DIFFERENT ACTIVE SITES OF THE PROTEASOME AS DRUG TARGETS IN CANCER
Alexei Kisselev
Dartmouth College, Office Of Sponsored Projects, Hanover, Nh 03755
Grant 5R01CA124634-04 from National Cancer Institute
Abstract: Protein degradation by proteasomes plays an essential role in the proliferation of malignant cells. The proteasome inhibitor VELCADE (bortezomib, PS-341) is being used for the treatment of multiple myeloma and is in clinical trials for the treatment of other cancers. The proteasome has three types of active sites chymotrypsin-like (Beta 5), trypsin-like (Beta 2), and caspase-like (Beta 1). Our long-term goal is to understand the precise roles of these active sites as targets of anti-neoplastic agents and to use this information to design new, more potent, less toxic drugs of this class. Bortezomib was developed as an inhibitor of the Beta 5 site, which has long been considered the only suitable target for inhibition. However, bortezomib also inhibits the (Beta 1 site, and we have recently obtained data showing that inhibition of the Beta 1 site is needed to achieve maximal cytotoxicity. Based on these data and on critical review of the literature, we hypothesize that (a) all three active sites are drug molecular targets, (b) inhibition of at least two sites is required to achieve optimal cytotoxicity, (c) the therapeutic windows of proteasome inhibitors depend on which active sites they target, and (d) the exact pathways by which proteasome inhibitors induce apoptosis in malignant cells depend on which active sites they target. We will test this hypothesis in multiple myeloma and breast cancer cells using unique, cell-permeable, specific inhibitors of proteasome catalytic sites that we have developed and are developing. The specific aims of this proposal are as follows (1) To determine the growth inhibitory and cytotoxic effects of specific inhibitors of (Beta 5, Beta1, and Beta 2 catalytic sites, alone and in combination with each other, on cells derived from multiple myeloma and breast cancers, on immortalized and transformed human mammary epithelial cells, and on human peripheral blood lymphocytes; (2) To determine whether the mechanisms by which proteasome inhibitors kill multiple myeloma cells depend on the catalytic sites being targeted by these agents; The results of these studies will define what active-site pharmacological specificities proteasome inhibitors must have to achieve optimal anti-neoplastic activity and therapeutic windows. Taken together, these studies will provide a strong rationale for development of novel inhibitors with desired molecular pharmacological properties. Such compounds have potentially broad applicability for cancer therapy
Keywords: 20S Catalytic Proteasome; 20S Core Proteasome; 20S Proteasome; 20S Proteosome; APF-1; ATP-Dependent Proteolysis Factor 1; Active Sites; Address; Apoptosis; Apoptosis Pathway; Apoptotic; Bortezomib; Breast Cancer Cell; Breast Cancer Model; Cancer Treatment; Cancer cell line; Cancer of Breast; Cancers; Catalytic Core; Catalytic Domain; Catalytic Region; Catalytic Site; Catalytic Subunit; Cell Death, Programmed; Cell Growth in Number; Cell Line; Cell Lines, Strains; Cell Multiplication; Cell Proliferation; Cell-Death Protease; CellLine; Cells; Cellular Proliferation; Clinical Treatment; Clinical Trials; Clinical Trials, Phase I; Clinical Trials, Unspecified; Data; Development; Disease; Disorder; Drug Delivery; Drug Delivery Systems; Drug Targeting; Drug Targetings; Drugs; Early-Stage Clinical Trials; Epithelial Cells; Estrogen receptor negative; Figs; Figs - dietary; Generalized Growth; Goals; Growth; HMG-20; High Mobility Protein 20; Human; Human Breast Cancer Cell; Human, General; ICE-like protease; Ich-1 protein; Investigators; Killings; Lytotoxicity; Macropain; Macroxyproteinase; Malignant Cell; Malignant Neoplasm Therapy; Malignant Neoplasm Treatment; Malignant Neoplasms; Malignant Tumor; Malignant Tumor of the Breast; Malignant neoplasm of breast; Mammary Glands, Human; Mammary gland; Man (Taxonomy); Man, Modern; Medication; Molecular; Molecular Target; Multicatalytic Proteinase; Multiple Myeloma; Myeloma, Plasma-Cell; Nedd-2 protein; Non-Malignant; Oncogenesis; PBL; PBMC; Pathway interactions; Peripheral Blood Lymphocyte; Peripheral Blood Mononuclear Cell; Pharmaceutic Preparations; Pharmaceutical Preparations; Phase 1 Clinical Trials; Phase I Clinical Trials; Phase I Study; Play; Programs (PT); Programs [Publication Type]; Property; Property, LOINC Axis 2; Prosome; Proteasome; Proteasome Endopeptidase Complex; Proteasome Inhibitor; Protein Degradation, Metabolic; Protein Degradation, Regulatory; Protein Turnover; Proteins; Proteosome; Public Health; Quality Control; Research Personnel; Researchers; Resistance; Review Literature; Role; Site; Specificity; Staging; Structure; Testing; Therapeutic; Tissue Growth; Toxic effect; Toxicities; Tripcellim; Trypsin; Ubiquitin; Velcade; Work; anticancer therapy; base; cancer cell; cancer therapy; cancer type; caspase; caspase-2; chymotrypsin; clinical investigation; cultured cell line; cystein protease; cystein proteinase; cysteine endopeptidase; cytotoxic; cytotoxicity; design; designing; disease/disorder; drug/agent; gene product; inhibitor; inhibitor/antagonist; malignancy; malignant breast neoplasm; mammary; mammary cancer model; mammary tumor model; multicatalytic endopeptidase complex; myeloma; myelomatosis; neoplasm/cancer; neoplastic; nonmalignant; novel; ontogeny; pathway; phase 1 study; phase 1 trial; phase I trial; pre-clinical; preclinical; programs; protein degradation; protocol, phase I; public health medicine (field); resistant; salinosporamide A; social role; trial regimen; trial treatment; tumorigenesis
Project start date: 2007-07-20
Project end date: 2012-05-31
Budget start date: 1-JUN-2010
Budget end date: 31-MAY-2011
PFA/PA: PA-06-167
5R01CA124634-04 (2010): $273429
Sponsored Links Excellgen http://Excellgen.com
DIFFERENT ACTIVE SITES OF THE PROTEASOME AS DRUG TARGETS IN CANCER
Alexei Kisselev
Dartmouth College, Office Of Sponsored Projects, Hanover, Nh 03755
Grant 5R01CA124634-03 from National Cancer Institute
Abstract: Protein degradation by proteasomes plays an essential role in the proliferation of malignant cells. The proteasome inhibitor VELCADE (bortezomib, PS-341) is being used for the treatment of multiple myeloma and is in clinical trials for the treatment of other cancers. The proteasome has three types of active sites chymotrypsin-like (Beta 5), trypsin-like (Beta 2), and caspase-like (Beta 1). Our long-term goal is to understand the precise roles of these active sites as targets of anti-neoplastic agents and to use this information to design new, more potent, less toxic drugs of this class. Bortezomib was developed as an inhibitor of the Beta 5 site, which has long been considered the only suitable target for inhibition. However, bortezomib also inhibits the (Beta 1 site, and we have recently obtained data showing that inhibition of the Beta 1 site is needed to achieve maximal cytotoxicity. Based on these data and on critical review of the literature, we hypothesize that (a) all three active sites are drug molecular targets, (b) inhibition of at least two sites is required to achieve optimal cytotoxicity, (c) the therapeutic windows of proteasome inhibitors depend on which active sites they target, and (d) the exact pathways by which proteasome inhibitors induce apoptosis in malignant cells depend on which active sites they target. We will test this hypothesis in multiple myeloma and breast cancer cells using unique, cell-permeable, specific inhibitors of proteasome catalytic sites that we have developed and are developing. The specific aims of this proposal are as follows (1) To determine the growth inhibitory and cytotoxic effects of specific inhibitors of (Beta 5, Beta1, and Beta 2 catalytic sites, alone and in combination with each other, on cells derived from multiple myeloma and breast cancers, on immortalized and transformed human mammary epithelial cells, and on human peripheral blood lymphocytes; (2) To determine whether the mechanisms by which proteasome inhibitors kill multiple myeloma cells depend on the catalytic sites being targeted by these agents; The results of these studies will define what active-site pharmacological specificities proteasome inhibitors must have to achieve optimal anti-neoplastic activity and therapeutic windows. Taken together, these studies will provide a strong rationale for development of novel inhibitors with desired molecular pharmacological properties. Such compounds have potentially broad applicability for cancer therapy
Keywords: 20S Catalytic Proteasome; 20S Core Proteasome; 20S Proteasome; 20S Proteosome; APF-1; ATP-Dependent Proteolysis Factor 1; Active Sites; Address; Apoptosis; Apoptosis Pathway; Apoptotic; Bortezomib; Breast Cancer Cell; Breast Cancer Model; Cancer Treatment; Cancer cell line; Cancer of Breast; Cancers; Catalytic Core; Catalytic Domain; Catalytic Region; Catalytic Site; Catalytic Subunit; Cell Death, Programmed; Cell Growth in Number; Cell Line; Cell Lines, Strains; Cell Multiplication; Cell Proliferation; Cell-Death Protease; CellLine; Cells; Cellular Proliferation; Clinical Treatment; Clinical Trials; Clinical Trials, Phase I; Clinical Trials, Unspecified; Data; Development; Disease; Disorder; Drug Delivery; Drug Delivery Systems; Drug Targeting; Drug Targetings; Drugs; Early-Stage Clinical Trials; Epithelial Cells; Estrogen receptor negative; Figs; Figs - dietary; Generalized Growth; Goals; Growth; HMG-20; High Mobility Protein 20; Human; Human Breast Cancer Cell; Human, General; ICE-like protease; Ich-1 protein; Investigators; Killings; Lytotoxicity; Macropain; Macroxyproteinase; Malignant Cell; Malignant Neoplasm Therapy; Malignant Neoplasm Treatment; Malignant Neoplasms; Malignant Tumor; Malignant Tumor of the Breast; Malignant neoplasm of breast; Mammary Glands, Human; Mammary gland; Man (Taxonomy); Man, Modern; Medication; Molecular; Molecular Target; Multicatalytic Proteinase; Multiple Myeloma; Myeloma, Plasma-Cell; Nedd-2 protein; Non-Malignant; Oncogenesis; PBL; PBMC; Pathway interactions; Peripheral Blood Lymphocyte; Peripheral Blood Mononuclear Cell; Pharmaceutic Preparations; Pharmaceutical Preparations; Phase 1 Clinical Trials; Phase I Clinical Trials; Phase I Study; Play; Programs (PT); Programs [Publication Type]; Property; Property, LOINC Axis 2; Prosome; Proteasome; Proteasome Endopeptidase Complex; Proteasome Inhibitor; Protein Degradation, Metabolic; Protein Degradation, Regulatory; Protein Turnover; Proteins; Proteosome; Public Health; Quality Control; Research Personnel; Researchers; Resistance; Review Literature; Role; Site; Sites, Active; Specificity; Staging; Structure; Testing; Therapeutic; Tissue Growth; Toxic effect; Toxicities; Tripcellim; Trypsin; Ubiquitin; Velcade; Work; anticancer therapy; base; cancer cell; cancer therapy; cancer type; caspase; caspase-2; chymotrypsin; clinical investigation; cultured cell line; cystein protease; cystein proteinase; cysteine endopeptidase; cytotoxic; cytotoxicity; design; designing; disease/disorder; drug/agent; gene product; inhibitor; inhibitor/antagonist; malignancy; malignant breast neoplasm; mammary; mammary cancer model; mammary tumor model; multicatalytic endopeptidase complex; myeloma; myelomatosis; neoplasm/cancer; neoplastic; nonmalignant; novel; ontogeny; pathway; phase 1 study; phase 1 trial; phase I trial; pre-clinical; preclinical; programs; protein degradation; protocol, phase I; public health medicine (field); resistant; salinosporamide A; social role; trial regimen; trial treatment; tumorigenesis
Project start date: 2007-07-20
Project end date: 2012-05-31
Budget start date: 1-JUN-2009
Budget end date: 31-MAY-2010
PFA/PA: PA-06-167
5R01CA124634-03 (2009): $273429
Grants awarded to Alexei Kisselev
DIFFERENT ACTIVE SITES OF THE PROTEASOME AS DRUG TARGETS IN CANCER
Alexei Kisselev
Dartmouth College, Office Of Sponsored Projects, Hanover, Nh 03755
Grant 3R01CA124634-03S1 from National Cancer Institute
Abstract: Protein degradation by proteasomes plays an essential role in the proliferation of malignant cells. The proteasome inhibitor VELCADE (bortezomib, PS-341) is being used for the treatment of multiple myeloma and is in clinical trials for the treatment of other cancers. The proteasome has three types of active sites chymotrypsin-like (Beta 5), trypsin-like (Beta 2), and caspase-like (Beta 1). Our long-term goal is to understand the precise roles of these active sites as targets of anti-neoplastic agents and to use this information to design new, more potent, less toxic drugs of this class. Bortezomib was developed as an inhibitor of the Beta 5 site, which has long been considered the only suitable target for inhibition. However, bortezomib also inhibits the (Beta 1 site, and we have recently obtained data showing that inhibition of the Beta 1 site is needed to achieve maximal cytotoxicity. Based on these data and on critical review of the literature, we hypothesize that (a) all three active sites are drug molecular targets, (b) inhibition of at least two sites is required to achieve optimal cytotoxicity, (c) the therapeutic windows of proteasome inhibitors depend on which active sites they target, and (d) the exact pathways by which proteasome inhibitors induce apoptosis in malignant cells depend on which active sites they target. We will test this hypothesis in multiple myeloma and breast cancer cells using unique, cell-permeable, specific inhibitors of proteasome catalytic sites that we have developed and are developing. The specific aims of this proposal are as follows (1) To determine the growth inhibitory and cytotoxic effects of specific inhibitors of (Beta 5, Beta1, and Beta 2 catalytic sites, alone and in combination with each other, on cells derived from multiple myeloma and breast cancers, on immortalized and transformed human mammary epithelial cells, and on human peripheral blood lymphocytes; (2) To determine whether the mechanisms by which proteasome inhibitors kill multiple myeloma cells depend on the catalytic sites being targeted by these agents; The results of these studies will define what active-site pharmacological specificities proteasome inhibitors must have to achieve optimal anti-neoplastic activity and therapeutic windows. Taken together, these studies will provide a strong rationale for development of novel inhibitors with desired molecular pharmacological properties. Such compounds have potentially broad applicability for cancer therapy
Keywords: 20S Catalytic Proteasome; 20S Core Proteasome; 20S Proteasome; 20S Proteosome; APF-1; ATP-Dependent Proteolysis Factor 1; Active Sites; Address; Apoptosis; Apoptosis Pathway; Apoptotic; Bortezomib; Breast Cancer Cell; Breast Cancer Model; Cancer Treatment; Cancer cell line; Cancer of Breast; Cancers; Catalytic Core; Catalytic Domain; Catalytic Region; Catalytic Site; Catalytic Subunit; Cell Death, Programmed; Cell Growth in Number; Cell Line; Cell Lines, Strains; Cell Multiplication; Cell Proliferation; Cell-Death Protease; CellLine; Cells; Cellular Proliferation; Clinical Treatment; Clinical Trials; Clinical Trials, Phase I; Clinical Trials, Unspecified; Data; Development; Disease; Disorder; Drug Delivery; Drug Delivery Systems; Drug Targeting; Drug Targetings; Drugs; Early-Stage Clinical Trials; Epithelial Cells; Estrogen receptor negative; Figs; Figs - dietary; Generalized Growth; Goals; Growth; HMG-20; High Mobility Protein 20; Human; Human Breast Cancer Cell; Human, General; ICE-like protease; Ich-1 protein; Investigators; Killings; Lytotoxicity; Macropain; Macroxyproteinase; Malignant Cell; Malignant Neoplasm Therapy; Malignant Neoplasm Treatment; Malignant Neoplasms; Malignant Tumor; Malignant Tumor of the Breast; Malignant neoplasm of breast; Mammary Glands, Human; Mammary gland; Man (Taxonomy); Man, Modern; Medication; Molecular; Molecular Target; Multicatalytic Proteinase; Multiple Myeloma; Myeloma, Plasma-Cell; Nedd-2 protein; Non-Malignant; Oncogenesis; PBL; PBMC; Pathway interactions; Peripheral Blood Lymphocyte; Peripheral Blood Mononuclear Cell; Pharmaceutic Preparations; Pharmaceutical Preparations; Phase 1 Clinical Trials; Phase I Clinical Trials; Phase I Study; Play; Programs (PT); Programs [Publication Type]; Property; Property, LOINC Axis 2; Prosome; Proteasome; Proteasome Endopeptidase Complex; Proteasome Inhibitor; Protein Degradation, Metabolic; Protein Degradation, Regulatory; Protein Turnover; Proteins; Proteosome; Public Health; Quality Control; Research Personnel; Researchers; Resistance; Review Literature; Role; Site; Specificity; Staging; Structure; Testing; Therapeutic; Tissue Growth; Toxic effect; Toxicities; Tripcellim; Trypsin; Ubiquitin; Velcade; Work; anticancer therapy; base; cancer cell; cancer therapy; cancer type; caspase; caspase-2; chymotrypsin; clinical investigation; cultured cell line; cystein protease; cystein proteinase; cysteine endopeptidase; cytotoxic; cytotoxicity; design; designing; disease/disorder; drug/agent; gene product; inhibitor; inhibitor/antagonist; malignancy; malignant breast neoplasm; mammary; mammary cancer model; mammary tumor model; multicatalytic endopeptidase complex; myeloma; myelomatosis; neoplasm/cancer; neoplastic; nonmalignant; novel; ontogeny; pathway; phase 1 study; phase 1 trial; phase I trial; pre-clinical; preclinical; programs; protein degradation; protocol, phase I; public health medicine (field); resistant; salinosporamide A; social role; trial regimen; trial treatment; tumorigenesis
Project start date: 2009-08-01
Project end date: 2011-07-31
Budget start date: 1-AUG-2009
Budget end date: 31-JUL-2011
PFA/PA: PA-06-167
3R01CA124634-03S1 (2009): $270180