Binary Nanodrug-Delivery System Designed for Leukemia Therapy: Aptamer- and Transferrin-Codecorated Daunorubicin- and Luteolin-Coloaded Nanoparticles

Figures & data

Figure 1 Scheme (A) and TEM images (B) of AP/Tf-Drn/Lut NPs.

Note: AP/Tf-Drn/Lut NPs were nanosized with some ligands on the spherical surface.

Table 1 Characterization of nanodrug-delivery systems (means ± SD, n=3)

Formulations	Particle size (nm)	PDI	ζ-potential (mV)	Drn		Lut
				EE (%)	LC (%)	EE (%)	LC (%)
AP/Tf-Drn/Lut NPs	187.3±5.3	0.142±0.019	−25.4±2.6	88.7±3.9	5.2±0.5	85.9±4.2	2.1±0.4
AP-Drn/Lut NPs	186.7±4.7	0.139±0.023	−19.2±2.1	87.5±3.8	5.9±0.6	86.7±3.7	2.4±0.6
Tf-Drn/Lut NPs	188.3±4.5	0.126±0.016	−17.5±1.8	86.5±4.1	5.7±0.6	88.3±3.9	2.2±0.5
AP-Drn NPs	91.5±2.8	0.112±0.011	+18.9±1.7	89.4±4.4	11.8±1.1	/	/
Tf-Lut NPs	88.7±2.5	0.128±0.014	−38.9±3.1	/	/	86.5±3.6	4.6±0.7
AP/Tf NPs	187.8±5.1	0.147±0.026	−37.6±2.9	/	/	/	/

Figure 2 Changes in particle size (A) and EE (B) analyzed in PBS and culture medium (FBS). In vitro drug-release behavior of Drn (C) or Lut (D) from nanosystems evaluated by dialysis.

Notes: Sustained drug-release patterns were found for all the samples tested. Data presented as means ± SD, n=3.

Table 2 Cellular uptake percentages (means ± SD, n=8)

Formulations	1 h	24 h
AP/Tf-Drn/Lut NPs	73.1±3.6	65.8±3.3
AP-Drn/Lut NPs	59.5±3.2	55.6±2.8
Tf-Drn/Lut NPs	57.1±2.9	54.2±3.1
AP-Drn NPs	60.2±3.3	53.1±2.6
Tf-Lut NPs	58.4±2.7	52.5±2.8
AP/Tf NPs	74.3±3.5	64.7±3.2

Table 3 CI₅₀ values of AP/Tf-Drn/Lut NPs when different Drn:Lut weight ratios were applied (means ± SD, n=8)

Formulations	Drn:Lut (w:w)	IC50 of Drn (μM)	IC50 of Lut (μM)	CI50
AP-Drn NPs	/	0.93±0.09	/	/
Tf-Lut NPs	/	/	1.16±0.12	/
AP/Tf-Drn/Lut NPs	5:1	0.79±0.08	1.06±0.11	0.987
AP/Tf-Drn/Lut NPs	5:2	0.56±0.05	0.22±0.03	0.792
AP/Tf-Drn/Lut NPs	1:1	0.45±0.04	0.45±0.04	0.872
AP/Tf-Drn/Lut NPs	2:5	0.29±0.03	0.73±0.09	0.941
AP/Tf-Drn/Lut NPs	1:5	0.18±0.02	0.90±0.10	0.969

Figure 3 Cytotoxicity of AP/Tf-Drn/Lut NPs and other formulations evaluated with MTS assays.

Notes: Cytotoxicity of AP/Tf-Drn/Lut NPs was remarkably higher than single ligand–decorated NPs, single drug–loaded NPs, and free drugs. Data presented as means ± SD, n=6. *P<0.05.

Figure 4 In vivo AML therapy efficiency: Tumor size (A) and body weight (B).

Notes: AP/Tf-Drn/Lut NPs exhibited the most remarkable AML therapy efficiency compared with single ligand–decorated, single drug–loaded and free-drug groups. Data presented as means ± SD, n=8. *P<0.05.

Table 4 Pharmacokinetic parameters for Drn (means ± SD, n=8)

Parameters	Unit	AP/Tf-Drn/Lut NPs	AP-Drn/Lut NPs	Tf-Drn/Lut NPs	Free Drn/Lut
Cmax	L/kg/h	55.36±3.21*	42.31±2.98*	40.55±2.74*	29.83±2.88
t½	h	12.37±0.78*	9.72±0.64*	8.84±0.53*	1.89±0.31
AUC0–t	mg/L/h	659.72±19.56*	512.33±17.14*	488.75±21.16*	256.81±9.18
AUC0–∞	mg/L/h	662.31±20.05*	519.64±19.47*	493.23±22.44*	404.73±9.26

Note: *P<0.05 compared with free Drn/Lut.

Abbreviations: C_max, plasma drug peak concentration; t_½, half-life; AUC_0–t, area under curve of time 0 to last time point; AUC_0–∞, area under curve of time 0 to maximum.

Table 5 Pharmacokinetic parameters for Lut (mean ± SD, n=8)

Parameters	Unit	AP/Tf-Drn/Lut NPs	AP-Drn/Lut NPs	Tf-Drn/Lut NPs	Free Drn/Lut
Cmax	L/kg/h	35.47±3.18*	28.11±2.36*	26.59±2.95*	18.31±2.12
t½	h	8.98±0.58*	5.46±0.41*	5.31±0.34*	1.51±0.29
AUC0–t	mg/L/h	431.25±11.38*	311.26±8.34*	289.86±7.65*	198.63±4.59
AUC0–∞	mg/L/h	439.35±12.24*	317.64±11.35*	295.61±6.96*	202.34±5.13

Note: *P<0.05 compared with free Drn/Lut.

Abbreviations: C_max, peak plasma drug concentration; t_½, half-life; AUC_0–t, area under curve of time 0 to last time point; AUC_0–∞, area under curve of time 0 to maximum.

Figure 5 In vivo Drn (A and C) and Lut (B and D) distribution in tissue after 1 h (A and B) and 48 h (C and D) of drug administration. *P<0.05.

Notes: AP/Tf-Drn/Lut NPs showed higher tumor-tissue distribution than single ligand–decorated AP-Drn/Lut NPs, Tf-Drn/Lut NPs, and free Drn/Lut. Data presented as means ± SD, n=8. *P<0.05.