Liver Targeting of Daclatasvir via Tailoring Sterically Stabilized Bilosomes: Fabrication, Comparative In Vitro/In Vivo Appraisal and Biodistribution Studies

Figures & data

Table 1 Observed Box–Behnken Experimental Runs of DAC-Loaded BILS with Their Actual and Predicted Experimental Values of Y₁, Y₂, and Y₃

Runs	Independent Variables			Dependent Variables						PDI
	A	B	C	Y1 (Mean ± SD)		Y2 (Mean ± SD)		Y3 (Mean ± SD)
				Actual	Predicted	Actual	Predicted	Actual	Predicted
1	3:1	15	30	312±16	304.8	87±3.39	87.5	43±1.24	43.2	0.30
2	3:1	0	10	395±19	402.5	85±2.65	85	47±2.02	45.7	0.30
3	7:1	30	10	480±15	472.5	95±1.79	95	33±3.61	32.7	0.20
4	5:1	0	5	408±11	402.6	90±1.73	90	39±1.15	41.7	0.20
5	5:1	15	10	310±11	316.6	97±1.40	96.3	39±2.35	39.2	0.30
6	5:1	30	30	290±10	295.0	95±1.78	95	35±1.50	36.7	0.20
7	7:1	0	10	470±17	468.0	89±1.93	89.5	40±2.14	40.2	0.50
8	5:1	30	5	390±14	390.0	97±1.94	97.5	35±2.74	34.2	0.20
9	3:1	30	10	370±26	371.7	89±1.92	88.5	37±4.01	38.2	0.30
10	5:1	15	10	320±26	316.6	97±1.37	96	41±3.05	39.2	0.30
11	7:1	15	30	385±14	387.0	92±1.41	92	38±2.00	37.7	0.30
12	7:1	15	5	475±13	482.0	93±1.21	92.5	35±2.25	35.2	0.27
13	3:1	15	5	400±17	397.8	86±2.47	86	41±2.83	40.7	0.29
14	5:1	0	30	310±8	309.6	94±2.24	93.5	44±4.00	44.2	0.29
15	5:1	15	10	320±18	316.6	95±2.89	96.3	41±4.11	39.2	0.30

Notes: A: ratio of PL:SDC; B: cholesterol amount (mg); C: sonication time (min); Y₁: particle size (nm); Y₂: EE (%); Y₃: drug release (%).

Figure 1 Response surface plots for the effect of ratio of PL:SDC (A), CH amount (B), and sonication time (C) on (Y₁) particle size, (Y₂) EE%, and (Y₃) % drug release.

Figure 2 Transmission electron micrograph of unPEG-BILS (A) and PEG-BILS (B) optimum formulation.

Table 2 Pharmacokinetic Parameters of DAC Suspension, DAC-unPEG-BILS, and DAC-PEG-BILS in Rat Plasma Following Oral Administration (n=6)

Parameters	DAC Suspension	DAC-UnPEG-BILS	DAC-PEG-BILS
t½ (h)	11.40±0.14	13.80±0.98^a	15.46±0.35^a, b
Tmax (h)	1.0±0.16	2.0±0.44^a	2.0±0.39^a
Cmax (ng.mL^−1)	1300±122.12	1333±541.27	1468±115.41^a, b
AUC0–24 (h.ng.mL^−1)	14,892±437.61	19,238±466.61^a	22,943±212.39^a, b
AUMC0–24 (ng.h^−2.mL^−1)	113,350±437.61	146,037±527.32^a	183,496±374.58^a, b
MRT0–24 (h)	7.6±0.2	7.6±0.35	8.0±0.18^a, b
CL (L.h^−1)	0.8±0.02	0.6±0.05^a	0.5±0.05^a, b

Notes: Values are means ± SD. Using one-way ANOVA followed by Tukey’s post-hoc test, ^ap<0.05 vs DAC suspension; ^bp<0.05 vs DAC-unPEG-BILS.

Figure 3 Mean plasma concentration–time curves of DAC in rats after oral administration of DAC suspension, DAC-unPEG-BILS, and DAC-PEG-BILS at a dose equivalent to 60 mg/kg of DAC (n=6).

Table 3 Pharmacokinetic Parameters of DAC Suspension, DAC-unPEG-BILS, and DAC-PEG-BILS in Rat Liver Following Oral Administration (n=6)

Parameters	DAC Suspension	DAC-UnPEG-BILS	DAC-PEG-BILS
Cmax (ng.mL^−1)	1205±13.32	1596±20.39^a	1938±14.41^a, b
Tmax (h)	2±0.09	2.7±0.21^a	3±0.17^a, b
AUC0–24(h.ng.mL^−1)	9673±13.69	20,698±16.32^a	29,895±19.39^a, b
t½ (h)	11±0.14	13.46±0.14^a	15.66±0.14^a, b
CL (L.h^−^1)	1.20±0.11	0.57±0.03^a	0.4±0.04^a, b

Notes: Values are means ± SD. Using one-way ANOVA followed by Tukey’s post-hoc test: ^ap<0.05 vs DAC suspension; ^bp<0.05 vs DAC-unPEG-BILS.

Table 4 Hepatic Specific Targeting Parameters After Oral Administration of DAC Suspension, DAC-unPEG-BILS, and DAC-PEG-BILS in Rats (n=6)

Parameters	DAC Suspension	DAC-UnPEG-BILS	DAC-PEG-BILS
AUC0–24(ng.h.mL^−1)	9673±13.69	20,698±16.32	29,895±19.39
Re	–	2±0.07	3±0.04
Te	0.64±0.01	1.07±0.06	1.30±0.05
Ce	–	1.32±0.03	1.61±0.09

Abbreviations: AUC, area under the plasma concentration–time curve; Ce, peak concentration ratio; Re, intake rate; Te, targeting efficacy; Re, liver AUC₀₋₂₄ BILS/liver AUC₀₋₂₄ suspension; Te, AUC_liver/AUC_plasma; Ce, liver (C_max)BILS/liver (C_max)_Suspension.