Biodegradable concanavalin A functionalized polycaprolactone nanoparticle: A promising avenue for cancer therapy.

OBJECTIVE: Receptor-based tumor-selective delivery of therapeutic efficacy and therapeutic index of cytotoxic drugs that exhibit dose-limiting toxicity is observed. Concanavalin A (Con A) was selected as the ligand for the proposed system, which was appended to the polycaprolactone nanoparticles (NPs) carrying the drug to be a very efficient approach for the treatment of cancer. METHODS: Preparation of plain polycaprolactone nanoparticles was carried out employing the emulsion diffusion evaporation technique. Con A was conjugated using carbodiimide chemistry by coupling -COOH group on the surface of nanoparticles. The paclitaxel-loaded Con A-conjugated nanoparticles were further subjected to the characterization of various parameters, that is, surface morphology, particle size, and polydispersity index. In vitro drug release study of both the formulations (plain & conjugated) was done using a dialysis tube up to 48 h in phosphate buffer (pH 7.4). RESULTS: Studies done in xenograft models evidently propose a dose-dependent cytotoxicity response, that is, shrink in % cell growth with increase in the concentration of the drug. The fluorescence photomicrograph clearly revealed the access of the Con A-conjugated nanoparticles to the tumor. A noteworthy biodistribution difference of the paclitaxel from prepared systems was observed. At the same time, Con A-coupled nanoparticles increased the accumulation of paclitaxel in the tumor cells. CONCLUSIONS: Hence, the Con A-conjugated nanoparticles formulation as compared to uncoupled solid lipid nanoparticles formulation and free drug solution showed nearly two times higher uptake because of the lectin receptors on the surface of tumors. Hence, it was envisaged to design polymeric nanoparticles which would be administered intravenously for better therapeutic efficacy.

A Formulation of Paromomycin Sulphate as a Promising Fucose-Conjugated Nanostructured Lipid Carrier for Antileishmanial Drug Delivery.

Environmental and individual risk factors make leishmaniasis an important public health problem. Presently, there are several medicines existing for the cure of leishmaniasis, but a major problem associated with them is their adverse effects. The affinity to the fucose receptor increases the phagocytosis of ligand-bound carriers and simultaneously targets the delivery of the antileishmanial agent. Paromomycin sulphate-bearing nanostructured lipid carriers (NLCs) were formulated by a double emulsion solvent evaporation technique, and then chemistry of ring opening trailed by the reaction of fucose's aldehyde groups was analyzed for conjugation. The NLCs and conjugated-NLCs were examined in terms of average size, entrapment efficiency of drugs, polydispersity index, zeta potential, and in vitro drug release. Hemolytic toxicity was measured on whole human blood, and percent hemolysis by drug-loaded fucosylated-NLCs is reduced from 21.09 ± 1.5% to 5.81 ± 0.9 compared to the plain drug. Macrophage uptake of fluorescein isothiocyanate (FITC)-loaded plain NLCs and fucosylated-NLCs showed that mean FITC measured intensity increases in macrophage cell lines. MTT cytotoxicity assay ensured that NLCs could be beneficial as a biocompatible drug carrier for biomedical and pharmaceutical use. BALB/c mice were used for in vivo studies. Qualitative uptake of fucosylated-NLCs was observed by fluorescence microscopy, and the access of fucosylated-NLCs to the liver was revealed. Similar results were obtained by biodistribution studies. Therefore, fucose-conjugated nanoparticulate carriers can be designed to target macrophages with antileishmanial agents against the Leishmania parasite.

Hyaluronic acid modified pH-sensitive liposomes for targeted intracellular delivery of doxorubicin.

CONTEXT: Surface-modified pH-sensitive liposomal system may be useful for intracellular delivery of chemotherapeutics. OBJECTIVE: Achieving site-specific targeting with over-expressed hyaluronic acid (HA) receptors along with using pH sensitive liposome carrier for intracellular drug delivery was the aim of this study. MATERIALS AND METHODS: Stealth HA-targeted pH-sensitive liposomes (SL-pH-HA) were developed and evaluated to achieve effective intracellular delivery of doxorubicin (DOX) vis-a-vis enhanced antitumor activity. RESULTS: The in vitro release studies demonstrated that the release of DOX from SL-pH-HA was pH-dependent, i.e. faster at mildly acidic pH ∼5, compared to physiological pH ∼7.4. SLpH-HA was evaluated for their cytotoxicity potential on CD44 receptor expressing MCF-7 cells. The half maximal inhibitory concentration (IC50) of SL-pH-HA and SL-HA were about 1.9 and 2.5 µM, respectively, after 48 h of incubation. The quantitative uptake study revealed higher localization of targeted liposomes in the receptor positive cells, which was further confirmed by fluorescent microscopy. The antitumor efficacy of the DOX-loaded HA-targeted pH-sensitive liposomes was also verified in a tumor xenograft mouse model. DISCUSSION: DOX was efficiently delivered to the tumor site by active targeting via HA and CD44 receptor interaction. The major side-effect of conventional DOX formulation, i.e. cardiotoxicity was also estimated by measuring serum enzyme levels of LDH and CPK and found to be minimized with developed formulation. Overall, HA targeted pH-sensitive liposomes were significantly more potent than the non-targeted liposomes in cells expressing high levels of CD44. CONCLUSION: Results strongly implies the promise of such liposomal system as an intracellular drug delivery carrier developed for potential anticancer treatment.

Dual targeted polymeric nanoparticles based on tumor endothelium and tumor cells for enhanced antitumor drug delivery.

Some specific types of tumor cells and tumor endothelial cells represented CD13 proteins and act as receptors for Asn-Gly-Arg (NGR) motifs containing peptide. These CD13 receptors can be specifically recognized and bind through the specific sequence of cyclic NGR (cNGR) peptide and presented more affinity and specificity toward them. The cNGR peptide was conjugated to the poly(ethylene glycol) (PEG) terminal end in the poly(lactic-co-glycolic) acid PLGA-PEG block copolymer. Then, the ligand conjugated nanoparticles (cNGR-DNB-NPs) encapsulating docetaxel (DTX) were synthesized from preformed block copolymer by the emulsion/solvent evaporation method and characterized for different parameters. The various studies such as in vitro cytotoxicity, cell apoptosis, and cell cycle analysis presented the enhanced therapeutic potential of cNGR-DNB-NPs. The higher cellular uptake was also found in cNGR peptide anchored NPs into HUVEC and HT-1080 cells. However, free cNGR could inhibit receptor mediated intracellular uptake of NPs into both types of cells at 37 and 4 °C temperatures, revealing the involvement of receptor-mediated endocytosis. The in vivo biodistribution and antitumor efficacy studies indicated that targeted NPs have a higher therapeutic efficacy through targeting the tumor-specific site. Therefore, the study exhibited that cNGR-functionalized PEG-PLGA-NPs could be a promising approach for therapeutic applications to efficient antitumor drug delivery.

Mannosylated gelatin nanoparticles bearing isoniazid for effective management of tuberculosis.

The mannosylated gelatin nanoparticles (Mn-GNPs) were prepared for the selective delivery of an antitubercular drug, isoniazid (INH), to the alveolar macrophages. The gelatin nanoparticles (GNPs) were prepared by using a two-step desolvation method and efficiently conjugated with mannose. Various parameters such as particle size, polydispersity index, zeta potential, % entrapment efficiency, in vitro drug release, macrophage uptake, in vivo biodistribution, antitubercular activity and hepatotoxicity of plain and Mn-GNPs were determined. The size of nanoparticles (both plain and Mn-GNPs) was found to be in range of 260-380 nm, and maximum drug payload was found to be 40-55%. Average particle size of Mn-GNPs was more, whereas drug entrapment was lesser compared to plain GNPs. The organ distribution studies demonstrated the efficiency of Mn-GNPs for spatial delivery of INH to alveolar tissues. Intravenous administration of INH loaded Mn-GNPs (I-Mn-GNPs) resulted in significant reduction in bacterial counts in the lungs and spleen of tuberculosis-infected (TB-infected) mice and also reduction in the hepatotoxicity of the drug. This study revealed that mannose conjugated GNPs may be explored as potential carrier for safer and efficient management of TB through targeted delivery of INH when compared to plain GNPs and free drug.

Development and characterization of hyaluronic acid-anchored PLGA nanoparticulate carriers of doxorubicin.

A novel hyaluronic acid-poly(ethylene glycol)-poly(lactide-co-glycolide) (HA-PEG-PLGA) copolymer was synthesized and characterized by infrared and nuclear magnetic resonance spectroscopy. The nanoparticles of doxorubicin (DOX)-loaded HA-PEG-PLGA were prepared and compared with monomethoxy(polyethylene glycol) (MPEG)-PLGA nanoparticles. Nanoparticles were prepared using drug-to-polymer ratios of 1:1 to 1:3. Drug-to-polymer ratio of 1:1 is considered the optimum formulation on the basis of low particle size and high entrapment efficiency. The optimized nanoparticles were characterized for morphology, particle size measurements, differential scanning calorimetry, x-ray diffractometer measurement, drug content, hemolytic toxicity, subacute toxicity, and in vitro DOX release. The in vitro DOX release study was performed at pH 7.4 using a dialysis membrane. HA-PEG-PLGA nanoparticles were able to sustain the release for up to 15 days. The tissue distribution studies were performed with DOX-loaded HA-PEG-PLGA and MPEG-PLGA nanoparticles after intravenous (IV) injection in Ehrlich ascites tumor-bearing mice. The tissue distribution studies showed a higher concentration of DOX in the tumor as compared with MPEG-PLGA nanoparticles. The in vivo tumor inhibition study was also performed after IV injection of DOX-loaded HA-PEG-PLGA nanoparticles up to 15 days. DOX-loaded HA-PEG-PLGA nanoparticles were able to deliver a higher amount of DOX as compared with MPEG-PLGA nanoparticles. The DOX-loaded HA-PEG-PLGA nanoparticles reduced tumor volume significantly as compared with MPEG-PLGA nanoparticles.

Stavudine-loaded mannosylated liposomes: in-vitro anti-HIV-I activity, tissue distribution and pharmacokinetics.

Cells of the mononuclear phagocyte system (MPS) are important hosts for human immunodeficiency virus (HIV). Lectin receptors, which act as molecular targets for sugar molecules, are found on the surface of these cells of the MPS. Stavudine-loaded mannosylated liposomal formulations were developed for targeting to HIV-infected cells. The mannose-binding protein concanavalin A was employed as model system for the determination of in-vitro ligand-binding capacity. Antiretroviral activity was determined using MT-2 cell line. Haematological changes, tissue distribution and pharmacokinetic studies of free, liposomal and mannosylated liposomal drug were performed following a bolus intravenous injection in Sprague-Dawley rats. The entrapment efficiency of mannosylated liposomes was found to be 47.2 +/- 1.57%. Protein-carbohydrate interaction has been utilized for the effective delivery of mannosylated formulations. Cellular drug uptake was maximal when mannosylated liposomes were used. MT2 cells treated continuously with uncoated liposomal formulation had p24 levels 8-12 times lower than the level of free drug solution. Further, the mannosylated liposomes have shown p24 levels that were 14-20 and 1.4-2.3 times lower than the level of free drug and uncoated liposomal formulation treatment, respectively. Similar results were observed when infected MT2 cells were treated overnight. Stavudine, either given plain or incorporated in liposomes, led to development of anaemia and leucocytopenia while mannosylated liposomes overcame these drawbacks. These systems maintained a significant level of stavudine in the liver, spleen and lungs up to 12 h and had greater systemic clearance as compared with free drug or the uncoated liposomal formulation. Mannosylated liposomes have shown potential for the site-specific and ligand-directed delivery systems with desired therapeutics and better pharmacological activity.