Carfilzomib and Paclitaxel Co-Loaded Protein Nanoparticles an Effective Therapy Against Pancreatic Adenocarcinomas

Figures & data

Figure 1 TEM image and particle size analysis of drug/human serum albumin (HSA) nanoparticles (NPs).

Table 1 50% Inhibitory Concentration (IC₅₀) Values of Different Combinations of Carfilzomib and Paclitaxel

Weight Ratio Combination	IC50 (ng/mL)
	1: 0	1: 2	1: 1	2: 1	0: 1
CFZfree: PTXfree	8.90	0.25	1.39	1.00	0.44
Sb-CFZ: Sb-PTX	8.12	0.03	0.37	0.51	0.47
CFZ/HSA: PTX/HSA	7.89	0.01	0.07	0.66	0.86

Abbreviations: Sb, solvent-based; CFZ, carfilzomib; PTX, paclitaxel; HSA, human serum albumin.

Figure 2 Characterization of the synergistic activity of combined carfilzomib (CFZ) and paclitaxel (PTX) treatment at different weight ratios.

Figure 3 Drug release profiles of carfilzomib (CFZ, A) and paclitaxel (PTX, B). *p<0.05 and **p<0.005.

Table 2 Maximum Tolerance Dose Study for Various Combination Ratios of Carfilzomib and Paclitaxel on BALB/c Mice (n = 4)

Formulations	Single-Dose Study (Days 0)	Multi-Dose Study (Days 0, 1)
Sb-CFZ (mg/kg)	5	2.5
CFZ/HSA (mg/kg)	17.5	5
Sb-PTX (mg/kg)	20	12.5
PTX/HSA (mg/kg)	300	150
Sb-CFZ+Sb-PTX (1:2, mg/kg)	3.75:7.5	<2.5:5
CFZ/HSA+PTX/HSA (1:2, mg/kg)	10:20	5:10
CFZ/PTX/HSA (1:2, mg/kg)	10:20	5:10

Abbreviations: Sb, solvent-based; CFZ, carfilzomib; PTX, paclitaxel; HSA, human serum albumin.

Table 3 Pharmacokinetic Parameters of Carfilzomib Obtained from a Single Intravenous Bolus Administration of Various Formulations (Equivalent to 5 mg/kg Carfilzomib)

Parameter	Sb-CFZ	CFZ/HSA	Sb-CFZ+ Sb-PTX	CFZ/HSA+ PTX/HSA	CFZ/PTX/HSA
C0 (µg/mL)	10.90±3.37	127.86±97.81	11.73±6.43	4.79±1.69	6.35±3.24
Cmax (µg/mL)	3.87±0.63	9.93±2.69	4.2 ±1.33	0.97±0.11	4.30±1.25
T1/2, initial (min)	12.72±2.14	15.19±1.38	11.02±8.98	9.14±2.93	10.00±4.08
T1/2, terminal (h)	1.74±0.34	2.14±0.35	0.53±0.74	0.69±0.42	1.79±0.80
AUC0-1h (h·µg/mL)	0.222±0.034	3.337±4.306	0.241±0.056	0.055±0.009	0.537±0.451
AUC0-24h (h·µg/mL)	0.243±0.030	3.352±4.304	0.248±0.067	0.082±0.018	0.687±0.662
AUC0-infinity (h·µg/mL)	0.243±0.030	3.353±4.304	0.249±0.067	0.082±0.018	0.689±0.661
CL (L/h/kg)	20.78±2.72	4.64±4.32	21.13±5.70	63.29±14.40	12.16±7.83
V (L/kg)	51.53±4.33	12.91±10.63	12.74±16.21	64.98±45.38	37.34±29.31

Abbreviations: Sb, solvent-based; CFZ, carfilzomib; PTX, paclitaxel; HSA, human serum albumin; C₀, initial plasma concentration; C_max, maximum plasma concentration; T_1/2,initial, initial half-life; T_{1/2,terminal,} terminal half-life; AUC, areas under plasma concentration–time curve values for the period of from t=0 to t=1 h, t=0 to 24 h, and t=0 to infinity; CL, plasma clearance; V, volume distribution.

Table 4 Pharmacokinetic Parameters of Paclitaxel Obtained from a Single Intravenous Bolus Administration of Various Formulations (Equivalent to 10 mg/kg Paclitaxel)

Parameter	Sb-PTX	PTX/HSA	Sb-CFZ + Sb-PTX	CFZ/HSA + PTX/HSA	CFZ/PTX/HSA
C0 (µg/mL)	49.47±15.40	23.12±10.94	64.14±15.19	12.57±1.78	13.14±7.70
Cmax (µg/mL)	30.57±8.23	8.92±2.10	29.13±3.62	5.64±0.55	8.21±3.05
T1/2,initial (min)	22.66±1.48	25.77±1.79	23.65±1.05	26.32±5.19	21.90±0.22
T1/2,terminal (h)	7.31±1.84	12.89±0.65	9.08±1.92	13.37±1.41	6.54±0.60
AUC0-2 (h·µg/mL)	4.624±1.140	1.640±0.335	5.074±1.306	1.121±0.288	1.190±0.323
AUC0-24 (h·µg/mL)	6.171±2.018	2.511±0.497	7.004±2.082	1.958±0.824	1.713±0.520
AUC0-infinity (h·µg/mL)	6.216±1.993	2.579±0.522	7.065±2.135	2.056±0.809	1.788±0.563
CL (L/h/kg)	1.71±0.48	3.98±0.77	1.52±0.54	5.32±1.56	6.08±2.31
V (L/kg)	18.83±9.46	73.54±10.72	19.06±2.93	104.75±37.10	58.20±25.90

Note: All parameters are defined in the footnotes to Table 3.

Figure 4 Plasma concentration–time curves of carfilzomib (CFZ, A) and paclitaxel (PTX, B) after intravenous administration at respective doses of 5 and 10 mg/kg to rats.

Figure 5 (A) MIA PaCa-2 tumor growth curve. (B) Tumor inhibition rates. (C) Tumor weights. (D) Body weight changes in tumor-bearing mice.

Dosing time points. *p<0.05 and **p<0.005.

Figure 6 Tissue distributions of carfilzomib (CFZ) and paclitaxel (PTX) at 2 (A and C) and 8 h (B and D), respectively, after intravenous administration of solvent-based (Sb)-CFZ, Sb-PTX, CFZ/human serum albumin (HSA) nanoparticles (NPs), PTX/HSA NPs, Sb-CFZ+Sb-PTX (1:2), CFZ/HSA NPs+PTX/HSA NPs (1:2), or CFZ/PTX/HSA NPs (equivalent to 5 mg/kg CFZ and 10 mg/kg PTX in each mouse). *p<0.05.