Scale up of Atorvastatin Delayed Release Nanoparticles for Treatment of Hyperlipidemia:

Quality by Design (QbD) Approach

Gite S. M., Mirani A. G., Patravale V.B..
Department of Pharmaceutical Sciences and Technology, Institute of Chemical Technology, Matunga, Mumbai-400019

AIM & OBJECTIVE INTRODUCTION Transformation of nanoformulation from lab scale to pilot scale
Wide absorption window

• Quality by Design development of Atorvastatin calcium delayed

release nanoparticles.
Atorvastatin Calcium
•
• . To evaluate the in vitro & in vivo characteristics of Atorvastatin
calcium delayed-release nanoparticles (NP)
BCS Class II Delayed Release Acid Instability QbD
Nanoparticles (ACDRNPs) Approach X
• Scale up of developed Atorvastatin calcium delayed release
nanoparticles
Low Bioavailability Aqueous phasePeristaltic Organic Evaporation Spray Drying
Pump Phase

EXPERIMENTAL
DOE by QBD DOE by QBD 32 Full Factorial Design (FFD) Preformulation Studies In- Vitro study protocol

 Quality Target Product Profile of (ACDRNPs)  Independent variables  Solubility studies in various stabilizers
More Since this is DR dosage
Cumulative
QTPP Elements Target drug Release
than 80
%
No form, CDR is moderately
critical.
Factors Low level Middle High level • USP Apparatus 4 (Sotax TM CE7,
(-) Level (+)
Dosage form Nanoparticle
Bioenhancing property of Drug: polymer ratio 1:1 1:2.5 1:5 Switzerland)
Dosage type Delayed release the drug formulations is
• Content Uniformity 200-500
highly responsible for Surfactant concentration (%) 0.25 0.5 1 • Close Loop System
Dosage strength 5/10/20/40/80 mg Particle Size
nm
Yes attaining meaningful

• Assay
Administration Oral
pharmacodynamic effects;
hence was taken up as
 32 full factorial Design • 22.6 mm cells (i.d.)
• Particle Size Alu-Alu blister
Packaging
highly critical.
Batch code
Drug: Polymer
ratio
Surfactant concentration
(%)
• Piston pump (Sotax CY7–50)
Stability of nanoparticles is
• PDI
Stability 6 months
Polydispersity
index
0.1-0.3 Yes dependent on PDI so it is ACNP1 1.00 0.25 • Dissolution Medium- 1.2 pH HCL
highly critical
ACNP2 1.00 0.50
• Zeta potential Dosage form is DR, so it is
ACNP3 1.00 1.00
followed by pH 6.8 Phosphate buffer
Encapsulation very important that EE

• DSC
Critical Quality Attributes of ACDRNPs efficiency
(EE)
60-100% Yes should be at higher side. So
it was attributed as highly
ACNP4 2.50 0.25  Polymer drug compatibility studies using FTIR and DSC 900ml at 37±0.5˚C
ACNP5 2.50 0.50
• FTIR
Quality
Attributes
Is this
a
critical parameter
ACNP6 2.50 1.00
• Flow rate- 8 ml/min
of
•
the TEM
Drug
Target
CQA
?
Justification
 Risk Assesment analysis ACNP7 5.00 0.25 • Time- 2 hr in 1.2pH HCl and then
• XRD Color
Product
Physical attributes of
ACNP8 5.00 0.50 5,15,30,45,60, 120 min
•
PhysicalIn vitro studies
Odor the formulation were
not considered as
Drug Product
CQA's
Drug:
polymer ratio
Stabilizer
conc.
Stirring
rate
Mixing
time
ACNP9 5.00 1.00

• In vivoAppear
attributes
studies
No
critical, as these are not  Experimental procedure for preparation of of Nanoparticle In- Vivo study protocol
ance directly linked to the
Fig. FTIR Spectra of AC Fig. FTIR Spectra of physical mixture
efficacy and safety. Assay and CU Low Low Low Low
Assay and content
• Male Albino rats (6–7 weeks old)
CDR Low Low Low Low
Assay and
uniformity tend to
affect safety and
weighing between 180 and 200 g
90- Particle Size High High Low Low
Content
uniformity
100%
No efficacy of
formulations, variables
• Dose: 9mg/kg
PDI Medium Medium Medium Medium
were regarded
moderately critical.
as • Route of administration: Oral
EE High High Medium Medium
• Time: 0.5, 1, 1.5, 2, 3,4, 6, 8 and 12 h
Medium risk factors are optimized on the basis of Fig.DSC thermogram of AC Fig. FTIR Spectra of physical mixture
preliminary batches

RESULT AND DISCUSSION

Analysis of Data using Design Expert Validation and scale up of nanoformulation Particle Size and zeta potential
ert® Software

Design-Expert® Software Design-Expert® Software

P Size Predicted
P Size vs . Actual Box-Cox Plot for Power Trans form s
ug:polymer
abilizer
Color points by value of
P Size:
425.3
430.00 Lambda
Current = 1
Best = 0.13
6.36
Particle size and EE of
ACDRNPs
Low C.I. = -1.16

440
262 High C.I. = 1.45
400 62
Recommend transform:
350
Encapsulation efficiency

387.50 None 5.64

395 (Lambda = 1) 61
Particle Size in nm

300
P S iz e

350
250 60
345.00 4.92
305 200 59
260 150 58
302.50 4.19
100
1.00 5.00
50 57
0.81 4.00
260.00 3.47
0 56
0.63 3.00
ACNP1 ACNP2 ACNP3
261.20 302.23 343.25 384.28 425.30 -3 -2 -1 0 1 2 3
0.44 2.00
B: Stabilizer A : Drug:polymer
0.25 1.00
Batch code
X: Actual X: Lam bda
Y : Predic ted Y : Ln(R es idualSS)

Particle Size (nm) Encapsulation efficiency (%)

Fig. 3D-Response and Model diagnostic plots of particle size

The "Pred R-Squared" of 0.9663 is in reasonable agreement with the "Adj R-Squared" of 0.9910. "Adeq Precision"
measures the signal to noise ratio. A ratio greater than is desirable. Here ratio of 41.531 indicates an adequate signal.
Software
This model can be used to navigate the design space.
ANOVA Equation: P Size =+356.60+10.66 * A-36.58 * B-44.20 * A * B-49.45 * A2+24.19 * B2
Design-Expert ® Sof tware

In vivo pharmacokinetics Studies

ymer EE Predicted
EE vs . Actual Box-Cox Plot for Power Transforms
r Color point s by value of
88 EE: 88. 00 6.50
87.5

45.6

76.5
76. 50 6.23

65 Parameters Unit Observed values

EE

65. 00 5.96

53.5
Particle size and EE of ACDRNPs Cmax µg/ml 3.99
53. 50 5.69
42
Tmax h 3.00
T1/2 h 3.85
All the predicted response lie within 95% CI, and gave
42. 00 5.42
1.00 5
42.83 54.00 65.16 76.33 87. 50 -3 -2 -1 0 1 2 3
0.81 4
0.63 3
X: Actual X: Lambda MRT h 8.02
correlation coefficient value near to 0.999.Hence,
0.44 2 Y : Predic ted Y: Ln(ResidualSS)
B: Stabilizer A : Drug:polymer
0.25 1
AUC(0-t) µg/ml h 5.78
Fig. 3D-Response and Model diagnostic plots of Encapsulation efficiency

The "Pred R-Squared" of 0.5463 is in reasonable agreement with the "Adj R-Squared" of 0.7258. "Adeq Precision"
predicted responses are validated AUC(0-∞) µg/ml h 36.91

measures the signal to noise ratio. A ratio greater than is desirable. Here ratio of 9.487 indicates an adequate signal.
AUMC µg/ml h 289.02
This model can be used to navigate the design space.
oftware
ANOVA Equation: EE=+63.03+10.55 * A-9.66*
h 0.17
Scale up of nanoformulation: Optimized batches
Design-Expert® Sof tware
Predicted vs . Actual
Design-Expert® Software Ka
PDI
PDI Box-Cox Plot for Power Trans form s
ymer Color points by value of

h 0.18
0.24 Lambda
PDI: Current = 1 12.90

0.29
0.22 Best = 1.29 Kel

were formulated and subjected to spray drying

Low C.I. = 0.62
0 High C.I. = 2

0.215 0.18
Recommend transform:
None
(Lambda = 1)
7.67
Vd L 1.33
Fig. In vivo absorption
process
k = 0.0022
(used to make
0.14 response values
PD I

0.12 positive) 2.44

0.065

0.05 -2.79
-0.01

1.00 5.00
-0.01 -8.02
Stability Studies as per ICH guidelines
Spray drying parameters:
0.81 4.00 -0.00 0.06 0.12 0.17 0.23 -3 -2 -1 0 1 2 3

0.63 3.00
0.44 2.00 X: Ac tual X: Lam bda
B: Stabilizer
0.25 1.00
A : Drug:polymer Y : Predic ted Y : Ln(R es idualSS)
Batch Size: 2.5 L
Fig. 3D-Response and Model diagnostic plots of polydipsersity index
Drying
The "Pred R-Squared" of 0.9454 is in reasonable agreement with the "Adj R-Squared" of 0.9812. "Adeq Precision" temperature:1000C
measures the signal to noise ratio. A ratio greater than is desirable. Here ratio of 41.531 indicates an adequate signal. Outlet Temperature:50 0C
This model can be used to navigate the design space. 30 °C ± 2 °C/65% RH ± 5% RH 40 °C ± 2 °C/75% RH ± 5% RH
Feed Rate: 4ml/min
ANOVA Equation: PDI =+0.22+0.083 * A-0.026* B+0.0096 * A * B-0.12* A2+0.015 * B2
Aspirator:400
Optimization as a function of Particle In vitro Particle In vitro
Updated risk assessment parameters
desirability Time Size PDI Assay Release Time Size PDI Assay Release
oftware
Overlay Plot Fig. effect of spray drying on particle Size
1.00

0 Month 321.1 0.214 99.23 98.55% 0 Month 321.1 0.214 99.23 97.60%
P Size:
EE:
319.214
60.2261 Drug Product Drug: Stabilizer Stirring Mixing
In vitro release studies
0.81 PDI: 0.220006
ymer X1
X2
3.96
0.93 CQA's polymer ratio conc. rate time 1 Month 329.5 0.245 99.19 97.84% 1 Month 335.1 0.278 100.5 98.60%
PDI: 0.1
0.63

Assay and CU Low Low Low Low

2 month 328.7 0.227 98.56 98.15 2 month 341.2 0.265 99.78 98.4
EE: 70

CDR Low Low Low Low

0.44 3 month 335.7 0.236 98.47 95.69 3 month 319.5 0.284 99.56 99.06
Particle Size Low Low Low Low

0.25
PDI Low Low Low Low 6 Month 339.5 0.265 98.01 96.15 6 Month 374.1 0.301 99.23 97.4
1.00 2.00 3.00 4.00 5.00

EE Low Low Low Low

X1: A: D rug:poly mer
X2: B: Stabilizer
Working Space ACDRNPs Showed stability over period of 6 months
as per ICH guideline

Establishment of level A of in vitro in vivo correlation Conclusion References

1. Amidon, G.L., et al. Pharmaceutical
 Significant point to point relationship
was observed with regression Research 12, 413–420, 1995
coefficient (R2) of 0.988 and slope 2. C.A. Lipinski, F. Lombardo, B.W. Dominy, P.J.
approaching toward unity, indicating a
Atorvastatin calcium delayed release nanoparticles were successfully prepared and scaled up using QbD Feeney, Advanced Drug Delivery Reviews 46
close correlation between the in vitro approach. (2001) 3–26.
release rates and in vivo absorption of  Five fold increase in bioavailability was observed as compared to marketed formulation and Plain AC
the drug.
The prediction error of the Cmax and suspension. Acknowledgement
AUC of the correlation were found to Successful level A correlation was established on the basis of in-vitro and in- vivo release The authors are grateful to University
be 9.62% and 12.02%. The low Nanoformulation showed stability over a period of six months as per ICH guidelines for the parameters Grants Commission and AICTE for the
prediction error indicates the reliability financial assistance provided for the
of model towards carrying out like particle size, PDI, Assay/Drug content, In vitro release. research work.
predictions; hence it can be selected as
a bio relevant tool to screen the best
IVIVC model linear regression plot