Cytochrome P450 1A1-mediated anticancer drug discovery: in silico findings

Introduction: Target-specific drugs may offer fewer side/adverse effects in comparison with other anticancer agents and thus save normal healthy cells to a greater extent. The selective overexpression of cytochrome P450 1A1 (CYP1A1) in tumor cells induces the metabolism of benzothiazole and aminoflavone compounds to their reactive species, which are responsible for DNA adduct formation and cell death. This review encompasses the novelty of CYP1A1 as an anticancer drug target and explores the possible in silico strategies that would be applicable in the discovery and development of future antitumor compounds.

Areas covered: This review highlights the various ligand-based and target-based in silico methodologies that were efficiently used in exploration of CYP1A1 as a novel antitumor target. These methodologies include electronic structure analysis, CoMFA studies, homology modeling, molecular docking, molecular dynamics analysis, pharmacophore mapping and quantitative structure activity relationship (QSAR) studies. It also focuses on the various approaches used in the development of the lysyl amide prodrug of 5F-203 (NSC710305) and dimethanesulfonate salt of 5-aminoflavone (NSC710464) as clinical candidates from their less potent analogues.

Expert opinion: Selective overexpression of CYP1A1 in cancer cells offers tumor-specific drug design to ameliorate the current adverse effects associated with existing antitumor agents. Medicinal chemistry and in vitro driven approaches, in combination with knowledge-based drug design and by using the currently available tools of in silico methodologies, would certainly make it possible to design and develop novel anticancer compounds targeting CYP1A1.     

Cytochrome P450 1A1-mediated anticancer drug discovery: in silico findings

Introduction: Target-specific drugs may offer fewer side/adverse effects in comparison with other anticancer agents and thus save normal healthy cells to a greater extent. The selective overexpression of cytochrome P450 1A1 (CYP1A1) in tumor cells induces the metabolism of benzothiazole and aminoflavone compounds to their reactive species, which are responsible for DNA adduct formation and cell death. This review encompasses the novelty of CYP1A1 as an anticancer drug target and explores the possible in silico strategies that would be applicable in the discovery and development of future antitumor compounds. Areas covered: This review highlights the various ligand-based and target-based in silico methodologies that were efficiently used in exploration of CYP1A1 as a novel antitumor target. These methodologies include electronic structure analysis, CoMFA studies, homology modeling, molecular docking, molecular dynamics analysis, pharmacophore mapping and quantitative structure activity relationship (QSAR) studies. It also focuses on the various approaches used in the development of the lysyl amide prodrug of 5F-203 (NSC710305) and dimethanesulfonate salt of 5-aminoflavone (NSC710464) as clinical candidates from their less potent analogues. Expert opinion: Selective overexpression of CYP1A1 in cancer cells offers tumor-specific drug design to ameliorate the current adverse effects associated with existing antitumor agents. Medicinal chemistry and in vitro driven approaches, in combination with knowledge-based drug design and by using the currently available tools of in silico methodologies, would certainly make it possible to design and develop novel anticancer compounds targeting CYP1A1.     

Cytochrome P450 interactions in human cancers: new aspects considering CYP1B1

Molecular epidemiological studies are now a powerful tool to determine differential genetic susceptibilities to cancer-causing agents, and to obtain information on potential mechanisms. Cytochrome P450 (CYP) allelic variants are considered biomarkers of susceptibility to cancer. Such variants have an influence on the bioactivation and thereby on the potency of chemical carcinogens. This is very much straight forward for tobacco smoke-related human cancers. A new aspect is the implication of CYP1B1 in tobacco smoke-related cancers at several organ sites. On this basis, the present review is focused on lung, breast, urinary bladder and head and neck cancer. The CYP profile of the human lung includes CYP1A1, -1B1, -2A6, -2A13, -2B6, -2C18, -2E1, -2F1, -3A5 and -4B1. Polycyclic aromatic hydrocarbons (PAHs) and nitrosamines, as active components of tobacco smoke, appear as primary chemical factors for lung malignancies. For human mammary cancer, the use of hormone replacement therapy (HRT) has been shown to be associated with an increase of breast cancer risk, and there seems to be a link between risks caused by HRT use and modifying polymorphisms of drug/xenobiotic enzymes. Specifically, an association of the CYP1B1*3/*3 genotype with increased breast cancer risks has been postulated. Cigarette smoking is a major cause of human urinary bladder cancer. Arylamines, PAHs and nitrosamines are locally activated within the urothelium. Important CYPs in the bladder epithelium of experimental animals and man are CYP1B1 and -4B1. Alcohol consumption and tobacco smoking are known as the major causes of head and neck cancers. Recently, it appears that a polymorphic variant CYP1B1*3/*3 relates significantly to the individual susceptibility of smokers to head and neck cancer, supporting the view that PAH are metabolically activated through CYP1B1. It appears that CYP1B1 plays a key role for the activation of carcinogens at several organ targets, with a likelihood of complex gene-environment interactions implying Phase II enzymes.     

Cytochrome P450 and anticancer drugs

Cytochrome P450 (CYP) is involved in the metabolism of a variety of anticancer drugs. CYP activities are known to be modified by several factors including genetic polymorphisms, changes in physiological conditions such as age, disease status or intake of certain drugs or foods or environmental factors such as smoking. These factors may cause interindividual differences in the pharmacokinetic profiles of anticancer drugs, leading to the variations of efficacy or toxicity of the drugs. Genetic polymorphisms present in CYPs sometimes result in the reduced activity of the enzymes causing low metabolic clearance of drugs or low production of active metabolites. For example, the formation of endoxifen, which is an active metabolite of tamoxifen, was less in patients with inactive polymorphic CYP2D6 than those with the wild type enzyme. CYP3A is the most abundant CYP expressed in the human liver and the small intestine that is involved in the metabolism of various anticancer drugs. The catalytic activity of CYP3A shows a large interindividual variability giving rise to large interindividual differences in the pharmacokinetic profiles of some anticancer drugs. So far, many attempts have been made to monitor the phenotypic activity of CYP3A in order to reduce the pharmacokinetic variations of anticancer drugs. Erythromycin, midazolam and cortisol are commonly used to monitor in vivo hepatic CYP3A activity. These methods have been applied to reduce the pharmacokinetic variations of docetaxel. Drug-drug interactions related to CYPs also modulate the pharmacokinetic profiles of anticancer drugs. These factors should be considered when trying to optimize and individualize chemotherapy.     

Cytochrome P450 (CYP) in fish

Cytochrome P450 (CYP) enzymes are members of the hemoprotein superfamily, and are involved in the mono-oxygenation reactions of a wide range of endogenous and exogenous compounds in mammals and plants. Characterization of CYP genes in fish has been carried out intensively over the last 20 years. In Japanese pufferfish (Takifugu rubripes), 54 genes encoding P450s have been identified. Across all species of fish, 137 genes encoding P450s have been identified. These genes are classified into 18 CYP families: namely, CYP1, CYP2, CYP3, CYP4, CYP5, CYP7, CYP8, CYP11, CYP17, CYP19, CYP20, CYP21, CYP24, CYP26, CYP27, CYP39, CYP46 and CYP51.We pinpointed eight CYP families: namely, CYP1, CYP2, CYP3, CYP4, CYP11, CYP17, CYP19 and CYP26 in this review because these CYP families are studied in detail. Studies of fish P450s have provided insights into the regulation of P450 genes by environmental stresses including water pollution. In this review, we present an overview of the CYP families in fish.     

Cytochrome P450-mediated cardiovascular drug interactions

INTRODUCTION: There are numerous drug-drug interactions (DDIs) related to cardiovascular medications and many of these are mediated via the cytochrome P450 (CYP) system. Some of these may lead to serious adverse events and it is, therefore, essential that clinicians are aware of the important interactions that occur. AREAS COVERED: An extensive literature search was performed to analyze the CYP-mediated cardiovascular DDIs that lead to a loss of efficacy or potential toxicity. Cardiovascular drugs may be victims or act as perpetrators of DDIs. The paper analyzes CYP-mediated drug interactions concerning anticoagulants, antiplatelet agents, antiarrhythmics, β-blockers, calcium antagonists, antihypertensive medications, lipid-lowering drugs and oral antidiabetic agents. EXPERT OPINION: Cardiovascular DDIs involving the CYP system are numerous. Additionally, the spectrum of drugs prescribed is constantly changing, particularly with cardiovascular diseases and it is not necessarily the case that drugs that had shown safety earlier will always show safety. Clinicians are encouraged to develop their knowledge of CYP-mediated DDIs so that they can choose safe drug combination regimens, adjust drug dosages appropriately and conduct therapeutic drug monitoring for drugs with narrow therapeutic indices.     

Genetic variation of Cytochrome P450 1B1 (CYP1B1) and risk of breast cancer among Polish women

Four single nucleotide polymorphisms (SNPs) in CYP1B1 (Ex2 + 143 C > G, Ex2 + 356 G > T, Ex3 + 251 G > C, Ex3 + 315 A > G) cause amino acid changes (R48G, A119S, L432V and N453S, respectively) and are associated with increased formation of catechol estrogens; however, epidemiologic evidence only weakly supports an association between these variants and breast cancer risk. Because genetic variability conferring increased susceptibility could exist beyond these putative functional variants, we comprehensively examined the common genetic variability within CYP1B1. A total of eight haplotype-tagging (ht)SNPs (including Ex3 + 315 A > G), in addition to two putatively functional SNPs (Ex2 + 143 C > G and Ex3 + 251 G > C), were selected and genotyped in a large case-control study of Polish women (1995 cases and 2296 controls). Haplotypes were estimated using the expectation-maximization algorithm, and overall differences in the haplotype distribution between cases and controls were assessed using a global score test. We also evaluated levels of tumor CYP1B1 protein expression in a subset of 841 cases by immunohistochemistry, and their association with genetic variants. In the Polish population, we observed two linkage disequilibrium (LD)-defined blocks. Neither haplotypes (global P-value of 0.99 and 0.67 for each block of LD, respectively), nor individual SNPs (including three putatively functional SNPs) were associated with breast cancer risk. CYP1B1 was expressed in most tumor tissues (98%), and the level of expression was not related to the studied genetic variants. We found little evidence for modification of the estimated effect of haplotypes or individual SNPs by age, family history of breast cancer, or tumor hormone receptor status. The present study provides strong evidence against the existence of a substantial overall association between common genetic variation in CYP1B1 and breast cancer risk.     

Cytochrome P450-activated prodrugs: targeted drug delivery

Cytochrome P450 (CYP) enzymes are a superfamily of heme containing proteins that catalyze xenobiotic metabolism phase I reactions. Oxidation reactions are the most common CYP-catalyzed reactions for both endogenous substrates and exogenous compounds, including drugs, although CYP enzymes are capable also to catalyze reduction reactions. Whereas the majority of clinically used drugs are inactivated by CYPs, several prodrugs are bioconverted to their active species by these enzymes. Therefore, this mechanism could be exploited to a greater extend, e.g. by taking advantage of the different CYP enzymes to achieve targeted drug delivery, to improve efficacy or to decrease the unwanted adverse effects of existing and novel drug molecules. This review describes the potential of CYP enzymes in prodrug design and summarizes a wide variety of CYP-activated prodrug structures, which are on the market or under the development. The bioactivation mechanisms of each CYP-activated prodrug structure are described and the specificity for the different forms of CYP enzymes is discussed.     

细胞色素P450 1B1在上皮性卵巢癌组织中的表达

目的：检测卵巢癌组织、卵巢良性上皮性肿瘤及正常卵巢组织中细胞色素P4501B1（cytochrome P4501B1，CYP1B1）的表达，为以CYP1B1为靶点进行卵巢癌治疗提供理论基础。方法：采用免疫组化SP法检测53例上皮性卵巢癌、30例卵巢良性上皮性肿瘤及19例正常卵巢组织中CYP1B1的表达。结果：53例卵巢癌中CYP1B1阳性率92．45％，明显高于卵巢良性肿瘤组织（13．33％），差异有统计学意义（P〈0．01）。正常卵巢组织CYP1B1呈阴性表达。14例同时收集的卵巢癌转移灶组织有13例CYP1B1阳性表达，阳性率为92．86％。结论：CYP1B1可作为一个抗癌药开发及逆转化疗药耐药的新靶点。     

Cytochrome P450 (CYP) expression in human myeloblastic and lymphoid cell lines

To explore the physiological roles of cytochrome P450 (CYP) in peripheral blood cells, we examined which isoforms of CYP families were expressed in human myeloid leukemia cell lines (U937, HL-60 and K562) and lymphoid cell lines (BALL-1, MOLT-4 and Jurkat) by RT-PCR. We observed relatively high expression of CYP1A1, CYP1B1, CYP2A6, CYP2A7, CYP2D6, and CYP2E1 in all cell types, but CYP2A13 and CYP2C9 expression was not detected. Expressions of aryl hydrocarbon (Ah) receptor and Ah receptor nuclear translocator (ARNT), which mediate induction of the CYP1 family, were also detected in all cell types. Cell-type specific expression of CYP3A4 and CYP3A5 was observed in MOLT-4 and K562 cells. Weak, but significant, expression of CYP3A7 was detected in K562 cells. The profile of CYP expression in the culture cells reported here provides information that furthers our understanding of the physiological roles of CYP enzymes in human blood cells.     

Cytochrome P450 3A: ontogeny and drug disposition

The maturation of organ systems during fetal life and childhood exerts a profound effect on drug disposition. The maturation of drug-metabolising enzymes is probably the predominant factor accounting for age-associated changes in non-renal drug clearance. The group of drug-metabolising enzymes most studied are the cytochrome P450 (CYP) superfamily. The CYP3A subfamily is the most abundant group of CYP enzymes in the liver and consists of at least 3 isoforms: CYP3A4, 3A5 and 3A7. Many drugs are mainly metabolised by the CYP3A subfamily. Therefore, maturational changes in CYP3A ontogeny may impact on the clinical pharmacokinetics of these drugs. CYP3A4 is the most abundantly expressed CYP and accounts for approximately 30 to 40% of the total CYPcontent in human adult liver and small intestine. CYP3A5 is 83% homologous to CYP3A4, is expressed at a much lower level than CYP3A4 in the liver, but is the main CYP3A isoform in the kidney. CYP3A7 is the major CYP isoform detected in human embryonic, fetal and newborn liver, but is also detected in adult liver, although at a much lower level than CYP3A4. Substrate specificity for the individual isoforms has not been fully elucidated. Because of large interindividual differences in CYP3A4 and 3A5 expression and activity, genetic polymorphisms have been suggested. However, although some gene mutations have been identified, the impact of these mutations on the pharmacokinetics of CYP3A substrates has to be established. Ontogeny of CYP3A activity has been studied in vitro and in vivo. CYP3A7 activity is high during embryonic and fetal life and decreases rapidly during the first week of life. Conversely, CYP3A4 is very low before birth but increases rapidly thereafter, reaching 50% of adult levels between 6 and 12 months of age. During infancy, CYP3A4 activity appears to be slightly higher than that of adults. Large interindividual variations in CYP3A5 expression and activity were observed during all stages of development, but no apparent developmental pattern of CYP3A5 activity has been identified to date. Profound changes occur in the activity of CYP3A isoforms during all stages of development. These changes have, in many instances, proven to be of clinical significance when treatment involves drugs that are substrates, inhibitors or inducers of CYP3A. Investigators and clinicians should consider the impact of ontogeny on CYP3A in both pharmacokinetic study design and data interpretation, as well as when prescribing drugs to children.     

肿瘤组织中的CYP1B1及其抑制剂的抗肿瘤活性

综述细胞色素P450酶（CYP）1B1在肿瘤组织中的表达、在肿瘤的发生发展和诊断与干预中的作用以及其抑制剂的研发和抗肿瘤活性。CYP1B1在正常组织中低表达,而在许多肿瘤组织中则特异性高表达,可激活和代谢产生致癌物质,并可致多种抗癌药物代谢失活而使肿瘤耐药,因此它既可用于早期癌症的诊断,又可作为理想的抗肿瘤作用靶点而用于药物研发。     

Bioflavonoids: selective substrates and inhibitors for cytochrome P450 CYP1A and CYP1B1

Interactions of six naturally occurring flavonoids (acacetin, diosmetin, eriodictyol, hesperetin, homoeriodictyol, and naringenin) with human cytochrome P450 (CYP1) enzymes were studied. The flavones acacetin and diosmetin were potent inhibitors of ethoxyresorufin O-dealkylase (EROD) activity of CYP1A and CYP1B1. Hydroxy and/or methoxy substitutions at the 3' and 4' positions in the flavonoid structures were the major factors involved in conveying selectivity for the different cytochrome P450 enzymes. Eriodictyol, homoeriodictyol and naringenin were very poor inhibitors of human CYP1A EROD activity (IC(50)4 microM). Hesperetin and homoeriodictyol selectively inhibited human CYP1A1 and CYP1B1. Only homoeriodictyol selectively inhibited human CYP1B1 (IC(50) 0.24 microM). Hesperetin was O-demethylated by both human CYP1A1 and 1B1 to eriodictyol, which was then further metabolized by the same enzymes. Hesperetin was not metabolized by human CYP1A2 or CYP3A4.     

Cytochrome P450 CYP1B1 and catechol O-methyltransferase (COMT) genetic polymorphisms and breast cancer susceptibility in a Turkish population

Epidemiological studies indicate that most risk factors for breast cancer are related to reproductive and hormonal factors. Estrogen has been proposed to trigger breast cancer development via an initiating mechanism involving its metabolite, catechol estrogen (CE). Because of the important role of cytochrome P450 1B1 ( CYP1B1) and catechol O-methyltransferase ( COMT) in mammary estrogen and carcinogen metabolism, we examined the CYP1B1 and COMT genes to determine whether genetic variations could account for inter-individual differences in breast cancer. In this case-control study, we determined CYP1B1 and COMT genotypes in 84 breast cancer patients and 103 healthy unrelated women controls from a Turkish population. In the case of CYP1B1, we genotyped CYP1B1*3 (L432 V) allele. We found that carriers of the CYP1B1*3 allele were more frequent among breast cancer patients with adjusted odds ratio (OR) for age, age at menarche, age at first full-term pregnancy, body mass index (BMI) and smoking status of 2.32 (95% confidence interval 1.26-4.25) associated with the allele. However, this allele appeared to be a significant factor for susceptibility only in patients with a BMI greater than 24 kg/m(2). Menopausal status did not appear to affect susceptibility. In the case of COMT, there was no significant difference in susceptibility for breast cancer development between patients with low activity COMT-L (V158 M) allele and high activity COMT-H allele, and susceptibility was not affected by menopausal status, BMI or CYP1B1 genotype. We conclude that the CYP1B1* 3 allele appears to be a factor for susceptibility to breast cancer in Turkish women especially those with a BMI greater than 24 kg/m(2).     

振源胶囊对细胞色素P_(450)酶CYP1A2、CYP3A4、CYP2E1的影响

目的:研究振源胶囊对细胞色素P450酶CYP1A2、CYP3A4、CYP2E1的影响。方法:用Cocktail探针药物法,将Wistar大鼠随机分组,灌胃给予振源胶囊溶液,以生理盐水组为空白对照,诱导10d,于股动脉插管,注射给予3种探针药物咖啡因、氨苯砜、氯唑沙宗,通过高效液相色谱法检测各探针药物的代谢率来评价各组CYP1A2、CYP3A4、CYP2E1亚型酶的活性;药动学计算采用DAS2.0软件完成。结果:给予振源胶囊的大鼠,咖啡因代谢加快,半衰期缩短;氨苯砜代谢减慢,半衰期延长;氯唑沙宗半衰期与空白对照组比较无显著差异(P>0.05)。结论:振源胶囊对大鼠CYP1A2有诱导作用,对CYP3A4有抑制作用,对CYP2E1的作用不明显。     

Cytochrome P450 2B (CYP2B)-mediated activation of methyl-parathion in rat brain extracts.

The role of cytochrome P450 (CYP) and the CYP isoform involved in the activation of the widely used pesticide methyl-parathion (MePA) were investigated in rat brain extracts by measuring the effect of different CYP inhibitors on acetylcholinesterase (AChE) inhibition by MePA. Brain extracts provide a useful tool to study the activation mechanisms of organophosphorus compounds (OP) since they contain both the activating enzyme(s) and the molecular target for OP toxicity. As expected, in incubations of rat brain extract supplemented with NADPH, AChE activity was non-competitively inhibited by the presence of MePA, indicating that MePA was activated to its reactive metabolite methyl-paraoxon (MePO). Indeed, Vmax(app) decreased from 13.4 to 8.7 micromol thionitrobenzoic acid (TNB)/min per mg protein. MePA activation by rat brain extracts, as measured by the AChE inhibition produced by the presence of the pesticide in the incubation, was fully prevented by previously bubbling the incubation mix with CO, by the presence of monoclonal anti-rat CYP2B1/2B2 antibodies and by the addition of phenobarbital (PB), a CYP2B substrate. Interestingly, MePA showed a greater affinity for CYP2B than PB. CYP1A1 antibodies showed no effect on MePA activation. The presence of cytochrome P450 2B (CYP2B) in the rat brain extracts was confirmed by immunoblotting. These results demonstrate indisputably the responsibility of CYP2B in MePA activation in the rat brain in vitro, suggesting that metabolic activation of OP compounds in situ might be crucial for their organ specific toxicity to the central nervous system also in vivo.     

Cytochrome P450 (CYP) inhibition screening: comparison of three tests.

There are several different experimental systems for screening of in vitro inhibitory potency of drugs under development. In this study we compared three different types of cytochrome P450 (CYP) inhibition tests: the traditional single substrate assays, the fluorescent probe method with recombinant human CYPs, and a novel n-in-one technique. All major hepatic drug-metabolizing CYPs were included (1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, 3A4). Six compounds (sotalol, propranolol, citalopram, fluoxetine, oxazepam and diazepam) were selected for detailed comparisons. The IC50 values of each of these compounds were measured using the three assay types. The inhibitory potencies of these model drugs were generally within the same order of magnitude and followed similar inhibition profiles in all the assay types. Clinically observed inhibitory interactions, or lack thereof, were predictable with all three assays. Comparison of potencies of 'diagnostic' inhibitors revealed also some notable differences between the assays, especially regarding CYP2E1. The potency of inhibitors towards CYP3A4 was dependent on the substrate and reaction measured. Generally all three assays gave reasonably comparable results, although some unexplained differences were also noted.     

食物对细胞色素P450药物代谢酶的影响

细胞色素P450酶系在大多数内源性和外源性分子的生物氧化过程中发挥重要的作用,尤其在药物代谢方面。CYP450酶个体差异大,除了遗传因素的影响外,食物等外界因素也可能影响其活性或表达,从而影响经酶代谢药物的疗效和不良反应。故本文就食物因素对细胞色素P450酶影响的相关研究进行综述。