Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
  • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
  • C12Q1/48—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase
  • C12Q1/485—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase involving kinase
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
  • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
  • C07D471/04—Ortho-condensed systems
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D513/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
  • C07D513/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains two hetero rings
  • C07D513/04—Ortho-condensed systems
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
  • G01N2333/90—Enzymes; Proenzymes
  • G01N2333/91—Transferases (2.)
  • G01N2333/912—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
  • G01N2333/91205—Phosphotransferases in general
  • G01N2333/9121—Phosphotransferases in general with an alcohol group as acceptor (2.7.1), e.g. general tyrosine, serine or threonine kinases
  • G01N2333/91215—Phosphotransferases in general with an alcohol group as acceptor (2.7.1), e.g. general tyrosine, serine or threonine kinases with a definite EC number (2.7.1.-)
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N2500/00—Screening for compounds of potential therapeutic value
  • G01N2500/04—Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A)

Definitions

• the invention relates in part to molecules having certain biological activities that include, but are not limited to, inhibiting cell proliferation, and modulating serine-threonine protein kinase activity
• Molecules of the invention can modulate casein kinase (CK) activity (e g , CK2 activity)
• CK casein kinase
• the invention also relates in part to methods for using such molecules
• the present invention in part provides chemical compounds having certain biological activities that include, but are not limited to, inhibiting cell proliferation, inhibiting angiogenesis, modulating protein kinase activity and modulating polymerase activity
• Certain molecules can modulate casein kinase 2 (CK2) activ ity and can affect biological functions that include but are not limited to, inhibiting gamma phosphate transfer from ATP to a protein or peptide substrate, inhibiting angiogenesis, inhibiting cell proliferation and inducing cell apoptosis, for example
• the present invention also in part prov ides methods for preparing novel chemical compounds, and analogs thereof, and methods of using the foregoing
• group labeled ⁇ represents a 5-6 membered aromatic or heteroaromatic ring fused onto the ring containing Q 1 , wherein ⁇ is a 6-membered aryl ⁇ ng optionally containing one or more nitrogen atoms as ⁇ ng members, or a five membered aryl ⁇ ng selected from thiophene and thiazole,
• the invention also includes the pharmaceutically acceptable salts of compounds of formula (A).
• Formula (A) represents a fused tricyclic ring system which is linked through Q 1 to a group R 10 , which is further described below.
• the invention provides compounds of Formula (I):
• each Z 1 , Z 2 , Z 3 , and Z 4 is N, CH, or CR 3 , provided that none, one or two of Z 1 , Z 2 , Z 3 , and Z 4 are N; and further provided that one or more of Z 1 , Z 2 , Z 3 , and Z 4 is CR 3 ;
• each R 3 is independently an optionally substituted C1-C8 alkyl, C2-C8 heteroalkyl, C2- C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C12 heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkyl group, or each R 3 is independently halo, CF 3 , OR, NR 2
• Z 5 is CR 6A or N; each of R 6A , R 6B and R 6C is independently H or an optionally substituted C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C12 heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkyl group, or each of R 6A , R 6B and R 6C can be halo, CF 3 , OR, NR 2 , NROR, NRNR 2 , SR, SOR, SO 2 R, SO 2 NR 2 , NRSO 2 R, NRCONR 2 , NRCOOR, NRCOR, CN, COOR, CONR 2 , 0OCR, COR, or
• R 10 is an optionally substituted five-membered or six-membered carbocyclic or heterocyclic aromatic ring.
• Z 5 is N or CR 6A , such that the ring containing Z 5 is independently an optionally substituted pyridine or pyrimidine ring.
• one or more ring nitrogen atoms within the ring containing Z 5 may be arranged as follows:
• R 6A , R 6B , and R 6C is independently selected from the R 6 substituents defined above with respect to compounds of Formula (I).
• each of Z 1 , Z 2 , Z 3 , and Z 4 is N, CH, or CR 3 , provided that none, one or two of Z 1 , Z 2 , Z 3 , and Z 4 are N; and further provided that one or more of Z 1 , Z 2 , Z 3 , and Z 4 is CR 3 , where each R 3 is independently selected from the R 3 substituents defined above with respect to compounds of Formula (I).
• At least one of Z 1 , Z 2 , Z 3 , and Z 4 is a nitrogen atom.
• One or more ring nitrogen atoms can be positioned in the ring containing Z 1 , Z 2 , Z 3 , and Z 4 such that each ring is independently an optionally substituted pyridine, pyrimidine or pyridazine ring.
• each of Z 1 , Z 2 , Z 3 , and Z 4 is CR 3 , where each R 3 is independently selected from the R 3 substituents defined above with respect to compounds of Formula (I).
• one or more of Z 1 , Z 2 , Z 3 , and Z 4 is CR 3 , and the others are CH.
• one of Z 1 , Z 2 , Z 3 , and Z 4 is CR 3 , and the others are CH.
• Z 2 is CR 3 and Z 1 , Z 3 , and Z 4 are CH.
• At least one R 3 substituent is a polar substituent, such as a carboxylic acid or a salt, an ester or a bioisostere thereof.
• at least one R 3 is a carboxylic acid-containing substituent or a carboxylate bioisostere, or a salt or ester thereof, for example.
• at least one R 3 is a carboxylic acid, carboxylate salt, ester, carboxamide, tetrazole or other carboxy bioisostere.
• at least one R 3 is a carboxylic acid-containing substituent or a salt thereof. This polar substituent is sometimes referred to herein as R 3P .
• polar substituent refers to any substituent having an electric dipole, and optionally a dipole moment (e.g., an asymmetrical polar substituent has a dipole moment and a symmetrical polar substituent does not have a dipole moment).
• Polar substituents include substituents that accept or donate a hydrogen bond, and groups that would carry at least a partial positive or negative charge in aqueous solution at physiological pH levels.
• a polar substituent is one that can accept or donate electrons in a non- covalent hydrogen bond with another chemical moiety.
• a polar substituent is selected from a carboxy, a carboxy bioisostere or other acid-derived moiety that exists predominately as an anion at a pH of about 7 to 8.
• Other polar substituents include, but are not limited to, groups containing an OH or NH, an ether oxygen, an amine nitrogen, an oxidized sulfur or nitrogen, a carbonyl, a nitrile, and a nitrogen-containing or oxygen-containing heterocyclic ring whether aromatic or non-aromatic.
• the polar substituent represented by R 3 is a carboxylate or a carboxylate bioisostere.
• Carboxylate bioisostere or “carboxy bioisostere” as used herein refers to a moiety that is expected to be negatively charged to a substantial degree at physiological pH.
• the carboxylate bioisostere is a moiety selected from the group consisting of:
• each R 7 is independently H or an optionally substituted member selected from the group consisting of Cj. io alkyl, C M0 alkenyl, C 2 .io heteroalkyl, C 3 . 8 carbocyclic ring, and C 3 . 8 heterocyclic ring optionally fused to an additional optionally substituted carbocyclic or heterocyclic ring; or R 7 is a C MO alkyl, C 2 - I o alkenyl, or C 2 . io heteroalkyl substituted with an optionally substituted C 3 . 8 carbocyclic ring or C 3 .g heterocyclic ring.
• the polar substituent is selected from the group consisting of carboxylic acid, carboxylic ester, carboxamide, tetrazole, triazole, carboxymethanesulfonamide, oxadiazole, oxathiadiazole, thiazole, aminothiazole and hydroxythiazole.
• At least one R 3 present is a carboxylic acid or a salt, or ester or a bioisostere thereof.
• at least one R 3 present is a carboxylic acid- containing substituent or a salt, ester or bioisostere thereof.
• the R 3 substituent may be a Cl-ClO alkyl or C2-C10 alkenyl linked to a carboxylic acid (or salt, ester or bioisostere thereof), for example, and in some embodiments, the R 3 substituent is not NHCOOCH 2 CH 3 .
• a polar substituent may be at any position on the ring containing Z'-Z 4 in Formula (I), and the ring may include one, two, three or four polar substituents.
• at least one of Z'-Z 4 may be CR 3 and one of the R 3 substituents may be a polar substituent (e.g., a carboxylate or carboxylic acid ester, or a tetrazole) arranged at any one of the positions in the ring containing Z'-Z 4 :
• one of Z'-Z 4 is CR 3 and the others of Z 1 -Z 4 are CH, where R 3 is a polar substituent (i.e., R 3P ), for example, a carboxylate or carboxylic acid, ester, or amide, or a tetrazole, arranged at any one of the positions in the ring containing Z 1 -Z 4 :
• R 3 is a polar substituent (i.e., R 3P ), for example, a carboxylate or carboxylic acid, ester, or amide, or a tetrazole, arranged at any one of the positions in the ring containing Z 1 -Z 4 :
• the polar substituent represented by R 3 in some embodiments is a carboxy (e.g., COOH), carboxyalkyl (e.g., carboxymethyl), tetrazole or amide (e.g., -CONH 2, .CONHc-Pr) substituent.
• R 3 represents a carboxylate bioisostere.
• R 3 at position Z 2 is a carboxylic acid-containing substituent or a carboxylate bioisostere, or a salt or ester thereof.
• Z 2 is CR 3 , where R 3 is COOH or COOMe.
• it is CONR 2 , where each R is independently H or C M alkyl; sometimes, it is CONH 2 or CONHc-Pr.
• R 10 is an optionally substituted five-membered or six- membered carbocyclic or heterocyclic aromatic ring.
• R 10 is an optionally substituted six-membered carbocyclic or heterocyclic aromatic ring.
• the optionally substituted six-membered carbocyclic or heterocyclic aromatic ring is selected from phenyl, pyridine, pyridazine, pyrimidine, pyrazine, triazine, pyran, and optionally substituted variants of the foregoing.
• R 10 is an optionally substituted phenyl or pyridyl ring ring.
• R 10 is phenyl substituted with one or more halo, CN, C M alkyl, C M alkoxy, NH 2 , or NHAc substitutents.
• R 10 is an optionally substituted five-membered heterocyclic aromatic ring.
• the optionally substituted five-membered heterocyclic aromatic ring is selected from pyrrole, imidazole, pyrazole, triazole, tetrazole, thiophene, thiazole, thiadiazole, isothiazole, furan, oxazole, isoxazole, oxadiazole and optionally substituted variants thereof.
• R 10 is an optionally substituted pyrazole, isoxazole or thiophene; in specific embodiments, it is an N'-methyl-pyrazol-4-yl substituent or an optionally substituted 2- or 3-thiophene substituent.
• optional substituents when present on a five-membered heterocyclic aromatic ring include one or more halo, CN, or C M alkyl groups.
• R 6B , R 6C and R 6A are independently selected from the list defined above with respect to compounds of Formula (I).
• at least one of R 6B and R 6C is H.
• both R 6B and R 6C are H.
• R 6B is thiomethoxy (-SMe) and R 6C is H.
• Z 5 is N, and R 6B is SR, SO 2 R, or NR 2 , where each R is independently H or C 14 alkyl; in some such embodiments, R 6B is SMe, SO 2 Me or NHc-Pr.
• Z 5 is R 6A , and each of R 6A , R 6B and R 6C is H.
• the invention provides compounds of Formula (II):
• each Z 1 , Z 2 , Z 3 , and Z 4 is N, CH, or CR 3 , provided that none, one or two of Z 1 , Z 2 , Z 3 , and Z 4 are N; and further provided that one or more of Z 1 , Z 2 , Z 3 , and Z 4 is CR 3 ;
• each R 3 is independently an optionally substituted C1-C8 alkyl, C2-C8 heteroalkyl, C2- C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C12 heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkyl group, or each R 3 is independently halo, CF 3 , OR
• Z 5 is CR 6A or N; each of R 6A and R 6B is independently H or an optionally substituted C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C12 heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkyl group, or each of R 6A and R 6B can be halo, CF 3 , OR, NR 2 , NROR, NRNR 2 , SR, SOR, SO 2 R, SO 2 NR 2 , NRSO 2 R, NRCONR 2 , NRCOOR, NRCOR, CN, COOR, CONR 2 , OOCR, COR, or NO 2 ; wherein each R is independently H
• R 10 is an optionally substituted five-membered or six-membered carbocyclic or heterocyclic aromatic ring.
• Embodiments described with respect to compounds of Formula (I) for substituents Z 1 , Z 2 , Z 3 , Z 4 , Z 5 , R 6A , R 6B , R 3 and R 10 may also be applied to compounds of Formula (II).
• Z 5 is N or CR 6A , such that the ring containing Z 5 is independently an optionally substituted thiophene or thiazole ring.
• one or more heteroatoms within the ring containing Z 5 may be arranged as follows:
• each of R 6A and R 6B are independently selected from the R 6 substituents defined above with respect to compounds of Formula (II).
• each of Z 1 , Z 2 , Z 3 , and Z 4 is N, CH, or CR 3 , provided that none, one or two of Z 1 , Z 2 , Z 3 , and Z 4 are N; and further provided that one or more of Z 1 , Z 2 , Z 3 , and Z 4 is CR 3 , where each R 3 is independently selected from the R 3 substituents defined above with respect to compounds of Formula (I) or (II).
• At least one of Z'-Z 4 is a nitrogen atom.
• One or more ring nitrogen atoms can be positioned in the ring containing Z'-Z 4 such that each ring is independently an optionally substituted pyridine, pyrimidine or pyridazine ring.
• each of Z 1 , Z 2 , Z 3 and Z 4 is CR 3 , where each R 3 is independently selected from the R 3 substituents defined above with respect to compounds of Formula (II).
• at least one of Z 1 , Z 2 , Z 3 and Z 4 is CR 3 , and the others are CH.
• one of Z 1 , Z 2 , Z 3 and Z 4 is CR 3 , and the others are CH.
• Z 2 is CR 3
• Z 1 , Z 3 and Z 4 are CH.
• At least one R 3 substituent is a polar substituent, such as a carboxylic acid or a salt, an ester or a bioisostere thereof.
• a polar substituent may be at any position on the ring containing Z'-Z 4 , and the ring may include one, two, three or four polar substituents.
• At least one of Z'-Z 4 may be CR 3 and one of the R 3 substituents may be a polar substituent (e.g., a carboxylate or carboxylic acid ester, or a tetrazole) arranged at any one of the positions in the ring, as described for compounds of Formula (I).
• at least one R 3 is a carboxylic acid-containing substituent or a carboxylate bioisostere, or a salt or ester thereof, for example.
• at least one R 3 is a carboxylic acid-containing substituent or a salt thereof.
• R 10 is an optionally substituted five-membered or six- membered carbocyclic or heterocyclic aromatic ring.
• R 10 is an optionally substituted six-membered carbocyclic or heterocyclic aromatic ring.
• the optionally substituted six-membered carbocyclic or heterocyclic aromatic ring is selected from phenyl, pyridine, pyridazine, pyrimidine, pyrazine, triazine, pyran, and optionally substituted variants of the foregoing.
• R 10 is an optionally substituted phenyl ring.
• R 10 is an optionally substituted five-membered heterocyclic aromatic ring.
• the optionally substituted f ⁇ ve- membered heterocyclic aromatic ring is selected from pyrrole, imidazole, pyrazole, triazole, tetrazole, thiophene, thiazole, thiadiazole, isothiazole, furan, oxazole, isoxazole, oxadiazole and optionally substituted variants thereof.
• R 6B and R 6A are independently selected from the list defined above with respect to compounds of Formula (II).
• at least one of R 6A and R 6B is H.
• both R 6A and R 6B are H.
• R 6B is halo or NR 2 .
• Z 5 is CR 6A , where R 6A is H, and R 6B is H or halo.
• Z 5 is N, and R 6B is H or NR 2 .
• each of Z 1 , Z 2 , Z 3 , and Z 4 is N, CH, or CR 3 , provided that none, one or two of Z 1 , Z 2 , Z 3 , and Z 4 are N; and further provided that one or more of Z 1 , Z 2 , Z 3 , and Z 4 is CR 3 , where each R 3 is independently selected from the R 3 substituents defined above with respect to compounds of Formula (I).
• At least one of Z 1 , Z 2 , Z 3 , and Z 4 is a nitrogen atom.
• one or more of Z 1 , Z 2 , Z 3 , and Z 4 is CR 3 , and the others are CH.
• one of Z 1 , Z 2 , Z 3 , and Z 4 is CR 3 , and the others are CH.
• Z 2 is CR 3 and Z 1 , Z 3 , and Z 4 are CH.
• At least one R 3 substituent is a polar substituent, such as a carboxylic acid or a salt, an ester or a bioisostere thereof.
• at least one R 3 is a carboxylic acid-containing substituent or a carboxylate bioisostere, or a salt or ester thereof, for example.
• at least one R 3 is a carboxylic acid-containing substituent or a salt thereof.
• R 3 at position Z 2 is COOH, COOR or CONR 2 , where each R is independently H or C M alkyl.
• R 3 at position Z 2 is COOH, COOMe or CONHc-Pr.
• Z 2 is CR 3 where R 3 is F.
• R 10 is an optionally substituted five-membered heterocyclic aromatic ring or six-membered carbocyclic or heterocyclic aromatic ring.
• R 10 is an optionally substituted phenyl or pyridine ring. In some embodiments, it is an unsubstituted phenyl ring.
• R 10 is phenyl substituted with one or more halo, CN, C M alkyl, C) ⁇ alkoxy, NH 2 , or NHAc substitutents.
• R 10 is an optionally substituted five-membered heterocyclic aromatic ring.
• R 10 is an optionally substituted pyrazole, isoxazole or thiophene ring.
• R 6A , R 6B , and R 6C where present in compounds of Formula (III), (IV), (V) and (VI) is independently selected from the list defined above with respect to compounds of Formula (I).
• at least one of R 6B and R 6C is H.
• both R 6B and R 60 are H.
• R 6B is SR, SO 2 R, or NR 2 , where each R is independently H or C M alkyl; in some such embodiments, R 6B is SMe, SO 2 Me or NHc-Pr.
• R 6A , R 6B and R 6C are H.
• compositions comprising a compound of .
• Pharmaceutical compositions can be utilized in treatments descnbed herein
• identifying a candidate molecule that interacts with a CK2 protein which comprise contacting a composition containing a CK2 protein and a compound desc ⁇ bed herein with a candidate molecule under conditions in which the compound and the protein interact, and determining whether the amount of the compound that interacts with the protein is modulated relative to a control interaction between the compound and the protein without the candidate molecule, whereby a candidate molecule that modulates the amount of the compound interacting with the protein relative to the control interaction is identified as a candidate molecule that interacts with the protein
• the protein is a CK2 protein, such as a CK2 protein comp ⁇ sing the amino acid sequence of SEQ ID NO 1 , 2 or 3 or a substantially identical variant thereof, for example SEO ID NQ 1 (NP 001886, casein kinase II alpha 1 subunit isoform a [Homo sapiensl)
• the protein is in a cell or in a cell-free system.
• the protein, the compound or the molecule in some embodiments is in association with a solid phase.
• the interaction between the compound and the protein is detected via a detectable label, where in some embodiments the protein comprises a detectable label and in certain embodiments the compound comprises a detectable label.
• the interaction between the compound and the protein sometimes is detected without a detectable label.
• Also provided are methods for modulating the activity of a CK2 protein which comprise contacting a system comprising the protein with a compound described herein in an amount effective for modulating the activity of the protein.
• methods for modulating the activity of a Pirn protein or a Fit protein which comprise contacting a system comprising the protein with a compound described herein in an amount effective for modulating the activity of the protein.
• the activity of the protein is inhibited, and sometimes the protein is a CK2 protein, such as a CK2 protein comprising the amino acid sequence of SEQ ID NO: 1, 2 or 3 or a substantially identical variant thereof, for example.
• the system is a cell, and in other embodiments the system is a cell-free system.
• the protein or the compound may be in association with a solid phase in certain embodiments.
• the cells sometimes are in a cell line, such as a cancer cell line (e.g., breast cancer, prostate cancer, pancreatic cancer, lung cancer, hemopoietic cancer, colorectal cancer, skin cancer, ovary cancer cell line), for example.
• the cancer cell line is a breast cancer, prostate cancer or pancreatic cancer cell line.
• the cells sometimes are in a tissue, can be in a subject, at times are in a tumor, and sometimes are in a tumor in a subject.
• the method further comprises inducing cell apoptosis. Cells sometimes are from a subject having macular degeneration.
• the cell proliferative condition is a tumor-associated cancer.
• the cancer sometimes is of the breast, prostate, pancreas, lung, colorectum, skin, or ovary.
• the cell proliferative condition is a non-tumor cancer, such as a hematopoietic cancer, for example.
• the cell proliferative condition is macular degeneration in some embodiments.
• kits for treating cancer or an inflammatory disorder in a subject in need of such treatment comprising: administering to the subject a therapeutically effective amount of a therapeutic agent as described herein; and administering to the subject a molecule that inhibits CK2, Pirn or Rt in an amount that is effective to enhance a desired effect of the therapeutic agent.
• the therapeutic agent sometimes is a compound of formula TA 1 - 1 , TA2, TA3-1, TA4-1 , TA5- 1 or TA6-l as described herein, or a pharmaceutically acceptable salt of one of these compounds.
• the molecule that inhibits CK2, Pirn or FH is a compound of Formula (I), (II), (III), (IV), (V), or (VI) as described herein, or a pharmaceutically acceptable salt thereof.
• the molecule that inhibits CK2, Pirn or Fit is a known compound, or a compound provided herein, or a pharmaceutically acceptable salt of one of these compounds.
• the desired effect of the therapeutic agent that is enhanced by the molecule that inhibits CK2, Pirn or Fit is a reduction in cell proliferation.
• the desired effect of the therapeutic agent that is enhanced by the molecule that inhibits CK2, Pirn or Fit is an increase in apoptosis in at least one type of cell.
• the therapeutic agent in certain embodiments is:
• the therapeutic agent and the molecule that inhibits CK2, Pirn or Fit are administered at substantially the same time.
• the therapeutic agent and molecule that inhibits CK2 sometimes are used concurrently by the subject.
• the therapeutic agent and the molecule that inhibits CK2, Pirn or Fit are combined into one pharmaceutical composition in certain embodiments.
• Some embodiments are directed to a pharmaceutical composition comprising a therapeutic agent of any of formulas TA 1 - 1 , TA2, TA3- 1 , TA 4- 1 , TA5- 1 or TA6 admixed with a molecule that inhibits CK2, Pirn or Fit, or a pharmaceutically acceptable salt thereof.
• the molecule that inhibits CK2, Pirn or Fit is a compound of Formula (I), (II), (III), (IV), (V), or (VI) as described herein, or a pharmaceutically acceptable salt thereof.
• the therapeutic agent is a compound of formula TA2 or a pharmaceutically acceptable salt thereof.
• a therapeutic composition in certain embodiments comprises a therapeutically effective amount of a therapeutic agent of the formula TA2:
• a CK modulator or a pharmaceutically acceptable salt thereof or a specific isomer or mixture of isomers thereof, admixed with an amount of a CK modulator or a pharmaceutically acceptable salt of a CK modulator; and where the amount of the CK modulator or the pharmaceutically acceptable salt of a CK modulator is an amount that is effective to enhance a desired effect of the therapeutic agent.
• compositions comprising a compound described herein and an isolated protein.
• the protein sometimes is a CK2 protein, such as a CK2 protein comprising the amino acid sequence of SEQ ED NO: 1, 2 or 3 or a substantially identical variant thereof, for example.
• the protein is a Pirn protein.
• the protein is a Fit protein.
• Certain compositions comprise a compound described herein in combination with a cell.
• the cell may be from a cell line, such as a cancer cell line. In the latter embodiments, the cancer cell line is sometimes a breast cancer, prostate cancer, pancreatic cancer, lung cancer, hemopoietic cancer, colorectal cancer, skin cancer, ovary cancer cell line.
• Compounds of Formula (I), (II), (III), (IV), (V), and (VI) can exert biological activities that include, but are not limited to, inhibiting cell proliferation, modulating protein kinase activity and modulating polymerase activity.
• Compounds of Formula (I), (II), (III), (IV), (V), and (VI) can modulate CK2 activity, for example. Such compounds therefore can be utilized in multiple applications by a person of ordinary skill in the art.
• compounds described herein may find uses that include, but are not limited to, (i) modulation of protein kinase activity (e.g., CK2 activity), (ii) modulation of cell proliferation, (iii) modulation of apoptosis, and (iv) treatments of cell proliferation related disorders (e.g., administration alone or co-administration with another molecule).
• protein kinase activity e.g., CK2 activity
• modulation of cell proliferation e.g., CK2 activity
• modulation of cell proliferation e.g., CK2 activity
• modulation of cell proliferation e.g., apoptosis
• treatments of cell proliferation related disorders e.g., administration alone or co-administration with another molecule.
• the group takes up two available valences, so the total number of substituents that may be included is reduced according to the number of available valences.
• the compounds of the invention often have ionizable groups so as to be capable of preparation as salts.
• a pharmaceutically acceptable salt may also be used.
• These salts may be acid addition salts involving inorganic or organic acids or the salts may, in the case of acidic forms of the compounds of the invention be prepared from inorganic or organic bases.
• the compounds are prepared or used as pharmaceutically acceptable salts prepared as addition products of pharmaceutically acceptable acids or bases.
• Suitable pharmaceutically acceptable acids and bases are well-known in the art, such as hydrochloric, sulphuric, hydrobromic, acetic, lactic, citric, or tartaric acids for forming acid addition salts, and potassium hydroxide, sodium hydroxide, ammonium hydroxide, caffeine, various amines, and the like for forming basic salts. Methods for preparation of the appropriate salts are well-established in the art.
• the compounds may contain both an acidic and a basic functional group, in which case they may have two ionized groups and yet have no net charge.
• the compounds of the invention contain one or more chiral centers.
• the invention includes each of the isolated stereoisomeric forms as well as mixtures of stereoisomers in varying degrees of chiral purity, including racemic mixtures. It also encompasses the various diastereomers and tautomers that can be formed.
• the compounds of the invention may also exist in more than one tautomeric form; the depiction herein of one tautomer is for convenience only, and is also understood to encompass other tautomers of the form shown.
• alkyl straight-chain, branched-chain and cyclic monovalent hydrocarbyl radicals, and combinations of these, which contain only C and H when they are unsubstituted. Examples include methyl, ethyl, isobutyl, cyclohexyl, cyclopentylethyl, 2-propenyl, 3-butynyl, and the like.
• the total number of carbon atoms in each such group is sometimes described herein, e.g., when the group can contain up to ten carbon atoms it can be represented as 1- 1OC or as Cl-ClO or Cl-10.
• heteroatoms N, O and S typically
• the numbers describing the group though still written as e.g. C1-C6, represent the sum of the number of carbon atoms in the group plus the number of such heteroatoms that are included as replacements for carbon atoms in the backbone of the ring or chain being described.
• the alkyl, alkenyl and alkynyl substituents of the invention contain 1-lOC (alkyl) or 2- 1OC (alkenyl or alkynyl). Preferably they contain 1-8C (alkyl) or 2-8C (alkenyl or alkynyl). Sometimes they contain 1-4C (alkyl) or 2-4C (alkenyl or alkynyl).
• a single group can include more than one type of multiple bond, or more than one multiple bond; such groups are included within the definition of the term "alkenyl” when they contain at least one carbon- carbon double bond, and are included within the term "alkynyl" when they contain at least one carbon-carbon triple bond.
• Alkyl, alkenyl and alkynyl groups are often optionally substituted to the extent that such substitution makes sense chemically.
• Alkyl, alkenyl and alkynyl groups can also be substituted by C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl or C5-C10 heteroaryl, each of which can be substituted by the substituents that are appropriate for the particular group.
• Heteroalkyl “heteroalkenyl”, and “heteroalkynyl” and the like are defined similarly to the corresponding hydrocarbyl (alkyl, alkenyl and alkynyl) groups, but the 'hetero' terms refer to groups that contain 1-3 O, S or N heteroatoms or combinations thereof within the backbone residue; thus at least one carbon atom of a corresponding alkyl, alkenyl, or alkynyl group is replaced by one of the specified heteroatoms to form a heteroalkyl, heteroalkenyl, or heteroalkynyl group.
• heteroforms of alkyl, alkenyl and alkynyl groups are generally the same as for the corresponding hydrocarbyl groups, and the substituents that may be present on the heteroforms are the same as those described above for the hydrocarbyl groups.
• substituents that may be present on the heteroforms are the same as those described above for the hydrocarbyl groups.
• such groups do not include more than two contiguous heteroatoms except where an oxo group is present on N or S as in a nitro or sulfonyl group.
• alkyl as used herein includes cycloalkyl and cycloalkylalkyl groups
• cycloalkyl may be used herein to describe a carbocyclic non-aromatic group that is connected via a ring carbon atom
• cycloalkylalkyl niay be used to describe a carbocyclic non-aromatic group that is connected to the molecule through an alkyl linker.
• heterocyclyl may be used to describe a non-aromatic cyclic group that contains at least one heteroatom as a ring member and that is connected to the molecule via a ring atom, which may be C or N; and “heterocyclylalkyl” may be used to describe such a group that is connected to another molecule through a linker.
• the sizes and substituents that are suitable for the cycloalkyl, cycloalkylalkyl, heterocyclyl, and heterocyclylalkyl groups are the same as those described above for alkyl groups. As used herein, these terms also include rings that contain a double bond or two, as long as the ring is not aromatic.
• acyl encompasses groups comprising an alkyl, alkenyl, alkynyl, aryl or arylalkyl radical attached at one of the two available valence positions of a carbonyl carbon atom
• heteroacyl refers to the corresponding groups wherein at least one carbon other than the carbonyl carbon has been replaced by a heteroatom chosen from N, O and S.
• Acyl and heteroacyl groups are bonded to any group or molecule to which they are attached through the open valence of the carbonyl carbon atom. Typically, they are C1 -C8 acyl groups, which include formyl, acetyl, pivaloyl, and benzoyl, and C2-C8 heteroacyl groups, which include methoxyacetyl, ethoxycarbonyl, and 4-pyridinoyl.
• the hydrocarbyl groups, aryl groups, and heteroforms of such groups that comprise an acyl or heteroacyl group can be substituted with the substituents described herein as generally suitable substituents for each of the corresponding component of the acyl or heteroacyl group.
• Aromatic moiety or "aryl” moiety refers to a monocyclic or fused bicyclic moiety having the well-known characteristics of aromaticity; examples include phenyl and naphthyl.
• heteroaryl refers to such monocyclic or fused bicyclic ring systems which contain as ring members one or more heteroatoms selected from O, S and N. The inclusion of a heteroatom permits aromaticity in 5-membered rings as well as 6-membered rings. Any monocyclic or fused ring bicyclic system which has the characteristics of aromaticity in terms of electron distribution throughout the ring system is included in this definition.
• the ring systems contain 5-12 ring member atoms.
• the monocyclic heteroaryls contain 5-6 ring members, and the bicyclic heteroaryls contain 8-10 ring members.
• Typical heteroaromatic systems include monocyclic C5-C6 aromatic groups such as, for example, pyridine, pyridazine, pyrimidine, pyrazine, triazine, pyran, pyrrole, pyrroline, imidazole, imidazoline, pyrazole, pyrazoline, triazole, tetrazole, thiophene, thiazole, isothiazole, thiadiazole, furan, oxazole, isoxazole, oxadiazole, oxathiadiazole, and the fused bicyclic moieties formed by fusing one of these monocyclic groups with a phenyl ring or with any of the heteroaromatic monocyclic groups to form a C8-C10 bicyclic group such as indole, benzimidazole, indazole, benzotriazole, isoquinoline, quinoline, benzothiazole, benzofur
• Aryl and heteroaryl moieties may be substituted with a variety of substituents including C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C5-C12 aryl, C1-C8 acyl, and heteroforms of these, each of which can itself be further substituted; other substituents for aryl and heteroaryl moieties include halo, OR, NR 2 , SR, SO 2 R, SO 2 NR 2 , NRSO 2 R, NRCONR 2 , NRCOOR, NRCOR, CN, C ⁇ CR, COOR, CONR 2 , OOCR, COR, and NO 2 , wherein each R is independently H, C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C6-C10 aryl, C
• an arylalkyl substituent may be substituted on the aryl portion with substituents described herein as typical for aryl groups, and it may be further substituted on the alkyl portion with substituents described herein as typical or suitable for alkyl groups.
• arylalkyl and “heteroarylalkyl” refer to aromatic and heteroaromatic ring systems which are bonded to their attachment point through a linking group such as an alkylene, including substituted or unsubstituted, saturated or unsaturated, cyclic or acyclic linkers.
• the linker is C1-C8 alkyl or a hetero form thereof.
• These linkers may also include a carbonyl group, thus making them able to provide substituents as an acyl or heteroacyl moiety.
• An aryl or heteroaryl ring in an arylalkyl or heteroarylalkyl group may be substituted with the same substituents described above for aryl groups.
• an arylalkyl group includes a phenyl ring optionally substituted with the groups defined above for aryl groups and a C1-C4 alkylene that is unsubstituted or is substituted with one or two C1-C4 alkyl groups or heteroalkyl groups, where the alkyl or heteroalkyl groups can optionally cyclize to form a ring such as cyclopropane, dioxolane, or oxacyclopentane.
• a heteroarylalkyl group preferably includes a C5-C6 monocyclic heteroaryl group that is optionally substituted with the groups described above as substituents typical on aryl groups and a C1-C4 alkylene that is unsubstituted or is substituted with one or two C1-C4 alkyl groups or heteroalkyl groups, or it includes an optionally substituted phenyl ring or C5-C6 monocyclic heteroaryl and a C1-C4 heteroalkylene that is unsubstituted or is substituted with one or two C1-C4 alkyl or heteroalkyl groups, where the alkyl or heteroalkyl groups can optionally cyclize to form a ring such as cyclopropane, dioxolane, or oxacyclopentane.
• substituents may be on either the alkyl or heteroalkyl portion or on the aryl or heteroaryl portion of the group.
• the substituents optionally present on the alkyl or heteroalkyl portion are the same as those described above for alkyl groups generally; the substituents optionally present on the aryl or heteroaryl portion are the same as those described above for aryl groups generally.
• Arylalkyl groups as used herein are hydrocarbyl groups if they are unsubstituted, and are described by the total number of carbon atoms in the ring and alkylene or similar linker.
• a benzyl group is a C7-arylalkyl group
• phenylethyl is a C8-arylalkyl.
• Heteroarylalkyl refers to a moiety comprising an aryl group that is attached through a linking group, and differs from “arylalkyl” in that at least one ring atom of the aryl moiety or one atom in the linking group is a heteroatom selected from N, O and S.
• the heteroarylalkyl groups are described herein according to the total number of atoms in the ring and linker combined, and they include aryl groups linked through a heteroalkyl linker; heteroaryl groups linked through a hydrocarbyl linker such as an alkylene; and heteroaryl groups linked through a heteroalkyl linker.
• C7-heteroarylalkyl would include pyridylmethyl, phenoxy, and N-pyrrolylmethoxy.
• Alkylene refers to a divalent hydrocarbyl group; because it is divalent, it can link two other groups together. Typically it refers to -(CHa) n - where n is 1-8 and preferably n is 1-4, though where specified, an alkylene can also be substituted by other groups, and can be of other lengths, and the open valences need not be at opposite ends of a chain. Thus -CH(Me)- and -C(Me )2- may also be referred to as alkylenes, as can a cyclic group such as cyclopropan- l,l-diyl.
• the substituents include those typically present on alkyl groups as described herein.
• any alkyl, alkenyl, alkynyl, acyl, or aryl or arylalkyl group or any heteroform of one of these groups that is contained in a substituent may itself optionally be substituted by additional substituents.
• the nature of these substituents is similar to those recited with regard to the primary substituents themselves if the substituents are not otherwise described.
• R 7 is alkyl
• this alkyl may optionally be substituted by the remaining substituents listed as embodiments for R 7 where this makes chemical sense, and where this does not undermine the size limit provided for the alkyl per se; e.g., alkyl substituted by alkyl or by alkenyl would simply extend the upper limit of carbon atoms for these embodiments, and is not included.
• each such alkyl, alkenyl, alkynyl, acyl, or aryl group may be substituted with a number of substituents according to its available valences; in particular, any of these groups may be substituted with fluorine atoms at any or all of its available valences, for example.
• Heteroform refers to a derivative of a group such as an alkyl, aryl, or acyl, wherein at least one carbon atom of the designated carbocyclic group has been replaced by a heteroatom selected from N, O and S.
• the heteroforms of alkyl, alkenyl, alkynyl, acyl, aryl, and arylalkyl are heteroalkyl, heteroalkenyl, heteroalkynyl, heteroacyl, heteroaryl, and heteroarylalkyl, respectively. It is understood that no more than two N, O or S atoms are ordinarily connected sequentially, except where an oxo group is attached to N or S to form a nitro or sulfonyl group.
• Halo as used herein includes fluoro, chloro, bromo and iodo. Fluoro and chloro are often preferred.
• Amino refers to NH 2 , but where an amino is described as “substituted” or “optionally substituted”, the term includes NR' R" wherein each R' and R" is independently H, or is an alkyl, alkenyl, alkynyl, acyl, aryl, or arylalkyl group or a heteroform of one of these groups, and each of the alkyl, alkenyl, alkynyl, acyl, aryl, or arylalkyl groups or heteroforms of one of these groups is optionally substituted with the substituents described herein as suitable for the corresponding group.
• R' and R" are linked together to form a 3-8 membered ring which may be saturated, unsaturated or aromatic and which contains 1-3 heteroatoms independently selected from N, O and S as ring members, and which is optionally substituted with the substituents described as suitable for alkyl groups or, if NR'R" is an aromatic group, it is optionally substituted with the substituents described as typical for heteroaryl groups.
• carbocyclic and heterocyclic structures encompass compounds having monocyclic, bicyclic or multiple ring systems, and may also comprise aryl or heteroaryl rings.
• heteroatom refers to any atom that is not carbon or hydrogen, such as nitrogen, oxygen or sulfur.
• heterocycles include but are not limited to pyridine, pyridazine, pyrimidine, pyrazine, triazine, pyran, pyrrole, pyrroline, imidazole, imidazoline, pyrazole, pyrazoline, triazole, tetrazole, thiophene, thiazole, isothiazole, thiadiazole, furan, oxazole, isoxazole, oxadiazole, oxathiadiazole, benzimidazole, benzofuran, isobenzofuran, indole, benzothiazole, benzotriazole, isoquinoline, quinoline, pyrazolopyridine, quinazoline, quinoxaline, cinnoline, tetrahydrofuran, 1,3-dioxolane, 2,3-dihydrofuran, tetrahydropyran, 1
• inorganic substituent refers to substituents that do not contain carbon or contain carbon bound to elements other than hydrogen (e.g., elemental carbon, carbon monoxide, carbon dioxide, and carbonate).
• inorganic substituents include but are not limited to nitro, halogen, azido, cyano, sulfonyls, sulfinyls, sulfonates, phosphates, etc.
• a candidate molecule or compound described herein may be in a therapeutically effective amount in a formulation or medicament, which is an amount that can lead to a biological effect, such as apoptosis of certain cells (e.g., cancer cells), reduction of proliferation of certain cells, or lead to ameliorating, alleviating, lessening, or removing symptoms of a disease or condition, for example.
• the terms also can refer to reducing or stopping a cell proliferation rate (e.g., slowing or halting tumor growth) or reducing the number of proliferating cancer cells (e.g., removing part or all of a tumor).
• microorganism examples include but are not limited to virus, bacterium and fungus.
• apoptosis refers to an intrinsic cell self-destruction or suicide program.
• cells undergo a cascade of events including cell shrinkage, blebbing of cell membranes and chromatic condensation and fragmentation. These events culminate in cell conversion to clusters of membrane-bound particles (apoptotic bodies), which are thereafter engulfed by macrophages.
• the invention in part provides pharmaceutical compositions comprising at least one compound within the scope of the invention as described herein, and methods of using compounds described herein.
• the invention in part provides methods for identifying a candidate molecule that interacts with a CK2 protein, which comprises contacting a composition containing a CK2 protein and a molecule described herein with a candidate molecule and determining whether the amount of the molecule described herein that interacts with the protein is modulated, whereby a candidate molecule that modulates the amount of the molecule described herein that interacts with the protein is identified as a candidate molecule that interacts with the protein.
• the invention provides methods for identifying a candidate molecule that interacts with a Pirn or Fit protein, which comprises contacting a composition containing a Pirn or Fit protein and a molecule described herein with a candidate molecule and determining whether the amount of the molecule described herein that interacts with the protein is modulated, whereby a candidate molecule that modulates the amount of the molecule described herein that interacts with the protein is identified as a candidate molecule that interacts with the protein.
• Also provided are methods for modulating the activity of a CK2 protein which comprises contacting a system comprising the protein with a compound described herein in an amount effective for modulating (e.g., inhibiting) the activity of the protein.
• the system in such embodiments can be a cell-free system or a system comprising cells.
• methods for reducing cell proliferation, and optionally inducing apoptosis which comprises contacting cells with a compound described herein in an amount effective to reduce proliferation of the cells.
• the cells in such embodiments can be in a cell line, in a tissue or in a subject (e.g., a research animal or human).
• compositions comprising a compound described herein in combination with a protein or cell, such as an isolated protein (e.g., isolated CK2 or other serine-threonine protein kinase protein) or a cell in a cell line (e.g., HCT-1 16 cell line).
• a protein or cell such as an isolated protein (e.g., isolated CK2 or other serine-threonine protein kinase protein) or a cell in a cell line (e.g., HCT-1 16 cell line).
• Serine-threonine protein kinases catalyze the transfer of a gamma phosphate from adenosine triphosphate to a serine or threonine amino acid in a peptide or protein substrate.
• methods which comprise contacting a system comprising a serine-threonine protein kinase protein with a compound described herein in an amount effective for modulating (e.g., inhibiting) the activity of the protein.
• the activity of the serine-threonine protein kinase is the catalytic activity of the protein (e.g., catalyzing the transfer of a gamma phosphate from adenosine triphosphate to a peptide or protein substrate).
• methods for identifying a candidate molecule that interacts with a serine-threonine protein kinase comprise: contacting a composition containing a serine- threonine protein kinase and a compound described herein with a candidate molecule under conditions in which the compound and the protein interact, and determining whether the amount of the compound that interacts with the protein is modulated relative to a control interaction between the compound and the protein without the candidate molecule, whereby a candidate molecule that modulates the amount of the compound interacting with the protein relative to the control interaction is identified as a candidate molecule that interacts with the protein.
• Systems in such embodiments can be a cell-free system or a system comprising cells (e.g., in vitro).
• the protein, the compound or the molecule in some embodiments is in association with a solid phase.
• the interaction between the compound and the protein is detected via a detectable label, where in some embodiments the protein comprises a detectable label and in certain embodiments the compound comprises a detectable label.
• the interaction between the compound and the protein sometimes is detected without a detectable label.
• Protein kinases catalyze the transfer of a gamma phosphate from adenosine triphosphate to a serine or threonine amino acid (serine/threonine protein kinase), tyrosine amino acid (tyrosine protein kinase), tyrosine, serine or threonine (dual specificity protein kinase) or histidine amino acid (histidine protein kinase) in a peptide or protein substrate.
• methods which comprise contacting a system comprising a protein kinase protein with a compound described herein in an amount effective for modulating (e.g., inhibiting) the activity of the protein kinase.
• the activity of the protein kinase is the catalytic activity of the protein (e.g., catalyzing the transfer of a gamma phosphate from adenosine triphosphate to a peptide or protein substrate).
• provided are methods for identifying a candidate molecule that interacts with a protein kinase which comprise: contacting a composition containing a protein kinase and a compound described herein with a candidate molecule under conditions in which the compound and the protein kinase interact, and determining whether the amount of the compound that interacts with the protein kinase is modulated relative to a control interaction between the compound and the protein kinase without the candidate molecule, whereby a candidate molecule that modulates the amount of the compound interacting with the protein kinase relative to the control interaction is identified as a candidate molecule that interacts with the protein kinase.
• Systems in such embodiments can be a cell-free system or a system comprising cells (e.g., in vitro).
• the protein kinase, the compound or the molecule in some embodiments is in association with a solid phase.
• the interaction between the compound and the protein kinase is detected via a detectable label, where in some embodiments the protein kinase comprises a detectable label and in certain embodiments the compound comprises a detectable label.
• the interaction between the compound and the protein kinase sometimes is detected without a detectable label.
• compositions of matter comprising a protein kinase and a compound described herein.
• the protein kinase in the composition is a serine-threonine protein kinase or a tyrosine protein kinase.
• the protein kinase is a protein kinase fragment having compound-binding activity.
• the protein kinase in the composition is, or contains a subunit (e.g., catalytic subunit, SH2 domain, SH3 domain) of, CK2, Pirn subfamily protein kinase (e.g., PDvIl, PIM2, PDvO) or Fit subfamily protein kinase (e.g, FLTl , FLT3, FLT4).
• the composition is cell free and sometimes the protein kinase is a recombinant protein.
• the protein kinase can be from any source, such as cells from a mammal, ape or human, for example.
• the protein kinase is a human protein kinase.
• the serine-threonine protein kinase can be from any source, such as a mammal, ape or human, for example.
• serine-threonine protein kinases that may be inhibited by compounds disclosed herein include without limitation human versions of CK2, CK2 ⁇ 2, Pirn subfamily kinases (e.g., PIMl, PIM2, PIM3), CDKl/cyclinB, c-RAF, Mer, MELK, HIPK3, HIPK2, ZIPK and ZIPK.
• a serine-threonine protein kinase sometimes is a member of a subfamily containing one or more of the following amino acids at positions corresponding to those listed in human CK2: leucine at position 45, methionine at position 163 and isoleucine at position 174.
• Examples of such protein kinases include without limitation human versions of CK2, STKlO, HIPK2, HIPK3, DAPK3, DYK2 and PIM- 1.
• Examples of tyrosine protein kinases that can be inhibited, or may potentially be inhibited, by compounds disclosed herein include without limitation human versions of Fit subfamily members (e.g., FLTl , FLT2, FLT3, FLT3 (D835Y), FLT4).
• DYRK2 An example of a dual specificity protein kinase that can be inhibited, or may potentially be inhibited, by compounds disclosed herein includes without limitation DYRK2. Nucleotide and amino acid sequences for serine-threonine protein kinases and reagents are publicly available (e.g., World Wide Web URLs ncbi.nlm.nih.gov/sites/entrez/ and Invitrogen.com).
• nucleotide sequences can be accessed using the following accession numbers: NM_002648.2 and NP_002639.1 for PIMl ; NM_006875.2 and NP_006866.2 for PIM2; XM_938171.2 and XP_943264.2 for PIM3; NM_0041 19.2 and NP_0041 10.2 for FLT3; NM_002020.3 and NP_00201 1.2 for FLT4; and NM_002019.3 and NP_002010.2 for FLTl.
• the invention also in part provides methods for treating a condition related to aberrant cell proliferation.
• methods of treating a cell proliferative condition in a subject which comprises administering a compound described herein to a subject in need thereof in an amount effective to treat the cell proliferative condition.
• the subject may be a research animal (e.g., rodent, dog, cat, monkey), optionally containing a tumor such as a xenograft tumor (e.g., human tumor), for example, or may be a human.
• a cell proliferative condition sometimes is a tumor or non-tumor cancer, including but not limited to, cancers of the colorectum, breast, lung, liver, pancreas, lymph node, colon, prostate, brain, head and neck, skin, liver, kidney, blood and heart (e.g., leukemia, lymphoma, carcinoma).
• cancers of the colorectum, breast, lung, liver, pancreas, lymph node, colon, prostate, brain, head and neck, skin, liver, kidney, blood and heart e.g., leukemia, lymphoma, carcinoma.
• methods of treating pain in a subject which comprise administering a compound described herein to a subject in need thereof in an amount effective to treat the pain.
• methods of treating inflammation in a subject which comprises administering a compound described herein to a subject in need thereof in an amount effective to treat the inflammation.
• the subject may be a research animal (e.g., rodent, dog, cat, monkey), for example, or may be a human.
• Conditions associated with inflammation and pain include without limitation acid reflux, heartburn, acne, allergies and sensitivities, Alzheimer's disease, asthma, atherosclerosis, bronchitis, carditis, celiac disease, chronic pain, Crohn's disease, cirrhosis, colitis, dementia, dermatitis, diabetes, dry eyes, edema, emphysema, eczema, fibromyalgia, gastroenteritis, gingivitis, heart disease, hepatitis, high blood pressure, insulin resistance, interstitial cystitis, joint pain/arthritis/rheumatoid arthritis, metabolic syndrome (syndrome X), myositis, nephritis, obesity, osteopenia, osteoporosis, Parkinson's disease, periodontal disease, polyarteritis, polychondritis, psoriasis, scleroderma, sinusitis, Sjogren's syndrome, spastic colon, systemic candidia
• a compound herein reduces inflammation or pain which comprise contacting a system with a compound described herein in an amount effective for modulating (e.g., inhibiting) the activity of a pain signal or inflammation signal.
• methods for identifying a compound that reduces inflammation or pain which comprise: contacting a system with a compound of Formula (I), (II), (III), (FV), (V) or (VI); and detecting a pain signal or inflammation signal, whereby a compound that modulates the pain signal relative to a control molecule is identified as a compound that reduces inflammation of pain.
• pain signals are formalin-stimulated pain behaviors and examples of inflammation signals include without limitation a level of a pro-inflammatory molecule.
• the invention also in part pertains to methods for modulating angiogenesis in a subject, and methods for treating a condition associated with aberrant angiogenesis in a subject.
• methods for determining whether a compound herein modulates angiogenesis which comprise contacting a system with a compound described herein in an amount effective for modulating (e.g., inhibiting) angiogenesis or a signal associated with angiogenesis.
• Signals associated with angiogenesis are levels of a pro-angiogenesis growth factor such as VEGF.
• Methods for assessing modulation of angiogenesis also are known, such as analyzing human endothelial tube formation (BD BioCoatTM Angiogenesis System from BD Biosciences).
• identifying a compound that modulates angiogenesis which comprise contacting a system with a compound of Formula (I), (II), (III), (IV), (V) or (VI) or a pharmaceutically acceptable salt thereof; and detecting angiogenesis in the system or an angiogenesis signal, whereby a compound that modulates the angiogenesis or angiogenesis signal relative to a control molecule is identified as a compound that modulates angiogenesis.
• methods for treating an angiogenesis condition which comprise administering a compound described herein to a subject in need thereof in an amount effective to treat the angiogenesis condition.
• Angiogenesis conditions include without limitation solid tumor cancers, varicose disease and the like.
• the invention also in part pertains to methods for modulating an immune response in a subject, and methods for treating a condition associated with an aberrant immune response in a subject.
• methods for determining whether a compound herein modulates an immune response which comprise contacting a system with a compound described herein in an amount effective for modulating (e.g., inhibiting) an immune response or a signal associated with an immune response.
• Signals associated with immunomodulatory activity include, e.g., stimulation of T-cell proliferation, suppression or induction of cytokines, including, e.g., interleukins, interferon- ⁇ and TNF.
• identifying a compound that modulates an immune response comprise contacting a system with a compound of Formula (I), (II), (III), (IV), (V) or (VI) or a pharmaceutically acceptable salt thereof; and detecting immunomodulatory activity in a system, or a signal associated with immunomodulatory activity, whereby a compound that modulates the immune response relative to a control molecule is identified as an immune response modulatory compound.
• methods for treating a condition associated with an aberrant immune response in a subject which comprise administering a compound described herein to a subject in need thereof in an amount effective to treat the condition.
• an immune response may be modulated by administering a compound herein in combination with a molecule that modulates (e.g., inhibits) the biological activity of an mTOR pathway member or member of a related pathway (e.g., mTOR, PI3 kinase, AKT).
• a molecule that modulates e.g., inhibits
• the biological activity of an mTOR pathway member or member of a related pathway e.g., mTOR, PI3 kinase, AKT.
• the molecule that modulates the biological activity of an mTOR pathway member or member of a related pathway is rapamycin.
• a composition comprising a compound described herein in combination with a molecule that modulates the biological activity of an mTOR pathway member or member of a related pathway, such as rapamycin, for example.
• the compound is a compound of Formula (I), (II), (III), (IV), (V) or (VI) in one of the Tables provided herein, or a pharmaceutically acceptable salt of one of these compounds.
• Any suitable formulation of a compound described above can be prepared for administration.
• Any suitable route of administration may be used, including, but not limited to, oral, parenteral, intravenous, intramuscular, transdermal, topical and subcutaneous routes.
• the mode of administration, and the type of treatment desired e.g., prevention, prophylaxis, therapy; the compounds are formulated in ways consonant with these parameters.
• Preparation of suitable formulations for each route of administration are known in the art. A summary of such formulation methods and techniques is found in Remington's Pharmaceutical Sciences, latest edition, Mack Publishing Co., Easton, PA, which is incorporated herein by reference.
• each substance or of the combination of two substances will generally include a diluent as well as, in some cases, adjuvants, buffers, preservatives and the like.
• the substances to be administered can be administered also in liposomal compositions or as microemulsions.
• formulations can be prepared in conventional forms as liquid solutions or suspensions or as solid forms suitable for solution or suspension in liquid prior to injection or as emulsions.
• Suitable excipients include, for example, water, saline, dextrose, glycerol and the like.
• Such compositions may also contain amounts of nontoxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents and the like, such as, for example, sodium acetate, sorbitan monolaurate, and so forth.
• Systemic administration may also include relatively noninvasive methods such as the use of suppositories, transdermal patches, transmucosal delivery and intranasal administration.
• Oral administration is also suitable for compounds of the invention. Suitable forms include syrups, capsules, tablets, as is understood in the art.
• the appropriate dosage of a compound described above often is 0.01-150 mg/kg, and sometimes 0.1-100 mg/kg. Dosage levels are dependent on the nature of the condition, drug efficacy, the condition of the patient, the judgment of the practitioner, and the frequency and mode of administration; however, optimization of such parameters is within the ordinary level of skill in the art.
• Compounds of the invention may be used alone or in combination with another therapeutic agent.
• the invention provides methods to treat conditions such as cancer and inflammation by administering to a subject in need of such treatment a therapeutically effective amount of a therapeutic agent that binds to certain DNA segments and administering to the same subject a CK2 modulator in an amount that is effective to enhance the activity of the therapeutic agent.
• a CK2 modulator is an agent that inhibits or enhances a biological activity of a CK2 protein, and is generically referred to hereafter as a "modulator.”
• a Pirn or Fit modulator is an agent that inhibits or enhances a biological activity of a Pirn protein or a FH protein.
• the therapeutic agent and the modulator may be administered together, either as separate pharmaceutical compositions or admixed in a single pharmaceutical composition.
• the therapeutic agent and the modulator may also be administered separately, including at different times and with different frequencies, as long as the modulator is administered at a time that increases the potency of the therapeutic agent.
• the modulator may be administered by any known route, such as orally, intravenously, intramuscularly, nasally, and the like; and the therapeutic agent may also be administered by any conventional route. In many embodiments, at least one and optionally both of the modulator and the therapeutic agent may be administered orally.
• the modulator and the therapeutic agent are administered at the same time, whether in separate dosages or admixed in a single dosage.
• the modulator and therapeutic agent are preferably combined into a single pharmaceutical composition, so the treated patient may receive a single oral dosage or a single injection, for example.
• the amount of each of these materials to be administered will vary with the route of administration, the condition of the subject, other treatments being administered to the subject, and other parameters.
• the therapeutic agents of the invention may, of course, cause multiple desired effects; and the amount of modulator to be used in combination with the therapeutic agent should be an amount that increases one or more of these desired effects.
• the modulator is to be administered in an amount that is effective to enhance a desired effect of the therapeutic agent.
• An amount is "effective to enhance a desired effect of the therapeutic agent", as used herein, if it increases by at least about 25% at least one of the desired effects of the therapeutic agent alone.
• it is an amount that increases a desired effect of the therapeutic agent by at least 50% or by at least 100% (i.e., it doubles the effective activity of the therapeutic agent.) In some embodiments, it is an amount that increases a desired effect of the therapeutic agent by at least 200%.
• the amount of a modulator that increases a desired effect of a therapeutic agent may be determined using in vitro methods, such as cell proliferation assays.
• the therapeutic agents of the invention are useful to counter hyperproliferative disorders such as cancer, thus they reduce cell proliferation.
• a suitable amount of a modulator could be the amount needed to enhance an antiproliferative effect of a therapeutic agent by at least 25% as determined in a cell proliferation assay.
• the modulator used in the present invention enhances at least one desired effect produced by the therapeutic agent it is used with, thus the combinations of the inveniton provide a synergistic effect, not merely an additive effect.
• the modulators themselves are at times useful for treating the same types of conditons, and thus may also have some direct effect in such assays. In that event, the "amount effective to increase a desired effect" must be a synergistic enhancement of the activity of the therapeutic agent that is attributable to enhancement by the modulator of an effect of the therapeutic agent, rather than a simple additive effect that would be expected with separate administration of the two materials.
• the modulator can be used in an amount (concentration) that would not be expected to have any apparent effect on the treated subject or the in vitro assay, so the increased effect achieved with the combination is directly attributable to a synergistic effect.
• the present invention includes methods and compositions for treating a patient having a cell proliferation disorder or an inflammatory disorder with a therapeutic agent as described herein, and a "modulator" described above, where the timing of administration of the modulator permits it to enhance a desired effect of the therapeutic agent .
• a "modulator" as described above may be used in combination with a therapeutic agent that can act by binding to regions of DNA that can form certain quadruplex structures.
• the therapeutic agents have anticancer activity on their own, but their activity is enhanced when they are used in combination with a modulator. This synergistic effect allows the therapeutic agent to be administered in a lower dosage while achieving equivalent or higher levels of at least one desired effect.
• the appropriate dosage of a modulator such as a compound of Formula (I), (II), (III), (IV), (V), or (VI) as described herein, is typically between about 0.01-15 mg/kg, and about 0.1-10 mg/kg. Dosage levels are dependent on the nature of the condition, drug efficacy, the condition of the patient, the judgment of the practitioner, and the frequency and mode of administration; however, optimization of such parameters is within the ordinary level of skill in the art.
• a modulator may be separately active for treating a cancer.
• the dosage of a modulator will frequently be two-fold to ten-fold lower than the dosage required when the modulator is used alone to treat the same condition or subject. Determination of a suitable amount of the modulator for use in combination with a therapeutic agent is readily determined by methods known in the art.
• the therapeutic agents of the invention are compounds that bind to certain motifs in nucleic acids.
• the therapeutic agent to be used can be selected from several different classes of compounds, such as those that bind to quadruplex-forming regions of DNA.
• the therapeutic agents are useful for the treatment of cancer and other indications such as inflammatory disorders, and methods for making and using them are known in the art. Several preferred classes of these therapeutic agents are described below. Each class of therapeutic agents can be used in combination with any active CK modulator, including but not limited to those disclosed herein.
• the therapeutic agent can be a compound of formula (TAl-I):
• V is H, halo, or NR 1 R 2 ;
• A is H, fluoro, or NR 1 ⁇ ;
• Z is O, S, NR 1 or CH 2 ;
• X is OR 2 , NR 1 R 2 , halo, azido, or SR 2 ; n is 1-3; wherein in NR 1 R 2 , R 1 and R 2 may form a double bond or a ring, each of which is optionally substituted;
• R 1 is H or a Ci. 6 alkyl
• W is an optionally substituted aryl or heteroaryl, which may be monocyclic or fused with a single or multiple ring and optionally containing a heteroatom; or a compound having formula (TA 1-2):
• V, A, X, Z and U are as defined in formula TAl-I, and W is selected from the group consisting of
• the therapeutic agent is a compound having formula (TAl- IA): or a pharmaceutically acceptable salt, esters or prodrug thereof, or a specific isomer or mixture of isomers thereof.
• the therapeutic agent of the combinations of the invention is a compound of this formula:
• V is H, halo, or NR 1 R 2 ; AisH, fluoro, orNR' 2 ; Z is O, S, NR 1 OrCH 2 ; UiSOR 2 OrNR 1 R 2 ; X is OR 2 , NR 1 R 2 , halo, azido, or SR 2 ; n is 1-3; wherein in NR 1 R 2 , R 1 and R 2 may form a double bond or a ring, each of which is optionally substituted;
• R 1 isHoraC,. 6 alkyl
• R is H or a C
• R 2 is an optionally substituted heterocyclic ring, aryl or heteroaryl
• W is an optionally substituted aryl or heteroaryl, which may be monocyclic or fused with a single or multiple ring and optionally containing a heteroatom; or a compound having formula (TA3-2)
• V, A, X, Z and U are as defined in formula 1 , and W is selected from the group consisting of
• the therapeutic agent of the combinations of the invention is a compound of this formula:
• B, X, A, or V is absent if Z 2 , Z 3 , or Z 4 , respectively, is N , and independently H, halo, azido, R 2 , CH 2 R 2 , SR 2 , OR 2 or NR 1 R 2 if Z 2 , Z 3 , or Z 4 , respectively, is C; or
• a and V, A and X, or X and B may form a carbocyclic ring, heterocyclic ring, aryl or heteroaryl, each of which may be optionally substituted and/or fused with a cyclic ring;
• Z 1 , Z 2 , Z 3 and Z 4 are C or N, provided any two N are non-adjacent;
• W together with N and Z forms an optionally substituted 5- or 6-membered ring that is fused to an optionally substituted saturated or unsaturated ring;
• said saturated or unsaturated ring may contain a heteroatom and is monocyclic or fused with a single or multiple carbocyclic or heterocyclic rings;
• R 4 is H, a C] - 1O alkyl or C 2 .io alkenyl optionally containing one or more non-adjacent heteroatoms selected from N, O and S, and optionally substituted with a carbocyclic or heterocyclic ring; or R 3 and R 4 together with N may form an optionally substituted ring; • each R 5 is a substituent at any position on ring W; and is H, OR 2 , amino, alkoxy, amido, halogen, cyano or an inorganic substituent; or R 5 is C ⁇ _6 alkyl, C 2 .
• B may be absent when Z 1 is N, or is H or a halogen when Z 1 is C.
• W together with N and Z form a group having the formula selected from the group consisting of
• R 6 is H, or a substituent known in the art, including but not limited to hydroxyl, alkyl, alkoxy, halo, amino, or amido; and ring S and ring T may be saturated or unsaturated.
• W together with N and Z forms a 5- or 6-membered ring that is fused to a phenyl. In other embodiments, W together with N and Z forms a 5- or 6-membered ring that is optionally fused to another ring, when U is NR 1 R 2 , provided U is not NH 2 . In certain embodiments, W together with N and Z forms a 5- or 6-membered ring that is not fused to another ring, when U is NR 1 R 2 (e.g., NH 2 ).
• the compounds of the present invention have the general formula (TA4-2A) or (TA4-2B):
• A, B, V, X, U, Z, Z 1 , Z 2 , Z 3 , Z 4 and n are as described for TA4-1 ;
• R 6 is H, Ci- 6 alkyl, hydroxyl, alkoxy, halo, amino or amido;
• Z and Z 5 may optionally form a double bond.
• U may be NR l 1 rR> 2', wherein R 1 is H, and R 2 is a C M0 alkyl optionally substituted with a heteroatom, a C3.6 cycloalkyl, aryl or a 5-14 membered heterocyclic ring containing one or more N, O or S.
• R 2 may be a Ci -I0 alkyl substituted with an optionally substituted morpholine, thiomorpholine, imidazole, aminodithiadazole, pyrrolidine, piperazine, pyridine or piperidine.
• R 1 and R 2 together with N form an optionally substituted piperidine, pyrrolidine, piperazine, morpholine, thiomorpholine, imidazole, or aminodithiazole.
• the therapeutic agent to be combined with a CK modulator can be selected from compounds having this formula:
• V, X, and Y are absent if attached to a heteroatom other than Nitrogen, and independently H, halo, azido, R 2 , CH 2 R 2 , SR 2 , OR 2 or NR 1 R 2 when attached to C or N; or wherein V and X, or X and Y may form a carbocyclic ring, heterocyclic ring, aryl or hdteroaryl, each of which may be optionally substituted and/or fused with a cyclic ring;
• Z 1 , Z 2 and Z 3 are C, N, O or S;
• W together with N and Z forms an optionally substituted 5- or 6-membered ring that is fused to an optionally substituted aryl or heteroaryl, wherein said aryl or heteroaryl may be monocyclic or fused with a single or multiple ring, and wherein said ring optionally contains a heteroatom;
• R 1 and R 3 are independently H or C
• R 4 is H, a Ct-io alkyl or C 2 . ⁇ o alkenyl optionally containing one or more non-adjacent heteroatoms selected from N, O and S, and optionally substituted with a carbocyclic or heterocyclic ring; or R 3 and R 4 together with N may form an optionally substituted ring; each R 5 is a substituent at any position on W; and is H, OR 2 , amino, alkoxy, amido, halogen, cyano or an inorganic substituent; or R 5 is C ⁇ _ ⁇ alkyl, C 2 - 6 alkenyl, -CONHR 1 , each optionally substituted by halo, carbonyl or one or more non-adjacent heteroatoms; or two adjacent R 5 are linked to obtain a 5-6 membered optionally substituted carbocyclic or heterocyclic ring, optionally fused to an additional optionally substituted carbocyclic or heterocyclic ring; and n is 1-6.
• ring T may form an optionally substituted 5- membered ring selected from the group consisting of:
• W together with N and Z may form an optionally substituted 5- or 6-membered aryl or heteroaryl ring that is fused to an optionally substituted aryl or heteroaryl selected from the group consisting of:
• W together with N and Z may form a group having the formula selected from the group consisting of
• R 6 is H, or a substituent known in the art, including but not limited to hydroxyl, alky], alkoxy, halo, amino, or amido;
• Ring S and M may be saturated or unsaturated.
• W together with N and Z may form a 5- or 6-membered ring that is fused to a phenyl.
• the compounds of the present invention have the general formula (TA5-2A) or (TA5-2B): wherein U, V, W, X, Y, Z, Z ⁇ Z 2 , Z 3 , R 5 and n are as described above for TA5-1 ;
• Z and Z 4 may optionally form a double bond.
• U may be SO 2 NR 1 R 2 , wherein R 1 is H, and R 2 is a C
• R 2 may be a C]. 10 alkyl substituted with an optionally substituted morpholine, thiomo ⁇ holine, imidazole, aminodithiadazole, pyrrolidine, piperazine, pyridine or piperidine.
• R 1 and R 2 together with N form an optionally substituted piperidine, pyrrolidine, piperazine, morpholine, thiomo ⁇ holine, imidazole, or aminodithiazole.
• U is SO 2 NR'-(CR'2)n-NR 3 R 4 ; n is 1-4; each R 1 is H or alkyl; and R 3 and R 4 in NR 3 R 4 together form an optionally substituted piperidine, pyrrolidine, piperazine, mo ⁇ holine, thiomo ⁇ holine, imidazole, or aminodithiazole.
• U is SO 2 NH-(CH 2 ) n -NR 3 R 4 wherein R 3 and R 4 together with N form an optionally substituted pyrrolidine, which may be linked to (CH 2 ) n at any position in the pyrrolidine ring. In one embodiment, R 3 and R 4 together with N form an N-methyl substituted pyrrolidine.
• the present invention provides compounds having formula (TA5-1), (TA5-2A) or (TA5-2B), wherein: each of V and Y if present is independently H or halogen (e.g., chloro or fluoro);
• X is -(ROR 1 R', wherein R 3 is C or N and wherein in each -(ROR 1 1 DR2" , R 1 and R' together may form an optionally substituted aryl or heteroaryl ring;
• Z is NH or N-alkyl (e.g., N-CH 3 ); W together with N and Z forms an optionally substituted 5- or 6-membered ring that is fused with an optionally substituted aryl or heteroaryl ring; and
• U is -SO 2 R 5 R 6 -(CH 2 ) n -CHR 2 -NR 3 R 4 , wherein R 5 is CR 1 or N; R 1 is H or alkyl; R 6 is H or C
• the present invention provides compounds having formula (TA5- 1), (TA5-2A) or (TA5-2B), wherein:
• V and Y if present is H or halogen (e.g., chloro or fluoro);
• X if present is -(R 5 )R : R 2 , wherein R 5 is C or N and wherein in each -(R 5 )R ! R 2 , R 1 and R 2 together may form an optionally substituted aryl or heteroaryl ring;
• Z is NH or N-alkyl (e.g., N-CH 3 );
• U is -SO 2 R 5 R 6 -(CH 2 ) n -CHR 2 -NR 3 R 4 ,
• R 5 is CR 1 or N
• R 6 is H or alkyl and wherein in the -CHR 2 -NR 3 R 4 moiety each R 3 or R 4 together with the C may form an optionally substituted heterocyclic or heteroaryl ring, or wherein in the -CHR 2 - NR 3 R 4 moiety each R 3 or R 4 together with the N may form an optionally substituted carbocyclic, heterocyclic, aryl or heteroaryl ring.
• the compounds of the present invention have the general formula (TA5-3):
• the compounds of the present invention have the general formula (TA5-4A) or (TA5-4B):
• the therapeutic agent for the combinations of the invention can be a compound of the formula:
• X is H, OR", NR 1 R', halogen, azido, SR' or CH 2 R;
• A is H, halogen, NR 1 R', SR', OR", CH 2 R", azido or NR 1 - (CR 2 ) n - NR J R 4
• Z is O, S, NR 1 or CH 2 ;
• U is R", OR", NR > 1 1 ⁇ R> 2' or NR 1 - (CR' 2 ) n - NR 3 J ⁇ R> 4 4 . provided U is not H;
• W is an optionally substituted aryl or heteroaryl, which may be monocyclic or fused with a single or multiple ring optionally containing a heteroatom; wherein R 1 and R 2 together with N in NR 1 R 2 , and R 3 and R 4 together with N in NR 3 R 4 may independently form an optionally substituted 5-6 membered ring containing N, and optionally O or S;
• R 1 and R 3 are independently H or a C
• R 2 and R 4 are independently H, or a C MO alkyl or C 2 - 1 0 alkenyl optionally containing one or more non-adjacent heteroatoms selected from N, O, and S, and optionally substituted with a substituted or unsubstituted aryl, heteroaryl, carbocyclic, or heterocyclic ring; or R 2 is an optionally cycloalkyl, substituted heterocyclic ring, aryl or heteroaryl;
• W may be selected from the group consisting of
• Q, Q 1 , Q 2 , and Q 3 are independently CH or N;
• R 5 is as defined in formula 1.
• each W in the above formula TA6- 1 or TA6-1A may be an optionally substituted phenyl, pyridine, biphenyl, naphthalene, phenanthrene, quinoline, isoquinoline, quinazoline, cinnoline, phthalazine, quinoxaline, indole, benzimidazole, benzoxazole, benzthiazole, benzofuran, anthrone, xanthone, acridone, fluorenone, carbazolyl, pyrimido[4,3-b]furan, pyrido[4,3-b]indole, pyrido[2,3-6]indole, dibenzofuran, acridine or acridizine.
• W is an optionally substituted phenyl.
• the present invention utilizes the above therapeutic agents in combination with at least one modulator.
• Modulators that can be utilized in combination with a therapeutic agent described above include compounds having structures of Formula (I), (II), (III), (IV), (V) and (VI) as described herein.
• the present invention utilizes the above therapeutic agents in combination with at least one modulator.
• compounds of the invention may be used in combination with PARP inhibitors.
• PARP inhibitors are known in the art, and are disclosed, for example, in CR. Calebrese, et al., Clin. Cancer Res. vol. 9, 271 1-18 (2003); S.J. Veuger, et al., Cancer Res. vol. 63, 6008-15 (2003); CR. Calabrese et al., J. Nat 'I. Cancer Inst. 96( 1 ), 56-67 (2004); "Potent Novel PARP Inhibitors," Expert Reviews in Molecular Medicine, vol. 7(4) (March 2005); and P.
• PARP inhibitors disclosed in these documents are suitable for use in the methods and compositions of the present invention.
• Additional PARP inhibitors that can be used include, for example, 10-(4- methyl-piperazin-l-ylmethyl)-2H-7-oxa-l ,2-diaza-benzo[de]anthracen-3-one (GPI 15427) and 2-(4-methyl-piperazin-l-yl)-5H-benzo[c][ l,5]naphthyridin-6-one (GPI 16539). See Di Paola, et al., Eur. J. Pharmacology, 527(1-3), 163-71 (2005).
• Representative, but non-limiting, examples of PARP inhibitors that are suitable for use in the invention include the known compounds shown hereafter, including the pharmaceutically acceptable salts thereof, and individual isomers or mixtures of isomers thereof.
• Modulators that can be utilized in combination with a therapeutic agent described above also include compounds having structures of Formula (I), (II), (III), (IV), (V) and (VI) as described herein.
• the compound TAl-IA is a preferred therapeutic agent for use in the methods and compositions of the invention. More detail on suitable methods for its formulation and administration are provided in U.S. Provisional Application Serial No. 60/803,864 to Lim, et al., which was filed on June 3, 2006.
• compositions comprising at least one therapeutic agent within the scope of the invention as described herein in combination with at least one modulator.
• the composition may comprise a diluent or other pharmaceutically acceptable excipients.
• the appropriate dosage of the therapeutic agent is typically 0.01-15 mg/kg, preferably 0.1- 10 mg/kg. Dosage levels are dependent on the nature of the condition, drug efficacy, the condition of the patient, the judgment of the practitioner, and the frequency and mode of administration; however, optimization of such parameters is within the ordinary level of skill in the art.
• the dosage of a modulator is typically between about 0.01-150 mg/kg, and about 0.1-100 mg/kg.
• a modulator may be separately active for treating a cancer.
• the dosage of a modulator will frequently be two-fold to ten-fold lower than the dosage required when the modulator is used alone to treat the same condition or subject. Determination of a suitable amount of the modulator for use in combination with a therapeutic agent is readily determined by methods known in the art.
• Also provided are methods for modulating the activity of a CK protein which comprises contacting a system comprising the CK protein with a composition described herein in an amount effective for modulating (e.g., inhibiting) the activity of the protein.
• the system in such embodiments can be a cell-free system or a system comprising cells.
• methods for reducing cell proliferation, and optionally inducing apoptosis which comprises contacting cells with a composition or a combination therapy as described herein, wherein a therapeutic agent is administered in an amount effective to reduce proliferation of the cells, and a CK inhibitor is administered in an amount sufficient to enhance the efficacy of the therapeutic agent.
• the cells in such embodiments can be in a cell line, in a tissue or in a subject (e.g., a research animal or human).
• the invention also in part provides methods for treating a condition related to aberrant cell proliferation.
• methods of treating a cell proliferative condition in a subject which comprises administering a therapeutic agent described herein and a CK inhibitor described herein to a subject in need of treatment for a cell proliferative disorder; the therapeutic agent and the CK inhibitor are administered in amounts effective to treat the cell proliferative condition.
• the subject may be a research animal (e.g., rodent, dog, cat, monkey), optionally containing a tumor such as a xenograft tumor (e.g., human tumor), for example, or may be a human.
• a cell proliferative condition sometimes is a tumor or non-tumor cancer, including but not limited to, cancers of the colorectum, breast, lung, liver, pancreas, lymph node, colon, prostate, brain, head and neck, skin, liver, kidney, blood and heart (e.g., leukemia, lymphoma, carcinoma).
• cancers of the colorectum, breast, lung, liver, pancreas, lymph node, colon, prostate, brain, head and neck, skin, liver, kidney, blood and heart e.g., leukemia, lymphoma, carcinoma
• any suitable formulation of the therapeutic agent and the CK inhibitor can be prepared for administration, either together or separately. Any suitable route of administration may be used for each component, including but not limited to oral, parenteral, intravenous, intramuscular, transdermal, topical and subcutaneous routes.
• the two substances used together may be administered separately or together. When administered together, they may be in separate dosage forms, or they may be combined into a single combination drug.
• pharmaceutical compositions comprising a therapeutic agent as described herein and at least one CK inhibitor, and a pharmaceutically acceptable excipient.
• Ethyl 3-bromo-4-pyridine carboxylate 1.15 g, 5.0 mmol
• 2-amino-4- methoxycarbonyl-phenylboronic acid (1.04 g, 4.5 mmol)
• sodium acetate (1.64 g, 20 mmol)
• l ,r-bis(diphenylphosphino)ferrocene palladium (II) chloride complexed with dichloromethane
• dimethylformamide 7.5 mL
• Methyl 2-acetamido-4-oxo-4,5-dihydrothiazolo[4,5-c]quinoline-7-carboxylate was prepared according to the procedure used in process 2, starting from methyl 2-acetamido-5- bromothiazole-4-carboxylate. Methyl 2-acetamido-4-oxo-4,5-dihydrothiazolo[4,5-c]quinoline- 7-carboxylate was isolated as a black solid (106 mg, 37% yield). LCMS (ES): 95% pure, m/z 318 [M+1] + . Process 28
• Compounds of Formula (II) are prepared by routes analogous to those described herein for preparation of compounds of Formula (I).
• the halogenated products prepared in processes 20, 21 , 22, and 30, may be converted to compounds of Formula (II) under conditions substantially as described in Example 4, or under conditions known to those of skill in the art.
• Test compounds in aqueous solution are added at a volume of 10 microliters, to a reaction mixture comprising 10 microliters Assay Dilution Buffer (ADB; 2OmM MOPS, pH 7.2, 25 mM beta-glycerolphosphate, 5 mM EGTA, 1 mM sodium orthovanadate and 1 mM dithiothreitol), 10 microliters of substrate peptide (RRRDDDSDDD, dissolved in ADB at a concentration of 1 mM), 10 microliters of recombinant human CK2 (25 ng dissolved in ADB; Upstate).
• ADB Assay Dilution Buffer
• RRRDDDSDDD substrate peptide
• Reactions are initiated by the addition of 10 microliters of ATP Solution (90% 75 mM MgCl 2 , 75 micromolar ATP dissolved in ADB; 10% [ ⁇ - 33 P]ATP (stock 1 mCi/100 ⁇ l; 3000 Ci/mmol (Perkin Elmer) and maintained for 10 minutes at 30 degrees C.
• the reactions are quenched with 100 microliters of 0.75% phosphoric acid, then transferred to and filtered through a phosphocellulose filter plate (Millipore). After washing each well 5 times with 0.75% phosphoric acid, the plate is dried under vacuum for 5 min and, following the addition of 15 ul of scintillation fluid to each well, the residual radioactivity is measured using a Wallac luminescence counter.
• a representative cell-proliferation assay protocol using Alamar Blue dye (stored at 4°C, use 20ul per well) is described hereafter.
• 96-well plate setup and compound treatment a. Split and trypsinize cells.
• H EMPTY d Add 100 ⁇ l of 2X drug dilution to each well in a concentration shown in the plate layout above. At the same time, add 100 ⁇ l of media into the control wells (wells B lO to B 12). Total volume is 200 ⁇ l /well.
• e Incubate four (4) days at 37°C, 5% CO2 in a humidified incubator.
• f. Add 20 ⁇ l Alamar Blue reagent to each well.
• g. Incubate for four (4) hours at 37°C, 5% CO2 in a humidified incubator.
• h Record fluorescence at an excitation wavelength of 544nm and emission wavelength of 590nm using a microplate reader.
• cells are cultured with a test compound for approximately four days, the dye is then added to the cells and fluorescence of non-reduced dye is detected after approximately four hours.
• Different types of cells can be utilized in the assays (e.g., HCT-1 16 human colorectal carcinoma cells, PC-3 human prostatic cancer cells and MiaPaca human pancreatic carcinoma cells).
• the human leukemia Jurkat T-cell line is maintained in RPMI 1640 (Cambrex) supplemented with 10% fetal calf serum and 50 ng/ml Geutamycin. Before treatment cells are washed, resuspended at a density of about 10 6 cells/milliliter in medium containing 1 % fetal calf serum and incubated in the presence of indicated mounts of drug for two hours.
• Cells are recovered by centrifugation, lysed using a hypotonic buffer (20 mM Tris/HCl pH 7.4; 2 mM EDTA; 5 mM EGTA; 10 mM mercaptoethanol; 1OmM NaF; 1 uM Okadaic acid; 10% v/v glycerol; 0.05% NP-40; 1% Protease Inhibitor Cocktail) and protein from the cleared lysate is diluted to 1 microgram per microliter in Assay Dilution Buffer (ADB; 2OmM MOPS, pH 7.2, 25mM ⁇ -glycerolphosphate, 5mM EGTA, ImM sodium orthovanadate and ImM dithiothreitol).
• ADB Assay Dilution Buffer
• the pharmacokinetics properties of drugs are investigated in ICR mice following an intravenous (FV) bolus and oral (PO) doses of drug at 5 mg/kg and 25 mg/kg respectively. Blood samples are collected at predetermined times and the plasma separated. Plasma is separated from the blood samples collected at 5, 15 and 30 minutes and 1 , 2, 4, 8 and 24 hours post-dose.
• FV intravenous
• PO oral
• Drug levels are quantified by the LC/MS/MS method described below.
• Noncompartmental pharmacokinetic analysis is applied for intravenous administration.
• a linear trapezoidal rule is used to compute AUC(O- 24).
• /2 and Co are calculated using the last three and the first three data points, respectively
• Bioanalysis is performed using a Quattro Micro LC/MS/MS instrument in the MRM detection mode, with an internal standard (IS). Briefly, 15 ⁇ L plasma samples are prepared for analysis using protein precipitation with 120 ⁇ L of acetonitrile. The supernatants are transferred into a 96 well plate and subjected to LC-MS/MS analysis using a Phenomenex Polar-RP HPLC column. The mobile phases are 10 mM NH 4 HCO 3 in water (Solution-A) and 10 mM NH 4 HCO 3 in methanol (Solution-B). The column is initially equilibrated with 25 % Solution-B and followed with 100% Solution B over 5 minutes. The method has a dynamic range from 1 to 10,000 ng/mL. Quantitation of the analytes is performed in the batch mode with two bracketing calibration curves according to the bioanalytical sample list.
• Compounds described herein are profiled for in vitro modulatory activity against protein kinases other than CK2.
• the in vitro analysis is conducted using known protocols (e.g., assay protocols described at world-wide web address upstate.com/img/pdf/KP_Assay Protocol_BookIet_v3.pdf).
• Compounds described herein are screened in the assays and prioritized based upon modulatory activity against protein kinases other than CK2 and specificity for CK2.
• test compounds in aqueous solution are added at a volume of 5 ⁇ l, to a reaction mixture comprising 5 ⁇ l of 5x Reaction buffer (4OmM MOPS, pH 7.0, ImM EDTA), 2.5 ⁇ l of recombinant human PDvI-I solution (10 ng), 2.5 ⁇ l of substrate peptide (KKRNRTLTK) and 10 ⁇ l of ATP solution - 98% (75 mM MgC12 37.5 uM ATP) 2% ([ ⁇ - 33P]ATP: 3000Ci/mmol - Perkin Elmer).
• 5x Reaction buffer 4OmM MOPS, pH 7.0, ImM EDTA
• 2.5 ⁇ l of recombinant human PDvI-I solution 10 ng
• substrate peptide KKRNRTLTK
• ATP solution - 98% 75 mM MgC12 37.5 uM ATP
• the reactions are incubated for 10 min at 30 0 C, quenched with 100 ⁇ l of 0.75% Phosphoric acid, then transferred to and filtered through a Phosphocellulose filter plate (Millipore). After washing each well 5 times with 0.75% Phosphoric acid, Scintillation fluid (15 ⁇ l) is added to each well. The residual radioactivity is measured using a luminescence counter.
• Test compounds are tested further for activity against other protein kinases.
• Selected kinase inhibition ICsodata are determined using standardized radiometric kinase assays for each individual kinase, which entail filter binding of 33 P labeled substrate proteins by the kinase of interest. Each IC 50 value is determined over a range of 10 drug concentrations. Representative reaction conditions are available from the World Wide Web URL upstate.corn/discovery/services/ic50_profiler.q.
• each Z 1 , Z 2 , Z 3 , and Z 4 is N, CH, or CR 3 , provided that none, one or two of Z 1 , Z 2 , Z 3 , and Z 4 are N; and further provided that one or more of Z 1 , Z 2 , Z 3 , and Z 4 is CR 3 ; each R 3 is independently an optionally substituted C1-C8 alkyl, C2-C8 heteroalkyl, C2-
• each R 3 is independently halo, CF 3 , CFN, OR, NR 2 , NROR, NRNR 2 , SR, SOR, SO 2 R,
• C2-C8 heteroalkyl C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl,
• each of R 6A , R 6B and R 6C can be halo, CF 3 , OR, NR 2 , NROR, NRNR 2 , SR, SOR, SO 2 R, SO 2 NR 2 , NRSO 2 R, NRCONR 2 , NRCOOR, NRCOR, CN, COOR, CONR 2 , 0OCR, COR, or NO 2 ; wherein each R is independently H or C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6- ClO
• R 10 is an optionally substituted five-membered or six-membered carbocyclic or heterocyclic aromatic ring.
• each of each Z 1 , Z 3 and Z 4 iiss CCHH,, aanndd ZZ 22 iiss CCRR 33 ,, wwhhcere R 3 is a carboxylic acid, carboxylate salt, ester, carboxamide, tetrazole or carboxy bioisostere.
• A6 The compound of any one of embodiments A1-A4, wherein at least one of Z'-Z 4 is N.
• A7 The compound of any one of embodiments A1-A6, wherein R 10 is an optionally substituted six-membered carbocyclic or heterocyclic aromatic ring.
• A12 The compound of any one of embodiments Al-Al 1, wherein at least one of R 6B or R 6C is H.
• A13 The compound of any one of embodiments A1-A12, wherein R 6B and R 6C are H.
• a 14 The compound of any one of embodiments Al -A 12, wherein R 6B is H, SR, SO 2 R or NR 2 , where each R is independently H or C M alkyl.
• a 15 The compound of of any one of embodiments A3-A14, wherein R 6 ⁇ is H.
• R 3 is a carboxylic acid, carboxylate salt, ester, carboxamide, tetrazole or carboxy bioisostere.
• B5. The compound of any one of embodiments B1-B4, wherein R 1 10 i,s an optionally substituted six-membered carbocyclic or heterocyclic aromatic ring.
• B6. The compound of embodiment B5, wherein R 10 is an optionally substituted phenyl or pyridyl ring.
• R 6C is H.
• B 12 The compound of any one of embodiments B l-B lO, wherein R 6B is H, SR, SO 2 R or NR?, where each R is independently H or C M alkyl.
• each Z 1 , Z 2 , Z 3 , and Z 4 is N, CH, or CR 3 , provided that none, one or two of Z 1 , Z 2 , Z 3 , and Z 4 are N; and further provided that one or more of Z 1 , Z 2 , Z 3 , and Z 4 is CR 3 ;
• each R 3 is independently an optionally substituted C1-C8 alkyl, C2-C8 heteroalkyl, C2- C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C 1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C12 heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkyl group, or each R 3 is independently halo, CF 3 , OR, NR 2 , NfR
• Z 5 is CR 6A or N; each of R 6A and R 6B is independently H or an optionally substituted C1 -C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C12 heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkyl group, or each of R 6A and R 6B can be halo, CF 3 , OR, NR 2 , NROR, NRNR 2 , SR, SOR, SO 2 R, SO 2 NR 2 , NRSO 2 R, NRCONR 2 , NRCOOR, NRCOR, CN, COOR, CONR 2 , OOCR, COR, or NO 2 ; wherein each R is independently
• R 10 is an optionally substituted five-membered or six-membered carbocyclic or heterocyclic aromatic ring.
• each of each Z 1 , Z 3 and Z 4 is CH, and Z 2 is CR 3 , where R 3 is a carboxylic acid, carboxylate salt, ester, carboxamide, tetrazole or carboxy bioisostere.
• a pharmaceutical composition comprising a compound of any one of embodiments Al -A 15 and a pharmaceutically acceptable excipient.
• a pharmaceutical composition comprising a compound of any one of embodiments Bl -B 13 and a pharmaceutically acceptable excipient.
• a pharmaceutical composition comprising a compound of any one of embodiments Cl -C 13 and a pharmaceutically acceptable excipient.
• a method for identifying a candidate molecule that interacts with a casein kinase 2 (CK2) protein comprises: contacting a composition containing a CK2 protein and a compound having a structure of Formula (I), (II), (III), (FV), (V), or (VI) with a candidate molecule under conditions in which the compound and the protein interact, and determining whether the amount of the compound that interacts with the protein is modulated relative to a control interaction between the compound and the protein without the candidate molecule, whereby a candidate molecule that modulates the amount of the compound interacting with the protein relative to the control interaction is identified as a candidate molecule that interacts with the protein.
• CK2 casein kinase 2
• a method for modulating the activity of a CK2 protein comprises contacting a system comprising the protein with a compound having a structure of Formula (I), (II), (III), (IV), (V), or (VI) in an amount effective for modulating the activity of the protein.
• F6 The method of any one of embodiments F1-F5, wherein the protein, the compound or the molecule is in association with a solid phase.
• a method for inhibiting cell proliferation comprises contacting cells with a compound having a structure of Formula (I), (H), (III), (IV), (V), or (VI) in an amount effective to inhibit proliferation of the cells.
• G2 The method of embodiment Gl, wherein the cells are in a cancer cell line.
• cancer cell line is a breast cancer, prostate cancer, pancreatic cancer, lung cancer, hemopoietic cancer, colorectal cancer, skin cancer, or ovary cancer cell line.
• G5 The method of any one of embodiments G1-G4, wherein contacting cells with a compound having a structure of Formula (I), (II), (III), (IV), (V), or (VI) induces cell apoptosis.
• a method for treating a condition related to aberrant cell proliferation comprises administering a compound having a structure of Formula (I), (II), (III), (IV), (V), or (VI) to a subject in need thereof in an amount effective to treat the cell proliferative condition.
• a method for identifying a candidate molecule that interacts with a protein kinase which comprises: contacting a composition containing a protein kinase and a compound having a structure of Formula (I), (II), (III), (IV), (V), or (VI) under conditions in which the compound and the protein interact with a candidate molecule, and determining whether the amount of the compound that interacts with the protein kinase is modulated relative to a control interaction between the compound and the protein kinase without the candidate molecule, whereby a candidate molecule that modulates the amount of the compound interacting with the protein kinase relative to the control interaction is identified as a candidate molecule that interacts with the protein kinase.
• J4 The method of embodiment Jl , J2, or J3, wherein the protein kinase is a human serine-threonine protein kinase selected from the group consisting of CK2, CK2 ⁇ 2, PIM 1 , PIM2, PIM3, CDKl/cyclinB, c-RAF, Mer, MELK, HIPK3, HIPK2 and ZIPK.
• the protein kinase is a human serine-threonine protein kinase selected from the group consisting of CK2, CK2 ⁇ 2, PIM 1 , PIM2, PIM3, CDKl/cyclinB, c-RAF, Mer, MELK, HIPK3, HIPK2 and ZIPK.
• J5. The method of embodiment J2, wherein the protein kinase is a serine-threonine protein kinase which contains one or more of the following amino acids at positions corresponding to those listed in human CK2: leucine at position 45, methionine at position 163 and isoleucine at position 174.
• J6. The method of embodiment J5, wherein the serine threonine protein kinase is selected from the group consisting of CK2, STKlO, HIPK2, HIPK3, DAPK3, DYK2 and PIM-I .
• J7 The method of embodiment J 1 , J2, or J3, wherein the protein kinase is a human tyrosine protein kinase selected from the group consisting of FLTl , FLT2, FLT3, FLT3 (D835Y), and FLT4.
• the protein kinase is a human tyrosine protein kinase selected from the group consisting of FLTl , FLT2, FLT3, FLT3 (D835Y), and FLT4.
• JlO The method of any one of embodiments J1-J9, wherein the protein kinase activity is the transfer of a gamma phosphate from adenosine triphosphate to a peptide or protein substrate.
• a method for modulating a protein kinase activity which comprises contacting a system comprising a protein kinase protein with a compound of Formula (I), (II), (III), (IV), (V), or (VI) in an amount effective for modulating the activity of the protein kinase.
• the protein kinase is a human serine- threonine protein kinase selected from the group consisting of CK2, CK2 ⁇ 2, PIMl, PIM2, PIM3, CDKl/cyclinB, c-RAF, Mer, MELK, HIPK3, HIPK2 and ZIPK.
• K5. The method of embodiment K2, wherein the protein kinase is a human tyrosine protein kinase selected from the group consisting of FLTl , FLT2, FLT3, FLT3 (D835Y), and FLT4.
• a method for treating pain or inflammation in a subject which comprises administering a compound of Formula (I), (II), (III), (IV), (V), or (VI) to a subject in need thereof in an amount effective to treat the pain or the inflammation.
• a method for inhibiting angiogenesis in a subject which comprises administering a compound of Formula (I), (II), (III), (IV), (V), or (VI) to a subject in need thereof in an amount effective to inhibit the angiogenesis.
• composition of matter comprising a protein kinase and a compound of Formula (I), (II), (UI), (IV) 1 (V), or (VI).
• composition of embodiment Nl wherein the protein kinase is a human serine-threonine kinase or a human tyrosine kinase.
• a compound of Formula (I), (II), (HI), (IV), (V), or (VI) which is a compound in one of the Tables provided herein or a pharmaceutically acceptable salt thereof.

Priority Applications (1)

Applications Claiming Priority (2)

Publications (2)

Family

ID=40523789

Family Applications (1)

Country Status (3)

Cited By (3)

Families Citing this family (6)

Citations (4)

Family Cites Families (5)

• 2008
  • 2008-08-29 CA CA2698463A patent/CA2698463A1/en not_active Abandoned
  • 2008-08-29 WO PCT/US2008/074820 patent/WO2009097014A2/en active Application Filing
  • 2008-08-29 US US12/201,892 patent/US7910600B2/en active Active

Patent Citations (4)

Non-Patent Citations (1)

Also Published As

Similar Documents

Legal Events