RESUMEN
The systemic delivery of drugs to the body via circulation after oral administration is a preferred method of drug administration during cancer treatment given its ease of implementation. However, the physicochemical properties of many current anticancer drugs limit their effectiveness when delivered by systemic routes. The use of nanoparticles (NPs) has emerged as an effective means of overcoming the inherent limitations of systemic drug delivery. We provide herein an overview of various NP formulations that facilitate improvements in the efficacy of various anticancer drugs compared with the free drug. This review will be useful to the reader who is interested in the role NP technology is playing in shaping the future of chemotherapeutic drug delivery and disease treatment.
Asunto(s)
Antineoplásicos/administración & dosificación , Sistemas de Liberación de Medicamentos/métodos , Nanopartículas , Neoplasias/tratamiento farmacológico , Nanomedicina Teranóstica/métodos , Administración Oral , Animales , Modelos Animales de Enfermedad , Composición de Medicamentos/métodos , Humanos , Resultado del TratamientoRESUMEN
In this study, we developed a peptide-dendrimer-drug conjugate system for the pH-triggered direct cytosolic delivery of the cancer chemotherapeutic doxorubicin (DOX) using the pH Low Insertion Peptide (pHLIP). We synthesized a pHLIP-dendrimer-DOX conjugate in which a single copy of pHLIP displayed a generation three dendrimer bearing multiple copies of DOX via disulfide linkages. Biophysical analysis showed that both the dendrimer and a single DOX conjugate inserted into membrane bilayers in a pH-dependent manner. Time-resolved confocal microscopy indicate the single DOX conjugate may undergo a faster rate of membrane translocation, due to greater nuclear localization of DOX at 24â¯h and 48â¯h post delivery. At 72â¯h, however, the levels of DOX nuclear accumulation for both constructs were identical. Cytotoxicity assays revealed that both constructs mediated â¼80% inhibition of cellular proliferation at 10⯵M, the dendrimer complex exhibited a 17% greater cytotoxic effect at lower concentrations and greater than three-fold improvement in IC50 over free DOX. Our findings show proof of concept that the dendrimeric display of DOX on the pHLIP carrier (1) facilitates the pH-dependent and temporally-controlled release of DOX to the cytosol, (2) eliminates the endosomal sequestration of the drug cargo, and (3) augments DOX cytotoxicity relative to the free drug.
Asunto(s)
Antibióticos Antineoplásicos/metabolismo , Dendrímeros/química , Disulfuros/química , Doxorrubicina/metabolismo , Portadores de Fármacos , Endocitosis , Proteínas de la Membrana/química , Neoplasias del Cuello Uterino/metabolismo , Antibióticos Antineoplásicos/administración & dosificación , Antibióticos Antineoplásicos/química , Proliferación Celular/efectos de los fármacos , Preparaciones de Acción Retardada , Relación Dosis-Respuesta a Droga , Doxorrubicina/administración & dosificación , Doxorrubicina/química , Composición de Medicamentos , Femenino , Células HeLa , Humanos , Concentración de Iones de Hidrógeno , Concentración 50 Inhibidora , Cinética , Membranas Artificiales , Microscopía Confocal , Oxidación-Reducción , Tecnología Farmacéutica/métodos , Neoplasias del Cuello Uterino/tratamiento farmacológico , Neoplasias del Cuello Uterino/patologíaRESUMEN
The targeting of therapeutics specifically to diseased tissue is crucial for the development of successful cancer treatments. The approach here is based on the pH(low) insertion peptide (pHLIP) for the delivery of a potent mitotic inhibitor monomethyl auristatin F (MMAF). We investigated six pHLIP variants conjugated to MMAF to compare their efficacy in vitro against cultured cancer cells. While all pHLIP-MMAF conjugates exhibit potent pH- and concentration-dependent killing, their cytotoxicity profiles are remarkably different. We also show that the lead conjugate exhibits significant therapeutic efficacy in mouse models without overt toxicities. This study confirms pHLIP-monomethyl auristatin conjugates as possible new therapeutic options for cancer treatment and supports their further development.
Asunto(s)
Aminobenzoatos/farmacología , Antineoplásicos/farmacología , Inmunoconjugados/farmacología , Oligopéptidos/farmacología , Péptidos/farmacología , Animales , Anticuerpos Monoclonales/farmacología , Línea Celular , Línea Celular Tumoral , Femenino , Células HeLa , Humanos , Concentración de Iones de Hidrógeno , Ratones , Ratones DesnudosRESUMEN
Positively charged antimicrobial peptides have become promising agents for the treatment of cancer by inducing apoptosis though their preferential binding and disruption of negatively charged membranes, such as the mitochondrial membrane. (KLAKLAK)2 is such a peptide but due to its polarity, it cannot cross the cellular membrane and therefore relies on the use of a delivery agent. For targeted delivery, previous studies have relied on cell penetrating peptides, nanoparticles or specific biomarkers. Herein, we investigated the first use of pHLIP to selectively target and directly translocate (KLAKLAK)2 into the cytoplasm of breast cancer cells, based on the acidic tumor micro-environment. With the goal of identifying a lead conjugate with optimized selective cytotoxicity towards cancer cells, we analyzed a family of (KLAKLAK)2 analogs with varying size, polarity and charge. We present a highly efficacious pHLIP conjugate that selectively induces concentration- and pH-dependent toxicity in breast cancer cells.
Asunto(s)
Péptidos Catiónicos Antimicrobianos/química , Proteínas de la Membrana/química , Secuencia de Aminoácidos , Péptidos Catiónicos Antimicrobianos/síntesis química , Péptidos Catiónicos Antimicrobianos/toxicidad , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Portadores de Fármacos/química , Humanos , Concentración de Iones de Hidrógeno , Proteínas de la Membrana/síntesis química , Membranas Mitocondriales/efectos de los fármacos , Membranas Mitocondriales/metabolismo , Técnicas de Síntesis en Fase SólidaRESUMEN
Even though abnormal expression of G protein-coupled receptors (GPCRs) and of their ligands is observed in many cancer cells of various origins, only a few anti-cancer compounds directly act on their signalling. One promising approach to modulate their activity consists of targeting the receptor cytoplasmic surfaces interacting with the associated G-proteins using peptides mimicking the intracellular loops of the receptor. Thus, to be fully effective, the peptide mimics must be selectively targeted to the tumour while sparing healthy tissues, translocated across the cell membrane and stay anchored to the cytoplasmic leaflet of the plasma membrane. In the present study, we introduce a novel way to selectively target and inhibit the activity of a GPCR in cancer cells under acidic conditions, such as those found in solid tumours. We find that the conjugation of a peptide fragment derived from the third intracellular loop (i3) of the protease-activated receptor 1 (PAR1) to a peptide that can selectively target tumours solely based on their acidity [pH(Low) Insertion Peptide (pHLIP)], produces a construct capable of effectively down-regulating PAR1 activity in a concentration- and pH-dependent manner and of inducing a potent cytotoxic effect in a panel of cancer cells that is proportional to the relative level of receptor expression at the cell surface. This strategy not only allows for a more selective targeting and specific intracellular delivery than current approaches, but also offers new possibilities for developing novel anti-cancer drugs targeting GPCRs.
Asunto(s)
Regulación hacia Abajo/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Proteínas de la Membrana/farmacología , Proteínas de Neoplasias/biosíntesis , Neoplasias/tratamiento farmacológico , Receptor PAR-1/biosíntesis , Muerte Celular/efectos de los fármacos , Células HeLa , Humanos , Concentración de Iones de Hidrógeno , Células MCF-7 , Neoplasias/metabolismo , Neoplasias/patologíaRESUMEN
Localized delivery is vital for the successful development of novel and effective therapeutics for the treatment of cancer. The targeting and delivery described herein is based on the pH (low) insertion peptide (pHLIP), a unique delivery peptide that can selectively target tumors in mice and translocate and release cargo molecules intracellularly based solely on the low extracellular pH intrinsic to cancer cells. In this study, we investigate the efficacy of pHLIP to target and deliver the highly potent and clinically validated microtubule inhibitor monomethyl auristatin E (MMAE) to cancer cells and breast tumors. We show that pHLIP-MMAE conjugates induce a potent cytotoxic effect (>90% inhibition of cell growth) in a concentration- and pH-dependent manner after only 2 h incubation without any apparent disruption of the plasma membrane. pHLIP-MMAE conjugates exhibit between an 11- and 144-fold higher antiproliferative effect at low pH than that at physiological pH and a pronounced pH-dependent cytotoxicity as compared to that of free drug. Furthermore, we demonstrate that a pHLIP-MMAE drug conjugate effectively targets triple-negative breast tumor xenografts in mice. These results indicate that pHLIP-based auristatin conjugates may have an enhanced therapeutic window as compared to that of free drug, providing a targeting mechanism to attenuate systemic toxicity.