REVIEWS

NANOSUSPENSIONS IN DRUG
DELIVERY
Barrett E. Rabinow
Abstract | A surprisingly large proportion of new drug candidates emerging from drug
discovery programmes are water insoluble, and therefore poorly bioavailable, leading to
abandoned development efforts. These so-called ‘brickdust’ candidates can now be rescued
by formulating them into crystalline nanosuspensions. In the process of overcoming issues
involving solubility, additional pharmacokinetic benefits of the drugs so formulated have come to
be appreciated. As such, insolubility issues of the past have provoked a paradigm change,
which now offers novel solutions for innovative drugs of the future.

SOLID LIPID NANOPARTICLES

Sub-micron-sized spherical lipid
carriers of lipid-soluble drug
molecules. They are typically
formed by heating an aqueous
lipid mixture above the melting
point of the lipid, adding drug,
homogenizing, then cooling to
freeze the drug within the the
solid lipid spheres.
HIGH-THROUGHPUT
SCREENING
An automated instrumental
process for detecting the
binding of tens of thousands of
drug candidates to an isolated
protein receptor target that
mediates a disease process,
thereby identifying worthwhile
leads for development.
Senior Director, Strategic
Technical Development,
Baxter Healthcare
Corporation, Baxter
Technology Park, Round
Lake, Illinois 60073, USA.
e-mail: barrett_rabinow@
baxter.com
doi:10.1038/nrd1494
Nanosuspensions of drugs are sub-micron colloidal
dispersions of pure particles of drug, which are stabilized
by surfactants1. For the purpose of this review, they are
distinguished from nanoparticles, which are poly-
meric colloidal carriers of drugs2, and from SOLID LIPID
3
NANOPARTICLES , which are lipidic carriers of drugs.
Activity in this area has exploded recently because of the
need for solubility enhancement.
This has become a problem during the past ten years
because of the new paradigm in HIGH-THROUGHPUT
SCREENING of new drug candidates, which emphasizes the
importance of good fit with target-receptor geometry.
Candidates emerging from these screens have high
molecular mass and high LOG P, both of which are factors
that contribute to insolubility4.
When can nanosuspensions help?
Nanosuspensions can be used for compounds that are
WATER INSOLUBLE but which are soluble in oil (high log P),
although other lipidic systems, such as liposomes and
emulsions, can be used to formulate these compounds
as well. Nanosuspensions are the preferred solution,
however, when no other approach will work. In contrast
with lipidic systems, nanosuspensions can be used to
successfully formulate compounds that are insoluble in
both water and oils. This can be the case when the crystal
energy is high, indicated by a high melting point, which
reduces the tendency of the crystal to dissolve, regard-
less of the solvent5. Nanosuspensions overcome delivery
issues for these compounds by obviating the need to
dissolve them, and by maintaining the drug in a pre-
ferred crystalline state of size sufficiently small for
pharmaceutical acceptability.
In addition, utilization of the dense, solid state confers
an additional advantage of higher mass per volume
loading. This is crucial when high dosing is required.
Such cases often fail with approaches involving molecular
complexation (for example, CYCLODEXTRINS) because of
the high molar ratio of complexing excipient that must
be used. A related benefit of high loading is reduced
administration volume, which is crucial for low-volume
intramuscular and ophthalmic applications6,7.
Conventional approaches often attempt to solubilize
insoluble drugs with the use of excessive amounts of co-
solvents, but this poses toxicity problems. Subsequently,
a related issue arises during drug development when
the safety of the agent must be studied in animals at
many multiples of its intended dose in man. The need
to administer very large doses of drugs must then be
accomplished without the interference of toxic side
effects caused by co-solvents8,9. Conducting these
studies might now be possible by using nanosuspen-
sions that dispense with solvents. As a result of these
advantages, drug candidates once abandoned could
now be rescued (FIG. 1).
The above-described requirements have driven the
development of nanosuspension technology, which has
subsequently revealed secondary benefits that are now
beginning to be realized (TABLE 1). For example, the par-
ticulate nature of the dosage form might offer alternative

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REVIEWS

Increasing drug polarity

Cosolvents
pH adjustment Micellar dispersions
Co-solvents Inclusion complexes Nanosuspensions
Salt formation Emulsions
Other lipid systems

Yes No
Suitable
molecular shape?

Dose?
Yes
High Low Low

Melting point?
High

Low High
Water soluble? Log P?
No

Can a salt be made?

Yes No

Start

Figure 1 | Decision tree for selection of formulation approach. The easiest applicable approaches are utilized. If a salt can be
made, or solubility increased by simple pH adjustment, these are done preferentially. If the drug is not particularly insoluble, co-solvents
are tried next. If there is adequate solubility in lipidic systems, then micelles, emulsions and so on are tried. Inclusion complexes, as
with cyclodextrins, can be considered. For the most intractable compounds — those with high Log P, high melting point and high
dose — nanosuspensions are used.

LOG P
Log of the octanol–water
partition coefficient, which is a
measure of a drug’s lipophilicity.
Defined as the ratio of
un-ionized drug distributed
between the octanol and water
phases at equilibrium. Higher
values imply greater lipophilicity.
WATER INSOLUBLE
Less than 0.1 mg per ml
solubility in water.
CYCLODEXTRINS
5–8mer of cyclic linked amylose
or glucan molecules that forms
a hydrophobic interior to
accommodate an insoluble
compound, and a hydrophilic
exterior to solubilize in water.
BIOAVAILABILITY
A measure of the rate and
extent of drug absorption from
an administered dose, expressed
as a ratio to an intravenously
administered dose.
pharmacokinetic profiles in intravenous delivery that
can offer less toxic and more efficacious regimens. By
decreasing the particle size of the solid form of the drug,
the dissolution rate is increased, thereby addressing a
number of issues related to poor oral BIOAVAILABILITY. The
solid state of the drug offers solutions to issues of chem-
ical stability, and the small size confers increased physical
stability with respect to sedimentation. Although manu-
facturing complexity is increased, it is offset by utilizing
unit operations that are already being deployed in other
commercialized products. To best achieve the end bene-
fits of nanosuspension dosage forms, a firm understand-
ing of formulation theory as well as manufacturing
practices is essential.
Formulation theory
Nanoparticles can be formed by building particles up
from the molecular state, as in precipitation, or by
breaking larger micron-sized particles down, as in
milling. In either case, a new surface area, ∆A, is formed,
which necessitates a free-energy (∆G) cost as defined by
∆G = γs/l •∆A, in which γs/l is the interfacial tension. This
arises because water molecules incur fewer attractive
forces with other water molecules when located at a free
surface. The system prefers to reduce this increase in
surface area by either dissolving incipient crystalline
nuclei, in the case of precipitation, or by agglomerating
small particles, regardless of their formation mecha-
nism. This tendency is resisted by the formulator
through the addition of surface-active agents, which
reduce the γs/l and therefore the free energy of the system
(FIG. 2). These agents confer immediate protection and
are more effective when present at the time of creation
of the new, fresh surface than if added afterwards.
By virtue of their complementary properties, surfac-
tants of two classes are utilized: charged or ionic sur-
factants, which effect an electrostatic repulsion among
the particles; and non-ionic polymers, which confer a
steric repulsion — that is, they resist compression. The
mechanics of these surfactant-stabilized suspensions
can be considered from the perspective of potential
energy (BOX 1).
If the particles approach each other too closely, they
will agglomerate. This must be prevented to ensure a
stable system. Energetically, this requires the placement
of a sufficiently high energy barrier at relatively long

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Table 1 | Benefits of nanosuspensions

Physicochemical characteristic
Increased drug amount in dosage form Intravenous: reduced toxicity, increased efficacy
without harsh vehicles (extreme pH,
co-solvents)
Reduced particle size: increased drug
dissolution rate
Solid state: increased drug loading
Solid state: increased stability
Particulate dosage form
Potential benefits
Oral: increased rate and extent of absorption, increased bioavailability of drug:
area under plasma versus time curve, onset time, peak drug level, reduced
variability, reduced fed/fasted effects. Pulmonary: increased delivery to deep lung
Reduced administration volumes; essential for intramuscular, subcutaneous,
ophthalmic use
Increased resistance to hydrolysis and oxidation, increased physical stability
to settling
Intravenous: potential for intravenous sustained release via monocyte phagocytic
system targeting, reduced toxicity, increased efficacy. Oral: potential for reduced
first-pass hepatic metabolism

separation distances, to prevent the particles coming Manufacturing in practice

ARRHENIUS EXPRESSION
A mathematical equation
that relates the rate of a
chemical reaction, k, to change
in temperature, T:
k = Ae–E /RT. E is the activation
energy or barrier for the crucial
step in the reaction, and R is the
gas constant.
too close together. Therefore, a non-ionic polymeric
surfactant is also used that coats the surface with a
hydrophobic chain, and permits a hydrophilic tail to
project into the water. Compression of the polymeric
coating, as by the approach of a similarly coated particle,
causes loss of entropy and is therefore unfavourable.
This provides the necessary repulsive barrier between
two neighbouring particles. The polymeric coating
performs a dual role: inhibiting crystal growth and
reducing particle size10.
Both electrostatic and steric mechanisms are enabled
by combining polymers and ionic surfactants, which
therefore complement each other. Entropic steric
interactions are inherently more sensitive to tempera-
ture fluctuations than is electrostatic repulsion.
Therefore, temperature cycling could disrupt a sus-
pension stabilized only by polymer. To prevent this,
ionic surfactants are used as well. There is a synergy
between the two, because adding a neutral polymer
to a surface stabilized with an ionic surfactant permits
greater coverage by the ionic surfactant. This occurs
because self-repulsion of the charged surfactant
molecules is minimized, which therefore permits
closer packing.
It is found for a variety of colloidal systems that the
attraction energy does not vary significantly, but that
the repulsive energy fluctuates with surface potential.
Therefore, regardless of the nature of the bulk parti-
cles, colloidal stability is determined primarily by the
choice of surfactants, which affects the repulsive
potentials. This does not mean that there is no effect
of drug on the stability of suspensions; the drug
affects the nature of the surface to which the surfactant
must bind.
The design of a stable formulation for nanosuspen-
sions can differ from that used for suspensions of larger
particle size. The latter often strives for weak flocculation,
which corresponds to the secondary minimum, Vs, of
FIG. 3, and yields readily dispersible particles after agita-
tion. This is designed to prevent slow settling, which
promotes tight packing and consequent caking of the
suspensions, which are then no longer dispersible11. (For
a description of settling rate see BOX 2.)
For the manufacturing of nanosuspensions, precipitation
conditions are chosen so as to minimize particle size.
There are two phases involved: the initial creation of
crystal nuclei, and their subsequent growth. Formation
of a stable suspension with the smallest particle size
requires a high nucleation rate but low growth rate. Both
process rates are dependent on temperature, described
by an ARRHENIUS type of expression, as well as on super-
saturation. The optimum temperature for nucleation
might lie below that for growth, which permits selective
optimization for nucleation12. High-supersaturation
conditions are chosen for rapid nucleation by metering
small amounts of a water-miscible organic solution of
the drug to the non-solvent water, under rapid mixing.
This rapid dilution of solvent results in high-super-
saturation conditions, which causes spontaneous
nucleation and a subsequent reduction of the super-
saturation condition in the immediate vicinity of the
nucleating crystals; this reduces growth rates. This is
the opposite of the conditions typically chosen for
industrial crystallization. In the industrial situation, slow
growth is chosen to generate relatively larger, purer
crystals, and the number of very fine particles is delib-
erately reduced because of their interference with subse-
quent downstream processing. Slow growth is also more
likely to directly result in the often preferred, thermo-
dynamically more stable, crystal form. By contrast amor-
phous or less stable forms are initially favoured by rapid
crystallization, but can subsequently convert to more
stable forms, according to Ostwald’s ‘law of stages’13. This
instability poses an obstacle unless secondary processing
is utilized to modify the crystal form (see below).
The rapid-nucleating conditions have two additional
consequences, which can be exploited to further reduce
particle size. First, at high-supersaturation levels, the
crystal habit, or external appearance (as opposed to
the internal, periodic crystal lattice structure), is changed
to a needle-like or dendritic morphology. These crys-
tals are easily broken, forming additional nuclei,
which catalyses the nucleation process at the expense
of growth12. Second, in comparison with crystals
grown more slowly, rapidly grown crystals tend to be
more imperfect, and often incorporate impurities and

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Attrition —
for example,
milling Dissolution
Agglomeration Nucleation and
crystal growth
a b
Water molecule Water–surfactant interaction
Figure 2 | Creation and stabilization of nanoparticles, from the perspective of surface
energetics. a | In the case of unfavourable energetics, hydrophobic crystal surface directly
contacting water molecules leads to crystal agglomeration, because water molecules are
energetically driven to leave the surface, as shown by the arrows. b | A surfactant-stabilized
crystal surface reduces interfacial tension by allowing attractive water–surfactant interactions.
The crystal surface is stabilized and shows reduced tendency to agglomerate.
dislocations. This occurs because of the insufficient time
afforded for accurate incorporation of solute in to the
growing crystal lattice. This effect is more pronounced
for flexible molecules that have many degrees of freedom.
Both the dendritic morphology, as well as the crystal
defects, can be utilized in subsequent processing by
homogenization to reduce particle size further.
Homogenization. Homogenization14,15 involves the
forcing of a suspension under pressure through a
valve16,17 that has a narrow aperture18,19. Bernoulli’s law
requires that the high velocity of the suspension that
results from flow past the constriction is compensated by
a reduction in static pressure. This, in turn, causes bubbles
of water vapour to form, which then collapse as they exit
the valve. These cause cavitation-induced shock waves,
which crack the particles. Turbulent flow and shear have
also been investigated as mechanisms for reducing par-
ticle size through homogenization20–22. Crystals are sus-
ceptible to such breakage to varying degrees. Pure crystals
have a theoretical tensile strength that is 100–1,000 times
stronger than what is observed in practice. However, the
inevitable crystal defects — dislocations and impurities
— weaken mechanical behaviour by migrating and accu-
mulating at crystal-grain boundaries, which furnishes a
nidus of weakness23–25.
Box 1 | Potential energy of surfactant-stabilized suspensions
The repulsive energy of two similarly charged particles is given by the equation
VR= (εaψ0/κ2)ln[1+exp(-κHo)], in which a is the particle radius, Ho is the distance of
separation between the two particles, ε is the dielectric constant of the medium, ψ0 is the
electrostatic surface potential, and κ is related to the thickness of the diffuse electric
double layer114. The net repulsive energy decreases with separation of the particles (FIG. 3).
At shorter distances of separation, there is an attractive force between the two particles
due to van der Waals forces. The superposition of both these forces results in an attractive
potential well, provided the particles can overcome an energy barrier.
788 | SEPTEMBER 2004 | VOLUME 3
Homogenization is advantageously utilized to exploit
three consequences of rapid precipitation to either
further reduce particle size or to resolve other potential
difficulties26. First, the crystal defects induced by rapid
precipitation render the crystal more susceptible to
rupture by the subsequent mechanical shock of homo-
genization. Second, fracture mechanics predict that the
dendritic morphology itself is more susceptible to
breakage because of the narrow dimension induced,
which must bear the full applied force (FIG. 4). Third, the
mechanical energy supplied by the homogenizer enables
initially formed, unstable amorphous particles that
result from rapid precipitation to undergo subsequent
crystallization to a stable state.
Wet milling. Another manufacturing technique for
preparing nanosuspensions is wet milling, in which the
active agent, in the presence of surface stabilizer(s), is
comminuted by milling media. Particle size here is
determined by stress intensity and the number of con-
tact points. The stress intensity is a function of the
kinetic energy of the grinding beads, and the number of
contact points can be increased by utilizing smaller
grinding media. A drive shaft, attached to rotating
disks, provides the energy to a charge of crosslinked
polystyrene beads to comminute the drug crystals by a
compression–shear action27.
Other techniques. Yet another technique involves the
spraying of a drug solution in a volatile organic solvent
into a heated aqueous solution. Rapid solvent evapo-
ration produces drug precipitation in the presence of
surfactants28.
Lyophilization of a nanosuspension can be performed
to overcome either physical or chemical incompatibility,
permitting recovery of the original particle size after
reconstitution29. Sterilization can be performed by
aseptic processing of previously sterilized components30,
membrane filtration for particles sufficiently small31,32,
or, for drugs that will withstand it, steam sterilization33
or γ-irradiation.
The manufacturing process is built around the unit
operations of particle-size reduction and sterilization.
A schematic for an aseptic microprecipitation/homo-
genization process is shown in FIG. 5. The process can be
varied to permit the use of previously sterilized raw
material, sterile addition of excipients before filling, and
lyophilization. The unit operations of sterile crystal-
lization, aseptic dispersion of previously sterilized drug
and aseptic milling are all used in currently manufac-
tured microsuspensions, such as testosterone, insulin,
dexamethasone acetate and penicillin G34.
Characterization techniques
Tests for the quality of nanosuspensions are selected on
the basis of the performance required. For pharmaceuti-
cal use, a suspension must have a minimal tendency to
agglomerate, which could lead to the formation of a
hard cake. A suspension with slow sedimentation rate is
acceptable, perhaps even preferable, provided the prod-
uct is re-suspendable and homogeneous. In addition,
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VR
VT
VR
VM
Potential energy
Distance
H0
0
VS
VA
VA
VD
Figure 3 | Potential energy curve for approach of two nanoparticles. The total potential
energy curve VT is a superposition of an attractive curve VA and a repulsive curve VR. As two
particles approach, they can overcome the energy barrier VM, leading to attractive aggregation of
the closely packed particles. To prevent this for microparticulate suspensions, one can formulate
as a weak floc, making use of the secondary minimum VS114.
the product must be non-toxic and non-irritating.
A suspension used for injectables must also be sterile,
non-pyrogenic, syringeable, injectable, isotonic and non-
haemolytic35.
Particle size growth is largely responsible for agglom-
eration, as discussed earlier. A variety of techniques are
used to measure particle-size distribution, because of
their different operating principles and features. For
example, laser light diffraction can be used because it is
POLYMORPH fast and suitable for screening large numbers of samples,
One of several distinct crystal acquiring data in the useful range of 0.02–2,000 µm36.
packing arrangements,
However, because it is an ensemble37 rather than a single-
potentially having a different
energy of formation and particle-counting technique, it should be calibrated
melting point than other against a more accurate method. When the particles
polymorphic forms of the same are smaller than the wavelength of visible light, optical
molecule. Its formation might microscopy cannot be used as a reference method. But
be favored in preference to other
polymorphs by manipulation of
in this range, field emission low-voltage scanning elec-
temperature, solvent and degree tron microscopy can be used to image individual par-
of supersaturation. ticles. However, analysing a sufficiently large statistical
Box 2 | Settling rate
Settling rate is given by the Stokes equation (D2/18η)(dp – dm)g, in which D is the particle
diameter, η is the viscosity of the medium, dp and dm are the densities of the particles and
the medium respectively and g is the gravitational constant. The critical size for settling can
be calculated from the criterion of Overbeek114, which states that according to Stokes law
colloidal particles that settle at a rate of only 1 mm in 24 hours, will never settle in actual
practice because of Brownian motion. For a density of particles of 1.15 and η of 1, the
critical particle diameter is calculated at 300 nm. This can alter the stabilization strategy for
suspensions that have particle size constrained to this limit.
NATURE REVIEWS | DRUG DISCOVERY
REVIEWS
number of particles to achieve reasonable levels of
precision becomes tedious. Photon-correlation spec-
troscopy is rapid, but only covers the range of 0.02–3
µm38. Light-obscuration counters have a lower sensitivity
cut-off of 0.5 µm36. In addition to characterizing the
mean particle size and particle-size distribution, focus
should also be directed to characterizing the very high-
end particle size, especially for injectables. Although
not intended for this purpose, the United States
Pharmacopeia microscopic particle matter test, with
limits on number of particles greater than 10 µm and 25
µm, is serviceable39. Finally, visualization of particle
shape by atomic-force microscopy has been reported40.
In the course of the development of a nanosuspen-
sion formulation, one first screens various surfactant
packages to achieve the desired particle size, with a
narrow distribution in size. Once achieved, accelerated
stability tests are performed, which challenges the
system both thermally and mechanically. The rapid
assessment of sedimentation potential by near-infrared41
is possible. POLYMORPH stability to processing is assessed
by differential scanning calorimetry42–44 and X-ray
diffraction. On-line monitoring of particle size by
near-infrared45 is a possibility.
The bioavailability of a nanosuspension, by any
route of administration, ultimately depends on the
dissolution of the drug. In vitro dissolution testing in
a bio-relevant medium46,47 furnishes guidance as to
the potential pharmacokinetics to be expected in vivo,
as explained in the applications section. Dissolution
rates can be affected by pH and the nature of the poly-
morph, which can therefore affect pharmacokinetics.
By analogy to recommended practice for liposomal
dosage forms in clinical trials48, the drug levels in
blood should be separated into a free component (mol-
ecular and protein bound) and nanosuspension-bound
fraction (BOX 3).
Applications
Nanosuspensions are used to advantage in diverse dosage
forms. In some cases, their small size and increased
surface area leads to an increased dissolution rate and
increased bioavailability. In other cases, their particulate
nature dictates targeting of the MONOCYTE PHAGOCYTIC
SYSTEM (MPS), with unusual pharmacokinetic conse-
quences. Nanosuspension drugs that are marketed or
currently in clinical trials are listed in TABLE 2.
Oral. In general, oral suspensions are selected because of
the superior taste-masking of the particulate form, or
difficulty experienced by the very young and old in
swallowing tablets49, or to overcome solubility problems.
Oral nanosuspensions have been specifically used to
increase the rate and extent of the absorption of drugs50
due to their solubility limitations. In comparison with a
conventional 10-µm danazol suspension, a 169-nm
nanoparticulate dispersion showed higher CMAX (3.01
versus 0.20 µg per ml) and AREA UNDER THE CURVE (AUC)
(16.5 versus 1.0 µg•h per ml) in a pharmacokinetic study
in dogs. The bioavailability of the nanosuspension was
equivalent to that of a cyclodextrin solution formulation,
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MONOCYTE PHAGOCYTIC
SYSTEM (MPS)
The system of cells deployed
throughout the body tissues,
derived from monocyte
precursor cells, which
functionally police interstitial
fluid and blood for unwanted
particulate contamination,
removing them by phagocytosis.
CMAX
Highest drug concentration
in a plot of the plasma drug
concentration versus time.
AREA UNDER THE CURVE
(AUC). Area under the curve
of a plot of plasma drug
concentration versus time,
measured after administration
of a drug.
FED/FASTED EFFECTS
A difference in the absorption
of a drug depending on the fed
or fasted state of the test animal.
790 | SEPTEMBER 2004 | VOLUME 3
b After precipitation
a Raw material Before homogenization c After homogenization
20 microns 20 microns 20 microns
Figure 4 | Engineering breakable crystals with a combination of microprecipitation and homogenization. Crystal
morphology of raw drug material is modified to facilitate breakage into smaller nanoparticles. a | Crystals of starting raw material are
too large and hard to run efficiently through a homogenizer. b | The raw material is solubilized, filter sterilized and precipitated, so as
to yield crystals of needle-like morphology, which are easily broken during homogenization c | Homogenization yields nanoparticles
suitable for parenteral injections.
indicating that the dissolution-rate-limited bioavailabilty the particles are shed from the mucus55,56. Perturbed
observed with the 10-µm suspension had been over- absorption resulting from an initial coating of salivary
come. The absolute bioavailability was 82.3 ± 10.1% of proteins on nanoparticles might be prevented by enteric
an intravenous control injection51. coating57. Targeting to macrophages in inflamed colon
Evidence for enhanced onset of action, reduction mucosa might also be possible58.
of FED/FASTED ratio (G. Liversidge, personal communi-
cation), and enhanced dose proportionality have also
been found. There is even evidence for decreased gas-
Solution of drug in
tric irritancy by nanosizing naproxen52. In this case, Aqueous buffer solution
water-miscible solvent with
with optional surfactant
reducing the particle size from 20–30 µm to 270 nm optional surfactant
led to faster absorption (tmax = 23.7 versus 33.5 min).
The implied decrease in gastric residence time, with
associated local high and prolonged concentration of
naproxen, was presumed to be responsible for the Sterilizing filter Sterilizing filter
reduced gastric irritancy scores. These effects are med-
iated primarily by an increase in dissolution rate sec-
ondary to increased surface area, as elaborated in the
Noyes–Whitney equation dC/dt = [DA/hV][Cs–C], in
which dC/dt is the dissolution rate of a drug formula- Suspension of amorphous or semi-crystalline particles
tion, D is the diffusion coefficient, h is the thickness of
the diffusion layer at the solid–liquid interface, A is the
surface area of drug exposed to the dissolution media,
V is the volume of the dissolution media, Cs is the Homogenize
concentration of a saturated solution of the solute in
the dissolution medium at the experimental tempera-
ture, and C is the concentration of drug in solution
at time t53. Stable crystalline product
Although reductions in particle size increase uptake
by both dissolution and particulate pathways (see
Remove solvent
below), increased self-agglomeration of small particles
can, paradoxically, lead to reduced uptake, which must
therefore be anticipated. Fill/finish
In addition, increased surface area and decreased
particle size can lead to increased muco-adhesion, which Figure 5 | Schematic of aseptic microprecipitation/
can increase gastrointestinal transit time and lead to homogenization process. Although the final dosage form is
particulate in nature, the drug can be filter sterilized by initially
increased bioavailability. However, this result is compli-
dissolving it in a water-miscible organic solvent, and passing
cated by the effect of both surface charge54 and surfactant the resulting solution through a sterilizing filter. All subsequent
type of the nanoparticles. These parameters determine steps are conducted in an aseptic environment (previously
the particles’ adsorption to, and penetration through, the sterilized and maintained as such). Following a solvent removal
mucus layer to reach the underlying epithelial cells before step, the aqueous suspension is filled into vials.
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OSTWALD RIPENING
The tendency for a particle
dispersion to grow in diameter
over time, by a process in which
the smaller particles dissolve
preferentially, because of their
higher solubility, with
subsequent crystallization onto
larger particles, making them
even larger.
PHAGOLYSOSOME
A cytoplasmic vesicular
compartment consisting of the
union of a phagosome with a
lysosome, containing enzymes
and acid environment designed
to digest microbial particles.
t1/2
Half-life of plasma drug
concentration, a measure of the
duration of action of a drug.
NATURE REVIEWS | DRUG DISCOVERY
By reducing the size of particles to the sub-micron
level, the uptake of intact gastrointestinal polymeric
particles has been shown to occur preclinically59,60, by
mechanisms involving M-cells in Peyer’s patches of the
gastrointestinal lymphoid tissue61. This uptake pathway
communicates with the mesenteric lymph ducts, and
empties via the thoracic duct into the systemic blood
circulation. This approach therefore provides a route
for avoiding first-pass metabolism, as well as for target-
ing sanctuaries of lymphatic-mediated diseases62. The
low drug uptake by this pathway might be enhanced, as
shown in cell-based studies, by coating drug particles
with agents, such as vitamin B1263, that dock into trans-
porter receptors on the intestinal epithelium. The use
of surfactants has also been found to have an impact on
decoupling the intestinal P-glycoprotein drug-efflux
pump as well as interfering with lipidic chylomicron
transport systems.
Injectable. Injection of poorly water-soluble drugs is
often approached by formulating drugs with excessive
amounts of co-solvents64, which provoke anaphylactoid
reactions65,66, pain on injection67 and precipitation of
drug following dilution of the vehicle in the blood68,69.
Lipids might work for drugs that are lipid-soluble70,
which often is not the case for drugs having high crystal
energies71. To establish a more comprehensive approach,
therefore, the formulation of injectable drugs as
nanosuspensions has emerged. The approach is gener-
ally applicable, provided that the aqueous solubility is
below several hundred µg per ml, to prevent OSTWALD
RIPENING. Successful formulation has been reported as
applied to antineoplastic agents72, anaesthetic agents73,
antifungals and antibacterials29, as well as for agents for
malignant hyperthermia74 and cancer pain75.
Several pharmacokinetic profiles can result following
the injection of nanosuspensions. If the particles dissolve
in the blood readily, both the pharmacokinetics, and
therefore tissue distribution, will be equivalent to those
for the solution formulations76,77, affording a relatively
fast onset of action78. Alternatively, depot delivery via
subcutaneous, intramuscular or intradermal routes
offers prolonged drug release, because of the ability to
load more drug safely into a small injectable volume79.
The greater loading capacity (up to 30%) distinguishes
nanosuspensions from polymeric nanoparticulate
vehicles. The small size of the particles results in sig-
nificantly faster dissolution than microsuspensions,
provided that post-injection aggregation does not
occur. Because drug dissolution is often the rate-limiting
step in systemic drug uptake from a depot80,81, nano-
suspensions often result in higher peak plasma levels as
well. But because dissolution is not instantaneous, as it
would be for a solution dosage form, there is less toxicity
associated with nanosuspensions, enabling high loading
with safety73.
If intravenously administered particles do not dissolve
immediately, they are subject to uptake by macrophages
of the MPS82, primarily the Kupffer cells in the liver and
those in the spleen. As the phagocytized drug particles
encounter the reduced pH of the PHAGOLYSOSOMES83, their
REVIEWS
Box 3 | Characterization tests
Chemical
• Active ingredient
• Degradation products
• Moisture (for lyophilized and solid dosage forms)
• Preservatives
• pH
Physical
• Particle-size distribution
• Particle-size distribution in response to accelerated
ageing and shipping (freeze/thaw, mechanical agitation,
centrifugation)
• Drainability (from sides of container)
• Syringeability, injectability
• Re-suspendability
• Dissolution in water or biorelevant medium
• Compatibility after admixture
• Zeta potential (electrostatic self-repulsion of particles)
Biological
• Sterility
• Pyrogenicity
• In vivo pharmacokinetics
solubility–pH profile might permit dissolution of the
compound. Their lipophilic character might permit
passage through the phagolysosomal membrane, and
as a result they could leave the cellular vesicle, enter
the cytoplasm, and then exit the cell by diffusing
down the drug concentration gradient as shown in
FIG. 6. This intravenous depot effect will result in a
pharmacokinetic profile with significantly reduced
Cmax, but quite prolonged t1/2. This can be very advan-
tageous for certain drug classes, for which toxicity is
mediated by peak plasma values, but for which efficacy
is driven by AUC, as in the case of triazole antifungals84.
Formulation of the antifungal agent itraconazole as a
nanosuspension resulted in significantly less toxicity
after intravenous injection than the commercially
available cyclodextrin-solubilized drug itraconazole
(Sporanox; Janssen Pharmaceutica). This permitted a
much higher drug dose to be injected into rats and
dogs, with a consequent increase in efficacy, as shown
by mortality and colony counts in Candida albicans-
challenged immunosuppressed rat models. The
greater tissue drug loading that resulted enabled the
successful treatment of fungal strains that are normally
resistant to itraconazole in animal models treated
with nanosuspension85.
The pharmacokinetic parameters of a nanocrystal
suspension of itraconazole, as determined in a clinical
trial86, can be compared with those of the commercial
Sporanox solution, as listed in the Physician’s Desk
Reference 87 and other literature. The nanocrystal suspen-
sion occupied a larger volume of distribution (1,677 ±
827 l versus 796 ± 185 l) and was cleared more slowly
(3.35 ± 1.8 l per h versus 22.9 ± 5.7 l per h) to give a
VOLUME 3 | SEPTEMBER 2004 | 7 9 1
REVIEWS

Table 2 | Solid-particulate-nanosuspension-based formulations in development and in the market

Drug Indication Drug delivery company Pharma company Route Status
Paclitaxel Anticancer American BioScience American Pharmaceutical Intravenous Phase III
Partners
Undisclosed Anti-infective Baxter NANOEDGE Undisclosed Oral/ Preclinical
multiple intravenous to Phase II
Undisclosed Anticancer Baxter NANOEDGE Undisclosed Intravenous/ Preclinical
oral to Phase I
Rapamune Immuno- Elan Nanosystems Wyeth Oral Marketed
suppressant
Emend Anti-emetic Elan Nanosystems Merck Oral Marketed
Cytokine Crohn’s Elan Nanosystems Cytokine Oral Phase II
inhibitor disease PharmaSciences
Diagnostic Imaging agent Elan Nanosystems Photogen Intravenous Phase I/II
Agent
Thymectacin Anticancer Elan Nanosystems NewBiotics./Ilex Oncology Intravenous Phase I/II
Fenofibrate Lipid lowering SkyePharma Undisclosed Oral Phase I
Busulfan Anticancer SkyePharma Supergen Intrathecal Phase I
Budesonide Asthma Elan Nanosystems Sheffield Pharmaceuticals Pulmonary Phase I
Silver Eczema, NUCRYST Self-developed Topical Phase I
atopic
dermatitis
Calcium Mucosal BioSante Self-developed Oral
phosphate vaccine adjuvant
for herpes Phase I
Insulin Diabetes BioSante Self-developed Oral Phase I

PHOSPHOLIPID-PEG
A molecule consisting of
phospholipid covalently
bonded to a polyethylene glycol
polymer, used as a reagent to
coat a drug or drug delivery
vehicle with a polyethylene
glycol (PEG) surface, thereby
conferring longer blood
circulation time.
longer half-life (346 ± 225 h terminal versus 35.4 ± 29.4
h mean and 30 h terminal88) and larger area under the
plasma concentration curve for the first 24 hours,
AUC24, (51,558 ± 10,635 µg•h per l versus 30,605 ±
8,961 µg•h per l). This behaviour is consistent with
MPS depot behaviour, resulting in prolonged delivery
for the nanosuspension. The nanocrystal suspension
was well tolerated.
A macrophage uptake mechanism also explained
the behaviour of a clofazimine nanosuspension. After
intravenous administration, both a liposomal and a
nanosuspension formulation of this drug had similar
drug distribution to the liver, spleen and lung, and
achieved effective control of a macrophage-inhabiting
mycobacterium29.
The particles will remain in the macrophages if they
are too insoluble, or if they cannot be metabolized to
soluble compounds. To remedy this problem, modifica-
tion of molecules has been used to increase clearance
following phagocytic sequestration. In this case,
nanoparticulate X-ray contrast agents were injected sub-
cutaneously, such that their subsequent cellular uptake
and delivery to lymph nodes89 resulted in opacification
of these organs. Modification of the molecules enabled
their subsequent clearance and removal.
Modification of the surface of slowly dissolving drug
crystals with PHOSPHOLIPID-POLYETHYLENEGYLCOL (PEG)
chains90 could delay protein adsorption and opsonization
of the particles, thereby reducing macrophage uptake.
This would increase circulation time, and give the particles
the opportunity to find, and leak out of, discontinuities of
vasculature, such as occurs in neoplasms, infections and
inflammation. In effect, this would result in passive target-
ing by the enhanced permeation and retention effect for
tumours, and sites of infection and inflammation91,92.
Pulmonary. To target the deep lung, respirable aerosols
should have mean aerodynamic diameters of 1–5 µm93.
In conventional solid-in-liquid dispersions, the solid drug
has a particle size that is comparable to that of the aerosol.
There is therefore statistical inhomogeneity in the par-
titioning of drug particles among the carrier droplets94.
Nanosuspensions would ameliorate this by increasing the
number of particles per droplet95. In fact, aerosol cascade
impactor studies have shown significantly higher res-
pirable fractions and lower unwanted systemic uptake
via throat deposition for nanosuspensions compared
with micronized formulations96. Respirable dose was
increased from 227 µg to 421 µg by decreasing particle
size from 4.4 µm to 0.73 µm, prepared by homo-
genization97. The use of already approved surfactants
can be used for drug nanosuspensions98, to avoid safety
problems posed by polymeric nanoparticulates99.
In a Phase I clinical trial, nebulized nanocrystal
(75–300 nm) budesonide suspension had double the
Cmax, and almost half the tmax, of larger, 4,400-nm sized
Pulmicort Respules. The total amount absorbed, as
indicated by AUC, was comparable. The faster absorp-
tion, which resulted in higher peak plasma levels,
could have been attributable either to more rapid dis-
solution or faster entry to the blood via access to the
additional vasculature of the peripheral lung100.
Nanosuspension targeting to the alveolar macro-
phages is an attractive option for diseases mediated by

792 | SEPTEMBER 2004 | VOLUME 3 www.nature.com/reviews/drugdisc

 REVIEWS

these cells, both to elevate drug concentration within the
target cell, and to decrease concentration systemically,
thereby reducing potential toxicity. With consequently
prolonged regional drug levels, the minimization of
dose frequency might also be anticipated. This could
be useful in tuberculosis, in which the mycobacteria
reside in macrophages. Efficacy would occur both by
enhanced killing of the intracellular parasites as well as
by utilizing the trafficking of alveolar macrophages to
secondary lymphoid organs — mimicking the migration
path of the mycobacteria — as a kind of Trojan
Horse101,102. Enhanced uptake of 0.5-µm versus 3-µm
sized particles by non-macrophage respiratory epithelium
cell lines has also been demonstrated103.
For the efficient delivery of particles to the lungs,
clearance mechanisms must be overcome. These consist
of the muco-ciliary escalator in the upper airways, and
alveolar macrophage uptake deep in the lung. As shown

Opsonins
complement,
immunoglobulins

Drug/ Opsonized
nanoparticle nanoparticle
Pseudopod extension Pseudopodia fusion
occurs as surface receptors dock leads to topological
onto opsonizing ligands internalization

Receptor-
mediated
recognition

Crosslinking of cell-
surface receptors
Cytoplasmic
generates intracellular
membrane
signals, reorganizing actin-
based cytoskeleton
to extend pseudopodia Budding,
recycling
of membrane
Phagosome
Recycling compartment
pH 6

pH 5–6
Nucleus
1 µm

Enzyme 500 nm
vesicle
TEM of drug
Trans-Golgi network Lysosome crystals in phagosomes

Phagolysosome
pH 4–5.5

Ideal pH–solubility curve of

drug for phagolysosomal escape

Drug
Solubility

Cytoplasm
pH 7.4

pH 5.0 pH 7.4 Extracellular drug release

Figure 6 | Trafficking of drug nanoparticles by macrophages, monocytes and neutrophils. Nanosuspension drug particles,
opsonized by proteins, dock onto receptors on the cell surface. This initiates phagocytosis or internalization of the particle. Membrane
material is recycled back to cell surface via the recycling compartment, as enzyme vesicles from the trans-Golgi network fuse with
the phagosomes, while the pH is progressively decreased. Fusion with lysosomes lowers pH further. Depending on the pH–solubility
curve of drug, the drug can escape from depots in intracytoplasmic compartments, entering first the cytoplasm then extracellularly,
providing sustained systemic drug release83. TEM, transmission electron micrograph.

NATURE REVIEWS | DRUG DISCOVERY VOLUME 3 | SEPTEMBER 2004 | 7 9 3

REVIEWS

PHAGOCYTOSIS
Process of removal of
particulate matter, bacteria,
viruses, degraded cell debris,
and so on by any of several
mechanisms (for example,
complement activation,
opsonization or receptor
mediation) involving extension
of the cell membrane to
surround and engulf the object,
followed by severance of the
membrane leading to a distinct
vesicle within the cytoplasm of
the cell, termed the phagosome.
OMMAYA RESERVOIR
A system for the delivery of a
flowable drug formulation to the
ventricles of the brain, involving
a compartment for containment
implanted in a proximal
subcutaneous location.
PEGYLATION
Coating, either covalently or
by physical adsorption, a
polyetheylene glycol polymer
onto a drug molecule, surface
or particle, rendering it less
hospitable to the deposition
of opsonizing proteins which
mediate phagocytosis. Such
coated drugs and particles
typically manifest extended
circulation times in blood.
above, nanosuspensions can result in reduced deposition
in the former, but enhanced deposition in the latter.
The consequences of this for nanoparticles of insulin
have been noted104, for which phagolysosomal degrada-
tion of the protein drug subsequent to macrophage
uptake occurs. However, for a molecule with appro-
priate solubility-versus-pH profile, and stability in an
acid environment, macrophage uptake simply pro-
vides a mechanism for sustained release. Additionally,
modification of the surfactant coating can decrease
105
PHAGOCYTOSIS . An intriguing system incorporating
nanoparticles, held together by van der Waals forces, to
form large porous particles has been proposed to offer
the advantages of both minimal phagocytosis as well as
tenacious adsorption, thereby ensuring maximal disso-
lution106. The particles are larger than the 3-µm limit
for effective phagocytosis, therefore ensuring longer
lifetime within the lung for sustained release. But
because they are porous, their density is low, which
results in an optimal aerodynamic diameter.
Central nervous system. Nanosuspensions afford a means
of administering increased concentrations of poorly
water-soluble drugs to the brain with decreased systemic
effects107. Significant efficacy has been shown with
microparticulate busulfan in mice administered intra-
thecally108. The work has advanced to Phase I in patients
afflicted with neoplastic meningitis, administered via an
OMMAYA RESERVOIR for intraventricular delivery, and via
lumbar puncture. The drug was well tolerated and
resulted in delayed progression of disease (H. Friedman,
personal communication). Epidural injection of a 10%
butamben suspension for cancer pain was well tolerated
in dogs and humans75,109.Future work will probably also
involve less invasive routes, utilizing either passive target-
ing (via PEGYLATION, as has been done for liposomes)110
or active111–113 targeting to the brain following intra-
venous administration of nanosuspensions. In these
latter publications, it was found that use of the agent
Polysorbate 80 in the formulation led to deposition of
apolipoprotein E on the nanoparticles, which facilitated
brain uptake by receptors on the brain endothelial cells.
Concluding remarks and predictions
Nanosuspension technology initially arose to address
solubility needs of drug discovery pipelines. As experi-
ence was acquired through applications, however, more
subtle problems began to be viewed as becoming solvable
through this new technology. Altered pharmacokinetic
profiles have become appreciated insofar as they improve
safety and efficacy.
The future growth of nanosuspension technology
will mirror that of drug delivery in general, borrowing
concepts with appropriate adaptation for particulates:
applications to protein delivery will be established;
controlled stability of the amorphous phase will permit
even faster dissolution of drug nanosuspensions; ver-
satility in coating technology will be explored. In intra-
venous delivery, control over endosomal escape following
phagocytosis of drugs will become improved, and target-
ing will become more widely accepted as its value is
confirmed. Passive targeting will adapt what has been
learned from long-circulating liposomes. Blood–brain
barrier targeting will perhaps benefit from the experi-
ence with polymeric nanoparticles, and oral drug
delivery could combine nanosuspensions with receptor-
mediated particulate uptake. Such a combination,
now that dissolution problems have been largely
solved, would address the last great barrier to drug
absorption — that of permeability-limited uptake.
The solution to this problem would simultaneously
avoid first-pass hepatic metabolism as well. We predict
that an era will emerge in which soluble drugs will be
intentionally converted to insoluble complexes to take
advantage of the benefits conferred by nanosuspension
drug delivery.

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