Copper-Free ‘Click’ Chemistry-Based Synthesis and Characterization of Carbonic Anhydrase-IX Anchored Albumin-Paclitaxel Nanoparticles for Targeting Tumor Hypoxia

Abstract

Triple negative breast cancer (TNBC) is a difficult to treat disease due to the absence of the three unique receptors estrogen, progesterone and herceptin-2 (HER-2). To improve the current therapy and overcome the resistance of TNBC, there is unmet need to develop an effective targeted therapy. In this regard, one of the logical and economical approaches is to develop a tumor hypoxia-targeting drug formulation platform for selective delivery of payload to the drug-resistant and invasive cell population of TNBC tumors. Toward this, we developed a Carbonic Anhydrase IX (CA IX) receptor targeting human serum albumin (HSA) carriers to deliver the potent anticancer drug, Paclitaxel (PTX). We used Acetazolamide (ATZ), a small molecule ligand of CA IX to selectively deliver HSA-PTX in TNBC cells. A novel method of synthesis involving copper free ‘click’ chemistry (Dibenzocyclooctyl, DBCO) moiety with an azide-labeled reaction partner, known as Strain-Promoted Alkyne Azide Cycloaddition (SPAAC) along with a desolvation method for PTX loading were used in the present study to arrive at the CA IX selective nano-carriers, HSA-PTX-ATZ. The anticancer effect of HSA-PTX-ATZ is higher compared to HSA, PTX and non-targeted HSA-PTX in MDA-MB-231 and MDA-MB-468 cells. The cell killing effect is associated with induction of early and late phases of apoptosis. Overall, our proof-of-concept study shows a promising avenue for hypoxia-targeted drug delivery that can be adapted to several types of cancers.

Amphiphilic Polymeric Micelles Based on Deoxycholic Acid and Folic Acid Modified Chitosan for the Delivery of Paclitaxel

“…However, the low solubility of PTX due to its bulky polycyclic structure hampers its clinical application [ 1 ]. Many attempts have been made to find less toxic and better-tolerated carriers to increase the solubility of PTX for intravenous delivery, such as nanoparticles, dendrimers, liposomes and nanosuspensions [ 2 , 3 , 4 , 5 ]. In recent years, polymeric micelles have attracted growing interest due to their attractive characteristics, such as their excellent solubilization ability, small size, high stability, prolonged circulation time, low toxicity, ability to evade scavenging by the mononuclear phagocyte system (MPS), high biocompatibility and efficient accumulation in tumor tissues via an enhanced permeability and retention (EPR) effect [ 6 , 7 ].…”

A Biomimetic Drug Delivery System Targeting Tumor Hypoxia in Triple-Negative Breast Cancers

“…Essentially, it is seen to be overexpressed in the core of the solid tumors where there is a shortage of oxygen. Hypoxia is a physiological condition of lack of oxygen that leads to acidosis which induces CA IX on the tumor cell surface that can be easily targeted using a suitable inhibitor like acetazolamide [19,29,33,34]. Moreover, the BSA used for designing this nanoparticle serves as a biomimetic protein that easily circulates in the blood without producing any immunogenic effects.…”

“…The designing of the drug delivery system that is used to target hypoxia in this research paper follows the same procedure as that published in our previous work [29]. The process involves the use of a four-step reaction using a copper-free cyclic Alkyne-Azide click reaction.…”

Development of asialoglycoprotein receptor directed nanoparticles for selective delivery of curcumin derivative to hepatocellular carcinoma

“…HepG2 human HCC cells (5 × 10 4 ) were seeded in six-well plate and incubated at 37 °C under 5% CO 2 for 24 h. The media was replaced with 2 ml of serum-free, antibiotic-free medium and treated with various concentrations of labeled formulations and incubated for 2 h. The formulations containing medium was removed, and resulting cells were washed thrice with PBS and fixed with 2% formaldehyde in the PBS at room temperature for 15 min. Nuclei were stained with a blue fluorescence dye (Hoechst 3342, Thermo Fisher Scientific, Waltham, MA) at RT for 5–10 min and samples were quantitatively analyzed using fluorescence microscopy [52, 53, 54, 55].…”