PH 514 : Nanopharmaceutics

Convener: Dr. M. S. Muthu

Associate Professor
Department of Pharmaceutical Engineering &
Technology

Indian Institute of Technology (BHU)

Varanasi
 PH 514: NANOPHARMACEUTICS
1. GENERAL
1.1. COURSE TITLE: NANOPHARMACEUTICS
1.2. COURSE NUMBER: DE.PH514
1.3. CONTACT HRS: 3-0-0
1.4. CREDITS: 9
1.5. SEMESTER OFFERED: I
1.6. PREREQUISITES: None
1.7. SYLLABUS COMMITTEE MEMBERS: Dr. M.S.Muthu (Convener), Dr. Ashish Kumar Agrawal

2. OBJECTIVE
The course is aimed to provide the students the current knowledge of design and formulation of nanopharmaceuticals.

3. COURSE CONTENTS
Unit I. Introduction to Nanotechnology: principles and applications. (3 L)
Unit II. Formulation techniques of nanoproducts. (4 L)
Unit III. Optimization, characterization and in vitro and in vivo evaluation of nanopharmaceuticals. (9 L)
Unit IV. Stability testing of nanopharmaceuticals. (2 L)
Unit V. Nanoformulations for various delivery routes: oral, parentral, pulmonary, inhalation, skin, ophthalmic, etc. ( 8L)
Unit VI. Nanoformulations for tuberculosis, HIV, cancers and diabetes and cosmetic use (4 L)
Unit VII. Applications of nanotechnology for targeted and theranostic delivery (7 L)
Unit VIII. Advantages and disadvantages of nanoformulations. (2 L)

4. READINGS
Xing-Jie Liang, Nanopharmaceutics, The Potential Application of Nanomaterials. World Scientific Publishing Co Pte Ltd; 1st Edition. 2012.
Recent literatures from high impact journals.

5. OUTCOME OF THE COURSE

After the completion of this course, the student will be able to demonstrate the importance of nanotechnology and nanoformulations in drug
delivery.
 Introduction to Nanomedicine:
Drug Delivery & Targeting

09/12/2020 4
 Nanotechnology
• The National Nanotechnology
Material Molecular initiative defines nanotechnology
Science Biology as “the understanding and control
of matters at dimensions of
roughly 1-100nm”
NANO
Engineering Medicine
TECHNOLOGY
• Technology that facilitates
fabrication of devices on nanoscale
i.e., in the range of 1-100nm.
Chemistry Physics
• Albert Frank defined nanotechnology as “it is the area
of science and technology where, dimensions and
tolerances are in the range of 0.1 to 100 nm”.

• For drug delivery purpose, particles are in

nanotechnology in the size range between 10-1000
nm.
 Nanomedicine

• It is defined as “the medical application of nanotechnology which

shows immense potential to solve problems in medicine” i.e., to
diagnose, treat and prevent disease at cellular and molecular level.

• Nanoparticles are colloidal particles in size range between 1 to 1000

nm, in which, diagnostic or therapeutic agents are adsorbed,
conjugated or encapsulated for diagnosis and treatment or for both.
 Types of Nanomedicine

 Capsular System: These are the systems in which the drug is confined
to a hallow cavity in another phase surrounded by unique polymer
membrane.

Nanocapsule
e.g., liposomes, nanostructured lipid carrier etc.

 Matrix System: These are the systems in which the drug is physically
and uniformly dispersed through out the polymer matrix (Nano
spheres) .
Nanomatrix
e.g., polymeric nanoparticles, solid lipid nanoparticles etc.
 BASEDON
BASED ONCHEMICAL
CHEMICALNATURE
NATURE
 Organic
nanoparticles
 Liposomes
Liposomes: Bilayer vesicles
enclosing aqueous
compartment that can be
produced using amphiphilic
phospholipids and
cholesterol.(size-400nm)
 Micelles
• Micelles: They are self-assembly
colloidal structure with
hydrophobic core and hydrophilic
shell. (size- lesser than 100nm)
 Dendrimers
• Dendrimers: Synthetic
nanomedicines that comprises as
a highly branded spherical
particle. (size-10-100nm)
 Nanotubes
• Nanotubes: Cylindrical structure
of layer of graphene sheets.
 Solid lipid nanoparticles
• Solid lipid nanoparticles: They
are made up of solid
hydrophobic lipid core, made
from biocompatible lipids. E.g.
Triglycerides (size- 100nm)
 Polymeric nanoparticles
• Polymeric nanoparticles: They are
prepared using biodegradable
polymers to provide biological
compatibility with lesser toxicity
(size -200nm)
 Inorganic
nanoparticles
 Gold nanoparticles
• Gold nanoparticles: commonly
synthesized by chemical treatment
of chloroauric acid.
 Quantum dots
• Quantum dots: Quantum dots
(QDs) are nanometer-sized
semiconductor crystals that glow
when stimulated by an external
source such as ultraviolet light .
 Upcoversion
• Lanthenides: When it is excited
with NIR radiation it gives
fluoroscense in visible range. eg.
Gd3+ Near infrared
980nm
Upconversion
Gd3+ Gd3+
 Super paramagnetic iron
oxide nanoparticles
• Super paramagnetic iron oxide
nanoparticles: SPIONPs composed
of ferric iron (Fe3+) and ferrous
iron (Fe2+). It is biocompatible and
used as enhance MR contrast Fe3O4
agent.
 Surface Modification
 Researchers found that upon intravenous administration (i.v.) of a
non-coated nanoparticles, particles are eliminated by reticulo-
endothelial system (RES) or immune system within seconds or
minutes.
 Non-coated nanoparticles may spontaneously get adsorbed with
plasma proteins (opsonization process) and these proteins are
capable of interacting with the plasma membrane receptors of
monocytes and macrophages, thus promoting particle recognition
leading to elimination from systemic circulation.
 To overcome this limitation, Gref. et al. (1994) has proposed coating
of nanoparticles with amphiphilic co-polymers in order to prolong the
blood circulation of nanoparticles after Intravenous administration.
 OPSONIZATION PROCESS

A.
Binding
Uncoated NP
Uncoated NP Opsonization
Oops

Complement fixation and cellular uptake

B.

Not Binding
PEGylated NP Repulsive forces
No Opsonization
PEGylated NP
 These polymers should be inert, well solvated and compatible with the
solvent, and have polarizability close to water.

 This process of coating is known as PEGylation and PEGylated particles

ensure long circulation times, effective transport across the tumor
vessel wall and deep penetration into the tumor interstitium.
 Some of the commonly used Amphiphilic Copolymers for surface modification are

1. Brushed Poly Ethyleneglycol (bPEG)

2. Poly Ethyleneglycol (PEG)
3. Poly Ethyl Ethylene Phosphate (PEEP)
4. Poly–2–ethyl–2–oxazoline (PEtOx)
5. Poly Vinyl Pyrrolidone (PVP)
6. Dextran
 Surface Modification

Uncoated Nanoparticles Coated Nanoparticles

B.
A.
IONP C.
IONP AuNP

SLN
AuNP SLN

D. E.

Liposome UCN UCN

AgNP
Liposome
F.
AgNP
AgNP

(A) bPEG (B) PEG (C) PEEP (D) PEtOx (E) PVP (F) Dextran
 Enhanced Permeation and Retention (EPR) Effect

 Generally 10 – 100 nm sized particles begin to accumulate within the

tumors due to hyper and leaky vascularization, leaky vasculature and
ineffective lymphatic drainage in tumors leading to the enhancement
of therapeutic activity of drug containing nanoparticles.
 The vasculature in tumor is hyper permeable and this hyper
permeability is due to large intercellular openings between the
endothelial cells that comprise the neoplastic vessel wall.
 This process of size specific accumulation as a result of hyper
permeable vascularization is known as Enhanced Permeability and
Retention (EPR) effect.
 Targeting Methods
• Targeting active molecules to specific sites in the body had been
pursued actively ever since Paul Ehrlich first envisaged the use of ‘magic
bullets’ for the therapy of various diseases.

• Interest in this concept has increased significantly in recent decades

with the innovations of nanomedicine.

• Targeted delivery can be achieved by either passive or active targeting.

 Passive Targeting
 Passive targeting is achieved by loading drug into a nanocarrier that
reaches the target organ passively.
 Ex- passive targeting of tumors takes advantage of hyper-permeable
cells owing to their rapid vascularization.
 This rapid vascularization results in leaky, defective cells and impaired
lymphatic drainage.
 Nanoparticles ranging from 10 to 100 nm then begin to accumulate
within tumors because of their ineffective lymphatic drainage.
 This results in a phenomenon known as the enhanced permeation and
retention (EPR) effect.
 Nanoparticle size must be less than 200 nm to avoid uptake by the RES
and its surface should be hydrophilic to avoid clearance by
macrophages.
 Tumor vasculature:
Endothelial cells
with large pores
Angiogenic
blood vessels

Tumor

Blood
vessel

Normal vasculature:
Endothelial cells Extravasation of
with tight junctions Lymphatic
circulating Improper vessel
nanoparticles within lymphatic
the tumour due to EPR drainage
effect

Tumor vasculature showing EPR effect (Particles start accumulating)

 Tumor vasculature:
Endothelial cells
with large pores
Angiogenic
blood vessels

Tumor

Blood
vessel

Normal vasculature:
Endothelial cells Extravasation of
with tight junctions Lymphatic
circulating Improper vessel
nanoparticles within lymphatic
the tumor due to EPR drainage
effect

Tumor vasculature showing EPR effect (particles accumulated)

 Active Targeting
 Active targeting of a drug is achieved by conjugating a nanocarrier system
(drug loaded) to a tissue- or cell-specific targeting ligand.
 Active targeting has raised the importance of nanomedicine and this can
now be achieved by a number of specific interactions, such as ligand–
receptor and antibody–antigen binding.
 These specific interactions result in preferential accumulation of
nanomedicine into molecular targets.
 The different targeting ligands such as antibodies (Herceptin, Mabthera
and Erbitux , are antibodies that recognize HER2, CD20 and EGFR receptor,
respectively), small molecules (folic acid, whose receptor expressed on the
surface of the tumor cells) or peptides (amino acid sequence [Arg-Gly-
Asp], which bind to tumor αv β3 integrin) are attached covalently to the
surface of nanoparticles in the nanomedicine for active targeting of
anticancer drug to the malignant cells.
 Mechanisms of Cellular Uptake
1. Simple Diffusion 2. Non Receptor-Mediated 3.Receptor-Mediated Endocytosis
Endocytosis

Receptor recycling

1. Simple Diffusion – Low concentration of drug reaches to the target.

2. Non-Receptor-Mediated Endocytosis – Medium concentration of drug reaches to the target.

3. Receptor-Mediated Endocytosis – High concentration of drug reaches to the target.

 Advantages of Targeted Drug
Delivery to Cancer

1. Cancer chemotherapies have dose related side effects owing to

nonspecific biodistribution of drugs.

2. Cancer targeted delivery improve the therapeutic index of drugs by

preferential localization at target sites, lower distribution in healthy
tissues.
 PREPARATION OF
NANOPARTICLES BY SOLVENT
EVAPORATION TECHNIQUE
 Methods
1. Solvent evaporation
2. Double emulsification
3. Emulsions- diffusion method
4. Nanoprecipitation
5. Coacervation
6. Salting out
7. Dialysis
8. Supercritical fluid technology
Solvent Evaporation Technique

Aqueous phase

Organic phase
Drug Surfactant Polymer
 Solvent Evaporation Technique

Sonication Evaporation
Aqueous phase

Organic phase

Nanoparticle suspension

Drug Surfactant Polymer

CHARACTERIZATION OF NANOPARTICLES
1.Particle size and polydispersity index
2.Zeta potential
3.Drug entrapment/drug encapsulation
4.Drug loading
5.Shape and morphology
6.In-vitro drug release
 1. Particle
size
analysis

Microscopical Dynamic
light
scattering

2D 3D

Scanning Transmission
Atomic force
electron electron
microscopy
microscope microscope
 A
Dynamic light
scattering Laser

Intensity
Time
B
Measures
particle size and Laser

Intensity
its distribution

Time

Fig: Hypothetical fluctuation of scattered light in dynamic light scattering from (A) large and (B) small particles. As
larger particles generates higher scattering intensity than smaller particles 42
Polydispersity index
• Follow Contin Algorithm

 
PI = (width/mean)2

PI where D(0.9)= correspond to particle size immediately above

90% of the sample
D(0.1)= correspond to particle size immediately above
10% of the sample
D(0.5)= correspond to particle size immediately above
50% of the sample
Value suggested to define dispersity :
< 0.1 – highly monodisperse
0.1-0.4 – moderately polydisperse
> 0.4 – highly polydisperse
2. Zeta potential
• Define as electrical potential at the slip plane (meet of stern layer &
diffuse layer is slip plane layer)

• Stability of nanoparticle on the basis of charge:

± 0-10mv – highly unstable

± 10-20mv – moderately stable
± 20-30mv – highly stable
3. Drug Entrapment / Drug
Encapsulation
Entrapment used Encapsulation used
for matrix system in ENCAPSULATION
for capsular system
ENTRAPMENT contain inner void
which drug is
equally distributed space in which drug
in polymers is filled
 
% Entrapment

x 100

 
% Encapsulation

x 100
4. Drug loading

•%  Drug loading

x 100
 5. Shape and Morphology

Scanning Transmission Atomic

electron electron force
microscopy microscopy(T microscopy
(SEM) EM) (AFM)

Surface
Size Aggregation morphology
Size, Shape Heterogenicity
Distribution
6. In-vitro drug release
Methods:

Membrane diffusion method

Sample separation method

Continuous flow method

 In- vitro drug release study for nanoparticles

Dialysis bag diffusion Technique

Dialysis bag diffusion Technique is the one of the most
convenient method to determine the drug release profile of
the nano-formulations like nanoparticles, liposomes, micelles
and is based on the principle of diffusion of drug against
concentration gradient through a semipermeable membrane
until equilibrium is reached. To determined the amount of
drug release samples are
 Dialysis bag diffusion Technique

Empty Dialysis bag Dialysis bag with

nanoparticle suspension

In- Vitro drug release

 In-vitro drug release mechanisms

 Depending on the mechanism of drug release from the formulations,

it can be divided into four categories:

1. Rapid release/ burst release: (desorption of the surface

bound/ adsorbed drug)
2. Diffusion of drugs distributed in matrix.
3. Erosion of matrix.
4. Diffusion and erosion both.
In-Vitro drug release mechanisms from Nano-formulations

Drug adsorbed
1. Rapid release
Polymer

2. Diffusion of drugs (10-500 nm)

distributed in
matrix
Drug distributed

3. Erosion of matrix

Time = 0
4. Diffusion and erosion both
Advantages of Nano Drug Delivery and Targeting

 Enhancement of bioavailability
 Improves stability
 Enhancement of therapeutic efficacy/index
 Enhancement of solubility
 Personalized treatment - Theranostics
 Reduction in systemic side effects of drugs
 Targeted smart delivery
 Enable controlled release
 Reduced morbidity and mortality rates
 Disadvantages of Nanomedicine

 High Cost

 Variation in receptor overexpression

 Difficulties in scale-up