Nanosuspensions

Presented by : Mr. Satish S.

Reddi
Guided by : Dr. M. R.
Bhalekar

AISSMS COLLEGE OF PHARMACY, PUNE

 Content

 Introduction
 Methods of preparation
 Sterilization and stability aspects
 Characterization
 Applications
 Market status of nanosuspension
 References

05/28/10 2
 Introduction

 Nanosuspensions of drug are sub-micron colloidal

dispersions of pure particles of drug, which are
stabilized by surfactant.


 They are distinguished from polymer

nanoparticles, which are polymeric colloidal
carriers of drugs and from solid lipid
nanoparticles, which are lipidic carriers of
drugs.


 Nano is Greek word, which means ‘dwarf’. Nano is

the factor of 10-9 or one billionth.
 0.1 nm = Diameter of one hydrogen atom

05/28/10 2.5 nm = Width of a DNA molecule 3
 Why..?

W h y n a n o su sp e n sio n , w h e n o th e r
co n v e n tio n a l m e th o d s o f s o lu b ility
im p ro v e m e n t a re a v a ila b le . . ?

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 Decision tree for selection of
formulation approach
Co - solvent
Co - solventsInclusion
justment salt formation complexNanosuspension Emulsion
Other lipidic carri

Yes
Suitable molecular shape
No Low
Yes
Hig
h Dose?
Low
Melting point?
Hig
Low h
log P?
Water soluble? Hig
No h
Can salt be made?
Yes No

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Start 5
 Formulation consideration

 Stabilizers
 e.g. cellulosics, poloxamers, polysorbates,
lecithins and providones


 Organic solvents
 e.g. ethyl acetate, ethyl formate, butyl lactate,
triacetin, etc


 Co-surfactants
 e.g. bile salts, dipotassium glycerrhizinate,
transcutol, glycofurol, ethanol, isopropanol, etc


 Other additives

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e.g. Buffers, Osmogents, Antimicrobials,6
 Advantages of
nanosuspensions
 Increase in dissolution velocity:
 It can be explained using by Noyes Whitney
dissolution model equation
 Increase in saturation solubility:
 It is explained by Ostwald-Freundlich’s
equation
 Long term physical stability
 Ease of manufacture and scale up
 Versatility
 Improved biological performance…
05/28/10 7

 …Improved biological
performance
Route of administration Potential benefits
Oral Rapid onset of action
Increased bioavailability of drug
Reduced fed/fasted ratio
Intravenous Sustained release via monocyte phagocytic system
targeting
Reduced toxicity
Increased efficacy
Ocular High drug loading
Higher bioavailability
Pulmonary Increased delivery to deep lung
More consistent dosing

Subcutaneous/ Intramuscular Reduced volume of administration

Reduced irritation
Higher bioavailability

05/28/10 8
 Methods of preparation

 Bottom up technology
 Top down technology
 1. Pearl / Ball milling
 2. High Pressure Homogenization
 Melt emulsification method
 Emulsion or microemulsion as template
 Aerosol flow reactor method
 Supercritical fluid based technologies
 Highee technology

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 Bottom up technology

Drug Solvent

Drug solution

Antisolvent

Precipitation Nanosuspension

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 Bottom up technology
 It is also known as precipitation technique.
 It involves two steps:
 1. Initial creation of crystal nuclei
 2. Subsequent growth
 Particle size controlling parameters:
 1. Temperature
 2.Supersaturation
 3. Surfactant concentration
 Advantages
 Disadvantages
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 Pearl milling
Schematic representation of pearl
milling

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 Pearl milling
 Also known as wet milling
 Particle size controlling parameters:
 1. No. of cycles
 2. Speed of rotation
 3. Hardness of drug
 Dispersion with mean diameter >200nm is
obtained within 30-60 min
 Advantages
 Disadvantages

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High pressure homogenization
HPH ( APV micron LAB 60 )
It has following types:
 Microfluidisation
 Piston-gap homogenizers
 a. Dissocubes
 b. Nanopure
 c. Nanoedge


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 Microfluidisation

 Also known as MRT (Microfluidics Reaction

Technology).
 In consist of reaction chamber where precipitation
or crystallization takes place.
 Short dwell time inside high speed, high pressure
and high shear environment forces the reactant
to interact at nanoscale level.
 It consists of two reactors:
 1. Microfluidics mixer reactor (MMR)

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2. Coaxial feed reactor (Co-Reactor) 15
 Microfluidisation

Schematic representation of microfluidizer

Cooling jacket
Outlet
Inlet reservoir

Pump

Pressure
gauge

Patented interaction
chamber

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 Microfluidisation

 Particle size controlling factor:

 1. Feed rate
 2. Reactant ratios
 3. Mixing intensity
 4. Pressure
 5. Residence time
 Advantages
 Disadvantages

05/28/10 17
 Dissocubes

 This process is also known as Homogenization in

water
 Developed by R. H. Muller et. al. (1999)
 Instruments used:
 1. APV micron LAB 40
 2. APV micron LAB 60
 3. Avestin
 Instrument capacity: 40 ml t0 few thousand liters
 Pressure: 100 t0 1500 bars (2800 t0 23500 psig)
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 Dissocubes
Production of nanosuspension by HPH ( APV micron LAB 40 )

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 Dissocubes
Working principle of Dissocubes

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 Dissocubes

 Parameters controlling particle size:

 1. Power density of homogenizer
 2. Number of homogenization cycle
 3. Temperature
 Advantages
 Disadvantages

PCS data of azodicarbamide drug as function of

no . of homogenization cycle
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 Nanopure
 In this method suspension is homogenized in
water free media
 In Dissocubes technology:
 - cavitation is rate determining step
 - statics pressure is not be sufficient to
initiate cavitation
 - disintegration requires higher temperature
 - not suitable for thermolabile
 In nanopure non-aqueous media homogenized at
00c or even less
 Also called as “deep-freeze” homogenization
05/28/10 22
 Nanoedge

Drug Solvent

Drug solution

Antisolvent
High Pressure
Homogenization

Precipitation Nanosuspension

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 Nanoedge
 This technique is combination of precipitation
and homogenization technique.
 Depending on precipitation condition, particles
can be completely amorphous, partially
amorphous or completely crystalline.
 Second high energy addition step preserve size
range and converts all precipitated particles to
crystalline material.
 In this way major drawback of precipitation
technique such as crystal growth and long
05/28/10 term stability can be resolved by this 24
 Nanoedge

After precipitation and

Raw material before homogenization After homogenization

05/28/10 25
 Melt emulsification method

Drug melt Solvent

Primary emulsion

High Pressure Homogenization

Cooling
05/28/10 26
 Melt emulsification method

 Particle size controlling factors:

 1. Rate of cooling
 2. Concentration of drug
 3. Stabilizers used

fect of surfactant and drug concentration on particle size ( PCS data

05/28/10 27
 Emulsion as template

 Applicable for drugs which are soluble in organic

solvent or partially water miscible solvent
 This method is used for poorly water soluble and
poorly bioavailable anticancer drugs shows five
fold increase in dissolution rate than
commercial product(Trotta et al 2001)
 Two ways of fabricating drug nanosuspension:
 1. Solvent diffusion method
 2. Solvent evaporation method
05/28/10 28
 Solvent diffusion method

Drug Partially water miscible solvent

Drug solution Water Stabilizer

Emulsion

High Pressure Homogenization

Precipitation
Highly water miscible solv

Nanosuspension
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 Solvent evaporation method

Drug Organic solvent

Drug solution Water

Evaporation Emulsion

Precipitation Nanosuspension

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 Emulsion as template

 Particle size controlling factor:

 1. Intensity of mixing
 2. Stabilizers
 3. Water miscibility of solvent
 4. Rate of evaporation
 Advantages
 Disadvantages


05/28/10 31
 Aerosol flow reactor method

Drug Volatile solvent

Atomization using inert gas

Nanodroplets suspended in inert gas

Heated tubular laminar flow

Solvent evaporation and Precipitation

Nanosuspension
05/28/10 32
 Aerosol flow reactor

 Particle size controlling factors:

 1. Temperature
 2.Solution concentration
 3. Type of atomization and atomization
efficiency
 4. Drug solubility
 Advantages
 Disadvantages


05/28/10 33
 Other methods

 Supercritical fluid based technologies:

 1. Rapid expansion supercritical process (RESS)
 2. Rapid expansion from supercritical to aqueous
solution(RESAS)
 3. Supercritical anti-solvent process (SAS)
 4. Supercritical anti-solvent enhanced mass
transfer (SASEM)


 High gravity reactive precipitation (Highee

technology)
05/28/10 34
 Overview of technologies

Nanocrystal Company Patent / Patent

Hydrosol Novartis (prev. application
GB 22 69 536
NanomorphTM Sandoz)
Soligs / Abbott GB 22 00 048
NanoCrystalTM elan Nanosystem D 19637517
Dissocubes® SkyePharma US 5145684
Nanopure PharmaSol US 5858410
NANOEDGETM Baxter PCT/EP00/0635
Microfluidics® Microfluidics Inc US 6018080

05/28/10 35
 Sterilization aspects of
nanosuspension
 Thermal sterilization (Autoclaving):
 1. The physical stability depends on composition
of stabilizing surfactant or surfactant mixture.


PCS data after autoclaving ( 15 min , 2 bar , European Pharmacopeia )

Drug Surfactant PCS Diameter

Before After
9% RMPK 0.6 % Phospholipon autoclaving
380 nm autoclaving
402 nm
22
9% RMPK -90
0.3% Tween-80 382 nm 1039 nm
22
05/28/10 36
 Sterilization aspects of
nanosuspension
 Aggregation is due to reduced efficiency of steric
stabilizer.
 Increase in temperature causes dehydration
consequently decrease in thickness of steric
stabilizer layer.
 Some steric stabilizer even reach their critical
flocculation temperature (CFT) at autoclaving
conditions.
 To Summarize:
05/28/10 37
 Sterilization aspects of
nanosuspension
 Radiation sterilization (γ irradiation):

LD data after γ irradiation ( 25 kGy for comparison only )

Drug Surfactant LDdata (diameter 99 %)




Before After
9% RMPK 0.6 % Phospholipon irradiation
3.78 micron irradiation
3.70 micron
22
9% RMPK -90
0.3% Tween-80 4.35 micron 4.26 micron
22

§ Stability under γirradiation was observed for a

range of other drug nanosuspension.

05/28/10 38
 Sterilization aspects of
nanosuspension
 In contrast, tarazepide nanosuspension shown
distinct increase in size after γ irradiation.
 When measurement of zeta potential carried out
using Malvern Zetasizer at field strength 22
V/cm found that:
 zeta potential before γirradiation: - 30.5 mV
 zeta potential after γirradiation: - 22.3 mV
 Drop in zeta potential shows that some chemical
reaction must have taken place.
05/28/10 39
 Sterilization aspects of
nanosuspension
 Zeta potential of about ± 20 mV is not sufficiently
high to stabilize the particle long term.
 For long term stability zeta potential of about ± 30
mV is required.
 To summarize:
 γ irradiation is possible with some
nanosuspension; it depend on nature of
stabilizer and drug.

05/28/10 40
 Sterilization aspects of
nanosuspension
 Aseptic production:
 1.Baxter realized on aseptic line for production
of parentral nanosuspension.
 2. In addition, it should be kept in mind that
HPH process itself has germ reducing effect.
 3. Not only drug particles but also the bacteria
are disintegrated.


05/28/10 41
 Long term physical stability of
nanosuspension

 Physical insatiability in nanosuspension is

expected due to Ostwald ripening
 According to Ostwald-Freundlich equation
saturation solubility increases with the
decreasing particle size
 This effect is more pronounce for particle below 1
micron.
 Diffusion phenomenon due to particle size
difference
05/28/10 42
 Long term physical stability of
nanosuspension
 Clofazemine nanosuspension showed distinct
increase in PCS diameter and LD 99% after two
month storage.
 RMPK-22 nanosuspension stabilized with
phospholipon-90 or tween-80 had PCS
diameter of 564 nm after production and 575
nm after two years of storage.
 To summarize:
 Storage of drug nanoparticles as an aqueous
suspension is possible if the stabilizing
05/28/10 surfactant mixture is optimal. Crystal growth 43
 Dosage form based on
nanosuspension
 Topical formulations:
 e.g. Creams , Water free ointments, etc.
 Dosage form for administration in the mouth:
 e.g. Sublingual system, Buccal system
 Dosage forms for oral drug delivery to the GI
tract:
 e.g. Tablets, Capsules, Pellets, etc.
 Pulmonary delivery of nanosuspension
 Parentral administration of nanosuspension
05/28/10 44
 Characterization test

 Particle-size distribution:
 1. Photon correlation spectroscopy (PCS)
 2. Laser diffractometry (LD)
 3. Coulter counter
 Crystalline state and morphology:
 1. Scanning electron microscopy (SEM)
 2. Differential scanning colorimetry (DSC)
 3. X-ray analysis



05/28/10 45
 Characterization test

 Zeta potential:
 1. Electrophoresis
 2. Malvern zetasizer
 Dissolution velocity/ Saturation solubility:
 The method described in the
pharmacopoeia
 e.g. shaking experiment at different
temperature (40, 200 & 400)
 Surface hydrophilicity/ hydrophobicity:
 Hydrophobic interaction chromatography
05/28/10 46
 Characterization test
 Chemical test:
 1. Active ingredients
 2. Degradation products
 3. Moisture ( for lyophilized and solid
dosage forms)
 4. Preservative
 5. pH
 Other specific test according to dosage from:
 e.g. for parentral: Sterility, Pyrogenecity,

05/28/10
Syringeability, etc. 47
 Applications

 Danazol nanosuspensions (Liversidge et al, 1995):

 Bioavailability 82.3% (reduced inter-subject
variability) whereas conventional suspension was only
5.2%


 Atovaquone nanosuspension (Scholer et al., 2001):

 1.Poor bioavailability of 10-15% because of
dissolution rate limited
 absorption
 2.Oral nanosuspension increased bioavailability
 3. High adhesiveness of drug particles sticking on
biological surfaces & prolonged absorption time


 Naproxen nanosuspension (Liversidge et al., 1995):

 1. Severe gastric irritation
 2. Reduced gastric irritation when particle size
reduced
05/28/10 48
 Applications

 Ketoprofen nanosuspension (Remon et al., 2001):

 1. Incorporated into pellets to release the drug for
24 h
 2. This approach facilitate delivery of BCS Class IV
molecules


 Bupravaquone nanosuspension (Kayser, 2001):

 1. Cryptosporidium parvum – main pathogen
causing diarrhea in immunosuppressant HIV
patients
 2. Targeting the drug to the pathogen located in
the epithelial membrane gut wall is essential -
Increasing the time for the drug in the GI tract to
prolong the pharmacological window with regard to
the fast washing out during diarrhea
 3. Bupravaquone nanosuspensions – surface
05/28/10 49
modified mucoadhesive nanosuspension – prolonged
 Market status of
nanosuspensions
Drug Indication Drug delivery Pharma Route status
company company
Paclitaxel Anticancer American American Intravenous Phase III
BioScience Pharmaceutica
l Partners

Sirolimus Immunosuppr Elan Wyeth Oral Marketed

( RAPAMUNE® ) essant nanosystem
Aprepitant Anti - emetic Elan Merck Oral Marketed
( EMEND® ) nanosystem
Cytokine Crohn’s Elan Cytokine Oral Phase II
inhibitor disease nanosystem PharmaScience
Busulfan Anticancer SkyePharma Supergen Intrathecal Phase I

Fenofibrate Lipid SkyePharma Abbott Oral Marketed

(TriCor ® ) lowering
Insulin Diabetes BioSante Self-developed Oral Phase I

05/28/10 50
 Market status of
nanosuspensions
Drug Indication Drug delivery Pharma company Route status
company
Undisclosed Anti-infective Baxter Undisclosed Oral or Preclinical to
multiple NANODOSAGE Intravenous Phase II

Undisclosed Anticancer Baxter Undisclosed Oral or Preclinical to

NANODOSAGE Intravenous Phase II

Diagnostic AgentImaging agents Elan nanosystem Photogen Intravenous Phase I/II

Thymectacin Anticancer Elan nanosystem NewBiotics / IIex Intravenous Phase I/II

Oncology
Megsterol Appetite Elan nanosystemRAR Oral Marketed
acetate stimulant Pharmaceutic
(MEGACE ®ES ) al

Fenofibrate Lipid Elan nanosystemFirst Oral Marketed

(Triglide TM ) lowering Horizon
Pharmaceutic
al
05/28/10 51
 Summary

 Attractive drug delivery method for enhancing

solubility and bioavailability
 Large scale production and sterilization is
possible
 Can be administered using various routes like
oral, parenteral, ocular and pulmonary
 Oral nanosuspensions can be converted to
dosage forms like tablets or capsules
 Success is evident by increase in the
commercially available products of
05/28/10 52
 References

1. Rainbow B. E., Nanosuspensions in drug delivery,

Nature reviews Drug discovery, 3 (2004) 785-
796
2.
3. Patravale V. B., Date A. A. and Kulkarni R. M.,
Nanosuspensions: a promising drug delivery
strategy, Journal of Pharmacy and
Pharmacology, 56 (2004) 827-840
4.
5. Arunkumar N, Deccaraman M and Rani C,
Nanosuspension technology and its application
in drug delivery, Asian journal of
Pharmaceutics, July-Sept 2009, 168-173
6.
7. Muller R. H., Jacobs C., Kayser O.,
05/28/10
Nanosuspensions as particulate drug 53
 References

5. Arunkumar et al, Preparation and solid state

characterization of Atrovastatin
nanosuspension for enhanced solubility and
dissolution, International journal of pharmtech
research 1 (2009) 1725-1730
6.
7. Leversidge M. E., Leversidge G. G. and Cooper E.
R., Nanosizing: a formulation approach for
poorly-water –soluble compounds, European
journal of pharmaceutical science ,18 (2003)
113-120
8.
9. Rainbow B. E., Nanosuspension for parentral
delivery; Thassu D., Deleers M., and Pathak Y.
(Eds), Nanoparticulate drug delivery, Informa
05/28/10 54
 References

8. Keck C. M. and Muller R. H., Drug nanocrystals of

poorly soluble drugs produced by high
pressure homogenization, European journal of
Pharmaceutics and Biopharmaceutics 62
(2006) 3-16
9.
10.Jia L. Et al, Effect of Nanonization on Absorption
of 301029: Ex Vivo and In Vivo
Pharmacokinetic Correlations Determined by
Liquid Chromatography/Mass Spectrometry
Pharmaceutical Research, 19 (2002) 1091-
1097
11.
12.Kayser O. Nanosuspensions for the formulation
05/28/10 of aphidicolin to improve drug targeting 55
 References

11.Date A. A. and Patravale V. B., Current strategies

for engineering drug nanoparticles, Current
opinion in colloid & interface science 9 (2004)
222 – 235.
12.
13.Keck C. M. and Muller R. H., Drug nanocrystals
of poorly soluble drugs produced by high
pressure homogenization, European journal of
Pharmaceutics and Biopharmaceutics 62
(2006) 3 -16.
14.
15.Leversidge E. M. et al, Nanosizing: a formulation
approach for poorly water soluble compounds,
05/28/10 European journal of pharmaceutical sciences 56
  Thank you …

05/28/10 57