Identification of Small Molecule Inhibitors of Human Cytochrome c Oxidase That Target Chemoresistant Glioma Cells.
The enzyme cytochrome c oxidase (CcO) or complex IV (EC 1.9.3.1) is a large transmembrane protein complex that serves as the last enzyme in the respiratory electron transport chain of eukaryotic mitochondria. CcO promotes the switch from glycolytic to oxidative phosphorylation (OXPHOS) metabolism and has been associated with increased self-renewal characteristics in gliomas. Increased CcO activity in tumors has been associated with tumor progression after chemotherapy failure, and patients with primary glioblastoma multiforme and high tumor CcO activity have worse clinical outcomes than those with low tumor CcO activity. Therefore, CcO is an attractive target for cancer therapy. We report here the characterization of a CcO inhibitor (ADDA 5) that was identified using a high throughput screening paradigm. ADDA 5 demonstrated specificity for CcO, with no inhibition of other mitochondrial complexes or other relevant enzymes, and biochemical characterization showed that this compound is a non-competitive inhibitor of cytochrome c When tested in cellular assays, ADDA 5 dose-dependently inhibited the proliferation of chemosensitive and chemoresistant glioma cells but did not display toxicity against non-cancer cells. Furthermore, treatment with ADDA 5 led to significant inhibition of tumor growth in flank xenograft mouse models. Importantly, ADDA 5 inhibited CcO activity and blocked cell proliferation and neurosphere formation in cultures of glioma stem cells, the cells implicated in tumor recurrence and resistance to therapy in patients with glioblastoma. In summary, we have identified ADDA 5 as a lead CcO inhibitor for further optimization as a novel approach for the treatment of glioblastoma and related cancers.
From the Department of Neurosurgery.
Claudia R
CR
Oliva
From the Department of Neurosurgery.
Tahireh
T
Markert
Drug Discovery Division, Southern Research, Birmingham, Alabama 35205.
Larry J
LJ
Ross
Drug Discovery Division, Southern Research, Birmingham, Alabama 35205.
E Lucile
EL
White
Drug Discovery Division, Southern Research, Birmingham, Alabama 35205.
Lynn
L
Rasmussen
Drug Discovery Division, Southern Research, Birmingham, Alabama 35205.
Wei
W
Zhang
Department of Pediatrics, Division of Infectious Diseases, University of Alabama at Birmingham, Birmingham, Alabama 35294.
Maaike
M
Everts
UAB Nutrition Sciences Department, Diabetes Research Center BARB Core, University of Alabama at Birmingham, Birmingham, Alabama 35294.
Douglas R
DR
Moellering
Department of Pathology, Division of Molecular and Cellular Pathology, University of Alabama at Birmingham, Birmingham, Alabama 35294, and.
Shannon M
SM
Bailey
Drug Discovery Division, Southern Research, Birmingham, Alabama 35205.
Mark J
MJ
Suto
From the Department of Neurosurgery, cegriguer@uabmc.edu.|Center for Free Radical Biology, and.
Corinne E
CE
Griguer
D000818:Animals; D045744:Cell Line, Tumor; D045304:Cytochromes c / Q000378:metabolism; D019008:Drug Resistance, Neoplasm / Q000187:drug effects*; D003576:Electron Transport Complex IV / Q000037:antagonists & inhibitors* / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005910:Glioma / Q000188:drug therapy / Q000201:enzymology; D006801:Humans; D051379:Mice; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D023041:Xenograft Model Antitumor Assays
D004791:Enzyme Inhibitors; D009363:Neoplasm Proteins; D045304:Cytochromes c; D003576:Electron Transport Complex IV
chemoresistance; cytochrome c oxidase (complex IV); glioblastoma; inhibitor; metabolism; mitochondria; small molecule; stem cells
Cancer Res. 1980 Dec;40(12):4677-81
6893680
Free Radic Biol Med. 2010 Apr 1;48(7):905-14
20093177
Arch Biochem Biophys. 1964 Sep;107:537-43
14234506
J Biol Chem. 1998 Nov 27;273(48):31829-36
9822650
Anal Biochem. 1996 Jun 1;237(2):274-8
8660576
Proc Natl Acad Sci U S A. 1989 Mar;86(5):1583-7
2922400
Cancer Res. 2007 Jun 1;67(11):5239-47
17545603
J Exp Biol. 1998 Apr;201(Pt 8):1129-39
9510525
J Neurooncol. 2005 Sep;74(2):123-33
16193382
Annu Rev Pharmacol Toxicol. 1984;24:451-81
6428300
Nat Protoc. 2010 Jan;5(1):51-66
20057381
J Biomol Screen. 1999;4(2):67-73
10838414
PLoS One. 2008;3(11):e3655
18985161
J Neurooncol. 2011 Jan;101(2):179-88
20532954
Chem Biol Interact. 2011 May 30;191(1-3):288-95
21147079
J Biol Chem. 1981 Oct 25;256(20):10319-28
6270107
Biochem J. 2002 Apr 15;363(Pt 2):321-7
11931660
Redox Biol. 2013;1(1):86-93
23667828
Lab Invest. 1981 Jan;44(1):13-7
6256583
Biochem J. 1984 May 15;220(1):57-66
6331413
Curr Protoc Hum Genet. 2009 Oct;Chapter 19:Unit19.3
19806590
Cancer Res. 2006 Feb 15;66(4):2257-63
16489029
Biochem J. 1994 Aug 15;302 ( Pt 1):171-4
8068003
Oncotarget. 2015 Feb 28;6(6):4330-44
25726526
Cancers (Basel). 2010 Feb 12;2(1):68-87
20532186
PLoS One. 2013 Apr 10;8(4):e61035
23593382
Ciba Found Symp. 1988;140:232-43
3073059
Anal Biochem. 1969 Mar;27(3):502-22
4388022
Acad Radiol. 2013 Jul;20(7):795-7
23747151
J Pharmacol Exp Ther. 1998 Apr;285(1):236-41
9536016
J Neurosci Methods. 2013 Apr 15;214(2):156-62
23376420
J Biol Chem. 2010 Dec 17;285(51):39759-67
20870728
J Biol Chem. 2003 Sep 19;278(38):36027-31
12840017
J Biol Chem. 2004 Nov 5;279(45):46580-7
15328348
Br J Cancer. 2016 Jun 14;114(12):1305-12
27219018
Anal Biochem. 1989 Nov 15;183(1):195-6
2619044
PLoS One. 2011;6(9):e24665
21931801
Curr Neurol Neurosci Rep. 2005 May;5(3):198-206
15865885
N Engl J Med. 2005 Mar 10;352(10 ):987-96
15758009