Iba Final Lecture 3 Discussing Lipid peroxidation from previous Benzene (Snyders) Lecture

Contain an alpha methylated carbon carbon that contains 2 hydrogens and they are separated by 2 double bonds these hydrogens are the most reactive, they are the most susceptible to abstraction by hydroxyl radical that comes from the Haber Weiss reaction will abstract this hydrogen from the polyunsaturated fatty acid to form a glycol very reactive and it will react with molecular oxygen to form a fatty acid peroxide --> toxic radical As you can see any of these glycols -- > once is formed, will come around and abstract a hydrogen from another intact lipid so this reaction will continue until you exhaust all of the fatty acids that are present in membranes By immobilizing the ion with EDTA you can prevent the formation of the radical Once the radicals are formed if you have the compound known as Butylated Hydroxyl Toluene BHT antioxidant inactivates the fatty acid radicals, the peroxyl radical or the hydroxyl radical

Lipid Peroxidation - Exhaustive oxidation of fats by mechanisms involving free radicals Continue with Enzyme induction Lecture As said last lecture, Almost all the genes that contain the xenobiotic response element thats recognized between any receptor and the ligand inducer will be activated. This is a very brief example of all the genes that will respond to the Xenobiotic response element So these are the clinical consequences of drug metabolism and induction by tobacco smoke happens to be of these genes that contain the xenobiotic response element Benzpyrene which is a polycyclic aromatic hydrocarbon happens to be one of the important major inducer ligands that activates the receptor and produces all the responsive genes one of which is CYP 1A2 in the liver Phenacetin used previously as a prodrug for Tylenol APAP because o-denethylation by 1A2 formed APAP Previously was analgesic until it was discovered that it caused severe kidney problems In a hospital, for 7 days then they give the patient a dose of orally administered Phenacetin, and monitored the kinetics over time

The same population put on diet of steak put on an open flame and broiled and gave them the same dose of phenacetin the plasma levels of phenacetin or the area under the curve was significantly lower in the same patient that was eating charbroiled steak compared to when they were eating a regular diet there was an induction of the metabolism of phenacetin the induction was due to benzpyrene when you have a combustion of any organic materials leads to the formation of polycyclic aromatic hydrocarbons and the benzpyrene is all broiled steaks, hot dogs, anything you cook on an open flame combustion leads to formation of Cyp 1A inducing products so these are some of the findings, as you can see the effects of cigarette smoke smoking tobacco (organic material) the burning, the combustion is associated with polycyclic aromatic carbons speaks about inducers Pasted from slide Representative inducers include dioxin (an environmental pollutant found in defoliants, paper mills, etc), benzo(a)pyrene (found along with other polycyclic hydrocarbons [in smog and combustion emissions (including tobacco smoke)], indole-3-carbinol (found predominantly in cruciferous vegetablesbroccoli, brussel sprout, cabbage, etc.), omeprazole (an anti-ulcer drug). chewing tobacco does not have this effect phenacetin the plasma level is significantly decreased in smokers compared to controls theophylline drug used for asthma metabolism of theophylline is induced as shown by the decrease in plasma levels of smokers in compared to smokers warfarin metabolism is also induced note that this warfarin metabolism refers to the overall plasma concentration of warfarin S is metabolized specifically be 2C9 whereas R is metabolized by 1A2 as well as others all these drugs appreciate the fact the exposure to these inducing agents will lower the plasma levels of these drugs dont need to memorize the drugs in the table just need to know the trends, this table you do need to know This is the summary of the different inducers of p450s at least to 72 familes of p450s only those in the families 1-4 are involved in drug metabolism --> the rest do not play a major role in drug metabolism not going to talk about them much at all although mention mechanisms the nuclear receptors the AhR aryl hydrocarbon receptor is not a true nuclear receptor it can be activated as a transcription factor but

its not classified as a nuclear receptor dimerizaton with partner ARNT these 2 plus an inducer are required for the translocation into the nucleus for the next class of enzymes CYP 2B induction by this group only group that is also associated with induction of CYP reductase so for these inducers the barbiturates, rifampin (antimicrobial), hyperforin (St. Johns Wort) these inducers interact with the nuclear receptor that sknown as Constiuitive Androstane Receptors (CAR) so the dimerization partner is RXR so the binding of the inducers then the 3A class along with thte transporters (pgps) the regulation of pgps 3A4, 2C9 and several other transporters of the are all induced by the same barbiturates, steroids, rifampin, hyperforin that receptor that is involved in regulating these snzymes is known as the Pregnane X receptor results from the fact that preg__one which is an endogenous steroid is one of the most potent inducers of this class of enzymes the dimerization of the enzyme is the same as the induction of the 2B enzyme RXR 4A mostly involved in the metabolism of fatty acids nuclear receptor is PPAR and it has the same dimerization as the other 2 the potent inducers of 4A are the fibrates dont need to pay much attention to this class so each class 1-4 the differences are essentially in the nuclear receptors some of them share the same dimerization partner whereas ARNT is specific for the 1A receptor should be familiar with these differences so for the induction of this class, 2B members CAR the induction of this particular enzyme is different from others the CAR is a nuclear receptor, exists in an inactive form bound to an endogenous compound which is an inverse agonist lowers the baseline levels has functions opposite to that of an agonist you can call it an endogenous inhibitor nuclear receptor exists inactivated by an inverse agonist this is why its called CAR this is a mechanism of repression of genes gene is actively repressed, this is one of the examples inverse agonist would cause opposite, attain effect on baseline level agonists and inverse agonists both have intrinsic activity whereas antagonists do not

they interact with nuclear receptors and elicit a response the ligand enters the nucleus it competes and displaces the inverse agonist gene is activated leads to transcription of all these genes including CYP reductase which is seldom induced by other inducers the gene is constitutively repressed and then when an agonist displaces the endogenous repressor activates gene transcription

2B6 mostly refer to detailed slide HIV/AIDS Cyclophosphamide chemotherapeutic drug that requires metabolic activation 2B6 required loss of function mutation of 2B6 will not benefit from the therapeutic effects of cyclophosphamide cyclophosphamide induces 2B6 this is the phenomenon known as autoinduction refers to the ability of the drug to induce its own metabolism monitor the patient carefully to up adjust the dose to maintain therapeutic benefits the other side of the point is that not all drugs that induce enzymes are metabolized by enzymes induced by those drugs tries to reword if the drug metabolizes an enzyme doesnt necessarily mean that that enzyme will enhance the metabolism of the drug not all drug inducers are substrates for the enzymes that they induce drug A might induce enzyme B but the enzyme B may or may not metabolize A to product A1 just because a drug induces an enzyme -_. Does not mean that that enzyme will metabolize the drug that induced it. So when the drug induces its own metabolism auto-induction does not always happen, An enzyme does not always induce its own metabolism, may induce DMEs, but that enzyme might not metabolize that particular drug Drugs that are substrates there are drugs that are inhibitors do not need to be metabolized by the enzymes noncompetitive inhibitors Go through different clinical situations and see what will happen when you administer 2 drugs and one of them extensively metabolize because of the resulting induction have an appreiciation for the plasma levels and the need to adjust the dose

3A and transporters

now for the induction for the 3A class and the different transporters Pregnane/Steroid 3A is involved in the metabolism of endogenous steroids particularly preg__lone is a potent inducer PXR gets into the cell and interacts with the nuclear receptors extranuclear ligand-activated transcription factors only gets into the nucleus after theyre activated by the ligands ligand-receptor complex gets translocated into the nucleus dimerization DR3 responsive element is different for all receptor complexes, dont need to know leads to transcriptional activation of all of these responsive genes 3A4, 3A5, pgps, and other PXR responsive genes Progesterone is a major endogenous inducer of 3A

Post-Transcriptional Induction of Drug-Metabolizing Enzymes Most pronounced for 2E1 note that you cannot have the enzyme in the first place if there was no post-transcriptional induction Almost all the chemicals that increase the cellular levels of 2E1 do so by affecting induction in the post-transcriptional level One mechanism is stabilizing the transcript, increasing the half-life of mRNA or increasing the translation of the mRNA and one way to increase the half-life of mRNA is by inhibiting the degradation by mRNAse accumulation of mRNA, more translation of more protein Another mechanism is by stabilizing and decreasing the rate of degradation of the protein by inhibiting protease , ubiquitination all enzymes have a finite half life (?) and are usually degraded by specific proteases So the inducers of 2E1 inhibit the proteases and the mRNAases 2E1 Inducers include organic solvents: benzene, toluene, acetone uncontrolled diabetes is known to increase hepatic levels of 2E1 due to increased levels of ketone bodies The next chart is just a summary and as you can see theres quite an overlap among the different CYP inducers the important thing is to note As much as you should know what nuclear receptors are involved with these specific p450s , theres no really no reason to memorize the different classes of drugs know ethanol, antiepileptic drugs, just bolded/emphasized Some notable points refer mostly to detailed slide Tobacco very potent inducer If the inducer is a substrate for the enzyme it induces and you have another competitive substrate the inducer will inhibit the activity of the enzyme acute

Its different if the patient has been exposed to the inducer for prolonged period of time and then exposed to the drug because the induction refers to the accumulation of the enzyme --chonic So chronic effect increased metabolism of the drug Acute decreased metabolism these are the different scenarios which you should be familiar with

CYP 3A4 kinetics Refer mostly to slide Felodipine is an antiarrhythmic it is a very toxic compound with a very NTI Below 2 nanomolar concentration the drug is ineffective, and above 4 causes severe arrhythmia Refer to slide