Professional Documents
Culture Documents
com
Molecular Engineering of Biological and Chemical Systems (MEBCS), Singapore-MIT Alliance, 4 Engineering Drive 3, 117576 Singapore
b
Department of Chemical and Biomolecular Engineering, National University of Singapore, 4 Engineering Drive 4, 117576 Singapore
Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge MA 02139, United States
Received 2 April 2007; accepted 9 October 2007
Available online 13 October 2007
Abstract
Paclitaxel and poly (L-Lactic acid) (PLA) were co-precipitated to form micro and submicron particles in a manner similar to that used in the
supercritical antisolvent with enhanced mass transfer (SAS-EM) process. As compared with conventional processes, a major advantage of
supercritical CO2 as an antisolvent in the SAS-EM process is the effective removal of residual organic solvents. In this work, the organic phase was
sprayed into supercritical CO2 (for CO2, Tc = 31.1 C, Pc= 73.8 bar) from a 500 m ID capillary nozzle. Ultrasonic vibration with an amplitude of 0
to 120 m (from a 3/8 tip diameter titanium probe) was employed in the high pressure vessel during the antisolvent process to provide enhanced
mixing between the solvent and antisolvent phases. The role and effects of ultrasonication on the properties of the resulting particles were studied.
When no ultrasonication was applied, micrometer-sized particles were obtained. When ultrasonication was applied, more uniform particles in the
submicron size range were obtained. The size of the particles was found to vary with the ultrasonic vibration amplitude. Encapsulation efficiencies up
to 83.5% and controlled release of paclitaxel for more than 30 days were achieved with the particles fabricated in this study.
2007 Elsevier B.V. All rights reserved.
Keywords: Supercritical antisolvent; Paclitaxel; PLA; Biodegradable nanoparticles; Ultrasonication; Enhanced mixing
1. Introduction
Paclitaxel is a promising anticancer drug with efficacy
against a wide variety of carcinomas [14]. It works through the
inhibition of DNA synthesis by stabilizing microtubule
assembly [2]. However, its clinical application has been limited
due to its hydrophobic nature. Its current formulation requires
the use of an adjuvant, Cremophor EL, which has been
associated with several undesirable side effects [1,4]. One
method to overcome the problems brought about by Cremophor EL is to encapsulate paclitaxel in biodegradable
polymers such poly lactic acid (PLA) or poly lactic-co-glycolic
acid (PLGA). These biodegradable polymeric particles also
Corresponding author. Department of Chemical and Biomolecular Engineering, National University of Singapore, 4 Engineering Drive 4, 117576
Singapore. Tel.: +65 6516 5079; fax: +65 6779 1936.
E-mail address: chewch@nus.edu.sg (C.H. Wang).
0168-3659/$ - see front matter 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.jconrel.2007.10.002
97
Fig. 1. Simplified experimental setup for SAS and SAS-EM production of micro and submicron particles for controlled delivery of paclitaxel. Different vessel
configurations used in this study were shown in the top left hand corner. A: conventional SAS process with no ultrasonication; B: modified SAS-EM process with
spray nozzle directed away from ultrasonic vibrating surface. C: SAS-EM process with spray nozzle directed at ultrasonic vibrating surface.
98
Table 1
Paclitaxel-loaded PLA particles fabricated by modified SAS-EM method
Sample
UA
B1
B2
B3
B4
B5
B6
B7
B8
C1
10
10
0
10
20
30
40
20
10
657 223
826 385
795 314
500 181
478 142
372 86
LLS (nm)
co,
D1,0
D4,3
D3,2
theoretical
723
804
2396
921
481
465
660
846
734
816
2432
935
488
472
670
859
731
812
2420
930
485
470
666
855
0.03
0.05
0.10
0.10
0.10
0.10
0.10
Blank
0.10
co
cs
co cs
59.3 0.3
74.8 4.6
70.0 3.5
67.7 1.4
56.4 14.4
78.7 0.3
0.0198
0.0374
0.0700
0.0677
0.0564
0.0787
3.5 10 20
3.5 10 20
3.5 10 20
3 10 8
1 10 7
0.03
0.03
0.03
b0
N0
N0
UA is the ultrasonic vibration amplitude (defined as % of maximum amplitude) applied during the modified SASEM process.
co, theoretical is the theoretical loading of paclitaxel in the PLA micro- and nanoparticles.
co (mg/mg PLA) is the actual loading of paclitaxel in the PLA micro- and nanoparticles.
cs (mg/mg PLA) is the solubility of paclitaxel in PLA polymer matrix as defined in Eq. (1).
D (m2 s 1) is the diffusivity of paclitaxel in PLA polymer matrix as defined in Eq. (1).
k (s 1) is the dissolution constant of paclitaxel in PLA polymer matrix as defined in Eq. (1).
Actual loading of paclitaxel mg
a
Encapsulation efficiency, EE (%) is defined as Theoretical
loading of paclitaxel mg 100k.
99
Fig. 2. A. Close-up of the organic solution jet issuing from the 500 m ID capillary nozzle during the conventional SAS process (configuration A). The snapshot on the
left shows the breakup of the horizontal jet upon entering the supercritical CO2 phase. The snapshot on the right shows that the dense organic phase later sinks toward
the bottom of the vessel. B. Close-up of the organic solution jet issuing from the 500 m ID capillary nozzle during the modified SAS-EM process (configuration B).
The snapshot on the left shows the breakup of the horizontal jet upon entering the supercritical CO2 phase. The snapshot on the right shows the later mixing of the
organic solution phase. C. Close-up of the SAS-EM process geometry (configuration C). The snapshot on the left shows the position of the capillary nozzle used for
introducing the organic solution into supercritical CO2. The snapshot in the center shows the later mixing of the solution phase. Agglomeration of solid particles was
observed at the capillary opening. The SEM image on the right shows the typical submicron particles achieved.
100
Fig. 3. SEM pictures taken of PLA particles fabricated using different ultrasonic vibration amplitude (A) Sample B3 (no ultrasonication); (B)(D) Samples B4, B5 and
B6 (ultrasonic vibration amplitude 30, 60 and 90 m respectively).
101
Fig. 4. A. Thermogram properties for blank PLA and paclitaxel loaded PLA
particles fabricated using SAS and SASEM methods. Placebo PLA was
fabricated using the modified SASEM process without any drug loading.
B. DSC thermogram of a physical mixture of 5% paclitaxel (crystalline)/ 95%
Placebo PLA and sample B3 before and after suspension in PBS solution.
102
Fig. 5. Effect of particle size on in vitro release of paclitaxel from PLA microparticles
(B3) and submicron particles (B4, B7).
103
result is:
2
Ac
A c 2 Ac
D
k cs c
At
Ar2 r Ar
Fig. 6. (A) and (B) FESEM image of cross-section of 10% loading (B3) samples before suspension in PBS solution; (C) and (D) FESEM image of cross-section of 10%
loading (B3) samples after 2 weeks suspension in PBS solution; (E) and (F) FESEM image of cross-section of 5% loading samples after 1 week suspension in PBS
solution.
104
solubility. When the initial drug loading (co) is less than the drug
solubility, crystals are absent. When initial drug loading (co) is
higher than the drug solubility (cs), it is assumed that the excess
forms crystallites which are dispersed in the polymer matrix.
In this model, the drug-loaded particles are assumed to be
spherical and non-porous. From the FESEM pictures in Fig. 6, it
was observed that the internal structure of the PLA microparticles
was fairly compact. The surface-volume mean diameter, D32, of
each sample was obtained from SEM experiments and was used
in the fitting of the experimental release profiles to the diffusion /
dissolution model. It is assumed that the volume of the drug
crystals is negligible compared to the overall matrix volume and
that the crystallites are small and uniformly distributed throughout the polymer matrix.
A solution to Eq. (1) may be obtained in non-dimensional
form by analogy to Harland's diffusion / dissolution solution
[40]:
w
Mt
4
cs 3 kR3
l
X
Di n2 k2 Dis n2 k2 1 exp Di n2 k2 s
n1
Di n2 k2 2
2
Mt
Mt
cs
4 3 w
MT co 3 kR
co
kR2
Dt
;s 2
R
D
105
106
[36] R.T. Liggins, H.M. Burt, Paclitaxel loaded poly (L-lactic acid) (PLLA)
microspheres II. The effect of processing parameters on microsphere
morphology and drug release kinetics, Int. J. Pharm. 281 (2004) 103106.
[37] R.T. Liggins, H.M. Burt, Paclitaxel loaded poly (L-lactic acid) microspheres: properties of microspheres made with low molecular weight
polymers, Int. J. Pharm. 222 (2001) 1933.
[38] M. Polakovic, T. Gorner, R. Gref, E. Dellacherie, Lidocaine loaded
biodegradable nanospheres II, Modeling of drug release. J. Control. Rel.
60 (1999) 169177.
[39] R.T. Kurnik, R.O. Potts, Modeling of diffusion and crystal dissolution in
controlled release system, J. Control. Rel. 45 (1997) 257264.
[40] R.S. Harland, C. Dubernet, J.-P. Benoit, N.A. Peppas, A model of
dissolution-controlled, diffusional drug release from non-swellable
polymeric microspheres, J. Control. Rel. 7 (1988) 207215.