Improved Safety and Anti-Glioblastoma Efficacy of CAT3-Encapsulated SMEDDS through Metabolism Modification
Abstract
:1. Introduction
2. Results
2.1. Dispersibility Test, Droplet Size, PDI, and Zeta Potential
2.2. The Stability of CAT3-SMEDDS in Artificial Gastrointestinal Fluid
2.3. CAT3 Release Study
2.4. Transepithelial Transport of CAT3-SMEDDS
2.5. In Vitro Cellular Uptake of SMEDDS
2.6. In Vivo Plasma Pharmacokinetic and Brain Distribution Study
2.7. In Vivo Anti-Glioma Effect of CAT3-SMEDDS
2.8. Preliminary Safety Evaluation
3. Discussion
4. Materials and Methods
4.1. Materials
4.2. Cell Culture
4.3. Animals
4.4. Preparation and Characterization of CAT3-SMEDDS
4.5. Stability of CAT3-SMEDDS in Artificial Gastrointestinal Fluid
4.6. In Vitro Release
4.7. Bidirectional Transport of CAT3-SMEDDS in MDCK-MDR1 Monolayer
4.8. In Vitro Cellular Uptake of SMEDDS
4.9. In Vivo Pharmacokinetic Study in Plasma and Drug Distribution in Brain
4.10. Bioanalysis of CAT3 and PF403 in Rat Plasma and Brain Tissue Using LC-MS/MS
4.11. In Vivo Anti-Glioma Effect of CAT3-SMEDDS
4.12. Preliminary In Vivo Safety Evaluation
4.13. Statistical Analyses
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Sample Availability
Abbreviations
CAT3-SMEDDS | CAT3-loaded self-microemulsifying drug delivery system |
GBM | glioblastoma multiforme |
CNS | central nervous system |
TMZ | temozolomide |
SMEDDS | self-microemulsifying drug delivery system |
API | active pharmaceutical ingredient |
HPLC | high-performance liquid chromatography |
C6-luc | luciferase-expressing C6 cells |
DI | deionized |
PDI | polydispersity index |
SGF | simulated gastric fluid |
SIF | simulated intestinal fluid |
HBSS | Hanks’ balanced salt solution |
AP | apical side |
BL | basolateral |
LC-MS/MS | liquid chromatography-mass spectrometry |
Papp | apparent permeability coefficient |
PBS | phosphate-buffered saline |
Cou-6 | fluorescent Coumarin 6 |
ISTD | internal standard |
DAS | Drug and Statistics |
ER | efflux ratio |
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Name | Papp (×10−6 cm/s) | ER | |||
---|---|---|---|---|---|
AP-BL | % | BL-AP | % | ||
CAT3 | 0.1056 ± 0.0289 | 100 | 0.1021 ± 0.0154 | 100.0 | 0.9662 |
PF403 | 0.1617 ± 0.0345 | 153.2 | 0.4877 ± 0.1571 | 477.7 | 3.0155 |
CAT3-SMEDDS | 0.4129 ± 0.1103 | 391.0 | 0.2411 ± 0.1239 | 236.2 | 0.5840 |
Parameter | CAT3-API | CAT3-SMEDDS | CAT3-API+ Inhibition | SMEDDS+ Inhibition | |
---|---|---|---|---|---|
AUC(0–t) | ng/(mL·h) | 5.911 ± 0.396 | 10.564 ± 0.154 | 1.862 ± 0.356 | 1.235 ± 0.166 |
AUC(0–∞) | ng/(mL·h) | 7.369 ± 0.625 | 12.499 ± 1.928 | 1.971 ± 0.242 | 1.616 ± 0.645 |
MRT(0–t) | H | 4.208 ± 0.456 | 7.247 ± 0.08 | 7.369 ± 0.669 | 14.498 ± 0.429 |
MRT(0–∞) | H | 13.033 ± 6.792 | 12.18 ± 4.651 | 9.042 ± 1.923 | 24.766 ± 17.932 |
t1/2z | H | 15.472 ± 6.988 | 10.013 ± 4.243 | 5.364 ± 2.704 | 37.115± |
Tmax | H | 0.167 ± 0 | 0.167 ± 0 | 0.167 ± 0 | 0.083 ± 0 |
Vz/F | L/kg | 29,688.6957 ± 11,885.0772 | 11,261.752 ± 2910.449 | 41,374.9424 ± 26,075.1155 | 22916.9716± |
CLz/F | L/h/kg | 1363.91521 ± 121.36206 | 811.881 ± 115.054 | 5128.74435 ± 674.216985 | 6823.5499 ± 2437.732 |
Cmax | ng/mL | 5.811 ± 0.313 | 2.331 ± 0.022 | 0.361 ± 0.012 | 0.257 ± 0.043 |
Parameter | CAT3-API | CAT3-SMEDDS | CAT3-API+ Inhibition | SMEDDS+ Inhibition | |
---|---|---|---|---|---|
AUC(0–t) | ng/(L·h) | 20.537 ± 0.256 | 10.008 ± 0.677 | 5.747 ± 0.333 | 5.542 ± 0.455 |
AUC(0–∞) | ng/(L·h) | 26.578 ± 0.658 | 13.397 ± 6.082 | 7.018 ± 0.974 | 84.891 ± 81.228 |
MRT(0–t) | h | 9.73 ± 0.106 | 14.045 ± 0.42 | 9.998 ± 0.222 | 13.269 ± 0.828 |
MRT(0–∞) | h | 16.373 ± 1.002 | 22.828 ± 15.377 | 14.785 ± 2.65 | 403.858 ± 404.671 |
t1/2z | h | 10.348 ± 0.982 | 29.824 ± 1.996 | 8.602 ± 2.642 | 416.244 ± 218.378 |
Tmax | h | 8 ± 0 | 0.167 ± 0 | 8 ± 0 | 0.167 ± 0 |
Vz/F | L/kg | 5611.71866 ± 390.472773 | 21,117.8511 ± 1760.5231 | 17,410.146 ± 2991.1324 | 47,975.427 ± 1349.4173 |
CLz/F | L/h/kg | 376.406394 ± 9.18153 | 839.472493 ± 309.367699 | 1443.4282 ± 200.42835 | 614.21329 ± 905.19275 |
Cmax | ng/L | 1.574 ± 0.075 | 0.766 ± 0.036 | 0.548 ± 0.021 | 0.431 ± 0.05 |
Group | CAT3 in Plasma | Metabolite PF403 in Plasma | ||
---|---|---|---|---|
AUC(0–t) ng/L·h | Relative AUC(0–t)(%) | AUC(0-t) ng/L·h | Relative AUC(0–t)(%) | |
CAT3 | 5.911 ± 0.396 | 100.00 | 20.537 ± 0.256 | 100.00 |
SMEDDS | 10.564 ± 0.154 ** | 178.72 | 10.008 ± 0.677 ** | 48.73 |
CAT3 + Inhibition | 1.862 ± 0.356 ** | 31.50 | 5.747 ± 0.333 ** | 29.98 |
SMEDDS + Inhibition | 1.235 ± 0.166 ## | 11.69 Note 1 | 5.542 ± 0.455 ## Note 1 | 55.38 Note 1 |
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Wang, H.; Li, L.; Ye, J.; Dong, W.; Zhang, X.; Xu, Y.; Hu, J.; Wang, R.; Xia, X.; Yang, Y.; et al. Improved Safety and Anti-Glioblastoma Efficacy of CAT3-Encapsulated SMEDDS through Metabolism Modification. Molecules 2021, 26, 484. https://doi.org/10.3390/molecules26020484
Wang H, Li L, Ye J, Dong W, Zhang X, Xu Y, Hu J, Wang R, Xia X, Yang Y, et al. Improved Safety and Anti-Glioblastoma Efficacy of CAT3-Encapsulated SMEDDS through Metabolism Modification. Molecules. 2021; 26(2):484. https://doi.org/10.3390/molecules26020484
Chicago/Turabian StyleWang, Hongliang, Lin Li, Jun Ye, Wujun Dong, Xing Zhang, You Xu, Jinping Hu, Rubing Wang, Xuejun Xia, Yanfang Yang, and et al. 2021. "Improved Safety and Anti-Glioblastoma Efficacy of CAT3-Encapsulated SMEDDS through Metabolism Modification" Molecules 26, no. 2: 484. https://doi.org/10.3390/molecules26020484