Honokiol/Magnolol-Loaded Self-Assembling Lecithin-Based Mixed Polymeric Micelles (lbMPMs) for Improving Solubility to Enhance Oral Bioavailability

Figures & data

Figure 1 Chemical structures of (A) honokiol and (B) magnolol.

Table 1 Preliminary Screening of Mixed Polymeric Micellar Formulations with Various Ratios of Different Amphiphiles (NaDOC, PP123, PF68, PF87, PF127, PF108, and PL121, CRH40 and CELP) and Lecithin

Amphiphile (HLB)	H/M:Lecithin:Amphiphile
	1:0:5	1:1:5	1:0:10	1:1:10	1:0:20	1:1:20
NaDOC (16)	1300.6 ± 316.48^a,c (1.177 ± 0.832)^b	227.8 ± 45.86 (0.631 ± 0.238)	169.6 ± 152.14 (5.877 ± 11.17)	215.7 ± 131.03 (1.211 ± 0.289)	234.7 ± 152.33 (0.456 ± 1.385)	636.3 ± 286.29 (0.035 ± 0.97)
PF68 (>24)	346.2 ± 8.48 (0.091 ± 0.124)	986.9 ± 484.23 (1.461 ± 0.068)	105.6 ± 5.91 (0.129 ± 0.041)	641.5 ± 5.39 (0.495 ± 0.149)	103.9 ± 6.35 (1.26 ± 0.335)	448.2 ± 26.41 (0.655 ± 0.066)
PF108 (>24)	29.9 ± 3.11 (0.675 ± 0.159)	746.7 ± 50.79 (0.603 ± 0.139)	35.1 ± 3.16 (0.762 ± 0.364)	392.7 ± 11.01 (0.546 ± 0.067)	30.2 ± 5.16 (0.725 ± 0.364)	253.9 ± 3.14 (0.466 ± 0.1)
PF87 (>24)	21.8 ± 6.42 (0.275 ± 1.492)	701.1 ± 134.1 (0.816 ± 0.258)	191.3 ± 126.22 (23.08 ± 33.43)	882.2 ± 14.06 (0.211 ± 0.099)	28.4 ± 6.31 (2.124 ± 0.488)	1054 ± 267.64 (0.033 ± 1.068)
PF127 (18–23)	30.9 ± 2.73 (0.558 ± 0.364)	2243 ± 1819 (1.313 ± 0.258)	29.2 ± 1.6 (1.26 ± 0.35)	772.5 ± 101.9 (1.51 ± 0.07)	47.7 ± 47.9 (1.379 ± 1.508)	854.6 ± 336.8 (1.621±0.068)
PL121 (1–7)	≥3000	≥3000	≥3000	≥3000	≥3000	≥3000
PP123 (7–9)	1181.6 ± 51.21 (1.305 ± 0.028)	371.7 ± 12.77 (1.033 ± 0.016)	120.3 ± 9.87 (0.343 ± 0.297)	149.2 ± 1.49 (0.686 ± 0.037)	122.9 ± 5.89 (0.552 ± 0.091)	100 ± 7.12 (0.899 ± 0.062)
CRH40 (14.3)	198.6 ± 0.88 (0.466 ± 0.022)	256.3 ± 7.54 (0.87 ± 0.039)	115 ± 1.42 (1.203 ± 0.063)	348.6 ± 2.75 (1.044 ± 0.064)	28.2 ± 14.29 (0.976 ± 3.515)	322.4 ± 17.37 (1.969 ± 0.026)
CELP (13.9)	164.6 ± 1.57 (0.755 ± 0.01)	235 ± 3.24 (0.758 ± 0.029)	20.3±1.03 (0.862 ± 0.634)	302.5 ± 8.194 (1.298 ± 0.042)	16.2 ± 6.45 (1.044 ± 0.273)	98.6 ± 11.15 (1.591 ± 0.101)

Notes: ^aMean size±standard deviation (SD). ^bMean polydispersity index±SD. ^cUnderlining indicates precipitation during 12 h.

Abbreviations: H/M, honokiol-magnolol; HLB, hydrophilic-lipophilic balance.

Figure 2 Particle sizes of honokiol/magnolol micelles formed using different amphiphiles and ratios.

Table 2 Optimization of Honokiol/Magnolol-Loaded Lecithin-Based Mixed Micelles Formed Using NaDOC and PP123 as Amphiphilic Polymers with Lecithin at Different Ratios

Formulations	PS (nm)	PI	ZP (mV)	DL (%)	EE (%)
H/M:L:NaDOC=1:1:5	121.5 ± 1.888	0.159 ± 0.023	−59.8 ± 3.49	14.60	85.16
H/M:L:NaDOC=1:2:5	50.82 ± 3.645	1.000 ± 0.000	−46.9 ± 4.79	18.65	72.53
H/M:L:NaDOC=2:2:5	83.96 ± 1.422	0.178 ± 0.012	−55.5 ± 2.74	21.55	80.83
H/M:L:NaDOC=3:2:5	79.70 ± 1.324	0.182 ± 0.009	−52.8 ± 2.12	24.00	66.68
H/M:L:NaDOC=4:2:5	82.57 ± 0.202	0.244 ± 0.003	−62.6 ± 2.41	39.60	90.76
H/M:L:NaDOC=5:2:5	96.42 ± 0.632	0.236 ± 0.008	−68.6 ±1.76	42.04	84.08
H/M:L:NaDOC=6:2:5	118.4 ± 2.101	0.186 ± 0.022	−63.7 ± 1.23	44.42	96.41
H/M:L:P123=1:0:5	285.0 ± 3.132	0.253 ± 0.038	−3.13 ± 0.10	16.49	98.93
H/M:L:P123=1:1:5	167.7 ± 3.470	0.253 ± 0.015	−2.38 ± 0.09	13.11	91.79
H/M:L:P123=1:0:10	125.1 ± 1.682	0.280 ± 0.017	−1.73 ± 0.08	6.29	69.15
H/M:L:P123=1:1:10	89.47 ± 1.286	0.413 ± 0.010	1.28 ± 0.10	9.81	99.51
H/M:L:P123=1:0:20	69.68 ± 0.904	0.306 ± 0.026	0.10 ± 0.44	5.35	98.75
H/M:L:P123=1:1:20	66.78 ± 1.555	0.312 ± 0.029	1.70 ± 0.25	5.34	98.62

Abbreviations: H-M, honokiol-magnolol; PS, particle size; PI, polydispersity index; DL, drug loading; ZP, zeta potentials; EE, encapsulation efficiency.

Figure 3 TEM micrograph of lbMPMs[NaDOC] and lbMPMs[PP123]. (A) Honokiol/magnolol: lecithin: NaDOC in a ratio of 6:2:5. (B) Honokiol/magnolol: lecithin: PP123 in a ratio of 1:1:10.

Figure 4 Stability of honokiol/magnolol-loaded (lbMPMs[NaDOC]) (A) and lbMPMs[PP123] (B) stored in phosphate-buffered saline (PBS) at 4 °C and at room temperature; lbMPMs[NaDOC]) (C), lbMPMs[PP123] (D) in PBS or FBS at 37 °C.

Figure 5 In vitro release profiles of honokiol (A) and magnolol (B) from/magnolol Honokiol/magnolol: lecithin: NaDOC in a ratio of 6:2:5 (lbMPMs[NaDOC]) and honokiol/magnolol: lecithin: PP123 in a ratio of 1:1:10 (lbMPMs[PP123]).

Table 3 Summary of Pharmacokinetic Parameters for Honokiol Following Intravenous and Oral Administration of Lecithin-Based Mixed Polymeric Micelles (NaDOC and PP123) and Free Honokiol-Magnolol (n=3)

Treatment Group	IVFree	IVNaDOC^a	IVP123^b	OralFree^c	OralNaDOC^a	OralP123^b
Dose (mg/kg)	20	20	20	40	40	40
Cmax (μg/mL)				-^c	0.13±0.08	0.05±0.02
AUC0-72 (h.μg/mL)	24.87±24.29	23.25±13.00	5.11±2.11	-^c	1.19±1.84	0.10±0.13
kel (1/h)	0.81±1.28	0.07±0.09	0.04±0.02	-^c	0.27±0.30	1.48±2.26
t1/2 (h)	7.3±6.1	20.4±14.3	19.2±6.7	-^c	6.3±6.2	2.8±2.5
Tmax (h)				-^c	0.83±1.01	0.33±0.14
V (L/kg)	20.3±19.7	18.8±5.4	96.7±48.9	-^c	688.1±416.8	1144.8±764.4
CL (L/h/kg)	1.4±1.0	1.06±0.91	3.73±1.77	-^c	219.3±246.5	1007.0±1286.8
Fab (%)	100	93.5	20.5	-^c	4.8	0.4

Notes: ^aHonokiol/magnolol: lecithin: NaDOC=6:2:5; ^bHonokiol/magnolol:lecithin:PP123=1:1:10; ^cnon-detectable.

Abbreviations: k_el, elimination rate constant; t_1/2, half-life; T_max, time of maximum concentration observed; C_max, maximum concentration observed; AUC_0-72, area under curve from the beginning to the end point; V, volume of distribution; CL, clearance; F_ab, absolute bioavailability; F_rel, relative bioavailability.

Table 4 Summary of Pharmacokinetic Parameters for Magnolol Following Intravenous and Oral Administration of Lecithin-Based Mixed Polymeric Micelles (NaDOC and PP123) and Free Honokiol-Magnolol (n=3)

Treatment group	IVFree	IVNaDOC^a	IVP123^b	OralFree	OralNaDOC^a	OralP123^b
Dose (mg/kg)	20	20	20	40	40	40
Cmax (μg/mL)				0.14±0.07	0.39±0.19	1.48±0.44
AUC0-72 (h.μg/mL)	15.95±6.82	54.49±20.97	12.25±9.28	1.10±0.84	3.21±4.61	0.98±0.24
kel (1/h)	0.34±0.54	0.06±0.05	0.04±0.01	0.21±0.06	0.31±0.21	1.54±1.25
t1/2 (h)	13.30±10.90	17.27±10.85	16.05±2.98	3.42±1.00	3.30±2.46	1.17±1.44
Tmax (h)				2.00±1.80	0.67±0.72	0.33±0.14
V (L/kg)	30.88±26.32	7.28±2.61	51.14±33.28	301.20±322.02	136.82±106.51	54.47±60.33
CL (L/h/kg)	1.36±0.46	0.34±0.13	2.18±1.19	63.21±70.88	45.86±36.00	36.20±8.82
Fab (%)	100	341.6***	76.8	6.7	20.1***	6.1
Frel (%)				100	291.8***	89.1

Notes: ^aHonokiol/magnolol:lecithin:NaDOC=6:2:5; ^bHonokiol/magnolol:lecithin:PP123=1:1:10. ***Significant (p < 0.005) compared to IV NaDOC; Parameters are defined in the footnotes to Table 3.

Figure 6 Plasma concentration-time curves of honokiol ((A): 0–72h; (B):0–12h) and magnolol ((C): 0–72h; (D): 0–12h) after intravenous administration of lbMPMs[NaDOC], lbMPMs[PP123] and free honokiol-magnolol (20 mg/kg) to rats. Each point represents the mean±SD of three determinations (n=3).

Figure 7 Plasma concentration-time curves of honokiol ((A): 0–72h; (B): 0–8h) and magnolol ((C): 0–72h; (D):0–8h) after oral administration of lbMPMs[NaDOC], lbMPMs[PP123] and free honokiol-magnolol at a single dose of 40 mg/kg to rats. Each point represents the mean±SD of three determinations (n=3).