PACLITAXEL

New Parenteral
Drug Formulation

Daniel Daniel Sanna Sirajuddin Shantal Wijesuriya Barbara Khoo

Acknowledgements
We would like to thank Dr. Roldo, Dr Ahmad and
Dr. Van Der Merwe for their guidance and
invaluable help towards this presentation.

Aim
To propose a more suitable drug delivery system
(DDS) than the current available one for
paclitaxel that improves on its therapeutic
profile in reducing its toxicity.

To reduce the sensitivity reactions and

intolerability with Cremophor-based paclitaxel
formulations.

Paclitaxel Discovery

1962

1964 - 1965

e 1: Paclitaxel discovery; timeline and geographical relevance

Introduction to Paclitaxel

Derived from the bark of taxus brevifolia

[1]
Paclitaxel is a mitotic / microtubule
inhibitor.
Highly lipophilic and insoluble in water
Treatment for various types of cancer
such as ovarian cancer, breast cancer
and non small cell lung cancer. [2]
How does paclitaxel exert its
effects?
It inhibits the completion of cell division
by disrupting microtubules [5]
gure 2 = Chemical structure of paclitaxel [3]
It enhances the polymerization of
tubulin to stable microtubules.
Paclitaxel Properties:[4]
Paclitaxel specifically binds to the beta
subunit of tubulin, blocking cells in the
White to off-white crystalline
G2/M phase of the cell cycle and as a
powder
result the cell is to undergo mitosis. [6]
Empirical formulaC47H51NO14
Molecular weight-853.9 Da

Liver Cancer
Liver - one of the major
bodily organs
Functions :
Digestion of proteins and fats
Removal of toxins from the
body
Clotting factor production
Symptoms
experienced in the
Bile production
advanced stage:
Unexplained weight loss
Loss of appetite
Nausea and vomiting
Swelling of your abdomen
(tummy)
Jaundice
Fatigue

Liver cancer - a disease

that involves the
uncontrolled proliferation of
cells.
It can begin either as:
Single tumour grows and
only in the late stage spreads
to other parts of the liver.
Many spots throughout the
liver this is usually due to
cirrhosis (caused by chronic
hepatitis or long term alcohol
use).

Current formulations
available
FDA have approved Taxol for the first line treatment of
ovarian and breast cancer. To this date, Taxol is widely
used as an anti-cancer drug and also in other combination
therapy regimens 1.

Current administration:
IV Continuous infusion
IV Intermittent infusion

Problems with current formulation:

The vehicle in which paclitaxel is formulated in, Cremophor
EL, causes hypersensitivity reactions
DHEP leaching
Precipitation of paclitaxel

Nanoparticles for solid

tumor delivery[1]
Effectiveness depends on the adequate delivery of the
therapeutic agent to the tumor cells.

Nanoparticles would overcome the lack of specificity of

conventional drugs which have minimal selectivity for
malignant cells.

This minimal selectivity causes high toxicities against

rapidly dividing normal cells.

Nanoparticles can avoid rapid clearance by the

reticuloendothelial system and can accumulate in solid
tumors.

Increased stability for storage and in biological fluids

Rationale behind our drug delivery system

Advanced parenteral
drug delivery system

Defining objectives of
study and planning

Screening of factors and

literature studies

Drug:
Paclitaxel

Passive
Liver
EPR effect

Specificity of drug
delivery system

Targeting cells

Activ
e
Biomaterial

Galactosami
ne

Micelles
Liposomes

Lipid based
nanoparticles to
formulate anti-cancer
drugs

Colloidal carriers

Nanoemulsi
ons
Nanocrystal
s

IV

IM / SC

Parenteral
Administration

Micelles
Why Micelles?

Target specific tissues via passive or

active targeting.

Enable prolonged circulation and drug

retention

Large enough particle size to avoid renal

excretion

Small enough particle size to bypass

filtration by interendothelial cell slits in
the spleen

PEG PCL Block Copolymer

Polycaprolactone (PCL)

Hydrophobic segment, biodegradable, biocompatible and

nontoxic thermoplastic polyester (1)
Polyethylene Glycol (PEG)

Hydrophilic segment reduce the adhesion of plasma proteins,

soluble in water and organic solvents, stabilize particles, nontoxic, potentially increase the circulation time of drugs and
can
prevent
recognition
by
macrophages
of
the
reticuloendothelial system (RES) after intravenous injection.
(1)

The combination of Both polymers forming an amphiphillic

polymer capable of self assembly to form a core shell that can
act as a reservoir for paclitaxel

Formulation
Considerations
Other Additives

Form stable nanosuspensions with copolymers

Lysine and phenylalanine or lysine and leucine
[2]

Stabilizer

Retard ostwald ripening and produces a high

physical stable formulation [1]
Block Copolymer

gure 3 = Table of Nanosuspension formulation considerations

Paclitaxel Loaded Block Copolymers

The basis for the use of Block copolymers

They have been selected for their potential in solubilizing hydrophobic

drugs, reducing drug toxicity, and extending drug circulation in vivo. [5]

In order to avoid Ostwald- ripening and recrystallization with Paclitaxel,

stabilizing the formulation requires the conjugation of a hydrophobic
affixing marker that reduces its solubility and provides a more controlled
drug release rate. [5] It will ensure the containment of the solute in the
nanoparticle core whilst avoiding enzymatic activities.

Figure 4 = Plasma
Concentration time curve
of PTX loaded polymeric
micelles in comparison to
Taxol after IV
administration to rats [1]

Preparation of poly(ecaprolactone)-poly(ethylene
glycol) block co-polymers
Ring opening polymerisation[1]

PCL-PEG block co-polymers are synthesised by ring-opening

polymerisation (ROP) of -caprolactone in the presence of the
catalyst stannous octanoate diluted in dried toluene.

The solution is then heated at 130C under reduced pressure to

remove toluene and purified with dry nitrogen gas.
Polyethylen
e glycol
(PEG)[2]
Stannous Octanoate at
130C

ecaprolacton
e (CL)

Poly(e-caprolactone)
poly(ethylene glycol) (PCL-PEG)3

Formation of PTX loaded PCLPEG block copolymer

Emulsion/solvent evaporation technique

The block copolymers are dissolved in ethyl acetate (used instead of

methylene chloride or chloroform due to their safe environmental use and
decreased human safety concerns). [2]

Drug added: paclitaxel

Once dissolved they undergo vortex and are emulsified in 0.1wt.% sodium
cholate solution

The solvent evaporates in a vacuum oven at 37C for 1 hour

Suspension is filtered through 0.8mm membrane filter
Centrifugation is carried out for 60 minutes at 4C to recover the
nanoparticles.

The supernatant was discarded and the phosphate buffered saline (pH
7.4) is used to re-suspend the nanoparticles .

Micelle Assembly

Figure 5 = The assembly of the PTX loaded polymeric nanoparticle [1]

Conjugation of galactosamine to
paclitaxel block copolymer
Conjugation of ligand

Galactosamine is conjugated to the surface of nanoparticles via an

amide linkage by the use of activating reagents EDC and NHS.

Ultrafiltration

separates the nanoparticles

molecules and then they are lyophilised.

from

unreacted

Rationale
Attachment of a targeting
ligand enables the drug to
actively target a specific
tumour cell. Research shows
that Gal-P/NPs had significant
efficacy in the reduction of
the size of the tumour. This is
because a large number of
the Gal-P/NPs actively
targeted at the tumour site

Figure 6 = the conjugation of

galactosamine to PTX loaded block
copolymer used as a targeting
molecule.

Nanosuspension
The paclitaxel nanosuspension formulation is best suited
for parenteral administration [3] and have been found
to increase the efficacy of the parenterally administered
drug [4]

Nanosuspensions are colloidal dispersions of nano- sized

drug particles that are stabilized by surfactants [1]

The desired formulation requires our paclitaxel/PEG-PCL

block
copolymer
(surfactant)
to
stabilize
the
nanosuspension and is a major determining factor in
improving the formulation bioavailability [5].

Nanosuspensions allow the delivery of:

Poorly water-soluble drugs and Poorly lipid-soluble drugs[1]

Administration
Aseptic preparation of
nanosuspension for
administration
- Aseptic conditions
Storage:
- Non-PVC infusion
bags
- Photosensitivty
Continuous infusion
- Short half life
- Polyethylene lined
sets and a
catheter.

Figure 7 = Various routes

of parenteral
administration [1]

IV is the chosen route of

administration as
Paclitaxel is poorly soluble.
Rapid onset of action
Reduced dose
Target drug to site of
action

Administration animation
= Micelles

Receptor

Nucleus

Blood Vessel

Nanosuspension
I.V. Injection

Normal vessels
have a tight
endothelium

Tumor

Administration animation
= Micelles
Blood Vessel

Receptor

Nucleus

Active Targeting
Passive Targeting

Nanosuspension
I.V. Injection

Normal vessels
have a tight
endothelium

Tumor

Angiogenic tumor
vessels are leaky and
permeable

Nanoparticle pathway

Figure 8 = PTX loaded Block Copolymer in Cancer Tissue

cells [1]

Cancer cell uptake of drug

loaded nanoparticle
Hepatocytes recognise galactose and N-acetylgalactosamine- terminated
glycoproteins via the asialoglycoprotein (ASGP) receptor.

The ligand galactosamine is used to ACTIVELY target these ASGP receptors.

By the process of endocytosis the ligand-receptor complex is taken up by the
hepatocytes Endolysosomal release occurs and releases the PTX. The
receptor recycles back to the surface of hepatocytes.

Figure 9 = Active
Targeting pathway of
the administered drug
loaded nanoparticle

Summary of Adv vs.

Limitations
Advantages

Limitations and
problems

Reduce risk of toxicity effects and

sensitive reactions
high dose of paclitaxel could be
administered without any premedication
with steroid and antihistamines (typically
required to alleviate Cremophor related
hypersensitivity)[1].
Overcoming the solubility problems of
poorly soluble paclitaxel
Increase drug circulation time
Protecting the drugs bioactivity and
stability
Higher Affinity for receptors / avoiding
toxicity
Avoiding
rapid
clearance
by
the
reticuloendothelial
system
thereby
facilitating the reduction of tumor growth
Overcoming the multidrug resistance

Adverse side effects in

the body can be induced
by side products formed
during synthesise or by
polymer itself or by that
lead to hyper-sensitivity
Release of acidic
degradation products
Long term health related
issues of excipients are
unknown
Specificity to liver cancer

Future
Development
Magnetic nanoparticles as carriers in anticancer-drug delivery
The addition of nano-magnets to our delivery system?
- to overcome non-specific, improve the efficacy and reduce side
effects of chemotherapy.
Advantages : high responsibility in magnetic field, excellent
biocompatibility, long circulating time in vivo and relative stability
to guarantee the targeted delivery and delayed release of
anticancer drug.
Figure 10 = Various Nanomagnet Nanoparticles
After intravascular injection,
magnetic nanoparticles can be
transported by the blood
circulation and concentrated at
the tumor with the aid of a
magnetic field applied at the
affected zone. Removal of the
carrier by magnetic separation
after the drug nanoparticles

ure 11 = The use of nano-magnets in anti-cancer therapies

Summary
The limitations of the current formulation of
paclitaxel were overcome via the use of
paclitaxel polymeric micelles. (insolubility of PTX)

In conclusion the Gal-P/NPs can effectively target

the site of hepatoma tumour via the ASGP
receptor mediated recognition and significantly
reduce its size.

Therefore, the prepared Gal-P/NPs may be used

as a potential drug delivery system for the
targeted delivery to liver cancers.

Thank you
Thank you for listening
Do you have any questions?

References