PAC-1

PAC-1 was discovered in Professor Paul Hergenrother's laboratory at the University of Illinois at Urbana–Champaign during a process that screened many chemicals for anti-tumor potential. This molecule, when delivered to cancer cells, signals the cells to self-destruct by activating an "executioner" protein, procaspase-3. Then, the activated executioner protein begins a cascade of events that destroys the machinery of the cell. In 2011, Vanquish Oncology Inc. was founded to move PAC-1 forward to a human clinical trial. In 2013, Vanquish announced a multimillion-dollar angel investment into the company. In 2015, a phase I clinical trial of PAC-1 opened for enrollment of cancer patients, and in 2016, it was announced that PAC-1 had been granted Orphan Drug Designation for treatment of glioblastoma by the FDA, and in late 2017 a Phase 1b trial began of PAC-1 plus temozolomide for treatment of patients with recurrent glioblastoma or anaplastic astrocytoma.{{cn|date=February 2023}}

In cells, the executioner protein, caspase-3, is stored in its inactive form, procaspase-3. This way, the cell can quickly undergo apoptosis by activating the protein that is already there. This inactive form is called a zymogen. Procaspase-3 is known to be inhibited by low levels of zinc. PAC-1 activates procaspase-3 by chelating zinc, thus relieving the zinc-mediated inhibition. This allows procaspase-3 to be an active enzyme, and it can then cleave another molecule of procaspase-3 to active caspase-3. Caspase-3 can further activate other molecules of procaspase-3 in the cell, causing an exponential increase in caspase-3 concentration. PAC-1 facilitates this process and causes the cell to undergo apoptosis quickly.{{cite journal | vauthors = Putt KS, Chen GW, Pearson JM, Sandhorst JS, Hoagland MS, Kwon JT, Hwang SK, Jin H, Churchwell MI, Cho MH, Doerge DR, Helferich WG, Hergenrother PJ | display-authors = 6 | title = Small-molecule activation of procaspase-3 to caspase-3 as a personalized anticancer strategy | journal = Nature Chemical Biology | volume = 2 | issue = 10 | pages = 543–550 | date = October 2006 | pmid = 16936720 | doi = 10.1038/nchembio814 | s2cid = 1689173 }}

This direct procaspase-3 activation mode-of-action for PAC-1 has been confirmed by other laboratories: In 2013 by the Megeney lab during the course of studies on the role of caspase-3 in cardiomyocytes,{{cite journal | vauthors = Putinski C, Abdul-Ghani M, Stiles R, Brunette S, Dick SA, Fernando P, Megeney LA | title = Intrinsic-mediated caspase activation is essential for cardiomyocyte hypertrophy | journal = Proceedings of the National Academy of Sciences of the United States of America | volume = 110 | issue = 43 | pages = E4079–E4087 | date = October 2013 | pmid = 24101493 | pmc = 3808644 | doi = 10.1073/pnas.1315587110 | bibcode = 2013PNAS..110E4079P | doi-access = free }}

in 2014 by the Wu lab in an extensive study on the anticancer activity and mode-of-action of PAC-1 and derivatives,{{cite journal | vauthors = Wang F, Wang L, Zhao Y, Li Y, Ping G, Xiao S, Chen K, Zhu W, Gong P, Yang J, Wu C | display-authors = 6 | title = A novel small-molecule activator of procaspase-3 induces apoptosis in cancer cells and reduces tumor growth in human breast, liver and gallbladder cancer xenografts | journal = Molecular Oncology | volume = 8 | issue = 8 | pages = 1640–1652 | date = December 2014 | pmid = 25053517 | pmc = 5528581 | doi = 10.1016/j.molonc.2014.06.015 }} and in 2015 by the Gandhi lab in an exploration of the potential of PAC-1 and derivative B-PAC-1 for treatment of chronic lymphocytic leukemia (CLL).{{cite journal | vauthors = Patel V, Balakrishnan K, Keating MJ, Wierda WG, Gandhi V | title = Expression of executioner procaspases and their activation by a procaspase-activating compound in chronic lymphocytic leukemia cells | journal = Blood | volume = 125 | issue = 7 | pages = 1126–1136 | date = February 2015 | pmid = 25538042 | pmc = 4326772 | doi = 10.1182/blood-2014-01-546796 }}

Studies with knockout cells have suggested the importance of procaspase-7 as a secondary or alternate target for PAC-1, especially in the absence of procaspase-3. For example, experiments using mouse embryonic fibroblasts (MEFs) demonstrate that the double knockout of the CASP3 and CASP7 genes leads to cells that are insensitive to the proapoptotic effects of the PAC-1 class of compounds, and knocking in either CASP3 or CASP7 once again sensitizes these cells to PAC-1-type compounds.{{cite journal | vauthors = Sarkar A, Balakrishnan K, Chen J, Patel V, Neelapu SS, McMurray JS, Gandhi V | title = Molecular evidence of Zn chelation of the procaspase activating compound B-PAC-1 in B cell lymphoma | journal = Oncotarget | volume = 7 | issue = 3 | pages = 3461–3476 | date = January 2016 | pmid = 26658105 | pmc = 4823120 | doi = 10.18632/oncotarget.6505 }}

Recent experiments using cancer cell lines with CRISPR deletion of CASP3 are also consistent with this result.{{cite journal | vauthors = Lin A, Giuliano CJ, Palladino A, John KM, Abramowicz C, Yuan ML, Sausville EL, Lukow DA, Liu L, Chait AR, Galluzzo ZC, Tucker C, Sheltzer JM | display-authors = 6 | title = Off-target toxicity is a common mechanism of action of cancer drugs undergoing clinical trials | journal = Science Translational Medicine | volume = 11 | issue = 509 | pages = eaaw8412 | date = September 2019 | pmid = 31511426 | pmc = 7717492 | doi = 10.1126/scitranslmed.aaw8412 }} The activation of procaspase-7 by PAC-1 is consistent with biochemical data, although the relative importance of the procaspase-7 target in cells with functional procaspase-3 is uncertain.{{cn|date=February 2023}}

A potential selectivity problem arises because procaspase-3 is present in most cells of the body. However, it has been shown that in many cancers, including certain neuroblastomas, lymphomas, leukemias, melanomas, and liver cancers, procaspase-3 is present in higher concentrations. For instance, lung cancer cells can have over 1000 times more procaspase-3 than normal cells. Therefore, by controlling the dosage, one can achieve selectivity between normal and cancerous cells. {{cn|date=November 2024}}

In addition to its stand-alone activity, PAC-1 has also been shown to markedly synergize with a variety of approved cancer drugs, for example with BRAF and MEK inhibitors in mouse models of melanoma,{{cite journal | vauthors = Peh J, Fan TM, Wycislo KL, Roth HS, Hergenrother PJ | title = The Combination of Vemurafenib and Procaspase-3 Activation Is Synergistic in Mutant BRAF Melanomas | journal = Molecular Cancer Therapeutics | volume = 15 | issue = 8 | pages = 1859–1869 | date = August 2016 | pmid = 27297867 | pmc = 4975653 | doi = 10.1158/1535-7163.MCT-16-0025 }}

and with conventional chemotherapeutic agents such as doxorubicin in pet dogs with spontaneous cancers including lymphoma and metastatic osteosarcoma,{{cite journal | vauthors = Botham RC, Roth HS, Book AP, Roady PJ, Fan TM, Hergenrother PJ | title = Small-Molecule Procaspase-3 Activation Sensitizes Cancer to Treatment with Diverse Chemotherapeutics | journal = ACS Central Science | volume = 2 | issue = 8 | pages = 545–559 | date = August 2016 | pmid = 27610416 | pmc = 4999974 | doi = 10.1021/acscentsci.6b00165 }} and with temozolomide in pet dogs with naturally occurring glioma.{{cite journal | vauthors = Joshi AD, Botham RC, Schlein LJ, Roth HS, Mangraviti A, Borodovsky A, Tyler B, Joslyn S, Looper JS, Podell M, Fan TM, Hergenrother PJ, Riggins GJ | display-authors = 6 | title = Synergistic and targeted therapy with a procaspase-3 activator and temozolomide extends survival in glioma rodent models and is feasible for the treatment of canine malignant glioma patients | journal = Oncotarget | volume = 8 | issue = 46 | pages = 80124–80138 | date = October 2017 | pmid = 29113289 | pmc = 5655184 | doi = 10.18632/oncotarget.19085 }}

Vanquish Oncology reported their intention to begin a Phase I human clinical trial in cancer patients to begin in early 2015, and indeed the Phase 1 trial of PAC-1 opened for enrollment in February 2015 (NCT02355535). This trial is being conducted at the University of Illinois Cancer Center in Chicago, at the Sidney Kimmel Cancer Center at Johns Hopkins, and at Regions Hospital in St. Paul, MN. A Phase 1b trial of PAC-1 plus temozolomide opened in late 2017 at the same three sites (NCT03332355); patients with high grade glioma (glioblastoma multiforme (GBM) or anaplastic astrocytoma) after progression following standard first line therapy are eligible for this trial. {{cn|date=November 2024}}

PAC-1 is notable for the unique path it has taken to the clinic, as it is possibly the only cancer drug to first be rigorously evaluated in pet dogs with spontaneous cancer as a prelude to the human clinical trial. In 2010, a study showed PAC-1 to be safe for dogs, and a second study published later that same year reported that a PAC-1 derivative (called S-PAC-1) was well tolerated in a small phase I clinical trial of dogs with lymphoma. More recently, in addition to this single-agent activity PAC-1 has shown to potently synergize with approved cancer drugs, for example with doxorubicin in the treatment of pet dogs with lymphoma and metastatic osteosarcoma, and with temozolomide in treating pet dogs with spontaneous glioma.

PAC-1 has been or is currently being tested in the following human clinical trials:

1. {{ClinicalTrialsGov|NCT02355535|Procaspase Activating Compound-1 (PAC-1) in the Treatment of Advanced Malignancies - Component 1}}
2. {{ClinicalTrialsGov|NCT03332355|Procaspase Activating Compound-1 (PAC-1) in the Treatment of Advanced Malignancies - Component 2}}
3. {{ClinicalTrialsGov|NCT03927248|PAC-1 for Treatment of Refractory, Metastatic Kidney Cancer}}

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• {{cite journal | vauthors = Peterson QP, Goode DR, West DC, Ramsey KN, Lee JJ, Hergenrother PJ | title = PAC-1 activates procaspase-3 in vitro through relief of zinc-mediated inhibition | journal = Journal of Molecular Biology | volume = 388 | issue = 1 | pages = 144–158 | date = April 2009 | pmid = 19281821 | pmc = 2714579 | doi = 10.1016/j.jmb.2009.03.003 }}
• {{cite journal | vauthors = Peterson QP, Hsu DC, Goode DR, Novotny CJ, Totten RK, Hergenrother PJ | title = Procaspase-3 activation as an anti-cancer strategy: structure-activity relationship of procaspase-activating compound 1 (PAC-1) and its cellular co-localization with caspase-3 | journal = Journal of Medicinal Chemistry | volume = 52 | issue = 18 | pages = 5721–5731 | date = September 2009 | pmid = 19708658 | pmc = 2749958 | doi = 10.1021/jm900722z }}
• {{cite journal | vauthors = Lucas PW, Schmit JM, Peterson QP, West DC, Hsu DC, Novotny CJ, Dirikolu L, Churchwell MI, Doerge DR, Garrett LD, Hergenrother PJ, Fan TM | display-authors = 6 | title = Pharmacokinetics and derivation of an anticancer dosing regimen for PAC-1, a preferential small molecule activator of procaspase-3, in healthy dogs | journal = Investigational New Drugs | volume = 29 | issue = 5 | pages = 901–911 | date = October 2011 | pmid = 20499133 | pmc = 3182491 | doi = 10.1007/s10637-010-9445-z }}
• {{cite journal | vauthors = Peterson QP, Hsu DC, Novotny CJ, West DC, Kim D, Schmit JM, Dirikolu L, Hergenrother PJ, Fan TM | display-authors = 6 | title = Discovery and canine preclinical assessment of a nontoxic procaspase-3-activating compound | journal = Cancer Research | volume = 70 | issue = 18 | pages = 7232–7241 | date = September 2010 | pmid = 20823163 | pmc = 3113694 | doi = 10.1158/0008-5472.can-10-0766 }}
• {{cite journal | vauthors = West DC, Qin Y, Peterson QP, Thomas DL, Palchaudhuri R, Morrison KC, Lucas PW, Palmer AE, Fan TM, Hergenrother PJ | display-authors = 6 | title = Differential effects of procaspase-3 activating compounds in the induction of cancer cell death | journal = Molecular Pharmaceutics | volume = 9 | issue = 5 | pages = 1425–1434 | date = May 2012 | pmid = 22486564 | pmc = 3348238 | doi = 10.1021/mp200673n }}
• {{cite journal | vauthors = Botham RC, Fan TM, Im I, Borst LB, Dirikolu L, Hergenrother PJ | title = Dual small-molecule targeting of procaspase-3 dramatically enhances zymogen activation and anticancer activity | journal = Journal of the American Chemical Society | volume = 136 | issue = 4 | pages = 1312–1319 | date = January 2014 | pmid = 24383395 | pmc = 3954530 | doi = 10.1021/ja4124303 }}

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• [http://news.bbc.co.uk/2/hi/health/5284850.stm Cancer cell 'executioner' found]. BBC News 27 August 2006.
• [http://news.bbc.co.uk/2/hi/health/3670495.stm Cancer cells 'can live forever']. BBC News 29 April 2004.
• [http://vanquishoncology.com Vanquish Oncology]
• [http://news.illinois.edu/news/13/0717cancer_drug_PaulHergenrother_TimFan.html Angel investment helps move PAC-1 forward]
• [https://news.illinois.edu/blog/view/6367/365713 FDA grants PAC-1 Orphan Drug Designation for glioma]
• [https://clinicaltrials.gov/ct2/show/NCT02355535 Phase 1 clinical trial of PAC-1 in human cancer patients] clinicaltrials.gov

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Category:Cell signaling

Category:Experimental cancer drugs

Category:Immune system

Category:Programmed cell death