Iba Final Lecture 1 Notes from the 23rd, continue polymorphism lecture When you see arrows in the

he notes pay attention to the sizes the contributions of particular pathways In the case of codeine O-demethylation of codeine which is catalyzed by p450 2D6 not inducible, genetically polymorphic contributes only 2-5% of hepatic p450 but contributes to about 20% metabolism of all drugs Very thick arrow -_> formation of morphine contributes only 10% of the total metabolism of codeine not enough but that very small amount of morphine formed is sufficient to account for the analgesic effect of codeine bioactivation pathway You want to notice that the major metabolic pathway for codeine metabolism is at other sites other than O-demethylation so the critical pathway as far as bioactivation of the drug to the pharmacologically active drug contributes only 10% Once you form morphine glucuronidation at the 6th position is an activation reaction glucuronidation can also occur at 3 position catalyzed primarily by the same glucuronidation that forms the one at 6th position pay attention to arrows Glucuronidation at the 6th position is an activation, not a detoxification demethylation is an activation reaction

most of these reactions occur in the liver very important how morphine glucuronide gets from the liver into the brain one of the factors you need to consider is that CYP 2D6 and UGT 2B7 are highly localized in certain neurons in the brain also need to remember that there are some transporters in the CSF -BBB interface that acts as OATs some people thing that all of these activation reactions occur within the brain, and a lot of ppl think that glucuronide is actively taken up from the periphery into the brain so these are questions for you to think about

the next polymorphism GST Major detoxification enzyme -> plays critical role in the detoxification of many chemicals Certain bioactivation reactions catalyzed by GST remember what class of chemicals are activated by GST Small molecular weight alkyl, alkylated solvents, methylene chloride, When you conjugate those compounds with glutathione they lead to bioactivation Halogens when you have carbon linked to these groups that glutathione will conjugate compounds that have NO2 function, halogens, bromides, iodides, and chlorides Polymorphism in GST GST has 8 different classes alpha, mu, theta, pi, zeta,sigma,kappa,and omega 5 of these classes are known to be genetically polymorphic Deficiencies/Mutations in these classes causes total absence of the enzyme absence of GST

GST pi, and GST mu One consequence of GST pi deficiency individuals deficient in this enzyme and have received chemotherapy therapy-related myelogenous leukemia t-AML massive deletion(?) tahts caused by the chemotherapeutic agents cyclophosphamide (activated by CYP 2B6 to phosphoramide mustard (chemotherapeutic agent) and then acrolein (very toxic) Cyclophosphamhide is a prodrug, widely used chemo agent, forms toxic intermediates almost all chemotherapeutic agents target most tumor cells and normal cells most individuals all the reactive intermediates are detoxified by GST normal level GST, still have sufficient amount of reactive intermediates to target tumors if you lack GST, the system is all screwed up the toxic intermediates starts damaging regular cells toxicity is much more pronounced in individuals with deficiency in GST compared to subjects with normal levels of GST Remember cyclophosphamide is a prodrug and requires bioactivation for the therapeutic drug Phospharamide targets tumor cells and then the excess of both acrolein and phosphoramide are detoxified by GST when you lack this enzyme , toxicity Need some level of GST to get rid of excess toxic intermediates

Susceptibility to Aflatoxin-induced liver cancer Once you form the epoxide the 2 fates to the epoxide 1. Binds to DNA to initiate mutations that may lead to all kinds of toxic effects including cancer 2. detoxification by GSTM1, so individuals deficient aflatoxin-induced liver cancer is higher same thing applies to Cigarettes smoking Incidences of lung cancer from smoking is higher in subjects who are deficient in GSTM1 benz__iamine..(?_..--> major carcinogen in tobacco smoke requires bioactivation by epoxide, once you form the epoxide 1. react with DNA 2. detoxified by GSTM1 smokers deficient in GSTM1 higher probability of developing lung cancer compared to smokers with normal levels of enzyme just like all of the other polymorphisms different in ethnic distributions , were just comparing African Americans and Caucasians when you see GST *0 mutation that results in total absence of that enzyme, none at all Polymorphism in transporters Only 2 of them MDR1 (p glycoprotein), and MRP2 transporters (involved in export of conjugated bilrubinin the liver) MDR1 mutations clinical consequences, and some arent one notable mutation mutation in exon 26 in the gene encoding MDR1 thymidine substituting for cytosine nucleotide sequence C3435

Loss of function mutation individuals who are homozygous for the mutation will have impaired ability to efflux drugs in the GIT cells and also have lowered ability to export drugs from the liver into the bile consequence is the increased bioavailability of drugs in these individuals In addition to the liver MDR1 are also present in lymphocytes important clinical significance in antiretroviral therapy in treatment of HIV/AIDS these patients who have the loss of function mutation are more responsive to antiretroviral therapy than the wildtype MDR1 because the drug is more bioavailable So as far as the general effect what you need to remember is that the transporters are very redundant really no clear evidence that a mutation in MDR1 is associated with serious clinical deficiency mutation in one of the transporters, can be taken up by other transporter of some sort The only drug thats known to exhibit differences in plasma levels due to mutation in MDR1 is this drug Digoxin used to be used for treating CHF very dangerous drug, very very toxic because polymorphisms of these transporters MDR1 polymorphism is a predictor of antiretroviral response HIV/AIDS drugs

Next polymorphism MRP2 MRP2 Responsible for efflux of conjugated bilirubin from the liver into the bile deficiency in this transporter MRP2 is responsible for pumping out organic anions Deficiency Dubin-Johnson syndrome Deficiency in transporters of conjugated bilirubin, whereas the other 2 syndromes discussed last exam are deficiencies in the conjugation reactions Former conjugated hyperbiliruinemia (DJ) and the latter 2 syndromes unconjugated hyperbilrubinemia know the difference between the 2 Review (pasted from slides) Two types of the genetic abnormality are known: Gilberts syndrome: a common, mild (and often benign) disorder (characterized by 60-70% increase in plasma unconjugated); has a US population incidence of 5-10%; results from mutations in the UGT1A1 promoter, leading to lower than normal level of hepatic UGT1A1 protein. a chronic, common, mild (and often benign) defect in hepatic bilirubin conjugation and clearance (jaundice); circulating serum bilirubin is 60-70% higher than normal has a 5-10 % incidence in the US population (not a rare disorder) results from dinucleotide (TA) repeat (insertion) polymorphism in the UGT1A1 promoter, leading to qualitatively normal but quantitatively lower than normal level of UGT1A1 (a regulatory sequence polymorphism).

 Crigler-Najjar (CN) syndrome a rare, complete deficiency in (type I CN) or severely diminished level of (type II CN) hepatic UGT1A1 activity. diminished to absent UGT1A1 protein results from coding sequence polymorphisms in the UGT1A1 gene. a rare, complete (and potentially lethal) deficiency in hepatic UGT1A1 activity (type I) or a drastically diminished level of the enzyme (type II) -in Crigler-Najjar type II, low levels of bilirubin glucuronides are detected in duodenal secretions diminished to absent levels of UGT1A1 protein results from mutations in the open reading frames of the UGT1A1 gene (coding sequence polymorphism) DJ

Associated with at least 15 polymorphisms 10 different variants different contributions of the alleles so the severity of the deficiency ranges from very mild to very severe Conjugated hyperbilirubinemia formed conjugates but are not exported.