Assignment 1, HLTH 340, 2011 Winter

University of Waterloo
Environmental Toxicology and Public Health

Name: Samriti Mishra Student ID: 20315814 Professor Mccoll

HLTH 340: Environmental Toxicology and environmental health

Assignment 1

Question 1, Paraphrased: Given the chemical behavior of Nicotine (in Nicotine chewing gum for smoking cessation), the psychoactive constituent in cigarette smoke responsible for the euphoriant and addictive effects of chronic cigarette smoking, how does salivary pH variation (pH 6 to pH 8) affect the efficiency of it as an anti-craving agent? Explain your reasoning. How would you propose to ensure the optimal efficiency of nicotine absorption through the buccal membrane by controlling the salivary pH (the absorption method to have a useful pharmacological effect). The relative proportion of ionized and non-ionized nicotine for three pH levels: pH value pH 5.5 pH 7.4 pH 8.1 Ionized > 99 75.8 38.5 Non-Ionized <1 24.3 61.5

Response: When considering the toxicokinetics of Nicotine (a xenobiotic), absorption, distribution and metabolism would be affected by the relative proportion of ionized (hydrophilic) vs. non-ionized (lipophilic) nicotine. Like most drugs, absorption across the buccal membrane is the major source of absorption as it is without the risk of being degraded by gastrointestinal and hepatic first pass metabolism (Adrian, Olin, Dalhoff, Jacobsen, 2006). The rate limiting step in the buccal drug absorption process is the permeability across the buccal epithelium. Permeability of Nicotine depends on the partition coefficient of the non-ionized vs. ionized form. The non-ionized, lipophilic form would passively diffuse across the buccal membrane (having a partition coefficient Kow > 1), while the ionized, hydrophilic form would be not absorbed. Various in vitro studies confirm the increase in Nicotine absorption with increase in pH and the amount of non-ionized nicotine levels (Adrian et. al., 2006). Nicotine exists in equilibrium between the ionized and non-ionized form in aqueous media. The reaction shifts to the non-ionized form as the alkaline levels increased. The human salivary pH varied from pH 6 to pH 8, thereby resulting in varying degrees of absorption of the relative non-ionized concentration. The more alkaline the salivary pH, better would be the absorption of nicotine, resulting in reducing the cigarette cravings. Nicotine works by docking on to the cholinergic pathway and simultaneous release of acetylcholine, glutamate, dopamine and endorphins, resulting in over stimulation of the cholinergic pathways forming the rewards loop (Discovery health, 2001).To ensure that the salivary pH is kept to alkaline levels of pH greater than 8, sodium bicarbonate (alkaline salts) can be an additive to the nicotine gum. In this particular scenario, the NaOH(aq) would dissociate in the salivary aqueous media, giving up the hydroxide ion and increasing the alkaline nature of saliva. This would shift the nicotine recombination reaction to the right, making available the non-ionized, lipophillic form of nicotine, resulting in higher absorption rates.

HLTH 340: Environmental Toxicology and environmental health

Assignment 1

Ionized

Non-Ionized

Question 2, Paraphrased: Why would adding inorganic alkaline salts (eg. Ammonium sulfate) serve to alter the pharmacokinetic properties of the cigarette smoke and its psychoactive effects on the smoker. Why would the U.S. tobacco companies have chosen to introduce such additives in the first place? Solution: Adding inorganic alkaline salts to cigarettes would cause the addiction to dramatically go stronger. This is due to the fact that at alkaline pH, the lipophilic, non-ionized form of nicotine is present in the saliva at higher proportions. Alkaline media increases the lipophilic forms bio-availability thereby resulting in higher and faster absorption. Higher, speedy absorption correspondingly increases the blood plasma levels of nicotine, followed by uptake in the tissues. The cigarette smoke itself it acidic, so unless the alkaline additives are added, the bioavailability of nicotine (non-ionized, lipophilic form), the absorption of nicotine at the buccal membrane is quite limited. For nicotine to provide a strong psychoactive reaction, it must be delivered rapidly to the brain. The acute and repeated exposure stimulates the release of dopamine in the mesocorticolimbic dopamine system, an integral part of the brain reward system. Nucleus accumbens, a part of this system, is understood to play an important role in addiction. Rewards such as food and sex, along with emotions induced by music, is processes and regulated in this area. Nicotine exerts similar effects by activating specific sites called receptor proteins, which triggers release of dopamine in the nucleus accumbens and the secretion of other nerve-stimulating chemicals such as acetylcholine and glutamate in the hippocampus and cerebral cortex, which act to improve vigilance, attention and cognition. According to smokers, nicotine use helps them when they are depressed, stressed, embarrassed, bored, irritable or in a bad mood. Alleviating these unpleasant feelings with a cigarette helps reinforce the psychological aspects of tobacco addiction (The Cancer Council, 2011). In effect adding inorganic alkaline additives to cigarettes would act to promote addictive behavior, causing the consumer to seek out more cigarettes per day and eventually drive up the demands, resulting in financial profit. It should be noted that human beings have the capacity to develop tolerance to nicotine, thereby requiring increasingly more amounts of nicotine to attain the same high with the passage of time. This results in dangerous addictive behavior.

HLTH 340: Environmental Toxicology and environmental health

Assignment 1

Question 3, Paraphrased: The P-gp pump transporter is found in many cellular membranes, including the enterocytes of the small intestine, the liver hepatocytes and capillary endothelium cells comprising the blood-brain-barrier. Describe the role of the following xenobiotic chemicals and drugs on the activity of P-gp pump and the potential hazardous health effects this type of interaction might produce in humans: a) St Johns Wort (hyperforin) and cyclosporine: While St Johns wort is an anti-depression herbal medication (constituents are hyperforin and hypericin), cyclosporine is a drug that prevents organ rejections post-surgery. Cyclosporine is metabolized in the first pass elimination processes, both at the intestinal lining and hepatocytes). The P-gp pump pumps out the nonmetabolized small ratio of cyclosporine into the blood stream, from where it now distributes to various organs of the body. As mentioned earlier, hyperforin is an anti-depressant. Postsurgery, if a person intakes this to deal with the psychological stresses, strong drug interaction ensues. Hyperforin binds to the P-gp pumps, which make it unavailable for elimination of cyclosporine at the intestinal/hepatic levels (during first pass elimination) and at final uptake at the different organs (specially the blood brain barrier). This is results in high levels of cyclosporine in the blood plasma, which has then to be eliminated by kidney, which goes into hyper drive. This results in renal failure. b) Digoxin and erythromycin: Erythromycin is a macrolide antibiotic it suppresses or kills the bacteria by starving it of the important proteins necessary for its survival. Digoxin is a dirty dozen chemical, it is an omnipresent persistent organic pollutant and most human beings have been exposed to it. Ingested dioxin is reduced by the normal gut bacteria. Once this inactivated form is absorbed in the body, dioxin remains in the system sequestered in the adipose tissue. The half-life for dioxin is estimated to be 7 11 years. When erythromycin is ingested, it kills the normal bacterial flora of the digestive system as a side-effect. When dioxin is now consumed, its bioavailability is many folds the usual level in the blood stream. This exposure to dioxin can lead to a wide array of adverse health effects including cancer, birth defects, diabetes, learning and developmental delays, endometriosis, and immune system abnormalities (Agent Orange Association of Canada, 2008). c) Imodium (loperamide) and quinidine : Imodium is an oral anti-oral agent which slows down the motility of the intestinal wall, allowing the body to reabsorb any excess liquid (Johnson and Johnson Inc, 2010) . It acts on the medulla oblongata of the brain which controls the peristalsis of the intestines (along with a host of other involuntary functions like respiration and blood pressure). Quinidine on the other hand is an anti-arrhythmia medication. However, quinidine is also a P-gp inhibitor. When Imodium and quinidine are administered simultaneously, quinidine inhibits P-gp at the blood brain barrier, resulting in greater CNS penetration of loperamide and potentially causes serious neurotoxicity (Atkinson, 2007). As Imodium acts as a suppressant, and acts on the medulla oblongata of the hind stem. This results respiratory depression along with the desired effect of reduced rate of peristalsis.

HLTH 340: Environmental Toxicology and environmental health

Assignment 1

Question 4, Paraphrased: Rotenone is an insecticide, which has been implicated as a risk factor for Parkinsons disease in exposed individuals. Explain the chemical and biological properties in some exposed individuals, with special reference to MPTP. Discuss toxicokinetic and toxicodynamic factors that could contribute to neuronal damage to the motor systems of the brain. Why is this especially hazardous to children or younger persons? Solution: The following are the chemical structures of MPTP, MPP+ and Rotenone respectively

Neurotoxicity caused my MPTP was discovered as an serendipity when young cocaine addicts contracted Parkinsons disease. MPTP was present in the drug as a contaminant. MPTP is a lipophilic molecule (because of presence of benzene ring) which passes through the blood brain barrier and taken up by astrocytes, which are metabolically active cells. The astrocytes contain Mono-amine oxidase type B enzymes which convert sthe MPTP to the hydrophilic form MPP+. The now modified MPP+ is then taken up by the same dopaminergic reuptake protein channels as it MPP+ closely mimics dopamine. MPP+ then concentrates in the neuron, as there is no pump for MPP+ elimination. MPP+ enters the mitochondria and binds to complex 1 of the electron transport chain, subsequently causing lack of ATP formation. The dopaminergic neurons concentrate in the substantia nigra region of the brain, which is responsible for fine motor movements. A lack of energy in these neurons causes cells death, resulting in the progressive neurodegenerative disorder of Parkinsons. Consider the chemical structure of MPTP and Rotenone. Multiple benzene groups indicate lipophilicity of Rotenone, similar to MPTP. The Octanol/water partition coefficient of Rotenone as log PO/W is 4.10 (IPCS,2004). Further, the methoxy functional groups which are attached to the benzene ring are similar to the amine group in MPTP. This would cause the MAO-B enzyme to act similarly on Rotenone, causing to form the rotenone equivalent of MPP+. Reuptake mechanisms readily take up Rotenone (post enzymatic form) which is a molecular mimicry of dopamine. Rotenone proceeds to the mitochondria where is inhibits the transfer of electrons from iron-sulfur centers in complex I to ubiquinone. This interferes with NADH during the creation of usable cellar energy ATP. However, its not likely that a bioenergetics defect could account solely for rotenone-induced neuro-degeneration. Instead, rotenone toxicity may result from oxidative stress. Within complex I, upstream from the 4

HLTH 340: Environmental Toxicology and environmental health

Assignment 1

rotenone-binding site, is a site of electron leakage that produces ROA and impaired complex I activity enhances ROS formation. Rotenone exposure increases the protein carbonyls (indicators of oxidative stress by Reactive oxygen species) in midbrain and damages midbrain dopaminergic neurons. Hence, Rotenone toxicity doesnt solely result from a bioenergetics effect, but results primarily from oxidative damage (Sherer et al., 2003). This might be specially hazardous in children as they dont have a very well developed blood brain barrier and this would result in increased exposure/diffusion of Rotenone. The huge dosage of Rotenone produces much more pronounced effects as compared to smaller doses. Once exposed in childhood, or in younger adults, this produces much more amounts of ROS, and because of younger age, a much huger probability of developing undesirable genetic mutations later on in life.

HLTH 340: Environmental Toxicology and environmental health

Assignment 1

Bibliography: Atkinson, J. Arthur. Principles of clinical pharmacology 2007. Books.google.ca 27 February, 2011. Agent Orange Association of Canda. All you ever wanted to know about Dioxin or Perhaps you really do not want to know?.agentorangecanada.com <http://www.agentorangecanada.com/dioxin.php> 26 February, 2011. C. L., Olin, H. B. D., Dalhoff, K., & Jacobsen, J. (2006). In vivo human buccal permeability of nicotine. International Journal of Pharmaceutics, 311(1-2), 196-202. Ivey, K., & Triggs, E. (1978). Absorption of nicotine by the human stomach and its effect on gastric ion fluxes and potential difference. The American Journal of Digestive Diseases, 23(9), 809-814. doi:10.1007/BF01079790 Johnson and Johnson Inc. FAQs>What is Imodium 22 April, 2010. Imodium.ca < http://www.imodium.ca/en/faqs_aboutproducts_1.asp#question> 26 February, 2011. Kyerematen, G. A., & Vesell, E. S. (1991). Metabolism of nicotine. Drug Metabolism Reviews, 23(1-2), 3-41. doi:10.3109/03602539109029754 Sherer, et al. Machanism of Toxicity in Rotenone models of Parkinsons Disease 26 November, 2003. The Journal of Neuroscience, 26 November 2003, 23(34). Meeker-Connell, Ann. "How Nicotine Works" 02 January 2001. HowStuffWorks.com. <http://health.howstuffworks.com/wellness/drugs-alcohol/nicotine.htm> 26 February 2011. The Cancer Council. Psychoactive effects of nicotine, 2001. Tobaccoinaustralia.org. < http://www.tobaccoinaustralia.org.au/chapter-6-addiction/6-3psychoactive-effects-of-nicotine> 26 February 2011. World Health Organization. Dioxins and their effects on human health May 2010. Who.int < http://www.who.int/mediacentre/factsheets/fs225/en/index.html> February 26, 2011.