Table 1. QTPP and CQAs for the preparation of NLCs and their justifications.
QTPP
|
Target
|
Justification(s)
|
Dosage form
|
Freeze-dried Nanocarriers
|
Solid dosage form is a preferred choice of the patient because it easily enhances the lymphatic uptake due to its nanosize thereby increases the bioavailability.
|
Dosage type
|
Controlled release
|
Superior absorption is possible with controlled release from NLCs.
|
Route of administration
|
Oral
|
Most preferred route for all including adults and child for the HIV treatment.
|
Drug content
|
85-95 %
|
IT is important to maintain the therapeutic requirement and need for treatment.
|
Drug release
|
More than 85 %
|
Required for effective treatment and therapy inventiveness for improvement of bioavailability of the drug.
|
Pharmacokinetics
|
Maximum Cmax, Tmax and AUC determine performance.
|
The maximum plasma concentration attained versus time provides a higher drug absorption rate and enhance bioavailability.
|
Stability
|
12 months
|
The therapeutic potential of the drug in NLCs depends on particle size. The formulation should not have great variation during storage.
|
Alternate route
|
-
|
NA
|
CQAs
|
Target
|
Justification(s)
|
Particle size
|
Average particle size below 250 nm
|
Reduced particle size and increased surface area enables efficient active movement in permeation through GI epithelial lining. The benefit of reduced size is in the enhancement of solubility and bioavailability.
|
Dispersity index
|
Less than 0.300
|
The dispersity higher than 0.3 indicates the broader dispersity of particles. The dispersity index below 0.3 is acceptable for the nanoformulation.
|
EE
|
Higher
|
Higher EE ensures higher encapsulation and drug loading. The controlled release from the carrier can be obtained for higher therapeutic benefits.
|
Table 2 CCRD for different experimental run and their responses
|
Independent variables
|
Dependent variable (Response)
|
Run
|
Ultrasound amplitude
(A) (%)
|
Lipid concentration
(B) (%)
|
Surfactant concentration (C) (%)
|
Particle size (Y1)
|
PDI (Y2)
|
EE (Y3)
|
1
|
40
|
2
|
0.29
|
191.1±1.01
|
0.311±2.12
|
87.01±1.20
|
2
|
40
|
2.70
|
1
|
213.5±2.05
|
0.204±2.93
|
92.03±2.12
|
3
|
54.14
|
2
|
1
|
164.4±1.03
|
0.362±1.03
|
68.04±0.23
|
4
|
50
|
1.5
|
1.5
|
159.3±1.06
|
0.215±0.32
|
81.10±1.32
|
5
|
40
|
2
|
1
|
187.9±1.23
|
0.209±1.06
|
83.20±1.43
|
6
|
30
|
2.5
|
1.5
|
227.4±2.01
|
0.214±0.34
|
94.30±3.23
|
7
|
40
|
2
|
1
|
190.1±2.03
|
0.186±1.32
|
84.12±1.32
|
8
|
25.85
|
2
|
1
|
247.4±1.13
|
0.288±3.02
|
80.01±3.12
|
9
|
40
|
2
|
1
|
184.8±1.04
|
0.209±1.43
|
81.10±0.32
|
10
|
40
|
1.29
|
1
|
172.1±0.53
|
0.135±0.23
|
91.04±0.43
|
11
|
40
|
2
|
1
|
202.2±0.94
|
0.144±0.34
|
74.03±1.04
|
12
|
40
|
2
|
1.70
|
162.2±1.04
|
0.241±1.07
|
92.10±1.23
|
13
|
30
|
1.5
|
0.5
|
161.1±2.32
|
0.221±1.22
|
84.05±1.42
|
14
|
50
|
2.5
|
0.5
|
228.9±1.32
|
0.188±0.43
|
94.03±0.33
|
15
|
40
|
2
|
1
|
189.5±0.34
|
0.231±1.08
|
84.20±0.12
|
Mean ± SD, (n=3); Lower values (-1) were represented by 30 % (A), 1.5 % (B), 0.5 % (C); Middle values (0) were represented by 40 % (A), 2 % (B), 1 % (C); and Higher values (+1) were represented by 50 % (A), 2.5 % (B), 1.5 % (C). Minimum alpha values (axial points) (-α) were represented by 25.85 %, 1, 29 % and 0.29 %; Maximum alpha values (axial points) (+α), were represented by 54.14 %, 2.70 %, and 1.70 %
Table 3. Results for confirming optimization of formulation (RTV-NLCs).
Experimental variables
|
|
|
|
|
Ultrasound amplitude (%)
|
Lipid conc. (%)
|
Surfactant conc. (%)
|
Responses
|
Predicted values
|
Observed values
|
% Bias
|
40
|
1.82
|
1.43
|
Particle size
|
189.86
|
187.23±1.03
|
1.3
|
|
|
|
PDI
|
0.110
|
0.219±0.02
|
99
|
|
|
|
EE
|
95.96
|
92.01±1.21
|
4.11
|
Table 4. Stability study for optimized RTV loaded NLCs.
Values are expressed as mean ± SD, (n=3) and statistical significant difference (p-value < 0.05).
Table 5. Apparent permeability through intestine in rat gut sac model and pharmacokinetic profile of RTV-NLCs, PM, and RTV-suspension in rat plasma.
Formulations
|
Apparent permeability (Papp)
|
Optimized RTV-NLCs
|
18.32
|
RTV-PM
|
7.825
|
RTV-Suspension
|
6.25
|
Table 6. Pharmacokinetic profile of RTV-NLCs, PM and RTV-suspension in rat plasma.
Pharmacokinetic parameters
|
RTV-Suspension
|
RTV-PM
|
RTV-NLCs
|
Tmax (h)
|
2
|
4
|
4
|
Cmax (µg/mL*h)
|
0.165 ±0.12
|
0.255±0.03
|
0.472 ±0.21
|
AUC 0-t (µg/mL*h)
|
2.40 ±0.24
|
2.99±2.32
|
4.44 ±0.11
|
AUC 0-∞ (µg/mL*h)
|
2.67 ±0.02
|
3.90±2.10
|
3.44 ±2.82
|
MRT
|
10.15±1.25
|
16.28±1.12
|
11.62±0.21
|
Vz/F
|
1418.5±1.32
|
1695.24±1.01
|
802.04±0.21
|
Cl/F
|
153.38±0.01
|
102.51±0.21
|
78.26±0.12
|
Relative bioavailability
|
-
|
-
|
286
|
Values are expressed as mean±SD, (n=3) and statistical data were analyzed by one-way ANOVA and followed by Dunnett’s tests, with significance level p < 0.05. Cmax: maximum concentration, AUC: Area under curve, MRT: mean residence time; Tmax: time to attain maximum concentration.