Patent Application
20070299125
The present invention relates generally to substituted indoles and methods of using them.
The serine protease inhibitor PAI-1 is one of the primary inhibitors of the fibrinolytic system. The fibrinolytic system includes the proenzyme plasminogen, which is converted to the active enzyme, plasmin, by one of two tissue type plasminogen activators, t-PA or u-PA. PAI-1 is the principal physiological inhibitor of t-PA and u-PA. One of plasmin's main responsibilities in the fibrinolytic system is to digest fibrin at the site of vascular injury. The fibrinolytic system, however, is not only responsible for the removal of fibrin from circulation but is also involved in several other biological processes including ovulation, embryogenesis, intima proliferation, angiogenesis, tumorigenesis, and atherosclerosis.
Elevated levels of PAI-1 have been associated with a variety of diseases and conditions including those associated with impairment of the fibrinolytic system. For example, elevated levels of PAI-1 have been implicated in thrombotic diseases, e.g., diseases characterized by formation of a thrombus that obstructs vascular blood flow locally or detaches and embolizes to occlude blood flow downstream. (Krishnamurti, Blood, 69, 798 (1987); Reilly, Arteriosclerosis and Thrombosis, 11, 1276 (1991); Carmeliet, Journal of Clinical Investigation, 92, 2756 (1993), Rocha, Fibrinolysis, 8, 294, 1994; Aznar, Haemostasis 24, 243 (1994)). Antibody neutralization of PAI-1 activity resulted in promotion of endogenous thrombolysis and reperfusion (Biemond, Circulation, 91, 1175 (1995); Levi, Circulation 85, 305, (1992)). Elevated levels of PAI-1 have also been implicated in diseases such as polycystic ovary syndrome (Nordt, Journal of clinical Endocrinology and Metabolism, 85, 4, 1563 (2000)), bone loss induced by estrogen deficiency (Daci, Journal of Bone and Mineral Research, 15, 8, 1510 (2000)), cystic fibrosis, diabetes, chronic periodontitis, lymphomas, diseases associated with extracellular matrix accumulation, malignancies and diseases associated with neoangiogenesis, inflammatory diseases, vascular damage associated with infections, and diseases associated with increased uPA levels such as breast and ovarian cancer.
In view of the foregoing, there exists a need for the identification of inhibitors of PAI-1 activity and for methods of using the identified inhibitors to modulate PAI-1 expression or activity in a subject in order to treat disorders associated with elevated PAI-1 levels.
The present invention provides substituted indoles and methods of using them. In certain embodiments, substituted 1-aralkyl or 1-aryl-1H-indoles are provided including those of the following formula:
wherein:
X is a bond or C1-C6 alkylene;
R1 and R2 are, independently, hydrogen, C1-C6 alkyl, C2-C7 alkenyl, C2-C7 alkynyl, C3-C8 cycloalkyl, C7-C11 bicycloalkyl, C6-C10 aryl, heterocycle, —C(═O)C1-C6 alkyl, —C(═O)aryl, —C(═O)heterocycle, —C(═O)N(R6)C1-C6 alkyl, —C(═O)N(R6)aryl, —C(═O)N(R6)heterocycle, —SO2—C1-C6 alkyl, —SO2-aryl, or —SO2-heterocycle;
or R1 and R2 together form a heterocycle;
R3 and R4 are independently, hydrogen, halogen, C1-C6 alkyl, C1-C3 perfluoroalkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, —C(═O)C1-C3 alkyl, —OH, —NH2, or —NO2;
R5 is hydrogen, C1-C6 alkyl, C2-C7 alkenyl, C2-C7 alkynyl, C3-C8 cycloalkyl, C7-C11 bicycloalkyl, C6-C10 aryl, or heterocyle;
A is hydrogen, C6-C10 aryl, or heterocyle; and
R6 is hydrogen, C1-C6 alkyl, halogen, C2-C7 alkenyl, C2-C7 alkynyl, C3-C8 cycloalkyl, aralkyl, C6-C10 aryl, heterocycle, hydroxy, C1-C6 alkoxy, aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO2H, —OCO2C1-C6 alkyl, —CO2C1-C6 alkyl, —CO2-aryl, —CO2(C1-C6 alkyl)aryl, —OCO2-aryl, —C(═O)NH2, —C(═O)NHOH, amino, C1-C6 alkylamino, dialkylamino of 1-6 carbons per alkyl moiety, —NHC(═O)NH—C1-C6 alkyl, —NHSO2—C1-C6 alkyl, —NHSO2-aryl, or —NHSO2-heterocycle.
The present invention also provides, inter alia, pharmaceutically acceptable salt or ester forms of Formula 1.
The present invention further provides, inter alia, methods of using substituted indoles. In one aspect of the present invention, a therapeutically effective amount of one or more substituted indoles is administered to a subject in order to treat a PAI-1 related disorder, e.g., by inhibiting PAI-1 activity in the subject. PAI-1 activity is associated with a number of diseases and conditions. For example, in one embodiment of the present invention, PAI-1 activity is associated with impairment of the fibrinolytic system. In other embodiments, PAI-1 activity is associated with thrombosis, e.g., venous thrombosis, arterial thrombosis, cerebral thrombosis, and deep vein thrombosis, atrial fibrillation, pulmonary fibrosis, thromboembolic complications of surgery, cardiovascular disease, e.g., myocardial ischemia, atherosclerotic plaque formation, chronic obstructive pulmonary disease, renal fibrosis, polycystic ovary syndrome, Alzheimer's disease, or cancer.
A. General Overview
The present invention provides compounds that inhibit PAI-1 activity, processes for preparing such compounds, pharmaceutical compositions containing such compounds, and methods for using such compounds in medical therapies. The compounds have properties that are useful for the treatment, including the prevention and inhibition, of a wide variety of diseases and disorders including those involving the production and/or action of PAI-1. These include disorders resulting from impairment of the fibrinolytic system including, but not limited to, thrombosis, coronary heart disease, renal fibrosis, atherosclerotic plaque formation, pulmonary disease, myocardial ischemia, atrial fibrillation, coagulation syndromes, thromboembolic complications of surgery, peripheral arterial occlusion and pulmonary fibrosis. Other disorders include, but are not limited to, polycystic ovary syndrome, Alzheimer's disease, and cancer.
The terms “alkyl” and “alkylene,” as used herein, whether used alone or as part of another group, refer to substituted or unsubstituted aliphatic hydrocarbon chains, the difference being that alkyl groups are monovalent (i.e., terminal) in nature whereas alkylene groups are divalent and typically serve as linkers. Both include, but are not limited to, straight and branched chains containing from 1 to about 12 carbon atoms, preferably 1 to about 6 carbon atoms, unless explicitly specified otherwise. For example, methyl, ethyl, propyl, isopropyl, butyl, i-butyl and t-butyl are encompassed by the term “alkyl.” Specifically included within the definition of “alkyl” are those aliphatic hydrocarbon chains that are optionally substituted. In representative embodiments of the present invention, optional substituents can include C1-C6 alkyl, halogen, C2-C7 alkenyl, C2-C7 alkynyl, C3-C8 cycloalkyl, aralkyl, aryl optionally substituted with R7, heterocycle optionally substituted with R7, hydroxy, C1-C6 alkoxy, aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO2H, —OCO2C1-C6 alkyl, —CO2C1-C6 alkyl, —CO2-aryl, —CO2(C1-C6 alkyl)aryl, —OCO2-aryl, —C(═O)NH2, —C(═O)NHOH, amino, alkylamino, dialkylamino, —NHC(═O)NH—C1-C6 alkyl, —NHSO2—C1-C6 alkyl, —NHSO2-aryl, and —NHSO2-heterocycle.
R7 is halogen, C1-C6 alkoxy, C1-C6 alkyl, C2-C7 alkenyl, C2-C7 alkynyl, hydroxy, —C(═O)C1-C7 alkyl, —SO2—C1-C6 alkyl, —CO2—C1-C6 alkyl, or alkoxycarbonylalkyl. In some embodiments, R7 includes C2-7 acyl.
The carbon number as used in the definitions herein refers to carbon backbone and carbon branching, but does not include carbon atoms of the substituents, such as alkoxy substitutions and the like.
The term “alkenyl”, as used herein, whether used alone or as part of another group, refers to a substituted or unsubstituted aliphatic hydrocarbon chain and includes, but is not limited to, straight and branched chains having 2 to about 10 carbon atoms (unless explicitly specified otherwise) and containing at least one double bond. Preferably, the alkenyl moiety has 1 or 2 double bonds. Such alkenyl moieties can exist in the E or Z conformations and the compounds of this invention include both conformations. Specifically included within the definition of “alkenyl” are those aliphatic hydrocarbon chains that are optionally substituted. In representative embodiments of the present invention, optional substituents can include C1-C6 alkyl, halogen, C2-C7 alkenyl, C2-C7 alkynyl, C3-C8 cycloalkyl, aralkyl, aryl optionally substituted with R7, heterocycle optionally substituted with R7, hydroxy, C1-C6 alkoxy, aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO2H, —OCO2C1-C6 alkyl, —CO2C1-C6 alkyl, —CO2-aryl, —CO2(C1-C6 alkyl)aryl, —OCO2-aryl, —C(═O)NH2, —C(═O)NHOH, amino, alkylamino, dialkylamino, —NHC(═O)NH—C1-C6 alkyl, —NHSO2—C1-C6 alkyl, —NHSO2-aryl, and —NHSO2-heterocycle. Heteroatoms, such as O or S attached to an alkenyl should not be attached to a carbon atom that is bonded to a double bond.
The term “acyl”, employed alone or in combination with other terms, is defined herein as, unless otherwise stated, either an arylalkyl, heteroarylalkyl, (C2-C10) straight chain, or (C4-C11) branched-chain monovalent hydrocarbon moiety; wherein the carbon atom, covalently linked to the defined chemical structure, is oxidized to the carbonyl oxidation state. Such hydrocarbon moieties may be mono or polyunsaturated, and may exist in the E or Z configurations. The compounds of this invention are meant to include all possible E and Z configurations. Examples of acyl moieties include, but are not limited to, chemical groups such as acetyl, propionyl, butyryl, 3,3-dimethylbutyryl, trifluoroacetyl, pivaloyl, hexanoyl, hexenoyl, decanoyl, benzoyl, nicotinyl, isonicotinyl, and homologs, isomers, and the like
The term “alkynyl”, as used herein, whether used alone or as part of another group, refers to a substituted or unsubstituted aliphatic hydrocarbon chain and includes, but is not limited to, straight and branched chains having 2 to about 10 carbon atoms (unless explicitly specified otherwise) and containing at least one triple bond. Preferably, the alkynyl moiety has about 2 to about 7 carbon atoms. In certain embodiments, the alkynyl can contain more than one triple bond and, in such cases, the alknyl group must contain at least three carbon atoms. In representative embodiments of the present invention, optional substituents can include C1-C6 alkyl, halogen, C2-C7 alkenyl, C2-C7 alkynyl, C3-C8 cycloalkyl, aralkyl, aryl optionally substituted with R7, heterocycle optionally substituted with R7, hydroxy, C1-C6 alkoxy, aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO2H, —OCO2C1-C6 alkyl, —CO2C1-C6 alkyl, —CO2-aryl, —CO2(C1-C6 alkyl)aryl, —OCO2-aryl, —C(═O)NH2, —C(═O)NHOH, amino, alkylamino, dialkylamino, —NHC(═O)NH—C1-C6 alkyl, —NHSO2—C1-C6 alkyl, —NHSO2-aryl, and —NHSO2-heterocycle. Heteroatoms, such as O or S attached to an alkynyl should not be attached to the carbon that is bonded to a triple bond.
The term “cycloalkyl” as used herein, whether alone or as part of another group, refers to a substituted or unsubstituted alicyclic hydrocarbon group having 3 to about 20 carbon atoms (unless explicitly specified otherwise), preferably 3 to about 8 carbon atoms. Specifically included within the definition of “cycloalkyl” are those alicyclic hydrocarbon groups that are optionally substituted. In representative embodiments of the present invention, optional substituents can include C1-C6 alkyl, halogen, C2-C7 alkenyl, C2-C7 alkynyl, C3-C8 cycloalkyl, aralkyl, aryl optionally substituted with R7, heterocycle optionally substituted with R7, hydroxy, C1-C6 alkoxy, aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO2H, —OCO2C1-C6 alkyl, —CO2C1-C6 alkyl, —CO2-aryl, —CO2(C1-C6 alkyl)aryl, —OCO2-aryl, —C(═O)NH2, —C(═O)NHOH, amino, alkylamino, dialkylamino, —NHC(═O)NH—C1-C6 alkyl, —NHSO2—C1-C6 alkyl, —NHSO2-aryl, and —NHSO2-heterocycle.
The term “aryl”, as used herein, whether used alone or as part of another group, is defined as a substituted or unsubstituted aromatic hydrocarbon ring group having 5 to about 50 carbon atoms (unless explicitly specified otherwise) with from about 6 to about 10 atoms being preferred. The “aryl” group can have a single ring or multiple condensed rings. The term “aryl” includes, but is not limited to phenyl, α-naphthyl, β-naphthyl, biphenyl, anthryl, tetrahydronaphthyl, fluorenyl, indanyl, biphenylenyl, and acenaphthenyl. Specifically included within the definition of “aryl” are those aromatic groups that are optionally substituted. For example, in representative embodiments of the present invention, the, “aryl” groups are optionally substituted with from 1 to 5 substituents selected from the group consisting of acyloxy, hydroxy, aryl, acyl, alkanoyl, 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 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 thioalkoxy of from 1 to 6 carbon atoms, and trihalomethyl. In an exemplary embodiment of the present invention, the optional substituents include C1-C6 alkyl, halogen, C2-C7 alkenyl, C2-C7 alkynyl, C3-C8 cycloalkyl, aralkyl, aryl optionally substituted with R7, heterocycle optionally substituted with R7, hydroxy, C1-C6 alkoxy, C1-C6 perfluoroalkoxy, aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO2H, —OCO2C1-C6 alkyl, —CO2C1-C6 alkyl, —CO2-aryl, —CO2(C1-C6 alkyl)aryl, —OCO2-aryl, —C(═O)NH2, —C(═O)NHOH, amino, alkylamino, dialkylamino, —NHC(═O)NH—C1-C6 alkyl, —NHSO2—C1-C6 alkyl, —NHSO2-aryl, and —NHSO2-heterocycle.
The term “alkoxy” as used herein, refers to the group Ra—O— wherein Ra is an alkyl group as defined above. Specifically included within the definition of “alkoxy” are those alkoxy groups that are optionally substituted.
The term “aryl-oxy” as used herein, refers to the group Rb—O— wherein Rb is an aryl group as defined above.
The term “alkanoyl” as used herein, refers to the group —C(═O)-alkyl group wherein alkyl is defined as above. Exemplary alkanoyl groups include, but are not limited to, acetyl(ethanoyl), n-propanoyl, n-butanoyl, 2-methylpropanoyl, n-pentanoyl, 2-methylbutanoyl, 3-methylbutanoyl, 2,2-dimethylpropanoyl, heptanoyl, and decanoyl. The alkyl moieties of alkanoyl groups can be optionally substituted.
The term “arylalkyl” or “aralkyl” refers to the group —Ra—Rb, where Ra is an alkyl group as defined above, substituted by Rb, an aryl group, as defined above. Preferably the alkyl group has from 1 to 6 carbon atoms. Examples of arylalkyl moieties include, but are not limited to, benzyl, 1-phenylethyl, 2-phenylethyl, 3-phenylpropyl, 2-phenylpropyl and the like.
The term “alkoxycarbonylalkyl”, as used herein, refers to the group Rc—O—C(═O)Rc—, wherein Rc is an alkyl group as defined above. Preferably, the alkoxycarbonylalkyl has from 3 to 13 carbon atoms.
The term “alkylamino”, as used herein, refers to the group Rc—NH—, wherein Rc is an alkyl group as defined above preferably having from 1 to 6 carbon atoms.
The term “dialkylamino” as used herein refers to the group —N(C1-C6 alkyl)2.
The term “—SO2—C1-C6 alkyl”, as used herein, refers to the group —S(O2)—Ra, wherein Ra is an alkyl group of 1 to 6 carbons as defined above.
The term “bicycloalkyl” refers to an optionally substituted, alkyl group having two bridged rings in its structure and having from about 7 to about 20 carbon atoms (unless explicitly specified otherwise) (and all combinations and subcombinations of ranges and specific numbers of carbon atoms therein), with from about 7 to about 11 carbon atoms being preferred. Exemplary bicycloalkyl-ring structures include, but are not limited to, norbornyl, bornyl, [2.2.2]-bicyclooctyl, cis-pinanyl, trans-pinanyl, camphanyl, iso-bornyl, and fenchyl. Representative substituents include, for example, C1-C6 alkyl, halogen, C2-C7 alkenyl, C2-C7 alkynyl, C3-C8 cycloalkyl, aralkyl, aryl optionally substituted with R7, heterocycle optionally substituted with R7, hydroxy, C1-C6 alkoxy, aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO2H, —OCO2C1-C6 alkyl, —CO2C1-C6 alkyl, —CO2-aryl, —CO2(C1-C6 alkyl)aryl, —OCO2-aryl, —C(═O)NH2, —C(═O)NHOH, amino, alkylamino, dialkylamino, —NHC(═O)NH—C1-C6 alkyl, —NHSO2—C1-C6 alkyl, —NHSO2-aryl, and —NHSO2-heterocycle.
The term “heterocycle”, as used herein, whether used alone or as part of another group, refers to a stable 3 to about 10-member ring containing carbons atoms and from 1 to 4 heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur. A heterocycle of this invention can be either a monocyclic or bicyclic ring system, and can be either saturated, unsaturated, or partially saturated. A heterocycle can be optionally fused to a phenyl ring. Heterocycle groups include, but are not limited to, aziridinyl, azetidinyl, 1,4-dioxanyl, hexahydroazepinyl, piperazinyl, piperidinyl, pyrrolidinyl, morpholinyl, thiomorpholinyl, dihydrobenzimidazolyl, dihydrobenzofuranyl, dihydrobenzothienyl, dihydrobenzoxazolyl, dihydrofuranyl, dihydroimidazolyl, dihydroindolyl, dihydroisooxazolyl, dihydroisothiazolyl, dihydrooxadiazolyl, dihydrooxazolyl, dihydropyrrazinyl, dihydropyrazolyl, dihydropyridinyl, dihydropyrimidinyl, dihydropyrrolyl, dihydroquinolinyl, dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl, dihydrothienyl, dihydrotriazolyl, dihydroazetidinyl, dihydro-1,4-dioxanyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydroquinolinyl, and tetrahydroisoquinolinyl. Preferred heterocycle moieties include: (a) 6-membered saturated, partially unsaturated, or unsaturated heterocycles containing 1-2 nitrogens, optionally fused to a phenyl ring; (b) 5-membered saturated, partially saturated, or unsaturated heterocycles containing 1-3 nitrogen, oxygen, or sulfur atoms, optionally fused to a phenyl ring; (c) saturated, partially unsaturated, or unsaturated bicyclic heterocycles containing 1-4 nitrogen, oxygen, or sulfur atoms; (d) carbazole, dibenzofuran, and dibenzothiophene. Specifically included in the definition of “heterocycle” are those heterocycles that are optionally substituted. Representative substituents include, for example, C1-C6 alkyl, halogen, C2-C7 alkenyl, C2-C7 alkynyl, C3-C8 cycloalkyl, aralkyl, aryl optionally substituted with R7, heterocycle optionally substituted with R7, hydroxy, C1-C6 alkoxy, aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO2H, —OCO2C1-C6 alkyl, —CO2C1-C6 alkyl, —CO2-aryl, —CO2(C1-C6 alkyl)aryl, —OCO2-aryl, —C(═O)NH2, —C(═O)NHOH, amino, alkylamino, dialkylamino, —NHC(═O)NH—C1-C6 alkyl, —NHSO2—C1-C6 alkyl, —NHSO2-aryl, and —NHSO2-heterocycle.
The term “heteroaryl” as used herein is defined as a substituted or unsubstituted aromatic heterocyclic ring system (monocyclic or bicyclic). Heteroaryl groups can have, for example, from about 3 to about 50 carbon atoms (unless explicitly specified otherwise) with from about 4 to about 10 being preferred. In some embodiments, heteroaryl groups are aromatic heterocyclic rings systems having about 4 to about 14 ring atoms including carbon atoms and 1, 2, 3, or 4 heteroatoms selected from oxygen, nitrogen or sulfur. Representative heteroaryl groups are furan, thiophene, indole, azaindole, oxazole, thiazole, isoxazole, isothiazole, imidazole, N-methylimidazole, pyridine, pyrimidine, pyrazine, pyrrole, N-methylpyrrole, pyrazole, N-methylpyrazole, 1,3,4-oxadiazole, 1,2,4-triazole, 1-methyl-1,2,4-triazole, 1H-tetrazole, 1-methyltetrazole, benzoxazole, benzothiazole, benzofuran, benzisoxazole, benzimidazole, N-methylbenzimidazole, azabenzimidazole, indazole, quinazoline, quinoline, and isoquinoline. Bicyclic aromatic heteroaryl groups include phenyl, pyridine, pyrimidine or pyridizine rings that are (a) fused to a 6-membered aromatic (unsaturated) heterocyclic ring having one nitrogen atom; (b) fused to a 5- or 6-membered aromatic (unsaturated) heterocyclic ring having two nitrogen atoms; (c) fused to a 5-membered aromatic (unsaturated) heterocyclic ring having one nitrogen atom together with either one oxygen or one sulfur atom; or (d) fused to a 5-membered aromatic (unsaturated) heterocyclic ring having one heteroatom selected from O, N or S. Specifically included within the definition of “heteroaryl” are those aromatic groups that are optionally substituted. Accordingly, the heteroaryl groups (e.g., pyridinyl) described herein refer to both unsubstituted or substituted groups. In representative embodiments of the present invention, the, “heteroaryl” groups are optionally substituted with 1 to 5 substituents selected from the group consisting 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 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 thioalkoxy of from 1 to 6 carbon atoms, and trihalomethyl.
The term “perfluoroalkyl”, as used herein, whether used alone or as part of another group, refers to a saturated aliphatic hydrocarbon having 1 to 6 carbon atoms and two or more fluorine atoms and includes, but is not limited to, straight or branched chains, such as —CF3, —CH2CF3, —CF2CF3 and —CH(CF3)2.
The term “halogen” or “halo” refers to chlorine, bromine, fluorine, and iodine.
The term “treating” or “treatment” refers to any indicia of success in amelioration of an injury, pathology, or condition, including any objective or subjective parameter such as abatement; remission; diminishing of symptoms or making the injury, pathology, or condition more tolerable to the patient; slowing in the rate of degeneration or decline; making the final point of degeneration less debilitating; or improving a subject's physical or mental well-being. The treatment or amelioration of symptoms can be based on objective or subjective parameters; including the results of a physical examination, neurological examination, and/or psychiatric evaluation. “Treating” or “treatment of a PAI-1 related disorder” includes preventing the onset of symptoms in a subject that may be predisposed to a PAI-1 related disorder but does not yet experience or exhibit symptoms of the disorder (prophylactic treatment), inhibiting the symptoms of the disorder (slowing or arresting its development), providing relief from the symptoms or side-effects of the disorder (including palliative treatment), and/or relieving the symptoms of the disorder (causing regression). Accordingly, the term “treating” includes the administration of the compounds or agents of the present invention to a subject to prevent or delay, to alleviate, or to arrest or inhibit development of the symptoms or conditions associated with PAI-1 related disorders, e.g., tumor growth associated with cancer. A skilled medical practitioner will know how to use standard methods to determine whether a patient is suffering from a disease associated with enhanced levels and/or activity of PAI-1, e.g., by examining the patient and determining whether the patient is suffering from a disease known to be associated with elevated PAI-1 levels or activity or by assaying for PAI-1 levels in blood plasma or tissue of the individual suspected of suffering from a PAI-1 related disease and comparing PAI-1 levels in the blood plasma or tissue of the individual suspected of suffering from a PAI-1 related disease to PAI-1 levels in the blood plasma or tissue of a healthy individual. Increased PAI-1 levels are indicative of disease. Accordingly, the present invention provides, inter alia, methods of administering a compound of the present invention to a subject and determining levels of PAI-1 in the subject. The level of PAI-1 in the subject can be determined before and/or after administration of the compound.
In healthy individuals, PAI-1 is found at low levels in the plasma (from about 5-26 ng/mL), but it is elevated in many PAI-1 related disorders, including, for example, atherosclerosis (Schneiderman J. et. al, Proc Natl Acad Sci 89: 6998-7002, 1992) deep vein thrombosis (Juhan-Vague I, et. al, Thromb Haemost 57: 67-72, 1987), and non-insulin dependent diabetes mellitus (Juhan-Vague I, et. al, Thromb Haemost 78: 565-660, 1997). PAI-1 stabilizes both arterial and venous thrombi, contributing respectively to coronary arterial occlusion in post-myocardial infarction (Hamsten A, et. al. Lancet 2:3-9, 1987), and venous thrombosis following post-operative recovery from orthopedic surgery. (Siemens H J, et. al, J Clin Anesthesia 11: 622-629, 1999). Plasma PAI-1 is also elevated, for example, in postmenopausal women, and has been proposed to contribute to the increased incidence of cardiovascular disease in this population (Koh K et. al, N Engl J Med 336: 683-690, 1997).
The term “PAI-1 related disorder or disease” refers to any disease or condition that is associated with increased or enhanced expression or activity of PAI-1 or increased or enhanced expression or activity of a gene encoding PAI-1. Examples of such increased activity or expression can include one or more of the following: activity of the protein or expression of the gene encoding the protein is increased above the level of that in normal subjects; activity of the protein or expression of the gene encoding the protein is in an organ, tissue or cell where it is not normally detected in normal subjects (i.e. spatial distribution of the protein or expression of the gene encoding the protein is altered); activity of the protein or expression of the gene encoding the protein is increased when activity of the protein or expression of the gene encoding the protein is present in an organ, tissue or cell for a longer period than in a normal subjects (i.e., duration of activity of the protein or expression of the gene encoding the protein is increased). A normal or healthy subject is a subject not suffering from a PAI-1 related disorder or disease.
The term “pharmaceutically acceptable excipient” means an excipient that is useful in preparing a pharmaceutical composition that is generally safe, non-toxic, and desirable, and includes excipients that are acceptable for veterinary use as well as for human pharmaceutical use. Such excipients can be solid, liquid, semisolid, or, in the case of an aerosol composition, gaseous.
“Pharmaceutically acceptable salts and esters” refers to salts and esters that are pharmaceutically acceptable and have the desired pharmacological properties. Such salts include, for example, salts that can be formed where acidic protons present in the compounds are capable of reacting with inorganic or organic bases. Suitable inorganic salts include, for example, those formed with the alkali metals or alkaline earth metals, e.g. sodium and potassium, magnesium, calcium, and aluminum. Suitable organic salts include, for example, those formed with organic bases such as the amine bases, e.g. ethanolamine, diethanolamine, triethanolamine, tromethamine, N methylglucamine, and the like. Pharmaceutically acceptable salts can also include acid addition salts formed from the reaction of basic moieties, such as amines, in the parent compound with inorganic acids (e.g. hydrochloric and hydrobromic acids) and organic acids (e.g. acetic acid, citric acid, maleic acid, and the alkane- and arene-sulfonic acids such as methanesulfonic acid and benzenesulfonic acid). Pharmaceutically acceptable esters include esters formed from carboxy, sulfonyloxy, and phosphonoxy groups present in the compounds, e.g. C1-6 alkyl esters. When there are two acidic groups present, a pharmaceutically acceptable salt or ester can be a mono-acid-mono-salt or ester or a di-salt or ester; and similarly where there are more than two acidic groups present, some or all of such groups can be salified or esterified. Compounds named in this invention can be present in unsalified or unesterified form, or in salified and/or esterified form, and the naming of such compounds is intended to include both the original (unsalified and unesterified) compound and its pharmaceutically acceptable salts and esters. Also, certain compounds named in this invention can be present in more than one stereoisomeric form, and the naming of such compounds is intended to include all single stereoisomers and all mixtures (whether racemic or otherwise) of such stereoisomers.
“Inhibitors,” “activators,” and “modulators” of expression or of activity are used to refer to inhibitory, activating, or modulating molecules, respectively, identified using in vitro and in vivo assays for expression or activity. Inhibitors of the present invention are compositions that, inhibit expression of PAI-1 or bind to, partially or totally block stimulation, decrease, prevent, delay activation, inactivate, desensitize, or down regulate the activity of PAI-1. Samples or assays comprising PAI-1 can be treated with a composition of the present invention and compared to control samples without a composition of the present invention. Control samples (untreated with compositions of the present invention) can be assigned a relative activity value of 100%. In certain embodiments, inhibition of PAI-1 is achieved when the activity value relative to the control is about 80% or less, optionally 50% or 25, 10%, 5% or 1%.
The terms “pharmaceutically acceptable”, “physiologically tolerable” and grammatical variations thereof, as they refer to compositions, carriers, diluents and reagents, are used interchangeably and represent that the materials are capable of administration to or upon a human without the production of undesirable physiological effects such as nausea, dizziness, gastric upset and the like which would be to a degree that would prohibit administration of the compound.
A “therapeutically effective amount” or “pharmaceutically effective amount” means the amount that, when administered to a subject, produces effects for which it is administered. For example, a “therapeutically effective amount,” when administered to a subject to inhibit PAI-1 activity, is sufficient to inhibit PAI-1 activity. A “therapeutically effective amount,” when administered to a subject for treating a disease, is sufficient to effect treatment for that disease.
Except when noted, the terms “subject” or “patient” are used interchangeably and refer to mammals such as human patients and non-human primates, as well as experimental animals such as rabbits, rats, and mice, and other animals. Accordingly, the term “subject” or “patient” as used herein means any mammalian patient or subject to which the compounds of the invention can be administered. In an exemplary embodiment of the present invention, to identify subject patients for treatment according to the methods of the invention, accepted screening methods are employed to determine risk factors associated with a targeted or suspected disease or condition or to determine the status of an existing disease or condition in a subject. These screening methods include, for example, conventional work-ups to determine risk factors that are associated with the targeted or suspected disease or condition. These and other routine methods allow the clinician to select patients in need of therapy using the methods and formulations of the present invention.
When any variable occurs more than one time in any constituent or in any formula, its definition in each occurrence is independent of its definition at every other occurrence. Combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.
B. Substituted Indoles
The present invention provides substituted indoles. Such compounds are preferably administered to inhibit PAI-1 expression or activity in a subject and, ultimately, to treat diseases or conditions associated with increased PAI-1 activity in a subject, e.g., a PAI-1 related disorder.
Substituted indoles include those of the following formula:
wherein:
X is a bond or C1-C6 alkylene
R1 and R2 are, independently, hydrogen, C1-C6 alkyl, C2-C7 alkenyl, C2-C7 alkynyl, C3-C8 cycloalkyl, C7-C11, bicycloalkyl, C6-C10 aryl, heterocyle, —C(═O)C1-C6 alkyl, —C(═O)aryl, —C(═O)heterocycle, —C(═O)N(R6)C1-C6 alkyl, —C(═O)N(R6)aryl, —C(═O)N(R6)heterocycle, —SO2—C1-C6 alkyl, —SO2-aryl, or —SO2-heterocycle;
or R1 and R2 together form a heterocycle;
R3 and R4 are independently, hydrogen, halogen, C1-C6 alkyl, C1-C3 perfluoroalkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, —C(═O)C1-C3 alkyl, —OH, —NH2, or —NO2;
R5 is hydrogen, C1-C6 alkyl, C2-C7 alkenyl, C2-C7 alkynyl, C3-C8 cycloalkyl, C7-C11 bicycloalkyl, C6-C10 aryl, or heterocycle;
A is hydrogen, C6-C10 aryl, or heterocycle; and
R6 is hydrogen, C1-C6 alkyl, halogen, C2-C7 alkenyl, C2-C7 alkynyl, C3-C8 cycloalkyl, aralkyl, C6-C10 aryl optionally substituted with R7, heterocycle optionally substituted with R7, hydroxy, C1-C6 alkoxy, aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO2H, —OCO2C1-C6 alkyl, —CO2C1-C6 alkyl, —CO2-aryl, —CO2(C1-C6 alkyl)aryl, —OCO2-aryl, —C(═O)NH2, —C(═O)NHOH, amino, C1-C6 alkylamino, dialkylamino of 1-6 carbons per alkyl moiety, —NHC(═O)NH—C1-C6 alkyl, —NHSO2—C1-C6 alkyl, —NHSO2-aryl, or —NHSO2-heterocycle.
Compounds of the present invention also include prodrugs, stereoisomers, or pharmaceutically acceptable salts or ester forms of Formula 1.
For use in the present invention, R1 and R2 can be hydrogen, C1-C6 alkyl, C2-C7 alkenyl, C2-C7 alkynyl, C3-C8 cycloalkyl, C7-C11 bicycloalkyl, C6-C10 aryl, heterocycle, —C(═O)C1-C6 alkyl, —C(═O)aryl, —C(═O)heterocycle, —C(═O)N(R6)C1-C6 alkyl, —C(═O)N(R6)aryl, —C(═O)N(R6)heterocycle, —SO2—C1-C6 alkyl, —SO2-aryl, or —SO2-heterocycle. In certain exemplary embodiments of the present invention, R1 or R2 are, independently, hydrogen, SO2-alkyl or SO2-aryl optionally substituted with —OCF3, aryl, or alkyl. In certain preferred embodiments, R1 is hydrogen and R2 is SO2-methyl or SO2-phenyl optionally substituted with —OCF3, phenyl, or tert-butyl. In such embodiments, R3, R4, R5, R6, X, and A are as defined herein.
In some exemplary embodiments of the present invention, R1 and R2 together form a heterocycle. For example, in certain embodiments, R1 and R2 together form pyrrole. In such embodiments, R3, R4, R5, R6, X, and A are as defined herein.
R3 and R4 can be hydrogen, halogen, C1-C6 alkyl, C1-C3 perfluoroalkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, —C(═O)C1-C3 alkyl, —OH, —NH2, or —NO2. In some embodiments of the present invention, R3 and R4 are, independently, hydrogen, or alkyl optionally substituted with halogen, —CN, or alkoxy. In certain preferred embodiments, R3 and R4 are hydrogen. In such embodiments, R1, R2, R5, R6, X, and A are as defined herein.
R5 can be hydrogen, C1-C6 alkyl, C2-C7 alkenyl, C2-C7 alkynyl, C3-C8 cycloalkyl, C7-C11 bicycloalkyl, C6-C10 aryl, or heterocycle. In certain exemplary embodiments of the present invention, R5 is hydrogen. In such embodiments, R1, R2, R3, R4, R6, X, and A are as defined herein.
R6 can be C1-C6 alkyl, halogen, C2-C7 alkenyl, C2-C7 alkynyl, C3-C8 cycloalkyl, aralkyl, C6-C10 aryl optionally substituted with R7, heterocycle optionally substituted with R7, hydroxy, C1-C6 alkoxy, aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO2H, —OCO2C1-C6 alkyl, —CO2C1-C6 alkyl, —CO2-aryl, —CO2(C1-C6 alkyl)aryl, —OCO2-aryl, —C(═O)NH2, —C(═O)NHOH, amino, C1-C6 alkylamino, dialkylamino of 1-6 carbons per alkyl moiety, —NHC(═O)NH—C1-C6 alkyl, —NHSO2—C1-C6 alkyl, —NHSO2-aryl, or —NHSO2-heterocycle.
A can be hydrogen, C6-C10 aryl, or heterocycle. In certain exemplary embodiments of the present invention, A is hydrogen, phenyl, or thiophene. In such embodiments, R1, R2, R3, R4, R5, R6, and X are as defined herein.
Preferred compounds of the present invention include those wherein
X is a bond or unsubstituted C1-C6 alkylene;
R1 and R2 are, independently, hydrogen, C1-C6 alkyl, C2-C7 alkenyl, C2-C7 alkynyl, C3-C8 cycloalkyl, C7-C11 bicycloalkyl, C6-C10 aryl, heterocycle, unsubstituted —C(═O)C1-C6 alkyl, unsubstituted —C(═O)aryl, unsubstituted —C(═O)heterocycle, unsubstituted —C(═O)N(R6)C1-C6 alkyl, unsubstituted —C(═O)N(R6)aryl, unsubstituted —C(═O)N(R6)heterocycle, unsubstituted —SO2—C1-C6 alkyl, unsubstituted —SO2-aryl, or unsubstituted —SO2-heterocycle;
wherein said C1-C6 alkyl, C2-C7 alkenyl, C2-C7 alkynl, C3-C8 cycloalkyl, C7-C11 bicycloalkyl, C6-C10 aryl, heterocycle groups are optionally substituted with unsubstituted C1-C6 alkyl, halogen, unsubstituted C2-C7 alkenyl, unsubstituted C2-C7 alkynyl, unsubstituted C3-C8 cycloalkyl, unsubstituted aralkyl, hydroxy, unsubstituted C1-C6 alkoxy, unsubstituted aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO2H, unsubstituted —OCO2C1-C6 alkyl, unsubstituted CO2C1-C6 alkyl, unsubstituted —CO2-aryl, unsubstituted —CO2(C1-C6 alkyl)aryl, unsubstituted —OCO2-aryl, —C(═O)NH2, —C(═O)NHOH, unsubstituted amino, unsubstituted alkylamino, unsubstituted dialkylamino, unsubstituted —NHC(═O)NH—C1-C6 alkyl, unsubstituted —NHSO2—C1-C6 alkyl, unsubstituted —NHSO2-aryl, unsubstituted —NHSO2-heterocycle, aryl (optionally substituted with halogen, unsubstituted C1-C6 alkoxy, unsubstituted C1-C6 alkyl, unsubstituted C2-C7 alkenyl, unsubstituted C2-C7 alkynyl, hydroxy, unsubstituted —C(═O)C1-C7 alkyl, unsubstituted —SO2—C1-C6 alkyl, unsubstituted —CO2—C1-C6 alkyl, or unsubstituted alkoxycarbonylalkyl) or heterocycle (optionally substituted with halogen, unsubstituted C1-C6 alkoxy, unsubstituted C1-C6 alkyl, unsubstituted C2-C7 alkenyl, unsubstituted C2-C7 alkynyl, hydroxy, unsubstituted —C(═O)C1-C7 alkyl, unsubstituted —SO2—C1-C6 alkyl, unsubstituted —CO2—C1-C6 alkyl, or unsubstituted alkoxycarbonylalkyl);
or R1 and R2 together form a heterocycle optionally substituted with unsubstituted C1-C6 alkyl, halogen, unsubstituted C2-C7 alkenyl, unsubstituted C2-C7 alkynyl, unsubstituted C3-C8 cycloalkyl, unsubstituted aralkyl, hydroxy, unsubstituted C1-C6 alkoxy, unsubstituted aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO2H, unsubstituted —OCO2C1-C6 alkyl, unsubstituted CO2C1-C6 alkyl, unsubstituted —CO2-aryl, unsubstituted —CO2(C1-C6 alkyl)aryl, unsubstituted —OCO2-aryl, —C(═O)NH2, —C(═O)NHOH, unsubstituted amino, unsubstituted alkylamino, unsubstituted dialkylamino, unsubstituted —NHC(═O)NH—C1-C6 alkyl, unsubstituted —NHSO2—C1-C6 alkyl, unsubstituted —NHSO2-aryl, unsubstituted —NHSO2-heterocycle, aryl (optionally substituted with halogen, unsubstituted C1-C6 alkoxy, unsubstituted C1-C6 alkyl, unsubstituted C2-C7 alkenyl, unsubstituted C2-C7 alkynyl, hydroxy, unsubstituted —C(═O)C1-C7 alkyl, unsubstituted —SO2—C1-C6 alkyl, unsubstituted —CO2—C1-C6 alkyl, or unsubstituted alkoxycarbonylalkyl) or heterocycle (optionally substituted with halogen, unsubstituted C1-C6 alkoxy, unsubstituted C1-C6 alkyl, unsubstituted C2-C7 alkenyl, unsubstituted C2-C7 alkynyl, hydroxy, unsubstituted —C(═O)C1-C7 alkyl, unsubstituted —SO2—C1-C6 alkyl, unsubstituted —CO2—C1-C6 alkyl, or unsubstituted alkoxycarbonylalkyl);
R3 and R4 are independently, hydrogen, halogen, unsubstituted C1-C3 perfluoroalkyl, unsubstituted C1-C6 alkoxy, unsubstituted C3-C8 cycloalkyl, unsubstituted —C(═O)C1-C3 alkyl, —OH, —NH2, —NO2 or C1-C6 alkyl optionally substituted with halogen, —CN, or C1-C3 alkoxy;
R5 is hydrogen, C1-C6 alkyl, C2-C7 alkenyl, C2-C7 alkynyl, C3-C8 cycloalkyl, C7-C11 bicycloalkyl, C6-C10 aryl, or heterocycle
wherein said wherein said C1-C6 alkyl, C2-C7 alkenyl, C2-C7 alkynyl, C3-C8 cycloalkyl, C7-C11 bicycloalkyl, C6-C10 aryl, or heterocycle groups are optionally substituted with unsubstituted C1-C6 alkyl, halogen, unsubstituted C2-C7 alkenyl, unsubstituted C2-C7 alkynyl, unsubstituted C3-C8 cycloalkyl, unsubstituted aralkyl, hydroxy, unsubstituted C1-C6 alkoxy, unsubstituted aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO2H, unsubstituted —OCO2C1-C6 alkyl, unsubstituted CO2C1-C6 alkyl, unsubstituted —CO2-aryl, unsubstituted —CO2(C1-C6 alkyl)aryl, unsubstituted —OCO2-aryl, —C(═O)NH2, —C(═O)NHOH, unsubstituted amino, unsubstituted alkylamino, unsubstituted dialkylamino, unsubstituted —NHC(═O)NH—C1-C6 alkyl, unsubstituted —NHSO2—C1-C6 alkyl, unsubstituted —NHSO2-aryl, unsubstituted —NHSO2-heterocycle, aryl (optionally substituted with halogen, unsubstituted C1-C6 alkoxy, unsubstituted C1-C6 alkyl, unsubstituted C2-C7 alkenyl, unsubstituted C2-C7 alkynyl, hydroxy, unsubstituted —C(═O)C1-C7 alkyl, unsubstituted —SO2—C1-C6 alkyl, unsubstituted —CO2—C1-C6 alkyl, or unsubstituted alkoxycarbonylalkyl) or heterocycle (optionally substituted with halogen, unsubstituted C1-C6 alkoxy, unsubstituted C1-C6 alkyl, unsubstituted C2-C7 alkenyl, unsubstituted C2-C7 alkynyl, hydroxy, unsubstituted —C(═O)C1-C7 alkyl, unsubstituted —SO2—C1-C6 alkyl, unsubstituted —CO2—C1-C6 alkyl, or unsubstituted alkoxycarbonylalkyl);
A is hydrogen, C6-C10 aryl, or heterocycle
wherein said aryl or heterocycle groups are optionally substituted with unsubstituted C1-C6 alkyl, halogen, unsubstituted C2-C7 alkenyl, unsubstituted C2-C7 alkynyl, unsubstituted C3-C8 cycloalkyl, unsubstituted aralkyl, hydroxy, unsubstituted C1-C6 alkoxy, unsubstituted aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO2H, unsubstituted —OCO2C1-C6 alkyl, unsubstituted CO2C1-C6 alkyl, unsubstituted —CO2-aryl, unsubstituted —CO2(C1-C6 alkyl)aryl, unsubstituted —OCO2-aryl, —C(═O)NH2, —C(═O)NHOH, unsubstituted amino, unsubstituted alkylamino, unsubstituted dialkylamino, unsubstituted —NHC(═O)NH—C1-C6 alkyl, unsubstituted —NHSO2—C1-C6 alkyl, unsubstituted —NHSO2-aryl, unsubstituted —NHSO2-heterocycle, aryl (optionally substituted with halogen, unsubstituted C1-C6 alkoxy, unsubstituted C1-C6 alkyl, unsubstituted C2-C7 alkenyl, unsubstituted C2-C7 alkynyl, hydroxy, unsubstituted —C(═O)C1-C7 alkyl, unsubstituted —SO2—C1-C6 alkyl, unsubstituted —CO2—C1-C6 alkyl, or unsubstituted alkoxycarbonylalkyl) or heterocycle (optionally substituted with halogen, unsubstituted C1-C6 alkoxy, unsubstituted C1-C6 alkyl, unsubstituted C2-C7 alkenyl, unsubstituted C2-C7 alkynyl, hydroxy, unsubstituted —C(═O)C1-C7 alkyl, unsubstituted —SO2—C1-C6 alkyl, unsubstituted —CO2—C1-C6 alkyl, or unsubstituted alkoxycarbonylalkyl);
and R6 is hydrogen, unsubstituted C1-C6 alkyl, halogen, unsubstituted C2-C7 alkenyl, unsubstituted C2-C7 alkynyl, unsubstituted C3-C8 cycloalkyl, unsubstituted aralkyl, hydroxy, unsubstituted C1-C6 alkoxy, unsubstituted aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO2H, unsubstituted —OCO2C1-C6 alkyl, unsubstituted CO2C1-C6 alkyl, unsubstituted —CO2-aryl, unsubstituted —CO2(C1-C6 alkyl)aryl, unsubstituted —OCO2-aryl, —C(═O)NH2, —C(═O)NHOH, unsubstituted amino, unsubstituted alkylamino, unsubstituted dialkylamino, unsubstituted —NHC(═O)NH—C1-C6 alkyl, unsubstituted —NHSO2—C1-C6 alkyl, unsubstituted —NHSO2-aryl, unsubstituted —NHSO2-heterocycle, aryl (optionally substituted with halogen, unsubstituted C1-C6 alkoxy, unsubstituted C1-C6 alkyl, unsubstituted C2-C7 alkenyl, unsubstituted C2-C7 alkynyl, hydroxy, unsubstituted —C(═O)C1-C7 alkyl, unsubstituted —SO2—C1-C6 alkyl, unsubstituted —CO2—C1-C6 alkyl, or unsubstituted alkoxycarbonylalkyl) or heterocycle (optionally substituted with halogen, unsubstituted C1-C6 alkoxy, unsubstituted C1-C6 alkyl, unsubstituted C2-C7 alkenyl, unsubstituted C2-C7 alkynyl, hydroxy, unsubstituted —C(═O)C1-C7 alkyl, unsubstituted —SO2—C1-C6 alkyl, unsubstituted —CO2—C1-C6 alkyl, or unsubstituted alkoxycarbonylalkyl).
In certain embodiments of the present invention, substituted indoles include the following compounds:
wherein R3, R4, X, A, and n are as defined above, and
Y is —NHSO2—C1-C6 alkyl, —NHSO2-aryl, —NHSO2-heterocycle, pyrrole, —NH-aryl, or —NH-heterocycle.
In representative embodiments of the present invention, the pyrrole group is substituted by C1-C6 alkyl, halogen, C2-C7 alkenyl, C2-C7 alkynyl, C3-C8 cycloalkyl, aralkyl wherein n is from 1 to 6, aryl, heterocycle, hydroxy, C1-C6 alkoxy, aryl-oxy, oxo (═O), —CN, —C(═O)H, —CO2H, —OCO2C1-C6 alkyl, —CO2C1-C6 alkyl, —CO2-aryl, —CO2(C1-C6 alkyl)aryl, —OCO2-aryl, —C(═O)NH2, —C(═O)NHOH, amino, alkylamino, dialkylamino, —NHC(═O)NH—C1-C6 alkyl, —NHSO2—C1-C6 alkyl, —NHSO2-aryl, or —NHSO2-heterocycle.
Exemplary substituted 1-aralkyl or 1-aryl-1H-indoles of the present invention include, but are not limited to, 1-[4-({[4-(trifluoromethoxy)phenyl]sulfonyl}amino)phenyl]-1H-indole-2-carboxylic acid or a pharmaceutically acceptable salt or ester form thereof; 1-[4-(1H-pyrrol-1-yl)phenyl]-1H-indole-2-carboxylic acid or a pharmaceutically acceptable salt or ester form thereof; 1-{4-[(1,1′-biphenyl-4-ylsulfonyl)amino]benzyl}-1H-indole-2-carboxylic acid or a pharmaceutically acceptable salt or ester form thereof; 1-(4-{[(4-tert-butylphenyl)sulfonyl]amino}benzyl)-1H-indole-5-carboxylic acid or a pharmaceutically acceptable salt or ester form thereof; 1-{4-[(1,1′-biphenyl-4-ylsulfonyl)amino]benzyl}-1H-indole-5-carboxylic acid or a pharmaceutically acceptable salt or ester form thereof; 1-(4-{[(4-tert-butylphenyl)sulfonyl]amino}benzyl)-1H-indole-3-carboxylic acid or a pharmaceutically acceptable salt or ester form thereof; 1-{4-[(1,1′-biphenyl-4-ylsulfonyl)amino]benzyl}-1H-indole-3-carboxylic acid or a pharmaceutically acceptable salt or ester form thereof; 1-(4-{[(4-tert-butylphenyl)sulfonyl]amino}benzyl)-1H-indole-4-carboxylic acid or a pharmaceutically acceptable salt or ester form thereof; 1-{4-[(1,1′-biphenyl-4-ylsulfonyl)amino]benzyl}-1H-indole-4-carboxylic acid or a pharmaceutically acceptable salt or ester form thereof; 1-{4-[(1,1′-biphenyl-4-ylsulfonyl)amino]benzyl}-1H-indole-6-carboxylic acid or a pharmaceutically acceptable salt or ester form thereof; 3-phenyl-1-{4-[(phenylsulfonyl)amino]benzyl}-1H-indole-2-carboxylic acid or a pharmaceutically acceptable salt or ester form thereof; 1-{4-[(methylsulfonyl)amino]benzyl}-3-phenyl-1H-indole-2-carboxylic acid or a pharmaceutically acceptable salt or ester form thereof; 1-{4-[(phenylsulfonyl)amino]benzyl}-3-thien-2-yl-1H-indole-2-carboxylic acid or a pharmaceutically acceptable salt or ester form thereof; 1-{4-[(methylsulfonyl)amino]benzyl}-3-thien-2-yl-1H-indole-2-carboxylic acid or a pharmaceutically acceptable salt or ester form thereof; 1-[4-(2,5-dimethyl-1H-pyrrol-1-yl)benzyl]-3-phenyl-1H-indole-2-carboxylic acid or a pharmaceutically acceptable salt or ester form thereof; 1-(4-anilinobenzyl)-3-phenyl-1H-indole-2-carboxylic acid or a pharmaceutically acceptable salt or ester form thereof; 1-(4-anilinobenzyl)-3-thien-2-yl-1H-indole-2-carboxylic acid or a pharmaceutically acceptable salt or ester form thereof.
The present invention also provides compositions comprising substituted indoles, including those compounds of formulas 1 and 2 or a stereoisomer or pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, excipients, or diluents. Such compositions include pharmaceutical compositions for treating or controlling disease states or conditions associated with increased PAI-1 activity. In certain embodiments, the compositions comprise mixtures of one or more substituted indoles.
Certain of the compounds of formulas 1 and 2 contain stereogenic carbon atoms or other chiral elements and thus give rise to stereoisomers, including enantiomers and diastereomers. The present invention includes all of the stereoisomers of formulas 1 and 2, as well as mixtures of the stereoisomers. Throughout this application, the name of the product, where the absolute configuration of an asymmetric center is not indicated, is intended to embrace the individual stereoisomers as well as mixtures of stereoisomers.
Where an enantiomer is preferred, it can, in some embodiments, be provided substantially free of the corresponding enantiomer. Thus, an enantiomer substantially free of the corresponding enantiomer refers to a compound that is isolated or separated via separation techniques or prepared free of the corresponding enantiomer. “Substantially free,” as used herein, means that the compound is made up of a significantly greater proportion of one enantiomer. In preferred embodiments, the compound is made up of at least about 90% by weight of a preferred enantiomer. In other embodiments of the invention, the compound is made up of at least about 99% by weight of a preferred enantiomer. Preferred enantiomers can be isolated from racemic mixtures by any method known to those skilled in the art, including high performance liquid chromatography (HPLC) and the formation and crystallization of chiral salts, or preferred enantiomers can be prepared by methods described herein. Methods for the preparation of preferred enantiomers are described, for example, in Jacques, et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen, S. H., et al., Tetrahedron 33:2725 (1977); Eliel, E. L. Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, S. H. Tables of Resolving Agents and Optical Resolutions p. 268 (E. L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, Ind. 1972).
Exemplary salt forms of the compounds herein include, but are not limited to, sodium salts and potassium salts. Other exemplary salt forms of these compounds include, but are not limited to, those formed with pharmaceutically acceptable inorganic and organic bases or acids known in the art. The acids include, for example, acetic, propionic, lactic, citric, tartaric, succinic, fumaric, maleic, malonic, mandelic, malic, phthalic, hydrochloric, hydrobromic, phosphoric, nitric, sulfuric, methanesulfonic, napthalenesulfonic, benzenesulfonic, toluenesulfonic, camphorsulfonic, and similarly known acceptable aids when a compound of this invention contains a basic moiety. Salt forms prepared using inorganic bases include hydroxides, carbonates or bicarbonates of the therapeutically acceptable alkali metals or alkaline earth metals, such as sodium potassium, magnesium, calcium and 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 triethanolamine. Exemplary salts also include 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 can also be formed, such as tetralkyl forms, such as tetramethyl forms, alkyl-alkanol forms, such as methyl-triethanol or trimethyl-monoethanol forms, and cyclic ammonium salt forms, such as N-methylpyridinium, N-methyl-N-(2-hydroxyethyl)-morpholinium, N,N-di-methylmorpholinium, N-methyl-N-(2-hydroxyethyl)-morpholinium, or N,N-dimethyl-piperidinium salt forms. These salt forms can be prepared using the acidic compound(s) of formulas 1 or 2 and procedures known in the art.
Exemplary ester forms of the compounds of this invention include, but are not limited to, straight chain alkyl esters having from 1 to 6 carbon atoms or branched chain alkyl groups containing 1 to 6 carbon atoms, including methyl, ethyl, propyl, butyl, 2-methylpropyl and 1,1-dimethylethyl esters, cycloalkyl esters, alkylaryl esters, benzyl esters, and the like. Other exemplary esters include, but are not limited to, those of the formula —COOR12 wherein R12 is selected from the formula:
wherein R8, R9, R10, and R11 are independently selected from hydrogen, alkyl of from 1 to 10 carbon atoms, aryl of 6 to 12 carbon atoms, arylalkyl of from 6 to 12 carbon atoms; heteroaryl or alkylheteroaryl wherein the heteroaryl ring is bound by an alkyl chain of from 1 to 6 carbon atoms.
Preferred compounds of the present invention inhibit PAI-1 activity. Accordingly, the compounds can be used for the treatment, including prevention, inhibition, and/or amelioration of PAI-1 related disorders in a subject, including, for example, in the treatment of noninsulin dependent diabetes mellitus, in the treatment of cardiovascular disease, and in the treatment of thrombotic events associated with coronary artery and cerebrovascular disease. Using the methods of the present invention, a skilled medical practitioner will know how to administer substituted indoles, including those represented by formulas 1 and 2, to a subject suffering from any of the diseases associated with increased PAI-1 activity or expression, e.g., diabetes or cardiovascular disease, in order to effect treatment for that disease.
In one exemplary embodiment, substituted indoles are administered to a subject in order to treat disease processes involving 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 thrombosis, cerebral thrombosis, thromboembolic complications of surgery (such as joint or hip replacement), and peripheral arterial occlusion.
Any disease or condition that is associated with increased PAI-1 activity or expression in a subject can be treated using substituted indoles of the present invention. Exemplary diseases and conditions include stroke, e.g., stroke associated with or resulting from atrial fibrillation; diseases associated with extracellular matrix accumulation including, but not limited to, renal fibrosis, chronic obstructive pulmonary disease, polycystic ovary syndrome, restenosis, renovascular disease, and organ transplant rejection; diseases associated with neoangiogenesis, including, but not limited to, diabetic retinopathy; Alzheimer's disease, e.g., by increasing or normalizing levels of plasmin concentration in a subject; myelofibrosis with myeloid metaplasia, e.g., by regulating stromal cell hyperplasia and increases in extracellular matrix proteins; diabetic nephropathy and renal dialysis associated with nephropathy; malignancies or cancers, including, but not limited to, leukemia, breast cancer and ovarian cancer; tumors, including, but not limited to, liposarcomas and epithelial tumors; septicemia; obesity; insulin resistance; proliferative diseases, including, but not limited to, psoriasis; conditions associated with abnormal coagulation homeostasis; low grade vascular inflammation; cerebrovascular diseases; hypertension; dementia; osteoporosis; arthritis; respiratory diseases, such as asthma; heart failure; arrhythmia; angina, including, but not limited to, angina pectoris; atherosclerosis and sequelae; kidney failure; multiple sclerosis; osteoporosis; osteopenia; dementia; peripheral vascular disease; peripheral arterial disease; acute vascular syndromes; microvascular diseases including, but not limited to, nephropathy, neuropathy, retinopathy and nephrotic syndrome; hypertension; Type I and II diabetes and related diseases; hyperglycemia; hyperinsulinemia; malignant lesions; premalignant lesions; gastrointestinal malignancies; coronary heart disease, including, but not limited to, primary and secondary prevention of myocardial infarction, stable and unstable angina, primary prevention of coronary events, and secondary prevention of cardiovascular events; and inflammatory diseases, including, but not limited to, septic shock and the vascular damage associated with infections.
The compounds of the present invention can also be administered to a subject in combination with a second therapeutic agent, including, but not limited to, prothrombolytic, fibrinolytic, and anticoagulant agents, or in conjunction with other therapies, for example, protease inhibitor-containing highly active antiretroviral therapy (HAART) for the treatment of diseases which originate from fibrinolytic impairment and hyper-coagulability of HIV-1 infected patients. In certain embodiments, the compounds of the present invention can be administered in conjunction with and/or following processes or procedures involving maintaining blood vessel patency, including, but not limited to, vascular surgery, vascular graft and stent patency, organ, tissue and cell implantation and transplantation. The compounds of the present invention can also be used for the treatment of blood and blood products used in dialysis, blood storage in the fluid phase, especially ex vivo platelet aggregation. The compounds of the present invention can also be administered to a subject as a hormone replacement agent or to reduce inflammatory markers or C-reactive protein. The compounds can be administered to improve coagulation homeostasis, to improve endothelial function, or as a topical application for wound healing, e.g., the prevention of scarring. The compounds of the present invention can be administered to a subject in order to reduce the risk of undergoing a myocardial revascularization procedure. The present compounds can also be added to human plasma during the analysis of blood chemistry in hospital settings to determine the fibrinolytic capacity thereof. In certain embodiments, the compounds of the present invention can be used as imaging agents for the identification of metastatic cancers.
C. Synthesis of Substituted Indoles
Compounds of the present invention can be prepared by those skilled in the art of organic synthesis employing conventional methods that utilize readily available reagents and starting materials. Representative compounds of the present invention can be prepared using the following synthetic schemes. The skilled practitioner will know how to make use of variants of these process steps, which in themselves are well known in the art. In the following reaction schemes, R1, R2, R3, R4, R5, R6, and Ar (aryl) are as defined above for Formula 1.
The 1H-indole 1 can be alkylated with substituted nitrobenzyl halides in the presence of a base, such as K2CO3, Cs2CO3, KOH or NaH, in an inert solvent, such as THF, dioxane, pyridine, DMF, NMP, or DMSO, at −40 to 100° C. The resulting nitro intermediate 2 can be reduced to the aniline 3 upon treatment with Raney® nickel in a mixture of hydrazine and ethanol at temperature of 0 to 40° C. Compound 3 is then converted to sulfonamide 4, wherein R13 is alkyl, aryl, or Het, upon treatment with a sulfonyl halide, preferably alkyl, aryl, or heterocycle sulfonyl chloride, and a base such as N,N-diisopropylethylamine in an anhydrous solvent such as dichloromethane for 0.5 to 24 hours at temperature of 0 to 40° C. Condensation of aniline 3 with an excess of 1,4-dicarbonyl compounds, such as acetonylacetone, utilizing a Dean-Stark trap for azeotropic water removal leads to pyrrolylindole 6 (Paal-Knorr synthesis), wherein R14 and R15 are independently hydrogen, alkyl, aryl, or heterocycle. A variant of the Paal-Knorr synthesis which may be applicable to the synthesis of 1-substituted pyrroles is the reaction of aniline 3 with an excess of the cyclic acetal 2,5-dimethoxytetrahydrofuran. The aniline 3 can also be converted to diaryl amine 8 with boronic acid in the presence of a base, preferably 2,6-lutidine, an additive such as myristic acid, and Cu(OAc)2 in an inert solvent such as toluene at room temperature. The final acids 5, 7 or 9 are obtained via a basic hydrolysis of the corresponding ester 4, 6 or 8. In this reaction lithium hydroxide, sodium hydroxide, potassium hydroxide and the like can be used as a base, and water or a mixture of water with methanol, ethanol, dioxane and the like can be used as a solvent. The final products can be purified by recrystallization, trituration, preparative thin layer chromatography, flash column chromatography on silica gel, or high performance liquid chromatography. Purification of intermediates can be achieved in the same manner. A salt is optionally produced by the addition of an acid or base, such as hydrogen chloride gas or hydrochloric acid.
Aryl indole derivatives 12 are also readily prepared by methods described in the literature or known to those skilled in the art. For example, as shown in Scheme II, the 1H-indole 1 can be arylated with substituted 4-fluoronitrobenzene in the presence of a base, such Cs2CO3, in an inert solvent, such as DMF, at 0 to 40° C. The resulting nitro intermediate 10 can be reduced to the desired aniline 11 upon treatment with Raney® nickel in a mixture of hydrazine and ethanol at temperature of 0 to 40° C. The aniline 11 can be converted to N-aryl indole derivatives 12, such as sulfonamides, pyrroles, and diaryl amines as described in Scheme 1.
The C3 aryl or heterocycle substituted indole derivative 16, wherein R16 is aryl or heterocycle, can be readily prepared as shown in Scheme III. Substituted indole 1 is first halogenated on the indole C3 position with known halogenation agents such as chlorine, bromine, N-chlorosuccinimide, N-bromosuccinimide and the like in a solvent such as carbon tetrachloride, chloroform, dimethylformamide (DMF), or N-methylpyrrolidinone (NMP) at temperatures of 0 to 30° C. Halo in compound 13 is chlorine, bromine or iodine. The C3—R16 group can be introduced by a Suzuki coupling reaction with boronic acid in the presence of a base, preferably Na2CO3, and Pd(PPh3)4 in an inert solvent such as toluene at 80 to 100° C. The 1H-indole intermediate 14 can then be alkylated or arylated to afford the desired intermediate 15 which cab to converted to the final product 16 as described in Scheme 1 and Scheme 2.
D. Substituted Indoles as Pharmaceutical Compositions
The present invention provides substituted indoles as pharmaceuticals. In a preferred embodiment, substituted indoles are formulated as pharmaceuticals to treat diseases associated with increased PAI-1 activity, e.g., by inhibiting PAI-1 activity in a subject.
In general, substituted indoles can be administered as pharmaceutical compositions by any method known in the art for administering therapeutic drugs including oral, buccal, topical, systemic (e.g., transdermal, intranasal, or by suppository), or parenteral (e.g., intramuscular, subcutaneous, or intravenous injection). Compositions can take the form of tablets, pills, capsules, semisolids, powders, sustained release formulations, solutions, suspensions, emulsions, syrups, elixirs, aerosols, or any other appropriate compositions; and comprise at least one compound of this invention in combination with at least one pharmaceutically acceptable excipient. Suitable excipients are well known to persons of ordinary skill in the art, and they, and the methods of formulating the compositions, can be found in such standard references as Alfonso A R: Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton Pa., 1985. Suitable liquid carriers, especially for injectable solutions, include water, aqueous saline solution, aqueous dextrose solution, and glycols. In some embodiments of the present invention, substituted indoles suitable for use in the practice of this invention will be administered either singly or in combination with at least one other compound of this invention. Substituted indoles suitable for use in the practice of the present invention can also be administered with at least one other conventional therapeutic agent for the disease being treated.
Aqueous suspensions of the invention can contain a substituted indole in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients can include a suspending agent, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia, and dispersing or wetting agents such as a naturally occurring phosphatide (e.g., lecithin), a condensation product of an alkylene oxide with a fatty acid (e.g., polyoxyethylene stearate), a condensation product of ethylene oxide with a long chain aliphatic alcohol (e.g., heptadecaethylene oxycetanol), a condensation product of ethylene oxide with a partial ester derived from a fatty acid and a hexitol (e.g., polyoxyethylene sorbitol mono-oleate), or a condensation product of ethylene oxide with a partial ester derived from fatty acid and a hexitol anhydride (e.g., polyoxyethylene sorbitan mono-oleate). The aqueous suspension can also contain one or more preservatives such as ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose, aspartame or saccharin. Formulations can be adjusted for osmolarity.
Oil suspensions can be formulated by suspending a substituted indole in a vegetable oil, such as arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin; or a mixture of these. The oil suspensions can contain a thickening agent, such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents can be added to provide a palatable oral preparation, such as glycerol, sorbitol or sucrose. These formulations can be preserved by the addition of an antioxidant such as ascorbic acid. As an example of an injectable oil vehicle, see Minto, J. Pharmacol. Exp. Ther. 281:93-102, 1997. The pharmaceutical formulations of the invention can also be in the form of oil-in-water emulsions. The oily phase can be a vegetable oil or a mineral oil, described above, or a mixture of these. Suitable emulsifying agents include naturally-occurring gums, such as gum acacia and gum tragacanth, naturally occurring phosphatides, such as soybean lecithin, esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan mono-oleate, and condensation products of these partial esters with ethylene oxide, such as polyoxyethylene sorbitan mono-oleate. The emulsion can also contain sweetening agents and flavoring agents, as in the formulation of syrups and elixirs. Such formulations can also contain a demulcent, a preservative, or a coloring agent.
The compound of choice, alone or in combination with other suitable components, can be made into aerosol formulations (i.e., they can be “nebulized”) to be administered via inhalation. Aerosol formulations can be placed into pressurized acceptable propellants, such as dichlorodifluoromethane, propane, nitrogen, and the like.
Formulations suitable for parenteral administration, such as, for example, by intraarticular (in the joints), intravenous, intramuscular, intradermal, intraperitoneal, and, subcutaneous routes, include aqueous and non-aqueous, isotonic sterile injection solutions, which can contain antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions that can include suspending agents, solubilizers, thickening agents, stabilizers, and preservatives. Among the acceptable vehicles and solvents that can be employed are water and Ringer's solution, an isotonic sodium chloride. In addition, sterile fixed oils can conventionally be employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid can likewise be used in the preparation of injectables. These solutions are sterile and generally free of undesirable matter. 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. Dispersions of the finely divided compounds can be made-up in aqueous starch or sodium carboxymethyl cellulose solution, or in suitable oil, such as arachis oil. These formulations can be sterilized by conventional, well known sterilization techniques. The formulations can contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions such as pH adjusting and buffering agents, toxicity adjusting agents, e.g., sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate and the like. The concentration of substituted indoles in these formulations can vary widely, and will be selected primarily based on fluid volumes, viscosities, body weight, and the like, in accordance with the particular mode of administration selected and the patient's needs. For IV administration, the formulation can be a sterile injectable preparation, such as a sterile injectable aqueous or oleaginous suspension. This suspension can be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation can also be a sterile injectable solution or suspension in a nontoxic parenterally-acceptable diluent or solvent, such as a solution of 1,3-butanediol. The formulations of commends can be presented in unit-dose or multi-dose sealed containers, such as ampules and vials.
Injection solutions and suspensions can be prepared from sterile powders, granules, and tablets of the kind previously described.
Substituted indoles suitable for use in the practice of this invention can be administered orally. The amount of a compound of the present invention in the composition can vary widely depending on the type of composition, size of a unit dosage, kind of excipients, and other factors well known to those of ordinary skill in the art. In general, the final composition can comprise from, for example, 0.000001 percent by weight (% w) to 10% w of the substituted substituted indole, preferably 0.00001% w to 1% w, with the remainder being the excipient or excipients.
Pharmaceutical formulations for oral administration can be formulated using pharmaceutically acceptable carriers well known in the art in dosages suitable for oral administration. Such carriers enable the pharmaceutical formulations to be formulated in unit dosage forms as tablets, pills, powder, dragees, capsules, liquids, lozenges, gels, syrups, slurries, suspensions, etc. suitable for ingestion by the patient. Formulations suitable for oral administration can consist of (a) liquid solutions, such as an effective amount of the packaged nucleic acid suspended in diluents, such as water, saline or PEG 400; (b) capsules, sachets or tablets, each containing a predetermined amount of the active ingredient, as liquids, solids, granules or gelatin; (c) suspensions in an appropriate liquid; and (d) suitable emulsions.
Pharmaceutical preparations for oral use can be obtained through combination of the compounds of the present invention with a solid excipient, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable additional compounds, if desired, to obtain tablets or dragee cores. Suitable solid excipients are carbohydrate or protein fillers and include, but are not limited to sugars, including lactose, sucrose, mannitol, or sorbitol; starch from corn, wheat, rice, potato, or other plants; cellulose such as methyl cellulose, hydroxymethyl cellulose, hydroxypropylmethyl-cellulose or sodium carboxymethylcellulose; and gums including arabic and tragacanth; as well as proteins such as gelatin and collagen. If desired, disintegrating or solubilizing agents can be added, such as the cross-linked polyvinyl pyrrolidone, agar, alginic acid, or a salt thereof, such as sodium alginate. Tablet forms can include one or more of lactose, sucrose, mannitol, sorbitol, calcium phosphates, corn starch, potato starch, microcrystalline cellulose, gelatin, colloidal silicon dioxide, talc, magnesium stearate, stearic acid, and other excipients, colorants, fillers, binders, diluents, buffering agents, moistening agents, preservatives, flavoring agents, dyes, disintegrating agents, and pharmaceutically compatible carriers. Lozenge forms can comprise the active ingredient in a flavor, e.g., sucrose, as well as pastilles comprising the active ingredient in an inert base, such as gelatin and glycerin or sucrose and acacia emulsions, gels, and the like containing, in addition to the active ingredient, carriers known in the art.
The substituted indoles of the present invention can also be administered in the form of suppositories for rectal administration of the drug. These formulations can be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperatures and will therefore melt in the rectum to release the drug. Such materials are cocoa butter and polyethylene glycols.
The compounds of the present invention can also be administered by intranasal, intraocular, intravaginal, and intrarectal routes including suppositories, insufflation, powders and aerosol formulations (for examples of steroid inhalants, see Rohatagi, J. Clin. Pharmacol. 35:1187-1193, 1995; Tjwa, Ann. Allergy Asthma Immunol. 75:107-111, 1995).
The substituted indoles of the present invention can be delivered transdermally, by a topical route, formulated as applicator sticks, solutions, suspensions, emulsions, gels, creams, ointments, pastes, jellies, paints, powders, and aerosols.
Encapsulating materials can also be employed with the compounds of the present invention and the term “composition” can include the active ingredient in combination with an encapsulating material as a formulation, with or without other carriers. For example, the compounds of the present invention can also be delivered as microspheres for slow release in the body. In one embodiment, microspheres can be administered via intradermal injection of drug-containing microspheres, which slowly release subcutaneously (see Rao, J. Biomater Sci. Polym. Ed. 7:623-645, 1995; as biodegradable and injectable gel formulations (see, e.g., Gao, Pharm. Res. 12:857-863, 1995); or, as microspheres for oral administration (see, e.g., Eyles, J. Pharm. Pharmacol. 49:669-674, 1997). Both transdermal and intradermal routes afford constant delivery for weeks or months. Cachets can also be used in the delivery of the compounds of the present invention, e.g., anti-atherosclerotic medicaments.
In another embodiment, the compounds of the present invention can be delivered by the use of liposomes which fuse with the cellular membrane or are endocytosed, i.e., by employing ligands attached to the liposome, or attached directly to the oligonucleotide, that bind to surface membrane protein receptors of the cell resulting in endocytosis. By using liposomes, particularly where the liposome surface carries ligands specific for target cells, or are otherwise preferentially directed to a specific organ, one can focus the delivery of the compound into the target cells in vivo. (See, e.g., Al-Muhammed, J. Microencapsul. 13:293-306, 1996; Chonn, Curr. Opin. Biotechnol. 6:698-708, 1995; Ostro, Am. J. Hosp. Pharm. 46:1576-1587, 1989).
In other cases, the preferred preparation can be a lyophilized powder which may contain, for example, any or all of the following: 1 mM-50 mM histidine, 0.1%-2% sucrose, 2%-7% mannitol, at a pH range of 4.5 to 5.5, that is combined with buffer prior to use.
A pharmaceutical composition of the invention can optionally contain, in addition to a substituted indole, at least one other therapeutic agent useful in the treatment of a disease or condition associated with increased PAI-1 activity.
The pharmaceutical compositions are generally formulated as sterile, substantially isotonic and in full compliance with all Good Manufacturing Practice (GMP) regulations of the U.S. Food and Drug Administration.
E. Determining Dosage Regimens for Substituted Indoles
The present invention provides methods of inhibiting PAI-1 activity in a subject for the treatment of diseases and conditions associated with increased PAI-1 activity using substituted indoles. In an exemplary embodiment of the present invention, a skilled practitioner will treat a subject having a disease associated with elevated PAI-1 levels and/or activity with the compounds of the present invention.
For treatment purposes, the compositions or compounds disclosed herein can be administered to the subject in a single bolus delivery, via continuous delivery (e.g., continuous transdermal, mucosal, or intravenous delivery) over an extended time period, or in a repeated administration protocol (e.g., by an hourly, daily or weekly, repeated administration protocol). The pharmaceutical formulations of the present invention can be administered, for example, one or more times daily, 3 times per week, or weekly. In an exemplary embodiment of the present invention, the pharmaceutical formulations of the present invention are orally administered once or twice daily.
In this context, a therapeutically effective dosage of the biologically active agent(s) can include repeated doses within a prolonged treatment regimen that will yield clinically significant results to alleviate one or more symptoms or detectable conditions associated with increased PAI-1 activity. Determination of effective dosages in this context is typically based on animal model studies followed up by human clinical trials and is guided by determining effective dosages and administration protocols that significantly reduce the occurrence or severity of targeted exposure symptoms or conditions in the subject. Suitable models in this regard include, for example, murine, rat, porcine, feline, non-human primate, and other accepted animal model subjects known in the art. Alternatively, effective dosages can be determined using in vitro models (e.g., immunologic and histopathologic assays). Using such models, only ordinary calculations and adjustments are typically required to determine an appropriate concentration and dose to administer a therapeutically effective amount of the biologically active agent(s) (e.g., amounts that are intranasally effective, transdermally effective, intravenously effective, or intramuscularly effective to elicit a desired response). In alternative embodiments, an “effective amount” or “therapeutically effective dose” of the biologically active agent(s) will simply inhibit or enhance one or more selected biological activity(ies) correlated with a disease or condition, as set forth above, for either therapeutic or diagnostic purposes.
The actual dosage of biologically active agents will of course vary according to factors such as the extent of exposure and particular status of the subject (e.g., the subject's age, size, fitness, extent of symptoms, susceptibility factors, etc), time and route of administration, as well as other drugs or treatments being administered concurrently. Dosage regimens can be adjusted to provide an optimum prophylactic or therapeutic response. By “therapeutically effective dose” herein is meant a dose that produces effects for which it is administered. More specifically, a therapeutically effective dose of the compound(s) of the invention preferably alleviates symptoms, complications, or biochemical indicia of diseases associated with increased PAI-1 activity. The exact dose will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (Vols. 1-3, 1992); Lloyd, 1999, The Art, Science, and Technology of Pharmaceutical Compounding; and Pickar, 1999, Dosage Calculations). A therapeutically effective dose is also one in which any toxic or detrimental side effects of the active agent is outweighed in clinical terms by therapeutically beneficial effects. It is to be further noted that for each particular subject, specific dosage regimens should be evaluated and adjusted over time according to the individual need and professional judgment of the person administering or supervising the administration of the compound.
In an exemplary embodiment of the present invention, unit dosage forms of the compounds are prepared for standard administration regimens. In this way, the composition can be subdivided readily into smaller doses at the physicians direction. For example, unit dosages can 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 can be present in an amount of, for example, from about one gram to about fifteen grams or more, for single or multiple daily administration, according to the particular need of the patient. By initiating the treatment regimen with a minimal daily dose of about one gram, the blood levels of PAI-1 and the patients symptomatic relief analysis can be used to determine whether a larger or smaller dose is indicated. Effective administration of the compounds of this invention can be given at an oral dose of from, for example about 0.1 mg/kg/day to about 1,000 mg/kg/day. Preferably, administration will be from about 10/mg/kg/day to about 600 mg/kg/day, more preferably from about 25 to about 200 mg/kg/day, and even more preferably from about 50 mg/kg/day to about 100 mg/kg/day.
In certain embodiments, the present invention is directed to prodrugs of compounds of formulas 1 and 2. The term “prodrug,” as used herein, means a compound that is convertible in vivo by metabolic means (e.g. by hydrolysis) to a compound of formula 1 or 2. Various forms of prodrugs are known in the art such as those discussed in, for example, Bundgaard, (ed.), Design of Prodrugs, Elsevier (1985); Widder, et al. (ed.), Methods in Enzymology, vol. 4, Academic Press (1985); Krogsgaard-Larsen, et al., (ed). “Design and Application of Prodrugs, Textbook of Drug Design and Development, Chapter 5, 113-191 (1991), Bundgaard, et al., Journal of Drug Delivery Reviews, 8:1-38 (1992), Bundgaard, J. of Pharmaceutical Sciences, 77:285 et seq. (1988); and Higuchi and Stella (eds.) Prodrugs as Novel Drug Delivery Systems, American Chemical Society (1975).
F. Kits
After a pharmaceutical comprising a substituted indole has been formulated in a suitable carrier, it can be placed in an appropriate container and labeled for treatment of a PAI-1 related disorder, e.g., leukemia. Additionally, another pharmaceutical comprising at least one other therapeutic agent useful in the treatment of the PAI-1 related disorder can be placed in the container as well and labeled for treatment of the indicated disease.
Alternatively, a single pharmaceutical comprising a substituted indole and at least one other therapeutic agent useful in the treatment of a PAI-1 related disorder can be placed in an appropriate container and labeled for treatment. For administration of pharmaceuticals comprising substituted indoles and of pharmaceuticals comprising, in a single pharmaceutical, substituted indoles and at least one other therapeutic agent useful in the treatment of a PAI-1 related disorder, such labeling would include, for example, instructions concerning the amount, frequency and method of administration. Similarly, for administration of multiple pharmaceuticals provided in the container, such labeling would include, for example, instructions concerning the amount, frequency and method of administration of each pharmaceutical.
The syntheses of compounds 1-17 are described in examples 1-17 respectively.
Step 1: A large excess of Raney® nickel was added in portions to a stirred solution of 1-(4-nitrophenyl)-1H-indole-2-carboxylic acid (1.40 g, 5.0 mmol), hydrazine (1.0 mL, 32 mmol) in 20 mL of ethanol. After stirring at room temperature for 2 h the catalyst was then removed by filtering through a short pad of Celite®521. The filtrate was concentrated to give 1-(4-aminophenyl)-1H-indole-2-carboxylic acid (1.0 g, 80%) as a beige solid: MS (ESI) m/z 253 (MH+); MS (ESI) m/z 251 ([M-H]−).
Step 2: To a stirred solution of 1-(4-aminophenyl)-1H-indole-2-carboxylic acid (0.4 g, 1.59 mmol) in 10 mL of 1:1 of dichloromethane/sat. aqueous sodium bicarbonate was added 4-trifluoromethoxy-benzenesulfonyl chloride (0.40 g, 1.53 mmol). After being stirred at room temperature for 1 hour the solid was collected and purified by semi-preparative HPLC (Column: Phenomenex C18 Luna 21.6 mm×60 mm, 5 μM; Solvent A: Water (0.1% TFA buffer); Solvent B: Acetonitrile (0.1% TFA buffer); Solvent Gradient: Time 0:0% B; 10 min: 100% B; Hold 100% B 5 min. Flow Rate: 22.5 mL/min) to afford 0.105 g (15%) of the title compound as a pale yellow solid: 1H NMR (DMSO-d6) δ 6.89 (d, J=8.3 Hz, 1H), 7.10-7.30 (m, 6H), 7.39 (d, J=2.1 Hz, 1H), 7.60 (d, J=7.2 Hz, 2H), 7.71 (d, J=6.0 Hz, 1H), 7.90-8.00 (m, 2H), 10.63 (br s, 1H), 12.74 (br s, 1H); MS (ESI) m/z 477 (MH+); MS (ESI) m/z 475 ([M-H]−); HRMS calcd for C22H16F3N2O5S: 477.0721; found (ESI+): 477.0719; Anal. calcd for C22H15F3N2O5S.0.2H2O: C, 55.05; H, 3.23; N, 5.84. Found: C, 54.97; H, 3.41; N, 5.82.
A solution of 1-(4-aminophenyl)-1H-indole-2-carboxylic acid (0.4 g, 1.59 mmol) and 2,5-dimethoxytetrahydrofuran (0.50 g, 3.78 mmol) in 15 mL of glacial acetic acid was heated at 90° C. for 1.5 hours. After removing the solvent the residue was purified by semi-preparative HPLC to afford the title compound (0.095 mg, 20%) as a beige solid: 1H NMR (DMSO-d6) δ 6.32 (s, 2H), 7.08 (d, J=8.4 Hz, 1H), 7.15-7.25 (m, 1H), 7.25-7.35 (m, 1H), 7.40-7.55 (m, 5H), 7.60-7.70 (m, 3H), 12.82 (br s, 1H); MS (ESI) m/z 303 (MH+); MS (ESI) m/z 301 ([M-H]−); HRMS calcd for C19H15N2O2: 303.1122; found (ESI+): 303.1124; Anal. calcd for C19H14N2O2 0.1H2O: C, 75.0; H, 4.71; N, 9.21. Found: C, 74.90; H, 4.42; N, 9.33.
Step 1: A mixture of 4-nitrobenzyl bromide (1.85 g, 8.56 mmol), 1H-indole-2-carboxylic acid ethyl ester (1.62 g, 8.56 mmol) and potassium carbonate (4.17 g, 30 mmol) in 100 mL of acetone was refluxed for 1 day. After cooling to room temperature the reaction was quenched by addition of water and then partitioned between water and methylene chloride. The organic phase was dried over MgSO4, and concentrated to yield ethyl 1-(4-nitrobenzyl)-1H-indole-2-carboxylate as an off-white solid (2.0 g, 72%): 1H NMR (DMSO-d6) δ 1.27 (t, J=7.1 Hz, 3H), 4.26 (q, J=7.1 Hz, 3H), 5.99 (s, 2H), 7.18 (t, J=7.0 Hz, 1H), 7.22 (d, J=9.0 Hz, 2H), 7.34 (t, J=7.0 Hz, 1H), 7.43 (s, 1H), 7.58 (d, J=13.8 Hz, 1H), 7.76 (d, J=13.8 Hz, 1H), 8.15 (d, J=9.0 Hz, 2H); MS (ESI) m/z 325 (MH+).
Step 2: A large excess of Raney® nickel was added in portions to a stirred solution of ethyl 1-(4-nitrobenzyl)-1H-indole-2-carboxylate (0.65 g, 2 mmol), hydrazine (0.5 mL, 16 mmol) in 25 mL of ethanol. After stirring at room temperature for 2 hours the catalyst was then removed by filtering through a short pad of Celite®521. The filtrate was concentrated to give ethyl 1-(4-aminobenzyl)-1H-indole-2-carboxylate (MS (ESI) m/z 294.2 (MH+) as a gummy solid. The gummy solid was then dissolved in 15 mL of methylene chloride. To the solution was added diisopropylethylamine (0.5 mL, 2.9 mmol) and biphenyl 4-sulfonyl chloride (0.5 g, 1.0 mmol). After being stirred at room temperature overnight the reaction was concentracted and the residue was re-dissolved in 15 mL of 2:1:1 THF/MeOH/water. Lithium hydroxide monohydrate (0.2 g, 4.8 mmol) was added and the mixture was stirred at room temperature overnight. Most of the organic solvents was removed and the reaction mixture was made acidic (pH 6) with glacial acetic acid, and the solid was collected and purified by semi-preparative HPLC. The product was collected based on UV absorption and concentrated to give the title compound as a brown solid (0.11 g, 22%): 1H NMR (DMSO-d6) δ 5.75 (s, 2H), 6.93 (d, J=8.9 Hz, 2H), 7.01 (d, J=8.9 Hz, 2H), 7.09 (t, J=8.0 Hz, 1H), 7.43 (t, J=8.0 Hz, 1H), 7.49 (s, 1H), 7.40-7.55 (m, 4H); 7.65-7.80 (m, 3H), 7.75-7.82 (m, 4H), 10.30 (s, 1H), 12.93 (s, 1H); HRMS calcd for C28H23N2O4S: 483.1367; found (ESI+): 483.1374; Anal. calcd for C28H22N2O4S 0.2H2O: C, 69.18; H, 4.64; N, 5.76. Found: C, 69.21; H, 4.60; N, 5.78.
Step 1: 1-(4-Nitro-benzyl)-1H-indole-5-carboxylic acid methyl ester was prepared from 1H-indole-5-carboxylic acid methyl ester and 4-nitrobenzyl bromide followed the procedure of Example 3 Step 1 as a yellow powder: 1H NMR (DMSO-d6) δ 3.84 (s, 3H), 5.67 (s, 2H), 6.71 (dd, J=3.2, 0.8 Hz, 1H), 7.40 (d, J=8.8 Hz, 2H), 7.54 (d, J=8.7 Hz, 1H), 7.67 (d, J=0.8 Hz, 1H), 7.73 (dd, J=8.8, 1.7 Hz, 1H), 8.19 (d, J=8.8 Hz, 2H), 8.29 (d, J=1.0 Hz, 1H); MS (ESI) m/z 311 (MH+), 309 ([M-H]−); HRMS calcd for C17H15N2O4: 311.1030; found (ESI+): 311.1021.
Step 2: The title compound was prepared from 1-(4-nitro-benzyl)-1H-indole-5-carboxylic-acid methyl ester and 4-tert-butylbenzene sulfonyl chloride followed the procedure of Example 3 Step 2 as a light brown solid: 1H NMR (DMSO-d6) δ 1.24 (s, 9H), 5.33 (s, 2H), 6.59 (d, J=3.2 Hz, 1H), 7.02 (d, J=8.7 Hz, 2H), 7.07 (d, J=8.7 Hz, 2H), 7.45 (d, J=8.7 Hz, 1H), 7.51-7.54 (m, 3H), 7.65 (d, J=8.5 Hz, 2H), 7.69 (d, J=1.7 Hz, 1H), 8.21 (d, J=1.3 Hz, 1H), 10.23 (br s, 1H), 12.39 (br s, 1H); MS (ESI) m/z 463 (MH+), 461 ([M-H]−); HRMS calcd for C26H26N2O4S: 463.1690; found (ESI+): 463.1681.
The title compound was prepared from 1-(4-nitro-benzyl)-1H-indole-5-carboxylic acid methyl ester and bisphenyl-4-sulfonyl chloride followed the procedure of Example 3 Step 2 as a white solid: 1H NMR (DMSO-d6) δ 5.34 (s, 3H), 6.59 (d, J=3.1 Hz, 1H), 7.06 (d, J=11.2 Hz, 2H), 7.10 (d, J=8.7 Hz, 2H), 7.40-7.48 (m, 4H), 7.52 (d, J=3.1 Hz, 1H), 7.67-7.70 (m, 3H), 7.75-7.85 (m, 4H), 8.21 (d, J=1.4 Hz, 1H), 10.34 (br s, 1H), 12.37 (br s, 1H); MS (ESI) m/z 483 (MH+), 481 ([M-H]−); HRMS calcd for C28H23N2O4S: 483.1376; found (ESI+): 483.1368.
Step 1: 1-(4-Nitro-benzyl)-1H-indole-3-carboxylic acid methyl ester (113220-11) was prepared from 1H-indole-3-carboxylic acid methyl ester and 4-nitrobenzyl bromide followed the procedure of Example 3 Step 1 as a yellow powder: 1H NMR (DMSO-d6) δ 3.83 (s, 3H), 5.70 (s, 2H), 7.23 (ddd, J=9.9, 4.6, 0.8 Hz, 2H), 7.47 (d, J=8.8 Hz, 2H), 7.50 (ddd, J=9.9, 4.6, 0.8 Hz, 1H), 8.05 (ddd, J=9.9, 4.6, 0.8 Hz, 1H), 8.19 (d, J=8.8 Hz, 2H), 8.40 (s, 1H); MS (ESI) m/z 311 (MH+), 309 ([M-H]−).
Step 2: The title compound was prepared from 1-(4-nitro-benzyl)-1H-indole-3-carboxylic acid methyl ester and 4-tert-butylbenzenesulfonyl chloride followed the procedure of Example 3 Step 2 as a light brown solid: 1H NMR (DMSO-d6) δ 1.24 (s, 9H), 5.36 (s, 2H), 7.04 (d, J=8.5 Hz, 2H), 7.13-7.18 (m, 4H), 7.45 (dd, J=6.1, 2.6 Hz, 1H), 7.52 (d, J=8.8 Hz, 2H), 7.66 (d, J=8.8 Hz, 2H), 8.00 (dd, J=6.1, 2.6 Hz, 1H), 8.13 (s, 1H), 10.23 (br s, 1H), 11.98 (br s, 1H); MS (ESI) m/z 463 (MH+), 461 ([M-H]−); HRMS calcd for C26H27N2O4S: 463.1690; found (ESI+): 463.1681; Anal. calcd for C26H26N2O4S.0.06 TFA: C, 67.51; H, 5.67; N, 6.06. Found: C, 67.40; H, 5.65; N, 6.04.
The title compound was prepared from 1-(4-nitro-benzyl)-1H-indole-3-carboxylic acid methyl ester and bisphenyl-4-sulfonyl chloride followed the procedure of Example 3 Step 2 as an off-white solid: 1H NMR (DMSO-d6) δ 5.37 (s, 2H), 7.07 (d, J=8.7 Hz, 2H), 7.13-7.18 (m, 4H), 7.40-7.51 (m, 4H), 7.67 (d, J=7.1 Hz, 2H), 7.75-7.85 (m, 4H), 8.00 (dd, J=5.8, 0.7 Hz, 1H), 8.14 (s, 1H), 10.36 (br s, 1H), 12.01 (br s, 1H); MS (ESI) m/z 483 (MH+), 481 ([M-H]−); HRMS calcd for C28H23N2O4S, 483.1376; found (ESI+): 483.1367; Anal. calcd for C28H22N2O4S.0.1 TFA: C, 68.57; H, 4.51; N, 5.67. Found: C, 68.67; H, 4.50; N, 5.70.
Step 1: 1-(4-Nitro-benzyl)-1H-indole-4-carboxylic acid methyl ester was prepared from 1H-indole-4-carboxylic acid methyl ester and 4-nitrobenzyl bromide followed the procedure of Example 3 Step 1 as a yellow powder: 1H NMR (DMSO-d6) δ 3.90 (s, 3H), 5.69 (s, 2H), 7.05 (dd, J=3.1, 0.9 Hz, 1H), 7.22 (t, J=7.6 Hz, 1H), 7.36 (d, J=8.9 Hz, 2H), 7.75 (td, J=7.6, 3.1 Hz, 3H), 8.18 (d, J=8.9 Hz, 2H); MS (ESI) m/z 311 (MH+), 309 ([M-H]−); Anal. calcd for C17H14N2O4: C, 65.80; H, 4.55; N, 9.03. Found: C, 65.81; H, 4.50; N, 8.98.
Step 2: The title compound was prepared from 1-(4-nitro-benzyl)-1H-indole-4-carboxylic acid methyl ester and 4-tert-butylbenzenesulfonyl chloride followed the procedure of Example 3 Step 2 as a pale pink solid: 1H NMR (DMSO-d6) δ 1.24 (s, 9H), 5.35 (s, 2H), 6.96 (d, J=3.1 Hz, 1H), 7.03 (d, J=8.7 Hz, 2H), 7.07 (d, J=8.0 Hz, 2H), 7.15 (t, J=7.8 Hz, 1H), 7.53 (d, J=8.8 Hz, 2H), 7.57 (d, J=3.2 Hz, 1H), 7.62-7.68 (m, 3H), 7.70 (dd, J=7.5, 0.9 Hz, 1H), 10.26 (br s, 1H), 12.59 (br s, 1H); MS (ESI) m/z 463 (MH+), 461 ([M-H])−); HRMS calcd for C26H27N2O4S: 463.1690; found (ESI+): 463.1674.
The title compound was prepared from 1-(4-nitro-benzyl)-1H-indole-4-carboxylic acid methyl ester and bisphenyl-4-sulfonyl chloride followed the procedure of Example 3 Step 2 as an off-white solid: 1H NMR (DMSO-d6) δ 5.36 (s, 2H), 6.96 (d, J=3.1 Hz, 1H), 7.05 (d, J=8.0 Hz, 2H), 7.09 (d, J=9.0 Hz, 2H), 7.15 (t, J=7.8 Hz, 1H), 7.40-7.47 (m, 1H), 7.49-7.51 (m, 2H), 7.57 (d, J=3.1 Hz, 1H), 7.64-7.71 (m, 4H), 7.75-7.85 (m, 4H), 10.33 (br s, 1H), 12.58 (br s, 1H); MS (ESI) m/z 483 (MH+); 481 ([M-H]−); HRMS calcd for C28H23N2O4S: 483.1376; found (ESI+): 483.1367.
Step 1: 1-(4-Nitrobenzyl)-1H-indole-6-carboxylic acid methyl ester was prepared from 1H-indole-6-carboxylic acid methyl ester and 4-nitrobenzyl bromide followed the procedure of Example 3 Step 1 as a yellow powder: 1H NMR (DMSO-d6) δ 3.85 (s, 3H), 5.75 (s, 2H), 6.66 (dd, J=3.1, 0.6 Hz, 1H), 7.34 (d, J=8.7 Hz, 2H), 7.60 (t, J=2.8 Hz, 1H), 7.79 (d, J=3.1 Hz, 1H), 8.07 (s, 1H), 8.15 (s, 1H), 8.19 (d, J=8.8 Hz, 1H), 8.28 (d, J=8.7 Hz, 2H); MS (ESI) m/z 311 (MH+); 309 ([M-H]−).
Step 2: The title compound was prepared from 1-(4-nitro-benzyl)-1H-indole-6-carboxylic acid methyl ester and bisphenyl-4-sulfonyl chloride followed the procedure of Example 3 Step 2 as a light yellow solid: 1H NMR (DMSO-d6) δ 5.40 (s, 2H), 6.55 (d, J=3.9 Hz, 1H), 7.06 (s, 4H), 7.40-7.43 (m, 1H), 7.47-7.51 (m, 2H), 7.60 (d, J=1.2 Hz, 1H), 7.63 (d, J=3.1 Hz, 1H), 7.67 (dt, J=7.0, 1.2 Hz, 2H), 7.75-7.85 (m, 4H), 7.99 (d, J=0.9 Hz, 1H), 10.32 (br s, 1H), 12.55 (br s, 1H); MS (ESI) m/z 483 (MH+); 481 ([M-H]−); HRMS calcd for C28H23N2O4S: 483.1376; found (ESI+): 483.1368; Anal. calcd for C28H22N2O4S.0.02 TFA C, 69.21; H, 4.60; N, 5.76. Found: C, 69.01 H, 4.48 N, 5.78.
Step 1: A solution of N-bromosuccinimide (4.94 g, 27.7 mmol) in DMF (20 mL) was added dropwise to a stirred solution of ethyl 1H-indole-2-carboxylate (5.0 g, 26.4 mmol) in DMF (15 mL) at 0˜5° C. After the addition was completed, the reaction was warmed to room temperature and stirring was continued for an additional 1.5 h. The reaction mixture was poured over ice water and the resulting precipitate was collected and dried to give 3-bromo-1H-indole carboxylic acid ethyl ester (6.68 g, 81%) as a white solid: 1H NMR (DMSO-d6) δ 1.37 (t, J=7.2 Hz, 3H), 4.38 (q, J=7.2 Hz, 2H), 7.18-7.22 (m, 1H), 7.34-7.38 (m, 1H), 7.50 (dd, J=8.3, 0.8 Hz, 1H), 7.55 (dd, J=8.3, 0.8 Hz, 1H), 12.22 (br s, 1H); MS (ESI) m/z 268 (MH+); 266 ([M-H]−).
Step 2: A mixture of 3-bromo-1H-indolecarboxylic acid ethyl ester (0.50 g, 1.87 mmol), phenylboronic acid (0.27 g, 2.25 mmol), 2 M aqueous sodium carbonate (8 mL), tetrakis(triphenylphosphine)palladium (0) (0.20 g, 0.17 mmol) in ethanol (16 mL) and toluene (30 ml) was heated at 65° C. for 16 h and then cooled. The reaction mixture was diluted with 1 N hydrochloric acid and then extracted with ethyl acetate. The organic extracts were washed with water, dried over magnesium sulfate and concentrated. Flash silica gel chromatography using ethyl acetate/hexane as elutant gave 0.34 g (68%) of 3-phenyl-1H-indolecarboxylic acid ethyl ester as a pink solid: 1H NMR (DMSO-d6) δ 1.18 (t, J=7.2 Hz, 3H), 4.22 (q, J=7.2 Hz, 2H), 7.07-7.11 (m, 1H), 7.29-7.33 (m, 1H) 7.45 (dd, J=8.3, 0.8 Hz, 1H), 7.48-7.50 (m, 5H), 7.50 (dd, J=8.3, 0.8 Hz, 1H), 11.90 (br s, 1H); MS (ESI) m/z 266 (MH+); 264 ([M-H]−).
Step 3: 1-(4-Nitro-benzyl)-3-phenyl-1H-indolecarboxylic acid ethyl ester was prepared from of 3-phenyl-1H-indolecarboxylic acid ethyl ester and 4-nitrobenzyl bromide followed the procedure of Example 3 Step 1 as a yellow powder: 1H NMR (DMSO-d6) δ 1H NMR (DMSO-d6) δ 0.91 (t, J=7.2 Hz, 3H), 4.05 (q, J=7.2 Hz, 2H), 5.97 (s, 2H), 7.17-7.23 (m, 1H), 7.32 (d, J=8.8 Hz, 2H), 7.36-7.42 (m, 1H), 7.38-7.42 (m, 1H), 7.45-7.48 (m, 4H), 7.52 (d, J=8.2 Hz, 1H), 7.62 (d, J=8.2 Hz, 1H), 8.19 (d, J=8.8 Hz, 2H); MS (ESI) m/z 401 (MH+); 399 ([M-H]−); HRMS calcd for C24H21N2O4: 401.1490; found (ESI+): 401.1494; Anal. calcd for C24H20N2O4.0.05 TFA C, 71.99; H, 5.03; N, 7.00 Found: C, 71.27; H, 4.98; N, 6.90.
Step 4: 1-(4-Amino-benzyl)-3-phenyl-1H-indole indolecarboxylic acid ethyl ester was prepared from 1-(4-nitro-benzyl)-3-phenyl-1H-indolecarboxylic acid ethyl ester followed the procedure of Example 1 Step 1 as a yellow syrup: 1H NMR (DMSO-d6) δ 0.98 (t, J=7.2 Hz, 3H), 4.11 (q, J=7.2 Hz, 2H), 4.99 (br s, 2H), 5.59 (s, 2H), 6.44 (d, J=8.41 Hz, 2H), 6.85 (d, J=8.4 Hz, 2H), 7.13 (td, J=7.1, 0.8 Hz, 1H) 7.34 (td, J=7.1, 0.8 Hz, 1H), 7.38-7.41 (m, 3H), 7.44-7.49 (m, 3H), 7.69 (d, J=8.4 Hz, 1H); MS (ESI) m/z 371 (MH+); 369 ([M-H]−); Anal. calcd for C24H22N2O2.0.04 TFA C, 71.12; H, 5.92; N, 7.47. Found: C, 71.03; H, 5.87; N, 7.36.
Step 5: The title compound was prepared from 1-(4-amino-benzyl)-3-phenyl-1H-indole indolecarboxylic acid ethyl ester and benzenesulfonyl chloride followed the procedure of Example 3 Step 2 as an off-white solid: 1H NMR (DMSO-d6) δ 5.72 (s, 2H), 6.95-7.00 (m, 4H), 7.09-7.13 (m, 1H), 7.27-7.31 (m, 1H), 7.33-7.37 (m, 1H), 7.42-7.45 (m, 5H), 7.49-7.55 (m, 3H), 7.57-7.61 (m, 1H), 7.72 (dd, J=7.1, 1.4 Hz, 2H), 10.26 (br s, 1H), 12.83 (br s, 1H); MS (ESI) m/z 481 ([M-H]−); HRMS calcd for C28H21N2O4S: 481.1224; found (ESI−): 481.1228.
The title compound was prepared from 1-(4-nitro-benzyl)-3-phenyl-1H-indole indolecarboxylic acid ethyl ester and methanesulfonyl chloride followed the procedure of Example 3 Step 2 as an off-white solid: 1H NMR (DMSO-d6) δ 2.94 (s, 3H), 5.80 (s, 2H), 7.05-7.15 (m, 5H), 7.31-7.38 (m, 2H), 7.43-7.47 (m, 5H), 7.62 (d, J=8.4 Hz, 1H), 9.68 (br s, 1H), 12.87 (br s, 1H); MS (ESI) m/z 419 ([M-H]−); HRMS calcd for C23H19N2O4S: 419.1068; found (ESI−): 419.1074.
Step 1: 3-Thiophene-2-yl-1H-indole-2-carboxylic acid ethyl ester was prepared from 3-bromo-1H-indolecarboxylic acid ethyl ester and 2-thiopheneboronic acid followed the procedure of Example 11 Step 2 as a light yellow powder: 1H NMR (DMSO-d6) δ 1.27 (t, J=7.2 Hz, 3H), 4.30 (q, J=7.2 Hz, 2H), 7.12-7.16 (m, 1H), 7.17-7.20 (m, 1H), 7.31-7.34 (m, 1H), 7.36 (dd, J=5.1, 1.5 Hz, 1H), 7.51 (d, J=8.3 Hz, 1H), 7.63 (d, J=6.3 Hz, 1H), 7.71 (d, J=8.2 Hz, 1H), 12.02 (br s, 1H); MS (ESI) m/z 272 (MH+); 270 ([M-H]−).
Step 2: 1-(4-Nitro-benzyl)-3-thiophene-2-yl-1H-indole-2-carboxylic acid ethyl ester was prepared from 3-thiophene-2-yl-1H-indole-2-carboxylic acid ethyl ester and 4-nitrobenzyl bromide followed the procedure of Example 3 Step 1 as a yellow powder: 1H NMR (DMSO-d6) δ 1.02 (t, J=7.2 Hz, 3H), 4.15 (q, J=7.2 Hz, 2H), 7.20 (dd, J=6.9, 3.4 Hz, 1H), 7.24 (dd, J=6.2, 0.8 Hz, 1H), 7.25-7.27 (m, 1H), 7.31 (d, J=8.8 Hz, 2H), 7.37-7.43 (m, 1H), 7.62 (d, J=8.4 Hz, 1H), 7.68 (dd, J=5.2, 1.2 Hz, 1H), 7.71 (d, J=4.1 Hz, 1H), 8.19 (d, J=8.8 Hz, 2H); MS (ESI) m/z 407 (MH+); 405 ([M-H]−); Anal. calcd for C22H18N2O4S: C, 65.01; H, 4.46; N, 6.89 Found: C, 65.04; H, 6.62; N, 6.82.
Step 3: 1-(4-Amino-benzyl)-3-thiophene-2-yl-1H-indole-2-carboxylic acid ethyl ester was prepared from 1-(4-nitro-benzyl)-3-thiophene-2-yl-1H-indole-2-carboxylic acid ethyl ester followed the procedure of Example 1 Step 1: 1H NMR (DMSO-d6) δ 1.10 (t, J=7.2 Hz, 3H), 4.20 (q, J=7.2 Hz, 2H), 5.01 (br s, 2H), 5.55 (s, 2H), 6.45 (d, J=8.5 Hz, 2H), 6.84 (d, J=8.5 Hz, 2H), 7.16-7.20 (m, 3H), 7.35 (td, J=7.3, 1.0 Hz, 1H), 7.62 (dd, J=5.2, 1.2 Hz, 1H), 7.65 (d, J=4.1 Hz, 1H), 7.71 (d, J=8.8 Hz, 1H); MS (ESI) m/z 377 (MH+); 375 ([M-H]−); Anal. calcd for C22H20N2O2S.0.05 TFA C, 69.46; H, 5.29; N, 7.33. Found: C, 69.66; H, 5.04; N, 7.72.
Step 4: The title compound was prepared from 1-(4-amino-benzyl)-3-thiophene-2-yl-1H-indole-2-carboxylic acid ethyl ester and benzenesulfonyl chloride followed the procedure of Example 3 Step 2 as a light brown solid: 1H NMR (DMSO-d6) δ 5.67 (s, 2H), 6.95-7.05 (m, 4H), 7.14-7.23 (m, 3H), 7.30-7.35 (m, 1H), 7.49-7.75 (m, 8H), 10.26 (br s, 1H), 13.24 (br s, 1H); MS (ESI) m/z 489 (MH+); HRMS calcd for C26H21N2O4S2: 489.0940; found (ESI+): 489.0935.
The title compound was prepared from 1-(4-nitro-benzyl)-3-thiophene-2-yl-1H-indole-2-carboxylic acid ethyl ester and methanesulfonyl chloride followed the procedure of Example 3 Step 2 as a light yellow solid: 1H NMR (DMSO-d6) δ 2.94 (s, 3H), 5.76 (s, 2H), 7.08-7.24 (m, 7H), 7.34 (td, J=8.4, 1.2 Hz, 1H), 7.60-7.70 (m, 3H), 9.69 (br s, 1H), 13.25 (br s, 1H); MS (ESI) m/z 427 (MH+); MS (ESI) m/z 425 ([M-H]−); HRMS calcd for C21H19N2O4S2: 427.0784; found (ESI+): 427.0779.
A mixture of 1-(4-amino-benzyl)-3-phenyl-1H-indolecarboxylic acid ethyl ester (0.20 g, 0.54 mmol), acetonylacetone (0.16 mL, 1.35 mmol), and toluene (15 mL) was heated at reflux under nitrogen using a Dean-Stark trap for 6 h. The reaction was cooled and concentrated. The residue was re-dissolved in 10 mL of 2:1:1 THF/MeOH/water. Lithium hydroxide monohydrate (0.09 g, 2.1 mmol) was added and the mixture was stirred at room temperature overnight. Most of the organic solvents was removed and the reaction mixture was made acidic (pH 6) with glacial acetic acid, and the solid was collected and purified by HPLC to afford the title compound (0.027 g, 6%) as a brown solid: 1H NMR (DMSO-d6) δ 1.91 (s, 6H), 5.75 (s, 2H), 7.01-7.30 (m, 6H), 7.32-7.38 (m, 2H), 7.40-7.50 (m, 6H), 7.68 (d, J=8.54 Hz, 1H), 12.80 (br s, 1H); MS (ESI) m/z 421 (MH+); MS (ESI) m/z 419 ([M-H]−); HRMS calcd for C28H26N2O2: 421.1905; found (ESI+): 421.1915.
Phenylboronic acid (0.16 g, 0.81 mmol), Cu(OAc)2 (0.049 g, 0.27 mmol), and myristic acid (0.037 g, 0.16 mmol) were combined in a 100-mL round-bottom flask with a large stir bar. A rubber septum was attached, and dry toluene (2 mL), 2,6-lutidine (0.066 mL, 0.57 mmol), and 1-(4-amino-benzyl)-3-phenyl-1H-indole indolecarboxylic acid ethyl ester (0.20 g, 0.54 mmol) were successively added. The resulting mixture was stirred at a high rate for 24 h, diluted with ethyl acetate (10 mL), filtered through a plug of silica gel, and then purified by column chromatography (5 to 25% EtOAc/hexanes) to give a gummy solid. To a stirred solution of the gummy solid in 15 mL of 2:1:1 THF/MeOH/water was added lithium hydroxide monohydrate (0.1 g, 2.4 mmol). The reaction was stirred at 40° C. for 5 h. Most of the organic solvents was removed and the reaction mixture was made acidic (pH 6) with glacial acetic acid, and the solid was collected and purified by HPLC as a white powder (0.023 g, 10%): 1H NMR (DMSO-d6) δ 5.64 (s, 2H), 6.77 (t, J=7.4 Hz, 1H), 6.96 (d, J=8.5 Hz, 2H), 7.00 (dd, J=7.6, 0.9 Hz, 2H), 7.04 (t, J=7.1 Hz, 1H), 7.15-7.19 (m, 4H), 7.25 (t, J=7.11 Hz, 1H), 7.39 (t, J=7.4 Hz, 2H), 7.46-7.53 (m, 1H), 7.53 (t, J=7.4 Hz, 2H), 7.54-7.56 (m, 2H), 8.08 (br s, 1H), 13.01 (br s, 1H); MS (ESI) m/z 419 (MH+); MS (ESI) m/z 417 ([M-H]−); HRMS calcd for C28H23N2O2: 419.1757; found (ESI+): 419.1755.
The title compound was prepared from 1-(4-amino-benzyl)-3-thiophene-2-yl-1H-indole-2-carboxylic acid ethyl ester and phenylboronic acid followed the procedure of Example 16 as a light green solid: 1H NMR (DMSO-d6) δ 5.70 (s, 2H), 6.78 (td, J=7.4, 1.0 Hz, 1H), 6.98 (d, J=8.7 Hz, 2H), 7.01 (dd, J=8.5, 1.0 Hz, 2H), 7.04 (d, J=8.7 Hz, 2H), 7.16 (t, J=3.5 Hz, 1H), 7.17 (dd, J=5.2, 1.6 Hz, 2H), 7.21 (dd, J=6.9, 2.1 Hz, 2H), 7.33-7.37 (m, 1H), 7.61 (dd, J=5.1, 1.3 Hz, 1H), 7.66 (t, J=8.7 Hz, 2H), 8.12 (br s, 1H), 13.34 (br s, 1H); MS (ESI) m/z 425 (MH+); MS (ESI) m/z 423 ([M-H]−); HRMS calcd for C26H21N2O2S: 425.1321; found (ESI+): 425.1319.
Test compounds are dissolved in DMSO at a final concentration of 10 mM, then diluted 100× in physiologic buffer. The inhibitory assay is initiated by the addition of the test compound (1-100 μM final concentration, maximum DMSO concentration of 0.2%) in a pH 6.6 buffer containing 140 nM recombinant human plasminogen activator inhibitor-1 (PAI-1; Molecular Innovations, Royal Oak, Mich.). Following a 1 hour incubation at room temperature, 70 nM of recombinant human tissue plasminogen activator (tPA) is added, and the combination of the test compound, PAI-1 and tPA is incubated for an additional 30 minutes. Following the second incubation, Spectrozyme-tPA (American Diagnostica, Greenwich, Conn.), a chromogenic substrate for tPA, is added and absorbance read at 405 nm at 0 and 60 minutes. Relative PAI-1 inhibition is equal to the residual tPA activity in the presence of the test compound and PAI-1. Control treatments include the complete inhibition of tPA by PAI-1 at the molar ratio employed (2:1), and the absence of any effect of the test compound on tPA alone.
This assay is based upon the non-SDS dissociable interaction between tPA and active PAI-1. Assay plates are initially coated with human tPA (10 μg/ml). Test compounds of the present invention are dissolved in DMSO at 10 mM, then diluted with physiologic buffer (pH 7.5) to a final concentration of 1-50 μM. Test compounds are incubated with human PAI-1 (50 ng/ml) for 15 minutes at room temperature. The tPA-coated plate is washed with a solution of 0.05% Tween 20 and 0.1% BSA, then the plate is blocked with a solution of 3% BSA. An aliquot of the substituted indole/PAI-1 solution is then added to the tPA-coated plate, incubated at room temperature for 1 hour, and washed. Active PAI-1 bound to the plate is assessed by adding an aliquot of a 1:1000 dilution of the 33B8 monoclonal antibody against human PAI-1, and incubating the plate at room temperature for 1 hour (Molecular Innovations, Royal Oak, Mich.). The plate is again washed, and a solution of goat anti-mouse IgG-alkaline phosphatase conjugate is added at a 1:50,000 dilution in goat serum. The plate is incubated 30 minutes at room temperature, washed, and a solution of alkaline phosphatase substrate is added. The plate is incubated 45 minutes at room temperature, and color development is determined at OD405nm. The quantitation of active PAI-1 bound to tPA at varying concentrations of the test compound is used to determine the IC50. Results are analyzed using a logarithmic best-fit equation. The assay sensitivity is 5 ng/ml of human PAI-1 as determined from a standard curve ranging from 0-100 ng/ml.
The compounds of the present invention inhibited Plasminogen Activator Inhibitor-1 summarized in Table 1.
Although the foregoing invention has been described in detail by way of example for purposes of clarity of understanding, it will be apparent to the artisan that certain changes and modifications are comprehended by the disclosure and can be practiced without undue experimentation within the scope of the appended claims, which are presented by way of illustration not limitation.
All publications and patent documents cited above are hereby incorporated by reference in their entirety for all purposes to the same extent as if each were so individually denoted.
This application is a continuation of U.S. application Ser. No. 10/947,726 filed Sep. 23, 2004, which claims the benefit of U.S. Provisional Application No. 60/505,820 filed Sep. 25, 2003, each of which is incorporated herein by reference.
| Number | Date | Country | |
|---|---|---|---|
| 60505820 | Sep 2003 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | 10947726 | Sep 2004 | US |
| Child | 11881513 | Jul 2007 | US |
