Pcol322 Lab: Midterms - Post Lab 8: Antipyretic Activity PDF

PCOL322 LAB: Midterms POST LAB 8: ANTIPYRETIC ACTIVITY 4. After 60 mins. The test compound is administered either SQ or PO. Fever...

PCOL322 LAB: Midterms POST LAB 8: ANTIPYRETIC ACTIVITY 4. After 60 mins. The test compound is administered either SQ or PO. Fever 5. Body temperature is monitored for at least 3 hrs. - Condition in which there is a rise in body Aspirin temperature above the normal that is 37ºc. - Used to reduce fever and relieve mild to moderate - Usually due to bacterial or viral infections and can pain from conditions such as muscle aches, accompany any infectious disease. toothaches, common cold, and headaches. - “pyrexia” - May also be used to reduce pain and swelling in - Shivering, headache, nausea, and constipation or conditions such as arthritis. diarrhea accompanies it. - Elevations in body temperature occur when Felix Hoffman concentrations of prostaglandin E(2) (PGE(2)) increase within certain areas of the brain. These - Working at the Bayer company in Germany, made elevations alter the firing rate of neurons that control the acetylated form of salicylic acid in 1897. This thermoregulation in the hypothalamus. drug was named "Aspirin" and became the most - Although fever benefits the nonspecific immune widely used medicine of all time. response to invading microorganisms, it is also John Vane, 1971 viewed as a source of discomfort and is commonly suppressed with antipyretic medication. - Discovered the mechanism by which aspirin exerts its anti-inflammatory, analgesic and antipyretic Antipyretics actions. He proved that aspirin and other non- - Drug that prevent or reduce fever by lowering the steroid anti-inflammatory drugs (nsaids) inhibit the body temperature. activity of the enzyme now called cyclooxygenase - Thermoregulatory center of the brain is the one (COX) which leads to the formation of likely to be affected by these drugs, the prostaglandins (pgs) that cause inflammation, hypothalamus. swelling, pain and fever. - It is now clear that most antipyretics work by Paracetamol inhibiting the enzyme cyclooxygenase and reducing the levels of PGE(2) within the hypothalamus. - Commonly used medicine that can help treat pain - Paracetamol, Aspirin are some of the common and reduce a high temperature (fever). examples of antipyretics. - “Acetaminophen”acetyl-para-amino-phenol - Typically used to relieve mild or moderate pain, Screening Methods for Antipyretic Agents such as headaches, toothache or sprains, and - Antipyretic testing in rats reduce fevers caused by illnesses such as colds - Antipyretic testing in rabbits and flu. - MOA: Raising the threshold of the pain center in the Antipyretic Testing in Rats brain and by obstructing impulses at the pain- mediating chemoreceptors. The drug produces 1. A 15% suspension of brewer’s yeast in 0.9% NSS is antipyresis by an action on the hypothalamus; heat prepared. dissipation is increased as a result of vasodilation 2. Groups of six male or female wistar rats 150g are and increased peripheral blood flow. used. 3. Initial rectal temperatures are recorded. Telethermometer 4. The test animals are fevered by injection of 10ml/kg of brewer’s yeast suspension SQ in the back below - An apparatus for indicating the temperature of a the nape of the neck. distant point. 5. The site of injection in massaged in order to spread Brewer’s Yeast the suspension beneath the skin. The room is kept at 22-24 deg Celsius. - Derived from the yeast species Saccharomyces 6. Immediately after the yeast administration, food is cerevisiae (S. Cerevisiae), is commonly used for withdrawn. 18h post challenge, the rise in rectal inducing pyrexia in pharmacological studies temperature is recorded. screening antipyretics in rats. 7. The measurement is repeated after 30 min. Only - Subcutaneous administration of S. Cerevisae animals with body temperature of at least 38 deg induces prolonged fever, anorexia and lethargy that Celcius are taken into the test. is accompanied by a pronounced increase in the 8. The animals receive the test compound or the synthesis of pro-inflammatory cytokines, key standard drug by oral administration. Rectal prostaglandin synthesizing enzymes and temperatures are recorded again, 30, 60, 120, 180 transcription factors, in the periphery and brain. min. Post dosing. Antipyretic Testing in Rabbits 1. Rabbits 3-5 kg 2. Acclimatize in suitable cages for 60 min. 3. 0.2ml/kg containing 0.2 mcg LPS are injected IV into the rabbits ear. PCOL322 LAB: Midterms POST LAB 9: ANALGESIC ACTIVITY Screening methods for Centrally Acting Analgesics Hot Plate Method Pain - The paws of mice and rats are very sensitive to - Has been described by International Association for heat even at temperatures which do not damage the study of Pain as an “unpleasant sensory and the skin. emotional expressions associated with actual or - Response: jumping withdrawal of paws and licking potential tissue damage, or described in terms of of paws. such damage - 55-56 deg Celsius - First treated with non – narcotic analgesic. Haffner’s Tail Clip Test - Raised tail phenomena in mice are observed. - A clip is applied to the base of the tail of mice, and the reaction time is noted. - The animal respond quickly to the stimuli by biting the clip or the tail near the location of the clip. The time between the stimulation onset and response is measured by a stopwatch. - A metal artery clip was applied to the root of the mouse’s tail for 30 secs. Tail Immersion Test or Tail-Flick - This method is based on the observation that morphine-like drugs are selectively capable of prolonging the reaction time of the typical tail Pain Tolerance withdrawal reflex in rats induced by immersing the lower end of 5cm portion of the tail in warm water of - Is the maximum level of pain that a person is able to 55 deg Celsius. tolerate. Radiant Heat Method Analgesic - This test is useful for quantitative measurement of - Substance used to relieve pain. pain threshold against thermal radiation in man and - Aspirin (Aspilet) for evaluation of analgesic activity. - Acetaminophen (Biogesic) - It is very useful for discriminating between centrally - Indomethacin (Vi-Gel) acting morphine like analgesics and non opiate - Naproxen (Flanax) analgesics. ➔ Are some of the examples of analgesic agent. Formalin Test Method Screening Methods for Peripherally Acting Analgesic Agents - Rats weighing 180-300g are administered 0.05ml of 10% formalin solution into the lower surface of the Randall-Selitto Test (Pain in inflammed tissue) front paw. - Pain responses or excessive licking and biting of - This method is based on the principle that the paw. inflammation increases the peripheral analgesic sensitivity to pain. Tooth Pulp Stimulation - Inflammation decreases the pain reaction threshold, but the threshold is readily elevated by non-narcotic - This test is based on the fact that stimulation of the analgesic of the salicylate-amidopyrine type as well tooth pulp induces characteristic reactions, such as as by the narcotic analgesics. licking, biting, chewing, and head flick which can be observed easily. Writhing Test - Adult healthy rabbits are used for the test. Rabbits are anesthetized and their pulp chambers exposed - Pain is induced by injection of irritants into with a high speed chamber drills. On the day of the peritoneal cavity of the mice experiment, clamping electrodes are placed into the - The animals react with a characteristic stretching drilled holes. After an accommodation of 30 min. behavior which is called writhing. Stimulation is started to determine the threshold - Irritants: value the stimulus is applied with a frequency of 50 ➔ Phenyl quinone Hz and duration of 1 sec. The electrical current is ➔ Acetic acid started at 0.2 Ma and increase until the ➔ Irritants are injected Intraperitoneally to mice phenomenon of licking occurs. The test substance is either injected IV or given PO. PCOL322 LAB: Midterms Grid Shock Test - They can be used as monotherapy; however, they have greater efficacy in combination with weak and - This test measures the analgesic properties by the strong opioids. ‘flinch-jump’ procedures in rats. - Unlike opioids, long-term use of non-steroidal anti- - The floor of the box is wired with stainless steel inflammatory drugs does not lead to physical wires, spaced about 1 mm apart. The stimulus is dependence. given in the form of an electric current, 30 cycles Acetaminophen (paracetamol) per second with duration of 2 ms per pulse, with Anticonvulsants (including gabapentin and increasing shock intensities, the mice flinch, exhibit pregabalin) - nerve pain a starting reaction, increased locomotion or attempt Antidepressants (including amitriptyline and to jump. duloxetine) - nerve pain, muscle pain - The behavior is accurately reflected on the Aspirin (acetylsalicylic acid) oscilloscope by marked fluctuations of the pulse Other nsaids (including ibuprofen, diclofenac, and defined as the pained threshold response. The naproxen, and COX-2 inhibitors) current as a measured in milliampere is recorded Topical agents (including lidocaine and for each animal before and after administration of capsaicin) – numbness the drug. Electrical Stimulation DRUG MECHANISM OF EFFECT/S INTERACTION ACTION - This method is based on the fact that since the tail of the mice is known to be sensitive to any stimulus, the stimulus can be varied either by the duration of the electric current to check the efficacy of the Aspirin's Significant analgesic agent. antiplatelet Anticoagulants increase (up to properties may Percentage Inhibition (Warfarin and 1.5- to 2-fold) increase Heparin) in the risk of anticoagulation bleeding effect of warfarin. Aspirin increases effects of Increase glimepiride by Sulfonylureas tendency of plasma protein bleeding binding competition. Aspirin increases May increase levels of the effects of Methotrexate methotrexate by methotrexate decreasing renal in your body. clearance. Narcotic Analgesics Methotrexate - Describes a group of medications of the opioid class which have painkilling qualities. - May cause nausea, vomiting, diarrhea, sore throat, - Usually administered for the relief of severe pain chills, fever, rash, unusual bruising or bleeding, pale associated with fractures, burns, renal colic, skin, dark urine, swelling of the extremities, or coronary occlusion, etc. shortness of breath. - Ideally when you are given these medications they control (make tolerable) or at best stop the pain. - Because some narcotics are used recreationally or illegally some people associate the term narcotic with these uses. - Morphine, heroin, oxycodone, oxycontin, endone and codeine are narcotic analgesics Non-narcotic Analgesics - Commonly used to treat mild and moderate acute and chronic pain. PCOL322 LAB: Midterms POST LAB 10: HYPOGLYCEMIC Random blood sugar PROPERTY - Measure blood glucose regardless of when you last ate. Endocrine pancreas - Several random measurements may be taken throughout the day. - Consists of approximately 1 million islets of - Also called as “casual blood glucose test” langerhans interspersed throughout the pancreatic - Glucose levels in healthy people do not vary widely gland. throughout the day - Hormones secreted by this gland: - Blood glucose levels that vary widely may mean a Insulin problem → storage and anabolic hormone of the body Types of diabetes mellitus Islet amyloid polypeptide (iapp)/amylin → modulates appetite, gastric emptying, and Type 1 glucagon and insulin secretion - The hallmark of t1dm is selective beta cell (b cell) destruction and severe or absolute insulin Glucagon deficiency. → hyperglycemic factor that metabolizes glycogen stores Symptoms: Somatostatin - Feeling very thirsty → universal inhibitor of secretory cells - Passing urine more frequently, especially at night - Fatigue Pancreatic peptide - Weight loss and loss of muscle bulk → small protein that facilitates digestive - Thrush (persistent infection) processes by a mechanism not yet clarified Intervention: Ghrelin - Insulin replacement therapy is necessary to sustain → peptide known to increase pituitary growth life. hormone release - Pharmacologic insulin is administered by injection Hyperglycemia into the subcutaneous tissue using a manual injection device or insulin pump that continuously - This condition happens when there’s too much infuses insulin under the skin. sugar (glucose) in your blood. - Interruption can be life threatening and can result in - Happens when your body has too little insulin or if diabetic ketoacidosis or death your body can’t use insulin properly. - Insulin and glucagon are responsible for the Diabetec ketoacidosis is caused by insufficient or absent maintenance of blood sugar level. Imbalance of insulin and results from excess release of fatty acids and these two hormones results to hypo or subsequent formation of toxic levels of ketoacidosis. hyperglycemia. Type 2 Fasting blood sugar - Tissue resistance to the action of insulin combined - A test that measures sugar (glucose) in your blood. with a relative deficiency in insulin secretion It’s a simple, safe and common way to diagnose Symptoms: prediabetes, diabetes or gestational diabetes. - Don’t eat or drink anything for 8 to 12 hours before - Slow to heal cuts or ulcers the test. - Frequently vaginal or penile thrush - A healthcare provider will prick your finger or use a - Blurred vision needle to draw blood from the vein in your arm - Same as t1dm Intervention: Normal fasting blood sugar level 126mg/dl called nonketotic hypersmolar coma. main sign of diabetes - PCOL322 LAB: Midterms Nonketotic hypersmolar coma - the blood glucose may Metformin rise to 6-20 times the normal range and an altered mental state develops or the person loses consciousness. - Useful in prevention of t2dm - First-line therapy for t2dm Gestational SM - MOA: - Any abnormality in glucose level noted for the first ➔ A full explanation of the mecghanism od action time during pregnancy. of the biguanides remains elusive, but their - Diagnosed in app. 7% of all pregnancies in the usa primary effect is to activate the enzyme amp- - During pregnancy, the placenta and placental activated protein kinase (ampk) and reduce hormone create an insulin resistance that is most hepatic glucose production. pronounced in the last trimester. - Risk assessment for diabetes is suggested starting - Because metformin is an insulin-sparing agent and at the first prenatal visit. does not increase body weight or provoke - High-risk women should be screened immediately hypoglycemia, it offers obvious advanages over - Screening may be deferred in lower-risk women insulin or sulfonylureas in treating herglycemia in until the 24th and 28th week of gestation such persons. - Has a half life of 1.5-3hrs, is not bound to plasma Drugs used for diabetes proteins, is not metabolized, and is excereted by the - Insulins kidneys as the active compound. ➔ Rapid-acting: lispro, aspart, glulisine ➔ Short-acting: regular - Toxicity: gastrointestinal anorexia, nausea, ➔ Intermediate-acting: nph (neutral protamine vomiting, abdominal discomfort, and diarrhea; which hagedorn, or isophane) occur in up to 20% of patients. ➔ Long-acting: detemir, glargine - Sulfonylureas ➔ Glimepiride ➔ Gliclazide - Meglinitide analogs; α-phenylanaline derivative ➔ Mitiglinide - Biguanides ➔ Metformin - Alpha-glucosidase inhibitors ➔ Acarbose ➔ Miglitol - Thiazolidiones ➔ Pioglitazone - Glucagon-like polypeptide receptor agonists ➔ Exanatide ➔ Dulaglutide - Dipeptidyl peptidase-4 (dpp-4) inhibitors ➔ Sitagliptin ➔ Vildagliptin - Sodium-glucose co-transporter 2 (sglt2) inhibitors ➔ Dapagliflozin ➔ Empagliflozin - Islet amyloid polypeptide analog ➔ Pramlintide - Bile acid sequestrant ➔ Colesevelam hydrochloride - Dopamine agonist ➔ Bromocriptine PCOL322 LAB: Midterms POST LAB 11: ANTISPASMODIC AGENT Antispasmodics - Have been used for decades as an empirical Spasticity treatment of IBS - may be defined as “disordered sensorimotor control ➔ Irritable bowel syndrome (IBS) is a group of resulting froman upper motor neuron lesion, symptoms that occur together, including presenting an intermittent or sustained involuntary repeated pain in your abdomen and changes in activation of muscles.” your bowel movements, which may be - it is characterized by an increase in tonic stretch diarrhea, constipation, or both. reflexes and flexor muscle spasms together with - Examples: muscle weakness Mebeverine - often associated with spinal injury, cerebral palsy, Trimebutine multiple sclerosis and stroke; often involve Otilonium abnormal function of the bowel and bladder as well Hyoscine bromide – commonly used as skeletal muscle Cimetropium bromide - the mechanisms underlying clinical spasticity Dicyclomine hydrochloride – commonly used appear to involve not only the stretch reflex arc itself Hyoscyamine but also higher centers of the CNS, with damage to Dicyclomine descending pathways in the spinal cord resulting in Charcoal Meal Test hyperexcitability of the alpha motor neurons in the cord. - Method used to study the effect of drugs on the motility of intestine. Atropine - Acts as a competitive, reversible antagonist of muscarinic receptors: anticholinergic drug - Sources: Deadly nightshade → Atropa belladonna Jimson weed/Jamestown weed → Datura stramonium - aka hyoscyamine - MOA: ➔ Blockade of muscarinic receptors has dramatic effects on motility and some of the secretory functions of the gut ➔ In general, antimuscarinic drugs diminish the tone and propulsive movements; the walls of the viscera are relaxed. ➔ however, even complete muscarinic block cannot abolish activty in this organ system, since local hormones and noncholinergic neurons in the enteric nervous system also moduloated gastrointestinal function. ➔ GI smooth muscle motility is affected from the stomach to the colon ➔ therefore, gastric emptying time is prolonged, and intestinal transmit time is lengthened Neostigmine - Parasympathomimetic, specifically, a reversible cholinesterase inhibitor - all cholinesterase inhibitors increase the conc of endogenous ACh at cholinoceptors by inhibiting acetylcholinesterase - MOA: acetylcholinesterase inihibitor can enhance gastric, small intestine, and colonic emptying cholinergic effects include excessive salivation, nausea, vomiting, diarrhea, and bradycardia PCOL322 LAB: Midterms POST LAB 12: DIURETIC PROPERTY Diuretic Agents Functions of Kidneys MECHANIS CLASS OF M OF EXAMPLE - “A WET BED” DIURETICS ACTION A- maintaining ACID-base balance W- maintaining WATER balance E- ELECTROLYTE balance Inhibition of T- TOXIN removal the enzyme B- BLOOD pressure control prevents E- production of ERYTHROPOIETIN dehydration CARBONIC D- vitamin D metabolism of H2CO3 ANHYDRASE Acetazolamide and hydration INHIBITORS - Glumerolar filtration is the first step in making of CO2 in the urine. It is the process that your kidneys use to filter proximal excess fluid and waste products out of the blood convoluted into the urine collecting tubules of the kidney, so tubule they may be eliminated from your body. - Making ERYTHROPOIETIN → hormone secreted Inhibition of by the kidneys that increases the rate of production sodium/gluco of red blood cells in response to falling levels of se oxygen in the tissues. SODIUM cotransporter - Vitamin D metabolism → control calcium and GLUCOSE (SGLT2) phosphorus levels in the blood COTRANSPORT Canagliflozin results in ER 2 (SGLT2) decreased Diuretics INHIBITORS Na+ and - Drug that increase the flow of urine gluvose - First line agents tried in mild hypertension and reabsorption affects blood sodium level and blood volume. - Also inhibits sodium reabsorption to prevent edema Inhibition of - Agents that increases urine volume the Na/K/2Cl - AKA Water pills LOOP transporter in Furosemide Natriuretic DIURETICS the ascending - Causes an increase in renal sodium excretion loop of henle Aquaretic Pharmacologi - Increases excretion of solute-free water c antagonist of Clinical Uses of Diuretics aldosterone in collecting Edematous State Non-Edematous State tubule; weak Spironolactone antagonism of androgen Congestive Heart Failure Hypertension POTASSIUM- receptors SPARING Kidney Disease Nephrolithiasis DIURETICS Hepatic Cirrhosis Hypercalcemia Amiloride Blocks epithelial Idiopathic Edema Diabetes Insipidus sodium channels in collecting tubule Physical osmotic effect on tissue water OSMOTIC distribution Mannitol DIURETICS because it is retained in the vascular compartment PCOL322 LAB: Midterms Antagonist at POST LAB 13: ANTI-INFLAMMATORY V1a and V2 AGENT ADH receptors Conivaptan Inflammation VASOPRESSIN - Characterized by the orderly occurrence of several (ADH) processes: initiation, recruitment and ANTAGONIST Selective Tolvaptan chemoattraction of inflammatory cells and activation antagonist at of these cells to release inflammatory mediators V2 ADH capable of damaging or killing an invading microbe receptor or tumor. Inhibition of Mechanism of Inflammation the Na/Cl Hydrochlorothiazi How Inflammation Occurs in the Body? transporter in de THIAZIDES the distal - The inflammatory response (inflammation) occurs convoluted Metolazone when tissues are injured by bacteria, trauma, toxins, tubule heat, or any other cause. The damaged cells release chemicals including histamine, bradykinin, and prostaglandins. These chemicals cause blood vessels to leak fluid into the tissues, causing Furosemide swelling. - Prostaglandins play a key role in the generation of - Loop diuretic the inflammatory response. Their biosynthesis is - Help treat fluid retention (edema) and swelling that significantly increased in inflamed tissue and they is caused by congestive heart failure, liver disease, contribute to the development of the cardinal signs kidney disease, or other medical conditions of acute inflammation. PGE2 and PGI2, are the - MOA: decreases the sodium, chloride and predominant prostanoids associated with potassium reabsorption from the tubule, therefore, inflammation. Both markedly enhance edema increasing the excretion od sodium and water by formation and leukocyte infiltration by promoting the kidneys. blood flow in the inflamed region. Foods rich in Potassium 5 Cardinal Signs of Inflammation FRUITS VEGETABLES LENTILS 1. Rubor (Redness) 2. Calor (Increased heat) 3. Tumor (Swelling) dried acorn kidney beans 4. Dolor (Pain) apricots aquash 5. Functio laesa (Loss of function) soybeans prunes potatoes Three Phases of Inflammation nuts raisins spinach 1. The first phase being oedema and swelling with accompanying pain orange tomatoes 2. Second phase, leukocytes migrate to this area and juice mopping up operation starts broccoli 3. Followed by repair, which is ushered by the bananas proliferation of fibroblasts and synthesis of connective tissue Carrageenan - Made from parts of various red seaweeds in the Rhodophyceae family. - Additive used to thicken, emulsify, and preserve foods and drinks. - inflammation induced by activation of TLR2/6 and TLR4/6 are the predominant pathways through which carrageenan induces inflammatory responses. PCOL322 LAB: Midterms Inflammation - Is now rarely used as an anti-inflammatory medication. CHRONIC - Epidemiologic studies suggest that long-term use of ACUTE INFLAMMATION aspirin at low dosage is associated with a lower INFLAMMATION incidence of colon cancer, possibly related to its COX-inhibiting effects. 1% Carrageenan Cotton Pellet Granuloma Anti-inflammatory Agents 75mcg Croton oil ear Complete Freund's - A type of medicine that treats swelling and redness. edema Adjuvant - Mediators that reduce the production or activities of proinflammatory cytokines and block immune cell 2% Oxazolone in ear trafficking into tissues, hence they may develop to treat inflammation. - Non-Steroidal Anti-Inflammatory Drugs UV radiation in ear (NSAIDs) Aspirin 75mcg Croton oil ear edema COX-2 selective inhibitors - Ear inflammation is produced by applying a 2% ➔ Celcoxib solution of croton oil on the ears of mice or rats. ➔ Meloxicam 2% Oxazolone in ear Non-selective COX inhibitors ➔ Diclofenac - application on inner and outer surfaces of ear ➔ Ibuprofen ➔ Indomethacin UV radiation in ear ➔ Ketoprofen - exposure to ultraviolet (UV) light ➔ Ketorolac ➔ Naproxen Cotton Pellet Granuloma ➔ Piroxicam - subcutaneous implantation of a cotton pellet into a - Disease-modifying Antirheumatic Drugs rodent results in the formation of a granuloma at the (DMARDs) site of the implant ➔ Azathioprine Complete Freund's Adjuvant ➔ Chloroquine/Hydroxychloroquine ➔ Cyclophosphamide - oil-based component that when injected diffuses ➔ Cyclosporine into the surrounding tissue causing an inflammatory ➔ Methotrexate reaction ➔ Mycophenolate mofetil ➔ Rituximab Plethysmometer ➔ Tocilizumab ➔ TNF-a blocking agents - Designed to provide a highly useful tool in the ➔ Adalimumab measurement of small volume changes. ➔ Etanercept - This test is typically used to follow the evolution of the inflammatory response experimentally induced in rodents and to screen potential anti-inflammatory or antiedema properties of pharmacological substances Principles - For screening of inflammation or edema in mouse or rat - Is a micro controlled volume meter - Specially designed foe accurate measurement of the rat/mouse paw swelling Aspirin - One of a group of drugs called non-steroidal anti- inflammatory drugs (NSAIDs) - Inhibit the activity of the enzyme now called cyclooxygenase (COX) which leads to the formation of prostaglandins (PGs) that cause inflammation, swelling, pain and fever.