N-(2-Hydroxypropyl)methacrylamide copolymer conjugates of pirarubicin (THP), P-THP, accumulates selectively in solid tumor tissue by the enhanced permeability and retention (EPR) effect. Despite of high accumulation in solid tumors, some macromolecular antitumor agents show poor therapeutic outcome because of poor tissue diffusion into the tumor as well as obstructed tumor blood flow. Here, we confirmed that cellular uptake of P-THP was 25 times less than that of free THP at 1-4 h incubation time in vitro. The passage of P-THP through the confluent tight-monolayer cells junction was 12 times higher than free THP, and P-THP penetrated deeper into the tumor cell spheroid (1.3-1.7-fold) than free THP in 4 h. In addition, P-THP showed cytotoxicity comparable to that of free THP to tumor-cells in spheroid form, despite of 7 times lower cytotoxicity of P-THP to the monolayer cells to that of free THP in vitro. These results indicate that P-THP administration can exhibit deeper diffusion into the tumor cell spheroid than free THP. As a consequence, P-THP exhibits more efficient antitumor activity than free THP in vivo, which is also supported by better pharmacokinetics and tumor accumulation of P-THP than free THP.
- Klíčová slova
- HPMA polymer, drug delivery, penetration, pirarubicin (THP), tumor spheroid cell,
- MeSH
- akrylamidy chemie MeSH
- antitumorózní látky aplikace a dávkování farmakokinetika MeSH
- buněčné sféroidy MeSH
- doxorubicin aplikace a dávkování analogy a deriváty farmakokinetika MeSH
- HCT116 buňky MeSH
- léky antitumorózní - screeningové testy MeSH
- lidé MeSH
- nádory farmakoterapie patologie MeSH
- nosiče léků chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- akrylamidy MeSH
- antitumorózní látky MeSH
- doxorubicin MeSH
- N-(2-hydroxypropyl)methacrylamide MeSH Prohlížeč
- nosiče léků MeSH
- pirarubicin MeSH Prohlížeč
Ritonavir (RIT) is a widely used antiviral drug that acts as an HIV protease inhibitor with emerging potential in anticancer therapies. RIT causes inhibition of P-glycoprotein, which plays an important role in multidrug resistance (MDR) in cancer cells when overexpressed. Moreover, RIT causes mitochondrial dysfunction, leading to decreased ATP production and reduction of caveolin I expression, which can affect cell migration and tumor progression. To increase its direct antitumor activity, decrease severe side effects induced by the use of free RIT and improve its pharmacokinetics, ritonavir 5-methyl-4-oxohexanoate (RTV) was synthesized and conjugated to a tumor-targeted polymer carrier based on a N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer. Here we demonstrated that polymer-bound RTV enhanced the internalization of polymer-RTV conjugates, differing in RTV content from 4 to 15 wt%, in HeLa cancer cells compared with polymer without RTV. The most efficient influx and internalization properties were determined for the polymer conjugate bearing 11 wt% of RTV. This conjugate was internalized by cells using both caveolin- and clathrin-dependent endocytic pathways in contrast to the RTV-free polymer, which was preferentially internalized only by clathrin-mediated endocytosis. Moreover, we found the co-localization of the RTV-conjugate with mitochondria and a significant decrease of ATP production in treated cells. Thus, the impact on mitochondrial mechanism can influence the function of ATP-dependent P-glycoprotein and also the cell viability of MDR cancer cells. Overall, this study demonstrated that the polymer-RTV conjugate is a promising polymer-based nanotherapeutic, suitable for antitumor combination therapy with other anticancer drugs and a potential mitochondrial drug delivery system.
- Klíčová slova
- Anticancer therapy, Cell penetration compound, Drug delivery, HPMA, Mitochondrial drug delivery, Multidrug resistance, Polymer-based nanotherapeutic, Ritonavir,
- MeSH
- adenosintrifosfát biosyntéza MeSH
- antitumorózní látky aplikace a dávkování chemie MeSH
- chemorezistence účinky léků MeSH
- endocytóza účinky léků MeSH
- HeLa buňky MeSH
- kaveolin 1 biosyntéza genetika MeSH
- klathrin farmakologie MeSH
- koncentrace vodíkových iontů MeSH
- lidé MeSH
- methakryláty chemie MeSH
- nanostruktury chemie MeSH
- P-glykoprotein účinky léků metabolismus MeSH
- polymery MeSH
- ritonavir aplikace a dávkování analogy a deriváty chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- adenosintrifosfát MeSH
- antitumorózní látky MeSH
- hydroxypropyl methacrylate MeSH Prohlížeč
- kaveolin 1 MeSH
- klathrin MeSH
- methakryláty MeSH
- P-glykoprotein MeSH
- polymery MeSH
- ritonavir MeSH
Over the past few decades, numerous polymer drug carrier systems are designed and synthesized, and their properties are evaluated. Many of these systems are based on water-soluble polymer carriers of low-molecular-weight drugs and compounds, e.g., cytostatic agents, anti-inflammatory drugs, or multidrug resistance inhibitors, all covalently bound to a carrier by a biodegradable spacer that enables controlled release of the active molecule to achieve the desired pharmacological effect. Among others, the synthetic polymer carriers based on N-(2-hydroxypropyl) methacrylamide (HPMA) copolymers are some of the most promising carriers for this purpose. This review focuses on advances in the development of HPMA copolymer carriers and their conjugates with anticancer drugs, with triggered drug activation in tumor tissue and especially in tumor cells. Specifically, this review highlights the improvements in polymer drug carrier design with respect to the structure of a spacer to influence controlled drug release and activation, and its impact on the drug pharmacokinetics, enhanced tumor uptake, cellular trafficking, and in vivo antitumor activity.
- Klíčová slova
- HPMA copolymers, biodegradable spacer, controlled drug release, drug delivery systems, pH-controlled release,
- MeSH
- léky s prodlouženým účinkem MeSH
- lidé MeSH
- methakryláty chemie terapeutické užití MeSH
- nádory farmakoterapie MeSH
- nosiče léků chemie terapeutické užití MeSH
- uvolňování léčiv * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- hydroxypropyl methacrylate MeSH Prohlížeč
- léky s prodlouženým účinkem MeSH
- methakryláty MeSH
- nosiče léků MeSH
Polymer prodrugs can considerably improve the treatment of tumors with multidrug resistance, often caused by overexpression of P-glycoprotein (P-gp). Here, we present the effect of the N-(2-hydroxypropyl) methacrylamide-based polymer conjugate with P-gp inhibitor ritonavir (RIT) on the increase of free doxorubicin (DOX) and polymer-bound DOX cytotoxicity in the human neuroblastoma 4 cell line and its resistant clones to different cytostatics. The increase in cytotoxicity after polymer-RIT conjugate pretreatment was higher for the lines overexpressing P-gp and less pronounced for those with decreased P-gp levels. Moreover, the effect of polymer conjugate containing inhibitor and DOX on the same polymer chain was lower than that of two individual polymer conjugates used sequentially. In conclusion, the polymer-RIT conjugate can significantly increase the cytotoxicity of free DOX and polymer-DOX conjugates in cells with various multidrug resistance origins and can thus be considered a suitable therapeutic enhancer of polymer prodrugs.
- MeSH
- akrylamidy aplikace a dávkování farmakologie MeSH
- chemorezistence MeSH
- doxorubicin aplikace a dávkování farmakologie MeSH
- léky antitumorózní - screeningové testy MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- neuroblastom farmakoterapie metabolismus MeSH
- P-glykoprotein antagonisté a inhibitory biosyntéza MeSH
- protokoly antitumorózní kombinované chemoterapie farmakologie MeSH
- ritonavir aplikace a dávkování farmakologie MeSH
- synergismus léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- akrylamidy MeSH
- doxorubicin MeSH
- N-(2-hydroxypropyl)methacrylamide MeSH Prohlížeč
- P-glykoprotein MeSH
- ritonavir MeSH
Here, we describe a novel polymer platform suitable for efficient diagnostics and potential theranostics based on 89Zr-labeled N-(2-hydroxypropyl)methacrylamide (HPMA)-based copolymer conjugates. A set of polymers differing in molecular weight with either low dispersity or high dispersity were designed and synthesized and their biodistribution in vivo was successfully and precisely observed over 72 h. Moreover, the feasibility of two imaging techniques, fluorescence imaging (FI) and positron emission tomography (PET), was compared using labeled polymer conjugates. Both methods gave comparable results thus showing the enhanced diagnostic potential of the prepared polymer-dye or polymer-chelator-89Zr constructs. The in vivo and ex vivo PET/FI studies indicated that the dispersity and molecular weight of the linear HPMA polymers have a significant influence on the pharmacokinetics of the polymer conjugates. The higher molecular weight and narrower distribution of molecular weights of the polymer carriers improve their pharmacokinetic profile for highly prolonged blood circulation and enhanced tumor uptake. Moreover, the same polymer carrier with the anticancer drug doxorubicin bound by a pH-sensitive hydrazone bond showed higher cytotoxicity and cellular uptake in vitro. Therefore, HPMA copolymers with low dispersity and a molecular weight near the limit of renal filtration can be used as highly efficient polymer carriers of tumor-targeted therapeutics or for theranostics with minimal side effects.
- MeSH
- doxorubicin aplikace a dávkování farmakokinetika MeSH
- experimentální nádory diagnostické zobrazování farmakoterapie MeSH
- Jurkat buňky MeSH
- lidé MeSH
- MFC-7 buňky MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- nosiče léků chemie MeSH
- optické zobrazování * MeSH
- polymery chemie MeSH
- pozitronová emisní tomografie * MeSH
- radionuklidy MeSH
- teranostická nanomedicína * MeSH
- tkáňová distribuce MeSH
- zirkonium MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- doxorubicin MeSH
- nosiče léků MeSH
- polymery MeSH
- radionuklidy MeSH
- Zirconium-89 MeSH Prohlížeč
- zirkonium MeSH
Many conjugates of water-soluble polymers with biologically active molecules were developed during the last two decades. Although, therapeutic effects of these conjugates are affected by the properties of carriers, the properties of the attached drugs appear more important than the same carrier polymer in this case. Pirarubicin (THP), a tetrahydropyranyl derivative of doxorubicin (DOX), demonstrated more rapid cellular internalization and potent cytotoxicity than DOX. Here, we conjugated the THP or DOX to N-(2-hydroxypropyl)methacrylamide copolymer via a hydrazone bond. The polymeric prodrug conjugates, P-THP and P-DOX, respectively, had comparable hydrodynamic sizes and drug loading. Compared with P-DOX, P-THP showed approximately 10 times greater cellular uptake during a 240 min incubation and a cytotoxicity that was more than 10 times higher during a 72-h incubation. A marginal difference was seen in P-THP and P-DOX accumulation in the liver and kidney at 6 h after drug administration, but no significant difference occurred in the tumor drug concentration during 6-24 h after drug administration. Antitumor activity against xenograft human pancreatic tumor (SUIT2) in mice was greater for P-THP than for P-DOX. To sum up, the present study compared the biological behavior of two different drugs, each attached to an N-(2-hydroxypropyl)methacrylamide copolymer carrier, with regard to their uptake by tumor cells, body distribution, accumulation in tumors, cytotoxicity, and antitumor activity in vitro and in vivo. No differences in the tumor cell uptake of the polymer-drug conjugates, P-THP and P-DOX, were observed. In contrast, the intracellular uptake of free THP liberated from the P-THP was 25-30 times higher than that of DOX liberated from P-DOX. This finding indicates that proper selection of the carrier, and especially conjugated active pharmaceutical ingredient (API) are most critical for anticancer activity of the polymer-drug conjugates. THP, in this respect, was found to be a more preferable API for polymer conjugation than DOX. Hence the treatment based on enhanced permeability and retention (EPR) effect that targets more selectively to solid tumors can be best achieved with THP, although both polymer conjugates of DOX and THP exhibited the EPR effects and drug release profiles in acidic pH similarly.
- Klíčová slova
- EPR effect, HPMA polymer conjugate, acid-cleavable linkage, doxorubicin (DOX), pirarubicin (THP),
- MeSH
- akrylamidy chemie MeSH
- antibiotika antitumorózní chemie farmakologie MeSH
- antitumorózní látky chemie farmakologie MeSH
- doxorubicin analogy a deriváty chemie farmakologie MeSH
- experimentální sarkom farmakoterapie metabolismus patologie MeSH
- lidé MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- nádorové buňky kultivované MeSH
- nosiče léků aplikace a dávkování chemie MeSH
- polymery aplikace a dávkování chemie MeSH
- proliferace buněk účinky léků MeSH
- xenogenní modely - testy antitumorózní aktivity MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Názvy látek
- akrylamidy MeSH
- antibiotika antitumorózní MeSH
- antitumorózní látky MeSH
- doxorubicin MeSH
- N-(2-hydroxypropyl)methacrylamide MeSH Prohlížeč
- nosiče léků MeSH
- pirarubicin MeSH Prohlížeč
- polymery MeSH
Water-soluble N-(2-hydroxypropyl)methacrylamide copolymer conjugates bearing the anticancer drugs doxorubicin (Dox) or pirarubicin (THP), P-gp inhibitors derived from reversin 121 (REV) or ritonavir (RIT)), or both anticancer drug and P-gp inhibitor were designed and synthesized. All biologically active molecules were attached to the polymer carrier via pH-sensitive spacer enabling controlled release in mild acidic environment modeling endosomes and lysosomes of tumor cells. The cytotoxicity of the conjugates against three sensitive and Dox-resistant neuroblastoma (NB) cell lines, applied alone or in combination, was studied in vitro. All conjugates containing THP displayed higher cytotoxicity against all three Dox-resistant NB cell lines compared with the corresponding Dox-containing conjugates. Furthermore, the cytotoxicity of conjugates containing both drug and P-gp inhibitor was up to 10 times higher than that of the conjugate containing only drug. In general, the polymer-drug conjugates showed higher cytotoxicity when conjugates containing inhibitors were added 8 or 16h prior to treatment compared with conjugates bearing both the inhibitor and the drug. The difference in cytotoxicity was more pronounced at the 16-h time point. Moreover, higher inhibitor:drug ratios resulted in higher cytotoxicity. The cytotoxicity of the polymer-drug used in combination with polymer P-gp inhibitor was up to 84 times higher than that of the polymer-drug alone.
- Klíčová slova
- Doxorubicin, Multidrug resistance, N-(2-hydroxypropyl)methacrylamide copolymers, Neuroblastoma, P-glycoprotein inhibitors, Pirarubicin, Reversin 121, Ritonavir,
- MeSH
- antibiotika antitumorózní aplikace a dávkování chemie farmakologie MeSH
- chemorezistence účinky léků MeSH
- doxorubicin aplikace a dávkování analogy a deriváty chemie farmakologie MeSH
- lidé MeSH
- methakryláty aplikace a dávkování chemie MeSH
- mnohočetná léková rezistence účinky léků MeSH
- nádorové buněčné linie MeSH
- neuroblastom genetika metabolismus MeSH
- oligopeptidy aplikace a dávkování chemie farmakologie MeSH
- P-glykoprotein antagonisté a inhibitory genetika metabolismus MeSH
- ritonavir aplikace a dávkování chemie farmakologie MeSH
- uvolňování léčiv MeSH
- viabilita buněk účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antibiotika antitumorózní MeSH
- doxorubicin MeSH
- hydroxypropyl methacrylate MeSH Prohlížeč
- methakryláty MeSH
- oligopeptidy MeSH
- P-glykoprotein MeSH
- pirarubicin MeSH Prohlížeč
- reversin 121 MeSH Prohlížeč
- ritonavir MeSH
- Publikační typ
- časopisecké články MeSH
High-molecular-weight star polymer drug nanocarriers intended for the treatment and/or visualisation of solid tumours were synthesised, and their physico-chemical and preliminary in vitro biological properties were determined. The water-soluble star polymer carriers were prepared by the grafting of poly(amido amine) (PAMAM) dendrimers by hetero-telechelic N-(2-hydroxypropyl)methacrylamide (HPMA) copolymers, synthesised by the controlled radical Reversible Addition Fragmentation chain Transfer (RAFT) polymerisation. The well-defined star copolymers with Mw values ranging from 2 · 10(5) to 6 · 10(5) showing a low dispersity (approximately 1.2) were prepared in a high yield. A model anticancer drug, doxorubicin, was bound to the star polymer through a hydrazone bond, enabling the pH-controlled drug release in the target tumour tissue. The activated polymer arm ends of the star copolymer carrier enable a one-point attachment for the targeting ligands and/or a labelling moiety. In this study, the model TAMRA fluorescent dye was used to prove the feasibility of the polymer carrier visualisation by optical imaging in vitro. The tailor-made structure of the star polymer carriers should facilitate the synthesis of targeted polymer-drug conjugates, even polymer theranostics, for simultaneous tumour drug delivery and imaging.
- Klíčová slova
- HPMA copolymers, drug delivery systems, pH-controlled release, reversible addition fragmentation chain transfer (RAFT), star copolymers,
- MeSH
- dendrimery * chemická syntéza chemie farmakokinetika farmakologie MeSH
- doxorubicin * chemie farmakokinetika farmakologie MeSH
- léky s prodlouženým účinkem chemická syntéza chemie farmakokinetika farmakologie MeSH
- lidé MeSH
- methakryláty * chemie farmakokinetika farmakologie MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- nádory * farmakoterapie metabolismus patologie MeSH
- nanočástice chemie ultrastruktura MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- dendrimery * MeSH
- doxorubicin * MeSH
- hydroxypropyl methacrylate MeSH Prohlížeč
- léky s prodlouženým účinkem MeSH
- methakryláty * MeSH
- PAMAM Starburst MeSH Prohlížeč
Previously we showed that linear poly(N-(2-hydroxypropyl)methacrylamide) conjugates of pirarubicin (THP), LP-THP, with MW about 39 kDa, exhibited far better tumor accumulation and therapeutic effect than that of parental free THP. To improve the pharmacokinetics of LP-THP further, high-MW conjugate of poly(amido amine) (PAMAM) dendrimer grafted with semitelechelic HPMA copolymer (PHPMA) was synthesized [star polymer (SP); 400 kDa] and conjugated with THP via hydrazone bond-containing spacer (SP-THP). THP was conjugated to SP to form SP-THP via acid cleavable hydrazone bonding, which responds to acidic milieu of tumor tissue. As a consequence, it would release free THP, by active therapeutic principle. SP-THP exhibits larger hydrodynamic diameter (25.9 nm) in aqueous solution than that of LP-THP (8.2 nm) as observed by light scattering and size exclusion chromatography. Because of the larger size, the tumor AUC5h-72 h of SP-THP was 3.3 times higher than that of LP-THP. More importantly, released free THP was retained selectively in the tumor tissue for at least up to 72 h after administration of SP-THP. We found that SP-THP exhibited superior antitumor effect to LP-THP against both S-180 tumor-bearing mice in vivo, and with chemically AOM/DSS-induced colon tumor-bearing mice, most probably due to their different molecular size. In our comparison study of in vitro and in vivo behavior of SP-THP and LP-THP we concluded that SP-THP exhibited enhanced therapeutic efficacy not only in implanted tumor but also in orthotopic/spontaneous tumor despite its higher toxicity compared to LP-THP. Upon these findings further investigation using various tumors including transgenic, and metastatic tumors is going to be conducted soon.
- Klíčová slova
- Acid-cleavable linkage, Chemical carcinogenesis, Controlled drug release, Dendrimer-derived polymer conjugate, EPR effect, HPMA polymer conjugate, Pirarubicin (THP),
- MeSH
- antitumorózní látky chemie farmakokinetika farmakologie MeSH
- berberinové alkaloidy chemie farmakokinetika MeSH
- dendrimery chemie farmakokinetika MeSH
- doxorubicin analogy a deriváty chemie farmakokinetika farmakologie MeSH
- HeLa buňky MeSH
- lidé MeSH
- melanom experimentální MeSH
- methakryláty chemie farmakokinetika MeSH
- myši inbrední ICR MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- nádory farmakoterapie MeSH
- nosiče léků chemie farmakokinetika MeSH
- polymery chemie farmakokinetika MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- 2,3,10,11-tetrahydroxytetrahydroprotoberberine MeSH Prohlížeč
- antitumorózní látky MeSH
- berberinové alkaloidy MeSH
- dendrimery MeSH
- doxorubicin MeSH
- hydroxypropyl methacrylate MeSH Prohlížeč
- methakryláty MeSH
- nosiče léků MeSH
- PAMAM Starburst MeSH Prohlížeč
- pirarubicin MeSH Prohlížeč
- polymery MeSH