Cetuximab Immunoliposomes Enhance Delivery of 5-FU to Skin Squamous Carcinoma Cells

Abstract

Topical chemotherapy of skin cancers is a promising strategy for reduction of side effects and for improvement of patient compliance. The combination of the chemotherapeutic 5-fluouracil (5-FU) and the anti-EGFR antibody cetuximab is a strategy to inhibit tumor growth. Their skin penetration, however, is hampered by their high hydrophilicity, which could be improved by encapsulation in delivery systems. Furthermore, it is a challenge to encapsulate hydrophilic drugs. The conjugation of an antibody to a liposome, maintaining its activity, is also a difficult task. Thus, we aimed to develop 5-FU liposomes and cetuximab-conjugated liposomes (immunoliposomes) of 5-FU to improve drug cytotoxicity against skin cancer cells. We characterized them by particle size, zeta potential, loading efficiency and antibody integrity. To optimize the loading efficiency of 5-FU, a series of liposomes were prepared, using different methods and drug-to-lipid ratios. Liposomes containing DSPC and Chol at drug-to-lipid ratio 0.1 prepared by the thin lipid hydration method resulted in the best 5-FU encapsulation and were chosen to conjugate with cetuximab. Cetuximab was directly coupled to preformed liposomes using DSPE-mPEG2000-Mal as an anchor. In A431 skin carcinoma cells, at 72 h, 5-FU liposomes showed a 5-fold lower IC50 than 5-FU solution. Additionally, 5-FU immunoliposomes resulted in a 4-fold lower cetuximab IC50 than cetuximab solution, demonstrating synergism with a combination index lower than 1 and potential to improve 5-FU and cetuximab cytotoxicity.

The Design of Anionic Surfactant-Based Amino-Functionalized Mesoporous Silica Nanoparticles and their Application in Transdermal Drug Delivery

“…However, DEX exhibited higher skin permeability in comparison to 5-FU [51]. The lower skin permeability of 5-FU compared to DEX could be attributed to the barrier properties of the stratum corneum and the aqueous solubility of 5-FU [52,53]. With MSN-NH 2 , a relative burst in the permeability of DEX and 5-FU was observed in the first 4 h, after which the rate of drug permeability was sustained to up to 48 h. Table 4 summarizes drug permeability parameters of the ex vivo diffusion study.…”

<p>Development, Characterization and Use of Liposomes as Amphipathic Transporters of Bioactive Compounds for Melanoma Treatment and Reduction of Skin Inflammation: A Review</p>

“…The solvent is evaporated by rotary evaporation and the dried lipid layer is then resuspended in PBS. 13 UDL has been prepared by reverse-phase evaporation, optimized by response surface methodology and loaded with papain to treat hypertrophic scar in New Zealand white rabbits. As result, the scar became thickened, collagen fibers had regular arrangement, and micro-vessels decreased, demonstrating that UDL is an efficient topical treatment against hypertrophic scars.…”

“…The development of low toxicity treatments, such as liposomes, has the advantage that the composition of these vesicles is closely similar to cell membranes, since they are elaborated with phospholipids and cholesterol. 13,76 There is valuable scientific information regarding the use of liposomes for skin delivery of therapeutic compounds for treatment of skin cancer or for enhancing liposome permeability to improve drug delivery ( Table 2). For example, Immunoliposomes loaded with 5-fluorouracil and cetuximab have been used against skin cancer cells A431.…”

“…Therefore, newly developed alternative treatments, such as the use of liposomes as drug carriers to protect against several skin pathologies need to be implemented. 13 Liposomes have been used in the cosmetic and drug industries because they can carry and protect encapsulated substances, improve the solubility of certain substances, and provide controlled drug release. 9 Some liposomes have been prepared for treatment of certain pathologies, such as melanoma, in which their use has a positive effect in reducing tumor size/weight and melanoma cell proliferation.…”

Carreador lipídico nanoestruturado multifuncional contendo 5-fluorouracil e siRNA para tratamento de câncer de pele

“…Diante desta toxicidade, o 5FU passou a ser amplamente utilizado associado a nanotecnologia com o intuito da diminuição da dose e dos efeitos adversos, melhor direcionamento e diminuição da degradação do fármaco. Vários tipos de nanopartículas já foram reportadas contendo 5FU para câncer, visando aplicações tópicas ou sistêmicas(CAMACHO et al, 2016;EL-HAMMADI et al, 2017;MIRANDA DA COSTA;MORAES, 2003;PETRILLI et al, 2017;SAGGIORO;SUBUDHI et al, 2015;TUMMALA;SATISH KUMAR;PRAKASH, 2015;…”