Autacoid Pharmacology

Reading assignment: Veterinary Pharmacology and Therapeutics, Ed. Richard Adam, 2001, 8th Edition, pp 403-438
Dr. C.S. Venugopal, Rm 2421 SVM Bldg Tele: (225) 578-9748 E-mail: cvenugopal@vetmed.lsu.edu

Autacoids
Learners objectives
1. Define autacoids, list important autacoids 2. Study at least two important functions of each one. 3. Therapeutic uses of autacoids and their antagonists 4. Learn mechanism of action of NO and endothelin 5. Vascular and nonvascular actions of leukotriens and PAF 6. Learn the importance of antagonists of histamine and serotonin and angiotensin

AUTACOIDS
Introduction

Means self remedy Naturally occurring substances Localized in tissues Do not normally circulate Diverse physiological and pharmacological activities Differ from hormones and neurotransmitters Short duration of action Usually involved in a response to injury Sites of action restricted to the synthesis area

CLASSIFICATION
1. Biogenic amines: Histamine, 5-hydroxytryptamine
2. Biogenic Peptides: Angiotensin and kinins 3. Small Proteins: cytokineins 4. Membrane derived lipids: LTs, PGs, TxA2 & PAF 5. Endothelium-derived agents:NO (gas); ET (peptide)

BIOGENIC AMINES
Histamine

Biosynthesis: Decarboxylation of histidine Source: Plants & animals (in all tissues) Storage:High in skin/epidermis, GI mucosa, (non-mast cells) and lungs (mast cells). Bound to heparin in mast cells. Basophils in blood Release: antigen antibody reaction leading to degranulation of cells (mast)

BIOGENIC AMINES
Histamine

Effects: allergic response, exocrine secretion (gastric acid,

pulmonary, lacrimal, salivary etc.), neurotransmission in CNS. Non-vascular smooth muscles contracted. Stimulates adrenal medulla (release epi); dilates cerebral vessels (histamine headache); capillary leak leads to hypotension/edema/histamine shock.

Receptors: H1(contractile), coupled with phospholipase;

H2(gastric secretion, vessel relaxation), cAMP coupled; H3(feed-back inhibition). Interact with G protein in the plasma membrane. In general, histamine constricts large vessels; dilates capillaries and venules

Biogenic amines
Histamine

Triple response: red spot (capillary

dilation); wheal (edema fluid); red flare (arteriole dilation-axon reflex) Fate: Gut absorption poor; metabolism by N-methyl transferase & histaminase Symptoms: anaphylaxis, swelling (skin, mucosa); itching, bronchospasm, hypotension, shock, phospholipase C and A2 activation.

Biogenic Amines
Histamine

Liberators: large molecules (proteins

egg white, serum, venom, toxins); surface active agents, proteolytic enzymes, drugs etc.

Clinical uses: diagnosis of achlorhydria,

diagnosis of pheochromocytoma, and to verify integrity of axon reflexes. Antagonists are used clinically.

ANTAGONISTS
of Histamine

H1 antagonists

Competitive antagonism for the same receptor do not prevent histamine release do not cause physiological antagonism (smooth muscle dilatation) Do not block gastric acid production (blocked by H2) Protect against shock and allergic reactions CNS depression, a side effect. Local anesthetic effect Therapeutic uses (in allergic reactions, hay fever, bronchial asthma, motion sickness, common cold, founder, insect bites Eg: Benadryl, Pyrilamine, etc.)

ANTAGONISTS
Of Histamine

H2 Antagonists Effect: Block all phases of gastric acid

secretion due to histamine. Uses: treatment of duodenal ulcer, stress related gastritis & peptic ulcer Agents: Burimamide, Metiamide, Cimetidine (Tagamet) Ranitidine (Zantac), Famotidine (Pepcid)

BIOGENIC AMINES
SEROTONIN

Source: plants (banana, pineapple, plums) &

animals (mollusks, arthropods, mammals (platelets, not in mast cells). Biosynthesis: Hydroxylation of tryptophan, then decarboxylation to serotonin(5-hydroxy tryptamine;5-HT). Rapidly absorbed into secretory granules. Accumulated in platelets, degredation by oxidative deaminaion. Uses: No therapeutic use. Antagonists are highly useful.

BIOGENIC AMINES
SEROTONIN

Actions

Neurotransmitter in the CNS Precursor of melatonin Induces sleep, Intestinal motility Involved in Temperature regulation Affects mood and behavior (humans) Deficiency causes depression Involved in hemostasis (platelets), Carcinoid syndrome (tumor of serotonin producing cells)

BIOGENIC AMINES
SEROTONIN

Receptors
5-HT1: vasodilatation through nitric oxide,
positive inotropic and chronotropic effects, increases small intestinal motility, inhibits stomach and large intestine

5-HT2: vasoconstriction 5-HT3: neuronal receptors influence uptake of

5-HT

ANTAGONISTS
OF SEROTONIN

Cyproheptadine: Specific for 5-HT2, & less specific for 5-HT1, has some H1 antagonism, weak Ca blocker. Katanserin: specific for 5-HT2, has affinity toward alpha-1, H1 and dopamine receptors.
Methysergide: Mixed 5-HT1 and 5-HT2 Fluoxetine (Prozac, Paxil, Zoloft): 5-HT3, neuronal uptake inhibitor

BIOGENIC PEPTIDES
ANGIOTENSIN

Synthesis & metabolism

Angiotensinogen (plasma substrate) to angiotensin I, by renin that is released during renal ischemia. Angiotensin I to II by angiotensin converting enzyme (ACE, Kinase II) in the pulmonary endothelium. Renin inhibited by propranolol Angiotensin II metabolized by angiotensinases in plasma and tissues.

BIOGENIC PEPTIDES
ANGIOTENSIN

Effects
Causes profound vasoconstriction

Increases peripheral vascular resistance Increases blood pressure Directly stimulates heart Facilitates epinephrine and aldosterone release Increases Na reabsorption in kidney tubules Releases ADH (vasopressin) to restore blood volume Saralasin blocks angiotensin (AT1 & AT2) receptors Captopril & enalapril inhibit ACE (congest. heart fail.).

BIOGENIC PEPTIDES
BRADYKININ

Actions
Comes under the group kinins This mediates nociception (pain) Regulates BP (vasodilator) Increases capillary permeability Balances electrolytes and fluid, Contracts gut slowly and stimulates prostaglandin synthesis Produced by tissue damage, viral infection, allergic reaction & inflammation Contracts various other smooth muscles

BIOGENIC PEPTIDE
BRADYKININ

Synthesis
Prekallikrein
(Plasma) Hagman factor (activated by collagen)

Kallikrein
(plasma & tissues)

1. Kininogen (alpha-2 globulins High and low m.w.

Bradykinin, Kallidin
Kallikrein (nonapeptide)(decapeptide) (Activated by peptidases)

Kallidin, a decapeptide, with same actions is also produced along with bradykinin. Both are metabolized to inactive agents by kinases II and ACE

BIOGENIC PEPTIDES
BRADYKININ & KALLIDIN

Receptors

B1 mediates vasoconstriction, sensitive to metabolites. B2 mediates vasodilatation, permeability, smooth muscle contraction, pain B3 mediates guinea pig tracheal contraction, not antagonized by B1 or B2 antagonists.

SMALL PROTEINS
CYTOKINES (TNFa, IL-1,IL-6)

1. TNF (Tumor Necrosis Factor ) A mediator of endotoxic shock Released by macrophages when exposed to endotoxin Triggers wide endotoxemic symptoms Elicits production of other cytokines, eicosanoids & humoral factors Stimulates degradation and adherence of neutrophils to endothelium Symptoms of septicemia at high concentration Can be neutralized by TNF antiserum Chronic exposure leads to cachexia (TNF was formerly known as cachectin)

SMALL PROTEINS
CYTOKINES

2. Interleukin-1 (IL-1) Known as lymphocyte activating factor ( T-cell responses) Activates endogenous pyrogen (induces fever) Interacts synergestically with TNF synthesis & release of IL-2 by interacting with antigen stimulated T-cells Activates B-cells and antibody synthesis arachidonic acid metabolism, inflammatory proteins neutrophil chemoattraction & fibroblast proliferation PGI2 synthesis in endothelial cells

SMALL PROTEINS
CYTOKINES

3. Interleukin-6 (IL-6)

A phosphoglycoprotein Produced and secreted by macrophates, monocytes, fibroblasts Inflammatory stimuli causes production by endothelium, T lymphocytes, mast cells IL-6 induces production of hepatic fibrinogen for protection against microorganisms Endotoxin, TNF alpha and IL-1 increase the levels of IL-6 Does not cause tissue injury or vascular thrombosis

MEMBRANE DERIVED AGENTS

EICOSANOIDS Derived from 20 carbon essential fatty acids Arachidonic acid (AA) is a 20 carbon essential fatty acid AA is the abundant precursor of eicosanoids in man AA is esterified to phospholipids of the cell membranes Biosynthesis depends on synthesizing enzyme availability Synthesis is regulated and initiated by physical & chemical stimuli These stimuli causes release of AA from the cellular stores

MEMBRANE DERIVED AGENTS

EICOSANOIDS

Biosynthesis

Physical & chemical stimuli interact with membranebound G protein receptors This interaction activates phopholipase C and/or phospholipase A2 which increases concentration of intracellular Ca Phospholipase A2 hydrolizes ester bond of phopholipids (AA) release Phodpholipase cleaves the bond and forms diglyceride Diglyceride lipase releases diglyceride arachidonate from diglycerides The released arachidonate is oxygenated by several enzymes Major enzymes are cyclooxygenase and lipoxygenase

EICOSANOIDS
Cyclooxygenase pathway
Arachidonic Acid Cyclooxygenase

PGG
Prostacyclin synthetase Thromboxane synthetase PGI2 PGH2 PGE2 PGD2 PGF2

TXA2

EICOSANOIDS
LIPOXYGENASE PATHWAY

Arachidonic Acid 12-lipoxygenase

12HPETE 5-HETE 12-HETE

5-lipoxygenase
5-HPETE LTA synthetase LTA4 Glutathione S

LTA hydrolase transferase LTB4 LTC4 Gamma glutamyl transpeptidase LTD4 Dipeptidase LTE4 Gamma glutanyl transpeptidase LTF4

EICOSANOIDS
Pharmacological actions

AA metabolites are present in all tissues & body fluids They have broad spectrum of biological activities In general, PGEs and PGIs are potent vasodilators PGF alpha and TXA are vasoconstrictors LTC and LTD capillary permeability & BP PGI inhibits platelet aggregation TXA induces platelet aggregation (aspirin inhibits) LTB is a potent chemotactic agent Bronchial muscles are relaxed by PGEs and PGIs PGFs, PGDs, LTC and LTD constrict bronchi
2 2 4 4 2

MEMBRANE-DERIVED AGENTS
Platelet Activating Factor (PAF)
General Information

Hanson (1971) identified a platelet aggregating agent Same substance (BP) was found in adrenal medulla Phospholipase A2 releases AA & lysoPAF from membr LysoPAF is acetylated by acetyl coenzyme A to PAF (catalyzed by acetyl transferase) Antigen-antibody reaction, chemotactic peptides, thrombin, collagen and other autacoids synthesis. regulated by Ca++ PAF acetyl hydrolase deacetylates for inactivation Platelets, neutrophils, monocytes, mast cells, eosinophils, renal medullary cells and endothelium synthesize PAF

PLATELET ACTIVATING FACTOR

Physiological functions

Involved in ovulation, implantation, parturition (antagonists are contraseptives) PAF in amniotic fluid derived from fetal lung. It is a potent bronchoconstrictor (long lasting) & causes pulmonary edema Simulates G proteins that are on the cell surface This activates phospholipase C & A2 Forms inositol phosphate, diacylglycerol and arachidonates Thus, PAF leads to the formation of PGs, LT and TXA2

PAF
Pharmacological actions

Increases platelet aggregation Potent vasodilator, PVR and BP Constricts pulmonary vessels Microvascular permeability Releases eicosanoids, generates superoxides Contracts nonvascular smooth muscles, Increases respiratory secretions, forms

pulmonary edema Decreases renal flow

ENDOTHELIUM-DERIVED AGENTS
Nitric Oxide (NO) A lipid soluble gas, slowly reactive radical, short half-life NOS deaminates L-arginine to L-citrulline & NO NO Breaks down to nitrates and nitrites Known as endothelium derived relaxing factor (EDRF) Organic nitrates (amyl nitrate, nitroglycerin, nitroprusside) release NO Nitric Oxide Synthase (NOS) antagonists counteracts vascular relaxation

ENDOTHELIUM-DERIVED AGENTS NITRIC OXIDE

Nitric Oxide Synthase (NOS)

NOS-1 (nNOS, bNOS): constitutive, low output form NOS-2 (MacNOS, iNOS): inducible, high output immune/inflammatory isoform that is found principally in monocytes and macrophages NOS-3 (eNOS): constitutive, low output form, vascular tone Control of constitutive forms of NOS (nNOS & eNOS) is mediated by intracellular calcium and calmodulin. Pulsatile flow and shear stress stimulate eNOS synthesis iNOS can be induced in macrophages, Kupffer cells, neutrophils, fibroblasts, vascular smooth muscle & endothelial cells.

ENDOTHELIUM-DERIVED AGENTS
EFFECTS OF NO

Physiological vasodilator, influences vascular

resistance and BP Homeostasis of vascular beds(cerebral, pulmonary, coronary) eNOS blockers increase BP by vasoconstriction Basal NO production in these beds regulates vascular tone NO mediates NANC neurotransmission It causes: relaxation of stomach & internal anal sphincter, and involved in penile erection

ENDOTHELIUM DERIVED AGENTS

EFFECTS OF NO

Induction of iNOS in neutrophils and monocytes,

increases NO Antiviral effect of interferon (IFN) is due to NO TNF and LT (pro-inflammatory agents) also induce NOS Macrophages kill microorganisms by NO Cytotoxic and/or cytostatic effects of non-specific host defense nNOS derived NO influences neuronal development, memory, vision and nociception

ENDOTHELIUM-DERIVED AGENTS Endothelin

Introduction

The most potent vasoconstrictor discovered to date First identified (1988) in porcine aortic endothelial cells Endothelin family has 3 isopeptides (ET-1, ET-2, ET-3) Each isopeptide is encoded by separate genes All ETs have 21 amino acids and 2 disulfide bonds Biologically & structurally ET is similar to sarafotoxin (snake venom) ET is a mitogen for vessels & myocardial cells ET has positive inotropic and antiarrhythmic effects ET can interact with inflammatory cytokines

ENDOTHELIUM-DERIVED AGENTS Endothelin

Biosynthesis

Preproendothelin (203 amino acids) is the precursor in plasma Converted by dibasic endopeptidase to big ET (39 amino acids) ET converting isoenzyme cleaves Big ET to active ET By cleaving, the C terminal is removed These enzymes are membrane bound metallopeptidases Metalloendopeptidases enzymatically degrade ET Norepi, angiotensin II, vasopressin, bradykinin, thrombin, LPS, TNF, and IL-1 stimulate ET precursor expression

ENDOTHELIUM-DERIVED AGENTS Endothelin Pharmacological effects

Type I reaction: (ET-1 and ET-2) vasoconstriction, bronchoconstriction, uterine contraction, stimulation of aldosterone synthesis Type II reaction: (ET-1, ET-2 & ET-3) all are equally effective in vasorelaxation & inhibition of platelet aggregation

ENDOTHELIUM-DERIVED AGENTS Endothelin

ET receptors

Two types have been identified (ETA and ETB) ETA is responsible for type I responses ETB is responsible for type II responses ETB1 produces vasodilatation by releasing NO and
PGI2 ETB2 produces nonvascular smooth muscle contraction.

ENDOTHELIUM-DERIVED AGENTS Endothelin

Physiological functions

ET is involved in the closure of the ductus

arteriousus Regulation of blood pressure Facilitates release of NO Responsible for tone of smooth muscles

ENDOTHELIUM-DERIVED AGENTS Endothelin

Diseases Hypertension laminitis associated with hypertension Cardiomyopathy & failure, pulmonary Hypertension Chronic obstructive pulmonary disease (COPD) Systemic Inflammatory Response Syndrome (SIRS) SIRS: sepsis, adult respiratory distress syndrome (ARDS), multiple organ failure, hemorrhagic shock, anaphylaxis High plasma ET in hypertrophy, cardiomyopathy, chronic heart failure, and asthma (BAL fluids) ET-1 probably mediates cardiac myopathy after hypoxia (ischemia) and pressure overload

ENDOTHELIUM-DERIVED AGENTS Endothelin Therapeutic uses Problems

Inhibitors of ET converting enzyme ET receptor antagonists ET is involved in complex reactions with

several other mediators ET has anti-arrhythmic effects Antagonists of ET have potential to predispose to arrhythmia These effects are under investigation now