Inhalable core-shell hybrid lipid-protein nanocomposites for tumor-targeted co-delivery of genistein and tretinointo lung carcinoma

Abstract

Lung cancer is the most prevalent type of cancer and the leading cause of cancer-related death. The Inhalation delivery is more effective administration route in treatment lung cancer than traditional methods due to local accumulation and low toxicity for chemotherapeutic drugs. Polymer–lipid hybrid nanoparticles exhibits complementary characteristics of both polymeric hybrid nanoparticles and lipid nanoparticles like high drug loading efficiency, high stability and biocompatibility, controlled release properties, and increased drug stability. Proteins represent good raw materials since they have the advantages of synthetic polymers together with the advantages of absorbability, low toxicity of the degradation, greater stability during storage and in vivo and their ease to scale up during manufacture over other drug delivery systems. Therefore, in this study, inhalable protein-lipid hybrid nanocomposites were prepared for co-delivery of the hydrophobic anticancer drugs genistein and all-trans retinoic acid (ATRA). Nanoparticles were prepared via solvent injection method using glyceryl monostearate as lipid core solubilizing genistein and zein as hydrophobic protein shell solubilizing ATRA. The zein shell was successfully decorated with dual tumor-targeting ligands. Dry powder inhaler for nanoparticles were prepared by spray drying method with different carriers. The mean particle size of dual targeted formulation was 206 ± 10 nm, PDI of 0.2±0.01, zeta potential of 37±5 and genistein entrapment efficiency of 90±2% and tretinoin 98.7±2%. The protein-lipid hybrid nanoparticles exhibited a sustained drug release with 64% of genistein released after 48 hours compared to100% for free drug after 4 hours. The zein shell of nanoparticles were conjugated with the small molecules as biotin and tretinoin by carbodiimide reaction. The dry powder inhaler had high aerodynamic properties as FPF 70%, ED was 97.9 % and MMAD was 2.5 μm. These results indicated that genistein has high entrapment efficiency in the lipid core whereas the protein shell improved the sustained drug release and formulation stability and nanocomposite formulation was deposit in deep lung.

Full text

Abstract
Inhalable protein-lipid hybrid nanocomposites were prepared for co-
delivery of the hydrophobic anticancer drugs genistein and all-trans
retinoic acid (ATRA) for lung cancer treatment. The zein shell was
successfully conjugated with dual tumor-targeting ligands. Dry powder
inhaler for nanoparticles were prepared by spray drying method with
different type of carriers.
Introduction
•Lung cancer is the most prevalent type of cancer and the leading
cause of cancer-related death. The Inhalation delivery is more effective
administration route in treatment lung cancer than traditional methods.
•Polymer–lipid hybrid nanoparticles exhibits complementary
characteristics of both polymeric hybrid nanoparticles and lipid
nanoparticles.
•All-trans retinoic acid affects DNA synthesis, this leads to the cell
arrest before G1 phase. Genistein is potent tyrosine kinases (TKs)
inhibitor. It could inducing arrest cell cycle at G2/M phase.
•There is synergistic effect between two drugs. genistein enhances the
ATRA induced tumor cell differentiation and apoptosis . Also, genistein
reduces the ATRE resistance in cancer cell.
Materials and Methods
lipid-protein core-shell nanoparticles was prepared by solvent injection
method using glyceryl monostearate (GMS) as lipid core solubilizing
genistein with homogenization and heating. zein as hydrophobic
protein shell solubilizing ATRA precipitated on surface of solid lipid
nanoparticles with solvent evaporation method.
Targeting ligands (biotin and tretinoin) Conjugated on protein by
carbodiimide reaction.
Results and discussion
Table.1 Physical characterizations for different formulations.
Table.2 Composition and properties of hybrid lipid protein nanoparticles
Figure 1. Genistien and tretinoin release curve in PBS (7.4PH) with 0.5%
tween 80
Figure 2 .Transmition electron microscope for hybrid nanoparticles
Figure 3. IC50 for TRE and GNS loaded in different formulations on
A549 cell line
Table 3. The spray dried powder characterizations with different
carriers
Table 4. aerodynamic properties (FPF, ED%, MMAD and GSD)for
DPI of different formulations
Figure (4 ,5). Scanning electron microscope for dry powder for F4
References
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Acknowledgments
This work has been funded by STDF (Targeted call
pharmaceutical industries, Grant No. 5731).
Conclusion
These results indicated that hydrophobic drugs have high
entrapment efficiency in the lipid core and zein shell and had high
cytotoxic effect on lung cancer cells . Dry powder nanocomposites
had suitable aerodynamic properties for accumulating nanoparticles
to deep lung.
No. FPF (%) ED (%)
MMAD (µm)
GSD
F1 13.68 % 74.8 % 6.8 1.27
F2 58.74 % 78.14 % 3.28 2.13
F3 54.39 % 70.37 % 3.14 2.13
F4 70.81 % 97.9 % 2.47 2.13
Type of lipid
Lipid amount
(mg) Surfactants P.S. (nm)
Tween 80 (mg)
PC S75 (mg)
GMS 200 500 100 165.9
GMS 100 500 100 159.1
GMS 300 500 100 164.3
GMS 200 1000 200 191.8
GMS 200 250 50 373.5
Geleol 200 500 100 211.5
Biogarpess 200 500 100 295.3
Precirol 200 500 100 245.3
Gelucire 200 500 100 257
zetaPDIPS (nm)formulations
37.40.17165.9non-targeted
nanoparticles 34.60.17235.2biotin-targeted
nanoparticle 37.10.2208.7tretinoin targeted
nanoparticle 370.2206dual targeted
nanoparticles
0
10
20
30
40
50
60
70
80
010 20 30 40 50 60 70 80
cumulated genistein release
time (hours)
GENSTEIN
TRETINOIN
No. Carriers Yield
Re
-
dispersible
index
F1 1- Manitol 5 % 78.78% 1.13
F2 2- Manitol : leucine :
formula (3 : 1) 78.9 % 0.91
F3 3- Manitol,
maltodextrin and
leucine (1.5 : 1.5 :1) 76.895% 1.071
F4 4- Hydroxyl propyl β-
cyclodextrin : manitol : leucine
(1.5 : 1.5 : 1 ) 71.8 % 0.90
Inhalable core-shell hybrid lipid-protein nanocomposites for tumor-targeted co-
delivery of genistein and tretinointo lung carcinoma
Nayra M. Kamel a,b, Maged W. Helmy c, Doaa M. Ragab a, Magda W. Samaha a,b, Ahmed O. Elzoghby a,b
a Department of Industrial Pharmacy, Faculty of Pharmacy, University of Alexandria, Alexandria, Egypt
b Cancer Nanotechnology Research Laboratory (CNRL), Faculty of Pharmacy, Alexandria University, Alexandria, Egypt
cDepartment of Pharmacology and Toxicology, Faculty of Pharmacy, Damanhur University, Elbehira, Egypt
0
5
10
15
20
25
30
35
40
45
Free ATRA Free GNS Free
combined
drugs
biotin
targeted
NP
tretinoin
targeted
NP
dual
targeted
NP
non
targeted
NP
IC50 values for different formulas
24 hr
48 hr