RESUMO
Combination chemotherapy must strike a difficult balance between safety and efficacy. Current regimens suffer from poor therapeutic impact because drugs are given at their maximum tolerated dose (MTD), which compounds the toxicity risk and exposes tumors to non-optimal drug ratios. A modular framework has been developed that selectively delivers drug combinations at synergistic ratios via tumor-targeting aptamers for effective low-dose treatment. A nucleolin-recognizing aptamer was coupled to peptide scaffolds laden with precise ratios of doxorubicin (DOX) and camptothecin (CPT). This construct had an extremely low IC50 (31.9â nm) against MDA-MB-231 breast cancer cells inâ vitro, and exhibited inâ vivo efficacy at micro-dose injections (500 and 350â µg kg-1 dose-1 of DOX and CPT, respectively) that are 20-30-fold lower than their previously-reported MTDs. This approach represents a generalizable strategy for the safe and consistent delivery of combination drugs in oncology.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Aptâmeros de Nucleotídeos/química , Camptotecina/uso terapêutico , Doxorrubicina/uso terapêutico , Neoplasias/tratamento farmacológico , Peptídeos/química , Protocolos de Quimioterapia Combinada Antineoplásica/química , Camptotecina/química , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Doxorrubicina/química , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Dose Máxima Tolerável , Estrutura Molecular , Neoplasias/patologiaRESUMO
Delivery of multiple therapeutics has become a preferred method of treating cancer, albeit differences in the biodistribution and pharmacokinetic profiles of individual drugs pose challenges in effectively delivering synergistic drug combinations to and at the tumor site. Here, bicompartmental Janus nanoparticles comprised of domains are reported with distinct bulk properties that allow for independent drug loading and release. Programmable drug release can be triggered by a change in the pH value and depends upon the bulk properties of the polymers used in the respective compartments, rather than the molecular structures of the active agents. Bicompartmental nanoparticles delivering a synergistic combination of lapatinib and paclitaxel result in increased activity against HER2+ breast cancer cells. Surprisingly, the dual drug loaded particles also show significant efficacy toward triple negative breast cancer, even though this cancer model is unresponsive to lapatinib alone. The broad versatility of the nanoparticle platform allows for rapid exploration of a wide range of drug combinations where both their relative drug ratios and temporal release profiles can be optimized.
Assuntos
Antineoplásicos , Nanopartículas , Neoplasias , Protocolos de Quimioterapia Combinada Antineoplásica , Linhagem Celular Tumoral , Combinação de Medicamentos , Sistemas de Liberação de Medicamentos , Humanos , Paclitaxel , Distribuição TecidualRESUMO
Soluto-inertial (SI) suspension interactions allow colloidal particles to be driven large distances over sustained periods of time. These interactions involve soluto-inertial "beacons" that establish and maintain solute fluxes over long times by slowly absorbing or emitting solutes in response to changes in the surrounding solution. Suspended particles then migrate in response to solute fluxes via diffusiophoresis (DP). Beacon materials must be chosen to maintain these solute fluxes, with range and duration in mind. Here we present a general strategy to facilitate qualitative design and quantitative prediction of SI interactions for a given beacon-solute pair. Specifically, we look at two classes of SI beacons: those that partition solute and those that associate with solute. We identify the design parameters for these systems to construct a parameter space map, calculate characteristic timescales over which SI fluxes persist, and generate approximate analytical expressions for solute concentration profiles. Further, we use these expressions to predict the DP velocity of colloids interacting with beacons, noting qualitative differences between beacon sources that release solute and beacon sinks that absorb solute. Proof-of-principle experiments of beacon sources and sinks, of partitioning, and associating types highlight the basic findings. More broadly, the conceptual approach outlined here can be adapted to treat SI interactions mediated by other materials such as dissolving solids, gases, evaporating liquids, ion-exchange resins, and others.
RESUMO
Combination chemotherapy is commonly used to treat late stage cancer; however, treatment is often limited by systemic toxicity. Optimizing drug ratio and schedule can improve drug combination activity and reduce dose to lower toxicity. Here, we identify gemcitabine (GEM) and doxorubicin (DOX) as a synergistic drug pair in vitro for the triple negative breast cancer cell line MDA-MB-231. Drug synergy and caspase activity were increased the most by exposing cells to GEM prior to DOX in vitro. While the combination was more effective than the single drugs at inhibiting MDA-MB-231 growth in vivo, the clear schedule dependence observed in vitro was not observed in vivo. Differences in drug exposure and cellular behavior in vivo compared to in vitro are likely responsible. This study emphasizes the importance in understanding how schedule impacts drug synergy and the need to develop more advanced strategies to translate synergy to the clinic.
RESUMO
Combination chemotherapy is commonly used to treat advanced breast cancer. However, treatment success is often limited due to systemic toxicity. To improve therapeutic efficacy, polymer drug conjugates carrying synergistic pairs of chemotherapy drugs can be used to reduce drug administration dose. Here, we systematically evaluated the effect of temporal scheduling of doxorubicin (DOX) and gemcitabine (GEM) on drug synergy. Hyaluronic acid (HA) drug conjugates with distinct linkers conjugating both DOX and GEM were synthesized to control relative release kinetics of each drug. We show that polymer conjugates that release GEM faster than DOX are more effective at killing triple negative breast cancer cells in vitro. We further show that the optimal dual drug conjugate more effectively inhibits the growth of an aggressive, orthotopic 4T1 tumor model in vivo than free DOX and GEM and the single drug HA conjugates. The dual drug HA conjugate can inhibit 4T1 tumor growth in vivo during treatment through both intravenous and non-local subcutaneous injections. These results emphasize the importance of understanding the effect release rates have on the efficacy of synergistic drug carriers and motivate the use of HA as a delivery platform for multiple cancer types.
Assuntos
Antibióticos Antineoplásicos/administração & dosagem , Antimetabólitos Antineoplásicos/administração & dosagem , Desoxicitidina/análogos & derivados , Doxorrubicina/administração & dosagem , Ácido Hialurônico/administração & dosagem , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Animais , Antibióticos Antineoplásicos/química , Antimetabólitos Antineoplásicos/química , Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Protocolos de Quimioterapia Combinada Antineoplásica/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Desoxicitidina/administração & dosagem , Desoxicitidina/química , Relação Dose-Resposta a Droga , Doxorrubicina/química , Portadores de Fármacos/administração & dosagem , Portadores de Fármacos/química , Liberação Controlada de Fármacos , Sinergismo Farmacológico , Humanos , Ácido Hialurônico/química , Camundongos , Camundongos Endogâmicos BALB C , Pró-Fármacos/administração & dosagem , Pró-Fármacos/química , GencitabinaRESUMO
PEGylated liposomes have transformed chemotherapeutic use of doxorubicin by reducing its cardiotoxicity; however, it remains unclear whether liposomal doxorubicin is therapeutically superior to free doxorubicin. Here, we demonstrate a novel PEGylated liposome system, named DAFODIL (Doxorubicin And 5-Flurouracil Optimally Delivered In a Liposome) that inarguably offers superior therapeutic efficacies compared to free drug administrations. Delivery of synergistic ratios of this drug pair led to greater than 90% reduction in tumor growth of murine 4T1 mammary carcinoma in vivo. By exploiting synergistic ratios, the effect was achieved at remarkably low doses, far below the maximum tolerable drug doses. Our approach re-invents the use of liposomes for multi-drug delivery by providing a chemotherapy vehicle which can both reduce toxicity and improve therapeutic efficacy. This methodology is made feasible by the extension of the ammonium-sulfate gradient encapsulation method to nucleobase analogues, a liposomal entrapment method once conceived useful only for anthracyclines. Therefore, our strategy can be utilized to efficiently evaluate various chemotherapy combinations in an effort to translate more effective combinations into the clinic.
Assuntos
Antibióticos Antineoplásicos/administração & dosagem , Antimetabólitos Antineoplásicos/administração & dosagem , Doxorrubicina/administração & dosagem , Fluoruracila/administração & dosagem , Neoplasias/tratamento farmacológico , Animais , Antibióticos Antineoplásicos/química , Antibióticos Antineoplásicos/farmacocinética , Antibióticos Antineoplásicos/uso terapêutico , Antimetabólitos Antineoplásicos/química , Antimetabólitos Antineoplásicos/farmacocinética , Antimetabólitos Antineoplásicos/uso terapêutico , Linhagem Celular , Linhagem Celular Tumoral , Doxorrubicina/química , Doxorrubicina/farmacocinética , Doxorrubicina/uso terapêutico , Combinação de Medicamentos , Feminino , Fluoruracila/química , Fluoruracila/farmacocinética , Fluoruracila/uso terapêutico , Humanos , Lipossomos , Camundongos Endogâmicos BALB C , Neoplasias/metabolismo , Neoplasias/patologia , Polietilenoglicóis/química , Triptofano/química , Carga Tumoral/efeitos dos fármacosRESUMO
Recent advancements in microfluidic technology have allowed for the generation and control of complex chemical gradients; however, few general techniques can measure these spatio-temporal concentration profiles without fluorescent labeling. Here we describe a Fabry-Perot interferometric technique, capable of measuring concentration profiles in situ, without any chemical label, by tracking Fringes of Equal Chromatic Order (FECO). The technique has a sensitivity of 10(-5) RIU, which can be used to track local solute changes of ~0.05% (w/w). The technique is spatially resolved (1 µm) and easily measures evolving concentration fields with ~20 Hz rate. Here, we demonstrate by measuring the binary diffusion coefficients of various solutes and solvents (and their concentration-dependence) in both free solution and in polyethylene glycol diacrylate (PEG-DA) hydrogels.
RESUMO
Combinations of topoisomerase inhibitors I and II have been found to synergistically inhibit cancer cell growth in vitro, yet clinical studies of these types of combinations have not progressed beyond phase II trials. The results of clinical combinations of topoisomerase (top) I and II inhibitors typically fall within one of two categories: little to no improvement in therapeutic efficacy, or augmented toxicity compared to the single drug counterparts. Hence, despite the promising activity of top I and II inhibitor combinations in vitro, their clinical applicability has not been realized. Here, we report the use of polymer-drug conjugates as a means to co-deliver synergistic doses of top I and II inhibitors camptothecin (CPT) and doxorubicin (DOX) to tumors in vivo in a 4T1 breast cancer model. At specific molar ratios, DOX and CPT were found to be among the most synergistic combinations reported to date, with combination indices between 0.01 and 0.1. The identified optimal ratios were controllably conjugated to hyaluronic acid, and elicited significant tumor reduction of murine 4T1 breast cancer model when administered intravenously. This study elucidates a method to identify synergistic drug combinations and translate them to in vivo by preserving the synergistic ratio via conjugation to a carrier polymer, thus opening a promising approach to translate drug combinations to clinically viable treatment regimens.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Animais , Antineoplásicos Fitogênicos/administração & dosagem , Antineoplásicos Fitogênicos/química , Apoptose/efeitos dos fármacos , Camptotecina/administração & dosagem , Camptotecina/química , Linhagem Celular Tumoral , Doxorrubicina/administração & dosagem , Doxorrubicina/química , Sinergismo Farmacológico , Feminino , Humanos , Ácido Hialurônico/administração & dosagem , Ácido Hialurônico/química , Camundongos Endogâmicos BALB C , Neoplasias/tratamento farmacológico , Neoplasias/patologia , Inibidores da Topoisomerase I/administração & dosagem , Inibidores da Topoisomerase I/química , Inibidores da Topoisomerase II/administração & dosagem , Inibidores da Topoisomerase II/química , Carga Tumoral/efeitos dos fármacosRESUMO
The impact of physical and chemical modifications of nanoparticles on their biological function has been systemically investigated and exploited to improve their circulation and targeting. However, the impact of nanoparticles' flexibility (i.e., elastic modulus) on their function has been explored to a far lesser extent, and the potential benefits of tuning nanoparticle elasticity are not clear. Here, we describe a method to synthesize polyethylene glycol (PEG)-based hydrogel nanoparticles of uniform size (200 nm) with elastic moduli ranging from 0.255 to 3000 kPa. These particles are used to investigate the role of particle elasticity on key functions including blood circulation time, biodistribution, antibody-mediated targeting, endocytosis, and phagocytosis. Our results demonstrate that softer nanoparticles (10 kPa) offer enhanced circulation and subsequently enhanced targeting compared to harder nanoparticles (3000 kPa) in vivo. Furthermore, in vitro experiments show that softer nanoparticles exhibit significantly reduced cellular uptake in immune cells (J774 macrophages), endothelial cells (bEnd.3), and cancer cells (4T1). Tuning nanoparticle elasticity potentially offers a method to improve the biological fate of nanoparticles by offering enhanced circulation, reduced immune system uptake, and improved targeting.
Assuntos
Sangue/metabolismo , Módulo de Elasticidade , Hidrogéis/química , Hidrogéis/metabolismo , Nanopartículas , Fagocitose , Animais , Linhagem Celular Tumoral , Humanos , Hidrogéis/farmacocinética , Camundongos , Tamanho da Partícula , Polietilenoglicóis/química , Relação Estrutura-Atividade , Distribuição TecidualRESUMO
Targeted delivery of therapeutic and imaging agents in the vascular compartment represents a significant hurdle in using nanomedicine for treating hemorrhage, thrombosis, and atherosclerosis. While several types of nanoparticles have been developed to meet this goal, their utility is limited by poor circulation, limited margination, and minimal targeting. Platelets have an innate ability to marginate to the vascular wall and specifically interact with vascular injury sites. These platelet functions are mediated by their shape, flexibility, and complex surface interactions. Inspired by this, we report the design and evaluation of nanoparticles that exhibit platelet-like functions including vascular injury site-directed margination, site-specific adhesion, and amplification of injury site-specific aggregation. Our nanoparticles mimic four key attributes of platelets, (i) discoidal morphology, (ii) mechanical flexibility, (iii) biophysically and biochemically mediated aggregation, and (iv) heteromultivalent presentation of ligands that mediate adhesion to both von Willebrand Factor and collagen, as well as specific clustering to activated platelets. Platelet-like nanoparticles (PLNs) exhibit enhanced surface-binding compared to spherical and rigid discoidal counterparts and site-selective adhesive and platelet-aggregatory properties under physiological flow conditions in vitro. In vivo studies in a mouse model demonstrated that PLNs accumulate at the wound site and induce â¼65% reduction in bleeding time, effectively mimicking and improving the hemostatic functions of natural platelets. We show that both the biochemical and biophysical design parameters of PLNs are essential in mimicking platelets and their hemostatic functions. PLNs offer a nanoscale technology that integrates platelet-mimetic biophysical and biochemical properties for potential applications in injectable synthetic hemostats and vascularly targeted payload delivery.
Assuntos
Plaquetas/patologia , Forma Celular , Nanopartículas , Lesões do Sistema Vascular/patologia , Animais , Adesão Celular , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Eletrônica de Varredura , Agregação Plaquetária , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
The impact of transport of surfactants to fluid-fluid interfaces is complex to assess and model, as many processes are in the regime where kinetics, diffusion and convection are comparable. Using the principle that the timescale for diffusion decreases with increasing curvature, we previously developed a microtensiometer to accurately measure fundamental transport coefficients via dynamic surface tension at spherical microscale liquid-fluid interfaces. In the present study, we use a low Reynolds number flow in the bulk solution to further increase the rate of diffusion. Dynamic surface tension is measured as a function of Peclet number and the results are compared with a simplified convection-diffusion model. Although a transition from diffusion to kinetic-limited transport is not observed experimentally for the surfactants considered, lower bounds on the adsorption and desorption rate constants are determined that are much larger than previously reported rate constants. The results show that the details of the flow field do not need to be controlled as long as the local Reynolds number is low. Aside from other pragmatic advantages, this experimental tool and analysis allows the governing mechanisms of surfactant transport at liquid-fluid interfaces to be quantified using flow near the interface to decrease the length scale for diffusion, separating the relevant timescales.