Project Evaluation Report

1. Project Title: Advanced Drug Delivery Systems (ADD)

2. Project Details:

Nodal Lab CSIR-IICT

Participating CSIR-CDRI, CSIR-NCL, CSIR-IGIB, CSIR-CLRI, CSIR-IITR & non CSIR-Labs Labs such as lACS, Kolkata; liSe. Bangalore; KEM Hospital, Mumbai

Nodal Officer Dr. Arabinda Chaudhuri

Taskforce Director, CSIR-IICT Chairman

Total Man Days 7 484 (25 Scientists)

Approved Cost Rs 1557.91 Lakhs

Fund received Rs 1286.409 Lakhs

Fund utilized Rs 1175.506 (until Oct 2016)

3. Periodic Key Recommendations of Task Force/RC/Monitoring Committee

S.No Key Action taken on Proposed Deliverables

Recommendations Key of Task Recommendations Force/RC/Monitoring Committee

1. To focus on in vivo Performed as studies with leads recommended available

2. Suggested to expand In vivo studies in vivo studies in expanded to skin, multiple tumor models pancreatic, colon,

brain and lung cancers

3. Task force committee Talks and recommended to look negotiations out for potential initiated with buyers for at least 3 multiple companies technologies from India and

Revised Deliverable (approved by Task Force)

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I developed in IICT. 1 abroad .

4. Project Achievements: [Highlighting only those studies that showed successful preclinical outcome]

S. No. Deliverables Deliverables Achieved If not promised at start achieved,

reasons thereof

WP1 Development of 1. Novelliposomal formulation of (CSIR- targeted lipid, essentially water insoluble anticancer IICT) polymer, dendrimer- drug curcumin (-500 ~g/ml) has been

and inorganic developed using the newly synthesized nanoparticle-based RGDK-Iipopeptide containing guanidine delivery system head-groups and two stearyl chains

(CSIR-IICT). It has been demonstrated that such curcumin formulation can significantly inhibit growth of solid melanoma tumor through targeting curcumin to mouse tumor vasculature (Mondal, G. et al. J. Control. Release.2013, 172, 832-840; IF= 7.4) .

2. An efficient liposomal formulation of circulation stable pegylated RGDK-lipopeptide has been developed (by CSIR-IICT) for inhibiting melanoma growth in mice by delivering liposomally encapsulated anticancer CDC20siRNA to tumor vasculature ( Majumder, P. et al. J. Control. Release.2014, 180, 1 00-108; IF= 7.4) .

3. An efficient circulation stable liposomal formulation of pegylated RGDK-lipopeptide has been developed (by CSIR-IICT) for simultaneous delivery of both hydrophobic & hydrophilic (curcumin & doxorubicin, respectively) to mouse tumor vasculature (Barui, S. et ai.Biomaterials2014, 35, 1643-1656; IF = 8.55) .

4. CSIR-IICT scientists have demonstrated that CDC20siRNA and paclitaxel co-loaded nanometric

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liposomes of nipecotic acid-derived cationic amphiphile inhibit xenograftedneuroblastoma (Nanoscale 2017, 9, 1201-1212; IF= 7.78).

5. An efficient direct in vivo dendritic cell targeting liposomal DNA vaccine carrier (containing cationic lipids with mannose mimicking shikimoyl or quinoyl head­group) has been developed by CSIR­IICT (Garu, A. et al. Mol. Ther. 2016, 24, 385-397; IF= 6.93; WO 2014106856 A1 (Publication date: July 10, 2014; PCTIIN20131000806 filed on 27/12/2013; Indian Patent Application No. 0017/DEL/2013, filed on January 3, 2013).

6. A novel combination approach (a potentially transformative platform technology) for eradicating/regressing established tumor has been developed by CSIR-IICT (Indian Patent Application No. 2442/DEU2013 filed on19/8/2013; US Patent Application No. 14/462880 filed on 19/08/2014. Manuscript under preparation.

New GR-targeted lipid and AuNPs based drug & gene delivery reagents against drug resistant cancer cells and pancreatic cancer cells have been developed by CSIR-IICT (Pore S, et ai.Biomaterials2013, 34, 6804-6817 (IF = 8.55); Mondal, S. K. et al. Mol Pharm 2016, 13, 2507-2523; Ahmad, A. et al. Mol Pharm 2016, 13, 1081-1088 (IF = 4.5).

In collaboration with CSIR-CCMB, CSIR­IICT scientists have developed an engineered fusion protein-loaded gold nanocarriers for targeted co-delivery of doxorubicin & erbB2 siRNA to HER2+ ovarian cancer has been developed (Patent Application number 201611036499 dated 25.10.2016).

Tumor targeting inorganic nanoparticles for combating cancer have been

Funds for sanctioned confocal microscopy was not obtained in 1-4th year. The microscope was finally procured in 5th year. The originally proposed work has started. However, the revised work plan worked using

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WP2 (CSIR­CDRI)

WP3 (CSIR­NCL)

GR-based cancer detection kit Targeted drug delivery methods for cancer

Development of glycopeptide, lipid, hyaluronic acid, lectin, biological nanoparticle-based drug delivery systems

developed by CSIR-IICT (Patra and co- fluorescent workers, Theranostics2014, 4, 316-335 inverted (IF= 8.4). microscope.

Most recently, CSIR-IICT scientists have demonstrated that lipocardiaromidewith C12 chain length possessesintrinsic anticancer activity and can deliver therapeutically important siRNA to tumor cells (patent filed)

Not achieved as proposed. However, partially achieved by the development of other classes of molecules exhibiting selectivity towards cancer cells.

1. CSIR-CDRI scientists have demonstrated enhanced apoptotic and anticancer potential of paclitaxel loaded biodegradable nanoparticles (Gupta, U. et al. Int. J. Bioi. Macromol. 2017, 98, 810-819);

2. CSIR-CDRI scientists, through in depth pharmacokinetics and toxicity assessments, have demonstrated that pluronic F-127 stabilized docetaxel nanocrystals improve apoptosis by mitochondrial depolarization in breast cancer cells (Pawar, V.K. et al. J. Biomed. Nanotechnol. 2015, 11 , 1747-1763;

3. CSIR-CDRI scientists have demonstrated that the both in vitro and in vivo antiproliferative activity of paclitaxel (PTX) in breast cancer cells are significantly enhanced when P is incorporated in a vitamin E nanoemulsion (Pawar, V.K. et al. J. Control Release. 2014, 196, 295-306; IF= 7.4);

1.CSIR-NCL scientists have developed a bioactive thermoresponsivepolyblend nanofiber formulations for wound healing (Pawar, M.D. et al. Mater Sci. Eng C Mater Bioi Appl.2015, 48, 126-137; PCT patent filed) . 2. CSIR-NCL scientists have deciphered the underlying mechanism of silk fibroin-

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WP4 (CSIR­IGIB)

WP5 (CSIR­CLRI)

WP6 (CSIR­IITR)

WP7 (lACS, Kolkata)

Targeting cell surface olecules for nucleic acid delivery

Development of targeted delivery system with biocompatible polymers for the connective tissue disorders and wound healing therapeutics (VENTURE) Development of cationic polymer-based delivery systems

Development of novel carbon nanotubes as functional carriers for drug delivery

Sophorolipid gelation (Dubey, P. et al. Biomacromolecules2016, 17, 3318-3327; IF 4.5) Several arginine-rich peptides have been designed and checked for their DNA delivery efficiency in multiple cell lines with different density of cell surface glycosaminoglycans (unpublished results) Enzyme-loaded nanofibers for wound healing; c) Lanthanum nanoparticle­loaded scaffold for biomedical applications have been achieved (Unpublished results)

CSIR-IITR scientists have demonstrated that hyaluronic acid grafted PLGA copolymer nanoparticles enhance the targeted delivery of Bromelain in Ehrlich's Ascites carcinoma (Bhatnagar, P. et al. Eur. J. Pharm. Biopharm. 2016, 105,295-306); Biotinylated amino aid based dispersing agent has been prepared by the trifnctionalisation of a tyrosine core. The acid terminal was functionalised with a PEG unit, the amine was attached to a biotin residue and the phenolic OH was converted to the hydrophobic segment by introduction of a C16 long cahin. The biotinylated amino acid based delivery vector showed significant, biocompatibility, stability, loading efficiency and selectivity in delivering biomolecules like oligoneucleotides and drugs into biotin receptor over expressesd cancer cells like HepG2 and Hela cells. Receptor mediated target specific delivery was achieved. [J Mat. Chern B (2014), 2, 1160-1173] Tumor vasculature targeting RGDK­decorated SWNP containg encapsulated curcutnin has been developed. The SWCNT NP-system has shown mouse tumor selective bio-distribution profiles

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WP8 (liSe., Bangalore)

WP9 (KEM Hospital, Mumbai

Developing Stealth Gemini surfactant and cationic polymer-lipid conjugate as cancer targeted delivery system

Evaluation of preclinical efficacies of lead DDS for infectious disease

upon i.v. administration in melanoma bearing C57/BL6J mice (Manuscript communicated to ACS Appl Mater Interface Sci) Synthesis of two cholesterol based redox-active geminicatioinic lipids and their men-cationic structural analogs have been completed. The physico-chemical characterizations of the liposomes of these novel cationic lipids have been completed. In vitro & in vivo studies are still going on in collaboration with IICT

HPLC Analytical methods for two APis (doxorubicinand daunorubicin) have been developed andvalidated. Two ligands have been synthesized for hepatictargeting. Ligand 1 is a glycolipid and has beencharacterised by IR, NMR (H1 and C13) and DSC,while ligand 2, a lipid conjugate, has been evaluated by IR and DSC. Characterisation by Mass Spectrometry for both the ligands is in progress.

The first batch of delivery systems received from the PI's group was tested for their in vivo tumor selective biodistributions. But it did not show tumor selective bio­d istribution.

The progress in this component of the project has been poor due to PI leaving country (moved to US University). The infectious disease part is

Conventional liposomal formulations for now being encapsulation of chosen drugs have handled by been developed. CSIR-CDRI

(leishmaniasis group)

5. Outcomes and outputs from the projects:

a. Outcomes

1. Lead(s) identified: Number and names with TRL

Number of leads with in vivo efficacy only

i) A dendritic cell based genetic immunization using in vivo DC-targeted liposomal DNA vaccine carrier in combination with tumor vasculature targeted chemotherapy, for combating "Established Tumor". TRL: 4

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ii) A glucocorticoid receptor (GR)-targeted liposomal based platform technology for delivering multiple drugs/genes to aggressive and relapsing

cancer models.

TRL: 4

iii)Gold nanoparticles-based new formulations for the delivery of drugs and nucleic acids for HER2+ cancer therapy. TRL: 4

iv) Sophorolipid and silk sericin formulation (cream) for wound healing application . TRL: 4

2. Technology/Product developed: Number and nameswith TRL level of Development

3. Technology/Product transferred: Number and nameswith name of

licensee(s) 4. Facility created : Number and names with uniqueness & utility 5. Sponsored projects obtained (with details) 6. External Cash Flow (ECF) generated 7. Societal impact created

b. Outputs:

1. Periodic Reports Submitted 2. Publications

i. Total number: 113 [Total scientists= 25] ii. CumulativeiF (as per 2015 figures): 470.063 iii. Average IF: 4.1598 iv. Average IF/scientist: 18.802

3. Patents (or other forms of IP generated such as Copyrights, Designs etc.) i. Filed : 15 ii. Granted iii . Licensed

4. Human Resource generated i. PhD fellows (give PhDs completed & pursuing separately)

PhD completed/submitted: 21 [including 1 (1) PA, who has completed her PhD]

PhD pursuing: 18 [including seven (7) PAs pursuing PhD with registration with AcSIR/ other Universities and 2 of them are now pursuing PhD with their independent fellowships (CSIR-SRF, 2 Nos.)].

ii. Project fellows: 20 iii. Skills imparted

- Organic synthetic chemistry skill ; skills related to detailed characterization including spectroscopic and chromatographic techniques etc.

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s. No

1.

- In vitro cellular biology, molecular biology skills

- In vivo skill for the development of tumor models such

subcutaneous, orthotopic models of lung , liver, brain, skin

cancers; Skill for the development of in vivo wound healing

models; biodistribution , pharmacokinetic and toxicity profiling

5. External collaborations established

1) [Under WP 1] Research collaborations with Mayo Clinic, USA; King Saud

University (KSU) , Saudi Arabia; CNRS/Univ. of Orleans, France; IACS­

Kolkata; 2) [Under WP 1.5] University of Science and Technology of Chian

(USTC); Universidad Rey Juan Carlos, Madrid , Spain ; IIT-Guwahati ; Au-KBC

Research Centre and Anna University, Chennai ; Sunshine Hospital,

Secunderabad; NRI Medical College, Guntur; lACS Kolkata.

Lead/Technology/ Salient TRL of the Lead/ Technology /Product

IP Status of Lead/Technology /Product

Status of Product (listed Features of the above outcomes)

in Lead/Technology/ Product

[From WP 1.1: Dr. Application of this TRL: 4 A. Chaudhuri]:A combination The dendritic cell based approach has combination genetic eradicated is ready to be immunization using established tested in in vivo DC-targeted melanoma, liver & large liposomal DNA colon cancer in animals. vaccine carrier in mouse tumor Toxicological combination with model. No need of studies for tumor vasculature targeted chemotherapy, for combating "Established Tumor

ex-vivo engineering of dendritic cells. The combination approach has markedly enhanced overall survivability of orthotopic lung tumor, pancreatic tumor and glioblastoma bearing mouse

the lipids used in combination approach should be evaluated in large animals.

Patent filed

DPR/Commer cial Technical Reportof Lead/ Technology/ Product NO

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2.

3.

4.

[From WP 1.2, Dr. R. Banerjee]: A glucocorticoid receptor (GR)­targeted liposomal based platform technology for delivering multiple drugs/genes to cancer stem cells (CSC), aggressive and relapsing cancer models

[From WP 1.5, Dr. C. R. Patra]Gold nanoparticles­based new formulations for the delivery of drugs and nucleic acids for HER2+ cancer therapy

[From WP 3.3, Dr. B. Garnaik] Solvent free process for preparation of high molecular weight polylactidecatalyze d by a compound of a divalent metal and an acid resulting in corresponding stereo selective polylactide, Patent (W02013108271A1 ), 2013.

This platform technology induces drug-sensitivity in CSCs, which epitomizes drug resistance and through EMT provides excessive aggressiveness to tumor. This also leads to relapsing phenotype in cancer, for which no chemotherapy is currently established . The platform system can carry both therapeutically important drug(s) and genes simultaneously. A dual strategy that can carry therapeutic siRNA and doxorubicin and targets specifically to ovarian cancer model in nude mice

High Molecular Weight PLA First time synthesized at NCL Toxicology study using OECD 420 guideline

TRL: 4 The technology is ready to be tested in large animal­based therapeutic and toxicological studies

TRL: 4 Toxicological assessment in small and large animals and PK studies in small animals is ready to be tested TRL: 1

Patent filed ND

Patent filed ND

Patent filed ND

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5.

6.

7.

8.

9.

[From WP 3.4, Dr. a Prabhune] Sophorolipid and silk sericin formulation (cream) for wound healing application.

[From WP 3.5, Dr. G V N Rathna] Antimicrobial non­woven nanofiber mat for wound healing device

[From WP 5] Functionalized nanoparticles as MRI contrast agents

[From WP 5] Bioactive incorporated Wound dressing

[From WP 5] Anti cancer Peptides

Faster Wound TRL:4 Healing, Low cost, No Scar Within 10 days the wound healed completely and no scar was observed

1. Oil based Polyesteramides were synthesized and blended with biocompatible polymers.

2. Fabricated with antimicrobial drugs as wound healing materials

3. In vitro toxicity and antibacterial studies were done

Filed patent (lndian/PCT}

TRL: 2

F-IONPs with TRL: 2 good cyto-compatibility and high magnetic property can serve as a novel platform for enhanced MRI sensitivity and drug delivery Standardized TRL: 2 different bioactives incorporated wound dressings Designed tumor TRL: 1 homing peptides

Patent filed ND

Patent filed ND

ND

ND

ND

6. Sectoral Monitoring Committee Recommendations on Lead/Technology/Product (listed above):

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Criteria SMC Recommendations

Name of LeadfTechnology i. Dendritic cell based genetic immunization /Product ii. Glucocorticoid receptor targeted liposome

Is the Lead/ Technology/Product Both of above worth taking up further?

What is the further R&D efforts that need to be put by CSIRICSIR laboratories?

Global benchmarking of the LeadfTechnology/Product specifically wrt specifications and cost

Is the Lead/Technology/Product worthy of commissioning a DPR at this stage?

What are the likely resources and time duration required for taking forward the identified Lead/ Technology/Product to the desired TRL?

Potential stake holders who may be appropriate to partner technically as well as financially

Suggestion for plausible road map towards further development of lead/technology/product for achieving desired TRL

GO/NO GO further development GO (Conditional- See comments below)

Other suggestions, if any, related The science needs to be evaluated more critically by domain to benefits/usage/ expert groups. The translational potential of these leads commercialization needs to be critically evaluated

7. Identified "lessons to be learnt", especially from shortcomings/failures.

8. Comments on financial progress on the project • fund availability and utilization of fund in the project:

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FIMnelal progreee le eatlefaetory and the lnvestmente mMe are juetlfled.

9. Sectoral Monitoring Committee Recommendations on Facility creation or other outcomes:

10. Grading of Project Execution: Outstanding/Excellent/ Very Good/ Good/ Satisfactory/Un· satisfactory

Very Good

11. Additional comments, if any:

~ Dr. C.~

(Member)

~~;iDt«_ {Member)

~h,)h Dr. S. Srikanth (Member)

~~~ Director, CSIR-IICB {TF Chairman-Member)

Director, CSIR-IMTECH (TF Chairman-Member)

Dr. T.S. Balganesh (Member)

w·· Dr. Ashok Rattan {Member)

s~~ra (Member)

Dime£.~ (TF Chairman· Member)

Director, CSIR-IICT (TF Chairman - Member)

u~ Prof P. Kondaiah

(Member)

Dr. M.K. Gurjar (Member)

H~11~ Director, CSIR-CDRI {TF Chairman -Member)

RA:tvt~ Director, CSIR-IIIM {TF- Chairman-Member)

Director, CSIR-NCL {TF Chairman - Member)

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Director, CSIR· NEIST (TF Chairman ·Member)

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