CeeTox™ Analysis of CNB-001 a Novel Curcumin-Based Neurotrophic/Neuroprotective Lead Compound to Treat Stroke: Comparison with NXY-059 and Radicut
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In the present study, we used a comprehensive cellular toxicity (CeeTox) analysis panel to determine the toxicity profile for CNB-001 [4-((1E)-2-(5-(4-hydroxy-3-methoxystyryl-)-1-phenyl-1H-pyrazoyl-3-yl)vinyl)-2-methoxy-phenol)], which is a hybrid molecule created by combining cyclohexyl bisphenol A, a molecule with neurotrophic activity and curcumin, a spice with neuroprotective activity. CNB-001 is a lead development compound since we have recently shown that CNB-001 has significant preclinical efficacy both in vitro and in vivo. In this study, we compared the CeeTox profile of CNB-001 with two neuroprotective molecules that have been clinically tested for efficacy: the hydrophilic free radical spin trap agent NXY-059 and the hydrophobic free radical scavenger edaravone (Radicut). CeeTox analyses using a rat hepatoma cell line (H4IIE) resulted in estimated C Tox value (i.e., sustained concentration expected to produce toxicity in a rat 14-day repeat dose study) of 42 μM for CNB-001 compared with >300 μM for both NXY-059 and Radicut. The CeeTox panel suggests that CNB-001 produces some adverse effects on cellular adenosine triphosphate content, membrane toxicity, glutathione content, and cell mass (or number), but only with high concentrations of the drug. After a 24-h exposure, the drug concentration that produced a half-maximal response (TC50) on the measures noted above ranges from 55 to 193 μM. Moreover, all CNB-001-induced changes in the markers were coincident with loss of cell number, prior to acute cell death as measured by membrane integrity, suggesting a cytostatic effect of CNB-001. NXY-059 and Radicut did not have acute toxic effects on H4IIE cells. We also found that CNB-001 resulted in an inhibition of ethoxyresorufin-o-deethylase activity, indicating that the drug may affect cytochrome P4501A activity and that CNB-001 was metabolically unstable using a rat microsome assay system. For CNB-001, an estimated in vitro efficacy/toxicity ratio is 183–643-fold, suggesting that there is a significant therapeutic safety window for CNB-001 and that it should be further developed as a novel neuroprotective agent to treat stroke.
- Lapchak PA. Translational stroke research using a rabbit embolic stroke model: a correlative analysis hypothesis for novel therapy development. Translational Stroke Research. 2010;1(2):96–107. doi:10.1007/s12975-010-0018-4 [Perspective Review]. CrossRef
- O'Collins VE, Macleod MR, Donnan GA, Horky LL, van der Worp BH, Howells DW. 1,026 experimental treatments in acute stroke. Ann Neurol. 2006;59(3):467–77. CrossRef
- Feuerstein GZ, Zaleska MM, Krams M, Wang X, Day M, Rutkowski JL, et al. Missing steps in the STAIR case: a Translational Medicine perspective on the development of NXY-059 for treatment of acute ischemic stroke. J Cereb Blood Flow Metab. 2008;28(1):217–9. CrossRef
- Green AR, Ashwood T. Free radical trapping as a therapeutic approach to neuroprotection in stroke: experimental and clinical studies with NXY-059 and free radical scavengers. Curr Drug Targets CNS Neurol Disord. 2005;4(2):109–18. CrossRef
- Durukan A, Tatlisumak T. Acute ischemic stroke: overview of major experimental rodent models, pathophysiology, and therapy of focal cerebral ischemia. Pharmacol Biochem Behav. 2007;87(1):179–97. CrossRef
- Maher P, Salgado KF, Zivin JA, Lapchak PA. A novel approach to screening for new neuroprotective compounds for the treatment of stroke. Brain Res. 2007;1173:117–25. CrossRef
- Liu Y, Dargusch R, Maher P, Schubert D. A broadly neuroprotective derivative of curcumin. J Neurochem. 2008;105(4):1336–45. CrossRef
- Maher P, Akaishi T, Schubert D, Abe K. A pyrazole derivative of curcumin enhances memory. Neurobiol Aging. 2008;31:706–9. CrossRef
- Lapchak PA, Maher P, Schubert D, Zivin JA. Baicalein, an antioxidant 12/15-lipoxygenase inhibitor improves clinical rating scores following multiple infarct embolic strokes. Neuroscience. 2007;150(3):585–91. CrossRef
- Lapchak PA, Schubert D, Maher P. The curcumin derivative CNB-001 attenuates ischemia-induced cell death in vitro and reduces embolism-induced clinical deficits in vivo in a rabbit embolic stroke model: neuroprotection is mediated via an erk-orp150 pathway. Stroke. 2010;e276:85.
- Tan S, Schubert D, Maher P. Oxytosis: A novel form of programmed cell death. Curr Top Med Chem. 2001;1(6):497–506. CrossRef
- Abe K, Takayanagi M, Saito H. Effects of recombinant human basic FGF and its modified protein CS23 on survival of primary cultured neurons from various regions of fetal rat brain. Japan Journal of Pharmacology. 1990;53:221–7. CrossRef
- McKim JM Jr. Building a tiered approach to in vitro predictive toxicity screening: a focus on assays with in vivo relevance. Comb Chem High Throughput Screen. 2010;13(2):188–206. CrossRef
- Bath PM, Gray LJ, Bath AJ, Buchan A, Miyata T, Green AR. Effects of NXY-059 in experimental stroke: an individual animal meta-analysis. Br J Pharmacol. 2009;157(7):1157–71. CrossRef
- Diener HC, Lees KR, Lyden P, Grotta J, Davalos A, Davis SM, et al. NXY-059 for the treatment of acute stroke: pooled analysis of the SAINT I and II Trials. Stroke. 2008;39(6):1751–8. CrossRef
- Shuaib A, Lees KR, Lyden P, Grotta J, Davalos A, Davis SM, et al. NXY-059 for the treatment of acute ischemic stroke. N Engl J Med. 2007;357(6):562–71. CrossRef
- Yoshida H, Yanai H, Namiki Y, Fukatsu-Sasaki K, Furutani N, Tada N. Neuroprotective effects of edaravone: a novel free radical scavenger in cerebrovascular injury. CNS Drug Rev. 2006;12(1):9–20. CrossRef
- Watanabe T, Tahara M, Todo S. The novel antioxidant edaravone: from bench to bedside. Cardiovasc Ther. 2008;26(2):101. CrossRef
- Toyoda K, Fujii K, Kamouchi M, Nakane H, Arihiro S, Okada Y, et al. Free radical scavenger, edaravone, in stroke with internal carotid artery occlusion. J Neurol Sci. 2004;221(1–2):11–7. CrossRef
- Mishina M, Komaba Y, Kobayashi S, Tanaka N, Kominami S, Fukuchi T, et al. Efficacy of edaravone, a free radical scavenger, for the treatment of acute lacunar infarction. Neurol Med Chir (Tokyo). 2005;45(7):344–8. discussion 8. CrossRef
- Inatomi Y, Takita T, Yonehara T, Fujioka S, Hashimoto Y, Hirano T, et al. Efficacy of edaravone in cardioembolic stroke. Intern Med. 2006;45(5):253–7. CrossRef
- Otomo E. Effect of a novel free radical scavenger, edaravone (MCI-186), on acute brain infarction. Randomized, placebo-controlled, double-blind study at multicenters. Cerebrovasc Dis. 2003;15(3):222–9. CrossRef
- Lapchak PA. A critical assessment of edaravone acute ischemic stroke efficacy trials: is edaravone an effective neuroprotective therapy? Expert Opinion on Pharmacotherapy. 2010;11(10):1753–63. doi:10.1517/14656566.2010.493558. CrossRef
- McKim JM, Jr., inventor Ceetox, Inc., assignee. Toxicity screening methods 2009.
- Lapchak PA, Araujo DM, Song D, Wei J, Purdy R, Zivin JA. Effects of the spin trap agent disodium-[tert-butylimino)methyl]benzene-1, 3-disulfonate N-oxide (generic NXY-059) on intracerebral hemorrhage in a rabbit large clot embolic stroke model: combination studies with tissue plasminogen activator. Stroke. 2002;33(6):1665–70. CrossRef
- Lapchak PA, Araujo DM, Song D, Wei J, Zivin JA. Neuroprotective effects of the spin trap agent disodium-[(tert-butylimino)methyl]benzene-1, 3-disulfonate N-oxide (generic NXY-059) in a rabbit small clot embolic stroke model: combination studies with the thrombolytic tissue plasminogen activator. Stroke. 2002;33(5):1411–5. CrossRef
- Lapchak PA, Zivin JA. The lipophilic multifunctional antioxidant edaravone (radicut) improves behavior following embolic strokes in rabbits: a combination therapy study with tissue plasminogen activator. Exp Neurol. 2009;215(1):95–100. CrossRef
- Hopkinson K, Williams EA, Fairburn B, Forster S, Flower DJ, Saxton JM, et al. A MitoTracker Green-based flow cytometric assay for natural killer cell activity: variability, the influence of platelets and a comparison of analytical approaches. Exp Hematol. 2007;35(3):350–7. CrossRef
- Vickers AE. Use of human organ slices to evaluate the biotransformation and drug-induced side-effects of pharmaceuticals. Cell Biol Toxicol. 1994;10(5–6):407–14. CrossRef
- Bernas T, Dobrucki J. Mitochondrial and nonmitochondrial reduction of MTT: interaction of MTT with TMRE, JC-1, and NAO mitochondrial fluorescent probes. Cytometry. 2002;47(4):236–42. CrossRef
- Berridge MV, Tan AS. Characterization of the cellular reduction of 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT): subcellular localization, substrate dependence, and involvement of mitochondrial electron transport in MTT reduction. Arch Biochem Biophys. 1993;303(2):474–82. CrossRef
- Lapchak PA, De Taboada L. Transcranial near infrared laser treatment (NILT) increases cortical adenosine-5'-triphosphate (ATP) content following embolic strokes in rabbits. Brain Res. 2010;1306:100–5. CrossRef
- Griffith OW. Determination of glutathione and glutathionedisulfide using glutathione reductase and 2-vinylpyridine. Anal Biochem. 1980;106:207–12. CrossRef
- Vacchiano CA, Tempel GE. Role of nonenzymatically generated prostanoid, 8-iso-PGF2 alpha, in pulmonary oxygen toxicity. J Appl Physiol. 1994;77(6):2912–7.
- Muller H, Klinkhammer W, Globisch C, Kassack MU, Pajeva IK, Wiese M. New functional assay of P-glycoprotein activity using Hoechst 33342. Bioorg Med Chem. 2007;15(23):7470–9. CrossRef
- Gao GL, Wan HY, Zou XS, Chen WX, Chen YQ, Huang XZ. Relationship between the expression of P-glycoprotein, glutathione S-transferase-pi and thymidylate synthase proteins and adenosine triphosphate tumor chemosensitivity assay in cervical cancer. Zhonghua Fu Chan Ke Za Zhi. 2007;42(3):201–5.
- Li QY, Wang Y, Yin ZF, Wu MC. Application of the improved MTT assay in predicting the intrinsic drug resistance of liver cancer. Zhonghua Yi Xue Za Zhi. 2007;87(5):333–5.
- Burke MD, Thompson S, Elcombe CR, Halpert J, Haaparanta T, Mayer RT. Ethoxy-, pentoxy- and benzyloxyphenoxazones and homologues: a series of substrates to distinguish between different induced cytochromes P-450. Biochem Pharmacol. 1985;34(18):3337–45. CrossRef
- D'Amelio M, Cavallucci V, Cecconi F. Neuronal caspase-3 signaling: not only cell death. Cell Death Differ. 2009;17:1104–14. doi:10.1038/cdd.2009.180. CrossRef
- McKim JM, Jr., inventor Ceetox, Inc., assignee. Toxicity screening methods 2007.
- Green AR. Why do neuroprotective drugs that are so promising in animals fail in the clinic? An industry perspective. Clin Exp Pharmacol Physiol. 2002;29(11):1030–4. CrossRef
- Legos JJ, Tuma RF, Barone FC. Pharmacological interventions for stroke: failures and future. Expert Opin Investig Drugs. 2002;11(5):603–14. CrossRef
- Liebeskind DS, Kasner SE. Neuroprotection for ischaemic stroke: an unattainable goal? CNS Drugs. 2001;15(3):165–74. CrossRef
- Higashi Y, Jitsuiki D, Chayama K, Yoshizumi M. Edaravone (3-methyl-1-phenyl-2-pyrazolin-5-one), a novel free radical scavenger, for treatment of cardiovascular diseases. Recent Patents Cardiovasc Drug Discov. 2006;1(1):85–93. CrossRef
- Lapchak PA. NXY-059. Centaur Curr Opin Investig Drugs. 2002;3(12):1758–62.
- Lapchak PA, Araujo DM. Development of the Nitrone-Based Spin Trap Agent NXY-059 to treat acute ischemic stroke. CNS Drug Rev. 2003;9(3):253–62. CrossRef
- Maples KR, Green AR, Floyd RA. Nitrone-related therapeutics: potential of NXY-059 for the treatment of acute ischaemic stroke. CNS Drugs. 2004;18(15):1071–84. CrossRef
- Maples KR, Ma F, Zhang YK. Comparison of the radical trapping ability of PBN, S-PPBN and NXY-059. Free Radic Res. 2001;34(4):417–26. CrossRef
- Hishida A. Clinical analysis of 207 patients who developed renal disorders during or after treatment with edaravone reported during post-marketing surveillance. Clin Exp Nephrol. 2007;11(4):292–6. CrossRef
- FDA. Guidance for Industry M4S: The CTD—Safety Appendices. 2001. http://www.fda.gov/cder/guidance/index.htm.
- CeeTox™ Analysis of CNB-001 a Novel Curcumin-Based Neurotrophic/Neuroprotective Lead Compound to Treat Stroke: Comparison with NXY-059 and Radicut
Translational Stroke Research
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