EICOSANOIDS

Biochemistry Symposium

2023
OVERVIEW

• INTRODUCTION
• BIOSYNTHESIS
• REGULATION & INHIBITION
• CELLULAR RECEPTORS AND SIGNALING PATHWAY OF
ECOSANOIDS
• PHYSIOLOGIC FUNCTIONS OF EICOSANOIDS
• CLINICAL IMPLICATIONS AND THERAPEUTIC
POTENTIAL OF EICOSANOIDS
EICOSANOIDS
• They are 20 C compounds (Greek, eicosa = twenty).
• It is derived from arachidonic acid.
• The term is now used to describe a family of closely
related derivatives of hypothetical C20 molecule named
Prostanoic acid
• They are extremely potent compounds that elicit a wide
range of responses, both physiologic (inflammatory
response) and pathologic (hyper - sensitivity).

Pr ostanoic acid
EICOSANOIDS
Although they bear actions similar with hormones, they
differ from true hormones:
• Produced in small amounts in almost all tissues
rather than in specialized glands
• Acts locally rather than being transferred through the
blood
• Very short lived; rapidly catabolized; and are not
stored
 PROSTAGLANDINS
• 1st discovered in human semen by Ulf Von Euler in
1930; were found to stimulate uterine contraction
and reduce blood pressure
• Presumed to be synthesized by prostate gland hence the
0 name
• Later realized were synthesized in all tissues except
erythrocytes.
• It has a cyclopentane ring ( formed by carbon atoms 8
to 12) and two side chains, with carboxyl group on one
side.
• Prostaglandins differ in their structure due to substituent
group and double bond on cyclopentane
PROSTAGLANDINS - NOMENCLATURE

• Abbreviated as PG, with the class designated by a capital

letter A,B,D,E,F,C,H and I, followed by a number.
• PGE and PGF; 1st isolated from the biological fluids
• The letters refer to the different ring structure, except in PGG
and PGH: same ring structure (cycloendohydroperoxide).
• In the same series, depending upon double bonds on the
side chains designated as PGE1, PGE2, PGE3..etc
THROMBOXANES (TXAs):

• Named so because they are identified first in thrombocytes.

• Structure is similar to PGs, but have an oxygen atom in
the cyclic ring and contains a six numbered heterocyclic
oxane ring.
• The most common thromboxane, TXA2, contains an
additional oxygen atom attached both to carbon 9 and
carbon 11 of the ring
• TXB2- a stable degradation product of TxA2
Leukotrienes:
• are the newly discovered family of conjugated trienes formed
from arachidonic acid in leucocytes, mast cells, and
macrophages by the lipoxygenase pathway, in response to
both immunologic and non-inflammatory stimuli.
• Each LT product has a distinct role in promoting different aspects
of inflammation.
• LTB4 attracts and activates neutrophils, monocytes, and
lymphocytes, contributing to the increase in activated leukocytes
that is a hallmark of tissue inflammation
• The cysteinyl LTs (LTC4, LTD4, and LTE4) cause plasma leakage
from postcapillary venules, leading to the edema that is also
characteristic of inflammation
BIOSYNTHESIS OF EICOSANOIDS:

 The lipid mediators, prostaglandins and leukotrienes, are

produced from arachidonic acid (AA) present in membrane
phospholipids
 Mechanical, chemical, and physical stimuli or other
mediators release AA from membrane phospholipids through
the action of cellular phospholipases, mainly phospholipase
A2
 The biochemical signals involved in the activation of
phospholipase A2 include an increase in cytoplasmic Ca2+
and activation of various kinases in response to external
stimuli.
 CYCLOOXYGENASE PATHWAY- CYCLIC
PATHWAY (SYNTHESIS OF PROSTAGLANDINS
AND THROMBOXANES)
• In humans, the most important precursor for prostaglandins is
arachidonic acid, a polyunsaturated fatty acid with four
double bonds (eicosatetraenoic acid).
• It isstored in cell membranes as the C2 ester of
phospholipids

Site: In all types of mammalian cells except RBCs

(no cyclooxygenase activity has been found in human RBCs

SYNTHESIS OF P G H 2 :
• 1st step in prostgladin synthesis is oxidative cyclization
of free arachidonic acid to yield PGH2, by prostagladin
endoperoxide synthase (PGH Synthase).

PGH synthase exhibits 2 catalytic activities

Cyclooxygenase (COX) and Peroxidase

Initial step catalyzed by cyclooxygenase and forms the

five- membered ring with the addition of 4- atoms of
oxygen. 2 between C- 9 and 11, and 2 at C-15 to form
an unstable endoperoxide,PGG2.

The hydroperoxy group at carbon 15 of PGG 2 is quickly

reduced to a hydroxyl group by a peroxidase to form
another endoperoxide, PGH2.
ISOZYMES OF PGH SYNTHASE:

Cyclooxygenase (COX): two isozymes usually denoted as COX-1 and COX-2

CONVERSION OF PRIMARY PROSTAGLANDIN
( P G H ) TO OTHER EICOSANOIDS

• Cell specific enzymes like PGE synthase and PGD

synthase and so on are involved
• Thus, vascular endothelium produces PGE and PGI,
and platelets produce thromboxanes (TXAs).
• In kidney and spleen, isomerase catalyses production
of PGE2 and reductase catalyses production of PGF2
 REGULATION OF PROSTAGLANDIN SYNTHESIS

• The synthesis of prostaglandins can be inhibited by various compounds.

• Cortisol, a steroidal anti-inflammatory agent, inhibits phospholipase A2 activity,
preventing the release of arachidonic acid from membrane phospholipids, which is a
precursor for prostaglandin synthesis
• Aspirin, indomethacin, and phenylbutazone, nonsteroidal anti-inflammatory agents
(NSAIDs), inhibit both COX-1 and COX-2 enzymes, blocking the synthesis of the
parent prostaglandin, PGH2.
• Systemic inhibition of COX-1 by aspirin can lead to toxicity, including stomach and
kidney damage and impaired blood clotting.
• COX-2-specific inhibitors like celecoxib were developed to reduce pathological
inflammation while preserving the physiological functions of COX-1. However,
their use has been associated with an increased risk of heart attacks.
SYNTHESIS OF LEUKOTRIENES:

• The AA extracted from phospholipids is acted on by the

first committed enzyme in the LT synthetic pathway, 5-
lipoxygenase (5-LO)
• 3 different lipoxygenases (dioxygenases) insert oxygen
into the 5, 12, and 15 positions of arachidonic acid,
giving rise to hydroperoxides (HPETE).
• Depending on the position of addition, 3 types of HPETE
have been found: 5-HPETE, 12-HPETE, and 15-HPETE
• Only 5-HPETE is converted to a series of leukotrienes, the
nature of which varies according to the synthesizing
tissue.
Cellular Receptors and signaling Pathways

• The reception of eicosanoids is generally associated with G

protein-coupled receptors (GPCRs)

• Targeting of GPCRs that are involved with eicosanoids is

hence widely utilized for therapeutic intervention; GPCRs
correspond to 30% of all identified drug targets and remain
major targets for new drug development.
 Signaling by GPCRs
 They are associated with heterotrimeric G proteins (Gαβγ)
 Ligand binding induces conformational changes, activating G proteins
 When it is inactive, the α subunit of the G protein is bound to
guanosine diphosphate (GDP)
 when a GPCR is activated, it induces the α subunit to release GDP and
instead binds to guanosine triphosphate (GTP)
 The exchange of GDP with GTP results in a conformational change in
the G protein, which leads to its activation
 The α subunit of G protein has a GTPase activity, and once it
hydrolyzes GTP to GDP, it once again becomes inactive
There are three important pathways by which GPRS are involved in
signal transduction associated with eicosanoids: Gs, Gi, and Gq .
Examples
Gs:
 IP for PGI2
 EP2 for PGE3
Gi:
 EP3 for PGE2

Gq:
 G11, G15, and G16 for TAX2
 EP1 for PGE2,
 and FP for PGF2 alpha
 Physiological Functions of Eicosanoids
in Inflammation
 Eicosanoids are local hormones with short half-lives, acting as autocrine and
paracrine mediators.
 Found in most living things, they play a crucial role in inflammation, immunity,
and central nervous system signaling.
 Eicosanoid receptors are predominantly G protein-coupled receptors.
 Enzymatic conversion by cyclooxygenases (COX) and lipoxygenases (LOX) in
response to inflammatory stimuli.
 Triggered by various factors such as tissue injury, infection, or immune cell
activation
 Initiation of Inflammation

 Eicosanoids serve as signaling molecules in the

inflammatory response.
 Produced by immune cells (macrophages,
neutrophils) and non-immune cells (endothelial
cells, fibroblasts).
 Tight regulation to ensure a specific and appropriate
response to inflammatory stimuli.
 Prostaglandins in Inflammation

 Prostaglandins, a major class of eicosanoids, are produced by COX

enzymes.
 Examples include prostaglandin E2 (PGE2) and prostaglandin D2
(PGD2).
 Bind to specific receptors on target cells, triggering intracellular
signaling pathways that promote inflammation.
 Prostaglandins induce vasodilation and increased vascular
permeability.
 Result in enhanced blood flow to the site of inflammation and
facilitate immune cell migration into the tissue.
 Stimulate synthesis and release of other inflammatory mediators,
amplifying the immune response
Leukotrienes in Inflammation

 Eicosanoids derived from the LOX pathway, such as

leukotrienes, contribute to inflammation.
 Leukotrienes activate and recruit immune cells,
particularly leukocytes, to the inflamed tissue.
 Potent chemoattractants for neutrophils and other
immune cells
 Vasodilation and Increased
Vascular Permeability
 Eicosanoids, especially prostaglandins, induce vasodilation and
increased vascular permeability during inflammation.
 These effects are crucial for initiating and progressing the immune
response.
 Prostaglandins, like PGE2, promote vasodilation by acting on
smooth muscle cells surrounding blood vessels.
 Specific receptors activation leads to smooth muscle relaxation and
subsequent vessel dilation.
 Increased blood flow brings oxygen, nutrients, and immune cells to
the inflamed area
 con’t
 Prostaglandins act on endothelial cells, triggering signaling pathways that
rearrange cell-cell junctions.
 Loosening of tight junctions creates openings in vessel walls, allowing
fluid, proteins, and immune cells to enter the tissue.
 Fluid leakage contributes to swelling (edema) observed during
inflammation
 Vasodilation and increased permeability ensure adequate blood supply,
aiding in immune cell recruitment.
 Oxygen, nutrients, and immune mediators are delivered to support
immune response and tissue repair
 Recruitment of Immune Cells

 Eicosanoids, including prostaglandins and leukotrienes, play a critical role

in immune cell recruitment to inflammation sites
 Eicosanoids act as chemoattractants, guiding immune cells towards areas
of injury or infection.
 Released during inflammation, eicosanoids establish concentration
gradients, directing immune cell migration
 Immune cells possess specific receptors for eicosanoids (e.g., leukotriene
receptors on leukocytes, prostaglandin receptors on macrophages).
 Binding triggers intracellular signaling, activating cells and promoting
migration.
 con’t

 Eicosanoids enhance adhesion of immune cells to endothelial

cells, facilitating subsequent transmigration.
 Influence endothelial cell junction integrity, allowing immune
cells to pass through vessel walls
 Recruited immune cells (neutrophils, macrophages) play key
roles in pathogen clearance, debris removal, and tissue repair.
 Eicosanoids contribute to a coordinated immune response
required for inflammation resolution and tissue homeostasis
 Regulation of Inflammatory
Mediators
 Eicosanoid Influence:
 Eicosanoids regulate the production and release of inflammatory
mediators (cytokines, chemokines, adhesion molecules).
 Acts in autocrine or paracrine manner, shaping local inflammatory
microenvironment.
 Inflammation Regulation:
 Essential for preventing excessive tissue damage during immune
response.
 Specialized pro-resolving mediators (SPMs), derived from
eicosanoids, crucial in resolution phase
 con’t
 SPMs in Resolution:
 During resolution, pro-inflammatory eicosanoid production decreases,
SPM synthesis increases.
 SPMs (lipoxins, resolvins, protectins, maresins) have potent anti-
inflammatory and pro-resolving properties.
 SPM Actions:
 Promote clearance of inflammatory cells, especially neutrophils.
 Facilitate tissue repair, restore tissue integrity.
 Regulate cytokines, chemokines, and other mediators to prevent excessive
inflammation.
 Blood Clotting and Platelet
Aggregation
 Role of Eicosanoids:
 Eicosanoids, especially thromboxanes, crucial in platelet aggregation and blood
clotting.
 Platelet Activation:
 Platelets, when activated by vessel injury, synthesize thromboxane A2 (TXA2) from
arachidonic acid.
 TXA2 acts on thromboxane receptors, promoting platelet activation and aggregation.
 con’t
 Platelet Functions:
 TXA2 induces shape change, leading to adhesion and formation of platelet plug.
 Stimulates granule release, including ADP, amplifying platelet activation and
aggregation
 Formation of Stable Clot:
 Eicosanoids contribute to the formation of a stable blood clot, preventing
excessive bleeding.
 Platelet aggregation forms a meshwork that seals damaged vessels
 con’t
 Regulation and Balance:
 Balance between thromboxane A2 and antiplatelet substances like
prostacyclin (PGI2) is crucial.
 Disruptions may lead to thrombotic disorders.
 Clinical Implications:
 Antiplatelet drugs, like aspirin, inhibit COX-1, reducing thromboxane A2
production.
 Understanding eicosanoid role vital for managing clotting disorders and
ensuring proper hemostasis
 Resolution of Inflammation

 Role in resolving inflammation as inflammatory response

subsides.
 Specialized lipid mediators (lipoxins, resolvins, protectins)
actively promote resolution.
 Counteract pro-inflammatory signals and facilitate tissue repair
processessLipid mediators derived from eicosanoids contribute to
tissue homeostasis.
 Prevent chronic inflammation and support the return to normal
physiological function.
 Prostaglandins in Platelet
Homeostasis
 Thromboxane A2 (TXA2) Production:
 Produced by COX-1 in activated platelets.
 Promotes adherence, platelet aggregation, and vascular smooth
muscle contraction, leading to thrombus formation.
 Prostacyclin (PGI2) Production:
 Produced by COX-2 in vascular endothelial cells.
 Inhibits platelet aggregation, stimulates vasodilation, impedes
thrombogenesis.
 con’t
 Balancing Thrombi Formation:
 TXA2 and PGI2 have opposing effects, limiting thrombi formation to sites
of vascular injury.
 Aspirin's antithrombogenic effect: irreversible acetylation of COX-1 in
platelets and COX-2 in endothelial cells.
 Mechanism of Low-Dose Aspirin Therapy:
 COX-1 inhibition in platelets cannot be overcome, preventing TXA2
synthesis.
 COX-2 inhibition in endothelial cells can be overcome due to nuclear
presence, maintaining PGI2 synthesis.
 Basis for low-dose aspirin therapy to lower stroke and heart attack risk by
decreasing thrombi formation
 Clinical Implications and Therapeutic
Potential
 Bioactive Lipid Mediators:
 Crucial in inflammation, immune response, and various physiological
processes.
 Significant clinical implications and therapeutic potential.
 Role in Inflammatory Diseases:
 Prostaglandins, leukotrienes, and thromboxanes from arachidonic acid
mediate inflammation.
 Key in conditions like rheumatoid arthritis, asthma, and inflammatory
bowel disease
 con’t

 NSAIDs and Eicosanoid Synthesis:

 NSAIDs (aspirin, ibuprofen) inhibit cyclooxygenase, blocking prostaglandin
synthesis.
 Effective in treating pain and fever.
 Cyclooxygenase Forms:
 COX-1 present constitutively in most tissues, serves homeostatic functions.
 COX-2 induced by inflammatory stimuli; targeted by COX-2 inhibitors.
 COX-2 inhibitors aim to block harmful inflammation while preserving COX-1's
protective effects.
 con’t

TxA2 and PGI2 in Cardiovascular System:

 TxA2 (Thromboxane A2):
 Released from activated platelets, promotes platelet aggregation and
vasoconstriction.
 Implicated in thrombotic and ischemic diseases.
PGI2 (Prostacyclin):
 Produced by COX-2 in vascular endothelial cells.
 Inhibits platelet aggregation, promotes vasodilation.
Balancing Act:
 Maintaining balance crucial for preventing excessive clotting and maintaining blood
flow.
 Cardiovascular implications highlight the delicate interplay between TxA2 and PGI2.
 Eicosanoids in Asthma

Leukotrienes and Prostaglandins:

 Central in asthmatic reactions, acting as bronchoconstrictors, inhibiting
mucociliary clearance, increasing blood flow, and inducing inflammation.
Management Strategies:
 Targeting leukotrienes and prostaglandins crucial in asthma.
 A key strategy involves modulating eicosanoids and their pathways.
 Pharmacological Modulation of
Eicosanoid Pathways
Aspirin and NSAIDs:
 Inhibit both COX-1 and COX-2.
 Irreversibly acetylate and inactivate cyclooxygenases.
 Efficacious in treating pain and fever.

Selective COX-2 Inhibitors:

 200-300 fold more potent in blocking COX-2 than COX-1.
 Theoretical benefits in reducing gastric ulceration.
 Risks include potential cardiovascular and cerebrovascular events due to altered
prostacyclin (PGI2) and thromboxane A2 (TxA2) balance.
con’t

5-Lipoxygenase Pathway:
 Not affected by NSAIDs.
 New inhibitors developed; e.g., Zileuton.
 Useful in treating asthma by inhibiting leukotriene
production.
con’t

Corticosteroids in Inflammation Management

 Reduce transcription of genes encoding COX-2, phospholipase A2,
pro-inflammatory cytokines (IL-1, TNF), and iNOS.
 Widely used for their potent anti-inflammatory effects.

Blocking Leukotriene Receptors:

 Block leukotriene receptors, preventing their actions. E.g.,
Montelukast.
 Useful in treating asthma by inhibiting leukotriene actions
 Summary
 Eicosanoids are a diverse group of bioactive lipid molecules that play crucial roles in
various physiological and pathological processes within the human body.
 Eicosanoids are classified into three major groups based on their chemical structure
and biological functions: prostaglandins, thromboxanes, and leukotrienes.
 Prostaglandins are involved in inflammation, blood clotting, and regulation of blood
pressure
 Thromboxanes contribute to platelet aggregation and blood clot formation.
 Leukotrienes are associated with allergic and inflammatory responses, particularly in
the respiratory system.
 Cont’d
 These lipid mediators are synthesized and released by various cell types,
including immune cells, platelets, and endothelial cells, in response to specific
stimuli such as injury, infection, or inflammation.
 Although eicosanoids are essential for normal physiological functions, their
dysregulation has been implicated in the pathogenesis of numerous diseases,
including asthma, cardiovascular diseases, and inflammatory disorders.
 pharmacological interventions targeting eicosanoid synthesis or receptor
signaling pathways have been developed to treat these conditions.
 In summary, eicosanoids are bioactive lipid molecules derived from
polyunsaturated fatty acids, with diverse roles in inflammation, blood clotting,
immune responses, and other physiological processes.