From the President From the Editor Technical Insights Nanoparticle-cell interactions Sameer Jadhav …………………………………....................1 Supercritical Fluids: Simplifying Nanotechnology Megha Swami and Vandana Patravale ……………………..5 Drug-Nutraceutical Interactions: A Revelation Ankita Pai, Lalit Kagliwal, Rekha Singhal……………………….….9 Measuring Yardstick FTIR Imaging In Bio- Medical Technology and Pharmaceutical Industry Purnima Parkhi …………………………………..................13 Patent law and litigation: A comparative perspective of Indian and American doctrines Madhulika Vishwanathan, Yagna Praveen Kumar and Mahalaxmi Andheria ………….………………………………………20 Business Snapshot Commercialization – a Catalyst for Drug Discovery and Innovation Jitendra N Verma ...................................................24 Colloquially Speaking PharmaSearch PharmaScramble PharmaRebus PharmaDouble PharmaCrossword PharmaCrypt CRS News

Volume 5. February 2012

Vandana B. Patravale Chief Editor

Clara Fernandes Associate Editor

Imran Ahmad Khan Associate Editor

Megha Swami Associate Editor

Anuradha Pol Associate Editor

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Glimpses of 11th International Symposium on “Advances in Technology and Business

Potential of New Drug Delivery Systems” 17th and 18th Feb. 2011

at Hotel ITC Grand Maratha, Mumbai.

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Distinguished panel members inaugurating abstract book at 11th International Symposium on Advances in

Technology and business potential of new drug delivery systems

Distinguished panel members inaugurating CRS Newsletter at 11th International Symposium on

Advances in Technology and business potential of new drug delivery systems

Prof. Maitra at the Lamp Lightning Ceremony at 11th International Symposium International Symposium on Advances in Technology and business potential of new drug delivery systems

Prof. Maitra delivering Key Note Address at 11th International Symposium on Advances in Technology and business potential of new drug delivery systems

Section of Audience attending a session at 11th International Symposium on Advances in Technology and business potential of new drug delivery systems

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Welcome to yet another exciting Controlled Release Society - Indian Chapter 2012!!! The pharmaceutical industry is ushering into the defining chapter of its existence. In recent times, the fierce competitiveness from the allied fields has compelled the industry to diversify its research interests. Further, the uncertainty grappling the pharmaceutical market has made INNOVATION pivotal for the longevity of the industry. Although, the academia has embraced this fact, the industry at large is still lagging behind in acclimatizing to this reality. Nevertheless, this dismal scenario is proving to be a catalyst in overhauling the entire outlook of the pharmaceutical community. Innovation, however, should not be restricted to but has to transcend the ambit of drug delivery technologies. It should be borne in mind that there are myriad of opportunities waiting to be explored; especially in newer avenues such as biopolymer synthesis, gene drug delivery, medical device, vaccine, diagnostics and theranostics. Further, the revolution of nanotechnology has made it essential to explore newer technologies for ease of fabrication of these versatile components at industrial level. These favorable prospects have necessitated the need for upgradation and lateral thinking of inventive ways to bolster every quarter of the industry i.e., from research to commercialization. Since its inception, Controlled Release Society - Indian Chapter has zealously pursued the goal of dissemination of the current and innovative knowledge for edifying the intellectual minds of the scientific community. This year too is not an exception, the Controlled Release Society - Indian Chapter through the conference and the Newsletter has made conscientious efforts to introduce the delegates to the recent advancements in the scientific arena. Likewise, the editorial committee has also invested commendable efforts in collating information on the most relevant topics in sync with the conference ideology. Through this Newsletter, the organization intends to encourage the young minds and my fellow colleagues to fearlessly tackle the challenges of the emerging future which holds the promise of only rich dividends. As always, we earnestly hope that the efforts of the editorial team provide impetus to our colleague to think innovative! Finally, I would like to conclude with the motivating quote of former President Abdul J. Kalam ‘Look at the sky. We are not alone. The whole universe is friendly to us and conspires only to give the best to those who dream and work.’ Dr. AMARJIT SINGH PRESIDENT – CRS INDIAN CHAPTER

D Dr. Tejas Gunjikar Showcasing their technology at 11th International Symposium International Symposium on Advances in Technology and business potential of new

drug delivery systems

Poster Presentation at 11th International Symposium on Advances in Technology and business potential of new

drug delivery systems

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Prof. Vinod Labhasetwar addressing the delegates at International Symposium on Advances in

Technology and business potential of new drug delivery systems

Dr. Manish Rane, Colorcon Asia Pvt. Ltd. Showcasing their technology at 11th International Symposium International Symposium on Advances in Technology and business potential of new drug delivery systems

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The Twelfth International Symposium on 'Advances in Technology and Business Potential of New Drug Delivery Systems' holds special relevance to the CRS Newsletter editorial team. Recalling the last few years, a familiar sense of déjà vu grips us as CRS Newsletter celebrates its 5th glorious year of publication. Although still in its infancy, the endeavor has been gradually gaining recognition among the patrons of the CRS Indian Chapter for its attempt to propagate the ideology of innovativeness in outlook towards research. In the past few decades, the pharmaceutical research has transcended from just being mediocre to something more avant-garde. However, this phenomenal change in the knowledge sector has only contributed to further the divide in the academia and industrial doctrines. Ironically, it is the same knowledge which holds the key to dissipate this issue. As daunting as it will be, embracing this change is the only deciding factor for Indian pharmaceuticals to metamorphose into world class leading brands. This changing paradigm is the underlying theme of the 5th edition of CRS Newsletter. Addressing this change is the eclectic mix of distinguished authors who have contributed insightful viewpoints on an array of topics. For their unflagging and timely support to this endeavor, the editorial team expresses heartfelt gratitude to all our immensely busy and talented authors. In recent times, relatively ambiguous nanotechnology has been panned as well as extolled for its virtues. However, the ongoing research has made us realize that there is more to this technology than meets the eye. Explaining what make nanomedicine pivotal in disease management is the feature on Nanoparticle and cell interactions which succinctly elucidates the possible interactions of nanoparticles at the cellular level. While debating on the latest trend of exploiting nature’s bounty is a brief insight on the possible impact of natural antioxidants on the ADME of concomitant administered drugs under the ambit of Drug-Nutraceutical Interactions. Illustrating change is the introductory article on the widely acclaimed green technology; Supercritical fluid: simplifying nanotechnology. Although in its nascence, this technology is being touted as one of the possible alternatives to industrialize nanotechnology. Keeping pace with the innovations is the feature on patents which promises to be an insight on the significance of recognition of the opportunities and risks in patenting innovations. Bringing us to reality is the recount of the one of the most successful entrepreneur in the Indian scenario. This article is an attempt of abreast the young and aspiring entrepreneurs about the possible struggles arising due to the lacunae in the system. Adding to the much needed twist to ubiquitous Infra Red analytical technique is an interesting feature on the imaging analytical technique for facilitating better understanding of the drug and excipients interactions in drug delivery systems. Finally, as always we have lined up brain teasers for providing a breather from these topics. Reiterating the fact, the realization of this issue of Newsletter is an enriching and exhilarating experience for the entire editorial team. We hope we are successful in challenging our readers to think innovative. We strive to excel in our endeavor and we believe attaining excellence is impossible without your feedbacks. So looking forward to your encouraging feedbacks and hoping all of you have a great networking experience. Vandana B. Patravale Professor of Pharmaceutics, ICT, Mumbai.

Newsletter articles reflect only the views of the authors. Publication of articles with in the CRS Newsletter does not constitute endorsement by CRS or its agents of products, services or views expressed herein. No representation is made as to accuracy hereof and the publication is printed subjected to errors and omissions All the E-mail contributions, views and suggestions, should be directed to the newsletter editor at vb.patravale@ictmumbai.edu.in

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1

Introduction

It is close to half a century since the use of nanoscale phospholipid vesicles or liposomes was proposed for drug delivery purposes. Since then nanoparticles made of various organic and inorganic materials have been successfully used for diagnostic and drug delivery applications. Recent nanoparticulate drug delivery systems have been designed for increased therapeutic efficacy. This is achieved via targeted delivery of the drug to the afflicted site and its controlled release, thereby reducing absolute concentration as well as fluctuations in the concentration of the drug. Nanoparticle based systems have proven to be very effective for delivery of adenoviruses, proteins and siRNA. The use of nanoparticles has also been demonstrated for applications such as in vitro diagnostics and in vivo imaging. Moreover, enhancement of therapeutic techniques such as X-ray treatment by gold-nanoparticles and hyperthermia with magnetic nanoparticles has also been achieved [1]. Furthermore, theranostic applications have been proposed where diagnostic and therapeutic actions are simultaneously employed [2]. For example, liposomes enclosing quantum-dots and the cytotoxic drug doxorubicin, have been used for imaging and treatment of tumors [3]. Apart from drug delivery and diagnostics, nanoparticles have also been used in the design of novel biomaterials for tissue engineering applications [1].

Types of nanoparticles

Based on the materials used nanoparticles may be classified as: (a) lipid-based nanoparticles such as liposomes, micelles, (b) polymer-based nanoparticulate dendrimers, hydrogels, polymerosomes, made of polylactic acid, poly-caprolactone, poly-(ethylene glycol), polylactic-coglycolic acid etc. (c) metal-based nanoparticles made of gold, silver etc. (d) metal-oxide nanoparticles made of iron oxide, silica, titania etc. [1, 4]. In addition to these classes there are several hybrid nanoparticulate systems such as liposomes and hydrogels that encapsulate quantum-dots or adenoviruses as well as core-shell nanoparticles with a metal core and a polymer shell [2, 3, 5]. Targeting of the nanoparticles to specific tissue/cells is achieved by conjugation with ligands such as antibodies and adhesive peptides which bind specifically to cell surface receptors [4]. New reports of novel nano-materials with more varied biomedical applications continue to appear in scientific literature with increased frequency.

Barriers to nanoparticle uptake

The route of drug administration is critical for the design of nanocarriers. There is a need for strategies to overcome physical barriers to transport of nanoparticles such as the skin in transdermal route, endothelium in the case of vascular delivery and epithelium for gastro-intestinal and respiratory administration [6]. The mucus lining the enteric route is the first obstacle to nanoparticle transport, and maybe circumvented by alternating arrays of positive and negative charges on the nanoparticles [7]. Rapid clearance of nanoparticles from the blood and lymphatic vessels by the reticulo-endothelial system has been inhibited by poly-electrolyte brushes grafted on the nanoparticle surface. In the case of drugs administered to the central nervous system, a possible strategy to breach the blood-brain barrier may be via caveolae-mediated transcytosis [8]. To enhance nanoparticle transport through solid tissues such as tumors, it has been suggested that the permeability of the extracellular matrix may be increased [9].

Mechanism of nanoparticle uptake

Based on the nature of target cells and their response to the nanocarriers, cellular uptake of nanoparticles is primarily accomplished by either phagocytosis or pinocytosis (Fig. 1). During phagocytosis various opsonins such as immunoglobulins and complement components from the blood plasma adsorb on the nanoparticle surface which are then recognized by receptors on phagocytic leukocytes [10]. This receptor-binding and clustering leads to signal transduction mediated by small GTPases and their downstream effectors which regulate the local reorganization of the actin cytoskeleton beneath bound nanoparticles. The resulting membrane protrusions engulf the nanoparticles into phagosomes which are actively transported into the cell [10]. In contrast to phagocytosis which is limited to certain cell types, pinocytosis is carried out by almost all cell types. The pinocytic pathways primarily consist of clathrin-mediated endocytosis, caveolae-mediated endocytosis and macropinocytosis [11]. Receptor-dependent clathrin-mediated endocytosis is initiated by receptor-ligand clathrin-rich membrane regions form clathrin-coated pits which then close at the mouth to form vesicles. These vesicles are subsequently transported into the cell and end up in lysosomes. A similar vesicle formation by clathrin cages is also observed in the case of receptor-independent clathrin-mediated endocytosis.

Sameer Jadhav, PhD (Johns Hopkins U.) Assistant Professor,

Department of Chemical Engineering, Indian Institute of Technology Bombay, Powai, Mumbai 400076.

Email: srjadhav@che.iitb.ac.in

Nanoparticle-cell interactions

Technical Insights

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Figure 1. Nanoparticle uptake by biological cells. (A) Opsonization and phagocytosis of nanoparticles, (B) receptor- dependent or independent clathrin-mediated pinocytosis, (C) caveolae-mediated pinocytosis and, (D) macropinocytosis.

Caveolae-mediated endocytosis is observed particularly in endothelial cells, smooth muscle cells and fibroblasts. Caveolae are membrane invaginations, lined by the dimeric protein, caveolin. Caveolae-mediated endocytosis results in transport to caveosome thereby avoiding the acid degradation typically observed in lysosomes [10]. Macropinocytosis is another pinocytic process whereby the large membrane protrusions envelope contents at the cell surface leading to formation of macropinosomes which are speculated to subsequently fuse with lysosomes [11].

Nanoparticle interactions with host cells

To assess the biocompatibility of nanoparticles it is typically recommended that they do not exhibit hemolytic activity, thrombogenicity and complement activation [12]. Hemolytic activity is a measure of the ability of a foreign body to induce rupture of the erythrocyte plasma membrane. Thrombogenicity is the property of the foreign material to initiate platelet aggregation leading to blood coagulation. Complement activation is the mechanism by which the immune system of the host responds to "non-self" materials via leukocyte action. The nanoparticles may directly interact with the respective cells or through adsorbed plasma proteins that may result in the activation of thrombogenic or inflammatory processes. Nanoparticle size and shape have been shown to influence the kinetics of their uptake by various cells, with sizes between 25-75nm and spherical nanoparticles exhibiting faster uptake rates [11]. Surface charge and hydrophobicity of nanoparticles has been known to support their adhesive interactions with plasma proteins, cells surface receptors as well as the cell plasma membrane [12, 13]. Grafting neutral and hydrophilic polymers such as poly (ethylene glycol) on nanoparticles has been shown to significantly reduce their non-specific binding to cells and proteins. Polymer-grafted nanoparticle interactions with cells or plasma proteins are very sensitive to polymer charge, size and configuration. For instance, in contrast to dendrimers of neutral polymers, those comprising of cationic polymers were found to disrupt the plasma membrane [13]. Studies also demonstrated that disruption of lipid bilayers by cationic particles occurs

regardless of shape, chemical composition, deformability, charge density, or size [11]. It has been reported that nanoparticles cause local surface reconstruction of lipid bilayers. Specifically, a local gelation was observed on binding of anionic nanoparticles to a fluid lipid bilayer, while positively charged nanoparticles induced fluidity in gel-phase bilayers [14].

Forces at the nano-bio interphase

In order to gain a better understanding of nanoparticle adhesive interactions with cells, it is important to obtain quantitative estimates of the Van der Waals, solvation, depletion and electrostatic forces that act between nanoparticles and the cells (Fig. 2). The adhesive forces between nanoparticles and specific cell types have been shown to be modulated by the presence of certain plasma proteins [15]. Therefore it is also necessary to quantify the forces that act between nanoparticles and absorbed proteins as well as those acting between adsorbed plasma proteins and cell surface receptors. These interactions at single macromolecule level have been measured using atomic force microscopy(AFM) [16]. While scanning probe microscopy enabled imaging the orientation of nanoparticles on cell surfaces, time lapse AFM has been used to estimate the rate of nanoparticle internalization from the cell surface. Using these techniques, it has been reported that functionalized nanoparticle uptake at the surface is significantly faster than that of non-functionalized nanoparticles [17]. Simultaneous imaging and force measurements have also been carried out to understand the spatial variation of receptor density and nanoparticle adhesive interactions at the cell surface [18]. Using laser scanning confocal microscopy and AFM it was shown that nanoparticles with random distribution of hydrophobic patches ended up in lysosomes. In contrast sub-nanometer patterned arrangement of the hydrophobic patches led to their penetration of the plasma membrane without bilayer disruption [19]. Figure 2. Forces relevant to nanoparticle-cell interactions. (A) Van der Waals (electrodynamic) attraction, (B) electrostatic attraction or repulsion, (C) steric repulsion between polymer brush on nanoparticle and cell surface glycocalyx, (D) hydrophobic attraction between nanoparticle and protein (opsonin) and, (E) depletion attraction due to exclusion of large molecules near the cell surface.

Technical Insights

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In the case of active targeting, the avidity of a specific receptor-ligand pair, the affinity between the nanoparticle and target cell as well as the kinetics of receptor-ligand binding/unbinding regulate efficiency of drug delivery. For instance, enhanced in vivo efficacy of nanoparticles over free drug was observed due to multivalent interaction between the multiple ligands on the dendrimeric nanocarrier with multiple high-affinity receptors on the tumor cell surface [20]. Importantly, surface plasmon resonance showed that the on-rate was found to increase linearly with the number of targeting agents and showed no cooperativity, whereas the off-rate, decreased exponentially with the number of targeting agents. Therefore, improved efficacy of treatment was attributed to substantial enhancement of dissociation constant for receptor-ligand interactions and

not an increased rate of endocytosis [20].

Conclusions and future directions

Taken together, these recent developments in nanoparticle synthesis and new probing techniques have provided new insights on the possible mechanisms of nanoparticle recognition and uptake by biological cells. However, the understanding of the processes at the nano-bio-interphase is far from complete. Much work still remains to be done for engineering smart nano-carrier systems that overcome the obstacles related to immuno-compatibility and cell/organ specific targeting. Future progress will depend on novel experimental techniques to probe cell-nanoparticle interactions at a subnanoparticle /sub-macromolecule level.

References: 1. Shi, J.J., et al., Nanotechnology in Drug Delivery and

Tissue Engineering: From Discovery to Applications. Nano Letters, 2010. 10(9): p. 3223-3230.

2. Caldorera-Moore, M.E., W.B. Liechty, and N.A. Peppas, Responsive Theranostic Systems: Integration of Diagnostic Imaging Agents and Responsive Controlled Release Drug Delivery Carriers. Accounts of Chemical Research, 2011. 44(10): p. 1061-1070.

3. Al-Jamal, W.T. and Kostarelos K. , Liposomes: From a Clinically Established Drug Delivery System to a Nanoparticle Platform for Theranostic Nanomedicine. Accounts of Chemical Research, 2011. 44(10): p. 1094-1104.

4. Koo, O.M., Rubinstein I. , and Onyuksel H. Role of nanotechnology in targeted drug delivery and imaging: a concise review. Nanomedicine, 2005. 1(3): p. 193-212.

5. Singh, R. and Lillard J.W. Jr., Nanoparticle-based targeted drug delivery. Exp Mol Pathol, 2009. 86(3): p. 215-23.

6. Kateb, B., et al., Nanoplatforms for constructing new approaches to cancer treatment, imaging, and drug delivery: what should be the policy? Neuroimage, 2011. 54 Suppl 1: p. S106-24.

7. Cone, R.A., Barrier properties of mucus. Advanced Drug Delivery Reviews, 2009. 61(2): p. 75-85.

8. Fernandes, C., U. Soni, and V. Patravale, Nano-interventions for neurodegenerative disorders. Pharmacol Res, 2010. 62(2): p. 166-78.

9. Groothuis, D.R., The blood-brain and blood-tumor barriers: a review of strategies for increasing drug delivery. Neuro Oncol, 2000. 2(1): p. 45-59.

10. Hillaireau, H. and Couvreur P. Nanocarriers' entry into the cell: relevance to drug delivery. Cell Mol Life Sci, 2009. 66(17): p. 2873-96.

11. Verma, A. and F. Stellacci, Effect of surface properties on nanoparticle-cell interactions. Small, 2010. 6(1): p. 12-21. 12. Dobrovolskaia, M.A., et al., Preclinical studies to

understand nanoparticle interaction with the immune system and its potential effects on nanoparticle biodistribution. Mol Pharm, 2008. 5(4): p. 487-95.

13. Leroueil, P.R., et al., Nanoparticle interaction with biological membranes: does nanotechnology present a Janus face? Acc Chem Res, 2007. 40(5): p. 335-42.

14. Wang, B., et al., Nanoparticle-induced surface reconstruction of phospholipid membranes. Proc Natl Acad Sci U S A, 2008. 105(47): p. 18171-5.

15. Karmali, P.P. and D. Simberg, Interactions of nanoparticles with plasma proteins: implication on clearance and toxicity of drug delivery systems. Expert Opin Drug Deliv, 2011. 8(3): p. 343-57.

16. Nel, A.E., et al., Understanding biophysicochemical interactions at the nano-bio interface. Nat Mater, 2009. 8(7): p. 543-57.

17. Vasir, J.K. and Labhasetwar V. , Quantification of the force of nanoparticle-cell membrane interactions and its influence on intracellular trafficking of nanoparticles. Biomaterials, 2008. 29(31): p. 4244-52.

18. Kada, G., Kienberger F. and Hinterdorfer P. Atomic force microscopy in bionanotechnology. Nano Today, 2008. 3(1-2): p. 12-19.

19. Verma, A., et al., Surface-structure-regulated cell-membrane penetration by monolayer-protected nanoparticles. Nat Mater, 2008. 7(7): p. 588-95.

20. Hong, S., et al., The binding avidity of a nanoparticle-based multivalent targeted drug delivery platform. Chem Biol, 2007. 14(1): p. 107-15.

Technical Insights

U B Y P N B I M P X E L P I S B E J T S H D K S R K R X C U T X X S E B I L V B I B G X T O A G F U K I E L W S M Q C X S E E V X J L E H Y H M L P X W F T Y X N J C G X L P I Q S O B P P K L N P W O A N R Q L L Z S N S E L O R P P N L W H N T O U Z E L V A D S R R R N N N P T S O Y F V E C T V X G A L D A A W P M F E Q R G G T O O P S H L C Y T N M A X S L U P A S R N J N L Q O C H S O C U A W C A Y M S A A K X Q W R E Q O S F C O W I D J M R N N M V E W U V F I O O I S K G N N C E S A B R A X A N E R M Z L B S A G H O M D D L A P N R T C P E G A C J C I S V I R S E L C I T R A M S S M D X L J N K D U H U Q U K X C X Y T X E Z Z Q L M B I G N A N O E D G E M R B L G B D X P R V O Y P X S K Y S V V R T L H Q U S H R A S U D E M X Z D V S I N A O Q L

A N D V W J B G S G Q J B U A Z R W E Z

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ABRAXANE, AUROLASE, AVIDIMERS, CAGICLES, HYDROPLEX,IVECT, MAGFORCE, MEDUSA, NANOCELL, NANOEDGE, NANOQUAD, NANOSOMES, NOVASOMES, PAUCILAMELLAR, PRIOSTAR, PROLINDAC,

SIPLEX, SMARTICLES, TRANSDRUG

PharmaWord Search

Nowadays, a great interest in expertise employing renewable resources including green technology has gained great attention due to both technological and environmental issues. In this sense, the pursuit for supercritical fluid technology for nanoengineering, extraction of actives and fabrication of tissue scaffolds has been largely intensified, as the main requirement of medical and pharmaceutical industries.

The properties that make supercritical fluids particularly attractive, as a rule, are gas-like diffusivities, low viscosity, the continuously tunable solvent power/selectivity and the possibility of complete elimination at the end of the process. Supercritical fluid technology will allows pharmaceutical and nutraceutical companies to develop products of standardized concentration of active ingredients, and will simultaneously produce nutraceutical and pharmaceutical products of much higher concentration (higher yields and purity) and quality (with less creation of artifacts), than possible by conventional chemical engineering unit operations. In 1879, Hannay and Hogarth discovered the ability of supercritical fluids to dissolve low-pressure solids. Research has led to several important applications of supercritical fluids including pharmaceutical productions, biological waste disposal and semiconductor. The unique properties of supercritical fluids are due to the existence of a single phase beyond the critical point. Usually, Supercritical carbon dioxide (sc-CO2) is used as a main solvent as it has a low critical temperature (31.1oC) and a moderate critical pressure (73 bar). Moreover, carbon dioxide is naturally abundant, inexpensive, leaves no toxic residue, inert and is non- flammable. In addition to being a solvent for extraction and fractionation (purification) of organic compounds, carbon dioxide is increasingly being utilized as a medium for reactions, as a micronizing agent in Rapid Expansion in a Supercritical Solution process (RESS), as an anti-solvent for crystallization in Gas Anti-Solvent process (GAS), and as a carrier solvent for coating and depositing materials onto or into a solid matrix. SCF is one of the fastest growing new process technologies being adopted by the food, pharmaceutical and nutraceutical industries.

Particle engineering and drug formulation

Particle design process using supercritical fluids are now the subject of increasing interest, especially in the pharmaceutical industry. Here the technology is being applied in various ways, including, to increase

bioavailability of poorly soluble drug, to design formulation for sustained release and to develop less invasive alternatives to parenteral drug delivery (oral, pulmonary and transdermal). The most complex challenge relates to the therapeutic proteins, as it is extremely difficult to deliver bio-molecules due to their instability, and short half life in vivo. In RESS process, a supercritical saturated solution of a dissolved solute is rapidly (~10-6 sec) depressurized across a heated micro-orifice (nozzle). With the dramatic decrease in solvent density, fine particles are formed under highly supersaturated conditions and then quickly quenched. It is an attractive technology for the production of small, uniform and solvent-free particles of low vapor pressure solutes. The RESS containing a nonvolatile solute leads to the loss of solvent power by the fast expansion of the supercritical solution through an adequate nozzle, which can cause solute precipitation. The ability to be sprayed is a unique property of supercritical CO2. All the CO2 evaporates and, because the supercritical fluid becomes a gas so quickly in the spraying process, RESS inherently generates nanoparticles. The nozzle configuration plays an important role in RESS method and has a great effect on the size and morphology of the precipitated particles. A co-solvent may be used to improve the solubility of the solute in the solvent. The main features of the rapid expansion apparatus consists of (i) a high pressure pump, (ii) a variable-volume view cell, and (iii) an expansion section. The RESS schematic is shown in Fig. 1.

Figure 1. The schematic diagram of the experimental apparatus for the RESS process

An interesting modification of RESS process is RESSAS (Rapid Expansion of Supercritical Solution in Aqueous Solution). The supercritical solution is expanded through an orifice or tapered nozzle into an aqueous solution containing a stabilizer to minimize particle aggregation during free jet expansion which gives rise to further decrease in the particle size.

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Supercritical Fluids: Simplifying nanotechnology

Megha Swami And Vandana Patravale Institute of Chemical Technology, Mumbai

Email: vb.patravale@ictmumbai.edu.in

Technical Insights

Previously, Turk et al demonstrated and compared the ability for a nonionic polysorbitan ester and polyvinylpyrollidone (PVP K25) to stabilize naproxen particles produced by RESOLV. Results showed average particle size of 300nm for PVP k25 stabilized suspension while expansion in tween 80 shows particle size of 8μm. [1].

Often low solubility of pharmaceutical compounds (such as Griseofulvin) in sc- CO2 results in a low processing rate. To overcome this drawback researchers have used various liquid co-solvents to enhance the solubility. However, only small amounts of liquid co-solvents are suitable for RESS due to the dissolution of particles in the expansion chamber. Therefore, liquid co-solvents such as Methanol, Acetone, or Tetrahydrofuran can be used. A very promising method to overcome the limitation of low solubility is the use of a solid co-solvent [SC] to enhance the low solubility in sc- CO2. Of late, a modification, RESS with solid co-solvent (RESS-SC) has been proposed by Thakur and Gupta. They applied this process for Griseofulvin, Phenytoin, and 2-Aminobenzoic acid by using menthol as the solid co-solvent. In the RESS-SC process, a solid co-solvent is used to enhance the drug solubility in CO2 and avoiding surface-to-surface interaction to other drug particles and therewith hindering particle growth. Later, the solid co-solvent can easily removed from the solute particles by sublimation. As a typical example of the obtained results, the average particle size of Phenytoin produced by RESS-SC is around 120 nm, which is significantly smaller than 200 nm particles obtained from RESS. In addition, the solubility was enhanced approximately 28-fold compared to that of the conventional RESS process without co-solvent. In addition, due to the improved solubility, the phenytoin production rate in RESS-SC is about 400-fold higher than in the RESS process. The solid co-solvent, menthol, was easily removed from the drug nanoparticles by sublimation and lyophilization, which also allowed the dry powdered nanoparticles to be recovered, all in one step. [2]. In the modification of the RESS process described above, both, the solute and the polymer are dissolved in sc-CO2, followed by the rapid expansion of the ternary mixture. Thereby, the sudden depressurization leads to simultaneous co-precipitation of the solutes and formation of composite particles. Debenedetti and co-workers, investigated a model substance, Pyrene, with l-poly(lacticacid) (l-PLA) and Lovastatin with dl-poly(lacticacid) (dl-PLA). In these investigations, the two solutes were dissolved separately and the supercritical solutions were mixed directly before the precipitation unit. These experiments lead to totally different morphology of the precipitated particles: a uniform distribution of Pyrene within l-PLA and crystalline needles of Lovastatin embedded in dl-PLA microspheres.

Recently Signorell et.al [3] has studied the Influence of polydispersity of poly(lactic acid) on particle formation by

RESS, their investigation reveals that the polydispersity of the polymers strongly affects the size but not the shape of the particles. They found larger particles (∼730 nm) for the PLA with high polydispersity than for the PLA with low polydispersity (∼270 nm). In both cases, spherical particles were formed. Moreover, results clearly show that PLA with high polydispersity is less suitable for RESS processing because the low-molecular weight chains are depleted over time and process conditions are thus not constant.

Engineering of Porous Tissue Scaffolds:

Scaffolds are used in tissue engineering as a matrix for the seeding and attachment of human cells. The creation of porosity in three-dimensional (3D) structures of scaffolds plays a critical role in cell proliferation, migration, and differentiation into the specific tissue while secreting extracellular matrix components. These pores are used to transfer nutrients and oxygen and remove wastes produced from the cells. The lack of oxygen and nutrient supply impedes the cell migration more than 500mm from the surface. The physical properties of scaffolds such as porosity and pore interconnectivity can improve mass transfer and have a great impact on the cell adhesion and penetration into the scaffolds to form a new tissue. Various techniques such as electrospinning, freeze drying, and solvent casting/salt leaching have been used to create porosity in scaffolds. However the disadvantages of these techniques include the use of toxic organic solvent, the formation of thin 2D structures, non-homogenous and limited porosity, irregularly shaped pores, and insufficient pore interconnectivity. Gas foaming of polymers using high pressure or sc- CO2 has emerged in recent years as a promising technique that devoid of all the mentioned drawbacks of the earlier methods. Santo et. al. used supercritical CO2 to fabricate hybrid 3D scaffolds of poly (DL-lactic acid) (PDLLA) loaded with chitosan /chondroitin sulphate nanoparticles for biomacromolecule delivery in tissue engineering. In this study, NPs suspended in ethanol was added to PDLLA powder and pressurized with CO2 in a high pressure vessel to 200bar at 358°C. The fabricated composites had porosity and pore interconnectivity of 56% and 39%, respectively, and displayed adequate mechanical properties (compression modulus of 11.3 MPa) for cell adhesion and support. The feasibility of using this scaffold as a multifunctional material was evaluated by the incorporation of a model protein, bovine serum albumin (BSA), either directly into the PDLLA foam or in the NPs that were eventually included in the scaffold. It was reported that this composite could control the release of BSA as a model protein; therefore, the system was a promising candidate for dual protein delivery system for tissue engineering applications. [4] Scientists also are using supercritical fluids and RESS to

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Technical Insights

apply Teflon to vascular stents, a tool used to open arteries in the heart. Typically, the human body rejects the stainless steel stents by growing tissue around the stent. The body recognizes foreign surfaces and builds up tissue to get rid of them and that reclogs the artery. McClain and Taylor, successfully used RESS process to coat the stents with a nanoparticle matrix of polymer and a drug that prevents tissue buildup. By coating the stent with this mixture, serves the dual purpose of surface that is compatible with the body, and also a drug that releases over a long time [5].

Currently our research group is working on similar principle. Studies have been carried out with various biopolymers and lipids using THARSFC instrument and we are successful in obtaining nano scale particles with good

stability. SEPAREX Pharmaceutical Technology is another manufacturer for supercritical fluid instruments who owns its entire development tool, including various supercritical units from laboratory to pilot/industrial scale designed for a wide range of processes pertaining to SCF Technology Platform.

Conclusion

The results presented in this article show that RESS is a very attractive and simple process for the production of submicron and uniform particles with improved dissolution behaviour. Most of the experimental dissolution curves show promising results for bioavailability of poorly water-soluble drugs.

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References: 1. Türk M. , Bolten D. Formation of submicron poorly

water-soluble drugs by rapid expansion of supercritical solution (RESS): Results for Naproxen J. of Supercritical Fluids 2010;55: 778–785.

2. Thakur R. , Gupta R.B. Formation of phenytoin nanoparticles using rapid expansion of supercritical solution with solid co-solvent (RESS-SC) process, Int. J.Pharm. 2006;308: 190–199.

3. Signorell R. Influence of polydispersity of poly(lactic acid) on particle formation by rapid.

expansion of supercritical CO2 solutions. J. of Supercritical Fluids 2010; 51: 376–383.

4. Santo VE, Duarte ARC, Gomes ME, Mano JF, Reis RL: Hybrid 3D structure of poly(D,L-lactic acid) loaded with chitosan/ chondroitin sulfate nanoparticles to be used as carriers for biomacromolecules in tissue engineering. J Supercrit Fluids 2010; 54: 320-327.

5. McClain J., Taylor D. Stents having biodegradable layers. US20100211164A1.

TOIRNAVS NGKLRMEA NMAPARUSH RBNYAXA RYDESDDR URIBOAODN TIASN NCUEIHM SASVNINFTOAIE AZIDDLUCYSA

PharmaScramble

Unscramble each of the clue words

Technical Insights

8

PharmaRebus

Decipher the hidden phrases

AnswerKey

PharmaScramble

NOVARTIS, GLENMARK, SUNPHARMA, RANBAXY, DRREDDYS, AUROBINDO , INTAS,

UNICHEM, SANOFIAVENTIS, ZYDUSCADILA

PharmaDouble

9

Drug-Nutraceutical Interactions: A Revelation

Introduction

The term “nutraceutical” was coined in 1989 by the Foundation for Innovation in Medicine to cover “any substance that may be considered a food or part of a food, and provides medical or health benefits, including the prevention and treatment of disease” in order to distinguish between functional or medical foods and drugs[1]. Under the purview of nutraceuticals, those used for health benefits are most often derived from herbs. Herbal products may contain a single herb or combinations of several different herbs believed to have complementary benefits. According to a recent report by the World Health Organization, herbal medicines have been, and continue to be, used in every country around the world in some capacity with 70–95% of the population in developing countries depending on these traditional medicines for primary care [2]. Estimated at an annual worth of US$ 83 billion in 2008, the global market for traditional medicines is growing exponentially. In particular, North America and Europe have witnessed a dramatic increase in the use of herbal supplements, reports indicating about 15–20% of individuals on prescription medications also use herbal supplements and less than 40% of patients inform their physicians of this use, even if they experience severe side effects—because of the fear of censure or rebuke[3,4].The fact that many physicians themselves are not always aware of the potential for drug-nutraceutical interactions compounds the problem further. Regulations for nutraceuticals are not as stringent as those for traditional medications, as these products are regarded as food products. Thus, manufacturers are spared the cost and effort of pre-market safety and efficacy testing before the release of herbal products and are also exemptedfrom any post-marketing surveillance.

Additionally, consumers generally perceive nutraceuticals as “safe” because of their natural origins. However, many have adverse effects that can sometimes produce life-threatening consequences[5]. Concomitant use of herbs may mimic, augment, or oppose the effect of drugs. Herbal–drug interactions can be characterized as either pharmacodynamic (PD) or pharmacokinetic (PK) in nature.PK interactions can involve enzymes and transporters that are implicated in drug absorption, distribution, or elimination. These are best defined by changes in drug or nutrient parameters (e.g., bioavailability, volume of distribution, clearance). These interactions commonly involve the same transport or metabolic protein, commonly cytochrome P450 (CYP)enzymes, glucuronosyltransferases (UGT), and P-glycoprotein(P-gp), resulting in induction or inhibition activity. These transport and enzyme proteins are widespread but are predominantly found at the intestinal and hepatic sites. Any substance such as a nutraceutical may interact with a drug by bringing about a change in any one or a combination of these systems.As a result, concentration-dependent activity of a therapeutic molecule is altered at the site of action at the drug-receptor level. PD interactions involve the clinical effect of a drug or physiologic effect of a nutrient[6,7]. Further, the interactions of these herbal supplements in combination with drugs have been discussed. 1. Antiepileptics Numerous herbal medicines have effects on the central nervous system and on hepatic metabolism and thus have the potential to interact with some antiepileptic medications. The interactions of a few antiepileptic drugs with commonly used herbal products of Chinese medicine and Indian Ayurveda are given in Tab. 1.

Drug Nutraceutical Interaction Alprazolam Kava Kava Increased sedation, kava might have additive effects with benzodiazepines, given

that they act on the same receptor and on the same areas of the central nervous system with increased GABA receptors[8]

Phenobarbital Gingko Biloba Ginkgo biloba extract (GBE) induced hepatic cytochrome P450 (CYP450) in rats, especially the CYP2B type, thus reducing phenobarbital plasma concentration[9,10]

Phenytoin Shankhapushpi Shankhapushpireducedseizure control and lowered plasma phenytoinlevels by both pharmacokinetic and pharmacodynamics interactions[11]

Phenytoin Gingko Biloba Increased bioavailability of phenytoin, G. biloba has inhibitory effect on CYP2C9 enzyme[12]

Midazolam Echinacea Echinacea selectively modulated the catalytic activity of CYP3A at hepatic and intestinal sites, causing induction in the liver and inhibition in the intestines, thereby decreasing and increasing Midazolam availability at these respective sites[13]

Table 1: Drug-Nutraceutical Interactions for Antiepileptics

Technical Insights

Ankita Pai, Lalit Kagliwal, Rekha Singhal Institute of Chemical technology, Mumbai

Drug Nutraceutical Interaction Hydrochloro-thiazide

Garlic (Allium sativum) Garlic increased the bioavailability and half-life, along with a decrease in clearance and eliminationrate of Hydrochlorothiazide when administered orally, due to enzyme inducing capacity ofgarlic[15]

Felodipine Grapefruit The bioavailability of Felodipine was more than doubled due to the CYP3A4 inhibitory activity of grapefruit juice[16]

Warfarin Danshen (Salvia miltiorrhiza)

Danshentanshinones inhibited CYP1A1, CYP2C6 and CYP2C11-mediated warfarin metabolism, thus prolonging the circulation of warfarin, leading to increased anticoagulant effect[17]

Digoxin St. John’s Wort St. John's wort may reduce efficacy of digoxin and make a patient a nonresponder, whereas increased toxicity may be anticipated after withdrawal of the herb due to induction of P-gptransporter[18]

Nifedipine Ginger PD interactions lead to a synergistic effect on platelet aggregation inhibition of nifedipine[19]

Diltiazem Guggul (Commiphorawightii)

Gugulipid significantly reduced peak plasma concentration and area under curve of diltiazem[20]

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Table 2: Drug-Nutraceutical Interactions for Drugs used in cardiovascular pharmacotherapy

2. Drugs used in cardiovascular pharmacotherapy These include various classes such as α and β- adrenergic blockers, angiotensin converting enzyme(ACE) inhibitors, antiarrhythmic drugs, anticoagulants, antiplatelets and thrombolytics, calcium channel blockers, cardiac glycosides, and diuretics[14]. The interactions of drugs from the above classes with nutraceuticals is given in Tab. 2. 3. Chemotherapeutic Agents The use of nutraceuticals in palliative and curative care for cancer, together with chemotherapy is highly prevalent. As per one study, 37% of patients receiving chemotherapy used herbal remedies concurrently[21]. A few examples of evidence based drug nutraceuticals interactions are given in Tab. 3.

Conclusion

Drug-drug interactions are commonly recognized

occurrences. Drug-nutraceutical interactions are less well appreciated. There is a dearth of well-documented data in this area and there are few studies that have specifically evaluated herb–drugs interactions. It is vital that precise causes for interactions are diligently explored along with their quantitative evaluation. Thus any probable adverse events will be likely to be avoided. However, obstacles in the achievement of this goal include the limited knowledge of healthcare providers and the absence of stringent regulatory restrictions for nutraceuticals. Hence, it is of primary importance that researchers in the field of healthcare direct their efforts towards the identification of various drug-nutraceutical interactions and that a regulatory body is created in order to standardise the manufacture and sale of such nutraceuticals

Drug Nutraceutical Interaction Irinotecam St. John’s Wort Induced expression of the CYP450 CYP3A4 isoform results in a decrease of plasma

levels of the active metabolite by 42%[22] Doxorubicin Black cohosh Black cohosh had a significant sensitizing effect for doxorubicin showing a

steadydecrease in cell survival with increasing black cohosh[23] Tamoxifen Soy isoflavones

(primarily genistein)

The combination synergistically delayed the growth of breast tumor via decreased estrogen level and activity, and down-regulation of EGFR expression[24]

Docetaxel Genistein Genistein, in combination with lower doses ofdocetaxel, resulted in more growth inhibition in PC-3 prostate cancer cells compared with higher doses of docetaxel treatmentalone. Thus lower toxicity of docetaxel could be achievedin combination treatment with better treatment outcome[25]

Table 3: Drug-Nutraceutical Interactions for Chemotherapeutic Agents

Technical Insights

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References

1. DeFelice F. The Foundation for Innovative Medicine. The nutraceuticals initiative: a proposal for economic and regulatory reform, Food Technol. 1992;6:77

2. Robinson M, Zhang X. Traditional Medicines: Global Situation, Issues and Challenges, The World Medicines Situation. 2011.

3. Eisenberg DM. Trends in alternative medicine use in the United States, 1990–1997. JAMA. 1998; 280: 1569–1575

4. Kaufman DW. Recent patterns of medication use in ambulatory adult population of the United States: the Slone survey. JAMA.2002; 287: 337–344

5. Use of Herbal Products and Potential Interactions in Patients With Cardiovascular Diseases. JAm CollCardiol. 2010; 55: 515–25

6. Chavez ML, Jordan MA, Chavez PI. Evidence-based drug–herbal interactions. Life Sciences 2006, 78: 2146–2157

7. Shord SS, Shah K, Lukose A. Drug-Botanical Interactions: A review of the laboratory, animal, and human a for 8 common botanicals. Integrative Cancer Therap. 2009; 8(3): 208-227.

8. Almeida, J. C., and E. W. Grimsley. "Coma from the Health Food Store: Interaction Between Kava and Alprazolam." Ann Inter Med. 1996; 125: 940-941

9. Umegaki K, Saito K, Kubota Y, Sanada H, Yamada K, Shinozuka K. Gingko biloba extract markedly induces pentoxyresorufin O-dealkylase activity in rats. Jap J Pharmacol. 2002; 90: 345-351

10. Kubota, Y., Kobayashi, K., Tanaka, N., Nakamura, K., Kunitomo, M., Shinozuka, K., Kubota, Y., Kunitomo, M., Shinozuka, K. and Umegaki, K. Pretreatment with Ginkgo biloba extract weakens the hypnosis action of phenobarbital and its plasma concentration in rats. J Pharm Pharmacol. 2004; 56: 401–405

11. Dandekar UP, Chandra RS, Sharma AV, Gokhale PC.Analysis of a clinically important interaction between phenytoin and shankhapushpi, an Ayurvedic preparation.J Ethnopharmacol. 1992; 35(3): 285-8

12. Rodda HC, Ciddi V. Herb – drug interaction of noni juice and Ginkgo biloba with phenytoin.Phcog J. 2010; 2(18): 33-42

13. Gorski JC, Huang SM, Pinto A, Hamman MA, Hilligoss JK, Zaheer NA, Desai M, Miller M, Hall SD. The effect of echinacea (Echinacea purpurea root) on cytochrome P450 activity in vivo.ClinPharmacolTher. 2004;75(1):89-100.

14. Zaret B, Moser M, Cohen L (1992) Cardiovascular drugs, Yale University School of Medicine: Heart

Book (283-304). New York:William Morrow & Co

15. Asdaq SMB, Inamdar M.N. The potential for interaction of hydrochlorothiazide with garlic in rats.Chem-Biol Interact. 2009; 181: 472–479

16. Bailey DG, Arnold JM, Bend JR, Tran LT, Spence JD. Grapefruit juice–felodipine interaction: reproducibility and characterization with the extended release drug formulation. Br J Clinpharmacol. 1995; 40: 135–140

17. Wu WWP, Yeung JHK. Inhibition of warfarin hydroxylation by major tanshinones of Danshen (Salvia miltiorrhiza) in the rat in vitro and in vivo. Phytomedicine 2010; 17: 219–226

18. Johne A, Brockmöller J, Bauer S, Maurer A, Langheinrich M, Roots I Pharmacokinetic interaction of digoxin with an herbal extract from St John’s wort. (Hypericumperforatum) Clin Pharmacol Ther. 1999; 66(4): 338-345

19. Young HY, Liao JC, Chang YS, Luo YL, Lu MC, Peng WH. Synergistic Effect of Ginger and Nifedipine on Human Platelet Aggregation: A Study in Hypertensive Patients and Normal Volunteers. The American Journal of Chinese Medicine 2006; 34(4): 545–551

20. Dalvi SS, Nayak VK, Pohujani SM, Desai NK, Kshirsagar NA, Gupta KC. Effect of gugulipid on bioavailability of diltiazem and propranolol. J Assoc Physicians India. 1994; 42(6): 454-5

21. Engdal S, Steinsbekk A, Klepp O, Nilsen OG. Herbal use among cancer patients during palliative or curative chemotherapy treatment in Norway. Support Care Cancer. 2008; 16: 763-769

22. Mathijssen RH, Verweij J, de Bruijn P, et al. Effects of St. John’s wort on irinotecan metabolism. J Natl Cancer Inst. 2002; 94: 1247–9

23. Rockwell S, Liu Y, Higgins S. Alteration of the effects of cancer therapy agents on breast cancer cells by the herbal medicine black cohosh. Breast Cancer Res. Treat.2005; 90: 233–239

24. Mai Z, Blackburn GL, Zhou JR. Soy phytochemicals synergistically enhance the preventive effect of tamoxifen on the growth of estrogen-dependent human breast carcinoma in mice Carcinogenesis 2007; 28(6): 1217–1223

25. Li Y, Kucuk O, Hussain M, Abrams J, Cher ML, Sarkar FH. Antitumor and antimetastatic activities of docetaxel are enhanced by genistein through regulation of osteoprotegerin/receptor activator of nuclear factor-κB (RANK)/RANK ligand/MMP-9 signaling in prostate cancer. Cancer Res. 2006; 66 (9): 4816-4825.

Technical Insights

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CrossWords

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ACROSS 1. Sealable Enclosure 2. HPLC Component 3. Medical Device 6. Biological Shot 7. Densely packed mass 8. Dense Medium 9. Sizing tool 10. Gaseous Colloidal Suspension 11. Nanocylinder

12. Biodegradable constructs 13. Oral Sandwich 14. Biocompatible integrated circuit

DOWN 1. Branched Structure 4. Topical film 5. Site specific implant 8. Dehydrating technique

Infrared (IR) spectroscopy is one of the methodologies for investigating matter that has a very long history in research and applied sciences. Two centuries ago, the astronomer William Herschel discovered the existence of IR in the electromagnetic spectrum for the first time. Since then many advances have been made that provide today’s scientists with a new methodology applicable in astronomy, chemistry, material science, medicine, and many other disciplines.

Although Albert Michelson built his first interferometer in 1881, the application of this technology required almost a century before it revolutionized IR spectroscopy as we know it. The introduction of computers allowed the digitization of the interference information (called an interferogram) followed by fast Fourier transformation (FFT) of the data into the actual IR spectrum. The heart of this technology is an interferometer that traditionally consists of three parts: a beam splitter, a permanently aligned mirror, and a moving mirror. IR is first split into two separate beams, then systematically brought out of phase by increasing the length of one beam path (using a moving mirror) and then recombined back into one beam carrying the interference pattern (Figure.1)

Figure.1 FTIR Block Diagram

The Fourier transformation can then be used to undo this type of interference pattern by transforming it back into the frequency domain of a normal spectrum. Why would anyone go to the trouble of introducing interferometry to IR spectroscopy, and what are the benefits of interferometry over dispersive technology?

FT-IR spectroscopy is much faster and more sensitive (the multiplex advantage — Felgett); it uses all IR energy simultaneously, thereby achieving much lower noise levels (the throughput advantage — Jacquinot), and an internal laser calibrates the interference information, providing very high wavenumber accuracy and reproducibility (the precision advantage — Connes). In addition, no stray light is generated by interferometry, which leads to more accurate quantitative results than is possible with dispersive or current bandpass technologies. This is especially relevant for many aspects of substance identification because modern mathematical methods permit sophisticated database searching and recognition of complex spectroscopic patterns, such as in the case of biological cell identification based on IR spectra.

FT-IR imaging technology

As with all of today’s methodologies, FT-IR spectroscopy is in the process of moving into imaging technology because detector developments (originally available only to the military for missile guidance technology) have improved dramatically during the past decade to a point that mercury–cadmium–telluride IR arrays can be used for spectroscopy. The migration from single-element detectors to focal plane array (FPA) detectors will again change the world of IR spectroscopy. The new dimension in IR spectroscopy allows a new methodology, which has already been called chemical imaging. In the past, a similar explosion in technology allowed.

Chemical images were generated before any FPA technology was available to spectroscopists. However, because only single detectors ere available at that time, mapping of the sample of interest and reassembly of the spectral information into two dimensions was required. This process normally took many hours or days in cases in which larger areas of samples were to be investigated at reasonable spatial resolution (Figure 2). The introduction of FPA detectors altered this time frame by several orders of magnitude and today permits the measurement of 4096 spectra in seconds to minutes (using a 64 3 64 FPA detector, for example). The time advantage is significant compared with the mapping approach, although, not surprisingly, the noise level of a single spectrum of an FPA measurement is usually not as good as that from a single detector measurement.

13

FTIR Imaging In Bio- Medical Technology and Pharmaceutical Industry

Purnima Parkhi Product Specialist - Molecular Spectroscopy Agilent

Technologies, India Email: purnima_parkhi@agilent.com

Measuring Yardstick

However, the challenges of measuring small sample areas through small apertures (diffraction limit) disappear in FPA measurements, because the spatial resolution is given by the effective pixel size of the FPA detector. The limitations of spatial resolution therefore solely depend on the nature of the light used for the analysis. How does FT-IR imaging work? Generally speaking, it is not very different from single detector measurements with a standard interferometer. The only major difference is that, instead of reading the signal of only one detector, 4096 detector elements have to be read out during the spectral acquisition. This process requires a little more time than reading only a single detector and is directly related to the readout capabilities of the FPA detector electronics.The current setup of FT-IR imaging, therefore, requires a step-scanning approach. What this means is that the moving mirror does not move continuously during the data acquisition, as in standard single-detector measurements, but waits for each detector readout to be completed and then moves on to the next position. In this way, 4096 interferograms are collected simultaneously and in a later step transformed into IR spectra. Because every chemical image acquired by FT-IR imaging technology generates a third dimension (a spectrum at each pixel), these data sets are often called image cubes, and the technology to generate them is often referred to as hyperspectral imaging (Figures 2 and 3).

Figure.2 Linear Array Mapping

Figure 3: 2-D Focal Plane Array (FPA) Imaging

FT-IR imaging data evaluation

The intensities of pixels in images taken at a certain wavelength generally reflect the interaction of this specific radiation at a given location with the chemicals in the sample investigated. This is also true for FT-IR imaging because every frame (image) on its own reflects the interaction of IR radiation with chemicals in the

sample. However, an entirely new dimension is added to this very simple approach if one considers that each pixel in the sample area measured contains an entire IR spectrum. Here is where the term “chemical imaging” comes into play. All 4096 spectra (of a 64x 64FPA) can now be spectroscopically evaluated, which means that the analysis can range from single-band intensity plots to complex mathematical approaches such as library searches, cluster analysis, and artificial neural networks. The information generated can then be reassembled into the original image format, providing distribution plots of, for example, ester intensities or spectra similarities based on reference spectra, principal component analysis, or cluster analysis. Chemical information is directly translated back into the image and can then be compared with visible information of the stained or unstained materials. (Figure.4)

Figure4: Infrared Imaging with an Infrared Camera, referred to as a Focal Plane Array (FPA)

FT-IR imaging examples

I] In biomedical technology

Let us consider a challenge of Screening of large tissue sections with a high spatial resolution (5.5 µm pixel size) in a short period of time - analysis of Hippocampus sections from mice to study Alzheimer's disease. To correlate chemical composition and protein secondary structure with tissue morphology, in order to provide insight into the disease process.

Alzheimer's is a progressive and fatal brain disease that is characterized by dementia (deterioration of mental functions). It slowly destroys brain cells causing memory loss and problems with thinking, reasoning and communication. There is no known cure although medications can slow the progress.

Two abnormal structures called plaques and tangles are prime suspects in damaging and killing neurons in the brain. Plaques build up between nerve cells and contain deposits of a protein fragment called beta-amyloid. Tangles are twisted fibers of another protein called tau. The principal goal of the following example is to study an entire hippocampus section to investigate plaques in their respective microenvironment, correlate protein secondary structure with tissue morphology.

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Measuring Yardstick

15

Figure 5: Tissue components spectra obtained from images

Figure 6: Overlay of white matter, Neuropil, Neuron

Figure 7: White matter vs. Grey matter

Changes in CH2 and CH3 peaks in above overlay of the spectra illustrate differences in lipid content between white matter and grey matter. White matter consists mostly of myelinated axons – myelin is composed largely of lipid tissue veined with capillaries. Grey matter consists mostly of neural cell bodies, neuropil, glial cells, and capillaries contains little lipid.

II] In pharmaceutical industry:

In pharmaceutical tablet production, one of the key measurements of product quality is the standard active content uniformity test. This provides a measure of uniformity of the blend from the assay of a number of tablets (typically 10) taken from the tablet press.

The test usually involves dissolving the tablet for HPLC and only the active content is measured. However, the tablet comprises a number of other ingredients and it is known that these ingredients can impact important product properties such as dissolution (a major regulatory concern at the moment), stability, bio-

availability and various process-quality parameters such as hardness. It is also known that some properties can depend on microscopic size and distribution of the ingredients, both actives and excipients.

Modern tablet presses are capable of producing up to 10,000 tablets per minute, yet the uniformity of content measurement described above is the common technique used to provide information on the quality of the blend. Often very little is known about the distribution of ingredients within tablets and this is known to have a significant influence on quality. With costs of poor quality running into billions of dollars worldwide, manufacturing is under scrutiny; increasing pressures to reduce expensive rework costs in production. It is considered that improved understanding of spatial distribution of the ingredients could prove pivotal in providing a better overall understanding of the blending/tabletting process.

In both formulation development areas and the process analytical support functions involved in troubleshooting poor batches it is extremely useful to be able to identify anomalous distributions of ingredients. FTIR imaging has already been successfully employed in some major pharmaceutical companies to provide invaluable information to help solve manufacturing problems. These successes, combined with the opportunities for technology advances, indicate the technique may even have a future as a production control tool.

Tablets can be placed directly on microscope slides, or if powder blends are to be studied, the powders may be poured into special cups and the surface leveled flat prior to analysis.

An example display showing the kind of image which is generated is shown in Figure 8. This shows the total NIR reflectance image of ca. 5 x 5 mm area of an indigestion relief tablet. Here the sample comprises hydroxides of aluminium and magnesium, sucrose and a number of other minor ingredients. At every pixel in the image one can view the NIR spectrum collected at that pixel, as shown in Figure 9.

Figure 8: Total NIR reflectance Image

Measuring Yardstick

16

Component 1 Component 2 Component 3 Figure 10: Typical tablet component images

Figure 11: Single wavelength image: API 5964cm-1

Figure 12: Major component images often complement each other

Figure 9: Spectrum from point 2

Measuring Yardstick

Control Blend

•Control Blend had normal dissolution.

•Poor Blend had slower dissolution.

•Matrix Level difference relates to distribution and particle size of disintegrant within the blend.

Conclusion:

FPA-FTIR provides information on the tissue composition in and around developing plaques. It allows for the in situ study of both the secondary structure of proteins and the functional group content of tissue sections with minimal sample preparation, at the cellular level. The content of the sample was unaltered, providing information that cannot be gained by any other method.

The advantage of chemical imaging in Pharmaceutical

Industry compared to other sensor technologies is the ability to analyse the spatial distribution of the component materials in blends, granules and finished dosage forms. This approach has the potential to monitor

processes to reveal the extent of ingredient blending, particle size distributions, agglomeration of component particles and the presence of polymorphs, hydrates and other trace contaminants”.

FTIR Microscopy and Imaging can help you to solve your complex analytical Problems in Pharmaceutical, Biomedical Technology. Because you See more. See easily. See faster !!!

17

Poor Blend

Figure 13: Comparison between poor and control blend of formulation

Measuring Yardstick

18

This puzzle is a substitution cipher in which one letter stands for another. If you think that Q equals C, it will equal C throughout the puzzle. Single letters, short words and words using an apostrophe give you clues to locating vowels. Solution is by trial and error. In this puzzle U equals to T

PharmaCrypt

Introduction

“The seeds of knowledge may be planted in solitude, but must be cultivated in public.” -Samuel Johnson. The principal objective of patent system is not only to provide benefit of exclusivity to the inventor for a limited period of time and encourage innovation but more importantly it is to promote the development of technology and foster its dissemination such that the invention is accessible to the public at large. In order to strike a balance between the exclusive rights conferred by patents and interests of society at large, patent systems around the world have at least the following features in common (i) patent system must aim to reward only qualified inventions which meet the standards of novelty and inventiveness (ii) invention should be sufficiently disclosed to the public (enablement) (iii) term of protection of an invention should be for a specified period of time (iv) mechanism to prevent abuse. Patent systems around the world comply to a large extent with the above mentioned four features and have been developed with the intention to find an optimal balance among various stakeholders operating in the specific social cultural and economic environment of the concerned country. In the United States (US) patent laws are innovator centric since the stakeholders are innovator companies. On the other hand, Indian approach towards patenting has been driven by the need for providing affordable healthcare to the masses thus enabling a thriving generic industry [1]. As trade barriers diminish and global economies continue to expand, harmonization and enforcement of International patent protection becomes increasingly important. Attempts to balance interests of innovator and generic manufacturers have been part and parcel of legislation in many countries. The diverging standpoint of generics and innovators in India and the United States has been illustrated via two landmark cases Novartis vs Union of India and Novo Nordisk vs Caraco.

Origin of Section 3(d) in India

In the post-independence era owing to changes in

political and economic conditions in the country, India did not choose to abandon patent law as a tool of regulatory policy, but instead redesigned it to suit its national circumstances. Until up to 1970 India employed a legislation which gave protection to product as well as process patents. Indian Pharma hardly had any technological base and largely relied on importing bulk drugs and processing into formulations and this led to foreign multinational dominance and drug prices in India were among the highest in the world [2].This patent regime failed to stimulate innovation. As a result, the patents act 1970 recognized only process patents and not product patents [3].By ignoring product patents, the Indian companies incurred little expenditure on research and development and reverse engineered products of MNC’s and thus it was possible for them to make new drugs available in the country at affordable rates [4]. But developing country generic manufacturers became a threat to the Western pharmaceutical cartels that had dominated the International pharmaceutical industry. The key industry players in the United States felt that there was expropriation of US Intellectual property rights and thus wanted to secure higher patent standards from developing nations and this resulted in the precipitation of TRIPS (Trade related aspects of intellectual property rights) agreement. India therefore was required to bring the patent laws in alignment with standards of TRIPS, which ushered in the product patent regime. In order to be TRIPS compliant and at the same time safeguard the concerns of a developing nation, India introduced Section 3(d) via amendment of Indian patent act in 2005[6]. Novartis Case study G Section 3(d) [8] as amended in 2005 reads as follows livec (Gleevec in US), Novartis brand name drug used to treat chronic myeloid leukemia has been the center of a widely watched patent dispute. Novartis patent application relates to the beta-crystalline form of an already known substance imatinib mesylate. The Indian patent office Chennai rejected the patent claims on the ground that the invention was not patentable under

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Patent law and litigation: A comparative perspective of Indian and American doctrines

Madhulika Vishwanathan, Yagna Praveen Kumarand Mahalaxmi Andheria* Panacea Biotec ltd., GRAND Centre, 72/3 GEN block,

TTC industrial area, Navi Mumbai -400710, India. *E-mail: mahalaxmiandheria@panaceabiotec.com

Abstract: Instead of a theoretical approach, this article attempts to revisit Section 3(d) of Indian patent act and Hatch-Waxman act of the United States with a focus on two currently ongoing cases like Novartis vs. Union of India in Supreme Court of India, and Novo Nordisk vs. Caraco in Supreme Court of United States. This article attempts to strike a balance between diverging standpoints of innovator as well as the generic companies in both the cases.

Measuring Yardstick

Section 3(d) of the Indian patent act [7].Section 3(d) [8] as amended in 2005 reads as follows The mere discovery of a new form of a known substance which does not result in the enhancement of the known efficacy of that substance or the mere discovery of any new property or new use for a known substance or of the mere use of a known process, machine or apparatus unless such known process results in a new product or employs at least one new reactant. Explanation: For the purposes of this clause, salts, esters, ethers, polymorphs, metabolites, pure form, particle size isomers, mixtures of isomers, complexes, combinations and other derivatives of known substance shall be considered to be the same substance, unless they differ significantly in properties with regard to efficacy. Section 3(d) denies patent eligibility to new forms of known molecules (incremental inventions) unless they contribute to higher efficacy of the prior form. Novartis filed two writ petitions declaring that Section 3(d) is not compliant with the TRIPS agreement and challenged the constitutional validity of the section. The main argument of the petitioner (Novartis) was that Section 3(d) confers wide discretionary powers on the Controller as the explanation is devoid of any guidelines. Novartis also argued that increased bioavailability of the beta crystalline form of imatinib meant increased efficacy, entitling it to a patent on beta crystalline form of imatinib mesylate. But Madras High Court clarified efficacy to mean "therapeutic effect in healing a disease."The Indian Patent Appellate Board (IPAB) – where appeals for unsuccessful patent applications are heard subsequently applied this interpretation, and held that the salt form of imatinib mesylate did not meet the test of therapeutic efficacy, and therefore confirmed the rejection of Novartis’s patent application. Discontented with this standard, Novartis has petitioned before the Supreme Court to argue against the interpretation of efficacy by the Madras High Court and IPAB [9].

Origin of Hatch Waxman act in United States

The Drug price competition and patent term restoration act (DPCPTRA) informally known as the Hatch Waxman act introduced in 1984 modified the patent act of 1952to simplify regulatory approval of generic drugs by filing abbreviated new drug applications (ANDA).This act also provides incentives to innovators (by way of patent term extensions to compensate for regulatory delay and other marketing exclusivities) as well as generics (by providing 180 day exclusivity to generic companies that have first to file ANDA’s with challenges against patents listed in the orange book ). In the United states a generic company before launching its product in the market has to file paragraph certifications against each and every patent listed in the orange book against the innovator’s product .This certification must state one of the following: Paragraph I certifies that the required patent information has not been filed; Paragraph II certifies that such patent has expired; Paragraph III certifies that the patent will expire on a particular date and the generic

will launch after the expiry of such patent; or Paragraph IV certifies that such patent is invalid or will not be infringed by the drug, for which approval is being sought. In most paragraph IV cases the generic applicant is sued by the patent holder and this begins a process in which the question of whether the listed patent is valid or will be infringed by the proposed generic product may be answered by the courts [10].This event of lawsuit by the innovator triggers automatic 30 month stay on generic approval. The Hatch Waxman act was susceptible to further monopolistic abuses and anticompetitive practices [11] which consequently led to further delays in the release of generic drugs, and significant increases in prescription drug prices. This eventually led to the introduction of Medicare Modernization act (MMA) in 2003. Novo Nordisk vs Caraco pharma The case involves Novo Nordisk's drug repaglinide marketed as Prandin®. Novo listed two patents in the Orange Book associated with this drug: Reissue Patent No.RE37035, which claims repaglinide drug product itself and expired on March 14, 2009. The other patent, US6677358, claims the method of use of repaglinide in combination with metformin; this patent expires June 12, 2018. Apart from this there were two other approved uses for Prandin®: (i) as monotherapy (ii) in combination with thiazolidinediones neither of these indications is claimed in any additional Orange Book listed patent. On February 9, 2005Caraco filed an ANDA for generic repaglinide having a Paragraph III certification regarding the '035 patent and a Paragraph IV certification for the '358 patent, the latter leading to ANDA litigation. During the litigation, Caraco stipulated that since it did not include a label describing the combination (repaglinide and metformin), its ANDA would not infringe the '358 patent. In November 2007, as a part of ongoing revaluation of professional labeling of all oral anti-diabetic drugs, the FDA required Novo to replace all separate indications of Prandin® with the following indication “Prandin is indicated as an adjunct to diet and exercise to improve glycemic control in adults with type 2 diabetes mellitus.”In April 2008, Caraco amended its ANDA to include a split certification by which it maintained its Paragraph IV certification on the non-method as claimed in claims 1-3 and 5 but included a sec viii statement specifying that it did not seek approval to market the generic repaglinide and metformin combination as claimed in claim 4. The "carve-out" label was acceptable to Food and Drug Administration(FDA). After FDA approved Caraco’s sec viii carve-out and shortly after expiry of ‘035 patenton May 6, 2009,Novo filed U-968 code (a method for improving glycemic control in adults with Type II diabetes mellitus) as a significantly broadened replacement for U-546 (use of repaglinide in combination with metformin to lower blood glucose) it had previously submitted for the ‘358 patent. The FDA did not direct or request Novo to change its use code to reflect the new indication nor was Novo

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Measuring Yardstick

required under FDA regulations to make such a change. The ‘358 patent does not cover monotherapy of repaglinide; it covers only repaglinide in combination with metformin. Thus the new use code narrative U-968 submitted by Novo is misleading and incorrectly suggests that ‘358 patent covers three different FDA approved uses for repaglinide viz. (i) monotherapy (ii) combination therapy with thiazolidinediones (iii) combination therapy with metformin [12].This change in the use code caused the FDA to reject Caraco's Section viii certification and "carve-out" label, requiring Caraco to amend its label as per the directive as issued for Prandin in November 2007.Since Caraco stipulated that this amendment would amount to an infringement, Caraco counterclaimed in district court for an injunction requiring Novo to return the use code to U-546. The District Court granted injunction on this issue, and ordered Novo to change the use code in the orange book from U-968 to its former U-546 listing. Novo appealed this district court order in the federal circuit. The Federal Circuit held that the statute has no provisions permitting a generic drug maker to obtain an order from a court (like the injunction here) to compel a patent holder to change or modify its use code. Thus the Federal circuit reversed the decision of district court and vacated the injunction [13]. If the innovator submits a use code narrative that is broader than its patent claims such that it also encompasses unpatented uses it could effectively block the generic manufacturer’s use of the carve out provisions forcing the generic to either wait for the expiration of patent or force the generic to litigate a patent it never intended to infringe. The statutory interpretation of the counterclaim provision of the Hatch Waxman i.e. whether a generic company can sue an innovator to get it to narrow its use code narrative is now with the Supreme Court.

Discussion

The Supreme Court’s verdict on Section 3(d) might clear the ambiguity associated with this section. Although Section 3(d) prohibits evergreening by denying patents to incremental inventions, it raises many questions whether the “regulatory body” or “patent office” should determine the standard of proof regarding efficacy. While it is incumbent upon society to respect the intellectual labor of the inventors, removal of section 3(d) thereby paving way for evergreening of existing patents

in pharmaceuticals will be counter productive to affordability and accessibility of healthcare in India. In United States the complex regulatory framework of the Hatch-Waxman act and Medicare Modernization act is designed to strike a delicate balance between innovation and competition in the pharmaceutical industry. The Supreme Court’s decision in Caraco v. Novo Nordisk will have an important impact on that balance by either permitting patent holders to expand patent use code narratives in a manner that could impede approval of generic drug applications, or giving generic manufacturers a means by which to challenge the scope of patent use code narratives by using counterclaim provisions as provided in MMA 2003.

Conclusion

Such complex patent laws and litigations and its cost and time implications, opens the door for researchers to explore in the field of new drug delivery systems and patenting them. This could potentially stretch the lifecycle of a molecule which has already enjoyed its patent term via product patent and use patents etc. This strategy not only encourages researchers to incline more towards innovative research programs, but also results in significantly improved, efficacious, well tolerated and safe medications as compared with their conventional counterpart dosage forms and thereby benefiting the patients. One such example being commercially successful product ABRAXANE®, (PacliALLTM of Panacea Biotec Ltd) for treating cancer. This is a nanoparticulate injection based on NABTM technology. The active agent in ABRAXANE® is paclitaxel which is a chemotherapeutic agent known since 1967. Paclitaxel was initially formulated as TAXOL® by Bristol Meyer Squibb using Cremophor ELTM and ethanol as solubilizer. Owing to serious hypersensitivity reactions associated with TAXOL®, Abraxis Biosciences (Now Celgene) had developed a NAB based Paclitaxel injection which has eliminated hypersensitivity reactions associated with TAXOL®. By doing so, Abraxis Bioscience not only ensured safer and well tolerated medication but also provided a very well-guarded patent protection till at least 2013 in US. By this way a product which is known since 1967 has been protected till at least 2013.

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Measuring Yardstick

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References:

1. Chandran, Sanjeev et al; Implications of New Patent Regime on Indian Pharmaceutical Industry: challenges and opportunities; Journal of Intellectual property rights 10(4):269-280 (2005)

2. B.K.Keayla; Amended Patents Act: A critique; Combat Law, 4(2); 2005

3. The patents act 1970; http://wbbb.gov.in/Legislations/rules/TheIndianPatentAct1970.pdf (last accessed 1/23/2012)

4. Andrade Chittaranjan et al; The new patent regime: Implications for patients in India. Indian Journal of Psychiatry 49(1) :56-59 (2007)

5. Peter Drahos, Global Property Rights in Information: The Story of TRIPS at the GATT' Prometheus 13(1) :6-18 (1995)

6. The patents (amendment) act 2005; http://ipindia.nic.in/ipr/patent/patent_2005.pdf (last accessed 1/23/2012)

7. Thomas Zakir ;IP case law developments;Journal of Intellectual property rights 12(1): 507-515 (2007)

8. Before the amendment in 2005 Section 3(d) read as: the mere discovery of a new property or new use of

a known substance or new use of a known substance or of the new use of a known process ,machine or apparatus unless such known process results in a new product or employ at least one new reactant

9. What future for India’s Patent Act? Novartis vs. Union of India

http://www.msf.org.za/publication/what-future-india%E2%80%99s-patent-act-novartis-vs-union-india(last accessed 1/23/2012)

10. Implementation of Provisions of the Drug Price Competition and Patent Term Restoration Act of 1984;http://www.fda.gov/newsevents/testimony/ucm115218.htm(last accessed 1/23/2012)

11. American Bar Association section of antitrust law ; Antitrust counterattack in intellectual property- litigation handbook (ABA publishers 2010)

12. United States District court eastern division of Michiganhttp://www.fdalawblog.net/files/prandin---9-24-use-code-decision.pdf (last accessed 1/23/2012)

13. United States Court of Appeals for Federal circuit http://www.fdalawblog.net/files/prandin---apotex-amicus-brief.pdf (last accessed 1/23/2012)

Measuring Yardstick

The editorial team of CRS Newsletter invited me to

share the experience of journey of “Fungisome - from Bench to Bedside” to get insight for finding ways to improve commercialization possibilities of Nanotechnology based products under development at both academic and industry research centres in the country. While it is not necessary that all initiatives need to tread the path trodden by Fungisome, yet a success story would always have some guiding facets and thus sharing the lessons learnt could serve as a useful guide.

The launch of FUNGISOME i.v. - a completely indigenously developed superior alternative to all imported Lipid and Liposomal formulations of Amphotericin B on May 11, 2003 by Lifecare Innovations coincided with Technology Day – a day dedicated to India’s technological prowess. The launch of FUNGISOME i.v. kindled hope in the Indian Pharma industry and marked the beginning of moving beyond generics. FUNGISOME i.v. is the first Liposomal Drug from Asia, and being a Nanosomal preparation it is also regarded to be the first Nano-Drug of India. It has been acknowledged to be the most nephro-safe Amphotericin B formulation of the world. FUNGISOME i.v. most effectively addressed a critical half-a-century old problem of Amphotericin B toxicity. The reason for the unprecedented success of FUNGISOME i.v. is that it is indeed an innovation with distinct advantages over the Gold Standard of the time, viz AmBisome with respect to safety/toxicity, efficacy, dose and resultant matchless clinical outcome and economy of systemic and invasive mycosis treatment.

Many initiatives on development of Liposomal and Nano-Drugs, both within the industry and at academic institutions followed soon after. Lifecare Innovations has over the years commercially launched four more Lipid/ Liposomal novel formulations.

The development of FUNGISOME i.v. started not for expanding product portfolio of or revenue generation for a pharma company. In fact, a number of companies from mid 1990s to early 2000s that I had reasons to interact with, lacked vision and resisted engaging in Liposomal Amphotericin B. None was ready to risk any investment, howsoever small. Unfortunately most minds in the industry seem to look for easier ways and tend to be engrossed with “Process Development” a euphemistic way to skirt patents. It was, however, a critical medical need to develop a safe, effective and affordable drug to treat systemic mycosis and leishmaniasis (Kala-Azar). FUNGISOME i.v. fitted the bill, since the demand was

great and supply non-existent.

Almost a decade after launch of FUNGISOME i.v., the outcome of the Indian R&D on “Innovations in Nanotechnology based Pharmaceuticals” is dismal and compels us to ponder. The potentials of commercial success can be ascertained at the early stages of development.

NDDS based pharmaceuticals that include Nano-Drugs and Liposomal Drugs, have a distinct feature that they do not adhere to concept of generics. In addition to APIs their carrier particles in these NDDS, also contribute to drug action predominantly mediated through strategically designed delivery and controlled release patterns. The carrier nano-particles of NDDS are required to be designed objectively. Not only the composition but the process of their assembly is critical. It is an extensively proven and documented fact that nano-carriers of identical composition exhibited different toxicity and efficacy. Conventionally, identical compositions qualify to be granted as Generics. However, in case of NDDS, the products need to be subjected to pre-clinical toxicology and clinical trials to establish that the re-production is indeed a generic, innovatively improved or mere imitation. If the clinical performance of a follow on product is sub-par to the original product it becomes an imitation and not a generic. For the purpose of brevity, an excerpt from an earlier presentation makes interesting reading.

Innovation has many definitions, and the one relevant in the present context of nanotechnology based NDDS drugs is – “addressing an unmet need through the novel applications of ideas and technologies”. An imitation of such an innovation shall not and cannot be misconstrued to be also an innovation. Before undertaking the journey to develop a drug, it would be prudent to determine if the outcome would be an innovation or imitation. One must question self as to “what problem the aspired innovation would solve”. One must not attempt to solve a problem that does not exist. An ideal innovation is the one that doctors and patients are waiting for. No one waits for an imitation. Commercial success would remain a mirage if imitation is aspired to go commercial in the disguise of innovation. Good Science is an essential requisite for commerce in the area of modern medicine.

In my two decades of involvement with the Indian industry, I have failed to notice any distinct enthusiasm for innovation. A majority of the industry seems to be driven by quick buck approach and going global with

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Business Snapshot

Commercialization – a Catalyst for Drug Discovery and Innovation

Jitendra N Verma, PhD Managing Director, Lifecare Innovations Pvt Ltd.

Gurgaon - 122009 (INDIA) Email: jnverma@lifecareinnovations.com

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CONCEPT OF GENERICS OF CONTROLLED RELEASE PHARMACEUTICALS (NDDS) IS SCIENTIFICALLY UNFOUNDED: Liposomal AMPHOTERICIN B AN EXAMPLE.

Abstracts- 11th International Symposium on Advances in Technology And Business Potential of New Drug Delivery. Controlled Release Society – Indian Chapter.

Mumbai (February 2011) Controlled Release Pharmaceutical formulations including Liposomes, Lipid Complexes & Particles in Colloidal Dispersion, Nanoparticles and Microparticles constitute a promising range of Novel Drug Delivery Systems. Constituent molecules viz. phospholipids/ lipids and biodegradable polymer/s which form the matrix of the carrier particles in these formulations impart tremendous flexibility because of the large range of molecules to choose from. Chemical characteristics and composition of carrier particle constituents, their ratios, the physico-chemical properties of the drug incorporated and the process for the assembly of these particles results in uniquely different shapes, sizes, lamellarity, charge, encapsulation efficiency and stability of carrier particles in each preparation which impart it's distinct in vivo behaviour including tissue distribution pattern, interaction with cells, drug targeting pattern, drug release profile, dose/range and pharmaco-kinetics etc. which together result in different therapeutic index of each preparation. Number of NDDS formulations of Paclitaxel, Doxorubicin and Amphotericin B are commercially available and at different stages of development but they are not identical in their clinical outcome. Similar to Phosome, Lambin and Ambilip in India, an Argentinian product Anfogen*- generic of Ambisome a Liposomal Amphotericin B is reported to be ten times nephrotoxic and much less effective than Ambisome. Other formulations of Amphotericin B viz, Amfy and Amphomul have no known comparable lipid or liposomal Amphotericin B yet claimed be safer despite reports that Amphomul is as nephrotoxic as conventional Amphotericin B Deoxycholate suspension. Fungisome, Ambisome, Amphotec/Amphocil and Abelcet/Ampholip, are documented to have successful response of 91%, 77%, 46% and 34%.; nephrotoxicity of 1.8%, 10-20%, 25-40% and 42-63%; and dose 1-3mg/kg, 3-5mg/kg, 5mg/kg and 4-6mg/kg, respectively. Such observations warrant that all NDDS even if same in chemical composition be treated as new formulations and be subjected to clinical trials for ascertaining safety and efficacy and not pose threat to patients. Treating NDDS as generics defeats the very objective of improving performance of known therapeutics. *Comparison of the Pbysicochemical, Antifungal, and Toxic Properties of Two Liposomal Amphotericin B Products., Jon A. Olson,1 Jill P. Adler-Moore,l,5* Gerard M. Jensen,2 Julie Schwartz,3 , Cecilia Dignani,4 and Richard T. Proffitt, ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, Jan. 2008, p. 259-268 Vol. 52, No. 1

Business Snapshot

their generics. Academia has its own limitations to plan research around their Ph.D. aspirants, their need to publish papers and move westwards to brighten their career. New, novel, innovative products are rarely a priority of industry or academic institution. The entire country continues to show tremendous dependence on IP from outside the country.

India needs an enabling ecosystem for drug discovery and development programs. Ideal programs are built on strong foundation of basic research that includes parallel development of laboratory methods and subsequent scale-up of processes to manufacture, development of parameters to ascertain and assure quality, along with the involvement of government for education of regulators, doctors, medical educators and procurement agencies.

Having identified the product and specifics, the immediate and next priority to comprehend is IP issues. A development without patentability would be of little value to the industry. Both, the patentability and the funds required for IP protection are bottlenecks for most academic labs. Ironically most public funding for IP protection is limited to filing Indian Patent which amounts to disclosure without any meaningful protection. The end result is that such initiatives add only to nothing more than statistics of successfully completed R&D projects.

IP protection significantly raises the possibility of developments needed for commercialization. At this point fulfilment of regulatory compliances takes centre stage. Though in the past regulatory system specifically for NDDS had been archaic, it is a good sign that the present team India of regulatory and research funding agencies is apparently gearing up to provide regulatory guidance and minimize bureaucratic impediments to optimize commercialization.

FUNGISOME i.v. was aptly celebrated as “India Succeeds”. It was not only the innovators and entrepreneurs, but also DBT, DSIR, NRDC, and DCGI that lent all the fiscal support and accorded timely approvals.

But for their “Spirit of Team India”, FUNGISOME i.v.

would have been yet another file, gathering dust in various offices. It is in this context that the need for networking with the industry, regulators, technical experts, funding agencies at early stages of innovative initiatives are essential in the journey from “Mind to Market”.

The timing of filing patent requires a great deal of prudence. Patent life is limited irrespective of whether the product goes commercial or not. Therefore, funding for pre-clinical studies and clinical trials needs to be organized early on. The cost of pre-clinical and clinical trials together itself is multi-million US$. Both the academic labs and start-ups, micro, small and medium enterprises (MSMEs) need 100% funding to move forward the product in developmental pipeline. Today the partial grants provided through various government schemes is discouraging. It is also important to receive funding without delays caused by bureaucratic procedures as the procedural delays would eat up the patent life. Discretion for granting funds and their disbursal should therefore be vested in the technical committees. The country must simplify procedures in the interest of development and commercialization of the products required for health and longevity. Although I appreciate the enthusiasm of technocrats in all the departments of the government, but I often see them working with their hands tied behind their back. If our fiscal policies are not goal oriented, we will have policies that would not deliver products. At the same time it is also not only in his interest but also incumbent on the innovator and entrepreneur to be participative in the framing of progressive policies while keeping requisite accountability in place.

A rightly identified NDDS that has strength of science, is IP protected, has adequate funding resources for pre-clinical/ toxicology and clinical trials would be considered to have potential for commercialization and would draw attention of all stake holders. Nothing succeeds like success and one success would generally catalyze another success.

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• Prof. Padma V. Devarajan, Head, Dept. pharm sci. tech. Institute of Chemical Technology, Mumbai, India received AAiPS DISTINGUISHED EDUCATOR AND RESEARCHER AWARD – 2011 by American Association of Indian Pharmaceutical Scientists (AAiPS) at 2011 Annual Meeting and Exposition of the American Association of Pharmaceutical Scientists (AAPS) held at Washington D.C, USA from 23-27 Oct 2011. and was awarded 2008 VASVIK Smt. Chandaben Mohanbhai Patel industrial research award for women scientists for technological contributions to nanomedicine by vividhlaxi audyogik samshodhan vikas kendra, october 7, 2011. She also featured as Indian Women Scientist in Chemical Industry News, June 2011 a monthly journal published by the Indian Chemical Council, as a recognition for contributions to research in science in technology in a special issue celebrating the International Year of Chemistry - Contributions of Indian women scientists

• Prof. V. B. Patravale, Professor of Pharmaceutical, Institute of Chemical thecnology, Mumbai, India received Fellow of Maharashtra Academy of Sciences award, Maharashtra Academy of sciences.

• Dr. Prajakta Dandekar-Jain, featured an article “Vigyanvati” under the category of Youth Icon, Maharashtra Times, August 29, 2011.

• Anisha D'Souza received partial travel support by INSA to for attending and presenting poster at the 2011 AAPS

Annual Meeting and Exposition, October, 23- October, 28 – August 3, 2011 in Washington. • Bharti Goswami received travel Grant from Controlled Release Society Indian Chapter to attend and give Poster

presentation in the 38th Annual Meeting and Exposition of the Controlled Release Society, Maryland, July 30-August 3,2011.

• Praveen Date Travel grant award by ICMR to for attending and presenting poster at the 38th Annual Meeting & Exposition of the Controlled Release Society, July 30 – August 3, 2011 in National Harbor, Maryland.

• Dr. (Mrs.) M. S. Nagarsenker received travel award from Indian Council of Medical Research (ICMR) and Gattefosse India Pvt. Ltd for attending and presentation of poster entitled “Compritol® 888ATO a release modifier for sustained release of highly water soluble agent: Formulation, Evaluation and IVIVC study” at 38th Annual Meeting & Exposition of the Controlled Release Society held at National Harbor, Maryland, USA, July-Aug 2011

• Shilpa Patere received travel award from Department of Biotechnology (DBT), Indian Pharmaceutical Association (IPA) and Gattefosse India Pvt. Ltd. for attending and presentation of work entitled” ; Liposomal Drug Delivery System for Receptor Based Hepatic Targeting” at 38th Annual Meeting & Exposition of the Controlled Release Society held at National Harbor, Maryland, USA, July-Aug 2011

• Sandhya Pranatharthiharan was receipient for travel grant award by Abitec Corporation to for attending and presenting poster at the 38th Annual Meeting & Exposition of the Controlled Release Society, July 30 – August 3, 2011 in National Harbor, Maryland.Prof. Padma Devarajan was receipient of travel grants from DST, Centre for International Cooperation in Science(CICS) and ICT Golden Jubilee/ NOCIL grant for attending and presenting poster at the 38th Annual Meeting & Exposition of the Controlled Release Society, July 30 – August 3, 2011 in National Harbor, Maryland.

• Pratik Patel received travel grant from DBT, India, to attend and present poster at 9th International Conference and Workshop on Biological Barriers in vitro and in silico Tools for Drug Delivery and Nanosafety Research, February Department of Biopharmaceutics and Pharmaceutical Technology of the Saarland University Campus Saarbrcken, Saarbrcken, Germany 2012.

• Ramchandra Patale received travel grant from DBT, India to attend and present poster at 25th Annual Meeting & Exposition of the American Association of Pharmaceutical Scientist (AAPS), October 23 – 27, at Walter E. Washington Convention Center, Washington, DC, USA

• Sanket M. Shah received travel award from ICMR-BMBF for visiting and working at Pharm Tech Dept, F.S. University of Jena, Germany for a period two months from 1st Dec. 2011 to 1st Feb. 2012

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Hall of Fame

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• Dr. Prajakta Dandekar-Jain appointed as Dr. John Kapoor Assistant Professor of Pharmaceutical Technology at Institute of Chemical Technology, Matunga, Mumbai, 2012.

• Dhami Manju & Prof. K. K. Singh received Second Prize for the research work “Brahmi Ghrita and Extract in a novel improved form for memory enhancement ” 5th Interuniversity research festival of “Avishkar 2011” on January 13-14, 2012 held at Shivaji University, Kolhapur, Maharashtra

• Dr. Ratnesh Jain received Ramanujan Fellowship from Department of Science and Technology, Govt. Of India, at Institute of Chemical Technology, Matunga, Mumbai, 2012.

• Nirale NM, Kshatriya A, Shirole R, Saraf MN, Nagarsenker MS, Kshirsagar NA. Best poster presentation award for presenting the research work entitled “Beclomethasone Dipropionate- Dry Powder Inhalation” at 5th International conference on Drug discovery and Development-South Asian perspective organized by South Asian chapter of American College of Clinical Pharmacology held in Mumbai December 2011.

• Pathak PO, Nagarsenker MS, Barhate CR, Padhye SG, Viswanathan CL, Bhattacharyya B, Ghosh SS and Chaudhari PR. Best oral presentation award for presenting the research work entitled “Novel Polysaccharide Guided Liposomal System for Targeting Hepatocellular Carcinoma” at 5th International conference on Drug discovery and Development-South Asian perspective organized by South Asian chapter of American College of Clinical Pharmacology held in Mumbai December 2011.

• Pandharipande P. P., Dalapathi G. B., Desai P.P., Patravale V. B. ‘Freeze Drying : Exploring Potential In Development Of Orodispersible Tablets Of Sumatriptan Succinate’ Best poster award at 63rd Indian Pharmaceutical Congress, Bangaluru, India, 16-18th December 2011.

• Vivek Borhade and Harshad Shete, Vandana Patravale. “Atovaquone Nanosuspension For Intravenous Delivery: Toxicity Assessment, Pharmacokinetics, Tissuedistribution And In Vivo Antimalarial Efficacy Studies” Best paper award at INDO-US Joint Symposium on Nanomedicine : Prospects and Challenges, Mumbai, India, 14-15th November 2011

• Rohan Pai- “Chitosan – based microparticles for inhalation delivery of Rifampicin and Rifabutin .” At Nasal & Pulmonary Seminar – Nov. 2011, Mumbai organized by IPA This was adjudged as second best poster.

• Shruti Hazare-" Evaluation of 99mTc-microspheres for lung deposition studies using gamma camera". This was awarded 3rd Prize At 43rdAnnual Conference of Society of Nuclear Medicine, India at Chennai, Dec. 2011.

• Medha Patel & Prof. K. K. Singh awarded Second Best Poster Award for the poster entitled“Herbal Nanomedicine: A Paradigm for Effective treatment of Psoriasis” at the Second Word Conference on “Nanomedicine and Drug Delivery” WCN - 2011 on 11th-13th March, 2011held at Kottayam, Kerala

• Soni U., Desai P., Pathak S., Sharma S., Patravale V.B. “Improved therapeutic efficacy with nanosized halofantrine: potential of dose reduction” Award for poster at Eleventh International Symposium on Advances in Technology and Business Potential of New Drug Delivery Systems, Mumbai, India, 16-17th February 2011

• Soni U., Pathak S., Sharma S., Patravale V.B. “Nanoemulsion of halofantrine: rediscovering the potential of banned drugs” Award for poster at Eleventh International Symposium on Advances in Technology and Business Potential of New Drug Delivery Systems, Mumbai, India, 16-17th February 2011 .

• Dhaivat C. Parikh, Tejal A. Mehta, Avani F. Amin received "Merit Award" for Research Poster on “Formulation Development of Gastroretentive Drug Delivery System For Cinnarizine : In Vitro and In Vivo Investigations” at the 11th International Symposium of CRS-Indian Chapter on “Advances in Technology and Business Potential of New Drug Delivery Systems” held in Mumbai on 16th & 17th February 2011.

• Ranjita Shegokar, Bharti Goswami & Prof. K. K. Singh awarded First Prize for the poster entitled “NEVARAPINE NANOSUSPENSIONS: BIODISTRIBUTION STUDIES AND TARGETING TO HIV RESERVOIRS”at the 11th International Symposium on Advances in Technology and Business Potential of New Drug Delivery Systems entitled at the 11th International Symposium on Advances in Technology and Business Potential of New Drug Delivery Systems organized by Controlled Release Society-Indian Chapter on 16th -17th February, 2011 held at Mumbai.

• Pallavi Pople & Prof. K. K. Singh awarded Certificate of Merit for the poster entitled “Modified Nanolipid Carrier: Novel Strategy for Higher Entrapment and Improved Performance for Topical Delivery of Tacrolimus” at the 11th International Symposium on Advances in Technology and Business Potential of New Drug Delivery Systems organized by Controlled Release Society-Indian Chapter on 16th -17th February, 2011 held at Mumbai.

• Medha Patel & Prof. K. K. Singh received Certificate of Merit Systems for the poster entitled “Nanoflax: Herbal Nanocarrier System For Targeted Delivery and Effective Treatment of Psoriasis” at the 11th International Symposium on Advances in Technology and Business Potential of New Drug Delivery Systems organized by Controlled Release Society-Indian Chapter on 16th -17th February, 2011 held at Mumbai.

Awarding Excellence

CRS News

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