Doxorubicin

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Doxorubicin
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Executive Summary Information

Compound Doxorubicin
Toxicities Cytotoxicity
Mechanisms Toxicity is initiated by oxidative damage associated both with the hydroquinone moiety and with iron-complexes of the parent compound. The major metabolic product is more toxic than the parent, but metabolism is not a requirement for toxicity. This compound intercalates with DNA and thus causes direct damage to DNA as well as to proteins. Toxicity is both acute and chronic and is life-threatening.
Comments This compound was selected as an archetypical repeated dose cardiotoxin.
Feedback Contact Gold Compound Working Group (GCWG)
Doxorubicin
Doxorubicin.png
Identifiers
Leadscope Id LS-1029
CAS 23214-92-8
DrugBank DB00997
ChemSpider 29400
UNII 80168379AG
ChEBI 28748
Pathway DBs
KEGG D03899
Assay DBs
PubChem CID 31703
ChEMBL 179
Omics DBs
Open TG-Gate 00149
Properties
ToxCast Accepted yes
ToxBank Accepted yes
Approved on 2012-01-01
Target DNA oxidation
Toxicities Cytotoxicity


In Vivo Data ? Compound Assessment
Adverse Events ? Acute cardiotoxicity

Arrythmias during or within 24 hours of doxorubicin administration. Histopathological features of acute cardiotoxicity include increased hyaline material, contraction band necrosis and an infiltrate of neutrophils, lymphocytes and histiocytes.

Subacute cardiotoxicity

Myopericarditis days to weeks after administration.

Chronic cardiotoxicity

The incidence of toxicity is very high: 5% at 400 mg/m2 and 26% at 550 mg/m2. Late onset effects up to 20 years after completion of anthracycline therapy. Dose-dependent progressive myocardial dysfunction. The cellular lesions are known as "myofibrillar dropout", which consist of swelling of the sarcoplasmic reticulum increasing to complete loss of myofibrils.

Several mechanisms for the doxorubicin-induced cardio toxicity have been proposed, including membrane lipid peroxidation, free radical formation, mitochondrial damage and iron-dependent oxidative damage. Oxygen radicals are apparently involved in all mechanisms proposed.

At low concentrations, (P53-mediated) apoptosis is observed, whereas at higher concentrations necrosis is observed. A number of pathways have been proposed for translating oxidative damage to cytotoxicity: an irreversible decrease in mitochondrial calcium loading capacity; modification of cardiolipin; induction of endothelin; deregulation of nitric oxide; and toxicity via reactive metabolites. The relative contribution of these separate mechanisms is not clear quantitatively.

Involvement of an immunogenic reaction after oxidative stress is also implicated in some cases.

References:

-G. J. Berry and M. Jorden ,"Review: Pathology of Radiation and Anthracycline Cardiotoxicity", Pediatr Blood Cancer 2005;44:630–637
-B. N. Bernaba et al. "Pathology of late-onset anthracycline cardiomyopathy" Cardiovascular Pathology 19 (2010) 308–311
-Emanuel Raschi, Valentina Vasina, Maria Grazia Ursino, Giuseppe Boriani, Andrea Martoni, Fabrizio De Ponti, “Anticancer drugs and cardiotoxicity: Insights and perspectives in the era of targeted therapy”, Pharmacology & Therapeutics 125 (2010) 196–218.
Toxicity Mechanisms ? The underlying mechanism of doxorubicin cardiomyopathy is oxidative damage. The effects of this oxidative reactivity include direct DNA damage, accumulation of mitochondrial DNA mutations, alterations in calcium handling, proteolysis of titin, and dysregulation of cardiac transcription factors. Damage is selective but not exclusive for DNA because of intercalation of doxorubicin.

Oxidative reactivity is generated via hydroquinone-quinone redox cycling from the hydroquinone moiety of the parent drug and via complexation of the drug with iron. The relative importance of these two pathways is not fully established.

References:

-M. S. Ewer and St. M. Ewer "Cardiotoxicity of Anticancer Treatments: What the Cardiologist Needs to Know" , Nature Reviews Cardiology 7, 564-575 (October 2010)
Therapeutic Target ? Doxorubicin is an antineoplastic in the anthracycline class. General properties include: interaction with DNA in a variety of different ways including intercalation, DNA strand breakage and inhibition with the enzyme topoisomerase II.

References:

-Reference: http://www.drugbank.ca/drugs/DB00997

FDA and Label Information

The following link will display all of the currently approved FDA drug products on the market. The web page will contain a table listing all current products by their respective Tradenames and primary active ingredients. The list is navigable by simply clicking on the product of interest, which will in turn list all of the NDA's and ANDA's associated with that product. From here users can click on a specific NDA or ANDA and see documents that have been submitted for the product that the FDA has made available via their website. The types of documents include approval history, letters, reviews and labels.
FDA Approved Products

This next url will perform a search in the FDA's system for all labels for products that contain Doxorubicin as an active ingredient.
FDA Label Search

Since every unique drug product receives a separate label, we have included a link to another resource for a specific Doxorubicin product (Doxorubicin hydrochloride injection, solution by Pfizer Laboratories Div Pfizer Inc.) which displays the drug label information.
label

PubMed references

The following resource link will perform a PubMed query for the terms "doxorubicin" and "cardio toxicity".
Doxorubicin Search

PK-ADME ? Compound Assessment
PK parameters ? The most commonly used dose schedule when used as a single agent is 60 to 75 mg/m2 as a single intravenous injection administered at 21-day intervals.
Protein binding70%
Half life55 hours
Vd20-30 L/kg (700-100 L/m2)
Cmax3 uM for 30 mg/m2 intravenous bolus dose. Cellular levels are about 30–100-fold higher than that of the plasma.
Excretionpredominantly in bile, 40-50% in feces within 7 days (50% as unchanged drug).
Plasma clearance324 to 809 mL/min/m2, biphasic
Metabolism~50% metabolized by the liver

References:

-http://www.drugbank.ca/drugs/DB00997
-http://reference.medscape.com/drug/doxorubicin-342120
-A.-K. Souid et al. "Immediate effects of anticancer drugs on mitochondrial oxygen consumption"; Biochemical Pharmacology 66 (2003) 977–987
Therapeutic window ?
Metabolically activated ? There are 2 main metabolic routes of anthracycline metabolism: two-electron reduction and deglycosidation. A large proportion of Doxorubicin (DOX) however, approximately 50%, is eliminated from the body unchanged.

Two-electron reduction of DOX to a secondary alcohol, Doxorubicinol (DOXol, up to 10 times more potent than DOX) is the major metabolic pathway. Several enzymes can carry out this reaction and their respective balance is different in different cell types. AKR1A is considered the most important in heart tissue while CBR1 is the major contributor in liver.

Deglycosidation, the third, minor route, accounts for approximately 1-2% of DOX metabolism. This can be reductive to form the deoxyaglycone, or hydrolytic to form the hydroxyaglycone. The enzymes and their candidate genes for this process are less well characterized. In heart, no DOX hydroxyaglycone could be detected.

DOX metabolism in human myocardium:

Human myocardium convert DOX to DOXol by virtue of different reductases. Besides DOXol myocardium can generate DOX deoxyaglycone and DOXol hydroxyaglycone, reflecting reduction of the side chain carbonyl group, reductase-type deglycosidation of the anthracycline, and hydrolase-type deglycosidation followed by carbonyl reduction, respectively.

Intramyocardial formation of secondary alcohol metabolites might play a key role in promoting the progression of cardiotoxicity.

References:

-Mordente A et al.,"New developments in anthracycline-induced cardiotoxicity" Curr Med Chem. 2009;16(13):1656-72.
-Kassner N. et al. Carbonyl reductase 1 is a predominant doxorubicin reductase in the human liver" Drug Metab Dispos. 2008 Oct;36(10):2113-20. Epub 2008 Jul 17.
-S. Licata et al.; "Doxorubicin Metabolism and Toxicity in Human Myocardium:  Role of Cytoplasmic Deglycosidation and Carbonyl Reduction"; Chem. Res. Toxicol., 2000, 13 (5), pp 414–420
-Thorn Caroline F et al. "Doxorubicin pathways: pharmacodynamics and adverse effects" Pharmacogenetics and genomics (2010)
-A. Mordente et al. "Anthracycline secondary alcohol metabolite formation in human or rabbit heart: biochemical aspects and pharmacologic implications" Biochemical Pharmacology 66 (2003) 989–998

Omics and IC50 Data ? Compound Assessment
Gene expression profiles known. ? References:
-Amy V. Pointon, Tracy M. Walker, Kate M. Phillips, Jinli Luo, Joan Riley, Shu-Dong Zhang, Joel D. Parry, Jonathan J. Lyon, Emma L. Marczylo, Timothy W. Gant, “Doxorubicin In Vivo Rapidly Alters Expression and Translation of Myocardial Electron Transport Chain Genes, Leads to ATP Loss and Caspase 3 Activation”, (2010) PLoS ONE 5(9): e12733. doi:10.1371/journal.pone.0012733.
-Raju Jeyaseelan, Coralie Poizat, Robert K. Bakeri, Serge Abdishoo, Larissa B. Isterabadi, Gary E. Lyonsi, and Larry Kedes, ” A Novel Cardiac-Restricted Target for Doxorubicin”, J. Biol. Chem. Vol. 272:22800–22808, 1997.
-M. Tokarska-Schlattner et al. "Earlyeffects of doxorubicin in perfused heart: transcriptional profiling revealsinhibition of cellular stress response genes" AJP - Regu Physiol April 2010 vol. 298 no. 4
-Pointon AV et al. "Doxorubicin In Vivo Rapidly Alters Expression and Translation of Myocardial Electron Transport Chain Genes, Leads to ATP Loss and Caspase 3 Activation".(2010) PLoS ONE 5(9): e12733

ABCT gene expression in doxorubicin treated HepG2 cells

-Jodi Morrison et al. "DTExtm - Gene Expression Profiling using a Human ABC Transporter Microarray"
-Seongeun Lee et al. "Mechanism of doxorubicin-induced cell death and expression profile analysis" Biotechnology Letters 24: 1147–1151, 2002
-Maria A. Folgueira et al."Gene Expression Profile Associated with Response to Doxorubicin-Based Therapy in Breast Cancer" Clin Cancer Res October 15, 2005 11; 7434
Proteomics profiles known. ? References:
-Kumar SN et al., "Analysis of proteome changes in doxorubicin-treated adult rat cardiomyocyte" J Proteomics. 2011 May 1;74(5):683-97.
Metabonomics profiles known. ? References:
-A. Strigun et al. "Metabolic profiling using HPLC allows classification of drugs according to their mechanisms of action in HL-1 cardiomyocytes" Toxicology and Applied Pharmacology, Volume 252, Issue 2, 15 April 2011, Pages 183-191
Fluxomics profiles known. ?
Epigenomics profiles known. ? Characterization of DNA methylation changes which arise from treatment of tumor cells with the chemotherapeutic drug doxorubicin.

References:

-Boettcher M et al., "High-Definition DNA Methylation Profiles from Breast and Ovarian Carcinoma Cell Lines with Differing Doxorubicin Resistance". PLoS ONE (2010) 5(6): e11002

Alterations in epigenetic mechanisms in the drug-resistant MCF-7 human breast cancer cells induced by doxorubicin.

References:

-Vasyl' F. Chekhun et al,"Epigenetic profiling of multidrug-resistant human MCF-7 breast adenocarcinoma cells reveals novel hyper- and hypomethylated targets" Mol Cancer Ther March 2007 6; 1089
Observed IC50 for in vitro cellular efficacy. ? Human breast tumor cell lines, IC50, uM
HTB-1230.056
HTB-261.35
MCF-70.6
HTB-190.83
HTB-201.10

References:

-Yoonik Lee et al. "Effects on non-cytotoxic concentration of anticancer drugs on doxorubicin cytotoxicity in human breast cancer cell lines" J.Biochem. Mol. Biol. Vol 29, No 4, pp 314-320
Observed IC50 for in vitro cellular toxicity studies. ? Rat LD50 = 21.8 mg/kg (40 umol/kg, subcutaneous)
In-vitro rat myocardiocytes at 24h1.23 μg/mL (2 uM)
In-vitro rat myocardiocytes at 48h0.77 μg/ml (1.4 uM)
In-vitro rat hepatocytes at 24h>18.3 μg/ml (>33 uM)

EC50 for HepG2 cells at 72 h
GSH depletion at 24h25 uM
ATP depeletion>25 uM
CytoLite25 uM
Alamar Blue0.03 uM
LDH release0.01 uM

MEC for HepG2 cells at 72 h
ATP depeletion0.03 uM
CytoLite0.3 uM
Alamar Blue0.03 uM
LDH release0.03 uM

References:

-http://www.drugbank.ca/drugs/DB00997
-Tomoaki Inoue et al. "Predictive in vitro cardiotoxicity and hepatotoxicity screening system using neonatal rat heart cells and rat hepatocytes" AATEX 14, Special Issue, 457-462 Proc. 6th World Congress on Alternatives & Animal Use in the Life Sciences August 21-25, 2007, Tokyo, Japan
-Willem G. E. J. Schoonen, Walter M. A. Westerink and G. Jean Horbach, “High-throughput screening for analysis of in vitro toxicity”, Molecular, Clinical and Environmental Toxicology, 1:401-450, 2009.
-Willem G.E.J. Schoonen, Jeroen A.D.M. de Roos, Walter M.A. Westerink, Eric De´biton, “Cytotoxic effects of 110 reference compounds on HepG2 cells and for 60 compounds on HeLa, ECC-1 and CHO cells. II Mechanistic assays on NAD(P)H, ATP and DNA contents”, Toxicology in Vitro 19 (2005) 491–503.

Physical Properties ? Compound Assessment
Accepted and listed within the ToxCast/Tox21 program. ? No - Not Included in ToxCast Phase I and II Chemicals List.
Substance stability. ? Light sensitive [Doxorubicin hydrochloride Sigma Aldrich 44583 MSDS]

Stable for at least 1 year after receipt when stored, as supplied, at room temperature. Stock solutions are stable for up to 3 months at -20°C. Enzo Lifescience BML-GR319-0005 Product Datasheet

Doxorubicin hydrochloride is unstable in solutions with a pH less than 3 or greater than 7. Acids split the glycosidic bond in doxorubicin, yielding a red-colored, water insoluble aglycone (doxorubicinone, also known as adriamycinone) and a water soluble, reducing amino sugar (daunosamine). NLM Info

Soluble in buffer solution at 30 times the in vitro IC50 for toxicity. ? Doxorubicin hydrochloride soluble to 50mM in water Tocris Bioscience (2252) Product details

Doxorubicin hydrochloride Solubility in water> 50 mg/ml Sigma Aldrich 44583 Product details


estimated intrinsic solubility : 1.6283e-2 mg/ml
estimated solubility in pure water at pH 7.68: 0.271 mg/ml
estimated solubility in water at pH 7.4: 0.29 mg/ml
(Calculations performed using ACD/PhysChem v 12.0)

Solubility in DMSO 100 times buffer solubility. ? Doxorubicin hydrochloride soluble Sigma Aldrich 44583 Product details
Vessel binding properties. ? The stability of doxorubicin was determined in plastic and glass

administration containers over 48 hours. It was more stable in plastic (polyvinyl chloride) than in glass containers.

References:

-J Benvenuto, RW Anderson, K Kerkof, RG Smith, TL Loo. Stability and compatibility of antitumor agents in glass and plastic containers. Am J Hosp Pharm 1981, 88:1914-8
Vapor pressure. (Non-volatile) ? Estimated vapor pressure (25°C): 2.53E-23 mmHg (Calculation performed using EPI Suite v4.1)

Authors of this ToxBank wiki page

David Bower, Egon Willighagen
Retrieved from "http://wiki.toxbank.net/wiki/Doxorubicin"
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