Therapeutic Antibodies – Mustapha Haddach, Joe A. TRAN, Fabrice Pierre, Collin F. REGAN, Nicholas Raffaele, Suchitra Ravula, David M. RYCKMAN, Senhwa Biosciences Inc
Abstract for “Pyrazolopyrimidines, and related heterocycles, as CK2 inhibitors.”
“The invention contains compounds that inhibit protein kinase CK2 (CK2 activ) activity, and compositions containing these compounds. These compositions and compounds are useful in treating proliferative conditions such as cancer and other kinase-associated disorders like inflammation, pain and certain immunological disorders.
Background for “Pyrazolopyrimidines, and related heterocycles, as CK2 inhibitors.”
“Protein kinase CK2 (formerly known as Casein kinase II) is referred to herein under the name?CK2? It is a highly conserved and ubiquitous protein serine/threonine-kinase kinase. Two catalytic (alpha/or alpha) subunits are typically required to contain the holoenzyme in tetrameric complexes. subunits, and two regulatory subunits (beta). CK2 participates in a variety of cellular functions, including maintaining cell viability. Normal cells have a tightly controlled level of CK2 and this has been long considered to play a key role in cell proliferation and growth. The PCT/US2007/077464 and PCT/US2008/074820 describe the use of CK2 inhibitors to treat certain types of cancer.
“CK2’s importance and prevalence, as well as its evolutionary analysis, suggests that it is an ancient enzyme. This may explain why its longevity has made it so important in so many biochemical processes. Infectious pathogens such as viruses and protozoa have used CK2 from hosts as an integral part their survival and life cycle biochemical system. These characteristics are also why inhibitors for CK2 may be helpful in a wide range of medical treatments, as we will discuss. Curr. Med. Chem. 2008, 15:1870-1886. Inhibitors of CK2 including the compounds discussed herein should be helpful in treating a wide range of diseases and disorders.”
Cancerous cells have an elevated level of CK2, and evidence has shown that CK2 can suppress apoptosis by protecting regulatory proteins against caspase-mediated destruction. CK2’s anti-apoptotic activity may be responsible for its ability to play a role in tumorigenesis and transformation. CK2 has been associated with multiple myeloma, chronic and acute myelogenous lymphoma, and lymphoma. Additionally, CK2 activity is increased in solid tumors of breast, colon, rectum, and breast. A small molecule that inhibits CK2 is believed to cause apoptosis in pancreatic cancer cells and hepatocellular carcinoma cell lines (HegG2, Hep3, HeLa) and CK2 inhibitors significantly increased the resistance of RMS (Rhabdomyosarcoma), tumors towards TRAIL-induced apoptosis. An inhibitor of CK2 in combination with TRAIL, or a ligand to the TRAIL receptor would be helpful in treating RMS, which is the most common soft-tissue sarcoma among children. Additionally, high CK2 levels are strongly associated with aggressiveness in neoplasias. Treatment with a CK2 inhibitor from the invention will reduce the tendency for benign lesions to become malignant or to metastasize.
“Unlike other kinases or signaling pathways where mutations can cause loss of regulatory control, CK2 activity levels increase more often than not due to activation level changes. Guerra and Issinger suggest that this could be due to regulation through aggregation. Activity levels are not well correlated with mRNA levels. Many cancers have shown excessive activity of CK2 including lung tumors and SCCHN tumors. Id.”
Increased CK2 activity was associated with higher malignancy in colorectal cancers. It has been shown that CK2 activity and expression can lead to increased nuclear levels of NFKappaB in breast carcinoma cells. Patients with AML or CML in blast crisis have CK2 activity that is significantly higher than in normal patients. This suggests that an inhibitor should be especially effective in these cases. Multiple myeloma cells survive on high levels of CK2, and inhibitors were cytotoxic to MM cell.
The literature clearly shows that inhibiting CK2 is associated with tumor cell growth. A CK2 inhibitor was shown to inhibit the growth of murine p190 cells. Its interaction with Bcr/Abl has been shown to play an important part in the proliferation of Bcr/Abl expressing cell lines. This suggests that inhibitors of CK2 might be useful in treating Bcr/Abl positive leukemias. It has been demonstrated that CK2 inhibitors can slow down the progression of skin papillomas and prostate cancer xenografts in mice and prolong the survival of transgenic mice that are prostate-promoters. Id.”
Recent research has highlighted the role of CK2 for non-cancer diseases. Guerra & Issinger. Curr. Med. Chem., 2008, 15:1870-1886. There is increasing evidence that CK2 plays a role in key diseases of the central nervous systems, such as Alzheimer’s and Parkinson’s. The progressive neurodegeneration associated with Alzheimer’s disease may be caused by selective CK2-mediated phosphorylation (or CK2) of tau proteins. Recent studies also suggest that CK2 may play a role in brain ischemia and memory impairment. This is likely due to CK2’s regulatory influence on the PI3K survival pathway.
It has been demonstrated that “CK2 is involved in modulating inflammatory disorders such as acute and chronic inflammatory pain, glomerulonephritis and autoimmune diseases like multiple sclerosis (MS), systemic lupus erythematosus and rheumatoid arthritis. It positively regulates function of the serotonin 5-HT3 channel, activates Heme oxygenase Type 2, and increases the activity of neuronal Nitric oxide synthase. When administered to spinal cord tissue before pain testing, a selective CK2 inhibitor was found to significantly reduce the pain response in mice. It phosphorylates synovial fluid IIA phospholipase B2 and modulates the secretion DEK, a nuclear DNA binding protein. This is a proinflammatory molecule that can be found in synovial liquid of juvenile arthritis patients. Inhibiting CK2 will likely stop the progression of inflammatory diseases such as those discussed here. The inhibitors described here have been proven to be effective in treating pain in animal models.
“Protein Kinase CK2 also plays a role in disorders affecting the vascular system such as hypoxia, laminar stress, and atherosclerosis. CK2 also plays a role in bone tissue and skeletal muscle disorders, including impaired insulin signaling, cardiomyocyte hypertrophy and bone tissue mineralization. One study found that inhibitors of CK2 could slow down angiogenesis induced in cultured cells by growth factor. In a retinopathy model, a CK2 antagonist combined with octreotide, a somatostatin analogue, reduced neovascular tufts. The CK2 inhibitors discussed herein could be used in conjunction with a somatostatin analogue to treat retinopathy.
“CK2 has been also shown to phosphorylate GSK and troponin; therefore, CK2 plays an important role in skeletal muscles and bone tissue physiology and can be linked to diseases affecting the muscle tissue.”
Evidence suggests that CK2 may also be involved in the life cycle regulation and development of protozoal parasites such as Theileria parva and Trypanosoma cruzi. Numerous studies have shown that CK2 plays an important role in the regulation of cellular motility and invasion of protozoan parasites. Infected hosts with Leishmania dovani, Herpetomonas muscarum muscarum and Plasmodium falciparum, activation or excess activity of CK2 have been demonstrated to occur. T. can be blocked by inhibition of CK2 cruzi.”
“CK2 also interacts with and/or phosphorylates viral proteins associated with human immune deficiency virus type 1(HIV-1), human papillomavirus, and herpes simplex virus. Human cytomegalovirus and hepatitis C viruses, Borna virus virus, adenovirus and coxsackieviruses, coronaviruses, influenza and varicella virus. CK2 activates HIV-1 reverse transcriptionase and proteases both in vitro as well as in vivo. It also promotes the pathogenicity for simian?human immunodeficiencyvirus (SHIV), a model of HIV. Inhibitors of CK2 can therefore reduce the pathogenic effects of a model HIV infection. CK2 also phosphorylates many proteins in herpes virus and other viruses. Some evidence suggests that viruses have adopted CK2 to phosphorylate their essential life-cycle proteins. The inhibition of CK2 should prevent the spread of viral infections. Viral infections rely on CK2 to maintain their lives.
“CK2 is a rare kinase inhibitor because of the variety of biological processes it affects. It is also different from other kinases in that it is constitutively active and can use ATP and GTP. It is found in many tumors and rapidly growing tissues and is elevated in them. CK2’s unique structural characteristics allow for the discovery of highly CK2-specific inhibitors. Many kinase inhibitors can affect multiple kinases which increases the chance of off-target effects and variability among individual subjects. CK2 is a highly attractive target for drug development. The invention provides highly effective inhibitors against CK2 that can be used to treat a wide range of diseases and disorders caused by excessive, aberrant, or undesired CK2 activity.
“Compounds from Formula I were found to be active both on CK2 and on one or more Pim protein. Compounds of Formula (II), and (II?) are now more active than previously thought. They are more active on CK2 and have less activity on Pim kinases, but they are also more active than compounds of Formula (II). It is thought that their physiological activity derives from their CK2 activity, although this theory is not binding.
“The invention provides novel formulae of Formula (II and (II). “The current invention provides novel compounds of Formula (II) and (II? These novel compounds, which are similar to Formula I, have a surprisingly higher activity on CK2 than the compounds of Formula I. They can also be used to treat conditions that are sensitive to CK2 inhibition, such as those discussed herein. Formula II compounds are useful for treating conditions that are mediated or associated with excess activity of CK2 and have a lower chance of off-target effects due to inhibition of other kinases.
“DISCLOSURE of the INVENTION”
“The present invention provides chemical compounds that have certain biological activities, including, but not limited to, inhibiting cell growth, inhibiting angiogenesis and modulating proteinkinase activities. These molecules are more selective for CK2 activity than other kinases and modulate protein kinase CK2(CK2) activity than compounds lacking the amine group in Formula (II?) or (II). These compounds have biological effects that include, but are not limited, inhibiting gamma phosphate transfers from ATP to a protein/peptide substrate, inhibiting growth, inducing cell apoptosis and inhibiting angiogenesis. In part, the present invention provides methods for creating novel chemical compounds and analogs thereof and methods for using these compounds. These compositions can be combined with other materials, such as therapeutic agents, and they are also provided.
“Compounds of the general formula (I) have been shown to inhibit Pim and CK2 (PCT/US2010/035657):”
“”
“The Formula I compounds inhibit Pim, CK2, and sometimes inhibit other kinases. It is advantageous to choose compounds that only inhibit one target enzyme or receptor for pharmaceutical use. This is because side effects from off-target biochemical effects could be unpredictable. The compounds of Formula (II and (II) are now known to be effective in inhibiting one primary target enzyme or receptor. These compounds, which are similar to Formula I, have high levels of CK2 activity and are often more potent than other compounds like Formula I. However, they are usually selective for CK2 over Pim kinases. Their selectivity for CK2 is higher than that of Formula I compounds. Compounds of Formula (II), or (II?) are therefore more useful than compounds of Formula I. Because they are selective for CK2 (or (II)), these compounds can be used to treat the conditions described in this article. They also inhibit fewer kinases so there is less chance of side effects.
“”
Formula (IIa or (IIa), are the most preferred compounds of Formula II. ):”
“”
“Particular embodiments include formula (II?Th) and (II?Th) thiophene-containing compound of the invention. ):”
“”
“The invention contains pharmaceutically acceptable salts compounds of Formula II, IIa and IIa?, IIa and IIa?. II-Th, II-Th, as well as II-Th. “, as well as neutral compounds.”
“The invention also includes pharmaceutical compositions that contain such compounds, as well as one or more pharmaceutically acceptable carriers/excipients. The invention also describes methods for using these compounds and compositions to treat specified conditions as further explained herein.”
“In addition, this invention provides intermediates for Formula (III), which can be used to prepare compounds described above. There are also methods of using these intermediaries to make compounds of Formula (II).
“”
“The method involves reacting a compound from Formula (III) and a hydantoin, or similar 5-membered heterocyclic compound from Formula (IV):
“”
“Typically, reaction conditions include a suitable solvent, a base and optionally a catalytic quantity of base. However, stoichiometric or higher amounts of base are possible. The bases that can deprotonate the compound of Formula IV to promote condensation with Formula (III), as well as secondary amines capable of reacting to aldehydes from Formula (III) to create an iminium species, are suitable bases. C1-C4 metal hydrides, C1-C4 alkoxides and tertiary aminos such as triethylamine and diisopropyl-ethylamine are all suitable bases. Piperidine, piperazine and N-methylpiperazine are also suitable secondary amine bases. Suitable solvents are polar aprotic solvents like NMP, DMF and DMSO as well as protic solvents like C1-C10 alcohols or diols such ethanol, propanol and isopropanol as well as ethylene glycol and propylene glycol and methoxyethanol. You can mix these solvents, or combine them with a less polar solvent to increase the solubility of the reactants. The ability to select suitable bases and solvents for these reactions is well within the reach of an average practitioner.
“In some formulations of Formula (III), the -L-W group represents a group from formula?S (O)1-2R, in which R is an alkyl or cycloalkyl and the product is a compound or (II?) The same -L?W group. These compounds can be used to prepare other compounds of formula (II or (II). Because the moiety in formula?S (O)1-2R can be easily displaced with nucleophiles like primary or secondary aminos, it is possible to introduce other -L?W groups. Another method of synthesizing compounds according to the invention is to react a compound from Formula (V),
“”
“”
“Also included herein are pharmaceutical compositions consisting of a compound from Formula I or II as defined herein, at least one pharmaceutically accepted carrier or excipient, and two or more pharmaceutically unacceptable carriers or excipients. These compositions can be used in treatment methods such as the ones described herein.
“The formulae I and II compounds described herein bind and inhibit certain kinase protein, which is thought to be their basis for pharmaceutical activity. The protein may be a CK2 if it is a CK2 CK2 protein. This could include a CK2 that contains the amino acid sequence SEQ ID NO: 1, 2, or 3, or a substantially similar variant thereof.
“(NP_001886;?casein?kinase?II?alpha?1?subunit?isoform?a?[Homo?sapiens])\nSEQ?ID?NO:?1\nmsgpvpsrar?vytdvnthrp?reywdyeshv?vewgnqddyq?lvrklgrgky?sevfeainit\nnnekvvvkil?kpvkkkkikr?eikilenlrg?gpniitladi?vkdpvsrtpa?lvfehvnntd\n121?fkqlyqtltd?ydirfymyei?lkaldychsm?gimhrdvkph?nvmidhehrk?lrlidwglae\n181?fyhpgqeynv?rvasryfkgp?ellvdyqmyd?ysldmwslgc?mlasmifrke?pffhghdnyd\n241?qlvriakvlg?tedlydyidk?ynieldprfn?dilgrhsrkr?werfvhsenq?hlvspealdf\n301?ldkllrydhq?srltareame?hpyfytvvkd?qarmgsssmp?ggstpvssan?mmsgissvpt\n361?psplgplags?pviaaanplg?mpvpaaagaq?q\n(NP_808227;?casein?kinase?II?alpha?1?subunit?isoform?a?[Homo?sapiens])\nSEQ?ID?NO:?2\nmsgpvpsrar?vytdvnthrp?reywdyeshv?vewgnqddyq?lvrklgrgky?sevfeainit\nnnekvvvkil?kpvkkkkikr?eikilenlrg?gpniitladi?vkdpvsrtpa?lvfehvnntd\n121?fkqlyqtltd?ydirfymyei?lkaldychsm?gimhrdvkph?nvmidhehrk?lrlidwglae\n181?fyhpgqeynv?rvasryfkgp?ellvdyqmyd?ysldmwslgc?mlasmifrke?pffhghdnyd\n241?qlvriakvlg?tedlydyidk?ynieldprfn?dilgrhsrkr?werfvhsenq?hlvspealdf\n301?ldkllrydhq?srltareame?hpyfytvvkd?qarmgsssmp?ggstpvssan?mmsgissvpt\n361?psplgplags?pviaaanplg?mpvpaaagaq?q\n(NP_808228;?casein?kinase?II?alpha?1?subunit?isoform?b? [Homo?sapiens])\nSEQ?ID?NO:?3\nmyeilkaldy?chsmgimhrd?vkphnvmidh?ehrklrlidw?glaefyhpgq?eynvrvasry\nfkgpellvdy?qmydysldmw?slgcmlasmi?frkepffhgh?dnydqlvria?kvlgtedlyd\n121?yidkynield?prfndilgrh?srkrwerfvh?senqhlvspe?aldfldkllr?ydhqsrltar\n181?eamehpyfyt?vvkdqarmgs?ssmpggstpv?ssanmmsgis?svptpsplgp?lagspviaaa\n241?nplgmpvpaa?agaqq”
These proteins are substitutially identical if they share at least 90% of their sequence homology, preferably at minimum 90% sequence identity. They also must have at least 50% of the in vitro kinase activity under standard assay conditions.
“The invention provides methods for modulating the activity of CK2 proteins, in vitro and ex vivo. One method is to contact the protein system with the compound described herein in a manner that modulates its activity. In some embodiments, the protein’s activity is reduced. Sometimes, the protein is a CK2 containing the amino acid sequence SEQID NO:1, SEQID NO:2 or 3 or a substantially identical variant thereof. In some embodiments, the CK2 is found in a cell or tissue. In other embodiments it can be present in a non-cell-free system.
“Provided are also methods to inhibit cell proliferation. These include contacting cells with the compound described in this document in an amount that inhibits proliferation. Sometimes, the cells are part of a cell line such as a cancer line. The cancer cell line may be a pancreatic, breast, or prostate cancer cell line in some embodiments. Sometimes the cells are found in tissue. At times they are in tumors. The method may also include inducing cell death. Sometimes cells are from someone with macular degeneration.
“Also, provided are methods to treat a condition related or aberrant cell proliferation. These include administering a compound herein to a subject who is in need of it in an amount that will effectively treat the cell proliferative disorder. The cell proliferative cancer may be a tumor-associated condition, such as a solid tumor or circulating tumor. Sometimes, the cancer is of the breast, prostate and pancreas. The cell proliferative state can be a non-tumorous cancer such as hematopoietic, including leukemias. In some embodiments, the cell proliferative condition may be macular degeneration.
“The invention also contains methods for treating cancer, an inflammatory disorder, or other disorders that are mediated through excessive activity of one or several of these kinases in a subject who is in need of such treatment. These include administering to a subject a therapeutically efficacious amount of a therapeutic drug useful for treating such disorder; administering to the patient a molecule herein, e.g., a molecule that inhibits CK2 in an effective amount to enhance the therapeutic agent’s. In some embodiments, the molecule which inhibits CK2 may be a compound of Formula I, Formula II, or Formula II. or (IIa or (IIa?) or (IIa) or (IIa? The desired effect of the therapeutic agents that inhibit CK2 in certain embodiments is an increase in apoptosis of at least one cell type. Certain embodiments require that the cell be a cancerous cell. The compound must be a compound of Formula (IIa or (IIa), which is a potent inhibitor (IC-50 lower than 100 nM for example) of CK2. The compound should have a IC-50 for Pim of less that 30 nM and be selective for CK2 over Pim kinases. In some embodiments, IC-50 to inhibit CK2 is lower than that for activity on Pim. Preferable embodiments have an IC-50 to CK2 that’s lower than its IC-50, which is for at least one Pim-1, Pim-2, and Pim-3, by around 100-fold or more.
“In some cases, the therapeutic agent is administered simultaneously with the molecule that inhibits CK2. Sometimes, the subject may use both the therapeutic agent and the molecule that inhibits CK2 simultaneously. In certain embodiments, the therapeutic agent and the molecule inhibiting CK2 may be combined to make one pharmaceutical composition. Other embodiments require that they are administered in separate compositions.
“Also included are compositions of matter consisting of a compound described herein as well as an isolated protein. Sometimes, the protein is a CK2 Protein, which may include a CK2 Protein that contains the amino acid sequence SEQID NO: 1, 2, or 3, or a substantially identical variant thereof. The protein may also be called a Pim protein in some instances. Some compositions include a compound described herein combined with a cell. A cell can be taken from a cell line such as a cancer-cell line. The cancer cell line in the latter embodiments may be a breast, prostate, pancreatic, lung, or skin cancer cell.
“These and other embodiments are described in this description.”
Summary for “Pyrazolopyrimidines, and related heterocycles, as CK2 inhibitors.”
“Protein kinase CK2 (formerly known as Casein kinase II) is referred to herein under the name?CK2? It is a highly conserved and ubiquitous protein serine/threonine-kinase kinase. Two catalytic (alpha/or alpha) subunits are typically required to contain the holoenzyme in tetrameric complexes. subunits, and two regulatory subunits (beta). CK2 participates in a variety of cellular functions, including maintaining cell viability. Normal cells have a tightly controlled level of CK2 and this has been long considered to play a key role in cell proliferation and growth. The PCT/US2007/077464 and PCT/US2008/074820 describe the use of CK2 inhibitors to treat certain types of cancer.
“CK2’s importance and prevalence, as well as its evolutionary analysis, suggests that it is an ancient enzyme. This may explain why its longevity has made it so important in so many biochemical processes. Infectious pathogens such as viruses and protozoa have used CK2 from hosts as an integral part their survival and life cycle biochemical system. These characteristics are also why inhibitors for CK2 may be helpful in a wide range of medical treatments, as we will discuss. Curr. Med. Chem. 2008, 15:1870-1886. Inhibitors of CK2 including the compounds discussed herein should be helpful in treating a wide range of diseases and disorders.”
Cancerous cells have an elevated level of CK2, and evidence has shown that CK2 can suppress apoptosis by protecting regulatory proteins against caspase-mediated destruction. CK2’s anti-apoptotic activity may be responsible for its ability to play a role in tumorigenesis and transformation. CK2 has been associated with multiple myeloma, chronic and acute myelogenous lymphoma, and lymphoma. Additionally, CK2 activity is increased in solid tumors of breast, colon, rectum, and breast. A small molecule that inhibits CK2 is believed to cause apoptosis in pancreatic cancer cells and hepatocellular carcinoma cell lines (HegG2, Hep3, HeLa) and CK2 inhibitors significantly increased the resistance of RMS (Rhabdomyosarcoma), tumors towards TRAIL-induced apoptosis. An inhibitor of CK2 in combination with TRAIL, or a ligand to the TRAIL receptor would be helpful in treating RMS, which is the most common soft-tissue sarcoma among children. Additionally, high CK2 levels are strongly associated with aggressiveness in neoplasias. Treatment with a CK2 inhibitor from the invention will reduce the tendency for benign lesions to become malignant or to metastasize.
“Unlike other kinases or signaling pathways where mutations can cause loss of regulatory control, CK2 activity levels increase more often than not due to activation level changes. Guerra and Issinger suggest that this could be due to regulation through aggregation. Activity levels are not well correlated with mRNA levels. Many cancers have shown excessive activity of CK2 including lung tumors and SCCHN tumors. Id.”
Increased CK2 activity was associated with higher malignancy in colorectal cancers. It has been shown that CK2 activity and expression can lead to increased nuclear levels of NFKappaB in breast carcinoma cells. Patients with AML or CML in blast crisis have CK2 activity that is significantly higher than in normal patients. This suggests that an inhibitor should be especially effective in these cases. Multiple myeloma cells survive on high levels of CK2, and inhibitors were cytotoxic to MM cell.
The literature clearly shows that inhibiting CK2 is associated with tumor cell growth. A CK2 inhibitor was shown to inhibit the growth of murine p190 cells. Its interaction with Bcr/Abl has been shown to play an important part in the proliferation of Bcr/Abl expressing cell lines. This suggests that inhibitors of CK2 might be useful in treating Bcr/Abl positive leukemias. It has been demonstrated that CK2 inhibitors can slow down the progression of skin papillomas and prostate cancer xenografts in mice and prolong the survival of transgenic mice that are prostate-promoters. Id.”
Recent research has highlighted the role of CK2 for non-cancer diseases. Guerra & Issinger. Curr. Med. Chem., 2008, 15:1870-1886. There is increasing evidence that CK2 plays a role in key diseases of the central nervous systems, such as Alzheimer’s and Parkinson’s. The progressive neurodegeneration associated with Alzheimer’s disease may be caused by selective CK2-mediated phosphorylation (or CK2) of tau proteins. Recent studies also suggest that CK2 may play a role in brain ischemia and memory impairment. This is likely due to CK2’s regulatory influence on the PI3K survival pathway.
It has been demonstrated that “CK2 is involved in modulating inflammatory disorders such as acute and chronic inflammatory pain, glomerulonephritis and autoimmune diseases like multiple sclerosis (MS), systemic lupus erythematosus and rheumatoid arthritis. It positively regulates function of the serotonin 5-HT3 channel, activates Heme oxygenase Type 2, and increases the activity of neuronal Nitric oxide synthase. When administered to spinal cord tissue before pain testing, a selective CK2 inhibitor was found to significantly reduce the pain response in mice. It phosphorylates synovial fluid IIA phospholipase B2 and modulates the secretion DEK, a nuclear DNA binding protein. This is a proinflammatory molecule that can be found in synovial liquid of juvenile arthritis patients. Inhibiting CK2 will likely stop the progression of inflammatory diseases such as those discussed here. The inhibitors described here have been proven to be effective in treating pain in animal models.
“Protein Kinase CK2 also plays a role in disorders affecting the vascular system such as hypoxia, laminar stress, and atherosclerosis. CK2 also plays a role in bone tissue and skeletal muscle disorders, including impaired insulin signaling, cardiomyocyte hypertrophy and bone tissue mineralization. One study found that inhibitors of CK2 could slow down angiogenesis induced in cultured cells by growth factor. In a retinopathy model, a CK2 antagonist combined with octreotide, a somatostatin analogue, reduced neovascular tufts. The CK2 inhibitors discussed herein could be used in conjunction with a somatostatin analogue to treat retinopathy.
“CK2 has been also shown to phosphorylate GSK and troponin; therefore, CK2 plays an important role in skeletal muscles and bone tissue physiology and can be linked to diseases affecting the muscle tissue.”
Evidence suggests that CK2 may also be involved in the life cycle regulation and development of protozoal parasites such as Theileria parva and Trypanosoma cruzi. Numerous studies have shown that CK2 plays an important role in the regulation of cellular motility and invasion of protozoan parasites. Infected hosts with Leishmania dovani, Herpetomonas muscarum muscarum and Plasmodium falciparum, activation or excess activity of CK2 have been demonstrated to occur. T. can be blocked by inhibition of CK2 cruzi.”
“CK2 also interacts with and/or phosphorylates viral proteins associated with human immune deficiency virus type 1(HIV-1), human papillomavirus, and herpes simplex virus. Human cytomegalovirus and hepatitis C viruses, Borna virus virus, adenovirus and coxsackieviruses, coronaviruses, influenza and varicella virus. CK2 activates HIV-1 reverse transcriptionase and proteases both in vitro as well as in vivo. It also promotes the pathogenicity for simian?human immunodeficiencyvirus (SHIV), a model of HIV. Inhibitors of CK2 can therefore reduce the pathogenic effects of a model HIV infection. CK2 also phosphorylates many proteins in herpes virus and other viruses. Some evidence suggests that viruses have adopted CK2 to phosphorylate their essential life-cycle proteins. The inhibition of CK2 should prevent the spread of viral infections. Viral infections rely on CK2 to maintain their lives.
“CK2 is a rare kinase inhibitor because of the variety of biological processes it affects. It is also different from other kinases in that it is constitutively active and can use ATP and GTP. It is found in many tumors and rapidly growing tissues and is elevated in them. CK2’s unique structural characteristics allow for the discovery of highly CK2-specific inhibitors. Many kinase inhibitors can affect multiple kinases which increases the chance of off-target effects and variability among individual subjects. CK2 is a highly attractive target for drug development. The invention provides highly effective inhibitors against CK2 that can be used to treat a wide range of diseases and disorders caused by excessive, aberrant, or undesired CK2 activity.
“Compounds from Formula I were found to be active both on CK2 and on one or more Pim protein. Compounds of Formula (II), and (II?) are now more active than previously thought. They are more active on CK2 and have less activity on Pim kinases, but they are also more active than compounds of Formula (II). It is thought that their physiological activity derives from their CK2 activity, although this theory is not binding.
“The invention provides novel formulae of Formula (II and (II). “The current invention provides novel compounds of Formula (II) and (II? These novel compounds, which are similar to Formula I, have a surprisingly higher activity on CK2 than the compounds of Formula I. They can also be used to treat conditions that are sensitive to CK2 inhibition, such as those discussed herein. Formula II compounds are useful for treating conditions that are mediated or associated with excess activity of CK2 and have a lower chance of off-target effects due to inhibition of other kinases.
“DISCLOSURE of the INVENTION”
“The present invention provides chemical compounds that have certain biological activities, including, but not limited to, inhibiting cell growth, inhibiting angiogenesis and modulating proteinkinase activities. These molecules are more selective for CK2 activity than other kinases and modulate protein kinase CK2(CK2) activity than compounds lacking the amine group in Formula (II?) or (II). These compounds have biological effects that include, but are not limited, inhibiting gamma phosphate transfers from ATP to a protein/peptide substrate, inhibiting growth, inducing cell apoptosis and inhibiting angiogenesis. In part, the present invention provides methods for creating novel chemical compounds and analogs thereof and methods for using these compounds. These compositions can be combined with other materials, such as therapeutic agents, and they are also provided.
“Compounds of the general formula (I) have been shown to inhibit Pim and CK2 (PCT/US2010/035657):”
“”
“The Formula I compounds inhibit Pim, CK2, and sometimes inhibit other kinases. It is advantageous to choose compounds that only inhibit one target enzyme or receptor for pharmaceutical use. This is because side effects from off-target biochemical effects could be unpredictable. The compounds of Formula (II and (II) are now known to be effective in inhibiting one primary target enzyme or receptor. These compounds, which are similar to Formula I, have high levels of CK2 activity and are often more potent than other compounds like Formula I. However, they are usually selective for CK2 over Pim kinases. Their selectivity for CK2 is higher than that of Formula I compounds. Compounds of Formula (II), or (II?) are therefore more useful than compounds of Formula I. Because they are selective for CK2 (or (II)), these compounds can be used to treat the conditions described in this article. They also inhibit fewer kinases so there is less chance of side effects.
“”
Formula (IIa or (IIa), are the most preferred compounds of Formula II. ):”
“”
“Particular embodiments include formula (II?Th) and (II?Th) thiophene-containing compound of the invention. ):”
“”
“The invention contains pharmaceutically acceptable salts compounds of Formula II, IIa and IIa?, IIa and IIa?. II-Th, II-Th, as well as II-Th. “, as well as neutral compounds.”
“The invention also includes pharmaceutical compositions that contain such compounds, as well as one or more pharmaceutically acceptable carriers/excipients. The invention also describes methods for using these compounds and compositions to treat specified conditions as further explained herein.”
“In addition, this invention provides intermediates for Formula (III), which can be used to prepare compounds described above. There are also methods of using these intermediaries to make compounds of Formula (II).
“”
“The method involves reacting a compound from Formula (III) and a hydantoin, or similar 5-membered heterocyclic compound from Formula (IV):
“”
“Typically, reaction conditions include a suitable solvent, a base and optionally a catalytic quantity of base. However, stoichiometric or higher amounts of base are possible. The bases that can deprotonate the compound of Formula IV to promote condensation with Formula (III), as well as secondary amines capable of reacting to aldehydes from Formula (III) to create an iminium species, are suitable bases. C1-C4 metal hydrides, C1-C4 alkoxides and tertiary aminos such as triethylamine and diisopropyl-ethylamine are all suitable bases. Piperidine, piperazine and N-methylpiperazine are also suitable secondary amine bases. Suitable solvents are polar aprotic solvents like NMP, DMF and DMSO as well as protic solvents like C1-C10 alcohols or diols such ethanol, propanol and isopropanol as well as ethylene glycol and propylene glycol and methoxyethanol. You can mix these solvents, or combine them with a less polar solvent to increase the solubility of the reactants. The ability to select suitable bases and solvents for these reactions is well within the reach of an average practitioner.
“In some formulations of Formula (III), the -L-W group represents a group from formula?S (O)1-2R, in which R is an alkyl or cycloalkyl and the product is a compound or (II?) The same -L?W group. These compounds can be used to prepare other compounds of formula (II or (II). Because the moiety in formula?S (O)1-2R can be easily displaced with nucleophiles like primary or secondary aminos, it is possible to introduce other -L?W groups. Another method of synthesizing compounds according to the invention is to react a compound from Formula (V),
“”
“”
“Also included herein are pharmaceutical compositions consisting of a compound from Formula I or II as defined herein, at least one pharmaceutically accepted carrier or excipient, and two or more pharmaceutically unacceptable carriers or excipients. These compositions can be used in treatment methods such as the ones described herein.
“The formulae I and II compounds described herein bind and inhibit certain kinase protein, which is thought to be their basis for pharmaceutical activity. The protein may be a CK2 if it is a CK2 CK2 protein. This could include a CK2 that contains the amino acid sequence SEQ ID NO: 1, 2, or 3, or a substantially similar variant thereof.
“(NP_001886;?casein?kinase?II?alpha?1?subunit?isoform?a?[Homo?sapiens])\nSEQ?ID?NO:?1\nmsgpvpsrar?vytdvnthrp?reywdyeshv?vewgnqddyq?lvrklgrgky?sevfeainit\nnnekvvvkil?kpvkkkkikr?eikilenlrg?gpniitladi?vkdpvsrtpa?lvfehvnntd\n121?fkqlyqtltd?ydirfymyei?lkaldychsm?gimhrdvkph?nvmidhehrk?lrlidwglae\n181?fyhpgqeynv?rvasryfkgp?ellvdyqmyd?ysldmwslgc?mlasmifrke?pffhghdnyd\n241?qlvriakvlg?tedlydyidk?ynieldprfn?dilgrhsrkr?werfvhsenq?hlvspealdf\n301?ldkllrydhq?srltareame?hpyfytvvkd?qarmgsssmp?ggstpvssan?mmsgissvpt\n361?psplgplags?pviaaanplg?mpvpaaagaq?q\n(NP_808227;?casein?kinase?II?alpha?1?subunit?isoform?a?[Homo?sapiens])\nSEQ?ID?NO:?2\nmsgpvpsrar?vytdvnthrp?reywdyeshv?vewgnqddyq?lvrklgrgky?sevfeainit\nnnekvvvkil?kpvkkkkikr?eikilenlrg?gpniitladi?vkdpvsrtpa?lvfehvnntd\n121?fkqlyqtltd?ydirfymyei?lkaldychsm?gimhrdvkph?nvmidhehrk?lrlidwglae\n181?fyhpgqeynv?rvasryfkgp?ellvdyqmyd?ysldmwslgc?mlasmifrke?pffhghdnyd\n241?qlvriakvlg?tedlydyidk?ynieldprfn?dilgrhsrkr?werfvhsenq?hlvspealdf\n301?ldkllrydhq?srltareame?hpyfytvvkd?qarmgsssmp?ggstpvssan?mmsgissvpt\n361?psplgplags?pviaaanplg?mpvpaaagaq?q\n(NP_808228;?casein?kinase?II?alpha?1?subunit?isoform?b? [Homo?sapiens])\nSEQ?ID?NO:?3\nmyeilkaldy?chsmgimhrd?vkphnvmidh?ehrklrlidw?glaefyhpgq?eynvrvasry\nfkgpellvdy?qmydysldmw?slgcmlasmi?frkepffhgh?dnydqlvria?kvlgtedlyd\n121?yidkynield?prfndilgrh?srkrwerfvh?senqhlvspe?aldfldkllr?ydhqsrltar\n181?eamehpyfyt?vvkdqarmgs?ssmpggstpv?ssanmmsgis?svptpsplgp?lagspviaaa\n241?nplgmpvpaa?agaqq”
These proteins are substitutially identical if they share at least 90% of their sequence homology, preferably at minimum 90% sequence identity. They also must have at least 50% of the in vitro kinase activity under standard assay conditions.
“The invention provides methods for modulating the activity of CK2 proteins, in vitro and ex vivo. One method is to contact the protein system with the compound described herein in a manner that modulates its activity. In some embodiments, the protein’s activity is reduced. Sometimes, the protein is a CK2 containing the amino acid sequence SEQID NO:1, SEQID NO:2 or 3 or a substantially identical variant thereof. In some embodiments, the CK2 is found in a cell or tissue. In other embodiments it can be present in a non-cell-free system.
“Provided are also methods to inhibit cell proliferation. These include contacting cells with the compound described in this document in an amount that inhibits proliferation. Sometimes, the cells are part of a cell line such as a cancer line. The cancer cell line may be a pancreatic, breast, or prostate cancer cell line in some embodiments. Sometimes the cells are found in tissue. At times they are in tumors. The method may also include inducing cell death. Sometimes cells are from someone with macular degeneration.
“Also, provided are methods to treat a condition related or aberrant cell proliferation. These include administering a compound herein to a subject who is in need of it in an amount that will effectively treat the cell proliferative disorder. The cell proliferative cancer may be a tumor-associated condition, such as a solid tumor or circulating tumor. Sometimes, the cancer is of the breast, prostate and pancreas. The cell proliferative state can be a non-tumorous cancer such as hematopoietic, including leukemias. In some embodiments, the cell proliferative condition may be macular degeneration.
“The invention also contains methods for treating cancer, an inflammatory disorder, or other disorders that are mediated through excessive activity of one or several of these kinases in a subject who is in need of such treatment. These include administering to a subject a therapeutically efficacious amount of a therapeutic drug useful for treating such disorder; administering to the patient a molecule herein, e.g., a molecule that inhibits CK2 in an effective amount to enhance the therapeutic agent’s. In some embodiments, the molecule which inhibits CK2 may be a compound of Formula I, Formula II, or Formula II. or (IIa or (IIa?) or (IIa) or (IIa? The desired effect of the therapeutic agents that inhibit CK2 in certain embodiments is an increase in apoptosis of at least one cell type. Certain embodiments require that the cell be a cancerous cell. The compound must be a compound of Formula (IIa or (IIa), which is a potent inhibitor (IC-50 lower than 100 nM for example) of CK2. The compound should have a IC-50 for Pim of less that 30 nM and be selective for CK2 over Pim kinases. In some embodiments, IC-50 to inhibit CK2 is lower than that for activity on Pim. Preferable embodiments have an IC-50 to CK2 that’s lower than its IC-50, which is for at least one Pim-1, Pim-2, and Pim-3, by around 100-fold or more.
“In some cases, the therapeutic agent is administered simultaneously with the molecule that inhibits CK2. Sometimes, the subject may use both the therapeutic agent and the molecule that inhibits CK2 simultaneously. In certain embodiments, the therapeutic agent and the molecule inhibiting CK2 may be combined to make one pharmaceutical composition. Other embodiments require that they are administered in separate compositions.
“Also included are compositions of matter consisting of a compound described herein as well as an isolated protein. Sometimes, the protein is a CK2 Protein, which may include a CK2 Protein that contains the amino acid sequence SEQID NO: 1, 2, or 3, or a substantially identical variant thereof. The protein may also be called a Pim protein in some instances. Some compositions include a compound described herein combined with a cell. A cell can be taken from a cell line such as a cancer-cell line. The cancer cell line in the latter embodiments may be a breast, prostate, pancreatic, lung, or skin cancer cell.
“These and other embodiments are described in this description.”
Click here to view the patent on Google Patents.