Preparation, Characterization, and in vivo Evaluation of NK4-Conjugated Hydroxycamptothecin-Loaded Liposomes

Figures & data

Figure 1 Schematic of cellular uptake of NK4-HCPT-Lips.

Table 1 Characterization of the Liposomes (n=3)

Sample	Z-Average Size (nm)	Zeta Potential (mV)	PDI	EE (%)	DL (%)
HCPT-Lips	152.9±3.8	−25.7±3.5	0.189±0.09	83.6±3.3	3.19±0.28
NK4-HCPT-Lips	155.6±2.6	−24.8±3.3	0.166±0.07	82.5±2.4	3.01±0.23

Note: Data presented as mean ± standard deviation (n=3).

Abbreviations: HCPT-Lips, Hydroxycamptothecin liposomes; NK4-HCPT-Lips, NK4-modified hydroxycamptothecin liposomes; PDI, polydispersity index; EE, Encapsulation efficiency; DL, drug loading.

Figure 2 Characterization of liposomes.

Notes: (A) TEM image of HCPT-Lips. (B) TEM image of NK4-HCPT-Lips. (C) Size distribution of NK4-HCPT-Lips. (D) Zeta potential of NK4-HCPT-Lips.

Figure 3 In vitro release profiles (37°C, PBS, pH 7.4, n=3).

Note: Data presented as mean ± standard deviation (n=3).

Abbreviations: HCPT-Lips, Hydroxycamptothecin liposomes; NK4-HCPT-Lips, NK4-modified hydroxycamptothecin liposomes.

Table 2 Stability Data of Liposomes (n=3)

Time (Days)	HCPT-Lips		NK4-HCPT-Lips
	Z-Average Size (nm)	EE (%)	Z-Average Size (nm)	EE (%)
7	153.3±2.5	82.3±1.3	156.1±1.3	82.0±1.2
15	154.2±2.9	81.5±0.9	156.3±1.9	81.4±1.1
30	155.5±1.7	81.2±1.1	157.5±2.8	80.2±0.7

Note: Data presented as mean ± standard deviation (n=3).

Abbreviations: HCPT-Lips, Hydroxycamptothecin liposomes; NK4-HCPT-Lips, NK4-modified hydroxycamptothecin liposomes; EE, Encapsulation efficiency.

Table 3 The IC50 of HCPT Formulations (n=3)

Formulations	IC50 (µg/mL)
	24 h	48 h
HCPT-Inject	11.24±0.24	10.66±0.74
HCPT-Lips	21.75±2.73**	7.93±0.39*
NK4-HCPT-Lips	13.74±1.01^##	3.91±0.23^**,##

Notes: *p<0.05, **p<0.01 vs HCPT-Inject; ^##p<0.01 vs HCPT-Lips. Data presented as mean ± standard deviation (n=3).

Abbreviations: HCPT-Lips, Hydroxycamptothecin liposomes; NK4-HCPT-Lips, NK4-modified hydroxycamptothecin liposomes.

Figure 4 In vitro cytotoxicity of HCPT-Inject, HCPT-Lips, or NK4-HCPT-Lips on HepG2 cell for 24 hrs (A) and 48 hrs (B), respectively (n=3).

Notes: *p<0.05, **p<0.01 vs HCPT-Inject; ^#p<0.05, ^##p<0.01 vs HCPT-Lips. Data presented as mean ± standard deviation (n=3).

Abbreviations: HCPT-Lips, Hydroxycamptothecin liposomes; NK4-HCPT-Lips, NK4-modified hydroxycamptothecin liposomes.

Figure 5 Concentration–time curves of HCPT in plasma of mice administered HCPT-Inject, HCPT-Lips, or NK4-HCPT-Lips via tail vein injection.

Note: Data presented as mean ± standard deviation (n=3).

Abbreviations: HCPT-Lips, Hydroxycamptothecin liposomes; NK4-HCPT-Lips, NK4-modified hydroxycamptothecin liposomes.

Table 4 Pharmacokinetic Parameters

Parameter	HCPT-Inject	HCPT-Lips	NK4-HCPT-Lips
Compartment number	One-compartment	Three-compartments	Three-compartments
Cmax (µg/mL)	8.32±1.01	8.40±0.87	8.43±1.23
AUC0-t (µg·h/mL)	1.510±0.121	11.561±1.163**	17.279±2.863^**,#
t1/2(h)	0.349±0.063	1.086±0.152**	1.268±0.196**
MRT (h)	0.289±0.055	2.007±0.210**	3.103±0.356^**,#
CL (L/kg/h)	5.297±0.812	0.665±0.03**	0.373±0.012**^,##

Notes: **p<0.01 vs HCPT-Inject; ^#p<0.05, ^##p<0.01 vs HCPT-Lips. Data presented as mean ± standard deviation (n=3).

Abbreviations: HCPT-Lips, Hydroxycamptothecin liposomes; NK4-HCPT-Lips, NK4-modified hydroxycamptothecin liposomes; C_max, maximum concentration; t_1/2, half-life time; AUC_0–t, area under drug concentration–time curve values (from 0 to time t); MRT, mean residence time; CL, clearance.

Figure 6 Tissues distribution.

Notes: HCPT concentration in heart (A), liver (B), spleen (C), lung (D), and kidney (E) of mice at various time points after i.v. administration. Data presented as mean ± standard deviation (n=3).

Abbreviations: HCPT-Lips, Hydroxycamptothecin liposomes; NK4-HCPT-Lips, NK4-modified hydroxycamptothecin liposomes.