WO2005082340A2 - Modulation of inflammatory and metastatic processes - Google Patents

Modulation of inflammatory and metastatic processes Download PDF

Info

Publication number
WO2005082340A2
WO2005082340A2 PCT/US2005/005316 US2005005316W WO2005082340A2 WO 2005082340 A2 WO2005082340 A2 WO 2005082340A2 US 2005005316 W US2005005316 W US 2005005316W WO 2005082340 A2 WO2005082340 A2 WO 2005082340A2
Authority
WO
WIPO (PCT)
Prior art keywords
groups
substituted
unsubstituted
alkyl
aryl
Prior art date
Application number
PCT/US2005/005316
Other languages
French (fr)
Other versions
WO2005082340A3 (en
Inventor
Sang H. Lee
Carla C. Heise
Original Assignee
Chiron Corporation
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Chiron Corporation filed Critical Chiron Corporation
Priority to RU2006133536/15A priority Critical patent/RU2377988C2/en
Priority to CA2556872A priority patent/CA2556872C/en
Priority to BRPI0507891-1A priority patent/BRPI0507891A/en
Priority to JP2006554253A priority patent/JP5019884B2/en
Priority to AU2005216904A priority patent/AU2005216904B2/en
Priority to CN2005800095231A priority patent/CN1960731B/en
Priority to EP05723338A priority patent/EP1718306A2/en
Publication of WO2005082340A2 publication Critical patent/WO2005082340A2/en
Publication of WO2005082340A3 publication Critical patent/WO2005082340A3/en
Priority to IL177574A priority patent/IL177574A0/en

Links

Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/395Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/4965Non-condensed pyrazines
    • A61K31/497Non-condensed pyrazines containing further heterocyclic rings
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/395Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47Quinolines; Isoquinolines
    • A61K31/4709Non-condensed quinolines and containing further heterocyclic rings
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/395Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/53751,4-Oxazines, e.g. morpholine
    • A61K31/53771,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00Antineoplastic agents
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00Antineoplastic agents
    • A61P35/04Antineoplastic agents specific for metastasis
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

  • the present invention provides methods for using compounds to modulate inflammatory responses and processes related to tumor metastasis.
  • the invention further provides methods for monitoring the effects of the compounds of the invention by measuring the levels of ICAM, VCAM, or E-selectin molecules in a subject treated with the compounds.
  • Amino quinolinone benzimidazolyl compounds such as 4-amino-5- fluoro-3 - [6-(4-methylpiperazin- 1 -yl)- 1 H-benzimidazol-2-yl] quinolin-2( 1 H)-one and their tautomers and salts are potent inhibitors of various class kinases such as VEGFR2 (KDR, Flk-1), FGFR1 and PDGFR ⁇ with IC 50 s ranging from 10-27 nM. See U.S. Patent No. 6,605,617, U.S. Patent Application No. 10/644,055, and U.S. Patent Application No.
  • NCAM vascular cell adhesion molecule
  • VCAM vascular cell adhesion molecule
  • ICAM inducible cell adhesion molecule
  • ICAM is also expressed in endothelial cells and various cells including fibroblasts, hematopoietic cells, and tumor cells. The soluble form of ICAM present in the plasma is generated by proteolytic cleavage of membrane-associated molecules.
  • E-Selectin endofhelial leukocyte adhesion molecule
  • E-selectin endofhelial leukocyte adhesion molecule
  • a high concentration of soluble ICAM, VCAM, and E-selectin is considered a marker of endothelial cell activation during tumor development, metastasis, and inflammatory responses.
  • These cell adhesion molecules localized on endothelial cells can mediate adhesion of metastatic tumor cells and allow extravasation into the vessels. It is of interest that these molecules are inducible, being poorly expressed on normal endothelial cells but capable of being expressed highly after exposure to cytokines such as IL-1 or TNF-a. In addition, some of these molecules are preferentially expressed in different vascular beds, with VCAM being abundant in the lung and E-selectin in the liver.
  • MMPs Matrix metalloproteases
  • ECM extracellular matrix
  • MMP-9/gelatinase B is a functional component of an angiogenic switch during multistage pancreatic carcinogenesis by increasing the release of VEGF.
  • the present invention relates to methods of treating a human or animal subject with, and uses in a human or animal subject of, a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof.
  • the invention also relates to the use of the compound, tautomer, salt of the compound, salt of the tautomer, or the mixture thereof in the preparation of a medicament for use in the methods described herein.
  • the invention provides a method of modulating an inflammatory response or reducing cellular adhesion in a subject.
  • Such methods include administering to the subject a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof.
  • the inflammatory response is modulated in the subject and/or cellular adhesion is reduced in the subject after administration of the compound, the tautomer, the pharmaceutically acceptable salt of the compound, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof.
  • the compound, tautomer, salt of the compound, salt of the tautomer, or the mixture thereof are used to modulate an inflammatory response.
  • the compound, tautomer, salt of the compound, salt of the tautomer, or the mixture thereof are used to reduce cellular adhesion.
  • the compound, tautomer, salt of the compound, salt of the tautomer, or the mixture thereof are used to decrease ICAM, VCAM, or E- selectin levels.
  • the compound, tautomer, salt of the compound, salt of the tautomer, or the mixture thereof used to reduce the levels of circulating cell adhesion molecules.
  • the compound, tautomer, salt of the compound, salt of the tautomer, or the mixture thereof are used to decrease circulating ICAM, VCAM, or E-selectin levels.
  • the invention provides a method of monitoring the progression ofa disease or treatment in a human or animal subject.
  • the method includes measuring the amount of at least one cell adhesion molecule in the subject after administration of a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof to the subject.
  • the cell adhesion molecule is selected from inducible cell adhesion molecule (ICAM), vascular cell adhesion molecule (VCAM), or endothelial leukocyte adhesion molecule (E-Selectin).
  • Some such methods further include withdrawing a sample of blood from the subject and then measuring the amount of the at least one cell adhesion molecule in at least a portion of the sample.
  • Other embodiments include administering the compound, the tautomer, the pharmaceutically acceptable salt of the compound, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof to the subject.
  • the invention provides a method of identifying a subject in need of a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof.
  • the method includes measuring the amount of at least one cell adhesion molecule in the subject before, during, or after administration of the compound of Structure I, the tautomer of the compound, the pharmaceutically acceptable salt of the compound, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof to the subject.
  • the cell adhesion molecule is selected from inducible cell adhesion molecule, vascular cell adhesion molecule, or endothelial leukocyte adhesion molecule.
  • the method further includes administering the compound of Structure I, the tautomer of the compound, the pharmaceutically acceptable salt of the compound, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof to the subject after measuring the amount of the cell adhesion molecule in the subject.
  • Structure I has the following formula:
  • R 1 , R 2 , R 3 , and R 4 may be the same or different and are independently selected from the group consisting of H, Cl, Br, F, I, -CN, -NO 2 , -OH, -OR 15 groups, -NR 16 R 17 groups, substituted and unsubstituted amidinyl groups, substituted and unsubstituted guanidinyl groups, substituted and unsubstituted primary, secondary, and tertiary alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted alkenyl groups, substituted and unsubstituted alkynyl groups, substituted and unsubstituted heterocyclyl groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylamino
  • Figure 1 is a graph showing the effects of various amount of 4-amino-
  • Figures 2A and 2B are graphs showing the dose-dependent reduction of soluble ICAM ( Figure 2 A; greater than 70% inhibition with 100 or 150 mg/kg) and soluble VCAM ( Figure 2B; 44-47% inhibition with 100 or 150 mg/kg) in the serum of mice with 4T1 breast tumors when dosed with varying amounts of 4-amino-5-fluoro- 3-[6-(4-mefhylpiperazin-l-yl)-lH-benzimidazol-2-yl]quinolin-2(lH)-one.
  • Figure 3 is a graph showing the dose-dependent inhibition of mouse- specific soluble E-selectin in the serum of 4T1 tumor bearing mice treated with 4- amino-5 -fluoro-3 - [6-(4-methylpiperazin- 1 -yl)- 1 H-benzimidazol-2-yl] quinolin-2( 1 H)- one.
  • Figures 4A, 4B, and 4C are graphs of the Zymography and VEGF
  • Figure 5 is a scanned image showing the decrease in the expression of
  • Figure 6 is a scanned image showing the decrease in the expression of ⁇ 5 integrin, not ⁇ v integrin when HUVECs in culture were treated with 4-amino-5- fluoro-3 - [6-(4-methylpiperazin- 1 -yl)- 1 H-benzimidazol-2-y 1] quinolin-2( 1 H) .
  • cell adhesion refers to cell adhesion.
  • the amount of cellular adhesion in a subject can typically be correlated with the amounts of cell adhesion molecules, such as, but not limited to VCAM, ICAM, and E- Selectin in a subject.
  • VCAM is an abbreviation that stands for vascular cell adhesion molecule.
  • ICAM inducible cell adhesion molecule
  • E-Selectin is also known as endothelial leukocyte adhesion molecule.
  • 4T1 is a murine breast cell line.
  • BALB/C is a mice strain used in tumor xenograph experiments.
  • bFGF is an abbreviation that stands for basic fibroblast growth factor.
  • FGFR1 also referred to as bFGFR, is an abbreviation that stands for a tyrosine kinase that interacts with the fibroblast growth factor FGF.
  • FGF is an abbreviation for the fibroblast growth factor that interacts with FGFRl.
  • FGFR3 is an abbreviation that stands for the tyrosine kinase fibroblast growth factor receptor 3 that is often expressed in multiple myeloma-type cancers.
  • Flk-1 is an abbreviation that stands for fetal liver tyrosine kinase 1, also known as kinase-insert domain tyrosine kinase or KDR (human), also known as vascular endothelial growth factor receptor-2 or VEGFR2 (KDR (human), Flk-1 (mouse)).
  • PDGF is an abbreviation that stands for platelet derived growth factor. PDGF interacts with tyrosine kinases PDGFR ⁇ and PDGFR ⁇ .
  • RTK receptor tyrosine kinase
  • VEGF is an abbreviation that stands for vascular endothelial growth factor.
  • VEGF-RTK is an abbreviation that stands for vascular endothelial growth factor receptor tyrosine kinase.
  • ELISA is an abbreviation that stands for Enzyme-Linked
  • MMP-2 is an abbreviation that stands for matrix metalloprotease-2
  • MMP-2 is also referred to as gelatinase A.
  • MMP-9 is an abbreviation that stands for matrix metalloprotease-9
  • MMP-9 is also referred to as gelatinase B.
  • Ki67 is a marker for cellular proliferation.
  • caspase-3 is a apoptosis marker. Activation of caspase-3 requires proteolytic processing of inactive caspase-3 into “cleaved caspase-3" which is 17 KD and 19 KD in size.
  • PARP is an abbreviation that stands for poly ADP -ribose polymerase and is an apoptosis marker. It is a 116 KD protein and is cleaved into a 89KD protein.
  • CD31 is a marker for endothelial cells. Immuno staining with anti-
  • CD31 antibody in tumor section by immunohistochemistry will indicate the number of microvessels (or microvessel density) in tumors.
  • reference to a certain element such as hydrogen or H is meant to include all isotopes of that element.
  • an R group is defined to include hydrogen or H, it also includes deuterium and tritium.
  • unsubstituted alkyl refers to alkyl groups that do not contain heteroatoms.
  • the phrase includes straight chain alkyl groups such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl and the like.
  • the phrase also includes branched chain isomers of straight chain alkyl groups, including but not limited to, the following which are provided by way of example: -CH(CH 3 ) 2 , -CH(CH 3 )(CH 2 CH 3 ), -CH(CH 2 CH 3 ) 2 , -C(CH 3 ) 3 , -C(CH 2 CH 3 ) 3 , -CH 2 CH(CH 3 ) 2 , -CH 2 CH(CH 3 )(CH 2 CH 3 ), -CH 2 CH(CH 2 CH 3 ) 2 , -CH 2 C(CH 3 ) 3 , -CH 2 C(CH 2 CH 3 ) 3 , -CH(CH 3 )CH(CH 3 )(CH 2 CH 3 ), -CH 2 CH 2 CH(CH 3 ) 2 , -CH 2 CH 2 CH(CH 3 )(CH 2 CH 3 ), -CH 2 CH 2 CH(CH 3 ) 2 , -CH 2 CH(CH 3 )(CH 2
  • the phrase also includes cyclic alkyl groups such as cycloalkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl and such rings substituted with straight and branched chain alkyl groups as defined above.
  • cyclic alkyl groups such as cycloalkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl and such rings substituted with straight and branched chain alkyl groups as defined above.
  • the phrase also includes polycychc alkyl groups such as, but not limited to, adamantyl norbornyl, and bicyclo[2.2.2]octyl and such rings substituted with straight and branched chain alkyl groups as defined above.
  • unsubstituted alkyl groups includes primary
  • Unsubstituted alkyl groups may be bonded to one or more carbon atom(s), oxygen atom(s), nitrogen atom(s), and/or sulfur atom(s) in the parent compound.
  • Preferred unsubstituted alkyl groups include straight and branched chain alkyl groups and cyclic alkyl groups having 1 to 20 carbon atoms. More preferred such unsubstituted alkyl groups have from 1 to 10 carbon atoms while even more preferred such groups have from 1 to 5 carbon atoms.
  • Most preferred unsubstituted alkyl groups include straight and branched chain alkyl groups having from 1 to 3 carbon atoms and include methyl, ethyl, propyl, and -CH(CH 3 ) 2 .
  • substituted alkyl refers to an unsubstituted alkyl group as defined above in which one or more bonds to a carbon(s) or hydrogen(s) are replaced by a bond to non-hydrogen and non-carbon atoms such as, but not limited to, a halogen atom in halides such as F, Cl, Br, and I; an oxygen atom in groups such as hydroxyl groups, alkoxy groups, aryloxy groups, and ester groups; a sulfur atom in groups such as thiol groups, alkyl and aryl sulfide groups, sulfone groups, sulfonyl groups, and sulfoxide groups; a nitrogen atom in groups such as amines, amides, alkylamines, dialkylamines, arylamines, alkylarylamines, diarylamines, N-oxides, imides, and enamines; a silicon atom in groups such as in trialky
  • Substituted alkyl groups also include groups in which one or more bonds to a carbon(s) or hydrogen(s) atom is replaced by a bond to a heteroatom such as oxygen in carbonyl, carboxyl, and ester groups; nitrogen in groups such as imines, oximes, hydrazones, and nitriles.
  • Preferred substituted alkyl groups include, among others, alkyl groups in which one or more bonds to a carbon or hydrogen atom is/are replaced by one or more bonds to fluorine atoms.
  • One example of a substituted alkyl group is the trifluoromefhyl group and other alkyl groups that contain the trifluoromethyl group.
  • alkyl groups include those in which one or more bonds to a carbon or hydrogen atom is replaced by a bond to an oxygen atom such that the substituted alkyl group contains a hydroxyl, alkoxy, aryloxy group, or heterocyclyloxy group.
  • Still other alkyl groups include alkyl groups that have an amine, alkylamine, dialkylamine, arylamine, (alkyl)(aryl)amine, diarylamine, heterocyclylamine, (alkyl)(heterocyclyl)amine, (aryl)(heterocyclyl)amine, or diheterocyclylamine group.
  • unsubstituted aryl refers to aryl groups that do not contain heteroatoms.
  • the phrase includes, but is not limited to, groups such as phenyl, biphenyl, anthracenyl, naphthenyl by way of example.
  • the phrase "unsubstituted aryl” includes groups containing condensed rings such as naphthalene, it does not include aryl groups that have other groups such as alkyl or halo groups bonded to one of the ring members, as aryl groups such as tolyl are considered herein to be substituted aryl groups as described below.
  • a preferred unsubstituted aryl group is phenyl.
  • Unsubstituted aryl groups may be bonded to one or more carbon atom(s), oxygen atom(s), nitrogen atom(s), and/or sulfur atom(s) in the parent compound, however.
  • substituted aryl group has the same meaning with respect to unsubstituted aryl groups that substituted alkyl groups had with respect to unsubstituted alkyl groups.
  • a substituted aryl group also includes aryl groups in which one of the aromatic carbons is bonded to one of the non-carbon or non-hydrogen atoms described above and also includes aryl groups in which one or more aromatic carbons of the aryl group is bonded to a substituted and/or unsubstituted alkyl, alkenyl, or alkynyl group as defined herein.
  • unsubstituted alkenyl refers to straight and branched chain and cyclic groups such as those described with respect to unsubstituted alkyl groups as defined above, except that at least one double bond exists between two carbon atoms.
  • substituted alkenyl has the same meaning with respect to unsubstituted alkenyl groups that substituted alkyl groups had with respect to unsubstituted alkyl groups.
  • a substituted alkenyl group includes alkenyl groups in which a non-carbon or non-hydrogen atom is bonded to a carbon double bonded to another carbon and those in which one of the non-carbon or non-hydrogen atoms is bonded to a carbon not involved in a double bond to another carbon.
  • substituted alkynyl has the same meaning with respect to unsubstituted alkynyl groups that substituted alkyl groups had with respect to unsubstituted alkyl groups.
  • a substituted alkynyl group includes alkynyl groups in which a non-carbon or non-hydrogen atom is bonded to a carbon triple bonded to another carbon and those in which a non-carbon or non-hydrogen atom is bonded to a carbon not involved in a triple bond to another carbon.
  • unsubstituted aralkyl refers to unsubstituted alkyl groups as defined above in which a hydrogen or carbon bond of the unsubstituted alkyl group is replaced with a bond to an aryl group as defined above.
  • methyl (- CH 3 ) is an unsubstituted alkyl group.
  • a hydrogen atom of the methyl group is replaced by a bond to a phenyl group, such as if the carbon of the methyl were bonded to a carbon of benzene, then the compound is an unsubstituted aralkyl group (i.e., a benzyl group).
  • the phrase includes, but is not limited to, groups such as benzyl, diphenylmethyl, and 1-phenylethyl (-CH(C 6 H 5 )(CH3)) among others.
  • substituted aralkyl has the same meaning with respect to unsubstituted aralkyl groups that substituted aryl groups had with respect to unsubstituted aryl groups.
  • a substituted aralkyl group also includes groups in which a carbon or hydrogen bond of the alkyl part of the group is replaced by a bond to a non-carbon or a non-hydrogen atom. Examples of substituted aralkyl groups include, but are not limited to, and -CH 2 (2-methylphenyl) among others.
  • unsubstituted heterocyclyl refers to both aromatic and nonaromatic ring compounds including monocyclic, bicyclic, and polycychc ring compounds such as, but not limited to, quinuclidyl, containing 3 or more ring members of which one or more is a heteroatom such as, but not limited to, N, O, and S.
  • unsubstituted heterocyclyl includes condensed heterocyclic rings such as benzimidazolyl, it does not include heterocyclyl groups that have other groups such as alkyl or halo groups bonded to one of the ring members as compounds such as 2-methylbenzimidazolyl are substituted heterocyclyl groups.
  • heterocyclyl groups include, but are not limited to: unsaturated 3 to 8 membered rings containing 1 to 4 nitrogen atoms such as, but not limited to pyrrolyl, pyrrolinyl, imidazolyl, pyrazolyl, pyridinyl, dihydropyridinyl, pyrimidyl, pyrazinyl, pyridazinyl, triazolyl (e.g. 4H-l,2,4-triazolyl, lH-l,2,3-triazolyl, 2H-l,2,3-triazolyl etc.), tetrazolyl, (e.g.
  • saturated 3 to 8 membered rings containing 1 to 4 nitrogen atoms such as, but not limited to, pyrrolidinyl, imidazolidinyl, piperidinyl, piperazinyl; condensed unsaturated heterocyclic groups containing 1 to 4 nitrogen atoms such as, but not limited to, indolyl, isoindolyl, indolinyl, indolizinyl, benzimidazolyl, quinolyl, isoquinolyl, indazolyl, benzotriazolyl; unsaturated 3 to 8 membered rings containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms such as, but not limited to, oxazolyl, isoxazolyl, oxadiazolyl (e.g.
  • unsaturated 3 to 8 membered rings containing 1 to 3 sulfur atoms and 1 to 3 nitrogen atoms such as, but not limited to, thiazolyl, isothiazolyl, thiadiazolyl (e.g.
  • Heterocyclyl group also include those described above in which one or more S atoms in the ring is double-bonded to one or two oxygen atoms (sulfoxides and sulfones).
  • heterocyclyl groups include tetrahydrothiophene oxide and tetrahydrothiophene 1,1 -dioxide.
  • Preferred heterocyclyl groups contain 5 or 6 ring members.
  • More preferred heterocyclyl groups include morpholine, piperazine, piperidine, pyrrolidine, imidazole, pyrazole, 1,2,3-triazole, 1 ,2,4-triazole, tetrazole, thiophene, thiomorpholine, thiomorpholine in which the S atom of the thiomorpholine is bonded to one or more O atoms, pyrrole, homopiperazine, oxazolidin-2-one, pyrrolidin-2-one, oxazole, quinuclidine, thiazole, isoxazole, furan, and tetrahydrofuran.
  • substituted heterocyclyl refers to an unsubstituted heterocyclyl group as defined above in which one or more of the ring members is bonded to a non-hydrogen atom such as described above with respect to substituted alkyl groups and substituted aryl groups.
  • examples include, but are not limited to, 2- methylbenzimidazolyl, 5-methylbenzimidazolyl, 5-chlorobenzthiazolyl, N-alkyl piperazinyl groups such as 1-methyl piperazinyl, piperazine-N-oxide, N-alkyl piperazine N-oxides, 2-phenoxy-thiophene, and 2-chloropyridinyl among others.
  • substituted heterocyclyl groups also include heterocyclyl groups in which the bond to the non-hydrogen atom is a bond to a carbon atom that is part of a substituted and unsubstituted aryl, substituted and unsubstituted aralkyl, or unsubstituted heterocyclyl group.
  • Examples include but are not limited to 1- benzylpiperidinyl, 3-phenythiomorpholinyl, 3-(pyrrolidin-l-yl)-pyrrolidinyl, and 4- (piperidin-l-yl)-piperidinyl.
  • Groups such as N-alkyl substituted piperazine groups such as N-methyl piperazine, substituted morpholine groups, and piperazine N-oxide groups such as piperazine N-oxide and N-alkyl piperazine N-oxides are examples of some substituted heterocyclyl groups.
  • Groups such as substituted piperazine groups such as N-alkyl substituted piperazine groups such as N-methyl piperazine and the like, substituted morpholine groups, piperazine N-oxide groups, and N-alkyl piperazine N-oxide groups are examples of some substituted heterocyclyl groups that are especially suited as R 6 or R 7 groups.
  • unsubstituted heterocyclylalkyl refers to unsubstituted alkyl groups as defined above in which a hydrogen or carbon bond of the unsubstituted alkyl group is replaced with a bond to a heterocyclyl group as defined above.
  • methyl (-CH3) is an unsubstituted alkyl group. If a hydrogen atom of the methyl group is replaced by a bond to a heterocyclyl group, such as if the carbon of the methyl were bonded to carbon 2 of pyridine (one of the carbons bonded to the N of the pyridine) or carbons 3 or 4 of the pyridine, then the compound is an unsubstituted heterocyclylalkyl group.
  • substituted heterocyclylalkyl has the same meaning with respect to unsubstituted heterocyclylalkyl groups that substituted aralkyl groups had with respect to unsubstituted aralkyl groups.
  • a substituted heterocyclylalkyl group also includes groups in which a non-hydrogen atom is bonded to a heteroatom in the heterocyclyl group of the heterocyclylalkyl group such as, but not limited to, a nitrogen atom in the piperidine ring of a piperidinylalkyl group.
  • a substituted heterocyclylalkyl group also includes groups in which a carbon bond or a hydrogen bond of the alkyl part of the group is replaced by a bond to a substituted and unsubstituted aryl or substituted and unsubstituted aralkyl group. Examples include but are not limited to phenyl-(piperidin-l-yl)-methyl and phenyl-(morpholin-4-yl)- methyl.
  • unsubstituted alkylaminoalkyl refers to an unsubstituted alkyl group as defined above in which a carbon or hydrogen bond is replaced by a bond to a nitrogen atom that is bonded to a hydrogen atom and an unsubstituted alkyl group as defined above.
  • methyl (-CH 3 ) is an unsubstituted alkyl group. If a hydrogen atom of the methyl group is replaced by a bond to a nitrogen atom that is bonded to a hydrogen atom and an ethyl group, then the resulting compound is -CH 2 -N(H)(CH 2 CH3) which is an unsubstituted alkylaminoalkyl group.
  • substituted alkylaminoalkyl refers to an unsubstituted alkylaminoalkyl group as defined above except where one or more bonds to a carbon or hydrogen atom in one or both of the alkyl groups is replaced by a bond to a non- carbon or non-hydrogen atom as described above with respect to substituted alkyl groups except that the bond to the nitrogen atom in all alkylaminoalkyl groups does not by itself qualify all alkylaminoalkyl groups as being substituted.
  • substituted alkylaminoalkyl groups does include groups in which the hydrogen bonded to the nitrogen atom of the group is replaced with a non-carbon and non- hydrogen atom.
  • unsubstituted dialkylaminoalkyl refers to an unsubstituted alkyl group as defined above in which a carbon bond or hydrogen bond is replaced by a bond to a nitrogen atom which is bonded to two other similar or different unsubstituted alkyl groups as defined above.
  • substituted dialkylaminoalkyl refers to an unsubstituted dialkylaminoalkyl group as defined above in which one or more bonds to a carbon or hydrogen atom in one or more of the alkyl groups is replaced by a bond to a non- carbon and non-hydrogen atom as described with respect to substituted alkyl groups.
  • the bond to the nitrogen atom in all dialkylaminoalkyl groups does not by itself qualify all dialkylaminoalkyl groups as being substituted.
  • unsubstituted alkoxy refers to a hydroxyl group (-OH) in which the bond to the hydrogen atom is replaced by a bond to a carbon atom of an otherwise unsubstituted alkyl group as defined above.
  • substituted alkoxy refers to a hydroxyl group (-OH) in which the bond to the hydrogen atom is replaced by a bond to a carbon atom of an otherwise substituted alkyl group as defined above.
  • substituted heterocyclyloxy refers to a hydroxyl group (-
  • unsubstituted heterocyclyloxyalkyl refers to an unsubstituted alkyl group as defined above in which a carbon bond or hydrogen bond is replaced by a bond to an oxygen atom which is bonded to an unsubstituted heterocyclyl group as defined above.
  • substituted heterocyclyloxyalkyl refers to an unsubstituted heterocyclyloxyalkyl group as defined above in which a bond to a carbon or hydrogen group of the alkyl group of the heterocyclyloxyalkyl group is bonded to a non-carbon and non-hydrogen atom as described above with respect to substituted alkyl groups or in which the heterocyclyl group of the heterocyclyloxyalkyl group is a substituted heterocyclyl group as defined above.
  • unsubstituted heterocyclylalkoxy refers to an unsubstituted alkyl group as defined above in which a carbon bond or hydrogen bond is replaced by a bond to an oxygen atom which is bonded to the parent compound, and in which another carbon or hydrogen bond of the unsubstituted alkyl group is bonded to an unsubstituted heterocyclyl group as defined above.
  • substituted heterocyclylalkoxy refers to an unsubstituted heterocyclylalkoxy group as defined above in which a bond to a carbon or hydrogen group of the alkyl group of the heterocyclylalkoxy group is bonded to a non-carbon and non-hydrogen atom as described above with respect to substituted alkyl groups or in which the heterocyclyl group of the heterocyclylalkoxy group is a substituted heterocyclyl group as defined above.
  • a substituted heterocyclylalkoxy group also includes groups in which a carbon bond or a hydrogen bond to the alkyl moiety of the group may be substituted with one or more additional substituted and unsubstituted heterocycles. Examples include but are not limited to pyrid-2- ylmorpholin-4-ylmethyl and 2-pyrid-3-yl-2-morpholin-4-ylethyl.
  • unsubstituted arylaminoalkyl refers to an unsubstituted alkyl group as defined above in which a carbon bond or hydrogen bond is replaced by a bond to a nitrogen atom which is bonded to at least one unsubstituted aryl group as defined above.
  • substituted arylaminoalkyl refers to an unsubstituted arylaminoalkyl group as defined above except where either the alkyl group of the arylaminoalkyl group is a substituted alkyl group as defined above or the aryl group of the arylaminoalkyl group is a substituted aryl group except that the bonds to the nitrogen atom in all arylaminoalkyl groups does not by itself qualify all arylaminoalkyl groups as being substituted.
  • substituted arylaminoalkyl groups does include groups in which the hydrogen bonded to the nitrogen atom of the group is replaced with a non-carbon and non-hydrogen atom.
  • unsubstituted heterocyclylaminoalkyl refers to an unsubstituted alkyl group as defined above in which a carbon or hydrogen bond is replaced by a bond to a nitrogen atom which is bonded to at least one unsubstituted heterocyclyl group as defined above.
  • substituted heterocyclylaminoalkyl refers to unsubstituted heterocyclylaminoalkyl groups as defined above in which the heterocyclyl group is a substituted heterocyclyl group as defined above and/or the alkyl group is a substituted alkyl group as defined above.
  • the bonds to the nitrogen atom in all heterocyclylaminoalkyl groups does not by itself qualify all heterocyclylaminoalkyl groups as being substituted.
  • substituted heterocyclylaminoalkyl groups do include groups in which the hydrogen bonded to the nitrogen atom of the group is replaced with a non-carbon and non-hydrogen atom.
  • unsubstituted alkylaminoalkoxy refers to an unsubstituted alkyl group as defined above in which a carbon or hydrogen bond is replaced by a bond to an oxygen atom which is bonded to the parent compound and in which another carbon or hydrogen bond of the unsubstituted alkyl group is bonded to a nitrogen atom which is bonded to a hydrogen atom and an unsubstituted alkyl group as defined above.
  • substituted alkylaminoalkoxy refers to unsubstituted alkylaminoalkoxy groups as defined above in which a bond to a carbon or hydrogen atom of the alkyl group bonded to the oxygen atom which is bonded to the parent compound is replaced by one or more bonds to a non-carbon and non-hydrogen atoms as discussed above with respect to substituted alkyl groups and/or if the hydrogen bonded to the amino group is bonded to a non-carbon and non-hydrogen atom and/or if the alkyl group bonded to the nitrogen of the amine is bonded to a non-carbon and non-hydrogen atom as described above with respect to substituted alkyl groups.
  • unsubstituted dialkylaminoalkoxy refers to an unsubstituted alkyl group as defined above in which a carbon or hydrogen bond is replaced by a bond to an oxygen atom which is bonded to the parent compound and in which another carbon or hydrogen bond of the unsubstituted alkyl group is bonded to a nitrogen atom which is bonded to two other similar or different unsubstituted alkyl groups as defined above.
  • substituted dialkylaminoalkoxy refers to an unsubstituted dialkylaminoalkoxy group as defined above in which a bond to a carbon or hydrogen atom of the alkyl group bonded to the oxygen atom which is bonded to the parent compound is replaced by one or more bonds to a non-carbon and non-hydrogen atoms as discussed above with respect to substituted alkyl groups and/or if one or more of the alkyl groups bonded to the nitrogen of the amine is bonded to a non-carbon and non-hydrogen atom as described above with respect to substituted alkyl groups.
  • the presence of the amine and alkoxy functionality in all dialkylaminoalkoxy groups does not by itself qualify all such groups as substituted dialkylaminoalkoxy groups.
  • protected with respect to hydroxyl groups, amine groups, and sulfhydryl groups refers to forms of these functionalities which are protected from undesirable reaction with a protecting group known to those skilled in the art such as those set forth in Protective Groups in Organic Synthesis, Greene, T.W.; Wuts, P. G. M., John Wiley & Sons, New York, NY, (3rd Edition, 1999) which can be added or removed using the procedures set forth therein.
  • Examples of protected hydroxyl groups include, but are not limited to, silyl ethers such as those obtained by reaction of a hydroxyl group with a reagent such as, but not limited to, t-butyldimethyl- chlorosilane, trimefhylchlorosilane, triisopropylchlorosilane, triethylchlorosilane; substituted methyl and ethyl ethers such as, but not limited to methoxymethyl ether, methythiomethyl ether, benzyloxymethyl ether, /-butoxymefhyl ether, 2- methoxyethoxymethyl ether, tetrahydropyranyl ethers, 1-ethoxyethyl ether, allyl ether, benzyl ether; esters such as, but not limited to, benzoylformate, formate, acetate, trichloroacetate, and trifluoroacetate.
  • protected amine groups include, but are not limited to, amides such as, formamide, acetamide, trifluoroacetamide, and benzamide; imides, such as phfhalimide, and dithiosuccinimide; and others.
  • protected sulfhydryl groups include, but are not limited to, thioefhers such as S-benzyl thioether, and S-4-picolyl thioether; substituted S-methyl derivatives such as hemithio, dithio and aminothio acetals; and others.
  • a "pharmaceutically acceptable salt” includes a salt with an inorganic base, organic base, inorganic acid, organic acid, or basic or acidic amino acid.
  • the invention includes, for example, alkali metals such as sodium or potassium; alkaline earth metals such as calcium and magnesium or aluminum; and ammonia.
  • the invention includes, for example, trimethylamine, triethylamine, pyridine, picoline, ethanolamine, diethanolamine, and triethanolamine.
  • the instant invention includes, for example, hydrochloric acid, hydroboric acid, nitric acid, sulfuric acid, and phosphoric acid.
  • the instant invention includes, for example, formic acid, acetic acid, trifluoroacetic acid, fumaric acid, oxalic acid, tartaric acid, lactic acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, and p-toluenesulfonic acid.
  • salts of basic amino acids the instant invention includes, for example, arginine, lysine and ornithine.
  • Acidic amino acids include, for example, aspartic acid and glutamic acid.
  • the invention provides a method of modulating an inflammatory response and/or reducing cellular adhesion in a subject.
  • Such methods include administering to the subject a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof.
  • the inflammatory response is modulated in the subject and/or cellular adhesion is reduced in the subject after administration of the compound, the tautomer, the pharmaceutically acceptable salt of the compound, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof.
  • the invention provides a method of treating a disorder related to inflammation in a human or animal subject.
  • the method includes admimstering to the human or animal subject an effective amount of a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof.
  • Inflammation and inflammatory responses may occur with various biological conditions. Examples of such biological conditions may include cancer, autoimmune diseases, asthma, allergies, eczema, microbial infections, traumatic injuries such as burns or cuts, lupus, arthritis, cardiovascular disease such as, but not limited to, strokes and ischemic injuries, respiratory bacterial and viral infections, and other conditions associated with inflammatory responses.
  • the invention provides a method of treating a disorder related to cellular adhesion in a human or animal subject.
  • the method includes administering to the human or animal subject an effective amount ofa compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof.
  • the invention provides a method of decreasing cellular adhesion molecules such as ICAM, VCAM, E-selectin, MMP-2, or MMP-9 levels in a human or animal subject.
  • the method includes administering to the human or animal subject a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof.
  • the amount of the cellular adhesion molecule is typically reduced in the subject after administration.
  • the invention provides a method of decreasing circulating ICAM, VCAM, E-selectin, MMP-2, or MMP-9 levels in a human or animal subject.
  • the method includes administering to the human or animal subject a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof.
  • the invention provides a method of decreasing circulating cell adhesion molecules in a human or animal subject.
  • the method includes administering to the human or animal subject a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof.
  • the invention provides a method of monitoring the progression ofa disease or treatment in a human or animal subject.
  • the method includes administering to the human or animal subject a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof and measuring the amounts of a molecule such as ICAM, VCAM, E-selectin, MMP-2, or MMP-9 levels in the subject.
  • the invention provides a method of monitoring the progression of a disease or treatment in a human or animal subject.
  • the method includes measuring the amount of at least one cell adhesion molecule in the subject after administration of a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof to the subject.
  • the cell adhesion molecule is selected from inducible cell adhesion molecule (ICAM), vascular cell adhesion molecule (VCAM), or endothelial leukocyte adhesion molecule (E-Selectin).
  • Some such methods further include withdrawing a sample of blood from the subject and then measuring the amount of the at least one cell adhesion molecule in at least a portion of the sample.
  • the invention provides a method of identifying a subject in need of a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof.
  • the method includes measuring the amount of at least one cell adhesion molecule in the subject before, during, or after administration of the compound of Structure I, the tautomer of the compound, the pharmaceutically acceptable salt of the compound, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof to the subject.
  • the cell adhesion molecule is selected from inducible cell adhesion molecule, vascular cell adhesion molecule, or endothelial leukocyte adhesion molecule.
  • the cell adhesion molecule is selected from inducible cell adhesion molecule (ICAM), vascular cell adhesion molecule (VCAM), or endothelial leukocyte adhesion molecule (E-Selectin).
  • ICM inducible cell adhesion molecule
  • VCAM vascular cell adhesion molecule
  • E-Selectin endothelial leukocyte adhesion molecule
  • the subject is a cancer patient.
  • Structure I has the following formula:
  • R 1 , R 2 , R 3 , and R 4 may be the same or different and are independently selected from the group consisting of H, Cl, Br, F, I, -CN, -NO 2 , -OH, -OR 15 groups, -NR 16 R 17 groups, substituted and unsubstituted amidinyl groups, substituted and unsubstituted guanidinyl groups, substituted and unsubstituted primary, secondary, and tertiary alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted alkenyl groups, substituted and unsubstituted alkynyl groups, substituted and unsubstituted heterocyclyl groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylamino
  • R 5 , R 6 , R 7 , or R 8 is selected from the group consisting of substituted and unsubstituted amidinyl groups, substituted and unsubstituted guanidinyl groups, substituted and unsubstituted saturated heterocyclyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted heterocyclylalkyl groups, substituted and unsubstituted heterocyclylaminoalkyl groups, substituted and unsubstituted
  • the invention relates to a pharmaceutically acceptable salt of 4-amino-5-fluoro-3-[6-(4-methylpiperazin-l -yl)-lH-benzimidazol- 2-yl]quinolin-2(lH)-one (Compound 1) or a tautomer thereof.
  • the salt is selected from the group consisting of tartrate, malate, lactate, bis-acetate, citrate, mesylate, bismesylate and bishydrochloride.
  • the compound of structure I is a lactate salt of 4-amino-5-fluoro-3-[6-(4-methylpiperazin- 1 -yl)- 1 H-benzimidazol-2- yl]quinolin-2(lH)-one or a tautomer thereof.
  • the pharmaceutically acceptable salt of the compound of Structure I, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof is administered to the subject, and the salt is a lactate salt.
  • R 12 and R 13 are H, and in other embodiments, both R 12 and R 13 are H.
  • R 1 is selected from the group consisting of F,
  • substituted and unsubstituted alkoxy groups substituted and unsubstituted heterocyclylalkoxy groups, substituted and unsubstituted heterocyclyl groups, substituted and unsubstituted alkylaminoalkoxy groups, substituted and unsubstituted arylaminoalkoxy groups, substituted and unsubstituted dialkylaminoalkoxy groups, substituted and unsubstituted diarylaminoalkoxy groups, and substituted and unsubstituted (alkyl)(aryl)aminoalkoxy groups.
  • R 1 is F and R 2 , R 3 , R 3 , R 4 , R 5 , and R 8 are all H, and one of R 6 or R 7 is H.
  • At least one of R 5 , R 6 , R 7 , and R 8 is a substituted or unsubstituted heterocyclyl group.
  • At least one of R 5 , R 6 , R 7 , and R 8 is a substituted or unsubstituted heterocyclyl group comprising at least one O or N atom.
  • At least one of R 5 , R 6 , R 7 , and R 8 is a substituted or unsubstituted heterocyclyl group and the heterocyclyl group is selected from the group consisting of morpholine, piperazine, piperidine, pyrrolidine, thiomorpholine, homopiperazine, tetrahydrothiophene, tetrahydrofuran, and tetrahydropyran.
  • At least one of R 6 or R 7 is a substituted or unsubstituted heterocyclyl group.
  • At least one of R 6 or R 7 is a substituted or unsubstituted heterocyclyl group comprising at least one O or N atom.
  • one of R 6 or R 7 is a substituted or unsubstituted heterocyclyl group and the heterocyclyl group is selected from the group consisting of morpholine, piperazine, piperidine, pyrrolidine, thiomorpholine, homopiperazine, tetrahydrothiophene, tetrahydrofuran, and tetrahydropyran.
  • one of R 6 or R 7 is selected from the group consisting of substituted and unsubstituted morpholine groups, and substituted and unsubstituted piperazine groups.
  • one of R 6 or R 7 is a piperazine N-oxide or is an N-alkyl substituted piperazine.
  • R 6 or R 7 is selected from the group consisting of-NR 20 R 21 groups wherein R 20 is selected from the group consisting of substituted and unsubstituted heterocyclyl groups; and -NR 20 R 21 groups wherein R 21 is selected from the group consisting of substituted and unsubstituted heterocyclyl groups, groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted heterocyclylaminoalkyl groups, substituted and unsubstituted hydroxyalkyl groups
  • R 1 is selected from the group consisting of
  • the compounds and their corresponding salts and tautomers are provided in the following two tables below.
  • the synthesis of these compounds is described in U.S. Patent No. 6,605,617, published U.S. Patent Application No. 2004/0092535, published U.S. Patent Application No. 2004/0220196 as are various kinase assay procedures.
  • Each of these references is, therefore, hereby incorporated by reference in its entirety and for all purposes as if set forth in its entirety.
  • the compound of Structure I is a compound of Structure II, where Structure II has the following formula:
  • A is a group having one of the following Structures:
  • R a is selected from H or straight or branched chain alkyl groups having from 1 to 6 carbon atoms.
  • R a is a methyl group
  • the compound of Structure II is a compound of Structure II A
  • the pharmaceutically acceptable salt of the compound of Structure IIA, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof is administered to the subject, and the salt is a lactate salt.
  • R a is a H
  • the compound of Structure II is a compound of Structure IIB
  • R a is a methyl group
  • the compound of Structure II is a compound of Structure IIC
  • the compounds of any of the embodiments may be used to prepare medicaments or pharmaceutical formulations for use in any of the methods of the invention.
  • compositions for use with the invention may include any of the compounds, tautomers, or salts of any of the embodiments described above in combination with a pharmaceutically acceptable carrier such as those described herein.
  • compositions which may be prepared by mixing one or more compounds of the instant invention, or pharmaceutically acceptable salts tautomers thereof, or mixtures thereof with pharmaceutically acceptable carriers, excipients, binders, diluents or the like to treat or ameliorate disorders related to metastacized tumors.
  • the compositions of the inventions may be used to create formulations for use in any of the methods of the invention.
  • Such compositions can be in the form of, for example, granules, powders, tablets, capsules, syrup, suppositories, injections, emulsions, elixirs, suspensions or solutions.
  • compositions can be formulated for various routes of administration, for example, by oral administration, by nasal administration, by rectal administration, subcutaneous injection, intravenous injection, intramuscular injections, or intraperitoneal injection.
  • routes of administration for example, by oral administration, by nasal administration, by rectal administration, subcutaneous injection, intravenous injection, intramuscular injections, or intraperitoneal injection.
  • dosage forms are given by way of example and should not be construed as limiting the instant invention.
  • powders, suspensions, granules, tablets, pills, capsules, gelcaps, and caplets are acceptable as solid dosage forms. These can be prepared, for example, by mixing one or more compounds of the instant invention, pharmaceutically acceptable salts, tautomers, or mixtures thereof, with at least one additive such as a starch or other additive.
  • Suitable additives are sucrose, lactose, cellulose sugar, mannitol, maltitol, dextran, starch, agar, alginates, chitins, chitosans, pectins, tragacanth gum, gum arabic, gelatins, collagens, casein, albumin, synthetic or semi-synthetic polymers or glycerides.
  • oral dosage forms can contain other ingredients to aid in administration, such as an inactive diluent, or lubricants such as magnesium stearate, or preservatives such as paraben or sorbic acid, or anti-oxidants such as ascorbic acid, tocopherol or cysteine, a disintegrating agent, binders, thickeners, buffers, sweeteners, flavoring agents or perfuming agents. Tablets and pills may be further treated with suitable coating materials known in the art.
  • suitable coating materials known in the art.
  • Liquid dosage forms for oral administration may be in the form of pharmaceutically acceptable emulsions, syrups, elixirs, suspensions, and solutions, which may contain an inactive diluent, such as water.
  • Pharmaceutical formulations and medicaments may be prepared as liquid suspensions or solutions using a sterile liquid, such as, but not limited to, an oil, water, an alcohol, and combinations of these.
  • Pharmaceutically suitable surfactants, suspending agents, emulsifying agents may be added for oral or parenteral administration.
  • suspensions may include oils.
  • oils include, but are not limited to, peanut oil, sesame oil, cottonseed oil, corn oil and olive oil.
  • Suspension preparation may also contain esters of fatty acids such as ethyl oleate, isopropyl myristate, fatty acid glycerides and acetylated fatty acid glycerides.
  • Suspension formulations may include alcohols, such as, but not limited to, ethanol, isopropyl alcohol, hexadecyl alcohol, glycerol and propylene glycol.
  • Ethers such as but not limited to, poly(ethyleneglycol), petroleum hydrocarbons such as mineral oil and petrolatum; and water may also be used in suspension formulations.
  • the pharmaceutical formulations and medicaments may be a spray or aerosol containing an appropriate solvent(s) and optionally other compounds such as, but not limited to, stabilizers, antimicrobial agents, antioxidants, pH modifiers, surfactants, bioavailability modifiers and combinations of these.
  • a propellant for an aerosol formulation may include compressed air, nitrogen, carbon dioxide, or a hydrocarbon based low boiling solvent.
  • Injectable dosage forms generally include aqueous suspensions or oil suspensions which may be prepared using a suitable dispersant or wetting agent and a suspending agent. Injectable forms may be in solution phase or in the form of a suspension, which is prepared with a solvent or diluent. Acceptable solvents or vehicles include sterilized water, Ringer's solution, or an isotonic aqueous saline solution. Alternatively, sterile oils may be employed as solvents or suspending agents.
  • the oil or fatty acid is non- volatile, including natural or synthetic oils, fatty acids, mono-, di- or tri-glycerides.
  • the pharmaceutical formulation and/or medicament may be a powder suitable for reconstitution with an appropriate solution as described above.
  • these include, but are not limited to, freeze dried, rotary dried or spray dried powders, amorphous powders, granules, precipitates, or particulates.
  • the formulations may optionally contain stabilizers, pH modifiers, surfactants, bioavailability modifiers and combinations of these.
  • the pharmaceutical formulations and medicaments may be in the form of a suppository, an ointment, an enema, a tablet or a cream for release of compound in the intestines, sigmoid flexure and/or rectum.
  • Rectal suppositories are prepared by mixing one or more compounds of the instant invention, or pharmaceutically acceptable salts or tautomers of the compound, with acceptable vehicles, for example, cocoa butter or polyethylene glycol, which is present in a solid phase at normal storing temperatures, and present in a liquid phase at those temperatures suitable to release a drug inside the body, such as in the rectum.
  • Oils may also be employed in the preparation of formulations of the soft gelatin type and suppositories.
  • Water, saline, aqueous dextrose and related sugar solutions, and glycerols may be employed in the preparation of suspension formulations which may also contain suspending agents such as pectins, carbomers, methyl cellulose, hydroxypropyl cellulose or carboxymethyl cellulose, as well as buffers and preservatives.
  • excipients and carriers are generally known to those skilled in the art and are thus included in the instant invention. Such excipients and carriers are described, for example, in "Remingtons Pharmaceutical Sciences” Mack Pub. Co., New Jersey (1991), which is incorporated herein by reference in its entirety for all purposes as if fully set forth herein.
  • the formulations of the invention may be designed to be short-acting, fast-releasing, long-acting, and sustained-releasing as described below.
  • the pharmaceutical formulations may also be formulated for controlled release or for slow release.
  • compositions may also comprise, for example, micelles or liposomes, or some other encapsulated form, or may be administered in an extended release form to provide a prolonged storage and/or delivery effect. Therefore, the pharmaceutical formulations and medicaments may be compressed into pellets or cylinders and implanted intramuscularly or subcutaneously as depot injections or as implants such as stents. Such implants may employ known inert materials such as silicones and biodegradable polymers.
  • Specific dosages may be adjusted depending on conditions of disease, the age, body weight, general health conditions, sex, and diet of the subject, dose intervals, administration routes, excretion rate, and combinations of drugs. Any of the above dosage forms containing effective amounts are well within the bounds of routine experimentation and therefore, well within the scope of the instant invention.
  • a therapeutically effective dose may vary depending upon the route of administration and dosage form.
  • the preferred compound or compounds of the instant invention is a formulation that exhibits a high therapeutic index.
  • the therapeutic index is the dose ratio between toxic and therapeutic effects which can be expressed as the ratio between LD50 and ED50.
  • the LD50 is the dose lethal to 50% of the population and the ED 50 is the dose therapeutically effective in 50% of the population.
  • the LD 5 o and ED 5 o are determined by standard pharmaceutical procedures in animal cell cultures or experimental animals.
  • compositions and medicaments according to the invention include the compound of Structure I or the tautomers, salts, or mixtures thereof in combination with a pharmaceutically acceptable carrier.
  • the compounds of the invention may be used to prepare medicaments and pharmaceutical formulations.
  • Such medicaments and pharmaceutical formulations may be used in any of the methods of treatment described herein.
  • kinase inhibitors for use as anticancer agents in conjunction with the methods or compositions of the present invention include inhibitors of Epidermal Growth Factor Receptor (EGFR) kinases such as small molecule quinazolines, for example gefitinib (US 5457105, US 5616582, and US 5770599), ZD-6474 (WO 01/32651), erlotinib (Tarceva®, US 5,747,498 and WO 96/30347), and lapatinib (US 6,727,256 and WO 02/02552).
  • EGFR Epidermal Growth Factor Receptor
  • Kinase inhibitors for use as anticancer agents in conjunction with the methods of compositions of the present invention also include inibitors of Vascular Endothelial Growth Factor Receptor (VEGFR) kinase inhibitor such as, but not limited to, SU- 11248 (WO 01/60814), SU 5416 (US 5,883,113 and WO 99/61422), SU 6668 (US 5,883,113 and WO 99/61422), CHIR-258 (US 6,605,617 and US 6,774,237), vatalanib or PTK-787 (US 6,258,812), VEGF-Trap (WO 02/57423), B43-Genistein (WO-09606116), fenre inide (retinoic acid p-hydroxyphenylamine) (US 4,323,581), IM-862 (WO 02/62826), bevacizumab or Avastin® (WO 94/10202), KRN-951, 3-[5- (methylsulfonylpiperadine
  • the compounds of the invention may be used to treat a variety of subjects.
  • Suitable subjects include animals such as mammals and humans.
  • Suitable mammals include, but are not limited to, primates such as, but not limited to lemurs, apes, and monkeys; rodents such as rats, mice, and guinea pigs; rabbits and hares; cows; horses; pigs; goats; sheep; marsupials; and carnivores such as felines, canines, and ursines.
  • the subject or patient is a human.
  • the subject or patient is a rodent such as a mouse or a rat.
  • the subject or patient is an animal other than a human and in some such embodiments, the subject or patient is a mammal other than a human.
  • HPLC solvents were from Burdick and Jackson (Muskegan, Michigan), or Fisher Scientific (Pittsburg, Pennsylvania). In some instances, purity was assessed by thin layer chromatography (TLC) using glass or plastic backed silica gel plates, such as, for example, Baker-Flex Silica Gel 1B2-F flexible sheets. TLC results were readily detected visually under ultraviolet light, or by employing well known iodine vapor and other various staining techniques.
  • Mass spectrometric analysis was performed on one of two LCMS instruments: a Waters System (Alliance HT HPLC and a Micromass ZQ mass spectrometer; Column: Eclipse XDB-C18, 2.1 x 50 mm; Solvent system: 5-95% acetonitrile in water with 0.05% TFA; Flow rate 0.8 mL/minute; Molecular weight range 150-850; Cone Voltage 20 V; Column temperature 40°C) or a Hewlett Packard System (Series 1100 HPLC; Column: Eclipse XDB-C18, 2.1 x 50 mm; Solvent system: 1-95%) acetonitrile in water with 0.05%> TFA; Flow rate 0.4 mL/minute; Molecular weight range 150-850; Cone Voltage 50 V; Column temperature 30°C). All masses are reported as those of the protonated parent ions. [0139] GCMS analysis was performed on a Hewlet Packard instrument
  • Preparative separations were carried out using either a Flash 40 chromatography system and KP-Sil, 60A (Biotage, Charlottesville, Virginia), or by HPLC using a C-18 reversed phase column.
  • Typical solvents employed for the Flash 40 Biotage system were dichloromethane, methanol, ethyl acetate, hexane and triethyl amine.
  • Typical solvents employed for the reverse phase HPLC were varying concentrations of acetonitrile and water with 0.1% trifluoroacetic acid.
  • the resulting mixture was filtered, and the resulting solid was washed with TBME (500 mL, 2X) and then was dried under vacuum for one hour using a rubber dam.
  • the resulting solid was transferred to a drying tray and dried in a vacuum oven at 50°C to a constant weight to yield 670 g (97.8%) of the title compound as a yellow powder.
  • the resulting mixture was then filtered, and the flask and filter cake were washed with water (3 x 2.56 L).
  • the golden yellow solid product was dried to a constant weight of 416 g (98.6%) yield) under vacuum at about 50°C in a vacuum oven.
  • the flask was heated in a heating mantle to an internal temperature of 97°C (+/- 5°C) and maintained at that temperature until the reaction was complete (typically about 40 hours) as determined by HPLC. After the reaction was complete, heating was discontinued and the reaction was cooled to an internal temperature of about 80°C with stirring, and water (3.15 L) was added to the mixture via an addition funnel over the period of 1 hour while the internal temperature was maintained at 82°C (+/- 3°C). After water addition was complete, heating was discontinued and the reaction mixture was allowed to cool over a period of no less than 4 hours to an internal temperature of 20-25°C. The reaction mixture was then stirred for an additional hour at an internal temperature of 20-30°C.
  • the resulting mixture was then filtered, and the flask and filter cake were washed with water (l x l L), 50% ethanol (1 x IL), and 95% ethanol (1 x IL).
  • the golden yellow solid product was placed in a drying pan and dried to a constant weight of 546 g (99% yield) under vacuum at about 50°C in a vacuum oven.
  • a 5000 mL, 4-neck flask was fitted with a stirrer, thermometer, condenser, and gas inlet/outlet.
  • the equipped flask was charged with 265.7 g (1.12 mol. 1.0 eq) of 5-(4-methyl-piperazin-l-yl)-2-nitroaniline and 2125 mL of 200 proof EtOH.
  • the resulting solution was purged with N 2 for 15 minutes.
  • 20.0 g of 5% Pd/C (50% H O w/w) was added.
  • the reaction was vigorously stirred at 40-50°C (internal temperature) while H 2 was bubbled through the mixture.
  • the reaction was monitored hourly for the disappearance of 5-(4-methyl-piperazin-l-yl)-2-nitroaniline by HPLC.
  • the typical reaction time was 6 hours.
  • a 5000 mL, 4-neck jacketed flask was fitted with a mechanical stirrer, condenser, temperature probe, gas inlet, and oil bubbler.
  • the equipped flask was charged with 300 g (1.27 mol) of 5-(4-methyl-piperazin-l-yl)-2-nitroaniline and 2400 mL of 200 proof EtOH (the reaction may be and has been conducted with 95% ethanol and it is not necessary to use 200 proof ethanol for this reaction).
  • the resulting solution was stirred and purged with N 2 for 15 minutes.
  • 22.7 g of 5% Pd/C (50% H 2 O w/w) was added to the reaction flask.
  • the reaction vessel was purged with N 2 for 15 minutes.
  • reaction vessel was purged with H 2 by maintaining a slow, but constant flow of H 2 through the flask.
  • the reaction was stirred at 45-55°C (internal temperature) while H 2 was bubbled through the mixture until the 5-(4-methyl-piperazin-l-yl)-2-nitroaniline was completely consumed as determined by HPLC.
  • the typical reaction time was 6 hours.
  • the bright yellow solid was placed in a drying tray and dried in a vacuum oven at 50°C overnight providing 155.3 g (47.9%) of the desired 4-amino-5-fTuoro-3-[6-(4- mefhyl-piperazin- 1 -yl)- 1 H-benzimidazol-2-yl] - 1 H-quinolin-2-one .
  • a 5000 mL 4-neck jacketed flask was equipped with a distillation apparatus, a temperature probe, a N 2 gas inlet, an addition funnel, and a mechanical stirrer.
  • [6-(4-Methyl-piperazin-l-yl)-lH-benzimidazol-2-yl]-acetic acid ethyl ester (173.0 g, 570 mmol) was charged into the reactor, and the reactor was purged with N 2 for 15 minutes. Dry THF (2600 mL) was then charged into the flask with stirring. After all the solid had dissolved, solvent was removed by distillation (vacuum or atmospheric (the higher temperature helps to remove the water) using heat as necessary.
  • the reaction was stirred for 3.5 to 4.5 hours (in some examples it was stirred for 30 to 60 minutes and the reaction may be complete within that time) while maintaining the internal temperature at from 38- 42°C. A sample of the reaction was then removed and analyzed by HPLC. If the reaction was not complete, additional KHMDS solution was added to the flask over a period of 5 minutes and the reaction was stirred at 38-42°C for 45-60 minutes (the amount of KHMDS solution added was determined by the following: If the IPC ratio is ⁇ 3.50, then 125 mL was added; if 10.0 > IPC ratio > 3.50, then 56 mL was added; if 20.0 > IPC ratio > 10, then 30 mL was added.
  • the IPC ratio is equal to the area corresponding to 4-amino-5-fluoro-3-[6-(4-methyl-piperazin-l-yl)-lH-benzimidazol- 2-yl]-lH-quinolin-2-one) divided by the area corresponding to the uncyclized intermediate).
  • the reflux condenser was then replaced with a distillation apparatus and solvent was removed by distillation (vacuum or atmospheric) using heat as required. After 1500 mL of solvent had been removed, distillation was discontinued and the reaction was purged with N 2 . Water (1660 mL) was then added to the reaction flask while maintaining the internal temperature at 20-30°C. The reaction mixture was then stirred at 20-30°C for 30 minutes before cooling it to an internal temperature of 5- 10°C and then stirring for 1 hour. The resulting suspension was filtered, and the flask and filter cake were washed with water (3 x 650 mL).
  • the internal temperature of the mixture was raised until a temperature of 63°C (+/- 3°C) was achieved.
  • the reaction was then monitored for completion using HPLC to check for consumption of the starting materials (typically in 2-3 hours, both starting materials were consumed (less than 0.5% by area % HPLC)). If the reaction was not complete after 2 hours, another 0.05 equivalents of potassium t- butoxide was added at a time, and the process was completed until HPLC showed that the reaction was complete. After the reaction was complete, 650 mL of water was added to the stirred reaction mixture. The reaction was then warmed to an internal temperature of 50°C and the THF was distilled away (about 3 L by volume) under reduced pressure from the reaction mixture. Water (2.6 L) was then added dropwise to the reaction mixture using an addition funnel. The mixture was then cooled to room temperature and stirred for at least 1 hour.
  • a 3000 mL 4-neck flask equipped with a condenser, temperature probe, N 2 gas inlet, and mechanical stirrer was placed in a heating mantle.
  • the flask was then charged with 4-amino-5-fluoro-3-[6-(4-methyl-piperazin-l-yl)-lH- benzimidazol-2-yl]-lH-quinolin-2-one (101.0 g, 0.26 mol), and the yellow solid was suspended in 95% ethanol (1000 mL) and stirred. In some cases an 8:1 solvent ratio is used.
  • the suspension was then heated to a gentle reflux (temperature of about
  • a 3000 mL 4-necked jacketed flask was fitted with a condenser, a temperature probe, a N 2 gas inlet, and a mechanical stirrer.
  • the reaction vessel was purged with N 2 for at least 15 minutes and then charged with 4-amino-5-fluoro-3-[6- (4-methyl-piperazin-l-yl)-lH-benzimidazol-2-yl]-lH-quinolin-2-one (484 g, 1.23 mol).
  • a solution of D,L-Lactic acid 243.3 g, 1.72 mol of monomer-see the following paragraph
  • water (339 mL)
  • ethanol (1211 mL) was prepared and then charged to the reaction flask.
  • the reaction flask was then cooled to an internal temperature ranging from about 64-70°C within 15-25 minutes and this temperature was maintained for a period of about 30 minutes.
  • the reactor was inspected for crystals. If no crystals were present, then crystals of the lactic acid salt of 4-amino-5-fluoro-3- [6-(4-methyl-piperazin- 1 -yl)- 1 H-benzimidazol -2-yl] - 1 H- quinolin-2-one (484 mg, 0.1 mole %) were added to the flask, and the reaction was stirred at 64-70°C for 30 minutes before again inspecting the flask for crystals.
  • the collected solid was dried to a constant weight at 50°C under vacuum in a vacuum oven yielding 510.7 g (85.7%) of the crystalline yellow lactic acid salt of 4-amino-5-fluoro-3-[6-(4-methyl-piperazin- l-yl)-lH-benzimidazol-2-yl]-lH-quinolin-2-one.
  • a rubber dam or inert conditions were typically used during the filtration process. While the dry solid did not appear to be very hygroscopic, the wet filter cake tends to pick up water and become sticky. Precautions were taken to avoid prolonged exposure of the wet filter cake to the atmosphere.
  • lactic acid generally contains about 8-12% w/w water, and contains dimers and trimers in addition to the monomeric lactic acid.
  • the mole ratio of lactic acid dimer to monomer is generally about 1.0:4.7.
  • Commercial grade lactic acid may be used in the process described in the preceding paragraph as the monolactate salt preferentially precipitates from the reaction mixture.
  • Compound 2 the N-oxide metabolite of Compound 1, was synthesized as shown in the scheme below.
  • Compound 1 was heated in a mixture of ethanol, dimethylacetamide and hydrogen peroxide.
  • Compound 2 was isolated by filtration and washed with ethanol. If necessary, the product could be further purified by column chromatography.
  • Compound 3 the N-desmethyl metabolite of Compound 1, was synthesized as shown in the scheme below. 5-Chloro-2-nitroaniline was treated with piperazine to yield 4 which was subsequently protected with a butyloxycarbonyl (Boc) group to yield 5. Reduction of the nitro group followed by condensation with 3-ethoxy-3-iminopropionic acid ethyl ester gave 6. Condensation of 6 with 6- fluoroanthranilonitrile using potassium hexamethyldisilazide as the base yielded 7. Crude 7 was treated with aqueous HCl to yield the desired metabolite as a yellow/brown solid after purification.
  • 5-Chloro-2-nitroaniline was treated with piperazine to yield 4 which was subsequently protected with a butyloxycarbonyl (Boc) group to yield 5. Reduction of the nitro group followed by condensation with 3-ethoxy-3-iminopropionic acid ethyl ester gave 6. Condensation
  • 4T1 breast tumor cells were grown as subcutaneous tumors in BALB/C mice, and treatment (10, 30, 60, 100, and 150 mg/kg) with 4-amino-5-fluoro-3-[6-(4- methylpiperazin-l-yl)-lH-benzimidazol-2-yl]quinolin-2(lH)-one (Compound 1) were initiated when tumors were approximately 150 mm 3 . Mice were dosed orally, daily for 18 days.
  • FIG. 1 is a graph showing the effects of 4-amino-5-fluoro-3-[6-(4- methylpiperazin-l-yl)-lH-benzimidazol-2-yl]quinolin-2(lH)-one in the 4T1 murine breast tumor model.
  • the growth of subcutaneous tumors was inhibited (40-80% compared to control), liver metastases were completely inhibited, and lung metastases were inhibited by 60-97% after 18 days of dosing.
  • Table 1 Table 1
  • the standard was used at a range of 4000 pg/mL to 31 pg/mL.
  • the serum samples were diluted 1/200 followed by 3-fold serial dilutions.
  • the samples and standards were added at 50 ⁇ L/well and incubated at 37°C for 1 hour.
  • the plates were washed three times and incubated at 37°C for 1 hour with the primary antibody (biotinylated goat anti-mouse VCAM-1, R&D Systems #BAF643) diluted 1/200 in wash buffer, 50 ⁇ L/well.
  • the plates were washed as described above and incubated at 37°C for 1 hour with stepavidin-HRP (R&D Systems #DY998) 1/200 in PBS/1% goat serum without Tween 20.
  • the standard was used at a dilution range of 1/10-1/1280.
  • the serum samples were diluted 1/15 followed by 3-fold serial dilutions.
  • the samples and standards were added at 50 ⁇ L/well and incubated at 37°C for 1 hour.
  • the plates were washed three times and incubated at 37°C for 1 hour with the primary antibody (goat anti-ICAM-1, Santa Cruz Biotechnology #sc-1511) diluted 1/250 in wash buffer, 50 ⁇ L/well.
  • the plates were washed as above and incubated at 37°C for 1 hour with 50 ⁇ L/well of the secondary antibody (swine anti-goat IgG HRPO labeled, Caltag #G50007) 1/2000 in wash buffer.
  • mice Female Nu/nu mice (6-8 weeks old, 18-22 grams) were obtained from
  • Tumor cells (2 x 10 6 KM12L4a) were implanted subcutaneous into the flank of mice and allowed to grow to the desired size before treatment was initiated.
  • Tumor bearing mice were administered with 100 mg/kg of 4-amino-5-fluoro-3-[6-(4-methylpiperazin-l-yl)-lH-benzimidazol-2- yl]quinolin-2(lH)-one for 7 days, and individual mice were euthanized. The tumors were resected and flash frozen in liquid nitrogen.
  • Resected tumors were lysed in RIPA buffer (1 % Nonidet P-40, 0.5% sodium deoxycholate, 0.1% Sodium dodecylsulphate in IX phosphate buffered saline, pH 7.2) containing protease inhibitors (Roche Molecular Biochemicals) and phosphatase inhibitors (Sigma). 50 ⁇ g of total proteins were analyzed by gelatin zymography on 12% SDS polyacrylamide with gelatin substrate.
  • VEGF-A protein levels in KM12L4a tumor lysates were quantified using a commercially available ELISA kit (R and D Systems, Minneapolis, MN) according to the manufacturer's procedures.
  • HUVECs were cultured in EGM (Endothelial Cell Growth Media) with or without 100 nM 4-amino-5-fluoro-3-[6-(4-methylpiperazin-l-yl)-lH- benzimidazol-2-yl]quinolin-2(lH)-one (Compound 1), and cell lysates were collected at 0, 16, and 24 hours post-treatment. Equal amounts of proteins were loaded in 4- 20% SDS-PAGE, and the gels were probed with antibodies against ICAM, VCAM, ⁇ 5 integrin,, and ⁇ v integrin. The equal loading and efficiency was evaluated by probing with anti ⁇ -actin antibody.
  • the organic compounds according to the invention may exhibit the phenomenon of tautomerism.
  • the chemical stnictures within this specification can only represent one of the possible tautomeric forms at a time, it should be understood that the invention encompasses any tautomeric form of the drawn structure.
  • the compound of Structure IIIB is shown below with one tautomer, Tautomer IIIBa:

Landscapes

  • Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Rheumatology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Pain & Pain Management (AREA)
  • Oncology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Plural Heterocyclic Compounds (AREA)

Abstract

Methods of using compounds having Structure (I) or the salts or tautomers of the compounds in the treatment of disorders relating to cell adhesion and metastatic processes are presented herein.Formula (I) N HDisorders may include cancer, autoimmune diseases, asthma, allergies, eczema, microbial infections, traumatic injuries such as burns or cuts, lupus, arthritis, cardiovascular disease such as, but not limited to, strokes and ischemic injuries, respiratory bacterial and viral infections, and other conditions associated with inflammatory responses.

Description

MODULATION OF INFLAMMATORY AND METASTATIC PROCESSES
FIELD OF THE INVENTION
[0001] The present invention provides methods for using compounds to modulate inflammatory responses and processes related to tumor metastasis. The invention further provides methods for monitoring the effects of the compounds of the invention by measuring the levels of ICAM, VCAM, or E-selectin molecules in a subject treated with the compounds.
BACKGROUND OF THE INVENTION
[0002] Amino quinolinone benzimidazolyl compounds such as 4-amino-5- fluoro-3 - [6-(4-methylpiperazin- 1 -yl)- 1 H-benzimidazol-2-yl] quinolin-2( 1 H)-one and their tautomers and salts are potent inhibitors of various class kinases such as VEGFR2 (KDR, Flk-1), FGFR1 and PDGFRβ with IC50s ranging from 10-27 nM. See U.S. Patent No. 6,605,617, U.S. Patent Application No. 10/644,055, and U.S. Patent Application No. 10/706,328, each of which is hereby incorporated by reference in its entirety and for all purposes as if fully set forth herein, for a list of various tyrosine and serine/threonine kinases for which 4-amino-5-fluoro-3-[6-(4- methylpiperazin-l-yl)-lH-benzimidazol-2-yl]quinolin-2(lH)-one has shown activity and for assay procedures. Such kinases are important for the initiation and maintenance of new blood vessel growth as well as tumor proliferation. Consequently these inhibitors have direct applications in the treatment of various disorders such as solid and hematological cancers. The identification of plasma biomarkers in subjects treated with these kinase inhibitors would therefore provide a convenient method for monitoring the subject's physiological response to the treatment.
[0003] Cell adhesion molecules play important roles in tumor cell invasion, metastasis, and interaction with immune cells. NCAM (vascular cell adhesion molecule) is a transmembrane glycoprotein and expressed in endothelial cells and various cancer types such as bladder, breast, gastrointestinal, ovarian, renal, and Hodgkin's and non-Hodgkin's lymphoma. VCAM is induced by VEGF and is predominantly expressed in activated endofhelial cells. ICAM (inducible cell adhesion molecule) is also expressed in endothelial cells and various cells including fibroblasts, hematopoietic cells, and tumor cells. The soluble form of ICAM present in the plasma is generated by proteolytic cleavage of membrane-associated molecules. E-Selectin (endofhelial leukocyte adhesion molecule) is a transmembrane glycoprotein expressed in endothelial cells and mediates adhesion of neutrophils, monocytes, and T cells to endothelial cells. E-selectin also mediates tumor progression and metastasis.
[0004] A high concentration of soluble ICAM, VCAM, and E-selectin is considered a marker of endothelial cell activation during tumor development, metastasis, and inflammatory responses. These cell adhesion molecules localized on endothelial cells can mediate adhesion of metastatic tumor cells and allow extravasation into the vessels. It is of interest that these molecules are inducible, being poorly expressed on normal endothelial cells but capable of being expressed highly after exposure to cytokines such as IL-1 or TNF-a. In addition, some of these molecules are preferentially expressed in different vascular beds, with VCAM being abundant in the lung and E-selectin in the liver.
[0005] Matrix metalloproteases (MMPs) are a class of proteases that degrade most components of the extracellular matrix (ECM). Under normal physiological conditions they play an important role in development, tissue remodeling and morphogenesis. However, elevated levels of certain metalloproteases have been implicated in pathological diseases such as cancer and inflammation. Degradation of the extracellular matrix in the basement membrane is essential for tissue invasion by tumor cells and metastasis at various sites, and this degradation is dependent on the activity of MMPs. The family of MMPs includes more than 20 members. Two of these proteases are MMP-2 (gelatinase A, 72 KD) and MMP-9 (gelatinase B, 92 KD). MMP-2 and MMP-9 are important regulators of cancer progression and metastasis and their levels are frequently elevated in various cancer patients.
[0006] A report by Bergers et al. (Matrix metalloproteinase-9 triggers the angiogenic switch during carcinogenesis; Berger, G. et al, Nature Cell Biology, 2:737-744; 2000) discloses that MMP-9/gelatinase B is a functional component of an angiogenic switch during multistage pancreatic carcinogenesis by increasing the release of VEGF.
[0007] Various quinolinone benzimidazole compounds useful in inhibiting angiogenesis and vascular endothelial growth factor receptor tyrosine kinases and in inhibiting other tyrosine and serine/threonine kinases including 4-amino-5-fluoro-3- [5-(4-methylpiperazin-l-yl)-lH-benzimidazol-2-yl]quinolin-2(lH)-one or a tautomer thereof and the synthesis thereof are disclosed in the following documents which are each hereby incorporated by reference in their entireties and for all purposes as if fully set forth herein: U.S. Patent No. 6,605,617; U.S. Patent No. 6,756,383; U.S. Patent Application No. 10/116,117 filed (published on February 6, 2003, as US 2003/0028018 Al); U.S. Patent Application No. 10/644,055 (published on May 13, 2004, U.S. Patent Application No. 2004/0092535); U.S. Patent Application No. 10/983,174; U.S. Patent Application No. 10/706,328 (published on November 4, 2004, as 2004/0220196); U.S. Patent Application No. 10/982,757; and U.S. Patent Application No. 10/982,543.
[0008] An important need exists for methods for modulating levels of cellular adhesion molecules and matrix metalloproteases. Such methods would therefore constitute important and needed therapies in the treatment of inflammatory and metastatic diseases mediated by cellular adhesion molecules and matrix metalloproteases.
SUMMARY OF THE INVENTION
[0009] The present invention relates to methods of treating a human or animal subject with, and uses in a human or animal subject of, a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof. The invention also relates to the use of the compound, tautomer, salt of the compound, salt of the tautomer, or the mixture thereof in the preparation of a medicament for use in the methods described herein.
[0010] In one aspect, the invention provides a method of modulating an inflammatory response or reducing cellular adhesion in a subject. Such methods include administering to the subject a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof. The inflammatory response is modulated in the subject and/or cellular adhesion is reduced in the subject after administration of the compound, the tautomer, the pharmaceutically acceptable salt of the compound, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof.
[0011] In one aspect, the compound, tautomer, salt of the compound, salt of the tautomer, or the mixture thereof are used to modulate an inflammatory response.
[0012] In another aspect, the compound, tautomer, salt of the compound, salt of the tautomer, or the mixture thereof are used to reduce cellular adhesion.
[0013] In another aspect, the compound, tautomer, salt of the compound, salt of the tautomer, or the mixture thereof are used to decrease ICAM, VCAM, or E- selectin levels.
[0014] In another aspect, the compound, tautomer, salt of the compound, salt of the tautomer, or the mixture thereof used to reduce the levels of circulating cell adhesion molecules. [0015] In another aspect, the compound, tautomer, salt of the compound, salt of the tautomer, or the mixture thereof are used to decrease circulating ICAM, VCAM, or E-selectin levels.
[0016] In another aspect, the invention provides a method of monitoring the progression ofa disease or treatment in a human or animal subject. The method includes measuring the amount of at least one cell adhesion molecule in the subject after administration of a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof to the subject. In some embodiments, the cell adhesion molecule is selected from inducible cell adhesion molecule (ICAM), vascular cell adhesion molecule (VCAM), or endothelial leukocyte adhesion molecule (E-Selectin). Some such methods further include withdrawing a sample of blood from the subject and then measuring the amount of the at least one cell adhesion molecule in at least a portion of the sample. Other embodiments include administering the compound, the tautomer, the pharmaceutically acceptable salt of the compound, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof to the subject.
[0017] In another aspect, the invention provides a method of identifying a subject in need of a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof. The method includes measuring the amount of at least one cell adhesion molecule in the subject before, during, or after administration of the compound of Structure I, the tautomer of the compound, the pharmaceutically acceptable salt of the compound, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof to the subject. In some embodiments, the cell adhesion molecule is selected from inducible cell adhesion molecule, vascular cell adhesion molecule, or endothelial leukocyte adhesion molecule. In some embodiments, the method further includes administering the compound of Structure I, the tautomer of the compound, the pharmaceutically acceptable salt of the compound, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof to the subject after measuring the amount of the cell adhesion molecule in the subject. [0018] Structure I has the following formula:
Figure imgf000008_0001
wherein, R1, R2, R3, and R4 may be the same or different and are independently selected from the group consisting of H, Cl, Br, F, I, -CN, -NO2, -OH, -OR15 groups, -NR16R17 groups, substituted and unsubstituted amidinyl groups, substituted and unsubstituted guanidinyl groups, substituted and unsubstituted primary, secondary, and tertiary alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted alkenyl groups, substituted and unsubstituted alkynyl groups, substituted and unsubstituted heterocyclyl groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted heterocyclylalkyl groups, and -C(=O)R18 groups; R5, R6, R7, and R8 may be the same or different and are independently selected from the group consisting of H, Cl, Br, F, I, -NO2, -OH, -OR19 groups, -NR20R21 groups, -SH, -SR22 groups, -S(=O)R23 groups, -S(=O)2R24 groups, -CN, substituted and unsubstituted amidinyl groups, substituted and unsubstituted guanidinyl groups, substituted and unsubstituted primary, secondary, and tertiary alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted alkenyl groups, substituted and unsubstituted alkynyl groups, substituted and unsubstituted heterocyclyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted heterocyclylalkyl groups, -C(=O)R25 groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted heterocyclylaminoalkyl groups, substituted and unsubstituted hydroxyalkyl groups, substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted aryloxyalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups; R12 is selected from the group consisting of H, substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, and substituted and unsubstituted heterocyclyl groups; R13 is selected from the group consisting of H, substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted heterocyclyl groups, -OH, alkoxy groups, aryloxy groups, -NH , substituted and unsubstituted heterocyclylalkyl groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted alkylamino groups, substituted and unsubstituted arylamino groups, substituted and unsubstituted dialkylamino groups, substituted and unsubstituted diarylamino groups, substituted and unsubstituted (alkyl)(aryl)amino groups, -C(=O)H, -C(=O)-alkyl groups, -C(=O)-aryl groups, -C(=O)O-alkyl groups, -C(=O)O-aryl groups, -C(=0)NH2, -C(=O)NH(alkyl) groups, -C(=O)NH(aryl) groups, -C(=O)N(alkyl)2 groups, -C(=O)N(aryl)2 groups, -C(=O)N(alkyl)(aryl) groups, -C(=O)-heterocyclyl groups, -C(=O)-O-heterocyclyl groups, -C(=O)NH(heterocyclyl) groups, -C(=O)-N(heterocyclyl)2 groups, -C(=O)-N(alkyl)(heterocyclyl) groups, -C(=O)- N(aryl)(heterocyclyl) groups, substituted and unsubstituted heterocyclylaminoalkyl groups, substituted and unsubstituted hydroxyalkyl groups, substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted aryloxyalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups; R15 and R19 may be the same or different and are independently selected from the group consisting of substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted heterocyclyl groups, substituted and unsubstituted heterocyclylalkyl groups, -C(=O)H, -C(=O)-alkyl groups, -C(=O)-aryl groups, -C(=O)NH2, -C(=O)NH(alkyl) groups, -C(=O)NH(aryl) groups, -C(=O)N(alkyl)2 groups, -C(=O)N(aryl)2 groups, -C(=O)N(alkyl)(aryl) groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted heterocyclylaminoalkyl, substituted and unsubstituted diheterocyclylammoalkyl, substituted and unsubstituted (heterocyclyl)(alkyl)aminoalkyl, substituted and unsubstituted (heterocyclyl)(aryl)aminoalkyl, substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted hydroxyalkyl groups, substituted and unsubstituted aryloxyalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups; R16 and R20 may be the same or different and are independently selected from the group consisting of H, substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, and substituted and unsubstituted heterocyclyl groups; R17 and R21 may be the same or different and are independently selected from the group consisting of H, substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted heterocyclyl groups, -C(=O)H, -C(=O)-alkyl groups, -C(=O)-aryl groups,-C(=O)NH2, -C(=O)NH(alkyl) groups, -C(=O)NH(aryl) groups, -C(=O)N(alkyl)2 groups, -C(=O)N(aryl)2 groups, -C(=O)N(alkyl)(aryl) groups, -C(=O)O-alkyl groups, -C(=O)O-aryl groups, substituted and unsubstituted heterocyclylalkyl groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, -C(=O)-heterocyclyl groups, -C(=O)-O-heterocyclyl groups, -C(=O)NH(heterocyclyl) groups, -C(=O)-N(heterocyclyl)2 groups, -C(=O)-N(alkyl)(heterocyclyl) groups, -C(=O)-N(aryl)(heterocyclyl) groups, substituted and unsubstituted heterocyclylaminoalkyl groups, substituted and unsubstituted hydroxyalkyl groups, substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted aryloxyalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups; R18, R23, R24, and R25 may be the same or different and are independently selected from the group consisting of H, -NH2, -NH(alkyl) groups, -NH(aryl) groups, -N(alkyl)2 groups, -N(aryl)2 groups, -N(alkyl)(aryl) groups, -NH(heterocyclyl) groups, -N(heterocyclyl)(alkyl) groups, -N(heterocyclyl)(aryl) groups, -N(heterocyclyl)2 groups, substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, -OH, substituted and unsubstituted alkoxy groups, substituted and unsubstituted aryloxy groups, substituted and unsubstituted heterocyclyl groups, -NHOH, -N(alkyl)OH groups, -N(aryl)OH groups, -N(alkyl)O-alkyl groups, -N(aryl)O-alkyl groups, -N(alkyl)O-aryl groups, and -N(aryl)O-aryl groups; and R22 is selected from the group consisting of substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, and substituted and unsubstituted heterocyclyl groups.
[0019] Further objects, features and advantages of the invention will be apparent from the following detailed description.
BRIEF DESCRIPTION OF THE DRAWINGS
[0020] Figure 1 is a graph showing the effects of various amount of 4-amino-
5-fluoro-3-[6-(4-methylpiperazin-l-yl)-lH-benzimidazol-2-yl]quinolin-2(lH)-one (Compound 1) on a 4T1 murine breast tumor model (vehicle (grey outlined circle); 10 mpk (square); 30 mpk (grey triangle); 60 mpk (X); 100 mpk (diamond); and 150 mpk (filed circle)). The growth of the subcutaneous tumor was inhibited (40-80% compared to control), liver metastases were completely inhibited, and lung metastases were inhibited by 60-97%) after 18 days of dosing.
[0021] Figures 2A and 2B are graphs showing the dose-dependent reduction of soluble ICAM (Figure 2 A; greater than 70% inhibition with 100 or 150 mg/kg) and soluble VCAM (Figure 2B; 44-47% inhibition with 100 or 150 mg/kg) in the serum of mice with 4T1 breast tumors when dosed with varying amounts of 4-amino-5-fluoro- 3-[6-(4-mefhylpiperazin-l-yl)-lH-benzimidazol-2-yl]quinolin-2(lH)-one.
[0022] Figure 3 is a graph showing the dose-dependent inhibition of mouse- specific soluble E-selectin in the serum of 4T1 tumor bearing mice treated with 4- amino-5 -fluoro-3 - [6-(4-methylpiperazin- 1 -yl)- 1 H-benzimidazol-2-yl] quinolin-2( 1 H)- one.
[0023] Figures 4A, 4B, and 4C are graphs of the Zymography and VEGF
ELISA (Figure 4B) data that show the decrease in MMP9 and VEGF in mice with implanted KM12L4a tumor cells when dosed for 7 days with 100 mg/kg 4-amino-5- fluoro-3-[6-(4-methylpiperazin-l-yl)-lH-benzimidazol-2-yl]quinolin-2(lH)-one.
[0024] Figure 5 is a scanned image showing the decrease in the expression of
ICAM, and VCAM when HUVECs in culture were treated with 4-amino-5-fluoro-3- [6-(4-methylpiperazin- 1 -yl)- 1 H-benzimidazol-2-y 1] quinolin-2( 1 H) .
[0025] Figure 6 is a scanned image showing the decrease in the expression of α5 integrin, not αv integrin when HUVECs in culture were treated with 4-amino-5- fluoro-3 - [6-(4-methylpiperazin- 1 -yl)- 1 H-benzimidazol-2-y 1] quinolin-2( 1 H) .
DETAILED DESCRIPTION OF THE INVENTION
[0026] The following abbreviations and definitions are used throughout this application:
[0027] The phrase "cellular adhesion" as used herein, refers to cell adhesion.
The amount of cellular adhesion in a subject can typically be correlated with the amounts of cell adhesion molecules, such as, but not limited to VCAM, ICAM, and E- Selectin in a subject.
[0028] "VCAM" is an abbreviation that stands for vascular cell adhesion molecule.
[0029] "ICAM" is an abbreviation that stands for inducible cell adhesion molecule. [0030] "E-Selectin" is also known as endothelial leukocyte adhesion molecule.
[0031] "4T1" is a murine breast cell line.
[0032] "BALB/C" is a mice strain used in tumor xenograph experiments.
[0033] "bFGF" is an abbreviation that stands for basic fibroblast growth factor.
[0034] "FGFR1", also referred to as bFGFR, is an abbreviation that stands for a tyrosine kinase that interacts with the fibroblast growth factor FGF.
[0035] "FGF" is an abbreviation for the fibroblast growth factor that interacts with FGFRl.
[0036] "FGFR3" is an abbreviation that stands for the tyrosine kinase fibroblast growth factor receptor 3 that is often expressed in multiple myeloma-type cancers.
[0037] "Flk-1" is an abbreviation that stands for fetal liver tyrosine kinase 1, also known as kinase-insert domain tyrosine kinase or KDR (human), also known as vascular endothelial growth factor receptor-2 or VEGFR2 (KDR (human), Flk-1 (mouse)).
[0038] "PDGF" is an abbreviation that stands for platelet derived growth factor. PDGF interacts with tyrosine kinases PDGFRα and PDGFRβ.
[0039] "RTK" is an abbreviation that stands for receptor tyrosine kinase.
[0040] "VEGF" is an abbreviation that stands for vascular endothelial growth factor.
[0041] "VEGF-RTK" is an abbreviation that stands for vascular endothelial growth factor receptor tyrosine kinase. [0042] "ELISA" is an abbreviation that stands for Enzyme-Linked
Immunosorbent Assay.
[0043] "MMP-2" is an abbreviation that stands for matrix metalloprotease-2
[includes the 72 KD (pro MMP-2) protein and the 62 KD (active MMP-2) protein]. MMP-2 is also referred to as gelatinase A.
[0044] "MMP-9" is an abbreviation that stands for matrix metalloprotease-9
[includes the 105 KD (pro MMP-9) protein and the 92KD (active MMP-9) protein]. MMP-9 is also referred to as gelatinase B.
[0045] "Ki67" is a marker for cellular proliferation.
[0046] "Caspase-3" is a apoptosis marker. Activation of caspase-3 requires proteolytic processing of inactive caspase-3 into "cleaved caspase-3" which is 17 KD and 19 KD in size.
[0047] "PARP" is an abbreviation that stands for poly ADP -ribose polymerase and is an apoptosis marker. It is a 116 KD protein and is cleaved into a 89KD protein.
[0048] "CD31" is a marker for endothelial cells. Immuno staining with anti-
CD31 antibody in tumor section by immunohistochemistry will indicate the number of microvessels (or microvessel density) in tumors.
[0049] Generally, reference to a certain element such as hydrogen or H is meant to include all isotopes of that element. For example, if an R group is defined to include hydrogen or H, it also includes deuterium and tritium.
[0050] The phrase "unsubstituted alkyl" refers to alkyl groups that do not contain heteroatoms. Thus the phrase includes straight chain alkyl groups such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl and the like. The phrase also includes branched chain isomers of straight chain alkyl groups, including but not limited to, the following which are provided by way of example: -CH(CH3)2, -CH(CH3)(CH2CH3), -CH(CH2CH3)2, -C(CH3)3, -C(CH2CH3)3, -CH2CH(CH3)2, -CH2CH(CH3)(CH2CH3), -CH2CH(CH2CH3)2, -CH2C(CH3)3, -CH2C(CH2CH3)3, -CH(CH3)CH(CH3)(CH2CH3), -CH2CH2CH(CH3)2, -CH2CH2CH(CH3)(CH2CH3), -CH2CH2CH(CH2CH3)2, -CH2CH2C(CH3)3, -CH2CH2C(CH2CH3)3, -CH(CH3)CH2CH(CH3)2, -CH(CH3)CH(CH3)CH(CH3)2, -CH(CH2CH3)CH(CH3)CH(CH3)(CH2CH3), and others. The phrase also includes cyclic alkyl groups such as cycloalkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl and such rings substituted with straight and branched chain alkyl groups as defined above. The phrase also includes polycychc alkyl groups such as, but not limited to, adamantyl norbornyl, and bicyclo[2.2.2]octyl and such rings substituted with straight and branched chain alkyl groups as defined above. Thus, the phrase unsubstituted alkyl groups includes primary alkyl groups, secondary alkyl groups, and tertiary alkyl groups. Unsubstituted alkyl groups may be bonded to one or more carbon atom(s), oxygen atom(s), nitrogen atom(s), and/or sulfur atom(s) in the parent compound. Preferred unsubstituted alkyl groups include straight and branched chain alkyl groups and cyclic alkyl groups having 1 to 20 carbon atoms. More preferred such unsubstituted alkyl groups have from 1 to 10 carbon atoms while even more preferred such groups have from 1 to 5 carbon atoms. Most preferred unsubstituted alkyl groups include straight and branched chain alkyl groups having from 1 to 3 carbon atoms and include methyl, ethyl, propyl, and -CH(CH3)2.
[0051] The phrase "substituted alkyl" refers to an unsubstituted alkyl group as defined above in which one or more bonds to a carbon(s) or hydrogen(s) are replaced by a bond to non-hydrogen and non-carbon atoms such as, but not limited to, a halogen atom in halides such as F, Cl, Br, and I; an oxygen atom in groups such as hydroxyl groups, alkoxy groups, aryloxy groups, and ester groups; a sulfur atom in groups such as thiol groups, alkyl and aryl sulfide groups, sulfone groups, sulfonyl groups, and sulfoxide groups; a nitrogen atom in groups such as amines, amides, alkylamines, dialkylamines, arylamines, alkylarylamines, diarylamines, N-oxides, imides, and enamines; a silicon atom in groups such as in trialkylsilyl groups, dialkylarylsilyl groups, alkyldiarylsilyl groups, and triarylsilyl groups; and other heteroatoms in various other groups. Substituted alkyl groups also include groups in which one or more bonds to a carbon(s) or hydrogen(s) atom is replaced by a bond to a heteroatom such as oxygen in carbonyl, carboxyl, and ester groups; nitrogen in groups such as imines, oximes, hydrazones, and nitriles. Preferred substituted alkyl groups include, among others, alkyl groups in which one or more bonds to a carbon or hydrogen atom is/are replaced by one or more bonds to fluorine atoms. One example of a substituted alkyl group is the trifluoromefhyl group and other alkyl groups that contain the trifluoromethyl group. Other alkyl groups include those in which one or more bonds to a carbon or hydrogen atom is replaced by a bond to an oxygen atom such that the substituted alkyl group contains a hydroxyl, alkoxy, aryloxy group, or heterocyclyloxy group. Still other alkyl groups include alkyl groups that have an amine, alkylamine, dialkylamine, arylamine, (alkyl)(aryl)amine, diarylamine, heterocyclylamine, (alkyl)(heterocyclyl)amine, (aryl)(heterocyclyl)amine, or diheterocyclylamine group.
[0052] The phrase "unsubstituted aryl" refers to aryl groups that do not contain heteroatoms. Thus the phrase includes, but is not limited to, groups such as phenyl, biphenyl, anthracenyl, naphthenyl by way of example. Although the phrase "unsubstituted aryl" includes groups containing condensed rings such as naphthalene, it does not include aryl groups that have other groups such as alkyl or halo groups bonded to one of the ring members, as aryl groups such as tolyl are considered herein to be substituted aryl groups as described below. A preferred unsubstituted aryl group is phenyl. Unsubstituted aryl groups may be bonded to one or more carbon atom(s), oxygen atom(s), nitrogen atom(s), and/or sulfur atom(s) in the parent compound, however.
[0053] The phrase "substituted aryl group" has the same meaning with respect to unsubstituted aryl groups that substituted alkyl groups had with respect to unsubstituted alkyl groups. However, a substituted aryl group also includes aryl groups in which one of the aromatic carbons is bonded to one of the non-carbon or non-hydrogen atoms described above and also includes aryl groups in which one or more aromatic carbons of the aryl group is bonded to a substituted and/or unsubstituted alkyl, alkenyl, or alkynyl group as defined herein. This includes bonding arrangements in which two carbon atoms of an aryl group are bonded to two atoms of an alkyl, alkenyl, or alkynyl group to define a fused ring system (e.g. dihydronaphthyl or tetrahydronaphthyl). Thus, the phrase "substituted aryl" includes, but is not limited to tolyl, and hydroxyphenyl among others.
[0054] The phrase "unsubstituted alkenyl" refers to straight and branched chain and cyclic groups such as those described with respect to unsubstituted alkyl groups as defined above, except that at least one double bond exists between two carbon atoms. Examples include, but are not limited to vinyl, -CH=C(H)(CH3), -CH=C(CH3)2, -C(CH3)=C(H)2, -C(CH3)=C(H)(CH3), -C(CH2CH3)=CH2, cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl among others.
[0055] The phrase "substituted alkenyl" has the same meaning with respect to unsubstituted alkenyl groups that substituted alkyl groups had with respect to unsubstituted alkyl groups. A substituted alkenyl group includes alkenyl groups in which a non-carbon or non-hydrogen atom is bonded to a carbon double bonded to another carbon and those in which one of the non-carbon or non-hydrogen atoms is bonded to a carbon not involved in a double bond to another carbon.
[0056] The phrase "unsubstituted alkynyl" refers to straight and branched chain groups such as those described with respect to unsubstituted alkyl groups as defined above, except that at least one triple bond exists between two carbon atoms. Examples include, but are not limited to -C≡C(H), -C≡C(CH3), -C=C(CH2CH3), -C(H2)C≡C(H), -C(H)2C=C(CH3), and -C(H)2C≡C(CH2CH3) among others.
[0057] The phrase "substituted alkynyl" has the same meaning with respect to unsubstituted alkynyl groups that substituted alkyl groups had with respect to unsubstituted alkyl groups. A substituted alkynyl group includes alkynyl groups in which a non-carbon or non-hydrogen atom is bonded to a carbon triple bonded to another carbon and those in which a non-carbon or non-hydrogen atom is bonded to a carbon not involved in a triple bond to another carbon.
[0058] The phrase "unsubstituted aralkyl" refers to unsubstituted alkyl groups as defined above in which a hydrogen or carbon bond of the unsubstituted alkyl group is replaced with a bond to an aryl group as defined above. For example, methyl (- CH3) is an unsubstituted alkyl group. If a hydrogen atom of the methyl group is replaced by a bond to a phenyl group, such as if the carbon of the methyl were bonded to a carbon of benzene, then the compound is an unsubstituted aralkyl group (i.e., a benzyl group). Thus the phrase includes, but is not limited to, groups such as benzyl, diphenylmethyl, and 1-phenylethyl (-CH(C6H5)(CH3)) among others.
[0059] The phrase "substituted aralkyl" has the same meaning with respect to unsubstituted aralkyl groups that substituted aryl groups had with respect to unsubstituted aryl groups. However, a substituted aralkyl group also includes groups in which a carbon or hydrogen bond of the alkyl part of the group is replaced by a bond to a non-carbon or a non-hydrogen atom. Examples of substituted aralkyl groups include, but are not limited to,
Figure imgf000018_0001
and -CH2(2-methylphenyl) among others.
[0060] The phrase "unsubstituted heterocyclyl" refers to both aromatic and nonaromatic ring compounds including monocyclic, bicyclic, and polycychc ring compounds such as, but not limited to, quinuclidyl, containing 3 or more ring members of which one or more is a heteroatom such as, but not limited to, N, O, and S. Although the phrase "unsubstituted heterocyclyl" includes condensed heterocyclic rings such as benzimidazolyl, it does not include heterocyclyl groups that have other groups such as alkyl or halo groups bonded to one of the ring members as compounds such as 2-methylbenzimidazolyl are substituted heterocyclyl groups. Examples of heterocyclyl groups include, but are not limited to: unsaturated 3 to 8 membered rings containing 1 to 4 nitrogen atoms such as, but not limited to pyrrolyl, pyrrolinyl, imidazolyl, pyrazolyl, pyridinyl, dihydropyridinyl, pyrimidyl, pyrazinyl, pyridazinyl, triazolyl (e.g. 4H-l,2,4-triazolyl, lH-l,2,3-triazolyl, 2H-l,2,3-triazolyl etc.), tetrazolyl, (e.g. lH-tetrazolyl, 2H tetrazolyl, etc.); saturated 3 to 8 membered rings containing 1 to 4 nitrogen atoms such as, but not limited to, pyrrolidinyl, imidazolidinyl, piperidinyl, piperazinyl; condensed unsaturated heterocyclic groups containing 1 to 4 nitrogen atoms such as, but not limited to, indolyl, isoindolyl, indolinyl, indolizinyl, benzimidazolyl, quinolyl, isoquinolyl, indazolyl, benzotriazolyl; unsaturated 3 to 8 membered rings containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms such as, but not limited to, oxazolyl, isoxazolyl, oxadiazolyl (e.g. 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,5-oxadiazolyl, etc.); saturated 3 to 8 membered rings containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms such as, but not limited to, morpholinyl; unsaturated condensed heterocyclic groups containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms, for example, benzoxazolyl, benzoxadiazolyl, benzoxazinyl (e.g. 2H-l,4-benzoxazinyl etc.); unsaturated 3 to 8 membered rings containing 1 to 3 sulfur atoms and 1 to 3 nitrogen atoms such as, but not limited to, thiazolyl, isothiazolyl, thiadiazolyl (e.g. 1,2,3-thiadiazolyl, 1,2,4- thiadiazolyl, 1,3, 4-thiadiazolyl, 1,2,5-thiadiazolyl, etc.); saturated 3 to 8 membered rings containing 1 to 2 sulfur atoms and 1 to 3 nitrogen atoms such as, but not limited to, thiazolodinyl; saturated and unsaturated 3 to 8 membered rings containing 1 to 2 sulfur atoms such as, but not limited to, thienyl, dihydrodithiinyl, dihydrodithionyl, tetrahydrothiophene, tetrahydrothiopyran; unsaturated condensed heterocyclic rings containing 1 to 2 sulfur atoms and 1 to 3 nitrogen atoms such as, but not limited to, benzothiazolyl, benzothiadiazolyl, benzothiazinyl (e.g. 2H-l,4-benzothiazinyl, etc.), dihydrobenzothiazinyl (e.g., 2H-3,4-dihydrobenzothiazinyl, etc.), unsaturated 3 to 8 membered rings containing oxygen atoms such as, but not limited to furyl; unsaturated condensed heterocyclic rings containing 1 to 2 oxygen atoms such as benzodioxolyl (e.g., 1,3-benzodioxoyl, etc.); unsaturated 3 to 8 membered rings containing an oxygen atom and 1 to 2 sulfur atoms such as, but not limited to, dihydrooxathiinyl; saturated 3 to 8 membered rings containing 1 to 2 oxygen atoms and 1 to 2 sulfur atoms such as 1,4-oxathiane; unsaturated condensed rings containing 1 to 2 sulfur atoms such as benzothienyl, benzodithiinyl; and unsaturated condensed heterocyclic rings containing an oxygen atom and 1 to 2 oxygen atoms such as benzoxathiinyl. Heterocyclyl group also include those described above in which one or more S atoms in the ring is double-bonded to one or two oxygen atoms (sulfoxides and sulfones). For example, heterocyclyl groups include tetrahydrothiophene oxide and tetrahydrothiophene 1,1 -dioxide. Preferred heterocyclyl groups contain 5 or 6 ring members. More preferred heterocyclyl groups include morpholine, piperazine, piperidine, pyrrolidine, imidazole, pyrazole, 1,2,3-triazole, 1 ,2,4-triazole, tetrazole, thiophene, thiomorpholine, thiomorpholine in which the S atom of the thiomorpholine is bonded to one or more O atoms, pyrrole, homopiperazine, oxazolidin-2-one, pyrrolidin-2-one, oxazole, quinuclidine, thiazole, isoxazole, furan, and tetrahydrofuran.
[0061] The phrase "substituted heterocyclyl" refers to an unsubstituted heterocyclyl group as defined above in which one or more of the ring members is bonded to a non-hydrogen atom such as described above with respect to substituted alkyl groups and substituted aryl groups. Examples, include, but are not limited to, 2- methylbenzimidazolyl, 5-methylbenzimidazolyl, 5-chlorobenzthiazolyl, N-alkyl piperazinyl groups such as 1-methyl piperazinyl, piperazine-N-oxide, N-alkyl piperazine N-oxides, 2-phenoxy-thiophene, and 2-chloropyridinyl among others. In addition, substituted heterocyclyl groups also include heterocyclyl groups in which the bond to the non-hydrogen atom is a bond to a carbon atom that is part ofa substituted and unsubstituted aryl, substituted and unsubstituted aralkyl, or unsubstituted heterocyclyl group. Examples include but are not limited to 1- benzylpiperidinyl, 3-phenythiomorpholinyl, 3-(pyrrolidin-l-yl)-pyrrolidinyl, and 4- (piperidin-l-yl)-piperidinyl. Groups such as N-alkyl substituted piperazine groups such as N-methyl piperazine, substituted morpholine groups, and piperazine N-oxide groups such as piperazine N-oxide and N-alkyl piperazine N-oxides are examples of some substituted heterocyclyl groups. Groups such as substituted piperazine groups such as N-alkyl substituted piperazine groups such as N-methyl piperazine and the like, substituted morpholine groups, piperazine N-oxide groups, and N-alkyl piperazine N-oxide groups are examples of some substituted heterocyclyl groups that are especially suited as R6 or R7 groups.
[0062] The phrase "unsubstituted heterocyclylalkyl" refers to unsubstituted alkyl groups as defined above in which a hydrogen or carbon bond of the unsubstituted alkyl group is replaced with a bond to a heterocyclyl group as defined above. For example, methyl (-CH3) is an unsubstituted alkyl group. If a hydrogen atom of the methyl group is replaced by a bond to a heterocyclyl group, such as if the carbon of the methyl were bonded to carbon 2 of pyridine (one of the carbons bonded to the N of the pyridine) or carbons 3 or 4 of the pyridine, then the compound is an unsubstituted heterocyclylalkyl group. [0063] The phrase "substituted heterocyclylalkyl" has the same meaning with respect to unsubstituted heterocyclylalkyl groups that substituted aralkyl groups had with respect to unsubstituted aralkyl groups. However, a substituted heterocyclylalkyl group also includes groups in which a non-hydrogen atom is bonded to a heteroatom in the heterocyclyl group of the heterocyclylalkyl group such as, but not limited to, a nitrogen atom in the piperidine ring of a piperidinylalkyl group. In addition, a substituted heterocyclylalkyl group also includes groups in which a carbon bond or a hydrogen bond of the alkyl part of the group is replaced by a bond to a substituted and unsubstituted aryl or substituted and unsubstituted aralkyl group. Examples include but are not limited to phenyl-(piperidin-l-yl)-methyl and phenyl-(morpholin-4-yl)- methyl.
[0064] The phrase "unsubstituted alkylaminoalkyl" refers to an unsubstituted alkyl group as defined above in which a carbon or hydrogen bond is replaced by a bond to a nitrogen atom that is bonded to a hydrogen atom and an unsubstituted alkyl group as defined above. For example, methyl (-CH3) is an unsubstituted alkyl group. If a hydrogen atom of the methyl group is replaced by a bond to a nitrogen atom that is bonded to a hydrogen atom and an ethyl group, then the resulting compound is -CH2-N(H)(CH2CH3) which is an unsubstituted alkylaminoalkyl group.
[0065] The phrase "substituted alkylaminoalkyl" refers to an unsubstituted alkylaminoalkyl group as defined above except where one or more bonds to a carbon or hydrogen atom in one or both of the alkyl groups is replaced by a bond to a non- carbon or non-hydrogen atom as described above with respect to substituted alkyl groups except that the bond to the nitrogen atom in all alkylaminoalkyl groups does not by itself qualify all alkylaminoalkyl groups as being substituted. However, substituted alkylaminoalkyl groups does include groups in which the hydrogen bonded to the nitrogen atom of the group is replaced with a non-carbon and non- hydrogen atom.
[0066] The phrase "unsubstituted dialkylaminoalkyl" refers to an unsubstituted alkyl group as defined above in which a carbon bond or hydrogen bond is replaced by a bond to a nitrogen atom which is bonded to two other similar or different unsubstituted alkyl groups as defined above.
[0067] The phrase "substituted dialkylaminoalkyl" refers to an unsubstituted dialkylaminoalkyl group as defined above in which one or more bonds to a carbon or hydrogen atom in one or more of the alkyl groups is replaced by a bond to a non- carbon and non-hydrogen atom as described with respect to substituted alkyl groups. The bond to the nitrogen atom in all dialkylaminoalkyl groups does not by itself qualify all dialkylaminoalkyl groups as being substituted.
[0068] The phrase "unsubstituted alkoxy" refers to a hydroxyl group (-OH) in which the bond to the hydrogen atom is replaced by a bond to a carbon atom of an otherwise unsubstituted alkyl group as defined above.
[0069] The phrase "substituted alkoxy" refers to a hydroxyl group (-OH) in which the bond to the hydrogen atom is replaced by a bond to a carbon atom of an otherwise substituted alkyl group as defined above.
[0070] The phrase "unsubstituted heterocyclyloxy" refers to a hydroxyl group
(-OH) in which the bond to the hydrogen atom is replaced by a bond to a ring atom of an otherwise unsubstituted heterocyclyl group as defined above.
[0071] The phrase "substituted heterocyclyloxy" refers to a hydroxyl group (-
OH) in which the bond to the hydrogen atom is replaced by a bond to a ring atom of an otherwise substituted heterocyclyl group as defined above.
[0072] The phrase "unsubstituted heterocyclyloxyalkyl" refers to an unsubstituted alkyl group as defined above in which a carbon bond or hydrogen bond is replaced by a bond to an oxygen atom which is bonded to an unsubstituted heterocyclyl group as defined above.
[0073] The phrase "substituted heterocyclyloxyalkyl" refers to an unsubstituted heterocyclyloxyalkyl group as defined above in which a bond to a carbon or hydrogen group of the alkyl group of the heterocyclyloxyalkyl group is bonded to a non-carbon and non-hydrogen atom as described above with respect to substituted alkyl groups or in which the heterocyclyl group of the heterocyclyloxyalkyl group is a substituted heterocyclyl group as defined above.
[0074] The phrase "unsubstituted heterocyclylalkoxy" refers to an unsubstituted alkyl group as defined above in which a carbon bond or hydrogen bond is replaced by a bond to an oxygen atom which is bonded to the parent compound, and in which another carbon or hydrogen bond of the unsubstituted alkyl group is bonded to an unsubstituted heterocyclyl group as defined above.
[0075] The phrase "substituted heterocyclylalkoxy" refers to an unsubstituted heterocyclylalkoxy group as defined above in which a bond to a carbon or hydrogen group of the alkyl group of the heterocyclylalkoxy group is bonded to a non-carbon and non-hydrogen atom as described above with respect to substituted alkyl groups or in which the heterocyclyl group of the heterocyclylalkoxy group is a substituted heterocyclyl group as defined above. Further, a substituted heterocyclylalkoxy group also includes groups in which a carbon bond or a hydrogen bond to the alkyl moiety of the group may be substituted with one or more additional substituted and unsubstituted heterocycles. Examples include but are not limited to pyrid-2- ylmorpholin-4-ylmethyl and 2-pyrid-3-yl-2-morpholin-4-ylethyl.
[0076] The phrase "unsubstituted arylaminoalkyl" refers to an unsubstituted alkyl group as defined above in which a carbon bond or hydrogen bond is replaced by a bond to a nitrogen atom which is bonded to at least one unsubstituted aryl group as defined above.
[0077] The phrase "substituted arylaminoalkyl" refers to an unsubstituted arylaminoalkyl group as defined above except where either the alkyl group of the arylaminoalkyl group is a substituted alkyl group as defined above or the aryl group of the arylaminoalkyl group is a substituted aryl group except that the bonds to the nitrogen atom in all arylaminoalkyl groups does not by itself qualify all arylaminoalkyl groups as being substituted. However, substituted arylaminoalkyl groups does include groups in which the hydrogen bonded to the nitrogen atom of the group is replaced with a non-carbon and non-hydrogen atom. [0078] The phrase "unsubstituted heterocyclylaminoalkyl" refers to an unsubstituted alkyl group as defined above in which a carbon or hydrogen bond is replaced by a bond to a nitrogen atom which is bonded to at least one unsubstituted heterocyclyl group as defined above.
[0079] The phrase "substituted heterocyclylaminoalkyl" refers to unsubstituted heterocyclylaminoalkyl groups as defined above in which the heterocyclyl group is a substituted heterocyclyl group as defined above and/or the alkyl group is a substituted alkyl group as defined above. The bonds to the nitrogen atom in all heterocyclylaminoalkyl groups does not by itself qualify all heterocyclylaminoalkyl groups as being substituted. However, substituted heterocyclylaminoalkyl groups do include groups in which the hydrogen bonded to the nitrogen atom of the group is replaced with a non-carbon and non-hydrogen atom.
[0080] The phrase "unsubstituted alkylaminoalkoxy" refers to an unsubstituted alkyl group as defined above in which a carbon or hydrogen bond is replaced by a bond to an oxygen atom which is bonded to the parent compound and in which another carbon or hydrogen bond of the unsubstituted alkyl group is bonded to a nitrogen atom which is bonded to a hydrogen atom and an unsubstituted alkyl group as defined above.
[0081] The phrase "substituted alkylaminoalkoxy" refers to unsubstituted alkylaminoalkoxy groups as defined above in which a bond to a carbon or hydrogen atom of the alkyl group bonded to the oxygen atom which is bonded to the parent compound is replaced by one or more bonds to a non-carbon and non-hydrogen atoms as discussed above with respect to substituted alkyl groups and/or if the hydrogen bonded to the amino group is bonded to a non-carbon and non-hydrogen atom and/or if the alkyl group bonded to the nitrogen of the amine is bonded to a non-carbon and non-hydrogen atom as described above with respect to substituted alkyl groups. The presence of the amine and alkoxy functionality in all alkylaminoalkoxy groups does not by itself qualify all such groups as substituted alkylaminoalkoxy groups. [0082] The phrase "unsubstituted dialkylaminoalkoxy" refers to an unsubstituted alkyl group as defined above in which a carbon or hydrogen bond is replaced by a bond to an oxygen atom which is bonded to the parent compound and in which another carbon or hydrogen bond of the unsubstituted alkyl group is bonded to a nitrogen atom which is bonded to two other similar or different unsubstituted alkyl groups as defined above.
[0083] The phrase "substituted dialkylaminoalkoxy" refers to an unsubstituted dialkylaminoalkoxy group as defined above in which a bond to a carbon or hydrogen atom of the alkyl group bonded to the oxygen atom which is bonded to the parent compound is replaced by one or more bonds to a non-carbon and non-hydrogen atoms as discussed above with respect to substituted alkyl groups and/or if one or more of the alkyl groups bonded to the nitrogen of the amine is bonded to a non-carbon and non-hydrogen atom as described above with respect to substituted alkyl groups. The presence of the amine and alkoxy functionality in all dialkylaminoalkoxy groups does not by itself qualify all such groups as substituted dialkylaminoalkoxy groups.
[0084] The term "protected" with respect to hydroxyl groups, amine groups, and sulfhydryl groups refers to forms of these functionalities which are protected from undesirable reaction with a protecting group known to those skilled in the art such as those set forth in Protective Groups in Organic Synthesis, Greene, T.W.; Wuts, P. G. M., John Wiley & Sons, New York, NY, (3rd Edition, 1999) which can be added or removed using the procedures set forth therein. Examples of protected hydroxyl groups include, but are not limited to, silyl ethers such as those obtained by reaction of a hydroxyl group with a reagent such as, but not limited to, t-butyldimethyl- chlorosilane, trimefhylchlorosilane, triisopropylchlorosilane, triethylchlorosilane; substituted methyl and ethyl ethers such as, but not limited to methoxymethyl ether, methythiomethyl ether, benzyloxymethyl ether, /-butoxymefhyl ether, 2- methoxyethoxymethyl ether, tetrahydropyranyl ethers, 1-ethoxyethyl ether, allyl ether, benzyl ether; esters such as, but not limited to, benzoylformate, formate, acetate, trichloroacetate, and trifluoroacetate. Examples of protected amine groups include, but are not limited to, amides such as, formamide, acetamide, trifluoroacetamide, and benzamide; imides, such as phfhalimide, and dithiosuccinimide; and others. Examples of protected sulfhydryl groups include, but are not limited to, thioefhers such as S-benzyl thioether, and S-4-picolyl thioether; substituted S-methyl derivatives such as hemithio, dithio and aminothio acetals; and others.
[0085] A "pharmaceutically acceptable salt" includes a salt with an inorganic base, organic base, inorganic acid, organic acid, or basic or acidic amino acid. As salts of inorganic bases, the invention includes, for example, alkali metals such as sodium or potassium; alkaline earth metals such as calcium and magnesium or aluminum; and ammonia. As salts of organic bases, the invention includes, for example, trimethylamine, triethylamine, pyridine, picoline, ethanolamine, diethanolamine, and triethanolamine. As salts of inorganic acids, the instant invention includes, for example, hydrochloric acid, hydroboric acid, nitric acid, sulfuric acid, and phosphoric acid. As salts of organic acids, the instant invention includes, for example, formic acid, acetic acid, trifluoroacetic acid, fumaric acid, oxalic acid, tartaric acid, lactic acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, and p-toluenesulfonic acid. As salts of basic amino acids, the instant invention includes, for example, arginine, lysine and ornithine. Acidic amino acids include, for example, aspartic acid and glutamic acid.
[0086] In one aspect, the invention provides a method of modulating an inflammatory response and/or reducing cellular adhesion in a subject. Such methods include administering to the subject a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof. The inflammatory response is modulated in the subject and/or cellular adhesion is reduced in the subject after administration of the compound, the tautomer, the pharmaceutically acceptable salt of the compound, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof.
[0087] In one embodiment, the invention provides a method of treating a disorder related to inflammation in a human or animal subject. The method includes admimstering to the human or animal subject an effective amount of a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof. Inflammation and inflammatory responses may occur with various biological conditions. Examples of such biological conditions may include cancer, autoimmune diseases, asthma, allergies, eczema, microbial infections, traumatic injuries such as burns or cuts, lupus, arthritis, cardiovascular disease such as, but not limited to, strokes and ischemic injuries, respiratory bacterial and viral infections, and other conditions associated with inflammatory responses.
[0088] In another embodiment, the invention provides a method of treating a disorder related to cellular adhesion in a human or animal subject. The method includes administering to the human or animal subject an effective amount ofa compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof.
[0089] In another embodiment, the invention provides a method of decreasing cellular adhesion molecules such as ICAM, VCAM, E-selectin, MMP-2, or MMP-9 levels in a human or animal subject. The method includes administering to the human or animal subject a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof. The amount of the cellular adhesion molecule is typically reduced in the subject after administration.
[0090] In another embodiment, the invention provides a method of decreasing circulating ICAM, VCAM, E-selectin, MMP-2, or MMP-9 levels in a human or animal subject. The method includes administering to the human or animal subject a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof.
[0091] In another embodiment, the invention provides a method of decreasing circulating cell adhesion molecules in a human or animal subject. The method includes administering to the human or animal subject a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof.
[0092] In yet another embodiment, the invention provides a method of monitoring the progression ofa disease or treatment in a human or animal subject. The method includes administering to the human or animal subject a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof and measuring the amounts of a molecule such as ICAM, VCAM, E-selectin, MMP-2, or MMP-9 levels in the subject.
[0093] In another aspect, the invention provides a method of monitoring the progression of a disease or treatment in a human or animal subject. The method includes measuring the amount of at least one cell adhesion molecule in the subject after administration of a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof to the subject. In some embodiments, the cell adhesion molecule is selected from inducible cell adhesion molecule (ICAM), vascular cell adhesion molecule (VCAM), or endothelial leukocyte adhesion molecule (E-Selectin). Some such methods further include withdrawing a sample of blood from the subject and then measuring the amount of the at least one cell adhesion molecule in at least a portion of the sample.
[0094] In another aspect, the invention provides a method of identifying a subject in need of a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof. The method includes measuring the amount of at least one cell adhesion molecule in the subject before, during, or after administration of the compound of Structure I, the tautomer of the compound, the pharmaceutically acceptable salt of the compound, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof to the subject. In some embodiments, the cell adhesion molecule is selected from inducible cell adhesion molecule, vascular cell adhesion molecule, or endothelial leukocyte adhesion molecule. In some embodiments, the cell adhesion molecule is selected from inducible cell adhesion molecule (ICAM), vascular cell adhesion molecule (VCAM), or endothelial leukocyte adhesion molecule (E-Selectin). Some such methods further include withdrawing a sample of blood from the subject and then measuring the amount of the at least one cell adhesion molecule in at least a portion of the sample.
[0095] In some embodiments of any of the methods described herein, the subject is a cancer patient.
[0096] Structure I has the following formula:
Figure imgf000029_0001
wherein, R1, R2, R3, and R4 may be the same or different and are independently selected from the group consisting of H, Cl, Br, F, I, -CN, -NO2, -OH, -OR15 groups, -NR16R17 groups, substituted and unsubstituted amidinyl groups, substituted and unsubstituted guanidinyl groups, substituted and unsubstituted primary, secondary, and tertiary alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted alkenyl groups, substituted and unsubstituted alkynyl groups, substituted and unsubstituted heterocyclyl groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted heterocyclylalkyl groups, and -C(=O)R18 groups; R5, R6, R7, and R8 may be the same or different and are independently selected from the group consisting of H, Cl, Br, F, I, -NO2, -OH, -OR19 groups, -NR20R21 groups, -SH, -SR22 groups, -S(=O)R23 groups, -S(=O) R24 groups, -CN, substituted and unsubstituted amidinyl groups, substituted and unsubstituted guanidinyl groups, substituted and unsubstituted primary, secondary, and tertiary alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted alkenyl groups, substituted and unsubstituted alkynyl groups, substituted and unsubstituted heterocyclyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted heterocyclylalkyl groups, -C(=O)R25 groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted heterocyclylaminoalkyl groups, substituted and unsubstituted hydroxyalkyl groups, substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted aryloxyalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups; R12 is selected from the group consisting of H, substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, and substituted and unsubstituted heterocyclyl groups; R13 is selected from the group consisting of H, substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted heterocyclyl groups, -OH, alkoxy groups, aryloxy groups, -NH2, substituted and unsubstituted heterocyclylalkyl groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted alkylamino groups, substituted and unsubstituted arylamino groups, substituted and unsubstituted dialkylamino groups, substituted and unsubstituted diarylamino groups, substituted and unsubstituted (alkyl)(aryl)amino groups, -C(=O)H, -C(=O)-alkyl groups, -C(=O)-aryl groups, -C(=O)O-alkyl groups, -C(=O)O-aryl groups, -C(=O)NH2, -C(=O)NH(alkyl) groups, -C(=O)NH(aryl) groups, -C(=O)N(alkyl)2 groups, -C(=O)N(aryl)2 groups, -C(=O)N(alkyl)(aryl) groups, -C(=O)-heterocyclyl groups, -C(=O)-O-heterocyclyl groups, -C(=O)NH(heterocyclyl) groups, -C(=O)-N(heterocyclyl)2 groups, -C(=O)-N(alkyl)(heterocyclyl) groups, -C(=O)- N(aryl)(heterocyclyl) groups, substituted and unsubstituted heterocyclylaminoalkyl groups, substituted and unsubstituted hydroxyalkyl groups, substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted aryloxyalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups; R15 and R19 may be the same or different and are independently selected from the group consisting of substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted heterocyclyl groups, substituted and unsubstituted heterocyclylalkyl groups, -C(=O)H, -C(=O)-alkyl groups, -C(=O)-aryl groups, -C(=O)NH2, -C(=O)NH(alkyl) groups, -C(=O)NH(aryl) groups, -C(=O)N(alkyl)2 groups, -C(=O)N(aryl)2 groups, -C(=O)N(alkyl)(aryl) groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted heterocyclylaminoalkyl, substituted and unsubstituted diheterocyclylammoalkyl, substituted and unsubstituted (heterocyclyl)(alkyl)aminoalkyl, substituted and unsubstituted (heterocyclyl)(aryl)aminoalkyl, substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted hydroxyalkyl groups, substituted and unsubstituted aryloxyalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups; R16 and R20 may be the same or different and are independently selected from the group consisting of H, substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, and substituted and unsubstituted heterocyclyl groups; R17 and R21 may be the same or different and are independently selected from the group consisting of H, substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted heterocyclyl groups, -C(=O)H, -C(=O)-alkyl groups, -C(=O)-aryl groups,-C(=O)NH2, -C(=O)NH(alkyl) groups, -C(=O)NH(aryl) groups, -C(=O)N(alkyl)2 groups, -C(=O)N(aryl)2 groups, -C(=O)N(alkyl)(aryl) groups, -C(=O)O-alkyl groups, -C(=O)O-aryl groups, substituted and unsubstituted heterocyclylalkyl groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, -C(=O)-heterocyclyl groups, -C(=O)-O-heterocyclyl groups, -C(=O)NH(heterocyclyl) groups, -C(=O)-N(heterocyclyl)2 groups, -C(=O)-N(alkyl)(heterocyclyl) groups, -C(=O)-N(aryl)(heterocyclyl) groups, substituted and unsubstituted heterocyclylaminoalkyl groups, substituted and unsubstituted hydroxyalkyl groups, substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted aryloxyalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups; R18, R23, R24, and R25 may be the same or different and are independently selected from the group consisting of H, -NH2, -NH(alkyl) groups, -NH(aryl) groups, -N(alkyl)2 groups, -N(aryl)2 groups, -N(alkyl)(aryl) groups, -NH(heterocyclyl) groups, -N(heterocyclyl)(alkyl) groups, -N(heterocyclyl)(aryl) groups, -N(heterocyclyl)2 groups, substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, -OH, substituted and unsubstituted alkoxy groups, substituted and unsubstituted aryloxy groups, substituted and unsubstituted heterocyclyl groups, -NHOH, -N(alkyl)OH groups, -N(aryl)OH groups, -N(alkyl)O-alkyl groups, -N(aryl)O-alkyl groups, -N(alkyl)O-aryl groups, and -N(aryl)O-aryl groups; and R is selected from the group consisting of substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, and substituted and unsubstituted heterocyclyl groups.
[0097] In some embodiments of the pharmaceutically acceptable salts of the compounds or the tautomers of the compounds of Structure I, at least one of R5, R6, R7, or R8 is selected from the group consisting of substituted and unsubstituted amidinyl groups, substituted and unsubstituted guanidinyl groups, substituted and unsubstituted saturated heterocyclyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted heterocyclylalkyl groups, substituted and unsubstituted heterocyclylaminoalkyl groups, substituted and unsubstituted hydroxyalkyl groups, substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted aryloxyalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups; -OR19 groups wherein R19 is selected from the group consisting of substituted and unsubstituted aryl groups, substituted and unsubstituted heterocyclyl groups, substituted and unsubstituted heterocyclylalkyl groups, -C(=O)H, -C(=O)-aryl groups, -C(=O)NH , -C(=O)NH(alkyl) groups, -C(=O)NH(aryl) groups, -C(=O)N(alkyl)2 groups, -C(=O)N(aryl)2 groups, -C(=O)N(alkyl)(aryl) groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted heterocyclylaminoalkyl groups, substituted and unsubstituted diheterocyclylaminoalkyl groups, substituted and unsubstituted (heterocyclyl)(alkyl)aminoalkyl groups, substituted and unsubstituted (heterocyclyl)(aryl)aminoalkyl groups, substituted and unsubstituted hydroxyalkyl groups, substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted aryloxyalkyl groups, and substituted and unsubstituted 9H 1 0 heterocyclyloxyalkyl groups; -NR R groups wherein R is selected from the group consisting of substituted and unsubstituted heterocyclyl groups; -NR20R21 groups wherein R21 is selected from the group consisting of substituted and unsubstituted heterocyclyl groups, -C(=O)H, -C(=O)-aryl groups, -C(=O)NH2, -C(=O)NH(alkyl) groups, -C(=O)NH(aryι) groups, -C(=O)N(alkyl)2 groups, -C(=O)N(aryl)2 groups, -C(=O)N(alkyl)(aryl) groups, -C(=O)O-alkyl groups, -C(=O)O-aryl groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted heterocyclylaminoalkyl groups, substituted and unsubstituted hydroxyalkyl groups, substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted aryloxyalkyl groups, substituted and unsubstituted heterocyclylalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups; and -C(=O)R25 groups wherein R25 is selected from the group consisting of H, -NH2, -NH(alkyl) groups, -NH(aryl) groups, -N(alkyl)2 groups, -N(aryl)2 groups, -N(alkyι)(aryι) groups, -NH(heterocyclyl) groups, -N(heterocyclyl)(alkyl) groups, -N(heterocyclyl)(aryl) groups, -N(heterocyclyl)2 groups, substituted and unsubstituted aryl groups, substituted and unsubstituted aryloxy groups, and substituted and unsubstituted heterocyclyl groups.
[0098] In one embodiment, the invention relates to a pharmaceutically acceptable salt of 4-amino-5-fluoro-3-[6-(4-methylpiperazin-l -yl)-lH-benzimidazol- 2-yl]quinolin-2(lH)-one (Compound 1) or a tautomer thereof. In some such embodiments, the salt is selected from the group consisting of tartrate, malate, lactate, bis-acetate, citrate, mesylate, bismesylate and bishydrochloride.
[0099] In some specific embodiments, the compound of structure I is a lactate salt of 4-amino-5-fluoro-3-[6-(4-methylpiperazin- 1 -yl)- 1 H-benzimidazol-2- yl]quinolin-2(lH)-one or a tautomer thereof.
[0100] In some specific embodiments, the pharmaceutically acceptable salt of the compound of Structure I, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof is administered to the subject, and the salt is a lactate salt.
[0101] In some embodiments, at least one of R12 and R13 is H, and in other embodiments, both R12 and R13 are H.
[0102] In a some embodiments, R1 is selected from the group consisting of F,
Cl, substituted and unsubstituted alkoxy groups, substituted and unsubstituted heterocyclylalkoxy groups, substituted and unsubstituted heterocyclyl groups, substituted and unsubstituted alkylaminoalkoxy groups, substituted and unsubstituted arylaminoalkoxy groups, substituted and unsubstituted dialkylaminoalkoxy groups, substituted and unsubstituted diarylaminoalkoxy groups, and substituted and unsubstituted (alkyl)(aryl)aminoalkoxy groups.
[0103] In some embodiments, R1 is F and R2, R3, R3, R4, R5, and R8 are all H, and one of R6 or R7 is H.
[0104] In some other embodiments, at least one of R5, R6, R7, and R8 is a substituted or unsubstituted heterocyclyl group.
[0105] In still other embodiments, at least one of R5, R6, R7, and R8 is a substituted or unsubstituted heterocyclyl group comprising at least one O or N atom.
[0106] In yet other embodiments, at least one of R5, R6, R7, and R8 is a substituted or unsubstituted heterocyclyl group and the heterocyclyl group is selected from the group consisting of morpholine, piperazine, piperidine, pyrrolidine, thiomorpholine, homopiperazine, tetrahydrothiophene, tetrahydrofuran, and tetrahydropyran.
[0107] In yet other embodiments, at least one of R6 or R7 is a substituted or unsubstituted heterocyclyl group.
[0108] In yet other embodiments, at least one of R6 or R7 is a substituted or unsubstituted heterocyclyl group comprising at least one O or N atom.
[0109] In yet other embodiments, one of R6 or R7 is a substituted or unsubstituted heterocyclyl group and the heterocyclyl group is selected from the group consisting of morpholine, piperazine, piperidine, pyrrolidine, thiomorpholine, homopiperazine, tetrahydrothiophene, tetrahydrofuran, and tetrahydropyran.
[0110] In still other particular embodiments, one of R6 or R7 is selected from the group consisting of substituted and unsubstituted morpholine groups, and substituted and unsubstituted piperazine groups. In some such embodiments, one of R6 or R7 is a piperazine N-oxide or is an N-alkyl substituted piperazine.
[0111] In yet other embodiments, at least one of and in some embodiments one of R6 or R7 is selected from the group consisting of-NR20R21 groups wherein R20 is selected from the group consisting of substituted and unsubstituted heterocyclyl groups; and -NR20R21 groups wherein R21 is selected from the group consisting of substituted and unsubstituted heterocyclyl groups, groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted heterocyclylaminoalkyl groups, substituted and unsubstituted hydroxyalkyl groups, substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted aryloxyalkyl groups, substituted and unsubstituted heterocyclylalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups.
[0112] In yet another embodiment, R1 is selected from the group consisting of
H and F.
[0113] In yet another embodiment, the compounds and their corresponding salts and tautomers are provided in the following two tables below. The synthesis of these compounds is described in U.S. Patent No. 6,605,617, published U.S. Patent Application No. 2004/0092535, published U.S. Patent Application No. 2004/0220196 as are various kinase assay procedures. Each of these references is, therefore, hereby incorporated by reference in its entirety and for all purposes as if set forth in its entirety.
Table of Exemplary Compounds
Figure imgf000036_0001
Figure imgf000037_0001
Figure imgf000038_0001
Figure imgf000039_0001
Figure imgf000040_0001
Figure imgf000041_0001
Figure imgf000042_0001
Figure imgf000043_0001
Figure imgf000044_0001
Figure imgf000045_0001
Figure imgf000046_0001
Figure imgf000047_0001
Table of Additional Exemplary Compounds
Figure imgf000047_0002
Figure imgf000048_0001
Figure imgf000049_0001
Figure imgf000050_0001
Figure imgf000051_0001
Figure imgf000052_0001
Figure imgf000053_0001
Figure imgf000054_0001
Figure imgf000055_0001
Figure imgf000056_0001
Figure imgf000057_0001
Figure imgf000058_0001
Figure imgf000059_0001
Figure imgf000060_0001
Figure imgf000061_0001
Figure imgf000062_0001
Figure imgf000063_0001
Figure imgf000064_0001
Figure imgf000065_0001
Figure imgf000066_0001
Figure imgf000067_0001
Figure imgf000068_0001
Figure imgf000069_0001
Figure imgf000070_0001
Figure imgf000071_0001
Figure imgf000072_0001
Figure imgf000073_0001
Figure imgf000074_0001
Figure imgf000075_0001
Figure imgf000076_0001
Figure imgf000077_0001
Figure imgf000078_0001
Figure imgf000079_0001
Figure imgf000080_0001
Figure imgf000081_0001
Figure imgf000082_0001
Figure imgf000083_0001
Figure imgf000084_0001
Figure imgf000085_0001
Figure imgf000086_0001
Figure imgf000087_0001
Figure imgf000088_0003
[0114] In yet another embodiment, the compound of Structure I is a compound of Structure II, where Structure II has the following formula:
Figure imgf000088_0001
II wherein,
A is a group having one of the following Structures:
Figure imgf000088_0002
wherein,
Ra is selected from H or straight or branched chain alkyl groups having from 1 to 6 carbon atoms.
[0115] In some embodiments, where the compound of Structure I is a compound of Structure II, Ra is a methyl group, and the compound of Structure II is a compound of Structure II A
Figure imgf000089_0001
IIA
[0116] In some specific embodiments, the pharmaceutically acceptable salt of the compound of Structure IIA, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof is administered to the subject, and the salt is a lactate salt.
[0117] In some embodiments, where the compound of Structure I is a compound of Structure II, Ra is a H, and the compound of Structure II is a compound of Structure IIB
Figure imgf000089_0002
IIB
[0118] In some embodiments, where the compound of Structure I is a compound of Structure II, Ra is a methyl group, and the compound of Structure II is a compound of Structure IIC
Figure imgf000090_0001
IIC
[0119] The compounds of any of the embodiments may be used to prepare medicaments or pharmaceutical formulations for use in any of the methods of the invention.
[0120] Pharmaceutical formulations for use with the invention may include any of the compounds, tautomers, or salts of any of the embodiments described above in combination with a pharmaceutically acceptable carrier such as those described herein.
[0121] The instant invention also provides for compositions which may be prepared by mixing one or more compounds of the instant invention, or pharmaceutically acceptable salts tautomers thereof, or mixtures thereof with pharmaceutically acceptable carriers, excipients, binders, diluents or the like to treat or ameliorate disorders related to metastacized tumors. The compositions of the inventions may be used to create formulations for use in any of the methods of the invention. Such compositions can be in the form of, for example, granules, powders, tablets, capsules, syrup, suppositories, injections, emulsions, elixirs, suspensions or solutions. The instant compositions can be formulated for various routes of administration, for example, by oral administration, by nasal administration, by rectal administration, subcutaneous injection, intravenous injection, intramuscular injections, or intraperitoneal injection. The following dosage forms are given by way of example and should not be construed as limiting the instant invention.
[0122] For oral, buccal, and sublingual administration, powders, suspensions, granules, tablets, pills, capsules, gelcaps, and caplets are acceptable as solid dosage forms. These can be prepared, for example, by mixing one or more compounds of the instant invention, pharmaceutically acceptable salts, tautomers, or mixtures thereof, with at least one additive such as a starch or other additive. Suitable additives are sucrose, lactose, cellulose sugar, mannitol, maltitol, dextran, starch, agar, alginates, chitins, chitosans, pectins, tragacanth gum, gum arabic, gelatins, collagens, casein, albumin, synthetic or semi-synthetic polymers or glycerides. Optionally, oral dosage forms can contain other ingredients to aid in administration, such as an inactive diluent, or lubricants such as magnesium stearate, or preservatives such as paraben or sorbic acid, or anti-oxidants such as ascorbic acid, tocopherol or cysteine, a disintegrating agent, binders, thickeners, buffers, sweeteners, flavoring agents or perfuming agents. Tablets and pills may be further treated with suitable coating materials known in the art.
[0123] Liquid dosage forms for oral administration may be in the form of pharmaceutically acceptable emulsions, syrups, elixirs, suspensions, and solutions, which may contain an inactive diluent, such as water. Pharmaceutical formulations and medicaments may be prepared as liquid suspensions or solutions using a sterile liquid, such as, but not limited to, an oil, water, an alcohol, and combinations of these. Pharmaceutically suitable surfactants, suspending agents, emulsifying agents, may be added for oral or parenteral administration.
[0124] As noted above, suspensions may include oils. Such oil include, but are not limited to, peanut oil, sesame oil, cottonseed oil, corn oil and olive oil. Suspension preparation may also contain esters of fatty acids such as ethyl oleate, isopropyl myristate, fatty acid glycerides and acetylated fatty acid glycerides. Suspension formulations may include alcohols, such as, but not limited to, ethanol, isopropyl alcohol, hexadecyl alcohol, glycerol and propylene glycol. Ethers, such as but not limited to, poly(ethyleneglycol), petroleum hydrocarbons such as mineral oil and petrolatum; and water may also be used in suspension formulations.
[0125] For nasal administration, the pharmaceutical formulations and medicaments may be a spray or aerosol containing an appropriate solvent(s) and optionally other compounds such as, but not limited to, stabilizers, antimicrobial agents, antioxidants, pH modifiers, surfactants, bioavailability modifiers and combinations of these. A propellant for an aerosol formulation may include compressed air, nitrogen, carbon dioxide, or a hydrocarbon based low boiling solvent.
[0126] Injectable dosage forms generally include aqueous suspensions or oil suspensions which may be prepared using a suitable dispersant or wetting agent and a suspending agent. Injectable forms may be in solution phase or in the form of a suspension, which is prepared with a solvent or diluent. Acceptable solvents or vehicles include sterilized water, Ringer's solution, or an isotonic aqueous saline solution. Alternatively, sterile oils may be employed as solvents or suspending agents. Preferably, the oil or fatty acid is non- volatile, including natural or synthetic oils, fatty acids, mono-, di- or tri-glycerides.
[0127] For injection, the pharmaceutical formulation and/or medicament may be a powder suitable for reconstitution with an appropriate solution as described above. Examples of these include, but are not limited to, freeze dried, rotary dried or spray dried powders, amorphous powders, granules, precipitates, or particulates. For injection, the formulations may optionally contain stabilizers, pH modifiers, surfactants, bioavailability modifiers and combinations of these.
[0128] For rectal administration, the pharmaceutical formulations and medicaments may be in the form of a suppository, an ointment, an enema, a tablet or a cream for release of compound in the intestines, sigmoid flexure and/or rectum. Rectal suppositories are prepared by mixing one or more compounds of the instant invention, or pharmaceutically acceptable salts or tautomers of the compound, with acceptable vehicles, for example, cocoa butter or polyethylene glycol, which is present in a solid phase at normal storing temperatures, and present in a liquid phase at those temperatures suitable to release a drug inside the body, such as in the rectum. Oils may also be employed in the preparation of formulations of the soft gelatin type and suppositories. Water, saline, aqueous dextrose and related sugar solutions, and glycerols may be employed in the preparation of suspension formulations which may also contain suspending agents such as pectins, carbomers, methyl cellulose, hydroxypropyl cellulose or carboxymethyl cellulose, as well as buffers and preservatives.
[0129] Besides those representative dosage forms described above, pharmaceutically acceptable excipients and carriers are generally known to those skilled in the art and are thus included in the instant invention. Such excipients and carriers are described, for example, in "Remingtons Pharmaceutical Sciences" Mack Pub. Co., New Jersey (1991), which is incorporated herein by reference in its entirety for all purposes as if fully set forth herein.
[0130] The formulations of the invention may be designed to be short-acting, fast-releasing, long-acting, and sustained-releasing as described below. Thus, the pharmaceutical formulations may also be formulated for controlled release or for slow release.
[0131] The instant compositions may also comprise, for example, micelles or liposomes, or some other encapsulated form, or may be administered in an extended release form to provide a prolonged storage and/or delivery effect. Therefore, the pharmaceutical formulations and medicaments may be compressed into pellets or cylinders and implanted intramuscularly or subcutaneously as depot injections or as implants such as stents. Such implants may employ known inert materials such as silicones and biodegradable polymers.
[0132] Specific dosages may be adjusted depending on conditions of disease, the age, body weight, general health conditions, sex, and diet of the subject, dose intervals, administration routes, excretion rate, and combinations of drugs. Any of the above dosage forms containing effective amounts are well within the bounds of routine experimentation and therefore, well within the scope of the instant invention.
[0133] A therapeutically effective dose may vary depending upon the route of administration and dosage form. The preferred compound or compounds of the instant invention is a formulation that exhibits a high therapeutic index. The therapeutic index is the dose ratio between toxic and therapeutic effects which can be expressed as the ratio between LD50 and ED50. The LD50 is the dose lethal to 50% of the population and the ED50 is the dose therapeutically effective in 50% of the population. The LD5o and ED5o are determined by standard pharmaceutical procedures in animal cell cultures or experimental animals.
[0134] Pharmaceutical formulations and medicaments according to the invention include the compound of Structure I or the tautomers, salts, or mixtures thereof in combination with a pharmaceutically acceptable carrier. Thus, the compounds of the invention may be used to prepare medicaments and pharmaceutical formulations. Such medicaments and pharmaceutical formulations may be used in any of the methods of treatment described herein.
[0135] The compounds and formulations of the present invention are particularly suitable for use in combination therapy. Kinase inhibitors for use as anticancer agents in conjunction with the methods or compositions of the present invention include inhibitors of Epidermal Growth Factor Receptor (EGFR) kinases such as small molecule quinazolines, for example gefitinib (US 5457105, US 5616582, and US 5770599), ZD-6474 (WO 01/32651), erlotinib (Tarceva®, US 5,747,498 and WO 96/30347), and lapatinib (US 6,727,256 and WO 02/02552). Kinase inhibitors for use as anticancer agents in conjunction with the methods of compositions of the present invention also include inibitors of Vascular Endothelial Growth Factor Receptor (VEGFR) kinase inhibitor such as, but not limited to, SU- 11248 (WO 01/60814), SU 5416 (US 5,883,113 and WO 99/61422), SU 6668 (US 5,883,113 and WO 99/61422), CHIR-258 (US 6,605,617 and US 6,774,237), vatalanib or PTK-787 (US 6,258,812), VEGF-Trap (WO 02/57423), B43-Genistein (WO-09606116), fenre inide (retinoic acid p-hydroxyphenylamine) (US 4,323,581), IM-862 (WO 02/62826), bevacizumab or Avastin® (WO 94/10202), KRN-951, 3-[5- (methylsulfonylpiperadine methyl)-indolyl]-quinolone, AG-13736 and AG-13925, pyrrolo[2,l-f][l,2,4]triazines, ZK-304709, Veglin®, VMDA-3601, EG-004, CEP-701 (US 5,621,100), and Cand5 (WO 04/09769).
[0136] The compounds of the invention may be used to treat a variety of subjects. Suitable subjects include animals such as mammals and humans. Suitable mammals include, but are not limited to, primates such as, but not limited to lemurs, apes, and monkeys; rodents such as rats, mice, and guinea pigs; rabbits and hares; cows; horses; pigs; goats; sheep; marsupials; and carnivores such as felines, canines, and ursines. In some embodiments, the subject or patient is a human. In other embodiments, the subject or patient is a rodent such as a mouse or a rat. In some embodiments, the subject or patient is an animal other than a human and in some such embodiments, the subject or patient is a mammal other than a human.
Purification and Characterization of Compounds
[0137] Compounds of the present invention were characterized by high performance liquid chromatography (HPLC) using a Waters Millenium chromatography system with a 2690 Separation Module (Milford, Massachusetts). The analytical columns were Alltima C-18 reversed phase, 4.6 x 250 mm from Alltech (Deerfield, Illinois). A gradient elution was used, typically starting with 5% acetonitrile/95% water and progressing to 100% acetonitrile over a period of 40 minutes. All solvents contained 0.1%) trifluoroacetic acid (TFA). Compounds were detected by ultraviolet light (UV) absorption at either 220 or 254 nm. HPLC solvents were from Burdick and Jackson (Muskegan, Michigan), or Fisher Scientific (Pittsburg, Pennsylvania). In some instances, purity was assessed by thin layer chromatography (TLC) using glass or plastic backed silica gel plates, such as, for example, Baker-Flex Silica Gel 1B2-F flexible sheets. TLC results were readily detected visually under ultraviolet light, or by employing well known iodine vapor and other various staining techniques.
[0138] Mass spectrometric analysis was performed on one of two LCMS instruments: a Waters System (Alliance HT HPLC and a Micromass ZQ mass spectrometer; Column: Eclipse XDB-C18, 2.1 x 50 mm; Solvent system: 5-95% acetonitrile in water with 0.05% TFA; Flow rate 0.8 mL/minute; Molecular weight range 150-850; Cone Voltage 20 V; Column temperature 40°C) or a Hewlett Packard System (Series 1100 HPLC; Column: Eclipse XDB-C18, 2.1 x 50 mm; Solvent system: 1-95%) acetonitrile in water with 0.05%> TFA; Flow rate 0.4 mL/minute; Molecular weight range 150-850; Cone Voltage 50 V; Column temperature 30°C). All masses are reported as those of the protonated parent ions. [0139] GCMS analysis was performed on a Hewlet Packard instrument
(HP6890 Series gas chromatograph with a Mass Selective Detector 5973; Injector volume: 1 μL; Initial column temperature: 50°C; Final column temperature: 250°C; Ramp time: 20 minutes; Gas flow rate: 1 mL/minute; Column: 5%> Phenyl Methyl Siloxane, Model #HP 190915-443, Dimensions: 30.0 m x 25 μm x 0.25 μm).
[0140] Preparative separations were carried out using either a Flash 40 chromatography system and KP-Sil, 60A (Biotage, Charlottesville, Virginia), or by HPLC using a C-18 reversed phase column. Typical solvents employed for the Flash 40 Biotage system were dichloromethane, methanol, ethyl acetate, hexane and triethyl amine. Typical solvents employed for the reverse phase HPLC were varying concentrations of acetonitrile and water with 0.1% trifluoroacetic acid.
Synthesis of 4-Amino-5-fluoro-3-[6-(4-methylpiperazin-l-yl)-lH-benzimidazol-2-
Figure imgf000096_0001
A. Synthesis of 5-(4-Methyl-piperazin-l-yl)-2-nitroaniline Procedure A
Figure imgf000096_0002
[0141] 5-Chloro-2-nitroaniline (500 g, 2.898 mol) and 1-methyl piperazine
(871 g, 8.693 mol) were placed in a 2000 mL flask fitted with a condenser and purged with N2. The flask was placed in an oil bath at 100°C and heated until the 5-chloro-2- nitroaniline was completely reacted (typically overnight) as determined by HPLC. After HPLC confirmed the disappearance of the 5-chloro-2-nitroaniline, the reaction mixture was poured directly (still warm) into 2500 mL of room temperature water with mechanical stirring. The resulting mixture was stirred until it reached room temperature and then it was filtered. The yellow solid thus obtained was added to 1000 mL of water and stirred for 30 minutes. The resulting mixture was filtered, and the resulting solid was washed with TBME (500 mL, 2X) and then was dried under vacuum for one hour using a rubber dam. The resulting solid was transferred to a drying tray and dried in a vacuum oven at 50°C to a constant weight to yield 670 g (97.8%) of the title compound as a yellow powder.
Procedure B
[0142] 5-Chloro-2-nitroaniline (308.2 g, 1.79 mol) was added to a 4-neck
5000 mL round bottom flask fitted with an overhead stirrer, condenser, gas inlet, addition funnel, and thermometer probe. The flask was then purged with N2. 1- Methylpiperazine (758.1 g, 840 mL, 7.57 mol) and 200 proof ethanol (508 mL) were added to the reaction flask with stirring. The flask was again purged with N2, and the reaction was maintained under N2. The flask was heated in a heating mantle to an internal temperature of 97°C (+/- 5°C) and maintained at that temperature until the reaction was complete (typically about 40 hours) as determined by HPLC. After the reaction was complete, heating was discontinued and the reaction was cooled to an internal temperature of about 20°C to 25°C with stirring, and the reaction was stirred for 2 to 3 hours. Seed crystals (0.20 g, 0.85 mmol) of 5-(4-methyl-piperazin-l-yl)-2- nitroaniline were added to the reaction mixture unless precipitation had already occurred. Water (2,450 mL) was added to the stirred reaction mixture over a period of about one hour while the internal temperature was maintained at a temperature ranging from about 20°C to 30°C. After the addition of water was complete, the resulting mixture was stirred for about one hour at a temperature of 20°C to 30°C. The resulting mixture was then filtered, and the flask and filter cake were washed with water (3 x 2.56 L). The golden yellow solid product was dried to a constant weight of 416 g (98.6%) yield) under vacuum at about 50°C in a vacuum oven.
Procedure C
[0143] 5-Chloro-2-nitroaniline (401 g, 2.32 mol) was added to a 4-neck 12 L round bottom flask fitted with an overhead stirrer, condenser, gas inlet, addition funnel, and thermometer probe. The flask was then purged with N2. 1- Me hylpiperazine (977 g, 1.08 L, 9.75 mol) and 100% ethanol (650 mL) were added to the reaction flask with stirring. The flask was again purged with N , and the reaction was maintained under N2. The flask was heated in a heating mantle to an internal temperature of 97°C (+/- 5°C) and maintained at that temperature until the reaction was complete (typically about 40 hours) as determined by HPLC. After the reaction was complete, heating was discontinued and the reaction was cooled to an internal temperature of about 80°C with stirring, and water (3.15 L) was added to the mixture via an addition funnel over the period of 1 hour while the internal temperature was maintained at 82°C (+/- 3°C). After water addition was complete, heating was discontinued and the reaction mixture was allowed to cool over a period of no less than 4 hours to an internal temperature of 20-25°C. The reaction mixture was then stirred for an additional hour at an internal temperature of 20-30°C. The resulting mixture was then filtered, and the flask and filter cake were washed with water (l x l L), 50% ethanol (1 x IL), and 95% ethanol (1 x IL). The golden yellow solid product was placed in a drying pan and dried to a constant weight of 546 g (99% yield) under vacuum at about 50°C in a vacuum oven.
B. Synthesis of [6-(4-Methyl-piperazin-l-yl)-lH-benzimidazol-2-yl]-acetic acid ethyl ester
Procedure A
Figure imgf000099_0001
[0144] A 5000 mL, 4-neck flask was fitted with a stirrer, thermometer, condenser, and gas inlet/outlet. The equipped flask was charged with 265.7 g (1.12 mol. 1.0 eq) of 5-(4-methyl-piperazin-l-yl)-2-nitroaniline and 2125 mL of 200 proof EtOH. The resulting solution was purged with N2 for 15 minutes. Next, 20.0 g of 5% Pd/C (50% H O w/w) was added. The reaction was vigorously stirred at 40-50°C (internal temperature) while H2 was bubbled through the mixture. The reaction was monitored hourly for the disappearance of 5-(4-methyl-piperazin-l-yl)-2-nitroaniline by HPLC. The typical reaction time was 6 hours.
[0145] After all the 5-(4-methyl-piperazin-l-yl)-2-nitroaniline had disappeared from the reaction, the solution was purged with N2 for 15 minutes. Next, 440.0 g (2.25 mol) of ethyl 3-ethoxy-3-iminopropanoate hydrochloride was added as a solid. The reaction was stirred at 40-50°C (internal temperature) until the reaction was complete. The reaction was monitored by following the disappearance of the diamino compound by HPLC. The typical reaction time was 1-2 hours. After the reaction was complete, it was cooled to room temperature and filtered through a pad of Celite filtering material. The Celite filtering material was washed with absolute EtOH (2 x 250 mL), and the filtrate was concentrated under reduced pressure providing a thick brown orange oil. The resulting oil was taken up in 850 mL of a 0.37% HCl solution. Solid NaOH (25 g) was then added in one portion, and a precipitate formed. The resulting mixture was stirred for 1 hour and then filtered. The solid was washed with H2O (2 x 400 mL) and dried at 50°C in a vacuum oven providing 251.7 g (74.1%) of [6-(4-methyl-piperazin-l-yl)-lH-benzoimidazol-2-yl]- acetic acid ethyl ester as a pale yellow powder.
Procedure B
[0146] A 5000 mL, 4-neck jacketed flask was fitted with a mechanical stirrer, condenser, temperature probe, gas inlet, and oil bubbler. The equipped flask was charged with 300 g (1.27 mol) of 5-(4-methyl-piperazin-l-yl)-2-nitroaniline and 2400 mL of 200 proof EtOH (the reaction may be and has been conducted with 95% ethanol and it is not necessary to use 200 proof ethanol for this reaction). The resulting solution was stirred and purged with N2 for 15 minutes. Next, 22.7 g of 5% Pd/C (50% H2O w/w) was added to the reaction flask. The reaction vessel was purged with N2 for 15 minutes. After purging with N2, the reaction vessel was purged with H2 by maintaining a slow, but constant flow of H2 through the flask. The reaction was stirred at 45-55°C (internal temperature) while H2 was bubbled through the mixture until the 5-(4-methyl-piperazin-l-yl)-2-nitroaniline was completely consumed as determined by HPLC. The typical reaction time was 6 hours.
[0147] After all the 5-(4-methyl-piperazin-l-yl)-2-nitroaniline had disappeared from the reaction, the solution was purged with N2 for 15 minutes. The diamine intermediate is air sensitive so care was taken to avoid exposure to air. 500 g (2.56 mol) of ethyl 3-ethoxy-3-iminopropanoate hydrochloride was added to the reaction mixture over a period of about 30 minutes. The reaction was stirred at 45- 55°C (internal temperature) under N2 until the diamine was completely consumed as determined by HPLC. The typical reaction time was about 2 hours. After the reaction was complete, the reaction was filtered while warm through a pad of Celite. The reaction flask and Celite were then washed with 200 proof EtOH (3 x 285 mL). The filtrates were combined in a 5000 mL flask, and about 3300 mL of ethanol was removed under vacuum producing an orange oil. Water (530 mL) and then IM HCL (350 mL) were added to the resulting oil, and the resulting mixture was stirred. The resulting solution was vigorously stirred while 30% NaOH (200 mL) was added over a period of about 20 minutes maintaining the internal temperature at about 25-30°C while the pH was brought to between 9 and 10. The resulting suspension was stirred for about 4 hours while maintaining the internal temperature at about 20-25°C. The resulting mixture was filtered, and the filter cake was washed with H2O (3 x 300 mL). The collected solid was dried to a constant weight at 50°C under vacuum in a vacuum oven providing 345.9 g (90.1%) of [6-(4-methyl-piperazin-l-yl)-lH-benzoimidazol-2- yl] -acetic acid ethyl ester as a pale yellow powder. In an alternative work up procedure, the filtrates were combined and the ethanol was removed under vacuum until at least about 90% had been removed. Water at a neutral pH was then added to the resulting oil, and the solution was cooled to about 0°C. An aqueous 20% NaOH solution was then added slowly with rapid stirring to bring the pH up to 9.2 (read with pH meter). The resulting mixture was then filtered and dried as described above. The alternative work up procedure provided the light tan to light yellow product in yields as high as 97%.
Method for Reducing Water Content of [6-(4-Methyl-piperazin-l-yl)-lH- benzoimidazol-2-yl] -acetic acid ethyl ester
[0148] [6-(4-Methyl-piperazin-l-yl)-lH-benzimidazol-2-yl] -acetic acid ethyl ester (120.7 grams) that had been previously worked up and dried to a water content of about 8-9% H2O was placed in a 2000 mL round bottom flask and dissolved in absolute ethanol (500 mL). The amber solution was concentrated to a thick oil using a rotary evaporator with heating until all solvent was removed. The procedure was repeated two more times. The thick oil thus obtained was left in the flask and placed in a vacuum oven heated at 50°C overnight. Karl Fisher analysis results indicated a water content of 5.25%. The lowered water content obtained by this method provided increased yields in the procedure of the following Example. Other solvents such as toluene and THF may be used in place of the ethanol for this drying process. C. Synthesis of 4-Amino-5-fluoro-3-[6-(4-methyl-piperazin-l-yl)-lH- benzimidazol-2-yl]-lH-quinolin-2-one
Procedure A
Figure imgf000102_0001
[0149] [6-(4-Methyl-piperazin-l-yl)-lH-benzimidazol-2-yl]-acetic acid ethyl ester (250 g, 820 mmol) (dried with ethanol as described above) was dissolved in THF (3800 mL) in a 5000 mL flask fitted with a condenser, mechanical stirrer, temperature probe, and purged with argon. 2-Amino-6-fluoro-benzonitrile (95.3 g, 700 mmol) was added to the solution, and the internal temperature was raised to 40°C. When all the solids had dissolved and the solution temperature had reached 40°C, solid KHMDS (376.2 g, 1890 mmol) was added over a period of 5 minutes. When addition of the potassium base was complete, a heterogeneous yellow solution was obtained, and the internal temperature had risen to 62°C. After a period of 60 minutes, the internal temperature decreased back to 40°C, and the reaction was determined to be complete by HPLC (no starting material or uncyclized intermediate was present). The thick reaction mixture was then quenched by pouring it into H2O (6000 mL) and stirring the resulting mixture until it had reached room temperature. The mixture was then filtered, and the filter pad was washed with water (1000 mL 2X). The bright yellow solid was placed in a drying tray and dried in a vacuum oven at 50°C overnight providing 155.3 g (47.9%) of the desired 4-amino-5-fTuoro-3-[6-(4- mefhyl-piperazin- 1 -yl)- 1 H-benzimidazol-2-yl] - 1 H-quinolin-2-one .
Procedure B
[0150] A 5000 mL 4-neck jacketed flask was equipped with a distillation apparatus, a temperature probe, a N2 gas inlet, an addition funnel, and a mechanical stirrer. [6-(4-Methyl-piperazin-l-yl)-lH-benzimidazol-2-yl]-acetic acid ethyl ester (173.0 g, 570 mmol) was charged into the reactor, and the reactor was purged with N2 for 15 minutes. Dry THF (2600 mL) was then charged into the flask with stirring. After all the solid had dissolved, solvent was removed by distillation (vacuum or atmospheric (the higher temperature helps to remove the water) using heat as necessary. After 1000 mL of solvent had been removed, distillation was stopped and the reaction was purged with N2. 1000 mL of dry THF was then added to the reaction vessel, and when all solid was dissolved, distillation (vacuum or atmospheric) was again conducted until another 1000 mL of solvent had been removed. This process of adding dry THF and solvent removal was repeated at least 4 times (on the 4th distillation, 60% of the solvent is removed instead of just 40% as in the first 3 distillations) after which a 1 mL sample was removed for Karl Fischer analysis to determine water content. If the analysis showed that the sample contained less than 0.20% water, then reaction was continued as described in the next paragraph. However, if the analysis showed more than 0.20% water, then the drying process described above was continued until a water content of less than 0.20% was achieved.
[0151] After a water content of less than or about 0.20% was achieved using the procedure described in the previous paragraph, the distillation apparatus was replaced with a reflux condenser, and the reaction was charged with 2-amino-6- fluoro-benzonitrile (66.2 g, 470 mmol) ( in some procedures 0.95 equivalents is used). The reaction was then heated to an internal temperature of 38-42°C. When the internal temperature had reached 38-42°C, KHMDS solution (1313 g, 1.32 mol, 20% KHMDS in THF) was added to the reaction via the additional funnel over a period of 5 minutes maintaining the internal temperature at about 38-50°C during the addition. When addition of the potassium base was complete, the reaction was stirred for 3.5 to 4.5 hours (in some examples it was stirred for 30 to 60 minutes and the reaction may be complete within that time) while maintaining the internal temperature at from 38- 42°C. A sample of the reaction was then removed and analyzed by HPLC. If the reaction was not complete, additional KHMDS solution was added to the flask over a period of 5 minutes and the reaction was stirred at 38-42°C for 45-60 minutes (the amount of KHMDS solution added was determined by the following: If the IPC ratio is < 3.50, then 125 mL was added; if 10.0 > IPC ratio > 3.50, then 56 mL was added; if 20.0 > IPC ratio > 10, then 30 mL was added. The IPC ratio is equal to the area corresponding to 4-amino-5-fluoro-3-[6-(4-methyl-piperazin-l-yl)-lH-benzimidazol- 2-yl]-lH-quinolin-2-one) divided by the area corresponding to the uncyclized intermediate). Once the reaction was complete (IPC ratio > 20), the reactor was cooled to an internal temperature of 25-30°C, and water (350 mL) was charged into the reactor over a period of 15 minutes while maintaining the internal temperature at 25-35°C (in one alternative, the reaction is conducted at 40°C and water is added within 5 minutes. The quicker quench reduces the amount of impurity that forms over time). The reflux condenser was then replaced with a distillation apparatus and solvent was removed by distillation (vacuum or atmospheric) using heat as required. After 1500 mL of solvent had been removed, distillation was discontinued and the reaction was purged with N2. Water (1660 mL) was then added to the reaction flask while maintaining the internal temperature at 20-30°C. The reaction mixture was then stirred at 20-30°C for 30 minutes before cooling it to an internal temperature of 5- 10°C and then stirring for 1 hour. The resulting suspension was filtered, and the flask and filter cake were washed with water (3 x 650 mL). The solid thus obtained was dried to a constant weight under vacuum at 50°C in a vacuum oven to provide 103.9 g (42.6% yield) of 4-amino-5-fluoro-3-[6-(4-methyl-piperazin-l-yl)-lH-benzimidazol- 2-yl]-lH-quinolin-2-one as a yellow powder.
Procedure C
Figure imgf000104_0001
[0152] [6-(4-Methyl-piperazin- 1 -yl)- 1 H-benzimidazol-2-yl] -acetic acid ethyl ester (608 g, 2.01 mol) (dried) and 2-amino-6-fluoro-benzonitrile (274 g, 2.01 mol) were charged into a 4-neck 12 L flask seated on a heating mantle and fitted with a condenser, mechanical stirrer, gas inlet, and temperature probe. The reaction vessel was purged with N2, and toluene (7.7 L) was charged into the reaction mixture while it was stirred. The reaction vessel was again purged with N2 and maintained under N2. The internal temperature of the mixture was raised until a temperature of 63°C (+/- 3°C) was achieved. The internal temperature of the mixture was maintained at 63°C (+/- 3°C) while approximately 2.6 L of toluene was distilled from the flask under reduced pressure (380 +/- 10 torr, distilling head t = 40°C (+/- 10°C) (Karl Fischer analysis was used to check the water content in the mixture. If the water content was greater than 0.03%, then another 2.6 L of toluene was added and distillation was repeated. This process was repeated until a water content of less than 0.03% was achieved). After a water content of less than 0.03% was reached, heating was discontinued, and the reaction was cooled under N2 to an internal temperature of 17-19°C. Potassium t-butoxide in THF (20% in THF; 3.39 kg, 6.04 moles potassium t-butoxide) was then added to the reaction under N2 at a rate such that the internal temperature of the reaction was kept below 20°C. After addition of the potassium t- butoxide was complete, the reaction was stirred at an internal temperature of less than 20°C for 30 minutes. The temperature was then raised to 25°C, and the reaction was stirred for at least 1 hour. The temperature was then raised to 30°C, and the reaction was stirred for at least 30 minutes. The reaction was then monitored for completion using HPLC to check for consumption of the starting materials (typically in 2-3 hours, both starting materials were consumed (less than 0.5% by area % HPLC)). If the reaction was not complete after 2 hours, another 0.05 equivalents of potassium t- butoxide was added at a time, and the process was completed until HPLC showed that the reaction was complete. After the reaction was complete, 650 mL of water was added to the stirred reaction mixture. The reaction was then warmed to an internal temperature of 50°C and the THF was distilled away (about 3 L by volume) under reduced pressure from the reaction mixture. Water (2.6 L) was then added dropwise to the reaction mixture using an addition funnel. The mixture was then cooled to room temperature and stirred for at least 1 hour. The mixture was then filtered, and the filter cake was washed with water (1.2 L), with 70% ethanol (1.2 L), and with 95%) ethanol (1.2 L). The bright yellow solid was placed in a drying tray and dried in a vacuum oven at 50°C until a constant weight was obtained providing 674 g (85.4%) of the desired 4-amino-5-fluoro-3-[6-(4-methyl-piperazin- 1 -yl)- lH-benzimidazol-2- yl] - 1 H-quinolin-2-one . Purification of 4-Amino-5-fluoro-3-[6-(4-methyl-piperazin-l-yl)-lH- benzimidazoI-2-yl]-lH-quinolin-2-one
[0153] A 3000 mL 4-neck flask equipped with a condenser, temperature probe, N2 gas inlet, and mechanical stirrer was placed in a heating mantle. The flask was then charged with 4-amino-5-fluoro-3-[6-(4-methyl-piperazin-l-yl)-lH- benzimidazol-2-yl]-lH-quinolin-2-one (101.0 g, 0.26 mol), and the yellow solid was suspended in 95% ethanol (1000 mL) and stirred. In some cases an 8:1 solvent ratio is used. The suspension was then heated to a gentle reflux (temperature of about
76°C) with stirring over a period of about 1 hour. The reaction was then stirred for
45-75 minutes while refluxed. At this point, the heat was removed from the flask and the suspension was allowed to cool to a temperature of 25-30°C. The suspension was then filtered, and the filter pad was washed with water (2 x 500 mL). The yellow solid was then placed in a drying tray and dried in a vacuum oven at 50°C until a constant weight was obtained (typically 16 hours) to obtain 97.2 g (96.2%>) of the purified product as a yellow powder.
D. Preparation of Lactic Acid Salt of 4-Amino-5-fluoro-3-[6-(4-methyl- piperazin-l-yl)-lH-benzimidazol-2-yl]-lH-quinolin-2-one
Figure imgf000106_0001
D,L-Lactic Acid EtOH, H20
Figure imgf000106_0002
[0154] A 3000 mL 4-necked jacketed flask was fitted with a condenser, a temperature probe, a N2 gas inlet, and a mechanical stirrer. The reaction vessel was purged with N2 for at least 15 minutes and then charged with 4-amino-5-fluoro-3-[6- (4-methyl-piperazin-l-yl)-lH-benzimidazol-2-yl]-lH-quinolin-2-one (484 g, 1.23 mol). A solution of D,L-Lactic acid (243.3 g, 1.72 mol of monomer-see the following paragraph), water (339 mL), and ethanol (1211 mL) was prepared and then charged to the reaction flask. Stirring was initiated at a medium rate, and the reaction was heated to an internal temperature of 68-72°C. The internal temperature of the reaction was maintained at 68-72°C for 15-45 minutes and then heating was discontinued. The resulting mixture was filtered through a 10-20 micron frit collecting the filtrate in a 12 L flask. The 12 L flask was equipped with an internal temperature probe, a reflux condenser, an addition funnel, a gas inlet an outlet, and an overhead stirrer. The filtrate was then stirred at a medium rate and heated to reflux (internal temperature of about 78°C). While maintaining a gentle reflux, ethanol (3,596 mL) was charged to the flask over a period of about 20 minutes. The reaction flask was then cooled to an internal temperature ranging from about 64-70°C within 15-25 minutes and this temperature was maintained for a period of about 30 minutes. The reactor was inspected for crystals. If no crystals were present, then crystals of the lactic acid salt of 4-amino-5-fluoro-3- [6-(4-methyl-piperazin- 1 -yl)- 1 H-benzimidazol -2-yl] - 1 H- quinolin-2-one (484 mg, 0.1 mole %) were added to the flask, and the reaction was stirred at 64-70°C for 30 minutes before again inspecting the flask for crystals. Once crystals were present, stirring was reduced to a low rate and the reaction was stirred at 64-70°C for an additional 90 minutes. The reaction was then cooled to about 0°C over a period of about 2 hours, and the resulting mixture was filtered through a 25-50 micron fritted filter. The reactor was washed with ethanol (484 mL) and stirred until the internal temperature was about 0°C. The cold ethanol was used to wash the filter cake, and this procedure was repeated 2 more times. The collected solid was dried to a constant weight at 50°C under vacuum in a vacuum oven yielding 510.7 g (85.7%) of the crystalline yellow lactic acid salt of 4-amino-5-fluoro-3-[6-(4-methyl-piperazin- l-yl)-lH-benzimidazol-2-yl]-lH-quinolin-2-one. A rubber dam or inert conditions were typically used during the filtration process. While the dry solid did not appear to be very hygroscopic, the wet filter cake tends to pick up water and become sticky. Precautions were taken to avoid prolonged exposure of the wet filter cake to the atmosphere.
[0155] Commercial lactic acid generally contains about 8-12% w/w water, and contains dimers and trimers in addition to the monomeric lactic acid. The mole ratio of lactic acid dimer to monomer is generally about 1.0:4.7. Commercial grade lactic acid may be used in the process described in the preceding paragraph as the monolactate salt preferentially precipitates from the reaction mixture.
Identification of Metabolites
[0156] Two metabolites of 4-amino-5-fluoro-3-[6-(4-methyl-piperazin-l-yl)- lH-benzimidazol-2-yl]-lH-quinolin-2-one (Compound 1) have been identified and characterized in pooled rat plasma from a 2 week toxicology study as described in the references incorporated herein. The two identified metabolites were the piperazine N- oxide compound (Compound 2) and the N-demethylated compound (Compound 3) shown below.
Compound 2
Figure imgf000108_0002
Compound 3
IC50S of Compounds 1-3 [0157] The kinase activity of a number of protein tyrosine kinases was measured using the procedures described in the various references incorporated herein. Some of these are shown in the following Table..
Table. IC50S of Compounds 1-3
Figure imgf000109_0002
Synthesis of 4-Amino-5-fluoro-3-[6-(4-methyl-4-oxidopiperazin-l-yl)-lH- benzimidazol-2-yl]quinolin-2(lH)-one (Compound 2) and 4-Amino-5-fluoro-3-(6- piperazin-l-yl-lH-benzimidazol-2-yl)quinolin-2(lH)-one (Compound 3) [0158] To confirm the structures of the identified metabolites of Compound 1, the metabolites were independently synthesized.
[0159] Compound 2, the N-oxide metabolite of Compound 1, was synthesized as shown in the scheme below. Compound 1 was heated in a mixture of ethanol, dimethylacetamide and hydrogen peroxide. Upon completion of the reaction, Compound 2 was isolated by filtration and washed with ethanol. If necessary, the product could be further purified by column chromatography.
Figure imgf000109_0001
[0160] Compound 3, the N-desmethyl metabolite of Compound 1, was synthesized as shown in the scheme below. 5-Chloro-2-nitroaniline was treated with piperazine to yield 4 which was subsequently protected with a butyloxycarbonyl (Boc) group to yield 5. Reduction of the nitro group followed by condensation with 3-ethoxy-3-iminopropionic acid ethyl ester gave 6. Condensation of 6 with 6- fluoroanthranilonitrile using potassium hexamethyldisilazide as the base yielded 7. Crude 7 was treated with aqueous HCl to yield the desired metabolite as a yellow/brown solid after purification.
Figure imgf000110_0001
[0161] To identify plasma biomarkers of 4-amino-5-fluoro-3-[6-(4- methylpiperazin-l-yl)-lH-benzimidazol-2-yl]quinolin-2(lH)-one treatment, the 4T1 spontaneously metastatic mouse breast tumor model was used, and circulating serum markers were analyzed by ELISA.
[0162] 4T1 breast tumor cells were grown as subcutaneous tumors in BALB/C mice, and treatment (10, 30, 60, 100, and 150 mg/kg) with 4-amino-5-fluoro-3-[6-(4- methylpiperazin-l-yl)-lH-benzimidazol-2-yl]quinolin-2(lH)-one (Compound 1) were initiated when tumors were approximately 150 mm3. Mice were dosed orally, daily for 18 days.
[0163] The serum was collected from individual animals after 18 days, and the levels of circulating cell adhesion molecules, soluble ICAM, VCAM, and E-selectin, were measured by ELISA assay. [0164] Figure 1 is a graph showing the effects of 4-amino-5-fluoro-3-[6-(4- methylpiperazin-l-yl)-lH-benzimidazol-2-yl]quinolin-2(lH)-one in the 4T1 murine breast tumor model. The growth of subcutaneous tumors was inhibited (40-80% compared to control), liver metastases were completely inhibited, and lung metastases were inhibited by 60-97% after 18 days of dosing. Various data regarding the incidence of metastases is shown in Figure 1 and included in the following Table
Figure imgf000111_0001
[0165] Wells of a Nunc Maxisorb "U" bottom microtiter plate (#449824) were coated with monoclonal capture antibody, rat anti-mouse VCAM-1 (R&D Systems #BCA12), at 5 μg/mL in phosphate buffered saline (PBS), 50 μL/well, and incubated at 37°C for 1 hour. The plates were washed 3 times with wash buffer [PBS containing 0.1% Tween 20 and 1% goat serum (Gibco BRL #16210-072)]. Wells were blocked with 150 μL/well wash buffer and incubated at 37°C for 1 hour. The blocking solution was removed from the wells and the standard (recombinant mouse VCAM-1/Fc Chimera NOS derived R&D Systems #643 -VM) and samples were diluted in wash buffer and added to the wells.
[0166] The standard was used at a range of 4000 pg/mL to 31 pg/mL. The serum samples were diluted 1/200 followed by 3-fold serial dilutions. The samples and standards were added at 50 μL/well and incubated at 37°C for 1 hour. The plates were washed three times and incubated at 37°C for 1 hour with the primary antibody (biotinylated goat anti-mouse VCAM-1, R&D Systems #BAF643) diluted 1/200 in wash buffer, 50 μL/well. The plates were washed as described above and incubated at 37°C for 1 hour with stepavidin-HRP (R&D Systems #DY998) 1/200 in PBS/1% goat serum without Tween 20.
[0167] The plates were washed three times with wash buffer and three times with PBS. They were then developed with TMB substrate (Kirkegaard & Perry labs # 50-76-00) 50 μL/well and incubated at room temperature for 10 minutes. The reaction was stopped with the addition of 50 μL/well 4N H SO4, and the plates were read at 450-550 dual wavelength on the Molecular Devices Vmax plate reader.
[0168] Wells of a Nunc Maxisorb "U" bottom microtiter plate (#449824) were coated with monoclonal capture antibody, rat anti-mouse ICAM-1 (R&D Systems #BSA2), at 5 μg/mL in phosphate buffered saline (PBS), 50 μL/well and incubated at 37°C for 1 hour. The plates were washed 3 times with wash buffer [PBS containing 0.1% Tween 20 and 1% Carnation Nonfat Dry Milk]. Wells were blocked with 150 μL/well wash buffer and incubated at 37°C for 1 hour. The blocking solution was removed from the wells and the standard (a pool of serum from mice implanted with KM12L4a or 4T1 tumors) and samples were diluted in wash buffer and added to the wells.
[0169] The standard was used at a dilution range of 1/10-1/1280. The serum samples were diluted 1/15 followed by 3-fold serial dilutions. The samples and standards were added at 50 μL/well and incubated at 37°C for 1 hour. The plates were washed three times and incubated at 37°C for 1 hour with the primary antibody (goat anti-ICAM-1, Santa Cruz Biotechnology #sc-1511) diluted 1/250 in wash buffer, 50 μL/well. The plates were washed as above and incubated at 37°C for 1 hour with 50 μL/well of the secondary antibody (swine anti-goat IgG HRPO labeled, Caltag #G50007) 1/2000 in wash buffer.
[0170] The plates were washed three times with wash buffer and three times with PBS, then developed with TMB substrate (Kirkegaard & Perry labs # 50-76-00) 50 μL/well and incubated at room temperature for 10 minutes. The reaction was stopped with the addition of 50 μL/well 4N H2SO , and the plates were read at 450- 550 dual wavelength on the Molecular Devices Vmax plate reader. [0171] Serum samples were assayed by the R&D Systems Quantikine M,
Mouse sE-Selectin Immunoassay kit #MESOO according to the manufacturer's protocol.
In Vivo KM12L4a Human Colon Xenografts
[0172] Female Nu/nu mice (6-8 weeks old, 18-22 grams) were obtained from
Charles River Laboratories (Wilmington, MA). Tumor cells (2 x 106KM12L4a) were implanted subcutaneous into the flank of mice and allowed to grow to the desired size before treatment was initiated. Tumor bearing mice were administered with 100 mg/kg of 4-amino-5-fluoro-3-[6-(4-methylpiperazin-l-yl)-lH-benzimidazol-2- yl]quinolin-2(lH)-one for 7 days, and individual mice were euthanized. The tumors were resected and flash frozen in liquid nitrogen.
Zvmography for MMP-2 and MMP-9 Activity
[0173] Resected tumors were lysed in RIPA buffer (1 % Nonidet P-40, 0.5% sodium deoxycholate, 0.1% Sodium dodecylsulphate in IX phosphate buffered saline, pH 7.2) containing protease inhibitors (Roche Molecular Biochemicals) and phosphatase inhibitors (Sigma). 50 μg of total proteins were analyzed by gelatin zymography on 12% SDS polyacrylamide with gelatin substrate. After electrophoresis, gels were washed twice for 15 minutes in 2.5% Triton X-100, incubated overnight at 37°C in 50 mM Tris-HCl and 10 mM CaCl2, pH 7.6, and stained with 0.5% Comassie Blue and destained with 50% methanol.
ELISA
[0174] VEGF-A protein levels in KM12L4a tumor lysates were quantified using a commercially available ELISA kit (R and D Systems, Minneapolis, MN) according to the manufacturer's procedures.
[0175] Analysis of KM12L4a human colon tumors, removed after in vivo administration of 4-amino-5-fluoro-3 - [6-(4-methy lpiperazin- 1 -yl)- 1 H-benzimidazol- 2-yl]quinolin-2(lH)-one, showed reduced VEGF production and decreased MMP-9 activity. These changes were accompanied by decreased tumor cell proliferation ill (Ki67), induced apoptosis (increased PARP cleavage and caspase-3) and reduced vascular density (CD31) as seen by antibody immunohistochemistry staining.
[0176] The preparation of numerous quinolinone benzimidazole compounds useful in inhibiting angiogenesis and vascular endothelial growth factor receptor tyrosine kinases and in inhibiting other tyrosine and serine/threonine kinases including 4-amino-5-fluoro-3 - [5-(4-methylpiperazin- 1 -yl)- 1 H-benzimidazol-2- yl]quinolin-2(lH)-one or a tautomer thereof is disclosed in the following documents which are each hereby incorporated by reference in their entireties and for all purposes as if fully set forth herein: U.S. Patent No. 6,605,617; U.S. Patent No. 6,756,383; U.S. Patent Application No. 10/116,117 filed (published on February 6, 2003, as US 2003/0028018 Al); U.S. Patent Application No. 10/644,055 (published on May 13, 2004, U.S. Patent Application No. 2004/0092535); U.S. Patent Application No. 10/983,174; U.S. Patent Application No. 10/706,328 (published on November 4, 2004, as 2004/0220196); U.S. Patent Application No. 10/982,757; and U.S. Patent Application No. 10/982,543..
Western Blot Analysis
[0177] HUVECs were cultured in EGM (Endothelial Cell Growth Media) with or without 100 nM 4-amino-5-fluoro-3-[6-(4-methylpiperazin-l-yl)-lH- benzimidazol-2-yl]quinolin-2(lH)-one (Compound 1), and cell lysates were collected at 0, 16, and 24 hours post-treatment. Equal amounts of proteins were loaded in 4- 20% SDS-PAGE, and the gels were probed with antibodies against ICAM, VCAM, α5 integrin,, and αv integrin. The equal loading and efficiency was evaluated by probing with anti β-actin antibody. The expression of ICAM, VCAM, and α5 integrin was decreased with 4-amino-5-fluoro-3-[6-(4-methylpiperazin-l-yl)-lH- benzimidazol-2-yl]quinolin-2(lH)-one treatment in HUVECs in vitro.
[0178] It should be understood that the organic compounds according to the invention may exhibit the phenomenon of tautomerism. As the chemical stnictures within this specification can only represent one of the possible tautomeric forms at a time, it should be understood that the invention encompasses any tautomeric form of the drawn structure. For example, the compound of Structure IIIB is shown below with one tautomer, Tautomer IIIBa:
Figure imgf000115_0001
IIIB
Figure imgf000115_0002
Tautomer IIIBa
[0179] Other tautomers of the compound of Structure IIIB, Tautomer IIIBb and Tautomer IIIBc, are shown below:
Figure imgf000116_0001
Tautomer IIIBb
Figure imgf000116_0002
Tautomer IIIBc
[0180] All documents or references cited herein are hereby incorporated by reference in their entireties and for all purposes as if fully set forth herein.
[0181] It is understood that the invention is not limited to the embodiments set forth herein for illustration, but embraces all such forms thereof as come within the scope of the invention.

Claims

What is claimed is: 1. A method of modulating an inflammatory response and/or reducing cellular adhesion in a subject, comprising: administering to the subject a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof, wherein the inflammatory response is modulated in the subject and/or cellular adhesion is reduced in the subject after administration, and Stnicture I has the following formula:
Figure imgf000117_0001
I wherein, R1, R2, R3, and R4 may be the same or different and are independently selected from the group consisting of H, Cl, Br, F, I, -CN, -NO2, -OH, -OR15 groups, -NR16R17 groups, substituted and unsubstituted amidinyl groups, substituted and unsubstituted guanidinyl groups, substituted and unsubstituted primary, secondary, and tertiary alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted alkenyl groups, substituted and unsubstituted alkynyl groups, substituted and unsubstituted heterocyclyl groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted heterocyclylalkyl groups, and -C(=O)R18 groups; R5, R6, R7, and R8 may be the same or different and are independently selected from the group consisting of H, Cl, Br, F, I, -NO2, -OH, -OR19 groups, -NR20R21 groups, -SH, -SR22 groups, -S(=O)R23 groups, -S(=O)2R24 groups, -CN, substituted and unsubstituted amidinyl groups, substituted and unsubstituted guanidinyl groups, substituted and unsubstituted primary, secondary, and tertiary alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted alkenyl groups, substituted and unsubstituted alkynyl groups, substituted and unsubstituted heterocyclyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryι)aminoalkyl groups, substituted and unsubstituted heterocyclylalkyl groups, -C(=O)R25 groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted heterocyclylaminoalkyl groups, substituted and unsubstituted hydroxyalkyl groups, substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted aryloxyalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups; R12 is selected from the group consisting of H, substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, and substituted and unsubstituted heterocyclyl groups; R13 is selected from the group consisting of H, substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted heterocyclyl groups, -OH, alkoxy groups, aryloxy groups, -NH2, substituted and unsubstituted heterocyclylalkyl groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted alkylamino groups, substituted and unsubstituted arylamino groups, substituted and unsubstituted dialkylamino groups, substituted and unsubstituted diarylamino groups, substituted and unsubstituted (alkyl)(aryl)amino groups, -C(=O)H, -C(=O)-alkyl groups, -C(=O)-aryl groups, -C(=O)O-alkyl groups, -C(=O)O-aryl groups, -C(=O)NH2, -C(=O)NH(alkyl) groups, -C(=O)NH(aryl) groups, -C(=O)N(alkyl)2 groups, -C(=O)N(aryl)2 groups, -C(=O)N(alkyl)(aryl) groups, -C(=O)-heterocyclyl groups, -C(=:O)-O-heterocyclyl groups, -C(=O)NH(heterocyclyl) groups, -C(=O)-N(heterocyclyι)2 groups, -C(=O)-N(alkyι)(heterocyclyι) groups, -C(=O N(aryl)(heterocyclyl) groups, substituted and unsubstituted heterocyclylaminoalkyl groups, substituted and unsubstituted hydroxyalkyl groups, substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted aryloxyalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups; R15 and R19 may be the same or different and are independently selected from the group consisting of substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted heterocyclyl groups, substituted and unsubstituted heterocyclylalkyl groups, -C(=O)H, -C(=O)-alkyl groups, -C(=O)-aryl groups, -C(=O)NH2, -C(=O)NH(alkyl) groups, -C(=O)NH(aryl) groups, -C(=O)N(alkyl)2 groups, -C(=O)N(aryl)2 groups, -C(=O)N(alkyl)(aryl) groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted heterocyclylaminoalkyl, substituted and unsubstituted diheterocyclylammoalkyl, substituted and unsubstituted (heterocyclyl)(alkyl)aminoalkyl, substituted and unsubstituted (heterocyclyl)(aryl)aminoalkyl, substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted hydroxyalkyl groups, substituted and unsubstituted aryloxyalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups; R16 and R20 may be the same or different and are independently selected from the group consisting of H, substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, and substituted and unsubstituted heterocyclyl groups; R17 and R21 may be the same or different and are independently selected from the group consisting of H, substituted and unsubstituted alkyl groups, substituted and 82 unsubstituted aryl groups, substituted and unsubstituted heterocyclyl groups, 83 -C(=O)H, -C(=O)-alkyl groups, -C(=O)-aryl groups,-C(=O)NH2, -C(=O)NH(alkyl) 84 groups, -C(=O)NH(aryl) groups, -C(=O)N(alkyl)2 groups, -C(=O)N(aryl)2 groups, 85 -C(=O)N(alkyl)(aryl) groups, -C(=O)O-alkyl groups, -C(=O)O-aryl groups, 86 substituted and unsubstituted heterocyclylalkyl groups, substituted and unsubstituted 87 aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted 88 and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted 89 arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, 90 substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, -C(=O)-heterocyclyl 91 groups, -C(=O)-O-heterocyclyl groups, -C(=O)NH(heterocyclyl) groups, 92 -C(=O)-N(heterocyclyl) groups, -C(=O)-N(alkyl)(heterocyclyl) groups, 93 -C(=O)-N(aryl)(heterocyclyl) groups, substituted and unsubstituted 94 heterocyclylaminoalkyl groups, substituted and unsubstituted hydroxyalkyl groups, 95 substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted 96 aryloxyalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups; 97 R , R , R , and R may be the same or different and are independently 98 selected from the group consisting of H, -NH2, -NH(alkyl) groups, -NH(aryl) groups, 99 -N(alkyl)2 groups, -N(aryl)2 groups, -N(alkyl)(aryl) groups, -NH(heterocyclyl)
100 groups, -N(heterocyclyl)(alkyl) groups, -N(heterocyclyl)(aryl) groups,
101 -N(heterocyclyl)2 groups, substituted and unsubstituted alkyl groups, substituted and
102 unsubstituted aryl groups, -OH, substituted and unsubstituted alkoxy groups,
103 substituted and unsubstituted aryloxy groups, substituted and unsubstituted
104 heterocyclyl groups, -NHOH, -N(alkyl)OH groups, -N(aryl)OH groups,
105 -N(alkyl)O-alkyl groups, -N(aryl)O-alkyl groups, -N(alkyl)O-aryl groups, and
106 -N(aryl)O-aryl groups; and
107 R22 is selected from the group consisting of substituted and unsubstituted alkyl
108 groups, substituted and unsubstituted aryl groups, and substituted and unsubstituted
109 heterocyclyl groups.
1 2. The method of claim 1 , wherein the levels of circulating 2 adhesion molecules are reduced in the subject after administration.
3. The method of claim 1 , wherein the amount of at least one of inducible cell adhesion molecule, vascular cell adhesion molecule, or endothelial leukocyte adhesion molecule is reduced in the subject after administration.
4. The method of claim 1 , further comprising: measuring the amount of at least one of inducible cell adhesion molecule, vascular cell adhesion molecule, or endothelial leukocyte adhesion molecule in the subject after administration of the compound, the tautomer, the pharmaceutically acceptable salt of the compound, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof.
5. The method of claim 1 , wherein the amount of a matrix metalloprotease is reduced in the subject after administration.
6. The method of claim 5, wherein the matrix metalloprotease is matrix metalloprotease-2 or matrix metalloprotease-9.
7. The method of claim 1 , wherein the compound of Structure I is a compound of Structure II, and Stracture II has the following formula:
Figure imgf000121_0001
II wherein,
A is a group having one of the following Structures:
Figure imgf000122_0001
wherein,
Ra is selected from H or straight or branched chain alkyl groups having from 1 to 6 carbon atoms.
8. The method of claim 7, wherein Ra is a methyl group, and the compound of Structure II has the Stracture IIA having the following formula:
Figure imgf000122_0002
IIA
9. The method of claim 8, wherein the pharmaceutically acceptable salt of the compound of Stracture IIA, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof is administered to the subject, and the salt is a lactate salt. 10. The method of claim 7, wherein Ra is a hydrogen, and the compound of Structure II has the Stracture IIB having the following formula:
Figure imgf000123_0001
ΠB 11. The method of claim 7, wherein Ra is a methyl group, and the compound of Stracture II has the Stracture IIC having the following formula:
Figure imgf000123_0002
IIC
12.- The method of claim 1, wherein the pharmaceutically acceptable salt of the compound of Stracture I, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof is administered to the subject, and the salt is a lactate salt.
13. The method of claim 1 , wherein the subject is a cancer patient.
14. The method of claim 1 , wherein R and R are both H.
15. The method of claim 1, wherein at least one of
Figure imgf000123_0003
is a substituted or unsubstituted heterocyclyl group.
16. The method of claim 1, wherein at least one of R or R is a substituted or unsubstituted piperazine group.
17. The method of claim 1 , wherein R1 is F and R2, R3, R4, R5, and R8 are each H and one of R6 or R7 is H.
18. A method of identifying a subj ect in need of a compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof, comprising measuring the amount of at least one cell adhesion molecule in the subject before, during, or after administration of the compound of Stracture I, the tautomer of the compound, the pharmaceutically acceptable salt of the compound, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof to the subject, wherein the cell adhesion molecule is selected from inducible cell adhesion molecule, vascular cell adhesion molecule, or endothelial leukocyte adhesion molecule, and Stracture I has the following formula:
Figure imgf000124_0001
I wherein, R1, R2, R3, and R4 may be the same or different and are independently selected from the group consisting of H, Cl, Br, F, I, -CN, -NO2, -OH, -OR15 groups, -NR16R17 groups, substituted and unsubstituted amidinyl groups, substituted and unsubstituted guanidinyl groups, substituted and unsubstituted primary, secondary, and tertiary alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted alkenyl groups, substituted and unsubstituted alkynyl groups, substituted and unsubstituted heterocyclyl groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted heterocyclylalkyl groups, and -C(=O)R groups; R5, R6, R7, and R8 may be the same or different and are independently selected from the group consisting of H, Cl, Br, F, I, -NO2, -OH, -OR19 groups, -NR20R21 groups, -SH, -SR22 groups, -S(=O)R23 groups, -S(=O)2R24 groups, -CN, substituted and unsubstituted amidinyl groups, substituted and unsubstituted guanidinyl groups, substituted and unsubstituted primary, secondary, and tertiary alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted alkenyl groups, substituted and unsubstituted alkynyl groups, substituted and unsubstituted heterocyclyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and 9 _r unsubstituted heterocyclylalkyl groups, -C(=O)R groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted heterocyclylaminoalkyl groups, substituted and unsubstituted hydroxyalkyl groups, substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted aryloxyalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups; R12 is selected from the group consisting of H, substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, and substituted and unsubstituted heterocyclyl groups; R13 is selected from the group consisting of H, substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted heterocyclyl groups, -OH, alkoxy groups, aryloxy groups, -NH2, substituted and unsubstituted heterocyclylalkyl groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted alkylamino groups, substituted and unsubstituted arylamino groups, substituted and unsubstituted dialkylamino groups, substituted and unsubstituted diarylamino groups, substituted and unsubstituted (aIkyι)(aryl)amino groups, -C(=O)H, -C(=0)-alkyl groups, -C(=O)-aryl groups, -C(=O)O-alkyl groups, -C(=O)O-aryl groups, -C(=O)NH2, -C(=0)NH(alkyl) groups, -C(=O)NH(aryl) groups, -C(=0)N(alkyl)2 groups, -C(=O)N(aryl)2 groups, -C(=O)N(alkyl)(aryl) groups, -C(=O)-heterocyclyl groups, -C(=O)-O-heterocyclyl groups, -C(=O)NH(heterocyclyl) groups, -C(=O)-N(heterocyclyl)2 groups, -C(=O)-N(alkyl)(heterocyclyl) groups, -C(=O)- N(aryl)(heterocyclyl) groups, substituted and unsubstituted heterocyclylaminoalkyl groups, substituted and unsubstituted hydroxyalkyl groups, substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted aryloxyalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups; R15 and R19 may be the same or different and are independently selected from the group consisting of substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted heterocyclyl groups, substituted and unsubstituted heterocyclylalkyl groups, -C(=O)H, ~C(=O)~alkyl groups, -C(=O)-aryl groups, -C(=O)NH2, -C(=O)NH(alkyl) groups, -C(=O)NH(aryl) groups, -C(=O)N(alkyl)2 groups, -C(=O)N(aryl)2 groups, -C(=O)N(alkyl)(aryl) groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted heterocyclylaminoalkyl, substituted and unsubstituted diheterocyclylaminoalkyl, substituted and unsubstituted (heterocyclyl)(alkyl)aminoalkyl, substituted and unsubstituted (heterocyclyt)(aryl)aminoalkyl, substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted hydroxyalkyl groups, substituted and unsubstituted aryloxyalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups; R16 and R20 may be the same or different and are independently selected from the group consisting of H, substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, and substituted and unsubstituted heterocyclyl groups; 83 R17 and R21 may be the same or different and are independently selected from 84 the group consisting of H, substituted and unsubstituted alkyl groups, substituted and 85 unsubstituted aryl groups, substituted and unsubstituted heterocyclyl groups, 86 -C(=O)H, -C(=O)-alkyl groups, -C(=O)-aryl groups,-C(=O)NH2, -C(=O)NH(alkyl) 87 groups, -C(=O)NH(aryl) groups, -C(=O)N(alkyl)2 groups, -C(=O)N(aryl)2 groups, 88 -C(=O)N(alkyl)(aryl) groups, -C(=O)O-alkyl groups, -C(=O)O-aryl groups, 89 substituted and unsubstituted heterocyclylalkyl groups, substituted and unsubstituted 90 aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted 91 and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted 92 arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, 93 substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, -C(=O)-heterocyclyl 94 groups, -C(=O)-O-heterocyclyl groups, -C(=O)NH(heterocyclyl) groups, 95 -C(=O)-N(heterocyclyl)2 groups, -C(=O)-N(alkyl)(heterocyclyl) groups, 96 -C(=O)-N(aryl)(heterocyclyl) groups, substituted and unsubstituted 97 heterocyclylaminoalkyl groups, substituted and unsubstituted hydroxyalkyl groups, 98 substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted 99 aryloxyalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups;
100 R18, R23, R24, and R25 may be the same or different and are independently
101 selected from the group consisting of H, -NH2, -NH(alkyl) groups, -NH(aryl) groups,
102 -N(alkyl)2 groups, -N(aryl)2 groups, -N(alkyl)(aryl) groups, -NH(heterocyclyl)
103 groups, -N(heterocyclyl)(alkyl) groups, -N(heterocyclyl)(aryl) groups,
104 -N(heterocyclyl)2 groups, substituted and unsubstituted alkyl groups, substituted and
105 unsubstituted aryl groups, -OH, substituted and unsubstituted alkoxy groups,
106 substituted and unsubstituted aryloxy groups, substituted and unsubstituted
107 heterocyclyl groups, -NHOH, -N(alkyl)OH groups, -N(aryl)OH groups,
108 -N(alkyl)O-alkyl groups, -N(aryl)O-alkyl groups, -N(alkyl)O-aryl groups, and
109 -N(aryl)O-aryl groups; and
110 R22 is selected from the group consisting of substituted and unsubstituted alkyl
111 groups, substituted and unsubstituted aryl groups, and substituted and unsubstituted
112 heterocyclyl groups .
19. The method of claim 18, wherein the compound of Stracture I is a compound of Structure II, and Structure II has the following formula:
Figure imgf000128_0001
II wherein,
A is a group having one of the following Structures:
Figure imgf000128_0002
wherein,
Ra is selected from H or straight or branched chain alkyl groups having from 1 to 6 carbon atoms.
20. The method of claim 19, wherein Ra is a methyl group, and the compound of Stracture II has the Structure IIA having the following formula:
Figure imgf000128_0003
IIA
21. The method of claim 20, wherein the pharmaceutically acceptable salt of the compound of Stracture IIA, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof is administered to the subject, and the salt is a lactate salt.
22. The method of claim 19, wherein Ra is a hydrogen, and the compound of Structure II has the Stracture IIB having the following formula:
Figure imgf000129_0001
IIB
23. The method of claim 19, wherein Ra is a methyl group, and the compound of Stracture II has the Stracture IIC having the following formula:
Figure imgf000129_0002
IIC
24. The method of claim 18, wherein the pharmaceutically acceptable salt of the compound of Structure I, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof is administered to the subject, and the salt is a lactate salt.
25. The method of claim 18, wherein the patient is a cancer patient.
26. The method of claim 18, wherein R12 and R13 are both H.
27. The method of claim 18, wherein at least one of R6 or R7 is a substituted or unsubstituted heterocyclyl group.
28. The method of claim 18, wherein at least one of R6 or R7 is a substituted or unsubstituted piperazine group.
29. The method of claim 18, wherein R1 is F and R2, R3, R4, R5, and R8 are each H and one of R6 or R7 is H.
30. The method of claim 18, further comprising withdrawing a sample of blood from the subject and then measuring the amount of the at least one cell adhesion molecule in at least a portion of the sample.
31. A method of monitoring the progression of a disease or treatment in a subject, comprising: measuring the amount of at least one cell adhesion molecule in the subject after administration ofa compound of Structure I, a tautomer of the compound, a pharmaceutically acceptable salt of the compound, a pharmaceutically acceptable salt of the tautomer, or a mixture thereof to the subject, wherein the cell adhesion molecule is selected from inducible cell adhesion molecule, vascular cell adhesion molecule, or endothelial leukocyte adhesion molecule, and Structure I has the following formula:
Figure imgf000130_0001
wherein, R1, R2, R3, and R4 may be the same or different and are independently selected from the group consisting of H, Cl, Br, F, I, -CN, -NO2, -OH, -OR15 groups, -NR16R17 groups, substituted and unsubstituted amidinyl groups, substituted and unsubstituted guanidinyl groups, substituted and unsubstituted primary, secondary, and tertiary alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted alkenyl groups, substituted and unsubstituted alkynyl groups, substituted and unsubstituted heterocyclyl groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyiχaryl)aminoalkyl groups, substituted and unsubstituted heterocyclylalkyl groups, and -C(=O)R18 groups; R5, R6, R7, and R8 may be the same or different and are independently selected from the group consisting of H, Cl, Br, F, I, -NO2, -OH, -OR19 groups, -NR20R21 groups, -SH, -SR22 groups, -S(=O)R23 groups, -S(=O)2R24 groups, -CN, substituted and unsubstituted amidinyl groups, substituted and unsubstituted guanidinyl groups, substituted and unsubstituted primary, secondary, and tertiary alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted alkenyl groups, substituted and unsubstituted alkynyl groups, substituted and unsubstituted heterocyclyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted heterocyclylalkyl groups, -C(=O)R25 groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted heterocyclylaminoalkyl groups, substituted and unsubstituted hydroxyalkyl groups, substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted aryloxyalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups; R12 is selected from the group consisting of H, substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, and substituted and unsubstituted heterocyclyl groups; R13 is selected from the group consisting of H, substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted heterocyclyl groups, -OH, alkoxy groups, aryloxy groups, -NH2, substituted and unsubstituted heterocyclylalkyl groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, substituted and unsubstituted alkylamino groups, substituted and unsubstituted arylamino groups, substituted and unsubstituted dialkylamino groups, substituted and unsubstituted diarylamino groups, substituted and unsubstituted (alkyl)(aryl)amino groups, -C(=O)H, -C(=O)-alkyl groups, -C(=O)-aryl groups, -C(=O)O-alkyl groups, -C(=O)O-aryl groups, -C(=0)NH2, -C(=0)NH(alkyl) groups, -C(=O)NH(aryl) groups, -C(=O)N(alkyl)2 groups, -C(=O)N(aryl)2 groups, -C(=O)N(alkyl)(aryl) groups, -C(=O)-heterocyclyl groups, -C(=O)-O-heterocyclyl groups, -C(=O)NH(heterocyclyl) groups, -C(=O)-N(heterocyclyl)2 groups, -C(=O)-N(alkyl)(heterocyclyl) groups, -C(=O)- N(aryl)(heterocyclyι) groups, substituted and unsubstituted heterocyclylaminoalkyl groups, substituted and unsubstituted hydroxyalkyl groups, substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted aryloxyalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups; R15 and R19 may be the same or different and are independently selected from the group consisting of substituted and unsubstituted alkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted heterocyclyl groups, substituted and unsubstituted heterocyclylalkyl groups, -C(=O)H, -C(=O)-alkyl groups, -C(=O)-aryl groups, -C(=O)NH2, -C(=O)NH(alkyl) groups, -C(=O)NH(aryl) groups, -C(=O)N(alkyl)2 groups, -C(=O)N(aryl)2 groups, -C(=O)N(alkyl)(aryl) groups, substituted and unsubstituted aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, substituted and unsubstituted (alkyl)(aryι)aminoalkyl groups, substituted and unsubstituted heterocyclylaminoalkyl, substituted and unsubstituted diheterocyclylaminoalkyl, substituted and unsubstituted 74 (heterocyclyl)(alkyl)aminoalkyl, substituted and unsubstituted 75 (heterocyclyl)(aryl)aminoalkyl, substituted and unsubstituted alkoxyalkyl groups, 76 substituted and unsubstituted hydroxyalkyl groups, substituted and unsubstituted 77 aryloxyalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups; 78 R16 and R20 may be the same or different and are independently selected from 79 the group consisting of H, substituted and unsubstituted alkyl groups, substituted and 80 unsubstituted aryl groups, and substituted and unsubstituted heterocyclyl groups; 81 R17 and R21 may be the same or different and are independently selected from 82 the group consisting of H, substituted and unsubstituted alkyl groups, substituted and 83 unsubstituted aryl groups, substituted and unsubstituted heterocyclyl groups, 84 -C(=O)H, -C(=O)-alkyl groups, -C(=O)-aryl groups,-C(=O)NH2, -C(=O)NH(alkyl) 85 groups, -C(=O)NH(aryl) groups, -C(=O)N(alkyl)2 groups, -C(=O)N(aryl)2 groups, 86 -C(=O)N(alkyl)(aryl) groups, -C(=O)O-alkyl groups, -C(=O)O-aryl groups, 87 substituted and unsubstituted heterocyclylalkyl groups, substituted and unsubstituted 88 aminoalkyl groups, substituted and unsubstituted alkylaminoalkyl groups, substituted 89 and unsubstituted dialkylaminoalkyl groups, substituted and unsubstituted 90 arylaminoalkyl groups, substituted and unsubstituted diarylaminoalkyl groups, 91 substituted and unsubstituted (alkyl)(aryl)aminoalkyl groups, -C(=O)-heterocyclyl 92 groups, -C(=O)-O-heterocyclyl groups, -C(=O)NH(heterocyclyl) groups, 93 -C(=O)-N(heterocyclyl)2 groups, -C(=O)-N(alkyl)(heterocyclyl) groups, 94 -C(=O)-N(aryl)(heterocyclyl) groups, substituted and unsubstituted 95 heterocyclylaminoalkyl groups, substituted and unsubstituted hydroxyalkyl groups, 96 substituted and unsubstituted alkoxyalkyl groups, substituted and unsubstituted 97 aryloxyalkyl groups, and substituted and unsubstituted heterocyclyloxyalkyl groups; 98 R18, R23, R24, and R25 may be the same or different and are independently 99 selected from the group consisting of H, -NH2, -NH(alkyl) groups, -NH(aryl) groups,
100 -N(alkyl)2 groups, -N(aryl)2 groups, -N(alkyl)(aryl) groups, -NHCheterocyclyl)
101 groups, -N(heterocyclyι)(alkyι) groups, -N(heterocyclyl)(aryl) groups,
102 -N(heterocyclyl)2 groups, substituted and unsubstituted alkyl groups, substituted and
103 unsubstituted aryl groups, -OH, substituted and unsubstituted alkoxy groups,
104 substituted and unsubstituted aryloxy groups, substituted and unsubstituted 105 heterocyclyl groups, -NHOH, -N(alkyl)OH groups, -N(aryl)OH groups, 106 -N(alkyl)O-alkyl groups, -N(aryl)O-alkyl groups, -N(alkyl)O-aryl groups, and 107 -N(aryl)O-aryl groups; and
108 R22 is selected from the group consisting of substituted and unsubstituted alkyl 109 groups, substituted and unsubstituted aryl groups, and substituted and unsubstituted 110 heterocyclyl groups.
1 32. The method of claim 31 , wherein the compound of Stracture I 2 is a compound of Structure II, and Stracture II has the following formula:
Figure imgf000134_0001
II wherein,
5 A is a group having one of the following Structures:
Figure imgf000134_0002
7 wherein,
8 Ra is selected from H or straight or branched chain alkyl groups having from 1 to 6 9 carbon atoms.
10 33. The method of claim 32, wherein Ra is a methyl group, and the 11 compound of Structure II has the Structure IIA having the following formula:
Figure imgf000135_0001
IIA
34. The method of claim 33, wherein the pharmaceutically acceptable salt of the compound of Stracture IIA, the pharmaceutically acceptable salt of the tautomer, or the mixture thereof is administered to the subject, and the salt is a lactate salt.
35. The method of claim 32, wherein Ra is a hydrogen, and the compound of Stracture II has the Stracture IIB having the following formula:
Figure imgf000135_0002
IIB
36. The method of claim 32, wherein Ra is a methyl group, and the compound of Structure II has the Structure IIC having the following formula:
Figure imgf000136_0001
IIC
37. The method of claim 31 , wherein the patient is a cancer patient.
38. The method of claim 31 , wherein R 12 and j r R, 13 are both H.
39. The method of claim 31 , further comprising withdrawing a sample of blood from the subject and then measuring the amount of the at least one cell adhesion molecule in at least a portion of the sample.
PCT/US2005/005316 2004-02-20 2005-02-18 Modulation of inflammatory and metastatic processes WO2005082340A2 (en)

Priority Applications (8)

Application Number Priority Date Filing Date Title
RU2006133536/15A RU2377988C2 (en) 2004-02-20 2005-02-18 Modulation of inflammatory and metastatic processes
CA2556872A CA2556872C (en) 2004-02-20 2005-02-18 Modulation of inflammatory and metastatic processes
BRPI0507891-1A BRPI0507891A (en) 2004-02-20 2005-02-18 modulation of inflammatory and metastatic processes
JP2006554253A JP5019884B2 (en) 2004-02-20 2005-02-18 Regulation of inflammatory and metastatic processes
AU2005216904A AU2005216904B2 (en) 2004-02-20 2005-02-18 Modulation of inflammatory and metastatic processes
CN2005800095231A CN1960731B (en) 2004-02-20 2005-02-18 Method for regulating inflammatory and metastatic processes
EP05723338A EP1718306A2 (en) 2004-02-20 2005-02-18 Modulation of inflammatory and metastatic processes
IL177574A IL177574A0 (en) 2004-02-20 2006-08-17 Modulation of inflammatory and metastatic processes

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
US54639504P 2004-02-20 2004-02-20
US60/546,395 2004-02-20
US54710304P 2004-02-23 2004-02-23
US60/547,103 2004-02-23
US55477104P 2004-03-19 2004-03-19
US60/554,771 2004-03-19

Publications (2)

Publication Number Publication Date
WO2005082340A2 true WO2005082340A2 (en) 2005-09-09
WO2005082340A3 WO2005082340A3 (en) 2006-05-04

Family

ID=34916329

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2005/005316 WO2005082340A2 (en) 2004-02-20 2005-02-18 Modulation of inflammatory and metastatic processes

Country Status (10)

Country Link
US (1) US7875624B2 (en)
EP (1) EP1718306A2 (en)
JP (1) JP5019884B2 (en)
CN (1) CN1960731B (en)
AU (1) AU2005216904B2 (en)
BR (1) BRPI0507891A (en)
CA (1) CA2556872C (en)
IL (1) IL177574A0 (en)
RU (1) RU2377988C2 (en)
WO (1) WO2005082340A2 (en)

Cited By (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006077428A1 (en) * 2005-01-21 2006-07-27 Astex Therapeutics Limited Pharmaceutical compounds
WO2006127926A2 (en) * 2005-05-23 2006-11-30 Novartis Ag Crystalline and other forms of 4-amino-5-fluoro-3-[6-(4-methylpiperazin-1-yl)-1h-benzimidazol-2-yl]-1h-quinolin-2-one lactic acid salts
WO2007077961A2 (en) * 2005-12-28 2007-07-12 Takeda Pharmaceutical Company Limited Fused heterocyclic compounds and their use as mineralocorticoid receptor ligands
EP1845990A2 (en) * 2005-01-27 2007-10-24 Novartis Vaccines and Diagnostics, Inc. Treatment of metastasized tumors
JP2011515370A (en) * 2008-03-19 2011-05-19 ノバルティス アーゲー Crystal form of 4-amino-5-fluoro-3- [5- (4-methylpiperazin-1-yl) -1H-benzimidazol-2-yl] quinolin-2 (1H) -one lactate and two solvents Japanese style
WO2011128403A1 (en) * 2010-04-16 2011-10-20 Novartis Ag Organic compound for use in the treatment of liver cancer
US8048885B2 (en) 2005-12-16 2011-11-01 Novartis Ag Organic compounds
US8080563B2 (en) 2006-07-07 2011-12-20 Kalypsys Bicyclic heteroaryl inhibitors of PDE4
US8138205B2 (en) 2006-07-07 2012-03-20 Kalypsys, Inc. Heteroarylalkoxy-substituted quinolone inhibitors of PDE4
US8173667B2 (en) 2005-10-21 2012-05-08 Novartis Ag 1-aza-bicycloalkyl derivatives
US8236803B2 (en) 2002-09-04 2012-08-07 Novartis Ag Aza-bicycloalkyl ethers and their use as alpha7-nAChR agonists
US8293767B2 (en) 2005-01-21 2012-10-23 Astex Therapeutics Limited 4-(2,6-dichloro-benzoylamino)-1H-pyrazole-3-carboxylic acid piperidin-4-ylamide acid addition salts as kinase inhibitors
US8404718B2 (en) 2005-01-21 2013-03-26 Astex Therapeutics Limited Combinations of pyrazole kinase inhibitors
US8492374B2 (en) 2009-04-29 2013-07-23 Industrial Technology Research Institute Azaazulene compounds
US8609662B2 (en) 2004-07-14 2013-12-17 Novartis Ag 3-(heteroaryl-oxy)-2-alkyl-1-aza-bicycloalkyl derivatives as alpha. 7-nachr ligands for the treatment of CNS diseases
US8759346B2 (en) 2005-12-16 2014-06-24 Novartis Ag Organic compounds
US8779147B2 (en) 2003-07-22 2014-07-15 Astex Therapeutics, Ltd. 3,4-disubstituted 1H-pyrazole compounds and their use as cyclin dependent kinase and glycogen synthase kinase-3 modulators
US8933090B2 (en) 2004-06-18 2015-01-13 Novartis Ag 1-aza-bicyclo[3.3.1]nonanes
WO2019001419A1 (en) * 2017-06-27 2019-01-03 Janssen Pharmaceutica Nv New quinolinone compounds
AU2018246382B2 (en) * 2017-03-31 2022-06-16 Vivoryon Therapeutics N.V. Novel inhibitors

Families Citing this family (22)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101167573B1 (en) * 2003-11-07 2012-07-30 노바티스 백신즈 앤드 다이아그노스틱스 인코포레이티드 Pharmaceutically acceptable salts of quinolinone compounds having improved pharmaceutical properties
ES2440799T3 (en) * 2005-05-13 2014-01-30 Novartis Ag Methods to treat drug resistant cancer
JP5545925B2 (en) * 2005-05-17 2014-07-09 ノバルティス アーゲー Method for synthesizing heterocyclic compounds
PT1957074E (en) * 2005-11-29 2014-06-25 Novartis Ag Formulations of quinolinones
WO2009101018A2 (en) * 2008-02-15 2009-08-20 F. Hoffmann-La Roche Ag 3-alkyl-piperazine derivatives and uses thereof
AU2009266833B2 (en) * 2008-07-03 2013-01-31 Novartis Ag Melt granulation process
RU2012120784A (en) * 2009-11-12 2013-12-20 Селвита С.А. COMPOUND, METHOD FOR PRODUCING IT, PHARMACEUTICAL COMPOSITION, APPLICATION OF COMPOUND, METHOD FOR MODULATION OR REGULATION OF SERINE / THREONINE KINASES AND MEANS FOR MODULATING SERINE / TREONINES
CN104968200B (en) 2013-02-01 2018-03-06 维尔斯达医疗公司 Amine compounds having anti-inflammatory, antifungal, antiparasitic and anticancer activity
EP2956138B1 (en) 2013-02-15 2022-06-22 Kala Pharmaceuticals, Inc. Therapeutic compounds and uses thereof
US9688688B2 (en) 2013-02-20 2017-06-27 Kala Pharmaceuticals, Inc. Crystalline forms of 4-((4-((4-fluoro-2-methyl-1H-indol-5-yl)oxy)-6-methoxyquinazolin-7-yl)oxy)-1-(2-oxa-7-azaspiro[3.5]nonan-7-yl)butan-1-one and uses thereof
ES2831625T3 (en) 2013-02-20 2021-06-09 Kala Pharmaceuticals Inc Therapeutic compounds and their uses
KR102365952B1 (en) 2013-10-14 2022-02-22 에자이 알앤드디 매니지먼트 가부시키가이샤 Selectively substituted quinoline compounds
KR102103256B1 (en) 2013-10-14 2020-04-23 에자이 알앤드디 매니지먼트 가부시키가이샤 Selectively substituted quinoline compounds
US9890173B2 (en) 2013-11-01 2018-02-13 Kala Pharmaceuticals, Inc. Crystalline forms of therapeutic compounds and uses thereof
MX355330B (en) 2013-11-01 2018-04-16 Kala Pharmaceuticals Inc CRYSTALLINE FORMS OF THERAPEUTIC COMPOUNDS and USES THEREOF.
CN104529894B (en) * 2015-01-15 2017-02-22 成都丽凯手性技术有限公司 Quinolinone derivative and preparation method thereof
JP2019533641A (en) 2016-09-08 2019-11-21 カラ ファーマシューティカルズ インコーポレイテッド Crystalline forms of therapeutic compounds and uses thereof
WO2018048747A1 (en) 2016-09-08 2018-03-15 Kala Pharmaceuticals, Inc. Crystalline forms of therapeutic compounds and uses thereof
EP3509423A4 (en) 2016-09-08 2020-05-13 Kala Pharmaceuticals, Inc. Crystalline forms of therapeutic compounds and uses thereof
CN108610293B (en) * 2018-06-15 2020-08-04 南京工业大学 Method for preparing dorvitinib intermediate by adopting microchannel reaction device
AU2019315444A1 (en) * 2018-07-31 2021-02-11 The Trustees Of Princeton University Tetrahydroquinolino derivatives for the treatment of metastatic and chemoresistant cancers
CN117897370A (en) 2021-04-22 2024-04-16 凯悦施那有限公司 Heterocyclic compounds and their use

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000000481A1 (en) * 1998-06-29 2000-01-06 Du Pont Pharmaceuticals Company Cyclic carbamates and isoxazolidines as iib/iiia antagonists
WO2002022598A1 (en) * 2000-09-11 2002-03-21 Chiron Corporation Quinolinone derivatives as tyrosine kinase inhibitors
WO2003087095A1 (en) * 2002-04-05 2003-10-23 Chiron Corporation Quinolinone derivatives
WO2004018419A2 (en) * 2002-08-23 2004-03-04 Chiron Corporation Benzimidazole quinolinones and uses thereof
WO2004030620A2 (en) * 2002-09-30 2004-04-15 Bristol-Myers Squibb Company Novel tyrosine kinase inhibitors
WO2004043389A2 (en) * 2002-11-13 2004-05-27 Chiron Corporation Methods of treating cancer and related methods
WO2004087153A2 (en) * 2003-03-28 2004-10-14 Chiron Corporation Use of organic compounds for immunopotentiation
WO2005046589A2 (en) * 2003-11-07 2005-05-26 Chiron Corporation Pharmaceutically acceptable salts of quinolinone compounds having improved pharmaceutical properties
WO2005053692A1 (en) * 2003-12-01 2005-06-16 The Scripps Research Institute Advanced quinolinone based protein kinase inhibitors

Family Cites Families (73)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3663606A (en) * 1966-06-21 1972-05-16 Mitsui Toatsu Chemicals Organic imino-compounds
DE2363459A1 (en) 1973-12-20 1975-06-26 Basf Ag 7-Amino-quinoline cpds. - for use as optical brighteners, dyes, colour formers in copying and in printing pastes
US4659657A (en) * 1982-12-24 1987-04-21 Bayer Aktiengesellschaft Chromogenic and fluorogenic esters for photometric or fluorimetric determination of phosphatases or sulphatases
DE3248043A1 (en) 1982-12-24 1984-06-28 Bayer Ag, 5090 Leverkusen Fluorogenic phosphoric esters, process for their preparation, and process and composition for the detection and fluorometric determination of phosphates
DE3634066A1 (en) * 1986-10-07 1988-04-21 Boehringer Mannheim Gmbh NEW 5-ALKYLBENZIMIDAZOLES, PROCESS FOR THEIR PRODUCTION AND MEDICINAL PRODUCTS
US5073492A (en) * 1987-01-09 1991-12-17 The Johns Hopkins University Synergistic composition for endothelial cell growth
JPH07121937B2 (en) 1987-03-18 1995-12-25 大塚製薬株式会社 Carbostyril derivative
JPH0699497B2 (en) 1987-04-16 1994-12-07 富士写真フイルム株式会社 Photopolymerizable composition
GB8709448D0 (en) 1987-04-21 1987-05-28 Pfizer Ltd Heterobicyclic quinoline derivatives
JPH02229165A (en) 1989-03-02 1990-09-11 Otsuka Pharmaceut Co Ltd Carbostyril derivative
DE3932953A1 (en) * 1989-10-03 1991-04-11 Boehringer Mannheim Gmbh NEW 2-BICYCLO-BENZIMIDAZOLES, METHOD FOR THEIR PRODUCTION AND MEDICINAL PRODUCTS CONTAINING THESE COMPOUNDS
US5151360A (en) * 1990-12-31 1992-09-29 Biomembrane Institute Effect of n,n,n-trimethylsphingosine on protein kinase-c activity, melanoma cell growth in vitro, metastatic potential in vivo and human platelet aggregation
GB9107742D0 (en) 1991-04-11 1991-05-29 Rhone Poulenc Agriculture New compositions of matter
GB9108369D0 (en) 1991-04-18 1991-06-05 Rhone Poulenc Agriculture Compositions of matter
GB9108547D0 (en) 1991-04-22 1991-06-05 Fujisawa Pharmaceutical Co Quinoline derivatives
WO1992020642A1 (en) 1991-05-10 1992-11-26 Rhone-Poulenc Rorer International (Holdings) Inc. Bis mono-and bicyclic aryl and heteroaryl compounds which inhibit egf and/or pdgf receptor tyrosine kinase
US5480883A (en) * 1991-05-10 1996-01-02 Rhone-Poulenc Rorer Pharmaceuticals Inc. Bis mono- and bicyclic aryl and heteroaryl compounds which inhibit EGF and/or PDGF receptor tyrosine kinase
USRE37650E1 (en) * 1991-05-10 2002-04-09 Aventis Pharmacetical Products, Inc. Aryl and heteroaryl quinazoline compounds which inhibit CSF-1R receptor tyrosine kinase
US5710158A (en) * 1991-05-10 1998-01-20 Rhone-Poulenc Rorer Pharmaceuticals Inc. Aryl and heteroaryl quinazoline compounds which inhibit EGF and/or PDGF receptor tyrosine kinase
US5856115A (en) * 1991-05-24 1999-01-05 Fred Hutchinson Cancer Research Center Assay for identification therapeutic agents
CA2452130A1 (en) * 1992-03-05 1993-09-16 Francis J. Burrows Methods and compositions for targeting the vasculature of solid tumors
JP3142378B2 (en) 1992-06-22 2001-03-07 ティーディーケイ株式会社 Organic EL device
WO1994005333A1 (en) * 1992-09-02 1994-03-17 Isis Pharmaceuticals, Inc. Oligonucleotide modulation of cell adhesion
US5330992A (en) * 1992-10-23 1994-07-19 Sterling Winthrop Inc. 1-cyclopropyl-4-pyridyl-quinolinones
SE9203318D0 (en) 1992-11-06 1992-11-06 Kabi Pharmacia Ab NOVEL 3,3-DIPHENYL PROPYLAMINES, THEIR USE AND PREPARATION
US5981569A (en) * 1992-11-13 1999-11-09 Yissum Research Development Company Of The Hebrew University Of Jerusalem Substituted phenylacrylonitrile compounds and compositions thereof for the treatment of disease
US5763441A (en) * 1992-11-13 1998-06-09 Sugen, Inc. Compounds for the treatment of disorders related to vasculogenesis and/or angiogenesis
US5792771A (en) * 1992-11-13 1998-08-11 Sugen, Inc. Quinazoline compounds and compositions thereof for the treatment of disease
JPH0743896A (en) 1993-07-28 1995-02-14 Toyobo Co Ltd Photopolymerizable composition
US5498608A (en) 1994-01-07 1996-03-12 Salix Pharmaceuticals Use of 2-hydroxy-5-phenylazobenzoic acid derivatives as colon cancer chemopreventative and chemotherapeutic agents
AU5881394A (en) 1994-01-08 1995-08-01 Rhone-Poulenc Agriculture Limited Benzimidazolyl quinoline-3-carboxylate derivatives, intermediates thereto, and their use as herbicides
JPH0829973A (en) 1994-07-11 1996-02-02 Toyobo Co Ltd Photopolymerized composition
JP3441246B2 (en) * 1995-06-07 2003-08-25 富士写真フイルム株式会社 Photopolymerizable composition
GB9514265D0 (en) 1995-07-13 1995-09-13 Wellcome Found Hetrocyclic compounds
WO1997021436A1 (en) 1995-12-12 1997-06-19 Merck & Co., Inc. New use for losartan
GB9624482D0 (en) * 1995-12-18 1997-01-15 Zeneca Phaema S A Chemical compounds
EP0888310B1 (en) 1996-03-15 2005-09-07 AstraZeneca AB Cinnoline derivatives and use as medicine
DE19610723A1 (en) 1996-03-19 1997-09-25 Bayer Ag Electroluminescent devices using glare systems
US5942385A (en) * 1996-03-21 1999-08-24 Sugen, Inc. Method for molecular diagnosis of tumor angiogenesis and metastasis
EP2223920A3 (en) * 1996-06-19 2011-09-28 Aventis Pharma Limited Substituted azabicyclic compounds
CA2258822A1 (en) 1996-06-20 1997-12-24 Sean Kerwin Compounds and methods for providing pharmacologically active preparations and uses thereof
ATE300521T1 (en) 1996-09-25 2005-08-15 Astrazeneca Ab QUINOLINE DERIVATIVES THAT DELAY THE EFFECT OF GROWTH FACTORS LIKE VEGF
US6111110A (en) * 1996-10-30 2000-08-29 Eli Lilly And Company Synthesis of benzo[f]quinolinones
US6245760B1 (en) 1997-05-28 2001-06-12 Aventis Pharmaceuticals Products, Inc Quinoline and quinoxaline compounds which inhibit platelet-derived growth factor and/or p56lck tyrosine kinases
DE69807222T2 (en) 1997-06-02 2003-04-17 Janssen Pharmaceutica N.V., Beerse (IMIDAZOL-5-YL) METHYL-2-QUINOLINONE DERIVATIVES AS INHIBITORS OF PROLIFERATION OF SMOOTH MUSCLE CELLS
GB9716557D0 (en) * 1997-08-06 1997-10-08 Glaxo Group Ltd Benzylidene-1,3-dihydro-indol-2-one derivatives having anti-cancer activity
WO1999010349A1 (en) 1997-08-22 1999-03-04 Zeneca Limited Oxindolylquinazoline derivatives as angiogenesis inhibitors
EP1017682A4 (en) 1997-09-26 2000-11-08 Merck & Co Inc Novel angiogenesis inhibitors
DE19756235A1 (en) * 1997-12-17 1999-07-01 Klinge Co Chem Pharm Fab New piperidinyl-substituted pyridylalkane alkene and alkane carboxylic acid amides
AU3363599A (en) 1998-03-26 1999-10-18 Max-Planck Institut Fur Biochemie Heterocyclic families of compounds for the modulation of tyrosine protein kinase
JP2002509928A (en) 1998-03-31 2002-04-02 ワーナー−ランバート・カンパニー Quinolones as serine protease inhibitors
CA2328893A1 (en) 1998-05-20 1999-11-25 Kyowa Hakko Kogyo Co., Ltd. Vegf activity inhibitors
JP4533534B2 (en) 1998-06-19 2010-09-01 ノバルティス バクシンズ アンド ダイアグノスティックス,インコーポレーテッド Inhibitor of glycogen synthase kinase 3
US6174912B1 (en) * 1998-08-21 2001-01-16 Dupont Pharmaceuticals Company Nitrogen substituted imidazo[4,5-C]pyrazoles as corticotropin releasing hormone antagonists
DE19841985A1 (en) 1998-09-03 2000-03-09 Schering Ag New heterocyclic alkanesulfonic and alkane carboxylic acid derivatives are VEGF receptor blockers useful in treatment of e.g. psoriasis, rheumatoid arthritis, stroke, tumors and endometriosis
US20030087854A1 (en) * 2001-09-10 2003-05-08 Isis Pharmaceuticals Inc. Antisense modulation of fibroblast growth factor receptor 3 expression
JP4707240B2 (en) * 1999-05-05 2011-06-22 アベンティス・フアーマ・リミテッド Urea as a cell adhesion regulator
KR100298572B1 (en) * 1999-08-19 2001-09-22 박찬구 The method for preparing 4-nitrodiphenylamine and 4-nitrosodiphenylamine from carbanilide
ES2235970T3 (en) * 1999-10-19 2005-07-16 MERCK &amp; CO. INC. THYROSINE KINASE INHIBITORS.
AU778588B2 (en) * 1999-10-19 2004-12-09 Merck Sharp & Dohme Corp. Tyrosine kinase inhibitors
US6420382B2 (en) * 2000-02-25 2002-07-16 Merck & Co., Inc. Tyrosine kinase inhibitors
US6313138B1 (en) * 2000-02-25 2001-11-06 Merck & Co., Inc. Tyrosine kinase inhibitors
WO2002020500A2 (en) * 2000-09-01 2002-03-14 Icos Corporation Materials and methods to potentiate cancer treatment
ATE448226T1 (en) * 2000-09-01 2009-11-15 Novartis Vaccines & Diagnostic AZA HETEROCYCLIC DERIVATIVES AND THEIR THERAPEUTIC USE
EP1401831A1 (en) * 2001-07-03 2004-03-31 Chiron Corporation Indazole benzimidazole compounds as tyrosine and serine/threonine kinase inhibitors
US20040208844A1 (en) * 2001-08-01 2004-10-21 Francis Ignatious Products and drug delivery vehicles
US20030083286A1 (en) * 2001-08-22 2003-05-01 Ching-Leou Teng Bioadhesive compositions and methods for enhanced intestinal drug absorption
US20030159702A1 (en) * 2002-01-21 2003-08-28 Lindell Katarina E.A. Formulation and use manufacture thereof
US7825132B2 (en) * 2002-08-23 2010-11-02 Novartis Vaccines And Diagnostics, Inc. Inhibition of FGFR3 and treatment of multiple myeloma
US20050256157A1 (en) * 2002-08-23 2005-11-17 Chiron Corporation Combination therapy with CHK1 inhibitors
US6774327B1 (en) * 2003-09-24 2004-08-10 Agilent Technologies, Inc. Hermetic seals for electronic components
WO2005037306A1 (en) 2003-10-17 2005-04-28 Novo Nordisk A/S Combination therapy
MX2007014612A (en) * 2005-05-18 2008-01-17 M & G Polimeri Italia Spa Polyester composition.

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000000481A1 (en) * 1998-06-29 2000-01-06 Du Pont Pharmaceuticals Company Cyclic carbamates and isoxazolidines as iib/iiia antagonists
WO2002022598A1 (en) * 2000-09-11 2002-03-21 Chiron Corporation Quinolinone derivatives as tyrosine kinase inhibitors
WO2003087095A1 (en) * 2002-04-05 2003-10-23 Chiron Corporation Quinolinone derivatives
WO2004018419A2 (en) * 2002-08-23 2004-03-04 Chiron Corporation Benzimidazole quinolinones and uses thereof
WO2004030620A2 (en) * 2002-09-30 2004-04-15 Bristol-Myers Squibb Company Novel tyrosine kinase inhibitors
WO2004043389A2 (en) * 2002-11-13 2004-05-27 Chiron Corporation Methods of treating cancer and related methods
WO2004087153A2 (en) * 2003-03-28 2004-10-14 Chiron Corporation Use of organic compounds for immunopotentiation
WO2005046589A2 (en) * 2003-11-07 2005-05-26 Chiron Corporation Pharmaceutically acceptable salts of quinolinone compounds having improved pharmaceutical properties
WO2005053692A1 (en) * 2003-12-01 2005-06-16 The Scripps Research Institute Advanced quinolinone based protein kinase inhibitors

Cited By (43)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9012451B2 (en) 2002-09-04 2015-04-21 Novartis Ag Aza-bicycloalkyl ethers and their use as ALPHA7-nachr agonists
US8236803B2 (en) 2002-09-04 2012-08-07 Novartis Ag Aza-bicycloalkyl ethers and their use as alpha7-nAChR agonists
US9567343B2 (en) 2002-09-04 2017-02-14 Novartis Ag Aza-bicyloalkyl ethers and their use as alpha7-nachr agonists
US9849117B2 (en) 2002-09-04 2017-12-26 Novartis Ag Aza-bicycloalkyl ethers and their use as alpha7-nachr agonists
US9051278B2 (en) 2003-07-22 2015-06-09 Astex Therapeutics, Ltd. 3,4-disubstituted 1H-pyrazole compounds and their use as cyclin dependent kinase and glycogen synthase kinase-3 modulators
US8779147B2 (en) 2003-07-22 2014-07-15 Astex Therapeutics, Ltd. 3,4-disubstituted 1H-pyrazole compounds and their use as cyclin dependent kinase and glycogen synthase kinase-3 modulators
US9475811B2 (en) 2004-06-18 2016-10-25 Novartis Ag 1-aza-bicyclo[3.3.1]nonanes
US8933090B2 (en) 2004-06-18 2015-01-13 Novartis Ag 1-aza-bicyclo[3.3.1]nonanes
US8609662B2 (en) 2004-07-14 2013-12-17 Novartis Ag 3-(heteroaryl-oxy)-2-alkyl-1-aza-bicycloalkyl derivatives as alpha. 7-nachr ligands for the treatment of CNS diseases
US9657010B2 (en) 2004-07-14 2017-05-23 Novartis Ag Substituted quinuclidines as alpha 7-nicotinic acetylcholine receptor activity modulators
WO2006077428A1 (en) * 2005-01-21 2006-07-27 Astex Therapeutics Limited Pharmaceutical compounds
US8293767B2 (en) 2005-01-21 2012-10-23 Astex Therapeutics Limited 4-(2,6-dichloro-benzoylamino)-1H-pyrazole-3-carboxylic acid piperidin-4-ylamide acid addition salts as kinase inhibitors
US8404718B2 (en) 2005-01-21 2013-03-26 Astex Therapeutics Limited Combinations of pyrazole kinase inhibitors
EP2301546A1 (en) * 2005-01-27 2011-03-30 Novartis Vaccines and Diagnostics, Inc. Treatment of metastasized tumors
JP2008528617A (en) * 2005-01-27 2008-07-31 ノバルティス ヴァクシンズ アンド ダイアグノスティクス インコーポレイテッド Treatment of tumors that have metastasized
EP1845990A4 (en) * 2005-01-27 2008-04-30 Novartis Vaccines & Diagnostic Treatment of metastasized tumors
EP1845990A2 (en) * 2005-01-27 2007-10-24 Novartis Vaccines and Diagnostics, Inc. Treatment of metastasized tumors
EP2270000A1 (en) * 2005-05-23 2011-01-05 Novartis AG Crystalline and other forms of 4-amino-5-fluoro-3-[6-(4-methylpiperazin-1-yl)-1h-benzimidazol-2-yl]-1h-quinolin-2-one lactic acid salts
EP2270000B1 (en) 2005-05-23 2015-07-29 Novartis AG Crystalline and other forms of 4-amino-5-fluoro-3-[6-(4-methylpiperazin-1-yl)-1H-benzimidazol-2-yl]-1H-quinolin-2-one lactic acid salts
WO2006127926A2 (en) * 2005-05-23 2006-11-30 Novartis Ag Crystalline and other forms of 4-amino-5-fluoro-3-[6-(4-methylpiperazin-1-yl)-1h-benzimidazol-2-yl]-1h-quinolin-2-one lactic acid salts
WO2006127926A3 (en) * 2005-05-23 2007-01-18 Novartis Ag Crystalline and other forms of 4-amino-5-fluoro-3-[6-(4-methylpiperazin-1-yl)-1h-benzimidazol-2-yl]-1h-quinolin-2-one lactic acid salts
JP2013224299A (en) * 2005-05-23 2013-10-31 Novartis Ag Crystal and other forms of 4-amino-5- fluoro-3-[6-(4-methylpiperazin-1-yl)-1h -benzimidazol-2-yl]-1h-quinolin-2-one lactate
US8173667B2 (en) 2005-10-21 2012-05-08 Novartis Ag 1-aza-bicycloalkyl derivatives
US8759346B2 (en) 2005-12-16 2014-06-24 Novartis Ag Organic compounds
US8637517B2 (en) 2005-12-16 2014-01-28 Novartis Ag Organic compounds
US8048885B2 (en) 2005-12-16 2011-11-01 Novartis Ag Organic compounds
US9206181B2 (en) 2005-12-16 2015-12-08 Novartis Ag 1-aza-bicyclo[3.3.1] non-4-yl)-[5-(1H-indol-5-yl)-heteroaryl]-amines as cholinergic ligands of the n-AChR for the treatment of psychotic and neurodegenerative disorders
WO2007077961A3 (en) * 2005-12-28 2007-11-22 Takeda Pharmaceutical Fused heterocyclic compounds and their use as mineralocorticoid receptor ligands
WO2007077961A2 (en) * 2005-12-28 2007-07-12 Takeda Pharmaceutical Company Limited Fused heterocyclic compounds and their use as mineralocorticoid receptor ligands
US8258154B2 (en) 2006-07-07 2012-09-04 Kalypsys Inc. Bicyclic heteroaryl inhibitors of PDE4
US8080563B2 (en) 2006-07-07 2011-12-20 Kalypsys Bicyclic heteroaryl inhibitors of PDE4
US8138205B2 (en) 2006-07-07 2012-03-20 Kalypsys, Inc. Heteroarylalkoxy-substituted quinolone inhibitors of PDE4
US9078887B2 (en) 2006-07-07 2015-07-14 Kalypsys, Inc. Bicyclic heteroaryl inhibitors of PDE4
JP2011515370A (en) * 2008-03-19 2011-05-19 ノバルティス アーゲー Crystal form of 4-amino-5-fluoro-3- [5- (4-methylpiperazin-1-yl) -1H-benzimidazol-2-yl] quinolin-2 (1H) -one lactate and two solvents Japanese style
US8492374B2 (en) 2009-04-29 2013-07-23 Industrial Technology Research Institute Azaazulene compounds
US8741903B2 (en) * 2010-04-16 2014-06-03 Novartis Ag Organic compound for use in the treatment of hepatocellular cancer (HCC)
AU2011239999B2 (en) * 2010-04-16 2014-04-03 Novartis Ag Organic compound for use in the treatment of liver cancer
US20130123272A1 (en) * 2010-04-16 2013-05-16 Novartis Ag Organic compound for use in the treatment of liver cancer
WO2011128403A1 (en) * 2010-04-16 2011-10-20 Novartis Ag Organic compound for use in the treatment of liver cancer
AU2018246382B2 (en) * 2017-03-31 2022-06-16 Vivoryon Therapeutics N.V. Novel inhibitors
WO2019001419A1 (en) * 2017-06-27 2019-01-03 Janssen Pharmaceutica Nv New quinolinone compounds
US11384075B2 (en) 2017-06-27 2022-07-12 Janssen Pharmaceutica Nv Quinolinone compounds
AU2018291687B2 (en) * 2017-06-27 2022-07-14 Janssen Pharmaceutica Nv New quinolinone compounds

Also Published As

Publication number Publication date
AU2005216904B2 (en) 2010-11-25
JP2007523185A (en) 2007-08-16
CN1960731B (en) 2011-12-07
US20050239825A1 (en) 2005-10-27
EP1718306A2 (en) 2006-11-08
US7875624B2 (en) 2011-01-25
RU2377988C2 (en) 2010-01-10
RU2006133536A (en) 2008-03-27
CA2556872A1 (en) 2005-09-09
BRPI0507891A (en) 2007-07-24
IL177574A0 (en) 2006-12-10
WO2005082340A3 (en) 2006-05-04
CA2556872C (en) 2015-05-12
CN1960731A (en) 2007-05-09
AU2005216904A1 (en) 2005-09-09
JP5019884B2 (en) 2012-09-05

Similar Documents

Publication Publication Date Title
EP1718306A2 (en) Modulation of inflammatory and metastatic processes
US10988456B2 (en) O-aminoheteroaryl alkynyl-containing compound, preparation method therefor, and use thereof
US20050137399A1 (en) Methods for synthesizing quinolinone compounds
US20040102455A1 (en) Method of inhibiting kinases
US8299081B2 (en) Methods for treating drug resistant cancer
JP2022515293A (en) Chromoly esters and their use
EA006711B1 (en) Quinolinone derivatives as tyrosine kinase inhibitors
JPH07503018A (en) Pyridyl-substituted imidazole
EA010393B1 (en) Quinolinone derivatives
JP7012289B2 (en) Benzoylglycine derivatives and methods for their preparation and use
JP2020143099A (en) COMBINATION THERAPY OF INHIBITORS OF C-C CHEMOKINE RECEPTOR 9 (CCR9) AND ANTI-α4β7 INTEGRIN BLOCKING ANTIBODIES
JP2020536853A (en) Small molecule inhibition of the transcription factor SALL4 and its use
US20240067635A1 (en) Cromolyn derivatives and uses thereof
JPH05503705A (en) 5-oxygenated-2,4,6-triaminopyrimidines
MXPA06009470A (en) Modulation of inflammatory and metastatic processes
JP2023142282A (en) Medical composition for treating disease that generate filamentation
WO2024106529A1 (en) Novel compound having anti-obesity activity
KR101051078B1 (en) 2,4-disubstituted-5-aminocarbonyl-1,3-thiazole derivatives for the treatment of inflammation-related diseases, methods for their preparation and agents for the treatment of inflammation-related diseases caused by SPC receptor activity containing them as active ingredients
WO2023070076A1 (en) Compounds for cancers driven by braf mutation
WO2001001981A1 (en) Vegf receptor antagonists
JP2629670B2 (en) Anti-rheumatic drug
JP2022505588A (en) Composition for the prevention or treatment of Sjogren&#39;s syndrome
JP2003063966A (en) Interleukin 6-production inhibitor
JPH03148263A (en) Phenoxypropylamine derivative or salt thereof and antiulcer agent containing the same
AU2002226197A1 (en) Methods of inhibiting kinases

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BW BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE EG ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NA NI NO NZ OM PG PH PL PT RO RU SC SD SE SG SK SL SM SY TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): BW GH GM KE LS MW MZ NA SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LT LU MC NL PL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
WWE Wipo information: entry into national phase

Ref document number: 2006554253

Country of ref document: JP

Ref document number: 2005216904

Country of ref document: AU

Ref document number: 177574

Country of ref document: IL

WWE Wipo information: entry into national phase

Ref document number: 2556872

Country of ref document: CA

Ref document number: PA/a/2006/009470

Country of ref document: MX

NENP Non-entry into the national phase

Ref country code: DE

WWW Wipo information: withdrawn in national office

Ref document number: DE

WWE Wipo information: entry into national phase

Ref document number: 2005723338

Country of ref document: EP

ENP Entry into the national phase

Ref document number: 2005216904

Country of ref document: AU

Date of ref document: 20050218

Kind code of ref document: A

WWP Wipo information: published in national office

Ref document number: 2005216904

Country of ref document: AU

WWE Wipo information: entry into national phase

Ref document number: 2006133536

Country of ref document: RU

Ref document number: 2736/KOLNP/2006

Country of ref document: IN

WWE Wipo information: entry into national phase

Ref document number: 200580009523.1

Country of ref document: CN

WWP Wipo information: published in national office

Ref document number: 2005723338

Country of ref document: EP

ENP Entry into the national phase

Ref document number: PI0507891

Country of ref document: BR