Cancer

Cancer treatment has been a crucial need of current time and necessitates the need for effective strategies that facilitates superior transfection and endosomal escape ability and enhances intracellular availability of therapeutics. Taking this into consideration, our lab is involved in design and synthesis of novel amphiphilic cationic heterolipids for fabrication of solid lipid nanoparticles, nanostructures lipid complexes, SMEDDs and lipomers for delivery of therapeutics viz. anticancer drugs and gene silencing agents. Our studies have revealed not only a significant enhancement in intracellular uptake but also improved biopharmaceutical issues related to non-site specific drug delivery and reduced treatment associated toxicity. These smart nanocarriers have proven to be promising strategy for better cancer therapy. 

Affibody molecules are a class of engineered affinity proteins with potential for therapeutic, diagnostic and biotechnological applications. These are small 58 amino acid residues with 3 α helices structure and lack disulfide bridges. The original affibody protein scaffold was designed based on the Z domain (the immunoglobulin G binding domain) of protein A. They can bind to a large number of target proteins or peptides with high affinity from micromolar to picomolar range. Because of their high specificity and affinity for its target, rapid biodistribution and tissue penetration as well as rapid clearance of unbound tracer, they seems to be more promising for active targeting in diseases like cancer. Our group is working on developing affibodies against cancer. 

HIV 

Prevention of HIV transmission by sexual route is one of the major challenges all over the globe. Approximately, 34 million people infected with HIV and among them more than 80% were infected by unprotected sexual intercourse (Vaginal/Anal). To overcome this problem, several prophylactic strategies such as sexual abstinence, use of physical barrier (condoms) have been employed. However, all of the strategies mentioned fail to control the prevalence of HIV due to lack of user compliance. Recently, microbicides were introduced as a new prophylactic strategy which provide both physical and chemical barriers against HIV. Several microbicides are in clinical pipeline and among them recently the Tenofovir microbicidal gel has successfully completed the Phase III trials. Our group is working on implementing the traditional system of medicine, by utilizing green technology for extraction of microbicides and ultimately formulating “nanomicrobicide” which not only provide a protective barrier against HIV transmission during intercourse but also improve user compliance due to their low cost and coital independency. 

Malaria

Malaria is one of the most debilitating parasitic infection in several tropical countries claiming one million lives annually. The complete onus of disease control lies on chemotherapy owing to the absence of an effective vaccine. Poor bioavailability and emergence of drug resistant parasite strains reduce the therapeutic potential of current antimalarials. The need of the day is to enhance the pharmacokinetic and pharmacodynamic profiles of current antimalarials through design of appropriate formulations. With this motto, our research group is actively involved in antimalarial drug formulation development. We have developed a myriad of drug delivery systems to enhance the antimalarial potential of various drugs such as artemether, lumefantrine, atovaquone, primaquine, halofantrine, arteether clotrimazole, curcumin, and clindamycin. These include Nanostructured lipid carriers (NLC), nanoemulsions, Self micro-emulsifying drug delivery systems (SMEDDS), suppositories, nanosuspensions, solid dispersions and polymeric hydrogel nanoparticles. Our group has developed NLC which show rapid selective uptake by the malaria infected red blood cells as opposed to the non-infected counterparts. The NLC technology initially developed for parenteral delivery of artemether has been evaluated successfully for intravenous delivery of anti-malarial combinations such as artemether-clindamycin and artemether-lumefantrine achieving a dose reduction of more than 80 % for both the combinations. 

Neurodegenerative disorders

Central nervous system ailments are considered difficult to treat due to ineffective delivery of CNS drugs to specific disease sites. This critical issue arises because of blood-brain-barrier and blood-cerebrospinal fluid-barrier that restricts entry of therapeutic actives across the brain. The consequential low brain bioavailability of drug requires higher doses at frequent intervals which compromise both treatment feasibility and efficacy. In this context, our lab is focusing on development of nanotechnology based treatment modalities for alleviation of neurodegenerative disorders, brain carcinomas and infections which are of prime concern. Particularly, our research is aimed at development of delivery systems based on colloidal nanocarriers, lipid nanoparticles, microemulsions etc. which by virtue of their nanometric size offer enhanced uptake across brain. Further, we are involved in designing targeted nanocarriers exhibiting synergistic uptake across brain barriers owing to their nano size and selective receptor transport. More specifically, we are working in design and synthesis of novel conjugates tagged with ligand molecules as brain transporters and their application towards development of drug loaded smart nanosized delivery systems towards enhancement of drug bioavailability in the brain.  

Radiopharmaceuticals are pharmaceutical compounds that are labeled with radioisotopes and aimed at diagnosis and therapy. Radiopharmaceuticals are composed of a radioisotope and a carrier molecule. After administration radioisotopes emit radiations based on the type of radioisotope and this may further be used for mapping endogenous sites and imaging purposes or in treatment. The research in our group in this area is currently aimed at imaging of different drug molecules post administration in the body by different routes. The method is less tedious than other analytical methods like HPLC, while at the same time having excellent sensitivity. We have already developed and administered technetium labeled drug molecules via intranasal route and received encouraging results. In future we are planning to explore this particular area for diverse dose administration routes and by using other radioisotopes.

Vaccines

Brucellosis is a neglected zoonotic disease that afflicts humans and farm animals globally and is an endemic in many tropical regions. In humans, brucellosis is transmitted via inadvertent consumption of infected dairy products and direct contact with diseased animals, and frequently afflicts veterinarians and animal handlers. Currently available vaccines for animals are based on live attenuated strains that carry the risk of infection and newer more effective vaccines are required, also from the safety point of view. Our lab is working towards the design and development of vaccines for brucellosis towards protection of animals and humans by making use of antigenic cellular components. 

Vaccine adjuvants is another area in which our lab is extensively involved. There are very few approved vaccine adjuvants and therefore, and urgent need to develop safe and effective vaccine adjuvants. The biocompatibility of lipidic nanocarriers, their ability to offer prolonged release of antigen and immunostimulant to the immune system, their nanoscale dimensions render them ideal for vaccine delivery. We are exploring NLCs developed in our lab for vaccine adjuvant activity. We have evaluated the vaccine adjuvant activity of the NLCs administered intraperitoneally by testing with protein antigen. Further work is underway to evaluate the vaccine adjuvant activity along with immunostimulant molecules.

Diagnostics

Diagnostics currently accessible for identification of brucellosis use time consuming culture methods as gold standard and additionally suffer from non-specificity due to cross reactivity, lack of accessibility to remote areas, requirement of skilled personnel and expensive equipment for analysis.  Point of care immunoassays are swift as they can effectively scan various samples in a comparatively less duration of time, are sensitive, specific and offer a great advantage in accurate and fast diagnosis of infectious diseases. Using this technology as a platform, we have fabricated silica based nanosensors capable of specifically detecting anti-brucella antibodies in serological and non-serological samples of afflicted patients. We have successfully designed point of care immunochromatographic testing strips that can detect anti-brucella antibodies in milk samples from farm animals with high sensitivity and specificity.

Tissue Engineering

Wound healing is a highly complex physiological process involving interplay of various cellular and biochemical factors. A few critical stages include inflammation, cell migration, angiogenesis, matrix synthesis, collagen deposition and re-epithelization. Various wound dressings have emerged over the years to assist the process of wound healing that also ensure avoidance of infection. An ideal wound dressing material must keep the wound moist, allow gaseous exchange, inhibit bacterial growth and absorb wound exudates. Also the disappearance of the scaffolds on the wound must mirror the reappearance of healed tissue. Many scaffolds initially fill up the space otherwise occupied by normal tissue and then provide a framework for its regeneration.

We have earlier fabricated scaffolds and films of chitosan-zinc complex. Chitosan apart from being anti-bacterial is biocompatible, biodegradable, hydrating (providing a moist environment to the healing wound). Zinc on the other hand offers protection from microbes and also positively affects the process of wound healing. The in vitro and in vivo studies conducted with these scaffolds have suggested a significant enhancement in healing compared to control. Taking this work forward, we have replaced chitosan with N,O-carboxymethylchitosan (N,O-CMCh), a water soluble derivative of chitosan with better properties (particularly antibacterial) and have successfully reported the antibacterial superiority of N,O-CMCh-Zn complex over Ch-Zn.

We are currently exploring hydrogels as a scaffold system for wound healing. They are appealing mainly because of their closeness to the extracellular matrix of tissues. We have fabricated a composite scaffold with a novel polysaccharide base comprising a drug (antibacterial and wound healer) and keratin. These upon placement on the wound would convert to a hydrogel offering exudates absorption and moist environment.

Microneedles

Microneedles have been reported to be painless and can deliver the therapeutic moieties in a controlled manner with the convenience of self-administration. Our research in this area is focused on fabrication of microneedles for efficient delivery of protein, vaccine and potent therapeutic moieties for systemic and local drug delivery. Exploiting microneedle-based transdermal devices and formulations will have a great impact in future medicine. Our lab is investigating the feasibility of microneedle fabrication, insertion capabilities and mechanical properties of the microneedles.

Coronary Stents

Biodegradable coronary stents as drug delivery systems are reported to be safer and promising device compared to bare metal stents and drugs eluting stents. In our lab, biodegradable stents are being investigated for percutaneous coronary intervention to support mechanical need of healing artery. We are evaluating drug loaded nanoparticles as a promising technology for improved and sustained drug delivery to the diseased vessel walls. Our studies are focused on fabricating biodegradable coronary drug eluting stents combining the benefits of nanotechnology.

Sickle cell anemia 

Sickle cell disease,is a genetic disorder, wherein heme production is largely affected. Hemoglobin S results from a point mutation in HBB, changing the sixth amino acid in the β-hemoglobin chain from glutamic acid to valine (Glu6Val) causing the red blood cells to sickle (form cresent moon shape). Due to this the life span of sickled RBCs is reduced to 20 days. Our group is utilizing traditional Indian ayurvedic literature and including new technologies available to formulate "nanophytomedicine"  which will help improve the life span of sickled RBCs.

Cosmeceuticals

Our group works on topical formulations for various cosmeceutical applications based on herbals like anti-acne, skin repair, anti –aging, follicular delivery,  hair colouring and shampoos. We have optimized green technology based extraction procedures for extraction, purification and isolation of bioactives from herbal sources. We have developed a supercritical fluid assisted technology for extraction of actives from seabuckthorn berries, formulated its nanocarrier systems and evaluated their in vitro and in vivo skin deposition and anti-aging potential. Besides these we have developed numerous nanoformulations using herbal ingredients like mango kernel fat, natural lipids, ellagic acid, genistein, natural polysaccharides  for skin repair, anti aging, skin rejuvenator to name a few. We are currently working on a few industrial projects for developing novel skin and hair cosmetic products for hair colouring and hair growth through follicular delivery.

Supercritical fluid technology

Our lab specializes in supercritical fluid technology for extraction and development of nanosized formulations using polymers and lipids. Solvents mainly used for supercritical fluid extraction are carbon dioxide, water and ethane. Carbon dioxide is used most frequently compared to other solvents, is well established as a green solvent and we have used it extensively for our research and formulation development. The most important parameters that affect the process of extraction are pressure, temperature, solvent used, solute nature and percentage of co-solvent etc. As carbon dioxide is a non-polar solvent it extracts mainly non-polar compounds.

Advantages of employing supercritical fluids are -

1. Beneficial for the thermally labile compounds

2. Fast extraction process

3. Green process

Our present work on supercritical fluid extraction is on extraction of Wrightia tinctoria using supercritical carbon dioxide. This plant show anti-psoriatic activity and reported in many literature. Bio-actives that show anti-psoriatic activity are triterpenoids and flavonoids. We are also working with rapid expansion for supercritical solution (RESS) technology to develop nano sized formulations for malaria therapy and brucellosis prevention.

Planetary ball milling

Planetary ball mills are used wherever the highest degree of fineness is required. Apart from the classical mixing and size reduction processes, the mills also meet all the technical requirements for colloidal grinding and have the energy input necessary for mechanical alloying processes. The extremely high centrifugal forces of planetary ball mills result in very high pulverization energy and therefore short grinding times. The grinding jar is arranged eccentrically on the sun wheel of the planetary ball mill. The direction of movement of the sun wheel is opposite to that of the grinding jars in the ratio 1:1. The grinding balls in the grinding jars are subjected to superimposed rotational movements, the so-called Coriolis forces. The difference in speeds between the balls and grinding jars produces an interaction between frictional and impact forces, which releases high dynamic energies. The interplay between these forces produces the high and very effective degree of size reduction of the planetary ball mill.

Applications-

1. Grinding and mixing of dry powders

2. Mixing of semisolids like creams and ointments

3. Particle size reduction up to nano size (100 nm) without using of organic solvent 

The availability of a large-scale production method is the prerequisite for the introduction of any product to the market. The term Nanomedicine, as coined by NIH, refers to the application of nanotechnology for treatment, diagnosis, monitoring, and control of biological systems. Nanoparticles, by and far the most exploited products of nanotechnology, are defined as submicron particles having size in nanometer dimensions. Numerous nanoparticle-based drug delivery and drug targeting systems have been developed or are currently under development which include different polymeric and metal nanoparticles, liposomes, micelles, quantum dots, dendrimers, microcapsules, cells, cell ghosts, lipoproteins, and many different nanoassemblies. For pharmaceutical purposes, nanoparticles are solid colloidal particles ranging in size from 10 to 1000 nm (1 μm) and consisting of macromolecular materials in which the active principle (drug or biologically active material) is dissolved, entrapped, encapsulated and/or to which the active principle is adsorbed or attached. Often, the production methods for polymeric nanoparticles are only applicable on lab scale. Large-scale methods are not available, or sometimes the equipment cannot be qualified or does not yield a product of a quality that is acceptable to the regulatory authorities. Further, the production method needs to be low cost to make the nanoparticulate product competitive to conventional dosage forms. If the therapeutic benefit or a new nanoparticulate dosage form is limited, the public health systems could tend to go for the cheaper traditional product. A cost-intensive large-scale production method is no problem if the nanoparticulate product is the only way of treatment or if it is distinctly superior in therapy (e.g. more effective in reducing treatment time, hospitalization time of patients, and subsequently reducing the total treatment costs)”. Further, to summarize the current market status of polymeric nanoparticles they have indicated that, “The major obstacles for the introduction or solid polymeric nanoparticles to the market are the status of excipients (e.g., polymers, toxicity, etc.) and lack of large-scale production methods yielding a product of acceptable quality”. Thus, all these literature reports clearly signify that there is an immense requirement to amend the current set of conditions addressing pharmaceutical nanotechnology and either develop new methods with scale-up capability or design new equipments to tackle the scalability issues encountered with the currently employed methodologies. Literature reports describe different ways to prepare nanopaticles at the laboratory scale, but none of the reports indicate the scalability of these methods. Thus there is a dire need to modify these techniques to achieve the goal of producing tailor-made nanoparticles and at the same time handle the restrictions posed at scale up stages. A few reports indicate the scale up of nanoparticles including systems like inorganic nanoparticles, solid lipid nanoparticles,nanoparticles prepared by using supercritical fluid technology, but no method has been reported for the scale up of nanoparticles of pharmaceutical interest.

We have in the past and are currently working on scale-up and process development of nanosuspensions, polymeric nano particles, nano crystals by attempting to break the barriers restricting 'bench-to-bedside' translation, particularly for nano formulations.