Formulation and Characterization of Intranasal Drug Delivery of Frovatriptan-Loaded Binary Ethosomes Gel for Brain Targeting

Figures & data

Table 1 Formulation and Characterization of BE Formulations

Formula	PL (w/v)	Ethanol (mL)	PG (w/v)	Ch (mg)	PBS (Up to mL)
	1	0	0	0	45
2	3	0	0	0	45
3	1	20	5	1	45
4	2	20	7.5	1	45
5	3	20	10	1	45
6	4	20	15	1	45
7	1	10	5	1	45
8	1	20	7.5	1	45
9	1	30	10	1	45
10	1	40	15	1	45
11	2	10	5	0.5	45
12	2	20	7.5	1.5	45
13	2	30	10	1.5	45
14	2	40	15	2	45
15	4	10	5	1	45
16	4	20	7.5	1	45
17	4	30	10	1	45
18	4	40	15	1	45
19	3	10	5	1	45
20	3	20	7.5	1	45
21	3	30	10	1.5	45
22	3	40	15	1	45
23	1	10	10	1	45
24	2	20	10	1	45
25	3	30	10	1	45

Abbreviations: PL, Phospholipon^® 90H [PL90H], PBS, Phosphate-buffered saline, Ch, cholesterol, PG, Propylene glycol.

Table 2 Formulation of FVT BE-Loaded Gel from the Optimized Formula (F9)

Formulation	F9 dispersion (mL) (2.5 mg/mL)	Poloxamer 407 (g)	Carbopol 934 (g)	HPMC K100 (g)	DW (mL)
G1	10	14	0.5	–	100
G2	10	14	1.0	–	100
G3	10	14	1.5	–	100
G4	10	14	–	0.5	100
G5	10	14	–	1.0	100
G6	10	14	–	1.5	100
G7	10	14	1.0	1.0	100

Abbreviations: DW, distilled water; HPMC, (Hydroxypropyl) methylcellulose.

Figure 1 Particle size and zeta potential of optimal TVF loaded BE (F9).

Table 3 Characterization of FVT Loaded BE

Formulation	EE%	PS	PDI	ZP
F1	63.98±2.18^a	352.69±5.37^a	0.413±0.01^a	−24.39±0.21^a
F2	69.31±0.98^a	494.83±2.64^b	0.521±0.06^a	−22.59±1.05^a
F3	80.51±0.98^b	200.37±2.64^c	0.294±0.06^a	−32.13±2.16^b
F4	69.82±3.06^a	265.49±2.84^d	0.367±0.03^a	−38.37±0.36^c
F5	59.38±2.14^a	326.49±9.62^e	0.473±0.04^a	−20.46±0.49^a
F6	72.34±2.61^c	264.72±7.34^d	0.196±0.02^b	−43.94±0.12^d
F7	54.29±0.94^d	268.31±9.31^d	0.375±0.01^a	−28.49±2.04^b
F8	69.42±1.52^ac	220.59±6.52^f	0.322±0.06^ab	−31.27±2.31^b
F9	89.34±2.37^e	154.1±4.38^g	0.213±0.05^b	−46.94±1.05^d
F10	75.96±3.16^bc	210.34±1.98^cf	0.487±0.03^a	−26.59±0.05^a
F11	78.35±2.38^bc	254.95±2.35^d	0.203±0.05^b	−38.37±0.07^c
F12	62.54±0.97^a	235.67±10.59^f	0.386±0.04^a	−26.04±0.04^a
F13	49.37±0.35^d	433.92±2.46^h	0.495±0.01^a	−29.38±0.05^b
F14	45.12±3.59^f	376.22±8.37^i	0.512±0.03^a	−39.07±0.28^c
F15	74.39±1.05^bc	147.35±1.39^g	0.418±0.05^a	−22.7 ± 2.3^a
F16	70.34±2.13^ac	234.51±8.31^f	0.229±0.01^b	−44.27±2.03^d
F17	65.39±3.05^ac	295.37±7.26^j	0.324±0.03^a	−33.15±1.06^b
F18	64.51±2.03^a	235.67±6.29^f	0.367±0.05^a	−36.41±1.34^c
F19	69.42±1.52^ac	220.59±6.52^f	0.322±0.06^ab	−31.27±2.31^b
F20	60.53±2.37^ad	394.28±4.38^i	0.413±0.05^a	−32.13±2.16^b
F21	55.96±3.16^d	210.34±1.98^f	0.487±0.03^a	−26.59±0.05^a
F22	68.35±2.38^ac	254.95±2.35^cd	0.361±0.05^a	−28.37±0.07^b
F23	62.54±0.97^a	235.67±10.59^f	0.386±0.04^a	−26.04±0.04^a
F24	49.37±0.35^df	433.92±2.46^h	0.495±0.01^a	−29.38±0.05^b
F25	75.12±3.59^bc	295.22±8.37^j	0.268±0.03^b	−39.07±0.28^c
p-value	<0.0001	<0.001	<0.001	<0.001

Notes: A column with different letters indicates significant differences (≤0.05)‎ [^{a vs b, c, d, e, f, g, h, i, j} ≤0.05; ^{b vs c, d, e, f, g, h, i, j}≤0.05; ^{c vs d, e, f, g, h, i, j}≤0.05; ^{d vs e, f, g, h, i, j}≤0.05; ^{e vs f, g, h, i, j}≤0.05; ^{f vs g, h, i, j}≤0.05; ^{g vs h, i, j}≤0.05; ^{h vs i, j}≤0.05; ^{i vs j}≤0.05].

Abbreviations: EE, entrapment efficiency‎; PS, particle size; PDI, polydispersity index; ZP, zeta potential.

Figure 2 Entrapment efficiency (%) of various BE formulas [a column with different letters (a, b, c, d, e, and f) indicates significant differences (≤0.05)‎].

Figure 3 Particle size (nm) of various BE formulas [a column with different letters (a, b, c, d, e, f, g, h, i, and j) indicates significant differences (≤0.05)‎].

Figure 4 Polydispersity index of various BE formulas [a column with different letters (a, and b) indicates significant differences (≤0.05)‎].

Figure 5 Zeta potential of various BE formulas [a column with different letters (a, b, c, and d) indicates significant differences (≤0.05)‎].

Table 4 Physical Evaluation of FVT BE Gel Formulations

	P407 (w/v%)	C934 (w/v%)	HPMC	Visual Appearance	pH	Gelation Temperature °C	Spreadability (g.cm/sec)	Drug Content (%)
G1	14	0.5	–	Milky appearance	5.8±0.01^a	33.9±0.21^a	9.68±1.31^a	92.57±0.21^a
G2	14	1.0	–	Milky appearance	5.3±0.03^b	33.7±0.43^a	12.84±2.43^b	99.82±0.02^b
G3	14	1.5	–	Milky appearance	5.1±0.01^b	31.9±0.13^a	6.57±1.27^a	95.43±0.14^c
G4	14	–	0.5	Milky appearance	5.4±0.02^b	34.2±0.26^a	7.64±0.98^a	98.54±0.37^d
G5	14	–	1.0	Milky appearance	6.0±0.01^c	30.09±1.00^a	6.82±1.34^a	94.21±0.28^e
G6	14	–	1.5	Milky appearance	6.2±0.02^d	28.9±2.30^b	5.37±1.29^c	96.56±0.11^f
G7	14	1.0	1.0	Milky appearance	6.5±0.01^e	35.6±3.21^a	7.81±1.17^a	92.54±0.13^a
p-value	-	-	-	-	<0.001	0.001	<0.001	<0.001

Notes: P407: Poloxamer ‎407 (w/v%), C934: Carbopol 934 (w/v%)‎, HPMC: hydroxypropyl methylcellulose. A column with different letters indicates significant differences (≤0.05)‎ [^{a vs b, c, d, e, f}≤0.05; ^{b vs c, d, e, f} ≤0.05; ^{c vs d, e, f} ≤0.05; ^{d vs e, f} ≤0.05; ^{e vs f}≤0.05.].

Figure 6 DSC Thermograms of 1) Pure Drug (FVT) [1-DSC], 2) Lipid (PL90H) [2-DSC], 3) Poloxamer 407 [4-DSC], 4) FVT-loaded BE (F9) [5-DSC], 5) FVT-loaded BE gel (G2) [8-DSC].

Figure 7 FTIR spectral studies of 1) Pure Drug (black line; line-11), 2) phospholipon 90H‎ (red line; line-21), 3) ‎Ethanol (white, blue; line-31), 4) propylene glycol (light green; line-41), 5) Poloxamer 407 (white gray; line-51), 6) F9 (dark red; line-61), 7) Carbopol 934 (dark green; line-71), 8) G2 (dark blue; line 81).

Figure 8 SEM images of optimized FVT-loaded BE F9 (A and B), optimized FVT-loaded BE gel G2 (C and D).

Figure 9 In vitro drug diffusion profiles of FVT solution, G2, and F9 formulation.

Figure 10 Ex vivo permeation study at nasal pH 6.4.

Figure 11 Light photomicrograph of (A) untreated rat epithelium and rat epithelium treated with (B) normal saline pH 6.8, (C) Drug solution, (D) F9 and (E) G2. “m”, mucosa; “c”, underlying cartilage; “o”, lamina propria of the mucosal layer; “v”, blood vessels in lamina propria.

Table 5 Pharmacokinetic Parameters, Brain Targeting Efficiency, and Direct Nose-to-Brain Transport Following Administration of FVT Formulations

Parameters	Tissue/Organ	Formulations of FVT
		FVT Solution	F9	G2
Cmax	Plasma	43.29±13.59	670.51±24.36	452.87±35.67
	Brain	16.46±06.45	118.34±14.29	495.84±1892
Tmax	Plasma	45.62	–	15
	Brain	60.87	34	10
AUC0-360	Plasma	5737.16±184.53	52,684.35±225.34	15,385.46±1231.26
	Brain	1842.84±164.72	14,876.29±94.36	36,484.28±1159.46
AUC ratio		0.32	0.2825	2.3713
Fa		11.96	100	29.47±0.26
Fr		100	-	268.79±0.59
T1/2	Plasma	152.36±12.35	98.37±8.34	15,857±11.56
	Brain	228.43±5.64	133.84±9.03	198.37±8.64
MRT	Plasma	226.07±15.69	116.87±6.59	162.63±10.29
	Brain	331.86±29.64	184.35±5.38	273.84±7.64
DTE		121.84	-	874.18
DTP		18.23	-	89.42

Abbreviations: DTP, brain drug direct transport percentage; MRT, mean residence time; DTE, drug targeting ‎efficiency; Fa absolute bioavailability; Fr, relative bioavailability.