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!

! aa are oxygenation products of polyunsaturated long-chainare oxygenation products of polyunsaturated long-chain fatty acids

fatty acids !

! are ubiquitous in the animal kingdom and are also found— are ubiquitous in the animal kingdom and are also found—  together with their precursors—in a variety of plants. together with their precursors—in a variety of plants.

!

! 5,8,11,14-eicosatetraenoic acid5,8,11,14-eicosatetraenoic acid !

! the most abundant of the eicosanoid precursorsthe most abundant of the eicosanoid precursors !

! is a 20-carbon (C20) fatty acid containing four doubleis a 20-carbon (C20) fatty acid containing four double bonds (designated C20:4–6).

bonds (designated C20:4–6). !

! must first be released or mobilized from the sn-2 positionmust first be released or mobilized from the sn-2 position of membrane phospholipids by one or more lipases of the of membrane phospholipids by one or more lipases of the phospholipase A2 for eicosanoid synthesis to occur phospholipase A2 for eicosanoid synthesis to occur 3 classes of phospholipases mediate arachidonate release from 3 classes of phospholipases mediate arachidonate release from membrane lipids:

membrane lipids: 1.

1. cytosolic (c) PLA2- dominates the acute release of AAcytosolic (c) PLA2- dominates the acute release of AA 2.

2. secretory (s) PLA2- contributes under conditions ofsecretory (s) PLA2- contributes under conditions of sustained or intense stimulation of AA production sustained or intense stimulation of AA production 3.

3. calcium-independent (i) PLA2.calcium-independent (i) PLA2. !!

4 routes that oxygenate the AA 4 routes that oxygenate the AA

1.

1. the cyclooxygenase (COX)the cyclooxygenase (COX) 2.

2. lipoxygenaselipoxygenase 3.

3. P450 epoxygenaseP450 epoxygenase 4.

4. isoeicosanoiisoeicosanoid d pathwayspathways

Factors determining the type of eicosanoid synthesized: Factors determining the type of eicosanoid synthesized:

1.

1. the substrate lipid speciesthe substrate lipid species 2.

2. the type of cellthe type of cell 3.

3. the manner in which the cell is stimulatedthe manner in which the cell is stimulated

!

! Two unique COX isozymes convert AA into prostaglandinTwo unique COX isozymes convert AA into prostaglandin endoperoxides: endoperoxides: 1. 1. PGH synthase-1PGH synthase-1 most cells. most cells. 2. 2. PGH synthase-2PGH synthase-2

varies depending on the stimulus. varies depending on the stimulus.

!!  An immediate early-respo An immediate early-respo nse gene produnse gene product that is ct that is markedly up-markedly up-regulated by shear stress, growth factors, tumor promoters, and regulated by shear stress, growth factors, tumor promoters, and cytokines.

cytokines.

the major source of the major source of prostanoids in prostanoids in inflammation and inflammation and cancer cancer

promote the uptake of promote the uptake of two molecules of two molecules of oxygen by cyclization oxygen by cyclization of arachidonic acid to of arachidonic acid to yield a C9–C11 yield a C9–C11 endoperoxide C15 endoperoxide C15 hydroperoxide hydroperoxide vascular vascular prostacyclin prostacyclin (PGI2) (PGI2) !! prostanoids are prostanoids are important for important for normal renal normal renal development development and and maintenance of maintenance of function function equipotent on COX-1 equipotent on COX-1 and COX-2 and COX-2

!! C9–C11 endoperoxide C15 hydroperoxide- rapidly modified byC9–C11 endoperoxide C15 hydroperoxide- rapidly modified by the peroxidase moiety of the COX enzyme to add a 15-hydroxyl the peroxidase moiety of the COX enzyme to add a 15-hydroxyl group that is essential for biologic activity

group that is essential for biologic activity !! TheThe

termed the termed the

action of downstream isomerases and synthases action of downstream isomerases and synthases !! The prostaglandins differ from each other in two ways:The prostaglandins differ from each other in two ways:

1.

1. in the substituents of the pentane ring (indicated by thein the substituents of the pentane ring (indicated by the last letter, eg, E and F in PGE and PGF)

last letter, eg, E and F in PGE and PGF) 2.

2. in the number of double bonds in the side chains (indicatedin the number of double bonds in the side chains (indicated by the subscript, eg, PGE1, PGE2).

by the subscript, eg, PGE1, PGE2). !! PGH2 is metabolized byPGH2 is metabolized by

synthases (PGIS, TXAS, and PGFS) to synthases (PGIS, TXAS, and PGFS) to respectively

respectively

EICOSANOIDS

EICOSANOIDS

ARACHIDONIC

ARACHIDONIC ACID ACID OTHER OTHER POLYUNSATURATEDPOLYUNSATURATED PRECURSORS

PRECURSORS

phospholipase C

phospholipase C and diglyceride lipaseand diglyceride lipase- can also release AA- can also release AA

SYNTHESIS OF

SYNTHESIS OF EICOSANOIDSEICOSANOIDS Products of Prostaglandin Endoperoxide Synthases Products of Prostaglandin Endoperoxide Synthases (Cyclooxygenases)

(Cyclooxygenases)

(COX-1)

(COX-1)is expressed constitutively inis expressed constitutively in (COX-2)

(COX-2)is inducible; its expressionis inducible; its expression

COX

COX 1 1 COX COX 2 2 BOTHBOTH

generates generates prostanoids for prostanoids for “housekeeping” such “housekeeping” such as gastric epithelial as gastric epithelial cytoprotection cytoprotection Indomethacin and Indomethacin and sulindac (slightly sulindac (slightly selective COX 1) selective COX 1) !! source of source of Meclofenamate and Meclofenamate and ibuprofen are ibuprofen are approximately approximately endothelial endothelial

COX- is the primary  is the primary

renal renal COX-2-derived derived

prostaglandins, thromboxane, and

prostaglandins, thromboxane, and prostacyclprostacyclinin, collectively, collectively prostanoids

prostanoids, are generated from, are generated fromPGH2PGH2 through the through the

prostacycl

prostacyclin, thromboxane, and in, thromboxane, and PGFPGF

PGI2, TXA2, and PGF2 PGI2, TXA2, and PGF2!!,,

(2)

! 9,11-endoperoxide reductase and 9-ketoreductase- 2 additional enzymes that provide for PGF2" synthesis from PGH2 and

PGE2, respectively. !  At least

1. microsomal (m) PGES-1

2. mPGES-2- the more readily inducible 3. cytosolic PGES- the more readily inducible ! two distinct PGDS isoforms

1. lipocalin-type PGDS 2. hematopoietic PGDS

Several products of the arachidonate series with clinical importance: DRUG CLINICAL INDICATIONS

 Alprostadil may be used for its smooth muscle relaxing effects to maintain the ductus arteriosus patent in some neonates awaiting cardiac surgery and in the treatment of impotence.

Misoprostol, cytoprotective prostaglandin used in preventing peptic ulcer and in combination with

mifepristone (RU-486) for terminating early pregnancies.

PGE2 used in obstetrics to induce labor

Latanoprost topically active PGF2" derivatives used in

ophthalmology to treat open-angle glaucoma. Prostacyclin (PGI2,

epoprostenol)

synthesized mainly by the vascular endothelium and is a powerful vasodilator and inhibitor of platelet aggregation. It is used clinically to treat pulmonary hypertension and portopulmonary hypertension.

!

of platelets, vasoconstriction ).

!  All the naturally occurring COX products undergo rapid metabolism to inactive products either by:

 A. B.

corresponding ketone by prostaglandin 15-hydroxy prostaglandin dehydrogenase (15-PGDH) after cellular uptake via an organic anion transporter polypeptide (OATP 2A1).

! Further metabolism is by , and"

!  AA (metabolized by

hydroperoxyeicosatetraenoic acids (HPETEs)" derivatives (HETEs) and leukotrienes

!

recognized as the primary components of the anaphylaxis.

! There are four current approaches to antileukotriene drug development:

1. 5-LOX enzyme inhibitors

2. leukotriene-receptor antagonists 3. inhibitors of FLAP

4. phospholipase A2 inhibitors.

!

appropriate stimulation via 12-LOX in platelets to the L

!

and ichthyosis

! Specific isozymes of microsomal cytochrome P450

monooxygenases convert AA to hydroxy- or epoxyeicosatrienoic acids

! The products are:

o 20-HETE- generated by the CYP hydroxylases (CYP3A,

4A, 4F)

o 5,6-, 8,9-, 11,12-, and 14,15-epoxyeicosatrienoic

acids (EETs) - arise from the CYP epoxygenase (2J, 2C)

! EETs- have vasodilator effect except on pulmonary vasculature where they cause vasoconstriction.

! are family of eicosanoid isomers, are formed

nonenzymatically by  on AA

and related lipid substrates. !

! have potent vasoconstrictor effects when infused into renal and other vascular beds and may activate prostanoid receptors

! may modulate other aspects of vascular function, including leukocyte and platelet adhesive interactions and

angiogenesis !

!

inhibition with aspirin or other NSAIDs should not affect the isoprostane pathway

! Peroxidation of arachidonate by free radicals- primary epimerization mechanism in the production of isoeicosanoids

! eicosanoids act mainly in an fashion

! PGI2 (IP), PGF2" (FP), and TXA2 (TP) receptors- has a single

gene product

! PGE2- have 4 receptors ! PGD2 – have 2 receptors

! LTB4 (BLT1 and BLT2) and the cysteinyl leukotrienes- both have 2 receptors

#

 All of these receptors are G protein-coupled

! have major effects on smooth muscle in the vasculature, airways, and gastrointestinal and reproductive tracts. three PGE2 synthases have been identified:

(PGE1)

a PGE1 derivative

andPGF2

Thromboxane(TXA2)- has undesirable properties (aggregation

hydration(for PGI2 and TXA2)

oxidation of the key 15-hydroxyl group to the

13 reduction,!-oxidation oxidation

Products of Lipoxygenase

5-, 12-, and 15-lipoxygenases LOX) hydroxy LTC4andLTD4are potent bronchoconstrictors and are

slow-reacting substance of anaphylaxis (SRS-A)that is secreted in asthma and

LTA4- the primary product of 5-LOX, can be converted with lipoxins XA4 andLXB4 in vitro

Epidermal accumulation of 12( )-HETE is a feature of psoriasis

Epoxygenase Products

Isoeicosanoids

direct free radical-based action

Isoprostanes are prostaglandin stereoisomers

COX is not needed for the formation of the isoprostanes, and its

BASIC PHARMACOLOGY OF EICOSANOIDS

MECHANISMS EFFECTS OF EICOSANOIDS

autocrine and a p aracrine

(3)

!

, while relaxing effects are mediated by the generation of cAMP.

SMOOTH MUSCLE

Contraction of smooth muscle is mediated by the release of calcium

VASCULAR SMOOTH MUSCLES PROSTAGLANDINS

potent vasoconstrictor smooth muscle cell and is the

has convincingly been shown to have this effect.

PGE2  PGI2 

PGD2 

*

testosterone, which up-regulates smooth muscle cell TP expression

!  Another vasoconstrictor is the

PROSTAGLANDINS PGE2 (  PGF2 ! PGE2  PGE2 or PGF2 ! PROSTAGLANDINS Contraction by TXA2

! - are also bronchodilators

! act principally on smooth muscle in peripheral airways and are a thousand times more potent than histamine, both in vitro and in vivo

! also stimulate bronchial mucus secretion and cause mucosal edema

PLATELET

! Low concentrations of PGE2 enhance platelet aggregation (via EP3), whereas higher concentrations inhibit (via IP) ! Both

! TXA2 is the major product of COX-1, the only COX isoform expressed in mature platelets. Itself a platelet aggregator, TXA2 amplifies the effects of other, more potent, platelet agonists such as thrombin.

! Platelet COX-1-derived TXA2 biosynthesis is increased during platelet activation and aggregation and is irreversibly inhibited by chronic administration of aspirin at low doses

KIDNEY

! medulla and the cortex of the kidney synthesize prostaglandins, the medulla substantially more than the cortex

! COX-1 is expressed mainly in cortical and medullary collecting ducts and mesangial cells, arteriolar endothelium, and epithelial cells of Bowman’s capsule.

! COX-2 is restricted to the renal medullary interstitial cells, the macula densa, and the cortical thick ascending limb. ! major renal eicosanoid products are

followed by PGF2" and TXA2

! also synthesizes several hydroxyeicosatetraenoic acids, leukotrienes, cytochrome P450 products, and epoxides !

pressure and regulating renal function, particularly in marginally functioning kidneys and volume-contracted states.

! renal cortical COX-2-derived PGE2 and PGI2 maintain renal blood flow and glomerular filtration rate through their local vasodilating effects

!

inhibit tubular sodium reabsorption, while products promote salt excretion in the collecting ducts.

! Loop diuretics, eg, furosemide, produce some of their effect by stimulating COX activity.

o patient response to a loop diuretic is diminished if a COX

inhibitor is administered concurrently

! TXA2 causes intrarenal vasoconstriction (and perhaps an ADH-like effect), resulting in a decline in renal function.

! Hypertension is associated with increased TXA2 and decreased PGE2 and PGI2 synthesis

o increased TXA2 formation has been reported in

nephrotoxicity TXA

vasoconstrictor (PGF2!

not a smooth muscle mitogen mitogen*

only eicosanoid that

Vasodilator (especially PGI2 and PGE2)- promote

vasodilation by increasing cAMP and decreasing smooth muscle intracellular calcium, primarily via the IP and EP4 receptors.

 is a vasodilator produced by endothelial cells.

 inhibits proliferation of smooth muscle cells, an action that may be particularly relevant in pulmonary hypertension.

 may also function as a vasodilator—in particular as a dominant mediator of flushing induced by the lipid-lowering drug niacin

mitogenic effect is potentiated by exposure of smooth muscle cells to

isoprostane 8-iso-PGF2!

SMOOTH MUSCLES of the GIT

TXA Most of the prostaglandins and thromboxanes activate gastrointestinal smooth muscle

via EP3) andPGF2 ! (via

FP)-contract the longitudinal smooth muscle  and weakly byPGI2-  contract the circular muscle

 (via EP4)- relaxes the circular smooth muscle

results in colicky cramps AIRWAY SMOOTH MUSCLES

TXA PGE2 and PGI2 RELAXATION

PGD2 and PGF2! CONTRACTION

cysteinyl leukotrienes

PGD2 and PGI2 inhibit aggregation

PGE2 and PGI2,

Prostaglandins  play important roles in maintaining blood

COX- 2-derived prostanoids  increase medullary blood flow and COX-1-derived

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REPRODUCTIVE ORGANS

(FEMALE)

! Uterine muscle is

! .

! PGF2", together with oxytocin, is essential for the onset of parturition.

REPRODUCTIVE ORGANS

(MALE)

!  A major source of these prostaglandins is the seminal vesicle; the prostate, despite the name “prostaglandin,” and the testes synthesize only small amounts.

! Smooth muscle-relaxing prostaglandins such as PGE1 enhance penile erection by relaxing the smooth muscle of the corpora cavernosa

CNS and PNS

1. FEVER

! PGE2 increases body temperature, predominantly via EP3

!

PGD2 and TXA2 do not

! Endogenous pyrogens release interleukin-1, which in turn promotes the synthesis and release of PGE2

! synthesis is blocked by aspirin and other antipyretic compounds.

2. SLEEP ! !

3. NEUROTRANSMISSION

! PGE compounds inhibit the release of norepinephrine from postganglionic sympathetic nerve endings

! Both COX-1 and COX-2 are expressed in the spinal cord and release prostaglandins in response to peripheral pain stimuli

INFLAMMATORY AND

IMMUNITY

! PGE2 and PGI2 are the predominant prostanoids associated with inflammation

• enhance edema formation and leukocyte infiltration

! PGE2 and PGI2, through activation of EP2 and IP, respectively, increase vascular permeability and leukocyte infiltration.

! TXA2 can also increase platelet-leukocyte interactions. ! PGE2 suppresses the immunologic response by inhibiting

differentiation of B lymphocytes into antibody-secreting plasma cells

! PGD2, a major product of mast cells, is a potent chemoattractant for eosinophils

• also induces chemotaxis and migration of TH2

lymphocytes

BONE METABOLISM

!

produced by osteoblasts and adjacent hematopoietic cells

o increase bone turnover

EYE

! PGE and PGF derivatives lower intraocular pressure

CANCER

! In familial polyposis coli, COX inhibitors significantly decrease polyp formation

!

prostanoid, facilitates tumor initiation, progression, and metastasis through multiple biologic effects, increasing proliferation and angiogenesis, inhibiting apoptosis, augmenting cellular invasiveness, and modulating immunosuppression

Blood Cells and Inflammation ! LTB4 ! cysteinyl leukotrienes  -! Heart and

Smooth Muscle vascular smooth muscle cell proliferation and migration at low concentrations

!  reduce myocardial

contractility and coronary blood flow, leading to cardiac depression. cells synthesize

neutrophils. The

bronchoconstrictors and cause increased micro- vascular permeability, plasma

exudation, and mucus secretion in the airways Renal System ! 20-HETE, which potently blocks the

smooth muscle cell Ca2+-activated K+ channel and leads to vasoconstriction of the renal arteries" HPN

contracted by PGF2#, TXA2, and low concentrations of PGE2

PGI2 and high concentrations of PGE2 causerelaxation

Exogenous PGF2#  and PGI2 induce fever, whereas

PGD2 induces natural sleep 

PGE2  infusion into the posterior hypothalamus causes wakefulness.

Prostaglandins  are abundant in skeletal tissue and are

PGE2, which is considered the principal oncogenic

Effects of Lipoxygenase Cytochrome

P450-Derived Metabolites

, acting at the BLT1 receptor, is a potentchemoattractant  for T

lymphocytes, eosinophils, monocytes,  are potent chemoattractants for eosinophils and T lymphocytes.

also contribute to inflammation by increasing endothelial permeability, thus promoting migration of

inflammatory cells to the site of inflammation

Bothlipoxin A and lipoxin B  inhibit natural killer cell cytotoxicity.

Cardiovascular—12( )- HETE promotes

LTC4 and LTD4

Gastrointestinal Human colonic epithelial LTB4, a chemoattractant for Airways— cysteinyl leukotrienes,

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!   block all the known pathways of eicosanoid synthesis.

- They inhibit phospholipase A2 activity, probably by interfering with phospholipid binding, thus preventing the release of arachidonic acid.

! NSAIDs (eg,

- block both prostaglandin and thromboxane formation by reversibly inhibiting COX activity.

!

! 5-LOX inhibitor (

receptor for leukotrienes ( and

FEMALE REPRODUCTIVE ORGANS

! PGE2 and PGF2" have potent oxytocic actions.

! by promoting uterine contractions

!

vaginally for oxytocic use.

! it is approved for

induction of labor in patients at or near term.

!  A stimulates the contraction of the uterus through- out

pregnancy.

! also directly affects the collagenase of the cervix,

resulting in softening.

• Excreted in urine • T" is 2.5- 5 mins.

• 20-mg dinoprostone vaginal suppository repeated at

3-to 5-hour intervals – recommended dose for abortifacient purposes

• mean time to abortion is 17 hours,

!  Antiprogestins (eg,

oral oxytocic synthetic analog of PGE1 ( early abortion.

!

! used to induce second-trimester abortions and to

control postpartum hemorrhage that is not responding to conventional methods of management.

SIDE EFFECTS:

! Prostaglandins have moderate AE, with a slightly higher incidence of nausea, vomiting, and diarrhea than that produced by oxytocin ! PGF2" has more gastrointestinal toxicity than PGE2

! PGF2" is a bronchoconstrictor and should be used with caution in women with asthma

! both PGE2 and PGF2" pass the fetoplacental barrier, fetal toxicity is uncommon

"

labor in women with preeclampsia-eclampsia or cardiac and renal

diseases because, unlike oxytocin, they have no antidiuretic effect.

Dysmenorrhea

!

in patients with primary dysmenorrhea !

potency and is quickly hydrolyzed,

MALE REPRODUCTIVE ORGANS

! (PGE1) is a second-line treatment for erectile dysfunction

o

related to the algesic effects of PGE derivatives

RENAL SYSTEM

! Bartter’s syndrome – results when synthesis of prostaglandins is increased.

! characterized by low-to-normal blood pressure,

decreased sensitivity to angiotensin, hyperreninemia, hyperaldosteronism, and excessive loss of K+

CARDIOVASCULAR SYSTEM

! !

vascular resistance ! PGI2 (

• side effects include flushing, headache,

hypotension, nausea, and diarrhea. !

! may be delivered by subcutaneous or intravenous infusion.

!

! use of PGE1 and PGI2 compounds in Raynaud’s phenomenon and peripheral arterial disease

!

! COX inhibitors are often used to inhibit synthesis of PGE2 and so close the ductus.

BLOOD

! Chronic administration of low-dose aspirin" selectively and irreversibly inhibits platelet COX-1 without modifying the activity of systemic COX-1 or COX-2

RESPIRATORY SYSTEM

!

form " promotes !

were once thought to be primary mediators in ! leukotriene-receptor inhibitors (eg,

are effective in asthma.

! Corticosteroids and cromolyn are also useful in asthma

INHIBITION OF EICOSANOID SYNTHESIS

Corticosteroids

indomethacin, ibuprofen)

Aspirin is an irreversible COX inhibitor.

zileuton) and selective antagonists of the CysLT1 zafirlukast, montelukast, pranlukast-used clinically in mild to moderate asthma.

CLINICALPHARMACOLOGYOF EICOSANOIDS

Dinoprostone,a synthetic preparation of PGE2, is administered inducing abortion in the second trimester of pregnancy, formissed abortion, for benign hydatidiform mole, and for ripening  of the cervix for

mifepristone) have been combined with an misoprostol) to produce Arboprost tromethamine –analog of PGF2"

PGE2 and PGF2#  should be superior to oxytocin for inducing

NSAIDs successfully inhibit the formation of these prostaglandins Aspirin is also effective in dysmenorrhea, but because it has low

Alprostadil

Penile pain is a frequent side effect, which may be

Pulmonary HPN

PGI2  lowers peripheral, pulmonary, and coronary epoprostenol)

Iloprost-prostacyclin analogs used also for pHPN

Treprostinil-Peripheral Vascular Disease

PDA

PGE2  is a powerful bronchodilator when given in aerosol COUGHING

PGF2! and TXA2 are both strong bronchoconstrictors and

ASTHMA. zafirlukast, montelukast)

(6)

GASTROINTESTINAL SYSTEM

!

- approved indication is for prevention of NSAID-induced peptic ulcers

IMMUNE SYSTEM

! !

rejection process

! TXA2 increases during acute rejection !

of acute rejection because of their lymphotoxic effects, inhibit both phospholipase and COX-2 activity.

! Inflammation

!  Aspirin has been used to treat arthritis ! Rheumatoid Arthritis

! Eicosanoids- amplify the inflammatory response

GLAUCOMA

!

first prostanoid used for glaucoma

o Other drugs are: and

o  Adverse effects include irreversible brown

pigmentation of the iris and eyelashes, drying of the eyes, and conjunctivitis

Misoprostol-orally active synthetic analog of PGE1

Cell- Mediated Organ Transplant Rejection

PGI2  to renal transplant patients has reversed the

Corticosteroids- the first- line drugs used for treatment

Latanoprost-a stable long-acting PGF2" derivative, was the

bimatoprost, travoprost, unoprostone

References

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