EphA2-Receptor Targeted PEGylated Nanoliposomes for the Treatment of BRAFV600E Mutated Parent- and Vemurafenib-Resistant Melanoma

Abstract

The clinical outcomes of malignant melanoma have improved with the introduction of mitogen-activated protein kinase kinase (MEK) inhibitors. However, off-target toxicities of the MEK inhibitor trametinib (TMB) often result in dose interruption and discontinuation of therapy. The purpose of this study was to anchor a physically stable EphrinA1-mimicking peptide known as YSA (YSAYPDSVPMMS) on TMB-loaded PEGylated nanoliposomes (YTPLs), and evaluate them in BRAFV600E-mutated parent cells (lines A375 and SK-MEL-28) and vemurafenib-resistant cells lines (A375R and SK-MEL-28R) in melanoma. TMB-loaded PEGylated liposomes (TPL) functionalized with nickel-chelated phospholipids were prepared using a modified hydration method. The hydrodynamic diameter and zeta potential values of optimized YTPL were 91.20 ± 12.16 nm and –0.92 ± 3.27 mV, respectively. The drug release study showed TPL did not leak or burst release in 24 h. The hemolysis observed was negligible at therapeutic concentrations of TMB. A differential scanning calorimetry (DSC) study confirmed that TMB was retained in a solubilized state within lipid bilayers. YTPL showed higher intracellular uptake in parental cell lines compared to vemurafenib-resistant cell lines. Western blot analysis and a cytotoxicity study with the EphA2 inhibitor confirmed a reduction in EphA2 expression in resistant cell lines. Thus, EphA2 receptor-targeted nanoliposomes can be useful for metastatic melanoma-specific delivery of TMB.

Aerosolized Nanoliposomal Carrier of Remdesivir: An Effective Alternative for COVID-19 Treatment In Vitro

“…As per Patel et al , high entrapment was possibly the result of rapid distribution of the drug within the inner hydrophobic lipid layer, thus restricting the ability of the drug to diffuse out in the aqueous phase [ 32 ]. Moreover, because of the presence of mannitol or osmotic pressure generation, more drug was favored in the inner lipid layer than the outer lipid layer during the hydration step, promoting enhanced stability [ 20 , 32 ]. Liposomal formulations with compositions comprising lung surfactants like DPPC could be potential carriers for pulmonary drug delivery owing to their high biodegradability, controllable surface charge, sustained release behavior and capability of encapsulating hydrophobic compounds like Rem [ 33 , 34 ].…”

“…For entrapment efficiency determination, AL-Rem was loaded in Amicon ultra centrifugal filters (50,000) and centrifuged at 1110 RCF for 10 min (Darmstadt, Germany). The decant was analyzed for free drug using HPLC analysis [ 20 ]. The encapsulation efficiency of Rem was expressed as the percentage of drug encapsulated and calculated using the following formula: …”

Targeting EphA2 in cancer

“…Trametinib (TMB) is a MAP/ERK kinase (MEK) inhibitor, but its offtarget toxicities frequently prompt dose interruption as well as treatment discontinuation [111]. Thus, YTPL, an ephrin A1-mimicking peptide (YSA; amino acid sequence: YSAYPDSVPMMS) with high stability [112], has been anchored on TMB-loaded PEGylated nanoliposomes [75]. YTPL was shown to display elevated cell internalization in comparison with non-targeted nanoliposomes (TPL) due to receptor-associated uptake.…”

Mechanisms of Acquired BRAF Inhibitor Resistance in Melanoma: A Systematic Review

“…As described above, considerable research is being undertaken to identify potential new combinations of treatments that may limit or prevent the development of BRAF inhibitor resistance, or overcome resistance once developed. In addition to this, investigations are underway to improve existing therapies, such as employing nanovehicle technology to enhance the safety or targeted delivery of drugs, which may enable the use of higher doses or more potent combinations of existing agents [ 284 , 285 ]. Preclinical studies have investigated the encapsulation of MEK inhibitors in pegylated nanoliposomes for oral administration [ 284 ], or the topical administration of BRAF inhibitor-loaded nanovehicles into melanoma lesions using a microneedling technique [ 285 ].…”