Microneedle and Polymeric Films: Delivery of Proteins, Peptides and Nucleic Acids.

In the last 20 years, protein, peptide and nucleic acid-based therapies have become the fastest growing sector in the pharmaceutical industry and play a vital role in disease therapy. However, the intrinsic sensitivity and large molecular sizes of biotherapeutics limit the available routes of administration. Currently, the main administration routes of biomacromolecules, such as parenteral, oral, pulmonary, nasal, rectal and buccal routes, each have their limitations. Several non-invasive strategies have been proposed to overcome these challenges. Researchers were particularly interested in microneedles (MNs) and polymeric films because of their less invasiveness, convenience and greater potential to preserve the bioactivity of biotherapeutics. By facilitating with MNs and polymeric films, biomacromolecules could provide significant benefits to patients suffering from various diseases such as cancer, diabetes, infectious and ocular diseases. However, before these devices can be used on patients, how to upscale MN manufacture in a cost-effective and timely manner, as well as the long-term safety of MN and polymeric film applications necessitates further investigation.

Liposome-loaded polymeric microneedles for enhanced skin deposition of rifampicin.

Methicillin-resistant Staphylococcus aureus (MRSA) is a prevailing bacterial pathogen linked to superficial skin and soft tissue infections (SSTIs). Rifampicin (RIF), a potent antibiotic against systemic and localised staphylococcal infections, faces limitations due to its low solubility. This constraint hampers its therapeutic potential for MRSA-induced SSTIs. To address this, an advanced liposomal system was designed for efficient dermal RIF delivery. Rifampicin-loaded liposomes (LipoRIF) were embedded within polymeric dissolving microneedles (DMNs) to enable targeted intradermal drug delivery. A robust Design of Experiment (DoE) methodology guided the systematic preparation and optimisation of LipoRIF formulations. The optimal LipoRIF formulation integrated within polymeric DMNs. These LipoRIF-DMNs exhibited favourable mechanical properties and effective skin insertion characteristics. Notably, in vitro assays on skin deposition unveiled a transformative result - the DMN platform significantly enhanced LipoRIF deposition within the skin, surpassing LipoRIF dispersion alone. Moreover, LipoRIF-DMNs displayed minimal cytotoxicity toward cells. Encouragingly, rigorous in vitro antimicrobial evaluations demonstrated LipoRIF-DMNs' capacity to inhibit MRSA growth compared to the control group. LipoRIF-DMNs propose a potentially enhanced, minimally invasive approach to effectively manage SSTIs and superficial skin ailments stemming from MRSA infections.

Polymeric in situ forming depots for long-acting drug delivery systems.

Polymeric in situ forming depots have emerged as highly promising drug delivery systems for long-acting applications. Their effectiveness is attributed to essential characteristics such as biocompatibility, biodegradability, and the ability to form a stable gel or solid upon injection. Moreover, they provide added versatility by complementing existing polymeric drug delivery systems like micro- and nanoparticles. The formulation's low viscosity facilitates manufacturing unit operations and enhances delivery efficiency, as it can be easily administered via hypodermic needles. The release mechanism of drugs from these systems can be predetermined using various functional polymers. To enable unique depot design, numerous strategies involving physiological and chemical stimuli have been explored. Important assessment criteria for in situ forming depots include biocompatibility, gel strength and syringeability, texture, biodegradation, release profile, and sterility. This review focuses on the fabrication approaches, key evaluation parameters, and pharmaceutical applications of in situ forming depots, considering perspectives from academia and industry. Additionally, insights about the future prospects of this technology are discussed.

Safe Subunit Green Vaccines Confer Robust Immunity and Protection against Mucosal Brucella Infection in Mice.

Brucellosis is a zoonotic disease that causes significant negative impacts on the animal industry and affects over half a million people worldwide every year. The limited safety and efficacy of current animal brucellosis vaccines, combined with the lack of a licensed human brucellosis vaccine, have led researchers to search for new vaccine strategies to combat the disease. To this end, the present research aimed to evaluate the safety and efficacy of a green vaccine candidate that combines Brucella abortus S19 smooth lipopolysaccharide (sLPS) with Quillaja saponin (QS) or QS-Xyloglucan mix (QS-X) against mucosal brucellosis in BALB/C mice. The results of the study indicate that administering two doses of either sLPS-QS or sLPS-QS-X was safe for the animals, triggered a robust immune response, and enhanced protection following intranasal challenge with S19. Specifically, the vaccine combinations led to the secretion of IgA and IgG1 in the BALF of the immunized mice. We also found a mixed IgG1/IgG2a systemic response indicating evidence of both Th1 and Th2 activation, with a predominance of the IgG1 over the IgG2a. These candidates resulted in significant reductions in the bioburden of lung, liver, and spleen tissue compared to the PBS control group. The sLPS-QS vaccination had conferred the greatest protection, with a 130-fold reduction in Brucella burdens in lung and a 55.74-fold reduction in the spleen compared to PBS controls. Vaccination with sLPS-QS-X resulted in the highest reduction in splenic Brucella loads, with a 364.6-fold decrease in bacterial titer compared to non-vaccinated animals. The study suggests that the tested vaccine candidates are safe and effective in increasing the animals' ability to respond to brucellosis via mucosal challenge. It also supports the use of the S19 challenge strain as a safe and cost-effective method for testing Brucella vaccine candidates under BSL-2 containment conditions.

Hyaluronic acid-coated solid lipid nanoparticles enhance antirheumatic activity and reduce toxicity of methotrexate.

Aim: Methotrexate (MTX) is used to treat rheumatoid arthritis (RA) but is associated with severe toxicity. To minimize these side effects of MTX, the study was undertaken to explore its delivery using solid lipid nanoparticles (SLNs). Materials & methods: MTX-loaded SLNs were synthesized and coated with hyaluronic acid (HA) for targeted drug delivery and evaluated for their safety and efficacy in a complete Freund's adjuvant (CFA) model. Results: HA-MTX-SLNs (230.0 ± 46.4 nm) with 78.75% entrapment were developed and showed sustained drug release with a significant reduction in toxicity and enhanced activity of the drug at the targeted site upon oral administration in CFA-induced rats. Conclusion: HA-MTX-SLNs can be considered as an efficient therapeutic agents for the treatment of RA.

Rheumatoid arthritis (RA) is an autoimmune disease of the joints with no cure and treatment modalities only focus on reducing the symptoms. Methotrexate (MTX) is a primary drug used for its treatment but is associated with severe toxicity. The study aimed to use solid lipid nanoparticles (SLNs) as carriers for MTX to achieve improved efficacy in RA treatment at reduced doses, thus decreasing the potential toxicity of the drug, making SLNs suitable and safe drug carriers. MTX-loaded SLNs (MTX-SLNs) were formulated and coated with hyaluronic acid (HA; HA-MTX-SLNs) and were evaluated for their efficacy in a complete Freund's adjuvant (CFA)-induced arthritic rat model. Both MTX-SLNs and HA-MTX-SLNs demonstrated a significant reduction in toxicity and enhanced the activity of the drug upon oral administration. The HA coating further enriched the antirheumatic activity of MTX, owing to its ability to improve the oral bioavailability and targeted drug delivery of the formulation. Thus, HA-MTX-SLNs can be considered efficient therapeutic agents for the treatment of RA.

Nanoparticle Engineering of Aprepitant Using Nano-by-Design (NbD) Approach.

The aim of this research was to develop a nanosuspension of aprepitant (APT) using the Nano-by-Design approach. A novel microfluidization technology was used for processing the formulation. A 32 full factorial design was used for the optimization of dependent variables, which included critical quality attributes like particle size and polydispersity index. Subsequently, the design space was generated and the optimum formulation was located using desirability constraints followed by its validation.The prepared nanosuspension had a particle size of 721 nm ± 5%, a polydispersity index of 0.106 ± 3%, and a zeta potential of - 8.06 ± 5 mV. Its surface morphology was studied using SEM, DSC, and XRD. It revealed that the prepared nanosuspension had a nano-crystalline nature. The process parameters did not lead to any physicochemical interaction between the drug and excipients. This was confirmed using FTIR analysis. In vitro dissolution studies revealed 100% cumulative drug release over 60 min, showing better results in comparison with pure APT. Thus, it has been shown that microfluidization can be an industrially feasible, novel, green technology for the preparation of a stable APT nanosuspension for improving the dissolution profile of the drug.

Gellan gum-based in situ gelling ophthalmic nanosuspension of Posaconazole.

The formulation and delivery of highly hydrophobic drugs in an optimized dosage form is challenging to formulation scientists. Posaconazole has shown promising action in case studies against fungal keratitis. Biological macromolecules like gellan gum would aid in enhancing the availability of such drugs by increasing the contact time of the formulation. Herein, we propose a transmucosal ocular delivery system of Posaconazole by developing a gellan gum-based in situ gelling nanosuspension. The HPLC method for Posaconazole was developed and validated as per ICH guidelines. The nanosuspension was prepared by microfluidization and optimized by Quality by Design. The gellan gum concentration selected was 0.4% w/v based on the viscosity and mucoadhesion measurements. A greater zone of inhibition of ~ 15 mm was observed for the prepared nanosuspension as compared to ~ 11 mm for the marketed itraconazole nanosuspension. A potential irritancy score of 0.85, considered to be non-irritant, was observed for the developed nanosuspension. Higher drug release of ~ 35% was noted for the nanosuspension compared to about ~ 10% for the coarse suspension. Ex vivo corneal retention studies on excised goat cornea demonstrated ~ 70% drug retention in the tissue. Graphical abstract depicting the central hypothesis of the work.

Nanoparticle-eluting stents for coronary intervention: formulation, characterization, and in vitro evaluation.

Coronary artery disease (CAD) is currently a leading cause of death worldwide. In the history of percutaneous coronary intervention for the treatment of CAD, a drug-eluting stent (DES) is recognized as a revolutionary technology that has the unique ability to significantly reduce restenosis and provide both mechanical and biological solutions simultaneously to the target lesion. The aim of the research work was to design and fabricate DES coated with a nanoparticulate drug formulation. Sirolimus, an inhibitor of the smooth muscle cell (SMC) proliferation and migration, was encapsulated in polymeric nanoparticles (NPs). The NP formulation was characterized for various physicochemical parameters. Cell viability and cell uptake studies were performed using human coronary artery smooth muscle cells (HCASMCs). The developed NP formulation showed enhanced efficacy compared to plain drug solution and exhibited time-dependent uptake into the HCASMCs. The developed NP formulation was coated on the Flexinnium™ ultra-thin cobalt-chromium alloy coronary stent platform. The NP-coated stents were characterized for morphology and residual solvent analysis. In vitro drug release was also evaluated. Ex vivo arterial permeation was carried out to evaluate the NP uptake from the surface of the stents. The characterization studies together corroborated that the developed NP coated stent can be a promising replacement of the current DESs.

Docusate-Based Ionic Liquids of Anthelmintic Benzimidazoles Show Improved Pharmaceutical Processability, Lipid Solubility, and in Vitro Activity against Cryptococcus neoformans.

As the existing therapeutic modalities for the treatment of cryptococcal meningitis (CM) have suboptimal efficacy, repurposing existing drugs for the treatment of CM is of great interest. The FDA-approved anthelmintic benzimidazoles, albendazole, mebendazole, and flubendazole, have demonstrated potent but variable in vitro activity against Cryptococcus neoformans, the predominant fungal species responsible for CM. We performed molecular docking studies to ascertain the interaction of albendazole, mebendazole, and flubendazole with a C. neoformans ß-tubulin structure, which revealed differential binding interactions and explained the different in vitro efficacies reported previously and observed in this investigation. Despite their promising in vitro efficacy, the repurposing of anthelmintic benzimidazoles for oral CM therapy is significantly hampered due to their high crystallinity, poor pharmaceutical processability, low and pH-dependent solubility, and drug precipitation upon entering the intestine, all of which result in low and variable oral bioavailability. Here, we demonstrate that the anthelmintic benzimidazoles can be transformed into partially amorphous low-melting ionic liquids (ILs) with a simple metathesis reaction using amphiphilic sodium docusate as a counterion. In vitro efficacy studies on a laboratory reference and a clinical isolate of C. neoformans showed 2- to 4-fold lower IC90 values for docusate-based ILs compared to the pure anthelmintic benzimidazoles. Furthermore, using a C. neoformans strain with green fluorescent protein (GFP)-tagged ß-tubulin and albendazole and its docusate IL as model candidates, we showed that the benzimidazoles and their ILs reduce the viability of C. neoformans by interfering with its microtubule assembly. Unlike pure anthelmintic benzimidazoles, the docusate-based ILs showed excellent solubility in organic solvents and >30-fold higher solubility in bioavailability-enhancing lipid vehicles. Finally, the docusate ILs were successfully incorporated into SoluPlus, a self-assembling biodegradable polymer, which upon dilution with water formed polymeric micelles with a size of <100 nm. Thus, the development of docusate-based ILs represents an effective approach to improve the physicochemical properties and potency of anthelmintic benzimidazoles to facilitate their repurposing and preclinical development for CM therapy.

Intranasal vaccines for SARS-CoV-2: From challenges to potential in COVID-19 management.

Unlike conventional Coronavirus 2019 (COVID-19) vaccines, intranasal vaccines display a superior advantage because the nasal mucosa is often the initial site of infection. Preclinical and clinical studies concerning intranasal immunization elicit high neutralizing antibody generation and mucosal IgA and T cell responses that avoid severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in both; the upper and lower respiratory tract. A nasal formulation is non-invasive with high appeal to patients. Intranasal vaccines enable self-administration and can be designed to survive at ambient temperatures, thereby simplifying logistical aspects of transport and storage. In this review, we provide an overview of nasal vaccines with a focus on formulation development as well as ongoing preclinical and clinical studies for SARS-CoV-2 intranasal vaccine products.

Solid lipid nanoparticles for hydrophilic drugs.

Hydrophilic drugs are proficient therapeutic agents however, delivery of these drugs is a difficult task. Hence, developing an efficient drug delivery system may require a multipronged approach. Colloidal drug delivery systems such as emulsions, liposomes, nanoemulsions, polymeric nanoparticles, and niosomes are known to enhance drug entrapment, bioavailability, and to improve the pharmacokinetic profiles of hydrophilic drugs. However, issues such as drug leakage and burst release are frequently reported with such systems. Solid lipid nanoparticles (SLNs) were developed as an alternative to the traditional colloidal drug carriers to overcome these issues. Although SLNs have been widely studied as carriers for hydrophobic drugs, delivery of hydrophilic molecules remains a challenge. Hence, the current review focuses on different approaches that have been used for the delivery of hydrophilic drugs using SLNs. It not only discusses various modifications in the traditional methods for the synthesis but also emphasizes modifications of the hydrophilic drugs itself that can help in their efficient entrapment into SLNs drug carriers.

A triple combination 'nano' dry powder inhaler for tuberculosis: in vitro and in vivo pulmonary characterization.

Inhalation route of drug delivery is the most favorable for pulmonary infections wherein direct drug delivery is desired to the lungs. Tuberculosis is one such infection suffering from poor therapeutic efficacy because of low patient compliance due to high drug dosing and lengthy treatment protocols. The current research work was undertaken to develop a dry powder inhaler (DPI) for administration of three first-line antitubercular antibiotics directly to the lungs to improve the treatment rates. Nanoformulations of isoniazid, pyrazinamide, and rifampicin were prepared, spray-dried to obtain a dry powder system, and blended with inhalation grade lactose to develop the DPI. The DPI was evaluated for its flow properties, pulmonary deposition, dissolution profile, and stability. The DPI possessed excellent flow properties with a fine particle fraction of 45% and a mass median aerodynamic diameter of approximately 5 µm indicating satisfactory lung deposition. In vitro drug release exhibited a sustained release of the formulations. In vivo studies showed a prolonged deposition in the lung at elevated concentrations compared to oral therapy. Stability studies proved that the formulation remained stable at accelerated and long-term stability conditions. The DPI could complement the existing oral therapy in enhancing the therapeutic efficacy in patients.

Computational avenues in oral protein and peptide therapeutics.

Proteins and peptides are amongst the most sought-after biomolecules because of their exceptional potential to cater to a vast range of diseases. Although widely studied and researched, the oral delivery of these biomolecules remains a challenge. Alongside formulation strategies, approaches to overcome the inherent barriers for peptide absorption are being designed at the molecular level to establish a sound rationale and to achieve higher bioavailability. Computer-aided drug design (CADD) is a modern in silico approach for developing successful bio-formulations. CADD enables intricate study of the biomolecules in conjunction with their target sites or receptors at the molecular level. Knowledge of the molecular interactions of proteins and peptides makes way for the pre-screening of suitable formulation components and facilitates their delivery.

Gene Therapy for Prostate Cancer: A Review.

BACKGROUND: According to the American Cancer Society, prostate cancer ranks second in terms of mortality and is a front-runner of newly detected cases. Conventional therapies neither eradicated cancer nor increased the life expectancy of patients obviating the need for less toxic as well as efficient therapies to treat cancer. Gene therapy alone, or in combination with conventional therapies, possesses a strong potential to combat cancer. METHODS: This review encompasses a brief note on the etiology and conventional therapy of prostate cancer with an emphasis on gene therapy and its suitability for the treatment of prostate cancer. RESULTS: A comprehensive range of gene therapy approaches have been successfully explored for prostate cancer treatment in animal models and this has been well translated into early clinical trials. We have also discussed in brief about specific therapeutic genes and suitable vector systems for gene therapy in prostate cancer. CONCLUSION: Based on the results of these clinical trials, the application of gene therapy in prostate cancer therapeutics can be satisfactorily established.

Docetaxel Loaded Pomegranate Seed Oil Based Nanostructured Lipid Carriers: A Potential Alternative to Current Formulation.

The current work is focused on the development of docetaxel loaded pomegranate seed oil based lipid nanosystem. Docetaxel loaded pomegranate seed oil nanostructured lipid carriers (DTX-PSO-NLCs) were formulated by the melt emulsification method for parenteral delivery. The developed formulation was characterized in terms of their physicochemical parameters, solid-state characterization, in vitro drug release, in vitro cytotoxicity studies, and in vivo pharmacokinetics and biodistribution studies. Stability studies were carried out as per ICH guidelines Q1A. Melt emulsification method resulted in the formulation of stable DTX-PSO-NLCs with a particle size in the range of 150-180 nm and an entrapment efficiency of 63-65%. The in vitro release showed a slow and sustained release of the drug from the formulation compared to the marketed formulation (i.e., Daxotel®). The formulation was found to be stable for a period of 12 months at conditions of 4°C ± 2°C, 25°C ± 2°C/60% RH ± 5%RH, and 40°C ± 2°C/75% RH ± 5%RH. The developed nanosystem exhibited promising antitumor activity against various types of cancerous cell lines (i.e., MCF7, DU145, U87MG, and NCI-H460) relative to the marketed formulation. The pharmacokinetic evaluation revealed that DTX-PSO-NLCs had a better kinetic profile compared to the marketed formulation. Graphical abstract.

Nose-to-brain delivery: exploring newer domains for glioblastoma multiforme management.

Glioblastoma multiforme (GBM) is the most common and aggressive form of the primary brain tumors in humans. The intricate pathophysiology, the development of resistance by tumor cells, and the inability of the drugs to effectively cross the blood-brain and blood-tumor barriers result in poor prognosis for GBM patients, with a median survival time of only 1 to 2 years. Nose-to-brain delivery offers an attractive, noninvasive strategy to enhance drug penetration or transport novel drug/gene carriers into the brain. Although the exact mechanism of intranasal delivery remains elusive, the olfactory and trigeminal nerve pathways have been found to play a vital role in circumventing the traditional barriers of brain targeting. This review discusses the intranasal pathway as a novel domain for delivering drugs and nanocarriers encapsulating drugs/genes, as well as stem cell carriers specifically to the glioma cells. Considering the fact that most of these studies are still in preclinical stage, translating such intranasal delivery strategies from bench to bedside would be a critical step for better management and prognosis of GBM. Graphical abstract.

Starch microsponges for enhanced retention and efficacy of topical sunscreen.

Topical sunscreen products are universally applied by numerous individuals to protect their skin from the detrimental effects of UV radiation. However, lately, studies have revealed the risks associated with percutaneous absorption of UV filters leading to undesirable systemic side effects such as hormonal disturbances and allergies. In this study, an innovative sunscreen formulation was developed based on starch microsponges as a key carrier encapsulating an organic sunscreen benzophenone­3. The developed starch microsponges were characterized by scanning electron microscopy and nitrogen adsorption/desorption analysis. The results showed that starch microsponges possessed a high BET surface area (85.45 m2/g) with spherical porous morphology with pore size <200 nm. Benzophenone­3 was loaded into the starch microsponges by immersion/solvent evaporation and benzophenone­3 loaded starch microsponges were characterized by scanning electron microscopy, differential scanning calorimetry, powder X-ray diffraction, Fourier transform infrared spectroscopy, and nitrogen adsorption-desorption measurements. Results corroborated that benzophenone­3 was successfully entrapped within the nanopores of starch microsponges. A starch microsponge based sunscreen cream was formulated, characterized and clinically tested. Rheological, texture and sensorial assessment showed that starch microsponges based sunscreen product showed good spreadability, non-sticky, rich texture favorable for consumer usage. In vitro and ex-vivo studies demonstrated benzophenone­3 loaded starch microsponges gave improved photoprotection, higher SPF and reduced cutaneous penetration compared to raw benzophenone­3 cream. Clinically, patch study confirmed that the developed starch microsponges based sunscreen cream was skin safe and biocompatible. Thus, the amalgamation of sunscreen molecule benzophenone­3 into starch microsponges produced a safe, effective innovative sunscreen product.

Preparation and Characterization of Micelles.

Nanoformulations in the past few decades have gained tremendous attention owing to their affirmative applications in increasing the bioavailability of poorly soluble drugs. Micelles in particular are favored due to their varied advantages which include thermodynamic stability, simple formulating steps, Newtonian flow, and enhanced biological barrier penetration. Owing to these advantages micellar nanosystems find extensive applications in oral, transdermal, and parenteral administration, and are now being explored for ocular and other noninvasive novel pathways of drug delivery such as nose to brain. In this chapter, we have discussed the protocol for the preparation of sumatriptan loaded micelles for the therapy of migraine. The inner core of these micelles comprises hydrophobic region of diblock polymer which holds the drug, while the hydrophilic region of the same provides conformational stability in the aqueous environment.

Fabrication of Nanostructured Lipid Carriers (NLC)-Based Gels from Microemulsion Template for Delivery Through Skin.

Nanostructured lipid carriers (NLC) represent the novel and widely explored generation of lipid nanoparticles. These are the second-generation solid lipid nanoparticles (SLN) developed with the aim to overcome limitations of SLN mainly with respect to limited drug loading and drug leakage during its storage. NLC are fabricated by mixing solid lipids with spatially incompatible (liquid) lipids leading to nanoparticulate structures with improved drug loading and controllable release properties. Out of the numerous methods reported to prepare NLC, microemulsion template (ME) technique is the most simple and preferred method. This methodology of preparation of lipid nanoparticles obviates the need for specialized equipment and energy to generate NLC, enables achieving desirable particle size of nanoparticles by modulating the size of the emulsion droplet, and is also feasible for easy scale-up. This chapter describes microemulsion template technique for fabrication of NLC based gel for topical delivery, particularly with respect to its method of preparation and product analysis.

Preparation and Characterization of Solid Lipid Nanoparticles-Based Gel for Topical Delivery.

Solid lipid nanoparticles (SLNs) have been extensively investigated for effective delivery of both hydrophilic and lipophilic drugs by topical route. There are several scalable techniques for the preparation of SLNs such as homogenization, microemulsion template, and solvent emulsification diffusion. This chapter describes step-wise methodology for the preparation and characterization of SLNs using solvent emulsification diffusion method. Tretinoin, a lipophilic entity, was chosen as a model drug. The critical aspects and the important interpretations with respect to the preparation and characterization of SLNs are reported in "Notes" section.