Clinical activity of carfilzomib correlates with inhibition of multiple proteasome subunits: application of a novel pharmacodynamic assay
Susan J Lee, Konstantin Levitsky, Francesco Parlati, Mark K Bennett, Shirin Arastu-Kapur, Lois Kellerman, Tina F Woo, Alvin F Wong, Kyriakos P Papadopoulos, Ruben Niesvizky, Ashraf Z Badros, Ravi Vij, Sundar Jagannath, David Siegel, Michael Wang, Gregory J Ahmann, Christopher J Kirk, Susan J Lee, Konstantin Levitsky, Francesco Parlati, Mark K Bennett, Shirin Arastu-Kapur, Lois Kellerman, Tina F Woo, Alvin F Wong, Kyriakos P Papadopoulos, Ruben Niesvizky, Ashraf Z Badros, Ravi Vij, Sundar Jagannath, David Siegel, Michael Wang, Gregory J Ahmann, Christopher J Kirk
Abstract
While proteasome inhibition is a validated therapeutic approach for multiple myeloma (MM), inhibition of individual constitutive proteasome (c20S) and immunoproteasome (i20S) subunits has not been fully explored owing to a lack of effective tools. We utilized the novel proteasome constitutive/immunoproteasome subunit enzyme-linked immunosorbent (ProCISE) assay to quantify proteasome subunit occupancy in samples from five phase I/II and II trials before and after treatment with the proteasome inhibitor carfilzomib. Following the first carfilzomib dose (15-56 mg/m(2) ), dose-dependent inhibition of c20S and i20S chymotrypsin-like active sites was observed [whole blood: ≥67%; peripheral blood mononuclear cells (PBMCs): ≥75%]. A similar inhibition profile was observed in bone marrow-derived CD138(+) tumour cells. Carfilzomib-induced proteasome inhibition was durable, with minimal recovery in PBMCs after 24 h but near-complete recovery between cycles. Importantly, the ProCISE assay can be used to quantify occupancy of individual c20S and i20S subunits. We observed a relationship between MM patient response (n = 29), carfilzomib dose and occupancy of multiple i20S subunits, where greater occupancy was associated with an increased likelihood of achieving a clinical response at higher doses. ProCISE represents a new tool for measuring proteasome inhibitor activity in clinical trials and relating drug action to patient outcomes.
Keywords: molecular analysis; multiple myeloma; myeloma therapy; pharmacology; trials.
© 2016 The Authors. British Journal of Haematology published by John Wiley & Sons Ltd.
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References
- Arastu‐Kapur, S. , Anderl, J.L. , Kraus, M. , Parlati, F. , Shenk, K.D. , Lee, S.J. , Muchamuel, T. , Bennett, M.K. , Driessen, C. , Ball, A.J. & Kirk, C.J. (2011) Nonproteasomal targets of the proteasome inhibitors bortezomib and carfilzomib: a link to clinical adverse events. Clinical Cancer Research, 17, 2734–2743.
- Badros, A.Z. , Vij, R. , Martin, T. , Zonder, J.A. , Kunkel, L. , Wang, Z. , Lee, S. , Wong, A.F. & Niesvizky, R. (2013) Carfilzomib in multiple myeloma patients with renal impairment: pharmacokinetics and safety. Leukemia, 27, 1707–1714.
- Bennett, M.K. & Kirk, C.J. (2008) Development of proteasome inhibitors in oncology and autoimmune diseases. Current Opinion in Drug Discovery and Development, 11, 616–625.
- Bennett, M.K. , Buchholz, T.J. , Demo, S.D. , Laidig, G.J. , Lewis, E.R. & Smyth, M.S. (2006) Compound for enzyme inhibition (US Patent 2006/0088471 A1), April 27, 2006. . Accessed January 16, 2015
- Britton, M. , Lucas, M.M. , Downey, S.L. , Screen, M. , Pletnev, A.A. , Verdoes, M. , Tokhunts, R.A. , Amir, O. , Goddard, A.L. , Pelphrey, P.M. , Wright, D.L. , Overkleeft, H.S. & Kisselev, A.F. (2009) Selective inhibitor of proteasome's caspase‐like sites sensitizes cells to specific inhibition of chymotrypsin‐like sites. Chemistry and Biology, 16, 1278–1289.
- Chauhan, D. , Catley, L. , Li, G. , Podar, K. , Hideshima, T. , Velankar, M. , Mitsiades, C. , Mitsiades, N. , Yasui, H. , Letai, A. , Ovaa, H. , Berkers, C. , Nicholson, B.l , Chao, T.‐H. , Neuteboom, S.T.C. , Richardson, P. , Palladino, M.A. & Anderson, K.C. (2005) A novel orally active proteasome inhibitor induces apoptosis in multiple myeloma cells with mechanisms distinct from bortezomib. Cancer Cell, 8, 407–419.
- Ciechanover, A. (2012) Intracellular protein degradation: from a vague idea thru the lysosome and the ubiquitin‐proteasome system and onto human diseases and drug targeting. Biochimica et Biophysica Acta, 1824, 3–13.
- Coux, O. , Tanaka, K. & Goldberg, A.L. (1996) Structure and functions of the 20S and 26S proteasomes. Annual Review of Biochemistry, 65, 801–847.
- Demo, S.D. , Kirk, C.J. , Aujay, M.A. , Buchholz, T.J. , Dajee, M. , Ho, M.N. , Jiang, J. , Laidig, G.J. , Lewis, E.R. , Parlati, F. , Shenk, K.D. , Smyth, M.S. , Sun, C.M. , Vallone, M.K. , Woo, T.M. , Molineaux, C.J. & Bennett, M.K. (2007) Antitumor activity of PR‐171, a novel irreversible inhibitor of the proteasome. Cancer Research, 67, 6383–6391.
- Dick, L.R. & Fleming, P.E. (2010) Building on bortezomib: second‐generation proteasome inhibitors as anti‐cancer therapy. Drug Discovery Today, 15, 243–249.
- Fisher, R.I. , Bernstein, S.H. , Kahl, B.S. , Djulbegovic, B. , Robertson, M.J. , de Vos, S. , Epner, E. , Krishnan, A. , Leonard, J.P. , Lonial, S. , Stadtmauer, E.A. , O'Connor, O.A. , Shi, H. , Boral, A.L. & Goy, A. (2006) Multicenter phase II study of bortezomib in patients with relapsed or refractory mantle cell lymphoma. Journal of Clinical Oncology, 24, 4867–4874.
- Glynne, R. , Powis, S.H. , Beck, S. , Kelly, A. , Kerr, L.A. & Trowsdale, J. (1991) A proteasome‐related gene between the two ABC transporter loci in the class II region of the human MHC. Nature, 353, 357–360.
- Groettrup, M. , Kirk, C.J. & Basler, M. (2010) Proteasomes in immune cells: more than peptide producers? Nature Reviews Immunology, 10, 73–78.
- Hellmann, A. , Rule, S. , Walewski, J. , Shpilberg, O. , Feng, H. , van de Velde, H. , Patel, H. , Skee, D.M. , Girgis, S. & Louw, V.J. (2011) Effect of cytochrome P450 3A4 inducers on the pharmacokinetic, pharmacodynamic and safety profiles of bortezomib in patients with multiple myeloma or non‐Hodgkin's lymphoma. Clinical Pharmacokinetics, 50, 781–791.
- Jagannath, S. , Barlogie, B. , Berenson, J.R. , Singhal, S. , Alexanian, R. , Srkalovic, G. , Orlowski, R.Z. , Richardson, P.G. , Anderson, J.A. , Nix, D. , Esseltine, D.L. & Anderson, K.C. (2005) Bortezomib in recurrent and/or refractory multiple myeloma. Initial clinical experience in patients with impaired renal function. Cancer, 103, 1195–1200.
- Jain, S. , Diefenbach, C. , Zain, J. & O'Connor, O.A. (2011) Emerging role of carfilzomib in treatment of relapsed and refractory lymphoid neoplasms and multiple myeloma. Core Evidence, 6, 43–57.
- Kraus, M. , Rückrich, T. , Reich, M. , Gogel, J. , Beck, A. , Kammer, W. , Berkers, C.R. , Burg, D. , Overkleeft, H. , Ovaa, H. & Driessen, C. (2007) Activity patterns of proteasome subunits reflect bortezomib sensitivity of hematologic malignancies and are variable in primary human leukemia cells. Leukemia, 21, 84–92.
- Kuhn, D.J. , Chen, Q. , Voorhees, P.M. , Strader, J.S. , Shenk, K.D. , Sun, C.M. , Demo, S.D. , Bennett, M.K. , van Leeuwen, F.W.B. , Chanan‐Khan, A.A. & Orlowski, R.Z. (2007) Potent activity of carfilzomib, a novel, irreversible inhibitor of the ubiquitin‐proteasome pathway, against preclinical models of multiple myeloma. Blood, 110, 3281–3290.
- Kuhn, D.J. , Hunsucker, S.A. , Chen, Q. , Voorhees, P.M. , Orlowski, M. & Orlowski, R.Z. (2009) Targeted inhibition of the immunoproteasome is a potent strategy against models of multiple myeloma that overcomes resistance to conventional drugs and nonspecific proteasome inhibitors. Blood, 113, 4667–4676.
- Kuhn, D.J. , Orlowski, R.Z. & Bjorklund, C.C. (2011) Second generation proteasome inhibitors: carfilzomib and immunoproteasome‐specific inhibitors (IPSIs). Current Cancer Drug Targets, 11, 285–295.
- Kummar, S. , Chen, H.X. , Wright, J. , Holbeck, S. , Millin, M.D. , Tomaszewski, J. , Zweibel, J. , Collins, J. & Doroshow, J.H. (2010) Utilizing targeted cancer therapeutic agents in combination: novel approaches and urgent requirements. Nature Reviews Drug Discovery, 9, 843–856.
- Lightcap, E.S. , McCormack, T.A. , Pien, C.S. , Chau, V. , Adams, J. & Elliott, P.J. (2000) Proteasome inhibition measurements: clinical application. Clinical Chemistry, 46, 673–683.
- Martinez, C.K. & Monaco, J.J. (1991) Homology of proteasome subunits to a major histocompatibility complex‐linked LMP gene. Nature 353, 664–667.
- Mirabella, A.C. , Pletnev, A.A. , Downey, S.L. , Florea, B.I. , Shabaneh, T.B. , Britton, M. , Verdoes, M. , Filippov, D.V. , Overkleeft, H.S. & Kisselev, A.F. (2011) Specific cell‐permeable inhibitor of proteasome trypsin‐like sites selectively sensitizes myeloma cells to bortezomib and carfilzomib. Chemistry and Biology, 18, 608–618.
- Moreau, P. , Coiteux, V. , Hulin, C. , Leleu, X. , van de Velde, H. , Acharya, M. & Harousseau, J.‐L. (2008) Prospective comparison of subcutaneous versus intravenous administration of bortezomib in patients with multiple myeloma. Haematologica, 93, 1908–1911.
- Muchamuel, T. , Basler, M. , Aujay, M.A. , Suzuki, E. , Kalim, K.W. , Lauer, C. , Sylvain, C. , Ring, E.R. , Shields, J. , Jiang, J. , Shwonek, Pl , Parlati, F. , Demo, S.D. , Bennett, M.K. , Kirk, C.J. & Groettrup, M. (2009) A selective inhibitor of the immunoproteasome subunit LMP7 blocks cytokine production and attenuates progression of experimental arthritis. Nature Medicine, 15, 781–787.
- Nandi, D. , Jiang, H. & Monaco, J.J. (1996) Identification of MECL‐1 (LMP‐10) as the third IFN‐gamma‐inducible proteasome subunit. Journal of Immunology, 156, 2361–2364.
- Niesvizky, R. , Martin, T.G. III , Bensinger, W.I. , Alsina, M. , Siegel, D.S. , Kunkel, L.A. , Wong, A.F. , Lee, S. , Orlowski, R.Z. & Wang, M. (2013) Phase Ib dose‐escalation study (PX‐171‐006) of carfilzomib, lenalidomide, and low‐dose dexamethasone in relapsed or progressive multiple myeloma. Clinical Cancer Research, 19, 2248–2256.
- O'Connor, O.A. , Stewart, A.K. , Vallone, M. , Molineaux, C.J. , Kunkel, L.A. , Gerecitano, J.F. & Orlowski, R.Z. (2009) A phase 1 dose escalation study of the safety and pharmacokinetics of the novel proteasome inhibitor carfilzomib (PR‐171) in patients with hematologic malignancies. Clinical Cancer Research, 15, 7085–7091.
- Orlowski, R.Z. , Stinchcombe, T.E. , Mitchell, B.S. , Shea, T.C. , Baldwin, A.S. , Stahl, S. , Adams, J. , Esseltine, D.‐L. , Elliott, P.J. , Pien, C.S. , Guerciolini, R. , Anderson, J.K. , Depcik‐Smith, N.D. , Bhagat, R. , Lehman, M.J. , Novick, S.C. , O'Connor, O.A. & Soignet, S.L. (2002) Phase I trial of the proteasome inhibitor PS‐341 in patients with refractory hematologic malignancies. Journal of Clinical Oncology, 20, 4420–4427.
- Papadopoulos, K.P. , Burris, H.A. III , Gordon, M. , Lee, P. , Sausville, E.A. , Rosen, P.J. , Patnaik, A. , Cutler, R.E., Jr. , Wang, Z. , Lee, S. , Jones, S.F. & Infante, J.R. (2013) A phase I/II study of carfilzomib 2‐10‐min infusion in patients with advanced solid tumors. Cancer Chemotherapy and Pharmacology, 72, 861–868.
- Papadopoulos, K.P. , Siegel, D.S. , Vesole, D.H. , Lee, P. , Rosen, S.T. , Zojwalla, N. , Holahan, J.R. , Lee, S. , Wang, Z. & Badros, A. (2015) Phase I study of a 30‐minute infusion of carfilzomib as single agent or in combination with low‐dose dexamethasone in patients with relapsed and/or refractory multiple myeloma. Journal of Clinical Oncology, 33, 732–739.
- Papandreou, C.N. , Daliani, D.D. , Nix, D. , Yang, H. , Madden, T. , Wang, X. , Pien, C.S. , Millikan, R.E. , Tu, S.M. , Pagliaro, L. , Kim, J. , Adams, J. , Elliott, P. , Esseltine, D. , Petrusich, A. , Dieringer, P. , Perez, C. & Logothetis, C.J. (2004) Phase I trial of the proteasome inhibitor bortezomib in patients with advanced solid tumors with observations in androgen‐independent prostate cancer. Journal of Clinical Oncology, 22, 108–121.
- Parlati, F. , Lee, S.J. , Aujay, M. , Suzuki, E. , Levitsky, K. , Lorens, J.B. , Micklem, D.R. , Ruurs, P. , Sylvain, C. , Lu, Y. , Shenk, K.D. & Bennett, M.K. (2009) Carfilzomib can induce tumor cell death through selective inhibition of the chymotrypsin‐like activity of the proteasome. Blood, 114, 3439–3447.
- Richardson, P.G. , Sonneveld, P. , Schuster, M.W. , Irwin, D. , Stadtmauer, E.A. , Facon, T. , Harousseau, J.L. , Ben‐Yehuda, D. , Lonial, S. , Goldschmidt, H. , Reece, D. , San‐Miguel, J.F. , Bladé, J. , Boccadoro, M. , Cavenagh, J. , Dalton, W.S. , Boral, A.L. , Esseltine, D.L. , Porter, J.B. , Schenkein, D. & Anderson, K.C. ; for the Assessment of Proteasome Inhibition for Extending Remissions (APEX) Investigators (2005) Bortezomib or high‐dose dexamethasone for relapsed multiple myeloma. New England Journal of Medicine, 352, 2487–2498.
- San‐Miguel, J.F. , Richardson, P.G. , Sonneveld, P. , Schuster, M.W. , Irwin, D. , Stadtmauer, E.A. , Facon, T. , Harousseau, J.L. , Ben‐Yehuda, D. , Lonial, S. , Goldschmidt, H. , Reece, D. , Blade, J. , Boccadoro, M. , Cavenagh, J.D. , Neuwirth, R. , Boral, A.L. , Esseltine, D.L. & Anderson, K.C. (2008) Efficacy and safety of bortezomib in patients with renal impairment: results from the APEX phase 3 study. Leukemia, 22, 842–849.
- Siegel, D.S. , Martin, T. , Wang, M. , Vij, R. , Jakubowiak, A.J. , Lonial, S. , Trudel, S. , Kukreti, V. , Bahlis, N. , Alsina, M. , Chanan‐Khan, A. , Buadi, F. , Reu, F.J. , Somlo, G. , Zonder, J. , Song, K. , Stewart, A.K. , Stadtmauer, E. , Kunkel, L. , Wear, S. , Wong, A.F. , Orlowski, R.Z. & Jagannath, S. (2012) A phase 2 study of single‐agent carfilzomib (PX‐171‐003‐A1) in patients with relapsed and refractory multiple myeloma. Blood, 120, 2817–2825.
- Siegel, D. , Martin, T. , Nooka, A. , Harvey, R.D. , Vij, R. , Niesvizky, R. , Badros, A.Z. , Jagannath, S. , McCulloch, L. , Rajangam, K. & Lonial, S. (2013) Integrated safety profile of single‐agent carfilzomib: experience from 526 patients enrolled in 4 phase 2 clinical studies. Haematologica, 98, 1753–1761.
- Smyth, M.S. & Laidig, G.J. (2006) Compounds for enzyme inhibition (US 2006/0030533 A1), February 9, 2006. . Accessed January 16, 2015.
- Squifflet, P. , Michiels, S. , Siegel, D.S. , Vij, R. , Ro, S. & Buyse, M.E. (2011) Multivariate modelling reveals evidence of a dose‐response relationship in phase 2 studies of single‐agent carfilzomib [abstract]. Blood, 118, Abstract1877.
- Vij, R. , Siegel, D.S. , Jagannath, S. , Jakubowiak, A.J. , Stewart, A.K. , McDonagh, K. , Bahlis, N. , Belch, A. , Kunkel, L.A. , Wear, S. , Wong, A.F. , Orlowski, R.Z. & Wang, W. (2012a) An open‐label, single‐arm, phase 2 study of single‐agent carfilzomib in patients with relapsed and/or refractory multiple myeloma who have been previously treated with bortezomib. British Journal of Haematology, 158, 739–748.
- Vij, R. , Wang, M. , Kaufman, J.L. , Lonial, S. , Jakubowiak, A.J. , Stewart, A.K. , Kukreti, V. , Jagannath, S. , McDonagh, K.T. , Alsina, M. , Bahlis, N.J. , Reu, F.J. , Gabrail, N.Y. , Belch, A. , Matous, J.V. , Lee, P. , Rosen, P. , Sebag, M. , Vesole, D.H. , Kunkel, L.A. , Wear, S.M. , Wong, A.F. , Orlowski, R.Z. & Siegel, D.S. (2012b) An open‐label, single‐arm, phase 2 (PX‐171‐004) study of single‐agent carfilzomib in bortezomib‐naive patients with relapsed and/or refractory multiple myeloma. Blood, 119, 5661–5670.
- Wehenkel, M. , Ban, J.O. , Ho, Y.K. , Carmony, K.C. , Hong, J.T. & Kim, K.B. (2012) A selective inhibitor of the immunoproteasome subunit LMP2 induces apoptosis in PC‐3 cells and suppresses tumour growth in nude mice. British Journal of Cancer, 107, 53–62.
- Wilk, S. & Orlowski, M. (1983) Evidence that pituitary cation‐sensitive neutral endopeptidase is a multicatalytic protease complex. Journal of Neurochemistry, 40, 842–849.
- Yang, J. , Wang, Z. , Fang, Y. , Jiang, J. , Zhao, F. , Wong, H. , Bennett, M.K. , Molineaux, C.J. & Kirk, C.J. (2011) Pharmacokinetics, pharmacodynamics, metabolism, distribution and excretion of carfilzomib in rats. Drug Metabolism and Disposition, 39, 1873–1882.
- Yap, T.A. , Sandhu, S.K. , Workman, P. & de Bono, J.S. (2010) Envisioning the future of early anticancer drug development. Nature Reviews Cancer, 10, 514–523.
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