Second

Messenger
Systems
Dr. Kunal A. Chitnis
2nd Yr Resident
T.N.M.C.
16th July 11

 Molecules that relay signals from receptors on the cell surface

to target molecules inside the cell i.e. cytoplasm or nucleus
Relay the signals of hormones like epinephrine, growth factors
& others; causing some kind of change in the activity of the
cell
The term was coined upon the discovery of these substances
in order to distinguish them from hormones & other molecules
that function outside the cell as first messengers in the
transmission of biological information

 Earl Wilbur Sutherland Jr.

discovered second messengers
won the 1971 Nobel Prize in Medicine
He saw that epinephrine would
stimulate the liver to convert glycogen
to glucose in liver cells, but
epinephrine alone would not convert
glycogen to glucose
He found that epinephrine had to trigger a
second messenger, cyclic AMP for the liver to convert
glycogen to glucose

cAMP
System

Ligands

Epinephrine ANP, NO
Ach

Primary
Effector

Adenyl
cyclase

Secondary cAMP
messenger

cGMP
System

Phosphoinositol
System

Tyrosine
Kinase
System

Oxytocin

PDGF

Guanylate Phosphocyclase
lipase C

Receptor
Tyrosine
kinase

cGMP

IP3 & DAG;

Ca2+

G- Proteins
Guanine nucleotide binding proteins which act as a Transducer
between a receptor & an effector
Discovered by Alfred Gilman & Martin Rodbell in 1990
Significance:
Of the top 100 drugs,
26 directed at GPCRs
60 % act through GPCRs
40 % of prescriptions
3rd largest family of genes (865)
Present in almost every organ system

Endogenous Ligands:
Sensory signal mediators: Light & Olfactory stimulatory molecules
Biogenic amines: Dopamine, Adr, NA, Ach, Histamine
Peptide hormones: FSH, GnRH, calcitonin, TRH, Oxytocin
Mediators of inflammation: Prostaglandins, PAF, leukotrienes,
chemokines

Structure:
Embedded in the plasma membrane

Lysozyme
insert

7 transmembrane -helices
Rhodopsin was first of these
to have its structure confirmed by
X-ray crystallography

ligand

-Adrenergic
Receptor

PDB 2RH1

Binding Domains
Small molecular ligands
bind to sites within the
hydrophobic core formed by
transmembrane helices
Protein and peptide agonists
bind to N terminus &
extracellular hydrophilic
loops joining the
transmembrane domain
G-proteins bind either to
second and third
cytoplasmic loop which is
largest or to carboxy terminus

 Molecular Switch: On/Off

Heterotrimeric
-subunit
-subunit
-subunit
subunit: specific recognition of
receptors & effectors;
GTP binding site
subunit: Membrane localisation by prenylation of subunit
Coded by of genes:
23, -7, -12

Functions of subunit & dimer

Receptors

Couplers

Muscarinic

Gi, Go, Gq

Dopamine D2

Gi, Go

-adrenergic

Gs, Gi

2-adrenergic

Gi, Gs, Go

GABA-B

Gi, Go

5-HT

Gi, Gq, Gs

G Protein Activation
Conformational change
in receptor
Transmitted from ligand
binding pocket to 2nd & 3rd
intracellular loop
subunit exchange GDP
with GTP
Presence of GEFs
(Guanine exchange factors)
Release of GTP bound
subunit & dimer

Inactivation
Activated subunit is
inactivated by hydrolysis
of GTP to GDP by GAPs
(GTPase Activating
Proteins)
Rebinds to complex
Modulated by
Regulators of G
proteins Signaling (RGSs)
Acclerate hydrolysis of
GTP & potential drug
targets

Toxins
Cholera toxin catalyzes covalent modification of Gs
ADP-ribose is transferred from NAD+ to an arginine residue
at the GTPase active site of Gs
ADP-ribosylation prevents GTP hydrolysis by Gs
The stimulatory G-protein is permanently activated.
Pertussis toxin catalyzes ADP-ribosylation at a cysteine residue
of the inhibitory Gi, making it incapable of exchanging GDP for
GTP
The inhibitory pathway is blocked.
ADP-ribosylation is a general mechanism by which activity of
many proteins is regulated, in eukaryotes & prokaryotes.

Receptor Desensitization
Activated receptor is
phosphorylated via a
G-protein Receptor Kinase
Binds to a protein arrestin
promotes removal of the
receptor from the membrane by
clathrin-mediated endocytosis
May also bind a cytosolic
Phosphodiesterase, bringing
this enzyme close to where
cAMP is being produced,
contributing to signal turnoff

Adenyl Cyclase- cAMP Pathway

Adenyl Cyclase
9 membrane bound, 1 soluble isoform (AC1-10)
120kDa
Basal enzymatic activity modulated by
GTP liganded subunit Gs & Gi
Other regulatory interactions: subunits, Ca2+, protein
kinase & diterpene forskolin
Dephophorylation of ATP by removal of 2 phosphate
molecules cAMP

cAMP has following major targets:

1. cAMP dependent Protein Kinase A (PKA)
2. cAMP regulated Guanine nucleotide exchange factors
termed EPACs (Exchange Proteins Activated by cAMP)
3. CREB (cAMP responsive element binding protein)

Protein Phosphorylation most common form of posttranslational modification in nature

Protein function altered by addition of a negatively charged
phosphate group to a Ser, Thr or Tyr residue by Protein Kinase:
Binding properties
Enzymatic activity if a catalytic protein
Protein phosphatase catalyzes removal of the Pi by hydrolysis
O

Protein Kinase
OH + ATP

Protein

Protein

P
O

Pi

H2O

Protein Phosphatase

O + ADP

1. Protein Kinase A
. Holoenzyme: 2 Regulatory(R) & 2 Catalytic(C) subunits
. Heterotetramer complex R2C2
. cAMP, R inhibits C inactive
. cAMP 4 cAMP mols bind to R2C2, 2 to each R
lowers
affinity to C Activation

Active C subunits
Phosphorylate serine/threonine residues on proteins
As protein expression varies from cell type to cell type,
proteins that are available for phosphorylation will depend upon
the cell type
Phosphorylation of ion channels Regulation
(Ca2+ activated K+ channel activation & Na+/K+ ATPase)
Inhibition of Myosin Light Chain Kinase

Actions of Protein kinase A

Cell Type

Stimulators

Inhibitors

Effects

Hepatocyte

Epinephrine(),
Glucagon

Produce glucose:
stimulate glycogenolysis,
inhibit glycogenesis,
stimulate gluconeogenesis,
inhibit glycolysis.

Skeletal
-Myocyte

Epinephrine()

Produce glucose:
stimulate glycogenolysis,
inhibit glycogenesis,
stimulate glycolysis

Cardiomyocyte

Norepinephrine
()

Sequester Ca2+ in
sarcoplasmic reticulum,
Phosphorylates
phospholamban

Smooth muscle
myocyte

2 agonist (2)
Histamine (H2)
Prostacyclin
Prostaglandin
D2/E2

Adipocyte

Epinephrine (),
Glucagon

Enhance
lipolysis

Neurons in
Nucleus
accumens

Dopamine (D3)

Activate
reward system

Principal Cells

Vasopressin
(V2)

Synthesis &
Exocytosis of
Aquaporin 2

Juxtaglomerular Adrenergic (1)

Cells
Dopamine(D1)
Glucagon

Muscarinic Vasodilation
(M2)

Renin
secretion

2. cAMP Response
Element-Binding
(CREB)
. Cellular transcription factor
. Genes: c-fos, the neurotrophin
BDNF, tyrosine hydroxylase &
many neuropeptides
. Neuronal plasticity & long-term
memory formation
. Development & progression of
Huntingtons Chorea

3. Exchange Proteins Activated

by cAMP (EPAC)
. cAMP Regulated Guanine Nucleotide
Exchange Factors
. Bind to GDP liganded GTPase,
exchange of GDP for GTP
. Activation of PKC
. Cell differentiation/proliferation, cytoskeletal
organization, vesicular trafficking & nuclear transport
. Additional effector system
. Potential target for cancer therapy

Therapeutic Applications
Selected Drugs that target cAMP signalling
Drug

MOA

cAMP

Therapeutic
Application

Salmeterol

2 Agonist

Asthma, COPD

Rimonabant

CB-1 Antagonist

Obesity

Haloperidol

D2,D3,D4 Antag

Schizophrenia

Metoclopramide D2, 5HT4 Antag

Nausea,
Vomiting

Desmopressin

Diabetes
insipidus

V2 Receptor
Agonist

Metoprolol

1 Antag

Angina,
Hypertension,
CHF

Morphine

Agonist

Pain

Sumatriptan

5-HT1D/5-HT1B

Migraine

Ibuprofen

Non selective
Inhibitor of COX

Inflammation,
Pain

Ranitidine

H2 Antag

Peptic ulcer,
GERD

Misoprostol

PG Receptor
Agonist

Prevention of
NSAID ulcers

Cabergoline

D2 Agonist

Parkinsons
disease

Novel Drug Targets:

Analgesia

cAMP nociception
AC1 & AC5 involved
Selective AC1 & AC5inhibitors Analgesics
Preclinical stages

Drug dependence
Repeated opiod exposure upregulation of AC activity
3 specific isoforms- AC1, AC5 & AC8
Selective inhibitors for opiod dependence

Neurodegenerative disorders
Target second messengers used my multiple neurotransmitters
AC1 is most attractive target
AC1 activators used for cognitive decline

 Chronic Heart Failure:

Alterations in adrenoreceptor- AC- cAMP pathway
Downregulation of 1 receptors
Upregulation of inhibitory G proteins &
G-Protein coupled receptor kinases
Catecholamine refractoriness, Exercise intolerance
Protective phenomenon shields myocytes from
arrhythmogenic, hypertrophic & apoptotic effects of
catecholamines
Genetic variant of G-Protein Coupled Receptor Kinase 5
(GRK5)Accelerated desensitization Better prognosis

Sperm function:
sAC regulates sperm motility & capacitation
cAMP mediates capacitation
Inhibitors for male contraception in early stages

Cyclic GMP Pathway

 Unlike cAMP, cGMP has established signaling roles in only

few cell types
Signaling pathways that regulate synthesis include:
1. Nitric Oxide
2. Hormonal Regulation (ANP/BNP/CNP)

1. Nitric Oxide
.
.
.
.

Nitric oxide: diatomic free radical gas

Lipid soluble
Very small easy passage between cell membranes
Short lived, degraded or reacted within a few seconds

. Synthesis:
Synthesized from L-arginine catalyzed by
reaction of NO-synthase
to form NO and L-citrulline

 Nitric Oxide Synthase (NOS) exists as 4 isoforms

neuronal type I isoform (nNOS)
inducible type II isoform (iNOS)
endothelial type III isoform (eNOS)
mitochondrial isoform (mtNOS)

NOS constitutively expressed in:

eNOS :
Endothelium, cardiac myocytes, renal mesangial cells,
osteoblasts, platelets
nNOS :
CNS and NANC nerves

Control exerted in following ways:

Endothelium-dependent agonists (Ach, bradykinin, substance P)
cytoplasmic concentration of Ca2+ calcium-calmodulin
eNOS or nNOS activation
Shear stress in resistance vessels Sensed by
mechanoreceptors Transduced via a serine threonine protein kinase
called Protein kinase B/ Akt Phosphorylation of specific residues on
eNOS

 iNOS:
Macrophages, Kuffer cells, neutrophils, fibroblasts, vascular
smooth muscle cells & endothelium
Activity independent of Ca2+
Positive inducers : IFN, TNF, IL-1, IL-2, LPS, antigens
Inhibitory cytokines: TGF-, IL-4, IL-10, glucocorticoids

2. Natriuretic Peptide Receptors

3 small peptide ligands:
. Atrial Natriuretic Peptide (ANP):
. Released from atrial storage granules
. Intravascular volume/ Stimulation with pressor
hormones
. Brain Natriuretic Peptide (BNP):
. Synthesized and released from ventricular tissue
. Volume overload

 C-type Natriuretic Peptide (CNP):

Synthesized in brain & endothelial cells
Growth factors/ stress on vascular endothelial cells
Major physiological effects:
BP (ANP/BNP)
Cardiac hypertrophy & fibrosis (BNP)

Receptors with intrinsic enzymatic activity

3 types of receptors:
1) ANP Receptor (NPR-A)
. Binds to ANP & BNP
. Maintaining normal state of CVS
2) BNP Receptor (NPR-B)
. Binds to BNP
. Role in bone function
3) CNP Receptor (NPR-C)
. No enzymatic activity

 Ligand brings juxtamembrane regions together

Phosphorylation of serine residues
Stimulation of guanyl cyclase

Guanylate Cyclase:
Membrane-bound (type 1) & soluble (type 2) forms
Membrane bound activated by ligands
Soluble activated by NO
Catalyzes reaction of
GTP to cGMP

Downstream reactions of cGMP:

Activation of Protein Kinase G
cGMP gated ion channels
cGMP-modulated Phosphodiesterase

cGMP-dependent protein kinase or Protein Kinase G

(PKG)
Serine/threonine kinases
PKG I cytoplasm, PKG II membrane associated
PKG I smooth muscles, platelets, brain
PKG II intestines, bones, kidney
Regulatory and catalytic domains contained on a single
polypeptide chain
Upon activation dimerizes to form PKG holoenzyme

 Actions of PKG I:
Inhibits of Gq/G11 Inhibition of Phospholipase C Ca2+
Inhibits G12/G13 Inhibits Ca2+ sensitizing mechanisms
Phosphorylates and inhibits the Myosin light chain

kinase which normally phosphorylates the myosin light

chains

 Effects:
Relaxes all smooth muscle types
Inhibits platelet aggregation & granule secretion
Functions of PKG II:
Stimulates in chloride & water secretion in small intestine
Normal endochondral bone development
Inhibits renin secretion

Therapeutic Applications
1. NO Donors
. Organic nitrates
.

Short acting: Nitroglycerin (NTG)

Long acting: Isosorbide mononitrate, Isosorbide dinitrate,

Pentaerythritol tetranitrate
.

Use: Treatment of Angina

A/E: Tolerance (Nitrate free interval)

Sodium Nitroprusside

Hypertensive emergencies

Nicorandil
K+ channel opner and NO donor
Antianginal

CCBs: Nitrendipine, Nifendipine, Lacidipine

Dihydropyridine
Release NO
Retard atherosclerosis
Use : Hypertension, angina

 Blockers: Nebivolol, Celiprolol

Additional vasodilatory effects

2. Natriuretic Peptides
.
.
.
.

Nesiritide:
Synthetic Brain Natriuretic Peptide
Promotes vasodilation, natriuresis & diuresis
Use: Acutely decompensated congestive heart failure

. Ecadotril:
.
.
.

Neural Endopeptidase (NEP) catalyses BNP degradation

It inhibits NEP
Use: CHF (Phase 2)

BAY series of compounds

8-bromo-cGMP
8-bromoPET-cGMP
8-pCPT-cGMP
Specific activators & inhibitors of cGKI & cGKII
Improve the specificity & availability of treatment
Evaluated for:
Asthma
Graft survival after liver & lung transplantation

Phosphodiesterases (PDE)
PDE comprise family of enzymes expressed in almost all cells
of the body & of prime importance in cellular functioning
Hydrolysis of phosphodiester bond in the second messengers
c AMP & c GMP inactive forms 5 AMP and 5 GMP
respectively

 Reversal of activation of cellular protein kinases

Inactivation leads to termination of intracellular signals
Regulators of cyclic nucleotide signaling & responsible for
diverse physiological functions
PDEs comprise a super family composed of 25 genes
Categorized into 11 sub families i.e. PDE 1 PDE 11
Substrate specificity:
c AMP
Specific
PDE 4
PDE 7
PDE 8

c GMP
Specific
PDE 5
PDE 6
PDE 9

cAMP & cGMP

Specific
PDE 1
PDE 2
PDE 3
PDE 10
PDE 11

 Act by modulating the levels of 2nd messengers

Therapeutic Applications:
Congestive heart failure:
PDE-3 Inhibitors Amrinone, Milrinone
In force of contraction
Direct vasodilation of both the resistance & capacitance vessels
Inodilators

 Erectile dysfunction:
PDE-5 Inhibitors Sildenafil, Tadalafil, Vardenafil
Never combined with nitrates:
Nitrates produce vasodilatation by NO dependent elevation
of cGMP in vascular smooth muscle
Thus PDE 5 inhibitor if given along with of a NO donors can
cause profound & extreme hypotension
Pts should be asked for a history of nitrate consumption
within
previous 24 hrs

 Bronchial Asthma:
Theophylline, Aminophylline
Non selective PDE inhibitor
Narrow therapeutic range (5 20 mcg/ml)
Other uses: COPD, Premature apnoea in infants

Peripheral Vascular Disease:

Pentoxyphylline
PDE 3 inhibitor
Rheologic modifier improves the flexibility of RBCs
& blood viscosity
Improves microcirculation
Cilastazol
PDE 3 inhibitor
Promotes vasodilatation & inhibition of platelet aggregation

 Antiplatelets
Dipyridamole
Inhibits PDE 5
Prevents uptake & degranulation of adenosine
Was introduced for angina pectoris but was a failure due to
coronary steal phenomenon
Anagrelide
PDE 3 inhibitor
Other uses: Essential thrombocytosis,
Thrombocytopenia in Polycythmia Vera
Zaprinast
PDE 6 Inhibitor
Antiproliferative & proapoptic property
Vasoproliferative disorders

 Antispasmodics
Drotaverine
Inhibits PDE 4
Selective for smooth muscles
Intestinal, biliary, renal colic; Irritable bowel syndrome;
Dysmenorrhea; Acceleration of labor

Phospholipase C: IP3-DAG Pathway

Three Types of Inositol phospholipids:

PI, PI(4)P, PI(4,5)P2

Phospholipase C exists as 2 isoforms:

1. Phospholipase C:
.
Activated by GPCRs couple to Gi/Gq release GTP
bound
subunit & dimer both activate
2. Phospholipase C:
.
Activated by Receptor/Non Receptor Tyrosine Kinases
. Cytosolic enzymes
. Translocate to plasma membrane on receptor activation
. Ligands:
AGT, GnRH, GHRH, Oxytocin, TRH

Phospholipase C forms Diacylglycerol & Inositol 1,3,5- triphosphate

from Phosphotidyl Inositol 4,5-bisphosphate

Protein kinase C:
Regulatory domain & catalytic
domain tethered together by a
hinge region
C1 domain, present in all of the
isoforms of PKC has a binding site
for DAG
C2 domain acts as a Ca2+ sensor
Catalytic Region brings about
phosphorylation Ser/Thr a.a. of
proteins
Upon activation, translocated to the

Cell type

Activators

Effects

Smooth muscle
(vascular)

5HT(5HT2A)
Adrenergic(1)

Vasoconstriction

Smooth muscle
(GIT)

5HT(5HT2A/5HT2B)
Adrenergic(1)

Contraction
Bronchoconstriction

Smooth muscle
(bronchi)

5HT(5HT2A)
Adrenergic(1)
Ach(M1/M3)

Smooth muscle
(ureter/ urinary bladder/
urethral sphincter)
Smooth muscle

Adrenergic(1)

Contraction

Iris dilator

Adrenergic(1)

Contraction

Iris constrictor/ Ciliary

Ach(M3)

Constriction

Platelets

5HT(5HT2A)

Aggregation

Cell type

Activators

Effects

Cardiomyocytes

Adrenergic(1)

Positive ionotropic
effect

Hepatocyte

Adrenergic(1)

Glycogenolysis,
Gluconeogenesis

Adipocyte

Adrenergic(1)

Glycogenolysis,
Gluconeogenesis

Proximal
Convoluted tubule

Angiotensin II (AT1) Stimulate NHE3

Adrenergic (1)
H+ secretion & Na+
reabsorption
Stimulate
basolateral Na+-K+
ATPase
Na+ reabsorption

 IP3 Receptor
Ligand gated Ca2+ channel
High conc. in membrane of ER
Ligands which regulate:
1. PKA:
Ca2+ release by phosphorylation
2. PKG:
. Inhibits Ca2+ release
3. IP3:
. Ca2+ release
4. Ca2+:
. Conc of 100-300nM Ca2+ release
. Conc of 1000nMinhibits Ca2+
release
. Oscillatory pattern of Ca2+ release

 Ryanodine receptor
Present in skeltetal & cardiac muscles
& neurons
Major cellular mediator of
calcium-induced calcium release
Ca2+ enters through L-type Ca Channels
Conformational change in RyR receptors
Release of Ca2+ from SR into cytosol
Agonist: Xanthines (Caffeine, Pentoxyfylline)
Antagonist: Dantrolene

 Calcium Reuptake
Na+/Ca2+ exchanger on plasma membrane
Ca2+ pump on ER membrane
Ca2+ binding molecules
Ca2+ pump on Mitochondia

Effects of Ca2+ :
Cell Type

Effect

Secretory Cells (mostly)

secretion (vesicle fusion)

Juxtaglomerular cells

secretion

Parathyroid chief cells

secretion

Neurons

transmission (vesicle
fusion)

T-cells

activation in response to
antigen presentation

Myocytes

Contraction (TroponinC)

Calmodulin binds Ca2+ & activates 5 different

Calmodulin-dependent kinases
1. Myosin light-chain kinase Phosphorylates myosin
Contraction in smooth muscle
2. CaMK I synaptic function
3. CaMK II neurotransmitter secretion, transcription factor
regulation & glycogen metabolism
4. CaMK III protein synthesis
5. Calcineurin, a phosphatase that inactivates Ca2+ channels
by dephosphorylating prominent role in activating T cells
inhibited by some immunosuppressants

Therapeutic Applications:
Drug

Mechanism of
Action

IP3/DAG
Levels

Therapeutic
Application

Prazosin

1 blocker

Hypertension/
Prostate
Hyperplasia

Chlorpheniramine

H1 blocker

Allergies/
Common cold

Ipratropium
bromide

M3 blocker

Asthma/ COPD

Losartan

AT1 Receptor
blocker

Hypertension/
MI/ Diabetic
Nephropathy

Montelukast

LT C4/D4 blocker

Asthma

Oxytocin

Direct action on
Gq

Labour induction/
Uterine inertia

Cell surface receptors recruit activity of protein kinases in two general ways:
Receptor Tyrosine Kinases:
Possess an intrinsic tyrosine kinase activity that is part of the receptor protein
Examples include receptors for growth factors (PDGF, EGF, insulin, etc.)
Non-receptor tyrosine kinases:
Receptors lacking self-contained kinase function
recruit activities of intracellular protein kinases to the plasma membrane

Receptor Tyrosine Kinases

Implicated in diverse cellular responses:
Cell division, Differentiation & Motility
At least 50 RTKs identified:
Subdivided into 10 subclasses based on
differences within extracellular, ligand-binding domain of receptor

Structure:
Four common structural features shared
among RTKs
Extracellular ligand-binding domain
Single transmembrane domain
Cytoplasmic tyrosine kinase domain(s)
Regulatory domains

Receptor Dimerisation
Three ways in which signaling proteins can cross-link receptor
chains
1. Dimer ligand
2. Monomer but brought together by proteoglycan
3. Cluster on membrane

 Receptor dimerization leads to activation of

catalytic domains causing
autotransphosphorylation
Receptor autotransphosphorylation:
Further stimulates kinase activity
Leads to phosphorylation of additional
proteins involved in receptor
signalling pathway

Provides docking sites for downstream

signalling proteins
(Grb2, PI3-kinase, phospholipase C, etc.)
Src homology (SH) 2 & SH3 domains:
SH2 domains: bind P-Tyr-containing sequences
SH3 domains: bind to pro-rich (PxxP) sequences
Activates Phospholipase C
The binding of SH2-containing intracellular signaling
proteins to an activated PDGF receptor

RTK mediated pathways:

1. Ras-Raf-MAP kinase pathway
The activation of Ras by RTK signaling

The MAP-kinase regulated by Ras

Ras-Raf-MAP kinase pathway in Cancer

Ras gene mutation
defective Ras protein
GAP binds to GTP bound Ras, but
not able to provide domain for GTPase
GTP is not lysed & the GTP bound
Ras remains continuously active
Thus permanently activated MAP kinase pathway
results in growth factors transcription causing continuous cell
proliferation
Development of Cancer

2. PI3 Kinase Pathway

. Activated PI3 docks at the
phosphorylated RTK
. Brings about phosphorylation of
PIP2PIP3
. Downstream effects:
.
Inhibits proapoptotic protein BAX
.
Translation of tumour proteins by
activation of mTOR
.
Phosphorylation of FOXFO, antitumour protein,
ubiquitinisation degradation

Therapeutic Applications
I.

Receptor Tyrosine Kinase Inhibitors:

A. Epidermal Growth Factor Receptor Inhibitor/ HER 1 Inhibitor

. Geftinib:
.
Inhibits EGFR tyrosine kinase activity
.
Blocks ATP binding site
.
NSCLC pts. who have failed with std. chemotherapy
. Erlotinib
.
Similar mechanism of action
.
Locally advanced or metastatic NSCL & Pancreatic Ca

 Cetuximab:
Monoclonal antibody to extracellular domain of EGFR
Combined with Radiation for Locally advanced
Squamous Cell Ca of head & neck
EGFR positive metastatic Colorectal Ca
Panitumumab:
Recombinant fully humanized IgG to extracellular domain
of EGFR
EGFR positive metastatic Colorectal Ca

B. HER2/neu Inhibitor
. Lapatinib
.
Inhibits EGFR & HER2/neu Kinase activity
.
ATP binding pocket
.
Approved for Trastuzumab Refractory breast Ca with
Capecitabine
. Trastuzumab:
.
Humanized monoclonal Antibody to external domain
of HER2/neu receptor
.
Her2/neu overexpressing metastatic breast Ca
with Paclitaxel

II. mTOR inhibitors:

. IL-2 stimulates immune system by activation of T Cells via
activation of mTOR
Sirolimus (Rapamycin)
.
.
.
.

Binds to mTOR & inhibits action of IL-2

Immunosuppressive agent in organ transplant & GVHD
Cardiac stents to chances of re-occlusion
A/E: thrombocytopenia, hyperlipidemia, HUS

Everolimus
. Short t1/2
. Cardiac transplants

 JAK-STAT Receptor Pathway

Ligands: Interferon , Growth hormone, Prolactin
Receptors have no intrinsic activity
Intracellular domain binds intracellular tyrosine kinase
Janus Kinase (JAK)
Receptor mediated dimerization phosphorylation of
Signal transducers & activators of transcription (STAT)
STATs translocate to nucleus & regulate transcription
4 JAKs & 6 STATs combine differently depending on
Cell type & signal
Eg: Prolactin JAK1, JAK2 & STAT5

Therapeutic Application:
Lestaurtinib:

Janus Kinase 2 Inhibitor

JAK/STAT signaling exaggerated in MPNs
Polycythemia vera, essential thrombocythemia
& primary myelofibrosis
Mutant JAK2 activity
Inhibits wild type JAK2 kinase activity
Inhibits proliferation MPD cells
Phase II AML & Myeloproliferative disorders

 Receptor Serine-Threonine Kinases:

Anologous to RTK except they have
Serine/Threonine kinase domain in cytoplasmic region
Ligand: TGF
Dimerizes in presence of ligand
Phosphorylation of kinase domain activation
Phosphorylation of gene regulatory protein termed
Smad on serine residue
Dissociates from receptor associates with transcription factors
Morphogenesis & transformation
Inhibitory Smads: Smad6/7

 Toll like Receptors

Signaling related to innate immunity
Family of 10 receptors
Structure:
Single polypeptide chain
Large extracellular ligand binding domain
Short membrane spanning domain
Cytoplasmic TIR domain
Ligands: Pathogens (lipids, peptidoglycans, lipopeptides,
viruses)
Inflammatory response to pathogen

Signaling:
Receptor induced dimerisation
Recruitment of Mal & MyD88 to TIR
Recruits Interleukin-associated kinases
(IRAKs)
Auotphosphorylates & complex with MyD88
Also recruits TRAF6
Interact with protein kinase TAK1 & Adaptor TAB1
Activates NF-B
Trasncription of inflammatory genes

TNF Receptors
Structure:
Single membrane spanning receptor
Extracelluler ligand binding domain
Transmembrane domain
Cytoplasmic domain (death domain)
2 types: TNF1 (most cells) & TNF 2 (immune cells)
Activated by trimerisation

Has following effects:

Activation of NF-B:
Transcription of proteins involved in cell survival and
proliferation, inflammatory response & anti-apoptotic factors
Activation of the MAPK pathways:
The JNK pathway is involved in cell differentiation,
proliferation & is pro-apoptotic
Induction of death signalling:
Cell apoptosis

Therapeutic Application
Promotes inflammatory response & associated with
autoimmune disorders:
(Rheumatoid arthritis, ankylosing spondylitis, inflammatory
bowel disease, psoriasis, hidradenitis suppurativa and
refractory asthma)
Treated by using a TNF inhibitor:
Monoclonal antibodies such as
Infliximab, Adalimumab & certolizumab pegol
Circulating receptor fusion protein such as etanercept

Pharmacodynamic Interactions in a Multicellular

Context
Consider the vascular wall of an arteriole
Several cells interact at this site:
Smooth muscle cells(SMC), endotheial cells, platelets & postganglionic parasympathetic neurons
Effects produced
SMC contraction Ang II, NE
SMC relaxation NO, BNP, epinephrine
Alter gene expression PDGF, Ang II, NE, Eicosanoids

 Patient with hypertension

levels of AngII
activity of sympathetic nervous system
NO production
Pharmacotherapy directed towards:
BP
Prevent long term changes in vessel wall
Drugs used to treat hypertension
Ang II (Atenolol/Aliskiren/Enalapril/Losartan)
1 blockers NE binding on SMCs
NO production (Na Nitroprusside)

Conclusion:
Second messengers have shown to play physiological &
pathophysiological settings
Evolved as targets for drug develoment for numerous diseases
However, availability of compounds acting on specific targets is
the biggest challenge
Also is their difficult expression & purification
Such specific drugs are still in their early stages of development
Todays drug targets, tomorrows blockbusters