Gemini Lipopeptide Bearing an Ultrashort Peptide for Enhanced Transfection Efficiency and Cancer-Cell-Specific Cytotoxicity

Abstract

Cationic gemini lipopeptides are a relatively new class of amphiphilic compounds to be used for gene delivery. Through the possibility of incorporating short peptides with cell-penetrating functionalities, these lipopeptides may be advantageous over traditional cationic lipids. Herein, we report the design, synthesis, and application of a novel cationic gemini lipopeptide for gene delivery. An ultrashort peptide, containing four amino acids, arginine–cysteine–cysteine–arginine, serves as a cationic head group, and two α-tocopherol moieties act as hydrophobic anchoring groups. The new lipopeptide (ATTA) is incorporated into the conventional liposomes, containing 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) and 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine (DOPE), at different molar ratios. The formulated liposomes are characterized and screened for better transfection efficiency. Transfection activity in multiple human cell lines from cancerous and noncancerous origins indicates that the inclusion of an optimal ratio of ATTA in the liposomes substantially enhances the transfection efficiency, superior to that of a traditional liposome, DOTAP–DOPE. Cytotoxicity of ATTA-containing formulations against multiple cell lines indicates potentially distinct activity between cancer and noncancer cell lines. Furthermore, lipoplexes of the ATTA-containing formulations with anticancer therapeutic gene, plasmid encoding tumor necrosis factor-related apoptosis-inducing ligand (pTRAIL), induce obviously more cytotoxicity than conventional formulations. The results indicate that arginine-rich cationic lipopeptide appears to be a promising ingredient in gene delivery vector formulations to enhance transfection efficiency and cell-selective cytotoxicity.

Cimetidine-Based Cationic Amphiphiles for In Vitro Gene Delivery Targetable to Colon Cancer

“…Gene transfection potentials of catalogue relating novel cationic lipids are typically assessed either by without serum or with merely 10% (v/v) serum, as described in several previous studies. , Unfortunately, serum incompatibility is still one of the primary roadblocks to cationic transfection lipid clinical effectiveness . The adhesion to positively charged cationic liposome sites by negatively charged serum proteins is thought to be the cause of cationic transfection lipid’s normal serum incompatibility.…”

Dicationic amphiphiles bearing an amino acid head group with a long-chain hydrophobic tail for in vitro gene delivery applications

“…Natural amino acid-based cationic lipids could carry and transfect the therapeutic nucleic acid to the various cancerous cells. 19,42 The current lipids tested for transfer of the anti-cancer gene, p53, into the glioma and oral cancer cells led to cell death. When these cells were treated with lipid/p53 lipoplexes, cell viability was dramatically less compared to lipid/pCMV-b-gal lipoplexes at a 2 : 1 N/P ratio (Fig.…”

Effect of Methylation of the Hydrophilic Domain of Tocopheryl Ammonium-Based Lipids on their Nucleic Acid Delivery Properties

“…22,23 In recent times, the efficiency of αtocopherol as a delivery system in vivo for the delivery of nucleic acids has been reported. 15,24 It is reported from previous studies that α-tocopherol performed similarly as cholesterol in changing liposomes, i.e., making the liposomes highly protein-induced disruption-resistant, and it as found that this inhibition of protein-induced disruption is more effective with tocopherol compared to cholesterol. Therefore, this made tocopherol-based liposomes more efficient vectors for in vivo gene delivery.…”

“…Moreover, because of their solvent capacity and biocompatibility, vitamin E-based nanomedicines in cancer therapy have been developed. Furthermore, the serum stability and low toxicity of vitamin E, and the presence of physiological pathways of tocopherol transport to various tissues including brain from blood have led to several investigations on systemic delivery applications. , In recent times, the efficiency of α-tocopherol as a delivery system in vivo for the delivery of nucleic acids has been reported. , It is reported from previous studies that α-tocopherol performed similarly as cholesterol in changing liposomes, i.e., making the liposomes highly protein-induced disruption-resistant, and it as found that this inhibition of protein-induced disruption is more effective with tocopherol compared to cholesterol. Therefore, this made tocopherol-based liposomes more efficient vectors for in vivo gene delivery. ,, Together, impressed by DC-Chol applications and the biological importance of tocopherol, we were inspired to design, synthesize, and evaluate four tocopherol-based cationic derivatives, with varying degrees of methylation, AC-Toc (3β-[ N -(aminoethane)­carbamoyl]­tocopherol) hydrochloride), MC-Toc (3β-[ N -( N ′-methylaminoethane)­carbamoyl]­tocopherol hydrochloride), DC-Toc (3β-[ N -( N ′, N ′-dimethylaminoethane)­carbamoyl]­tocopherol hydrochloride), and TC-Toc (3β-[ N -( N ′, N ′, N ′-trimethylaminoethane)­carbamoyl]­tocopherol hydrochloride).…”