 6 carbon
atoms, alkoxy of 1 to 6 carbon atoms, alkenyl of 2 to 6 carbon atoms. alkynyl of 2 to 6
15 carbon atoms, substituted alkyl, substituted alkoxy, substituted alkenyl, substituted
alkynyl, amino, amino substituted by one or two alkyl groups of from 1 to 6 carbon
atoms, aminoacyl, acylamino, azido, cyano, halo, nitro, thioalkoxy of from 1 to 6
carbon atoms, substituted thloalkoxy of from 1 to 6 carbon atoms, amd trihalomethyl.
Substituents on the alkyl, alkenyl, alkynyl, thioalkoxy and alkoxy groups mentioned
20 above include halogens, CN, OH, and amino groups. Preferred substituents on the
aryl groups herein include alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon
atoms, halo, cyano, nitro, trihalomethyl, and thioalkoxy.
The compounds of the present invention are inhibitors of the serine protease
25 inhibitor PAI-1, and are therefore useful in the treatment, inhibition, prevention or
prophylaxis in a mammal, preferably in a human, of those processes which involve
the production and/or action of PAI-1. Thus, the compounds of the invention are
useful in the treatment or prevention of noninsulin dependent diabetes melitus and
prevention of thrombotic events associated with coronary artery and cerebrovascular
30 disease. These compounds would also be useful for inhibiting the disease process
involving the thrombotic and prothrombotic states which include, but are not limited
to, formation of atherosclerotic plaques, venous and arterial thrombosis, myocardial
ischemia, atrial fibrillation, deep vein thrombosis, coagulation syndromes, pulmonary
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WO 2004/052893 PCT/US2003/03932
fibrosis. cerebral thrombosis, thromboembolic complications of surgery (such as joint
replacement), and peripheral arterial occlusion. These compounds are also useful in
treating stroke associated with aor resulting from atrial fibrillation.
5 The corripounds of the invention may also be useful in the treatment of
diseases associated with extracellular matrix accumulation. includig, but not limited
to. renal fibrosis, chronic obstructive pulmonary disease, polycystie ovary syndrome,
restenosis, renovascular disease and organ transplant rejection.
10 The compounds of the invention may also be useful in the treatment of
malignancies, and diseases associated with neoangiogenesis (such as diabetic
retinopathy).
The compounds in the invention may also be used in conjunction with and
15 following processes or procedures involving maintaining blood vessel patency,
including vascular surgery, vascular graft and stent patency, organ, tissue and cell
implantation and transplantation.
The compounds of the invenaon may also be used in the treatment of
20 Alzheimer's disease. This method may also be characterised as the inhibition of
plasminogen activator by PAl-1 in mamal, particularly a human experiencing or
subject to Alzheimer's disease. This method may also bo characterized as a method
of increasing or normalizing levels of plasmin concentration in a mammal, particularly
those experencing or subject to Alzheimer's disease.
25
The compounds of the present invention are useful for the treatment of blood
and blood products used in dialysis, blood storage in the fluid phase, especially ex
vivo platelet aggregation. The present compounds may also be added to human
plasma during the analysis of blood chemistry in hospital settings to determine the
30 fibrinolytic capacity thereof.
The compounds in the present invention may also be used in combination
with prothrombolytic, fibrinolytic and anticoagulant agent.
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WO 2004/052893 PCT/US2003/038932
The compounds of the present invention may also be used to treat cancer
including, but not limited to, breast and ovarian cancer, and as imaging agents for the
identification of metastatic cancers.
5
The compounds of the invention may also be useful in the treatment or
myelofibrosis with myelod metaplasia by regulating stromal cell hyperplasia and
increases in extracellular matrix proteins.
10 The present compounds may also be used in conjunction with protease
inhibitors containing highly active antiretroviral therapy (HAART) for the treatment of
diseases which originate from fibrinolytic impairment and by hypercoagulability of
HIV-1 infected patients receiving such therapy.
15 The compounds of the invention may be used for the treatment of diabetic
deuropathy and renal dialysis associated with nephropathy.
The compounds of the invention may be used to treat cancer, septicemia,
obesity, insulin resistance, proliferative diseases, such as psoriasis, improving
20 coagulation homeostasis, censbrovascular diseases, microvascular disease
hypertension, dementia, osteoporosis, arthritis, asthma, heart failure, arrhythmia,
angina, as a hormone replacement agent, treating, preventing or reversing
progression of atheroscierosis. Alzheimer's disease, osteopenia, reducing
inflammatory markers, reducing C-reactive protein. preventing or treating low grade
25 vascular inflammation, stroke, coronary heart disease, primary and secondary
prevention of myocardfal infarction, stable and unstable angina, primary prevention of
cardiovascular events, secondary prevention of cardiovascular events, peripheral
vascular disease, peripheral arterial disease, acute vascular syndromes reducing the
risk of undergoing a myocardial revascularization procedure, micromuscular diseases
30 such as nephropathy, neuropathy, retinapalhy, nephrolic syndrome, Type 1 and Type
2 diabetes and rented diseases, hyperglycemia, hyperinsulinemia, malignant lesions,
pre malignant lesions, gastro intestinal malignancies, liposancomas and epithelial
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WO 2004/052893 PCT/US2003/038932
lumors, proliferative diseases such as psoriasis, improving coagulation, homeostasis
and/or improving endothellal function and all forms of cerebrovascular diseases.
The compounds of the invention may also be used for topical application in
5 wound healing for the prevention of scarring.
Methods for the treatment, inhibition, prevention or prophylaxis in a
mammal of each of the conditions or maladies listed herein are part of this invention.
Each method comprises administering to a mammal in need thereof a
10 pharmaceuticaly or therapeutically effective amount of a compound of this invention,
or a pharmaceutically acceptable salt or ester fom thereof.
The compounds of the present invention can be prepared according to the
method described in Scheme 1 or modification thereof using readily available starting
15 materials, reagents and conventional synthetic procedures. It is also possible to
make use of variants of these process steps, which in themselves are known to and
well within the preparatory skill of the medicinal chemist. In the following reaction
schemes, R1, R2, R4, R5 and R6 are selected from the groups defined above.
20 In Scheme 1, the bromo-indole-2-carboxylates (II) were reacted with alkyl
halides or aryl-alkyl halides using a bass such as sodium hydride in DMF or THF to
give the N-substituted brom-indole carboxylates (III). The N-substituted bromo-
indole carboxylates (III) were then subjected to palladium catalyzed cross-coupling
with various substituted aryl-halides affording the N-substituted Aryl-indole
25 carboxylates (V). Alternatively. reaction of bromo-indole-2-Carboxylates (II) with
various substituted aryl-bromic acids under the palladium catalyzed cross-coupling
conditions afforded the aryl-indole carboxylates (IV). Alkylation of (IV) with alkyl-
halides or aryl-alkyl-halides under basic conditions as described above afforded the
N-substituted Aryl-Indole carboxylates (V). Reduction of V to the corresponding
30 alcohol (VI) was accomplished by treating V with lithium aluminum hydride in ether or
THF. Reaction, of the alcohol (VI) with acatyl chloride in presence of a base such as
triethyl amine or N,N-diisopropyl ethyl amine in an inert solvent such as methylene
chloride afforded the acetate VII. Reaction of VII with oxalyl chloride in THF or
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WO 2004/052893 PCT/US2003/038932
dichloromethane and subsequent quenching with water furnished the keto acid VIII.
Treatment of the acetate (VlIl) with an aqueous base such as KOH or NaOH in a
solvent such as THF fumished the carboxylate salt (IX). Lactomization of (IX) to the
corresponding pyrano indole derivatives (l) was accomplished by treating IX with an
5 aqueous acid such as hydrogen chloride in an organic solvent such as

10
This invention also provides pharmaceutical compositions comprising a
pharmaceutically or therapeutically effective amount of a compound of this invention,
or a pharmaceutically acceptable salt or ester form thereof, eiter alone of in
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WO 2004/03289 PCT/US2003/038932
combination with one or more pharmaceutically acceptable carriers or excipients (i.e.
pharmaceutically acceptable materials with no pharmacological effects). It will be
understood that a pharmaceutically or therapeutically effective amount of a
compound herein refers to an amount of the compound to question which will
5 sufficiently inhibit the serine protease inhibitor PAI-1 in the mammal in need thereof to
a sufficient extent to provide a desirable improvement in the condition in question or
provide sufficient inhibition of the serine protease inhibitor PAI-1 to prevent, inhibit or
limit the onset of the physiological basis for the malady or condition in question.
10 The precise dosage to be employed depends upon several factors including
the host, whether in veterinary medicine or human medicine, the nature and severity
of the condition being treated, the mode of administration and the particular active
substance employed. The compounds may be administered by any conventional
route, in particular enterally, preferably orally in the form of tablets or capsules,
15 Administered compounds can be in the free form or pharmaceutically acceptable salt
forn as appropriate, for vise as a pharmaceutically particularly for use in the
myocardial infarction. arrhythmias, heart failure, kidney failure, stroke, peripheral
arterial occulsion, and related disease states). These measures will slow the rate of
20 progress of the disease state and assist the body in reversing the process direction in
a natural manner.
Any suitable carrier known to the art can be used to prepare the
pharmaceutical compositions. In such a composition, the carrier may be a solid,
25 liquid or mixture of a solid and a liquid. Solid compositions include powders, tablets
and capsules. A solid carrier can be one or more substances which may also act as
a flavoring agent, lubricant, solubilizer. suspending agent, binder, or tablet
disfritegrant. In powders, the carrier is a finely divided solid, which is in abmixture
with the finely divided active ingredient. In tablets the active ingredient is mixed with
30 a carrier having the necessary binding properties in suitable proportions and
compacted in the shape and size desired. Suitable solid carriers are magnesium
carbonate, magnesium slearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin,
tragacanth, methyl cellulose, hydroxymetriyl cellulose, sodium carboxymethyl
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cellulose, a low matting wax, cocoa butter, and the like. Encapsulating materials may
also be employed with the compounds of this invention, and the term "composition" is
intended to include the active ingredient in combination with an encapsulating
material as a formulation, wilh or without other carriers. Cachets may also be used in
5 the delivery of the anti-atherosclerotic medicament of this invention.
Sterile liquid compositions include solutions, suspensions, emulsions, syrups
and elixirs. The compounds of this invention may be dissolved or suspended in the
pharmaceutically acceptable carrier, such as sterile water, sterile organic solvent or a
10 mixture of both. Preferably the liquid carrier is one suitable for parental injection.
Where the compounds are sufficiently soluble they can be dissolved directly in
normal saline with or without the use of suitable organic solvents, such as propylene
glycol or polyethylene glycol. If desired, dispersions of the finely divided compounds
can be made-up in aqueous starch or sodium carboxymethyl cellulose solution, or in
15 a suitable oil, such as arachis oil. Liquid pharmaceutical compositions, which are
sterile solutions or suspensions, can be utilized by intramuscular, intraperitoneal or
subcutaneous injection. In many instances a liquid composition form may be used
instead of the preferred solid oral method of administration.
20 It is preferred to prepare unit dosage forms of the compounds for standard
administration regimens. In this way the composition can be subdivided readily into
striker doses at the physicians direction. For example, unit dosages may be made
up in packeted powders, vials or ampoules and preferably in capsule or tablet form.
The active compound present in these unit dosage forms of the composition may be
25 present in an amount of from about one gram to about fifteen grams or more, for
single or multiple daily administration, according to the particular need of the patient.
The daily dose of active compound will vary depending upon the route of
administration, the size, age and sex of the patient, the severity of the disease state,
and the response to the therapy as traced by blood analysis and the patients
30 recovery rate. By initialing the treatment regimen with a minimal daily dose of about
one gram, the blood levels of PAI-1 and the patients symptomatic relief analysis may
be used to determine whether a larger dose is indicated. Based upon the data
presented below, the projected daily dose for both human and veterinary use will be
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WO 2004/052893 PCT/US2003/038932
from about 25 to about 200 millgrams/kilogram per day, and more usually, from
about 50 to about 100 milligrams/kilogram per day.
The ability of the compounds of this invention to inhibit plasminogen activator
5 inhibitor-1 was established by the following experimental procedures.
PRIMARY SCREEN FOR THE PAI-1 INHIBITION
Test compounds were dissolved in DMSO at a final concentration of 10mM,
10 then diluted 100X in physiologic buffer. The inhibitory assay was initiated by the
addition of the lest compound (1-100 mM final concentration, maximum DMSO
concentration of 0.2%) in a pH 6.6 buffer containing 140 nM recombinant human
plasminogen activator inhibitor-1 (Molecular Innovations. Royal Oak, MI). Following
a 1 hour incubation at room temperature, 70 nM of recombinant human tissue
15 plasminogen activator (IPA) was added, and the combination at the test compound,
PA1-1 and tPA was incubated for an additional 30 minutes. Following the second
incubation. Spectrozyrne-tPA (American Diegnostica, Greenwich, CT). a
chromogenic substrate for tPA, was added and absorbance read at 405 nm at 0 and
60 minutes. Relative PAI-1 inhibition was equal to the residual tPA activity in the
20 presence of the test compound and PAl-1. Control treatments included the complete
inhibition of tPA by PAI-1 at the molar ratio employed (2:1), and the absence of any
affect of the test compound on tPA alone.
ASSAY FOR DETERMINING IC50 OF INHIBITION OF PAI-1
25
This assay is based upon the non-SDS dissociable interaction between tPA
and active PAI-1. Assay plates were initially coated with human tPA (10 mg/ml). Test
compounds were dissolved in DMSO at 10 mM. then diluted with physiologic buffer
(pH 7.5) to a final concentration of 1-50mM. Test compounds were, incubated with
30 human PAl-1 (50 ng/ml) for 15 minutes at room temperature. The lPA-coated plate
was washed with a solution of 0.05% Tweer 20 and 0.1% BSA, then the plate was
blocked with a solution of 3% BSA. An aliquot of the test compound/PAl-1 solution
was then added to the tPA-coated ptate, incubated at room temperature for 1 hour,
and washed. Active PAI-1 bound to the plate was assessed by adding an aliquot of a
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WO 2004/052893 PCT/US2003/038932
1:1000 dilution of the 33B8 monoclonal antibody against human PAl-1, and
incubating the plate at room temperature for 1 hour (Molecular Innovations, Royal
Oak Ml). The plate was again wasted, and a solution of goat anti-mouse lgG-
alkaline phosphatase conjugate is added at a 1:50,000 dilution in goat serum. The
5 plate was incubated 30 minutes at room temperature, washed, and a solution of
alkaline phosphatase substrate is added. The plate was incubated 45 minutes at
room temperature, and color development is determined at OD405nM. The quantitation
of active PAl-1 hound to tPA at varying concentrations of the test compound was
used to determine the IC50. Results were analyzed using a logarithmic best-fit
10 equation. The assay sensitivity was 5 ng/ml of human PAI-1 as determined from a
standard curve ranging from 0-100 ng/ml.

15
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WO 2004/052893 PCT/US2003/038932
Example 1
9-(4-Methylbenzyl)6-[4-(trifluoromethoxy)phenyl]-1,9-dihydropyrano[3,4-bi-
indole-3,4-dione
5 Step 1
EthyI 5-bromo-1- (4-methyIbenzyl)-1 H-indole-2-carboxyIate
NaH (60%, 3.88 g, 96.98 mmol) was added portion wise to a stirring solution of ethyl
5-bromo-1H-indole-2-carboxylate (20.0 g, 74.6 mmol) in DMF (140 mL) at 0ºC under
10 a nitrogen atmosphere over a period of 10 min. The mixture was then warmed to
room temperature. After the reaction mixture was stirred at room temperature for
one hour, 4-methylbenzyl bromide (14.2. g, 74 .8 mmol) was added and the mixrture
was stirred at room temperature overnight .The reaction was quenched with
aqueous ammonium chloride and diluted with water. The aqueous mixture was
15 extracted with ethyl acetate. The organic extract was washed with water and brine,
dried over anhydrous. magnesium sulfate. This mixture was concentrated to give a
semi-solid contained 0.8 mote equivalent DMF. Mass spectrum (ESI, (M+H)*) m/z
372. 1H NMR (400 MHz, DMSO-d6) d 7.93 (d, 1H. J = 7.94 Hz), 7.55 (d, 1H, J =
9.01 Hz), 7.40 (dd. 1H, J= 8.85 and 1.98 Hz), 7.32 (s, 1H), 7.04(d, 2H, J= 7.96 Hz),
20 6.90 (d, 2H, J = 7.94 Hz), 5.79 (s, 2H), 4.28 (q, 2H), 2.20 (s, 3H). and 1.28 ppm (t,
3H).
Step 2
Ethyl 1 -(4-methyIbenzyl)-5-[4-(trifluoromethoxy)phenyI]-1H-indole-2-
25 carboxylate
A mixture of ethyl 5-bromo-1-(4-methylbenzyl)-1H-indol-2-carboxylate (5.8 g, 15.54
mmol), 4-(trifluoromethoxy)phenylboronic acid (8.2 g. 38.85 mmol), potassium
carbonate (5.4 g. 38.85 mmol), (1,1-bis(diphenylphosphiro)-ferrocene]dichloro-
30 palladium (lI) complex with methylene (1:1) (4.41 g, 5.44 mmol) in dioxane-water
(10:1, 154 mL) was stirred at 70ºC for two days. The reaction mixture was diluted
with water and extracted with ethyl acetate. The organic extract was washed with
water and brine, and then concentrated to an oil. This residue was crystalized from
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WO 2004/052893 PCT/US2003/038932
ethyl ether to afford the title compound as an off-white solid, m.p, 77-78°C. Mass
spectrum (ESI, [M+H)+) m/z454. 1H NMR (400 MHz; DMSO-d6) d 8.00 (s, 1H), 7.78
(d, 2H, J = 3.70 Hz). 7.67 (d, 1H, J = 8.85 Hz), 7.61 (dd, 1H, J = 8.70 and 1.66 Hz),
7.44 (s, 1H), 7.42-7.41 (m, 3H). 7.06 (d, 2H. J = 7.94 Hz) 6.04 (d, 2B, J = 8.09Hz)
5 5.83 (s, 2H), 4.29 (q, 2H), 2.21 (s, 3H), and 1.30 ppm (t. 3H).
Elemental Analysis for C26H22F3NO5:
Calculated: C, 68.87; H, 4.89; N, 3.09.
Found: C. 69,00; H. 4.66; N, 3.06.
10 Step 3
{1-(4-Methylbensyl)-5-(4-(trifluromethoxy)phenyl]-1H-indol[2-yl}methanol
Lithium aluminum hydride (0.244 g, 6.1 mmol) was added portionwise to a stirring
15 carboxylate (2.0 g, 4.4 mmol) in ethyl ether (17 mL) al 0ºC under a nitrogen
atmosphere over a period of 6 minutes. The mixture was then warmed up to room
temperature. After the reaction mixture was stirred at room temperature for 5 hours,
the reaction was carefully quenched with water and filtered. The filtrate was
extracted with ethyl acetate. The organic extract was washed with water and brine;
20 dried over anhydrous magnesiun sulfate and evaporated to afford the title compound
as a while solid (1.56 g. B6 %). Mass spectrum (ESI, [M+H]+) m/z 412. 1HNMR
(400 MHz, DMSO-d6); d 7.81 (s, 1H). 7.76 (d. 2H, J = 6.71 Hz), 7.45-7.35 (m, 4H),
7.08(d,2H,J=7.74Hz).6.96(d,2H.J=8.09Hz),6.50(s,1H),5.44(s,2H),5.35(t
1H), 4.60 (d. 2H), and 2.23 ppm (t, 3H).
25 Elemental Analysis for C24H20F3NO2:
Calculated; C, 70.07; H, 4.90; N, 3.40.
Found; C. 69.30; H, 4.75; N, 3.34.
Step 4
30 {1-(4-Methylbenzyl)-5-[4-(trifluoromethoxy)pheny]-1-H-lndol-2-yl}methyl acetate
Acetyl chloride (0.222 mL, 3.08 mmol) was added to a stirring solution of (1-(4-
methylbenzyl)-5-14-(trifluoromethoxy)phenyl]-1H-indol-2-yl}methanol (0.507 g. 1.23
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WO 2004/052893 PCT/US2003/038932
mmol) and N,N-diisopropylethylamine (0.547 mL, 3.08 mmol) in methylene chloride
(8 mL) at 0 ºC under a nitrogen atmosphere over a period of 5 minutes. After the
reaction mixture was stirred at room temperature overnight, the reaction was
quenched carefully with water. The aqueous mixture was extracted with ethyl
5 acetate. The organic extract was washed with water and brine, dried over anhydrous
magnesium sulfate and evaporated to afford the title compound as a solid (0.557 g.
99.6 %). mp: 125-126 ºC. Mass spectrum (ESI, [M+H]+) m/z 454. 1HNMR (400
MHz, DMSO-d6): d 7.86 (s, 1H). 3.77 (d, 2H, J = 8.96 Hz), 7.48-7.40 (m, 4H). 7.09 (d.
2H, J = 7.94 Hz). 6.90 (d, 2H, J = 7.94 Hz), 6.70 (s, 1H), 5.44 (s. 2H), 5.23 (s, 2H).
10 2.23 (s, 3H), and 1 .85 ppm (s, 3H).
Elemental Analysis for C26H22F3NO3;
Calculated: C. 68.32; H. 4.94.; N. 3.07.
Found: C, 67.96; H. 4.57; N, 2.96.
15 Step 5
{2-[(Acetyloxy)methyl]-1 -[4-methylbenzyI) -5-[4-[trifluoromethoxy)phenyI]-1H-
indol-3-yl}(oxo)acetic acid
Oxalyl chloride (1.05 mL) 'was added dropwise to a stirring solution of {1-(4-
20 methylbenzyl)-5-[4-(trifluoromethoxy)phenyI]-1H-indol-2-yl}methyl acetate(0.515g)in
THF (17 mL) at room temperature over a period of 5 minutes under a nitrogen
atmosphere. After the reaction mixture Was stirred at room temperature for 4 hours
the reaction was quenched carefully with water. The aqueous mixture was extracted
with ethyl acetate. The extract was washed with water, and brine, dried over
25 anhydrous magnesium sulfate, and concentrated to give the title compound as a light
brown solid, mp: 79-30 ºC. Mass spectrum (ESI, [M-H]") m/z 524. 1HNMR (400
MHz, DMSO-d6): d 14.50 (br s. 1H), 8.20 (s, 1H), 7.74 (d, 2H, J = 8.65 Hz). 7.70 (d,
1H. J = 8.69 Hz) 7.62 (d, 1H, J = 8.55 Hz), 7.47 (d, 2H, J = 8.25 Hz). 7.15 (d, 2H, J =
7.33 Hz), 6.93 (d. 2H, J= 7.78 Hz), 5.64 (e, 2H), 5.53 (s, 2H), 2.25 (s, 3H), and 1.83
Elemental Analysis for C28H22F3NO5:
Calculated: C. 64.00; H. 2.67; N. 2.67.
Found: C. 63.77; H, 3.99; N. 2.65.
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WO 2004/052893 PCT/US2003/038932
Step 6
{2-(Hydroxymethyl)-1-(4-methyIbenzyl)-5-[4-(trifluoromethoxy)phenyI]-1H-indol-
3-yl}(oxo)acetic acid, potassium salt
5 A solution of {2-[(acetyloxy)methyl]-1-(4-methylbenzyl)-5-[4-(trifluoromethoxy)phenyl]-
1H-indol-3-yl}(oxo)acetic acid (0.50 g, 0.96 mmol) and aqueous potassium hydroxide
(1.0 N. 2.38 mL, 2.38 mmol) In THF : water (1 : 1, 16 mL) was stirred at room
temperature for 4 h. The reaction mixture was evaporated to dryness. The residue
solid was stirred in water : hexane (8 : 92. 100 mL) and filtered to give the title
10 compound as a white solid (0.4 g), mp: 248-249.5 °C. Mass spectrum (ESI, [M-H]')
m/z 482. 1HNMR (400 MHz, DMSO-d6). d 8.51 (e. 1H) 7.73 (d, 2H, J =6.86 Hz).
7.50 (d, 1H. J = 8.56 Hz), 7.45 (d, 1H. J = 8.55 Hz), 7.43 (d, 2H. J = 7.94 Hz), 7.09
(d, 2H. J = 8.09 Hz), 7.05 (d, 3H, J= 8.09 Hz). 6.74 (t, 1H). 5.55 (s, 2H), 4.80 (d, 2H,
J=6.80 Hz), and 2.23 ppm (s, 3H).
15 Elemental Analysis for C26H20F3NO5 1.0 K 0.8 H2O:
Calculated: C, 58.27", H, 3.87; N. 2.61.
Found; C, 58.14; H. 3.82; N. 2.59.
Step7
20 9-(4-Methylbenzyl)-6-[4-(trifluoromethoxy)phenyl]-1,9-dihydropyrano[3,4-b]-
indole-3,4-dione
{2-{Hydroxymethyl)-1-(4-methylbenzyl)-5-[4-trifluoromethoxy)phenyl]1-H-indol-3-yl)-
{oxo} acetic acid, potassium salt (0.355 g) was partioned between methylene
25 chloride (400 mL) and 15 % aqueous HCl (100 mL) with stirring. The organic layer
was separated, washed with water and evaporated to afford the title compound as an
off-white solid (0.304 g), mp: 243-244ºC. Mass spectrum (ESI, [M-H]-) m/z 464.
1HNMR (400 MHz, DMSO-d6): d 8.23 (s, 1H), 7.81 (d, 2H, J = 7.49 Hz), 7.77 (d, 1H,
J= 8.40 Hz), 7.67 (d. 1H, J = 8.71 Hz),7.46 (d, 2H, J = 7.94 Hz) 7.16 (s,4H) 5.92
30 (s.2H),5.50(s,2H),and 2.25ppm(s,3H).
Elemental Analyses for C20B16F3NO4:
Calculated: C. 67.10; H, 3.90; N, 3.01.
Found: C, 66.32; H. 3.71: N, 2.91.
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WO 2004/052893 PCT/US2003/038932
Example 2
9-Benzyl-6-[4-(trifluoromethoxy]phenyl]-1,9-dihydropyrano[3,4-b]incdole-3,4-
dione
5
Step1
Ethyl 5-bromo-1-benzyl-1H-indole-2-carboxylate
The title compound was prepared from Ethyl 5-bromo-1H-indole-2-carboxylate and
10 benzyl bromide in substantially the same manner, as described in step 1 of Example
1. The product was obtained as a light yellow solid. Mass spectrum (ESI, [M+H]+}
m/z 358. 1H NMR (300 MHz, DMSO-d6) d 7.94 (s. 1H). 7.55 (d, 1H, J =9.01 Hz),
7.42 (d. 1H, J = 8.85 Hz), 7.34 (s, 1H), 7.30-7.21 (m, 3H). 6.99 (d, 2H, J = 7.94 Hz),
5.65 (s, 2H), 4.23 (q. 2H). and 1.28 ppm (t, 3H).
15 Elemental Analysis for C16H16BrNO2:
Calculated: C, 60.35; H, 4.50; N, 3.91.
Found; G, 69.19: H. 4.51; N, 3.75.
Step2
20 Ethyl1-benzyl-5-[4-(trifluoromethoxy)phenyl]-1H-indole-2-carboxylate
The title compound was prepared from ethyl 5-bromo-1-benzyl-1H-indole-2-
carboxylate and 4-(trifluoromethoxy)phenylboronic acid in substantially the same
manner as described in step 2 of Exanmple 1. The product was obtained as an oil.
25 Mass spectrum (ESI, [M+H]+) m/z 440. 1H NMR (400 MHz, DMSO-d6) d 8.01 is.
1H). 779 (d, 2H, J = 8.86 Hz) 7.57 (d, 1H, J = 8.85 Hz), 7.62 (dd. 1H, J= 8.70 and
1.68 Hz), 7.44-7.42 (m, 2H), 7.28-7.20 (m, 3H), 7.04 (d, 2H, J = 794 Hz), 5.88 (s,
2H), 4.28 (q, 2H), 1.29 (t, 3H).
30 Step 3
{1 benzyl-5-[4-(trifluoromethoxy]=1H-indol-2-yl}methanol
The title compound was prepared from ethyl 1-benzyl-5-[4-(trifluoromethoxy)phenyl}-
1H-indole-2-carboxylate (step 3 of Example 1) and lithium aluminum in substantially
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WO 2004/052893 PCT/US2003/038932
the same manner, as described in step 1 of Example 21. The product was obtained
as a white solid, mp: 108-109 ºC. Mass spectrum (ESI, [M+H]+) m/z 398. 1H NMR
(400 MHz, DMSO-d6) d 7.82 (s. W), 7.76 (d. 2H. J = 8.86 Hz), 7.42-7.36 (m, 4H),
7.29 (d, 1H, J= 7.03 Hz), 7.27 (d, 1H, J = 7.63 Hz), 7.23-7.20 (m, 1Hz). 7.08 (d, 2H,
5 J = 7.03 Hz), 6.52 (s, 1H), 5.50 (s, 2H), 5.36 (1, 1H), and 4.60 ppm (d, 2H).
Elemental Analysis for C23H18F3NO2:
Calculated: G, 69.52; H, 4.57; N, 3.52.
Found: C, 68.21; H, 4.38; N. 3.40.
10 Step 4
{1-benzyl-5-[4-(trifluoromethoxy)phenyl]-1H-indol-2-yl}methyI acetate
The title compound was prepared from {1-benzyl-5-[4-(trifluoromethoxy)phenyl]-1H-
indol-2-yl}methanol and acetyl chloride in substantially the same manner as
15 described in step 4 of Example 1. The product was obtained as an oil. Mass
spectrum (ESI. [M+H]+)m/z 440. 1H NMR (300 MHz. DMSO-d6) d 7.88 (s, 1H), 7.77
(d, 2H, J= 8.87 Hz), 7.43-7.40 (m. 4H), 7.29-7,22 (m, 3H). 7.00 (d, 2H, J= 7.01 Hz),
6.72 (s, 1H), 5.50 (s, 2H). 5.24 (s. 2H), and 1.30 ppm (s, 3H).
Elemental Analysis for C25H20F3NO3.
20 Calculated: C, 68.33; H,4.59; N. 3.19.
Found: C, 68.19; H. 4.70; N, 3.08.
Step5
{2-[(Acetyloxylmethyl]-1-benzyl-5-[4-(trifluoromethoxy)phenyl]-1H-indol-3-yl]-
25 (oxo)acetic acid
The title compound was prepared from {1-benzyl-5-[4-(trifluoromethoxy)phenyI]-1H-
indol-2-yl]methyl acetate and oxalyl chloride in substantially the same manner, as
described in step 5 of Example 1. The product was obtained as a brown solid; mp:
30 85-86 ºC. Mass spectrum (ESI, [M-H]-) m/z 510, 1H NMR (400 MHz, DMSO-d6) d
14.40 (br s. 1H), 6.20 (s, 1H), 7.76 (d. 2H, J = 6.72 Hz), 7.72 (d, 1H, J = 8.70 Hz).
7.63 (d. 1H, J= 3.70 Hz), 7.43 (d, 2H, J= 8.24 Hz), 7.34-7.25(m.3H),7.08(d, 1H, J =
7.23 Hz), 5.70 (s. 2H), 5.53 (s, 2H), and 1.78 ppm (s, 3H).
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WO 2004/052893 PCT/US2003/038932
Elemental Analysis for C27H20F3NO6:
Calculated: C, 63.41; H, 3.94; N. 2.74.
Found: C, 63.02; H, 3.97; N, 2.64.
5 Step 6
{2-(Hydroxymethyl)-1-benzyl-5-[4-(trifIuoromethoxy)phenyI]-1H-indoI-3 -yl}-
(oxo) acetic acid, potassium salt
The title compound was prepared from {2-(acetyloxy)methyl]-1-benzyl-5-[4-(trifluoro-
10 methoxy)phenyl]-1H-indol-3-yl)(oxo)acetic acid and aqueous potassium hydroxide in
substantially the same manner, as described in step 6 of Example 1. The product
was obtained as a white solid; mp: 280-282ºC. Mass spectrum (ESI, [M-H]-) m/z468.
1H NMR (400 MHz, DMSO-d6) d 3.51 (s, 1H), 7.72 (d. 2H. J = 8.70 Hz), 7.52 (d, 1H.
J = 3.55 Hz), 7.47-7.43 (m. 3H), 7.31-23 (m, 2H). 7.23 (d, 1H. J = 7.16 Hz), 7.15 (dr
15 2H, J = 7.1B Hz), 5.75 (i, 1 H), 5.63 (s, 2H). and 4.61 ppm (d. 2H. J = 6.72 Hz).
Calculated: C, 57.14; H, 3.64; N. 2.67.
Found: C, 57.05; H, 3.42; N, 2.55.
20 Step 7
9 -Benzyl-6-[4-(trifluoromethoxy)phenyl]-1,9-dihydropyrano[3,4-b]indoIe-3,4-
dione
The title compound was prepared from {2-(Hydroxymethyl)-1-benzyl-5-[4-(trifluoro-
25 methoxy)phenyl]-1H-indol-3-yl}(oxo)acetic acid, potassium salt and aqueous HCI in
substantially the same manner, as described in step 7 of Example 1. The product
was obtained as an off-white solid; mp: 239-240ºC. Mass spectrum (ESI. [M+H]+)
m/z 452. 1H NMR (400 MHz, DMSO-d6) d 6.24 (s, 1H), 7.81 (dd, 2H, J = 6.86 and
2.14 Hz), 7.74 (d, 1H, J= 8.56 Hz), 7.68 (dd, 1H, J= 6.54 and 1.83 Hz), 7.46 (d, 2H,
30 J = 7.54 Hz), 7.37-7.26 (m, 6H,), 5.93 (s, 2H). and 5.56 ppm (s, 2H).
Elemental Analysis for C25H10F3NO4:
Calculated: C, 68.52: H. 3.57; N, 3.10.
Found: C, 69.53; H, 3.60; N, 3.04.
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WO 2004/052893 PCT/US2003/038932
9-(4-Methylbenzyl)-8-(3-methylphenyl)-1,9-dihydropyranol[3,4-b]indol-3,4-dione
5 Step 1
Ethyl 1 -(4-methylbenzyl)-5-(3-methylphehyl)-1H-indole-2-carboxylate
The.title compound was prepared from ethyl 5-bromo-1 -(4-methylbenzyl-1 H-indole-
2-carboxylate (step 1 of Example 1) and 3-methylphenylboronic acid in substantially
the same manner, as described in step 2 of Example 1. The product was obtained
10 as a solid. Mass Spectrum (ESI, [M+H]+) M/z 384. 1H NMR (400 MHz, DMSO-d6) d
7.95(s, 1H),7.63 (d, 1H, J= 7.70 Hz), 7.60 (d, 1H, J = 8.47 Hz) 7.43 (s, 1H),7.44
(d, 1H. J = 7.95 Hz), 7.39 (e, 1H), 7.33 (t, 1H, J = 7.63 Hz), 7.14 (d. 1H. J = 7.49 Hz).
7.07 (d, 2H, J= 7.94 Hz), 6.93 (d, 2H. J = 7.94 Hz). 6.33 (s, 2H). 4.29 (q, 2H, J=
7.17 Hz), 2.37 (s, 3H), 2.21 (s. 3H), and 1.30 ppm (t, 3H, J = 7.18 Hz).
15
Step2
[1 -(4-methylbenzyl)-5-(3-methylphenyl)-1H-indol-2 -yl)methanol
The title compound was prepared from ethyl 1-(4-methylbenzyl)-5-(3-methylphenyl)-
1H-indole-2-carboxylate and lithium aluminium in substantially the same manner. as
20 described in step 3 of Example 1. The product was obtained as a white solid. Mass
spectmm (ESI, [M+H]+) m/z342. 1H NMR (400 MHz, DMSO-d6) d 7.77 (s, 1H), 7.46
(s. 1H). 7 42 (d, 1H, J= 7.79 Hz). 7.37 (d, 1H, J= 8.55 Hz), 7.33 (d, 1H, J= 8.55 Hz).
7.31 (d, 1H, J= 7.02 Hz), 7.29 (d. 1H, J= 7.48 HZ), 7.10-7.07 (m, 4H), 6.96 (d, 2H,
J =7.94 Hz), 6.48 (s. 1H), 5.54 (s. 2H), 5.33 (s. 1H), 4.60 (s. 2H). 2.36 (s, 3H), and
Elemental Analysis for C24H22NO- O.5 H2O:
Calculated: C, 82.25: H, 6.90; N, 4.00.
Found: C. 82.05; H. 6.93; N, 3.86.
30 Step 3
[1-{4-Methylbenzyl}-5-(3-methylphenyl)-1H-indol-2-yl]methyl acetate
The title compound was prepared from [1-(4-methylbenzyl)-5-(3-methylphenyl)-1H-
indol-2-yl]methanol and acetyl chloride in substantially the same manner, as
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WO 2004/052893 PCT/US2003/038932
described in step 4 of Example 1. The product was obtained as a brown oil. Mass
spectrum [ESI. [M+H]+) m/z 334. 1H NMR (400 MHz. DMSO-d6) d 7.82 (s, 1H), 7.47
(s. 1H). 7.45-7.40 (m, 3H). 7.31 (t, 1H ,J = 7.74 Hz), 7.13-7.08 (m. 3H), 6.90 (d, 2H, J
= 3.09 Hz). 6.63 (s, 1H), 5.43 (s, 2H). 5.22 (s, 2H), 2.36 (s, 3H). 2.23 (s, 3H), and
5 1.85 ppm(s,3H).
Elemental Analysis for C26H25NO2:
Calculated: C. 81.43: H,6.57; N, 3.65.
Found: C, 81.75; H. 6.67; N, 3.25.
10 Step 4
[2-[(Acetyloxy)methyl]-1-[4-methylbenzyl)-5-(3-methylphenyl)-1H-indol-3-yl]-
(oxo) acetic acid
The title compound was prepared from [1-(4-methylbenzyl)-5-(3-methylphenyl)-1H-
15 indol-yl]methyl acetate and oxalyl chloride in substantially the same manner, as
described in step 5 of Example 1. The product was obtained as a brown solid; mp:
89-9CºC. Mass spectrum (ESI, [M-H]-) m/z 454. 1H NMR (400 MHz, DMSO-d6) d
8.18 (s, 1H), 7.57 (d, 1H, J= 8.69 Hz), 7.53 (dd. 1H, J= 7.17 and 1.53 Hz), 7.44-7.41
(m. 2H), ), 7.36 (i, 1H. J= 7.43 Hz), 7.18 (d, 1H, J = 7.23 Hz), 7.13 (d. 1H, J = 7.94
20 Hz), 5.63 (s, 2H), 5.53 (s. 2H), 2.39 (s, 3H), 2.24 (S. 3H). and 1.83 ppm (s, 3H).
Elemental Analysis for C26H25NO5-0.5 H2O:
Calculated: C. 72.40; H. 5.04; N, 3.02.
Found: C, 72.41; H, 5.44; N. 2.96.
25 Step 5
9-(4-Methylbenzyl)-6-(3-methyIphenyl)-1,9-dihydropyrano[3,4-b]indole-3,4-dione
The title compound was prepared from [2-(acetyloxy)methyl]-1-(4-methylbenzyl)-5-
(3-methylphenyl)-1H-indol-3-yl](oxo)acetc acid and aqueous potassium hydroxide in
30 substantially the same manner, as described in step 6 of Example 1 followed by the
treatment with aqueous HCl in substantially the same manner, as described in step 7
of Example 1. The product was obtained as a white solid; mp: 216-217ºC. Mass
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WO 2004/052893 PCT/US2003/038932
spectrum (ESI. [M+H]+) m/z396. 1H NMR (400 MHz, DMSO-d6) d 8.21 (s, 1H), 7.71
(d, 1H, J= 3.56 Hz), 7.64 (dd, 1H. J= 8.75 and 1.23 Hz). 7.49 (s, 1H), 7.46 (d. 1H J
= 3.09 Hz), 7.36 (t, 1H. J = 7.63 Hz). 7.19-7.14 (m, 5H), 5.91 (s, 3H). and 5.49 (s.
2H), 2.39 (s, 3H), 2.26 ppm (s, 3H).
5 Etemental Analysis for C26H21NO3- 0.4 H2O;
Calculated: C,77.56; H, 5.46; N, 3.48.
Found. C. 77.41, H. 5.46, N, 3.49.
Example 4
10 9-(4-tert-butyIbenzyl)-6-(3-methylphenyI)-1,9-dihydropyrano[3,4-b]indole-3,4-
dione
Step1
(5-Bromo-1H-indol-2-yl)methanol
15
The tilte compound was prepared from ethyl 5-bromo-1H-indole-2-carboxylate and
lithium aluminum in substantially the same manner, as described in step 3 of
Example 1. The product was obtained as a solid; mp: 111-112ºC. Mass spectrum
(ESI, [M-H]-) m/z 224. 1H NMR (400 MHz, DMSO-d6) d 7.62 (s, 1H) 7.27 (d, 1H J =
20 8.02 Hz), 7.12 (d, IH, J = 8.02 Hz), 6.25 (s, 1H), 5.29 (t. 1H, J = 5.50 Hz), 4.59 ppm
(d.2H. J=5.65Hz).
Elemental Analysis for C9H8BrNO;
Calculated: C, 47.62; H, 3,57; N, 6.20.
Found: C, 47.94; H, 3,42; N, 6.20.
35
Step 2
[5 -(3-MethylphenyI)-1H-indol-2-yI]methyI acetate
A mixture of (5-bromo-1H-indol-2-yl) methanol (9 g, 39.3 mmol). 3-methylbenzene-
30 boronic acid (6.14 g, 43.78 mmol), potassium carbonate (13.75 g, 90.5 mmol),
palladium(ll) acetate (0.045 g) and tetrabutylammonium bromide (12.84 g, 39.6
mmol) In 10 % dioxane in water (degassed. 0.36 L) was stirred at 70 ºC. The
reaction was monitored by TLC. Additional 3-methylbenzeneboronic acid (5.6 g,39.8
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WO 2004/052893 PCT/US2003/038932
mmol) was added. After no 5-bromo-1-(4-tert-butylbenzyl)-1H-indole was detected
by TLC, the reaction was cooled lto room temperature and the solvent was decanted.
The residual thick oil was stirred with hexane. The hexane was decanted and the
residue was partitioned between water and ethyl acetate. The ethyl acetate layer
5 was separated, crashed with water filtered and concentrated. The, residue was
purified by flash column chromatography using hexane/ethyl acetate (55 : 45) as an
eluant to give [5-(3-Methylphenyl)-1H-indol-2-yl]methanol as a brown solid (6.0 g).
Reaction of [5-(3-Methylphenyl)-1H-indol-2-yl]methanol with acetyl chloride according
to the procedure described in step 4 of Example 1 afforded the title compound as an
10 oil (3.94 g); Mass spectrum (ESI. [M-H]-) m/z 278. 1H NMR (400 MHz. DMSO-d6) d
7.76 (s, 1H), 7.46 (s, 1H), 7.42-7.36 (m, 3H). 7.30(s, 1H), 7.10 (d, 1H, J=7.48Hz),
6.50 (s. 1H), 2.37 (s, 3H), and 2.07 ppm (s, 3H).
Step3
15 [1-(4-tert-Butylbenzyl)-5-(3-methylphenyl)-1H-indol-2-yl]methyl acetate
Potassium carbonate (0.3 g, 2.15 mmol) was added to a stirring solution of 5-(3-
methylphenyl)-1H-indol-2-yI]methyI acetate (0.5 g, 1.79 mmol) in DMF (5 mL) at room
temperature under a nitrogen atmosphere. 4-tert-Butylbenzyl bromide
mmol)was added and the mixture was stirred at room temperature overnight. The
30 reaction was monitored by TLC. An additional amount of 4-tert-butylbenzyl bromide
(0.34 g, 1.8 mmol) was added and stirring continued at room temperature for three
days. The reaction was quenched with aqueous ammonium chloride. diluted with
water and extracted with eithyl acetate. The organic extract was washed with water
and brine, dried over anhydrous magnesium sulfate and evaporated to dryness. The
25 residue was purified by flash column chromatography using hexane/ethyl acetate
(85;15) as an elvant to give the title compound as a light brown oil (0.284g). Mass
spectrum (ESI. [M+H]+) m/z 426. 1H NMR (400 MHz, DMSO-d6) d 7.83 (s. 1H),
7.43-7.47 (m,2H), 7.44-7.41 (m, 2H), 7.32-7.29 (m, 3H) 7.11 (d, 1H, J = 7.49 Hz),
8.31 (d, 2H, J = 8.24 Hz), 6.69 (s, 1H), 5.44 (s, 2H), 5.23 (s, 2H), 2.36(s, 3H), 1.78
30 (s, 3 H), and 1.21 ppm (s. 9H).
Elemental Analysis for C29H31NO2- 0.3 H2O:
Found: C, 80.82; H, 7.39; M, 3.25,
Calculated: C,80.90;H, 7.45; M. 3.17.
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WO 2004/052893 PCT/US2003/038932
Step 4
[2-[(Acetyloxy)methyI]-1-(4-tert-butyIbenzyl)-6-(3-methylphenyI)-1H-indol-3-yI]-
(oxo)acetic acid
5 The title compound was prepared from [1-(4-tert-butylbenzyl)5-(3-methylphenyl)-1H
indol-2-yl)methyl acetate and oxalyl chloride in substantially the same manner, as
described in step 5 of Example 1. The product was obtained as a light brown solid;
mp: 91-92°C. Mass spectrum (ESI, [M-H]-) m/z496. 1H NMR (400 MHz, DMSO-d6)
d 8.18 (s, 1H), 7.72 (d, 2H, J = 6.71 Hz), 7.61 (dd, 1H. J= 8.70 and 1.38 Hz), 7.45-
10 7.41 (m, 2H), 7.38-7.33 (m, 3H), 7.16 (d, 1H, J = 3.70 Hz), 7.00 (d. 2H, J= 3.24 Hz),
5.64 (s, 2H), 5,53 (s, 2H), 2.39 (s, 3H). 1.76 (s, 3H), and 1.22 ppm (s, 9H).
Elemental Analysis for C31H31NO5- 0.5 H2O:
Calculated: C, 73.60, H, 6.37; N, 2.77.
Founds C. 73.43; H, 8,57; N. 2.84.
15
Step5
9-(4-tert-Butylbenzyl)-6-(3-methylphenyl)-1,9-dihydropyrano[3,4-b]indole-3,4-
dione
20 A solution of [2-{(acetyloxy)methyl]-1-(4-tert-butylbenzyl)-5-(3-methylphenyl)-1H-
lndol-3-yl](oxo)acetic acid (0.27 g, 0.63 mmol) and aqueous potassium hydroxide
(1.0 N, 1.1 mL 1.1 mmol) in THF:MeOH (1:1, 6.6 mL) was stirred at room
temperature for 4 h. The reaction mixture was evaporated to dryncss. The residue
was washed with water and hexane to give an oil. This oil was partitioned between
25 methylene chloride (60 mL) and 15 % aqueous HCl (15 mL) with stirring. The
organic layer was separated, washed with water and evaporated to aftord the title
compound as an off-white solid, mp; 196-197ºC. Mass spectrum (ESI, [M+H]+) m/z
438. 1HNMR (400 MHZ, DMSO-d6): d 6.21 (s, 1H), 7.75 (d. 2H, J = 8.55 Hz), 7.66
(dd, 1H, J = 8.70 and 1.52 Hz), 7.50 (s, 1H), 7,47 (d, 1H. J = 8.71 Hz), 7.38-7.35 (m,
30 3H), 7.20-7.17 (m. 3H), 5.92 (s, 2H). 5.50 (s. 2H), and 2.39 ppm (s, 3H).
Elemental Analysis for C28H27NO3- 0.5 H2O;
Calculated: C. 76.64; H, 6.26: N, 3.16
Found: C, 78.54; H. 6.46; N. 3.04
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WO 2004/052893 PCT/US2003/038932
Example 5
6-[Benzyloxy)-9-(4-methylbenzyl)-1,9-dihydropyrano]3,4-b]indol-3, 4-dione
5 Step 1
[5-(Benzyloxy)-1H-indol-2-yl]methanol
The title compound was prepared from ethyl 5-(benzyloxy)-1H-idole-2-carboxylate
and lithium aluminum hydride according to the procedure described in step 3 of
10 Example 1. The product was obtained as a white solid, mp: 106-107ºC. Mass
spectrum (ESI, [M+H]+) m/z 254. 1H NMR (400 MHz. DMSO-d6) d 10.80 (s, 1H)
7.45 (d, 2H. J = 7.33 Hz). 7.37 (t. 2H, J = 7.49 Hz), 7.30 (d. 1H. J = 7.18 Hz), 7.20 (d,
1H, J = 8.71 Hz), 7.04 (s. 1H). 6.74 (dd, 1H, J = 6.70 and 1.46 Hz). 6.1 S (s, 1H), 5.16
(t, 1H, J = 5.49 Hz), 5.01 (s. 3H), 4.55 ppm (d, 2H, J= 5.50 Hz).
15 Elemental Analysis for C15H15NO4:
Calculated: C. 75.87; H, 5.97; N. 5.53.
Found: C, 75.95; H, 6.11; N. 5.41.
20 [5-(Benzyloxy)-1H-indol-2-yl]methyl acetate
The title compound was prepared from [5-(benzyloxy)-1H-indol-2-yl]methanol and
acetyl chloride in substantially the same manner as described in step 4 of example
1, The product was obtained as a gray solid. Mass spectrum (ESI, [M+H]+) m/z 296,
25 1H NMR (400 MHz, DMSO-d6) d11.03 (s,1H), 7.45 (d, 2H, J = 7.33 Hz), 7.38 (t, 2H,
J = 7.49 Hz), 7.30 (d, 1H, J = 7.20 Hz), 7.24 (6, 1H, J = 8.71 Hz), 7.08 (s, 1H), 6.61
(dd, 1H, J= 8.70 and 1.45 Hz). 6.34 (s, 1H). 5.13 (s. 2H), 5.06 (s, 2H). 2.04 ppm (s,
3H).
Elemental Analysis for C10H17NO3- 0.2 H20:
30 Calculated: C. 72.32; H, 5.67; N, 4.69,
Found; C, 72.21; H. 5.75; N, 4.66.
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WO 2004/052893 PCT/US2003/038932
Step 3
{5-(Benzyloxy)-1-(4-methylbenzyl)-1H-indol-2-yl}methyI acetate
The title compound was prepared from [5-(benzyloxy)-1H-indol-2-yl)methyl acetate
5 and 4-methylbenzyl bromide in substantially the same manner, as descried in step 3
of Example 4. The product was obtained as a white solid. Mass spectrum (ESI,
[M+H]+) m/z400. 1H NMR(300 MHz, DMSO-d6)d7.45(d.2H, J= 8.34 Hz), 7.37(t,
2H, J=7.49 Hz), 7.31 (d, 1H. J= 7.30 Hz). 7.26 (d, 1H, J= 7.56 Hz).7.15 (d, 1H, J=
2.44 Hz), 7.07 (d, 2H, J =8.76 Hz), 6.65 (d, 2H, J = 8.81 Hz), 6.84 (s, 1H). 6.25 (s,
10 1H), 5.35 (s, 2H). 5.17 (s, 2H),5.09 (s, 2H) 2.22 (s, 3H), 1.84 ppm (s, 3H).
Elemental Analysis for C20H23NO3:
Calculated: C. 78.17; H, 6.31. N, 3.51.
Found: C. 78.01; H. 6.35; N, 3.46.
15 Step 4
[2-[(Acetyloxy)methyl]-5-(benzyloxy)-1-(4-methylbenzyl)-1H-indol.3-yl](oxo)-
acetic acid
The title compound was prepared from [5-(benzyloxy)-1-(4methylbenzyl)-1H-indol-2-
20 yl]methyl acetate and oxalyl chloride in substantially the same manner, as described
in step 5 of Example 1. The product was obtained as an off-white solid; mp: 91-92ºC,
Mass,spectrum (ESI, [M-H]-) m/z 470. 1H NMR (400 MHz. DMSO-d6) d 7.61 (s, 1H),
7.50 (s. 1H).7.47 (d,2H, J = 8.24Hz),7.39 (t,2H. J = 7.84Hz).7.32 (d, 1H, J =7.02
Hz) 7.11 (d. 2H, J = 8.10 Hz), 7.03 (d. 1H, J = 3.99 Hz), 6.95 (d. 2H, J = 7.79 Hz),
25 5.54 (s, 2H), 5.46 (s, 2H), 5.10 (s, 2H), 2.23 (s, 3H), 1.82 ppm (s, 3H).
Elemental Analysis for C25H25NO5* 0.6 H2O:
Calculated: C, 69.73; H, 5.48; N, 2.90.
Found: C, 69.60; H, 5.30; N, 2.80,
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WO 2004/052893 PCT/US2003/038932
Step5
6-(Benzyloxy)-9-(4-methylbenzyl)-1,9-dihydropyrano[3,4-d]indole-3,4-dione
The title compound was prepared from [2-[(acetyloxy)methyl]-5-(benzyloxy)-1-(4-
5 methylbenzyl)-1H-indol-3yl](oxo)acetic acid and aqueous potassium hydrooxide
according to the procedure described in step 6 of Example 1, followed by the
treatment with aqueous HCl in substantially the same manner as described in step 7
of Example 7. The product was obtained as an off-white solid; mp: 206-207ºC. Mass
spectrum (ESI, [M+H]+)m/z412. 1H NMR(400 MHz, DMSO-d6) d 7.58 (s, IH), 7.54
10 (d, 1H, J = 9.01 Hz), 7.47 (d, 2H, J=7.33 Hz), 7.38 (t, 2H, J= 8.63 Hz), 7.51 (1, 1H, J
= 7.18 Hz), 7.15-7.11 (m, 4H), 7.06 (dd, 2H, J = 9.44 and 2.44 Hz), 5.86 (s, 2H),
5.41 (s, 2H), 5.16 (s, 2H), 2.25 ppm (s, 3H).
Elemental Analysis for C26H21NO4.0.8 H2O;
Calculated: C, 73.33; H, 5.35; N, 3.29.
15 Found: C, 73.16; H. 4.96\ N, 3.20.
6-(Benzyloxy)-1,9dihydropyrano[3.4-b]indole-3,4-dione
20 Step 1
[2-[(Acetyloxy)methyl]-5-(benzyIoxy)-1H-indol-3-yl](oxo)acetic acid
The title compound was prepared from [5-(benzyloxy)-1H-indol-2-yl]methyl acetate
(step 2 of Example 6) and oxalyl chloride following the procedure described in step 5
25 of Example 1. The product was obtained as a brown solid; mp; > 165ºC (dec).
Mass spectrum (ESI. [M-H]-) m/z 366, 1H NMR (400 MHz. DMSO-d6) d 12.42 (s.
1H), 7.51 (s, 1H). 7.48 (d. 2H, J = 7.33 Hz), 7.43 (d. 1H, J = 8.85 Hz), 7.89 (t, 2H. J =
7.33 Hz:). 7.32 (d, 1H, J = 7.17 Hz), 6.99 (d, 1H, J= 7.86 Hz). 5.41 (s, 2H), 5.10 (s.
2H). 2.13 ppm (d, 3H).
30 Elemental Analysis for C20M17NO3.
Calculated: C, 65.39; H. 4.68; N, 3.81.
Found: C, 66.37; H, 4.68; N, 3.83.
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WO 2004/052893 PCT/US2003/038932
Step2
6-(Benzyloxy)-1,9-dihydropyrano[3,4-b]indol-3,4-dione
The title compound was prepared from [2-[(acetyloxy)methyl]-5-(benzyloxy)-1H-indol-
5 3-yl](oxo)acetic acid and aqueous potassium hydroxide in substantially the same
manner, as described in step 6 of Example 1, followed by the treatment with aqueous
HCI, as described in step 7 of Example 1. The product was obtained as a yellow
solid; mp: > 291ºC (dec). Mass spectrum (ESI. [M-H]-) m/z 306. 1H NMR (400
MHZ, DMSO-d6) d 12.37 (s, 1H). 7.54 (s, 1H), 7.49 (s. 1H), 7.43 (d, 2H, J= 8.06 Hz).
10 7.33 (t, 2H, J = 7.64 Hz), 7.32 (d. 1H, J = 7.16 Hz). 7.04 (dd, 1H, J=8.71 and 2.59
Hz), 5.32 (s, 2H). and 5.16 ppm (s, 2H).
Elemental Analysis for C14H13NO4:
Calculated: C, 70.35; H, 4.26: H. 4.56.
Found: C, 70.07; H, 4.17: N, 4.48,
15
Example 7
6-(Benzyloxy)-9-(4-tert-butyIbenzyl)-1,9-dihydropyrano[3,4-b]indole-3,4-dione
Step1
20 [5-(Benzyloxy)-1-(4-tert-butylbenzyl)-1H-indol-2-yl]methyl acetate
The title compound was prepared from [5-(benzyloxy)-1H-indol-2-yI]methyl acetate
and 4-tert-butylbenzyl bromide in substantially the same manner, as described in
step 3 of Example 4. The product was obtained as a white solid; mp: 132-133°C.
25 Mass spectrum (ESI, [M+H]+) m/z 442. 1H NMR (400 MHz, DMSO-d6) d 7.45 (d,
2H, J = 7.33 Hz). 7.36 (1. 2H, J = 7.33 Hz), 7.27-7.27 (m, 4H), 7.16 (d, 1H. J = 2.30
Hz), 6.88-6.84 (m, 3H), 6.53 (s, 1H), 5.36 (a, 2H), 5.18 (g, 2H) 5.06 (s, 2H) 1.77 (s,
3H), and 1.21 ppm(s,9H).
Elemental Analysis for C29H31NO5:
30 Calculated: C, 78.88; H. 7.08; H. 3.17.
Found: C, 78.79; H, 7.07; N. 3.04.
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Step 2
[2-[(AcetyIoxy)methyl]-5-(benzyloxy)-1-(4-tert-butyIbenzyl)-1H-indoI-3-yl](oxo)-
acetic acid
The title compound was prepared from [5-(Benzloxy)-1-(4-tert-butylbenzyl)-1H-indol-
5 2-yl]methyl acetate and oxalyl chloride in substantially the same manner, as
described in step 5 of Example 1. The product was obtained as an off-white solid;
mp: 145-146ºC. Mass spectrum [ESI, [M-H]-] m/z 512. 1H NMR (400 MHz, DMSO-
d6) d 7.61 (d, 1H, J= 2.14 Hz), 7.66 (d, 1H, J = 9.16 Hz),7.48 (d, 2H, J = 7.16 Hz),
7.39 (t, 2H, J = 7.33 Hz) 7.33-7.31 (m, 4H)7.05 (dd, 1H, J= 9.01 and 2.45 Hz), 6.96
10 (d, 1H, J = 8.25 Hz), 5.56 (s, 2H), 6.46 (s, 2H), 5.11 (s, 2H), 1.75 (s, 3H), and 1.21
ppm (s. 9H),
Elemental Analysis for C31H31NO6*0. 7 H2O;
Calculated: C, 70.76; H 6.21; N, 2.66.
Found: C, 70.76; H. 6.39; N, 2.64.
15
Step 3
6-(Benzyloxy)-9-(4-tert-butyIbenzyl)-1,9-dihydropyrano[3,4-b]indole-3,4-dione
The title compound was prepared from [2-{acetyloxy)methyl]-5-(benzyloxy)-1-(4-tert-
20 butylbenzyl)-1H-indol-3-yl](oxo)acetic acid and aqueous potassium hydroxide in
substantially the same manner, as described in step 6 of Example 1, followed by the
treatment with aqueous HCl, as described in step 7 of Example 1. The product was
obtained as an off-white solid; mp: 233-234 ºC. Mass spectrurn (ESI. [M+H]+) m/z
454. 1H NMR (400 MHz, DMSO-d6) d 7.53-7.56 (m, 3H), 7.47 (d, 1H, J= 7.33 Hz),
25 7.39 (t, 2H, J = 7.33 Hz), 7.35-7.30 (m, 3H), 7.15 (d, 2H, J =8.40 Hz), 7.07 (dd, 1H.
J =9.05 and 2.45 Hz), 5.87 (s, 2H), 5.42 (s, 2H). 5.16S (s,2H), and 1.22 ppm (s, 6H)
Elemental Analysis for C22H27NO4:
Calculated: C. 76.30; H. 6.00; N, 3.09.
Found: C, 76.55; H, 6.07; N. 2.98.
30
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WO 2004/052893 PCT/US2003/038932
Example 8
9 -[4-tert-Butylbenzyl)-6-hydroxy-1,9-dihydropyrano[3,4-b]indole-3,4-dione
5 Ethyl 1-(4-tert-butylbenzyl)-5-methoxy-1H-indole-2-carboxylate
The title compound was prepared from ethyl 5-methoxy-1H-indole-2-carboxylate and
4-tert-butyl)benzyl bromide in substantially the same manner, as described in step 1
of Example 1. The product was obtained as a solid. Mass spectrum (ESI, [M-H]-)
10 m/z 364. 1H NMR (400 MHz, DMSO-d6) d 7.48 (d. 1H, J = 9.20 Hz), 7.27 (s, 1H).
7.24 (d, 2H, J = 7.18 Hz), 7.16 (d, 1H, J = 2.44 Hz), 6.96-6.92 (m, 3H) 5.77 (s, 2H)
4.25 (q. 2H), 3.76 (s. 3H), 1.28 (s, 3H), and 1.20 ppm (s, 9H).
Elemental Analysis for C22H27NO3:
Calculated: C, 75.59; H,7.45; N, 3.83.
15 Found: C, 75.04; H, 7.67; N, 3.66.
Step 2
[1-(4-tert-Butylbenzyl)-5-methoxy-1H-indol-2-yl]-6-methanol
20 The title compound was prepared from ethyl 1-(4-tert-butylbenzyl)-6-methoxy-1H-
indole-3-carboxylate and lithium aluminum hydride in substantialIy the same manner,
as described in step 3 of Example 1. The product was obtained as an oil. Mass
spetrum (ESI, [M-H]-) m/z 322. 1H NMR (400 MHz. DMSO-d6) d 7.27 (d. 2H. J =
3.14 Hz), 7.20 (d, 1H, J = 6.70 Hz). 7.01 (6. 1H, J = 2.45 Hz), 6.34 (d, 2H, J = 8.24
25 Hz), 6.65 (dd, 1H, J= 8.05 and 2.44 Hz), 6.34 (s. 1H), 5.37 (s. 2H), 5.26 (t, 1 H), 4.56
(d, ZH, J=4.43 Hz). 3.72 (s. 3H), and 1.21 ppm (s, 9H).
Step 3
[1-(4-tert-Butylbenzyl)-5-methoxy-1H-indol-2-yl]methyl acetate
30
The title compound was prepared from [1-(4-tert-butylbenzyl)-5-methoxy-1H-indol-2-
yl]methanol and acetyl chloride in substantially the same manner, as described in
-34-

WO 2004/052893 PCT/US2003/038932
step 4 of Example 1. The product was obtained as a white solid. Mass spectrum
(ESI, [M+H]+) m/z 366. 1H NMR (400 MHz, DMSO-d6) d 7.30 (s, 1H), 7.28 (d, 2H,
J= 8.10 Hz), 7.07 (d. 1H. J=2.45 Hz), 7.86 (d. 2H, J = 8.24 Hz), 7.76 (dd, 1H. J=
8.86 and 2.29 Hz). 6.54 (s, 1H), 5.36 (s, 2H), 5.18 (s, 2H) 3.74 (s, 3H), 1.77 (s,3H),
5 and 1.20 ppm(s,9H).
Elemental Analysis for C28H27NO5:
Calculated: C, 75.59; H. 7.45; N 3.83.
Found: C. 75.37; H, 7.65; N, 3.70.
10 Step 4
[2{(Acetyloxy)methyl]1-(4-tert-butylbenzyl-5-methoxy-1H-indol-3-yl](oxo)-
acetic acid
The title compound was prepared from [1-(4-tert-butylenzyl)-5-methoxy-1H-indol-2-
15 yl]methyl Acetate and oxalyl chloride in substantially the same manner, as described
to step 5 of Example 1. The product was obtained as a brown solid; mp. > 105ºC
(decomposed). Mass spectrum (ESI, [M-H]-) m/z 436. 1H NMR (400 MHz, DMSO-
d6) d 7.54 (d, 1H, J = 8.10 Hz) 7.51 (d, 1H. J = 1.63 Hz), 7.32 (d,2H,J= 6.25 Hz).
6,97-5.34 (m, 3H), 5.56 (s. 2H), 5.46 (s, 2H). 3.78 (s, 3H), 1.75 (s, 3H), and 1.21 ppm
20 (s. 9H).
Elemental Analysis for C25H27NO6- 0.12 H2O:
Calculated: C. 68.3; H, 6.25; N. 3.19.
Found: C, 69.00; H. 6.66; N, 2.09.
25 Step 5
9-(4-tert-Butylbenzyl)-6-methoxy-[1,9-dihydropyrano[3,4-b]indole-3,4-dione
The title compound was prepared from [2-[(acetyloxy)methyl]-1-(4-tert-butylbenzyl)-5-
methoxy-1H-indol-3-yl](oxo)acetic acid and aqueous, potassium hydroxide in
30 Sustantially the same manner as described in step 6 of Example 1, followed by the
treatment with aq HCl in substantially the same manner as described in step 7 of
Example 1, The product was obtained as an off-white solid, mp: 238-239ºC. Mass
spectrum (ESI, [M-H]-) m/z376. 1H NMR (400 MHz. DMSO-d6) d 7.56 (d, 1H, J =
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WO 2004/052893 PCT/US2003/038932
9.01 HZ) 7.48 (d. 1H. J = 2.45 Hz), 7.35 (d, 2H, J = 8.40 Hz) 7.14 (d. 2H. J = 8.25
Hz), 8.98 (dd, 1H, J = 8.86 and 2.44 Hz) 5.87 (s, 2H), 5.42 (s, 2H), 3.81 (s, 3H), 1.22
ppm(s,9H).
Elemental Analysis for C23H23NO4.
5 Calculated: C, 73.19; H, 6.14: N, 3.71.
Found: C. 73.23:. H, 6.11, H, 3.60.
Step 6
9-{4-tert-Butylbenzyl)-6-hydroxy-1,9-dihydropyrano[3,4-b]indole-3,4-dione
10
Boron tribromide (2.2 mL, 23.3 mmol) was added dropwise to a stirring suspension of
9-(4-tert-Butylbenzyl)-6-methoxy-1,9-dihydropyrano[3,4-b]indole-3,4-dione (4.92 g.
13.0 mmol) in methylene chloride (25 mL) at -78 °C under a nitrogen atmosphere
over a period of 20 minutes. The mixture was then warmed up to room temperature.
15 After stirring at room temperature for 6.5 hours, the reaction mixture was carefully
quenched with a small amount of water and the evaporated to dryness, The rescue
was stirred in either and filtered to afford a brown solid- Crystallization form aqueous''
methanol afforded the title compound as a brown solid (1.72 g). Mass spectrum
(ESI, [M-H]-) m/Z 362. 1HNMR (400 MHz, DMSO-d6): d 9.39 (s, 1H), 7.44 (d, 1H, J =
20 8.85 Hz),7.40 (d, 1H. J = 2.29 Hz),7.34 (d, 2H, J=6.25 Hz),6.80(dd, 1H. J=8.86
and 2.29 Hz). 5.84 (s. 2H), 5.37 (s, 2H), 1.22 ppm (s, 9H).
Elemental Analysis for C22H21NO4* 0.7 H2O:
Calculated; C, 70.27; H, 6.00: N, 3.73.
Found: C, 70.01; H, 5.62; N, 3.81.
25
Example 9
9-Benzyl-6-(4-chlorophenyl)-1,9-dihydropyrano[3,4-b]indole-3,4-dione
Step1
30 (1-Benzyl-5-bromo-1H-indol-2-yl)methanol
The title compound was prepared from ethyl 5-bromo-1-benzyl-1H-indole-2-
carboxylate and lithium aluminum hydride in substantially the same manner, as
described in step 3 of Example 1. The product was obtained as a semi-solid. Mass
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WO 2004/052893 PCT/US2003/038932
spectrum (ESI, [M-H]-) m/z314. 1H NMR (400 MHz, DMSO-d6) d 7.76 (s. 1H),7.58-
7.15 (m, 4H), 7.12 (d, 1H, J =8.69 and 1.97 Hz), 6.98 (d, 2H, J = 8.34 Hz), 6.41 (s,
1H), 5.43 (s, 2H), 5.33 (t, 1H, J = 5.57 Hz), and 4.54 ppm(d, 2H, J=5.43 Hz).
5 Step 2
(1-Benzyl-5-bromo-1H-indol-2-yl)methyl acetate
The title compound was prepared from (1-benzyl-5-bromo-1H-indol-2-yl]methanol
and acatyl chloride in substantially the same manner, as described to step 4 of
10 Example 1. The product was obtained as a semi-solid. Mass spectrum [ESI. [M-H]-)
m/z 356. 1H NMR (400 MHz, DMSO-d6) d 7.78 (d, 1H, J=2-13 Hz), 7.36 (d, 1H, J =
3.86 Hz).7.29-7.20(m, 4H).6.94(d,2H, J= 7.17 Hz),6.54(s. 1H), 5.47 (s.2H),5.21
(s, 2H). and 1.79 ppm (s, 3H).
15 Step 3
[1-Benzyl-5-bromo-2-(hydroxymethyl-1H-indol-3-yl](oxo)acetic acid,
potassium salt
Oxalyl chloride (30.2 mL) was added dropwise to a stirring solution of (4-benzyl-5-
20 bromo-1H-indol-yl)methyl acetate (31.5 g. 88.0 mmol) in THF (890 mL) at room
temperature over a period of 30 minutes under a nitrogen atmosphere. The reaction
was monitored by TLC. More oxayl chloride was added as needed. After the
reaction completed, the reaction mixture was quenched carefully with water and
extracted with ethyl acetate. The organic extracts were washed with water, and
25 brine, dried over anhydrous magnesium sulfate, and evaporated to give a solid. This
solid was treated with aqueous potassium hydroxide (2.0 N, 96.8 mL. 193.6 mmol) in
THF : MeOH (1:1, 800 mL) with stirring at room temperature for 2 h. The mixture
was concentrated to yield a semi-solid. Triluration with water and collection of the
solid by filtration afforded the title compound at a light brown solid (32.2 g), mp:
30 >160ºC (dec); Mass spectrum (ESI, [M-H]-) m/z 366. 1H NMR (400 MHz, DMSO-
d6) d 8.35 (d. 1H, J= 1.99 Hz), 7.40 (d, 1H, J = 8.70 Hz), 7.29-7.26 (m, 4H), 7.23 (d,
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WO 2004/052892 PCT/US2003/038932
2H. J = 7.33Hz).7.10 (d. 2H. J = 7.18 Hz), 5.72 (b, 1H), 5.59 (s, 2H), and 4.81 ppm
(s. 2H).
Elemental Analysis for C20H13BrNO4- 1.0 K 1.0 H20:
Calculated: C, 48.66; H. 3.40; N, 3.15.
5 Found: C, 48.61; H. 3.36; N, 2.89,
Step 4
a-Benzyl-6-(4-chlorophenyl)-1,9-dihydropyrano[3.4-b]indole-3,4-dione.
10 A mixture of [1-benzyl-5-bromo-2(hydromethyl)-1H-indol-3yl](oxo)acetic acid,
potassium salt (3.15 g, 7.4 mmol), 4-chlorophenyIboronic acid (1.74 g, 2.6 mmol),
potassium carbonate (2.55 g, 13.7 mmol), palladium(lI) acetate (0.032 g) and
tetrabutylammonium bromide (2.4 g. 7.4 mmol) in 15 % dioxane in water (45 mL) was
stirred at 70ºC. The reaction was monitored by TLC. After no [1-benzyl-5-bromo-2-
15 (hydroxymethyl)-1H-indol-3-yl](oxo)acetic acid, was detected by TLC, the reaction
was cooled down and solvent was decanted. The dark gum-like oil was partitioned
between ethyl acetate and 10 % aqueous HCl, The upper organic layer was
separated, washed with water, and filtered. This filtrate was evaporated to give a
solid. This solid was triturated with ethyl ether with stirring and dried in vacuum at
20 60°C for 3 hours to afford the titte compound as an off-white solid (0.77 g), m.p. 263-
264ºC. Mass spectrum (ESI. [M+H]+) m/z 402. 1H NMR (400 MHz, DMSO-d6) d
8.19 (s, 1H), 7.71-7.87 (m. 3H), 7.63 (dd, 1H, J = 6.66 and 1.63 Hz). ), 7.49 (dd, 2H.
J = 3.54 and 1.95 Hz). 7.34-7.25 (m, 3H). 7.22 (d, 2H, J = 7.46 Hz), 5.88 (s. 2H), and
5.52 ppm (s. 2H).
25 Elemental Analysis for C24H18ClNO3* 0.4 H2O:
Calculated: C. 70.47; H, 4.14; M, 3.42.
Found: C. 70.57; H, 4.19; N, 3.12,
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WO 2004/052893 PCT/US2003/038932
11 -Benzyl-5-(4-chlorophenyl)-2-[hydroxymethyl)-1H-indol-3-yl](oxo)acetic acid,
potassium salt
5 Step 1
[1-Benzyl-5-(4-chlorophenyl)-2-(hydroxymethyl)-1H-indol-3-yl)(oxo)acetic acid,
potassium salt
A solution of 9-benzyl-6-(4-chlorophenyl)-1,9-hydropyrano[3,4-b]indole-3,4-dione
10 (0.613 g, 0,1.53 mmol) and (1.0 N, 1.6 mL 16 mmol) in THF:MeOH (1:1, 14 mL)
was stirred at room temperature for 2 hours. The reaction was followed by NMR.
More aqueous potassium hydroxide was added as needed. After the starting material
was detected, the reaction mixture was evaporated to dryness. The residual solid
was trituriated with ethyl ether to afford the title compound as a light grey solid, mp:
15 225-226°C; Mass spectrum (ESl, [M-H]-) m/z 416. 1H NMR (400' MHz DMSO-d6) d
8.51 (s, 1H), 7.64 (d, 2H J= 8.55 Hz). 7.52-7.49 (m, 3H), 7.45 (d. 1H, J= 8.55 Hz).
7.31-7.26 (m, 2H), 7.18 (d. 1H, J = 7.64 Hz), 7.15 (d, 2H. J =7.64 Hz), 5.75 (t. 1H,
J = 7.57 Hz)), 5.61 (s, 2H) and 4.81 ppm (d, 2H, J= 5.95 Hz).
Elemental Analysis for C24H17ClNO4* 1.0 K - 1.6 H2O:
20 Calculated: C, 59.22; H, 4.18;N,2.88,
Found: C, 59.02; H, 3.90; N. 2.70.
Example 11
9-Benzyl-6-(3-methylphenyl)-1,9-dihydropyrano[3,4-b]indole-3,4-dione
Step1
The title compound was prepared from [1-benzyl-5-bromo-2-(hydroxymethyl)-1H-
30 indol-3-yl](oxo)acetic acid, potassium salt and m-tolyboronic acid in substantially the
same manner, as described in step 4 of Example 9. The product was obtained as a
grey solid, mp: 325-226ºC; Mass spectrum (ESI. [M+H]+) m/z382 .1H NMR (400
MHz DMSO-d6) d 8.22 (a, 1H), 7.22 (d, 1H, J = 8.40 Hz), 7.65 (dd, 1H. J= 8.45 and
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WO 2004/052893 PCT/US2003/038932

183 Hz), 7.49 (s, 1H). 7.47-7.34 (m, 3H). 7.31 (d. 1H, J=7.33 Hz), 7.27 (d, 2H, J =
8.40 Hz). 7.13 (d, 1H, J = 7.43 Hz), 5.92 (s. 2H), 5.55 (s. 2H), and 2.39 ppm (s, 3H).
Elemental Analysis for C25H19NO3- 0.4 H2O:
Calculated- C, 77.26; H. 5.14 N. 3.60.
5 Found: C, 77.45: H, 5.09; N, 3.51.
Example 12
9-Benzyl-6-(1,1-biphenyl-4-yI)-1,9-dihydropyrano[3,4-b]indole-3,4-dione
10 Step 1
9 -Benzyl-6-(1,1-biphenyl-4-yl)-1,9-dihydropyrane[3,4-b]indole-3,4-dione
The title compound was prepared from [1-benzyl-5-bromo-2-(hydroxymethyl)-1H-
indol-3-yl](oxo)acetic acid, potassium salt and 4-biphenylboronic acid in substantially
15 the same manner, as described in stap 4 of Example 9. The product was obtained
as a white solid, mp: 254-255 ºC; Mass spectrum (ESl, [M+H]+) m/z 444. 1H NMR
(400 MHz, DMSO-d6) d 8.26(s,1H), 7.75-7.68 (m. 8H), 7.45-7.43 (m, 2H), 7.36-7.23
(m,6H) 5.89 (3,2H), 5.53 ppm (3, 2H).
Elemental Analysis for C30H27NO3- 0.2 H2O:
20 Calculated: C, 80.59: H, 4.82; M. 3.13.
Found; C, 80.70,H, 4.60; N, 2.82.
Example 13
[1-Benzyl-5-(1,1-biphenyl-4-yl)-2-(hydroxymethyl)-1H-indol-3-yI](oxo)acetic
25 acid, potassium salt
Step1
[1-Benzyl-5-(1,1'-biphenyl-4-yl)-2-(hydroxymethyl]-1H-indoI-3-yl](oxo)acetic
acid, potassium salt
30
The title compound was prepared from 9-benzyl-6-(1,1*-biphenyl-4-yl)-1,9-
dihydropyrano[3,4-b]indole-3,4-dione and aqueous potassium hydroxide in
substantially the same manner, as described in step 1 of Example 10. The product
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WO 2004/052893 PCT/US2003/038932
was obtained as a light yellow solid, mp: 266-265 ºC; Mass spectrum (ESI, [M-H]-)
m/z 460. 1H NMR (400 MHz, DMSO-d6) d 8.58 (s, 1H), 7.70-7.72 (m, 6H), 7.52 (s,
2H), 7.48 (t. 2H, J = 7.63 Hz), 7.36 (t, 1H, J = 7.33 Hz), 7.32-7.29 (m. 2H), ). 7.24 (t.
1H, J = 7.02 Hz). 7.17 (d, 1H, J = 7.63 Hz), 5.76 (t, 1H), 5.62 (s, 2H). and 4.83 ppm
5 (d, 2H,J=5.34Hz).
Elemental Analysis for C30H23NO4- 1.0 K * 1.50 H2O:
Calculated: C, 68.42; H, 4.79; N, 2.66.
Found: C, 68.24; H, 4.85; N. 2.46.
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WO 2004/052893 PCT/US2003/038932
CLAIMS:
1. A. compound of formulas I and II

5
10 X is hydrogen, an alkali metal or a basic amine moiety
R1 is C1-C6 alkyl, C3-C8 cycloalkyl, -CH2-C3-C6 cycloalkyl, pyridinyl, -CH2-
pyridinyl, phenyl or benyl, wherein the rings of the cycloalkyl, pyridinyl phenyl and
benzol groups may be optionally substituted by forms 1 to 3 groups independently
15 selected from halogen, C1-C5 alkyl, C1-C6 perfluoroalkyl, -O-C1-C5 perfluoroalkyl, C1
C6 alkoxy. -OH. -NH2 or-NO2
R2 is hydrogen, halogen, C1-C6 alkyl, C1-C3 perfluoroalkyl, C1-C5 alkoxy, C1-C6
cycloalkyl. -CH2-C3-C8 cycloalkyl. hydroxy, -NH3, or -NO2:
20
R3 is hydrogen, halogen, C1-C6 alkyl, C1-C3 perfluoroalkyl, C1-C6 alkoxy C3-C6
cycloalkyl, -CH2-C3-C6 cycloalkyl, hydroxy, -NH2. -NO2 phenyl, benzyl, benzyloxy,
pyridinyl, or -CH2-pyridinyl, wherein the rings of these groups may be optionally
substituted by from 1 to 3 groups independently selected from phenyl, halogen, C1-C6
or a pharmaceutically acceptable salt or ester form thereof.
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WO 2004/052893 PCT/US2003/038932

wherein R1, R2 and R3 are as defined in Claim 1 or a pharmaceutical acceptable
5 salt or ester form thereof.
3. A compound according to Claim 1 or 2 wherein R3 is hydroxy, phenyl or
benzyloxy, whereia the rings of the phenyl and benzyloxy groups may be optionally
independently substituted by from 1 to 3 groups independently selected from phenyl,
10 halogen, C1-C3alkyl.C1-C3 perfluoroaIkyl.-O-C1-C3 perfluoroalkyl. C1-C3 alkoxy,-OH,
-NH2 or -NO2; or a pharmaceutically acceptable salt or ester form thereof.
4. A compound according to any one of Claims 1 to 3 wherein R2 is hydrogen or
a pharmaceutically aceeptable salt or ester form thereof.
15
5. A compound according to any one of Claims 1 to 4 wherein R4 is hydrogen or
benzyl, wherein the ring of the benzyl group mat be optionally substituted by from 1
to 3 groups independency selected from halogen, C1-C3 alkyl, C1-C3 perfluoroalkyl,
-O-C1-C3 perfluoroalkyl, C1-C3 alkoxy, -OH, -NH2 or -NO2 or a pharmaceutically
20 acceptable salt or ester form thereof.

25
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WO 2004/052893 PCT/US2003/038932
R, is C1-C6 alkyl, C1-C6 cycloalkyl, -CH2-C3-C6 cycloalkyl, pyridinyl, -CH2-
pyridinyl, phenyl or benzyl wherein the rings of the cycloalkyl, pyridinyl, phenyl and
benzyl groups may be optionally substituted by from 1 to 3 groups independently
selected from halogen, C1-C3 alkyl C1-C6 perfluoroalkyl,-O-C1-C5, perfluoroalkyl. C1-
5 C6 alkoxy, -OH, -NH2 or -NO2;
R2 is hydrogen, halogen, C1-C6 alkyl, C1-C3 perfluoroalkyl. C1-C6 alkoxy, C3-C4
cyclokyl, -CH2-C3-C6 cycloalkyl, NH2 or -NO2,
10 R3 is hydrogen halogen. C1-C5 alkyl, C1-C3 perfluoroalkyl. C1-C5 alkoxy, C3-C6
cycloalkyl, -CH2-C3-C6 cycloalkyl hydroxy, -NH2, -NO2 phenyl, benzyl, benzyloxy,
pyridinyl, or -CH2-pyridinyl, wherein the rings of these groups may be optionally
substituted by from 1 to 3 groups independently selected from halogen. C1-C6 alkyl.
C1-C5 perfluoroalkyl. -O-C1-C6, perfluoroalkyl. C1-C6 alkoxy, -OH, -NH2. or -NO2, or a
15 pharmaceutically acceptable salt or ester form thereof.

20 wherein:
R1 is C1-C6 alkyl C3-C6 cycloalkyl, -CH2-C3-C6 cycloalkyl, pyridinyl, -CH2
pyridinyl, phenyl or benzyl, wherein the rings of the cycloalkyl, pyridinyl, phenyl and
benzyl groups may be optionally substituted by from 1 to 3 groups independently
selected from halogen, C1-C6 alkyl, C1-C6 perfluoroalkyl, -O-C1-C6 perfluoroalkyl- C1-
R2 is hydrogen, halogen. C1-C6 alkyl, C1-C3 perfluoroalkyl, C1-C6 alkoxy. C1-C6
cycloalkyl, -CH2-C3-C6 cycloalkyl, -NH2, or -NO2;
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WO 2004/052893 PCT/US2003/038932
R3 phenyl, benzyl, benzyloxy, pyridinyl, or -CH2-pyridinyl. with the rings or
these groups being optionally substituted by from 1 to 3 groups independently
selected from halogen, C1-C6 alkyl, C1-C6 perfluoroalkyl, -O-C1-C6 pefluoroalkyl, C1-
C6 alkoxy, -OH, -NH2 or -NO2; or a pharmaceutically acceptable salt or ester form
5 thereof.

10 wherein:
R1 is C1-C6 alkyl preferably C1-C6 alkyl, C1-C6 cycloalkyl, -CH2-C3-C6
cycloalkyl, or benzyl, wherein the rings of the cycloalkyl and benzyl groups may be
optionally substituted by from 1 to 3 groups selected independently from halogen, C1-
C6 alkyl, C1-C6 perfluorocalkyl -O-C1-C6 perfluoroalkyl, C1-C6 alkoxy, -OH. -NH2, or
R2 is hydrogen, halogen, C1-C3 alkyl, C1-C6 perfluoroalkyl, preferably -CF3
C1-C6 alkoxy, C3-C6 cycloalkyl, -CH2-C3-C6 cycloalkyl, hydroxy, -NH2 or-NO2;
20 R4 R5 and R6 are each independently hydrogen, phenyl, halogen, C1-C3 alkyl,
C1-C3 perfluoroalkyl, -O-C1-C3 perfluoroalkyl, C1-C3 alkoxy. -OH. -NH2 or-NO2; or a
pharmaceutically acceptable salt or ester form thereof.
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WO 2004/052893 PCT/US2003/038932

Wherein:
5 R1 is C1-C3 alkyl, C3-C6 cycloalkyl, -CH2-C3-C6 cycloalkyl, or benzyl wherein
the rings of the cycloalkyl and benzyl groups may be optionally substituted by from 1
to 3 groups independency selected from halogen. C1-C3 alkyl, C1-C6 perfluoroalkyl,
preferably -CF3 -O-C1-C3 perfluoroalkyl, preferably -O-CF3, C1-C6 alkoxy, -OH,
-NH2 or -NO2;
10
R2 is hydrogen, halogen C1-C3 aIkyI C1-C3 perfluoroalkyl, preferably -CF3,
C1-C3 alkoxy, C3-C6 cycloalkyl -CH2-C3-C6 cycloalkyl, hydroxy, -NH2 or -NO2; and
R4, R5 and R6 are each independently hydrogen, phenyl halogen, C1-C3 alkyl,
15 C1-C3 perfluoroalkyl, preferably -CF3 -O-C1-C3 perfluoroalkyl, preferably -O-CF3 C1
C3 alkoxy. -OH, -NH2 or -NO2 or a pharmaceutically acceptable salt or ester form
10. The compound of Claim 1 which is any one of
20 9-(4-Methylbenzyl)-6-[4(trifluoromethoxy)phenyl]-1,9-dihydropyrano[3,4-b]indole-3,4-
dione.
9-Benzyl-6-[4-(trifluoromethoxy)phenyl]-1,9-dihydropyrano[3,4-b]indole-3,4-dione,
9-(4-Methylbenzyl)-6-(3-Methylphenyl)-1,9-dyhydropyrano[3,4-b]indole-3,3-dione,
9-(4-tert-butybenzyl)-6-(3-Methylphenyl)-1,9-dihydropyrano[3,4-b]indole-3,4-dione,
6-(Benzyloxy)-1,9-dihydropyrano[3,4,-b]indole-3,4-dione,
6-(Benzyloxy)-9-(4-tertbutylbenzyl)-1,9-dihydropyrano[3,4-b]indole-3,4-dione,
9-(4-tertbutybenzyl)-6-hydroxy-1,9-dihydropyrano[3,4-b]indole-3,4-dione,
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WO 2004/052893 PCT/US2003/038932
9-benzyl-6-(4-chlorophenyl)-1,9-dihydropyrano[3,4-b]indole-3,4-dione,
[1-benzyl-5-(4-chlorophenyl)-2-(hydroxymethyl)-1H-indole-2-yl](oxo)acetic acid
9-benzyl-6-(3-methylphenyl)-1,9-dihydropyrano[3,4-b]indole-3,4-dione,
9-benzyl-6-(1-1-bi-phenyl-4-yl)-1,9-dihydropyrano[3,4-b]indole-3,4-dione, or
5 [1-benzyl-5-(1,1-diphenyl-1-yl)-2-(hydroxymethyl)-1H-indole-3-yl] (oxo)acetic acid,
or a pharmaceutically acceptable salt or ester form thereof.
11. A method of inhibiting plasminogen activator inhibitor in a mammal
comprising administering to a mammal in need thereof a therapeutically effective
10 amount of a compound as claimed in any one of Claims 1 to 10
12. A pharmaceutical composition comprising a compound as claimed in
anyone of Claims 1 to 10 and a pharmaceutical carrier
15 13. A method for treatment of thrombosis or fibrinolytic impairment in a
mammal, the method comprising administering to a mammal in need thereof a
pharmaceutically effective ammount of a compound as claimed in any one of Claims 1
20 14. A method of Claim 13 wherein the thrombosis or fibrinolytic
impairment is associated with formation of atherosclerotic plaques, venous and
arterial thrombosis, myocardial ischemia, atrial fibrination, deep vein thrombosis,
coagulation syndromes, pulmonary fibrosis, cerebral thrombosis, thromboembolic
Complications of surgery or peripheral arterial occlusion.
25
15. A method for the treatment of peripheral arterial disease in a mammal,
cormprising admistering to a mammal in need thereof a pharmaceutically effective
amount of a compound as claimed in any one of Claims 1 to 10,
30 16. A method for the treatment of stroke associated with or resulting from
atrial fibrillation in a mammal, comprising administering to a mammal in need thereof
a pharmaceutically effective amount of a compound as claimed in any one of Claims
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WO 2004/052893 PCT/US2003/038932
17. A method for the treatment of deep vein thrombosis in a mammal.
comprising administering to a mammal in need thereof a pharmaceutical effective
amount of 3 compound as claimed in any one of Claims 1 to 10.
5
18. A method for the treatment of myocardial ischemia in a mammal,
comprising administering to a mammal in need thereof a pharmaceutieally effective
amount of a cmpound as claimed in any one of Claims 1 to 10.
10 19. A method for the treatment of a cardiovascular disease caused by
noninsulin dependent diabetas melitus in a mammal, comprising administering to a
mammal in need thereof a pharmaceutically effective amount of a compound of
Claim 1.
15 20. A method for the treatment of the formation of atherosclerotic plaques
in a mammal, comprising administering to a mammal in need thereof a
pharmaceutically effective amount of a compound as claimed in any one of Claims 1
to 10.
20 21. A method for the treatment of chronic obstructive pulmonary disease
in a mammal, comprising administering to a mamma in need thereof a
pharmaceuitically effective amount of a comnoand as claimed in any one of Claims 1
25 22. A method for the treatment of renal fibrosis in a mammmal, comprising
administering to a mammal in need thereof a pharmaceutically effective amount of a
compound as claimed in any one of Claims 1 to 10.
23. A method for the treatment of polycystic ovary syndrome in a
30 mammal, comprising administering to a mammal in need thereof a pharmaceutically
effective amount of a compound as claimed in any one of claims 1 to 10.
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WO 2004/052893 PCT/US2003/038932
24. A method for the treatment of Alzheimer's disease in a mammal.
comprising administering to a mammal in need thereof a pharmaceutically effective
amount of a compound as claimed in any one of 1 to 10.
5 25. A method for the treatment of cancer in a mammal, comprising
administering to a mammal in need thereof a pharmaceutically effective amount of a
compound as claimed in any one of Claim 1 to 10.

10
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Compound of formula (I) and (II) are provided wherein X is an alkali metel or a basic amine moitey; R1is alkyl
cycloalkyl, -CH2-cycloalkyl, pyridinyl, phenyl or benzyl, the rings of these groops being optionally substituted; R2
is H, halogen, alkyl, perfluoroalkyl, alkoxy, cycloalkyl, -CH2,cycloalkyl, -NH2- or -NO2; R3 is phenyl, benzyl, benzyloxy,pyridinyl,
or -CH2-pyridinyl, with the rings of these groups being optionally substituted; or a pharmaceutically acceptable with or ester form
thereof, as well as pharmaceutically composition and methods using these compounds as inhibitors of plasmogen activator in-
hibitor-1 (PAI-1) and as therapeutic compositions for heating conditions resulting from fibrinolytic disorders such as deep vein
thrombosis and coronary heart disease and pulmonary fibrosis.