Jonathan Pillai

Microfabricated Engineered Particle Systems for Respiratory Drug Delivery and Other Pharmaceutical Applications

Particle Replication in Non-Wetting Templates (PRINT®) is a platform particle drug delivery technology that coopts the precision and nanoscale spatial resolution inherently afforded by lithographic techniques derived from the microelectronics industry to produce precisely engineered particles. We describe the utility of PRINT technology as a strategy for formulation and delivery of small molecule and biologic therapeutics, highlighting previous studies where particle size, shape, and chemistry have been used to enhance systemic particle distribution properties. In addition, we introduce the application of PRINT technology towards respiratory drug delivery, a particular interest due to the pharmaceutical need for increased control over dry powder characteristics to improve drug delivery and therapeutic indices. To this end, we have produced dry powder particles with micro- and nanoscale geometric features and composed of small molecule and protein therapeutics. Aerosols generated from these particles show attractive properties for efficient pulmonary delivery and differential respiratory deposition characteristics based on particle geometry. This work highlights the advantages of adopting proven microfabrication techniques in achieving unprecedented control over particle geometric design for drug delivery.

Novel platforms for vascular carriers with controlled geometry

The first‐generation platforms for vascular drug delivery adopted spherical morphologies. These carriers relied primarily on the size dependence of the enhanced permeability and retention effect to passively target vasculature, resulting in inefficient delivery due to significant variation in endothelial permeability. Enhanced delivery typically requires active targeting via receptor‐mediated endocytosis by surface conjugation of targeting ligands. However, vascular carriers (VCs) still face numerous challenges en route to reaching their targets before delivery. The control of carrier shape offers opportunities to overcome in vivo barriers and enhance vascular drug delivery. Geometric features influence the ability of carrier particles to navigate physiological flow patterns, evade biological clearance mechanisms, sustain circulation, adhere to the vascular surface, and finally transport across or internalize into the endothelium. Although previous formulation strategies limited the fabrication of nonspherical carriers, numerous recent advances in both top‐down and bottom‐up fabrication techniques have enabled shape modulation as a key design element. As part of a series on vascular drug delivery, this review focuses on recent developments in novel vascular platforms with controlled geometry that enhance or modulate delivery functions. Starting with an overview of controlled geometry platforms, we review their shape‐dependent functional characteristics for each stage of their vascular journey in vivo. We sequentially explore carrier geometries that evade reticuloendothelial system uptake, display enhanced circulation persistence and margination dynamics in flow, encourage adhesion to the vascular surface or extravasation through endothelium, and impact extravascular transport and cell internalization. The eventual biodistribution of VCs results from the culmination of their successive navigation of all these barriers and is profoundly influenced by their morphology. To enhance delivery efficacy, carrier designs synergistically combining controlled geometry with standard drug delivery strategies such as targeting moieties, surface decorations, and bulk material properties are discussed. Finally, we speculate on possibilities for innovation, harnessing shape as a design parameter for the next generation of vascular drug delivery platforms.

Comparative study of oral lipid nanoparticle formulations (LNFs) for chemical stabilization of antitubercular drugs: physicochemical and cellular evaluation

Abstract Rifampicin (RIF) and Isoniazid (INH) are two major first-line antitubercular drugs (ATDs) that are typically administered orally, in combination. However, INH-catalysed degradation of RIF under acidic pH environment of the stomach is a major concern related to its oral delivery, and is dramatically accelerated upon further exposure to and interaction with INH. This interaction, in turn, triggers a direct decline in the available RIF dose below the sub-therapeutic level, thereby diminishing its therapeutic efficacy. We hypothesized that encapsulation of both these important ATDs into lipid nanoparticle formulations (LNFs) may help mitigate the acid hydrolysis of RIF, its subsequent interaction with INH and its eventual INH-mediated accelerated chemical degradation in the gastric environment. We further hypothesized that these LNFs would be capable of enhanced uptake and localization into intra-cellular compartments of lung macrophages, thereby potentially targeting the Tb pathogen in its in vivo niche. For this purpose, we evaluated two promising LNFs, viz., solid lipid nanoparticles (SLNs) and nanostructured lipid carriers (NLCs) for encapsulating these ATDs. Here, we report on the design, development and comparative evaluation of SLN and NLC-based lipid formulations of both INH and RIF. Our strategy of nanoencapsulation substantially prolonged encapsulated RIF release and improved its chemical stability in presence of INH in a simulated gastric acidic environment. In vitro cell culture studies showed a well-quantifiable uptake of LNFs in a human alveolar macrophage cell line. Overall, these evaluations provided promising results for establishing the potential of both formulations for TB therapy.

Lipid Nanoparticle Formulations for Enhanced Antituberculosis Therapy

Lipid nanoparticle formulations (LNFs) are on the frontier of the rapidly expanding arena of nanotechnology-based therapeutics. The prospect of improved antitubercular therapy (ATT) using LNFs as drug carriers shows great potential. Numerous hurdles recurrently associated with the use of antitubercular drugs (ATDs) like organ toxicity, variable bioavailability, poor gastrointestinal (GI) absorption, solubility, chemical instability, prevalence of latent infection, and emergence of drug-resistant M. tuberculosis strains may be overcome or mitigated by the use of LNFs for tuberculosis (TB) therapy. The emergence of two newer subtypes of LNFs, namely, solid-lipid nanoparticles (SLNs) and nanostructured lipid carriers (NLCs), further expand the role of these versatile drug carriers in TB treatment. As such, it is anticipated that LNFs will emerge as one of the more promising carrier systems in the near future for the efficient delivery of ATDs to effectively address current issues with their clinical use. This review focuses on the current role of lipid-based nanoparticle formulations in ATT with a special emphasis on two key variants of LNF technology: SLNs and NLCs that are formulated using nanoencapsulation. We also present the rationale and approaches for future translation and successful commercialization of LNFs.

Implantable drug delivery systems: An overview

Abstract Drug delivery systems, capable of offering the flexibility of maintaining pharmacologically effective therapeutic drug levels for extended periods of time while also allowing “dosing-on demand,” would be considered extremely valuable tools in modern medicine. Implantable drug delivery systems (IDDSs) offer physicians the choice of precision delivery, either locally or into systemic circulation, while guaranteeing optimal dosing over the course of treatment. The major advantage of these systems includes targeted local delivery of the drug at a constant and predetermined rate, thereby minimizing dose required and potential side effects, while improving therapeutic efficacy. These systems are particularly useful for diseases requiring long-term therapy or facing challenges with patient compliance, such as cardiovascular disease, tuberculosis, diabetes, cancer, and chronic pain management. This chapter starts with a review of various types of IDDSs, from biomaterial-based to electromechanical systems. Furthermore, design approaches to optimal drug delivery, including methods to tailor drug release profiles and the mechanism of release kinetics are presented. Potential therapeutic applications and biocompatibility-related issues are then briefly discussed. Finally, this chapter concludes with a summary of future perspectives of IDDSs, particularly in their applicability to precision and personalized medicine.

Exploring Packaged Microvesicle Proteome Composition of Chinese Hamster Ovary Secretome

Background: Chinese Hamster Ovary cells (CHO) are the most preferred host cells to meet the increasing demand for high quality ‘human-like’ complex biologics production, but is faced with the challenge of achieving high yield at an affordable price. Secreted proteins critically impact cell growth and product quality and quantity and an integral part of secretome is the packaged microvesicles. In spite of numerous efforts to characterize spent-media proteome, none have identified specific contribution of microvesicles, necessitating further differential analysis of these defined fractions of spent-media proteome, specifically packaged microvesicles. Methods: We have investigated proteome of microvesicles isolated from lag, log, stationary and death phase of CHO batch culture using LC-MS/MS based-proteomic approach to identify proteins that may be involved in regulation of cell growth, viability and productivity in culture. Results: A total of 89 unique proteins were identified in the microvesicles isolated from lag, log, stationary and death-phase of culture; of these only 8.9% were categorized as secretory proteins leaving ~91% proteins of intracellular and non-secretory nature. Microvesicles were observed to contain a number of culture phase-specific proteins which included cell-signaling molecules, transcription and translation regulators and molecular chaperons; many of which are known growth regulators, indicating the potency of microvesicles in regulating culture health. Conclusions: This is the first report of CHO microvesicular proteome and this knowledge is critical in developing rationale design of perfusion process, downstream purification process for rendering improved product stability and also novel cell engineering approaches to maximize growth and improved media formulations to maximize yield and minimize product degradation.

Lipid Nanoparticle Formulations (LNFs): A holistic drug delivery strategy for improving oral bioavailability of Antitubercular Drugs (ATDs)

A QbD and lifecycle management approach to analytical method development and qualification results in a better understanding and fewer failures of analytical methods due to more robust methods which produce consistent, reliable, quality data throughout the lifecycle. This, in turn, leads to less method transfer failures and method “incidents” when used in the routine environment. As the MNC’s are now applying Quality by Design (QbD) to process development, it is now being recognized that this is also the way forward to improve and standardize approach to analytical procedures. The recent focus within the pharmaceutical industry and associated regulatory bodies on QbD approaches to analytical method development are a positive sign that both the industry and regulators are acknowledging the importance of understanding the contribution of measurement uncertainty to drug processes and products. The benefits of applying QbD principles to analytical methods include identifying and minimizing sources of variability that may lead to poor method robustness and ensuring that the method meets its intended performance requirements throughout the product and method lifecycle. The ultimate goal is to highlight that QbD concepts and terminology can be applied to analytical methods and to suggest how adoption of a QbD approach might be used to develop more robust analytical methods and effective control systems. At the same time, these efforts aim to support more advanced regulatory approaches to change management.T leaves of Cajanus cajan has been traditionally used by Indian village people in wound healing, jaundice, hypocholestrolemia and inflammation. The phytoconstituents of C. cajan includes flavonoids and stilbenes. Wound healing is a complex process that includes inflammation, tissue formation, and remodeling. In order to scientifically prove the claimed utilization of the plant, the effects of the extracts were investigated using burn wound model in mice including in vivo antioxidant and antimicrobial activity. Healing was assessed by the rate of wound contraction, period of epithelization and hydroxyproline content. The antimicrobial activity of extract was also studied against bacterial and fungal strain using agar dilution method. In vivo antioxidant activity was performed to understand the mechanism of wound healing potency. The result showed that C. cajan leaf extract has significant wound healing activity as evident from the rate of wound contraction and epithelization. Hydroxyproline content was correlative with the healing pattern observed. C. cajan leaf extract treatment promote up-regulation of pro-inflammatory cytokines TNF-α during early phase of wound healing, inhibit ROS accumulation and exhibit moderate antimicrobial activity. We propose induction of cytokine production as one of the mechanism for acceleration of wound healing by C. cajan leaf extract which may be due to flavonoids and stilbenes.Materials & Methods: This is an open labeled, randomized, prospective study conducted in the Outpatient Department of Orthopaedics, in Dayanand Medical College and Hospital, Ludhiana. A total of 75 patients were enrolled for the study as per the inclusion criteria. Group 1 (n=25) patients were administered Inj. Vitamin D3 (Cholecalciferol) 6 lac IU Intramuscularly (IM) once a month for 3 months. In Group 2 (n=25), Cap Vitamin D3 60000 IU orally was administered once a week for 5 weeks and in Group 3 (n=25), Vitamin D3 granules 60000 IU (per sachet) orally once a week was given for 5 weeks. In addition, all patients were prescribed oral calcium (500 mg twice a day). The intensity of pain was measured using Visual Analogue Scale (VAS) score. The investigations included serum vitamin D3, serum calcium, serum phosphorus, serum alkaline phosphatase, parathyroid hormone, DEXA scan and X-ray of the affected area. The follow up visits were at 3 months and 6 months.A Disease (AD), a common amyloidosis, has no cure yet. Pathological investigations revealed existence of numerous plaques in the Alzheimer’s patients’ brain composed of Amyloid-β (Aβ) peptides, derived from Amyloid Precursor Proteins (APPs). Although the mechanism of plaque formation is unknown, AD is believed to be caused by the aggregation of misfolded Aβ peptides. β-breaker peptides have been developed that disrupt amyloid aggregates (plaques) in vitro as well as in vivo. We have developed two novel and innovative strategies for β-breaker peptide design. First one is based on the concept of β-sheet breaker α/β hybrid peptides (BSBHps); and the second one on the concept of β-Breaker Di-Peptides (BBDPs). We have demonstrated that such well designed peptides are capable of inhibition of amyloid formation of Aβ peptide. Also, they disrupt preformed toxic fibrillar aggregates into non toxic species in vitro. Such peptides may be useful for designing novel drugs against diverse amyloidoses including Alzheimer’s disease, Parkinson’s disease and diabetes type II. Recent advancements in this direction of research will be discussed in the Pharma Summit-2015.M is a leading cause of human deaths affected by parasitic infection in tropical and subtropical reasons. Out of the four plasmodium species responsible for this disease in human Plasmodium falciparum is the most deadly. Due to the widespread resistance of the current antimalarial drugs, intense research efforts are focused on identification of new potent antimalarials. We report here, a structure based drug discovery strategy for design and synthesis of a series of potent and novel triazine based antimalarials. The X-ray structure of Plasmodium falciparum phosphoethanolamine methyltransferase (PfPMT) is used as a target as it is unique to the parasite. Trisubstituted triazine and its anlaogs are produced by an inexpensive three to four step synthesis giving excellent yields. Parasite growth inhibition assays further confirmed the activity of the molecules to be in 5 to 0.8 μM range showing selectivity towards the parasite over mammalian cells. Molecular dynamics simulations on the PfPMT-inhibitor complex shed light on the inhibition mechanism for further optimization of the lead compounds.M is a serious endemic disease in many parts of the world, affecting 5% of the world’s population. About 40% of the world’s population is at stake of malaria. Arteether is a well known antimalarial mainly used in the treatment of cerebral malaria as well as chloroquine resistance malaria. But the main problems associated with arteether are its low solubility (≅17 μg/ml) and ≅40 % degradation in stomach. Due to these limitations, arteether is available only as intramuscular injection. The aim of present study is to formulate the colon targeted matrix tablets for improvement of colonic bioavailability of arteether by enhancing the solubility with cyclodextrin inclusion complex and preventing its degradation in stomach. Inclusion complexes of arteether with β-cyclodextrin in 1:1 molar ratio were prepared by techniques like co-evaporation and spray drying. The prepared complexes were characterized by using H-NMR, FTIR, differential scanning calorimetry, mass spectroscopy and PXRD. Complexation with β-CD in co-evaporation technique showed the maximum enhancement in solubility, i.e., 77.05 folds, in presence of PVP. This complex with better properties was further selected for preparation of colon targeted matrix tablets to prevent the degradation of arteether in stomach with improved solubility and better colonic bioavailability.A various classes of drugs bisphosphonates (alendronate and risedronate) offers the most potent types of molecules to treat osteoporosis. The major obstacle that limits the successful use of orally administered risedronate includes low permeability and more importantly, insufficient and fluctuating bioavailability. This poor bioavailability (<1%) of risedronate results in the supplementation of high doses that may perhaps lead to severe side effects like osteonecrosis of the jaws, fever, vein irritation, general aches and pains and kidney dysfunction. Therefore, in the present study, bioceramic (hydroxyapatite) based Poly(D,L-Lactide-CoGlycolide) (PLGA) and Polyethylene Glycol (PEG) nanoparticles of risedronate, was prepared by dialysis method for bioavailability enhancement. The structure of prepared diblock copolymers were characterized by FT-IR and NMR spectrometry. The formation of surface-modified PLGA nanoparticle prepared with various ratios of risedronate as well as hydroxyapatite and mPEG was confirmed by 1H NMR and FT-IR spectrometry. Pharmacokinetic study was also performed in male Wistar rats in order to evaluate the efficiency of prepared nanoparticles on existing marketed preparation (rosofos tablet). The size and % entrapment of the prepared nanoparticle was found to be 79.3±2.3 nm and 93±3.1%. Transmission Electron Microscopy (TEM) revealed that mPEG-PLGA-RISHA nanoparticles possess smooth and uniform surface. Pharmacokinetic study showed a significant enhancement in bioavailability (2 fold) when compared to marketed preparation. The results strongly implicated that mPEG-PLGA-RIS-HA has a therapeutic benefits over risedronate sodium monotherapy for the treatment of osteoporosis in a rat model. This research is likely to be helpful in the design of functional nanoparticles for the site-specific drug delivery in the treatment of bone diseases.A drug reaction has been defined as ‘any noxious change which is suspected to be due to a drug, occurs at doses normally used in man, requires treatment or decrease in dose or indicates caution in the future use of the same drug. Our aim of study was to evaluate the incidence of adverse drug reactions in a tertiary care hospital in tertiary care hospital in India. Study was carried out for a period of one year. Patients admitted in wards were taken into study. Total ADR obtained were 130. Males (81) were affected more than females. Most common system involved was haemopoetic system (32%) following by GIT (30%). Most common agents causing were antineoplastic drugs (42%) following by antimicrobials. Type A (118) reactions were most common and most of the ADR fell in possible category. Most of the reactions were mild (53) in category followed by severe (46).