Micelle-Forming Block Copolymers Tailored for Inhibition of P-gp-Mediated Multidrug Resistance: Structure to Activity Relationship
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemicals
2.2. Synthesis of Monomers and Initiator
2.3. Synthesis of Thiazolidine-2-Thione Functional CTA 2-Cyano-5-oxo-5-(2-thioxo-1,3-thiazolidin-3-yl)pentan-2-yl Ethyl Carbonotrithioate (CTA-TT)
2.4. Synthesis of Hydrophilic Polymer Blocks A1 and A2 Based on PHPMA Copolymers
2.5. Purification of Hydrophobic Polymer Block B Based on PPO before the Reaction
2.6. Synthesis of Polymer Carriers: Diblock and Triblock Micellar Copolymers
2.6.1. Boc-Protected Diblock Copolymer PHPMA-b-PPO P1 and P2
2.6.2. Diblock Copolymers P3 and P4, PHPMA-b-PPO, with Deprotected Hydrazide Groups in Distilled Water at Elevated Temperature and Pressure
2.6.3. Diblock Copolymer P5, PHPMA-b-PPO, with a Deprotected Hydrazide Group by the Mixture of TFA/TIS/Water
2.6.4. Triblock Copolymer P6, PHPMA-b-PPO-b-PHPMA
2.7. Loading of Free PPO into the Diblock Polymer Micelles
2.8. Time-Dependent Micellar Stability
2.9. Critical Micellar Concentration
2.10. Physico-Chemical Characterisation
2.11. Cell Lines
2.12. Calcein Efflux Assay
3. Results and Discussion
3.1. Synthesis of Hydrophilic Blocks A1, A2 and Unloaded Polymers P1–P6
3.2. Size of the Particles (Dh) and Long-Term Stability of Non-Loaded Polymers P1–P6
3.3. CMC of Non-Loaded Polymers P2–P6
3.4. Diblock Copolymer Micelles Loaded with PPO, Samples P7–P14
3.5. Biological Activity
3.5.1. The Influence of the Detailed Structure of the Copolymer Carrier on the P-gp Inhibitory Capacity
3.5.2. The Effect of PPO Loading to the Micelles on P-gp Inhibition
4. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Appendix A
References
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Sample Code | Mn.103 (g/mol) | Ð | Functionality of TT a | Content of NHNH2 (mol%) | Dh, 37 °C (nm) |
---|---|---|---|---|---|
A1 | 7.2 | 1.06 | 0.99 | 6.0 | 6.1 |
A2 | 5.0 | 1.02 | 0.81 | 4.7 | 4.5 |
Polymer | Sample Code 1 | Mn (SEC) 2 (g/mol) | Ð | % of PPO in Copolymer 3 | Mn (FFF) 4 (g/mol) | % of Micelles | Dh, 37 °C (nm) | CMC5 (g L−1) |
---|---|---|---|---|---|---|---|---|
P1 | DB-Boc | 13.20 | 1.05 | 39.7 | - | - | 26.2 | - |
P2 | DB-Boc-P | 12.90 | 1.18 | 36.6 | 9.63 × 105 | 88% | 23.8 | 0.0503 |
P3 | DB-H2O | 14.11 | 1.09 | 35.1 | 1.20 × 106 | 88% | 26.6 | 0.0347 |
P4 | DB-H2O-P | 13.29 | 1.22 | 36.2 | 6.97 × 105 | 76.5% | 25.4 | 0.0558 |
P5 | DB-TFA | 13.72 | 1.11 | 40.4 | 1.15 × 106 | 88% | 27.4 | 0.0459 |
P6 | TB-H2O-P | 13.40 | 1.10 | 35.9 | 3.70 × 105 | 45.3% | 22.3 | 0.1307 |
Sample Code | PPO/mg | Copolymer (P4)/mg | wt % of Loaded PPO * |
---|---|---|---|
P4 | 0 | 25 | 0 |
P7 | 1.35 | 25.55 | 5.02 |
P8 | 2.63 | 23.33 | 10.13 |
P9 | 4.51 | 25.54 | 15.01 |
P10 | 6.11 | 24.21 | 20.15 |
P11 | 8.31 | 24.75 | 25.14 |
P12 | 11.53 | 26.82 | 30.07 |
P13 | 13.52 | 24.97 | 35.13 |
P14 | 8.26 | 12.46 | 39.86 |
Sample Code | % of Total PPO in Copolymer 1 | Mn (FFF) 2 (g/mol) | % of Micelle | Dh, 37 °C (nm) | CMC 3 (g L−1) |
---|---|---|---|---|---|
P4 | 36.2 | 6.97 × 105 | 76.5% | 25.4 | 0.0558 |
P7 | 40.3 | - | - | 26.6 | 0.0585 |
P8 | 44.9 | 5.69 × 105 | 75.3% | 28.4 | 0.0524 |
P9 | 47.2 | 7.60 × 105 | 80.2% | 27.4 | 0.0339 |
P10 | 50.8 | 6.19 × 105 | 78.7% | 30.8 | 0.0506 |
P11 | 55.5 | - | - | 31.6 | 0.0443 |
P12 | 58.0 | 7.15 × 105 | 71.6% | 33.2 | 0.0438 |
P13 | 61.3 | - | - | 39.2 | 0.0422 |
P14 | 64.4 | 5.73 × 105 | 70.2% | 42.0 | - |
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Braunová, A.; Kaňa, M.; Kudláčová, J.; Kostka, L.; Bouček, J.; Betka, J.; Šírová, M.; Etrych, T. Micelle-Forming Block Copolymers Tailored for Inhibition of P-gp-Mediated Multidrug Resistance: Structure to Activity Relationship. Pharmaceutics 2019, 11, 579. https://doi.org/10.3390/pharmaceutics11110579
Braunová A, Kaňa M, Kudláčová J, Kostka L, Bouček J, Betka J, Šírová M, Etrych T. Micelle-Forming Block Copolymers Tailored for Inhibition of P-gp-Mediated Multidrug Resistance: Structure to Activity Relationship. Pharmaceutics. 2019; 11(11):579. https://doi.org/10.3390/pharmaceutics11110579
Chicago/Turabian StyleBraunová, Alena, Martin Kaňa, Júlia Kudláčová, Libor Kostka, Jan Bouček, Jan Betka, Milada Šírová, and Tomáš Etrych. 2019. "Micelle-Forming Block Copolymers Tailored for Inhibition of P-gp-Mediated Multidrug Resistance: Structure to Activity Relationship" Pharmaceutics 11, no. 11: 579. https://doi.org/10.3390/pharmaceutics11110579