PREINITIATION

INITIATION

PROMOTION

PROGRESSION

MALIGNANCY

exposure to
cancerogenes

mutations

cumulations of
mutations

cancer in situ

metastasis

CELL CYCLE NON SPECYFIC AGENTS (CCNSA)

ALKYLATING AGENTS

CYTOTOXIC ANTIBIOTICS

PLATINUM COMPOUNDS

NATURAL PRODUCTS

CELL CYCLE SPECYFIC AGENTS (CCSA)

G2

19%
(Pre-mitosis)

ANTIBIOTICS - BLEOMYCIN

40%
(DNA - synthesis)
ANTIMETABOLITES
ANTIFOLIATES

G1

39%
(Pre-synthesis)

2%
(Mitosis)

PLANT ALKALOIDS

G0

(Resting)

O
CH3

(a) Diethylpropion HCl (1-phenyl-2-diethylamino-1-propanone hydrochloride)

CH3

CH3
CH 3

(b) Phentermine ( , , dimethylphenethylamine hydrochloride)

(c) Topiramate (2,3:4,5-Di-Oisopropylidene- -D-fructopyranose

sulfamate)

HCl

CH2 C

HCl

NH 2

CH 3

CH2 OSO2NH2

O
H3C
H3C

O N

(d) Zonisamide (benzo[d]isoxazol-3ylmethanesulfonamide)

CH3

CH3

O
S NH
2
O

H
O

(e) Orlistat ((S)-2-formylamino-4-methylpentanoic acid (S)-1-[[(2S, 3S)-3-hexyl4-oxo-2-oxetanyl] methyl]-dodecyl ester)

H
O

(f) Rimonabant (5-(4-chlorophenyl)-1-(2,4dichloro-phenyl)-4-methyl-N(piperidine-1-yl)-1H-pyrazole-3carboxamide)

Cl

N N
H

N N

Cl
Cl

(g) Sibutramine Hydrochloride

(cyclobutanemethanamine, 1-(4chlorophenyl)-N,N-dimethyl- -(2methylpropyl)-,hydrocholoride)

CH3

H2C
Cl

CH3
N

CH3
CH3

HCl H2O

Figure 1:
extracellular space
GSH

GSSG GS-X

AA

-Glu-AA
Gly

Cys-Gly

GS-X/GSSG
transporter

GSH
transporter

-GT

Cys

DPD

AA
transporter

-Glu-AA

GS-X

GST
Pr-SSG

Pr-SH

Grx
GST

-GCT
AA

GPx

H2 O 2
LOOH

GSH
ADP

5-Oxoprolin

GSH-S
Gly

H2O+O 2

GSSG

GR

ATP

-Glu-Cys

Glu
-GCS

NADP+

cytosolic compartment

NADPH

5-OP
Cys

ADP

ATP

A.
lumen
cytoplasm

DHHC-CRD

B.
TM1 TM2

TM3 TM4

DPG
N-Variable

DHHC-CRD

TTxE
C-Variable

A
palmitate

O
S

H2N

N
H O

H
N

COOH

cysteine residue

B
azido-fatty acids
N3

azido-fatty acid
modified protein(s)

O
OH

live cells
purified proteins

biotin

N
H

O
S

N3

phosphine-biotin
Staudinger ligation
O O
PPh2
N
H
O

S-palmitoylation
O
S

biotinylated proteins

Streptavidin blot

affinity purification

mass spec analysis

2-bromopalmitate

tunicamycin

cerulenin

O
O

SH

Cysteine

+
O

CoA

Palmitoyl CoA

Protein
Palmitoyl
Acyltransferase

CoA
O

N
O

SH

S
N
Protein

Palmitoylated protein

Charles J. Malemud: Suppression of Pro-Inflammatory Cytokines via Targeting of

STAT-Responsive Genes
STAT
STAT
Responsive
Activator1
Cytokine/Protein
IL-2R

IL-10

IL-18R1

Activated
STAT(s)2

IL-4

STAT6

IL-2, IL-3

STAT5,
STAT5A,
STAT5B

IL-4

Major
Function(s) of
STATResponsive
Genes3
Complexes
with IL2R /IL2R
high
affinity IL-2R

Antiinflammatory
Cytokine

Activator of
JAK3, I-SRE4/IL-10 gene

T-Cell
Growth
T-Cell
Development

STAT6
Treg Cell
Development
IL18/IL-18R1

IL-6

IL-6ST(gp130)4

IL-6/IL-17
IL-3
IL-12

STAT3/STAT1
STAT5
STAT4

IL-19
IL-10/IL13

STAT3
STAT4

IL-6/IL17/OSM

STAT3/STAT4/STAT5A

IL-4

IL-2

INF-

IFN-

IL-3
IL-2
IL-12

Involvement
of STAT
activator in
RA

Activator of
JAK3

Representative
Reference

[81]

[73]
[207]

Inducer of
TNF- , GMCSF, IFNPromotes
Th17 Cell
Production;

MMP
Synthesis
MMP
Synthesis
Promotes
Th17 Cell
Production

Heterodimer
between
gp130/LIFR5
forms the
OSMR6
Th2
STAT3/STAT4/STAT5A
Differentiation
STAT1/STAT4/STAT5/STAT6
Activator of
Inhibitor of
multiple
Antiprotein
Inflammatory
kinases
Cytokine IL-4
and IL-10
Production

[127]

[9]
[174]
[140]

[207]

[140]
[100]

[118]
[86]

[207]
[207]
[113]

STAT5
STAT5
STAT4
STAT5
STAT3

Activator of
JAK2

OSM

IL-2/IL-3

TNF-

IL-3
IL-6/IL-19

STAT5
STAT3/STAT5

Activator of
JAK3;

IL-15
IL-22

STAT1/STAT3/STAT5

Activator of
p387, JNK8

Monocyte
Trafficking
MMP-2
VEGF
Activator of
NF- B

[122]
[125]

[200]
[86]
[186]

R
R

cells

Tight junction

Figure 2. Arrangement of epithelial cells with tight junctions

7.3. Cerebrospinal fluid barrier (CSF)

Epithelial cells which are in contact with the brain ventricular spaces form a barrier to the movement of drugs. These epithelial cells
are connected by occluding zonulae (blood- brain barrier) as shown in Figure 3. The zonulae severely restrict the passage of most
molecules between the bloodstream and the parenchyma of the central nervous system. Drug entry across this barrier is through
either passive diffusion or carrier mediated transport.Only the lipid soluble drugs cross into the CSF from blood.

Figure 3. Epithelial cells with tight junctions as part of the blood brain barrier

Epithelial cells that separate the CSF from the brain are connected with tight junctions and are characterized by marked scarcity of
pinocytic vesicles. However, the epithelial cells that lines the brain are not connected by occluding zonulae and therefore, there is
unrestricted passage of drug molecules from CSF to the brain. Drugs like penicillin which are not much lipid-soluble and required
in high concentrations for the treatment of brain abscesses are administered through intrathecal injections directly into the CFS.

7.4. Placental barrier

The placental membrane limits the amount of maternal blood following through the placenta to the foetus and passive diffusion is
the main mechanism of drug entry from the maternal blood to the foetus. The shortest time required for equilibration of a drug
between mother and foetus is about ten minutes and this delay is useful as it can allow a mother to be anaesthetized during final
stages of labour.

8. Systemic availability of drugs

A drug will reach systemic arterial circulation only if it is absorbed from the GIT and if it escapes metabolism in the gut, liver, and
lungs. When the concentration of the drug in plasma is measured at specified time intervals, it is possible to construct
concentration versus time graph and hence be able to determine the extent of drug availabilityas shown in Figure 4.
The availability depends on both the extent of absorption and the extent of presystemic metabolism and comprises three aspects;
Peak concentration (Cmax), Time taken to reach the peak (Tmax) and area under the curve (AUC)as shown in Figure 4. The Cmax and
Tmax are measures of the rate of availability while AUC is a measure of the extent of availability (i.e. proportion of the administered

Phase II clinical trials include inert placebos as negative controls and older active drugs as
positive controls alongside the investigative compound. These studies are done in special clinical
centers such as University Hospitals. A broader range of toxicities may be detected at this phase.
Phase IIIclinical trials
The drug is evaluated in a much larger number of patients (thousands) to further establish safety
and efficacy. Phase III trials are performed in settings similar to those anticipated for the
ultimate use of the drug. After successful phase III trials, the next step is the application for
review of the new drug to seek approval to use the drug for clinical management of the disease
condition.
Phase IV clinical trials
This phase is concerned withpost-marketing surveillance and the main goal is to assess adverse
reactions, patterns of drug utilization, discovery of additional indications.The interrelationships
between the various studies in drug development are illustrated in Fig 1.13 below;
Crude drug preparation

Biochemical profiling of
crude drug product

In vivo/in vitro
therapeutic profile for
test and control model

Phase IV
Clinical trials

Phase III
Clinical trials

In vivo toxicity profiles

for test and control
model

Phase I
Clinical trials

Phase II
Clinical trials

Figure 1.13Illustration of the key steps in the development of a drug from a putative drug
candidate extract

Prescribed dose
Patience compliance
Medication errors

Administered dose
Rate and extent of
absorption
Distribution and
composition of body fluids
and body size

Concentration at site of action

Physiological variables
Pathological and Genetic
factors
Interaction with other
drugs
Development of tolerance

Intensity of effect
Drug - receptor
interactions
Signal transduction

Figure 1.14: The operational levels that determine the relationship between prescribed d
dosage and the drug effect.

Drugs that are excreted primarily unchanged by the kidneys tend to have low variation am
patients with similar renal function than do drugs which are inactivated by metabolism. For
extensively metabolized drugs, those with high metabolic clearance and large first p
elimination have marked difference in bioavailability, whereas those with low biotransforma
tend to have largest variation in elimination rates among individuals.